中华微生物学和免疫学杂志最新文献

{"title":"Bioinformatics prediction of molecular mechanism and intervention drugs of SARS-related immune injury and their significance for COVID-19 treatment","authors":"Haomin Zhang, Haoran Chen, Yakun Yang, Ximeng Chen, Jundong Zhang, B. Guo, P. Zhi, Zhuo-guo Li, Geliang Liu, Boxuan Yang, Xiao-hua Chi, Yixing Wang, Xue-chun Lu","doi":"10.3760/CMA.J.CN112309-20200215-00064","DOIUrl":"https://doi.org/10.3760/CMA.J.CN112309-20200215-00064","url":null,"abstract":"Objective \u0000To investigate the omics mechanism of SARS-related immune injury and predict targeted therapeutic drugs through clinical bioinformatics analysis of the transcriptome data of SARS virus in order to provide reference for clinical treatment of COVID-19. \u0000 \u0000 \u0000Methods \u0000The transcriptome data of SARA virus were collected from the Gene Expression Oibus (GEO) and used to screen differential genes. Enrichment analysis and protein interaction analysis were performed to investigate the mechanism of immune damage associated with SARS. A platform of epigenetics in precision medicine (EpiMed) was established to predict potential therapeutic drugs. \u0000 \u0000 \u0000Results \u0000The mechanism of SARS-related immune injury was complex, involving affecting the function of immune cells through signaling pathways such as Toll-like receptors, increasing cytokines in plasma through Th17 signaling pathway and inducing autoimmune responses after autoantibodies were generated by molecules such as IL-6, NF-κB, and TNF. Drugs such as Chuanqiong and Etanercept might have therapeutic effects on SARS-related immune damage. \u0000 \u0000 \u0000Conclusions \u0000SARS virus could cause abnormal expression of many immune-related molecules and signaling pathways. Drugs such as Chuanqiong and Etanercept might have therapeutic effects on SARS-related immune damage. This study might provide reference for clinical treatment of COVID-19. \u0000 \u0000 \u0000Key words: \u0000SARS virus; COVID-19; Immune injury; Clinical bioinformatics; Drug prediction","PeriodicalId":10089,"journal":{"name":"Chinese journal of microbiology and immunology","volume":"40 1","pages":"165-173"},"PeriodicalIF":0.0,"publicationDate":"2020-03-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"43522041","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Immunoreactivity and diagnostic value of Mycobacterium tuberculosis PstS1 and HspX","authors":"Na Li, Machao Li, T. Xiao, Yuhan Yan, Xiaoqin Li, Hai-can Liu, K. Wan","doi":"10.3760/CMA.J.CN112309-20191104-00364","DOIUrl":"https://doi.org/10.3760/CMA.J.CN112309-20191104-00364","url":null,"abstract":"Objective \u0000To evaluate the humoral and cellular immunoreactivity of recombinant Mycobacterium tuberculosis (M.tuberculosis) PstS1 and HspX protein antigens in order to provide reference for immunodiagnosis of tuberculosis and screening of candidates for vaccine antigens. \u0000 \u0000 \u0000Methods \u0000Purified recombinant M. tuberculosis PstS1 and HspX proteins were obtained using molecular cloning expression and Ni2+ affinity chromatography. Blood samples and epidemiological data of healthy individuals and patients with M. tuberculosis infection were collected. Specific IgG antibodies and IFN-γ-producing antigen-specific T cells were respectively detected by enzyme-linked immunosorbent assay (ELISA) and enzyme-linked immunospot assay (ELISPOT) with the recombinant proteins used as antigens. The humoral and cellular immunoreactivity of the recombinant PstS1 and HspX proteins were assessed with statistical analysis of data. \u0000 \u0000 \u0000Results \u0000Both the recombinant PstS1 and HspX proteins could induce the secretion of IFN-γ by more specific effector T cells in patient with M. tuberculosis infection, and the differences between the infection and healthy control groups were statistically significant (P<0.05). The specificity and sensitivity of the recombinant PstS1 and HspX as the diagnostic antigens of ELISPOT were 92.11% (35/38) and 65.96% (31/47), and 68.42% (26/38) and 91.49% (43/47), respectively. The two proteins also possessed some humoral immunoreactivity, but statistically significant difference was only observed in the HspX-specific antibody level between the two groups (P<0.05). \u0000 \u0000 \u0000Conclusions \u0000Both the recombinant PstS1 and HspX proteins had good cellular immunoreactivity and were the immunodominant antigens of cellular immunity. They performed well in cellular immunodiagnosis and were good potential candidate antigens for anti-tuberculosis vaccines. \u0000 \u0000 \u0000Key words: \u0000Mycobacterium tuberculosis; PstS1; HspX; Immunodominant antigens","PeriodicalId":10089,"journal":{"name":"Chinese journal of microbiology and immunology","volume":"40 1","pages":"192-197"},"PeriodicalIF":0.0,"publicationDate":"2020-03-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46397075","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Construction and expression of chimeric antigen receptors targeting epidermal growth factor receptor (EGFR) and programmed cell death ligand-1(PD-L1)","authors":"Shuping Li, Xiao-jue Wang, Bin Yang, Helin Wang, Zhuohong Yan, L. Yi, Panjian Wei, Xin Jin, J. Hao, Hongtao Zhang","doi":"10.3760/CMA.J.CN112309-20191105-00365","DOIUrl":"https://doi.org/10.3760/CMA.J.CN112309-20191105-00365","url":null,"abstract":"Objective \u0000To construct an expression system of lentivirus vector encoding epidermal growth factor receptor-specific chimeric antigen receptor (EGFR-CAR) and programmed cell death ligand-1 (PD-L1) antibody. \u0000 \u0000 \u0000Methods \u0000Human PD-L1-Fc protein was used to immunize BALB/c mice. Cell-fusion and subcloning were performed to screen stable hybridoma strains with high secretion of PD-L1-specific antibodies, which were identified by both ELISA and Western blot. The activity of the antibodies in blocking the binding of programmed cell death-1 (PD-1) to PD-L1 was determined by fluorescence-activated cell sorting (FACS). Antibody affinity was analyzed by Fortebio Octet96. A single-chain variable fragment (scFv) was further constructed after antibody full-length sequencing and humanization using CDR grafting method. Meanwhile, the genes encoding the light and heavy chain variable regions (VL and VH) were cloned from a hybridoma secreting antibody against human EGFR by 5′ RACE technology to construct scFv gene. The expression of scFv was confirmed using pcDNA3.1 vector. EGFR-CAR containing CD137 intracellular function domain and PD-L1-scFv was ligated using 2A gene. The synthetic single molecule was cloned into pLVX-EF1a-IRES-ZsGreen1 lentivirus expression vector, and then transfected into 293T cells using Lenti-X Packaging Single Shots (VSV-G) to prepare infectious virus. Expression of CAR on cell surface and the soluble form of PD-L1-scFv in the supernatant of transfected 293V cells were detected by FACS and ELISA. \u0000 \u0000 \u0000Results \u0000A PD-L1 antibody named 11E3 with high ligand-receptor blocking performance was obtained. The humanized antibody showed a stable affinity (2.67×10-10 mol/L) after directly grafting the mouse CDRs (CDR1, CDR2 and CDR3) to human frameworks. EGFR-scFv was effectively expressed in a form of Fc-fusion. Secretory CAR (CTZ0431-1) and membrane CAR (CTZ0431-2) expression plasmids were constructed using lentivirus vector containing EGFR-CAR and PD-L1-scFv. The infection efficiency in 293V cells was around 10%. EGFR-scFv on the cell membranes and PD-L1-scfv in the culture supernatants were detected after 293V cells were infected with CTZ0431-1. EGFR-scFv and PD-L1-scfv were expressed on the cell membranes of 293V cells infected with CTZ0431-2. The expression rate of CAR in LV-CART46407-1-transfected activated T cells was 39.3%. \u0000 \u0000 \u0000Conclusions \u0000The lentivirus vectors co-expressing EGFR-CAR with moderate binding affinity and PD-L1-scFv with high binding affinity were successful constructed, which provided an essential tool for investing EGFR- and PD-L1 double targeted CAR-T cell therapy against solid tumor. \u0000 \u0000 \u0000Key words: \u0000EGFR-CAR; PD-L1-scFv; Membrane expression; Secretory expression; Synthetic biotechnology","PeriodicalId":10089,"journal":{"name":"Chinese journal of microbiology and immunology","volume":"40 1","pages":"198-205"},"PeriodicalIF":0.0,"publicationDate":"2020-03-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"47876057","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Research of simvastatin-induced autophagy in inhibiting recombinant human metapneumovirus replication","authors":"Pan Zhang, Suhua Chen, Hui Yang, Tingting Wu, Yao Zhao","doi":"10.3760/CMA.J.CN112309-20190923-00306","DOIUrl":"https://doi.org/10.3760/CMA.J.CN112309-20190923-00306","url":null,"abstract":"Objective \u0000To study whether simvastatin could inhibit viral replication during human metapneumovirus (hMPV) infection. \u0000 \u0000 \u0000Methods \u0000Human bronchial epithelial cells (16HBE) were infected with hMPV and then treated with or without simvastatin. Real-time quantitative PCR (qPCR) and Western blot were used to detect virus titers and the activation of autophagy and related pathways. BALB/c mice were infected with hMPV and then treated with simvastatin through intragastric administration. Pathological changes in lung tissues were observed. Changes in viral loads and the activation of autophagy and related pathways in proteins and RNA extracted from lung tissues were detected. \u0000 \u0000 \u0000Results \u0000The in vitro experiment showed that the hMPV+ simvastatin group had decreased virus titer and enhanced autophagy than the hMPV group. The AKT/mTOR pathway in the hMPV+ simvastatin group was inhibited, which was verified by a further experiment using rapamycin, a specific inhibitor of AKT/mTOR pathway. The in vivo experiment showed that the virus titer in the hMPV+ simvastatin group was lower than that in the hMPV group, but there was no significant difference in the activation of autophagy. The AKT/mTOR pathway was down-regulated in the hMPV+ simvastatin group. HE staining revealed that obvious pathological changes were observed in the hMPV group, but the condition was improved after simvastatin intervention. \u0000 \u0000 \u0000Conclusions \u0000Simvastatin can inhibit the replication of hMPV, which is associated with the activation of autophagy induced by AKT/mTOR pathway. \u0000 \u0000 \u0000Key words: \u0000Human metapneumovirus; Simvastatin; Autophagy; AKT/mTOR","PeriodicalId":10089,"journal":{"name":"Chinese journal of microbiology and immunology","volume":"40 1","pages":"185-191"},"PeriodicalIF":0.0,"publicationDate":"2020-03-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"48839788","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Expression and antibody preparation of recombinant truncated glycoprotein of Guertu virus","authors":"Abula Ayipairi, S. Shěn, Jìngyuàn Zhāng, Xijia Liu, Yijie Li, F. Dèng, Yujiang Zhang, Sùróng Sūn","doi":"10.3760/CMA.J.CN112309-20190727-00231","DOIUrl":"https://doi.org/10.3760/CMA.J.CN112309-20190727-00231","url":null,"abstract":"Objective \u0000To express truncated glycoprotein (Gn, Gn1, Gn2, Gn3, Gc1 and Gc2) of Guertu virus (GTV) in Escherichia coli (E.coli) cells, and prepare polyclonal antibodies against recombinant proteins Gn-His, Gc1-His and Gc2-His after purification. \u0000 \u0000 \u0000Methods \u0000Gene fragments encoding Gn, Gn1, Gn2, Gn3, Gc1 and Gc2 of GTV DXM strain were amplified by RT-PCR, and cloned into the prokaryotic expression vector pET-32a (+ ) to construct recombinant expression plasmids. The transformed E. coli BL21(DE3) strains carrying expression plasmids were induced by IPTG to express target proteins, which were identified by SDS-PAGE. Recombinant proteins Gn-His, Gc1-His and Gc2-His were purified by nickel affinity chromatography and detected by Western blot using GTV-positive sheep serum for analysis of their antigenicity. New Zealand white rabbits were immunized with the purified recombinant proteins. The titers and specificity of serum antibodies were analyzed by ELISA. Meanwhile, eukaryotic expression vectors pcDNA3.1-Gn, pcDNA3.1-Gc1/Gc2 were constructed and transfected into mammalian Vero cells to evaluate the binding activity of rabbit polyclonal antibodies by indirect immunofluorescence method. The specific reactivity of serum antibodies to recombinant proteins was detected by Western blot. \u0000 \u0000 \u0000Results \u0000Restriction enzyme analysis and DNA sequencing confirmed that the recombinant expression vectors of pET-32a-Gn, pET-32a-Gn1/Gn2/Gn3, pET-32a-Gc1/Gc2, pcDNA3.1-Gn and pcDNA3.1-Gc1/Gc2 were constructed successfully. The relative molecular mass (Mr) of the expressed recombinant proteins Gn-His, Gn1/Gn2/Gn3-His, Gc1/Gc2-His were approximately 63.4×103, 37.1×103, 31.9×103, 30.8×103, 40×103 and 54.4×103, respectively. The recombinant proteins could be recognized by GTV-positive sheep serum. The titers of polyclonal antibodies against GTV Gn, Gc1 and Gc2 were 1∶409 600, 1∶204 800 and 1∶6 400, respectively. Indirect immunofluorescence assay and Western blot showed that the prepared rabbit polyclonal antibodies could specifically react with the proteins expressed in eukaryotic cells and the recombinant proteins. \u0000 \u0000 \u0000Conclusions \u0000The recombinant GTV glycoproteins Gn-His and Gc1/Gc2-His were efficiently expressed and purified and characterized with good immunity. The prepared polyclonal antibodies had high titers and good specificity. This study provided reference for further studying the biological function and detection methods of GTV glycoproteins and research on vaccines. \u0000 \u0000 \u0000Key words: \u0000Guertu Virus; Glycoprotein; Prokaryotic expression; Eukaryotic expression; Rabbit polyclonal antibody","PeriodicalId":10089,"journal":{"name":"Chinese journal of microbiology and immunology","volume":"40 1","pages":"178-184"},"PeriodicalIF":0.0,"publicationDate":"2020-03-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"45264265","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Progress in mucosal immune of female reproductive tract","authors":"Huihui Wang, Wenhui Qi, Xingshuo Li, A. Fan, Cha Han, F. Xue","doi":"10.3760/CMA.J.CN112309-20190710-00211","DOIUrl":"https://doi.org/10.3760/CMA.J.CN112309-20190710-00211","url":null,"abstract":"The mucosal immune system is the first line of defense against pathogens such as bacteria and viruses invading the bodies. The female reproductive tract mucosal immune system not only resists the invasion of pathogens through innate and adaptive immunity, but also contribute to successfully fertilization and pregnancy, thus maintaining the health of women′s reproductive system. The innate immunity of female genital tract involves the mechanical barrier of the mucosal epithelium, microbial barrier of commensal bacteria, immunological barrier of immune cells and their receptors, and adaptive immunity including B cell-mediated humoral immunity and T cell-mediated cellular immunity. Female genital mucosal immunity is not only involved in local inflammation, but may also have anti-tumor effects. Moreover, the female genital tract mucosal immune is also regulated by sex hormone to maintain a homeostasis of local microenvironment. Herein, this paper summarized recent progress in female genital mucosal immunity. \u0000 \u0000 \u0000Key words: \u0000Mucosal immune; Female reproductive tract; Innate immunity; Adaptive immunity; Commensal bacteria; Sex hormone","PeriodicalId":10089,"journal":{"name":"Chinese journal of microbiology and immunology","volume":"40 1","pages":"238-243"},"PeriodicalIF":0.0,"publicationDate":"2020-03-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46500355","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Whole genome sequencing of SARS-CoV-2 isolated in Guangdong Province and factors influencing the sequencing","authors":"L. Liang, Bosheng Li, Zhe Liu, Zhencui Li, Qianfang Guo, Yingchao Song, Xue Zhuang, L. Zou, Jianxiang Yu, Jie Wu","doi":"10.3760/CMA.J.CN112309-20200216-00065","DOIUrl":"https://doi.org/10.3760/CMA.J.CN112309-20200216-00065","url":null,"abstract":"Objective \u0000To obtain the genome sequences of SARS-CoV-2 in respiratory specimens in Guangdong Province with next-generation sequencing (NGS) and analyze the factors influencing sequencing. \u0000 \u0000 \u0000Methods \u0000Eight upper and lower respiratory tract specimens were collected from patients with SARS-CoV-2 infection in Guangdong Province in January 2020. RNA library construction was used to obtain the genome sequences of SARS-CoV-2. A bio-informatics software package (CLC Genomics Workbench 12.0) was used to analyze and compare the genomic sequences. \u0000 \u0000 \u0000Results \u0000Five SARS-CoV-2 genome sequences were obtained from the eight specimens and two were obtained from lower respiratory tract specimens. The nucleotide homology to SARS-CoV-2 was 97.74%-99.90%. The Ct values were lower, while the sequencing depth, coverage, relative abundance and genome integrity were higher in sequencing the SARS-CoV-2 in lower respiratory tract specimens. \u0000 \u0000 \u0000Conclusions \u0000The low Ct value of SARS-CoV-2 in the samples was good for sequencing. \u0000 \u0000 \u0000Key words: \u0000SARS-CoV-2; Next-generation sequencing; Influencing factor","PeriodicalId":10089,"journal":{"name":"Chinese journal of microbiology and immunology","volume":"40 1","pages":"174-177"},"PeriodicalIF":0.0,"publicationDate":"2020-03-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"45040232","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Role of peptidyl arginine deiminase 4 (PAD4) in anti-β2GP1/β2GP1 complex-mediated neutrophil extracellular trap formation","authors":"Bin Sun, Q. Cai, Si Cheng, Guo-ying Xu, Yongxin Chen, Hongxiang Xie","doi":"10.3760/CMA.J.CN112309-20190606-00178","DOIUrl":"https://doi.org/10.3760/CMA.J.CN112309-20190606-00178","url":null,"abstract":"Objective \u0000To investigate the role of peptidyl arginine deiminase 4 (PAD4) in anti-β2GP1/β2GP1 complex-induced formation of neutrophil extracellular trapping networks (NETs). \u0000 \u0000 \u0000Methods \u0000Peripheral blood neutrophils were isolated from healthy humans by density gradient centrifugation. PAD4 expression was detected by Western blot after the neutrophils were incubated with anti-β2GP1/β2GP1 complex (100 μg/ml) for a certain period of time. PAD4 inhibitor Cl-amidine (10 μmol/L) was used to pretreat neutrophils. Changes in the expression of citrullinated histone 3 (CitH3) at protein level and the relative content of myeloperoxidase (MPO)-DNA were detected by Western blot and ELISA, respectively. A mouse thrombus model of antiphospholipid syndrome (APS) was established by inferior vena cava stenosis. Intervention experiments were performed by intraperitoneal injection of Cl-amidine (50 mg/kg). The expression of CitH3 at protein level in plasma was detected by Western blot. The concentration of circulating free DNA (cf-DNA) in plasma was measured with fluorescent staining. Thrombus in inferior vena cava was collected and weighted to evaluate whether inhibiting the activity of PAD4 would suppress the APS-IgG-induced formation of NETs and thrombosis. Differences among groups were analyzed by t test or one-way analysis of variance (ANOVA). \u0000 \u0000 \u0000Results \u0000The expression of PAD4 induced by anti-β2GP1/β2GP1 complex was significantly down-regulated in the cytoplasm, but increased in the nucleus [(3.67±0.32) vs (1.47±0.19), t=10.22, P<0.05; (0.57±0.19) vs (2.97±0.31), t=11.49, P<0.05]. Cl-amidine significantly inhibited the anti-β2GP1/β2GP1 complex-induced expression of CitH3 protein by neutrophils [(2.46±0.47) vs (0.46±0.13), t=12.24, P<0.01], and reduced the MPO-DNA content in the culture supernatants [(4.09±0.94) vs (2.80±0.57), t=4.23, P<0.05]. In vivo, Cl-amidine significantly inhibited the expression of CitH3 protein [(3.97±0.56) vs (1.09±0.45), t=11.83, P<0.01] and decreased the content of cf-DNA [(2 685.0±735.8) vs (1 784.0±577.0), t=3.93, P<0.05] in plasma of APS mice. Compared with the experimental APS mice in the control group, the weight of thrombus in the APS mice pretreated with Cl-amidine was significantly reduced [(8.22±3.06) vs (4.89±1.90), t=2.27, P<0.05]. \u0000 \u0000 \u0000Conclusions \u0000PAD4 was involved in the formation of NETs induced by anti-β2GP1/β2GP1 complex, which might play an important role in APS thrombosis. \u0000 \u0000 \u0000Key words: \u0000Antiphospholipid syndrome; Anti-β2GP1/β2GP1 complex; Neutrophil extracellular traps; Peptidyl arginine deiminase 4","PeriodicalId":10089,"journal":{"name":"Chinese journal of microbiology and immunology","volume":"40 1","pages":"115-121"},"PeriodicalIF":0.0,"publicationDate":"2020-02-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"44758767","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Effects of sample processing modes on high-throughput sequencing of coronavirus whole genome/ 不同样本测序前处理方案对冠状病毒基因组高通量测序结果的影响","authors":"Jing Chen, Ya-Wei Zhang, P. Niu, Rojian Lu, N. Zhu, Hang Fan, W. Tan","doi":"10.3760/CMA.J.CN112309-20191011-00322","DOIUrl":"https://doi.org/10.3760/CMA.J.CN112309-20191011-00322","url":null,"abstract":"Objective: To study the effects of different pre-sequencing sample processing modes on the results of whole genome sequencing with high-throughput sequencing (HTS) by taking the largest RNA virus (human coronavirus, HCoV) as the representative. Methods: Cell-cultured human coronavirus HCoV-OC43 strains were used as the representative samples and divided into different groups based on pre-sequencing processing modes as follows: untreated group, DNase and RNase treatment before nucleic acid extraction group, DNase treatment after nucleic acid extraction group, and DNase and RNase treatment before nucleic acid extraction and DNase treatment after nucleic acid extraction group. Nucleic acid samples of each group were analyzed by direct RNA sequencing (without amplification) and DNA sequencing after sequence independent single primer amplification (SISPA), respectively. Results: No significant difference in viral genome coverage rates was observed between different groups. The highest genome coverage and sequencing accuracy were obtained in DNase treatment after nucleic acid extraction group by direct RNA sequencing, and the ratio of viral reads and the sequencing depth of each locus were effectively improved by SISPA amplification. Conclusions: This study provided an optimized technical strategy for whole genome sequencing of RNA viruses such as coronavirus.","PeriodicalId":10089,"journal":{"name":"Chinese journal of microbiology and immunology","volume":"2 1","pages":"103-109"},"PeriodicalIF":0.0,"publicationDate":"2020-02-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"45456558","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Regulation and mechanism of mannan-binding lectin (MBL) on adipogenic differentiation of 3T3-L1 preadipocytes","authors":"Yonghui Yang, Jingwen Yang, Zhixin Li, Yanwei Sun, Yimeng Yang, Wei Zhang, Fanping Wang, Lili Yu, Mingyong Wang","doi":"10.3760/CMA.J.CN112309-20191106-00370","DOIUrl":"https://doi.org/10.3760/CMA.J.CN112309-20191106-00370","url":null,"abstract":"Objective \u0000To investigate the regulatory effects and mechanism of mannan-binding lectin (MBL) on adipogenic differentiation of 3T3-L1 preadipocytes. \u0000 \u0000 \u0000Methods \u00003T3-L1 preadipocytes were induced to differentiate into adipocytes in vitro, and stimulated with different concentrations of MBL (0, 1, 10, 20 μg/ml). Firstly, changes in cell proliferation ability were detected by CCK-8. Then lipid accumulation was analyzed by Oil red O staining and intracellular triglyceride content determination. Further, the expression of adipogenic differentiation-related factors PPARγ and C/EBPα at protein and mRNA levels were detected by Western blot and qRT-PCR, respectively. Finally, Western blot was used to analyze the expression and phosphorylation of Akt, a signal molecule related to adipogenic differentiation. \u0000 \u0000 \u0000Results \u0000MBL at the concentrations of 0, 1, 10 and 20 μg/ml had no effect on the proliferation of 3T3-L1 preadipocytes. The level of triglyceride in MBL treatment groups decreased in a dose-dependent manner on 3 d after 3T3-L1 preadipocyte differentiation. Results of Oil red O staining showed that the number of lipid droplets in MBL treatment groups reduced significantly, and the absorbance values also decreased significantly in a concentration-dependent manner. Western blot and qRT-PCR results showed that the expression of PPARγ and C/EBPα at both protein and mRNA levels in MBL treatment groups decreased significantly in a dose-dependent manner, and the phosphorylation level of Akt was significantly down-regulated as well. \u0000 \u0000 \u0000Conclusions \u0000MBL regulates the adipogenic differentiation of 3T3-L1 preadipocytes via Akt signaling pathway. \u0000 \u0000 \u0000Key words: \u0000Mannan-binding lectin; 3T3-L1 preadipocyte; Akt; Adipogenic differentiation","PeriodicalId":10089,"journal":{"name":"Chinese journal of microbiology and immunology","volume":"40 1","pages":"122-128"},"PeriodicalIF":0.0,"publicationDate":"2020-02-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41974076","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}