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 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.

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 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.

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 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 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 evaluate the value of detection of interleukin 28B (IL28B) rs12979860 by allele-specific PCR (AS-PCR) for the prediction of antiviral treatment hepatitis C patients.Methods One hundred seventy-four blood samples were random collected from hospitalized patients with hepatitis C,who came from department of infectious diseases,Renmin Hospital of Wuhan University from May 2011 to May 2012.Two pairs of specific primers were designed for rs12979860 gene polymorphisms,and one mutated base was introduced to the second or third site of the end of 3' with the reverse primer.rs12979860 gene polymorphism of 30 cases with hepatitis C was detected by AS-PCR,and gene sequencing was further used to verify the consistency of the two methods in parallel.Then,the frequency distribution of different rs12979860 genotypes with 174 cases were analyzed by the AS-PCR method in the population.Results The genotype CC,CT or TT of rs12979860 with 30 cases could be well identified by both AS-PCR and gene sequencing,and the coincidence rate was 100％ (x2 =60.0,P ＜ 0.01).Compared to gene sequencing,both of the sensitivity and specificity of AS-PCR were 100％.Compared to the control (CC genotype),TT genotype detection sensitivity by AS-PCR was 10-5,while sequencing sensitivity was 2 × 10-1.rs12979860 polymorphism in the TT,CC and CT genotype distribution in the Chinese population frequencies were 3.45％ (6/174),13.2％ (23/174) and 83.3％ (145/174),respectively.Conclusion AS-PCR can quickly,accurate,reliable,economic and efficiently detect IL28B rs12979860 gene polymorphism of hepatitis C in patients,which could predict the effect of antiviral therapy on patients with hepatitis C.

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.