iNOS-GC-cGMP signaling activation might be involved in vascular hyporeactivity in endotoxemic rats

AIM: To evaluate the activation of inducible nitric oxide synthase (iNOS)-guanylate cyclase(GC)-cyclic guanosine monophosphate(cGMP) signaling on vascular hyporeactivity in endotoxemic rats. METHODS: Twenty-four SD rats were randomly divided into 4 groups as follows: sham operation group (sham group), lipopolysaccharide group(LPS group), LPS+polymyxin B group (LPS+PMX-B group) and polymyxin B group (PMX-B group). Cannulation of the carotid artery was performed to record mean arterial blood pressure (MABP). The levels of plasma NO, iNOS and TNF-α were detected. The tension of the thoracic aortic rings was measured by a biological analytical system. RESULTS: Compared with sham group, MABP in LPS group was significantly lower (P<0.01), whereas MABP in LPS+PMX-B group was significantly higher than that in LPS group (P<0.05), and no statistical difference of MABP between PMX-B group and sham group was observed (P>0.05). The plasma levels of NO and iNOS in LPS group were significantly higher than those in sham group and LPS+PMX-B group (P<0.01). The contraction of isolated thoracic aortic rings stimulated by phenylephrine and the relaxation response by acetylcholine in LPS group were significantly lower than those in sham group (P<0.01), whereas those in LPS+PMX-B group were significantly improved (P<0.01). The vascular hyporeactivity to vasoconstrictors was completely reversed by pretreatment either with aminoguanidine, a selective iNOS inhibitor, or with methylene blue, an inhibitor of NO-sensitive GC. CONCLUSION: The iNOS-GC-cGMP signaling activation might be involved in vascular hyporeactivity in LPS-induced endotoxemic rats. Polymyxin B partly reverses the vascular hyporeactivity to vasoconstrictors by reducing the level of serum TNF-α, which may be mediated by the iNOS-GC-cGMP signal pathways to attenuate the overexpression of iNOS and NO production.     

Protective effect of folate and vitamin B12 on Alzheimer-like pathological changes in brain of aged rats with hyperhomocysteinemia

AIM:To explore the effect of hyperhomocysteinemia on the Alzheimer-like pathological changes in the brain of aged rat. METHODS:The rats were treated with homocysteine(Hcy) by intravenous injection through vena caudalis with or without a simultaneous supplementation of folate and Vitamin B₁₂(FB) in the drinking water for 28 weeks. The distribution and aggregation of tau protein were detected by Bielschowsky silver staining. Western blotting was used to determine the protein levels of tau phosphorylation, glycogen synthase kinase 3β(GSK-3β) and protein phosphatase 2A(PP2A). RESULTS:Treatment with Hcy induced hyperhomocysteinemia in aged rats and resulted in the formation of neurofibrillary tangles in the brain. Supplementation of FB effectively reduced the tangles. Hyperhomocysteinemia also induced Alzheimer-like tau hyperphosphorylation at multiple sites(Ser^199/202and Ser³⁹⁶). Hyperhomocysteinemia activated GSK-3β and inhibited the activity of PP2A. Supplementation of FB alleviated the changes of tau and the activities of GSK-3β and PP2A. CONCLUSION:Supplementation of FB ameliorates the hyperhomocysteinemia-induced Alzheimer-like pathological changes of tau protein possibly through regulating the activity of GSK-3β and PP2A in aged rats.     

奶牛性控冻精和常规冻精活力对比试验分析

试验观测对比奶牛性控冻精和常规冻精解冻后精子存活时间,为进一步把握输精时间,提高受胎率提供依据。试验结果表明:性控冻精刚解冻时活力较高,为0.5级以上,但活力下降快、存活时间较短,解冻后6h活力降低为零;常规冻精刚解冻时活力在0.4～0.5级之间,但存活时间较长,活力下降速度慢,解冻后8h活力降低为零。所以性控冻精解冻后,要求输精技术员尽快输精。     

山羊子宫内膜基质细胞对上皮细胞IL-18分泌活性的调节作用

为了探究性腺激素作用下山羊子宫内膜上皮细胞（EEC）中白细胞介素-18（IL-18）的分泌活性,以及子宫内膜基质细胞（ESC）对其分泌活性的调节作用。基于套皿共培养永生化山羊EEC和ESC为体外研究模型,通过ELISA和Western blot方法检测E2和/或P4对EEC中IL-18分泌水平的作用以及ESC对该作用的影响。ELISA结果显示,E2较无激素对照组可显著增强单独培养EEC中IL-18的分泌水平（P〈0.05）,P4单独或与E2联合作用则进一步增强其分泌活性（P〈0.01）;EEC与ESC共培养模式下,ESC细胞对性激素作用下EEC中IL-18分泌活性具有显著的抑制作用。Western blot结果显示,在单独培养或与ESC共培养模式下,EEC培养上清液中IL-18均以相对分子质量大小18 300的具有生物活性的蛋白形式存在。本研究结果表明,山羊ESC对性腺激素作用下EEC中IL-18的分泌活性具有重要调节作用。     

绵羊自然感染贝氏莫尼茨绦虫对小肠黏膜免疫相关细胞的数量变化

运用组织学和组织化学染色法,分别对自然感染贝氏莫尼茨绦虫的成年绵羊（感染组）与正常成年绵羊（对照组）的小肠各肠段的上皮内淋巴细胞、浆细胞、杯状细胞和嗜酸粒细胞的分布和数量变化进行了比较研究。结果显示：感染组小肠各段上皮内淋巴细胞、浆细胞、杯状细胞和嗜酸粒细胞数量均显著高于对照组,其中感染组十二指肠、空肠和回肠的上皮内淋巴细胞数量分别比对照组增加了169.11%、230.38%和233.42%（P〈0.01）;嗜酸粒细胞数量分别比对照组增加了116.78%、123.87%和164.51%（P〈0.01）;浆细胞数量分别比对照组增加了127.34%、72.97%和328.26%（P〈0.01）;杯状细胞数量分别比对照组增加了33.40%、41.42%和133.17%。对照组和感染组上皮内淋巴细胞和杯状细胞从十二指肠到回肠均逐渐减少,相反,对照组和感染组嗜酸粒细胞数量从十二指肠到回肠均逐渐增多,对照组小肠固有层浆细胞数量从十二指肠到空肠增加,空肠到回肠减少,感染组小肠固有层浆细胞数量从十二指肠到回肠均逐渐增多;对照组十二指肠、空肠、回肠的上皮内淋巴细胞、上皮内杯状细胞、浆细胞和嗜酸粒细胞均差异极显著（P〈0.01）;感染组十二指肠、空肠、回肠的上皮内杯状细胞、浆细胞和嗜酸粒细胞均差异极显著（P〈0.01）,感染组上皮内淋巴细胞空肠与回肠差异极显著（P〈0.01）,但十二指肠与空肠差异不显著（P〉0.05）。研究结果表明,贝氏莫尼茨绦虫感染成年绵羊后,成年绵羊通过特异性黏膜免疫细胞上皮内淋巴细胞增生加强细胞免疫水平,浆细胞增生加强体液免疫水平,同时还通过非特异性黏膜免疫细胞,嗜酸粒细胞和杯状细胞的增生进一步加强黏膜免疫水平以抵抗贝氏莫尼茨绦虫对绵羊的感染。可见绵羊可以通过黏膜免疫相关细胞增生加强局部免疫力以监视虫体免疫逃逸来抵抗寄生虫的感染。     

30～38周龄产蛋鸡理想氨基酸模式的研究

本试验旨在通过4个剂量-反应试验,研究30～38周龄产蛋鸡对赖氨酸(Lys)、蛋氨酸(Met)、色氨酸(Trp)和苏氨酸(Thr)4种必需氨基酸(AA)的需要量及其比例.每个试验均采用360只海兰灰产蛋鸡,设5个处理,分别采食5个AA梯度水平的试验饲粮,每个处理6个重复,每个重复12只鸡.采用蛋公鸡回肠食糜法测定饲料原料的标准回肠可消化(SID)值.试验预试期14 d,正试期56 d.结果表明:海兰灰产蛋鸡的产蛋率、蛋重、日产蛋量和料蛋比与SIDLys、SID Met、SID Trp和SID Thr的摄入水平均有较好的折线回归关系;以日产蛋量为自变量(x),AA摄入水平为依变量(Y)分别建立的回归方程为:YSID Lys=51.60-0.041 6(682.5-x),R2=0.96;YsID Met=51.43-0.085(314.1-x),R2 =0.95;YSID Trp =51.17-0.155 4(163.0-x),R2=0.97;YsID Thr=52.85-0.057 6(499.5-x),R2=0.93.在饲粮粗蛋白质12.8％的条件下,以日产蛋量为参照指标确定的30 ～ 38周龄产蛋鸡的SID Lys、SID Met、SID Trp和SID Thr需要量分别为684、313、171和506 mg/(只·d).以日产蛋量为指标建立的30～38周龄产蛋鸡理想SID AA模式为:Lys:Met:Trp:Thr=100∶46∶24∶73.     

蛋鸡J亚群禽白血病病毒3′非编码区序列特征分析

蛋用型鸡虽在试验条件下可以感染ALV-J,但自然感染时很少引起发病.然而,近年来在中国蛋鸡群中ALV-J发病严重,本研究对2008年以来ALV-J蛋鸡分离株的3′非编码区(3′UTR)进行了系统的分析.结果表明,89.5％(17/19)的ALV-J蛋鸡分离株的3′UTR出现了205 bp的缺失,94.7％(16/17)ALV-J蛋鸡分离株保留了完整的E元件.84.2％(16/19)的ALV-J蛋鸡分离株的U3区序列与肉鸡分离株的相似性低于92.5％,所有转录调控元件高度保守.ALV-J蛋鸡分离株的E元件和U3区均出现了多个碱基的突变.这些结果表明,ALV-J蛋鸡分离株的E元件和U3区正在迅速的演化,明显不同于肉鸡分离株的序列特征.这些发现有助于更好地了解ALV-J引起蛋鸡群发病的致病机理.     

三种猪繁殖与呼吸综合征疫苗免疫效果评价

为研究不同猪繁殖与呼吸综合征病毒(PRRSV)疫苗的的免疫特性,分别采用PRRSV变异株(JXA1-R)弱毒疫苗、经典PRRSV(VR 2332)弱毒疫苗、变异株(JXA1)灭活疫苗,免疫接种PRRSV抗原和抗体阴性的健康断奶仔猪,免疫接种后70d用PRRSV变异株强毒攻毒,ELISA检测血清中PRRSV特异的抗体水平及IFN-γ、IL 2、IL 4、IL 8、IL 10的水平,并进行临床症状和肺部病理组织学观察和评分。结果表明,VR 2332、JXA1-R弱毒疫苗对免疫猪攻毒保护效果差异不显著,均为63.6%(7/11),JXA1灭活疫苗免疫攻毒保护效果较差,为36.4%(4/11)。试验还发现病毒感染猪血清中细胞因子IFN-γ和IL-10的比值可以作为评价疫苗免疫效果的一个指标。     

狐狸源致病性维氏气单胞菌的分离鉴定及耐药性分析

本研究通过生物学特性、16S rRNA基因和4种等看家基因的序列测定与分析,对分离自北京动物园死亡狐狸病变组织的1株细菌进行鉴定,并对其进行小鼠致病力试验及药物敏感性试验。结果显示:该细菌为革兰氏阴性杆菌,培养特性、理化反应与维氏气单胞菌温和生物型基本相同;根据16S rRNA基因和dnaJ、rpoD、gyrB、cpn60等看家基因的系统进化及其同源性分析,确定所分离的细菌与维氏气单胞菌属于同一系统发育分支,各基因同源性最高分别为97.6%～99.9%。分离菌株对CD-1小鼠有较强的致病作用,其半数致死量为2.8×105cfu/只;对氧氟沙星、头孢他啶等14种抗菌药物敏感,对青霉素G、克林霉素等9种抗菌药物耐药。     

马流感病毒血凝素基因甲病毒复制子重组表达质粒的构建及其免疫效力评价

为评价马流感病毒(EIV)HA基因核酸免疫效果,本研究以甲病毒复制子载体pSFV1CS分别构建了表达EIV H3N8亚型的美洲型和欧洲型HA基因的重组真核表达质粒。并将其转染293T细胞,经间接免疫荧光鉴定表明HA基因获得表达;以重组质粒免疫的BALB/c鼠能够检测到特异性抗体产生,而且HI抗体水平持续升高,同时小鼠体内IFN-γ、IL-4分泌水平也有所升高。攻毒后小鼠表现轻度临床症状,但病毒分离和RT-PCR均未检测到病毒。上述结果表明,该重组质粒pSFV1CS-EIV-HA具有良好的免疫原性并且可以诱导免疫动物产生较高免疫应答的能力。