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18 April 2022, Volume 54 Issue 2
  • Establishment of a prediction model for colorectal cancer immune cell infiltration based on the cancer genome atlas (TCGA) database
    DING Ting-ting,ZENG Chu-xiong,HU Li-na,YU Ming-hua
    2022, (2):  203-208.  doi: 10.19723/j.issn.1671-167X.2022.02.001    
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    Objective: To study the correlation between immune cell infiltration in colorectal cancer tissue and clinical prognosis and to explore the levels of some immune cell genes for predicting the prognosis of patients with glioma colorectal cancer. Methods: In this study, we extracted colorectal cancer data from the cancer genome atlas (TCGA). Based on a deconvolution algorithm (called CIBERSORT) and clinically annotated expression profiles, the analysis assessed the infiltration patterns of 22 immune cells in colorectal cancer tissue to determine the association between each cell type and survival. Differences in five-year survival rate effectively illustrate the clinical prognostic value of each immune cell proportion in colorectal cancer, using a bar graph, correlation-based heatmap to represent the proportion of immune cells in each colorectal cancer sample. Results: A total of 473 colorectal cancer tissues and 41 normal control tissues were extracted from the TCGA database, and the comparative analysis showed that there were differences in the proportion of various TIICs in colorectal cancer tissues, which could characterize individual differences and have prognostic value. Among the cell subsets studied, the proportions of memory B cells, plasma cells, CD4+ T cells, natural killer (NK) cells, M0 macrophages, M2 macrophages, and activated mast cells were significantly different between normal and cancer tissues. Resting NK cells, CD8+ T cells, and plasma cells were associated with T phase, activated dendritic cells were associated with N phase, and eosinophils, M1 macrophages, and activated mast cells were associated with M phase. Survival analysis showed that activated dendritic cells were positively associated with five-year survival rate in colorectal cancer patients. Naive CD4+ T cells were inversely associated with five-year survival rate. Conclusion: There are different degrees of immune cell infiltration in colorectal cancer tissues, and these differences may be important determinants of prognosis and treatment response. We conducted a new gene expression-based study of immune cell subtype levels and prognosis in colorectal cancer, which has potential clinical prognostic value in colorectal cancer patients.

    Effects of CACNA1H gene knockout on autistic-like behaviors and the morphology of hippocampal neurons in mice
    JIAO Cui,WANG Jian-mei,KUANG Hai-xia,WU Zhi-hong,LIU Tao
    2022, (2):  209-216.  doi: 10.19723/j.issn.1671-167X.2022.02.002    
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    Objective: To investigate the effects of CACNA1H gene knockout (KO) on autistic-like behaviors and the morphology of hippocampal neurons in mice. Methods: In the study, 25 CACNA1H KO mice of 3-4 weeks old and C57BL/6 background were recruited as the experimental group, and 26 wild type (WT) mice of the same age and background were recruited as the control group. Three-chamber test and open field test were used to observe the social interaction, anxiety, and repetitive behaviors in mice. After that, their brain weight and size were measured, and the number of hippocampal neurons were observed by Nissl staining. Furthermore, the CACNA1H heterozygote mice were interbred with Thy1-GFP-O mice to generate CACNA1H -/--Thy1+(KO-GFP) and CACNA1H +/+-Thy1+ (WT-GFP) mice. The density and maturity of dendritic spines of hippocampal neurons were observed. Results: In the sociability test session of the three-chamber test, the KO mice spent more time in the chamber of the stranger mice than in the object one (F1,14=95.086, P<0.05; Post-Hoc: P<0.05), without any significant difference for the explored preference index between the two groups (t=1.044, P>0.05). However, in the social novelty recognition test session, no difference was observed between the time of the KO mice spend in the chamber of new stranger mice and the stranger one (F1,14=18.062, P<0.05; Post-Hoc: P>0.05), and the explored preference index of the KO mice was less than that of the control group (t=2.390, P<0.05). In the open field test, the KO mice spent less time in the center of the open field apparatus than the control group (t=2.503, P<0.05), but the self-grooming time was significantly increased compared with the control group (t=-2.299, P<0.05). Morphological results showed that the brain weight/body weight ratio (t=0.356, P>0.05) and brain size (t=-0.660, P>0.05) of the KO mice were not significantly different from those of the control group, but the number of neurons were significantly reduced in hippocampal dentate gyrus compared with the control group (t=2.323, P<0.05). Moreover, the density of dendritic spine of dentate gyrus neurons in the KO-GFP mice was significantly increased compared with the control group (t=-2.374, P<0.05), without any significant difference in spine maturity (t=-1.935, P>0.05). Conclusion: CACNA1H KO mice represent autistic-like behavior, which may be related to the decrease in the number of neurons and the increase in the density of dendritic spine in the dentate gyrus.

    Incidence and cause of abnormal cholesterol in children aged 2-18 years in a single center
    YAN Hui,PANG Lu,LI Xue-ying,YANG Wen-shuang,JIANG Shi-ju,LIU Ping,YAN Cun-ling
    2022, (2):  217-221.  doi: 10.19723/j.issn.1671-167X.2022.02.003    
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    Objective: To investigate the abnormality and distribution of plasma cholesterol levels in single-center hospitalized children. Methods: The blood lipid levels of children aged 2-18 years who had blood lipid test results in Peking University First Hospital from June 2016 to June 2019 were etrospectively analyzed. Cholesterol oxidase method was used for total cholesterol, and high-density lipoprotein cholesterol and low-density lipoprotein cholesterol were detected by clearance method. The counting data were compared with chi-square test. Results: The survey had involved 11 829 children (7 087 were boys and 4 742 were girls). 1 822 (15.4%) children were with elevated total cholesterol, 1 371 (11.6%) children with elevated low-density lipoprotein cholesterol, and 2 798 (23.7%) children with high-density lipoprotein cholesterol reduction. The total number of the children with abnormal cholesterol levels was 4 427 (37.4%). Among the 7 835 children who visited hospital due to the disease not commonly inducing dyslipidemia, 731 (9.3%) had elevated TC, 561 (7.2%) had elevated LDL-C, 1 886 (24.1%) had decreased HDL-C, and 2 576 (32.9%) had abnormal cholesterol levels. Among the children with different diseases, the difference in the incidence of abnormal cholesterol was statistically significant. The top three main groups of the children with increased total cholesterol and low-density lipoprotein cholesterol were “dyslipidemia”, “urinary tract disease”, and “nutritional disease”; The top three main groups of the children with reduced high-density lipoprotein cholesterol were “respiratory diseases”, “dyslipidemia”, “hematological diseases and malignant tumors”. Among the 1 257 blood li-pid test results sent by other departments, 300 cases had abnormal cholesterol levels (23.8%). Among them, there were 70 children with hypercholesterolemia (5.6%), 44 children with increased low-density lipoprotein cholesterol (3.5%), and 224 children with reduced high-density lipoprotein cholesterol (17.8%). There were 365 (4.6%) children with low-density lipoprotein cholesterol ≥140 mg/dL (3.6 mmol/L) who needed to further exclude familiar hypercholesterolemia among the children who visited hospitals due to the disease not commonly inducing dyslipidemia. Conclusion: Children in hospitals have a high incidence of cholesterol abnormalities. Doctors need to pay more attention to the cholesterol diagnosis and management regardless of the discipline, which not only helps to control secondary hypercholesterolemia, but also provides the possibility of detecting familial hypercholesterolemia in time.

    Clinical analysis of 30 cases of basal ganglia germinoma in children
    WANG Shu-lei,GAO Yang-xu,ZHANG Hong-wu,YANG Hai-bo,LI Hui,LI Yu,SHEN Li-xue,YAO Hong-xin
    2022, (2):  222-226.  doi: 10.19723/j.issn.1671-167X.2022.02.004    
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    Objective: To summarize and analyze the clinical characteristics of children with basal ganglia germinoma and to improve the level of early clinical diagnosis. Methods: The clinical data of children diagnosed with basal ganglia germinoma admitted to the Pediatric Surgery Ward of Peking University First Hospital from January 2013 to December 2020 were retrospectively analyzed,and descriptive statistics were used to analyze the clinical characteristics of children with basal ganglia germinoma. Results: A total of 30 patients were included in the study, 28 were male, 2 were female, the mean age at onset was (9.7±2.2) years, the median disease duration was 7 months, 27 had unilateral disease, and 3 had bilateral disease. The clinical manifestations were decreased limb muscle strength, cognitive function disorders, polydipsia, precocious puberty, intracranial hypertension, dysphonia and swallowing dysfunction. The serum and cerebrospinal fluid tumor marker alpha-fetoprotein (AFP) were normal in the 30 patients, and the serum and cerebrospinal fluid tumor marker β-human chorionic gonadotropin (β-HCG) were normal in 8 patients.The serum β-HCG was normal in 11 patients but the cerebrospinal fluid β-HCG was slightly elevated, and the serum and cerebrospinal fluid β-HCG were slightly elevated in 11 patients. A total of 33 lesions with irregular shapes were found by imaging examination, including 15 (45.5%) patchy lesions, 10 (30.3%) patchy lesions, and 8 (24.2%) round-like high-density lesions. Tumors showed obvious high-density shadows on computed tomography (CT) scan. Magnetic resonance imaging (MRI) scan of the tumors showed low or isointensity on T1WI and isointensity on T2WI, accompanied by mild peritumoral edema, hemispheric atrophy, cerebral peduncle atrophy, calcification, cystic degeneration,ventricular dilatation and wallerian degeneration. On contrast-enhanced scans, the tumor showed no enhancement or heterogeneous enhancement. Conclusion: The main age of onset of germ cell tumors in the basal ganglia in children is about 10 years old, and males are absolutely dominant. The clinical features and imaging manifestations have certain characteristics. With both combined, the early diagnosis of germ cell tumors in the basal ganglia can be improved.

    Short-term effects and seasonal variation of ozone on daily hospital outpatient visits for childhood asthma in Lanzhou
    ZHANG Hong,DONG Ji-yuan,WANG Jian-jun,FAN Lin-xia,QU Qiang,LIU Yang
    2022, (2):  227-235.  doi: 10.19723/j.issn.1671-167X.2022.02.005    
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    Objective: To analyze the effects of ozone (O3) concentrations measured with different approaches across different seasons on the total number of childhood asthma-related clinical visits, as well as the differentiation of such effects across different groups of patients. Methods: The outpatient data of three grade A tertiary hospitals in Lanzhou City spanning from 1 January 2014 to 31 December 2017, as well as air pollution and meteorological data during the same period were collected. Considering the nonlinear relationship between O3 concentrations and the total number of childhood asthma-related clinical visits and meteorological factors, a generalized additive temporal sequence model was employed to analyze the short-term association between changes in O3 concentrations and the total number of childhood asthma-related clinical visits. Taking into account of the variations in O3 concentrations within 1 day, this study adopted different measurement approaches to address the three types of O3 exposures, namely, the maximum 1 h daily concentration (O3max1h), the maximum 8 h daily concentration (O38h) and the mean 24 h daily concentration (O324h) as the short term exposure indicators to O3, followed by a model-based analysis. Results: The increase in short-term exposure levels to O3 in summer had a significant effect on the increase in the total number of childhood asthma-related clinical visits. With lag0 for the current day, every 10 μg/m3 increase in atmospheric concentration of O3max1h was associated with an increase in the total number of childhood asthma-related clinical visits by 3.351% (95%CI: 1.231%-5.516%); for every 10 μg/m3 increase in O38h concentration, the total number of childhood asthma-related clinical visits increased by 3.320% (95%CI: 0.197%-3.829%); for every 10 μg/m3increase in O324h concentration, the total number of childhood asthma-related clinical visits in summer increased by 6.600% (95%CI: 0.914%-12.607%); moreover, an increase in exposure to O3max1h also led to a significant rise in the total number of childhood asthma-related clinical visits among the males. Conclusion: The increase in short-term exposure levels to O3 in summer in Lanzhou City has a significant effect on the increase in the total number of childhood asthma-related clinical visits; O3max1h is more closely correlated with the increase in the total number of childhood asthma-related clinical visits.

    Effects of 275 nm and 310 nm ultraviolet irradiation on bone metabolism in ovariectomized osteoporotic rats
    HE Wei,YANG Si-wen,CHEN Juan,ZHU Xiao-jun,CHEN Zhi-zhong,MA Wen-jun
    2022, (2):  236-243.  doi: 10.19723/j.issn.1671-167X.2022.02.006    
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    Objective: To investigate the effect of 275 nm and 310 nm ultraviolet irradiation on ovariectomized rats’ bone metabolism. Methods: Twenty four 3-month-old female Sprague-Dawley (SD) rat were randomly divided into control group, sham operated group, 275 nm ultraviolet (UV) irradiation group and 310 nm UV irradiation group. Each group contained 6 rats. The rats in the two irradiation groups were treated with bilateral ovariectomy. The rats in sham operated group received sham operation (They were given the same back incision and a bit of par-ovarian fat were removed). Control group received no disposition. About 24 weeks after operation, all the rats received detailed bone mineral density (BMD) detection again. Detection regions include cervical vertebra, lumbar vertebra, proximal femur, mid femur and distal femur. Next, osteopenia rats in 275 nm irradiation group were UV irradiated 275 nm with fixed illumination intensity (15 μW/cm2) everyday for 16 weeks. The osteopenia rats in 310 nm irradiation group were UV irradiated 310 nm with fixed illumination intensity (15 μW/cm 2) everyday for 16 weeks. The backs of the rats were shaved regularly as irradiation area (6 cm×8 cm). After 16-week irradiation, all the rats’ BMD of cervical vertebra, lumbar vertebra, proximal femur, mid femur and distal femur were measured. At the end of the trial, all the rats’ blood specimens were obtained and serum 25(OH)D, procollagen type Ⅰ N-peptide (PINP) and osteocalcin (OC) were measured. Results: Compared with control group [(238.78±26.74) mg/cm3], the BMD of the whole body were significantly lower in 275 nm [(193.34±13.28) mg/cm3] and 310 nm [(191.19±18.48) mg/cm3] irradiation groups (P=0.002,P=0.001). There were no significant difference between sham operated group [(227.20±14.32) mg/cm3] and control group. After 16-week ultraviolet irradiation, the BMD of the whole body were significantly increased in 275 nm [(193.34±13.28) mg/cm3 vs. (221.68±25.52) mg/cm 3,P=0.005] and 310 nm groups [(191.19±18.48) mg/cm3 vs. (267.48±20.54) mg/cm 3,P<0.001] after corresponding irradiation. The BMD of the four body regions (lumbar vertebra, proximal femur, mid femur and distal femur) had significantly increased after irradiation in 275 nm irradiation group. For 310 nm irradiation group, the BMD in cervical vertebra, lumbar vertebra, proximal femur, mid femur and distal femur also had increased significantly after 310 nm ultraviolet irradiation. The concentration of serum 25(OH)D and OC was higher in 275 nm irradiation group than in control group [(46.78±5.59) μg/L vs. (21.32±6.65) μg/L, P=0.002;(2.05±0.53) U/L vs. (1.32±0.07) U/L,P=0.022]. Compared with the control, the concentration of serum 25(OH)D [(58.05±12.74) μg/L], OC [(2.04±0.53) U/L] and PINP [(176.16±24.18) U/L] was significantly higher (P<0.001, P=0.015, P=0.005) in 310 nm irradiation group. However, there were no significantly difference between sham operated group and the control. Conclusion: Both 275 nm and 310 nm ultraviolet could improve rats’ vitamin D synthesis. Both 275 nm and 310 nm ultraviolet could improve osteopenia rats’ bone condition. The irradiation of 310 nm might be more effective on bone condition improvement.

    Equol and its enantiomers inhibited urethane-induced lung cancer in mice
    YU Xue,ZOU Yong-qiu,WANG Ying,CHEN Ze-kun,MA De-fu
    2022, (2):  244-248.  doi: 10.19723/j.issn.1671-167X.2022.02.007    
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    Objective: To investigate the effects and mechanisms of equol and its enantiomers on urethane-induced lung cancer in mice. Methods: A total of 120 5-week-old male C57BL/6 mice were randomly divided into 8 groups: lung cancer tumor control group (CG), genistein control group (GCG), low dose racemic equol group (LEG), high dose racemic equol group (HEG), low dose R-equol group (LRE), high dose R-equol group (HRE), low dose S-equol group (LSE) and high dose S-equol group (HSE). Urethane was injected subcutaneously twice a week for 4 weeks to induce lung cancer and then the mice were fed for 4 months. The body weight and food intake of each group were measured and recorded weekly. After the mice were sacrificed, the blood, livers and lungs of the mice were collected. The incidence of lung cancer in each group was recorded. The concentration of serum superoxide dismutase (SOD), malondialdehyde (MDA) and 8-hydroxydeoxygunosine (8-OHdG) were detected by the corresponding kits. Western blotting was used to detect the expression of nuclear factor (erythroid-derived 2)-like 2 (Nrf2) in the livers. Between-group differences in body weight and food intake of the mice were compared using repeated measures ANOVA, and ANOVA for the differences between non-repeated measurements, with post hoc analysis using Tukey’s method if there were between-group differences. Comparisons of categorical data were performed by chi-square test, and if there were differences between the groups, the Bonferroni method was used for pairwise comparison. Results: A total of 49 in the 120 mice developed lung cancer. The overall incidence of lung cancer was 40.8%. Compared with the control group, the incidence of lung cancers in each experimental group was lower, and the difference was statistically significant. The incidence of lung cancer in the high-dose experimental group was significantly lower than that in the low-dose experimental group. However, the incidence of lung cancer was similar in the three equol groups and the genistein group at the same dose. Compared with the control group, the high-dose experimental group had higher serum SOD concentration, lower MDA and 8-OHdG concentrations, and the differences were statistically significant. Western blotting analysis showed that the expression levels of Nrf2 protein in the experimental groups were higher than those in the control group except the low-dose racemic equol group, and the Nrf2 protein expression level in the high-dose equol groups was higher than that in the low-dose equol groups. Conclusion: Racemic equol and its enantiomers mayinhibit lung carcinogenesis through antioxidant effects.

    Incidence and risk factors of ischemic stroke in patients with type 2 diabetes among urban workers in Beijing, China
    WU Jun-hui,WU Yi-qun,WU Yao,WANG Zi-jing,WU Tao,QIN Xue-ying,WANG Meng-ying,WANG Xiao-wen,WANG Jia-ting,HU Yong-hua
    2022, (2):  249-254.  doi: 10.19723/j.issn.1671-167X.2022.02.008    
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    Objective: To explore the incidence of ischemic stroke after the onset of type 2 diabetes, and further analyze the risk factors, so as to provide a basis for further research. Methods: The data were obtained from the database of the Beijing Urban Employee Basic Medical Insurance Database. The study used a prospective design to describe the incidence of ischemic stroke in patients with type 2 diabetes. In our study, these patients were followed up for seven years. Multivariate Logistic regression models were used to analyze the risk factors of ischemic stroke in patients with type 2 diabetes. Results: A total of 185 813 newly diagnosed type 2 diabetes patients were enrolled, with an average age of (58.5±13.2) years, and 49.0% of them were males. A total of 10 393 patients with newly diagnosed ischemic stroke occurred in 7 years, with a cumulative incidence of 5.6% and an incidence density of 8.1/1 000 person-years. Ischemic stroke occurred in all age groups in patients with type 2 diabetes. The cumulative incidence was 1.5% (95%CI: 1.3%-1.6%) in group ≤44 years old, 3.6% (95%CI: 3.4%-3.7%) in group 45-54 years old, 5.4% (95%CI: 5.2%-5.5%) in group 55-64 years old, and 9.2% (95%CI: 9.0%-9.4%) in group ≥65 years old, and the cumulative incidence increased with age (P<0.05). Cumulative incidence rate of the males (6.8%, 95%CI: 6.7%-7.0%) was higher than the females (4.4%, 95%CI: 4.3%-4.6%). Among the patients <80 years old, the cumulative incidence rate of the males was higher than that of the females in all the age groups. In the patients ≥80 years of age, the cumulative incidence was higher in the females (9.2%) than in the males (7.9%). Further analysis revealed that complications, such as coronary heart disease (OR=3.18, 95%CI: 2.72-3.72), heart failure (OR=1.53, 95%CI: 1.32-1.79) and kidney failure (OR=1.45, 95%CI: 1.20-1.75) were associated with ischemic stroke in the patients with type 2 diabetes. Conclusion: The incidence level of ischemic stroke in patients with type 2 diabetes is high. It is necessary to strengthen the management of risk factors in elderly patients, screen the complications of type 2 diabetes as early as possible, and take active preventive and control measures.

    A prospective cohort study of the influence of positive/negative effectivity on the mortality risk of the Chinese elderly
    LIU Jie,GUO Chao
    2022, (2):  255-260.  doi: 10.19723/j.issn.1671-167X.2022.02.009    
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    Objective: To explore the influence of positive/negative effectivity on the mortality risk of the Chinese elderly. Methods: Using the latest four surveys data of Chinese longitudinal healthy longevity survey in 2008, 2012, and 2014, as well as 2018 and selecting 10 993 elderly people who were aged 65 and above as research objects, this paper compared the positive/negative effectivity status and differences of the elderly in different gender and age groups. And then this paper analyzed the effect of positive/negative effectivity on elderly people’s mortality risk by constructing Cox proportional hazards model. Results: Compared with the female elderly, the male elderly had lower negative effectivity and higher positive effectivity. With the increase of age, the negative effectivity of the elderly increased, while the positive effectivity decreased. The results showed that after adjusting for the basic demographic characteristics, health status and living habits of the respondents in the baseline survey, in the positive effectivity dimension, “clean preference” (HR=0.922, 95%CI:0.889-0.956), “autonomy” (HR=0.933, 95%CI: 0.914-0.952) and “sense of youth” (HR=0.948, 95%CI:0.927-0.969), had a protective effect on the mortality risk of the elderly, while in the dimension of “open-minded view”, there was no significant difference in the mortality risk of the elderly with different answers. In the negative effecti-vity dimension, “tension and fear” (HR=1.039, 95%CI: 1.014-1.065), “loneliness” (HR=1.053, 95%CI:1.029-1.079) and “uselessness” had a harmful effect on the mortality risk of the elderly. The more positive effectivity, the lower the mortality risk of the elderly (HR=0.967, 95%CI: 0.956-0.978), while the more negative effectivity, the higher the mortality risk of the elderly (HR=1.024, 95%CI:1.013-1.035). Conclusion: The negative effectivity of the elderly in all dimensions may increase the mortality risk of elderly people. We ought to dredge the negative emotions of the elderly in time. In the health promotion of the elderly, we should also pay attention to the encouragement of positive effectivity in all dimensions, which requires the joint efforts of families, communities and governments.

    Analysis on the relationship between urbanization and health behavior in China: An empirical research based on China Health and Retirement Longitudinal Study (CHARLS)
    HE Shan,JIAN Wei-yan
    2022, (2):  261-266.  doi: 10.19723/j.issn.1671-167X.2022.02.010    
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    Objective: To explore the relationship between urbanization and residents’ health behaviors in China, and to clarify the impact of urbanization on health behaviors. Methods: Based on China Health and Retirement Longitudinal Study (CHARLS),we established a panel data fixed effect model tracked in 2011, 2013 and 2015 to analyze the effect of urbanization level on smoking cessation, drinking, sleep quality and physical exercise behavior. The ratio of population density, gross domestic product (GDP) per capita and output value of secondary and tertiary industries to GDP were used to represent the levels of urbanization. Results: From 2011 to 2015, China’s urbanization levels showed an increasing trend, which showed that the ratio of population density, GDP per capita and output value of secondary and tertiary industries in GDP increased year by year. Smoking cessation increased with the increase of GDP per capita and the proportion of the output value of secondary and tertiary industries. Compared with the low-level, the drinking rate in the regions with per capita GDP of more than 45 000 yuan increased by 2.49%, and the drinking rate in the regions with secondary and tertiary industries for 80%-85% decreased by 2.48%. However, there was no significant difference with population density. The sleep quality decreased with the increase of per capita GDP. In regions where per capita GDP was more than 93%, the sleep quality decreased by 3.71% compared with the low-level which was not significantly different from the ratio of population density and the output value of secondary and tertiary industries. For low contrast, the exercise rate in regions with the population density of 400-600 people/km2and over 800 people/km2 was reduced by 5.2% and 7.7% respectively. The exercise rate in regions with per capita GDP of 25 000-35 000 yuan and over 45 000 yuan was reduced by 3.26% and 3.73% respectively. The exercise rate in regions with secondary and tertiary industries accounting for more than 93% of GDP was 10.68% lower than that of the low-level regions. Conclusion: Different dimensions of urbanization have different impacts on different health behaviors. The smoking cessation rate increases with the increase of urbanization level, which is related to the proportion of per capita GDP and the output value of secondary and tertiary industries. The exercise rate, related to the three dimensions, decreases with the increase of urbanization. Sleep quality is more closely related to per capita GDP, and the probability of good sleep quality decreases with the increase of urbanization level. However, there is no obvious trend between drinking rate and urbanization level, which needs further study.

    Consistency analysis and clinical guiding significance of cytomegalovirus nucleic acid and antibody detections in patients with different clinical characteristics
    DAI Ju-hua,SUN Xin-ping,ZHANG Jie,SHI Lian-jie
    2022, (2):  267-271.  doi: 10.19723/j.issn.1671-167X.2022.02.011    
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    Objective: To investigate the consistency of cytomegalovirus deoxyribo nucleic acid (CMV-DNA) and immunoglobulin M (IgM) antibody detections in patients with different clinical characteristics and their guiding value for clinical practice. Methods: From December 2014 to November 2019, a total of 507 patients who were detected with both CMV-IgM and CMV-DNA were collected in Peking University International Hospital. Their general information, such as gender, age and clinical data, including the patient’s diagnosis, medication, and outcome were also collected. The groups were stratified according to whether CMV-DNA was negative or positive, CMV-IgM was negative or positive, age, gender, and whether they received immunosuppressive therapy or not. The Pearson Chi-square test or Fisher’s exact test was used for comparison of the rates between the groups. P<0.05 means the difference is statisti-cally significant. Results: Of the 507 patients submitted for examination, 55 (10.85%) were positive for CMV-DNA, 74 (14.60%) were positive for CMV-IgM, and 20 (3.94%) were positive for both CMV-DNA and CMV-IgM. Of the 55 patients with CMV-DNA positive, 37 were male, accounting for 67.27%. In addition, 25 patients were older than 60 years, accounting for 45.45% and 33 patients received immunosuppressive therapy, accounting for 60%. The rates were higher than that of CMV-DNA negative group, 47.35% (P=0.005), 68.14% (P=0.043), 46.02% (P=0.050), respectively. Of the patients with both CMV-DNA and IgM positive, 45% received immunosuppressive threapy, which was lower than that of CMV-DNA positive but IgM negative patients (68.57%, P=0.086), and also lower than CMV-DNA negative but IgM positive patients (68.52%, P=0.064). In the patients with both CMV-DNA and IgM positive, 91.67% showed remission after receiving ganciclovir, whereas in the patients with CMV-DNA positive but IgM negative, the rate was only 60% (P=0.067). Conclusion: CMV-IgM antibody detection is affected by age, gender, and immune status. It is not recommended to use CMV-IgM alone to determine CMV infection in patients with immunosuppressive status and those older than 60 years. CMV-DNA and CMV-IgM combined detection may help to predict patients’ immune status and outcomes of antiviral therapy.

    Effects of hepatitis B virus on Th17, Treg and Th17/Treg ratio in different alanine aminetransferase stages
    GAO Peng,LUO Yan-ping,LI Jun-feng,CHEN lin,MAO Xiao-rong
    2022, (2):  272-277.  doi: 10.19723/j.issn.1671-167X.2022.02.012    
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    Objective: To evaluate the effects of hepatitis B virus (HBV) on helper T lymphocytes 17 (Th17), regulatory T lymphocyte (Treg) and Th17/Treg ratio in chronic hepatitis B patients in different alanine aminetransferase (ALT) stages. Methods: In the study, 336 chronic hepatitis B patients in the first hospital of Lanzhou University were analyzed. The hepatitis B antigen antibody parameters were measured by chemiluminescence immunoassay analyzer, the liver function parameters were measured by automatic biochemical analyzer, the HBV loads were measured by quantitative PCR, Th17, Treg and Th17/Treg ratios were detected by flow cytometry. Among them, 111 cases (ALT<40 U/L) of ALT were normal hepatitis B, 108 cases of chronic hepatitis B with ALT above normal upper limit and <2 times higher (40 U/L≤ALT<80 U/L), and 117 cases of chronic hepatitis B with ALT above 2 times normal upper limit (80 U/L≤ALT). According to the viral load, they were divided into low replication group with HBV DNA < 4.0 lg copies/mL, medium replication group with 4.0 lg copies/mL≤HBV DNA<6.0 lg copies/mL and high replication group with HBV DNA ≥ 6.0 lg copies / mL. Dunnett T3 variance analysis were used to analyze the effects of HBV on Th17, Treg and Th17/Treg ratio in the chronic hepatitis B patients in different ALT stages. The changes of virological and immunological indexes before and after treatment were observed for 24 weeks of antiviral therapy in the hepatitis B patients with ALT≥double upper limit of normal group. Results: In the ALT normal group, different virus load HBV had minor effects on Th17, Treg and Th17/Treg ratio. In the ALT≥2 times upper limit of normal group, with the virus load increased, Th17 (3.18%±0.79% in low replication group, 3.78%±0.92% in medium replication group and 4.57%±1.15% in high replication group), Treg cells (5.52%±1.58% in low replication group, 5.89%±1.84% in medium replication group and 6.37%±2.35% in high replication group) and their ratio Th17/Treg (0.57±0.25 in low replication group, 0.65±0.29 in medium replication group and 0.73±0.36 in high replication group) were significantly increased (P<0.05). After entecavir treatment 24 weeks, the patient’s HBV-DNA decreased significantly, Th17 (3.89%±1.02% vs. 2.06%±0.46%), Treg (6.02%±2.03% vs. 5.06%±1.25%), Th17/Treg ratio (0.65±0.28 vs. 0.41±0.14) decreased significantly (P<0.05). Conclusion: Investigation on the effects of HBV on Th17 and Treg cells and their ratios in different ALT states can clarify the effects of HBV on the body from the immunological perspective and can further understand the ALT grouping for antiviral treatment theoretical significance, which is helpful for clinical treatment.

    Validation of the Pollard’s classification criteria (2010) for rheumatoid arthritis patients with fibromyalgia
    GAO Chao,CHEN Li-hong,WANG Li,YAO Hong,HUANG Xiao-wei,JIA Yu-bo,LIU Tian
    2022, (2):  278-282.  doi: 10.19723/j.issn.1671-167X.2022.02.013    
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    Objective: To evaluate the sensitivity and specificity of Pollard’s classification criteria(2010) for the diagnosis of rheumatoid arthritis (RA) patients withfibromyalgia (FM) in Chinese patients, and to assess the clinical features and psychological status of RA-FM patients in a real-world observational setting. Methods: Two hundred and two patients with rheumatoid arthritis were enrolled from the outpatients in Rheumatology and Immunology Department in Peking University People’s Hospital. All the patients were evaluated whether incorporating fibromyalgia translation occured using the 1990 American College of Rheumatolgy (ACR)-FM classification criteria. Forty two RA patients were concomitant with FM, while the other one hundred and sixty RA patients without FM were set as the control group. Results: There was no significant difference in general demography between the two groups (P>0.05). In this study, the Pollard’s classification criteria (2010) for RA-FM in Chinese patients had a high sensitivity of 95.2% and relatively low specificity of 52.6%. Compared with those patients without FM, RA patients with FM (RA-FM patients) had higher Disease Activity Scale in 28 joints (DAS-28) score (5.95 vs. 4.38, P=0.011) and much more 28-tender joint counts (TJC) (16.5 vs.4.5, P<0.001).RA-FM patients had worse Health Assessment Questionnaire (HAQ) score (1.24 vs. 0.66, P<0.001) and lower SF-36 (28.63 vs. 58.22, P<0.001). Fatigue was more common in RA-FM patients (88.1% vs. 50.6%,P<0.001) and the degree of fatigue was significantly increased in RA-FM patients (fatigue VAS 5.55 vs. 3.55, P<0.001). RA-FM patients also had higher anxiety (10 vs.4, P<0.001) and depression scores (12 vs.6, P<0.001). erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), morning stiffness time and 28-swollen joint counts (SJC) showed no difference between these two groups. Conclusion: The Pollard’s classification criteria (2010) for RA-FM are feasible in Chinese rheumatoid arthritis patients. The Pollard’s classification criteria is highly sensitive in clinical application, while the relativelylow specificity indicates that various factors need to be considered in combination. RA patients with FM result in higher disease activity,worse function aland psychological status. RA patients with FM also have poorer quality of life. DAS-28 scores may be overestimated in RA patients with FM. In a RA patient thatdoes not reach remission, the possibility of fibromyalgia should be con-sidered.

    Clinical efficacy of 585 nm Q-switched laser treatment on inflammatory lesion and postinflammatory erythema of acne vulgaris
    WANG Xiang-xi,LI Zhen-zhen,LAI Yan-yun,YANG Li,SHI Lin-li,ZHONG Shao-min,WU Yan
    2022, (2):  283-288.  doi: 10.19723/j.issn.1671-167X.2022.02.014    
    Abstract ( 51 )   HTML ( 1 )   PDF (1649KB) ( 15 )   Save
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    Objective: To evaluate the efficacy and safety of 585 nm Q-switched laser in the treatment of acne inflammatory lesions and postinflammatory erythema. Methods: A total of 25 patients with moderate facial acne, symmetrical distribution of inflammatory lesions and postinflammatory erythema on both sides of the face, were enrolled. Among the 25 patients, 22 patients completed all the treatment and evaluation, and 3 patients were lost to follow-up. 585 nm Q-switched laser was used on a randomly selected side of the face for three times of treatment at a 2 week interval. The evaluations were made before each treatment, 2 and 4 weeks after the last treatment, therefore the evaluation time points were before the treatment, weeks 2, 4, 6, and 8, respectively, for a total of 5 times. Acne severity was assessed using the investigator’s global assessment (IGA) score, and erythema severity was assessed using the investigator’s subjective erythema score and narrow-spectrum reflectance spectrophotometer at each follow-up. Results: After 3 times of treatment, there was statistically significant difference between the IGA score in week 8 and before treatment on both sides(Z=2.64, P<0.01; Z=2.67, P<0.01). There was no significant difference in IGA score between the treatment side and the control side before treatment and in week 8 (P=0.59, P=0.26). There was statistically significant difference between the investiga-tor’s subjective erythema score in week 8 and before treatment on the treatment side(Z=4.24, P<0.01), while no significant difference was showed on the control side(Z=1.73, P=0.08). In week 8, the investigator’s subjective erythema score of the treatment side was lower than that of the control side (Z=3.61, P<0.01). The erythema index of the treatment side was significantly decreased at 5 time points (P<0.01), and the index decreased significantly in week 8 compared with the index before treatment (P<0.01), while the erythema index of the control side was not significantly different at 5 time points. The treatment related adverse events included erythema and edema after treatment and pain during treatment, the severity was mild to moderate, which resolved spontaneously within 1 to 3 days. Nine patients were very satisfied with the treatment, 7 patients were satisfied, and 6 patients considered average. Conclusion: 585 nm Q-switched laser has some effect in the treatment of postinflammatory erythema, and it ensures good tolerance and safety. There was no statistically significant difference between the treatment side and the control side on the improvement of acne inflammatory lesions.

    Effect of dexamethasone combined with oxybuprocaine hydrochloride gel on prevention of postoperative sore throat after nasal endoscopy
    SHI Cheng-mei,WANG Xue-dong,LIU You-kun,DENG Ying,GUO Xiang-yang
    2022, (2):  289-293.  doi: 10.19723/j.issn.1671-167X.2022.02.015    
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    Objective: To explore the effectiveness and feasibility of dexamethasone combined with oxybuprocaine hydrochloride gel on the prevention of postoperative sore throat after nasal endoscopy. Methods: In the study, 60 patients with American Society of Anesthesiologist (ASA) physical statuses Ⅰ to Ⅱ, aged 18 to 72 years, scheduled for elective nasal endoscope surgery under general anesthesia requiring endotracheal intubation were randomly divided into dexamethasone combined with oxybuprocaine hydrochloride gel group (G group, n=30) and control group (C group, n=30). The patients in the G group received dexamethasone 0.1 mg/kg before induction and the oxybuprocaine gel was applied to the endotracheal catheter cuff and the front end within 15 cm. The patients in the C group received the same dose of saline and the saline was applied to the endotracheal catheter cuff and the front end within 15 cm. Then, all the patients in the two groups received the same induction and anesthesia maintainance. The operation time, anesthesia time, emergence time, extubation time and departure time were recorded. The intraoperative infusion volume, blood loss volume, propofol, remifentanil, rocuronium dosage were also recorded. The adverse reactions such as intraoperative hypotension, bradycardia and postoperative agitation were recorded. The postoperative sore throat score was recorded at the end of operation and 4 h, 8 h, 12 h, and 24 h after operation. Results: Compared with the C group, the emergence time [(8.4±3.9) min vs. (10.8±4.7) min], extubation time [(8.8±3.7) min vs. (11.9±4.8) min], and departure time [(20.0±5.3) min vs. (23.0±5.8) min] were significantly shorter, and the propofol dosage [(11.8±1.8) mg/kg vs. (15.9±4.6) mg/kg], remifentanil dosage [(10.9±4.7) μg/kg vs. (14.1±3.6) μg/kg] were significantly less in the G group, and there was no difference of rocuronium dosage in the two groups. Compared with the C group the incidence of intraoperative hypotension [10%(3/30) vs. 30%(9/30)], bradycardia [16.7%(5/30) vs. 20%(6/30)] and postoperative agitation [6.7%(2/30) vs. 23.3%(7/30)] were significantly lower in the C group. The postoperative sore throat score at the end of operation, 4 h, 8 h, 12 h and 24 h after operation in the G group were significantly lower than in the C group respectively [0 (0, 1) vs. 1 (1, 2), 0 (0, 0) vs. 1 (1, 2), 0 (0, 0) vs. 1 (1, 2), 0 (0, 0) vs. 1 (0.75, 1), 0 (0, 0) vs. 1 (0, 1)]. Conclusion: Dexamethasone combined with oxybuprocaine hydrochloride gel was effective and feasible on the prevention of postoperative sore throat after nasal endoscopy.

    Testosterone levels in patients with varicocele and azoospermia
    PENG Jing,FANG Dong,ZHANG Zhi-chao,GAO Bing,YUAN Yi-ming,TANG Yuan,SONG Wei-dong,CUI Wan-shou
    2022, (2):  294-298.  doi: 10.19723/j.issn.1671-167X.2022.02.016    
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    Objective: Androgen deficiency is common in aging males and may have unfavourable health consequences. Large-scale studies suggested low testosterone level might increse mortality and morbidity in ageing males. However, young men with low testosterone level might be neglected. Recent studies reported young men with infertility may have reduced testosterone level. To investigate the incidence of androgen deficiency in males with infertility and possible factors affecting the low testosterone level. Methods: Between January 2011 and December 2012, 407 men with infertility caused by varicocele (VC), obstructive azoospermia (OA) and nonobstructive azoospermia (NOA) in our center were included. The number of men in each group of OA, NOA and VC was 141, 97 and 169, respectively. All the eligible patients underwent a serum testosterone assessment by a single morning blood draw (between 8:00 to noon) to test for concentration of the total testosterone. All serum samples were determined by radioimmunoassay in our andrology laboratory. Androgen deficiency was defined as having a total testosterone level less than 300 ng/dL. Results: The mean age was (30.4±5.8) years. The mean testosterone level was (4.18±1.64) ng/dL (range 0.30 to 11.32 ng/dL). The overall incidence of androgen deficiency was 26.5% (108/407). The incidences of androgen deficiency in NOA, OA and VC groups were 40.2% (39/97), 19.1% (27/141) and 24.9% (42/169), respectively, which were significantly higher in the NOA than in the VC and OA groups (P<0.001). The incidences had no difference between the VC and OA groups (P=0.229). Univariate analysis revealed the cause of infertility, FSH and the mean testis volume as possible affecting factors for androgen deficiency. However, on multivariate analysis the only cause of infertility was an independent predictor. The incidence of androgen deficiency was the highest in the NOA group [OR 0.492 (95% confidence interval 0.288-0.840)]. Conclusion: NOA and varicocele might be risk factors of androgen deficiency. Young men with NOA may have a higher possibility of low testosterone level. Testosterone level should be followed up after NOA and varicocele treatment. Androgen deficiency should be assessed in males with infertility in clinical practice.

    Relationship between recovery of urinary continence after laparoscopic radical prostatectomy and preoperative/postoperative membranous urethral length
    ZHANG Fan,CHEN Qu,HAO Yi-chang,YAN Ye,LIU Cheng,HUANG Yi,MA Lu-lin
    2022, (2):  299-303.  doi: 10.19723/j.issn.1671-167X.2022.02.017    
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    Objective: To evaluate the relationship between recovery of urinary continence after laparoscopic radical prostatectomy (LRP) and preoperative/postoperative membranous urethral length (MUL) on magnetic resonance imaging. Methods: We retrospectively analyzed 69 patients with pathologic confirmed prostate carcinoma who underwent laparoscopic radical prostatectomy. Preoperative MUL was defined as the distance from the apex of prostate to the level of the urethra at penile bulb on the coronal image. Postoperative MUL was defined as the distance from the bladder neck to the level of the urethra at the penile bulb on the coronal image. MUL-retained rate was defined as the percentage of postoperative MUL to preoperative MUL. All patients received extraperitoneal LRP. Patients reported freedom from using safety pad (0 pad/d) were defined as urinary continence. Multivariate Logistic regression analyses were used to identify independent predictors of early continence recovery after LRP. Kaplan-Meier analyses and log-rank test were used to compare time to continence recovery between the groups. Results: For all the 69 patients, the average age was (71.4±8.6) years. The prostate specific antigen before biopsy was (23.40±30.31) μg/L, and the mean preoperative prostatic volume by magnetic resonance imaging was (39.48±22.73) mL. The mean preoperative MUL was (13.0±3.3) mm, the mean postoperative MUL was (12.3±3.4) mm, and the mean MUL-retained rate was 93.9%±6.2%. The continence rate for all the patients after LRP was 57.9% and 97.1% in three months and one year, respectively. The patients achieving early continence recovery had significant smaller prostatic volume (P=0.028), longer preoperative MUL and postoperative MUL (P<0.001). Multivariate Logistic regression analyses revealed postoperative MUL (P<0.001) were predictors of continence recovery after LRP. Kaplan-Meier analyses and Log-rank test revealed that preoperative MUL (≥14 mm vs. <14 mm, P<0.001) and postoperative MUL (≥13 mm vs. <13 mm, P<0.001), MUL-retained rate (<94% vs. ≥94%, P<0.001) were all significantly associated with continence recovery. Conclusion: Post-operative MUL was independently predictors of early continence recovery after LRP. Preoperative MUL, postoperative MUL and MUL retained rate were significantly associated with recovery of urinary continence.

    Clinical outcomes following microsurgery and endovascular embolization in the management of spinal dural arteriovenous fistula: A meta-analysis study
    YUAN Chang-wei,WANG Ying-jin,ZHANG Shu-jie,SHEN Sheng-li,DUAN Hong-zhou
    2022, (2):  304-314.  doi: 10.19723/j.issn.1671-167X.2022.02.018    
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    Objective: To compare the clinical effect of microsurgery and endovascular embolization in the treatment of spinal dural arteriovenous fistula (SDAVF) by meta-analysis. Methods: A systematic review was performed to retrieve all relevant literature about surgical treatment or endovascular embolization of SDAVF up to December 2019 through PubMed, Embase, Web of Science, Cochrane Central Register of Controlled Trials Results, CNKI, Wanfang Data, and SinoMed. The Chinese and English key words included: “SDAVF”, “spinal dural arteriovenous fistula”, “spinal AVM”, “spinal vascular malformation and treatment”. The included studies were evaluated using the Newcastle-Ottawa scale. The early failure rate, long-term recurrence, neurological recovery, and complications were evaluated and the clinical effects of the two methods in the treatment of SDAVF were compared by using RevMan 5.3 software. And a further subgroup analysis of the therapeutic effect of endovascular embolization with different embolic agents was conducted. Results: A total of 46 studies involving 1 958 cases of SDAVF were included, in which 935 cases were treated by microsurgery and 1 023 cases were treated by endovascular embolization. The funnel plot demonstrated that there was no publication bias. The results of meta-analysis showed that the incidence of early surgical failure was lower than that of endovascular embolization (OR=0.20, 95%CI: 0.13-0.30, P<0.05), and the long-term recurrence was also lower than that of endovascular embolization (OR=0.36, 95%CI: 0.22-0.58, P<0.05). The improvement of neurological function in the surgical patients is significantly higher than that in the patients treated with endovascular embolization (OR=2.86, 95%CI: 1.36-5.99, P<0.05). There was no significant difference in the occurrence of complications in these two groups (OR=1.52, 95%CI: 0.88-2.64, P=0.14). In the cases of endovascular embolization, the risk of treatment failure or recurrence was higher with Onyx glue than with n-butyl 2-cyanoacrylate (NBCA), and the difference was statistically significant (OR=4.70, 95%CI: 1.55-14.28, P<0.05). Conclusion: Although the treatment of dural arteriovenous fistulas by intravascular embolization has been widely used, the clinical effect of microsurgery is still better than that of endovascular embolization. Large scale and high-quality randomized controlled trials are required to validate the efficacy and safety of endovascular treatment in SDAVF patients.

    Microscopic resection of lumbar intraspinal tumor through keyhole approach: A clinical study of 54 cases
    LIN Guo-zhong,MA Chang-cheng,WU Chao,SI Yu
    2022, (2):  315-319.  doi: 10.19723/j.issn.1671-167X.2022.02.019    
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    Objective: To explore the feasibility and key technology of microscopic resection of lumbar intraspinal tumor through microchannel keyhole approach. Methods: The clinical features, imaging characteristics and surgical methods of 54 cases of lumbar intraspinal tumor which were microscopically operated by microchannel from February 2017 to September 2019 were reviewed and analyzed. There were 8 cases of extradural tumor, 3 cases of extra-and intradural tumor and 43 cases of subdural extramedullary tumor (including 3 cases of ventral spinal tumor). The tumors were 0.5-3.0 cm in diameter. The clinical symptoms included 49 cases of pain in the corresponding innervation area, 5 cases of sensory disturbance (numbness) at or below the tumor segment, 7 cases of limb weakness and 2 cases of urination and defecation dysfunction. Results: In the study, 37 tumors were resected through hemilaminectomy, 14 tumors were resected through interlaminar fenestration, 3 tumors were resected through hemilaminectomy or interlaminar fenestration combined with facetectomy of medial 1/4 facet. All of the 54 tumors were totally resected. The operation time was 75-135 min, with an average of 93.3 min. The postoperative hospital stay was 4-7 days, with an average of 5.7 days. Postoperative pathology included 34 cases of schwannoma, 4 cases of meningioma, 9 cases of ependymoma, 1 case of enterogenous cyst, 5 cases of teratoma/epidermoid/dermoid cyst, and 1 case of paraganglioma. No infection or cerebrospinal fluid leakage was found after operation. No neurological dysfunction occurred except 1 case of urination dysfunction and 4 cases of limb numbness. The follow-up period ranged from 3 to 33 months with an average of 14.4 months. Five patients with new onset symptoms returned to normal. The pain symptoms of 49 patients were completely relieved; 4 of 5 patients with hypoesthesia recovered completely, the other 1 patient had residual mild hypoesthesia; 7 patients with limb weakness, and 2 patients with urination and defecation dysfunction recovered to normal. No spinal instability or deformity was found, and no recurrence or residual tumors were found. According to McCormick classification, they were of all grade Ⅰ. Conclusion: The lumbar intraspinal extramedullary tumors within two segments (including the ventral spinal tumors) can be totally resected at stage Ⅰ through microchannel keyhole approach with appropriate selection of the cases. Microchannel technique is beneficial to preserve the normal structure and muscle attachment of lumbar spine, and to maintain the integrity and stability of lumbar spine.

    Knockdown of long non-coding RNA MIR4697 host gene inhibits adipogenic differentiation in bone marrow mesenchymal stem cells
    SHUAI Ting,LIU Juan,GUO Yan-yan,JIN Chan-yuan
    2022, (2):  320-326.  doi: 10.19723/j.issn.1671-167X.2022.02.020    
    Abstract ( 326 )   HTML ( 1 )   PDF (2777KB) ( 16 )   Save
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    Objective: To preliminarily investigate the role of long non-coding RNA (lncRNA) MIR4697 host gene (MIR4697HG) in regulating the adipogenic differentiation of bone marrow mesenchymal stem cells (BMSCs). Methods: For adipogenic differentiation, BMSCs were induced in adipogenic media for 10 days. The mRNA expression levels of lncRNA MIR4697HG and adipogenic marker genes including peroxisome proliferator-activated receptor γ (PPARγ), CCAAT/enhanced binding protein α (CEBP/α) and adiponectin (ADIPQ) were detected by quantitative real-time polymerase chain reaction (qRT-PCR) at different time points (0, 1, 2, 3, 5, 7, 10 days). The MIR4697HG stable knockdown-BMSC cell line was generated by infection of MIR4697HG shRNA-containing lentiviruses. To avoid off-target effect, two target sequences (shMIR4697HG-1, shMIR4697HG-2) were designed. And then cells were induced to differentiate in adipogenic medium. Oil red O staining, Western blot and qRT-PCR were used to detect the effect of MIR4697HG knockdown on adipogenic differentiation of BMSCs. Results: The mRNA expression level of MIR4697HG was significantly increased during adipogenic differentiation (P<0.01), and adipogenic differentiation of BMSCs was evidenced by upregulated mRNA levels of specific adipogenesis-related genes including PPARγ, CEBP/α and ADIPQ. Observed by fluorescence microscopy, more than 90% transfected target cells expressed green fluorescent protein successfully after shMIR4697HG-1 group, shMIR4697HG-2 group and shNC group transfection for 72 h. And the transfection efficiency of MIR4697HG examined by qRT-PCR was above 60%. Then the BMSCs were treated with adipogenic media for 7 days and showed that the mRNA expression levels of adipogenesis-related genes including PPARγ, CEBP/α and ADIPQ were significantly decreased in the MIR4697HG knockdown group (P<0.01), while the expression levels of PPARγ and CEBP/α proteins were decreased remarkably as well (P<0.01). Consistently, MIR4697HG knockdown BMSCs formed less lipid droplets compared with the control BMSCs, which further demonstrated that MIR4697HG knockdown inhibited adipogenic differentiation of BMSCs. Conclusion: lncRNA MIR4697HG played a crucial role in regulating the adipogenic differentiation of BMSCs, and MIR4697HG knockdown significantly inhibited the adipogenic differentiation of BMSCs. These data may suggest that lncRNA MIR4697HG could serve as a therapeutic potential target for the aberrant adipogenic differentiation-associated disorders including osteoporosis.

    Initial establishment of digital reference standardized crown models of the primary teeth
    FENG Sha-wei,GUO Hui,WANG Yong,ZHAO Yi-jiao,LIU He
    2022, (2):  327-334.  doi: 10.19723/j.issn.1671-167X.2022.02.021    
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    Objective: To explore the construction process of the digital reference crown models, and to initially establish the digital reference crown models of the primary teeth to lay the foundation for the establishment of the standardized crown models and the future related applications of computer-aided design/computer-aided manufacture (CAD/CAM) technology to pediatric dentistry. Methods: This study randomly selected children who were caries free, aged from 4 to 5 years in several kindergartens of Haidian District of Beijing. Plaster dental models were made for the children after taking complete impressions. The digital dental models were reconstructed by using the three-dimensional (3D) dental model scanner. And then, Geomagic Studio, a 3D reverse engineering software, was employed to extract the single dental crown data, the mesiodistal and buccolingual diameters and the height of the crowns were measured. The object was reduced or enlarged by a numerical factor, and then the size of each dental crown was standardized. A total of 3-5 points features on the crown were created, and all the objects were aligned through the functions of feature-based alignment. Finally, through average-based object creation and smoothing, the digital models of reference crowns of the primary teeth were established. Results: A total of 40 plaster dental models from 16 boys and 26 girls were selected out for our further study. The digital dental models were reconstructed, and the mesiodistal and buccolingual diameters and the height of the crowns were measured by using reverse engineering technology. Comparing the results of using mesiodistal diameter, buccolingual diameter and height as the standards, we chose the mesiodistal diameters of crowns to do the standardization, and successfully established the digital reference models of 20 primary teeth crowns with detailed surface characteristics. Conclusion: In this study, the digital reference crown models of the primary teeth were established by reverse engineering technology, providing reference value for the standardized crown models and application for clinical practice, scientific research and teaching. Furthermore, this study also contributes to the extensive application of CAD/CAM technology in pediatric dentistry and the development of CAD/CAM dental systems with independent intellectual property rights.

    Clinicopathologic analysis of micro and mini parotid gland tumors
    LI Bing-yu,TANG Zu-nan,HU Lei-hao,ZHANG Wen-bo,YU Yao,YU Guang-yan,PENG Xin
    2022, (2):  335-339.  doi: 10.19723/j.issn.1671-167X.2022.02.022    
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    Objective: To investigate the clinicopathological characteristics of micro and mini parotid gland tumors and to provide reference for their clinical diagnosis and treatment. Methods: Patients with parotid gland tumors treated in the Department of Oral and Maxillofacial Surgery, Peking University School and Hospital of Stomatology from December 2012 to April 2020 were selected. Relevant clinical data of the patients with tumor diameter ≤20 mm detected by preoperative CT were collected to analyze the clinicopathological characteristics and prognosis of micro and mini parotid gland tumors. And the collected data were divided into two groups with diameter 11-20 mm and diameter ≤10 mm according to tumor diameter measured by preoperative CT. The clinicopathological differences between the two groups were statistically analyzed. Results: A total of 2 067 patients with primary epithelial parotid gland tumors were collected, and 685 patients with tumor diameter ≤20 mm were examined by CT, accounting for 33.1%. The ratio of male to female patients with micro and mini parotid gland tumors was 1 ∶1.93, the average age was (45.3±13.8) years (12-83 years), and the median course of disease was 12 months (1 week to 30 years). Among them, 635 cases (92.7%) were benign tumors, 50 cases (7.3%) were malignant tumors, and the ratio of benign to malignant was 12.7 ∶1. The most common benign tumor was pleomorphic adenoma, and the most common malignant tumor was mucoepidermoid carcinoma. The micro and mini parotid gland tumors were divided into 11-20 mm group (n=611) and ≤10 mm group (n=74), the clinical characteristics comparison of the two groups of gender ratio, average age, course of di-sease had no statistical difference (P>0.05). In the 11-20 mm diameter group, the percentage of benign and malignant tumor was 92.8% (567/611) and 7.2% (44/611) respectively, and the ratio of benign to malignant tumors was 12.9 ∶1. In the ≤10 mm diameter group, the percentage of benign and malignant tumor was 91.9% (68/74) and 8.1% (6/74) respectively, and the ratio of benign to malignant tumors was 11.3 ∶1. There was no significant difference between the two groups (P>0.05). Fifty patients with malignant tumor were followed up for the median follow-up period of 39.5 months (1-91 months). Local recurrence occurred in 2 patients with one death. The overall 2-year survival rate was 93.7% and the 5-year survival rate was 89.3%. Conclusion: The majority of micro and mini parotid gland tumors was benign lesion. There was a good prognosis for micro and mini parotid gland carcinoma. Early surgical treatment was recommended for micro and mini parotid gland tumors.

    Efficacy of vertical control by using mini-implant anchorage in maxillary posterior buccal area for Angle class Ⅱ extraction patients
    LIANG Wei,TANG Yao,HUANG Wen-bin,HAN Bing,LIN Jiu-xiang
    2022, (2):  340-345.  doi: 10.19723/j.issn.1671-167X.2022.02.023    
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    Objective: To investigate the efficacy of vertical control by using conventional mini-implant anchorage in maxillary posterior buccal area for Angle class Ⅱ extraction patients. Methods: Twenty-eight Angle class Ⅱ patients [9 males, 19 females, and age (22.6±2.8) years] were selected in this study. All of these patients were treated by using straight wire appliance with 4 premolars extraction and 2 mini-implant anchorage in maxillary posterior buccal area. In this study, the self-control method was used to measure and analyze the lateral radiographs taken before and after orthodontic treatment in each case, the main cephalometric analysis items were related to vertical changes. The digitized lateral radiographs were imported into Dolphin Imaging Software (version 11.5: Dolphin Imaging and Management Solutions, Chatsworth, California, USA), and marked points were traced. Each marked point was confirmed by two orthodontists. The same orthodontist performed measurement on the lateral radiographs over a period of time. All measurement items were required to be measured 3 times, and the average value was taken as the final measurement result. Results: Analysis of the cephalometric radiographs showed that, for vertical measurements after treatment, the differences of the following measurements were highly statistically significant (P<0.001): SN-MP decreased by (1.40±1.45) degrees on average, FMA decreased by (1.58±1.32) degrees on average, the back-to-front height ratio (S-Go/N-Me) decreased by 1.42%±1.43% on average, Y-axis angle decreased by (1.03±0.99) degrees on average, face angle increases by (1.37±1.05) degree on average; The following measurements were statistically significant (P<0.05): the average depression of the upper molars was (0.68±1.40) mm, and the average depression of the upper anterior teeth was (1.07±1.55) mm. The outcomes indicated that there was a certain degree of upper molar depression after the treatment, which produced a certain degree of counterclockwise rotation of the mandibular plane, resulting in a positive effect on the improvement of the profile. Conclusion: The conventional micro-implant anchorage in maxillary posterior buccal area has a certain vertical control ability, and can give rise to a certain counterclockwise rotation of the mandible, which would improve the profile of Angle Class Ⅱ patients.

    A cone-beam computed tomography evaluation of three-dimensional changes of circummaxillary sutures following maxillary protraction with alternate rapid palatal expansions and constrictions
    LIU Wei-tao,WANG Yi-ran,WANG Xue-dong,ZHOU Yan-heng
    2022, (2):  346-355.  doi: 10.19723/j.issn.1671-167X.2022.02.024    
    Abstract ( 50 )   HTML ( 4 )   PDF (1669KB) ( 21 )   Save
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    Objective: To assess three-dimensional (3D) changes of circummaxillary sutures following maxillary protraction with alternate rapid palatal expansions and constrictions (RPE/C) facemask protocol in maxillary retrusive children, and to investigate the relationship between the changes of circum-maxillary sutures and zygomaticomaxillary suture (ZMS) maturation, and to explore the factors of maxilla forward movement with RPE/C and facemask. Methods: In the study (clinical trial registration No: ChiCTR2000034909), 36 maxillary retrusive patients were recruited and block randomized to either the rapid palatal expansion (RPE) group or the RPE/C group. Patients aged 7 to 13 years, Class Ⅲ malocclusion, anterior crossbite, ANB less than 0°, Wits appraisal less than -2 mm, and A-Np less than 0 mm were included in the study. The RPE group received rapid palatal expansion, whereas the RPE/C group received alternate rapid palatal expansions and constrictions, and both with facemask protraction. Head orientations of cone-beam computed tomography (CBCT) images were implemented by Dolphin 11.7. 3D measurements of circummaxillary sutures on CBCT images were evaluated using Mimics 10.01 before (T0) and after treatment (T1). The changes were analyzed with independent t test, two-way ANOVA, Pearson correlation and regression analysis. Results: Two subjects in the RPE/C group were lost to follow-up. A total of 34 patients reached the completion criteria and were analyzed. Compared with the RPE group, sagittal changes of circummaxillary sutures were significantly increased in the RPE/C group with 1.21 mm advancement of zygomaticotemporal suture, 2.20 mm of ZMS, 1.43 mm of zygoma-ticofrontal suture (P<0.05, respectively). Except for the zygomaticotemporal suture, the rest forward sagittal changes of other circummaxillary sutures showed no major difference in terms of the ZMS maturation. The Spearman’s correlation in RPE/C indicated a strong positive correlation of sagittal changes between ZMS and point A (P<0.01) with a regression analysis R 2=42.5%. Conclusion: RPE/C might be more effective on the treatment of maxillary retrusive children. As one of the major mechanical loading sutures during orthopedic therapy, ZMS showed a strong positive correlation with point A on sagittal changes.

    Relationship between prognosis and different surgical treatments of zygomatic defects: A retrospective study
    LAN Lin,HE Yang,AN Jin-gang,ZHANG Yi
    2022, (2):  356-362.  doi: 10.19723/j.issn.1671-167X.2022.02.025    
    Abstract ( 49 )   HTML ( 6 )   PDF (4389KB) ( 16 )   Save
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    Objective: To evaluate the effect and summarize the characteristics of different treatment methods in repairing zygomatic defect. Methods: A total of 37 patients with zygomatic defect were reviewed in the Department of Oral and Maxillofacial Surgery, Peking University School and Hospital of Stomatology from August 2012 to August 2019. According to the anatomical scope of defect, the zygomatic defects were divided into four categories: Class 0, the defect did not involve changes in zygomatic structure or continuity, only deficiency in thickness or projection; Class Ⅰ, defect was located in the zygomatic body or involved only one process; Class Ⅱ, a single defect involved two processes; Class Ⅲa, referred to a single defect involving three processes and above; Class Ⅲb, referred to zygomatic defects associated with large maxillary defects. The etiology, defect time, defect size and characteristics of zygomatic defects, the repair and reconstruction methods, and postoperative complications were collected and analyzed. Postoperative computed tomography (CT) data were collected to evaluate the outcome of zygomatic protrusion. Chromatographic analysis was used to assess the postoperative stability. Results: Among the causes of defects, 25 cases (67.57%) were caused by trauma, and 11 cases (29.73%) were of surgical defects following tumor resection. We performed autologous bone grafts in 19 cases, 6 cases underwent vascularized tissue flap,5 cases underwent external implants alone, and 7 cases underwent vascularized tissue flap combined with external implants. After the recovery of the affected side, the average difference of the zygomatic projection between the navigation group and the non-navigation group was 0.45 mm (0.20-2.50 mm) and 1.60 mm (0.10-2.90 mm), with a significant difference (P=0.045). Two patients repaired with titanium mesh combined with anterolateral thigh flap had obvious deformation or fracture of titanium mesh; 2 patients with customized casting prosthesis had infection after surgery and fetched out the prosthesis finally. Conclusion: Autologous free grafts or alloplastic materials may be used in cases without significant structural changes. Pedicle skull flap or vascularized bone tissue flap is recommended for zygomatic bone defects with bone pillar destruction, chronic inflammation, oral and nasal communication or significant soft tissue insufficiency. Titanium mesh can be used to repair a large defect of zygomatic bone, and it is suggested to combine with vascularized bone flap transplantation.

    Clinical analysis of selective tracheostomy necessary for patients undergoing head and neck surgery with free flap reconstruction
    CAI Tian-yi,ZHANG Wen-bo,YU Yao,WANG Yang,MAO Chi,GUO Chuan-bin,YU Guang-yan,PENG Xin
    2022, (2):  363-368.  doi: 10.19723/j.issn.1671-167X.2022.02.026    
    Abstract ( 39 )   HTML ( 3 )   PDF (536KB) ( 15 )   Save
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    Objective: To discover the factors that may affect the use of selective tracheostomy among patients who have undergone head and neck surgeries with free flap reconstruction, so that the patients will not need tracheostomy nor receive the unnecessary treatment. Methods: Five hundred and thirty-three patients who had undergone head and neck surgery with free flap reconstruction operated by the same team of surgery at Department of Oral and Maxillofacial Surgery at Peking University School of Stomatology from 2015 to 2016 were reviewed. Three hundred and twenty-one (60.2%) of these patients underwent selective tracheostomy. All the patients’ demographic information, operation-related information, prior treatments, comorbidities and complications were recorded and analyzed. Results: The patients with defects of the tongue, mouth floor, oropharynx and bilateral mandible, who underwent neck dissection and with previous radiotherapy and smoking habit were more likely to get selective tracheostomy. Usage of bulky soft tissue flap might also add to the risk of airway obstruction and the need of selective tracheostomy, while other factors were not significantly related to the risk of postoperative airway obstruction and the patients could be kept safe without selective tracheostomy. Most cases without tracheostomy were kept safe except one case, while 8.39% of the patients with tracheostomy suffered from tracheostomy related complications, mainly pneumonia and hemorrhage of the tracheostomy wound, yet none led to serious consequences or even death. Conclusion: Selective tracheostomy is not necessary for patients who have undergone head and neck surgeries with free flap reconstruction except that there are defects at the tongue, oropharynx and mandible. Neck dissection, bulky soft tissue flap reconstruction, previous radiotherapy and smoking habit may also add to the risk of postoperative airway obstruction, while a favorable decision would involve a combination of all the above factors to assure the safety of the postoperative airway for the patients undergone head and neck surgeries with free flap reconstruction.

    Pituitary immune-related adverse events induced by programmed cell death protein 1 inhibitors in advanced lung cancer patients: A report of 3 cases
    GU Yang-chun,LIU Ying,XIE Chao,CAO Bao-shan
    2022, (2):  369-375.  doi: 10.19723/j.issn.1671-167X.2022.02.027    
    Abstract ( 348 )   HTML ( 25 )   PDF (1198KB) ( 87 )   Save
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    Programmed cell death protein 1 (PD-1) and its ligand 1 (PD-L1) have been widely used in lung cancer treatment, but their immune-related adverse events (irAEs) require intensive attention. Pituitary irAEs, including hypophysitis and hypopituitarism, are commonly induced by cytotoxic T lymphocyte antigen 4 inhibitors, but rarely by PD-1/PD-L1 inhibitors. Isolated adrenocorticotropic hormone(ACTH) deficiency (IAD) is a special subtype of pituitary irAEs, without any other pituitary hormone dysfunction, and with no enlargement of pituitary gland, either. Here, we described three patients with advanced lung cancer who developed IAD and other irAEs, after PD-1 inhibitor treatment. Case 1 was a 68-year-old male diagnosed with metastatic lung adenocarcinoma with high expression of PD-L1. He was treated with pembrolizumab monotherapy, and developed immune-related hepatitis, which was cured by high-dose methylprednisolone [0.5-1.0 mg/(kg·d)]. Eleven months later, the patient was diagnosed with primary gastric adenocarcinoma, and was treated with apatinib, in addition to pembrolizumab. After 17 doses of pembrolizumab, he developed severe nausea and asthenia, when methylprednisolone had been stopped for 10 months. His blood tests showed severe hyponatremia (121 mmol/L, reference 137-147 mmol/L, the same below), low levels of 8:00 a.m. cortisol (<1 μg/dL, reference 5-25 μg/dL, the same below) and ACTH (2.2 ng/L, reference 7.2-63.3 ng/L, the same below), and normal thyroid function, sex hormone and prolactin. Meanwhile, both his lung cancer and gastric cancer remained under good control. Case 2 was a 66-year-old male with metastatic lung adenocarcinoma, who was treated with a new PD-1 inhibitor, HX008, combined with chemotherapy (clinical trial number: CTR20202387). After 5 months of treatment (7 doses in total), his cancer exhibited partial response, but his nausea and vomiting suddenly exacerbated, with mild dyspnea and weakness in his lower limbs. His blood tests showed mild hyponatremia (135 mmol/L), low levels of 8:00 a.m. cortisol (4.3 μg/dL) and ACTH (1.5 ng/L), and normal thyroid function. His thoracic computed tomography revealed moderate immune-related pneumonitis simultaneously. Case 3 was a 63-year-old male with locally advanced squamous cell carcinoma. He was treated with first-line sintilimab combined with chemotherapy, which resulted in partial response, with mild immune-related rash. His cancer progressed after 5 cycles of treatment, and sintilimab was discontinued. Six months later, he developed asymptomatic hypoadrenocorticism, with low level of cortisol (1.5 μg/dL) at 8:00 a.m. and unresponsive ACTH (8.0 ng/L). After being rechallenged with another PD-1 inhibitor, teslelizumab, combined with chemotherapy, he had pulmonary infection, persistent low-grade fever, moderate asthenia, and severe hyponatremia (116 mmol/L). Meanwhile, his blood levels of 8:00 a.m. cortisol and ACTH were 3.1 μg/dL and 7.2 ng/L, respectively, with normal thyroid function, sex hormone and prolactin. All of the three patients had no headache or visual disturbance. Their pituitary magnetic resonance image showed no pituitary enlargement or stalk thickening, and no dynamic changes. They were all on hormone replacement therapy (HRT) with prednisone (2.5-5.0 mg/d), and resumed the PD-1 inhibitor treatment when symptoms relieved. In particular, Case 2 started with high-dose prednisone [1 mg/(kg·d)] because of simultaneous immune-related pneumonitis, and then tapered it to the HRT dose. His cortisol and ACTH levels returned to and stayed normal. However, the other two patients’ hypopituitarism did not recover. In summary, these cases demonstrated that the pituitary irAEs induced by PD-1 inhibitors could present as IAD, with a large time span of onset, non-specific clinical presentation, and different recovery patterns. Clinicians should monitor patients’ pituitary hormone regularly, during and at least 6 months after PD-1 inhibitor treatment, especially in patients with good oncological response to the treatment.

    Persistent and serious hyperkalemia after surgery of primary aldosteronism: A case report
    WANG Wei,CAI Lin,GAO Ying,GUO Xiao-hui,ZHANG Jun-qing
    2022, (2):  376-380.  doi: 10.19723/j.issn.1671-167X.2022.02.028    
    Abstract ( 101 )   HTML ( 2 )   PDF (754KB) ( 27 )   Save
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    Hyperkalemia was one of the complications after primary aldosteronism surgery. Hyperkalemia after primary aldosteronism surgery was uncommon in clinical practice, especially persistent and serious hyperkalemia was rare. This complication was not attached great importance in clinical work. A case about persistent and serious hyperkalemia after primary aldosteronism adrenal adenoma surgery was reported and the patient was followed-up for fourteen months in this study. This patient had a laparoscopic adrenalectomy due to primary aldosteronism. Hyperkalemia was detected one month after surgery of this patient, the highest level of plasma potassium was 7.0 mmol/L. The patient felt skin itchy, nausea, palpitation. Plasma aldosterone concentration fell to 2.12 ng/dL post-operation from 35.69 ng/dL pre-operation, zona glomerulosa insufficiency was confirmed by hormonal tests in this patient after surgery. And levels of 24 hours urinary potassium excretion declined. Decrease of aldosterone levels after surgery might be the cause of hyperkalemia. Hyperkalemia lasted for 14 months after surgery and kalemia-lowering drugs were needed. A systemic search with “primary aldosteronism”, “hyperkalemia”, “surgical treatment” was performed in PubMed and Wanfang Database for articles published between January 2009 and December 2019. Literature review indicated that the incidence of hyperkalemia after primary aldosteronism surgery was 6% to 29%. Most of them was mild to moderator hyperkalemia (plasma potassium 5.5 to 6.0 mmol/L) and transient. 19% to 33% in hyperkalemia patients was persistent hyperkalemia. Previous studies in the levels of plasma potassium reached the level as high as 7 mmol/L in our case were rare. Whether hypoaldosteronemia was the cause of hyperkalemia was not consistent in the published studies. Risk factors of hyperkalemia after primary aldosteronism surgery included kidney dysfunction, old age, long duration of hypertention. This paper aimed to improve doctors’ aweareness of hyperkalemia complication after primary aldosteronism surgery. Plasma potassium should be monitored closely after primary aldosteronism surgery, especially in the patients with risk factors. Some patients could have persistent and serious hyperkalemia, and need medicine treatment.

    Early loss of primary molar and permanent tooth germ caused by the use of devitalizer during primary molar root canal therapy: Two cases report
    TIAN Jing,QIN Man,CHEN Jie,XIA Bin
    2022, (2):  381-385.  doi: 10.19723/j.issn.1671-167X.2022.02.029    
    Abstract ( 59 )   HTML ( 8 )   PDF (3316KB) ( 48 )   Save
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    Devitalization has been widely used in the root canal therapy of primary and permanent teeth in China more than ten years ago. With the development of local anesthetic drugs and injection technologies, this treatment method with high potential risks has been gradually abandoned. However, a questionnaire survey targeted all the participants at the 2018 China Pediatric Dentistry Conference showed that the devitalizer utilization proportion was still as high as 38.1% (383/1 005), even though the ratio was much lower than 75.5% (105/139) in 2003. These doctors had pay more attention to tissue burn caused by devitalizer marginal leakage or direct leakage, and know how to identify and handle with devitalizer burn. Devitalizers were usually made of arsenic trioxide, metal arsenic or paraformaldehyde, which have cytotoxicity, allergenicity, mutagenicity, carcinogenicity, and teratogenic effects on animals. Marginal leakage of devitalizers have high risks of causing soft and hard tissue necrosis. Most of the dentists have an understanding of the potential damages of arsenic-containing devitalizers, so they will choose parafor-maldehyde with relatively less toxicity. Paraformaldehyde has a certain self-limitation, and there are few cases reported, so some dentists lack of vigilance. Paraformaldehyde can also causes tissue necrosis if leakage happens, and the treatment methods are similar to that of arsenic-containing devitalizers. When handling with devitalizers burn, the necrosed soft and hard tissue, for example gingiva, alveolar bone or teeth that cannot keep, must be completely removed until fresh blood appears, then rinse with large amount of saline and seal with iodoform gauze. This paper described two cases of devitalizer burn during the root canal treatment of primary molars, both of the doctors failed to identify the devitalizer burn symptoms in the early stage, thus didn’t do proper treatments immediately after burning. Resulting in the necrosis of large area of gingiva and alveolar bone, loss of primary molars and permanent tooth germs 1-2 months after devitalizer burn. This paper reported these two cases in detail in order to warn dentists the high risks of using any kind of devitalizers, help them learn how to identify and treat devitalizer burn, and remind them to stop using devitalizers as soon as possible.


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Sponsor: Peking University
Editor-in-Chief: ZHAN Qi-min
Executive Editor-in-Chief: ZENG Gui-fang
Editing and Publishing: Editorial Department of Journal of Peking University (Health Sciences)
ISSN: 1671-167X
CN: 11-4691/R