猪戊型肝炎病毒ORF2片段克隆及原核表达

设计套式引物，内套引物含有BamHⅠ／XhoⅡ酶切位点，采用RT-PCR技术扩增猪戊型肝炎病毒（UEV）结构蛋白基因ORF2部分片段，长度为634bp，将其克隆于PGEX-T载体，测序结果表明插入的片段属于戊型肝炎病毒ORF2部分，将该片段插入PproEXHTb表达载体，经酶切鉴定证实获得了重组表达质粒。将重组质粒转化大肠杆菌BL21，经1mmol／L IPTG诱导，得到表达，表达蛋白分子量为27Ku，以包涵体形式存在。Western blot分析表明，该蛋白可以与猪戊型肝炎阳性血清反应，表明该蛋白具有良好的反应原性。     

猪肉风味的影响因素

风味是评价猪肉品质的重要指标之一,它受很多因素影响。本文论述了饲养管理方式、日粮来源与营养素、贮藏条件、季节等因素对猪肉风味的影响,讨论了建立客观的风味评定体系的迫切性。     

Effect of diet with high fats and cholesterol on PPARα gene expression and body cholesterol level in mice

AIM:To investigate the effects of dietary fats and cholesterol on liver PPARα gene expression and body cholesterol (Chol) level in C57BL/6J mice. METHODS:The animals (n=75) were randomly divided into five groups and respectively received formula mash for 6 weeks. RESULTS:As compared to Chol diet, Chol+PUFA diet produced significantly higher liver cholesterol (P<0.01), serum total cholesterol (TC), focusing on HDL-C. While Chol+MUFA diet resulted in unchanged serum TC and lower liver cholesterol (P<0.01). Chol+SFA diet rsulted in higher liver cholesterol (P<0.01) and serum TC, focusing on LDL-C. Furthermore, Chol+PUFA diet increased the mRNA and protein content of PPARα (P<0.01) in liver, while Chol+MUFA and Chol+SFA diets decreased the mRNA content of PPARα significantly (P<0.01). CONCLUSIONS:These results indicated that addition of fats containing PUFA to diet high in cholesterol increased PPARα mRNA and protein expression, addition of fats containing MUFA or SFA to diet high in cholesterol decreased PPARα mRNA expression. The change of PPARα gene expression may further affect body cholesterol level.     

Effect of endogenous H2S on pulmonary hypertension during acute lung injury induced by LPS and its interaction with NO

AIM: To investigate the effect of H2S on pulmonary artery hypertension during acute lung injury induced by LPS and the interaction between the systems of hydrogen sulfide (H2S)/cystathionine-β-lyase (CSE) and nitric oxide (NO)/nitric oxide synthase (NOS) in this process. METHODS: Seventy-two adult male rats were randomly divided into four groups: control group, LPS group, LPS+L-NAME group and LPS+propargylglycine (PPG) group. Mean pulmonary artery pressure (mPAP) of each rat was examined at 2 h, 4 h, 6 h and 8 h after treatment. H2S and NO contents in plasma, NO content, iNOS, cNOS and CSE activity in lung were measured at 4 h or 8 h after treatment, respectively. Expression of iNOS in lung tissue was also detected by immunohistochemistry technique, and the injury of lung was evaluated with morphological changes under microscope. RESULTS: LPS could induce severe lung injury, and mPAP, NO content, iNOS activity and its protein expression in LPS group significantly increased, but cNOS activity, H2S content and CSE activity decreased compared with those of control group. Administration of L-NAME before LPS could attenuate the changes induced by LPS. Pre-administration of PPG, a CSE inhibitor, exacerbated the injury by LPS, but there was no prominent variation in cNOS activity. CONCLUSION: Reduced endogenous H2S could increase pulmonary artery hypertension during acute lung injury induced by LPS. There is a negative effect between H2S/CSE system and NO/NOS system in this process.     

Exogenous carbon monoxide protects against the lung injury induced by ischemia-reperfusion of hind limbs in rats

AIM: To study the mechanism of protective effect of exogenous carbon monoxide (CO) in the lung injury induced by ischemia-reperfusion (IR) of hind limbs in rats. METHODS: Thirty-two SD rats were randomly divided into 4 groups: control, control+CO, IR and IR+CO. A rat model of ischemia in hind limbs and the reperfusion lung injury was made. The rats in IR+CO and control+CO groups were exposed to air containing 2.5×10-8 CO for 1 h before reperfusion or the corresponding control time point, while the other two groups were exposed to the routine air. The lung tissue structure, polymorphonuclear leukocyte (PMN) count, wet-to-dry weight ratio (W/D), malondialdehyde (MDA) content and the animal survival rate were observed. The carboxyhemoglobin (COHb) levels in artery blood were detected with CO-oximeter and the expression of intercellular adhesion molecule-1 (ICAM-1) in the lung was detected by Western blotting. RESULTS: Compared to control, the animal mortality, lung PMNs number, W/D, MDA content and ICAM-1 expression were all significantly increased in IR group. Compared with the IR group, the blood COHb level was significantly increased and the animal mortality, lung PMNs number, W/D, MDA content and ICAM-1 expression were all significantly decreased in IR+CO group. CONCLUSION: These data suggest that exogenous CO attenuate limb IR-induced lung injury by down-regulatiny ICAM-1 expression and suppressing PMN sequestration in the lung following limb IR in rats.     

茚酮肟醚衍生物的合成与生物活性

通过傅－克反应及环化、肟化、成醚等反应,合成了10个未见文献报道的茚酮肟醚类化合物,其结构均经1H NMR、IR、LC/MS及元素分析确证。初步生物活性测试结果表明,所合成的化合物具有明显的杀菌活性或杀虫活性,如在100 mg/L时,化合物 V1 对小麦白粉病菌Erysiphe graminis 的抑制率为 95.6%, V5 对粘虫Mythimna separata的致死率为100%。     

人为干扰对典型草原土壤侵蚀影响的价值评价

基于不同干扰条件下典型草原风洞试验的研究结果,计算了风蚀所造成的各种损失(养分、有机碳的损失及废弃地的形成等等),在此基础上,定量评价了人为干扰对土壤侵蚀过程的影响。研究结果表明,禁牧可以明显减小由风蚀所导致的草原生态系统的价值损失,而开垦和放牧则会增加其价值损失(包括速效养分、有机碳和废弃地形成)。总的来看,不同风速下不同生态系统的价值损失速率由大到小的排列顺序为:新开垦农田>农田>自由放牧草原>2003年禁牧草原>1987年禁牧草原;同时,风速越大不同干扰类型之间的价值损失差值越大。     

尼卡巴嗪和氨丙啉单独及配合应用对鸡球虫病的疗效

目的研究尼卡巴嗪和氨丙啉单独及配合应用对鸡球虫病的疗效。方法将15日龄鸡人工感染柔嫩艾美耳球虫,以鸡平均增重率、盲肠病变记分、粪便记分和抗球虫指数等指标为参数,综合评价尼卡巴嗪和氨丙啉单独及配合应用的抗球虫效果。结果尼卡巴嗪左旋咪唑 大蒜素 维生素K组抗球虫指数为189。氨丙啉 乙氧酰胺甲酯 乙胺嘧啶 磺胺喹啉组抗球虫指数为184.5,这两组药均具有良好的抗球虫效果。氨丙啉 乙氧酰胺苯甲酯组,氨丙啉 乙胺嘧啶组及氨丙啉药物对照组抗球虫指数分别为162.8,163.2,和160.5;尼卡巴嗪 乙氧酰胺苯甲酯组和单用尼卡巴嗪组抗球虫疗效较差,AC I分别为142.7和146.4。结论尼卡巴嗪配合左旋咪唑、大蒜和维生素K产生了增效作用;用氨丙啉配合乙氧酰胺苯甲酯、乙胺嘧啶和磺胺喹啉也可达到增效目的,经两药单用提高了疗效。     

猪CTSD全长cDNA的克隆和表达分析

以人CTSD mRNA全长序列为基序,在db EST库中搜索同源性大于80%、重叠大于40个碱基的猪EST下载经Seqman软件装配后,利用RT-PCR扩增到猪CTSD基因部分cDNA序列,进一步通过RACE技术获得cDNA全长（GenBank登录号为DQ018727）,猪CTSD基因cDNA全长2032 bp包括93 bp 5′UTR区、706 bp 3′UTR区和1 233bp ORF,编码410个氨基酸残基。其编码区与人、鼠、牛、羊CTSD编码区同源性分别为87.9%、78.2%、86.6%和85.0%,其氨基酸序列与人、鼠、牛、羊氨基酸序列同源性分别为86.6%、80.1%、86.0%和84.7%。疏水性分析和信号肽预测发现猪CTSD氨基酸序列存在至少7个疏水性区域,信号肽位点为第1-20个氨基酸残基。在NCBI进行Blast P搜索结果显示猪CTSD氨基酸结构中含有Asn（Asparagine,天门冬酰胺）结构域,与天冬氨酸蛋白酶3D结构同源性高达99.7%,具有真核生物天冬氨酸蛋白酶的典型结构。以猪多组织RNA池为模板,以1026 bp部分cDNA序列作为杂交探针进行Northern杂交,得到一条2000 bp左右特异条带,从而验证了RACE产物为全长cDNA并揭示该基因只有一个转录本。为了研究猪CTSD基因在体内不同组织内表达的特点,采用半定量RT-PCR对CTSD基因在猪10个组织中的表达进行研究,结果表明在10个组织中均有表达,且表达量与-βactin内参接近。     

Effects of immune-enhancing parenteral nutrition on nutritional status,immunity and inflammatory responses in postoperative patients with portal hypertension surgery

AIM:This study was conducted to evaluate the effects of postoperative immune-enhancing parenteral nutrition on the nutritional statue,immune function,and inflammatory responses in patients undergoing portal hypertension surgery. METHODS: This study was designed as a prospective,randomized and controlled clinical trial. Forty-two patients undergoing portal hypertension surgery were randomly assigned to receive either an immune-enhancing parenteral nutrition(adding glutamine and recombinant human growth hormone,n=22) or an isocaloric and isonitrogenous control standard parenteral nutrition(n=20) for seven days. Parenteral nutrition was initiated 3 days after surgery. Blood samples were obtained on day 0,3,and 10. Host nutritional statue was evaluated by measuring levels of prealbumin and transferrin,immunity was observed by measuring levels of CD4+ cells,CD8+ cells,CD4+/CD8+,IgG,IgM and IgA,and the inflammatory responses was determined by assessing IL-2,TNF-α and C-reactive protein(CRP) levels. RESULTS:On postoperative day 10,among patients receiving an immune-enhancing parenteral nutrition,prealbumin,transferrin,CD4+ cells,CD4+/CD8+,IgG and IL-2 levels were significantly higher than those in control group,and TNF-α and CRP concentrations were significantly decreased（P<0.05）. CONCLUSION:Postoperative administration of immune-enhancing parenteral nutrition in patients undergoing portal hypertension surgery can improve postoperative nutritional statue and immune function,and relieve inflammatory response.