Inactivation of the Wnt/β-catenin signaling contributes to the epithelial barrier dysfunction induced by sodium oxalate in canine renal epithelial cells

High oxalate consumption has been recognized as a risk factor for renal calcium oxalate stones in companion animals (dogs and cats). However, the cellular signaling involved in oxalate-induced dysfunction in renal tubular epithelial cells remains not fully elucidated. In this study, Mardin–Darby canine kidney (MDCK) cells, an epithelial cell line derived from canine kidney tubule, were tested for cell proliferation activity and barrier function after being exposed to sodium oxalate (NaOx). Further, the involvement of Wnt/β-catenin in NaOx-induced renal epithelial barrier dysfunction was evaluated. MDCK cells treated with NaOx exhibited reduction in cell proliferation and migration. Besides, NaOx exposure led to a decrease in transepithelial electrical resistance and an increase in paracellular permeability. The deleterious effects of NaOx on epithelial barrier function were related to the suppressed abundance of tight junction proteins including zonula occludens, occludin, and claudin-1. Of note, protein levels of β-catenin and phosphorylated (p)-β-catenin (Ser552) in MDCK cells were repressed by NaOx, indicating inhibitory effects on Wnt/β-catenin signaling. An inhibition of glycogen synthase kinase-3β (GSK-3β) by SB216763 enhanced the abundance of β-catenin and p-β-catenin (Ser552), and protected against epithelial barrier dysfunction in NaOx-treated MDCK cells. The results revealed a critical role of Wnt/β-catenin signaling in the epithelial barrier function of MDCK cells. Activation of Wnt/β-catenin signaling might be a potential therapeutic target for the treatment of oxalate-linked renal stones.     

ADF-IL-2基因重组卡介苗的构建及免疫保护效果

构建柔嫩艾美尔球虫ADF基因和鸡IL-2基因的重组卡介苗,并研究其免疫保护性.将柔嫩艾美尔球虫的ADF基因和鸡的IL-2基因串联后连接到至穿梭表达载体pMV261和整合表达载体pMV361中电转化卡介苗,制备重组卡介苗pMV261-ADF-IL-2和pMV361-ADF-IL-2,将构建的基因重组卡介苗分别以不同免疫途径接种雏鸡,研究其对柔嫩艾美耳球虫的攻毒的免疫保护效果.结果显示,成功构建重组rBCG pMV261-ADF-IL-2和pMV361-ADF-IL-2,且rBCG滴鼻免疫组的免疫效果较好,其抗球虫指数分别为176.08和172.89.结果表明,构建的基因重组卡介苗pMV261-ADF-IL-2和pMV3 61-ADF-IL-2对柔嫩艾美尔球虫卵囊的攻击具有一定的保护作用.     

2011年中国北方H9N2亚型禽流感病毒分离株HA基因进化分析及受体结合性检测

为了解近期中国北方禽源H9N2亚型禽流感病毒(AIV)流行规律及分子遗传进化特征,本实验对2011年在中国北方家禽中分离到的11株H9N2亚型AIV通过RT-PCR扩增病毒的HA基因片段,进行测序及遗传进化分析,并对这些病毒的受体结合性进行了检测.结果表明:11株H9N2亚型AIV分离株的HA基因在HA1和HA2的氨基酸裂解位点均为PSRSSR/GLF基序,符合低致病性病毒株氨基酸序列特征.多数病毒HA潜在糖基化位点为8个,所有分离株病毒的受体结合位点226位均为L,经红细胞受体结合性试验验证表明这些病毒均同时具有α-2,3和α-2,6受体结合特性,表明目前北方地区流行的H9N2亚型AIV具有感染哺乳动物的潜在威胁.本研究结果对加强H9N2病毒的分子流行病学监测具有重要意义.     

猪β2m四聚体前体链重组表达载体的构建和表达

为构建猪β2m轻链四聚体前体链,通过PCR定点突变技术,将猪主要组织相容性复合体Ⅰ类分子轻链β2m羧基端69位苯丙氨酸(Phe)突变为半胱氨酸(Cys),然后将突变体基因插入到pET-28a载体,转化BL21细胞,经诱导后SDS-PAGE检测蛋白质表达情况。结果显示,β2m突变体基因全长297 bp,编码98个氨基酸,其中羧基端69位Phe突变为Cys。突变体基因经诱导表达后进行SDS-PAGE检测,结果显示蛋白质表达大小为10.6 ku。本研究构建了猪β2m轻链突变体基因重组表达载体,并成功表达了目的蛋白,为今后构建四聚体链奠定基础。     

H9N2亚型禽流感流行分析及疫苗毒株的筛选

本研究旨在筛选新型免疫原性好、广谱的优秀 H9亚型禽流感病毒疫苗株。通过 H9N2分子流行病学调查对分离得到的37株禽流感 H9N2亚型病毒进行 HA 基因分子遗传进化及其关键位点分析,利用交叉 HI 效价及鸡胚中和试验分析毒株间毒力及其相关性,制备疫苗用毒株进行免疫攻毒保护试验。结果显示,从37株病毒中选出12株病毒作为代表,分为3个分支,同分支之间交叉 HI 效价及中和效价最高。HI 试验及鸡胚中和试验均显示同分支毒株间的相关系数高。免疫攻毒保护试验显示同分支疫苗株安全性高。表明我国禽流感 H9N2亚型情况复杂,且疫苗保护效果与疫苗株的抗原匹配性密切相关,生产多种分支 H9疫苗株联合更有利于控制 H9型禽流感,     

单端孢霉烯族毒素生物脱毒技术研究进展

单端孢霉烯族毒素在粮食和饲料中污染严重,主要是T-2毒素和脱氧雪腐镰刀烯醇(DON)等,其毒性作用强,对人和动物产生严重危害.因此,单端孢霉烯族毒素的脱毒方式逐渐成为研究热点,尤其是生物脱毒.很多微生物被证实能够降解霉菌毒素,包括细菌、真菌和酵母菌,可将毒素降解成低毒或无毒的产物.论文以生物脱毒为出发点,从细菌、真菌和...     

Effects of the sodium‐glucose cotransporter 2 (SGLT2) inhibitor velagliflozin,a new drug with therapeutic potential to treat diabetes in cats

Sodium‐glucose cotransporter 2 (SGLT2) inhibitors are used in the treatment of human diabetics. They increase glucose excretion and correct hyperglycemia. We examined the investigational SGLT2 inhibitor velagliflozin in two groups of six neutered adult obese cats (equal gender distribution). Placebo (Pl) or drug (D; 1 mg/kg) was administered for 35 days. Routine blood examinations, fructosamine, beta‐hydroxybutyrate (BHB), nonesterified fatty acids (NEFA), glucagon, adiponectin, and leptin were measured before and after treatment, also water intake, and urinary electrolytes, glucose, and volume. Indirect calorimetry, an intravenous glucose tolerance test (IVGTT; 0.8 g/kg) and insulin tolerance test (IVITT) were conducted. All cats tolerated treatment well. Significant changes with D included a decrease in the respiratory exchange ratio, an increase in cholesterol, a small increase in albumin, and a rise in BHB and NEFA. Glucose clearance was unaltered, although less insulin was secreted during the IVGTT (p = .056) suggesting improved insulin sensitivity. IVITT was unchanged. Treatment did not affect glucagon, leptin, or adiponectin. Water intake, urine output, urinary glucose excretion, and the glucose/creatinine ratio but not urinary electrolytes were significantly higher post‐D. We conclude that velagliflozin is a promising drug, which increases urinary glucose excretion in cats and could thereby be beneficial for the treatment of hyperglycemia.     

一株鸭源禽流感病毒（H9N2）全基因序列分析及其排毒规律的研究

为研究一株鸭源H9N2亚型禽流感病毒(JN1株)全基因组的分子特征和遗传进化情况以及感染雏鸭后的排毒规律,对其生物学特性、全基因组序列以及遗传进化进行了分析,通过点眼滴鼻方式感染3周龄健康雏鸭,利用建立的实时荧光定量PCR(RT-PCR)检测鸭的排毒规律,并测定血清抗体效价。JNI株的鸡胚半数感染量为10~(-7.54)/0.2mL,鸡胚最小致死量的平均死亡时间为72h,静脉致病指数为0.266,抗原相关系数为0.47。系统发育分析表明,JN1株的HA与CK/HK/G9/97和DK/HK/Y280/97在同一分支上,NA、M、NP、PA、PB1、PB2基因与SH/F/98同源性较高,NS基因则与H5N1亚型禽流感病毒进化关系较近。HA的裂解位点为PSRSSR↓GL,存在6个潜在糖基化位点,在234位的氨基酸残基为L,NA基因颈部存在缺失。雏鸭在感染该病毒后,饮食正常,精神良好,3~17d均有排毒,咽拭子和泄殖腔棉拭子排毒规律基本一致,分别在第3天和第13天出现排毒高峰。该H9N2亚型禽流感病毒属欧亚大陆分支,为低致病性禽流感,病毒可通过呼吸道和泄殖腔向外界排毒,排毒时间较长。     

盐胁迫对碱茅幼苗叶片内源激素、NAD激酶及Ca2+-ATPase的效应

为了研究盐渍生境对盐生植物碱茅Puccinellia chinampoensis体内部分信号分子的影响,采用不同浓度NaCl溶液处理碱茅植株,对幼苗叶片中部分信号分子及相关蛋白测定和分析.结果发现,低盐下碱茅无胁迫反应,高盐胁迫下,碱茅脱落酸(ABA)含量升高,生长素(IAA)、赤霉素(GA)含量均下降,细胞分裂素(ZR)含量趋于稳定;碱茅NAD激酶(NADKase)活性升高,提高代谢调节能力;同时Ca2 -ATPase活性增加,是对盐胁迫引起Ca2 浓度升高而做出的胁迫反应.表明了碱茅通过胞间信号相对含量的变化,调节NADKase活性、Ca2 -ATPase活性,提高耐盐能力.     

家蚕滞育卵浸酸后易溶性蛋白的表达差异分析

探明浸酸解除家蚕卵滞育的关联蛋白,可为改进和提高蚕种的人工孵化技术提供理论依据。以经2.5℃冷藏45 d的家蚕品种"7.湘×9.芙"滞育卵为材料,采用双向凝胶电泳和图像分析技术,比较浸酸与未浸酸蚕卵易溶性蛋白的差异变化。在未浸酸蚕卵的双向电泳图谱中共检测到523个蛋白质斑点,其中特异性蛋白质斑点41个;浸酸卵共检测到526个蛋白质斑点,其中特异性蛋白质斑点44个。浸酸与未浸酸蚕卵能相互匹配的蛋白质斑点有482对,匹配率91.897%。经对比分析,浸酸活化后的蚕卵蛋白出现了一些特异性的蛋白质斑点,或某些蛋白质斑点在含量上出现了差异变化。