We wished to undertake molecular characterization of the reverse transcriptase (RT) gene and overlapping surface (S) gene in lamivudine-treated patients with chronic infection with the hepatitis B virus (HBV). Sequencing analyses of the HBV RT/S gene of isolates from 25 chronic hepatitis B (CHB) patients with the YMDD mutation and 30 treatment-naïve CHB patients were undertaken. In patients with the YMDD mutation, rtM2041 was the major type of mutation (20/25, 80%). rtL80I was present in most of the patients with rtM204I (14/20, 70%). rtL180M coexisted with rtM204V (5/5, 100%). Patients with the YMDD mutation had a significantly higher prevalence of mutation of the RT gene than treatment-naïve CHB patients (P < 0.05). Classical primary resistance and secondary/compensatory mutations were detected at only five sites (rtL80, rtV173, rtL180, rtM204, rtM250) in CHB patients with the YMDD mutation. The frequency of nucleos(t)ide analog resistance (NAr) mutation within the RT gene in patients with the YMDD mutation was significantly higher than that in treatment-naïve patients (P < 0.05). Amino-acid mutations within the RT gene were also associated with other types of NAr in patients with the YMDD mutation. The rate of amino-acid variants within the S gene region was significantly higher in patients with the YMDD mutation than that in treatment-naïve patients (P < 0.05). sM133L and sG145R variants were also present in patients with the YMDD mutation. These observations suggest that CHB patients with the YMDD mutation also have NAr mutations related to other NA drugs, which might lead to cross-resistance in CHB patients. Variants present in the S gene region could cause changes in the antigenicity of HBsAg, which could result in a false-negative diagnosis of HBsAg and immune in escape of the HBV.
Adolescent ; Adult ; Antigens, Surface ; genetics ; Antigens, Viral ; genetics ; DNA Mutational Analysis ; Female ; Genetic Variation ; Hepatitis B, Chronic ; drug therapy ; genetics ; Humans ; Lamivudine ; pharmacology ; therapeutic use ; Male ; Middle Aged ; RNA-Directed DNA Polymerase ; genetics ; Young Adult

Background:Gankyrin is an ankyrin repeat oncoprotein overexpressed and involved in the tumorigenesis and progression of various cancers. Aims:To investigate the effect and underlying mechanism of down-regulation of gankyrin expression on proliferation of gastric cancer cells. Methods:Lentivirus vector carrying gankyrin-targeted siRNA was transfected into human gastric cancer cell line MKN28. Cell proliferation,cell cycle distribution and β-catenin/ cyclin D1 signaling pathway was analyzed by MTT assay,flow cytometry and Western blotting,respectively,in gankyrin-silenced MKN28 cells and control cells. Results:The transfection efficiency of lentivirus vector was more than 90% ,and the protein expression of gankyrin in gankyrin siRNA transfected MKN28 cells was significantly repressed( P ﹤ 0. 01). Compared with cells transfected with control lentivirus and cells without transfection,MKN28 cells transfected with gankyrin siRNA showed markedly repressed cell growth after 3-day-culture;the proportion of cells in cell cycle G1 phase was significantly increased,and that in S phase was significantly decreased;down-regulated expression of β-catenin and cyclin D1 was observed(P all ﹤ 0. 01). Conclusions:Down-regulation of gankyrin expression in gastric cancer cells may induce cell cycle G1 phase arrest and inhibit cell proliferation by suppressing β-catenin/ cyclin D1 signaling pathway. Gankyrin might be a promising novel target for targeted therapy of gastric cancer.

Objective To analyze the variations of surface(S) region,basic core promoter (BCP) and precore (preC) regions in genomes of hepatitis B virus (HBV) from patients with coexistence of hepatitis B surface antigen (HBsAg) and hepatitis B surface antibody (anti-HBs).Methods S region,BCP and preC regions in genomes of HBV were amplified and sequenced in 62 HBV-infected patients including 27 HBsAg-positive/anti-HBs-positive patients (double positive group) and 35 HBsAg-positive/ anti-HBs-negative patients (single positive group).The sequencing results and amino acid variants in these regions were analyzed.Difference of means between groups was compared by t test.Sample rate and variation rate were compared by chi-square test.Results One hundred and fifty-six amino acids mutations within the S region were detected in 27 patients of double positive group and 100 mutations in 35 patients of single positive group.The mutation rate in double positive group was significantly higher than those in single positive (2.56％ vs 1.26％,x2 =32.07,P＜0.05).The amino acid variants in double positive group were much higher than those in single positive group within major hydrophilic region (MHR),especially in the first loop area of a-determinant in S region (4.76 ％ vs 1.02 ％,x2 =11.58,P＜0.05).The mutation rate of A1762T/G1764A in BCP in double positive group was significantly higher than those in single positive group (59.3％ vs 28.6％,x2 =5.895,P＜0.05).The mutation rate of A1846T in preC region was higher in double positive group than those in single positive group (40.7％ vs 17.1％,x2-4.265,P＜0.05).The mutation rate of A1762T/G1764A+G1896A in double positive group was also higher than that in single positive group (37.0％ vs 14.3％,x2 =4.302,P＜0.05).Conclusions The mutation rates of S region,especially in the first loop area within a-determinant,BCP and preC regions which are related with hepatocellular carcinoma development in HBsAg and anti-HBs double positive group are higher than those in HBsAg single positive group in chronic HBV infected patients.

OBJECTIVE To investigate the changes in serum HBV-DNA level in HBsAg positive patients before and after operation and their infectious risk in hospital.METHODS HBV markers(HBV-M) in serum was detected in 58 HBsAg positive patients by time-resolved fluoroimmunometric assay before operation.HBV-DNA level in serum of them before operation and at 3rd,and 7th day after operation was detected by real time fluorescent quantitative polymerase chain reaction.We also detected HBV-DNA in gastric drainage juice and abdominal drainage after operation.RESULTS HBV-DNA was detected in 27 of 58 HBsAg positive patients' serum,the positive rate was 46.1%.After operation,serum HBV-DNA was increased remarkably at 3rd and 7th day compared with before operation in these patients respectively(P

Objective:To analyze the independent diagnostic indicators and their diagnostic values for pulmonary tuberculosis complicated with pulmonary embolism.Methods:A total of 34 cases of pulmonary tuberculosis complicated with pulmonary embolism treated in Huzhou Central Hospital from March 2014 to September 2021 were enrolled. And 136 patients with simple pulmonary tuberculosis who were hospitalized during the same period were collected with a ratio of 1∶4 according to the principle of age and gender matching. The general conditions, clinical symptoms, comorbidities and laboratory indicators of the patients were retrospectively analyzed. The univariate analysis was performed using independent samples t test, Mann-Whitney U test and chi-square test. Binary logistic regression was used to analyze the related diagnostic factors for pulmonary embolism in pulmonary tuberculosis patients, and the combined factors were constructed by transforming the model equation, and the receiver operating characteristic (ROC) curve was used to determine the optimal cut-off value and evaluate its diagnostic value. Results:The univariate analysis showed that patients with pulmonary tuberculosis complicated with pulmonary embolism had higher ratio of chest tightness (67.6%(23/34) vs 22.1%(30/136)), syncope (23.5%(8/34) vs 0.7%(1/136)), fever (55.9%(19/34) vs 36.0%(49/136)), hemostatic drug use (100.0%(34/34) vs 13.2%(18/136)), history of venous thrombosis (8.8%(3/34) vs 0.7%(1/136)), atrial fibrillation (11.8%(4/34) vs 2.2%(3/136)) and D-dimer levels (4.090 0(1.035 0, 10.790 0) mg/L vs 0.850 0(0.432 5, 2.145 0) mg/L) than those of simple pulmonary tuberculosis patients, and the differences were all statistically significant ( χ2=26.35, 28.19, 4.47, 96.44, 7.75, 6.30 and Z=-4.65, respectively; all P<0.050). The arterial partial pressure of oxygen (PaO 2)(61.90(52.95, 73.00) mmHg vs 82.00 (75.00, 87.00) mmHg, 1 mmHg=0.133 kPa) and albumin ((28.83±4.98) g/L vs (32.76±5.65) g/L) of patients with pulmonary tuberculosis complicated with pulmonary embolism were lower than those of simple pulmonary tuberculosis patients, and the differences were both statistically significant ( Z=-5.21 and t=3.71, respectively, both P<0.001). Binary regression analysis showed that chest tightness (odds ratio ( OR)=3.494, 95%confidence interval ( CI) 1.208 to 10.100, P=0.021), D-dimer ( OR=1.285, 95% CI 1.079 to 1.530, P=0.005) and PaO 2( OR=0.931, 95% CI 0.895 to 0.970, P=0.001) were the independent diagnostic indicators for pulmonary embolism in pulmonary tuberculosis patients. The areas under the ROC curve of chest tightness, D-dimer, PaO 2, and the combination of the three indicators (the combination factor) were 0.728, 0.758, 0.834, and 0.890, respectively. The optimal cut-off value of the combination factor was -3.1, with the sensitivity of 0.824 and the specificity of 0.824. Conclusions:Chest tightness, increased D-dimer and decreased PaO 2 are independent diagnostic indicators for pulmonary embolism in pulmonary tuberculosis patients. It is recommended to perform pulmonary artery computed tomography angiography promptly when the combination factor is higher than -3.1 to determine whether the patient is complicated by pulmonary embolism.