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PRRSV阳性猪源肺炎克雷伯菌的分离鉴定及特性分析

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中国兽医科学 2024年第11期54卷 1491-1501页

作者：常辉王玉淼李静李洋荆扬郑紫方裴秀秀陈劭肖书奇郑海学齐萌塔里木大学动物科学与技术学院新疆阿拉尔843300 中国农业科学院兰州兽医研究所兰州大学动物医学与生物安全学院动物疫病防控全国重点实验室甘肃兰州730046 西北农林科技大学动物医学院陕西杨凌712100

本试验旨在研究临床感染猪繁殖与呼吸综合征病毒(porcine reproductive and respiratory syndrome virus,PRRSV)后继发感染的细菌病原及其生物学特性,为实现PRRSV继发细菌感染的有效防控提供研究基础。无菌采集感染PRRSV死亡仔猪的胸腔... 详细信息

本试验旨在研究临床感染猪繁殖与呼吸综合征病毒(porcine reproductive and respiratory syndrome virus,PRRSV)后继发感染的细菌病原及其生物学特性,为实现PRRSV继发细菌感染的有效防控提供研究基础。无菌采集感染PRRSV死亡仔猪的胸腔积液进行细菌的分离与纯化,对分离菌进行形态学鉴定、分子生物学鉴定、药敏试验检测、耐药基因和毒力基因检测,进一步通过动物试验评估分离菌的致病性。结果发现,该菌为革兰阴性短杆菌,葡萄糖、尿素、蛋白胨水、枸橼酸盐、赖氨酸生化试验呈阳性,初步鉴定为克雷伯菌属细菌;细菌通用引物PCR扩增产物测序结果显示,该菌与巴基斯坦肉鸡源肺炎克雷伯菌的同源性为99.77%,进一步鉴定出分离菌株为肺炎克雷伯菌。药敏试验结果显示,该株肺炎克雷伯菌对四环素、红霉素、阿奇霉素、青霉素、氨苄西林、林可霉素6种药物有较强的耐药性,对链霉素、卡那霉素不敏感;该菌携带3种耐药基因和6种毒力基因,对BALB/c小鼠具有较强的致病性,7.0×10^(8)CFU/m L攻毒时死亡率为100%。本研究自PRRSV感染死亡仔猪体内分离出1株肺炎克雷伯菌,该菌具有多重耐药性和较强的致病性;肺炎克雷伯菌是PRRSV感染的重要继发致病菌,对PRRSV阳性猪继发细菌感染的防治具有重要指导意义。

关键词：猪繁殖与呼吸综合征病毒肺炎克雷伯菌毒力耐药性致病性

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胃肠超声评价功能性消化不良患者胃动力

莆田学院学报 2021年第5期28卷 36-40页

作者：曾宸陈薇陈海兰郑子芳林群莆田学院附属医院超声科福建莆田351100 莆田学院附属医院微创外科福建莆田351100 莆田学院附属医院核医学科福建莆田351100

探讨胃肠超声评价功能性消化不良(FD)患者胃动力价值。选取149例FD患者,包括上腹痛综合征(EPS)72例(EPS组)与餐后不适综合征(PDS)77例(PDS组),并选取健康体检者75例(对照组),比较各组胃肠超声检查参数。结果表明,应用胃肠超声能准确、... 详细信息

探讨胃肠超声评价功能性消化不良(FD)患者胃动力价值。选取149例FD患者,包括上腹痛综合征(EPS)72例(EPS组)与餐后不适综合征(PDS)77例(PDS组),并选取健康体检者75例(对照组),比较各组胃肠超声检查参数。结果表明,应用胃肠超声能准确、直观评价FD患者胃动力状态。

关键词：助显剂胃肠超声功能性消化不良胃动力 ROC曲线

Variation of iron species in sliding mud

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Chinese Science Bulletin 2002年第23期47卷 2018-2024页

作者： Guodong zheng Sheng Xu Yuhua Lang zifang Meng Chinese Acad Sci State Key Lab Gas GeochemLanzhou Inst GeolLanzhou 730000China

Results of Mossbauer spectroscopy and ICP-AES measurement revealed a special distribution pattern of iron species in four crossing profiles of the Tertiary Formation Type of Landslides in Japan. In comparison with host rocks, the sliding mud in the landslide slip zone contains the higher ferrous iron content and/or trace pyrite, which indicates that the forming environment was relatively reducing. The total iron content also increased with an increase in ferric iron and pyrite, in contrast, decreased with an increase in ferrous iron content. The deep-gray and black mud layers formed and accumulated in various volumes within landslide slip zones are treated as the basic criteria for the slipping activity, because the soft mud saturated with groundwater is easy to create a plastic deformation.

关键词： sliding mud Mossbauer spectroscopy iron speciation total iron slip zone redox condition

Mechanism of arterial remodeling in chronic allograft vasculopathy

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Frontiers of Medicine 2011年第3期5卷 248-253页

作者： Qichang zheng Shanglong Liu zifang Song Department of Hepatobiliary Surgery Union HospitalTongji Medical CollegeHuazhong University of Science and TechnologyWuhan 430022China

Chronic allograft vasculopathy(CAV)remains a major obstacle for long-term survival of grafts even though therapeutic strategies have improved considerably in recent *** is characterized by concentric and diffuse neointimal formation,medial apoptosis,infiltration of lymphocyte or inflammatory cells,and deposition of extracellular matrix both in arteries and *** studies have shown that stem cells derived from the recipient contribute to neointimal formation under the regulation of chemokines and *** remodeling in allografts eventually causes ischemic graft *** pathogenesis is multi-factorial with both immunologic and non-immunological factors being *** immunological factors have been discussed extensively in other *** review focuses mainly on the arterial remodeling that occurs in 3 layers of vessel walls including intimal injury,accumulation of smooth muscle-like cells in the neointimal,medial smooth muscle cell apoptosis,adventitial fibrosis,and deposition of extracellular matrix.

关键词： transplantation chronic rejection neointimal immunology arterial remodeling allograft vasculopathy

三种甲状腺术式创伤差异的前瞻性随机对照研究

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微创医学 2019年第4期14卷 435-439,445页

作者：简陈兴吴黎敏郑子芳方剑英刘伟涂海健莆田学院附属医院微创外科福建省莆田市351100 莆田学院附属医院药剂科福建省莆田市351100 莆田学院附属医院检验科福建省莆田市351100

目的通过前瞻性随机对照研究,比较三种不同甲状腺术式的创伤影响。方法150例患者按随机分配原则分为三组,每组50例,COT组采用常规开放甲状腺手术、EAT组采用腔镜辅助手术、TET组采用全腔镜手术。比较三组手术时间、术中出血量、不同时... 详细信息

目的通过前瞻性随机对照研究,比较三种不同甲状腺术式的创伤影响。方法150例患者按随机分配原则分为三组,每组50例,COT组采用常规开放甲状腺手术、EAT组采用腔镜辅助手术、TET组采用全腔镜手术。比较三组手术时间、术中出血量、不同时段外周血C反应蛋白(CRP)的水平、术后第1天引流量、术后拔管时间、术后第1天疼痛视觉模拟评分、切口美容满意度、术后3个月吞咽不适感等方面的差异。结果(1)所有手术均完成患侧腺叶切除(106例)或部分切除(44例)。EAT组、COT组中,未出现中转开放手术的病例。三组均无气管、甲状旁腺损伤以及不可控的术中大出血等并发症发生。术中冰冻切片病理结果均为良性。(2)三组手术时间、术中出血量、术后第1天引流量及拔管时间比较,差异均有统计学意义(P<0.05)。TET组的手术时间最长,COT组最短;TET组、EAT组的术中出血量明显少于COT组;EAT组的术后第1天引流量最少,TET组最多;EAT组拔管时间明显短于TET组、COT组(均P<0.05)。(3)三组患者术后6h、24h、72h的CRP水平比较,差异均有统计学意义(P<0.05)。术后6h,TET组CRP明显高于EAT组、COT组;术后24hEAT组最低;术后72h的CRP以EAT组最低、TET组最高(均P<0.05)。(4)三组患者术后第1天疼痛视觉模拟评分、美容满意度、术后3个月吞咽不适感比较,差异均有统计学意义(P<0.05)。EAT组术后第1天疼痛感明显低于TET组、COT组;TET组美容满意度最高,COT组最低;EAT组术后3个月吞咽不适感最轻,TET组最重(均P<0.05)。结论三种手术方式均为安全手术方式,就总体创伤性影响而言,腔镜辅助甲状腺手术应该是创伤最小的手术方式。临床实践中应根据患者美容需求和病情决定手术方式。

关键词：腔镜甲状腺手术入路创伤随机对照

Differences in the primary culture, purification and biological characteristics between endothelial cells and smooth muscle cells from rat aorta

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Journal of Nanjing Medical University 2009年第4期23卷 241-246页

作者： Shaobo Hu zifang Song Qichang zheng Jun Nie Department of Hepatobiliary Surgery Union Hospital Huazhong University of Science and Technology Wuhan 430022 China Department of Chest Surgery Union Hospital Huazhong University of Science and Technology Wuhan 430022 China

Objective： To investigate the differences of primary culture, purification and biological characteristics between endothelial cells and smooth muscle cells from rat aorta. Methods： Endothelial cells were obtained using the vascular ring adherence, collagenase digestion method and an improved vascular ring adherence method, while smooth muscle cells were separated from tissue sections of rat aorta. Clones of endothelial cells were selected by limiting dilution assay. Both cell types were identified using specific cell immunofluorescent markers, and phase contrast microscopy was used to observe the morphological disparity between endothelial cells and smooth muscle cells at the single cell and colony level. Cell proliferation was determined by the cell counting kit-8. Differences between endothelial cells and smooth muscle cells were evaluated in trypsin digestion time, attachment time and recovery after cryopreservation. Results： Endothelial cells were obtained by all three methods. The improved vascular ring method provided the most reproducible results. Cells were in good condition, and of high purity. Smooth muscle cells were cultured successfully by the tissue fragment culture method. Clonal expansion of single endothelial cells was attained. The two cell types expressed their respective specific markers, and the rate of proliferation of smooth muscle cells exceeded that of endothelial cells. Endothelial cells were more sensitive to trypsin digestion than smooth muscle cells. In addition, they had a shorter adherence time and better recovery following cryopreservation than smooth muscle cells. Conclusion： The improved vascular ring method was optimal for yielding endothelial cells. Limiting dilution is a novel and valid method for purifying primary endothelial cells from rat aorta. Primary rat endothelial cell and vascular smooth muscle cell cultures exhibited different morphological characteristics, proliferation rate, adherence time, susceptibility to trypsin d

关键词： endothelial cell smooth muscle cell smooth muscle like-cell cell culture

Selenium nanoparticles in aquaculture:Unique advantages in the production of Se-enriched grass carp(Ctenopharyngodon idella)

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Animal Nutrition 2024年第1期16卷 189-201页

作者： Chao Zhu zifang Wu Qimin Liu Xiaolin Wang Lijuan zheng Shuyang He Fangxia Yang Hong Ji Wuzi Dong College of Animal Science and Technology Northwest A&F UniversityYanglingShaanxi712100China College of Forestry Northwest A&F UniversityYanglingShaanxi712100China Biology Research Centre of Qin Mountains Wildlife Northwest A&F UniversityYanglingShaanxi712100China

The production of selenium-enriched fish can contribute to alleviating selenium deficiency in human ***,it is still unclear which selenium source,as an additive,can efficiently and costeffectively produce high-quality selenium-enriched *** study evaluated the effects of selenium nanoparticles(SeNP),selenite,and selenomethionine(SeMet)on the growth,antioxidant capacity,selenium content,selenium speciation,and meat quality of grass *** diets were prepared,including a basal diet(BD)and three concentrations(0.1,0.3,and 0.9 mg/kg)of SeNP,selenite,and SeMet.A total of600 fish(250.79±1.57 g)were randomly assigned to 30 tanks(3 tanks/group).Fish were fed the experimental diet three times daily for 60 *** this study,SeNP most significantly promoted the growth and antioxidant capacity of grass carp,with 0.3 mg/kg SeNP identified as the optimal additive ***,SeNP demonstrated equally excellent bioavailability as SeMet and significantly increased the content of SeMet in grass carp(Ctenopharyngodon idella)***,compared to SeMet and selenite,dietary SeNP could more significantly enhance the content of selenocysteine(SeCys2)and methylselenocysteine(MeSeCys)in grass carp muscle *** addition,we have demonstrated that SeCys2and MeSeCys promote apoptosis of cancer cells(HeLa)through the mitochondrial apoptotic pathway(involving Bax and Bcl-2).Furthermore,as an additive,0.3 mg/kg SeNP significantly improved the flesh quality of grass carp by reducing crude fat and heavy metal content,as well as increasing the levels of eicosapentaenoic acid(EPA)and docosahexaenoic acid(DHA)and the ratio of n-3/n-6 polyunsaturated fatty acid(PUFA).In summary,SeNP is the most suitable additive for producing selenium-enriched fish.

关键词： Selenium nanoparticle Speciation analysis Grass carp Selenium additive

Construction of eukaryotic expression vector of humanarresten gene and its secreted expression in HEK 293 cells

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Frontiers of Medicine 2009年第3期3卷 297-302页

作者： Wei LI zifang SONG Qichang zheng Jun XIONG Dan SHANG Siming GUAN Xiaogang SHU Department of Gerontology Union HospitalTongji Medical CollegeHuazhong University of Science and TechnologyWuhan 430022China Department of General Surgery Union HospitalTongji Medical CollegeHuazhong University of Science and TechnologyWuhan 430022China

The eukaryotic expression vector of human arresten gene was constructed and its secretive expression in human embryonic kidney(HEK 293)cells was *** arresten gene was ampliﬁed from recombinant plasmid pGEM-Arr by polymerase chain reaction(PCR),and then digested with restriction endonucleases BamH I and EcoR *** target fragment was inserted into the BamH I and EcoR I restriction sites of eukaryotic expression vector pSecTag2 to construct ***-tion analysis and DNA sequencing indicated that the arresten gene was successfully inserted into *** recombinant plasmid was subsequently transfected into HEK 293 cells with LipofectAMINETM2000 Reagent,and the expression of the target gene was *** transcription PCR(RT-PCR)revealed that the mRNA of the target gene was transcribed in the transfected HEK 293 *** blot analysis veriﬁed that the recombinant protein in supernatants was *** supernatants of transfected cells were prepared,and 3-(4,5)-dimethylthiazol(-2-y1)-3,5-di-phenyltetrazoliumbromide(MTT)assay was carried out to assess their effects on the proliferation of human umbilical vein endothelial cells,which showed that the recombinant protein could signiﬁcantly suppress the proliferation of human umbilical vein endothelial cells in *** results provided a solid foundation to explore the usage of arresten in tumor anti-angiogenic gene therapy.

关键词： angiogenesis inhibitor arresten eukaryotic expression HEK 293 cells endothelial cells

The effect of human papilloma virus type 16 E7 protein expression on growth of RMA cells in vitro and in vivo

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The Chinese-German Journal of Clinical Oncology 2010年第2期9卷 73-78页

作者： Shaobo Hu Qichang zheng zifang Song Xiedan Wang Department of Hepatobiliary Surgery Union Hospital Huazhong University of Science and Technology Wuhan 430022 China Department of General Surgery The Affiliated Hospital of Wuhan University of Science and Technology Wuhan 430064 China

Objective： The aim of this study was to study the effect of human papilloma virus （HPV） type 16 E7 protein ex- pression on growth of RMA cells in vitro and in vivo. Methods： The recombination vector pcDNA3.1-E7 carrying wild type HPV 16 E7 was identified by sequencing. The recombination vector pcDNA3.1-E7 was transfected into mouse lymphadenoma cell line RMA by liposome, and the monoclonal cells transfected stably were obtained by antibiotics G418 sieving and limiting dilution assay. RT-PCR method was used to detect the expression of HPV 16 E7 mRNA in RMA-E7 cells. The growth of RMA cells and RMA-E7 cells cultured in vitro was tested by Cell Count Kit-8. RMA-E7 cells and RMA cells were subcutaneously inoculated in syngeneic mice respectively, the tumor size was measured by sliding caliper twice a week, and the E7 protein expression in tumor tissue of mice was detected by Western blot after tumor formation. The kinetics of cytolytic activity of E7 specific T cells in tumor-bearing mice was measured by LDH kit. Results： Sequencing of recombination vector showed the target gene which was inserted into the recombinant was correct, and RMA-E7 cells expressing E7 protein stably were obtained by limited dilution assay. There were no obvious differences in morphous and growth velocity between RMA cells and RMA-E7 cells in vitro. RMA-E7 cells grew in syngeneic mice were significantly slower than RMA cells. The E7 protein was ex- pressed stronger in RMA-E7 cells in vivo than in vitro. The cytolytic ability of ET-specific CTL was activated at the early stage, reached the maximum at the middle stage, and lost at the end stage. RMA-E7 cells isolated from the tumor-bearing mice were more resistant to E7-specific CTL killing than RMA-E7 cells cultured in vitro. Conclusion： The E7 protein expression has no obvious influence on growth of RMA-E7 cells in vitro, and can suppress growth of RMA-E7 cells in vivo. The activity curve of E7 specific CTL approximately presents ＂bell＂ shape.

关键词： cytotoxic T lymphocyte human papilloma virus (HPV) type 16 E7 immune tolerance