Objective To detect the genotype of hepatitis C virus(HCV)in chronic hepatitis C(CHC)infection patients using gene sequence method and observe the distributive characteristic of HCV genotype in Hubei.Methods A total of 447 HCV-RNA-positive plasma samples were collected from chronic hepatitis C patients in Infectious Diseases Department of Renmin Hospital of Wuhan University from February 2013 to July 2015.Then NS5B region gene sequence of HCV genome were de-tected by Sanger sequencing method and compared with HCV genotype in NCBI genebank database for analyzing HCV geno-type.Results A total of 11 kinds of genotypes were detected,including genotypes 1a,1b,2a,3a,3b,6a,6b,1b/2a,1b/2k,6a/1b and 6d/6k,respectively.Detection cases of various genotypes were respectively 7 cases (1.57%),325 cases (72.71%),67 cases (14.99%),7 cases (1.57%),20 cases (4.47%),14 cases (3.13%),2 cases (0.45%),2 cases (0.45%),1 case (0.22%),1 case (0.22%)and 1 case (0.22%).Conclusion Genotype 1b was the major type of HCV-RNA genotype,fol-lowed by 2a,also other genotypes existed,which prompted that the prevalence of HCV genotype was diversity in Hubei.

ObjectiveTo evaluate the effect of surgical treatment of distal tibial facture with minimally invasive perutaneous locking compression plate osteosynthesis.Methods38 patients with distal tibial fracture were treated with minimally invasive percutaneous locking compression plate(LCP) osteosynthesis.According to AO classification,twelve patients belonged to type A,fifteen type B,and eleven type C.ResultsAll patients were followed up for 6 to 19 months,with an average time of 9 months.The X-ray pictures showed that one-stage union was achieved in the patients,with a mean healing time of 16 weeks.No complications such as infection,non-union were found.The clinical outcomes were evaluated according to Mazur ankle joint function scoring.Twenty-nine were rated as excellent,six as good and three as fair,with the good-excellent rate of 92.1％.ConclusionMinimally invasive peteosynthesis is a kind of a good fixation in treatment of accelerating bone and soft tissue healing.

Objective To evaluate the applied of selected intraoperative cholangiography(SIOCG) during cholecystectomy.Methods The SIOCG data of 39 patients who had indications for exploration of common bile duct were analysed retrospectively.Results Among 39 cases with SIOCG,abnormal findings were detected in 19 cases(48.72%),in which choledocholith in 16 cases;stenosis of common bile duct at the third part in 3 cases.Positive findings in exploration of common bile duct were 14 cases(73.68%),in which,choledocholith were 12 cases inflammatory stenosis 1 case and tumor of common bile duct at the third part 1 case.Negative patients in exploration of common bile duct were 5 cases(26.32%).Normal imaging in 20 patients who did not underwent exploration of common bile duct had no disorders in follow-up.Conclusion SIOCG is safe,convenient,and useful in preventing the remnant stones and biliary tract damage and improving the quality of cholecystectomy.

BACKGROUND:Ligustrazine hydrochloride which promotes nerve repair can be applied to the treatment of nervous system injury. OBJECTIVE:To investigate the effect of ligustrazine hydrochloride combined with bone marrow mesenchymal stem cels transplantation on electrophysiological property and hindlimb function of rats with spinal cord injury. METHODS:T9 spinal cord transection injury models were made in rats using Alen’s method, and then rat models were randomized into three groups: rats in control group received tail vein injection of culture solution; rats in cel transplantation group underwent bone marrow mesenchymal stem cel transplantationvia the tail vein; rats in combined group were subjected to the tail vein injection of ligustrazine hydrochloride and bone marrow mesenchymal stem cels that lasted for 4 hours. At 1, 2, 4, 6, 8 weeks after modeling, Basso, Beattie and Bresnahan scores and modified Tarlov scores were used to detect the motor function of rats. At 72 hours after modeling, RT-PCR method was used to detect the expression of Bcl-2 and basic fibroblast growth factor around the injured region. At 4 weeks after modeling, somatosensory and motor evoked potentials were measured for evaluation of neurophysiological recovery. At 8 weeks after modeling, horseradish peroxidase tracer was used to assess the regeneration of rat spinal cord nerve fibers; PKH-26 labeling was used to observe the survival and migration of transplanted cels. RESULTS AND CONCLUSION:At 1, 2, 4, 6, 8 weeks after modeling, Basso, Beattie and Bresnahan scores and modified Tarlov scores were significantly higher in the combined group than the cel transplantation folowed by the control group (P < 0.05). At 72 hours after modeling, the expression of Bcl-2 and basic fibroblast growth factor around the injured region was significantly higher in the combined group than the cel transplantation group and control group (P < 0.05). At 4 weeks after modeling, the latencies of somatosensory and motor evoked potentials were ranked as folows: combined group < cel transplantation group < control group (P < 0.05); the amplitudes of somatosensory and motor evoked potentials were ranked as folows: combined group > cel transplantation group > control group (P < 0.05). At 8 weeks after modeling, horseradish peroxidase-labeled pyramidal cels in the cel transplantation group and combined group showed apparent crossing signs; the number of PKH-26-positive cels and horseradish peroxidase-positive cels was the most in the combined group folowed by the cel transplantation group, and was the least in the control group (P < 0.05). These findings indicate that ligustrazine hydrochloride combined with bone marrow mesenchymal stem cels transplantation can facilitate nerve cel regeneration, promote the expression of Bcl-2 and basic fibroblast growth factor, and improve motor function in rats after spinal cord injury. Cite this article:Wu XM, Gao WS, Wang J, Cai HY.Neuroprotection of ligustrazine hydrochloride combined with bone marrow mesenchymal stem cel transplantation in rats with spinal cord injury. Zhongguo Zuzhi Gongcheng Yanjiu. 2016;20(1):95-101.

Objective To explore the mutation spectrum of breast and ovarian cancer susceptibility gene 1/2 (BRCA1/2) which was related to breast cancer in female residents of Wuhan,and assess the relation of gene mutation and risk of suffering breast cancer.Methods 128 cases of female individuals,including 58 cases of breast cancer after surgery,70 cases of benign breast disease,and 50 femal hcalthy volunteers were selected by simple randon sampling from Department of Breast Surgery of Renmin Hospital of Wuhan University.BRCA1/2 gene was sequenced and compared with the standard template sequence to explore the possible mutations.Results In the breast cancer group,mutation emerged in 11 cases and the mutation rate was 19.0％ (11/58),including 8 cases of the BRCA1 gene mutations (3 cases of 185 del AG,5 case of 5382 ins C) and 3 cases of the BRCA2 gene mutations (2 cases of 6174 del T,1 case of C5773T) ; in the benign breast disease group,mutation emerged in 5 cases,the mutation rate was 7.1％(5/70),including 4 cases of the BRCA1 gene mutations (1 case of 185 del AG,3 cases of 5382 ins C),and 1 case of the BRCA2 gene mutation (6174 del T).There was no mutation detected in healthy control group.The mutation rate of the breast cancer group was significantly higher than that of benign breast disease group and healthy control group (x2 =4.05,10.56,P ＜ 0.05); However,there was no significant difference between benign breast disease group and healthy control group (x2 =3.73,P ＞0.05).Conclusions The mutation of BRCA1 gene (185 del AG,5382 ins C) and BRCA2 gene (6174 del T,C5773T) is in the presence of female residents in Wuhan.Furthermore,the mutation in BRCA1/2 gene increases the risk of breast cancer.

Objective To evaluate operative methods and the clinical effect for comminuted calcaneal fractures.Methods 21 cases(24 feet)of comminuted calcaneal fractures were treated by one stage bone grafting plus open reduction and internal fixation with plastic calcaneus titanium plate.The fractures were classified according to the Sanders system into Sanders Ⅱ,Ⅲ and Ⅳ subgroups.Results All cases were followed up for 9～20 months(14months in average).All cases recovered with good healing of fracture.According to Maryland score,the results were excellent in 14 feet,good in 7 feet,fair in 2 feet,and poor in 1 foot.The excellent and good rate was 87.5％.Conclusions By using the method of internal fixation with plastic calcaneus titanium plate,fracture site can be well exposed,which is helpful to recover calcaneus to its normal length,width,height,Gissane angle and Bohler angle.A reliable internal fixation is helpful for early stage functional excise after surgery,which can maximize the recovery of the ankle function.

Objective To evaluate the effects of pretreatment with different doses of phosphocreatine on hepatic ischemia-repeffusion (I/R) injury in rats.Methods.Thirty male Sprague-Dawley rats,weighing 200-250 g,were randomly divided into 5 groups (n =6 each):sham operation group (group S),hepatic I/R group (group I/R),and pretreatment with different doses of phosphocreatine groups (groups P1-3).Hepatic I/R was induced by 90 min occlusion of the hepatic artery and portal vein entering the middle and left lobes of the liver followed by 4 h reperfusion in anesthetized rats.Phosphocreatine 50,150 and 450 mg/kg were injected via the tail vein at 60 min before ischemia in groups P1-3,respectively.In groups S and I/R,the equal volume of normal saline was given instead.Blood samples were taken from the abdominal aorta at 4 h of reperfusion for determination of plasma alanine aminotransferase (ALT),aspartate aminotransferase (AST),TNF-α and IL-1β concentrations.The rats were then sacrificed and the livers were removed for determination of myeloperoxidase (MPO) activity (by ELISA),intercellular adhesion molecule-1 (ICAM-1) expression (by immunohistochemistry),and cell apoptosis (by TUNEL) and for microscopic examination (by electron microscopy).Results The MPO activity in liver tissues,plasma ALT and AST activities,TNF-α and IL-1β concentrations and the number of apoptotic cells were significantly higher in groups I/R and P1-3 than in group S,while lower in groups P1-3 than in group I/R (P ＜ 0.05).The parameters mentioned above were decreasedin turn in groups P1-3 (P ＜ 0.05).Conclusion Phosphocreatine pretreatment can attenuate the hepatic I/R injury in rats in a dose-dependent manner and inhibition of the inflammatory responses is involved in the mechanism.

Objective To evaluated the prevalence of EGFR mutations in Hubei region,to provide reliable experimental basis for reasonable screening TKI beneficiaries in clinic.Methods EGFR mutation of 253 patients diagnosed with NSCLC in Hu-bei region during 2010 to 2015 was detected by PCR-sanger sequencing and ARMS assay,to investigate the sensitivity of dif-ferent methods,to explore the frequency and clinical significance of EGFR mutation in different gender,in different histologi-cal type and different exons.Results 93 of 253 NSCLC patients harbored EGFR mutation,mainly occurred in exon 19 and 21,accounting for 53.76% and 35.38% of the total EGFR mutation rate,respectively.ADx-ARMS method showed higher sensitivity (P=0.001).The mutation detection rate of female NSCLC was significantly higher than that of male NSCLC (P=0.001).The observed incidence of EGFR mutations in patients with adenocarcinomas (38.01%)was the highest in differ-ent histological type,followed by glandular squamous cell carcinoma (30.77%),large cell carcinoma (20%)and squamous cell carcinoma (4.55%).Nonsmoking patients had a higher EGFR mutation frequency (51.6%,81/157)than those with a history of smoking (24%,12/50).Conclusion ARMS assay was more sensitive and more convenient detection method for clinical screening for EGFR-TKI treatment subpopulation.The mutation rate of female NSCLC was significantly higher than that of male NSCLC in Hubei Province.Speculated that the sexual differences in NSCLC with EGFR mutation frequencies were related to hormone levels and smoking status.

Objective To detect UGT1A1 gene promoter polymorphism with gene sequence method and analyze the distribu-tion of UGT1A1*28 and UGT1A1*6 gene polymorphism in Wuhan.Methods A total of 230 samples were collected from tumor patient in Medical Oncology Department of Renmin Hospital of Wuhan University from January 2013 to December 2014,then gene sequence of the target fragments were detected by Sanger sequencing method and analyzed patient UGT1A1 gene promoter polymorphism TA box.Results In this research examination 230 example tumor patient,198 patients (86.1%)were identified with TA6/TA6 genotype,29 patients (12.6%)were identified with TA6/TA7 genotype,3 pa-tients (1.3%)TA7/TA7 variant.Conclusion TA6/TA6 wild-typewas higher frequence in patients with malignant tumour in Wuhan.