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141.
CHEN Ya-hong ZHAO Ming-wu FU Min-gui YAO Wan-zhen WANG Xiao-hong WANG Jian-li TANG Chao-shu 《园艺学报》2002,18(2):157-160
AIM:To investigate the role of calcineurin (CaN) in airway remodeling in guinea pig model of asthma.METHODS:Male guinea pigs were randomly divided into three groups: control, asthma group and CsA group. The following parameters were measured: 1. The protein content, cell count and differential count of BALF; 2. The amount of [3H]-TdR incorporation into central airway smooth muscle; 3. The mean thickness of airway wall and airway smooth muscle of small airwaysl; 4.CaN activity of trachea and lung tissue.RESULTS:1. The protein content, cell count and eosinophil of BALF in CsA group were 46%, 51% and 60% lower than those in asthma group, respectively (P<0.01); 2. [3H]-TdR incorporation in CsA group was 22% lower than that in asthma group (P<0.05);3. The mean thickness of airway wall and airway smooth muscle were 34% and 37% less in CsA group than those in asthma group, respectively (P<0.01); 4. CaN activity of lung tissue and trachea were 52% and 44% lower in CsA group than those in asthma group, respectively (P<0.01).CONCLUSION:CsA reduced airway remodeling in guinea pig model of asthma, indicating the role of CaN in the airway remodeling. 相似文献
142.
AIM: To prepare gfp-bcl-XL-contained recombinant adenovirus(rAd-gfp-bcl-XL).METHODS: Bcl-XL gene was amplified from pEGFP-C3-bcl-XL, subcloned into shuttle plasmid and formed transfer plasmid of pAdTrack-CMV-bcl-XL. Then pAdTrack-CMV-bcl-XL 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-XL and pAdEasy-1 into BJ5183. Recombinant adenovirus particle were produced and further amplified after the transfection of pAdEasy-1-gfp-bcl-XL into 293 cells. PCR test indicated that the recombinant Ad contained bcl-XL gene. The titer of the purified rAd-gfp-bcl-XL was 6.5×1012 PFU/L. CONCLUSIONS: The homologous recombination in bacteria is a convenient and high efficient method to prepare rAd-gfp-bcl-XL. This affords a good gene transfer vector for the gene therapy in human’s diseases. 相似文献
143.
AIM and METHODS: Total RNA was extracted from 6th rat subcultured pulmonary artery smooth muscle cells(PASMC) exposed to continual chronic hypoxia or normoxia and the effects of chronic hypoxia on the changes of Kv1.3,Kv2.1,Kv3.1 mRNA in cultured PASMC induced by acute hypoxia were studied by semiquantitative RT-PCR in vitro. RESULTS:①Kv1.3,Kv2.1,Kv3.1 genes were found to be expressed in PASMC of rats exposed either to hypoxia or normxia.②The expression of Kv2.1 and Kv3.1 in 6th subcultured of PASMC in normaxia group could be upregulated by exposure to acute hypoxia,the levels of Kv2.1 and Kv3.1 mRNA were significantly increased from 0.646±0.092, 0.782±0.104 to 1.059±0.134, 0.985±0.116,respectively (P<0.01,n=5). ③PASMC cultured continuously in chronic hypoxia for 6 subcultures and then exposed to normoxia for 12 h,thereafter the expression of Kv2.1 and Kv3.1 were downregulated by acute hypoxia for 6 hours.The level of Kv2.1 mRNA was significantly decreased from 1.008±0.117 to 0.649±0.097 (P<0.01,n=5). CONCLUSION:Kv2.1,Kv3.1 genes might be oxygen sensitive genes.Chronic hypoxia might change the response of these Kv genes of PASMC to acute hypoxia and down-regulate its expression,which might probably decrease the role of Kv in HPV. 相似文献
144.
AIM: To examine the expression of human endostatin in E.coli, produce its fusion protein antibody and observe its biological activity. METHODS: Endostatin gene was amplified by polymerase chain reaction,recombined with plasmid vector pGEX-2T and induced expression with IPTG.The protein activity was tested by endothelial cell proliferation inhibitory assay.Inclusion body crudely purified was used to generate polyclonal antibody to detect its expression at mouse's liver and kidney etc. RESULTS: The protein expressed was 20kD after digestion by thrombin,it appeared the anti-angiogenesis activity and Western blotting indicated the expression of endostatin in liver and kidney of mouse. CONCLUSION: The successful expression of human endostatin and the preparation of polycolonal antibody indicated its potential application in anti-angiogenesis therapy and diagnosis tumors. 相似文献
145.
REN Yu-sheng WU Zong-gui CUI Fang JIA Guo-liang YU Shi-qing TANG Chao-wu LI Bo 《园艺学报》2002,18(11):1377-1380
AIM: To investigate the effects of platelet-derived growth factor on DNA and collagen protein synthesis in human vascular fibroblasts. METHODS: In the present experiment, the human vascular fibroblasts were cultured and effects of platelet-derived growth factor-BB on DNA and collagen protein synthesis in human vascular fibroblasts were observed by using [3H]-TdR incorporation and [3H]-proline incorporation in vitro. RESULTS: Platelet-derived growth factor-BB significantly promoted NDA synthesis and collagen protein synthesis of quiescent human vascular fibroblasts, with a maximal response at a concentration of 30μg·L-1at 24 h and 36 h, respectively. CONCLUSION: Platelet-derived growth factor-BB promotes DNA and collagen protein synthesis in cultured human vascular fibroblasts. 相似文献
146.
TANG Yi-da YANG Yue-jin RUAN Ying-mao LI Yong-li ZHOU Yan-wen TIAN Yi GAO Run-lin CHEN Ji-lin CHEN Zai-jia 《园艺学报》2002,18(9):1115-1118
AIM:To compare the effects of carvedilol, cilazapril and their combination on left ventricular remodeling(LVRM) after acute myocardial infarction(AMI) in rats. METHODS: Twenty-four hours after AMI operation, 100 surviving rats were randomly assigned to: ①AMI control(n= 25), ②AMI+carvedilol(1 mg·kg-1 ·d-1, n= 25)(C1), ③AMI+cilazapril(1 mg·kg-1 ·d-1, n= 25)(Z1), and ④ AMI+combination(n= 25) groups. Sham-operated group(n= 17) were selected randomly. After 4 weeks of therapy with the drugs gastric gavage, hemodynamic and pathological studies were performed. RESULTS: There were no significant differences in MI size among the four AMI groups(all P> 0.05) Left ventricular(LV) end diastolic pressure(LVEDP), volume(LVV), weight(LVW) and septal thickness(STh) were all higher and left ventricular pressure maximal rate of rise and fall(±d p /d t) were lower(all P< 0.01) in AMI group than sham-operated group. The LVEDP, LVV, LVW and STh were all lower and ±dp /dt were higher in Z1, C1, and combination groups than those in AMI group(P< 0.05, P< 0.01), with LVEDP and STh were more lower in the combination group than in the two monotherapy group(P< 0.05, P< 0.01), but there were no significant differences in other variables among the three therapy groups. CONCLUSION: Carvedilol, cilazapril and their combination all can prevent from LVRM after AMI in rats, improve hemodynamics and LV function, with the combination superior. 相似文献
147.
ZHANG Yu-xia YU Lun-yin LIU Ming-qiu ZHANG Zheng-bin TANG Zhi-jiao XIA Dong WANG Ming 《园艺学报》2002,18(1):32-35
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. 相似文献
148.
149.
肥城桃组培苗诱导、基因转化及其增殖 总被引:11,自引:0,他引:11
红里、白里是肥城桃的两个优良品种,但不耐贮藏。为试图利用基因工程解决这一问题,先通过组培将其胚、胚乳、子叶的愈伤组织分别诱导再生植株和芽剥离出茎尖培育成苗。再将肥城桃反义PG基因,通过农杆菌介导转化组培苗,转化苗在含卡那霉素培养基上筛选,获得抗卡那霉素的转基因苗。用研制的增殖培养基培养4~7周,一个苗可生长出14~18分枝的苗,解决了转基因苗获得率低的问题,也满足了对转基因株系进行检测和从组培室到温棚到田间果园过渡栽培和芽接的需求,提高了繁育速率。 相似文献
150.
实验室条件下系统研究了磷化氢(PH3)对储物害虫嗜卷书虱Liposcelis bostrychophila卵、各龄若虫和成虫的致死作用,并选用PH3间歇熏蒸以及PH3与气调交替处理等措施对嗜卷书虱进行处理,比较了不同处理措施对嗜卷书虱种群的控制效果。结果表明,PH3熏蒸处理对嗜卷书虱各虫态有不同的致死效果。对卵而言,24、72和120 h熏蒸处理的LC50分别为0.137、0.045和0.035 mg/L;而24 h熏蒸处理对若虫的LC50在4.285~7.364 μg/L之间,对成虫的LC50为20.404 μg/L;采用25 μg/L的PH3进行24 h熏蒸处理,间隔10 d后再分别进行第2次和第3次熏蒸处理,可以完全控制嗜卷书虱的发生。采用PH3 (12 μg/L)和气调(体积比例为35% CO2,1% O2,64% N2)交替处理能够延缓嗜卷书虱种群抗性的发展,交替处理3~5次可以完全控制嗜卷书虱的发生。 相似文献