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Objective@#To evaluate the effectiveness of AIDS intervention programs on men aged 50 or over and having had non-marital sexual behavior.@*Methods@#A community-based intervention/experimental and based on individual level study was adopted. Stratified sampling method was used. 12 townships/streets in Fuyang district of Hangzhou were identified as intervention or control group (six research sites each). All of the subjects in the township (street) were included. The inclusion criteria of study objects would include men aged 50 or older who reported having unmarried sex in the last year. Estimated sample size was 290, with each 145 in the intervention group and the control group. All the intervention group participants were provided with a total of 4 intervention-related items (knowledge and education on AIDS prevention, information radiation and behavioral change, broadcast expert lectures), every 3 months, for 12 month, the main evaluation indicators would include: incidence of non-marital sex and commercial sex in the last year, condom use when having non-marital sex in the last episode.@*Results@#A total of 312 subjects were recruited. 300 of them completed the baseline study while 284 of them completed the follow-up survey. Among the subjects who had undergone the baseline study, the average age was (65.58±7.89), 71.33% were married or cohabiting with someone, 52.00% having had primary school education. After the implementation of intervention programs, the incidence of non-marital sex dropped to 59.42% (82/138) and the incidence of commercial sex dropped from 79.73% (118/148) to 55.07% (76/138). Condom use rate in the last non-marital sexual contact increased from 19.59% (29/148) to 51.22% (42/82). In the control group, the incidence of non-marital sex in the year before dropped to 74.66% (109/146) and the incidence of commercial sex dropped from 91.45% (139/152) to 72.60% (106/146). Rates of condom use during the last non-marital sexual contact dropped from 32.89% (50/152) to 31.19% (34/109). Statistically, there were significant differences appeared between the two groups on the incidence of non-marital sex in the past year (χ2=7.48, P=0.008), the incidence of commercial sex in the last year (χ2=9.47, P=0.003) and the rate of condom use in the last sex experience (χ2=7.83, P=0.007).@*Conclusions@#Results from this intervention study showed that: in the intervention group, both the incidence rates of non-marital or commercial sex had reduced, together with the increase of condom use in non-marital sex in the last sexual experience. Intervention strategies that involving knowledge and education on AIDS prevention, information radiation and behavioral change, broadcasting lectures by experts etc. were all proved effective.
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Objective To investigate the G protein-coupled receptor kinase 2 (GRK 2) level in peripheral blood lymphocytes with cardiac func-tion in elderly patients with acute myocardial infarction. Methods This study enrolled 40 patients with acute ST-segment elevation myo-cardial infarction (STEMI) and 40 patients with unstable angina. All patients were 65 years or older. Cardiac function was evaluated by echocardiography, and the GRK 2 level in peripheral blood lymphocytes was measured. Patients with STEMI were followed up for 2 years. Results The GRK 2 level in peripheral blood lymphocytes was significantly higher in patients with STEMI than in patients with unstable angina, and was negatively correlated with left ventricular ejection fraction, cardiac output, stroke volume, and left ventricular fractional shortening. The GRK 2 level was significantly elevated in some patients with acute STEMI and poor cardiac function. Conclusions In-creased GRK 2 level in patients with acute STEMI may contribute to poor myocardial systolic function and myocardial remodeling. Meas-urement of the GRK 2 level in peripheral blood lymphocytes may assist in the evaluation of cardiac function and myocardial remodeling in elderly patients with acute STEMI.
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Objective To investigate the effects of ω-3 fish oil lipid emulsion via vein on the inflammatory response,immune and organ function in patients with severe acute pancreatitis.Methods A total of 53 patients with severe acute pancreatitis were randomized into conventional therapy plus fish oil group (FO group) and conventional therapy group (CON group).The patients in FO group were treat with ω-3 fish oil lipid emulsion (0.2 g · kg-1 · d-1,10％) based on conventional therapy for 14 days.The level of C-reactive protein (CRP),TG and TC were detected before treatment and at day 7 and day 14 after treatment.CD4+,CD4+/CD8+ and C3,C4 were also detected at day 1 and day 14 after treatment.At the same time,acute physiology and chronic health evaluation Ⅱ score (APACHE Ⅱ score),intra-abdominal pressure,negative fluid balance time,enteral nutrition start-time and ICU stay time were observed and recorded.Results Forty-five out of 53 patients were finally recruited into results statistics.The level of CD4+,CD4+/CD8+ and C3 at day 14 after treatment in FO groups improved significantly than that in the CON group (P ＜0.05).The levels of CRP,intra-abdominal pressure and APACHE Ⅱ score at day 7 and day 14in FO group descended more obviously than that in the CON group (P ＜ 0.05).The negative liquid balance time in FO group (3.55 ±0.86) days was obvious shorter than that in CON group (4.61 ± 1.12) days,while enteral nutrition start-time (3.86 ± 1.17) days was significantly earlier compared with CON group (5.30 ± 1.61) days (P ＜ 0.05),however ICU stay time and 28 days mortality rate had no significant difference between the two groups.Conclusions ω-3 fish oil lipid emulsion can decrease the inflammatory response and the negative liquid balance time,improve the immune function and restore bowel function in severe acute pancreatitis patients.Therefore,it maybe provide a new and effective means for severe acute pancreatitis.
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@#ObjectiveTo assess the safety of intramyocardium injection of sarcoplasmic reticulum Ca2+-ATPase (SERCA2a) to rescue chronic heart failure beagles. MethodsThe heart failure beagles were assessed with myocardium enzymology, cAMP content of myocardium, myocardial oxygen consumption, arrhythmia, systemic inflammation factors, and the function of liver and kidney after SERCA2a gene transferred. ResultsThere were no increase of cAMP content, no more arrhythmia, myocardial oxygen consumption, no apparente systemic inflammation, and no injures to hepatic and renal function. ConclusionOverexpression of SERCA2a by intramyocardium injection is relatively safe.
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BACKGROUND: There are still few effective methods to repair injured myocardium after myocardial failure and pathologically rebuild reverral myocardium. As a new therapy, normal myocytes and therapeutic gene to interfere injured myocardium have advantageous effects in improving heart function.OBJECTIVE: To observe the efficiency and stability of adenovirus-medicated gene transferred into different passages of bone marrow mesenchymal stem cell (MSC) and investigate the effect of MSC-based sarcoplasmic reticulum Ca2+ ATPase gene (SERCA2a) gene therapy for rats with chronic heart failure. To compare the effects of gene therapy, cell transplantation and MSC-based SERCA2a gene therapy for chronic heart failure. DESIGN: Randomized controlled study.SETTING: Department of Senile Angiocardiopathy, General Hospital of Chinese PLA; Department of Biochemistry, Beijing Medical University. MATERIALS: Male Sprague-Dawley (SD) rats with 4 weeks old, clean grade and weighing 45-50 g provided by the Animal Experimental Center, Peking Medical University were used as donators of bone marrow. Other female SD rats of 12 weeks old, clean grade and weighing 200-250 g were used as receptors of cell transplantation and gene therapy. Sry gene of Y chromosome in male rats was used to evaluate whether transplanted cells of donators lived in myocardium of receptor rats. Ad-SERCa2a and Ad-EGFP were constructed by Doctor Lu Xiao-chun; MSC in the 3rd and 8th generations was isolating cultured on its own. METHODS: The experiment was carried out in the Zhou CY Laboratory (BSL-2), Department of Biochemistry, Beijing Medical University from July 2004 to December 2005. Thirty female SD rats received ligation at the left coronary artery to make models with chronic cardiac failure following acute myocardial infarction. And then, 29 rats were randomly divided into four groups, including gene therapy group (n=7), MSC group (n=7), gene-modified MSC group (n=8) and control group (n=7). Rats in the four groups were given the interventions of SERCA2a gene, MSC transplantation, MSC+Ad/SERCa2a and empty adenoviral vector, respectively. MSCs were separated and cultured, and then Ad-SERCA2a-GFP was used to transfer MSC in the 3rd and 8th generations.MAIN OUTCOME MEASURES: Ad-SERCA2a-GFP transfection rate of MSC was measured by using flow cytometer. Before and at 14 and 21 days after treatment, cardiac function was evaluated by ultrasonic echocardiogram. Expression of cytokine Ⅷ was tested by immunohistochemical staining. SERCA2a gene and protein expression were evaluated by RT-PCR and Western blot respectively, as well as SERCA2a enzyme activity. RESULTS: ① Transfection rate: The infection efficiency of adenovirus-medicated gene into different passages of MSC was over 80%, and there was no difference between passage three (P3) MSC and P8 MSC (P > 0.05). ② Heart function: Left ventricle wall was thickened obviously in group MSC and group MSC+Ad/SERCa2a on the 21st day after treatment, while volume was shortened and gradually rounded. Compared to control group, ejection fraction (EF) and shortening fraction (FS) of group Ad-SERCa2a, group MSC and group MSC+Ad/SERCa2a were elevated significantly on the 14th day after therapy (P < 0.01). While the elevation values of EF and FS began to reduce in group Ad-SERCa2a on 14th day after therapy, it continued to increase in both group MSC and group MSC+Ad/SERCa2a (P < 0.01). Improvement rate of EF at 21 days after therapy (EF D21) increased in group MSC and group MSC+Ad/SERCa2a respectively, but decreased in group Ad-SERCa2a. Compared to group Ad-SERCa2a, peak systolic flow velocity of anterior wall and interventricular septum in group MSC+Ad/SERCa2a increased significantly on the 21st day after therapy, and peak diastolic flow velocity of anterior wall and interventricular septum elevated in group MSC+Ad/SERCa2a, too (P < 0.01). ③ SERCA2a gene, protein expression and enzyme activity in group MSC+Ad/SERCa2a were significantly stronger in group MSC and control group. Parts of MSC transplanted into scar zone expressed Ⅷ.CONCLUSION: ① MSC is an effective platform for the targeted delivery of therapeutic gene. It suggests that different passages of MSC from P3 MSC to P8 MSC are regarded as high-effectively gene vehicles. MSC-based SERCA2a gene therapy showed much strong and lasting beneficial effect on exhausted myocardium. ② Effect of MSC transplantation on improving heart function may be related to promoting vascular neogenesis.
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BACKGROUND:In chronic congestive heart failure (CHF),both protein expression and activity of sarcoplasmic reticulum (SR) Ca2+ ATPase (SERCA2a) are decreased, which results in abnormal regulation of calcium ion and systole and diastole dysfunction.OBJECTIVE:To observe the short-and middle-term therapeutic effects of adeno-associated viral gene transfer of SERCA2a in treatment of CHF in rats,and to investigate the possible mechanisms.DESIGN:Randomized-controlled design.SETTING:Department of Geriatric Cardiology and Department of Pathophysiology,General Hospital of Chinese PLA.METHODS:Male Sprague-Dawiey rats,were randomized into 4 groups:sham-operation group,CHF group,CHF+ green fluorescent protein (GFP) group and CHF+SERCA2a group. Gene transfer 10 and 30 days two time points were set in each group.CHF rat models were established by coarctation of abdominal aorta.At postoperative 18 to 20 weeks,the successful rats in CHF group were intraperitoneally injected with 500 μL aseptic normal saline,and those in the CHF+ GFP group and CHF+ SERCA2a group were injected with the same dose of Adeno-associated viral gene with GFP and SERCA2a gene respectively.MAIN OUTCOME MEASURES:①At 10 and 30 days after gene transfer,hemodynamic parameters,the SERCA2a protein expression was analyzed by Western Blot,and SERCA2a activity was carded out by modified Jones method.②The difference of proteome of hearts was detected by expression proteomics in between CHF+ SERCA2a group and CHF group at 30 days.③Electrophoretic separation and quantitation of cardiac myosin heavy chain(MHC) isoforrns were done at 30 days.RESULTS: ①In CHF group and CHF+GFP group, SERCA2a expression and activity were lower than those in sham-operation group.At 10 days after gene transfer.SERCA2a expression was increased by 80.7%and still lower than that of sham-operation group,and activity was also increased by 89.9%,but still lower than that of sham-operation group.And at 30 days,the SERCA2a protein expression and activity in the CHF+SERCA2a group reached to levels of sham-operation group. ②At day 10,LVSP and LV +dp/dtmax and LV-dp/dtmax were increased and LVEDP was decreased dramatically,but they still did not reach to the levels of sham-operation group.At day 30,the parameters were normalized back to control levels.There were no significant differences in each index between CHF+SERCA2a group and sham-operation group.③Overexpression of SERCA2a increased some energy metabolic enzymes of hearts at 30 days.④There were a downregulation of α-MHC and an upregulation of 3-MHC in failing hearts.Overexpression of SERCA2a increased the expression of α-MHC and decreased the expression of β-MHC to the levels of sham-operation group at 30 days.CONCLUSION: Adeno-associated viral gene transfer of SERCA2a can enhance SERCA2a functions, maintain calcium homeostasis,improve cardiac energy metabolism,and normalize the expression of MHC isoforms in CHF rats.As a result,the heart functions can be improved. So adeno-associated viral gene transfer of SERCA2a has good therapy effects on CHF rats in short and relatively long periods.
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BACKGROUND: The direct ratio is formed between the size of osseous defect around implant and the required time of complete repair. It is viewed that bone implantation is suggested if osseous defect is larger than 1 mm, which benefits the bone regeneration and early fixation of implant.OBJECTIVE: To compare the effects between coralline granules and hydroxyapatite (HA) during union after immediate implantation.DESIGN: Group observation and controlled experiment was designed.SETTING: Department of Stomatology Renmin Hospital of Wuhan University.MATERIALS: HA coated implant, HA granule, coralline granule and 3adult mongrel dogs.METHODS: The experiment was performed in Department of Stomatology, Renmin Hospital of Wuhan University from August 2002 to April 2003.Under anesthesia, 6 foramens were drilled on femurs of 3 dogs (3 foramens on each side) to result in osseous defect. Coralline granules were embedded in the osseous defect around the implant in all of proximal ends (coralline granule group, CG group), HA granules were embedded in the osseous defect around the implant in all of distal ends (HA group) and nothing was embedded in the defect around the implant in the center (the control). One dog was sacrificed under anesthesia on the 2nd, 3rd and 4th months after operation successively and the samples were collected from the implanted section in each group for X-ray examination and scanning electronic microscopic observation.MAIN OUTCOME MEASURES: X-ray examination on implanted sections and scanning electronic microscopic observation on samples in each the 4th month, it was observed that implants and osseous tissue were closely integrated in CG group and HA group and there was still partial osseous on samples in each group: on the 4th month, it was discovered that the regenerated osseous tissue was matured completely in CG group and few coralline granule residue was left. In HA group, the regenerated osseous tissue was matured completely, but there was still a large amount of HA granules left and the granules had not been absorbed obviously. In the control group, the space appeared partially in the implanted cervical region.CONCLUSION: The implantation of artificial bone is suggested if the osseous defect around immediate implant is larger than 1 mm. As the implanting materials, coralline granules and HA granules act on the most advantageous guide activity. Coralline granules can be degraded and substituted with osseous tissue,but HA granules cannot be absorbed, which affects osseous reconstruction.
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<p><b>OBJECTIVE</b>Constructing a plasmid containing the shRNA of luciferase to suppress the expression of luciferase in BHK-21 cell.</p><p><b>METHODS</b>A 334 bp human U6 snRNA promoter was amplified from human genomic DNA by PCR and ligated to a 21 bp reverse repeated motif of luciferase target sequence with 9 bp spacer and AAV plasmid pSNAV. The recombinant pSNAV/U6/Luc plasmid cotransfected with pMAMneoLuc or transfected luciferase cell line to detect the effect of luciferase expression separately.</p><p><b>RESULTS</b>pSNAV/U6/luc suppresses the luciferase expression from pMAMneoLuc by 50% and luciferase cell line by 70%.</p><p><b>CONCLUSIONS</b>The results showed that the short hairpin RNA of luciferase can efficiently suppress its expression in BHK-21.</p>
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Humans , Cell Line , Gene Expression , Genetic Vectors , Luciferases , Genetics , Plasmids , Genetics , Promoter Regions, Genetic , RNA , Genetics , TransfectionABSTRACT
Objective To investigate the therapeutic effects of recombinant adeno-associated virual gene serotype 1(rAAV1) mediated transfer of sarcoplasmic reticulum Ca2+ ATPase(SERCA2a) on beagle dogs with heart failure(HF).Methods The chronic HF model was reproduced in beagle dogs by giving rapid right ventricular pacing(230 beats/min) for 30 days.A reduced rate(180 beats/min)was continued for another 30 days.Sixteen beagle dogs were divided into four groups(4 each): control group,HF group,HF+EGFP group and HF+SERCA2a group.After rapid pacing for 30 days,rAAV1-EGFP(1?1012vg/ml) and rAAV1-SERCA2a(1?1012vg/ml) were respectively delivered via intramyocardial routes,while no treatment was given to the animals in both control and HF groups.At the end of the study,haemodynamics,echocardiography and the protein expression of SRCA2a were measured respectively.The apoptosis index of cardiac myocyte was evaluated by TUNEL.Bax expression was assessed by immunohistochemistry.Results After gene transfer of SERCA2a in HF beagle dogs for 30 days,the heart function was improved along with an increase in SERCA2a expression.Left ventricular systolic function was significantly increased,including the left ventricular systolic pressure(LVSP),left ventricular maximal rate of pressure rise(LV+dp/dtmax),left ventricular maximal rate of pressure decline(LV-dp/dtmax,P
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Objective To survey and analyze the etiology of pericardial effusion in different age patients. Methods The data of 450 patients who were diagnosed as pericardial effusion were studied retrospectively. The pathogenesis of pericardial effusion were analyzed and compared among the five age groups : juvenile group (0-19 years),young group (20-39 years),middle age group (40-59 years), aged group (60-79 years) and existed advanced age group (≥80 years). Results The different pathogenesis of pericardial effusion existed in different age groups. The first three causes are tumor, tuberculous, heart failure in all patients. As a pathogeny, cardiopulmonary insufficiency induced pericardial effusions constitute the majority (about 50%) in the advanced age group. Tumor, heart failure are the main causes in aged group. In middle age group, tumor and tuberculous are the most frequent pathogenesis, while the tuberculous is the principal causes in juvenile and young groups. Conclusions The rate of tumor, heart failure, lung infection raised along with the age increasing in the patients with pericardial effusion, meanwhile the cases of tuberculoses declined. Attention should be given to this tendency on the diagnosis and treatment of the patients with pericardial effusions.