固定化细胞发酵半纤维素水解液产木糖醇的研究

固定在海藻酸钙凝胶中的热带假丝酵母(Candida tropicalis)细胞，可有效地利用玉米芯半纤维素水解液生产木糖醇。在摇瓶条件下，采用分批发酵方式，确立了适宜的发酵工艺参数为：32℃，每升氮源含酵母粉2．2g、胰蛋白胨3．7g，发酵液初始pH值6．0，分段改变摇床转速，其中0～24h为210r／min；24～72h为140r／min，固定化细胞凝胶珠与玉米芯半纤维素水解液体积比为1：4。利用固定化细胞重复进行10批次共30d发酵，木糖醇得率平均为73．7％，达到了理论值的80％。固定化细胞密度高、抗逆性强、发酵性能稳定，水解液未经脱色与离子交换便可转化成木糖醇，显示了良好的应用前景。     

Effect of xeroderma pigmentosum group D gene on proliferation of human umbilical arterial smooth muscle cells induced by Ox-LDL

AIM: To investigate the effects of xeroderma pigmentosum group D (XPD) gene on the proliferation of human umbilical arterial smooth muscle cells (HUASMCs) induced by oxidized low-density lipoprotein (Ox-LDL). METHODS: The recombinant plasmid pEGFP-N2/XPD was transfected into HUASMCs by liposome. The cells were divided into blank control group, pEGFP-N2 group, pEGFP-N2/XPD group, Ox-LDL group, Ox-LDL+pEGFP-N2 group and Ox-LDL+pEGFP-N2/XPD group. The proliferation rate of the cells was detected by MTT and EdU assays. The apoptotic rate and cell cycle distribution were analyzed by flow cytometry. The protein levels of XPD, caspase-3, Bcl-2 and Bax were determined by Western blot. RESULTS: Compared with blank control group, the expression of XPD was increased in pEGFP-N2/XPD group (P<0.05). According to the results of MTT and EdU assays, the cell proliferation in pEGFP-N2/XPD group was reduced compared with blank control group (P<0.05). Compared with Ox-LDL group, the cell proliferation in Ox-LDL+pEGFP-N2/XPD group was significantly inhibited (P<0.05). According to the results of flow cytometry, the cell proportion of S phase decreased and the G₀/G₁-phase cell proportion increased significantly in pEGFP-N2/XPD group and Ox-LDL+pEGFP-N2/XPD group compared with blank control group and Ox-LDL group, repectively (P<0.05). Compared with blank control group and Ox-LDL group, the protein level of Bcl-2 decreased and the protein levels of Bax and cleaved caspase-3 increased in pEGFP-N2/XPD group and Ox-LDL+pEGFP-N2/XPD group, respectively (P<0.05). CONCLUSION: XPD inhibits the proliferation of HUASMCs and promotes their apoptosis, and reduces the promoting effect of Ox-LDL on the proliferation of HUVSMCs. XPD may be the target for treatment of atherosclerosis.     

Screening of lentiviral vectors carrying effective siRNA targeting S1P2 gene

AIM:To screen the lentiviral vector carrying siRNA with higher efficiency of suppressing the sphingosine-1-phosphate receptor 2(S1P2) gene expression in the primarily cultured corpus cavernosum smooth muscle cells of spontaneously hypertensive rats (SHR).METHODS:SHR and SD rats (n=5 each) were used for primarily culturing corpus cavernosum smooth muscle cells.The cells were randomly divided into 6 groups:SHR siRNA-1,SHR siRNA-2,SHR siRNA-3,SHR GFP,SHR control (SHR non-transfection group),and SD control (SD rat control group).Each group had 5 samples with 1.0×10⁵ cells of each sample.At 72 h after transfection (MOI=60) with lentiviral vectors carrying S1P2 siRNA into the SHR corpus cavernosum smooth muscle cells,the expression of GFP was observed under fluorescence microscope.The protein expression of S1P2,ROCK1,ROCK2 and eNOS in the corpus cavernosum smooth muscle cells,and the mRNA expression of S1P2,ROCK1 and ROCK2 were determined by by Western blot and RT-PCR.RESULTS:The transfection efficiency of the corpus cavernosum smooth muscle cells in SHR siRNA-1,SHR siRNA-2,SHR siRNA-3 and SHR GFP groups were>80%.Compared with SHR control group,the mRNA levels and the protein expression of S1P2,ROCK1 and ROCK2 in SHR GFP group showed no remarkable changes,while those in SHR siRNA-1,SHR siRNA-2,SHR siRNA-3 and SD control groups were significantly lower than those in SHR control group (P<0.05).The protein expression of eNOS in SHR siRNA-1,SHR siRNA-2,SHR siRNA-3 and SHR GFP groups were not significantly changed as compared with SHR control group,but that in SD control group was significantly higher than that in SHR control group.CONCLUSION:Three groups of siRNA lentiviral vectors targeting S1P2 inhibit the expression of S1P2 in the corpus cavernosum smooth muscle cells of SHR,and by silencing the S1P2 expression,the expression of ROCK1 and ROCK2 is inhibited.Among them,siRNA-1 has the highest inhibitory efficiency.     

不同提取方法的桑叶提取物对小鼠生长性能、抗氧化和免疫功能的作用研究

本试验旨在通过探究不同桑叶提取段及桑叶粗提取物对小鼠生长性能、抗氧化功能、血清生化指标和炎症细胞因子的影响,筛选出一种较有效的桑叶提取方法。试验使用正己烷、乙酸乙酯、乙醇(85%)和水(60～70℃)制得桑叶提取物,另外使用水煎煮的方法制得桑叶粗提物,分别记为A、B、C、D和E。选取108只5周龄无特定病原体(SPF)级ICR雄性小鼠,随机分为6组,对照组饲喂基础饲粮,试验组在基础饲粮中分别添加桑叶提取物A、B、C、D和E(1 200 mg/kg),每组6个重复,每个重复3只小鼠,预试期3 d,正试期28 d。结果显示：与对照组相比,1)E组小鼠的平均日采食量(ADFI)极显著提高(P<0.01);2) C、D和E组小鼠血清谷胱甘肽过氧化物酶(GSH-Px)活性显著升高(P <0.05),B、D和E组血清总超氧化物歧化酶(T-SOD)活性极显著升高(P<0.01);血清高密度脂蛋白胆固醇(HDL-C)含量显著降低(P<0.05),B和D组血清天门冬氨酸氨基转移酶(AST)活性显著升高(P<0.05),B和E组在血清甘油三酯(TG)含量显著降低(P<0...     

广西蚕区主推家蚕品种在不同区域及不同批次饲养的适应性调查

将广西壮族自治区主推的3对家蚕品种分别在桂西北、桂中和桂南几个具有地域代表性蚕区的不同养蚕批次进行适应性饲养试验,调查不同品种的蚕茧产量和品质,以期通过家蚕品种最好的生产应用布局来提高广大蚕农的经济收益.根据试验结果建议:那坡、环江蚕区每年第1、2、3、10、11批次对饲养家蚕品种的选择顺序为桂蚕2号、两广二号或桂蚕N...     

不同播期夏播小豆产量性能动态指标与光温水效应

为探讨光、温、水等气象因子对夏播小豆产量的影响,达到合理利用气象资源、进一步挖掘小豆增产潜力的目的,以中晚熟小豆品种花小豆(V1)和中早熟品系德红5261(V2)为材料,设T1(6月17日)、T2(6月22日)、T3(6月27日)、T4(7月2日)和T5(7月7日)5个播期,形成不同的光、温、水生长环境,对夏播小豆产量...     

一种新的暖季型草坪草--钝叶草

介绍了一种新的暖季型草坪草———钝叶草,该草种具有耐荫、耐水淹、适应能力强、繁殖快等优点,并且植株低矮,管理要求粗放,是一种值得在南方大力推广的暖季型草坪草。     

猎豹与虎猫杯状病毒的分离及其超变区基因比较研究

用F81细胞从上海某动物园患口腔溃疡的猎豹和虎的唾液病料中分离获得两株杯状病毒，经形态学、理化学、生物学鉴定和病毒核酸超变区基因RT-PCR扩增与序列测定证明两株病毒均为猫杯状病毒(FCV)，分别命名为FCV/cheetah/Shanghai/02/2002与FCV／tiger/Shanghai／03/2002。人工感染猫可引起体温升高，口腔溃疡，食欲下降等症状。两株病毒的超变区序列520bp长片段问的同源性为99.2％，与国内桂林虎分离株(TFCV9710)的同源性为74．0％，与国外分离株的总体同源性为58．1％，说明不同宿主或同种不同个体间杯状病毒分离株超变区核酸差异十分显著，符合猫杯状病毒的分子生物学特征。     

Ⅱ型犬腺病毒自然弱毒YCA18株的实验免疫研究

应用YCA18株第8代细胞培养物（YCA18－8）经口鼻和静脉接种犬腺(CAV)SN本＜1：2的易感犬作安全试验，结果未见任何CAV临床症状与病理解剖学改变；用不同剂量的YCA18－8分组免疫CAV易感犬，经SN抗体测定与用CAV强毒攻击试验，结果SN抗体达1：16以上的犬95％以上可获得免疫保护，最低免疫量为10^4TCID50；应用YCA18－8分别免疫母源抗体达1：4和1：8的两组试验犬，第一次免疫14d后SN抗体均未有升高，追加2－3次免疫后SN抗体才逐渐上升至1：16以上的免疫保护水平；用CAV易感犬和MDCK分别将YCA18连续传8代和20代，结果对犬仍然安全，对犬的免疫原性未见下降，最低免疫量仍为10^4TCID50；与CDV和CPV弱毒作免疫互扰试验，结果与各弱毒的单独免疫结果未见差异；将YCA18－8冰冻保存于一30℃，6个月内TCID50仍不低于10^-4/0.1ml，经加明胶－蔗糖低温真空冻干后，4℃和－30℃保存1年，TCID50均仍维持在10^-4/0.1ml以上，经YCA18－20 3次免疫的试验犬，1年后SN抗体仍在最低抗体保护值1：16以上。     

大肠杆菌氯霉素类耐药基因三重PCR检测试剂盒的研究与应用

本研究研制了大肠杆菌氯霉素类药物耐药基因cat1、flor、cmlA基因三重PCR检测试剂盒.结果显示,该试剂盒具有良好的重复性、灵敏度以及保存期长等特点.应用该试剂盒检测107株猪源鸡源大肠杆菌,cat1、flor、cmlA三种基因的检出率分别是 41.1%、51.4%和36.4%.本试剂盒对氯霉素类药物耐药基因的传播、流行调查以及在临床用药方面具有重要的指导意义.