Variability of Reverse Transcriptase Gene and S Gene in Lamivudine-treated Chronic Hepatitis B Patients / 病毒学报

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.

Variations of surface, basic core promoter and precore regions in genomes of hepatitis B virus from chronic hepatitis B virus infected patients with positive for both hepatitis B surface antigen and antibody / 中华传染病杂志

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.

Expression and clinical significance of aquaporin 1 and aquaporin 4 in human pulmonary adenocarcinoma / 中国医师进修杂志

ObjectiveTo explore the expression and clinical significance ofaquaporin (AQP) 1 and AQP 4 in human pulmonary adenocarcinoma H1299 cell line.MethodsH1299 cell line in human pulmonary adenocarcinoma(pulmonary adenocarcinoma group) were obtained,the expressions of AQP1 and AQP4 in mRNA level and their locations were determined in H1299 cell line respectively by semiquantitative reverse transcription polymerase chain reaction (RT-PCR) and Western blot analysis.The migration of tumor cells were observed by Matrigel invasion assay.Then normal tissues adjacent of pulmonary adenocarcinoma (above cancer line 3 cm,no tumor cell with pathological proven) were as control group.ResultsThe results of RT-PCR showed that AQP1,AQP4 mRNA was 1.030 ± 0.070 and 1.140 ± 0.190 in conlrol group,which were lower than those in pulmonary adenocarcinoma group (2.021 ± 0.250 and 2.180 ±0.180)(P＜0.05 ).The results of Western blot showed AQP1,AQP4 located on the membrane of H1299 cell.Both AQPI and AQP4 mRNA expressed very high in pulmonary adenocarcinoma group,while expressed very low in control group (P＜0.05).Matrigel invasion assay showed that the invasion was positively related to AQP1,AQP4(r =0.351,P ＜ 0.05 ).ConclusionAQP1,AQP4 significantly over express in H1299 cell line,both of them phy important roles in the growth of tumor tissue and cell migration.

Drug-Resistance of Multi-resistant Pseudomonas aeruginosa / 中华医院感染学杂志

OBJECTIVE To study the mechanism of drug-resistance of multi-resistant Pseudomonas aeruginosa,and provide the guideline for treatment and control of P.aeruginosa infection in hospital.METHODS Fifty strains of multi-resistant P.aeruginosa were selected with K-B susceptibility method.The three-dimensional method was taken to differentiate the various beta-lactamases.The relative drug-resistance gene was detected by polymerase chain reaction(PCR).RESULTS Among 50 strains of multi-resistant P.aeruginosa,there were 2 strains(4%)producing ESBLs,20 strains(40%)producing AmpC beta-lactamases,and 11 strains(22%) producing ESBLs and AmpC beta-lactamases at the same time.There were 8 positive genes in the detected drug-resistance gene,the most common sources of gene were CTX(56%),OprD(60%) and aac(6′)-Ⅱ(60%),respectively.CONCLUSIONS The main beta-lactamases are AmpC beta-lactamases and the main genotype is CTX in the multi-resistant P.aeruginosa cultured in our area.The main course of imipenem-resistance was deletion of outer membrane proteins,and the aminoglycoside modifying enzyme gene and disinfectant-resistance gene in multi-resistant P.aeruginosa are acquired.In order to reduce the drug-resistance strains and control the infection of P.aeruginosa,antibiotics should be used reasonably according to drug susceptibility testing clinically.

Serum HBV-DNA Level in HBsAg Positive Patients Before and after Operation and Their Infectious Risk in Hospital / 中华医院感染学杂志

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

Expression of midkine protein in non-small-cell lung cancer and its clinical significance / 中国癌症杂志

Purpose:To investigate the features of midkine( MK) protein expression and the correlation between MK expression with angiogenes is and prognosis in non-small-cell lung cancer(NSCLC). Methods:Immunohistochemistry analysis for MK and microvessel de nsity(MVD) were performed on 44 samples of tumor tissue. Results:No positivity expression was detected in either the lun g tissue close to tumor or normal lung tissue,but 26 of 44 NSCLC tissue showed a positive reaction to MK immunostain. Significant correlation between MK express ion with MVD enhancement and lymph metastasis in NSCLC was found,and the surviva l terms in NSCLC with MK expression were significantly shorter than those in NSC LC without MK expression.The enhanced MVD was related to lymph metastasis signif icantly,and the survival terms in NSCLC with enhanced MVD were significantly sho rter than those in NSCLC without enhancement. Conclusions:In human non-small-cell lung cancer, MK overexpre ssion at the protein level may very well be closely related to angiogenesis and metastasis. MK protein may be one of the predictors for prognosis in NSCLC.

The expression of midkine(MK) in pancreatic carcinoma and its clinical significance / 中国普通外科杂志

ObjectiveTo investigate the expression of midkine(MK) and its relation with angiogenesis, biological features and prognosis of pancreatic carcinoma(PC).MethodsMK expression and microvessel density(MVD) were determined in 52 cases of human PC with immunohistochemistry and results were compared with pathology.ResultsMean MVD of PC was 64?18 and positive expression of MK was detected in 38 cases (73%). The positive rate of MK was significantly lower in cases of without metastasis and at early clinical stage (stageⅠ～Ⅱ) than that with metastasis and at stage Ⅲ～Ⅳ. MVD was significantly higher in MK-positive PC than in MK-negative PC (P