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 .

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 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 .

10.3969/j.issn.2095-4344.2013.24.015

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

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 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.

Objective To evaluate the short-term and long-term outcomes of patients with ST-segment elevation myocardial infarction (STEMI) compared with those with non-STEMI after percutaneous coronary intervention (PCI). Method The DESIRE Ⅱ (Drug-Eluting Stent Impact on Revascularization Ⅱ) was a single-center registered retrospective study of coronary revascularization in our institution between July 2003 and September 2009.Data of demographics, clinical features and revascularization record of STEMI and non-STEMI patients from the DESIRE Ⅱ trial were analyzed. The patients were followed up in OPD or by telephone after discharge. MACCE (major adverse cardiocerebral events) including death, neo-myocardial infarction, stroke and revascularization were recorded. The clinical outcomes of patients of two types were evaluated. Results There were 6005 patients studied with a median follow-up of 566 days. A total of 1009 STEMI and non-STEMI patients were analyzed. The patients with non-STEMI ( n = 206) had higher prevalence of hypertension and history of higher frequency of myocardial infarction as well as revascularization compared with patients with STEMI ( n = 803). The patients with non-STEMI had higher ratio of treatment for multivessel disease (43.7% vs. 34.4%, P = 0.039). There were no significant differences in in-hospital mortality and long-term outcomes (one year survival rate: 96% vs. 98%)between patients with STEMI and non-STEMI. The predictors of 1-year mortality were LVEF and blood creatine.Conclusions Despite different chnical features, patients with STEMI and non-STEMI after PCI had similar both short-term and long-term outcomes.

Objective To assess the prevalence, and characteristics, and in-hospital and long-term prognosis of coronary artery disease (CAD) with metabolic syndrome, and to determine the factors affecting the prognosis of CAD most. Method The DESIRE (drug-eluting stent impact on revascularization) registry covered a database of 2368 patients with coronary artery disease (CAD) treated with percutaneous coronary intervention (PCI) or coronary artery bypass graft (CABG) in a period between July 2003 and September 2004. The median long-term follow-up time was 3.5 years ranged from 293 to 1855 days. The metabolic syndrome (MS) was diagnosed on the Definition of the Metabolic Syndrome modified by the Adult Treatment Panel (ATP) Ⅲ in 2005, by using the body mass index (BMI) instead of waist circumference. The relationship between metabolic syndrome (MS) and the incidences of major adverse cardiac as well as cerebral events (MACCE) in a large cohort of patients treated for revascularization was analyzed by using logistic analysis and Cox regression with SPSS 11.0 software. Results The Ms was present in 45.6% patients (high fast glucose (FG) in 44.5% patients, high triglycerides (TG) in 45.0% patients, low high density lipoprotein (HDL) in 50.8% patients, high BP in 61.4% patients, high BMI in60.7% patients). After follow-up, the ratio of MACCE in CAD patients with metabolic syndrome increased significantly (18.9% vs. 15.6%, P <0.036). The most dangerous factors of MS were high FG, hypertension and obesity (OR=1.787, 95%CI=1.132-2.845, P =0.014). Conclusions The MS contributes the high risk factors of MACCE in CAD patients with or without diabetes. The most dangerous combination of risk factors in MS is the combination of high FG, hypertension and obesity.

Objective To evaluate the effect of clopidogrel premedication on in-hospital major adverse cardiovascular and cerebral events (MACCE) and bleeding outcomes before coronary artery bypass graft surgery (CABG). Method A total of 2021 patients who underwent CABG from July 2003 to September 2005 were divided into either clopidogrel ( n = 479) or no clopidogrel (1542) group before CABG. Patients with clopidogrel administration ( n = 479) were subdivided into < 5 d ( n = 154) ,5～7d(n = 183)and >7d(n = 142) group according to timing of clopidogrel withdrawal before surgery. In-hospital MACCE and perioperative bleeding outcomes were analyzed among groups. Results Patients who took clopidogrel before surgery had nonsignificantly rates of bleeding and in-hospital MACCE compared with those patients not administered clopidogrel. No differences were found about the incidence of total bleeding,minor bleeding,transfusions of red blood cells,fresh frozen plasma,whole blood and in-hosptial MACCE among three subgroups.The < 5 d group had higher incidence of major bleeding and more platelets transfusions than 5 ～ 7 d [47.8% vs. 31.9%,P < 0.017; (0.08 ±0.38) U vs. (0.00±0.00) U,P <0.017,respectively]and >7 d group [47.8% vs. 20.3%,P <0.017; (0.08±0.38) U vs. (0.00±0.00) U,P <0.017,respectively). However,there were no significant differences between 5 ～ 7 d and > 7 d group ( P > 0.05). Conclusions Gopidogrel administration before CABG does not increase the incidence of in-hospital MACCE events. However,the perioperative risk of bleeding will rise if the patients withhold clopidogrel less than five days before surgery.