白三叶和黑麦草品种在成都地区的引种试验

对引自百绿集团的白三叶4个品种、多花黑麦草2个品种和多年生黑麦草2个品种在成都地区进行引种试验。结果表明：各品种均能适应当地气候条件,其中白三叶Sustain和C6151再生好,产量高,分别为88350kg／hm2和84450kg／hm2;多花黑麦草再生速度快,产量高,青绿期长,营养价值高;均可在四川盆地大面积推广。多年生黑麦草再生慢,产量低,且越夏性能差,可在高原牧区推广。     

家蚕肾形卵的荧光差异显示分析

Liang等建立的mRNA的荧光差异显示法 (flourescentdifferentialdisplay,FDD)具有高效、安全、有用信息多等特点。利用这一方法在mRNA水平上探讨了肾形卵 (ki)与正常卵在初期胚胎阶段的基因表达 ,回收了差异片段 10条 ,并对其差异进行了验证。其中之一的片段序列分析表明为低分子量热激蛋白基因 (smallheatshockgene)。     

猪细胞因子cDNA表达文库的构建及其基因的克隆鉴定

为克隆和研究猪细胞因子及相关基因,应用建库试剂盒成功构建了猪细胞因子cDNA表达文库。采取健康猪外周血及淋巴结,分离单个核细胞,经LPS PHA联合刺激不同时间后,提取总RNA。将各组样品混合,分离纯化mRNA。反转录合成cDNA第一链和第二链,与EcoRI和HindⅢ接头连接。酶切和过柱分级分离后,与λScreen载体连接,经体外包装转染E.coliER1647宿主菌,进行文库容量测定和扩增。以扩增文库的DNA为模板,利用已知基因引物克隆猪IL-2和IL-4的cDNA并进行测序。结果表明,成功构建了猪细胞因子cDNA文库,文库原始库容量为8×105,插入片段在300~2 000 bp,扩增得到特定的IL-2和IL-4基因,说明文库质量高、代表性强,为进一步从文库中筛选未知细胞因子及相关基因提供了有效的工具。     

不同体外培养条件对家兔早期囊胚贴壁行为的影响

采集家兔自然交配后96h的早期囊胚，以低糖DMEM＋150mL／L胎牛血清＋0．1mmol／L非必需氨基酸＋100IU／mL青霉素＋100IU／mL链霉素为基础培养基，比较了不同胚胎处理方法、不同饲养层以及培养液中添加不同成分对兔早期囊胚贴壁和增殖的影响，以完善兔胚胎干细胞的建系方法。结果表明，以胚胎分割法和链霉蛋白酶-E（proteinaseE）处理掉黏蛋白及部分透明带的胚胎容易贴壁和增殖；在小鼠成纤维细胞饲养层和兔胎儿成纤维细胞饲养层上，胚胎的脱带率差别不大，但在小鼠成纤维细胞饲养层上贴壁率明显提高，且贴壁后内细胞团增殖较快；添加胰岛素、白血病抑制因子和伊巯基乙醇均利于抑制ES的分化和促进内细胞团的增殖。     

二花脸仔猪在运输过程中的行为特征和血浆皮质醇水平的变化

本试验采用实时跟踪摄像和颈静脉血管瘘连续采样,观察二花脸断奶仔猪在2h运输过程中的行为表现,并测定血浆皮质醇水平的动态变化。结果表明:运输前期(出发后15min)仔猪的探究行为和异常行为发生次数较多且持续时间较长,运输中期(出发后60~75min)此类行为显著减少(P<0.05),而到运输后期(最后15min)几乎观察不到此类行为;仔猪的躺卧行为在运输前期和中期发生次数较少,持续时间较短,而在运输后期仔猪大部分时间都处于躺卧状态(P<0.05);运输至中期仔猪大部分时间处于站立状态,而到运输后期站立行为显著减少(P<0.05);仔猪蹲坐行为发生的次数和持续时间在运输的前中后无显著差异(P>0.05)。仔猪上车后皮质醇水平迅速上升(P<0.05),出发15min升至峰值,且保持该高水平直到2h运输结束,下车后15min皮质醇水平迅速下降至上车前15min的基础水平。     

Effect of basil polysaccharide on rat model bearing NuTu-19 cells

AIM：To study the effect of basil polysaccharide on the rat model bearing NuTu-19 cells.METHODS：The rat model bearing NuTu-19 cells was treated with basil polysaccharide.The changes of the tumor cell biology and histopathology were observed.Total RNA was isolated from the tumor tissue.Hybridization assay was performed using gene expression microarray.The results were analyzed by photo disposal software-GenePix Pro3.0.RESULTS：Compared with control,the amounts of ascites in the rats treated with basil polysaccharide were decreased (P<0.05) and the scores of abdominal carcinoma metastasis were changed significantly (P<0.01).268 differentially expressed genes were found in the tumor specimen from the rats exposed to basil polysaccharide.CONCLUSION：Basil polysaccharide may be a new anti-tumor drug and it has multi-targets.Gene expression microarray has huge advantage and value in the study of the drug target.     

14-3-3σ negatively regulates Akt and inhibits Rat1-Akt cell xenografts

AIM:The present study was designed to investigate the effect of adenovirus-mediated 14-3-3σ on Rat1-Akt cell xenografts and to explore whether the effect was mediated through negative regulation of Akt. METHODS:The effect of Ad-14-3-3σ on Rat1-Akt cells xenografts was observed in nude mice ex vivo and in vivo. Western blotting was used to detect 14-3-3σ protein,phospho-Akt (Thr 308), phospho-Akt (Ser 473), and phospho-(Ser/Thr) Akt substrate in tumor tissue after transfection of 14-3-3σ gene. RESULTS:The tumor volume was dramatically decreased and its emergence was delayed, regardless of using Ad-14-3-3σ-treated Rat1-Akt cells (ex vivo) or injected Ad-14-3-3σ in vivo, of which the effect of the continuously injected group was the best. Levels of Akt protein, phosphorylated Akt and phosphorylated Akt substrates in tumors obtained from Ad-14-3-3σ-treated mice were markedly less than those in PBS or Ad-β-gal-treated mice. CONCLUSION:14-3-3σ suppressed Akt overexpession in cells of Rat1-Akt xenografts by negatively regulating Akt.     

Pretreatment of hypertonic saline attenuates the hepatic ischemia reperfusion injury induced by neutrophils

AIM: To explore the effect of the pretreatment of hypertonic saline (HTS) in hepatic ischemia reperfusion (I/R) injury.METHODS: The rats were divided into sham group (sham group), ischemia reperfusion group (IR group) and pretreatment of hypertonic saline group (HTS group). Partial hepatic ischemia reperfusion model was used. The rats were sacrificed at the time of 1 h, 3 h, 6 h, 12 h and 24 h after reperfusion in each group, respectively. Blood samples were obtained to examine ALT. The expression of the CD11b/CD18 (Mac-1) on the neutrophils was analyzed by flow cytometry. RT-PCR and Western blotting were used to examine the expression of intercellular adhesion molecule-1 (ICAM-1) in livers and chromatometry was performed to detect the activity of myeloperoxidase (MPO) in livers. The morphology of hepatocytes and the structure of sinusoid were observed by histological examinations. RESULTS: ① HTS pretreatment decreased the level of ALT at the time points of 3 h, 6 h and 12 h after reperfusion (P<0.05). ② Mac-1 expression in HTS group was lower at 6 h and 12 h after reperfusion compared with IR group (P<0.05). ③ MPO activity in HTS group was lower at 6 h, 12 h and 24 h compared with IR group (P<0.05). ④ RT-PCR and Western blotting analysis indicated that the pretreatment of HTS inhibited the expression of ICAM-1 in livers after reperfusion. ⑤ Moderate hepatocyte swelling and few neutrophil infiltration were observed in HTS group.CONCLUSION: Pretreatment with HTS has the effect on hepatic ischemia reperfusion injury by inhibiting the expression of Mac-1 on circulating neutrophils and the expression of ICAM-1 in the liver.