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Applications and developments of gene therapy drug delivery systems for genetic diseases 被引量:4
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作者 Xiuhua Pan Hanitrarimalala Veroniaina +4 位作者 Nan Su Kang Sha Fenglin Jiang Zhenghong Wu Xiaole Qi 《Asian Journal of Pharmaceutical Sciences》 SCIE CAS 2021年第6期687-703,共17页
Genetic diseases seriously threaten human health and have always been one of the refractory conditions facing humanity.Currently,gene therapy drugs such as siRNA,shRNA,antisense oligonucleotide,CRISPR/Cas9 system,plas... Genetic diseases seriously threaten human health and have always been one of the refractory conditions facing humanity.Currently,gene therapy drugs such as siRNA,shRNA,antisense oligonucleotide,CRISPR/Cas9 system,plasmid DNA and miRNA have shown great potential in biomedical applications.To avoid the degradation of gene therapy drugs in the body and effectively deliver them to target tissues,cells and organelles,the development of excellent drug delivery vehicles is of utmost importance.Viral vectors are the most widely used delivery vehicles for gene therapy in vivo and in vitro due to their high transfection efficiency and stable transgene expression.With the development of nanotechnology,novel nanocarriers are gradually replacing viral vectors,emerging superior performance.This review mainly illuminates the current widely used gene therapy drugs,summarizes the viral vectors and non-viral vectors that deliver gene therapy drugs,and sums up the application of gene therapy to treat genetic diseases.Additionally,the challenges and opportunities of the field are discussed from the perspective of developing an effective nano-delivery system. 展开更多
关键词 gene therapy drugs Viral vectors Non-viral vectors genetic diseases Nano-delivery system
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Transduction of Fas gene or Bcl-2 antisense RNA sensitizes cultured drug resistant gastric cancer cells to chemotherapeutic drugs 被引量:24
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作者 XIAO Bing, SHI Yong Quan, ZHAO Yan Qiu, YOU Han, WANG Zuo You, LIU Xian Ling, YIN Fang, QIAO Tai Dong and FAN Dai Ming 《World Journal of Gastroenterology》 SCIE CAS CSCD 1998年第5期58-62,共5页
INTRODUCTIONChemotherapyisoneofthemajormethodsintumortreatment,butitoftendoesnotworkduetomultidrugresistance... INTRODUCTIONChemotherapyisoneofthemajormethodsintumortreatment,butitoftendoesnotworkduetomultidrugresistance(MDR).Recentstudi... 展开更多
关键词 stomach neoplasms FAS gene Bcl 2 gene ANTISENSE nucleic acid drug resistance multiple gene TRANSDUCTION apoptosis
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Clinical Study of Multi-drug Resistance Gene(MDR1) Expression in Primary Ovarian Cancer 被引量:1
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作者 王世宣 蔡桂茹 《Journal of Huazhong University of Science and Technology(Medical Sciences)》 SCIE CAS 1998年第1期58-60,共3页
This study was designed to measure the multi-drug resistance gene (MDR1) mRNA content and analyze clinical relationship between MDR1 expression and drug resistance in primary ovarian cancer. Reverse transcription PCR ... This study was designed to measure the multi-drug resistance gene (MDR1) mRNA content and analyze clinical relationship between MDR1 expression and drug resistance in primary ovarian cancer. Reverse transcription PCR (RT-PCR) was used to measure MDR1 mRNA content in biopsy sample of 31 primary ovarian cancers (experimental group) and 30 gynecological tumors (control group). The level of 95.2% (20/21) MDR1 expression was relatively low, and the detected rate of MDR1 expression was 67.7%(21/31) in experimental group,which was higher than that in control group (40.0%, P<0.05). The differences of MDR1 expression between the effective group and no effect group after combined chemotherapy was significant (P<0.05). No significant relationship was found between MDR1 expression and clinical stage or histological classification or grade of differentiation in experimental group. We are led to concluded that primary ovarian cancers have drug-resistance clones which might express MDR1 spontaneously and expression of MDR1 may be used as a prognostic and predictive indicator for clinical response of ovarian cancers to combined chemotherapy. 展开更多
关键词 OVARIAN neoplasma gene drug RESISTANCE CHEMOTHERAPY PCR
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Drug resistance gene expression and chemotherapy sensitivity detection in Chinese women with different molecular subtypes of breast cancer 被引量:1
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作者 Jing Zhao Hailian Zhang +5 位作者 Ting Lei Juntian Liu Shichao Zhang Nan Wu Bo Sun Meng Wang 《Cancer Biology & Medicine》 SCIE CAS CSCD 2020年第4期1014-1025,共12页
Objective:The aim of the study was to identify specific chemosensitivity drugs for various molecular subtypes of breast tumors in Chinese women,by detecting the expression of drug resistance genes and by using the dru... Objective:The aim of the study was to identify specific chemosensitivity drugs for various molecular subtypes of breast tumors in Chinese women,by detecting the expression of drug resistance genes and by using the drug sensitivity test on different molecular subtypes of breast cancers.Methods:The expression of drug resistance genes including Topo Ⅱ,GST-π,P-gp,LRP,and CD133 were detected with immunohistochemistry in a tissue microarray.Drug sensitivity tests included those for paclitaxel,epirubicin,carboplatin,vinorelbine,and fluorouracil and were conducted on primary cancer tissue cells and cell lines,including the T47 D,BT-474,and MDA-MB-231 cells and human breast cancer xenografts in nude mice.Results:The different drug resistant genes Topo Ⅱ,GST-π,P-gp,and LRP were differentially expressed among different molecular subtypes of breast cancers(P<0.05).Positive expression of CD133 was highest in basal-like breast cancer(P<0.05).Kaplan-Meier survival analysis showed that positive expressions of Topo Ⅱ and CD133 both correlated with shorter disease-free survival(DFS)(P<0.05)and overall survival(P<0.05),and positive expression of LRP correlated only with shorter DFS(P<0.05).BT-474 showed chemosensitivity to paclitaxel and epirubicin,while MDA-MB-231 showed chemosensitivities to paclitaxel,epirubicin,carboplatin,and fluorouracil(T/C≤50%).The basal-like and HER2+breast cancer primary cells showed chemosensitivities to paclitaxel and epirubicin with significant differences compared with luminal breast cancer primary cells(P<0.05).Conclusions:The differential expression of drug resistance genes and the differential chemosensitivities of drugs in different molecular subtype of breast cancers suggested that individual treatment should be given for each type of breast cancer. 展开更多
关键词 Breast cancer molecular subtype CD133 drug resistant gene CHEMOSENSITIVITY
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Anticancer Drug Resistance of HeLa Cells Transfected With Rat Glutathione S-transferase pi Gene 被引量:2
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作者 WEICAO YANMENG +3 位作者 QIANGWEI ZHAO-HUISHI LI-MEIJU FU-DEFANG 《Biomedical and Environmental Sciences》 SCIE CAS CSCD 2003年第2期157-162,共6页
To establish a cytologic expressing system of rat glutathione S-transferase pi (GST-pi) cDNA for detecting the resistance of HeLa cells to anticancer drugs. Methods The assessment was made with various anticancer drug... To establish a cytologic expressing system of rat glutathione S-transferase pi (GST-pi) cDNA for detecting the resistance of HeLa cells to anticancer drugs. Methods The assessment was made with various anticancer drugs (adriamycin, mitomycin, cisplatinum and vincristine) that showed different cytotoxicities in transfectant HeLa cells with pSV-GT containing rat GST-pi cDNA (HeLa/pSV-GT) or control pSV-neo (HeLa/pSV-neo). Expression levels of GST-pi mRNA in HeLa/pSV-GT and HeLa/pSV-neo were measured by in situ hybridization using Digoxin-labelled cDNA probe. Results HeLa/pSV-GT expressed significantly high degree of GST-pi mRNA, whereas both HeLa/pSV-neo and HeLa cells had very low expression. Cytotoxicities of HeLa/pSV-GT and HeLa/pSV-neo with 4 anticancer drugs were measured by MTT assay. Drug concentrations for yielding 50% inhibition (IC50) in HeLa/pSV-GT by adriamycin, mitomycin and cisplatinum were 70.13 靏/mL, 10.95 靏/mL and 16.52 靏/mL, respectively. In contrast, IC50 in HeLa/pSV-neo was 10.34 靏/mL, 7.48 靏/mL and 13.70 靏/mL, respectively. The cytotoxicities of vincristine on both HeLa/pSV-GT and HeLa/pSV-neo were not significantly different. Conclusions Our findings suggest that HeLa/pSV-GT containing rat GST-pi cDNA is resistant to some anticancer drugs due to overexpression of GST-pi. Also, HeLa/pSV-GT cell line could serve as a useful cytogenetic model for further research. 展开更多
关键词 还原型谷胱甘肽S转移酶 抗肿瘤药 HELA细胞 基因表达 肿瘤生成 作用机制
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Expression and Prognostic Significance of Multidrug Resistance Associated Protein (MRP) Gene in Non-small Cell Lung Cancer by in Site Hybridization 被引量:1
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作者 单根法 钟竑 +4 位作者 张辅贤 李国庆 隆桂麟 顾鹤定 戚晓敏 《Chinese Journal of Cancer Research》 SCIE CAS CSCD 2000年第3期63-66,共4页
Objective: To study on the effect of MRP gene overexpression on prognosis of patients with non-small lung cancer (NSCLC). Methods: Paraffin-embedded tissues from 47 cases of NSCLC who had undergone radical tumor resec... Objective: To study on the effect of MRP gene overexpression on prognosis of patients with non-small lung cancer (NSCLC). Methods: Paraffin-embedded tissues from 47 cases of NSCLC who had undergone radical tumor resection were examined for expression of MRP gene mRNA by in situ hybridization using labelled digoxigenin probes combined with immunohistochemistry. All the patients were retrospectively followed-up. Results: All of the 47 lung cancer specimens were found to have overexpression of MRP gene mRNA. It was significantly correlated with patients’ survival time, response to chemotherapy, recurrence or metastases after surgery, but was not correlated with histology, tumor size, node status, TNM stage, degree of differentiation, age and sex. Conclusion: Overexpression of MRP gene is a marker of prognostic significance in patients with NSCLC. 展开更多
关键词 LUNG NEOPLASMS multi-drug resistance MRP gene prognosis
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In vitro study of the influence of GST-π gene transfer on drug-resistance of human cord blood CD34^+ cells 被引量:1
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作者 YuChenghao YangXingsheng +1 位作者 CuiBaoxia JiangJie 《现代妇产科进展》 CSCD 2003年第3期238-240,共3页
Objective:To investigate the influence of GST-π gene transfer on drug-resistance of human cord blood CD34 + cells.Methods:CD34 + cells were purified from cord blood from normal full-term pregnancy.Gene transduction i... Objective:To investigate the influence of GST-π gene transfer on drug-resistance of human cord blood CD34 + cells.Methods:CD34 + cells were purified from cord blood from normal full-term pregnancy.Gene transduction into human cord blood CD34 + cells was carried out using GST-π gene containing retrovirus vector.The GST-π gene expression in transduced CD34 + cell was confirmed by RT-PCR.After confirmation of GST-π gene transfer,the transfected CD34 + cells were cultured by colony assay in the presence of carboplatin.Results:GST-π mRNA was detected in 30% of CFU-GM derived from GST-π gene transduced CD34 + cells.In vitro drug resistance test showed that the number of CFU-GM formed was significantly higher (2~3 fold) in GST-π gene transduced CD34 + cells than untransduced CD34 +cells.Conclusion:GST-π gene transfer can confer resistance to hematopoietic progenitors against carboplatin in vitro. 展开更多
关键词 CD34^细胞 GST-Π基因 耐药 RT-PCR 化疗 恶性肿瘤
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EXPRESSION AND REVERSION OF DRUG RESISTANCE-AND APOPTOSIS-RELATED GENES OF A DDP-RESISTANT LUNG ADENOCARCINOMA CELL LINE 被引量:1
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作者 王洁 张叙仪 蒋薇 《Chinese Journal of Cancer Research》 SCIE CAS CSCD 2000年第2期79-86,共8页
Objective: To investigate the co-expression of drug resistance- and apoptosis-related genes of cisplatin (CDDP)-selected lung adenocarcinoma cell line A549DDP for compared to the parental cell line A549, and reverse... Objective: To investigate the co-expression of drug resistance- and apoptosis-related genes of cisplatin (CDDP)-selected lung adenocarcinoma cell line A549DDP for compared to the parental cell line A549, and reverse of drug resistance by antisense s-oligodeoxynucleotides (S-ODNs) of differentially expressed genes. Methods: Sense and antisense S-ODN were transferred into A549DDP cells by lipofectin. The expression of drug resistance and apoptosis related genes was examined by RT-PCR, immunocytochemistry and flow cytometry, respectively. Apoptostic cells were identified by DNA electrophoresis and terminal deoxynucleotidyl transferase (TdT)-mediated biotin dUTP nick end- labeling(TUNEL). Drug resistance of tumor cells was detected by a cell viability (MTT) assay. Results: The expression of bc1-2 was positive and that of multidrug resistance-associated protein (MRP) at mRNA and protein level was increased in A549DDP compared to A549 cells. MDRI, c-myc and topoisomeras Ⅱ (TOPO Ⅱ) were similarly co-expressed in two cell lines. Both cell lines were negative for c-erbB-2 expression. In A549DDP cells, the expression of bc1-2 and MRP was sigllificantly inhibited by their respective antisense S-ODNs. Antisensc S-ODNs could also decrease significantly drug resistance of A549DDP cells to CDDP by promoting cell apoptosis. Conclusion: Both intrinsic and acquired drug resistance were involved in co-expression of multiple MDR-related genes in lung adenocarcinoma. Co- operation of hcf-2 and MRP genes appeared to play an important action to confer the resistance of A549DDP cells to CDDP. Their antisense S-ODNs are responsible for the decrease of drug resistance of this cell line by promoting apoptosis. 展开更多
关键词 Lung NEOPLASM A549 and A549DDP cell lines Apoptosis ANTISENSE oligoxynucleotidc drug resistance- gene
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Liposome-mediated Functional Expression of Multiple Drug Resistance Gene in Human Bone Marrow CD34^+ Cells
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作者 曹文静 邹萍 《Journal of Huazhong University of Science and Technology(Medical Sciences)》 SCIE CAS 2004年第3期214-215,235,共3页
Summary: The expression and functional activity of multiple drug resistance (MDR1) gene in human normal bone marrow CD34+ cells was observed. Human normal bone marrow CD34+ cells were enriched with magnetic cell sorti... Summary: The expression and functional activity of multiple drug resistance (MDR1) gene in human normal bone marrow CD34+ cells was observed. Human normal bone marrow CD34+ cells were enriched with magnetic cell sorting (MACS) system, and then liposome-mediated MDR1 gene was transferred into bone marrow CD34+ cells. Fluorescence-activated cell sorter was used to evaluate the expression and functional activity of P-glycoprotein (P-gp) encoded by MDR1 gene. It was found that the purity of bone marrow CD34+ cells was approximately (91±4.56) % and recovery rate was (72.3±2.36) % by MACS. The expression of P-gp in the transfected CD34+cells was obviously higher than that in non-transfected CD34+ cells. The amount of P-gp in non-transfected CD34+ cells was (11.2±2.2) %, but increased to (23.6±2.34) % 48 h after gene transfection (P<0.0l). The amount of P-gp was gradually decreased to the basic level one week later. The accumulation and extrusion assays showed that the overexpression of P-gp could efflux Rh-123 out of cells and there was low fluorescence within the transfected cells. The functional activity of P-gp could be inhibited by 10 μg/ml verapamil. It was suggested that the transient and highly effective expression and functional activity of P-gp could be obtained by liposome-mediated MRD1 transferring into human normal bone marrow CD34+ cells. 展开更多
关键词 脂肪体介导 功能表达 麻药抵抗基因 骨髓CD34+细胞 基因转染
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THE CLINICAL SIGNIFICANCE OF MULTIDRUG RESISTANCE GENE(mdr1) EXPRESSION IN ACUTE LEUKEMIA
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作者 董作仁 罗建民 +7 位作者 徐文新 王福旭 郭晓楠 张学军 姚尔固 徐世荣 任金海 丛斌 《Chinese Journal of Cancer Research》 SCIE CAS CSCD 1998年第1期61-64,共4页
THECLINICALSIGNIFICANCEOFMULTIDRUGRESISTANCEGENE(mdr1)EXPRESSIONINACUTELEUKEMIADongZuoren董作仁LuoJianmin罗建民XuW... THECLINICALSIGNIFICANCEOFMULTIDRUGRESISTANCEGENE(mdr1)EXPRESSIONINACUTELEUKEMIADongZuoren董作仁LuoJianmin罗建民XuWenxin徐文新WangFuxu... 展开更多
关键词 LEUKEMIA drug RESISTANCE gene EXPRESSION POLYMERASE chain reaction.
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Expression patterns and action analysis of genes associated with drug-induced liver diseases during rat liver regeneration 被引量:3
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作者 Qian-Ji Ning Shao-Wei Qin Cun-Shuan Xu 《World Journal of Gastroenterology》 SCIE CAS CSCD 2006年第43期6966-6972,共7页
AIM: To study the action of the genes associated with drug-induced liver diseases at the gene transcriptional level during liver regeneration (LR) in rats. METHODS: The genes associated with drug-induced liver disease... AIM: To study the action of the genes associated with drug-induced liver diseases at the gene transcriptional level during liver regeneration (LR) in rats. METHODS: The genes associated with drug-induced liver diseases were obtained by collecting the data from databases and literature, and the gene expression changes in the regenerating liver were checked by the Rat Genome 230 2.0 array. RESULTS: The initial and total expression numbers of genes occurring in phases of 0.5-4 h after partial hepatectomy (PH), 4-6 h after PH (G0/G1 transition), 6-66 h after PH (cell proliferation), 66-168 h after PH (cell differentiation and structure-function reconstruction) were 21, 3, 9, 2 and 21, 9, 19, 18, respectively. It is illustrated that the associated genes were mainly triggered at the initial stage of LR and worked at different phases. According to their expression similarity, these genes were classified into 5 types: only up- regulated (12 genes), predominantly up-regulated (4 genes), only down-regulated (11 genes), predominantly down-regulated (3 genes), and approximately up-/ down-regulated (2 genes). The total times of their up- and down-expression were 130 and 79, respectively, demonstrating that expression of most of the genes was increased during LR, while a few decreased. The cell physiological and biochemical activities during LR were staggered according to the time relevance and were diverse and complicated in gene expression patterns. CONCLUSION: Drug metabolic capacity in regenerating liver was enhanced. Thirty-two genes play important roles during liver regeneration in rats. 展开更多
关键词 基因表达 肝再生 药物治疗 临床
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温州市食源性小肠结肠炎耶尔森氏菌毒力基因分布及遗传多样性研究
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作者 谢爱蓉 李毅 +4 位作者 楼辉煌 谢中必 章乐怡 胡玉琴 吴跃进 《中国人兽共患病学报》 CAS CSCD 北大核心 2024年第1期40-45,共6页
目的了解温州市食源性小肠结肠炎耶尔森氏菌的毒力基因分布及遗传多样性特征。方法对温州市食品及食源性疾病腹泻病人中分离得到的小肠结肠炎耶尔森氏菌进行生物型、血清型和耐药性分析,并基于全基因组测序进行菌株的毒力基因、多位点... 目的了解温州市食源性小肠结肠炎耶尔森氏菌的毒力基因分布及遗传多样性特征。方法对温州市食品及食源性疾病腹泻病人中分离得到的小肠结肠炎耶尔森氏菌进行生物型、血清型和耐药性分析,并基于全基因组测序进行菌株的毒力基因、多位点序列分型(MLST)、核心基因组多位点序列分型(cgMLST)分析。结果71株食源性小肠结肠炎耶尔森氏菌,94.4%(67/71)的分离株为生物1A型,生物血清型以1A/O∶5占比最高(29.6%,21/71)。分离株对14种抗菌药物的敏感率达到95.8%以上。共获得了16种126个毒力基因,其中2株菌株携带pYV质粒和染色体相关毒力基因。分离株以ST3型最常见(31.6%,12/38),cgMLST分析显示除具有相同ST型的菌株外,菌株无明显基因型簇形成。结论温州市食源性小肠结肠炎耶尔森氏菌存在致病性菌株,菌株呈高度遗传多态性,需加强此类菌株的监测,以预防其引起食源性疾病的暴发。 展开更多
关键词 小肠结肠炎耶尔森氏菌 血清型 毒力基因 耐药性 全基因组测序
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2018—2019年辽宁省副溶血性弧菌临床和食品分离株分子流行病学分析
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作者 李雪 孙婷婷 +2 位作者 魏彤竹 王伟杰 刁文丽 《中国人兽共患病学报》 CAS CSCD 北大核心 2024年第3期283-288,共6页
目的了解辽宁省副溶血性弧菌(Vibrio Parahaemolyticus,VP)的流行特征,为辽宁省食源性疾病的防控和应急事件的处置提供可靠的技术支持。方法对2018—2019年辽宁省301株临床和食品VP分离株进行血清学分型、脉冲场凝胶电泳(PFGE)分型、毒... 目的了解辽宁省副溶血性弧菌(Vibrio Parahaemolyticus,VP)的流行特征,为辽宁省食源性疾病的防控和应急事件的处置提供可靠的技术支持。方法对2018—2019年辽宁省301株临床和食品VP分离株进行血清学分型、脉冲场凝胶电泳(PFGE)分型、毒力基因(tlh、tdh、trh)鉴定及耐药试验。结果除了59株VP分离株不能进行血清学分型外,其他分离株共分为17个血清型,血清型以O3群、O1群和O2群为主。血清型O3与O1群分离株相似度高。PFGE主要分成11组优势带型,A-K组,其中A-G组为主要流行致病分子型。临床分离株毒力基因携带情况主要为tlh+、tdh+、trh-,食品分离株毒力基因携带情况主要为tlh+、tdh-、trh-。VP分离株主要对β-内酰胺类(CFZ和AMP)、大环内酯类抗菌药物(ERY)耐药。即食食品分离株耐药率远高于非即食食品分离株。结论由于即食食品的特殊性和来自于国外的特性,这些菌株具有引起食源性疾病暴发流行的高风险,应高度重视。部分食品分离株也具有引起食源性疾病的风险,应给予关注。相关部门除了应加强对国内海产品及其抗菌药物使用管控外,更应加强对进口海产品的监管。 展开更多
关键词 副溶血性弧菌 毒力基因 血清学分型 耐药
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北京某医院感染性腹泻患者致泻性大肠埃希菌毒力和耐药特征分析
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作者 马思宇 马立艳 +3 位作者 罗虎 周妍妍 王雅轩 苏建荣 《临床检验杂志》 CAS 2024年第1期43-47,共5页
目的明确本院感染性腹泻患者致泻性大肠埃希菌毒力和耐药特征。方法采用VITEK MS微生物质谱检测系统初步鉴定,多重实时荧光PCR检测毒力基因,对本院感染性腹泻患者临床分离的致泻性大肠埃希菌(Diarrheagenic Escherichia coli,DEC)进行5... 目的明确本院感染性腹泻患者致泻性大肠埃希菌毒力和耐药特征。方法采用VITEK MS微生物质谱检测系统初步鉴定,多重实时荧光PCR检测毒力基因,对本院感染性腹泻患者临床分离的致泻性大肠埃希菌(Diarrheagenic Escherichia coli,DEC)进行5种型别鉴定。微量肉汤稀释法和E-test法药敏试验检测DEC菌株的耐药表型特征。二代测序及生物信息学分析其耐药分子特征。采用Fisher确切概率法进行统计学分析。结果本院DEC检出率为11.9%,其中EAEC占比37.5%,非典型EPEC占比34.38%,ETEC占比25.0%,EIEC占比3.12%,未检出EHEC菌株。32株DEC对氨苄西林、四环素、甲氨苄啶/磺胺异恶唑耐药率最高,分别为53.12%、43.75%和37.5%。ESBLs(+)株占比18.75%,其多重耐药菌株检出率为83.83%,显著高于ESBLs(-)菌株,差异有统计学意义(P=0.042)。32株DEC共有25个ST型,优势基因型为ST10共4株(12.5%),ST28、ST31和ST3153各2株(各占比6.25%),其他21个型别各1株菌(各占比3.13%)。EAEC中检出1株携带bla_(NDM-1)的耐碳青霉烯类菌株。结论我院感染性腹泻患者的DEC以aggR、pic、astA、eae 4种毒力基因较为常见,以EAEC、EPEC为主要型别,基因型别呈高度多态性,检出多重耐药菌株。 展开更多
关键词 致泻性大肠埃希菌 毒力基因 耐药基因
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湖南省部分地区禽致病性大肠杆菌分离鉴定及生物特性分析
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作者 刘伯承 燕海峰 +4 位作者 罗阳 王慧 杨俊 陈一峰 周望平 《中国动物检疫》 CAS 2024年第3期28-37,共10页
为探究湖南省禽致病性大肠杆菌的生物特性,从部分地区规模肉鸡场采集122份病死鸡临床肝脏样品,采用传统细菌培养方法进行分离培养和初步鉴定,利用大肠杆菌持家基因phoA进行PCR扩增,对纯化鉴定后的分离株进行血清型鉴定、种系发育群检测... 为探究湖南省禽致病性大肠杆菌的生物特性,从部分地区规模肉鸡场采集122份病死鸡临床肝脏样品,采用传统细菌培养方法进行分离培养和初步鉴定,利用大肠杆菌持家基因phoA进行PCR扩增,对纯化鉴定后的分离株进行血清型鉴定、种系发育群检测、多位点序列分型(MLST)、毒力基因检测、动物致病性试验及耐药性分析。结果显示:共分离出83株禽致病性大肠杆菌,鉴定出15种血清型,其中O2、O1、O78、O14为优势血清型,种系发育群分为B2、B1、A和D群,高致病性株主要来自B2、B1群;MLST分析共得到12种ST型,其中ST95、ST481、ST350为主要ST型;分离株irp2、fyuA、iutA、fimC毒力基因携带率为100%,其中38.55%菌株同时携带irp2、afa、fyuA、iutA、hlyF、iss、papC、fimC毒力基因;雏鸡致病性试验检出高致病性菌株46株(55.42%)、中等致病性菌株31株(37.35%)、低等致病性菌株6株(7.23%);分离株对10种常用抗生素有不同程度耐药,其中对四环素和新生霉素耐药最严重,耐药率分别为96.38%和92.77%。结果表明,湖南省禽致病性大肠杆菌遗传关系整体复杂多态,且致病性较高,耐药广泛,提示该地区应加强对禽大肠杆菌病的防控。 展开更多
关键词 禽致病性大肠杆菌 血清型 毒力基因 MLST 耐药性
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拟杆菌属临床分离株耐药性及脆弱拟杆菌肠毒素bft基因分型特征分析
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作者 王艳艳 王俊瑞 +2 位作者 郑文琪 兰海霞 郭素芳 《检验医学》 CAS 2024年第1期47-52,共6页
目的 分析拟杆菌属临床分布及其耐药特征,并探讨脆弱拟杆菌肠毒素bft基因分型特征。方法收集2016年6月—2019年6月内蒙古医科大学附属医院分离自临床样本的拟杆菌属147株,对其进行菌种鉴定和体外药物敏感性试验。采用聚合酶链反应(PCR)... 目的 分析拟杆菌属临床分布及其耐药特征,并探讨脆弱拟杆菌肠毒素bft基因分型特征。方法收集2016年6月—2019年6月内蒙古医科大学附属医院分离自临床样本的拟杆菌属147株,对其进行菌种鉴定和体外药物敏感性试验。采用聚合酶链反应(PCR)检测脆弱拟杆菌肠毒素bft基因。结果 147株拟杆菌属细菌中,脆弱拟杆菌100株(68.0%)、多形拟杆菌25株(17.0%)、卵形拟杆菌9株(6.1%)。体外药物敏感性试验结果显示,拟杆菌属细菌对克林霉素的耐药率最高(78.9%);脆弱拟杆菌对克林霉素的耐药率为85.0%,高于其他拟杆菌(66.0%)。拟杆菌属细菌对亚胺培南和甲硝唑的耐药率分别为13.6%和4.7%,对其他抗菌药物呈不同程度耐药;脆弱拟杆菌对大部分抗菌药物的耐药率均高于其他类型拟杆菌。脆弱拟杆菌bft基因阳性率为38.0%,以bft-1亚型为主要基因型(28.0%),其次为bft-2亚型(10.0%),未检出bft-3亚型。分离自非腹部临床样本的脆弱拟杆菌bft基因阳性率(55.6%)高于分离自腹部临床样本的脆弱拟杆菌(36.3%)。结论 拟杆菌属临床分离株中脆弱拟杆菌分离率最高,其对临床常用抗菌药物均有一定的耐药性。分离自非腹部临床样本的脆弱拟杆菌产肠毒素的菌株流行率更高,应引起临床重视。 展开更多
关键词 拟杆菌属 脆弱拟杆菌 耐药性 bft基因
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施用猪粪水对青脆李果实品质和产量、土壤中微生物群落与抗生素耐药基因的影响
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作者 雷云峰 何志平 +5 位作者 龚建军 肖璐 涂腾 曾凯 付敏 曾祥忠 《西南农业学报》 CSCD 北大核心 2024年第1期189-198,共10页
【目的】为了解猪粪浇灌对青脆李果实品质和产量、土壤性质、土壤中微生物群落与抗生素耐药基因的影响。【方法】以5年生青脆李作试材,设置猪粪水按不同比例(100%、75%、50%、25%、0%)替代化肥田间试验,通过单果重、单果横径等外观指标... 【目的】为了解猪粪浇灌对青脆李果实品质和产量、土壤性质、土壤中微生物群落与抗生素耐药基因的影响。【方法】以5年生青脆李作试材,设置猪粪水按不同比例(100%、75%、50%、25%、0%)替代化肥田间试验,通过单果重、单果横径等外观指标,李子每667 m^(2)产量,维生素C、可溶性固形物等果实品质指标测定,分析不同施肥处理对青脆李果实品质和产量的影响;基于土壤中pH、有机质含量等理化指标测定,分析不同施肥处理对李树土壤理化性质的影响;基于Illumina Hiseq 4000测序平台对李树土壤宏基因组测序,分析猪粪水浇灌对土壤中微生物群落变化、抗生素耐药基因分布的影响。【结果】不同施肥处理青脆李果实品质和产量研究结果表明,“75%猪粪+25%化肥”“50%猪粪+50%化肥”处理青脆李果实外观品质均较优,“50%猪粪+50%化肥”处理青脆李每667 m^(2)产量最高。不同施肥处理对李树土壤理化性质的研究结果显示,各处理间有机质、碱解氮等理化指标均不存在显著性差异(P>0.05)。宏基因组测序分析结果显示,“100%猪粪”组中的优势菌群为放线菌,而其余4组的优势菌群均为假单胞菌门;共鉴定39个抗生素本体(ARO),随着猪粪水用量的减少,土壤样品中检出ARO种类、ARO相对丰度和均呈依次递减趋势。【结论】本研究条件下,猪粪水部分替代化肥为青脆李树施肥是可行的,且以“50%猪粪+50%化肥”进行施肥最佳。 展开更多
关键词 青脆李 猪粪 土壤 菌群 宏基因组 抗生素 耐药基因
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主穹窿蛋白基因在高原地区癫痫儿童中的多态性分析
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作者 冶秀兰 李明 +6 位作者 王兆建 赵海燕 马建宁 刘婧 郑怡 李顺清 张玉 《吉林医学》 CAS 2024年第5期1034-1037,共4页
目的:探讨分析主穹窿蛋白(MVP)基因三个SNP位点rs4788187、rs3815824、rs3815823多态性与高原地区儿童癫痫的相关性。方法:选取2017年1月~2020年1月青海大学附属医院收治的80例癫痫患儿为研究对象,根据耐药性不同分为耐药组与非耐药组... 目的:探讨分析主穹窿蛋白(MVP)基因三个SNP位点rs4788187、rs3815824、rs3815823多态性与高原地区儿童癫痫的相关性。方法:选取2017年1月~2020年1月青海大学附属医院收治的80例癫痫患儿为研究对象,根据耐药性不同分为耐药组与非耐药组各40例,利用PCR-RELP方法检测rs4788187、rs3815824、rs3815823三个SNP位点的多态性分布,并进行统计分析。结果:两组年龄、性别、民族以及癫痫发作类型比较差异无统计学意义(P>0.05)。癫痫患儿rs4788187、rs3815824的等位基因C频率明显低于T,而rs3815823的等位基因T频率明显低于C;两组患儿SNP位点rs4788187、rs3815824、rs3815823的基因型频率和等位基因频率比较差异均无统计学意义(P>0.05)。结论:MVP基因的SNP位点(rs4788187、rs3815824、rs3815823)与癫痫耐药性不相关。 展开更多
关键词 儿童 癫痫 主穹窿蛋白 耐药 基因多态性
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多重耐药弗劳地枸橼酸杆菌的基因组及耐药机制分析
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作者 黄蓉 段小女 芮勇宇 《国际检验医学杂志》 CAS 2024年第7期785-789,共5页
目的对多重耐药弗劳地枸橼酸杆菌进行全基因组分析,为研究其耐药机制提供依据。方法采用含美罗培南的MH培养基对粪便标本进行初筛,经BD Phoenix100全自动微生物鉴定仪进行菌种鉴定及药敏试验,提取耐药菌总DNA进行二代测序和细菌多位点... 目的对多重耐药弗劳地枸橼酸杆菌进行全基因组分析,为研究其耐药机制提供依据。方法采用含美罗培南的MH培养基对粪便标本进行初筛,经BD Phoenix100全自动微生物鉴定仪进行菌种鉴定及药敏试验,提取耐药菌总DNA进行二代测序和细菌多位点序列分型(MLST),经质粒全基因组序列分析检测质粒的组分和功能,并与参考质粒进行比较,通过质粒接合转移实验分析耐药质粒传递情况。结果413份粪便标本中分离出1株多重耐药弗劳地枸橼酸杆菌,该菌株对头孢菌素类、碳青霉烯类抗菌药物、复方磺胺甲噁唑的耐药率均为100%,对氨曲南、环丙沙星、左氧氟沙星、庆大霉素等抗菌药物呈现不同程度耐药;MLST分型的耐药弗劳地枸橼酸杆菌为ST22型,该菌株共含有5412个基因,含耐药基因367个,包括碳青霉烯酶类耐药基因、AmpC酶基因、大环内酯类耐药基因、氨基糖苷类耐药基、喹诺酮类耐药基因等。质粒的全基因组分析结果显示,该质粒含碳青霉烯类耐药基因blaNDM-1、blaSHV12,该质粒与泄殖腔肠杆菌菌株ECN49质粒和肺炎克雷伯菌菌株质粒pA575-NDM有极高的同源性;质粒接合实验显示,blaNDM-1和blaSHV-12可以通过质粒转移。结论弗劳地枸橼酸杆菌耐药的主要原因之一是含有blaNDM-1、blaSHV-12等耐药基因,并且该耐药性能通过质粒进行水平转移。 展开更多
关键词 弗劳地枸橼酸杆菌 全基因组分析 耐药基因 质粒
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2019—2021年皖南地区地方品种鸡源沙门菌的血清型鉴定、毒力基因和耐药性检测
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作者 吴萍萍 贺绍君 《中国兽医杂志》 CAS 北大核心 2024年第3期40-47,共8页
为了分析皖南地区地方品种鸡(淮南麻黄鸡、淮北麻鸡、霍邱鸡和五华鸡)来源沙门菌的分布、毒力基因和耐药性情况,本试验于2019—2021年采集该地区地方品种鸡养殖场疑似沙门菌感染病死鸡的肝脏组织、肛拭子和死胚等病料组织456份,采用细... 为了分析皖南地区地方品种鸡(淮南麻黄鸡、淮北麻鸡、霍邱鸡和五华鸡)来源沙门菌的分布、毒力基因和耐药性情况,本试验于2019—2021年采集该地区地方品种鸡养殖场疑似沙门菌感染病死鸡的肝脏组织、肛拭子和死胚等病料组织456份,采用细菌分离培养、形态学观察、生化试验和特异性invA基因的PCR检测进行沙门菌鉴定,采用Kauffmann-White法、PCR法和K-B纸片法分别检测分离菌株的血清型、毒力基因、耐药基因和耐药性。结果显示,共分离得到127株沙门菌,分属于13种血清型,其中以鸡白痢沙门菌、禽伤寒沙门菌和肠炎沙门菌为流行的优势血清型,分别占分离菌株的26.0%、20.5%和17.3%;127株沙门菌中携带的4种毒力基因(spvA、spvB、spvC和spvR)检出率介于7.1%~74.8%,携带的4种毒力岛基因(SPI-1 SPI-2、SPI-3和SPI-5)检出率介于38.6%~81.1%;127株沙门菌对阿莫西林、氨苄西林和磺胺二甲氧嘧啶等8种药物的耐药率介于56.7%~99.2%,以耐10(12.6%)、9(15.0%)和8(20.5%)种药物为主,耐药基因sul 1、sul 2、sul 3、tet(A)、tet(M)和tet(R)检出率介于32.3%~89.8%;经分析,地方品种鸡流行优势血清型与毒力基因和耐药基因均有一定的相关性。结果表明,从安徽省皖南地区4种地方品种鸡中分离的沙门菌具有多种血清型、携带多种毒力基因、耐药性严重,且携带多种耐药基因。 展开更多
关键词 皖南地区 地方品种鸡 沙门菌 血清型 耐药性 耐药基因
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