Objective To investigate the dynamic changes of interleukin-1 receptor-associated kinases (IRAK-4) in hypoxic neurons and explore their role in regulation of inflammatory reaction. Methods The B35 cells exposed to hypoxia of 3% O2,5% CO2 and 92% N2 were cultured for 1,3,6, 12,24,48,72 and 96 hours respectively. Then, mRNA and protein expressions of IRAK-4 were detected by RT-PCR and Western blot, the expression of IRAK-4 in the cells were observed by laser scanning con-focal microscope (LSCM), and the concentration of IL-6 was measured by ELISA method. Results After hypoxia, the mRNA and protein expressions of IRAK-4 were increased at one hour, reached the peak at six hours (P<0.05), kept at a high level at 12 hours (P<0.05) , but decreased gradually to the normal oxygen level at 24 hours (P < 0.05) and to below the normal oxygen level at 48 hours (P < 0.05). Immunofluorescence results showed that the fluorescence intensity of IRAK-4 was gradually increased with time. The changes of IL-6 in the supernatant were positively correlated with protein expression of IRAK-4 (r =0.84, P < 0.05 ). Conclusions Hypoxia can increase the expression of IRAK-4 at transcription and translation levels in a certain period of time, which may participate in down-stream inflammatory reaction and lead to increase of IL-6 expression.

Objective To investigate and analyze the reasons of fa0ilure in external quality assessment(EQA) for routine chemistry and provide the basis for the corrective and preventive actions.Methods Based on the network system of NCCL EQA the reasons of failure in 2013 national routine chemistry external quality assessment program were investigated,among which the reasons were classified and analyzed with seven sources of problems which were clerical errors,methodological problems,equipment problems,technical problems,EQA materials problems,EQA Evaluation problems and unable to explain after investigation.Results The return rate of this root cause investigation for each analyte ranged from 33.3％ to 80.0％.The major reason for unacceptable analyte included clerical errors (6.5％) (decimal point position error:70.1％;unit error:20.8％;instrument or method coding error:8.1％),methodological problems (45.1％)(calibration:54.2％;reagent:38.0％;EQA material:7.8％),equipment problems (28.5％) (no regular maintenance:98.0％;pipeline error:2.0％),technical problems (8.2％) (do not follow SOP:80.4％;EQA material redissolve error:10.6％;placing order error:9.0％) and unable to explain (11.7％) (system error:68.2％;random error:31.8％).There were no EQA materials problems or EQA Evaluation problems in this survey.Analysis systems' grouping statistics were implemented for seven analytes including sodium,chlorine,phosphorus,direct bilirubin,total iron binding capacity,copper,and zinc.Unsatisfied EQA proportions of mating system were lower than nonmatching ones for the majority of analytes.Conclutions Further work on EQA should be undertaken by clinical laboratories.Laboratories should use reagents with high quality as well as improve the operation technology and sense of responsibility.Only in this way,can the accuracy and reliability of testing results be guaranteed.

Objective To investigate the method to isolate and culture hepatic stellate cells ( HSCs) for studying the cellular mechanisms of hepatic frbrosis.Methods HSCs were isolated by nycodenz density gradient centrifugation after the hepatocytes obtained from adult male gebils were digested with pronase, collagenase and DNase, infused via portal vein.The cell viability was determined by trypan blue exclusion test.The purity of HSCs was identified by detectingα-SMA, desmin immunohistochemical staining.Results The yield rate of HSCs was 0.5~1 ×107 per gerbil liver, and the cell viability was more than 90%.The percentage ofα-SMA-positive cells was more than 75%after 3 days primary culture and almost 100% cells were α-SMA and desmin positive in passage culture.Conclusion The successful protocol of primary culture of Mongolian gerbil HSC provide a technical support for research of relevant liver diseases and drug development in the future.

Objective This study is designed to determine whether an association exists between single nucleotide polymorphism (SNP) variant rs2252673 of insulin receptor(INSR) gene and polycystic ovarian syndrome (PCOS) in Han Chinese in order to identify INSR as a genetic susceptibility factor for PCOS.Methods A total of 224 women with PCOS,192 controls and 672 participants consisting of 224 trios (mother,father and offspring with PCOS) were recruited from the Hospital for Reproductive Medicine Affiliated to Shandong University,from July 2007 to April 2013.Genomic DNA was extracted according to the manufacturer' s protocol.SNP rs2252673 of INSR gene was amplified by PCR and then sequenced on an automated sequencer.Moreover,clinical and metabolic features of the patients with PCOS were compared according to the genotypes.The subjects were divided into twot groups according to body mass index (BMI),and then the results were compared between two groups.And the transmitted disequilibrium test (TDT) was applied for data analysis.Results (1) There were three kinds of genotype of CC,CG and GG.Genotype frequencies of rs2252673 were 8.0％,38.8％,53.1％ and 14.6％,42.2％,43.2％ in the PCOS group and the control group,respectively.The allele frequencies of C and G were 27.5％,72.5％ and 35.7％,64.3％ in the PCOS group and the control group,respectively.There were statistical differences in genotype frequencies and allele frequencies between two groups (all P＜0.05).(2)No significant differences were observed in the different genotype according to clinical and metabolic characteristics of women with PCOS (P＞0.05).But when merging the genotype CG and GG,carriers of the CG and GG genotypes in women with PCOS were slightly associated with total cholesterol (TC) levels (t=2.072,P=0.048) and lower high-density lipoprotein (HDL) levels (t=2.274,P=0.026).Although statistical significance was not achieved,there was an increased tendency in fasting blood glucose (FBG) and fasting insulin (FINS) levels in CG and GG genotypes in PCOS cases.(3)Between the obesity and the non-obesity with PCOS,there was no statistical significance in the genotype and allele frequencies (x2=0.054,P=0.974; x2=0.022,P=0.883).(4)The results of families based analysis shown that genotype distribution of the SNP rs2252673 was in Hardy-Weinberg equilibrium (P＞0.05).After the TDT,the G allele in SNP rs2252673 was over transmitted in families (transmitted∶ non-transmitted=120∶ 88; x2=4.923,P=0.027).There was a transmitted disequilibrium in rs2252673,which implies the association of INSR and PCOS were independent of population stratification.Conclusions There were a association between the SNP variant rs2252673 of INSR gene and the susceptibility to PCOS in Han Chinese women,which was independently of body mass index.The carrier of G allele frequency of rs2252673 may have higher risk of PCOS.

Objective To evaluate the relationship between cardiomyopathy-associated 5 (CMYAS) gene rs10043986 polymorphism and schizophrenia in Uygur Chinese population.Methods The SNP rs10043986 in CMYA5 gene was genotyped in 325 patients with schizophrenia and 183 normal controls using TaqMan technology.The symptoms of schizophrenia were assessed by positive and negative syndrome scale (PANSS).The association of the loci with schizophrenia,age of onset,clinical symptom was analyzed.Results The allelic and genotypic distributions in rs10043986 between patients with schizophrenia (C,T allele:91.5％,8.5％ ; C/C,C/T,T/T genotypes:83.4％,16.3％,0.3％) and normal controls (C,T allele:96.4％,3.6％ ; C/C,C/T,T/T genotypes:92.9％,7.1％,0) had statistically significance after analysis (x2 =9.038,P=0.003 ; x2 =9.417,P=0.009).Via analysis of stratification by gender and age at onset.The results showed that both allele (x2=11.812,P=0.001) and genotype (x2=12.769,P=0.001) frequency in rs10043986 with patients were significantly different in females,but neither in males (all P＞0.05).Allelic or genotypic distributions between adult cases and controls had statistically significance (x2=8.219,P=0.004; x2=8.379,P=0.015),but there were not significant differences between adolescent cases and controls (all P＞ 0.05).Furthermore,we also notice that the PANSS scores of patients between Genotype C/C and C/T had no statistically significance (allP＞0.05).Conclusion The results reveal that T allele at CMYA5 rs10043986 may be confer risk for susceptibility of female and adult schizophrenia in Uygur Chinese population,and that rs 10043986 polymorphism may not significantly associate with symptoms severity of schizophrenia.

Objective To establish a fluorescence quantitative Taqman-PCR method for rapid and accurate detection of mouse poxvirus.Methods After sequence alignment and comparison, ERPV_027 gene was selected as the primer and probe design gene.Furthermore, the specificity, sensitivity, stability and reproducibility of these primers and probes were detected.Results The detection limitation of this method was 68 copies/μL.Data showed that this method has high specificity, which specifically amplifies mouse poxvirus, with no amplification signal of mouse hepatitis virus, Sendai virus, Salmonella and some other viruses and bacteria.This method also showed good stability and reproducibility. Conclusions This study has successfully established a fluorescence quantitative Taqman-PCR method for detection of mouse poxvirus, with high specificity, sensitivity, good stability and reproducibility, and a broad application potential.

Objective To explore the clinical results of the arthroscopic acromioplasty in the rotator cuff repair.Methods Sixty-five patients (42 males and 23 females) to receive suture bridge repair under the arthroscopy between May 2012 and May 2014 were selected and randomly divided into an experimental group and a control group.The experimental group was given suture bridge repair with acromioplasty,while the control group underwent the suture bridge repair without acromioplasty.The time of operation was recorded.The range of motion (ROM) of the shoulder,the UCLA shoulder scoring system (ULCA) score,American orthopedic surgeon scoring system (ASES) score,the visual analogue scale (VAS) score and Constant score were observed 3 and 12 months after the operation.Results There were no significant differences in age,sex and the affected side between the two groups.Significant differences were found between the two groups in the duration of the operation (t=-18.5,P＜0.05).There were no significant differences in the ROM and the ULCA,ASES,VAS and constant scores of the shoulder 3 and 12 months after the operation.Conclusion No significant differences were found in the ROM and the shoulder function scoring between giving acromioplasty under the arthroscopy or not.However,the operation time of undergoing acromioplasty under the arthroscopy was longer.

Humans ; Severe Acute Respiratory Syndrome ; diagnosis ; therapy