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18 April 2020, Volume 52 Issue 2
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  • Establishment and gene expression analysis of drug-resistant cell lines in hepatocellular carcinoma induced by sorafenib
    Bo MA,Zhi-hua TIAN,Li QU,Yue-xiang LIU,Hong ZHANG,Hui-rong DING
    2020, (2):  207-213.  doi: 10.19723/j.issn.1671-167X.2020.02.003    
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    Objective: To establish the drug-resistant cell lines of hepatocellular carcinoma (HCC) induced by sorafenib, and to screen out the high expression genes in drug-resistant cell lines of HCC induced by sorafenib, then to explore the genes related to sorafenib resistance in hepatocellular carcinoma.Methods: The human PLC and Huh7 cell lines were obtained, then the PLC and Huh7 drug-resistant cell lines were induced with sorafenib by using intermittent induction in vitro. CCK8 assay was used to detect the IC50 value of sorafenib for evaluation of drug sensitivity of hepatocellular carcinoma cell lines in PLC and Huh7. All the up regulated genes in PLC and Huh7 drug-resistant cell lines induced by sorafenib were screened out using high-throughput cDNA sequencing (RNA-Seq), Ualcan database was used to analyze the correlations between the up regulated genes in PLC and Huh7 drug-resistant cell lines induced and four clinical biological characteristics of hepatocellular carcinoma, including the gene expressions between normal samples and tumor samples, tumor stage, tumor grade, and patient overall survival, to find the genes that might be involved in the mechanism of sorafenib resistance of hepatocellular carcinoma.Results: All the up regulated genes detected by the using high-throughput cDNA sequencing (RNA-Seq) in PLC and Huh7 drug-resistant cell lines were further screened out by following conditions:(1) genes co-expressed in PLC and Huh7 drug-resistant cells induced by sorafenib, (2) the fold change was more than 4 times and the difference was statistically significant (P <0.05), the top 12 up regulated genes in PLC and Huh7 drug-resistant cell lines were found, which were TPSG1, CBX4, CLC, CLEC18C, LGI4, F2RL1, S100A6, HABP2, C15ORF48, ZG16, FOLH1, and EPCAM. Compared with the correlations between the twelve genes and the clinical biological characteristics by Ualcan database, the potentially significant gene CBX4 was screened out.Conclusion: The human PLC and Huh7 drug-resistant cell lines of hepatocellular carcinoma induced by sorafenib were successfully established. CBX4, the gene related to sorafenib resistance in hepatocellular carcinoma, was screened out by the high-throughput cDNA sequencing (RNA-Seq) and further analysis using Ualcan database, which is providing a powerful basis for further research on the mechanism of sorafenib resistance of hepatocellular carcinoma.

    Interaction between PSF and cytokeratin 18 mediates PSF relocation to cell membrane and maintains chemosensitivity of myeloid leukemia
    Si-mei REN,Lu-yao LONG,Cheng-shan XU
    2020, (2):  214-220.  doi: 10.19723/j.issn.1671-167X.2020.02.004    
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    Objective: To identify the chaperone of polypyrimidine tractor-binding protein-associated splicing factor (PSF) in myeloid leukemia cells, and to explore the mechanism and redistributive pattern to cell surface of PSF in chemo-sensitive HL60 cells and resistant HL60/DOX cells.Methods: The eukaryotic expression vector of PSF was transfected with liposomes transiently, then flow cytometry was used to detect the expression level of PSF on the cell surface 24 h, 48 h and 72 h after vector transfections. We constructed a chimeric expression vector, streptavidin binding peptide (SBP)-PSF, meanwhile this vector was transfected and made SBP-PSF fusion protein overexpress. In addition, we used streptavidin magnetic beads to precipitate the cellular chaperonin of PSF and then identified its chaperonin by mass spectrometry (MS). Lentiviral vectors containing cytokeratin18 (K18) interference sequences were transfected into 293T cells to prepare lentivirus. HL60 and HL60/DOX cells were infected with lentivirus to obtain stable interfering K18 cell lines. Next, flow cytometry was used to test the membrane relocation level of PSF. Together, these methods confirmed the similar or different mechanisms of the PSF redistributing to membrane synergistically mediated by K18 in HL60 and HL60/DOX cells.Results: The expression of membrane relocated PSF was detected every day for three days(at the end of 24 h,48 h and 72 h) after transient overexpression. The expressing rate of PSF on the cell surface was 22.4%±3.5%, 37.9%±6.0%, 58.3%±8.8%, respectively in sensitive HL60 cells, while that was 4.7%±0.5%, 3.9%±0.6%, 2.9%±0.6% , respectively in resistant HL60/DOX cells. The difference of expressing rate on each day was significant, P<0.01. We identified K18 detected by co-immunoprecipitation and mass spectrum assay which was the cellular chaperone of PSF. We found that K18 knockdown decreased the PSF expression level which redistributed on cell surface from 48.9%±5.4% to 6.2%±1.0% in sensitive HL60 cells, and from 9.11%±1.2% to 2.21%±0.51% in resistant HL60/DOX cells, respectively.Conclusion: K18 is the intracellular chaperonin of PSF. The interaction of PSF and K18 mediates its redistribution to cell membrane in sensitive cells. While in resistant cells, PSF failed to relocate at the cell surface and accumulated in cells, which mediated resistance to chemotherapeutics.

    Subcellular localization of GTPase of immunity-associated protein 2
    Hong-quan QIN,You ZHENG,Man-na WANG,Zheng-rong ZHANG,Zu-biao NIU,Li MA,Qiang SUN,Hong-yan Huang,Xiao-ning WANG
    2020, (2):  221-226.  doi: 10.19723/j.issn.1671-167X.2020.02.005    
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    Objective: To analyze the subcellular localization of GTPase of immunity-associated protein 2 (GIMAP2) for the further functional study.Methods: In the study,we first obtained the protein sequences of GTPase of immunity-associated protein 2 (GIMAP2) from National Center for Biotechnology Information (NCBI) database, and then performed a prediction analysis of its transmembrane structure, nuclear localization signal (NLS), nuclear export signal (NES) and subcellular localization through bioinformatics online tools. GIMAP2 gene amplified by PCR was inserted into the expression vector pQCXIP-mCherry-N1 and positive clones were selected by ampicillin resistance. After using methods to extract and purify, the sequenced recombinant plasmid pQCXIP-GIMAP2-mCherry, together with the retroviral packaging plasmids VSVG and Gag/pol, was transferred into HEK293FT cells by liposomes for virus packaging. The virus supernatant was collected 48 h after transfection and directly infected the human breast cancer cell line MDA-MB-436. Immunofluorescence staining was constructed to detect the localization of endogenous and exogenous GIMAP2 in MDA-MB-436 cells. Meanwhile, green fluorescent chemical dyes were used to label mitochondria, endoplasmic reticulum, and lipid droplets in living MDA-MB-436 cells stably expressing the GIMAP2-mCherry fusion protein. Images for the three dye-labeled organelles and GIMAP2-mCherry fusion protein were captured by super-resolution microscope N-SIM.Results: Bioinformatics analysis data showed that GIMAP2 protein composed of 337 amino acids might contain two transmembrane helix (TM) structures at the carboxyl terminus, of which TMs were estimated to contain 40-41 expected amino acids,followed by the residual protein structures toward the cytoplasmic side. NES was located at the 279-281 amino acids of the carboxyl terminus whereas NLS was not found. GIMAP2 might locate in the lumen of the endoplasmic reticulum. Sequencing results indicated that the expression vector pQCXIP-GIMAP2-mCherry was successfully constructed. Fluorescent staining confirmed that GIMAP2-mCherry fusion protein, co-localized well with endogenous GIMAP2, expressed successfully in the endoplasmic reticulum and on the surface of lipid droplets in MDA-MB-436 cells.Conclusion: GIMAP2 localizes in the endoplasmic reticulum and on the surface of LDs, suggesting potential involvement of GIMAP2 in lipid metabolism.

    Palmitoylome profiling indicates that androgens promote the palmitoylation of metabolism-related proteins in prostate cancer-derived LNCaP cells
    Wen-qing LI,Si-mei REN,Xing-bo LONG,Yu-qing TIAN
    2020, (2):  227-233.  doi: 10.19723/j.issn.1671-167X.2020.02.006    
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    Objective: To explore potential therapeutic targets other than androgen-deprivation treatment for prostate cancer by screening the proteins induced by androgen at palmitoylation modification level in LNCaP cells.Methods: The LNCaP cells were treated with androgen (Methyltrienolone, R1881, 5 nmol/L) or dimethyl sulfoxide (DMSO) for 24 h, and then labeled with alkynyl palmitic acid Alk-C16 (100 μmol/L). After that, the cells were collected, lysed, the total protein was extracted, agarose beads labeled with azide (1 mmol/L) were added, and the click-chemistry reaction was carried out at room temperature for 1 h. The covalent bond formed by click-chemistry reaction of azide and alkynyl group was used to enrich the palmitoylated proteins on agarose beads. Label-free quantitation (LFQ) was used to compare the protein palmitoylation level of R1881 treated and untreated cells to screen the proteins induced by androgen at palmitoylation modification level.Results: In this experiment, 907 potential palmitoylated proteins (mascot score>2, P<0.05) were identified, among which 430 proteins had LFQ values not zero at least twice. Among the 430 proteins, the palmitoylation levels of 92 candidates were increased by androgen treatment, and their LFQ values were significantly upregulated (>1.5-fold, P<0.05) in ≥2 samples of androgen-treated vs. untreated LNCaP cells. We also used the software of cytoscape to classify the 92 proteins, and found that the known functional proteins of them could be divided into three categories: metabolism related, protein folding related and translation initiation related. Among them, metabolism related proteins included lipid metabolism (6), glucose metabolism (7) and respiratory electron transport chain (8), and a small amount of amino acid metabolism (2) and other metabolism related proteins (2). Notably, the ratio of LFQ of cytochrome b-c1 complex subunit 2 (UQCRC2) was significantly (>3-fold, P<0.05) higher in androgen-treated cells compared with untreated cells, indicating that the palmitoylation level of UQCRC2 was enhanced by androgen most significantly than that of others. The second was long-chain acyl CoA dehydrogenase (ACADVL) related to lipid metabolism and glucose 6-phosphate dehydrogenase (PGD) related to glucose metabolism, but the LFQ ratio of them was less than 3-fold.Conclusion: The research on palmitoylation mechanism of metabolism, especially the proteins related to respiratory electron transport chain, will provide a new guidance for the diagnosis and treatment of prostate cancer and the development of targeted drugs.

    Clinical value of serum isoform [-2] proprostate-specific antigen and its derivatives in predicting aggressive prostate cancer
    Kui-xia SUN,Cun-ling YAN,Zhi-yan LI,Ping LIU,Wei ZHANG,Qun HE
    2020, (2):  234-239.  doi: 10.19723/j.issn.1671-167X.2020.02.007    
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    Objective: To explore the clinical value of serum isoform [-2] proprostate-specific antigen (p2PSA) and its derivatives %p2PSA and prostate health index (PHI) in predicting aggressive prostate cancer (PCa).Methods: The pre-operation serum and basic clinical data of 322 patients with PCa (including 143 patients diagnosed with PCa by transrectal ultrasound-guided prostate biopsy and 179 patients undergoing radical prostatectomy) in Peking University First Hospital were collected from August 2015 to May 2018. Serum total prostate-specific antigen (tPSA), free prostate antigen (fPSA) and fPSA/tPSA (f/t) and the p2PSA level of all these patients were measured on automatic immune analyzers DxI800, and then %p2PSA and PHI were calculated. The prostate pathologic result was considered as the gold standard to evaluate the Gleason score of the patients with PCa. Receiver operator curves (ROC) were used to assess the ability of p2PSA, %p2PSA and PHI to predict aggressive PCa (pathologic Gleason score≥7) compared with those traditional markers tPSA, fPSA and f/t.Results: Among these patients, the p2PSA, %p2PSA and PHI median levels were significantly higher in patients with pathologic Gleason score≥7 than those with Gleason score<7 (p2PSA: 30.22 ng/L vs. 18.33 ng/L; %p2PSA: 2.50 vs. 1.27; PHI:91.81 vs. 35.44;all P<0.01). The area under curve (AUC) of %p2PSA and PHI (0.770, 0.760) in predicting Gleason score≥7 were higher than those of the traditional indicators tPSA, fPSA and f/t (AUC were 0.648, 0.536 and 0.693, respectively). Among those patients diagnosed with PCa by transrectal ultrasound-guided prostate biopsy, the AUC of %p2PSA and PHI (AUC were 0.808 and 0.801, respectively) in predicting Gleason score≥7 were higher than those of the traditional indicators tPSA, fPSA and f/t (AUC were 0.729,0.655 and 0.665 respectively). Among those patients undergoing radical prostatectomy, PHI and %p2PSA also had the trend of higher predictive value than those of the traditional indicators. The AUC of %p2PSA and PHI were 0.798 and 0.744, respectively while the AUC of tPSA, fPSA and f/t were 0.625, 0.507 and 0.697, respectively.Conclusion: Compared with traditional markers tPSA, fPSA and f/t, %p2PSA and PHI had much higher predictive value for aggressive PCa, which may help clinicians to evaluate the therapeutic regime and make more appropriate management plan for the patients.

    Clinical evaluation of tumor-stroma ratio in pseudomyxoma peritonei from the appendix
    Ru MA,Xin-bao LI,Feng-cai YAN,Yu-lin LIN,Yan LI
    2020, (2):  240-246.  doi: 10.19723/j.issn.1671-167X.2020.02.008    
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    Objective: To evaluate the effect of tumor-stroma ratio (TSR) on disease progression and prognosis of pseudomyxoma peritonei (PMP) from the appendix.Methods: The study included 30 PMP patients with complete individual patient data, who underwent cytoreductive surgery (CRS) plus hyperthermic intraperitoneal chemotherapy (HIPEC) in Beijing Shijitan Hospital. Image-Pro Plus was used to quantitatively analyze the proportion of tumor and stromal areas in hematoxylin-eosin staining pathological images, from which TSR was derived. Correlation studies were conducted to evaluate the relationships between TSR and clinicopathological features, immunohistochemical characteristics, and prognosis of PMP.Results: Among 30 PMP patients, there were 16 males (53.3%) and 14 females (46.7%), with the mean age of (54.9±2.3) years. There were 15 cases (50.0%) of low-grade mucinous carcinoma peritonei (LMCP) and high-grade mucinous carcinoma peritonei (HMCP), respectively, with vascular tumor emboli occurring in 4 cases (13.3%), nerve invasion occurring in 3 cases (10.0%), and lymphatic metastasis occurring in 4 cases (13.3%). The median peritoneal cancer index (PCI) score was 36 (range: 3-39). The median TSR was 8% (range: 2%-24%), with TSR≤10% in 19 cases (63.3%) and TSR>10% in 11 cases (36.7%). Immunohistochemistry showed that 16 cases (53.3%) had Ki67 label index ≤ 50% and 14 cases (46.7%) > 50%. The mutation rate of p53 was 56.7% and the loss rate of MMR protein was 11.8%. In addition, the expression rates of MUC2, MUC5AC, CDX2, CK7, and CK20 were 66.7%, 100.0%, 82.6%, 56.0%, and 92.3%, respectively. There were significant correlations between TSR and histopathological types, nerve invasion, Ki67 label index, and p53 mutation (P<0.05 for all). At the end of the last follow-up, 21 patients (70.0%) died and 9 patients (30.0%) survived, including 6 patients survived with tumor. The median overall survival (OS) was 12.7 months (95%CI: 10.4-11.5 months), and the 1-, 2-, and 3-year survival rates were 60.5%, 32.3%, and 27.7%, respectively. The median OS was 19.4 months (95%CI: 3.0-35.9 months) in the TSR≤10% group, versus 12.6 months (95%CI: 0.7-24.5 months) in the TSR>10% group (χ 2=3.996, P=0.046).Conclusion: TSR is correlated with histopathological types, tumor proliferation, invasion behaviors and prognosis of PMP, thus could be a new prognostic indicator for PMP.

    Effect of Fei-Liu-Ping ointment combined with cyclophosphamide on lung cancer cell proliferation and acidic microenvironment
    Liang GENG,Jing LV,Jing FAN
    2020, (2):  247-253.  doi: 10.19723/j.issn.1671-167X.2020.02.009    
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    Objective: To observe the effects of Fei-Liu-Ping ointment and chemotherapy on mice with lung cancer, and to explore the inherent mechanism of action from the point of acidic microenvironment and apoptosis.Methods: First of all, the Lewis lung cancer transplanted mouse model was established. Therefore, they were treated by Fei-Liu-Ping ointment, cyclophosphamide, Fei-Liu-Ping ointment + cyclophosphamide and the saline as control. All the groups' tumor size, tumor growth rate and food consumption were recorded. The mice were sacrificed and the tumors were took out after 15 days' interventions. Then lactate relative concentrations were detected with lactate kits and the protein expressions of glucose transporter 4 (GLUT4), hexokinase 1 (HK1), glucose-regulated protein 78 (GRP78), carbo-nic anhydrase-Ⅸ (CA-Ⅸ) were detected through immunohistochemical staining. Flow cytometry was adopted to detect the percentage of apoptotic tumor cells and regulatory T cells (Treg), and the expression of hypoxia-inducible factor-1α (HIF-1α), Bcl-2, Bax, Caspase-3, interleukin-2 (IL-2) were tested through western blot.Results: The strongest inhibition effect and the lowest tumor growth rate was found in Fei-Liu-Ping ointment + cyclophosphamide group. There were significant differences between Fei-Liu-Ping ointment + cyclophosphamide group and saline group(P<0.05). And the highest food consumption was found in Fei-Liu-Ping ointment + cyclophosphamide group while there were no significant differences between Fei-Liu-Ping ointment + cyclophosphamide group and saline group (P>0.05). Further molecular biological detections found that the lowest lactate level and regulatory T cells ratio were found in Fei-Liu-Ping ointment + cyclophosphamide group and these expressions of GLUT4, HK1, GRP78, CA-Ⅸ were suppressed. There were significant differences between Fei-Liu-Ping ointment+cyclophosphamide group and saline group (P<0.05). In addition, the Fei-Liu-Ping ointment + cyclophosphamide group's cell apoptosis increased significantly compared with saline group and there were significant differences on expressions of HIF-1α, Bcl-2, Bax, Caspase-3, IL-2 for this group compared with saline group.Conclusion: Chemotherapy and Fei-Liu-Ping ointment had the synergistic effect on inhibiting tumor growth and improving the general conditions of tumor-bearing mice. The effect was partly owed to the improvement on tissue hypoxia, the inhibition of HIF-1α expression and the regulations on its downstream proteins, such as GLUT4, HK1, GRP78, and CA-Ⅸ. And then all these alterations led to the modulation tumor acidic microenvironment, the induced tumor cells apoptosis and suppression of T cells to regulatory T cells differentiation.

    Efficacy and safety of oral pyrotinib in HER2 positive metastatic breast cancer: real-world practice
    Guo-hong SONG,Hui-ping LI,Li-jun DI,Ying YAN,Han-fang JIANG,Ling XU,Dong-gui WAN,Ying LI,Mo-pei WANG,Yu XIAO,Ru-yan ZHANG,Ran RAN,Huan WANG
    2020, (2):  254-260.  doi: 10.19723/j.issn.1671-167X.2020.02.010    
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    Objective: Pyrotinib, a novel irreversible pan-ErbB receptor tyrosine kinase inhibitor, showed promising antitumor activity and acceptable tolerability in phase Ⅱ and phase Ⅲ randomized clinical trials. We assessed the activity and safety of oral pyrotinib for human epidermal growth factor receptor 2 (HER2) positive metastatic breast cancer patients in the real world.Methods: We retrospectively analyzed 72 HER2 positive metastatic breast cancer (MBC) patients who received oral pyrotinib based regimens at Beijing Cancer Hospital and other four hospitals (Peking University First Hospital, China-Japan Friendship Hospital, General Hospital of PLA, Peking University Third Hospital) from August 2018 to September 2019. Progression free survival (PFS), objective response rate (ORR), adverse events (AE) of pyrotinib were investigated.Results: Seventy-two patients with HER2 positive MBC were enrolled. The median age of the patients was 55 years (range: 32-79 years). Sixty-nine (95.8%) patients had received anti-HER2 treatment in the metastatic and/or (neo) adjuvant settings; 61 (84.7%) patients had received anti-HER2 treatments in the metastatic setting in terms of trastuzumab 56 (77.8%) patients, lapatinib 36 (50.0%) patients, and T-DM1 4 (5.6%) patients. Among these 72 patients who received oral pyrotinib based regimens, 62 (86.1%) patients received pyrotinib (±trastuzumab) in combination with chemotherapy, 6 (8.3%) patients received pyrotinib (±trastuzumab) in combination with endocrine therapy and 4 (5.6%) patients received pyrotinib (±trastuzumab). Sixty-five (90.3%) patients received 400 mg pyrotinib once daily as initial dose, and 7 (9.7%) patients received 320 mg. Objectiveresponse and safety to pyrotinib based therapy were evaluable in all the 72 patients. One (1.4%) patient achieved complete response (CR), 18 (25.0%) patients achieved partial response (PR), 41 (56.9%) patients had stable disease (SD), and 12 (16.7%) patients had progressive disease (PD). The ORR (CR+PR) was 26.4% and the median PFS was 7.6 months (95%CI: 5.5-9.7 months). Among the 36 patients with prior lapatinib therapy, the median PFS was 7.9 months (95%CI: 4.1-11.7 months). Among the 15 patients with brain metastasis, the median PFS was 6.0 months (95%CI: 2.2-9.8 months). The main toxicities related to pyrotinib were diarrhea in 57 (79.2%) cases, and 48 (66.7%) cases with grade 1-2 as well as 9 (12.5%) cases with grade 3.Conclusion:Pyrotinib based therapy is an effective treatment for patients with HER2 positive MBC, including patients with lapatinib treatment failure and brain metastasis, and the toxicities can be tolerated.

    Enteral nutrition support reduces toxicity of chemotherapy in patients with advanced or metastatic esophageal cancer
    Zhi-wei SUN,Jun JIA,Ying YANG,Chuan-ling LIU,Yan-jie XIAO,Jing YU,Xiao-dong ZHANG
    2020, (2):  261-268.  doi: 10.19723/j.issn.1671-167X.2020.02.011    
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    Objective: To assess the impact of enteral nutrition support on response and toxicity of the first-line chemotherapy in those patients with advanced or metastatic esophageal cancer.Methods: We collected the clinical data of 118 patients with unresectable advanced or metastatic esophageal cancer who received the first-line chemotherapy in our center from July 2014 to December 2016. All these 118 eso-phageal cancer patients were then divided into two groups: the nutrition group (received enteral nutrition support in addition to chemotherapy) and the control group (received chemotherapy only). Differences were analyzed before and after chemotherapy in each of the nutritional indicators including Karnofsky performance status (KPS), weight, body mass index (BMI), hemoglobin (Hb), number of lymphocytes (Lymph), total protein (TP), albumin (Alb), triglycerides (TG), total cholesterol (TC) in both groups. And differences of the efficacy and toxicities of the first-line chemotherapy between the two groups were also analyzed.Results: (1) Weight, BMI and Hb were all significantly decreased after chemotherapy in the control group (P<0.001), while there was no significant change of weight and BMI in the nutrition group, just with Hb decrease only. However, there was no significant change of all the other nutrition indicators after chemotherapy in both groups. (2) Compared with the control group, the nutrition group had significantly lower incidence of grade 3 to 4 hematologic toxicities after chemotherapy (15.4% vs. 42.1%, P=0.004). In addition, the incidence of grade 3 to 4 nonhematologic toxicities after chemotherapy was also lower in the nutrition group but without statistical significance (0 vs. 9.2%, P=0.123). Logistic regression model was then used for multivariate analysis to identify the factors that affected the toxicity of chemotherapy in these patients, and the results showed that nutrition therapy was an independent influencing factor of grade 3 or higher hematological toxicity after chemotherapy in the patients with esophageal cancer (P=0.008, RR=6.048, 95%CI: 1.589-23.027). (3) The response rate of chemotherapy between the control group and the nutrition group had not significant difference.Conclusion: Enteral nutrition support in addition to chemotherapy could improve nutrition status and reduce toxicity of chemotherapy in advanced or metastatic esophageal cancer patients.

    Effects of chlorogenic acid on glucose tolerance and its curve characteristics in high-fat diet-induced obesity rats
    Cheng-cheng GUO,Xiao-yuan ZHANG,Ying-xiang YU,Lan XIE,Cui-qing CHANG
    2020, (2):  269-274.  doi: 10.19723/j.issn.1671-167X.2020.02.012    
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    Objective: To observe the effect of chlorogenic acid (chlorogenic acid,CGA) on the glucose tolerance and its curve characteristics in high fat diet-induced obesity (diet-induced-obesity,DIO) rats, so as to provide scientific grounds for the development and utilization of CGA in early prevention and reversal of prediabetes.Methods: Eight of forty-six male Sprague-Dawley rats were randomly selected as the normal diet group (CON group), and the rest were fed with high-fat diet. After 4 weeks, 24 high-fat-induced obese rats were screened according to the criteria and then randomly divided into high fat diet group (HFD group), 50% CGA group and 98% CGA group. The CGA groups received intragastric administrations of 50% CGA and 98% CGA orally via a gavage needle once a day for 8 weeks, respectively, while the CON and HFD groups received a carrier solution (phosphate buffer saline, PBS). Their body weights were measured weekly and oral glucose tolerance test (OGTT) was performed every 4 weeks. Fasting insulin and insulin release were measured at the end of the study. Meanwhile, HOMA-IR and visceral fat percentage were calculated. Histopathological examination by hematoxylin and eosin staining method were evaluated in the pancreatic tissues.Results: Before the intervention of chlorogenic acid, blood glucose levels 120 min after glucose loading (P<0.05) and AUC-G (P<0.05) were increased in the HFD group when compared with the CON group, and the time to glucose peak was delayed after 4 weeks of chlorogenic acid intervention (P<0.05). After 8 weeks of intervention, the HOMA-IR index, the insulin levels at 0 min, 30 min, 60 min, and 120 min after glucose loading and AUC-I increased (P<0.05), and the histopathological examination showed obvious hyperplasia of pancreatic islets (P<0.05). Compared with the HFD group, there was no significant change in glucose tolerance and glucose peak time in 50%CGA and 98%CGA groups at the end of 4 weeks of intervention. How-ever, after 8 weeks of intervention, OGTT-60min,OGTT-120min blood glucose (P<0.05) were lower, HOMA-IR index and OGTT-0min, OGTT-120min serum insulin level decreased (P<0.05), the time to glucose peak shifted to an earlier timepoint (P<0.05), abnormal islet hyperplasia attenuated (P<0.05) in 50% CGA and 98% CGA groups. Also, the OGTT-30min serum insulin level was decreased (P<0.05) in 50% CGA group.Conclusion: Delay in time to glucose peak during the OGTT was one of the manifestations of impaired glucose tolerance in DIO rats, and 50% and 98% CGA could improve the glucose tolerance and delay in glucose peak time.

    Diagnosis and treatment of primary intraspinal abscess
    Chang-cheng MA,Zhen-yu WANG,Guo-zhong LIN
    2020, (2):  275-280.  doi: 10.19723/j.issn.1671-167X.2020.02.013    
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    Objective: To summarize the feature and treatment of the primary intraspinal abscess in order to improve the prognosis.Methods: In the study, 13 cases of primary intraspinal abscess of the recent 20 years were retrospectively analyzed. The history, etiology, pathogen, surgical methods and prognosis were summarized.Results: The course of the illness ranged from 7 days to 6 months. All the cases began with pain. Of the 13 patients, 10 had limb weakness. Five had a fever and 8 had increased white blood cells. As for distribution, 1 was in cervical vertebra, 1 in cervicothoracic junction, 1 in thoracic vertebra, 4 in thoracolumbar junction, and 6 in lumbosacral segment. The results of bacterial culture were positive in 4 cases, 3 cases were diagnosed as tuberculosis by pathological examination, and 1 case was recognized as infection of Brucella melitensis bacteria because of prior brucellosis. The pathogen of the remaining cases were unclear. All the cases received surgical treatment and pathology examination. The surgical aim was mainly removal of the lesion, decompression and drainage. Postoperatively anti-infection and glucocorticoid therapy were performed according to the pathogeny results and clinical experience. Incision abscesses were seen in 2 cases and reoperations including debridement and repair with transferred muscle flap were performed. Postoperative follow-up ranged from 6 months to 3 years (mean 1.8 years). One case suffered postoperative recurrence and the abscess spread along the vertebral canal. Reoperation was performed. Infections of all the cases were recovered completely and the nervous system signs were all improved in different degrees.Conclusion: The onset of primary intraspinal abscess is relatively urgent, mainly with pain. The lumbar and sacral vertebra is the predilection site. The bacterial culture is mostly negative. Early operation and use of sufficient amount of broad-spectrum antibiotic are recommended. If the incision abscess forms after the operation, it is advisable to transfer the muscle flap to repair the coloboma on the basis of debridement. In order to relieve edema of spinal cord and nerve root, the glucocorticoid can be used in the escort of antibiotics.

    Predictive values of the combination of Friedman stage and sleep oximetry done early after surgery in predicting the treatment outcomes of uvulopalatopharyngoplasty
    Bing-wan DONG,Jun-bo ZHANG,Shui-fang XIAO
    2020, (2):  281-284.  doi: 10.19723/j.issn.1671-167X.2020.02.014    
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    Objective: To explore the predictive values of the combination of preoperative data, including medical history and physical examination, and results of sleep oximetry performed early after the surgery (one week) in predicting the treatment outcomes of uvulopalatopharyngoplasty (UPPP) in adult patients with obstructive sleep apnea hypopnea syndrome (OSAHS).Methods: All the patients who were diagnosed with OSAHS, underwent UPPP treatment and had complete follow-up data in our institution between Jan 2013 and May 2016 were enrolled in our study. The pre-operative data of these patients, including age, physical examination, and polysomnography (PSG) data, were all exactly collected for analysis. All these patients had received the evaluation of subjective efficacy scale and the sleep oximetry test in early post-operative days (within one week). The long-term efficacy evaluations including PSG and subjective efficacy scale were all finished at least three months after the surgery.Results: In the research 61 patients were finally studied, including 25 responders (41.0%) and 36 non-responders (59.0%). Regression analysis revealed that the pre-operative Friedman stage and early post-operative oxygen desaturation index of ≥4% (ODI4) were found to be independently predictive parameters for the UPPP treatment outcomes (P<0.05). The ROC curve analysis was used to estimate the predictive values of ODI4 to the treatment outcomes, and it was calculated with an area under the curve of 0.822. Using the ODI4 of 15 as the cutoff value, the sensitivity and specificity of that calculated were up to 0.778 and 0.760 respectively. The response rate among the patients with an early post-operative ODI4 of <15 was 70.4%, which was significantly higher than that of the patients with an early post-operative ODI4 of ≥15, whose response rate was 17.6% (P<0.05). The similar results could also be obtained in the patients with pre-operative Friedman stage Ⅱ (87.5% vs.25.0%) and Friedman stage Ⅲ (33.3% vs. 6.7%).Conclusion: The combination of early post UPPP operation oximetry parameters, especially ODI4, and pre-operative Friedman stage could be used in better evaluating the potential surgical outcomes of UPPP.

    Analysis of anxiety, depression and related factors in patients with chronic lumbocrural pain before minimally invasive surgery
    Duan YI,Wei ZHU,Xiu-li MENG,Xiao-guang LIU,Shui-qing LI,Bin ZHU,Dong-lin JIA
    2020, (2):  285-289.  doi: 10.19723/j.issn.1671-167X.2020.02.015    
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    Objective: To investigate anxiety and/or depression status of patients with chronic lumbocrural pain, and to further analyze related risk factors of anxiety and/or depression .Methods: Retrospective analysis of the medical data of patients who suffered from chronic lumbocrural pain caused by lumbar disc herniation and/or lumbar spinal stenosis and received minimally invasive surgery from March 2018 to April 2018. General data (including age, gender, education levels, past history, sleep order and medical insurance), numeric rating scale(NRS), Japanese Orthopedic Association(JOA) back pain scale and hospital anxiety and depression scale(HADS) were collected for analysis. The basic demographic data and clinic data were analyzed, possible related risk factors associated were analyzed by univariate analysis, and multivariate Logistic regression analysis was further used to find the relative independent risk factors and included all the predictive variables with P values less than 0.05 as covariates.Results: A total of 91 patients met the inclusion criteria and finished this study, the mean HADS score for anxiety was 8.1±4.2, 48(52.7%) respondents were screened positive for anxiety, while the rest 43(47.3%) patients had negative anxiety state, the mean HDDS score for depression was 6.9±4.9, 38(41.8%) respondents were screened positive for depression, and the rest 53(58.2%) patients were not depressed, and 56(61.5%) patients experienced anxiety or depression. There were significant difference in sleep disorder, JOA score and leg NRS score between the patients with and without anxiety(P<0.05), and the significant differences were also found in age, sleep disorder and JOA score between the patients with and without depression(P<0.05), Logistic regression analysis further showed that the JOA score and sleep disorder were risk factors for anxiety, and the JOA score was risk factor for depression.Conclusion: Patients with chronic lumbocrural pain are often accompanied by anxiety and/or depression before minimally surgery, the low JOA score and sleep disturbance increased the risk of presenting anxiety, and the low JOA score increased the risk of developing depression. It is necessary to evaluate mental status and related risk factors before surgery.

    Analysis of risk factors for necrosis of femoral head after internal fixation surgery in young and mid-aged patients with femoral neck fracture
    Bing-chuan LIU,Chuan SUN,Yong XING,Fang ZHOU,Yun TIAN,Hong-quan JI,Zhi-shan ZHANG,Yan GUO,Yang LV,Zhong-wei YANG,Guo-jin HOU,Shan GAO
    2020, (2):  290-297.  doi: 10.19723/j.issn.1671-167X.2020.02.016    
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    Objective: To retrospectively analyze the risk factors of avascular necrosis of femoral head (ANFH) after internal fixation in young and mid-aged adults.Methods: From January 2007 to December 2017, femoral neck fracture patients (18-60 years old) treated by reduction and internal fixation were retrospectively studied in Peking University Third Hospital. We recorded their gender, age, body mass index (BMI), American Society of Anesthesiology (ASA) grade, reason of injury, fracture side, interval between injury and surgery, location of fracture line, Garden classification, Pauwels classification, reduction method (open or closed), internal fixation and reduction quality. The diagnosis of ANFH was confirmed based on X-ray and MRI images during the follow-up. The internal fixation method inclu-ded cannulated compression screw (CCS) or dynamic hip screw (DHS, with or without anti-rotation screw). χ 2 test and Logistic regression analysis were used to analyze the relationship between the various factors and postoperative ANFH. Results: A total of 113 patients were included in this study, including 63 males and 50 females with an average age of (43.17 ± 12.34) years. They were followed up by (25.08 ± 16.17) months. ASA grade included grade Ⅰ (21 cases), grade Ⅱ (55 cases) and grade Ⅲ (37 cases). The reasons of injury included low-energy trauma (76 cases) and high-energy (37 cases). The fracture line included subcapital type (37 cases), transverse type (74 cases) and basal type (2 cases). Garden classification included type Ⅰ (3 cases), type Ⅱ (46 cases), type Ⅲ (39 cases) and type Ⅳ (25 cases). Pauwels classification included type Ⅰ (21 cases), type Ⅱ (55 cases) and type Ⅲ (37 cases). Interval between injury and surgery was (3.88 ± 3.66) days, 108 patients and 5 patients performed closed and open reduction respectively. 63 patients performed CCS, and 50 patients performed DHS. The reduction quality included grade A (91 cases), grade B (18 cases) and grade C (4 cases). 18 patients developed ANFH after surgery, the incidence rate was 15.93%(18/113). The result of χ 2 test showed the reason of injury (OR=0.19, P < 0.01), Garden classification (OR=0.13, P < 0.01), Pauwels classification (OR=0.12, P = 0.02), internal fixation method (OR=3.29, P = 0.04) and reduction quality (OR=0.33, P < 0.01) were significantly associated with ANFH. These five factors were further included into the Logistic regression analysis, and its results showed that the reason of injury (OR=4.11, P = 0.03) and Garden classification (OR=4.85, P = 0.04) were statistically significant. Conclusion: The reason of injury, Garden classification, Pauwels classification, internal fixation and reduction quality may increase the risk of ANFH after surgery, and the reason of injury and Garden classification were much more significant.

    Application of multidisciplinary team (MDT) in the treatment of severe trauma
    Zhe DU,Wei HUANG,Zhi-wei WANG,Jing ZHOU,Jian XIONG,Ming LI,Peng ZHANG,Zhong-di LIU,Feng-xue ZHU,Chuan-lin WANG,Bao-guo JIANG,Tian-bing WANG
    2020, (2):  298-301.  doi: 10.19723/j.issn.1671-167X.2020.02.017    
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    Objective: To explore the effect of multi-disciplinary team (MDT) in general hospitals on severe trauma patients.Methods: This study reviewed the treatment of patients with severe trauma in trauma center of Peking University People's Hospital from March 2017 to April 2019. The baseline information: the patients' gender, age, injury mechanism, etc.; the start indicators: the Glasgow coma scale (GCS), trauma index (TI), injury severity score (ISS); the start related indicators: time for activation, time for MDT to arrive, time for CT scan, time for damage control surgery; patient treatment and prognosis: ICU (intensive care unit) length of stay, number of cured and discharged patients, number of dead cases, number of patients transferred to rehabilitation hospital, were all analyzed. It discussed the composition of MDT, the initiation scheme, the indicators of initiation of MDT for severe trauma, and analyzed the correlation between the application of MDT and the prognosis of patients.Results: From March 2017 to April 2019, 112 trauma patients were treated by MDT in Peking University People's Hospital. There were 69 males and 43 females. The minimum age was 15 years, the maximum age was 89 years, most of them were 36-55 years old. The main injury mechanism was traffic accident injury. The GCS, TI, ISS were 13.0±2.9, 13.0±2.8, and 21.5±11.9, respectively. It took 3.7±0.8 minutes to start the call, 6.1±0.9 minutes for MDT personnel to arrive at the emergency rescue area, 23.8±3.0 minutes for fast CT and 92.6±15.4 minutes for injury control operation. All the hospitalized patients were treated effectively. ICU (Intensive care unit) hospitalization time was 12.6±6.7 days. 55 discharged patients were cured, 5 died (1 died of hemorrhagic shock, 4 died of severe brain injury) and 52 transferred to rehabilitation hospital.Conclusion: The treatment of severe trauma patients by MDT in trauma center of general hospitals can greatly improve the ability and level of treatment of severe trauma patients, make up for the lack of treatment of severe trauma especially multiple trauma patients in large general hospitals, and improve the treatment effect of severe trauma patients. It provides a reference model for large general hospitals to treat patients with severe trauma and multiple trauma and for the construction of trauma centers.

    Analysis of special ehealth service for corona virus disease 2019 (COVID-19) pneumonia
    Wen FENG,Liang-nan ZHANG,Jing-yuan LI,Tian WEI,Ting-ting PENG,Dong-xu ZHANG,Zai-xin GUO,Wei-song WANG
    2020, (2):  302-307.  doi: 10.19723/j.issn.1671-167X.2020.02.018    
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    Objective: To analyze how governments, hospitals and information technology(IT) companies use Internet technology to provide online health services during the early stage of corona virus disease 2019 (COVID-19) epidemic in January 2020 in China, and then provide suggestions and coping strategies for the later stage and post-epidemic time.Methods: We searched for information on ehealth ser-vices related to the outbreak of COVID-19 in China. The sources of information were mainstream search engines such as Baidu and the popular interactive social platforms such as Webchat. The keywords were "Internet+pneumonia", "Internet clinic", "pneumonia online clinic" and so on. The time of information was from January 20 to February 3, 2020. The key information was extracted and encoded by two persons back-to-back. The coding information included: name of organization provider, launching time, location of provider, service items, user, health workers engaging in the service, and so on. The coded information was entered and analyzed with SPSS 24.0 and Excel.Results: There were totally 57 projects launched by local governments, hospitals and IT companies. Most of them were launched from January 24th to 27th, the hospital and government projects services regionally, especially in eastern provinces. In this study, 90.48% of the enterprises and 100.00% of the hospitals had online fever clinic and consultation services for COVID-19, 66.67% of the enterprises and 37.04% of the hospitals serviced derivative health problems. Only a few projects provided tele-medical consultation. There were individual projects that provided online health management for home quarantine people. Physicians were the main force of various projects. In some hospital projects, there were also nurses, pharmacists and professional technicians to provide featured consultation.Conclusion: Ehealth is useful and helpful for the health care system to rapidly cope with health demand during instantaneous and post epidemic time. Regional distribution of ehealth is unbalanced. There are institutional and technical feasibilities for the emergency application of Internet technology. However, community health centers seldom provide ehealth or connect with tertiary hospitals with Internet. Therefore, all kinds of providers within healthcare system should promote emergence ehealth. Tele-medical diagnosis and referral should be developed by local governments during COVID-19. The application of "Internet+medical treatment" in community medical institutions and synergy among various institutions should be promoted.

    Effects of temperature and relative humidity on the number of outpatients with chronic obstructive pulmonary disease and their interaction effect in Lanzhou, China
    Hai-rong BAO,Xiao-ju LIU,En-li TAN,Juan SHU,Ji-yuan DONG,Sheng LI
    2020, (2):  308-316.  doi: 10.19723/j.issn.1671-167X.2020.02.019    
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    Objective: To understand the relationships of daily average temperature and relative humi-dity with outpatient visit frequency of patients with chronic obstructive pulmonary disease, and whether temperature and relative humidity have a lag effect.Methods: The effects of daily average temperature, relative humidity, and their interaction in Lanzhou between January 2013 and December 2017 on the outpatient visit frequency of chronic obstructive pulmonary disease patients were analyzed using Poisson generalized linear regression model combined with distributed lag non-linear model.Results: There was a non-linear relationship between the daily average temperature and the outpatient visit frequency of chronic obstructive pulmonary disease patients. Between -12 ℃ and -8 ℃, the outpatient visit frequency increased gradually with the decrease of the daily average temperature, and the outpatient visit frequency of chronic obstructive pulmonary disease patients increased by 11.60% per 1 ℃ of temperature drop. The daily average relative humidity also presented a non-linear effect on the outpatient visit frequency chronic obstructive pulmonary disease patients. When the daily average relative humidity was in the range of 15%-28%, the outpatient visit frequency increased gradually with the decrease of relative humidity, and the outpatient visit frequency of COPD patients increased by 37.05% for every 1% decrease of relative humidity. A synergistic effect was found between air temperature and relative humidity on chronic obstructive pulmonary disease, that is, under different relative humidity, the effect of air temperature was different. When the daily average relative humidity ≤ 50% and the daily average temperature≤11 ℃, the effect of air temperature was the most obvious. For every 1 ℃ drop in temperature, the daily out-patient visit frequency of the whole population increased by 12.68% (5.62% in males and 7.56% in females; 5.24% in population < 65 years and 14.74% in population ≥ 65 years). When the daily average relative humidity > 50% and the daily average temperature ≤ 11 ℃, the daily outpatient visit frequency of the whole population increased by 9.00% for every 1 ℃ drop in temperature (< 65 years, 7.11%; ≥65 years, 10.93%). When the daily average temperature > 11 ℃, the temperature had no effect on the daily outpatient visit frequency of chronic obstructive pulmonary disease patients under different relative humidity.Conclusion: The presence of a certain extent of interaction is observed between daily average temperature and relative humidity. Low-temperature and dry environment (relative humidity ≤50%, temperature ≤11 ℃) as well as low-temperature and high-humidity environment (relative humidity > 50%, temperature ≤11 ℃) can both increase the risk of outpatient visit in chro-nic obstructive pulmonary disease patients.

    Trends of prevalence of excellent health status and physical fitness among Chinese Han students aged 13 to 18 years from 1985 to 2014
    Yi SONG,Dong-mei LUO,Pei-jin HU,Xiao-jin YAN,Jing-shu ZHANG,Yuan-ting LEI,Bing ZHANG,Jun MA
    2020, (2):  317-322.  doi: 10.19723/j.issn.1671-167X.2020.02.020    
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    Objective: To analyze the trends of prevalence of excellent health status and physical fitness among Chinese Han students aged 13 to 18 years from 1985 to 2014.Methods: In the study, 738 523 students aged 13 to 18 years were extracted from the 1985 to 2014 Chinese National Survey on Students' Constitution and Health. Height, weight, vital capacity and indicators of physical fitness were measured for each student. According to National Standards for Students' Physical Health (2014 Revision), students meeting an overall score ≥ 90.0 were considered to be of excellent health status and physical fitness. We used the Chi-square test to compare the differences in prevalence of excellent health status and physical fitness among different subgroups and draw maps of regional distribution of prevalence by using ArcGIS.Results: From 1985 to 2014, the average height, weight, and BMI for Chinese Han students aged 13 to 18 years increased consistently, while the average vital capacity and indicators of physical fitness fluctuated largely. The overall prevalence of excellent health status and physical fitness increased from 2.7% in 1985 to 4.4% in 1995, dropped consistently to 1.1% in 2005, rebounded 0.6 percentage points in 2010, and increased to 2.2% in 2014. In each survey year, the prevalence for the boys was always higher than for the girls (P<0.001), the prevalence for middle school students aged 13 to 15 years was always higher than for high school students aged 16-18 years (P<0.001), and the prevalence for students in eastern region was higher than in western and central regions (P<0.001). In 1985 and 1995, the prevalence in certain provinces in eastern and central regions was <1%. In 2005, almost half provinces (14/30) had a prevalence <1%. In 2014, provinces in eastern coastal areas had relatively high prevalence of excellent health status and physical fitness in students aged 13 to 18 years, while provinces in central and western regions had relatively low prevalence.Conclusion: A fluctuating trend of the prevalence of excellent health status and physical fitness has been observed among Chinese Han students aged 13 to 18 years during the past three decades. There is great difference between the current prevalence of excellent health status and physical fitness and the goal of Outline of the Healthy China 2030 Plan, thus, the effective interventions and strategies for promoting students' physical activity and physical fitness are urgently needed in China.

    Awareness and knowledge of oral cancer among 1 483 residents in Beijing
    Xing-hong ZHOU,Ying HUANG,Chao YUAN,Shu-guo ZHENG,Jie ZHANG,Jian-guo ZHANG
    2020, (2):  323-331.  doi: 10.19723/j.issn.1671-167X.2020.02.021    
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    Objective: To investigate the oral cancer awareness, its related knowledge among residents in Beijing.Methods: A questionnaire survey was conducted among Beijing residents (non-medical related personnel) on the knowledge of oral cancer and its prevention and treatment.Results: A total of 1 483 questionnaires were completed, including 663 males (44.6%) and 820 females (55.3%), ranging from 15 to 91 years. Lung cancer was the most mentioned as heard of (94.3%), followed by liver cancer (92.5%) and gastric cancer (92.4%), while oral cancer was the least heard of (47.7%). Family account (P=0.015), residence (P=0.028), income (P=0.024), frequency of brushing teeth (P<0.001), frequency of mouth self-examination (P<0.001) and chew betel nut chewing (P=0.015) were significantly associated with oral cancer awareness. Most respondents learned about oral cancer from TV programs (24.3%), phone news (22.0%), WeChat (11.3%) and chatting with friends or people around (14.6%). Only 34.9% knew that smoking was a risk factor of oral cancer, and this knowledge was associated with age (P=0.011), education level (P=0.007), frequency of brushing teeth (P<0.001), and frequency of mouth self-examination (P=0.002). Only 23.5% knew that drinking was a risk factor of oral cancer, and this knowledge was associated with education level (P=0.002), residence (P=0.022), frequency of brushing teeth (P=0.009), and frequency of mouth self-examination (P=0.005). Only 35.1% knew that betel nut chewing was a risk factor for oral cancer, and this know-ledge was associated with age (P=0.040), education level (P=0.002), family account (P=0.002), income (P=0.005), frequency of brushing teeth (P=0.001), frequency of mouth self-examination (P<0.001), and betel nut chewing (P=0.002). Only 35.7%, 21.6% and 16.9%, respectively, knew that long-term unhealed ulcers, white plaques and red plaques in the mouth were the early signs of oral cancer.Conclusion: This survey demonstrates a general lack of public awareness and knowledge about signs and symptoms of oral cancer, and its risk factors and treatment. Some residents have not formed good oral hygiene habits. Specific measures should be taken to improve the public awareness of oral cancer and related knowledge about prevention and treatment, such as the use of various new media for the public to promote their oral health knowledge, in order to reduce the incidence of oral cancer, improve early attendance rates and the patients' survival rates.

    Effect of vertical soft tissue thickness on clinical manifestation of peri-implant tissue in patients with periodontitis
    Zhong ZHANG,Huan-xin MENG,Jie HAN,Li ZHANG,Dong SHI
    2020, (2):  332-338.  doi: 10.19723/j.issn.1671-167X.2020.02.022    
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    Objective: To observe and investigate the effect of vertical soft tissue thickness on the peri-implant tissue condition and the prevalence of peri-implant disease in patients with history of periodontitis. Methods: Among 210 patients who showed initial interest of implant therapy, 92 patients were included in this study and received implant surgery during 2010 and 2015. Sixty-six patients with 66 implants finally came back for T2 evaluation. Prior to the implant therapy, all the patients had received periodontal treatment. During the implant placement surgery, the distance from palatal soft tissue edge to the alveolar crest, which was defined as vertical soft tissue thickness (VT), was measured after the buccal full thickness flap was elevated. According to the cut off point which was adopted from the operating characteristic curve, 66 implants within 66 patients were divided into two groups, which were called normal group (VT≤4.5 mm) and thick group (VT>4.5 mm), respectively. Information of the patient's general status, periodontal situation and implant information were recorded. After a follow-up period of 42.9 months, the parameters of peri-implant tissue and condition of peri-implant disease were recorded. Mann-Whitney U test as well as Chi-square test were used to compare the parameters between two groups. Moreover, Kaplan-Meier method was chosen to draw the event(peri-implantitis)-free survival curve. Results: The survival rate of the implants was 100%. At the end of the follow-up examination(T2), the parameters including max PDi, mean PDi, max BIi, mean BIi, mean MBL, MBL at distal side, MBL at mesial side, mean PLIi presented significantly higher values in thick group than in normal group (P < 0.05). Moreover, the prevalence of peri-implantitis and peri-implant disease(peri-implant mucositis & peri-implantitis) in thick group was respectively 34.8% and 73.9%, which was significantly higher than 2.3% and 48.8% respectively in normal group (P<0.05). The prevalence of peri-implant mucositis did not show significant difference in the two groups. In addition, Kaplan-Meier analysis showed that there was significant difference between the event-free survivals of the two groups. Conclusion: The vertical soft tissue thickness around implants in patients with periodontitis has a significant effect on the health of the peri-implant tissue. Excessive vertical soft tissue thickness may result in deeper peri-implant probing depth as well as more peri-implant marginal bone loss, and eventually increase the risk of peri-implant disease. The vertical remodeling of soft tissue may be a new direction to indicate the role of periodontitis in peri-implant tissue condition. Moreover, the biological mechanism of the association between soft tissue thickness and peri-implantitis risk as well as effective approaches to prevent the adverse effect of excessive soft tissue thickness on peri-implant tissue is necessary to be investigated.

    Gingival thickness assessment of gingival recession teeth
    Zi-yuan CHEN,Jin-sheng ZHONG,Xiang-ying OUYANG,Shuang-ying ZHOU,Ying XIE,Xin-zhe LOU
    2020, (2):  339-345.  doi: 10.19723/j.issn.1671-167X.2020.02.023    
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    Objective: To evaluate the gingival thickness and gingival biotype of gingival recession teeth of Chinese population.Methods: A total of 112 non-molar teeth with gingival recession in 34 patients were included. Direct measurement, cone-beam computerized tomography (CBCT) measurement and periodontal probe method were used to evaluate gingival thickness and biotype. Gingival thickness was measured at 2 mm apical to the gingival margin. Direct measurement was performed with a caliper of 0.01 mm resolution and anesthesia needles attached to silicone disk stops. Gingival biotype was assessed by sulcus probing, if the periodontal probe was visible through the gingival tissue, the gingival biotype was thin; If not visible, the gingival biotype was thick. The differences of gingival thickness among different gingival biotype, tooth site and gingival recession type were analyzed respectively. Besides, the results of CBCT measurement was analyzed compared with the direct measurement.Results: The average gingival thickness of non-molar recession teeth was (1.17±0.41) mm. The average gingival thickness of thick and thin biotype group were (1.38±0.4) mm and (0.97±0.30) mm, respectively, with statistically significant difference (P<0.001).The median of gingival thickness was 1.1 mm. Using 1.1 mm as the cut-off value of thick and thin gingival thickness group, the results matched well with the gingival biotype classification results by periodontal probe method (P=1.000). The average gingival thickness of maxillary teeth was significantly thicker than that of the mandibular teeth. They were (1.39±3.44) mm and (1.01±0.31) mm, respectively (P<0.001). The mean gingival thickness of MillerⅠ, Ⅱ and Ⅲ degree gingival recession teeth were (1.15±0.34) mm, (0.83±0.17) mm and (1.26±0.56) mm, respectively, without statistically significant difference (P=0.205). The gingival thickness measurement results between CBCT method and direct measurement were without statistically significant difference (P=0.206).Conclusion: In the non-molar gingival recession teeth, the cut-off value of gingival thickness to classify thick and thin biotype of Chinese population was 1.1 mm. The average gingival thickness of the maxillary teeth was significantly thicker than that of the mandibular teeth. Besides, CBCT measurement was an accuracy method for evaluating facial gingival thickness.

    Effect of concentrated growth factors combined with guided tissue regeneration in treatment of classⅡ furcation involvements of mandibular molars
    Fei LI,Jing QIAO,Jin-yu DUAN,Yong ZHANG,Xiu-jing WANG
    2020, (2):  346-352.  doi: 10.19723/j.issn.1671-167X.2020.02.024    
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    Objective: Tissues loss due to periodontal disease is typically treated by a variety of rege-nerative treatment modalities, including bone grafts, guided tissue regeneration (GTR) and growth factors, to reform the supporting tissues of teeth. Concentrated growth factors (CGF) are produced by centrifuging blood samples at alternating and controlled speeds using a special centrifuge. The purpose of this study was to evaluate whether GTR could improve the effect of CGF combined with bone graft in the treatment of classⅡ furcations of mandibular molars.Methods: In the present study, thirty-five classⅡ furcation involvements were included and randomly divided into two groups. The experimental group (n=17) accepted GTR combined with CGF and bone graft therapy, and the controlled group (n=18) accepted CGF combined with bone graft therapy. The clinical examinations and cone beam computed tomography (CBCT) were performed at baseline and 1 year post-surgery. Comparisons of clinical and CBCT data before and after operation between the experimental group and the control group were made.Results: The clinical and CBCT data of both groups were not statistically different at baseline (P>0.05). At the end of 1 year post-surgery, the clinical parameters of both groups were significantly improved (P<0.001). The probing depths of the experimental group were (4.81±1.95) mm and (3.56±1.94) mm, respectively, significantly higher than the changes of the control group (P<0.001). The vertical and horizontal attachment gains of the experimental group were (4.11±1.98) mm and (3.84±1.68) mm, respectively, significantly higher than the changes of the control group (P<0.001). At the end of 1 year post-surgery, the experimental group showed significantly higher bone gain at vertical and horizontal directions compared with those of the control group: (3.84±1.68) and (3.88±2.12) mm, respectively (P<0.001).Conclusion: Within the limitation of the present study, GTR showed positive role in the effect of CGF combined with bone graft in the treatment of classⅡ furcation involvements of mandibular molars.

    Clinicopathological features and prognosis of colorectal stromal tumor
    Wen-peng WANG,Jie-fu WANG,Jun HU,Jun-feng WANG,Jia LIU,Da-lu KONG,Jian LI
    2020, (2):  353-361.  doi: 10.19723/j.issn.1671-167X.2020.02.025    
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    Objective: The incidence of colorectal stromal tumor is low among digestive tract tumors, therefore the literatures about clinicopathological features and prognosis of colorectal stromal tumor are few at home and abroad. In this study, we performed survival analyses for colorectal stromal tumor. The nomogram made by prognostic factors provided basis for evaluation of prognosis.Methods: The clinico-pathological and prognostic data of colorectal stromal tumor between January 1992 and December 2015 were collected from the surveillance, epidemiology, and end results (SEER) database. The survival analyses were made by SPSS 24.0 software. The nomogram and calibration curve were made by RMS package in R 3.5.2 software.Results: In the study, 546 patients with colorectal stromal tumor were included. The median age of onset was 64 years. The regional lymph node metastasis (LNM) rate was 9.4%. The multivariate Cox regression analyses of the 546 cases showed that the older age of onset (>64 years), single or divorce, colon tumor (compared with rectal tumor), non-surgery, high histological grade, LNM and distant metastasis were associated with worse cancer specific survival (CSS) and overall survival (OS), P < 0.05 for all. The treatment district was independent prognostic factor of OS (P = 0.027). The C-index of independent prognostic factors predicting CSS and OS probability were 0.76 (95%CI: 0.72-0.80) and 0.75 (95%CI: 0.72-0.78), respectively. Multivariate analyses were further carried out in the 174 patients with definite histological grade and tumor location, which revealed that the age of onset, histological grade, surgery or not were independent prognostic factors of CSS and OS (P < 0.05 for all). Tumor location was associated with CSS (P = 0.041) but not OS (P = 0.057) among the 174 cases. Four independent prognostic factors influencing the 174 patients' prognosis were used to make nomogram for predicting survival probability of 546 cases. The C-index of four prognostic factors predicting probability of CSS and OS of the 546 cases were separately 0.71 (95%CI: 0.66-0.75) and 0.73 (95%CI: 0.70-0.77). The nomogram had more accuracy for predicting OS probability of colorectal stromal tumors.Conclusion: The prognosis of colorectal stromal tumor was affected by multiple clinicopathological factors. The nomogram provided the basis for predicting the survival probability of patients with colorectal stromal tumor.

    Effect of convection enhanced delivery on the microstructure of brain extracellular space in aged rats
    Yu SONG,Hong-bin HAN,Jun YANG,Ai-bo WANG,Qing-yuan HE,Yuan-yuan LI,Guo-mei ZHAO,Ya-juan GAO,Rui WANG,Yi-xing HAN,Ai-lian LIU,Qing-wei SONG
    2020, (2):  362-367.  doi: 10.19723/j.issn.1671-167X.2020.02.026    
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    Objective: To compare the changes of extracellular space (ECS) structure and local drug distribution in adult brain and aged brain at different drug delivery rates in minimally invasive treatment of encephalopathy by convection enhanced delivery (CED) via ECS pathway.Methods: Thirty-six SD male rats were divided into adult rats group (2-8 months, n=18) and aged rats group (18-24 months, n=18) according to the age of the month. According to the drug rates (0.1 μL/min, 0.2 μL/min, and 0.3 μL/min), they were randomly divided into 3 subgroups, 6 in each subgroup. Gadolinium-diethylene triamine pentaacetic acid (Gd-DTPA) with a concentration of 10 mmol/L were introduced into the caudate nucleus of each group of rats by stereotactic injection. Tracer-based magnetic resonance imaging (MRI) was used to dynamically monitor the diffusion and distribution images of the Gd-DTPA in the brain interstitial system (ISS). Using the self-developed MRI image measurement and analysis system software to process and analyze the obtained images, the diffusion coefficient, clearance rate, volume fraction, and half-life of each group of rats in the caudate nucleus ECS could be acquired. The effects and differences of drug clearance and ECS structural function in the brain of aged rats and adult rats were compared and analyzed at different drug delivery rates. Magnetic tracer DECS-mapping technique was used to observe the distribution and drainage of tracer in caudate nucleus.Results: At the injection rate of 0.1 μL/min, the volume fraction in the aged rats was increased compared with that in the adult rats (18.20%±0.04% vs. 17.20%±0.03%, t=3.752, P=0.004), and the degree of tortuosity was decreased (1.63±0.04 vs. 1.78±0.09, t=-3.680, P=0.004), the drug clearance rate was decreased [(1.94±0.68) mm 2/s vs. (3.25±0.43) mm 2/s, t=-3.971, P=0.003], and the molecular diffusion in ECS was increased [(3.99±0.21)×10 -4 mm 2/s vs. (3.36±0.37)×10 -4 mm 2/s, t=3.663, P=0.004]. When the rate of injection increased to 0.2 μL/min, the drug clearance in ECS of the aged rats was slowed down [(2.53±0.45) mmol/L vs. (3.37±0.72) mmol/L, t=-1.828, P=0.021]. However, there were no significant differences in volume fraction,molecular diffusion in ECS and macroscopic drug metabolism parameters. When the rate of injection increased to 0.3 μL/min, the volume fraction in the aged rats was decreased (17.20%±0.03% vs. 18.20%±0.05%, t=-0.869, P=0.045), and the drug clearance rate in ECS was significantly accelerated [(4.04±0.76) mmol/L vs. (3.26±0.55) mmol/L, t=1.786, P=0.014], and there was no significant difference in tortuosity and the rate of molecular diffusion in the ECS. Conclusion: The drug clearance and ECS structural parameters of brain ECS in aged brain with CED administration were changed at different rates, and it has the least effect on ECS in the aged brain at the injection rate of 0.2 μL/min. For the application of CED for the treatment of encephalopathy, we should consider the influence of factors such as age and injection rate, and provide reference for the development of individualized clinical treatment plan for minimally invasive treatment of encephalopathy via ECS pathway.

    A novel tray for recording maxillomandibular relationship of edentulous patients based on dimensional surveying of complete dentures
    Yang LIU,Jin-ru HU,Yi-jiao ZHAO,Yong WANG,Yu-chun SUN,Shao-xia PAN,Hai-lan FENG
    2020, (2):  368-372.  doi: 10.19723/j.issn.1671-167X.2020.02.027    
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    Objective: To measure the dimensional data of complete dentures and to design a novel tray for recording maxillomandibular relationship of edentulous patients.Methods: For the measurement, 100 pairs of complete dentures from the clinic were surveyed for the following parameters: a1, the distance between the middle fossa of the upper left and right first molars; a2, the anterior-posterior distance between the middle fossa of the upper first molars and the incisal edge; a3, the width of the upper denture; a4, the anterior-posterior length of the upper denture; a51, the height from the mesio-lingual cusp of the right upper first molar to the saddle surface; a52, the height from the central fossa of the right lower first molar to the saddle surface; a6, the height from the notch of the upper lip frenulum to the upper central incisor edge; a7, the least thickness of the labial saddle base in the upper central incisor region. Based on the data, the trays with different sizes were designed and fabricated, and the key parameters were: b1, the distance between the foramina of screw posts, b2, the anterior-posterior distance between the foramina of the screw posts and the incisal edge; b3, the width of the tray; b4, the anterior-posterior length of the tray; b51, the height of the posterior platform with the screw nut; b52, the height of the screw post; b6, the height of the anterior tray handle; b7, the thickness of the anterior tray handle.Results: The minimum, average and maximum data for each parameter were (in millimeter): a1: 37.1, 44.5, and 59.6; a2: 22.6, 29.0, and 38.1; a3: 48.5, 58.2, and 76.6; a4: 37.4, 50.8, and 61.0; a51: 5.6, 9.5, and 14.7; a52: 3.8, 9.9, and 18.8; a6: 8.9, 16.6, and 24.7; a7: 1.2, 2.8, and 5.9. Based on the data, the trays in small, medium and large sizes were designed and fabricated. In clinical application, the putty silicone rubber impression material was used to reline the tray, meanwhile the posterior platform and anterior tray handle were set as the occlusal plane, then the screw posts were added and adjusted till the proper vertical dimension, after that, the putty silicone rubber impression material was added around the screw posts to record the horizontal maxillomandibular relationship, finally, the anterior surface of the tray handle was used to record the midline of the face and lower edge of the upper lip at rest and with smile.Conclusion: The dimensional data offered reference for the analysis of restoration space in edentulous patients. The tray designed and fabricated in this study may serve as a new tool for recording the maxillomandibular relationship.

    Kaposi sarcoma combined with severe ulcerative colitis: A case report and literature review
    Jun LI,Zhan-yue NIU,Yan XUE,Xue-ying SHI,Bo ZHANG,Yuan WANG
    2020, (2):  373-377.  doi: 10.19723/j.issn.1671-167X.2020.02.028    
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    SUMMARY Kaposi’s sarcoma (KS) is an unusual vascular tumor associated with human herpesvirus-8 (HHV-8) infection, which is common in immunosuppressors. Although extremely rare, iatrogenic (drug-related) KS can occur in human immunodeficiency virus (HIV)-negative patients under immunosuppressive therapy. We report a 64-year-old male diagnosed with ulcerative colitis for 1 year. He was treated with methylprednisolone because of an acute severe disease flare. He presented with several popular violet lesions on the body 4 months after steroid therapy. Histological examination of skin biopsies showed Kaposi’s sarcoma associated with HHV-8. The skin lesions regressed after steroid withdrawal and chemotherapy. Two key words “Kaposi’s sarcoma” and “inflammatory bowel disease” were searched in Wanfang data and CNKI, but no relevant articles were found. Thirty-eight articles in English were retrieved on PubMed with the key words of (“ulcerative colitis” OR “Crohn’s disease” OR “inflammatory bowel disease”) AND (Kaposi sarcoma). Twenty-five cases of Kaposi’s sarcoma related to inflammatory bowel disease (IBD) were reported. Including this case, the majority of 26 Kaposi’s sarcoma related IBD patients were male (80.8%, 21/26). The average age was (51.1 ± 16.4) years. Twenty cases were ulcerative colitis and 6 were Crohn’s disease. All the patients received immunomodulatory therapy, including glucocorticoid, azathioprine/mercaptopurine, methotrexate, cyclosporin and anti tumor necrosis factor α antibody. Thirteen cases were positive for HHV-8. There were 18 cases involving the distal ileum and colorectum only, 3 cases involving skin only, and 5 cases involving both skin and colorectum at the same time. Overall, the prognosis was good. Three patients only stopped immunosuppressive therapy, 1 received radiotherapy, 1 received chemotherapy, and 20 received surgery. Kaposi’s sarcoma could be seen in IBD patients with immunomodulatory therapy. It is very important to distinguish from the skin lesions related to IBD or drug treatment. The adverse reactions of immunomodulatory therapy should not be ignored. In addition, attention should be paid to the cooperation of multi-disciplinary team, which can diagnose and treat rare cases earlier and more accurately.

    Lower limb ischemia cured by stimulation electrode implantation assisted with microendoscopic discectomy system: A case report
    Ya-long QIAN,Shuai XU,Hai-ying LIU
    2020, (2):  378-381.  doi: 10.19723/j.issn.1671-167X.2020.02.029    
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    SUMMARY A 58-year-old male patient diagnosed with thromboangiitis obliterans (Fontaine stage Ⅳ) was recently treated with microendoscope discectomy system-assisted spinal cord stimulation electrode implantation and cured by department of vascular surgery combined with department of spinal surgery at Peking University People's Hospital. The patient suffered from cold injury to the right foot 14 years ago, which was cold, painful, numb, and then the toe was ulcerated and gangrene. Only the right foot small toe was left. The right foot skin was swollen from the toe to the proximal segment 1 year ago, accompanied by resting pain. Both pain and autologous bone marrow stem cell transplantation were ineffective. The above symptoms were aggravated three months ago, and the pain was severe. The visual analogue score was 10 points. A high amputation of the left lower extremity was performed 30 years ago due to trauma. Physical examination: the bilateral femoral artery was weak, and the right radial artery, posterior tibial artery, and dorsal artery were not touched. Buerger sign (+). Auxiliary examination: angiography of both lower extremities showed complete occlusion of the bilateral external iliac artery and its distal end. The percutaneous oxygen partial pressure was measured to be 30 mmHg on the right side of the iliac crest. The operation was performed under the local anesthesia. After X-ray positioning, the body projection of the lumbar vertebrae 1-2 lamina gap was marked. The skin had a 1.8 cm incision on the caudal side 2 cm from the mark. Then the dilators were used, and the working sleeve was tilted to the lumbar vertebrae 1-2 lamina gap. The microendoscope discectomy system was installed, the electrode was directly placed into the epidural space from the interlamina space under the microendoscope, the vascular surgeon adjusted the position of the electrode in the spinal canal under fluoroscopy, then connected the stimulator, adjusted the current until the patient had the lower limb fever, fixed electrode position, removed the microendoscope discectomy system after hemostasis under the microendoscope, used the guide needle to lead the electrode through the lumbar subcutaneous and then sutured the incision. After the operation, the electrode was connected to the temporary stimulator to stimulate for several minutes, the patient felt numbness in his lower limbs. In less than one hour, the skin temperature of the affected limb increased, and the painkiller could be stopped while sleeping. After 1 week, the skin temperature of the affected limb increased, and the percutaneous oxygen partial pressure of the foot and ankle was 36 mmHg, and the pain improved, and the score was reduced to 2 points. One month after surgery, the patient underwent permanent stimulator implantation. The pain disappeared after 3 months and half year of follow-up, and the score was reduced to 1 point. Microendoscope discectomy system-assisted spinal cord stimulation electrode implantation can complete the operation quickly, safely and effectively, and greatly reduce the number of intraoperative fluoroscopy and reduce the occurrence of complications.

    A rare case of cystadenoma in the small intestine
    Xiao WANG,Zhao-xing LI,Huan-fang FAN,Li-ying WEI,Xu-jin GUO,Na GUO,Tong WANG
    2020, (2):  382-384.  doi: 10.19723/j.issn.1671-167X.2020.02.030    
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    SUMMARY In recent years, there have been more and more reports about cystadenoma. Cystadenoma can occur in many parts of the body, and cystadenoma in different parts may show different clinical symptoms, however, some patients with cystadenoma have no symptoms. The vast majority of cystadenomas are benign lesions, but a small number of cystadenomas can be malignant. For example, a small number of ovarian cystadenomas and pancreatic cystadenomas may be malignant. This study reported a patient with small intestinal cystadenoma diagnosed by pathology. The patient's physical examination revealed a lesion in the left upper abdomen. He had only abdominal distension and no other discomfort. His laboratory examination results were basically normal, i.e. blood routine, urine routine, stool routine, liver function, kidney function, myocardial enzyme, tumor marker, etc. The patient underwent sectional small intestine resection and the pathological sample was analyzed. The histological findings of the resected intestinal sample were consistent with cystadenoma. Computed tomography scan of the abdomen was performed 4 months after the surgery. No recurrence of the tumor was found. The patient recovered in good condition. By consulting the literature, I found very few reports of small intestinal cystadenoma before, it was very rare. This article described the clinical manifestation, diagnosis and differential diagnosis, treatment and prognosis of a case of small intestinal cystadenoma, it suggested that cystadenoma can occur in the small intestine, other than the ovary, pancreas, liver, lung, thyroid, prostate, seminal vesicle, skin, etc. The cystadenoma in small intestine is easy to be mistaken with other tumors, such as small intestine stromal tumor, small intestine adenocarcinoma, small intestine lipoma, small intestine hemangiomas, etc., and it is difficult to fully confirm through imaging examinations, such as computed tomography and magnetic resonance imaging. Laparotomy and histopathological examination are necessary before definitive diagnosis. This disease can be treated by small bowel resection at the affected region and good prognosis can be achieved.

    Independent innovation research of functionally suitable denture digital system
    Yu-chun SUN,Yong WANG,Ke-hui DENG,Hu CHEN,Wei-wei LI,Yi-jiao ZHAO,Shao-xia PAN,Hong-qiang YE,Yong-sheng ZHOU
    2020, (2):  390-394.  doi: 10.19723/j.issn.1671-167X.2020.02.032    
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    SUMMARY Fabrication of conventional complete dentures involves a complex restoration method, requiring significant time and typically involving primary impressions, definitive impressions, jaw relation records, clinic try-in, and complete denture placement, which has been used for nearly a century without change. A novel digital system named Functionally Suitable Denture (FSD) was researched and deve-loped so as to reduce clinical steps, operation difficulties and errors of complete denture restoration. It pioneered a unique diagnostic complete denture aided by computer aided design (CAD) & 3D printing, by which, the functional impression, jaw relation, and try-in (3 steps) were simplified to 1 step, thus the number of visits to the dentist was reduced by 2 times. Moreover, for the first time, it put forward a CAD software of template matching based on the expert design, which was an efficient and intelligent design scheme, and the excellent denture experts' experience and skills could be inherited and iterated. The system included the 3D scanner with appropriate accuracy and high efficiency, the CAD software, the special 3D printer and process software, and the innovative clinical operation process. The Patent Cooperation Treaty (PCT) patent international search report showed that all the 15 claims of the technology were of novelty, creativity and industrial utility. All the digital products were independently developed and made by Peking University School and Hospital of Stomatology, China. The design and manufacture process of denture prosthesis was fast, simple and accurate. At the same time, personalized functional and aesthetic matching of the patients after wearing prosthesis was realized. It effectively solved the global problems of "slow, difficult and inaccurate" of the traditional manual technology of complete denture, and brought good news to edentulous patients. Compared with the traditional complete denture treatment, FSD system has a wide range of applications for different types of edentulous patients, including those with severe resorption of the alveolar ridge or a high occlusal force. Furthermore, the low-cost of 3D printers, compared with expensive milling machines, may make the approach more accessible. This review describes that our research is related to the development of the FSD system, including multi-source data acquisition technology, three generations of complete denture design software, 3D printing systems of individual tray and complete denture pattern, the clinical and laboratory operation process of the FSD system.


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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