Preparation of gfp-bcl-XL-contained recombinant adenovirus vector by the homologous recombination in bacteria

AIM: To prepare gfp-bcl-X_L-contained recombinant adenovirus(rAd-gfp-bcl-X_L).METHODS: Bcl-X_L gene was amplified from pEGFP-C₃-bcl-X_L, subcloned into shuttle plasmid and formed transfer plasmid of pAdTrack-CMV-bcl-X_L. Then pAdTrack-CMV-bcl-X_L was linealinzed with PmeI and co-transformed into BJ5183 bacteria with adenovirus genomic plasmid of pAdEasy-1. The identified recombinant adenovirus plasmid was digested with PacI and transfected into 293 cells to package recombinant adenovirus particles. The target gene was detected by PCR.RESULTS: There were about 35% positive recombinant bacterial clones after the co-transformation of pAdTrack-CMV-bcl-X_L and pAdEasy-1 into BJ5183. Recombinant adenovirus particle were produced and further amplified after the transfection of pAdEasy-1-gfp-bcl-X_L into 293 cells. PCR test indicated that the recombinant Ad contained bcl-X_L gene. The titer of the purified rAd-gfp-bcl-X_L was 6.5×10¹² PFU/L. CONCLUSIONS: The homologous recombination in bacteria is a convenient and high efficient method to prepare rAd-gfp-bcl-X_L. This affords a good gene transfer vector for the gene therapy in human’s diseases.     

Protein expression of cyclin D2 and p16 in proliferation and differentiation of cultured cardiac myocytes

AIM:To investigate the protein expression of cyclin D₂ and p16 in proliferation and differentiation of cultured cardiac myocytes.METHODS:One-day-old Sparague-Dawley rats were used. Cardiac myocytes(CM) were collected by a trypsin-dispersal method and cultured. Cell growth line and fluorescence activated cell sorting (FACS) were used to investigate the proliferation of CM. Ultra-thin sections were made to observe the ultrastructure of CM under transmission electron microscope. The expression of cyclin D₂ and p16 in CM were measured using immunocytochemistry and image analysis.RESULTS:①Results of cell growth line and FACS analysis showed that cultured CM could proliferate in the first 3 cultured days, but the ability decreased quickly, concomitant with differentiation. CM was obseved quiescent in cell cycle three days later. The ultrastructure of CM showed the large amount of myofilaments and mitochondrion. ②The protein expression of cyclin D₂ in 3,4,5 day CM group was 0.89 times(P＜0.05),0.80 times (P＜0.05) and 0.56 times (P＜0.01) of that in 1 day group, respectively. The expression of p16 in CM was increased during the culture process, 2,3,4,5 day group were 1.63 times, 1.72 times, 1.99 times and 2.84 times (P＜0.01) of that in 1 day group, respectively.CONCLUSION:Cultured neonatal rat cardiac myocytes could proliferate during the first 3 days after incubation, but the ability of proliferation decreased, from the fourth day, concomitant with differentiation. Cyclin D₂ and p16 play the key roles in CM postnatal development. Downregulation of cyclin D₂ and upregulation of p16 may induce CM differentiation.     

两种高水溶性β -环糊精衍生物对甲基对硫磷的增溶和光催化降解作用

研究了两种β-环糊精的衍生物甲基-β-环糊精(MCD)和羟丙基-β-环糊精(HPCD)对甲基对硫磷的增溶作用和对紫外光降解的影响。结果表明:MCD和HPCD能增强甲基对硫磷的水溶性,在25℃下,20 g/L的MCD和HPCD溶液中,甲基对硫磷溶解度比在纯水中分别提高了21.91和17.92倍;另外,3 g/L、6 g/L MCD和HPCD分别处理的甲基对硫磷,其光降解速率分别加快了4.87~6.85倍。增溶作用和光敏效应主要是由于MCD和HPCD与甲基对硫磷形成包合物引起的。     

白菜不同品种对硝酸盐吸收积累差异原因初探

 用两组积累硝酸盐不同的白菜品种作试材，测定其吸收硝酸盐的速率、动力学常数Km与V一、硝酸还原代谢库内硝酸根浓度(MPS)和硝酸还原酶活性(NRA)，结果表明，不同白菜品种吸收硝酸根的速率、Km与V 、MPS、NRA存在较大的差异；硝酸盐积累的差异主要是由于吸收硝酸根的速率不同，与硝酸根在其体内的运转、代谢并无显著直接相关。     

不同温度下转换寄主对桃蚜生态学特征的影响

在5种温度、4组寄主与蚜源组合处理条件下研究了温度对转换寄主后的桃蚜的影响。同一寄主间移植的桃蚜在有效积温、发育起点温度、生殖率、死亡率、寿命以及抗逆性上与不同寄主间移植的桃蚜有明显的差异;寄主对桃蚜生物学、生态学特征有明显的影响,表明长期生活在一种寄主上的桃蚜获得了一定的专化性。若将其移植到另一种寄主上,则会表现出种种不适反应,但这种反应还未进化至型或寄主生殖隔离。     

国产克洛素隆尿药排泄数据分析

应用Kromasil-C18色谱柱(250 mm×4.6 mm,5 μm),WatersTM480型可调波长紫外检测器,0.01M磷酸钾(pH=7):乙腈(3:1)为流动相,检测波长265 nm,含量测定采用标准曲线法,建立了RP-HPLC法检测绵羊尿中克洛素隆含量的方法.方法有效性评价结果表明,尿药含量在0.01～5.0μg/ml及5.0～30.0μg/ml范围呈良好线形关系(r=0.9993、0.9995),方法平均回收率99.32%,日内、日间变异系数分别为3.91%、6.28%.尿药最低检测限0.005μg/ml.试验绵羊以7 mg/kg单剂量经静脉、肌肉及口服三种途径给药后,尿中药物浓度分析表明,克洛素隆经体内处理后主要经肾脏排泄,给药96 h内,静注经尿排泄原药为81.76%,肌注为64.03%,口服为48.50%.     

富铁酵母对家兔红细胞免疫功能的影响

为探讨富铁酵母对家兔红细胞免疫功能的影响,将32只家免随机分成富铁酵母高、低剂量组,FeSO4组及空白对照组,分别灌服富铁酵母(Fe元素8 mg/kg、4 mg/kg)、FeSO4(Fe元素4 mg/kg)及空白生理盐水;连续5 d后,检测红细胞C3g受体花环率和复合物(IC)花环率.结果:富铁酵母较FeSO4及空白组能显著提高红细胞C3h受体花环和红细胞复合物(IC)花环的百分率,但高、低剂量间无显著性差异.结论:富铁酵母能显著提高红细胞免疫功能.     

多花黑麦草品种比较试验

通过对现行生产中常用的4个多花黑麦草品种在昆明地区的物候期、株高、草产量、生长率、净同化率等指标的规定表明,在生长初期,特高的生产性能比其余3个品种都好,但生长一段时间后,邦德的生产性能明显超过特高(P<0.05),抗锈王略高于特高(P>0.05),旺饲的生产性能最差(P<0.05)。因此,邦德是一个在生产中值得推广应用的品种。     

温度对杨树菇栽培性状影响的统计学分析

分析了温度环境因子对杨树菇生长发育的影响 ,研究结果表明 :菌丝体营养生长以 2 5～ 2 7℃为最适。原基分化温度 11～ 16℃ ,子实体发育以 13～ 2 6℃最适 ,不同菌株的最适温度存在差异。通过对日平均气温与杨树菇生产周期和产量、品质的关系试验 ,温度与病害发生关系试验 ,用统计分析方法得出了杨树菇各生理阶段的最适温度范围 ,为杨树菇的优质高产栽培提供了温度环境因子的基础数据和调控依据。     

桃树根癌病防治技术探讨

桃树根癌病是公认最难防治的土传细菌病害之一。在对该病进行较系统研究的基础上,采用多种抗菌药剂、多种方法进行了室内、外药敏测定与防治试验,显示出异菌氰、次氯酸钠、乙蒜素、妥布霉素、环丙沙星、庆大霉素及其混用对防治桃树根癌病菌有明显防效。