ObjectiveTo investigate the proliferation and apoptosis effects of adenovirus-mediated interleukin-24 (Ad-IL-24) gene on Karpas299 cells in vitro. MethodsThe Karpas299 cells were divided into blank control group, Ad-IL-24 group, and the adenovirus which carrying green fluorescent protein gene group (Ad-GFP group). Karpas299 cells of Ad-IL-24 group were infected by adding 200.0 μL Ad-IL-24, Karpas299 cells of Ad-GFP group were infected by adding 200.0 μL Ad-GFP, but Karpas299 cells of blank control group were treated by adding 200.0 μL PBS. Cells' proliferation inhibition rates of 3 groups were detected by cell counting kit (CCK-8) method at 12, 24, and 48 hours after treatment, respectively, and the cells' apoptosis rates of 3 groups were detected by flow cytometry at 48 hours after treatment. ResultsAd-IL-24 can suppress the growth of Karpas299 cells, and the inhibition rate increased over time. Compared with Ad-GFP group at the same time, the cell' proliferation inhibition rate of Ad-IL-24 group was higher at 12, 24, and 48 hours after treatment (P<0.05). In addition, the cells' apoptosis rate of Ad-IL-24 group was higher than those of Ad-GFP group and blank control group at 48 hours after treatment (P<0.05). ConclusionAd-IL-24 can suppress the growth of Karpas299 cells and induce the apoptosis of it.
OBJECTIVE To probe the possibility of direct transfer of exogenous gene into peripheral nerve and its following expression in vivo. METHODS The PCMV beta plasmid containing cytomegalovirus (CMV) promoter and Escherichia Coli (E. Coli), beta-Galactosidease (beta-Gal) structural gene (lacZ gene) was constructed and injected into the rabbit sciatic nerve. The control group was injected PBS solution. The injected nerves were sampled and tested by beta-Gal enzyme activity assay of the 5-bromo-4-chloro-3-indolyl-beta-D-galactoside and beta-Gal histochemical stain. RESULTS In the control group, no beta-Gal enzyme activity was detected in the different stages after operation, and beta-Gal histochemical stains showed positive. In the experimental group, enzyme activity could be detected from 2 days to 30 days after operation, and the histochemical stains showed negative. CONCLUSION The exogenous gene can be transferred into peripheral nerve and expressed with bioactivity, thus the gene therapy to accelerate the recovery of nerve is practical.
Objective To evaluate the inhibited effects of small interfering RNA targeting Rac1 (Rac1-siRNA) on rat retinal neovascularization in retinae. Methods Retinal vein occlusion was induced by retinal photodynamic medthod in 25 Sprague-Dawley rats. Rac1-siRNA vector DNA was injected into the vitrous of one eye of those rats (gene intervention group), and empty vector DNA was injected into the fellow eye (blank control group). Rac1-siRNA vector was injected in other 25 SD rats without retinal vein occlusion (blank intervention group). Two weeks after injection, fluorescein isothiocyanate (FITC)-dextran was perfused into the hearts of all the rats, and the retinal wholemount was made to observe the neovascularization. The numbers of endothelial cells which break through the internal limiting membrane were counted after hematoxylin-eosin staining. Results A massive of neovascularization and FITC leakage were found in blank control group. Small part of neovascularization and a little FITC leakage were observed in the gene intervention group. Retinal vessels were normal in blank intervention group. Compared with blank contrast group and blank intervention group, the difference of the mean numbers of endothelial cells which broke through the internal limiting membrane in the gene intervention group was significant（t=? P=0.000??lt;0.05）. Conclusion Rac1-siRNA can inhibit retinal neovascularization induced by retinal vein occlusion in rats.
Objective To review the research and del ivery methods of growth factors in tendon injuries, and to point out the problems at present as well as to predict the trend of development in this field. Methods Domestic and international l iterature concerning growth factors to enhance tendon and l igament heal ing in recent years was extensively reviewed and thoroughly analyzed. Results Cell growth factor could promote tendon heal ing, improve the mechanical properties as well as reduce the adhesion postoperatively. The use of transgenic technology mediating cell factors to promote tendon repair shows its advantages in many ways. Conclusion The growth factors play a vital role in tendon heal ing. Reasonable treatment of growth factors through direct appl ication or gene transfer techniques is of great value for the heal ing process.
摘要:目的:研究高血压病患者脂蛋白脂肪酶(liportein lipase, LPL)S447X基因多态性与认知功能之间的关系。方法: 对2008年1月至2008年11月在四川大学华西医院医院门诊就诊的原发性高血压患者190例,收集一般资料,采用国际通用的简易智力状况量表测验认知功能,计算认知评分,用聚合酶链反应限制性片段长度多态性(PCRRFLP)技术测定LPL S447X基因多态性。同时测定胆固醇、甘油三酯、空腹血糖、空腹胰岛素及餐后2h血糖、餐后2h胰岛素水平。结果: 高血压病患者认知功能正常组和认知功能障碍组组间LPLS447X基因的基因型和基因频率差异均无统计学意义(Pgt;0.05), SS和SX频率分别为92.6%、7.4%,S和X等位基因频率分别为96.3%和3.7%。结论: LPLS447X 基因多态性可能与高血压认知功能障碍无明显相关性。Abstract: Objective:To study the relationship between liportein lipase(LPL) S447X polymorphism and cognitive function in patients with primary hypertension. Methods:One hundred and ninety hypertensive patients from January 2008 to November 2008 in West China Hospital of Si Chuan University. We collected the general data and applied the Mini Mental State Examination to test the cognitive function and computed score. PCRRELP method was used to analyze the LPL S447X gene polymorphism. Total cholesterol、triglyeride、fasting plasma glucose and postprandial blood sugar、fasting insulin and postprandial plasma insulin were collected. Results:In primary hypertensive patients, both of the genotype frequency and the allele frequency of the LPL S447X polymorphism were not different between the cognitive normal group and the cognitive impaired group (Pgt;0.05). SS genotype was present in 0926 of the population, SX genotype was present in 0.074 of the population. allele frequencies were 0.963 for S allele and 0.037 for X allele. Conclusion:This results suggest S447X polymorphism in LPL with primary hypertension may not be associated with cognitive impairment. And age and postprandial plasma insulin level are the risk factors of hypertensive cognitive impairment.
Objective To review the recent studies on the suppressing function of breast cancer metastasis suppressor 1 (BRMS1) in breast cancer metastasis. Methods The recent literatures on the mechanisms of BRMS1 in the breast cancer that were published in and abroad were reviewed and summarized. Results BRMS1, similar to the other anti-metastasis genes, only suppresses the metastasis of breast cancer cells but has nothing to do with the growth of tumor. BRMS1 could suppress metastasis of tumor cells by reestablishing both the homospecific and the heterospecific gap junctional intercellular comminications (GJIC) and by altering the expressions of relevant metastasis genes in the breast cancer. Conclusion Further studies on BRMS1 may be helpful to understand the metastasis of breast cancer, which may provide a new way for the diagnosis and treatment of breast cancer.
Objective To investigate the mRNA expression of ciliary neurotrophic factor on the retina during injury and repair of optic nerves in rats. Methods Thirty-five healthy SD rats were randomly divided into 3 groups: 5 in the control group, 15 in the simply transected optic nerve group and 15 in the optic nerve-sciatic nerve anastomosis group. The simply transected and optic nerve-sciatic nerve anastomosed models were set up, and the retinal tissues of all of the rats were taken out after 3, 7 and 14 days, respectively; and the mRNA expression of CNTF in the 3 groups were observed by semiquantitative reversal transcription-polymerase chain reaction method. Results A minimum expression of CNTF mRNA was found in the retinae of the control group, and the increased rates of expression were found in the retinae of the simple transection of optic nerve group with the increase rate of 100%, 594%, and 485% on the 3rd, 7th, and 14th day respectively after the operation, while in optic nerve-sciatic nerve anastomosis group, the increase rates were found to be 258%, 752% and 515% on the 3rd, 7th, and 14th day respectively after the operation. Conclusion Retinal neurons can respond to axonal reaction of retinal ganglion cells by up-regulate endogenous CNTF after the injury of the optic nerves, which may provide a theoretic base for the application of the exogenous CNTF. (Chin J Ocul Fundus Dis，2004，20：355-357)
ObjectiveTo observe the mutation and expression of Nkx2.5 in congenital heart disease patients with diminutive pulmonary blood. We preliminarily explored the association between Nkx2.5 gene and pathogenesis of congenital heart disease patients with diminutive pulmonary blood. MethodsFifty six patients of congenital heart disease with diminutive pulmonary blood in the first affiliated hospital of Bengbu medical college and Anhui province children, s hospital between May 2012 and May 2014 were as an experimental group. Sixty three patients of ventricular septal defect were as a control group. In the trial group, there were 30 males and 26 females averagely aged 5.82± 4.23 years ranking from 6 months to 14 years. In the control group, there were 36 males and 27 females averagely aged 6.93± 4.56 years ranking from 6 months to 14 years. Before operation, peripheral venous blood of all the patients were collected. We used polymerase chain reaction combined with DNA sequencing technology to detect Nkx2.5 gene exon sequence and to analyze the association between Nkx2.5 gene mutation and congenital heart disease with diminutive pulmonary blood. And we got some hypertrophic myocardial tissue from right ventricular outflow tract in the operation, whose size was 0.5× 0.5× 0.5 cubic centimeter. And we extracted myocardial tissue RNA. The expression changes of Nkx2.5 gene mRNA were detected by real-time fluorescence quantitative polymerase chain reaction technique. ResultsThere was no mutations tested out in the peripheral venous blood in both two groups. The expression of mRNA in Nkx2.5 gene of the trial group was lower than that in the control group with a statistical difference. ConclusionNkx2.5 gene mutation may be associated with multiple factors. The occurrence of congenital heart disease with diminutive pulmonary blood may be related with a decline of Nkx2.5 gene expression in the myocardial tissue.