Background Hypertension is a multigenetic inheritable disease.Gene therapy by regulating gene expression showed long term effects and less side effects,and has been emerged to be a potential and prospective treatment.Objective To investigate the effects of retroviraladeno-associated virus vector containing shRNA targeted on the AT1R gene (rAAV-AT1R-shRNA) on blood preesure(BP) in renal hypertensive rats,and the inhibitory effect of RNA interference (RNAi) on AT1R mRNA expression in renal hypertensive rats.Methods Two-kideny one-clip (2K1C) renal hypertension (RH) were established in SD rats and randomly to receive rAAV-AT1R-shRNA,1.5?109 particles/mL(n=12,ip),as treated group or retroviral vector (rAAV-EGFP),2.9?109 particles/mL,ip,as vehicle group,normal SD rats served as controls (n=12).SBP was measured before and after treatment.Animals were euthanized and blood,brain,heart,liver,kidney,aorta and adrenal gland were collected to identify the sites of rAAV-AT1R-shRNA expression by fluorescence microscope.Angiotensin Ⅱwas assessed by radioimmunology.Results 24 hours after single injection of rAAV-AT1R-shRNA,SBP was reduced by (22.3?5.5)mmHg compared to before intervention (P

Objective To study the relationship between coronary artery disease (CHD ) and interleukin-6 (IL-6), TNF-α and IL-10. Methods Patients with CHD were included in CHD group (n=628) and patients without CHD con?firmed by coronary angiography were selected in control group (n=540) . The recorded data included age, body mass index (BMI) and serum levels of glucose (GLU), total cholesterol (TC), triglyceride (TG), high density lipoprotein-cholesterol (HDL-C) and low density lipoprotein-cholesterol (LDL-C). Meanwhile serum levels of IL-6, TNF-α, IL-10 were determined by ELISA. Results Serum levels of BMI, LDL-C, TG were higher in CHD group than those in control group(P<0.01). The serum level of HDL-C was significantly lower in CHD group than that in control group(P<0.01). Serum levels of IL-6(P<0.05)and TNF-α(P<0.01)were higher in CHD group than those in control group. The serum level of IL-10 was significant?ly lower in CHD group than that in control group(P<0.01). Conclusion IL-6 and TNF-αare involved in the develop?ment of CHD. IL-10 can inhibit inflammation and protect vessel integrity.

Objective To study the role of the oxidative stress in the development of wound healing and observe the effect of the antioxidant PTD-SOD on damage and inflammation reaction after mechanical wound. Methods In this experiment,acute wound healing model by removal the whole layer dorsal skin of the mice was prepared,SOD(3 000 U and 6 000 U)and the fusion protein PTD-SOD with different concentrations(1 000 U,3 000 U,6 000 U and 10 000 U)were used to deal with the wounds continuously for 13 days.The mice were divided into different concentration SOD treatment group and PTD-SOD treatment group,model control group,physiological saline treatment group and compound iodine solution control group.The wound healing situation and healing percentage of the fight and left skin wounds of each mouse in every group was recorded every day.At day 14 after wound,the wound healing skin of each group was removed and some were used to make 10%tissues homogenate for detecting the activities of superoxide dismutase(SOD),catalase(CAT),glutathione peroxidase(GSH-Px)and contents of malondialdehyde(MDA)and hydroxyproline(Hyp);in the meantime,the other removed skin were fixed in 10% formalin for observing the histopathological changes of the tissues. Results Compared with the model control group,the physiological saline treatment group and the compound iodine solution control group,the skin wound healing percentage was significantly(P<0.05 or P<0.01)improved,with increase of the activities of SOD,CAT,GSH-Px and contents of Hyp (P<0.05 or P<0.01)and decrease of MDA(P<0.05 or P<0.01) in the SOD groups or PTD-SOD groups (except for 10 000 U PTD-SOD group).When compared with the physiological saline treatment group or the compound iodine solution treatment group,the effect was similar to the model control group.In comparison to the SOD groups,under the same dosage and environment condition,the PTD-SOD groups were much better than SOD groups with regard to promoting skin wound healing percentage,increasing activities of antioxidases and contents of Hyp,decreasing contents of MDA.Among the PTD-SOD groups,the effect of high dosage 10 000 U on promoting skin wound healing was declined. Conclusions The oxidative stress may playan important role in the development of wound healing.Proper application of treatment with antioxidants is a alternative strategy in the early stage of wound.PTD-SOD is able to prevent the oxidative stress damage,inhibit inflammatory infiltration and promote skin wound healing efficiently.

Objective To explore the relationship between B-type natriuretic peptide (BNP) and fibrinogen (Fib) in the elderly patients with coronary heart disease (CHD). Methods According to the level of BNP,325 subjects aged more than 65 years were divided into four groups,BNP:≤32.4ng/Lgroup,32.5～62.3 ng/Lgroup,62.4～162.5 ng/Lgroup,＞162.5 ng/L group.The levels of left ventricular ejection fraction (LVEF),left ventricular end-diastole dimension (LVEDD) and Fib were compared among the groups. Results With increasing BNP level,the LVEF was gradually decreased [(63.3±8.2)％,(59.6±7.4)％,(57.9±9.7)％,(55.2±10.6)％] and the level of LVEDD was increased[(50.5±4.7)mm,(51.4±5.4)mm,(52.8±5.7)mm,(54.2±5.9)mm ;F=11.54,6.76,both P＜0.01],the level of Fib was enhanced [(3.4±0.7)g/L,(3.7±0.8)g/L,(3.8±1.1)g/L,(4.2±1.0)g/L; F=8.02,P＜0.01].The level of BNP had positive correlation with Fib (r=0.25,P＜0.01). Multi-linear regression analysis showed that Fib and LVEF were influencing factors of BNP (t =2.92, - 3.15, both P ＜ 0.01 ). Conclusions The BNP level is helpful to diagnose heart failure and assess heart function.There is an association between BNP and Fib level.

Objective To explore the relationship between blood lipids and creatinine clearance (Ccr)in aged patients with coronary heart disease(CHD).Methods According to coronary artery angiography(CAG)and Ccr,784 subjects aged 65 years and over were divided into groups and Their lipid levels were compared.Results With increasing lesion of coronary branches,the level of Ccr was obviously decreased(F=5.35,P＜0.01).The level of apolipoprotein A1(ApoA1)was reduced and apolipoprotein B(ApoB)enhanced in renal function of moderate-severe injury(CCr＜0.83 ml ·s-1 · 1.73m-2)(F=5.31,F=4.91,both P＜0.01).The level of ApoA1 was decreased(F=3.52,P＜0.05)and Apo B increased(F=5.65,P＜0.01)in male renal function of moderate-severe injury.The levels of ApoA1 decreased(F=5.79,P＜0.01),ApoB increased(F=4.56,P＜0.05)and high density lipoprotein-cholesterol reduced(F=3.39,P＜0.05)in CHD with renal group of moderatesevere injury.Multi-logistic regression analysis showed that Ccr,HDL-C,ApoA1 and ApoB were influencing factors of CHD.Conclusions Abnormal blood lipid and renal dysfunction are risk factors of CHD,dyslipidemia may induce renal dysfunction.It is important to control lipids and improve other organ functions in the aged patients with CHD.

Objective To explore the correlation of B-type natriuretic peptide (BNP) with blood lipids and renal function in elderly patients with coronary heart disease (CHD).Methods According to the four quantiles of BNP level,325 subjects aged 65 years and over were divided into four groups:group A (≤32.4 ng/L),group B (32.5 ng/L),group C (62.4 ng/L),group D (＞ 162.5 ng/L).The left ventricular ejection fraction (LVEF),left ventricular end-diastole dimension (LVEDD),levels of blood lipids and renal function were compared among the groups.Results With the increase of BNP level,the LVEF was gradually decreased [(63.3±8.2) ％,(59.6±7.4) ％,(57.9±9.7)％,(55.2±10.6)％,respectively,F=11.54,P＜0.01] and the LVEDD was gradually increased (F=6.76,P＜0.01),the level of triglyceride (TG) was gradually decreased (F=2.73,P ＜0.05) in group A,B,C and D.Creatinine clearance was gradually decreased [(1.24±0.31) ml ·s-1 · 1.73 m-2,(1.21±0.31) ml· s-1 · 1.73 m-2,(1.24±0.29) ml · s-1 · 1.73 m 2,(1.09± 0.33) ml · s-1 · 1.73 m-2,respectively,F=3.62,P＜0.05],and blood urea nitrogen was gradually increased (F=4.43,P＜0.05) in group A,B,C and D.Multi-linear regression analysis showed that levels of total cholesterol,triglyceride,low density lipoprotein-cholesterol,high density lipoproteincholesterol,very low density lipoprotein-cholesterol,creatinine clearance and blood urea nitrogen were influencing factors for BNP (all P＜0.05).Conclusions BNP level can be used as a sensitive indicator for the diagnosis of heart failure and the assessment of its severity.The levels of blood lipids and renal function are associated with BNP.

Objective To investigate the relationship between matrix metalloproteinase-3 (MMP-3) gene promoter polymorphisms 5A/6A and the restenosis after percutaneous coronary intervention (PCI). Methods A total of 437 patients with PCI were selected in this study. Patients were divided into mutant genotype group (5A/5A+5A/6A, n=136) and wild genotype group (6A/6A, n=301) according to MMP-3 polymorphism. The angiography and clinic data were collected before and after coronary angiography in two groups of patients. The serum level MMP-3 and genotype analysis were compared be-tween two groups. Results There was no significant difference in the restenosis rate between two groups (42.2%vs 33.1%, P>0.05). The restenosis degree was significantly higher in wild genotype group than that in mutant genotype group (56.28%± 11.10%vs 36.00%±10.17%, P＜0.01). There was no significant difference in the serum level of MMP-3 between two groups (13.38μg/L ± 3.00μg/L vs 12.33μg/L ± 2.96μg/L, P>0.05). There was a higher restenosis rate in patients carrying 6A al-lele than that of patients carrying 5A allele (P＜0.05). Carrying wild genotypes are risk factors for restenosis after PCI. Con-clusion Patients carrying 6A allele have significantly higher risk of resteonsis than patients carrying 5A allele.

Aminopeptidase H (APH) is an universally distributed aminoendopeptidase in the tissue of many organism. However, it is hard to investigate its mechanism underlying the catalysis and the function in cell. In this paper, the full DNA sequence of this enzyme was cloned from chicken liver, then subcloned to the vector pET22 b(+). The recombined vector was transformed into E. coli Rosetta(DE3), and the APH gene was expressed by the induction of IPTG. It was found the recombinant protein exhibited same mo lecular weight as authentic APH on SDS-PAGE analysis; the expression level increased with induction time and approached maximum of 94.7 mg/L till 6 hours, which contained 16.7% of the total protein. Moreover, this recombinant protein showed similar prop erties of subunit composition, thermal stability and optimum pH with native APH, based on the enzymatic assay, purification and analysis of enzymological properties. Therefore, it is confirmed that APH was expressed in this prokaryote system with a high-level of 1636 u/L aminopeptidase activity. These results would help to elucidate the catalysis mechanism and biological function of APH by providing enough material.
Aminopeptidases ; biosynthesis ; genetics ; Animals ; Chickens ; Endopeptidases ; biosynthesis ; genetics ; Enzyme Stability ; Escherichia coli ; genetics ; metabolism ; Recombinant Proteins ; biosynthesis ; genetics ; isolation & purification

OBJECTIVE: To assess the association of 5A/6A polymorphism in the promoter region of MMP3 gene with the stability of extracellular matrix of atherosclerotic plaque. METHODS: Clinical data of 776 consecutive patients undergoing percutaneous coronary intervention (PCI) was reviewed. MMP3 gene polymorphisms and serum level of MMP3 for the second admission were collected. The target gene fragment containing MMP3 promoter region was transfected into HepG2 vector cells. The influence of the polymorphism on the expression of the MMP3 gene was determined in vitro. RESULTS: Compared with the first admission data, the proportion of mutant MMP3 genotypes (5A/5A+5A/6A) was significantly higher in patients with acute myocardial infarction (AMI) compared with the control group (37.6% vs. 24.9%, P<0.01). 64.1% of the patients carrying the 5A allele had AMI, whereas only 50.11% of those carrying the 6A allele had AMI (P<0.01). The proportion of wild-type MMP3 genotype (6A/6A) was significantly higher in the stenotic group compared with the non-restenosis group (79.5% vs. 66.5%, P<0.01). Restenosis has occurred in 9.5% of patients harboring the 5A allele compared with 16.2% in those carrying the 6A allele (P<0.01). In addition, serum level of MMP3 in the restenosis group was significantly lower than that of the non-restenosis group (P<0.01). In vitro studies confirmed that the expression of pGL2-Basic/6A was significantly lower than that of pGL2-Basic/5A. CONCLUSION The 5A/6A polymorphism in the promoter region of the MMP3 gene may influence its transcriptional activity and impact on the degradation or push-up of extracellular matrix, resulting in a difference in the stability of atherosclerosis plaques, which in turn may induce different pathological processes in AMI or restenosis after stenting.
Case-Control Studies ; Extracellular Matrix ; Genetic Predisposition to Disease ; Genotype ; Humans ; Matrix Metalloproteinase 3 ; genetics ; Percutaneous Coronary Intervention ; Plaque, Atherosclerotic ; genetics ; Polymorphism, Genetic ; Promoter Regions, Genetic

Objective To investigate the clinical features of patients with heart failure and the safety and efficacy of noninvasive ventilator in patients with heart failure.Methods Sequentially enrolled 65 patients who were diagnosed with decompensated heart failure in Tianjin Chest Hospital Heart Center from October 2016 to October 2017 and who had acute heart failure during hospitalization requiring noninvasive ventilator,were divided into the HF-PEF group (n=19) and HF-REF group (n=46).The clinical data of the two groups and the observation indexes before and after the application of the non-invasive ventilator were compared.Results Comparing the admission data of the two groups,the proportion of patients with hypertension (57.9％ vs 21.7％,P=0.005) and LVEF(％) (53.00±4.85 vs 33.07±7.24,P＜0.01)were significantly higher in the HF-PEF group than those in the HF-REF group;LVEDD (mm) in the HFPEF group was significantly lower than that in the HF-REF group (50.00±5.23 vs 63.82±8.95,P＜0.01).In the two groups of patients with acute left heart failure,blood lactate levels (mmol/L) in the HF-PEF group (4.20±1.06 vs 2.02±0.88,P＜0.05) and systolic blood pressure (mmHg) (151.32±43.40 vs 117.90± 19.55,P＜0.05) were significantly higher than those in the HF-REF group.After the application of non-invasive ventilator,systolic blood pressure (mmHg) (34.38±9.36 vs 16.94±5.19,P=0.038) and PaCO2 (mmHg)(2.49±0.98 vs-0.06±0.00,P=0.025),and lactic acid (mmol/L) (2.06±0.67 vs 0.04±0.01,P=0.001) were significantly lower in the HF-PEF group than those in the HF-REF group.While the NT-proBNP level (ng/L) (13 064.90±1 963.83 vs 11 687.13±1 028.03,P=0.848) did not decrease significantly,and the time of non-invasive ventilator application (h)was significantly longer than that in the HF-REF group (152.74±10.61 vs 71.03±10.41,P=0.013).Conclusions Hypertension is the main cause of HF-PEF group.The systolic blood pressure and blood lactate level in HF-PEF patients with acute left heart failure are significantly higher than HF-REF patients.Non-invasive ventilator is also safe and effective for the treatment of acute left heart failure in HF-PEF patients,but HF-PEF patients with acute left heart failure have a longer clinical remission time.