Objective To observe the onset of vascular smooth muscle cell (VSMC) senescence induced by tert-butyl hydroperoxide (t-BHP) and the intervention effect of dehydroepiandrosterone (DHEA). Methods The VSMCs were divided into four groups: blank control group, t-BHP group (incubated with 80 μmol/L t-BHP for 72 h), 10 nmol/L DHEA intervention group (pretreated with 10 nmol/L DHEA 30 min before t-BHP) and 100 nmol/L DHEA intervention group (pretreated with 100nmol/L DHEA 30 min before t-BHP). Two ageing markers of ageing associated β-galactosidase activity and cell proliferation activity were adopted as main indexes. β-galactosidase activity was measured with immunocytochemical method and cell proliferation activity was measured with flowcytometry. Results After continuous treatment with 80 mmol/L t-BHP for 72 h, the ratios of G0/G1 phase cells and SA-β-galactosidase staining positive cells increased as compared with blank controlgroup [(89.4±3.4)% vs. (49.5±5.5)%, (3.5±1.2)% vs. (75.3±4.3)%], which indicated that VSMCs senescence were successfully induced by t-BHP. While the above changes were smaller in 100 nmol/L DHEA intervention group than in t-BHP group. Conclusions With ageing,accumulation of damage produced by reactive oxygen species may be an important mechanism causing the onset of VSMCs senescence. DHEA may be able to retard the progression of VSMCs senescence through antioxidant effect.

At present ,the mechanism of highly pathogenic avian influenza H5N1 virus causing human infection or death is still not fully clear .In order to better understand the pathogenesis of the disease ,the rhesus macaques were infected with H5N1 virus (AF148678/ACGoose/Guangdong/11961H5N1) .We analyzed the clinical symptoms ,characteristics of the virus invades body ,pathological changes ,and immune response to discuss the pathogenesis of viral pneumonia induced by H 5N1 virus infection from the early time to the recovery time .The rhesus macaques were infected with H5N1 virus through nasal .Clinical signs were assessed daily ,and major organs and blood were collected for detection of blood routine analysis ,viruses were isola-ted and titrated from organs ,and pathologic and immunohistochemical were also conducted .As a result ,the rhesus macaques in-fected with H5N1 virus experienced fever ,dyspnea ,and anorexia .The respiratory tract was the major target of the virus and the virus could not replicate in organs outside the respiratory tract .Positive staining cells by immunohistochemistry were bronchial epithelial cells and alveolar macrophages .Rhesus macaques experienced temporary severe pneumonia after 1-3 days ,mainly be-cause of neutrophils infiltration ;gradual recovery 6 days later ,mainly with macrophage infiltration ;lung tissue presented recov-ery state after 14 days ,mainly with T lymphocytes infiltration .Finally ,we concluded that the predilection of the H 5N1 virus to infect the lower airway suggests that it may be a limiting factor in human-to-human transmissibility of the H5N1 virus .The pathogenesis may include virus invasion ,replication and immune injury .

The systematic position of Fritillaria hupehensis has been in dispute. Phylogentic analyses were conducted on sequences of ITS, rpl16, matK sequences for species of F. hupehensis and allies. Lilium davidii was designed as outgroup. The analyses were performed using MP and ML methods. Conclusions could be achieved as follow. The topologies of MP and ML are consistent. The samples of F. hepehensis from different places form a supported clade with a strong bootstrap. And then form a strongly supported clade with F. anhuiensis, F. monantha. The results suggests that although F. hupehensis has a closet relation with the two ones, it exists some difference.
DNA, Plant ; chemistry ; genetics ; DNA, Ribosomal Spacer ; genetics ; Endoribonucleases ; genetics ; Fritillaria ; classification ; genetics ; Molecular Sequence Data ; Nucleotidyltransferases ; genetics ; Phylogeny ; Plant Leaves ; genetics ; Ribosomal Proteins ; genetics ; Sequence Analysis, DNA ; Species Specificity

Objective To evaluate the effects of infective necrosis (IN) on prognosis in moderately severe or severe acute pancreatitis (AP).Methods According to the revision of Atlanta classification,from January 2001 to January 2015,admitted patients with moderately severe or severe AP were retrospectively analyzed.According to whether with the presence of persistent organ failure (POF) and / or IN,the patients were divided into four groups:group one with weither IN nor POF,group two with IN but without POF,group three with POF but without IN,group four with both IN and POF.The differences in disease severity and prognosis among groups were compared.Logistic regression and Cox proportional hazard regression model were used to analyze the effect of IN on prognosis.Results A total of 375 moderately severe or severe AP patients were enrolled.There were 211,43,90 and 31 patients in group one,two,three and four,respectively.A total of 121 (32.3％) patients with POF,74 (19.7％) patients with IN,and death in 63 (16.8％) patients.The mortality rate in patients with IN was 32.4％ (24/74),and which was 13.0％(39/301) in patients without IN.The mortality rates of group one,two,three and four were 1.9％(4/211),11.6％(5/43),38.9％(35/90) and 61.3％(19/31),respectively;mortality rate was in a trend of increasing,and the difference was statistically significant (x2 =109.672,P＜0.01).Both IN (OR=8.24,95％CI2.09 to 32.46) andPOF (OR=8.31,95％ CI2.48 to 27.87)were independent risk factors of mortality of AP patients (both P＜0.01).Both IN (OR=2.04,95 ％CI 1.19 to 3.48,0.002) and POF (OR=5.25,95％CI 2.36 to 11.65) also were independent risk factors of shortened survival time of AP patients (both P＜0.01).Conclusions IN is an independent risk factor of disease severity and poor prognosis in AP.The prognosis is the worst in AP patients with both POF and IN.

Objective To investigate the inhibition effect of Ki67 AS-ODN on pr ostate carcinoma PC-3 cells,and its possible synergism existing in combination of Ki67 AS-ODN and paclitaxel.Methods Ki67 AS-ODN were transfected into PC-3 cells by lipofectamine. Cell proliferation was measured by CCK8 method,and Ki6 7 mRNA expression was detected by RT-PCR. The synergetic effect of Ki67 AS-ODN combined with paclitaxel was evaluated by Zhengjun Jin Q method. Results AS-OD N at the concentration of 31.25 nmol /L can significantly inhibit PC-3 cells pr oliferation(P

Objective To express,purify and identify recombinant hepatitis E virus(HEV) pb166-GST fusion protein using GST gene fusion system and investigate its potential role in researching Hepatitis E diagnostic antigen field.Methods The recombinant E.coli BL21 performed by our own laboratory was used to induce the HEV pb166-GST expression with IPTG.The products were purified by BD Biosience GST purifying system.The specific expression was identified by SDS-PAGE and Western blot.The experiment conditions and results were described and analysed.Results The resolved HEV pb166-GST fusion protein on SDS-PAGE showed a major band at position of 43 kD.The expressed proteins had a single expected band after purify and the protein was recognized by anti-GST antibody on PVDF membrane.Conclusion The recombinant HEV pb166-GST fusion protein is expressed in recombinant E.coli BL21 efficiently in this way,and might be used as a candidate for diagnostic antigen of HEV.

OBJECTIVETo observe the regulating effect of hepatitis C virus (HCV) envelop (E) 2 gene immunization-induced immune responses by adenovirus mediated interleukin 12 (IL-12).

METHODSHCV E2 protein was expressed and purified from NIH 3T3 and then used as an antigen to detect antibodies against HCV E2. With 51Cr release, SP2/0 expressing HCV E2 was used as target cell to detect specific cytotoxic T lymphocytes (CTL) response; adenovirus recombined IL-12 was propagated by 293 cell. HCV E2 recombinant and adenovirus recombined IL-12 were injected into the quadriceps femoris muscles and abdominal cavities of 6-8 weeks old BALB/C mice. Sera were collected at 2, 3, and 4 weeks and detected for antibodies for E2. Spleen cells isolated at 4 weeks were analyzed for specific CTL response.

RESULTSIt was found that expression of IL-12 at an undetectable level did enhance HCV E2 gene immunization-induced CTL activity and there was no effect on its hormonal immune response.

CONCLUSIONUsing adenovirus to express interleukin 12 was helpful for regulation of HCV E2 gene immunization-induced immune response. Combined HCV E2 and IL-12 can render a strong anti-HCV CTL activity and may be of use in the development of HCV gene vaccine in the future.

Adenoviridae ; physiology ; Interleukin-12 ; biosynthesis ; genetics ; T-Lymphocytes, Cytotoxic ; immunology ; Viral Envelope Proteins ; genetics ; immunology

Bacteria ; isolation & purification ; Cross Infection ; microbiology ; Drug Resistance, Bacterial ; Female ; Humans ; Infant, Newborn ; Intensive Care Units, Neonatal ; Male

Objective To understand the prevalence of urinary incontinence(UI) and its related factors so as to develop a three-tier program for prevention of the disease.Methods A cross-sectional study was carried out in two townships of Jixian county,Tianjin,during July to November 2007.A total of 743 people aged 60 years and over were selected under cluster sampling,and all information were collected with a standardized structured questionnaire by face-to-face interview.All the data were analyzed with multivariate logistic regression method to explore the related factors for UI in the elderly.Results The overall prevalence of UI was 33.38 percent among people aged 60 years and over in two townships.Risk factors flor UI in men would include older age (OR=1.39).occupation(OR=5.00),awareness of UI(OR=1.91),having in chronic respiratory diseases(OR=2.23),prostate(OR=11.47),neurological(OR=11.76),or motor systems (OR=2.48).while protective factors would include high educational level(taking primary school or below as control group),OR for the junior middle school group appeared to be 0.35,for senior middle school group it was 0.77.and ofundergraduate group it was 0.53.Risk factors for UI in women would include older age(OR=1.31),constipation(OR=1.46),awareness of UI(OR=1.94),increased bodv nlass index(when normal weight group served as control group,OR in the overweight group was 1.03 and in the obesity group OR was 1.54),suffering from chronic respiratory diseases(OR=4.84).diabetes mellitus(OR=2.36),or motor system diseases(OR=1.37),more gravidity(OR=1.03),more parity(OR=1.02),suffering from perinea laceration(OR=1.72) and wound infection during delivery(OR=1.65),while protective factors would include physical exercises(OR=0.64).Conclusion Prevalence of UI was higher among the elderly people in rural areas of Jixian county,Tianjin.UI in the elderly might have been influenced by various factors which suggesting the intervention strategy should be targeted at those related factors as well as focusing on primary prevention.

Objective To understand the prevalence of urinary incontinence(UI)and its severity in rural elderly people,as well as to investigate the awareness on UI in the elderly and health-care service seeking behavior.Methods A cross-sectional study was carried out in two townships of Jixian county,Tianjin.A total of 743 people aged 60 years and over were selected under cluster sampling method.All the information was collected with a standardized structured questionnaire by face-to-face interview.Prevalence,severity,the awareness on UI in the elderly and their utilization of health-care service for its diagnosis and treatment were analyzed.Results Prevalence of UI was 33.38% among people aged 60 years and over in two townships of Jixian,higher in females than in males(43.15% vs.22.75%,χ~2=34.70,P＜0.0001).The prevalence rates of UI in 60-age group,65-age group,70-age group,75-age group,80-age group,85-95 age group were 28.64%,32.12%,34.08%,35.45%,47.76%,30.00%,respectively,and increased with age (for trend χ~2=219,P=0.029).Stress urinary incontinence(SUI)and urge urinary incontinence(UUI)were mild,while mixed urinary incontinence(MUI)were mainly moderate.The differences of severity of SUI,UUI,MUI between men and women did not show statistical significance(all P＞0.05).In 743 elderly people,more than half of the respondents had never heard of UI(50.20%,373/743)and only 170(22.88%)elderly people considered UI as a disease.630(84.79%)and 665(89.50%)elderly people in our research group did not know that such condition was curable and preventable.In 248 elderly people with UI,only 12(4.84%)of them ever seeking community health-care services in the health-care centers or hospitals.For the ones who did visit the centers,the purpose was only to seek for drug treatment.Conclusion Prevalence of UI appeared to be high among the elderly people in rural areas of Jixian county.Most of the elderly people were lack of knowledge about UI that hindered them from seeking for diagnosis and treatment in the clinics,plus the treatment program for UI was not standardized.Knowledge on UI and health care seeking behavior should be popularized and strengthened among elderly people living in the rural areas.