1.Stem cells transplantation for experimental colitis in rats
Yuyuan LI ; Yuqiang NIE ; Jieying LAI ; Jinliang DUAN ; Yaming WEI ; Yanlei DU ; Weihong SHA ; Yongjian ZHOU
Chinese Journal of Digestion 2008;28(6):364-368
Objective To investigate the effect of transplantation of allogeneic bone marrow hematopoietic cells(HCs)and mesenchymal stem cells(MSCs)on experimental colitis(EC)in rats.Methods The HCs and MSCs obtained from SD male rats were cultured and expanded in vitro.In experiment 1 and 2 groups,HCs were labeled with bromodeoxyuridine(BrdU)and MSCs were obtained using the tube wall attach technique,respectively.Seventy-two female rats were infused with trinitrobenzene sulfonic acid(TNBS)to induce EC models.After 24 hours,HC or MSC suspensions were injected into the rats in experimental 1(n=18)and 2(n=18)groups via caudal veins,respectively.Control animals were injected with isotonic saline.The whole colon was removed on day 7,14 and 21 after transplantation and examined histopathologically.BrdU labeled HCs were tested with immunohistochemical staining and MSCs were detected for sex-determining gene(sry)by PCR.Results EC models were successfully established.The HCs or MSCs grew rapidly in the culture suspension.On day 7,14 and 21 after transplantation,the BrdU immunoreactive cells were detected in the colon(6/6),and the positive expression of the sry gene was found in 1/6,2/6 and 3/6,respectively.No positive labeled cell was found in controls.There was no significant improvement in histopathological scores on the colon in two experimental groups compared with the controls.Conclusions Allogeneic HCs and MSCs may localize in the colon of EC models.The ability of localization is higher in HCs than MSCs.The transplantation of HCs and MSCs can not obviously improve histopathologically.
2.Meta-analysis of insertion/deletion genetic variation of ACE gene and onset riskof type 2 diabetic nephropathy in Chinese population
Chunhua BEI ; Ying ZHANG ; Linyuan QIN ; Lin YANG ; Jieying DUAN ; Nian LIU ; Hongping YU ; Xiangyuan YU
Chongqing Medicine 2017;46(24):3362-3365
Objective To systematically assess the relation between angiotensin-I converting enzyme(ACE) gene insertion/deletion (I/D) variation and type 2 diabetic nephropathy (T2DN) onset risk among Chinese population.Methods The related literatures were retrieved from the China National Knowledge Infrastructure (CNKI) and Wanfang Data until June 1st,2016.The RevMan 5.0 was used to conduct the statistical analysis.The merge OR value and corresponding 95% confidence interval(95%CI) were used to assess ACE gene I/D polymorphism and T2DN onset risk.Results Totally 29 papers with 4 357 subjects were included according to the inclusion and exclusion standard,including 2 208 cases of DN and 2 149 cases of T2DM without DN.Meta analysis showed that compared with ACE gene I/D polymorphism I allele,D allele could significantly increase the risk of T2DM patients suffering from DN,the OR value and corresponding 95%CI were 1.44(1.25,1.66);the gene analysis showed that ACE gene I/D polymorphism loci were significantly correlated with DN onset risk in the Asian population.The corresponding relative onset risk OR and 95%CI were 1.42(1.15,1.76) and 1.75(1.46,2.10) in the dominant and recessive genetic model.The Begg′s test showed that the included data had no obvious publication bias existence.Conclusion ACE gene I/D polymorphism is closely correlated with the onset risk of T2DN,and D allele might be a risk genetic factor for DN occurrence in the patients with T2DM.
3.Establishment and application of Real-time quantitative PCR for human Metapneumovirus detection
Jieying ZHOU ; Yingbiao LIN ; Youde CAO ; Zhaojun DUAN
Chinese Journal of Experimental and Clinical Virology 2016;30(2):216-219
Objective To establish and evaluate a Metapneumovirus real-time quantitative reverse transcription polymerase chain reaction (RT-PCR),detect clinical specimens and explore the clinical prevalence characteristics of Metapneumovirus-infected children.Methods The primers and probes which targeted the conserved genes F were designed,RNA standards were prepared to establish a standard curve,the sensitivity,specificity and reproducibility had been tested.222 lower respiratory clinical specimens were collected from children in Lanzhou area with ARI.Metapneumovirus and co-infection viruses were detected simultaneously;further Metapneumovirus related epidemiology was studied.Results Metapneumovirus linear detection range was 10-10s copies/μl,the lowest detection limit was 10 copies/μl,the correlation coefficient was 1,the amplification efficiency was 91.62%,the CV of Ct value was less than 2%.Take conventional PCR product sequence results as reference,real-time quantitative RT-PCR sensitivity and specificity were 100% and 96.17%.Metapneumovirus detection rate were 9.46%,13 cases for boys (5.86%),8 case for girls (3.60%).The detection rate of spring,summer,autumn and winter were 15.71%,0%,5.08%,11.11%.There were no significant differences between the Metapneumovirus viral load and mixed infection or the types of disease,clinical symptoms.Conclusions Metapneumovirus realtime quantitative RT-PCR has been confirmed as a sensitive and specific method.Metapneumovirus was an important agent of children respiratory tract infection in Lanzhou region.We should take the long-term systematic surveillance seriously.
4.Real-time fluorescent quantitative PCR assay for detection of human rhinovirus
Jieying ZHOU ; Yaping SUN ; Haiyan CAO ; Zhiping XIE ; Zhaojun DUAN ; Youde CAO
Chinese Journal of Experimental and Clinical Virology 2014;28(2):129-131
Objective To establish the method for detecting human Rhinovirus (HRV) subtypes A,B,and C by reverse real-time fluorescent quantitative polymerase chain reaction (RT-PCR) and to assay HRV nasopharyngeal samples of children with acute respiratory tract infections.Methods The specific primers and Taqman probes of targeted HRV gene were designed,RNA standards were prepared to establish a standard curve,the sensitivity,specificity and reproducibility has been tested,222 clinical respiratory specimens were retrospectively detected.Results The linear ranges of human Rhinovirus A,B,and C subtypes were 109-10copies/μl,109-10 copies/μl,109-103copies/μl respectively,correlation coefficients were 0.999,0.997,0.999.Variation within the group was less than 3%.The detection rate was 27.02% by RT-PCR,the sensitivity and specificity were 100% and 98.18%.Conclusion The RT-PCR method can simultaneously detect rhinovirus subtypes A,B,and C with good sensitivity,specificity and reproducibility.
5.Epidemiological characteristics of viral agents in 1 092 children with acute lower respiratory diseases in Changsha
Jieying ZHOU ; Yaping SUN ; Yingbiao LIN ; Zhaojun DUAN ; Youde CAO ; Zhiping XIE
Chinese Journal of Experimental and Clinical Virology 2021;35(4):433-436
Objective:To explore the pathogenic spectrum and epidemiological characteristics of respiratory viruses in children with acute lower respiratory tract infection in Changsha, and provide scientific basis for disease prevention and treatment.Methods:A total of 1 092 respiratory tract specimens of children were collected and 12 respiratory viruses were detected by real-time quantitative transcription polymerase chain reaction.Results:Among the samples from 1 092 cases, those from 437 cases (40%) were positive for respiratory syncytial virus (RSV), 337 cases (30.9%) were positive for parainfluenza virus-3 (PIV-3), 263 cases (24.1%) were positive for human bocavirus (HBOV) and 228 cases (20.8%) were positive for adenovirus (ADV). The detection rates of boys and girls were 82.26% and 83.42%. The infection rate of RSV was higher in the group ≤ 6 months of age, the infection rates of PIV-3 and HBOV ≤2 years old were higher, and the infection rate of ADV was higher in the group between 6 months to 5 years old. The detection rates of virus infection of spring, summer, autumn and winter were 90.48%, 83.50%, 62.26% and 82.80%, respectively, there were significant differences among them.Conclusions:The main viruses in children with acute lower respiratory tract infection in Changsha were RSV and PIV-3. Mixed infections were common. Children under 2 years of age were more likely to get acute lower respiratory infections. Viruses had seasonal trends and peaked in winter and spring.