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猪胞内劳森菌抗体间接ELISA检测方法的建立及应用

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中国预防兽医学报 2024年第1期46卷 55-60,69页

作者：裴艳艳张梦琳许浒相丽润张洪亮彭金美王倩田志军周国辉中国农业科学院哈尔滨兽医研究所动物疫病防控全国重点实验室黑龙江哈尔滨150069

猪增生性肠病(PPE),通常被称为猪回肠炎(PI),由胞内劳森氏菌(LI)感染引起。为建立检测LI抗体的间接ELISA方法,本研究构建LI外膜蛋白基因(LI0841)的重组表达质粒p ET28a-LI0841,经测序无误后转化BL21(DE3)宿主菌,经IPTG诱导表达,采用SDS-... 详细信息

猪增生性肠病(PPE),通常被称为猪回肠炎(PI),由胞内劳森氏菌(LI)感染引起。为建立检测LI抗体的间接ELISA方法,本研究构建LI外膜蛋白基因(LI0841)的重组表达质粒p ET28a-LI0841,经测序无误后转化BL21(DE3)宿主菌,经IPTG诱导表达,采用SDS-PAGE检测重组蛋白LI0841蛋白(rLI0841)的表达形式,经Ni柱纯化后采用western blot鉴定其反应原性。SDS-PAGE结果显示,在32 ku处出现目的条带,且其主要以可溶性形式表达;western blot结果显示,r LI0841能够与兔LI多克隆抗体特异性反应,表明纯化的r LI0841具有较强的反应原性,可作为包被抗原用于建立间接ELISA检测方法,经各反应条件优化初步建立LI抗体的间接ELISA检测方法。各反应条件的优化结果显示,4.38 ng/孔的r LI0841以4℃过夜包被最佳;血清最佳稀释度为1∶100,37℃反应0.5 h;羊抗猪Ig G-HRP最佳稀释度为1∶10000,37℃作用0.5 h;TMB底物37℃显色15 min。利用建立的间接ELISA方法检测猪繁殖与呼吸障碍综合征病毒、猪伪狂犬病病毒、副猪嗜血杆菌、猪链球菌、传染性胸膜肺炎放线杆菌及经美国Biostone PPE抗体检测试剂盒检测为阳性的猪血清,评估该方法的特异性;将LI阳性血清2倍倍比稀释(1∶100~1∶51200)后,采用本研究建立的间接ELISA方法检测,评估该方法的敏感性;以同一批次和不同批次包被的酶标板分别检测5份不同LI抗体水平的猪血清,评估该方法的重复性。结果显示,该方法除能检测到LI阳性血清外,其余相关病原的阳性血清均为阴性,特异性较强;LI阳性血清1∶800稀释时检测结果仍为阳性,敏感性较高;对5份不同抗体水平的LI阳性血清的批内、批间重复性试验的变异系数均小于10%,重复性较好。利用该ELISA方法与美国Biostone公司PPE抗体检测试剂盒同时检测104份临床猪血清样品,比较二者的检测结果,并计算二者的符合率;采用建立的间接ELISA方法检测黑龙江、吉林等地区413份临床猪血清样品,分析LI在上述地区的流行状况。结果显示,两种方法的阳性符合率为90.91%,阴性符合率为91.84%,总符合率为91.35%;413份临床猪血清样品中LI的阳性检出率为59.81%(247/413),表明LI在黑龙江、吉林等地区的猪群中普遍存在。本研究建立了检测LI抗体的间接ELISA方法,该方法特异性强、敏感性高、重复性与准确性均较好,为临床PI血清流行病学调查提供技术支持。

关键词：胞内劳森氏菌间接ELISA 抗体检测 LI0841重组蛋白

Effect of Sheng Xue Ning Tablets on Renal Anemia in Patients Subject to Maintenance Hemodialysis and Safety Evaluation: A Multi-setting Prospective Randomized Study

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Current Medical Science 2020年第2期40卷 327-331页

作者： Xiao-jing TANG Shu RONG Chang-lin MEI zhao-hui NI Geng-ru JIANG Wei-jie YUAN Nian-song wang Zhi-yong guo Jun MA Hai-dong yan Li-ming ZHANG Department of Nephrology Changzheng Hospitalthe Second Military Medical UniversityShanghai 20003China Department of Nephrology Shanghai General HospitalShanghai Jiao Tong Universiry School of MedicineShanghai 200080China Department of Nephrology Renji HospitalJiaotong UnivrsiryShanghai 200003China Department of Nephrology Xin/ua HospitalJiaotong UniversitShanghai 200092China Deparment of Nephrology.the Sixth People's Hospital Jiaotong UniversityShanghai 200233China Department of Nephrology Changhai Hospitalthe Second Military Medical UniversityShanghai 200433China Deparment of Nephrology Tongji HospitalTongji UniversityShanghai 200065China Deparment of Nephrology East HospialTongjt UniversiryShanghai 200120China Department of Nephrology Zhabei District Cenral HospitalChangzhen HospitalShanghai 200070China

This study compared Sheng Xue Ning(SXN)tablets with ferrous succinate(FS)tablets in terms of their efficacy for the treatment of iron-deficient renal anemia and safety in patients subject to maintenance hemodialysis(MHD).A total of 94 patients undergoing MHD were randomly assigned to an experiment group(receiving oral SXN tablcts,SXN group)and a control group(orally given FS tablets,FS group)and followed up for 12 ***(EPO)was used in both *** eficacy was assessed by detecting the subsequent changes in hemoglobin(Hb),serum iron(SI),SF and transferrin saturation(TSAT).At the 12th week,Hb and TSAT levels in both groups were significantly increased compared to those in the screening period(P<0.05).However,no significant difference in Hb and TSAT was found between the two *** average weekly EPO dosage used was lower in SXN group than in FS group(P<0.05)at the 10th week and the 12th *** study showed that SXN tablets can effectively ameliorate renal anemia and keep iron metabolism stable in MHD patients,and its efficacy is virtually close to that of FS ***,SXN tablets can reduce the dosage of EPO and have a good safety profile.

关键词： hemodialysis renal anemia oral iron supplements

Influencing Factors for Erectile Dysfunction of Young Adults with No Response to PDE5i

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Current Medical Science 2021年第3期41卷 529-534页

作者： Jin wang yan-li wang Yao HE Sen LI You-peng ZHANG yong-biao CHENG Jia-lun guo Lin SUN Yu XIN yong TANG zhao-hui ZHU Han-qing ZENG Department of Urology Union HospitalTongji Medical CollegeHuazhong University of Science and TechnologyWuhan 430022China

Erectile dysfunction(ED)is a common male *** orally-administered phosphodiesterase type 5 inhibitors(PDE5 inhibitors)are now recognized as the primary pharmacological treatment method for ED,20%-30%of the patients treated with PDE5 inhibitors exhibit no significant *** study aims to investigate the influencing factors of ED in young adults with no response to PDE5 *** patients who would take PDE5 inhibitors were included and investigated with a *** with no response to PDE5 inhibitors(tadalafil and sildenafil)served as study group,and those with response to PDE5 inhibitors as control *** Chi square test and logistic regression analysis were applied to find the potential influencing *** total,378 ED patients were ***-three(24.6%)cases were non-responsive to PDE5 inhibitors,and the remaining 285(75.4%)responded to PDE5 *** multiple logistic regression analysis,we found that history of drinking(OR=3.152;95%CI 1.672-6.975),spousal noncooperation(OR=2.994;95%CI 1.589-5.638),number of fixed sex partners(OR=0.358;95%CI 0.132-0.651),duration of ED(OR=3.356;95%CI 1.352-8.333),and depression(OR=3.689;95%CI 1.579-8.979)could be the influencing factors for ED patients’non-response to PDE5 *** conclusion,history of drinking,spousal noncooperation,number of fixed sex partner,long duration of ED,and depression could be the influencing factors for ED patients'non-response to PDE5 *** and doctors should pay attention to these factors.

关键词： erectile dysfunction PDE5 inhibitors influencing factor

丹皮酚对大鼠卵母细胞体外成熟体系氧化应激的调控

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菏泽医学专科学校学报 2024年第1期36卷 11-16页

作者：张春强田荆华王守福赵辉李岩郭新庆韦岩王海蓉刘胜薛兰菏泽医学专科学校山东菏泽274000

目的研究中药提取物丹皮酚对大鼠卵母细胞体外培养体系氧化应激的改善作用,旨在确定丹皮酚改善氧化应激作用的最佳浓度,以期优化卵母细胞的体外培养体系,提高生产效率。方法在卵母细胞的体外培养体系中分别添加2 mmol/L、4 mmol/L、6 ... 详细信息

目的研究中药提取物丹皮酚对大鼠卵母细胞体外培养体系氧化应激的改善作用,旨在确定丹皮酚改善氧化应激作用的最佳浓度,以期优化卵母细胞的体外培养体系,提高生产效率。方法在卵母细胞的体外培养体系中分别添加2 mmol/L、4 mmol/L、6 mmol/L、8 mmol/L、10 mmol/L的丹皮酚,研究体外成熟后卵母细胞的成熟质量和氧化应激水平。结果添加丹皮酚均能提高卵母细胞的成熟率(P<0.05)。添加8 mmol/L的丹皮酚获得卵母细胞的GVBD率、受精率和囊胚率高于其他组(P<0.05);添加6 mmol/L、8 mmol/L和10 mmol/L组的活性氧水平较低,与2 mmol/L、4 mmol/L组具有极显著差异(P <0.01)。添加丹皮酚能够显著提高成熟卵母细胞BCL-2基因和蛋白的表达水平,降低BAX基因和Caspase-3基因和蛋白的表达水平(P<0.01)。添加8 mmol/L、10 mmol/L组较其它组差异极显著(P<0.01)。结论在大鼠卵母细胞体外成熟液中添加8 mmol/L的丹皮酚可以提高卵母细胞的体外成熟的生产效率和质量,且能够有效改善氧化应激水平、降低卵母细胞的凋亡。

关键词：丹皮酚大鼠卵母细胞体外成熟体系氧化应激

miR-103-3p targets Ndel1 to regulate neural stem cell proliferation and differentiation

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Neural Regeneration Research 2022年第2期17卷 401-408页

作者： Wen Li Shan-Shan wang Bo-Quan Shan Jian-Bing Qin He-yan zhao Mei-Ling tian hui He Xiang Cheng Xin-Hua Zhang guo-Hua Jin Department of Human Anatomy Institute of NeurobiologyNantong UniversityNantongJiangsu ProvinceChina Key Laboratory of Neuroregeneration of Jiangsu and Ministry of Education NantongJiangsu ProvinceChina Co-Innovation Center of Neuroregeneration Nantong UniversityNantongJiangsu ProvinceChina

The regulation of adult neural stem cells(NSCs) is critical for lifelong neurogenesis. MicroRNAs(miRNAs) are a type of small, endogenous RNAs that regulate gene expression post-transcriptionally and influence signaling networks responsible for several cellular processes. In this study, mi R-103-3 p was transfected into neural stem cells derived from embryonic hippocampal neural stem cells. The results showed that mi R-103-3 p suppressed neural stem cell proliferation and differentiation, and promoted apoptosis. In addition, mi R-103-3 p negatively regulated Nud E neurodevelopment protein 1-like 1(Ndel1) expression by binding to the 3′ untranslated region of Ndel1. Transduction of neural stem cells with a lentiviral vector overexpressing Ndel1 significantly increased cell proliferation and differentiation, decreased neural stem cell apoptosis, and decreased protein expression levels of Wnt3 a, β-catenin, phosphor-GSK-3β, LEF1, c-myc, c-Jun, and cyclin D1, all members of the Wnt/β-catenin signaling pathway. These findings suggest that Ndel1 is a novel mi R-103-3 p target and that mi R-103-3 p acts by suppressing neural stem cell proliferation and promoting apoptosis and differentiation. This study was approved by the Animal Ethics Committee of Nantong University, China(approval No. 20200826-003) on August 26, 2020.

关键词： apoptosis canonical Wnt pathway differentiation MiR-103-3p Ndel1 neural stem cells neurogenesis proliferation

Enhancing storage performance of P2-type Na_(2/3)Fe_(1/2)Mn_(1/2)O_(2)cathode materials by Al_(2)O_(3)coating

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Transactions of Nonferrous Metals Society of China 2022年第1期32卷 262-272页

作者： Yi-jiao CHANG Guang-hui XIE yong-mao ZHOU Jie-xi wang Zhi-xing wang Hua-jun guo Bian-zheng YOU guo-chun yan School of Metallurgy and Environment Central South UniversityChangsha 410083China

The P2-type Na_(2/3)Fe_(1/2)Mn_(1/2)O_(2)materials were synthesized by an ultrasonic spray pyrolysis followed by solid-state sintering *** structures,morphologies and electrochemical performances of Na_(2/3)Fe_(1/2)Mn_(1/2)O_(2)materials were characterized thoroughly by means of X-ray diffractometer,scanning electron microscope and electrochemical charge/discharge ***,a thin layer of Al_(2)O_(3),which was formed on the surface of Na_(2/3)Fe_(1/2)Mn_(1/2)O_(2),can enhance the storage performance by preventing the formation of Na_(2)CO_(3)·H_(2)O,which is believed to enhance the electrochemical performances of Na_(2/3)Fe_(1/2)Mn_(1/2)O_(2)*** facile surface modification method may pave a way to synthesize advanced cathode materials for sodium-ion batteries.

关键词： Al_(2)O_(3)coating air-stability spray pyrolysis sodium-ion batteries iron-manganese oxide

Growth of Zinc Blende GaAs/AlGaAs Radial Heterostructure Nanowires by a Two-Temperature Process

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Chinese Physics Letters 2011年第3期28卷 140-143页

作者： guo Jing-Wei HUANG hui REN Xiao-Min yan Xin CAI Shi-Wei guo Xin HUANG yong-Qing wang Qi ZHANG Xia wang Wei Key Laboratory of Information Photonics and Optical Communications(Ministry of Education) Beijing University of Posts and TelecommunicationsBeijing 100876

Zinc blende structure GaAs/AlGaAs core-multishell nanowires(NWs)are grown on a GaAs(111)B substrate by a two-temperature process using an Au-catalyzed vapor-liquid-solid mechanism and metal organic chemical vapor deposition,***-free radial heterostructure NWs are *** can be concluded that the NWs are grown with the main contributions from the direct impingement of the precursors onto the alloy droplets and little from adatom *** results indicate that the droplet acts as a catalyst rather than an adatom *** photoluminescence spectra reveal that the grown NWs have much higher optical efficiency than bare GaAs NWs.

关键词： GaAs/AlGaAs nanowires Zinc

Identification and comparative analysis of the major chemical constituents in the extracts of single Fuzi herb and FuziGancao herb-pair by UFLC-IT-TOF/MS

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Chinese Journal of Natural Medicines 2014年第7期12卷 542-553页

作者： yanG yan YIN Xin-Juan guo hui-Min wang Ru-Lin SONG Rui tian Yuan ZHANG Zun-Jian Key Laboratory of Drug Quality Control and Pharmacovigilance China Pharmaceutical UniversityMinistry of Education State Key Laboratory of Natural Medicines China Pharmaceutical University

AIM: The aim of this work was to establish a specific and sensitive method to comprehensively investigate and compare chemical constituents of Fuzi-Gancao herb pair(FG), consisting of Aconitum carmichaelii Debeaux(Fuzi, Chinese) and Roast Radix Glycyrrhizae(Glycyrrhiza glabra L., Gancao, in Chinese) and Fuzi alone to explore the underlying interaction mechanism of FG. METHOD: An ultra-fast liquid chromatography-ion trap/time-of-flight mass spectrometry(UFLC/MS-IT-TOF) method using diazepam as internal standard was developed for the identification and semi-quantitative analysis of the phytochemical constituents of Fuzi and FG. Chromatographic separation was achieved on a UFLC column using a gradient program with 40 mmol?L-1 ammonium acetate and acetonitrile as the mobile phase. RESULTS: Fifty-one of the sixty compounds, including forty-five C19-diterpenoid alkaloids and six C20-diterpenoid alkaloids were tentatively identified in the extracts of Fuzi and FG through accurate mass measurements and fragmentation patterns. Comparing the contents of these alkaloids in these two extracts, it was found that the diester-diterpenoid alkaloids(DDAs) and the alkylolamine-diterpenoid alkaloids(ADAs) were increased, while the monoester-diterpenoid alkaloids(MDAs) were decreased in the extracts of FG. CONCLUSION: This work provided comprehensive information for the quality control of Fuzi preparations, and the further investigation on the compatibility mechanisms of FG.

关键词： Aconitum carmichaelii Radix Rhizoma Glycyrrhizae Diterpenoid alkaloids UFLC-IT-TOF/MS

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焦棓酸对猪链球菌体外抑菌效果试验

动物医学进展 2024年第7期45卷 76-81页

作者：高婷王艳君王雅雯袁芳艳刘泽文刘威杨克礼郭锐李畅黎璐田永祥周丹娜湖北省农业科学院畜牧兽医研究所湖北武汉430064 华中农业大学动物医学院湖北武汉430070 老河口牧原农牧有限公司湖北老河口441000

研究焦棓酸对猪链球菌的体外抗菌活性及对生物被膜的抑制作用,并初步探讨其抑菌机制。采用96孔微量稀释法测定常用抗菌药物及焦棓酸对猪链球菌的最小抑菌浓度和最小杀菌浓度;平板活菌计数法和分光光度计测定焦棓酸对猪链球菌作用的时间... 详细信息

研究焦棓酸对猪链球菌的体外抗菌活性及对生物被膜的抑制作用,并初步探讨其抑菌机制。采用96孔微量稀释法测定常用抗菌药物及焦棓酸对猪链球菌的最小抑菌浓度和最小杀菌浓度;平板活菌计数法和分光光度计测定焦棓酸对猪链球菌作用的时间-杀菌曲线;采用结晶紫染色法检测焦棓酸对猪链球菌生物被膜形成的影响;通过荧光定量PCR从转录水平分析焦棓酸对猪链球菌生长与生物被膜形成相关基因表达的情况。结果显示,猪链球菌2型HB菌株对磺胺异恶唑、磺胺甲恶唑耐药严重,焦棓酸对猪链球菌2型HB菌株的最小抑菌浓度为32μg/mL,最小杀菌浓度为64μg/mL,并且焦棓酸对猪链球菌的抑制作用呈剂量依赖性;焦棓酸在16μg/mL能显著抑制猪链球菌2型HB菌株生物被膜形成的能力;另外,经焦棓酸处理后,发现猪链球菌的生长、生物被膜形成、毒力相关基因ftsZ、stK、mrp、fbps、gapdh、ccpA、luxS下调表达。综上所述,焦棓酸可为耐药猪链球菌在临床上感染的治疗提供新思路。

关键词：焦棓酸猪链球菌抗菌活性生物被膜