Objective：To construct a new human chemotactic cytokine PSMP eukaryotic expression vector to express PSMP in Chinese hamster ovary (CHO) cells and to obtain the purified recombinant PSMP protein for its functional mechanism study. Methods: PSMP-myc/His fragment, cut from pcDNA3.1-PSMP-myc/His, was inserted into pMH3 expression vector. This expression vector was transfected into CHO cells by electroporation. Stable clone strains were selected by Geneticin resistance screening. The expressions of PSMP protein in the cell culture supernatant were measured by Dot blot and Western blot analysis. The monoclone was prepared from resistance screening polyclone by limiting dilution method. A large number of the engineering cells were cultured with serum-free medium and the protein in the cell culture supernatant was purified by nickel affinity chromatography. The purity of the PSMP protein was analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The functional activity of the protein was analyzed in vitro by Boyden chamber chemotaxis assay. Results:The eukaryotic expression vector pMH3-PSMP was successfully constructed by inserting PSMP-myc/His gene into pMH3 vector. After transfection of CHO cells, a stable expression of the PSMP gene engineering cell strain was obtained through twice cloning. The purity of the recombinant PSMP protein was 95% higher with bioactivity. Conclusion: The eukaryotic expression vector of PSMP protein is successfully constructed. The stable expression of PSMP is first obtained in CHO cell strain. The recombinant PSMP protein has higher purity and bioactivity, which provides a useful tool for further study of the functions and mechanisms of PSMP.

Objective：To compare the roles of single nucleotide polymorphism array (SNP array) and karyotype analysis in high-risk pregnant women prenatal diagnosis. Methods: From July 2012 to December 2013, a total of 141 pregnant women with high-risk in prenatal diagnosis were selected as the object of study in Department of Obstetrics and Gynecology, Peking University Third Hospital, 78 cases of umbilical cord puncture and 63 of amnion cavity puncture , both taking SNP array detection and karyotype analysis. Results: The abnormality karyotype rate was 6.4%, the abnormal rate of SNP array result was 11.3%, and the abnormal rate of the combined two methods for detecting was 12.1%. There were significant differences between the SNP array and karyotype analysis (P=0.039). Conclusion: There were obvious differences between the two techniques. It is an effective way to determine genetic disease by integrating SNP array and karyotype analysis in prenatal diagnosis.

Objective：To report the first case of fructose-1,6-bisphosphatase (FBPase) deficiency diagnosed by genetic sequencing in China, and to improve the cognition of this rare disease. Methods:The clinical and laboratory characteristics of FBPase deficiency were reviewed, and the findings of direct sequencing of genomic DNA described, and published literature on FBPase deficiency reviewed. Results: A 23-month-old boy was repeatedly admitted for 5 times with recurrent onset of lethargy and drowsiness every time after diarrhea and vomiting for 2－3 days during the last 7 months after being weaned, and he had convulsion this time. On admission, his physical examination showed tachypnea, and mild hepatomegaly, and he had normal physical and mental development. His paternal-grandparents had cousinship, and his parents were collateral relatives in the fifth generation. The laboratory findings revealed severe hypoglycemia, lacticacidemia, metabolic acidosis, ketonemia and hyperuricacidemia. After intravenous infusion of glucose, bicarbonate and antibiotics, there was a dramatic clinical improvement in a short time. Urine organic acids analyses ever showed an elevation of gluconeogenetic substrates including lactic acid, ketone and glycerol. The molecular analysis of liver fructose-1, 6-bisphosphatase (FBP1) gene showed a homozygous mutation with one G residue insertion at base 961 in exon 7(c.960/961insG), resulting in a reading frame shift mutation of 320th amino acid and premature termination at 333th amino acid. This mutation had been reported to be the most common mutation among patients with FBPase deficiency. Frequent feeding by avoiding taking in too much sweet food, restriction of food with high protein and fat, and the use of uncooked starch had been taken after our patient was discharged from the hospital. There had been no attack in the last 9 months. Conclusion: Clinicians must consider the diagnosis of FBPase deficiency when confronted with the patient who has episodes of severe hypoglycemia and lacticacidemia, especially accompanied by metabolic acidosis and ketonemia, which are typically triggered by infection and fasting. Early diagnosis, urgent treatment of hypoglycemia and appropriate diet control can prevent death, improve growth and quality of life of these children.

Objective： To study the evaluation of human epithelial growth factor receptor 2 (HER2) status in breast carcinoma with amplified chromosome 17 centromere locus (CEP17) and clinical significance of CEP17 amplification. Methods: Two hundred-eighteen cases of breast carcinoma were collected. We performed immunohistochemistry (IHC) to test HER2 protein and fluorescence in situ hybridization (FISH) to evaluate HER2 gene status. Results: Two cases in this cohort manifested CEP17 amplification. HER2 signals for case 1 was countable, and the average number was 2.6 per one nuclei, and the signals of CEP17 were clustered or multipunctiform. This case was evaluated as no HER2 amplification, but with amplified CEP17 . In case 2 the signals of HER2 and CEP17 were countable, and the average number of HER2 signal was 6.8 per one nucleus while CEP17 signal was 5.9 per one nucleus. The status was considered as HER2 and CEP17 coamplification. And the levels of HER2 protein expression of these two cases were both two plus. Conclusion： The incidence of CEP17 amplification in breast carcinoma is rare, with or without HER2 amplification. We recommend to evaluate the exact HER2 status by the HER2 copy number, and should also analyze the HER2/CEP17 ratio and the level of HER2 protein, for providing more accurate evidence to support the clinical target therapy.

Objective：To detect hot spot mutation of RYR1 gene in 15 cases of congenital myopathy with different subtypes, and to discuss the value of RYR1 gene hot spot mutation detection in the diagnosis of the disease.Methods: Clinical data were collected in all the patients, including clinical manifestations and signs, serum creatine kinase, electromyography. Fourteen of the patients accepted the muscle biopsy. Hot spot mutation in the C-terminal of RYR1 gene (extron 96-106) had been detected in all the 15 patients. Results: All the patients presented with motor development delay, and they could walk at the age of 1 to 3.5 years,but were always easy to fall and could not run or jump. There were no progressive deteriorations. Physical examination showed different degrees of muscle weakness and hypotonia.High arched palates were noted in 3 patients. The serum levels of creatine kinase were mildly elevated in 3 cases, and normal in 12 cases. Electromyography showed “myogenic” features in 11 patients, being normal in the other 4 patients. Muscle biopsy pathologic diagnosis was the central core disease in 3 patients, the central nuclei in 2 patients, the congenital fiber type disproportion in 2 patients, the nameline myopathy in 3 patient, the multiminicore disease in 1 patient, and nonspecific minimal changes in the other 3 patients; one patient was diagnosed with central core disease according to positive family history and gene mutation. In the family case (Patient 2) of central core disease, the c.14678G>A (p.Arg4893Gln) mutation in 102 extron of RYR1 was identified in three members of the family, which had been reported to be a pathogenic mutation. The c.14596A>G(p.Lys4866Gln) mutation in 101 extron was found in one patient with central core disease(Patient 1), and the c.14719G>A(p.Gly4907Ser) mutation in 102 extron was found in another case of the central core disease(Patient 3).The same novel mutation was verified in one of the patients’ (Patient 3) asymptomatic father.Conclusion: Congenital myopathies in the different subtype have the similar clinical manifestations, signs, enzyme detection and electromyography changes. Muscle biopsy plays an important role in the selection of genes to be detected. Hot spot mutation in C-terminal of the RYR1 gene can only be identified in patients with central core disease, so we suggest this hot spot gene mutation screening apply to the suspicious patient with central core disease only.

Objective：To screen and characterize the aptamer of Escherichia coli outer member protein tolC. Methods： By using the recombinant E.coli outer member protein tolC for the screening target, oligonucleotides which were capable of specifically binding to the protein were screened from a random oligonucleotide library through the stematic evolution of ligand by exponential enrichment (SELEX) technique. The binding capacity of ssDNA to the targeted protein from each round was detected by the FITC fluorescence labeling technique．The ssDNA from the last cycle was cloned and sequenced，and the second structure was further analyzed by the DNAMan program.  Results： After 12 cycles of selection, 40 clones were selected randomly and sequenced. Although a unique conserved sequence was not obtained among the 23 obtained aptamers by the primary structure analysis，three pairs of aptamers and two pairs of aptamers were found to be identical．Analysis of the secondary structure revealed that the stemloop and bulge loop were the main motifs，indicating that they might play a key role in the binding of aptamers to the target protein. According to the characteristic of the second structure，23 aptamers were divided into four families，and aptamer 20 bore the greatest affinity.Conclusion： Aptamers against E.coli outer member protein tolC were successfully identified by the SELEX method. The results laid a foundation for the investigation of the interference to the drug resistance of E. coli and the underlying mechanisms.

Objective：To observe the multiple influence of cholesterol-lowering drug (simvastatin) on ankle brachial index (ABI), flow-mediated dilation (FMD) and nitroglycerin-mediated dilation (NMD) of brachial artery blood vessel endothelium, and plasma level of monocyte chemotactic protein 1 (MCP-1) of hypercholesterolemia patients without coronary heart disease (CHD). Methods：In the study, 51 patients with hypercholesterolemia application were treated with simvastatin (20 mg/d) therapy for 12 weeks. The metabolic index, ankle brachial index (ABI), FMD of brachial artery blood vessel endothelium detected by color doppler ultrasound instrument, the NMD of artery endothelial and the level of MCP 1 were measured before and after therapy respectively. All the results were analyzed and compared with another 30 cases of hypercholesterolemia patients selected without simvastatin treatment. Results：After simvastatin therapy, the TC (total cholesterol) and LDL-C (low density lipoprotein cholesterin) levels were reduced apparently，the values decreased from the original (6.06±1.03) mmol/L and (3.60± 0.82) mmol/L to (4.98±1.34) mmol/L and (3.41±0.10) mmol/L respectively (P<0.01, P< 0.05). Compared with no simvastatin treatment, the bilateral ABI levels were significantly elevated. The right side of ABI (ABIR) elevated from 1.11±0.06 to 1.19±0.07, and the left side of ABI (ABIL) also elevated from 1.12±0.06 to 1.19±0.10 (both sides were P<0.01). The FMD significantly increased from 7.75%±11.30% to 14.20%±15.39% (P < 0.05). The plasma levels of MCP-1 were apparently reduced from (112.0±7.8) ng/L to (108.9±6.2) ng/L (P < 0.05). All these items showed no obvious change within the control group. Conclusion：The API, FMD and plasma levels of MCP-1 of hypercholesterolemia patients without clear coronary heart disease can be improved by simvastatin treatment.

Objective： To investigate the diagnostic value of serum tumor marker in solitary pulmonary nodules (SPN). Methods: In the study, 175 cases of SPN postoperatively diagnosed by pathology between Jan. 2011 and Jan. 2013 in Peking University First Hospital were selected, including 125 cases of lung cancer and 50 cases of benign lesions. The levels of serum carcino-embryonic antigen (CEA), squamous cell carcinoma (SCC), neuron-specific enolase (NSE) and cytokerantin-19-fragment (CYFRA21-1) were detected by electrochemical luminescence immunoassay. SPSS 11.5 software package was used for statistical analysis. Results: In the malignant SPN group, CEA, SCC and CYFRA21-1 levels were significantly higher than in the benign group (P<0.05). The positive rates of CEA, SCC, CYFRA21-1 and combined detection of the four serum tumor markers in the malignant SPN group was significantly higher than in the benign group (P<0.05). ROC curves showed that the under-curve area of CEA, NSE, SCC and CYFRA21-1 was 0.633±0.045, 0.494±0.047, 0.664±0.042 and 0.711±0.041, respectively. The combination of CEA, SCC, NSE and CYFRA21-1 showed the highest sensitivity (52.0%) and better specificity (76.0%) for diagnosis of lung cancer. There were no statistical differences in the positive rates of tumor markers between the malignant SPN group and benign group in the smoking patients (P>0.05). Conclusion: CEA, SCC and CYFRA21-1 have higher positive rates in the malignant SPN patients, suggesting a certain value in the early diagnosis of malignant SPN.

Objective：To improve the diagnosticability of selective enteroclysis in the area of intestinal obstruction, by reviewing the cases of selective intestinal enteroclysis through a naso-intestinal decompression tube, and evaluating its diagnostic concordance level and the influence factors. Methods: The selective enteroclysis was different from traditional enteroclysis; it utilized a naso-intestinal decompression long tube to perform local intestinal opacification. In the four years between Apr. 2008 and Apr. 2012, 98 cases of selective intestinal enteroclysis were analyzed retrospectively, and diagnostic concordance value was made between the radiologic diagnosis and the final clinical diagnosis recorded in the medical history according to an evaluating criterion. Five scores were used in the evaluating system: 0 meant that the enteroclysis had no valuable information to provide; 0.25 meant that the enteroclysis could prompt the occurring of obstruction, but could not provide the information of location and cause; 0.50 meant that the enteroclysis could find the location of obstruction, but could not determine the cause; 0.75 meant that the enteroclysis could provide some valuable analysis of the causes of the obstruction, and very close to the final clinical diagnosis; 1.00 meant that the complete concordance between the enteroclysis diagnosis and the final clinical diagnosis. The influence factors would also be considered to improve the competence of the selective enteroclysis in the diagnosis of intestinal obstruction. Results: In the 98 cases, 53.1% had definite abdominal operative history. The most common cause of obstruction was conglutination between intestinal loops when discharged from hospital, almost occupying 50%. In this group of cases, the most common types were ileum obstruction (47.6%), multiple location obstruction (41.7%) and incomplete obstruction (59.5%). Conglutination and stricture of the intestinal was the common radiologic appearance (61.9%). There were 50 cases with higher concordance scoring 0.75 or 1.00, in comparison, there were 25 cases with lower concordance scoring 0 or 0.25. The difference between the two groups had statistical significance. Conclusion: As a combination of traditional enteroclysis and nasointestinal decompression tube, selective intestinal enteroclysis could exert higher diagnostic ability than that of traditional enteroclysis and also could break through the limitations of traditional enteroclysis in the condition of intestinal obstruction. This method has higher diagnostic concordance and could provide valuable information in obstruction location, extent, severity and possible causes. The main factor influencing the effectiveness of the examination is the location of the decompression tube and dynamic observation would be very helpful and important.

Objective：To establish the characteristics of uncomplicated acute appendicitis (UAA) and assess the efficacy of antibiotics by comparison with emergency appendicectomy for treatment. Methods: A retrospective clinical analysis was made on 742 cases of acute appendicitis. Several characteristics of UAA were identified. Following these rules 46 UAA patients were enrolled to receive antibiotic treatment for at least 48 h.Follow-up was carried out for 6 months.Results: Statistical analysis indicated that high level white blood cell count (>20×10⁹/L,P=0.000, OR=2.717), local or diffuse muscle guarding (P=0.031, OR=1.649), intraluminal stercolith (P=0.000, OR=2.939) and periappendiceal fluid (P=0.005, OR=3.273) were independent risk factors of complicated acute appendicitis. With none of these factors we enrolled 46 UAA patients. Of the 46 patients, 44(95.65%) were treated successfully with antibiotics. 2(4.35%) patients were unexpectedly identified to have complicated appendicitis at surgery. Recurrent appendicitis occurred in 5(11.36%) patients after 6 months. There was no difference in duration of pain and duration of hospital stay between antibiotic treatment group and appendicectomy group. Conclusion: Although antibiotic treatment may fail in some UAA cases, and there is a risk of recurrence, antibiotic treatment still appears to be a safe firstline therapy for UAA patients.

Objective： To evaluate the treatment outcome of the “All-on-4” immediate loading protocol via survival rate of the implants，survival rate of the prosthesis，marginal bone, postoperative complications and patient satisfaction.Methods: In our study, 40 patients with 49 edentulous jaws (31 mandibles and 18 maxillae) were enrolled. Each jaw was restored by the shortened dental arch prosthesis supported by only 4 implants according to the All-on-4 protocol (All-on-4, Nobel Biocare AB, G-teborg, Sweden). For all the patients enrolled in the study,the loading was applied within 12 hours of surgery. The provisional prosthesis could be replaced by the final restorations within 6 to 12 months. In the present study, the survival rate of the both implants and restorations were calculated and analyzed. The radiographic evaluation of marginal bone level changes was measured. The values of the marginal bone level changes of the angled and axial implants were analyzed by the statistic software.Results: In the present study, totally 196 implants were inserted, of which 13 implants failed during the whole following up periods, with 11 implants of the maxillae and 2 of the mandibles. The survival rate of the prosthesis was 95.9% (47/49). The implant survival rate of the maxillae was 85.5%（65/76）while that for the mandibles was 98.3%(118/120). The implant survival rate of the angled implants was 91.8% (90/98), while that for the straight implants was 95.0% (93/98). No significant difference in marginal bone loss was found between angled and axial implants in the 12-month evaluation according to the Wilcoxon rank sum test (P>0.05). During the follow-up period，mechanical complications as fracture of the provisional prostheses, loose of the retain screw, or crack of the artificial teeth were found in 20 prostheses.Conclusion: The present preliminary data of the short term observation suggest that the “All-on-4” immediate loading protocol is a viable treatment modality for the edentulous jaws. However, long term clinical random controlled trials with large samples are still needed to confirm the validity of the technique.

Objective：To evaluate the effect of regenerative therapy for the treatment of furcation involvements of mandibular molars with cone-beam computed tomography (CBCT). Methods: In the study, 38 furcation involvements of mandibular molars were included and randomly divided into two groups. The experimental group accepted guided tissue regeneration and bone graft therapy, and the control group only flap surgery. The clinical examination and CBCT examination were performed at baseline and 1 year post-surgery. 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, except gingival recession, the clinical parameters of both groups were significantly improved (P<0.001). The vertical and horizontal attachment gains of the experimental group were (3.20±1.82) mm and (2.05±1.27) mm, respectively, and significantly higher than the changes of the control group (P<0.001). And at the end of 1 year post-surgery, the experimental group showed significantly higher bone gain at the vertical and horizontal directions compared with those of the control group: (2.82±0.97) mm and (2.24±0.92) mm, respectively (P<0.001). Conclusion: With the limitation of this study, the effect of guided tissue regeneration surgery and bone graft therapy for the treatment of furcation involvements of mandibular molars is significantly better than that of the flap surgery. CBCT can reflect the horizontal and vertical bone changes of furcation area, which is more comprehensive than traditional periapicals.

Objective：To develop and study the properties of crosslinked polyvinyl alcohol microspheres (PVA-Ms) for embolization. Methods: The PVA-Ms were produced by emulsion chemical crosslinking method. Fourier transform infrared spectroscopy (FT-IR) was used to investigate the special functional groups of PVA-Ms; the morphology and particle size of PVA-Ms were determined by optical microscope; the ratio of water absorption and the swelling ratio were also investigated; the compressibility was examined by texture analyzer. A new device was designed to measure the pressure of PVA-Ms during their delivery through catheter for embolization. Results: The crosslinking reaction of PVA and formaldehyde was proved by FT-IR. The PVA-Ms were round with smooth surface. The average diameter of lyophilized PVA-Ms was 574.2 μm with a range of 80~1 800 μm and of wet PVA-Ms was 602.2 μm with a range of 100~1 900 μm. The average ratio of water absorption was 175% and the swelling ratio was 48.6%. The PVA-Ms were mechanically stable with appropriate elasticity and delivered through the catheter without any difficulty, and the pressure was higher for larger size of microspheres to be delivered. Conclusion: PVA-Ms prepared in this study was supposed to be suitable for clinical embolization according to the physicochemical properties. The study provides a series of methods to evaluate the properties of microspheres systemically for embolization in vitro.

Objective：To propose a calculation method of oligosaccharides’ fractal dimension, and to provide a new approach to studying the drug molecular design and activity. Methods: By using the principle of energy optimization and computer simulation technology, the steady structures of oligosaccharides were found, and an effective way of oligosaccharides fractal dimension’s calculation was further established by applying the theory of box dimension to the chemical compounds. Results: By using the proposed method, 22 oligosaccharides’ fractal dimensions were calculated, with the mean 1.518 8±0.107 2; in addition, the fractal dimensions of the two activity multivalent oligosaccharides which were confirmed by experiments, An-2 and Gu-4, were about 1.478 8 and 1.516 0 respectively, while C-type lectin-like receptor Dectin-1’s fractal dimension was about 1.541 2. The experimental and computational results were expected to help to find a class of glycoside drugs whose target receptor was Dectin-1. Conclusion: Fractal dimension, differing from other known macro parameters, is a useful tool to characterize the compound molecules’ microscopic structure and function, which may play an important role in the molecular design and biological activity study. In the process of oligosaccharides drug screening, the fractal dimension of receptor and designed oligosaccharides or glycoclusters can be calculated respectively. The oligosaccharides with fractal dimension close to that of target receptor should then take priority compared with others, to get the drug molecules with latent activity.

Objective：To develop and investigate the properties of MRI-traceable Eudragit-E liquid embolic agent (MR-E). Methods: Polyethylene glycol-modified superparamagnetic iron oxides (PEG-SPIO) was synthesized by chemical co-precipitation method. MR-E was prepared by mixing PEG-SPIO and Eudragit-E liquid embolic agent homogeneously. An in vitro MR phantom study was carried out to measure MR traceability of MR-E and to determine the concentration of PEG-SPIO for further studies. The microcatheter deliverability and sol-gel transition process of MR-E were investigated. MR-E was injected into the kidney of a Japanese white big ear rabbit via an angiographic microcatheter, and detected by MRI. Results: A PEG-SPIO concentration of 2 g/L was considered to be suitable for further studies. MR-E was injected through the microcatheter without any difficulty. MR-E instantly solidified on release into saline. Then 0.2 mL of MR-E effectively embolized distal renal arteries, and MR-E could be detected by MRI in the embolized kidney. Conclusion: MR-E seems to be a promising MRI-traceable liquid embolic agent.

Objective：To observe the characteristics of acoustic parameters of 21 patients with amyotrophic lateral sclerosis (ALS) complaining of dysarthria and to explore the possibility and clinical value of Multi-Dimensional Voice Program (MDVP) on aseessing the patient’s voice. Methods: The clinical data and speech intelligibility of each of the 21 patients with ALS complaining of dysarthria were collected. All the 21 patients with ALS and 44 normal subjects were suggested to pronounce the vowel ［a：］.The voice samples were collected and analyzed by Multi Dimensional Voice Program. The data were compared to detect the difference between the two groups. Results: Speech intelligibility of 10 in the 21 patients was above 97% including 5 patients whose scores were 100%. It seemed that the speech intelligibility score was not related with the duration of the disease and the onset part. The values of noise to harmonic ratio (NHR) of both the light and the severe damage to the speech group were higher than those of the control group with significant difference （P<0.05）.Significant difference was noticed between the severe damage to the speech group and control group in Jitter. Conclusion: Multi Dimensional Voice Program could be used to analyze the characteristics of acoustic parameters of patients with ALS patients complaining of dysarthria. There was more noise composition in the voice of the patients with dysarthria. NHR seemed to be the sensitive and stable parameter. Especially in patients whose speech intelligibility were affected severely, more acoustic parameters were detected with significant difference.

Objective：To explore the protected effect of sensory baby-sitter in reverse end-to-side fashion on denervated muscle. Methods: The tibial nerve of twelve female adult Sprague Dawley rats was transected. Six animals served as controls. In the other rats, the end of the sural nerve was connected to the side of the distal tibial nerve stump. After twelve weeks, the wet weight, cross-sectional area, motor endplate perimeter from gastrocnemius muscle were examined. Results: The difference in wet weight between the experimental group and the control group was statistically significant (39.2%±6.8% vs. 19.5%±4.3%, P<0.05). Histological observation of the unprotected muscles displayed wide areas of atrophied fibers and considerable connective tissue hyperplasia, whereas the structure of the experimental rats was preserved and there was only a slight increase in connective tissue. The average crosssectional area and motor endplate perimeter of muscle fibers were significantly larger in the experimental group than in the control group ［(1 148.85±547.18) μm2 vs. (575.05±140.51) μm2, (102.84±53.29) μm vs. (59.60±26.71) μm, respectively］. Conclusion: Sensory babysitter in reverse end-to-side neurorrhaphy preserves the structure of denervated muscle in rats.

Objective：To evaluate the effect of chondrocyte mitochondrial dysfunction on the development of cartilage degeneration. Methods: In the study, 10 cartilage samples of the knee joint were collected during total knee arthroplasty surgery because of OA from April to October of 2012 in Peking University First Hospital. All the tissues were taken from transmission electron microscope (TEM) observation grouped by Outerbridge classification. Then, TEM observation, quantitative detection of mitochondrial respiratory chain enzyme complex 1,2,2+3,4 and ATPase activity, detection of the mitochondrial membrane potential by JC-1 method were taken with cultured normal and OA chondrocytes. Healthy chondrocytes from 10 normal cartilage samples were divided into 2 groups: the normal control group and rotenone group. The ultrastuctrure alterations of mitochondria, mitochondrial membrane potential, apoptosis rate and collagen Ⅱ content were compared. Results: With the aggravation of cartilage degeneration, mitochondria swelling, outer membrane rupture, cristae destruction and disappearance were observed in both the tissue and cell TEM examinations. JC-1 staining showed a decreased membrane potential in OA chondrocytes which had a lower red/green fluorescence ratio of 1.50 than that of the normal chondrocytes of 2.58. mitochondrial respiratory chain (MRC) enzyme complex 1,2,2+3,4 and ATPase activity of the OA chondrocytes also represented a decreased tendency compared with the normal chondrocytes although the difference was not significant (P=0.109,0.197,0.098,0.169,0.145). The mitochondria in the Ro group cells showed OA-like changes morphologically by TEM detection. JC-1 staining showed a decreased mitochondrial membrane potential in the Ro group chondrocytes which had a lower red/green fluorescence ratio of 1.78 than that of the normal ones of 2.58. Apoptosis examination represented a higher apoptosis rate of 7.53% in the Ro group chondrocytes than that of the normal ones of 4.38%. Collagen Ⅱ content of the chondrocytes in the Ro group was (44.63±7.11) μg/L , significantly lower than (72.88±24.3) μg/L in the control group (P=0.044). Conclusion: Mitochondrial function is impaired in OA chondrocyte. Mitochondrial function destruction results in an increased chondrocyte apoptosis rate and a decreased collagen Ⅱ secretion.

Objective：To investigate the surgical treatment results of implant failure after clavicular fracture open reduction and internal fixation (ORIF). Methods: Fifteen cases from Jan. 2005 to Jan. 2013 were treated surgically according to fracture classification, time of implant failure and implant type. The fracture union, shoulder function and pain were evaluated postoperatively. Results: All the patients had full follow-up for 5 to 101 months (mean: 43.8 months). All the fractures were united well. The constant scores to assess the shoulder function were 82 to 100 (mean: 93.3 in the fracture side) and were 85 to 100 (mean: 96.7 in the uninjured side); statistically significant difference of the constant scores between the two sides was found (P=0.02). Eight cases did not have shoulder pain in the fracture side, while the other 7 cases had mild pain, The visual analogue scale (VAS) scores to evaluate shoulder pain were 1 to 3 in the fracture side, which were statistically different from those in the uninjured side (P=0.03). Conclusion: Implant instability causes early implant failure after clavicular fracture ORIF and refixation with stable implant is effective. Fracture nonunion leads to late implant failure, and bridging fixation using locking plate associated with bony autograft with iliac crest is a successful method to treat atrophy clavicular nonunion. Surgical treatment can bring good results.

Objective：To examine pathological changes in different parts of the larynx in canines after laryngeal burns induced by hot dry air at various temperatures and compare the pathological results with clinical typing. Methods: Eighteen healthy, male, adult dogs were randomly assigned to inhale hot dry air at 80 ℃ (Group 1), 160 ℃ (Group 2) or 320 ℃ (Group 3) for 20 min to induce inhalation injury. Pathological changes after hot air inhalation were examined at different locations of the larynx including the epiglottis, laryngeal vestibule, vocal folds and trachea. Results: Pathological changes were mainly characterized by local mucosal atrophy in the larynx in Group 1 and moderate edema in the laryngeal submucosa in Group 2. Group 3 showed two types of pathological changes in the larynx: severe edema as well as atrophy and charring. Conclusion: Due to its special anatomy and functions, the larynx has different responses to inhalation injuries induced by hot air at different temperatures. The pathological observation and analysis showed that the pathological changes brought about by laryngeal burns at 80 ℃ and 160 ℃ were generally consistent with laryngoscopic manifestations of congestion and edema. Inhalation of hot dry air at 320 ℃ led to two types of pathological changes, severe edema and atrophy and charring. Hence, pathological analysis of laryngeal burns may be used as a supplemental tool to clinical typing.

Objective：To evaluate the current condition of urban road traffic injuries (RTIs) according to Beijing Emergency Medical Center （BEMC） from Jan. 1, 2004 to Dec. 31, 2010, analyze the social characteristics and explore the possible methods for prevention and improvement. Methods：Using data from the Beijing Emergency Medical Center, we collected 19 550 victims who were involved in RTIs in Beijing from Jan. 1, 2004 to Dec. 31, 2010. The personal information, time of the injury event, road user type and striking vehicle type, as well as the site and severity of injury, were analyzed using Excel 2007 and SPSS 17.0 software with ANOVA of variance and Chi-squared tests. Results：The annual rate of RTIs was 120.0 per 100 000 people in Beijing, and the mortality rate was about 4.97 per 100 000 people. Male victims were more than female victims (11 737 persons vs. 7 618 persons).The mean age was (72.92±5.67) years. Overall, RTIs in all the age groups happened in October commonly, and were inclined to daytime, especially at noon. But different age groups had their special hour distribution features of RTIs. Traffic collisions occurred most frequently in pedestrians and cyclists（7 588,38.81%；3 790,19.39%）. Majorities of victims presented with head injuries and lower-limb injuries（8 343,42.68%; 6 828,34.93%）. These collisions included car striking accidents (11 490, 58.77%). And most of the older adults were classified as medium in severity (11 718, 59.94%). Conclusion：The prevention and treatment of RTIs, should focus on targeted prevention solutions and standardized pre-hospital rescue, according to specific population, time interval and vehicle usage.

Objective： To provide scientific evidence for medical insurance and health policies allocating the limited health resources in China. Methods: Based on the data of the national household survey by the State Council Pilot Urban Resident Basic Medical Insurance Evaluation from Nov.2007 to Nov.2011, a twostep model and the human capital method were used to estimate the economic burden of five major chronic diseases among urban residents in China. According to the economic burden, the relative economic risk (relative risk, RR) and adjusted RR were calculated. The five chronic diseases were hypertension, cardiovascular, diabetes, arthritis or rheumatism and chronic lung diseases. Results: More than 50% of the residents with these five chronic diseases were the over 65-year-old and retired. 90% of the residents with these five chronic diseases had medical insurance except the residents with chronic lung diseases. Average co-pay from the outpatient department and the pharmacy was more than 60%, and about 50% from the inpatient department. Annual total cost per capita was the highest 8 954.29 Yuan among the residents with cardiovascular disease and the second highest 8 914.36 Yuan among the residents with diabetes. The adjusted RR of the residents with cardiovascular and diabetes were greater than 1, respectively 1.36 and 1.15. Conclusion: The retired take up the largest percentage of population with chronic diseases, and the influence of the major five chronic diseases is more serious in north-west China. The main expenditure is from the outpatient department and the pharmacy, in which the availability of drugs reimbursed needs to be improved. The patients with cardiovascular and diabetes experience both higher economic burden and economic risk.

Objective：To study the characteristics of histopathologic features of small renal carcinoma. Methods: This retrospective study collected the data of renal cell carcinoma from the patients who underwent surgery from January 2002 to June 2012. They were all preoperatively diagnosed as renal cancer by CT scan, and pathologically diagnosed as renal cell carcinoma after surgery with the diameter ≤4 cm. We recorded and analyzed the tumor size, histologic subtype, Fuhrman grading, TNM stage, the existence of tumor vascular invasion, sarcomatoid differentiation, and whether it was multifocal, and then grouped them for comparison. Results: A total of 1 276 patients were included in the study and were analyzed, of whom 306 (24.0%) had small renal cell carcinoma less than 2.0 cm, 526 (41.2%) 2.0-3.0 cm, and 444 (34.8%) bigger than 3.0 cm. Of all the subjects, 1 158 (90.7%) suffered from clear cell carcinoma, 49 (3.8%) papillary carcinoma, 32 (2.5%) chromophobe cell carcinoma, whose distribution was not related with tumor size. Of the ≤2.0 cm and >2.0 cm groups, Furmann grades of G3/4 were 15 (4.9%), 98 (10.1%), respectively (P=0.007). T3a+ stage and above were 1 (0.3%), and 32 (3.3%), respectively (P=0.004). Synchronous distant metastases occurred in 6 patients, all in group >2.0 cm. Tumor sarcomatoid differentiation (0.3% vs. 0.9%), vascular invasion (0.6% vs. 2.1%) and multifocal (1.3% vs. 2.7%) had no significant difference between the two groups. Conclusion:Small renal cell carcinoma with diameter over 2.0 cm are more aggressive, suggesting that renal cancer bigger than 2.0 cm in diameter should not select a nonsurgical treatment.

Objective： To find the causes of retrograde intrarenal surgery (RIRS) failure in renal calculi treatment and evaluate the efficacy and morbidity of remedial measures.Methods: All the patients with renal stone who had undergone RIRS in the last two years were reviewed and the failure cases screened out according to operating records. Results: Within the 132 patients, 19 RIRS procedures failed. The causes of failure included ureteral (42.1%), intrarenal(36.8%), stone(10.5%) and operating reason(5.3%).The caculi of 11 patients were cleaned by remedial percutaneous nephrolithotomy (PCNL), seven patients had secondary RIRS one week later, and only one patient left the renal stone for observation and dietary regulation. Sixteen patients were stone-free after remedial treatment and the complications included ureteral perforation in one case and high fever in 4 cases.Conclusion: The main causes of RIRS failure are ureteral incompatibility and intrarenal disadvantageous anatomy. PCNL and secondary RIRS are the effective remedial measure after RIRS failure.

Objective： To investigate the bacteriology and drug sensitivity of upper urinary tract calculi patients, and to provide information for choosing suitable antibiotics. Methods： In the study, 21 patients who suffered from lithiasis in upper urinary tract and required an emergency drainage for acute obstruction and infection were the “acute group”; 64 patients with calculi in upper urinary tract and accompanied with no infectious symptoms were the “common group”. The bacteriology and drug sensitivity of the two groups were investigated. Results： Gram-negative bacteria infected the most common of upper urinary tract calculi patients with infection, accounting for 71.4% in the acute group and 65.7% in the common group, among which Escherichia coli were the predominant ones (35.7% in the acute group and 32.9% in the common group). No difference was found between these two groups in bacterial distribution (P>0.05). Although the average drug resistance rate of Gram-negative bacteria in the acute group was higher than that in the common group, it revealed no significant difference (P>0.05). The drug resistance rate to semisynthetic penicillin, cefuroxime and ceftriaxone were more than 50%, 60%, and 50%, respectively. Quinolones, such as ciprofloxacin and levofloxacin, got a 45% drug resistance. Aminoglycoside, carbapenema were sensitive to Gram-negative bacteria. Cefoperazone/sulbactam and piperacillin/tazobactam were more effective than ceftriaxone and piperacillin, respectively. Conclusion： There was no significant difference between upper urinary tract calculi patients with acute infection and common infection in bacteriology and drug sensitivity. Semisynthetic penicillin, the second generation of cephalosporin and quinolone were no longer the good choices of empirical use. Antibiotics combined with β-lactamase inhibitors would be an ideal empirical therapeutic choice.

Mutations in the fibrinogen Aα-chain genes are the most common cause of hereditary renal amyloidosis. The renal histologic appearance in the patient is characteristic and shows striking glomerular enlargement with almost complete obliteration of the normal glomerular architecture by extensive amyloid deposition. In contrast, the vessels and renal tubular interstitium of such patient contains almost no amyloid at all. Here, we described a patient with hereditary fibrinogen amyloidosis, who presented with proteinuria, hypertension and renal failure. He was shown to be heterozygous for the relevant mutation encoding the E526V fibrinogen variant.

Autoimmune diseases can cause various kinds of lung injuries. Clinical features of a case of overlap syndrome with multiple pulmonary injuries were investigated, and the treatment experiences discussed. The patient suffered from rheumatoid arthritis (RA) at first, and then had a lobectomy surgery due to the rheumatoid nodules in her right lung. A year later her disease was diagnosed as amyopathic dermatomyositis (ADM) with typical Gottron’s sign, craftsmen hands and rapidprogressive organizing pneumonia (OP). After a combined treatment with glucocorticoids (GCs) and cyclophosphamide (CTX), her OP became better but lung infections progressed. Her lung infections eventually were cured after we used antibiotics and antifungal treatment while we ceased to use CTX and reduced the dosage of GCs. The clinical feature of the patient was overlap syndrome with a variety of lung injuries, such as pulmonary rheumatoid nodules, OP, secondary bronchopleural fistula and lung infections. Its diagnosis and treatment experiences could improve our understanding of pulmonary manifestations of connective tissue disease and improve our diagnosis and treatment level.

Warfarin is a high-alert medication, which may result in bleeding if used improperly. In our case, one elderly female with atrial fibrillation had taken warfarin for more than half a year, and her international normalized ratio (INR) was maintained within the therapeutic range. The patient began to take tramadol to alleviate her shoulder pain. Three days later she presented hematuresis and had ecchymosis in her right upper arm, and in the meantime her INR rose to 10.04. Clinical pharmacists analyzed the cause for bleeding by searching relevant literature, and finally discovered the interaction between warfarin and tramadol. On the basis of that, the clinical pharmacists provided pharmaceutical care, offered specific medication education, as well as assisted the physicians to establish the medication plan for warfarin reuse. Eventually, her INR declined to reference ranges, and her hematuresis and ecchymosis were alleviated significantly. This successful case reveals that clinical pharmacy services contribute to better treatment outcomes. Clinical pharmacists can play an active role in anticoagulation management in healthcare team.