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

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.

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

An HIV-1 cell-cell fusion system was developed to screen HIV-1 entry inhibitors that block cell-cell fusion. In this system, the pEGFP-Tat plasmid was constructed and cotransfected into effector cells (HEK-293T) with HIV-1 envelope plasmid. TZM-bl cell, a genetically engineered cell line that expresses CD4, CXCR4, CCR5 as well as Tat-inducible β-galactosidase and luciferase reporter gene, was used as target cell. Thus, the co-culture of target cells and effector cells allows the cell fusion via Env and the activity of the fusion inhibitor can be quantified by measuring the reporter protein expression. The experimental parameters were optimized and 11 anti-HIV-1 agents including CCR5 antagonist maraviroc, reverse transcription inhibitor zidovudine (AZT) and integrase inhibitor raltegravir were tested. The result showed that the system exhibited high specificity and sensitivity. Two of eight tested anti-HIV-1 agents were found to block the cell-cell fusion. The system is suitable for efficient screening of HIV-1 cell-cell fusion inhibitors.

Decision-making aid for cancer patients is of great significance in the diagnosis and treatment for diseases. Breast cancer is one of the most common malignant tumors in women all over the world, and breast cancer patients have become the main target population for decision-aided research. Application of decision-making assistance for patients in Western countries has developed to a certain extent, while relevant research in China is still at the early stage. There are kinds of intervention forms for patients' decision aids, including traditional brochures and videos, decision aids systems, decision coaching, multidisciplinary breast cancer teams, etc. The tools for decision-making quality evaluation include the patients' awareness for decision-making, participation, decision-making conflict, decision-making satisfaction, decision-making regret, which can provide important guidance for the application of decision-making aid treatment in breast cancer patients in the future.
Breast Neoplasms/therapy* ; China ; Decision Making ; Emotions ; Female ; Humans ; Personal Satisfaction