Correlation of the left coronary artery morphology and occurrence of acute myocardial infarction with left main lesion and in-hospital mortality / 中国介入心脏病学杂志

Objective To discuss the correlation of the left coronary artery morphology and occurrence of acute myocardial infarction (AMI) with left main lesion and in-hospital mortality.Methods This was a retrospective study of clinical information and angiographic data of 176 patients admitted from January 2010 to April 2015 with left main lesion in the First Affiliated Hospital of Harbin Medical University.The patients were divided into left main acute myocardial infarction group and left main non-acute myocardial infarction group,and then the left main AMI group was further divided into the survival group and the deceased group according to the situation of in-hospital death.QCA software was used to measure the anatomical morphology of left coronary artery.The anatomical morphology of left coronary artery between the two groups was compared,and univariate analysis and logistic regression analysis were applied to analyze the factors of AMI occurrence and in-hospital mortality in AMI patients with left main lesion.Results The angle between the left anterior descending artery (LAD) and the left circumflex artery (LCX) of the LM-AMI group was smaller than the non-acute myocardial infarction group (P＜0.05),but there was no significant difference in the diameter and length of left main coronary artery (LM),the diameter of LAD and LCX,the angle between LM and LAD and the angle between LM and LCX between the two groups (P＞0.05).The risk of LM-AMI in patients with the angle between LAD and LCX of ＜ 79.43°is 3.6 times right than patients with the angle between 79.43°～ 108.73°.Between the survival group and the deceased group of AMI patients with left main lesion,there were no significant differences in the diameter and length of LM,the diameter of LAD and LCX,the angle between LM and LAD,the angle between LM and LCX and the angle between LAD and LCX (P＞0.05).Conclusions There was a correlation between the angle between LAD and LCX with the occurrence of LM-AMI in patients with left main lesion.The angle between LAD and LCX of ＜ 79.43° is the independent risk factor of LM-AMI in patients with left main lesion,but there was no correlation between the left coronary artery morphology and in-hospital mortality of patients with left main AMI.

Expression of RRM1 and ERCC1 genes in tumor tissues and peripheral blood lymphocytes of advanced non-small cell lung cancer / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the expression of RRM1 and ERCC1 genes in tumor tissues and peripheral blood lymphocytes of advanced non-small cell lung cancer (NSCLC).</p><p><b>METHODS</b>Tissue and peripheral blood samples were collected from 49 advanced NSCLC patients treated with gemcitabine plus carboplatin. The expressions of RRM1 and ERCC1 mRNA in tumor tissue and peripheral lymphocytes were detected by real-time fluorescent quantitative PCR. The relationship of gene expression with clinical characteristics,chemotherapy response and prognosis was analyzed.</p><p><b>RESULTS</b>The RRM1 expression in tumor tissues was positively correlated with that in peripheral blood lymphocytes,while no significant correlation was observed between ERCC1 expression in tumor tissues and that in peripheral blood (rs=0.332 and 0.258; P=0.020 and 0.073, respectively). The expression of RRM1 and ERCC1 in tumor tissues peripheral lymphocytes was synchronous (rs=0.634 and 0.351; P<0.001 and 0.013, respectively). There was no significant correlation of gene expression with gender, age, smoking status, performance status, clinical stages and histological types of patients (P>0.05). Significant difference was found in response rate to chemotherapy (P<0.05,P<0.01,P<0.05),median survival time (P<0.05,P<0.01,P<0.05) and 1-year survival rate (P<0.01,<0.05,P<0.05) between patients with low RRM1 and ERCC1 expression levels in tumor tissues or low RRM1 expression levels in peripheral blood and those with high RRM1 and ERCC1 expression levels. The patients with low ERCC1 expression levels in tumor tissues gained higher 2-year survival rate (P<0.05). There was no correlation of the expression of ERCC1 in peripheral blood with the response to chemotherapy and prognosis (P>0.05).</p><p><b>CONCLUSION</b>The expression of RRMI and ERCC1 genes in tumor tissues and RRM1 in peripheral blood lymphocytes is closely correlated with the response to chemotherapy and prognosis of patients with advanced NSCLC treated with gemcitabine plus carboplatin.</p>

Detection of RRM1, ERCC1 and BRCA1 gene expression in non-small cell lung cancer tissues and peripheral blood by SYBR real-time fluorescent quantitative PCR / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To develop a method for the detection of RRM1, ERCC1 and BRCA1 gene expression by SYBR real-time fluorescent quantitative PCR in non-small cell lung cancer tissues and peripheral blood.</p><p><b>METHODS</b>The plasmid standard of RRM1, ERCC1, BRCA1 and β-actin genes was constructed. SYBR real-time PCR was performed, and the standard curve was established. The expressions of RRM1, ERCC1 and BRCA1 mRNA in non-small cell lung cancer tissues and peripheral blood were detected.</p><p><b>RESULT</b>The standard curve presented linearity. The liquate curves of standard gene were all single apex, indicating that a good specificity was obtained.</p><p><b>CONCLUSION</b>The developed SYBR real-time fluorescent quantitative PCR has advantage of convenient operation, low cost, good specificity and high veracity.</p>

Solid lipid nanoparticles loading adefovir dipivoxil for antiviral therapy / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Herein, solid lipid nanoparticles (SLN) were proposed as a new drug delivery system for adefovir dipivoxil (ADV). The octadecylamine-fluorescein isothiocynate (ODA-FITC) was synthesized and used as a fluorescence maker to be incorporated into SLN to investigate the time-dependent cellular uptake of SLN by HepG2.2.15. The SLN of monostearin with ODA-FITC or ADV were prepared by solvent diffusion method in an aqueous system. About 15 wt% drug entrapment efficiency (EE) and 3 wt% drug loading (DL) could be reached in SLN loading ADV. Comparing with free ADV, the inhibitory effects of ADV loaded in SLN on hepatitis B surface antigen (HBsAg), hepatitis B e antigen (HBeAg) and hepatitis B virus (HBV) DNA levels in vitro were significantly enhanced.

Character of HBV (hepatitis B virus) polymerase gene rtM204V/I and rtL180M mutation in patients with lamivudine resistance / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

<p><b>OBJECTIVES</b>To investigate the relationship between HBV (hepatitis B virus) polymerase gene 180 and 204 sites mutation and lamivudine resistance.</p><p><b>METHODS</b>One hundred forty-one patients with lamivudine resistance after lamivudine treatment and 60 chronic hepatitis B patients without lamivudine treatment were enrolled in this study. The serum HBV DNA mutation was analyzed by sequence detection via polymerase chain reaction (PCR). The sequences of the same patient were analyzed before and after lamivudine treatment.</p><p><b>RESULTS</b>One hundred and nine lamivudine resistance patients had HBV YMDD (tyrosine-methionine-aspartate-aspartate) mutation. Among them, 45 patients had rtL180M/M204V mutation (41.28%), 28 patients had rtL180M/M204I mutation (25.70%) and 36 patients had rtM204I mutation (33.02%). There were 6 patients with rtL180M mutation in 32 lamivudine resistance patients. Sixty chronic hepatitis patients without lamivudine treatment had no mutations.</p><p><b>CONCLUSIONS</b>HBV mutations, which play an important role in lamivudine resistance usually locate at polymerase gene 204 site; 180 site mutation was also observed in these patients. Evaluation of the anti-virus therapy by surveillance of the two sites mutations is of importance.</p>

Effect of fragile histidine triad gene on apoptosis of MGC-803 gastric cancer cells / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the effect of fragile histidine triad (FHIT) gene on the apoptosis of gastric cancer cells.</p><p><b>METHODS</b>FHIT gene was transfected into gastric cancer cell line MGC-803 by liposome. Western blot and immunohistochemical assay were employed to detect the expression of FHIT. Apoptosis and cell cycle were analyzed by flow cytometry (FCM). Morphological changes were observed by light and electron microscope.</p><p><b>RESULTS</b>Stable expression of FHIT protein in transfected MGC-803 cells was obtained. The rates of G0/G1 phase and apoptosis cells were significantly higher in FHIT-transfected MGC-803 cells than those in control cells (64.4%compared with 49.5%and 32.0%compared with 12.2%, respectively).</p><p><b>CONCLUSION</b>The results indicate that FHIT gene might be involved in the apoptosis of gastric cancer cells.</p>

Development of a system for quick screening of efficient HBx-siRNA / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To develop a system for quick screening of efficient siRNA targeted HBx mRNA.</p><p><b>METHODS</b>Using recombination DNA technique, the fusion expression plasmid of HBx and EGFP was constructed, and siRNA expression cassettes (SECs) containing U6+1, H1 or tRNA(Val )promoter were prepared via one-step overlapping extension PCR. By co-transfection with recombinant plasmid and SECs into AD293 cell, the inhibition effects on the transient expression of HBx-EGFP fusion protein were analyzed by FACS and semi-quantitated RT-PCR analysis.</p><p><b>RESULT</b>(1)HBx-EGFP fusion protein expression plasmid pHBx-EGFP was constructed successfully, which expressed green fluorescence in cell mainly located at plasma or the periphery of nucleus in granules. (2) Co-transfection with recombinant plasmid and SECs into AD293 cells resulted in inhibition of HBx-EGFP expression. SEC-siHBx388 showed significant inhibition effect on HBx-EGFP expression compared with SEC-siHBx271, indicating that siHBx388 is effective siRNA site and could be screened out with our screening system. In addition,the results of that U6+1-, tRNA(Val) and H1-siHBx388 reduced HBx-EGFP expression by 21.7%, 12.9% and 12.4% of control respectively indicated that both tRNAVal and H1 promoter was high efficient in driving effect of siHBx388.</p><p><b>CONCLUSION</b>Combination of the HBx expression carrying reporter gene and PCR-based multi promoter SECs may develop a useful system to be applied in identification of optimal HBx- siRNA and its matching promoter.</p>

Establish a screening system for selection of mRNA target sites for HBsAg to construct siRNA with shRNA / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To find some effective short interfering RNA's sites targeting HBV surface gene sequence using shRNA expression vectors.</p><p><b>METHODS</b>Four shRNA expression vectors targeting HBV surface gene sequence were constructed based on pAVU6 + 27 vector, and cotransfected into AD293 cells with HBs-EGFP fusion gene plasmid. The changes of HBs-EGFP image were detected by FACS and microscopy. The HBs-EGFP mRNA expression was evaluated by RT-PCR.</p><p><b>RESULTS</b>Four shRNA expression vectors and HBs-EGFP fusion gene plasmid were successfully constructed. pAVU6 + 4sh579 vector inhibited the HBs-EGFP expression by 69.8% in AD293 and suppressed the HBs-EGFP mRNA expression by 74.6%.</p><p><b>CONCLUSIONS</b>The results showed that the 579 site of HBV surface gene sequence was an effective target and pAVU6 + 4sh579 vector could suppress the HBs-EGFP expression in AD293 cells</p>

Expression of IFN-gamma and its receptor alpha in the peripheral blood of patients with chronic hepatitis C / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>It has been known that intra-cellular immunity is important for defense against viral infections and this function lies with interferon gamma (INF-gamma). Here we evaluated the role of IFN-gamma system in the pathogenesis of chronic hepatitis C (CHC).</p><p><b>METHODS</b>The levels of interferon gamma receptor alpha (IFNGR alpha) on the peripheral lymphocyte membrane were assayed with flow cytometry. The plasma concentrations of the cytokines IFN-gamma and IL-10 in CHC patients and normal controls were assayed by enzyme-linked-immunosorbent assay (ELISA). The samples were collected randomly from Xinjiang Autonomous Region, Zhejiang and the northern regions of Jiangsu Province in China.</p><p><b>RESULTS</b>The levels of IFNGR alpha in CHC patients were significantly lower than that of normal controls (NC), especially among patients during the stable stage (P < 0.001), whereas there were no significant differences between CHC in active and stable stages. Among the patients of the three regions, there were no significant differences between patients from Xinjiang and Zhejiang provinces, but both had statistically significant difference compared with the patients from Jiangsu Province (P < 0.001). Plasma IFN-gamma and IL-10 concentrations in CHC patients decreased significantly, IFN-gamma in particular, but there were no significant differences in these levels between various stages of the disease. The IFN-gamma/IL-10 (Th1/Th2) ratio in patients was reversed.</p><p><b>CONCLUSION</b>There may be defects in the IFN-gamma system in chronic HCV infected subjects and a low immune response, which may play an important role in the persistence of HCV infection.</p>

Involvement of TRAIL up-regulation of CD4+, CD8+ T cells in liver injury in chronic hepatitis B / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To describe a novel mechanism for TRAIL up-regulation of CD4+, CD8+ T cells to participate in the pathophysiological process in patients with chronic hepatitis B (CHB).</p><p><b>METHODS</b>The serum levels of soluble TRAIL (sTRAIL), IFN-gamma and membrane bound TRAIL expression on peripheral leucocytes from 58 CHB patients were examined by ELISA and flow cytometry respectively. The levels of TRAIL were compared with the baseline levels of 15 healthy controls, and correlation analysis were performed between ALT, TBil and PT, morphological change in hepatic tissues.</p><p><b>RESULTS</b>The results showed that TRAIL levels on membranes of CD4+, CD8+ T cells in CHB patients were much higher than the healthy controls (P < 0.001), which of CD4+ T cells positively correlated with serum TBil (r=0.354, P = 0.008), Serum IFN-gamma level (r=0.302, P = 0.011) and which of CD8+ T cells positively correlated with serum TBil (r=0.522, P = 0.000), ALT (r=0.393, P = 0.003), PT (r=0.385, P = 0.004), serum IFN-gamma level (r=0.307, P = 0.009). The serum levels of soluble TRAIL only correlated with serum HBeAg expression (r=0.695, P = 0.001).</p><p><b>CONCLUSION</b>These findings suggest that the expression of TRAIL on the membranes of lymphocytes was up-regulated, which may take part in the immunopathogenesis in CHB patients. TRAIL expression can be induced either by virus-specific protein expression or by inflammation cytokine IFN-gamma</p>

Influence of combined cyclosporine A and tacrolimus with 5-fluorouracil on hepatocellular carcinoma rats / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate the effect of combined CsA and FK506 with 5-FU on hepatocellular carcinoma rats.</p><p><b>METHODS</b>A syngeneic rat model of hepatocellular carcinoma was used. Control group (A) underwent 4 ml 5% GS. Treatment group was divided into 3 groups namely, group B: only 5-FU and 5% GS; group C: 5-FU, CsA and 5% GS; group D: 5-FU, FK506 and 5%GS. Cell cycle, apoptosis, necrosis and mitochondrial transmembrane potential were measured by flow cytometry, laser scanning confocal microscopy, and electron transmission microscopy. Statistical analysis was performed by SPSS 10.0 for Windows software. Statistical comparisons were made with ANOVA followed by Dunnett's T3 or LSD test.</p><p><b>RESULTS</b>Compared to the control group, the percentage of apoptotic cells including trifle necrotic cells was significantly higher, and among the treatment group, group D was the highest, and group C was higher than group B. In the treatment group, cell cycle of hepatoma cells was mainly arrested at S phase, but in group D, G0/G1 phase cells were significantly decreased and S phase cells significantly increased. Compared to the control group, mitochondrial transmembrane potential was significantly decreased in the treatment group, among with, group B was the lowest, group C was higher than group D. Morphological changes demonstrated by electron microscopy included dispersed nuclear chromatin, loss of nucleoli, membrane bleeding, cell shrinkage, typical apoptotic bodies and marked swelling of mitochondria in the treatment group. In the control group, however, they were characterized by normal cell ultrastructure.</p><p><b>CONCLUSIONS</b>The present study reveals that 5-FU combined with CsA or FK506 demonstrated a synergistic effect on hepatocellular carcinoma rats. For FK506, the powerful mutual effect is related to the increase of tumor cell's quantity in S phase. Both CsA and FK506 can provide protection on mitochondrial transmembrane potential reduction against hepatoma cells damage from 5-FU.</p>

Modulation and implications in pathogenesis of interferon gamma receptor 1 in patients with chronic hepatitis B / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To study the possible differences in the interferon gamma receptor (IFN-gamma R1) response among a variety of clinical types in patients with chronic hepatitis B (CHB) and implications in pathogenesis.</p><p><b>METHODS</b>The serum level of IFN-gamma and the expression level of IFN-gamma R1 in peripheral leucocytes, from 53 CHB patients, were examined by ELISA and flow cytometry respectively, which were compared with the baseline levels of 15 healthy controls and were performed correlation analysis with alanine aminotransferase (ALT), total bilirubin (TBil) in serum and morphological change in hepatic tissues.</p><p><b>RESULTS</b>The results showed that the level of IFN-gamma R1 (28.89% 11.77%) expressed on the membranes of lymphocytes in CHB patients, which correlated with liver inflammation (r=0.621, P<0.01) and serum TBil level (r=0.575, P<0.01), was much higher than that (9.23% 1.30%) of the healthy controls (Z=3.988, P<0.05), and no obvious difference on the membranes of leucocytes. The serum levels of IFN-gamma in patients with cirrhosis and severe hepatitis were higher than those of healthy controls. And the two was no difference from each other, but the standard deviation in each group was relatively large.</p><p><b>CONCLUSION</b>These findings suggest that the IFN-gamma signal transduction pathway is modulated through up-regulating the expression of IFN-gamma R1 on the membranes of lymphocytes, which takes part in the immuno-pathogenesis in CHB patients.</p>