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Oral immunization of mice with a probiotic Lactobacillus casei constitutively expressing the α-toxoid induces protective immunity against Clostridium perfringens α-toxin

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VIRULENCE 2019年第1期10卷 166-179页

作者： Gao, xuwen Ma, Yingying Wang, Zhuo Bai, Jing Jia, Shuo Feng, Baohua Jiang, Yanping Cui, Wen tang, Lijie Li, Yijing Wang, Li Xu, Yigang Northeast Agr Univ Coll Vet Med Harbin Heilongjiang Peoples R China China Minist Agr Key Lab Anim Pathogen Biol Northeastern Sci Inspect Stn Harbin Heilongjiang Peoples R China Northeast Agr Univ Heilongjiang Key Lab Anim Dis Control & Pharmaceu Harbin Heilongjiang Peoples R China

Clostridium perfringens alpha-toxin is one of the major virulence factors during C. perfringens infection, causing hemolysis of erythrocytes in various species. Here, genetically engineered Lactobacillus casei (pPG-alpha/L. casei 393) constitutively expressing the toxoid of C. perfringens alpha-toxin was generated and its immunogenicity in mice for induction of protective immunity against the alpha-toxin was evaluated via oral immunization. The alpha-toxoid was constitutively expressed by pPG-alpha/L. casei 393 without a specific inducer, as confirmed by western blotting, laser confocal microscopy, and flow cytometry. In an experiment on BALB/c mice to evaluate the oral immunogenicity of pPG-alpha/L. casei 393, significant levels of a specific secretory IgA (sIgA) antibody in the intestinal mucus and feces and an IgG antibody in the serum of the probiotic vaccine group were detected after booster immunization (p < 0.05) as compared with the pPG/L. casei 393 and PBS control groups. These antibodies effectively neutralized C. perfringens natural alpha-toxin. Moreover, significantly higher levels of cytokines IL-2, IL-4, IL-10, IL-12, IL-17, and interferon (IFN) gamma in the serum and increased proliferation of spleen lymphocytes obtained from mice orally immunized with pPG-alpha/L. casei 393 were detected. With a commercial C. perfringens type A inactivated vaccine as a control, immune protection provided by the probiotic vaccine against C. perfringens alpha-toxin was evaluated, and 90% and 80% protection rates were observed, respectively. Therefore, strain pPG-alpha/L. casei 393 effectively elicited mucosal, humoral, and cellular immunity, suggesting that pPG-alpha/L. casei 393 is a promising candidate for development of a vaccine against C. perfringens alpha-toxin.

关键词： Clostridium perfringens alpha-toxoid genetically engineered Lactobacillus casei oral immunization

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基于模糊PID控制的船用胎架高度联控方法研究

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江苏科技大学学报（自然科学版） 2021年第1期35卷 58-65页

作者：刘金锋王镇景旭文唐明明曹旭武董建伟赵晨华江苏科技大学机械工程学院镇江212100 陕西柴油机重工有限公司兴平713100

胎架作为船舶分段建造变形控制的重要工装,但不具有联合调控与自适应控制能力,无法快速响应分段建造质量问题,为此本文提出基于模糊PID控制的船用胎架高度联控方法.首先,基于船舶分段拟合曲面建立胎架高度补偿模型,为胎架的自适应调控... 详细信息

胎架作为船舶分段建造变形控制的重要工装,但不具有联合调控与自适应控制能力,无法快速响应分段建造质量问题,为此本文提出基于模糊PID控制的船用胎架高度联控方法.首先,基于船舶分段拟合曲面建立胎架高度补偿模型,为胎架的自适应调控提供依据;其次,基于分段变形范围和受压载荷构建胎架间关联机制,达到胎架系统的动态联调;然后,利用模糊PID控制算法实现胎架高度的自适应调控,达到抑制分段外板变形的目的;最后,通过智能胎架系统对本方法进行验证分析,并与常规PID对比分析,表明本文方法具有较好的稳定性.

关键词：船舶分段建造智能胎架高度调控模糊PID

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Metabolomic study of natrin-induced apoptosis in SMMC-7721 hepatocellular carcinoma cells by ultra-performance liquid chromatography-quadrupole/time-of-flight mass spectrometry

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INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES 2019年第0期124卷 1264-1273页

作者： Lu, Shiyin Lu, Rigang Song, Hui Wu, Jinxia Liu, xuwen Zhou, Xiaoling Yang, Jianqing Zhang, Hongye tang, Chaoling Guo, Hongwei Hu, Jian Mao, Guifu Lin, Hanmei Su, Zhiheng Zheng, Hua Guangxi Med Univ Pharmaceut Coll Nanning Peoples R China Liuzhou Tradit Chinese Med Hosp Dept Pharm Liuzhou Peoples R China Guangxi Inst Food & Drug Control Nanning Peoples R China Liuzhou Tradit Chinese Med Hosp Dept Gastroenterol Liuzhou Peoples R China Guangxi Tradit Chinese Med Univ Affiliated Hosp 1 Gynaecol Nanning Peoples R China Guangxi Med Univ Med Sci Res Ctr Nanning Peoples R China

Natrin, a new member of the cysteine-rich secretory protein (CRISP) family purified from the snake venom of Naja naja atra, has been demonstrated to have anticancer activity. However, the underlying molecular mechanisms need further elucidation. In this study, MU was used to evaluate cell viability. Apoptotic cells were analyzed by employing a transmission electron microscope (TEM). Metabolomic study of the metabolic perturbations caused by natrin-induced apoptosis in differentiated SMMC-7721 cells was performed for the first time by using integrative ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q/IDF MS). To investigate the possible mechanism in the mitochondria] pathway of natrin-induced apoptosis, we measured apoptosis-related mRNA changes using real-time fluorescent quantitative PCR (FQ-PCR). Cell proliferation was significantly inhibited after treatment with natrin in a dose dependent manner. Principal component analysis (PCA) and partial least squares-discriminate analysis (PLS-DA) clearly demonstrated that metabolic profiles were affected by natrin. The results of multivariate statistical analysis showed that a total of 13 metabolites were characterized as potential biomarkers highly implicated in natrin-induced apoptosis, which corresponded to fluctuations of five pathways, including sphingolipid metabolism, fatty acid biosynthesis, fatty acid metabolism, glycerophospholipid metabolism and glycosphingolipid biosynthesis. Furthermore, natrin-induced apoptosis showed an increase in the Bax/Bcl-2 ratio in the mitochondrial pathway compared with controls. This study illustrated that rapid and holistic cell metabolomics combining molecular biological approaches might be a powerful tool for evaluating the underlying mechanisms of natrin-induced apoptosis, which would help to deepen specific insights into the anti-hepatoma mechanisms of natrin and facilitate the clinical application of natrin in the future.

关键词： Metabolomics Natrin Hepatocellular carcinoma (HCC) Apoptosis UPLC-QtrOF MS

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Protective Effect of sRAGE on Fetal Development in Pregnant Rats with Gestational Diabetes Mellitus

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CELL BIOCHEMISTRY AND BIOPHYSICS 2015年第2期71卷 549-556页

作者： tang, xuwen Qin, Qingxin Xie, Xiaobin He, Ping Guangzhou Women & Childrens Med Ctr Dept Obstet Guangzhou 510623 Guangdong Peoples R China First Peoples Municipal Hosp Guangzhou Dept Endocrinol Guangzhou 510180 Guangdong Peoples R China Guangzhou Med Univ Dept Pathol Guangzhou 510180 Guangdong Peoples R China

To investigate the protective effect of secretory receptor for advanced glycation endproducts (sRAGE) on the fetal development using rat model of gestational diabetes mellitus (GDM). The model of pregnant rats with intrauterine hyperglycemia was established by intraperitoneal injection of 25 mg/kg streptozotocin (STZ). Rats with established GDM were randomly grouped, and the pregnant rats in the experimental group were subsequently injected with recombinant sRAGE protein (5 mg/kg, in 0.2 mL PBS) at tail vein every 24 h, while the rats in control group were injected with the same dosage of albumin solution. Blood glucose, serum levels of advanced glycation endproducts (AGEs), and levels of RAGE protein in brain and heart tissues of pregnant rats were measured at 3, 13, and 19 days postconception. At 19 days fetuses were delivered by cesarean section, number of fetuses, their weight and placental weights were recorded, and fetal malformations and defects were analyzed visually and pathologically. The expression level of RAGE, NOX2, MCP-1, p65, VCAM-1, and VEGF mRNA in placenta was evaluated by real-time PCR. p65 protein localization was detected by immunohistochemistry in fetal brain and heart tissue sections. We analyzed the correlation between AGEs and RAGE level and the development of fetal rats, and the protective effect of blocking AGEs-RAGE pathway on the fetal development in the rat model of GDM was investigated. (1) The concentration of blood glucose and AGEs in serum of pregnant rats with GDM was significantly higher than in control group (p < 0.05), with strong correlation between blood glucose and levels of AGEs (r = 0.693, p < 0.05). (2) While both the number of fetuses and placental wet weight in pregnant rat model of GDM were similar to control group, pups from GDM group exhibited higher incidence of developmental abnormalities and higher average weight (p < 0.05). sRAGE treatment slightly but not significantly reduced the probability of the fetal develo

关键词： Gestational diabetes mellitus Secretory receptor for advanced glycation endproducts Protection Fetal development

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分泌型晚期糖基化终产物受体对妊娠糖尿病胚胎保护的实验研究

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西部医学 2017年第10期29卷 1356-1360页

作者：秦庆新汤栩文谢晓斌卢建文广州市第一人民医院内分泌科广东广州510180 广州市妇女儿童医疗中心广东广州510120 广州医科大学广东广州510180

目的研究分泌型晚期糖基化终产物受体(sRAGE)在实验动物罹患妊娠糖尿病(GDM)时的表达差异及对子鼠发育的影响。方法采用给SD孕鼠空腹一次性腹腔注射链脲佐菌素(STZ)25mg/kg的方法建立宫内高血糖孕鼠模型。STZ注射后对小鼠随机分为实验... 详细信息

目的研究分泌型晚期糖基化终产物受体(sRAGE)在实验动物罹患妊娠糖尿病(GDM)时的表达差异及对子鼠发育的影响。方法采用给SD孕鼠空腹一次性腹腔注射链脲佐菌素(STZ)25mg/kg的方法建立宫内高血糖孕鼠模型。STZ注射后对小鼠随机分为实验组和对照组,每间隔24小时,实验组孕鼠行尾静脉注射重组sRAGE蛋白(5mg/kg,0.2ml PBS)即为sRAGE组,对照组注射相同剂量的白蛋白溶液。分别于第3、13、19天检测孕鼠血糖、血清AGEs及脑、心脏组织中的RAGE蛋白水平。最后一日剖宫取胎,剥离胎盘,检测胎盘RAGE、NOX2、MCP-1、VCAM-1的mRNA表达水平,探讨血清中AGEs、RAGE表达水平与胚胎发育的关系。结果实验组孕鼠血清中血糖和AGEs的浓度显著高于对照组(P<0.05),相关性分析显示血糖与血清中AGEs水平呈显著正相关(r=0.693,P<0.05);与正常对照组相比,实验组孕鼠的胎盘、脑及心脏组织中活性氧产生,炎症相关基因的mRNA和RAGE蛋白水平显著升高,而sRAGE处理可以部分降低这些活性氧产生、炎症相关基因的表达和RAGE的表达。对照组孕鼠胎盘、脑以及心脏组织中的RAGE蛋白表达极弱,实验组在3种组织中的RAGE表达则明显增强。在sRAGE注射干预后,RAGE蛋白表达与实验组相比显著降低,仍高于对照组。结论 sRAGE静脉注射可显著降低孕鼠子代组织中NF-κB的活性,通过调控相关细胞因子发挥对胚胎的保护作用。

关键词：妊娠糖尿病分泌型型晚期糖基化终产物受体胚胎保护

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CNTs/CNFs/GCE电化学传感器的制备及其对木犀草素的检测

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广西科技大学学报 2025年第2期36卷 118-127页

作者：朱旭文郭双李春婷张丽文程昊唐婷范广西糖资源绿色加工重点实验室(广西科技大学) 广西柳州545006 广西柳州螺蛳粉工程技术研究中心(广西科技大学) 广西柳州545006 广西科技大学生物与化学工程学院广西柳州545006 杭州微念品牌管理有限公司浙江杭州310000

以碳纳米管(carbon nanotubes,CNTs)和聚丙烯腈(polyacrylonitrile,PAN)为前驱体,通过静电纺丝得到纳米纤维,后经碳化得到CNTs/CNFs复合材料。使用扫描电子显微镜(scanning electron microscope,SEM)、X射线衍射(X-ray diffraction,XRD)、能量色谱仪(energy dispersive spectroscopy,EDS)和电化学阻抗谱(electrochemical impedance spectroscopy,EIS)对其形貌和组成进行了表征;以滴涂法将此材料修饰到玻碳电极(glassy carbon electrode,GCE)表面,考察木犀草素在该复合电极上的电化学行为;最后将此修饰电极应用到金银花露中木犀草素的测定。结果表明:此修饰电极在检测木犀草素的试验中展现出良好的重复性、重现性和抗干扰能力;当木犀草素浓度在3~10μmol/L时,CNTs/CNFs/GCE修饰电极响应电流值与木犀草素浓度呈良好的线性关系,检测限为0.18μmol/L;用此复合电极检测金银花露中的木犀草素,加标回收率在96.069%~97.623%之间,相对标准偏差小于5.0%。

关键词：木犀草素碳纳米管静电纺丝法电化学加标回收率

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金属有机多面体基包覆型微球制备及CO2环加成反应

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南京工业大学学报（自然科学版） 2020年第4期42卷 536-542页

作者：吕保帅周洁蒙旭雯张竹修汤吉海崔咪芬乔旭南京工业大学化工学院材料化学工程国家重点实验室江苏南京211800

采用原位合成法在铜基海藻酸-壳聚糖-SiO2(Cu-ALG-CS-SiO2)内部生长金属有机多面体(MOP),制备MOP基包覆型微球MOP-ALG-CS-SiO2。通过X线衍射(XRD)、N2吸附-脱附、透射电子显微镜(TEM)等方法对MOP-ALG-CS-SiO2的晶形、孔径和形貌进行分... 详细信息

采用原位合成法在铜基海藻酸-壳聚糖-SiO2(Cu-ALG-CS-SiO2)内部生长金属有机多面体(MOP),制备MOP基包覆型微球MOP-ALG-CS-SiO2。通过X线衍射(XRD)、N2吸附-脱附、透射电子显微镜(TEM)等方法对MOP-ALG-CS-SiO2的晶形、孔径和形貌进行分析并考察MOP-ALG-CS-SiO2催化环氧化合物与CO2环加成制备环状碳酸酯的反应性能。结果表明:MOP-ALG-CS-SiO2具有无定形的介孔结构,MOP在MOP-ALG-CS-SiO2中高度分散,解决了MOP基复合材料中普遍存在的稳定性和分散性不好的问题。MOP-ALG-CS-SiO2在催化环氧丙烷与CO2环加成反应中,环氧丙烷转化率为84.1%、碳酸丙烯酯选择性为99.7%,催化性能优于MOP、MOP-ALG-CS和HKUST-1。此外,MOP-ALG-CS-SiO2在催化其他类型环氧化物与CO2环加成反应中均表现出较高的催化活性和产物选择性。

关键词：金属有机多面体(MOP) 原位合成介孔微球环加成反应

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Factors associated with blooms of cyanobacteria in a large shallow lake, China

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ENVIRONMENTAL SCIENCES EUROPE 2018年第1期30卷 1-15页

作者： Li, Di Wu, Naicheng tang, Song Su, Guanyong Li, xuwen Zhang, Yong Wang, Guoxiang Zhang, Junyi Liu, Hongling Hecker, Markus Giesy, John P. Yu, Hongxia Nanjing Univ Sch Environm State Key Lab Pollut Control & Resource Reuse Nanjing 210046 Jiangsu Peoples R China Jiangsu Environm Monitoring Ctr Nanjing 210036 Jiangsu Peoples R China Univ Kiel Dept Hydrol & Water Resources Management D-24118 Kiel Germany Chinese Ctr Dis Control & Prevent Natl Inst Environm Hlth Beijing 100021 Peoples R China Nanjing Univ Sci & Technol Sch Environm & Biol Engn Jiangsu Key Lab Chem Pollut Control & Resources R Nanjing 210094 Jiangsu Peoples R China Nanjing Normal Univ Sch Environm Nanjing 210023 Jiangsu Peoples R China Wuxi Environm Monitoring Ctr Wuxi 214000 Jiangsu Peoples R China Univ Saskatchewan Toxicol Ctr Saskatoon SK S7N 5B3 Canada Univ Saskatchewan Sch Environm & Sustainabil Saskatoon SK S7N 5C3 Canada Univ Saskatchewan Dept Vet Biomed Sci Saskatoon SK S7N 5B3 Canada Univ Saskatchewan Toxicol Ctr Saskatoon SK S7N 5B3 Canada Michigan State Univ Dept Zool E Lansing MI 48824 USA Michigan State Univ Ctr Integrat Toxicol E Lansing MI 48824 USA Univ Hong Kong Sch Biol Sci Hong Kong Hong Kong Peoples R China

Background: Eutrophication of freshwater systems can result in blooms of phytoplankton, in many cases cyanobacteria. This can lead to shifts in structure and functions of phytoplankton communities adversely affecting the quality of drinking water sources, which in turn impairs public health. Relationships between structures of phytoplankton communities and concentrations of the toxicant, microcystin-leucine-arginine (MC-LR), have not been well examined in large shallow lakes. The present study investigated phytoplankton communities at seven locations from January to December of 2015 in Tai Lake, and relationships between structures and diversities of phytoplankton communities and water quality parameters, including concentrations of MC-LR and metals, were analyzed. Results: A total of 124 taxa of phytoplankton were observed, and the predominant taxa were Microcystis sp. and Dolichospermum flos-aquae of Cyanophyta and Planctonema sp. of Chlorophyta. The greatest diversities of phytoplankton communities, as indicated by species richness, Simpson, Shannon-Wiener, the Berger and Parker, and the Pielou evenness indices, were observed in spring. Furthermore, productivity of phytoplankton was significantly and negatively correlated with diversities. These results demonstrated that Simpson, Shannon-Wiener, the Berger and Parker, and the Pielou evenness indices of phytoplankton communities were significantly related to trophic status and overall primary productivity in Tai Lake. In addition, temperature of surface water, pH, permanganate index, biochemical oxygen demand, total phosphorus, arsenic, total nitrogen/total phosphorous ratio, and MC-LR were the main factors associated with structures of phytoplankton communities in Tai Lake. Conclusion: The present study provided helpful information on phytoplankton community structure and diversity in Tai Lake from January to December of 2015. Our findings demonstrated that Simpson, Shannon-Wiener, the Berger and Parker, and the

关键词： Eutrophication Nutrients Phosphorus Nitrogen Phytoplankton productivity Diversity Microcystin-LR

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Dual priming oligonucleotide (DPO)-based real-time RT-PCR assay for accurate differentiation of four major viruses causing porcine viral diarrhea

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MOLECULAR AND CELLULAR PROBES 2019年 47卷 101435-000页

作者： Jia, Shuo Feng, Baohua Wang, Zhuo Ma, Yingying Gao, xuwen Jiang, Yanping Cui, Wen Qiao, Xinyuan tang, Lijie Li, Yijing Wang, Li Xu, Yigang Northeast Agr Univ Heilongjiang Key Lab Anim Dis Control & Pharmaceu Coll Vet Med Harbin Heilongjiang Peoples R China China Minist Agr Key Lab Anim Pathogen Biol Northeastern Sci Inspect Stn Harbin Heilongjiang Peoples R China

Currently in China, porcine epidemic diarrhea virus (PEDV), transmissible gastroenteritis virus (TGEV), porcine rotavirus (PoRV), and porcine deltacoronavirus (PDCoV) are the major causes of porcine viral diarrhea, and mixed infections in clinics are common, resulting in significant economic losses in pig industry. Here, a dual priming oligonucleotide (DPO)-based multiplex real-time SYBR Green RT-PCR assay were developed for accurately differentiating PEDV, TGEV, PoRV, and PDCoV in clinical specimens targeting the N gene of TGEV, PEDV, and PDCoV, and the VP7 gene of PoRV. Results showed that the DPO primer allowed a wider annealing temperature range (40-65 degrees C) and had a higher priming specificity compared to conventional primer, in which more than 3 nucleotides in the 3'- or 5'-segment of DPO primer mismatched with DNA template, PCR amplification efficiency would decrease substantially or extension would not proceed. DPO-based multiplex real-time RT-PCR method had analytical detection limit of 8.63 x 10(2) copies/mu L, 1.92 x 10(2) copies/mu L, 1.74 x 10(2) copies/mu L, and 1.76 x 10(2) copies/mu L for PEDV, TGEV, PoRV, and PDCoV in clinical specimens, respectively. A total of 672 clinical specimens of piglets with diarrhea) symptoms were collected in Northeastern China from 2017 to 2018 followed by analysis using the assay, and epidemiological investigation results showed that PEDV, TGEV, PoRV, and PDCoV prevalence was 19.05%, 5.21%, 4.32%, and 3.87%, respectively. The assay developed in this study showed higher detection accuracy than conventional RT-PCR method, suggesting a useful tool for the accurate differentiation of the four major viruses causing porcine viral diarrhea in practice.

关键词： Porcine viral diarrhea Dual priming oligonucleotide (DPO) Real-time RT-PCR assay Accurate differentiation

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Paecilomyces variotii extracts（ZNC） enhance plant immunity and promote plant growth

Paecilomyces variotii extracts（ZNC） enhance plant immunity...

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中国植物病理学会2019年学术年会

作者： Lu Chongchong Liu Haifeng Jiang Depeng Wang Lulu Jiang Yanke tang Shuya Hou xuwen Han Xinyi Liu Zhiguang Zhang Min Chu Zhaohui Ding Xinhua State Key Laboratory of Crop Biology Shandong Provincial Key Laboratory for Biology of Vegetable Diseases and Insect PestsShandong Agricultural University Shandong Pengbo Biotechnology Co. LTD National Engineering Laboratory for Efficient Utilization of Soil and Fertilizer Resources College of Recourses and EnvironmentShandong Agricultural University

The crude extract of the endophytePaecilomyces variotii known as ZhiNengCong（ZNC）has function of promoting plant growth and enhancing disease resistance and is widely used in *** study aims to evaluate the molecular mechanisms of plant growth promotion and disease *** generated transcriptome profiles from ZNC-treated seedlings using RNA *** function of salicylic acid（SA） in ZNC-mediated immunity was examined using SA biosynthesis and signaling pathway *** concentrations of nitrogen（N） and phosphorus（P） in seedlings under ZNC treatment were *** effect of ZNC on the level of the hormone auxin in roots was tested using transgenic plants containing DR5:*** exhibited ultrahigh activity in promoting plant growth and enhancing disease resistance,even at concentrations as low as 1-10 ng/*** induced ROS accumulation,callose deposition,and expression of PR *** biosynthesis and signaling pathways were required for the ZNC-mediated defense ***,in improving plant growth,ZNC increased the level of auxin in root tips and regulated the absorption of N and *** to these results,ZNC is a highly effective plant elicitor that promotes plant growth by inducing auxin accumulation at the root tip at low concentrations and enhances plant disease resistance by activating the SA signaling pathway at high concentrations.

关键词： Plant elicitors Plant growth promotion Plant endophyte Disease resistance Auxin Salicylic acid

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