1.Peroxisome proliferator-actived receptor-? ligand troglitazone induces apoptosis in renal cell carcinoma
Fengguang YANG ; Zhiwen ZHANG ; Dianqi XIN ; Changjin SHI ; Xueqing WU ; Wujiang LIU ; Yinglu GUO ; Jieping WU ;
Journal of Peking University(Health Sciences) 2004;0(02):-
Objective: To investigate the expression of peroxisome proliferator actived receptor ? (PPAR ?)and the inducement of apoptosis by PPAR ? ligand in renal cell carcinoma(RCC) derived cell lines.Methods:RT-PCR and Western blot analysis were performed to determined the expression of PPAR ? mRNA and protein in two RCC derived cell lines(786 O and A498) and two normal kidney(NK) derived cell lines(HK 2 and HMCC). Two RCC cell lines were treated with 50 ?mol/L troglitazoned for and evaluated for the effects of antidiabetic thiazolidinediones (TZDs) on the cells apoptosis by fluorescence microscopy and DNA ladder assay.The mutative expressions of Bcl 2 and Bax before and after TZDs treatment were also performed by western blot analysis. Results: The expression of PPAR ? was observed to be stronger in 786 O and A498 cells than in HK 2 and HMCC cells by RT-PCR and Western blot analysis. Treated with 50 ?mol/L troglitazone (for 48 h) it induced typical apoatosis in 786 O and A498 cells. After treatment, a decrease in Bcl 2 expression in RCC cells was observed by Western blot analysis,and the expression of Bax,however,was up regulated.Conclusion: The results reveal that troglitazone has the tumor suppressive effect on RCC cells. High affinity PPAR ? ligands (TZDs) may be the candidates for a novel approach to the treatment of this refractory neoplasm.
2.Establishment and application of LAMP-Taqman detection method of porcine pseudorabies virus
Yu LI ; Lei SHI ; Guoqiang SHI ; Yinglu ZHANG ; Zhenguo DONG
Chinese Journal of Veterinary Science 2024;44(10):2110-2115
A LAMP-Taqman rapid detection system for porcine pseudorabies virus(PRV)was de-veloped based on LAMP and quantitative PCR.LAMP primers were designed for PRV conserved sequences,and the loop primer modified by the fluorescent quenching group was used as the Taq-man probe.The composition optimization,specificity,sensitivity and repeatability of the LAMP-Taqman system were tested using positive samples and recombinant plasmid as templates.Thirty-eight samples of pork swabs were tested with the commercial LAMP detection kit in parallel to verify the actual detection effect of the LAMP-Taqman detection system.The results showed that the optimal final concentration of each component was as follows:PRV-FIP/BIP 0.8 μmol/L,Bst DNA polymerase 0.7 U/μL,Taq DNA polymerase 0.24 U/μL,dNTPs 1.6 mmol/L,MgSO47.2 mmol/L.This system had good specificity and did not cross-react with other virus samples.The linear correlation coefficient of gradient samples was 0.995,the coefficient of variation of repeatable tests was less than 3.000%,and the minimum detection limit could reach 2.81 ×102 copies/μL.The test results of the actual swab samples were consistent with the commercial isothermal fluores-cence detection reagents.In conclusion,the PRV detection system established by LAMP-Taqman method in this study is specific,sensitive,stable and accurate,and is a reliable technical method suitable for the accurate detection of porcine PRV.
3.Whole genome sequence analysis of coxsackievirus A group 4 isolates in Anhui province in 2020
Yinuo LIU ; Yinglu GE ; Lingkang YANG ; Yong SUN ; Yonglin SHI
Chinese Journal of Experimental and Clinical Virology 2024;38(3):269-278
Objective:To understand the genome-wide sequence variation and molecular evolution of coxsackievirus A4 (CV-A4) strain in Anhui province, so as to provide a theoretical basis for the pathogenic monitoring and scientific prevention and treatment of hand, foot and mouth disease in the future.Methods:Five CVA4 isolates of 2020 were sequenced by first-generation sequencing method. MEGA11.0 was used to construct a phylogenetic tree based on VP1 region for 5 CV-A4 isolates, 32 CV-A4 strains and Enterovirus A71(EV-A71) prototype strain BrCr, and the isolates and enterovirus A (EV-A) prototype strains based on P1, P2 and P3 regions respectively, and DNAStar was used for amino acid sequence comparison in VP1 region. BioEdit7.2 was used for amino acid displacement entropy analysis and amino acid site entropy mapping. SimPlot3.5 and RDP4 were used for recombination analysis of CV-A4 isolate and EV-A prototype representative strains, and DnaSP6 software was used for selection pressure analysis of isolates and reference representative strains.Results:The phylogenetic tree showed that the isolates belonged to the C2 subtype, which belonged to the same clade as the CV-A4 strain circulating in Chinese mainland, and the amino acid sequence homology of the C2 subtype branch was 97.3%-100%, and the isolates had 6 amino acid variation sites compared with the prototype. The selection pressure analysis showed that the CV-A4 strain of the C2 subtype was affected by negative selection pressure, and there were 25 mutagenic sites in the amino acid sequence in the coding region of the displacement entropy analysis, accounting for 1.14%, and the evolution of the strain mainly depended on recombination. Recombination analysis showed that the isolates recombined with a variety of EV-A prototype strains in the P2 and P3 regions, and the recombination section with the CV-A5 prototype strain was longer, especially in the 3A-3C section, and P1 was a relatively conserved region.Conclusions:CV-A4 has frequent recombination events with other EV-A prototype strains in P2 and P3, and the molecular evolution of CV-A4 in Anhui should continue to be closely monitored.
4.In vitro effect of zilongjin on prostate cancer cell line LNCaP.
Xue-song LI ; Yun-yan LIANG ; Dai-shu WANG ; Xianhe XIE ; Changjin SHI ; Jie CHEN ; Guofeng SUN ; Liqun ZHOU ; Zhaoying XUE ; Yangun NA ; Yinglu GUO
Chinese Journal of Integrated Traditional and Western Medicine 2004;24(7):621-624
OBJECTIVETo investigate the effect of Zilongjin (ZLJ) on human androgen-dependent type of prostate cancer cell line LNCaP.
METHODSMTT assay, flow cytometry and fluorescence microscopy were used to observe the effect of ZLJ in anti-proliferation, cell cycle arresting and apoptosis induction. RT-PCR was used to examine the effect of ZLJ on expressions of prostate marker gene (PSA), androgen receptor (AR), apoptosis related genes (bcl-2 and bax), and Western blot assay was used to detect the effect on protein expression of bcl-2 and bax.
RESULTSZLJ could cause apparent inhibition on proliferation, induce G0/G1 phase arresting and apoptosis in time- and dose-dependent manner on LNCaP cells. The concentration for inhibiting cell growth by 50% (IC50) in 72 hrs was 0.79 mg/ml. ZLJ could down-regulate the expression of PSA, AR, bcl-2 genes and lower bcl-2 protein expression, but showed ineffective on bax protein expression.
CONCLUSIONZLJ displays its anti-tumor effects by way of inhibiting the cell proliferation, arresting the G0/G1 phase, inducing apoptosis, down-regulating PSA, AR, bcl-2 gene expression and lowering bcl-2 protein expressions.
Antineoplastic Agents, Phytogenic ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Male ; Neoplasms, Hormone-Dependent ; metabolism ; pathology ; Prostate-Specific Antigen ; biosynthesis ; genetics ; Prostatic Neoplasms ; metabolism ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; biosynthesis ; genetics ; Receptors, Androgen ; biosynthesis ; genetics
5. Genetic analysis of norovirus strains detected from sporadic cases in Anhui province, 2016-2017
Yuan YUAN ; Yonglin SHI ; Yong SUN ; Wanfu HU ; Weiwei LI ; Yinglu GE
Chinese Journal of Experimental and Clinical Virology 2019;33(2):136-141
Objective:
To analyze the genotype diversity and phylogenetic characterization of norovirus(NoV) in patients with diarrhea from Anhui province.
Methods:
NoV positive fecal specimens from sentinel hospitals were collected from January, 2016 to December, 2017. The samples were detected by Real-time fluorescent quantitative PCR. Positive samples were of randomly selected and amplified by RT-PCR and the products were sequenced. Phylogenetic tree was constructed by the Neighbor-Joining method based on partial VP1 gene regions of NoV to perform phylogenetic analysis.
Results:
A total of 263 NoV positive samples were genotyped, of which 239 belonged to genogroup II, 24 belonged to genogroup I. Fifty-five positive samples were successfully sequenced. There were 6 NoV GII genotypes, which included GII.2, 3, 4/Sydney_2012, 13, 17 and 21, while NV GII.17 and GII.4 were the dominant genotypes from 2016 to 2017. The predominant genotype was GII.4/Sydney 2012 (47.27%, 26/55), followed by GII.17 (23.64%, 13/55) and GII.2 (14.55%, 8/55). Phylogenetic tree showed that 26 strains belonged to genotype GII.4/Sydney 2012, NoV. The nucleotide homology among the 26 VP1 genes was 97.8% to 100%. Analysis of the partial VP1 genes of 26 strains showed that it shared the highest homology of 98.9% with the strain of GII.4Sydney2012 (GenBank ID: KU720515). However, the prevailing genotype in the Anhui province has shifted on two separate occasions, the GII.17 strain was dominant in 2016, and the GII.4/Sydney 2012 strain was dominant in 2017.
Conclusions
NoV GII was the major pathogen causing sporadic diarrhea in Anhui province during from 2016 to 2017, the genotypes are widely distributed, and shifted into the two predominant strains.
6.Detection and analysis of clinical specimens of COVID-19 in Anhui province
Qingqing CHEN ; Jun HE ; Junling YUN ; Yonglin SHI ; Weiwei LI ; Yuan YUAN ; Yinglu GE ; Nan SA ; Meng WANG ; Zhuhui ZHANG ; Yong SUN
Chinese Journal of Experimental and Clinical Virology 2020;34(4):382-384
Objective:To examine 2019 novel coronavirus (2019-nCoV) RNA in clinical specimens of COVID-19 patients in Anhui province, and provide evidence for laboratory diagnosis of COVID-19 and risk assessment of clinical specimens.Methods:ORF1ab gene and N gene of 2019-nCoV were detected by real-time fluorescence RT-PCR in 466 clinical specimens of 197 COVID-19 cases. Chi-square test was used to analyze the differences in positive rates of specimens with clinical classification and time of onset.Results:The positive rates of 2019-nCoV in throat swab, sputum, serum, blood sample were 88.83%, 94.67%, 6.78% and 5.08%. The positive rate for 2019-nCoV RNA in throat swabs and sputum differed significantly ( χ2=8.994, P=0.003) in common cases during 7 days after illness onset. Conclusions:The positive rate of RNA in sputum was higher than throat swabs. 2019-nCoV RNA was detected in serum and blood specimens of COVID-19 cases. There was a risk of serum and blood specimens for transmission of COVID-19.
7.Early outcome of transapical transcatheter aortic valve replacement for aortic insufficiency
Yinglu SHI ; Chengxin ZHANG ; Zhixiang GUO ; Wenhui GONG ; Shenglin GE
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2023;30(08):1116-1120
Objective To assess the early outcome of transapical transcatheter aortic valve replacement (TAVR) for patients with aortic insufficiency. Methods The patients with aortic valvular disease who underwent transapical TAVR from October 2020 to October 2022 in the Department of Cardiac and Vascular Surgery, the First Affiliated Hospital of Anhui Medical University were enrolled in the current retrospective study. The patients with aortic stenosis were assembled in a group A, and the patients with aortic insufficiency were assembled in a group B. The improvements of heart function and complications were assessed for the two groups. Results A total of 56 patients were enrolled, including 32 males and 24 females with an average age of 73.34±5.10 years. There were 31 patients in the group A and 25 patients in the group B. There was no statistical difference between the two groups in the age, gender, height, weight, hypertension, coronary artery disease, peripheral vascular disease, chronic obstructive pulmonary disease, renal disorder or classification of heart function (P>0.05). Also, there was still no statistical difference in the rate of permanent peacemaker implants, emergent open surgery, valve re-implants, or perivalvular leakage (P>0.05). After TAVR, the left ventricular diastolic diameter, left ventricular ejection fraction, complicated moderated mitral and tricuspid regurgitation were significantly improved in both groups compared with preoperative findings (P<0.05); however, there was no statistical difference in these parameters between groups (P>0.05). Conclusion Interventional valve (J-Valve) in the treatment of patients with aortic insufficiency through transapical TAVR significantly improves cardiac function and reduces functional valve regurgitation.
8.Epidemiological characteristics and pathogen spectrum changes of hand-foot-mouth disease in Anhui province from 2015 to 2022
Wanwan Ma ; Lei Gong ; Yongkang Xiao ; Yonglin Shi ; Yinglu Ge ; Sai Hou ; Yong Sun ; Guoping Chen ; Yuanfang Sun ; Hongyuan Wei ; Shuang Nie ; Sun Liang ; Yong Sun
Acta Universitatis Medicinalis Anhui 2023;58(10):1763-1768
Objective :
To analyze the epidemiological characteristics and pathogen spectrum of hand,foot mouth disease (HFMD) in Anhui province from 2015 to 2022,and to provide scientific evidence for prevention and control measures of HFMD.
Methods :
The surveillance data of hand,foot and mouth disease in Anhui province from 2015 to 2022 were analyzed by descriptive epidemiology. Real-time PCR was used to detect and classify HFMD samples.
Results :
A total of 650 590 HFMD cases were reported in Anhui province from 2015 to 2022,including 1 406 se- vere cases and 17 deaths.The annual reported incidence was 131. 45 /100 000.The epidemic features of“low incidence in odd years and high incidence in even years”were presented from 2015 to 2019.The incidence showed a continuous decline from 2020 to 2022.The monthly distribution showed the characteristics of bimodal epidemic,and the main peak was not obvious in 2020.Hefei,Fuyang,Bozhou,Chuzhou and Suzhou ranked the top five cities in terms of cumulative incidence.The age of onset was mainly distributed in children aged 5 years and below,accounting for 89. 26% of the total cases.The male to female ratio was 1. 48 ∶ 1.A total of 28 657 laboratory-confirmed cases had been reported from 2015 to 2022.EV71 cases accounted for 10. 57% ,Cox A16 cases accounted for 24. 90% ,and other enterovirus cases accounted for 64. 53%.The dominant pathogens showed dynamic changes in different years.Since 2018,the proportion of EV71 decreased significantly,and the proportion of other enteroviruses gradually increased to become the dominant pathogens.Among other enteroviruses,Cox A6 strain was dominant (80. 48% ) .
Conclusion
This study suggests that the prevention and control of HFMD in Anhui province should be paid more attention from April to July and from October to December.The focus areas are the cities in northern Anhui and Hefei where the floating population is large.The focus of prevention and control is on children aged 5 years and below.Other enteroviruses have become the dominant pathogens of hand-foot-mouth disease in Anhui province,Cox A6 strain is dominant.