To prepare HIV-1 Vif and hAPOBEC3G and to produce their antibodies, the full length gene fragment of HIV-1 Vif was amplified by PCR from a plasmid of HIV-1 NL4.3 cDNA, and the APOBEC3G gene was obtained by RT-PCR from the total RNA of H9 cells. The resulting DNA construct was cloned into a prokaryotic expression vector (pET-32a). Recombinant pET-vif and pET-APOBEC3G were expressed respectively in Eserichia coli BL21 (DE3) as an insoluble protein. The vector also contained a six-histidine tag at the C-terminus for convenient purification and detection. To express and purify the HIV-1 Vif and hAPOBEC3G in E. coli cells, the accuracy of inserted gene and specificity of proteins were detected by the two enzyme digestion method, SDS-PAGE, and Western blotting. Rabbits were then immunized by Vif or APOBEC3G protein and serum samples were tested by indirect ELISA to determine the level of antibodies. Immunoenzyme and immunofluorescence assays were performed to identify the specificity of polyclonal antibodies. The titer of the anti-Vif antibodies was 1:204800, and that of the anti-APOBEC3G antibodies was 1:102400. Thus the antibodies could detect the antigen expression in the cells, demonstrating that fusion proteins with high purity and their corresponding polyclonal antibodies with high titer and specificity were achieved.

Objective To discuss the effects of blood purification on the patients with multiple organ dysfunction syndrome. Methods Technique of blood purification was applied to the 22 patients with multiple organ dysfunction syndrome. The patients were presented to the Fushun Central Hospital ,between 2003 to 2006. Liver function,kidney function before treatment and after treatment were observed. Results After ttreatment with blood purification,The liver function,kidney function were ameliorated (P

Animals ; Antigens, CD ; chemistry ; genetics ; metabolism ; Antiviral Agents ; chemistry ; metabolism ; GPI-Linked Proteins ; HIV Infections ; metabolism ; virology ; HIV-1 ; genetics ; physiology ; Human Immunodeficiency Virus Proteins ; genetics ; metabolism ; Humans ; Membrane Glycoproteins ; chemistry ; genetics ; metabolism ; Protein Binding ; Viral Regulatory and Accessory Proteins ; genetics ; metabolism ; Virus Shedding

Objective To observe the anti-influenza effect of a Chinese medicinal herb-Antiviral Agent No.1. Methods To study t he anti-viral effect of Antiviral Agent No.1 by means of the technique of cel l culture and using Ribavirin as a positive control. Results In MDC K cu lture, Antiviral Agent No.1 was found to be a potential inhibitor of influenza A 3 virus in a concentration-dependent manner, with a TC50 of 60.53 mg*m l-1. When drug was added 2 hours post virus infection, the EC50(TI) was 5.14 mg*ml-1(11.78); while drug was added 2 hours before infection, t he EC50(TI) was 5.20 mg*ml-1(11.65). Conclusions Antiv iral Agent No.1 had a significant anti-influenza effect on type A3 in MDCK.

Apolipoprotein B mRNA-editing enzyme catalytic polypeptide-like 3G (APOBEC3G) cDNA was amplified from total RNA prepared from nonpermissive H9 cells by RT-PCR. APOBEC3G cDNA is 1155nt long, encoding 384 amino acids. The APOBEC3G gene was then cloned into the eukaryotic expression vector pEGFP-C3. The generated pEGFP-3G construct was then transfected into CD4+ HeLa cell to determine the expression and the subcellular localization of GFP-APOBEC3G fusion protein. Under CLSM the localization of the expressed GFP-APOBEC3G in the cytoplasm of CD4+ HeLa cells was observed.
APOBEC-3G Deaminase ; Cell Line ; Cytidine Deaminase ; Cytoplasm ; metabolism ; DNA, Complementary ; genetics ; isolation & purification ; metabolism ; Green Fluorescent Proteins ; genetics ; metabolism ; HeLa Cells ; Humans ; Microscopy, Confocal ; methods ; Nucleoside Deaminases ; genetics ; metabolism ; Recombinant Fusion Proteins ; genetics ; metabolism ; Repressor Proteins ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

Objective To evaluate the beneficial effects of early coronary reperfusion on left ventricular remodeling (LVRM) and systolic function in patients with acute myocardial infarction (AMI).Methods Eighty-one patients with first AMI in the convalescent stage and having undergone left ventriculography (LVG) and coronary arteriography (CAG) were divided into four groups: the anterolateral wall (ALW) myocardial infarction (MI) non-reperfusion (n=20) and reperfusion (n=21), and inferoposterial wall (IPW) MI non-reperfusion (n=20) and reperfusion (n=20), according to infarct location and early treatment with or without successful coronary reperfusion therapy within 6 hours after onset of symptoms. By LVG, the parameters of LVRM and systolic function in the four MI groups were analyzed and compared with those in normal group (n=25) and between the two reperfusion and non-reperfusion MI groups.Results In both ALW and IPW MI non-reperfusion groups, the left ventricular (LV) end-diastolic volume (EDV), circumference (EDC), short-axis dimension (EDD), short to long axis ratio (ED-D/L), sphericity index (ED-SI) and end-systolic volume (ESV) were all significantly increased (P<0.01-0.001), while LV ejection fractions (LVEF) were significantly decreased (both P<0.001) when compared with those of normal group; and the increase in ESV and decrease in LVEF were both significantly greater in ALW than in IPW MI groups (both P<0.01).In both ALW and IPW MI reperfusion groups, however, the EDV, EDD, ESV, as well as the extent and severity of regional wall motion abnormality (RWMA) were significantly smaller (P<0.05-0.001), while LVEF were significantly higher (P<0.01-0.001) when compared with those in the two non-reperfusion MI groups respectively. There were no longer significant differences in LVEF and ESV between ALW and IPW MI groups (both P>0.05). The EDC in IPW MI reperfusion group and the ED-D/L and ED-SI in ALW MI reperfusion group were also significantly reduced compared with those in the two non-reperfusion MI groups respectively (P<0.05-0.001). All the above parameters in the two reperfusion MI group were decreased to the normal in comparison with normal group except ESV and LVEF, and ED-D/L and ED-SI in IPW MI group.Conclusion It was indicated that in both ALW and IPW MI non-reperfusion groups, LVRM had occurred in convalescent stage of AMI with an increase in EDV and EDC, spherical change in LV shape, and accompanying reduction in LV systolic function; and early coronary reperfusion in AMI could reduce the extent and severity of RWMA, prevent from LV enlargement and remodeling, and preserve or improve LV systolic function with more prominence in ALW MI.

OBJECTIVESTo compare the effects of losartan, enalapril and their combination in the prevention of left ventricular remodeling (LVRM) after acute myocardial infarction (AMI) in the rat.

METHODSAMI model was induced in female SD rats by ligating left coronary artery. Forty-eight hours after the procedure, 83 surviving rats were randomized into one of the following 4 groups : 1) AMI control group (n = 19), 2) losartan group (n = 22, 3 mg x kg(-1) x d(-1)), 3) enalapril group (n = 20, 1 mg x kg(-1) x d(-1)), 4) losartan-enalapril combinative group (n = 22, 3 and 1 mg x kg(-1) x d(-1) respectively). 5) Sham-operated group (n = 10) and 6) normal rats group (n = 10) were selected randomly to serve as non-infarction controls. Losartan and enalapril were delivered by direct gastric gavage. After 4 weeks of medical therapy, hemodynamic studies were performed in each group, then the rat hearts were fixed with 10% formalin and pathologic analysis on them was performed. Complete experimental data was obtained in 56 rats, comprising 1) AMI controls (n = 11), 2) losartan group (n = 10), 3) enalapril group (n = 10), 4) the combination of losartan and enalapril group (n = 11), 5) sham-operated group (n = 6) and 6) normal controls (n = 8).

RESULTSThere were no significant differences among the 4 AMI groups in MI size (41.7% to approximately 43.4%, all P > 0.05). Compared with sham group, the left ventricular (LV) end diastolic pressure (LVEDP), volume (LVV), long and short axis length (L and D), as well as LV absolute and relative weight (LVAW and LVRW) in AMI group were all significantly increased (P < 0.05 to approximately 0.001); whereas the maximum left ventricular pressure rising and dropping rates (+/- dp/dt) and their corrected values by LV systolic pressure (+/- dp/dt/LVSP) were significantly reduced (all P < 0.001), indicating LVRM occurred and LV systolic and diastolic function impaired after AMI. Compared with AMI group, LVEDP, LVV, LVAW and LVRW were all significantly decreased (P < 0.05 to approximately 0.001); while +/- dp/dt/LVSP were significantly enhanced in all 3 treatment groups (P < 0.05 to approximately 0.001) except -dp/dt/LVSP in losartan group (P > 0.05). There were no significant differences in the above indices among the 3 treatment groups (all P > 0.05).

CONCLUSIONBoth losartan and enalapril can prevent from LVRM after AMI in the rat and improve LV function with equivalent effects. There seems no additive effect when the 2 drugs are used in combination.

Animals ; Antihypertensive Agents ; pharmacology ; Drug Synergism ; Enalapril ; pharmacology ; Female ; Losartan ; pharmacology ; Myocardial Infarction ; physiopathology ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Ventricular Function, Left ; drug effects ; Ventricular Remodeling ; drug effects

BACKGROUND: With social development, the average life span is prolonged. The issue of aging has increasingly attracted people's attention. It is necessary to promote the health status and the guality of life of the elderly..OBJECTIVE: To understand the status and distribution features of several common diseases which influence the quality of life of elderly.DESIGN: Random cluster sampling methods and a cross-section survey.SETTING: Beijing Institute of Geriatrics of Beijing Hospital.PARTICIPANTS:Totally 1 558 old people aged ≥60 years living in 10 communities affiliated to Jianguomen district office of the Eastern District of Beijing City,and 8 villages affiliated to Xiangyang and Henan Villagers'Commissions of Shunyi County of Beijing from May 1997 and October 1997 were recruited.In fact,1 434 persons responded,with a response rate being 92％,319 males and 498 females came from the city;242 males and 375 females came from the villages.METHODS:Questionnaires were filled up,and physical examinations were conducted in all the elderly. MAIN OUTCOME MEASURES: The 6 common diseases were observed among the elderly: Hyperplasia of prostate, abnormal audition, cartaract ,osteoarthritis, fracture and constipation.RESULTS:Totally 1434 elderly entered the stage of result analysis.①The incidence of hyperplasia of prostate in the males from the urban areas was lower than that in the rural areas (61.4％,65.7％, P ＜ 0.01 ). ② The incidence of abnormal audition among the elderly from the urban areas was lower than that in the rural areas (53.9％,64.7％,P ＜ 0.01).③ The incidence of cataract of the two eyes in the elderly from the urban areas was higher than that in the rural areas(46.4％,44.4％, P ＜ 0.01). ④ The incidence of osteoarthropathy in the elderly from the urban areas was higher than that in the rural areas (24.4％,14.9, P ＜ 0.01). ⑤ The incidence of fracture in the elderly from the urban areas was higher than that in the rural areas (14.2％,9.1％, P ＜ 0.05). ⑥ The incidence of constipation of the elderly in urban areas was lower than that in the rural areas(18.2％,23.0％,P ＜ 0.05).CONCLUSION:There is a difference between the elderly from the urban areas and the rural areas in hyperplasis of prostate,abnormal audition, cataract, osteoarthropathy, fracture and constipation, and this is related with the economy, nutrition, medical conditions and cultural traits.Therefore, attention should be attached to nutrition and health knowledge of the elderly and prevention and treatment of common diseases among the elderly people.

It was a single-center cross-sectional study to investigate the association of serum C3 level with blood pressure and estimated glomerular filtration rate (eGFR) in patients with idiopathic membranous nephropathy (IMN). The clinical and pathological data of 98 patients with IMN diagnosed by renal biopsy in the Department of Nephrology of East Hospital Affiliated to Tongji University from August 1, 2018 to October 31, 2023 were retrospectively analyzed. The demographic characteristics, serum complement C3 and other clinical data were compared between the non-hypertension group ( n=37) and hypertension group ( n=61). Pearson or Spearman correlation analysis method was used to analyze the correlation between serum C3 and eGFR in IMN patients and IMN patients with hypertension. Multiple linear regression analysis was used to analyze the related factors of eGFR in IMN patients with chronic kidney disease stage 1-3 in hypertension group. The results showed that compared with the non-hypertension group, the patients in hypertension group were older, and had higher levels of serum creatinine, cystatin C, urinary microalbumin to creatinine ratio, serum C3 and C4, and lower eGFR (all P<0.05). The correlation analysis showed that there was no correlation between serum C3 level and eGFR in IMN patients ( r=0.118, P=0.247). However, serum C3 level was positively correlated with eGFR in IMN patients with hypertension ( r=0.325, P=0.011). Multiple linear regression analysis showed that eGFR was negatively correlated with age ( β=-0.328, P=0.013), and positively correlated with serum C3 level ( β=0.228, P=0.048). The study shows that serum C3 level in hypertension group is higher than that in non-hypertension group in IMN patients. Moreover, serum C3 is positively correlated with eGFR.

OBJECTIVETo compare the effects of high, middle and low doses of enalapril in preventing left ventricular remodeling (LVRM) after acute myocardial infarction (AMI) in rats, especially evaluating the efficacy of low dose enalapril.

METHODSAMI was induced by ligating the left coronary artery in 149 female SD rats. 48 hours after the procedure, the 97 surviving rats were randomized to one of the following four groups: (1) AMI controls (n = 24), (2) high-dose (10 mg x kg(-1) x d(-1), n = 25), (3) middle-dose (1 mg x kg(-1) x d(-1), n = 23), and (4) low-dose (0.1 mg x kg(-1) x d(-1), n = 25) enalapril groups. In addition, sham-operated (n = 13) and normal rats (n = 10) were randomly selected to serve as non-infarction controls. Enalapril was delivered by direct gastric gavage. After 4 weeks of therapy, hemodynamic studies were performed, then the rat hearts were fixed with 10% formalin and pathology analysis was performed. Exclusive of the dead rats and those with MI size < 35% or > 55%, complete experimental data were obtained from 67 rats, which were comprised of (1) AMI controls (n = 13), (2) high-dose enalapril (n = 13), (3) middle-dose enalapril (n = 12), (4) low-dose enalapril (n = 12), (5) sham-operated (n = 8) and (6) normal (n = 9) groups.

RESULTSThere were no significant differences among the four AMI groups in infarction size (all P > 0.05). Compared with the sham-operated group, the left ventricular (LV) end diastolic pressure (LVEDP), volume (LVV), absolute and relative weight (LVAW, LVRW) in AMI group were all significantly increased (all P < 0.001), while maximum LV pressure rising and dropping rates (+/- dp/dt) and their corrected values by LV systolic pressure (+/- dp/dt/LVSP) were all significantly reduced in the AMI control group (P < 0.01 - 0.001), indicating LVRM occurred and LV systolic and diastolic functions were impaired. Compared with the AMI group, LVEDP, LVV, LVAW and LVRW were all significantly decreased in the three enalapril groups (control P < 0.001), with the reduction of LVEDP, LVV and LVAW being more significant in high-dose than in low-dose enalapril groups (all P < 0.05), and the +/- dp/dt/LVSP were significantly increased only in the high and middle-dose enalapril groups (P < 0.01).

CONCLUSIONSHigh, middle and low doses of enalapril were all effective in preventing LVRM after AMI in the rat, with low dose enalapril being effective and high dose superior. As for LV functional improvement, only high and middle-dose enalapril were effective.

Angiotensin-Converting Enzyme Inhibitors ; administration & dosage ; pharmacology ; Animals ; Dose-Response Relationship, Drug ; Enalapril ; administration & dosage ; pharmacology ; Female ; Myocardial Infarction ; physiopathology ; Rats ; Rats, Sprague-Dawley ; Ventricular Remodeling ; drug effects