Objective To investigate the influence of MDR polymorphism on CsA-associated hepatotoxicity. Methods PCR/RFLP was used to analyze the genotypes of MDR exon26 in 187 recipients. CsA-associated hepatotoxicity was evaluated among groups being classified according to the genotypes. Results MDR exon26 takes 3 genotypes:CC,CT and TT, in the following ratio: 29.4%(55/187), 40.1%(75/187), 30. 5%(57/187). Among the same range of whole blood CsA concentration, incidence of CsA-associated hepatotoxicity is markedly higher in the CT/TT group than that in in patients with CsA-associated hepatotoxicity between the CC group and the CT/TT group is signifiimportant role in the development of CsA-associated hepatotoxicity.

Objective To study expression of matrix metalloproteinase‐9(MMP‐9) protein and its mRNA in breast cancer and e‐valuate its significance in the occurrence ,development and metastasis of breast cancer .Methods The protein expression of MMP‐9 breast cancer were detected by using immunohistochemistry and the expression of mRNA were detected by reverse transcription polymerase chain reaction(RT‐PCR)Results The positive rate of MMP‐9 protein expression in 56 cases of breast cancer was 69 .6% (39/56) ,while in benign breast diseases was 20% (6/30) ,which were significantly different(P<0 .05) .The levels of MMP‐9 mRNA were significantly higher in the patients with breast cancer than those in benign breast diseases(P<0 .05) ,which were 0 .914 2 ± 0 .108 1 and 0 .379 4 ± 0 .0428 respectively .Conclusion The MMP‐9 protein and mRNA expression in human breast cancers are positively correlated with the stage and lymph node metastasis .Expression of MMP‐9 could be used as an indicator for the ability of invasion and metastasis of breast cancer .

Objective To investigate the levels of platelet-derived growth factor-BB (PDGF-BB) and its role in the pathogenesis of the patients with hepatic diseases.Methods The levels of serum PDGF-BB in 30 chronic patients (hepatitis B,26; hepatitis C,4),24 patients with post-hepatitis cirrhosis,30 patients with primary hepatocellular carcinoma,and 20 normal controls were measured by ELISA.The levels of PDGF-B mRNA in peripheral blood mononuclear cells were measured by reverse transcription polymerase chain reaction (RT-PCR).Results The serum levels of PDGF-BB and the levels of PDGF-B mRNA in peripheral blood mononuclear cells were significantly higher in the patients with hepatic diseases (F=1774.40,1037.42,P

Objective To establish the chip technology‐based real‐time PCR (RT‐PCR) platform and to apply it in the viral loads detection of HIV‐1 and HCV .Methods Based on the primers designed to aim at the conversed regions of HIV‐1 and HCV , The gene chip tube was prepared ,and the RT‐PCR reaction system was established for the simultaneous determination of viral loads .And the melting curves were used to distinguish viral species .The sensitivity and specificity of the method were estimated , and performance of the method was verified by using clinical samples .Results The specificity of the method was good .The lowest detectable limit of the detection method of HCV and HIV‐1 was 1 × 103 copy/mL .The clinical samples with viral loads around 1 × 103 -1 × 106 copy/mL could be detected accurately .Conclusion The method provides a new idea for the detection of HCV and HIV‐1 .

Previous work from this laboratory has cloned a novel gene NOR1 and showed its extensive expression in normal tissues and down-regulation in carcinomas. To further investigate its downstream target genes and better understand its function, NOR1 was over-expressed in HepG2 hepatoma cells and global changes in gene expressions from a stable line were identified by cDNA microarrays. The results discovered 59 genes up-regulated in these cells compared with the original cells, including Grb2, HBP17,TNFRSF11B genes that have been implicated in tumorigenesis and cancer development. In addition, 103 down-regalated genes were also identified, including genes encoding Bik, MAP2K6 and ZFP95 proteins. The expression patterns of certain genes identified by microarrays were validated by quantitative real-time PCR and the results showed that expression difference were statistically significant (P< 0.05). These data suggest that NOR1 may influence the biology and cancerous behaviors of HepG2 cells by regulating expression of a set of genes involved in signal traasduction, cell cycle regulation, transcription and Wanslation controls.

Objective To construct the eukaryotic expression vector of pcDNA3.1(+)/NOR1 and analyze the effect of NOR1 on the liver cancer cells. Methods NOR1cDNA was cloned into eukaryotic expression vector pcDNA3.1(+), recombinant eukaryotic expressing plasmid pcDNA3.1(+)/NOR1 was transiently introduced into human liver cancer cell line HepG2 mediated by cation iron lipofectamin.The biology effect of NOR1 on the liver cancer cells was analyzed through the MTT test, trypan blue exclusion assay and flowcytometric analysis. Results The eukaryotic expression vector of pcDNA3.1(+)/NOR1 was successfully constructed.The liver cancer cell growth rate was obviously slow after it was transfected by recombinant plasmid pcDNA3.1(+)/NOR1 and the cell cycle from G0/G1 to S distinctly prolong.Conclusions Recombinant plasmid pcDNA3.1(+)/NOR1 can express in HepG2 cells and affect the growth of HepG2 cells.

BACKGROUND:In vitro studies have demonstrated that basic fibroblast growth factor (bFGF) promote the differentiation of bone marrow mesenchymal stem cells (BMSCs) into cardiomyocyte-like cells. However, it is unclear whether coronary venous retroperfusion of bFGF stimulates BMSCs differentiation in vivo. OBJECTIVE:To evaluate the effects of coronary venous retroperfusion of bFGF on BMSCs differentiation in vivo. METHODS:BMSCs from 12 dogs were isolated by density gradient centrifugation and expanded in vitro. These cells were transfected by enhanced green fluorescence protein (EGFP) lentiviral vector and the transfection efficiency was analyzed. Acute myocardial infarction was induced by ligation of left anterior descending coronary artery. After 1 week, 10 survival animals were randomized to BMSCs group (n=5) and bFGF+BMSCs group (n=5). bFGF-and EGFP-positive BMSCs were reversely infused via coronary vein using over-the-wire bal oon catheter. One week after infusion, the number of EGFP-positive cells co-staining factor VIII and troponin I was compared between the two groups by immunofluorescence method. RESULTS AND CONCLUSION:BMSCs were successful y transfected by EGFP and the transfection efficiency was 85%. Immunofluorescence showed that EGFP-positive BMSCs were observed in 23.5%of slides. There were more EGFP-positive cells co-staining VIII and troponin I in the bFGF+BMSCs group than in the BMSCs group (P<0.05). Thus, the coronary venous retroperfusion of bFGF enhances the differentiation of BMSCs into vascular endothelial cells and cardiomyocytes. Combined delivery of bFGF and BMSCs can exert a synergistic effect to promote cardiac repair.

OBJECTIVE: To investigate the mechanisms by which MecA gene expression leads to β-lactam resistance in methicillin-resistant Staphylococcus aureus (MRSA), and to study the resistance mechanism of MRSA at the molecular level. METHODS: A variety of molecular biological techniques were employed, including screening MRSA using cefoxitin paper disk method, extraction of MRSA mRNA, reverse transcription into cDNA, real-time fluorescence PCR for quantitation of MecA gene expression, and agar dilution method for assessment of minimum inhibitory concentrations in MRSA treated with cefoxitin, oxacillin, vancomycin, or linezolid. RESULTS: According to the level of resistance of MRSA to cefoxitin, 40 MRSA strains were divided into a low resistance group (n=12), a middle resistance group (n=15), and a high resistance group (n=13). The expression level of the MecA gene in the low resistance group, the middle resistance group, and the high resistance group was 58.87±30.30, 363.37±200.05, and 1257.72±446.63, respectively. MRSA resistance to cefoxitin and oxacillin was 100%; MRSA resistance to vancomycin or linezolid could not be detected. For all 40 MRSA strains the MIC90 for vancomycin was 2.0 μg/mL. CONCLUSION MecA gene expression levels may correlate with the MRSA level of resistance to cefoxitin within a certain range of concentration.
Anti-Bacterial Agents ; pharmacology ; Bacterial Proteins ; genetics ; metabolism ; Cefoxitin ; pharmacology ; Drug Resistance, Multiple, Bacterial ; Methicillin-Resistant Staphylococcus aureus ; drug effects ; genetics ; metabolism ; Microbial Sensitivity Tests ; methods ; Oxacillin ; pharmacology ; Penicillin-Binding Proteins

10.3969/j.issn.2095-4344.2013.24.015

Objective To understand the application of drug eluting stent (DES) in renal functional insufficient patients, and to assess its safety and effectiveness, especially the occurrence of stent thrombosis(ST) after DES implantation and its related factors. Methods The subjects were all the patients underwent percutaneous coronary intervention (PCI) as well as at least one DES admitted to Beijing Anzhen Hospital consecutively from July 2003 to June 2005. All patients were divided into 2 groups: Group Ⅰ with normal or mild renal insufficiency (Ccr≥60 ml/min),and Group Ⅱ with moderate to severe renal functional insufficiency (Ccr ＜ 60 ml/min). All of the clinical, angiography and intervention data were recorded. ST was adjudicated by the definition of ARC Dublin. The rates of MACCE in hospital and during the follow-up between the 2 groups were compared. Results There were 2377 patients enrolled in the study, of which 2020 ( 85.0% ) patients presented Ccr ≥ 60 ml/min, and 357( 15.0% ) presented Ccr ＜ 60ml/min. The case fatality during follow-up in group Ⅰ was significantly higher than that in group Ⅱ (4. 5% vs. 1.2%, P ＜ 0. 001 ). However, the incidences of ST were not significantly different between each stage of disease( P ＞0. 05 ). The results from Cox regression showed that renal functional insufficiency was not a risk factor of death,whereas multivessel coronary artery disease [OR = 1. 929(95% CI: 1. 178 -3. 157),P =0. 009] ,diabetes [OR = 1. 914(95% CI:1. 055 -3. 470) ,P =0. 033] and age [OR = 1. 051 (95% CI:1. 005 -1. 099 ) ,P = 0. 030] were independent risk factor of death after DES implantation in patients with moderate to severe renal functional insufficiency. Conclusions Compared with normal renal function or mild renal patients, the longterm case fatality is higher in moderate and severe renal functional insufficiency patients. However, the higher case fatality does not due to the increase of ST.