Tumor markers refer to the specific substances that exist in tumor cells themselves or are secreted by tumor cells. They can reflect the existence and growth of the tumor. The serum tumor biomarkers have been widely applied in tumor detection,but the detection of these markers is based on the tumor antigens,and thus has many inadequacies in tumor screening and diagnosis. In this paper,we reviewed autoantibodies as tumor biomarkers against self-antigens in vivo. This method is to examine the tumor autoantibodies by using the tumor antigens,and its specificity and sensitivity are superior to the traditional examination methods. Using autoantibodies to detect tumors would provide a new method for tumor screening and diagnosis.

Objective To investigate the effect of end to side arterial anastomosis in the transplantations of free tissue flap Methods The end to side arterial anastomosis were used in 47 free flaps of 46 cases in this series The arterial stomas were respectively prepared into the cross section (13/47),the inclined plane angled at 45 degree (26/47) and fish mouth shaped plane (8/47) The interrupture anastomosis and continuous anastomosis following two fixed point sutures,and the continuous anastomosis following four fixed point sutures were respectively used in 28,10 and 9 arterial anastomotic stomas Results There were not postoperative embolism in all end to side anastomotic stomas in this series During the operations the embolisms of the cutaneous branch caused by a detached mini thrombus from the anastomotic stomas were occurred in 2 cases,and the arterial crisis were eliminated by reversely milking out the detached mini thrombus All flaps survived except one due to venous embolism Conclusions It is a effective and safe method that the end to side arterial anastomosis are used in the transplantation of free flap

Objective The purpose is to report the clinical result of repairing defection in hand surgery by using both the vascular tissue flap and external fixators. Methods The vascular tissue flap including bone flap and joint flap to repair the complex defection (soft tissue, bones and joints) in hand and forearm were applied in 9 cases. At the same time in surgery,injured bones and joints were fixed by A-O and mini external fixators in order to keep the non-injured joint actively and accelerate the cure of bones. Result All 9 cases were applied successfully with the all transplanted tissue survived. Pulling about the external fixator about 10- 12 weeks after surgery showed bone union and the f unction of hand recovered satisfactorily' Conclusion it is a reliable method for repairing the complex tissue defection in hand and forearm by combining external fixation of traumarology and microsugery technique. And it is also characterized by a minor injury in transplanted bones,early exercise for joints,flap circulation observed conveniently,and nursing easily.

With the development of molecular diagnostic techniques, the study for the etiology of leukemia has been entering the era of the molecular biology.Molecular techniques for leukemia diagnosis, prognosis and individualized therapy are used widely, becoming one of the necessary routine tests for patients with leukemia. So far, molecular techniques for leukemiaincluding cytogenetic diagnosis, molecular genetics, molecular diagnostics based on PCR, mutation detection, as well as a variety of next-generation sequencing, have played an important role in the diagnosis of leukemia, minimal residual disease monitoring, prognosis and targeted therapy.

Objective Investigating the expression of TLR3 and TLR7 mRNA in peripheral blood mononuclear cells (PBMC) of patients with chronic hepatitis C infection (CHC),to explore the effects of gender and age on Toll-like receptor expres-sion.Methods Peripheral blood was collected from 1 1 5 patients of chronic hepatitis C infection,1 1 3 healthy individuals.Ex-pression levels of TLR3 and TLR7 mRNA were detected by real-time quantitative PCR.Results The expression of TLR3 and TLR7 mRNA were significant difference between patients with CHC infection and healthy individuals (t=38.73,6.16, P<0.05),respectively.The expression of TLR3 and TLR7 mRNA were significant difference between premenopausal fe-males with CHC infection,postmenopausal females and young males of CHC infection (t=61.210,6.464,P<0.05;t=24.166,26.266,P<0.05),respectively.The expression of TLR3 and TLR7 mRNA were significant differences between old males and young males of chronic hepatitis CHC infection (t=86.349,19.583,P<0.05).The expression of TLR3 mRNA was significant differences between old males and postmenopausal females of CHC infection (t=122.941,P<0.05).There was no correlation between the expressions of TLR3 and TLR7 and HCV-RNA load of CHC patents of Gender and age dis-crepancies (|r|<0.40,P>0.05).The HCV-RNA load of premenopausal females was significant lower than young males and old females (t=3.49,2.51,P<0.05),the load of old males was lower than old females (t=2.35,P<0.05),however, there was no significant differences between old males and old females (t=1.20,P<0.05).Conclusion Gender and age dis-crepancies have a relationship with the expression of Toll-like receptors of patients with CHC infection,which may provide a theoretical basis and a new method for CHC.

Objective To investigate the association of rs10004195 single nucleotide polymorphisms in Toll like re-ceptor 10(TLR 10) gene with helicobacter pylori infection and associated diseases. Methods A total of 652 pa-tients who has been examined by gastroscopy were obtained, and then peripheral blood samples were collected from all the patients. Immune colloidal gold method was used to test the serological Hp antibody of all the patients. The TLR10 gene rs10004195 polymorphisms were examined by direct DNA sequencing of the PCR products. Results The frequencies of AA,TT and AT genotype on TLR10 rs10004195 were 30. 98%, 20. 71%, 48. 31%;there was significant difference between Hp antibody positive group and Hp antibody negative group in the TT frequencies of TLR10 rs10004195 ( P<0. 05 ) . No significant difference between controls and Hp associated diseases groups in Hp antibody positive group or in Hp antibody negative group were observed. Conclusion There was correlation be-tween the TLR10 rs10004195 loci genotype and the risk of Hp infection, but no correlation with Hp associated dis-eases.

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 analyze the association of the single nucleotide polymorphism (SNP) of Toll-like receptor 7 (TLR7) and Toll-like receptor 9 (TLR9) with chronic hepatitis C virus (HCV)infection.Methods A total of 150 patients with chronic hepatitis C (CHC) admitted to Renmin Hospital of Wuhan University from January 2011 to May 2012 and 168 healthy controls were enrolled in the study.The genotypes of TLR7 IVS2-151 (rs179009) were detected by Sanger sequencing,and the genotypes of TLR9 T-1486C (rs187084) were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP).SPSS 15.0 was used for statistical analysis,and goodness-of-fit test for HardyWeinberg equilibrium was also performed.Results The frequency of TLR7 IVS2-151G was higher in malepatients with CHC than that in male controls (41.4％ vs.21.6％,x2 =7.250,P =0.007,OR =0.389,95％ CI:0.194-0.781) ; however the female CHC patients had a higher frequency of TLR7 IVS2-151A than the female controls (76.9％ vs.63.1％,x2 =7.202,P =0.007,OR =1.942,95％ CI:1.192-3.164).No significant difference in the distribution of TLR9 T-1486C (rs187084) gene SNP was observed betweenCHC and control groups (P ＞0.05).Conclusion TLR7 IVS2-151 (rs179009) is correlated with HCV infection,which may be involved in the pathogenesis of CHC.

Objective To explore the accuracy and clinical application of Pyrosequencing for detection of drug resistant relevant mutation in the polymerase gene of Hepatitis B virus (HBV).Methods Compared with Sanger sequencing,the accuracy and sensitivity of Pyrosequencing were assessed.Pyrosequencing was used to determine the serum of 1164 patients with chronic Hepatitis B and its results were analyzed.Results The sensitivity of Pyrosequencing was 1 × 103 KIU/L,the same as Sanger sequencing.But Pyrosequencing was more stable and specific than Sanger sequencing to detect low rate of mutation with over 10％ mutation.The mutations couldn't be detected in 248 patients with chronic Hepatitis B,but positive results occurred in 919 patients,among them,61.5 ％ without mutation,38.5 ％ with mutation (including 47.5％ of the single locus mutation and 52.5％ of the combined mutation).Other sites had different degree of mutations except the sites of rtI169T and rtA194T; the main mutation sites in patients with chronic Hepatitis B were rtM204V/I (32.1％),rtL180M (19.8％),rtA181V/T (6.7％) and rtN236T (5.3％),in which fractional mutation had high ratio in the sites of rtA181V/T (6.7％) and rtN236T (5.3％).Conclusions Pyrosequencing has good sensitivity,specificity and accuracy and can early detect the drug resistant relevant mutation in the polymerase gene of HBV,which is suitable for monitoring patients with chronic Hepatitis B for the treatment of nucleoside analogues (acid).The different sites and frequencies of mutations may be associated with different habits of doctors for using different anti-HBV drugs.