1.Analysis of HCMV gN genotypes in patients with HCMV-HIV co-infection
Junling YU ; Jianjun WU ; Zhongwang HU ; Yanhua LEI ; Guolan LI ; Wenchang ZHANG ; Jun ZHAO ; Junling ZHANG ; Lin GAN ; Li YU ; Jason CHEN ; Mingli WANG
Chinese Journal of Microbiology and Immunology 2015;(4):258-264
Objective To analyze the distribution of various genotypes of human cytomegalovirus glycoprotein N ( HCMV gN) in patients with HIV infection; to investigate the effects of HCMV-HIV co-in-fection on disease progression and the relationships between HCMV gN genotypes and disease progression. Methods Patients with active HCMV infection were screened out from 359 patients with HIV infection by using the pp65 antigenemia assay.The genes encoding HCMV gN ( UL73 ) were amplified by nested PCR ( nPCR) .The amplicons were digested by restriction enzymes including MboⅠ, ScaⅠ and SalⅠ.Then, the restricted fragment length polymorphisms were further analyzed on 4%agarose gel.The relationships be-tween HCMV genotypes and the morbidity and mortality of acquired immune deficiency syndrome ( AIDS ) were investigated via a prospective study.Results Among the 359 patients with HIV infection, 28 subjects were positive for the HCMV pp65 antigenemia assay.The HCMV gN genotypes in 20 patients with active HCMV infection were distributed as: gN-3a (4/20, 20%), gN-1 (4/20, 20%), gN-4d (1/20, 5%), gN-4b (1/20, 5%) and mixed infection (10/20, 50%).Patients with HCMV-HIV co-infection were more likely to develop AIDS during the follow-up period (RR=9.78).Patients harboring HCMV gN-1 and gN-4 genotypes would seem likely to have 4.6 times of chance leading to AIDS-associated death than those harbo-ring other HCMV gN genotypes.Conclusion HCMV infection ( especially gN-1 and gN-4 genotypes) might accelerate the progression of HIV infection.
2.Preparation and identification of the polyclonal antibody against the 3D polymerase of foot and mouth disease virus
Yu ZHANG ; Yonglu WANG ; Yongguang ZHANG ; Yuzhen FANG ; Li PAN ; Shoutian JIANG ; Jianliang LU ; Likuan LIU ; Zhongwang ZHANG ; Shugang ZHANG ; Zhengfeng LI ; Jinxin DU
Chinese Journal of Zoonoses 2010;(1):6-9
To prepare the polyclonal antibody against the 3D polymerase of foot and mouth disease virus (FMDV), the 3D polymerase gene of this virus was amplified by PCR and doubly digested with BamH I and Nde I. Then, it were cloned into expression vector pET-30a(+) to obtain the recombinant plasmid pET-3D and this plasmid was transformed to E.coli BL21(DE3) with induction by IPTG.The target protein was identified and purified with SDS+PAGE, and the inclusion bodies were extracted. The purified target protein was used as antigen to immunize New Zealand rabbits to prepare the polyclonal antibody against 3D polymerase of FMDV, which was then characterized by indirect ELISA and Western blotting. As demonstrated by SDS-PAGE, the target protein with a molecular weight at 46 ku was expressed. The polyclonal antibody showed high affinity and obvious specificity and its titer was above 1:8 000. This polyclonal antibody may lay a foundation for the further studies on the biological functions and epitopes of the 3D polymerase of FMDV.
3.Comparative analysis of different states of polarized BV 2 cells by GC-MS com-bined with metabonomic technology
Yan WANG ; Zhongwang YU ; Si CHEN ; Ling LI ; Zhenyu ZHU
Journal of Pharmaceutical Practice 2015;(3):226-230
Objective To analyze the different metabolites of the classical activated (M1) ,alternatively activated (M2) and resting BV2 cells by metabolomics method .Methods The mRNAs of several potential biomarkers were determined by real-time PCR analyses to confirm the state of BV2 cells .Static GC-MS combined with metabolomics technology was used to analyze the metabolic changes .Results There were 15 biomarkers identified between the M1 group and the resting group ,and 15 biomarkers were found in the M2 group and the resting group .Conclusion The present study provides an effective way to reveal the mechanism of the polarization of BV 2 cell ,and it might provide a theoretical basis to prevent or treat the neurodegen-erative diseases .
4.Epidemiological investigation of a school COVID-19 outbreak in Shanghai
Zhongwang WANG ; Yan XIE ; Yu ZHANG ; Lingna KONG ; Yihui ZHENG ; Yuyan ZHANG
Shanghai Journal of Preventive Medicine 2023;35(11):1068-1073
ObjectiveTo describe a COVID-19 outbreak due to SARS-CoV-2 Omicron variant in a school and provide suggestions for COVID-19 prevention and control. MethodsData on the COVID-19 outbreak in a school in Putuo District of Shanghai were collected from November 14 to December 20, 2022. Epidemiological characteristics, incidence rate of secondary cases and response measures were described and analyzed. ResultsA total of 27 COVID-19 cases were identified infected with SARS-CoV-2 Omicron BA.5.2 variant, including 14 students (51.9%) and 13 family members / teachers living with those students (48.1%). The first case occurred on November 14, with peak incidence during November 16-18. The median generation interval of the second generation cases was 2 (2,3) days. The index case (case 1, a student) had a clear contact history outside the school, which was found through routine examination in key populations in the school. Immediate management was conducted after the notification. All the second generation cases were exposed students in the same class with case 1, which were identified during the quarantine, whereas the third generation cases were their family members/teachers living with the second generation cases. The incidence rate within the class and school were 36.8% and 3.0%, respectively. No further social transmission was found outside the school. ConclusionEarly detection, multi-sectoral collaboration, prompt control and quarantine measures are effective in containing SARS-CoV-2 transmission. Health promotion, surveillance, ventilation and prophylactic disinfection should be reinforced in schools, office buildings and other gathering places.
5. Pinocembrin Promotes OPC Differentiation and Remyelination via the mTOR Signaling Pathway
Qi SHAO ; Ming ZHAO ; Wenwen PEI ; Yingyan PU ; Mingdong LIU ; Weili LIU ; Zhongwang YU ; Kefu CHEN ; Hong LIU ; Li CAO ; Ming ZHAO ; Benqiang DENG ; Ming ZHAO ; Kefu CHEN
Neuroscience Bulletin 2021;37(9):1314-1324
The exacerbation of progressive multiple sclerosis (MS) is closely associated with obstruction of the differentiation of oligodendrocyte progenitor cells (OPCs). To discover novel therapeutic compounds for enhancing remyelination by endogenous OPCs, we screened for myelin basic protein expression using cultured rat OPCs and a library of small-molecule compounds. One of the most effective drugs was pinocembrin, which remarkably promoted OPC differentiation and maturation without affecting cell proliferation and survival. Based on these in vitro effects, we further assessed the therapeutic effects of pinocembrin in animal models of demyelinating diseases. We demonstrated that pinocembrin significantly ameliorated the progression of experimental autoimmune encephalomyelitis (EAE) and enhanced the repair of demyelination in lysolectin-induced lesions. Further studies indicated that pinocembrin increased the phosphorylation level of mammalian target of rapamycin (mTOR). Taken together, our results demonstrated that pinocembrin promotes OPC differentiation and remyelination through the phosphorylated mTOR pathway, and suggest a novel therapeutic prospect for this natural flavonoid product in treating demyelinating diseases.
6.Pinocembrin Promotes OPC Differentiation and Remyelination via the mTOR Signaling Pathway.
Qi SHAO ; Ming ZHAO ; Wenwen PEI ; Yingyan PU ; Mingdong LIU ; Weili LIU ; Zhongwang YU ; Kefu CHEN ; Hong LIU ; Benqiang DENG ; Li CAO
Neuroscience Bulletin 2021;37(9):1314-1324
The exacerbation of progressive multiple sclerosis (MS) is closely associated with obstruction of the differentiation of oligodendrocyte progenitor cells (OPCs). To discover novel therapeutic compounds for enhancing remyelination by endogenous OPCs, we screened for myelin basic protein expression using cultured rat OPCs and a library of small-molecule compounds. One of the most effective drugs was pinocembrin, which remarkably promoted OPC differentiation and maturation without affecting cell proliferation and survival. Based on these in vitro effects, we further assessed the therapeutic effects of pinocembrin in animal models of demyelinating diseases. We demonstrated that pinocembrin significantly ameliorated the progression of experimental autoimmune encephalomyelitis (EAE) and enhanced the repair of demyelination in lysolectin-induced lesions. Further studies indicated that pinocembrin increased the phosphorylation level of mammalian target of rapamycin (mTOR). Taken together, our results demonstrated that pinocembrin promotes OPC differentiation and remyelination through the phosphorylated mTOR pathway, and suggest a novel therapeutic prospect for this natural flavonoid product in treating demyelinating diseases.
Animals
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Cell Differentiation
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Flavanones
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Mice
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Mice, Inbred C57BL
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Myelin Sheath/metabolism*
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Oligodendroglia/metabolism*
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Rats
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Remyelination
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Signal Transduction
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TOR Serine-Threonine Kinases/metabolism*