0.05). The sequence of five clones from one serum which was identified precore mutation by mPCR-RFLP were all A1896 mutant strains.Another serum identified mixture infection by mPCR-RFLP , one clone was A1896 mutant strain and four were G1896 wild strains.The results of mPCR-RFLP were verified by cloning.Conclusions Comparison with sequencing, the mPCR-RFLP method is simple,accurate and can be used in large-scale surveys and clinical research.

Objective To study association of hepatitis B virus(HBV) precore (pre c)/basic core promoter(BCP) mutations with the genotype or the progression of liver disease. Methods The serum samples from 148 patients with HBV-relative diseases were collected, including 31 asymptomatic carriers, 32 with chronic hepatitis B (CHB), 40 with liver cirrhosis(LC) and 45 with hepatocellular carcinoma(HCC). The genes covering HBV pre c and BCP were amplified by nested polymerase chain reaction (nPCR). The PCR products were subjected to direct sequencing and the mutations in pre c 1896 and BCP 1762/1764 were determined by sequence analysis. HBV genotypes were also detected in the sera by restriction fragment length polymorphism based on S-gene PCR products. Results Of 148 serum samples of HBV, 128 were successfully genotyped and sequenced. There were 60 genotype B and 68 genotype C. The mutation in pre c (A1896) was significantly higher in genotype B than in genotype C (48.3% vs 29.34%, P

ObjectiveTo observe the efficacy of L-carnitine in prevention and treatment of toxicity of LFP chemotherapy with gastrointestinal cancer. Methods60 cases of gastrointestinal cancer were divided into 2 groups according to the admission date. The treatment groups received LFP chemotherapy and L-carnitine, while the control group received LFP chemotherapy alone.Both groups received 3 cycles chemotherapy. The gastrointestinal toxicity, neurotoxicity, hematologic toxicity and physical state were compared.ResultsThere were 12 cases of peripheral neurotoxicity in treatment group,the incidence rate was 40.0 ％;but in control group there were 21 patients, and the incidence rate was 70.0 ％. There was a significant difference between the two groups(x2=5.4545,P =0.0195). Anemia in the treatment group was 56.7 ％ (17/30); in the control group the rate was 86.7 ％ (26/30).There was a significant difference between the two groups (x2 =6.698,P =0.0351).After chemotherapy,in the treatment group,there were 4 cases with increasing Karnofsky score ≥ 10,19 patients whose Karnofsky score did not change,and 7 cases with reduced Karnofsky score ≥ 10; in the control group,there was 1 case with increasing Karnofsky score ≥ 10,13 cases with Karnofsky score no changing,and 16 cases with reduced Karnofsky score ≥ 10.The changes in physical state of two groups had a significant difference(x2 =5.711,P =0.0169).The gastrointestinal toxicity,thrombocytopenia,and neutropenia of two groups had no significant difference.ConclusionL-carnitine can reduce peripheral neurotoxicity and hematologic toxicity,improve the patient's physical state in patients received LFP chemotherapy.

OBJECTIVETo investigate the prevalence of transfusion-transmitted virus (TTV) infection and analyze partial nucleotide sequence of TTV isolated from Guizhou area.

METHODSA nested polymerase chain reaction (nPCR) assay with two-set of primers deduced from the first read frame of TTV genome was established, the prevalence of TTV DNA were detected in 395 sera collected from different population in Guizhou area. The partial genes of TTV DNA fragments amplified from two liver disease patients and one hemodialysis patient were analysed and then compared with gene from Japan strain.

RESULTSThe positive rates of TTV DNA in 62 normal adults, 37 blood donors, 50 hemodialysis patients, 107 intravenous drug users (IVDUs), 139 patients with liver diseases were 6.45% (4/62), 8.10%(3/37), 26.00% (13/50), 25.23% (27/107) and 16.54% (23/139), respectively. The homology of TTV DNA sequence was from 99% to 100% within three Guizhou strains. As compared with published sequence, the nucleotide homology is 98% between all three Guizhou strains and Japan strain.

CONCLUSIONSThere are TT virus infection in Guizhou area, the infection rates were higher in hemodialysis patients and IVDUs. The nucleotide homology was high between Guizhou strain and Japan strain.

Adolescent ; Adult ; Aged ; Blood Donors ; China ; epidemiology ; DNA Virus Infections ; epidemiology ; virology ; DNA, Viral ; blood ; Female ; Humans ; Male ; Middle Aged ; Polymerase Chain Reaction ; Sequence Analysis, DNA ; Torque teno virus ; genetics ; isolation & purification

Objective:To provide a potential platform for transferring specific antigen against fish bacterial diseases based on attenuated Lm (EGDe-ΔactA/ΔinlB).Methods: Attenuated Lm (EGDe-ΔactA/ΔinlB) was used to express outer membrane protein K (Ompk),a conserved and effective vaccine candidate in vibrio.The identification of recombinant strains and detection of antigen genes were operated with PCR and RT-PCR,respectively.Results: The results of PCR showed that Lm-Ompk (L-O),Lm-Lmo0576-Ompk (L-L-O) and Lm-P-Ompk (L-P-O) were constructed successfully.The identity of foreign gene was 100％ compared with sequence of NCBI.The analysis of transcription showed that the expressions of Ompk in L-O,L-L-O and L-P-O were significant (P<0.001).Moreover,the expression of Ompk in the condition of antibiotic was higher than that in the BHI without antibiotic (P<0.05).Conclusion: Lm-Ompk (L-O),Lm-Lmo0576-Ompk (L-L-O) and Lm-P-Ompk (L-P-O) were constructed successfully.

OBJECTIVETo assess the association of CYP2C19 gene polymorphisms with the incidence of ischemic stroke among patients receiving clopidogrel therapy following coronary stenting for coronary artery disease.

METHODSClinical data of patients receiving clopidogrel therapy after coronary stenting were retrospectively studied. For a case-control study, 137 patients with acute cerebral infarction and 122 non-stroke patients were selected. Based on the variants of the CYP2C19 gene detected by a DNA microarray assay, the patients were further divided into the wild-type group(CYP2C19*1/*1) and mutant group(defined by the presence of at least one loss-of-function allele, including CYP2C19*1/*2, CYP2C19*1/*3, CYP2C19*2/*2, CYP2C19*2/*3 and CYP2C19*3/*3). The incidences of ischemic stroke in the two groups were compared through a chi-square analysis. The influence of CYP2C19 gene polymorphisms and clopidogrel therapy on the incidence of ischemic stroke was analyzed through multivariable logistic regression.

RESULTSA total of 259 patients were enrolled. The case and control groups showed no difference in terms of gender and age. There were 123 cases (47.5%) in the CYP2C19 wild-type group and 136 cases (52.5%) in the mutant group. The incidence of ischemic stroke of mutant group was significantly higher than that of wild-type group (59.9% vs. 44.3%, X2=6.398, P=0.042). Multivariate analysis revealed that loss-of-function polymorphisms of the CYP2C19 gene carried a 1.13 times greater risk for ischemic stroke compared to wild-type genotype (OR=2.13, 95%CI: 1.23-3.71).

CONCLUSIONThe efficacy of clopidogrel for the prevention of ischemic stroke in post-coronary stent patients may be reduced by the insufficiency of the CYP2C19 gene. The dosage of clopidogrel therapy should be adjusted based on its polymorphisms.

Brain Ischemia ; prevention & control ; Cytochrome P-450 CYP2C19 ; genetics ; Genotype ; Humans ; Percutaneous Coronary Intervention ; adverse effects ; Platelet Aggregation Inhibitors ; therapeutic use ; Polymorphism, Genetic ; Stents ; adverse effects ; Stroke ; prevention & control ; Ticlopidine ; analogs & derivatives ; therapeutic use