ObjectiveTo observe the longterm effect of suramin on the inhibition of proliferation of human retinal pigment epithelial (RPE) cells in vitro. MethodsRPE cells grown in 9 pieces of 96well plate (12 wells each plate) were divided into experimental and control group, with 6 wells in each group. The concentration of 0.1 ml RPE cells in each well is 5×104 cells/ml. After the change of the medium, RPE cells were treated with suramin (250 μg/ml) in experimental group while treated with nothing in the control group. The medium of the 2 groups were changed to the normal medium after 4 days. At the 1st, 2nd, and 4thday after the addition of suramin and at the 1st, 2nd, 3rd, 5th, 6th, 7th, 9th , 11th and 13th day after removing suramin, 1 plate was randomly selected to stop culturing, and the proliferation of RPE cells were detected by methyl thiazolyl tetrazolium (MTT) assay. ResultsUnder reversed microscope, RPE cells in control group were fused completely at the 7th day after inoculation. The extracellular space of RPE cells in experimental groups was larger than that in the control group, and remained unfused at the 13th day after inoculation. The inhibitory rate of proliferation of RPE cells at the first day after treated with suramin was 14.85% and increased to the highest 25.79% at the 4th day. The first day after the suramincontaining media was removed, the inhibitory rate decreased to 12.35%, and then raised gradually to over 20% at the 3rd to 5th day. Finally, the rate drop to 14.71%. ConclusionSuramin has the long-term effect on the inhibition of RPE cells induced by serum, especially the inhibitive effect after the remove of suramin, which indicates the specific double-peak inhibition during the whole process.(Chin J Ocul Fundus Dis, 2005,21:25-27)
ObjectiveTo investigate the relationship between topical reactive lymphoid hyperplasia and postoperative recurrence and survival of gastric cancer patients. MethodsThe clinical and pathological data of gastric cancer patients who underwent D2 radical gastrectomy from January 2007 to July 2009 were retrospectively analyzed. Based on the number of reactive lymph nodes, cases were divided in to topical reactive lymphoid hyperplasia group (RLH, n=18) and non-RLH group (n=43) by using a median method. The postoperative disease-free survival (DFS) and overall survival (OS) rates of patients in different groups were compared using Kaplan-Meier method and log-rank test, respectively. ResultsThere were no significant difference between the two groups in age, gender, pathological stage, surgical approach, extent of surgery or methods of postoperative chemotherapy (P > 0.05). The median disease-free survival time was 50 months in RLH group, and the median disease-free survival time was 39 months in non-RLH group. DFS of patients in RLH group was significant higher than non-RLH group (66.7% vs. 34.9%, P=0.048). The median survival time was 53.6 months and 52.3 months, respectively, in RLH group and non-RLH group. No difference was found in OS between the two groups (72.2% vs. 60.5%, P=0.338). ConclusionTopical reactive lymphoid hyperplasia reactive the immunity of gastric cancer patients and contact postoperative DFS rate.
OBJECTIVE: To determine the influence of basic fibroblast growth factor (bFGF) on endothelial cell (EC) proliferation in vitro and its possible mechanisms, and to examine the effect of both TNP-470 and dexamethasone (Dex) on the EC proliferation induced by bFGF. METHODS: Human umbilical vein endothelial cells were cultured and the proliferation of EC was quantified by a colorimetric assay using MTT reagent. The expression of nuclear factor-kappa B (NF-kappa B) and ki-67 was detected with SABC immunohistochemical method. RESULTS: bFGF stimulated the EC proliferation and enhanced the expression of NF-kappa B and ki-67 in nucleus; TNP-470 and Dex suppressed EC proliferation induced by bFGF, and reduced the expression of NF-kappa B and ki-67 in nucleus. CONCLUSION: The above results indicate that the possible mechanisms of EC proliferation stimulated by bFGF come from that bFGF can activate NF-kappa B to promote the synthesis of DNA and EC mitosis. TNP-470 and Dex inhibited EC proliferation stimulated by bFGF by inhibiting NF-kappa B.
Objective To investigate the inhibitive effect of E2F decoy oligodeoxynucleotides (E2F decoy ODNs) on cultured human retinal pigment epithelial (HRPE) cells.Methods E2F decoy ODNs or scramble decoy ODNs at varied concentrations were put into the HRPE cells mediated by lipofectamineTM2000. The proliferative activity of HRPE was detected by methythiazolyl-terazollium assay, and the competitive combinative activity of E2F decoy ODNs and transcription factor E2F was detected by electrophoresis mobility-shift assay. Results The proliferation of HRPE was inhibited markedly by E2F decoy ODNs at the concentration of 0.2 μmol/L (P=0.002) in a dose-dependent manner but not by scrambled decoy. The results of electrophoresis mobility-shift assay showed that the combinative activity of transcription factor E2F was abolished completely by E2F decoy ODNs. Conclusions E2F decoy ODNs may sequence-specifically inhibit the combinative activity of transcripti on factor E2F,and inhibit the proliferation of HRPE cells.(Chin J Ocul Fundus Dis，2004，20：182-185)
Objective To explore the expression characteristics of chaperone interacting protein (CHIP) in normal, scar and chronic ulcer tissues and its relationship with wound healing. Methods Twenty biopsies including scar tissues(n=8), chronic ulcer tissues(n=4) and normal tissues(n=8)were used in this study. The immunohistochemical staining (power visionTMtwo-step histostaining reagent) was used to explore the amount and expression characteristics of such protein.Results The positive expression of CHIP was observed in fibroblasts, endothelial cells and epidermal cells in dermis and epidermis. It was not seen ininflammatory cells. The expression amount of CHIP in scar tissues, chronic ulcer tissues and normal tissues was 89%, 83% and 17% respectively. Conclusion Although the function of CHIP is not fully understood at present, the fact that this protein is expressed only at the mitogenic cells indicates that it may be involved in mitogenic regulation during wound healing.
【摘要】 目的 探讨采用不同方法经尿道前列腺等离子双极电切术（plasmakinetic resection of prostate，PKRP）的方法及疗效。 方法 2008年7月-2009年12月，应用不同方法行PKRP治疗156例前列腺增生。患者年龄59~87岁，平均74岁。病程20 d~18年。前列腺重量22~100 g，平均38 g。采用单纯顺行电切法治疗38例，部分剜除分割切除法治疗76例，完全剜除法治疗42例。 结果 156例手术均获成功，手术时间平均90 min。获得前列腺组织12~87 g，平均35 g。术后留置导尿管平均5.5 d，住院时间平均6.5 d。术后组织病理学诊断为良性前列腺增生152例，前列腺癌4例。拔除尿管后均能自主排尿，部分患者术后有尿道刺激症状；术后1个月内出现尿道外口狭窄3例，经尿道扩张治愈。随访时间1~12个月，平均6个月。短期尿失禁3例，时间分别为1周、1个月及3个月；无长期尿失禁。术后3个月国际前列腺症状评分（IPSS）症状评分平均减少24分，生活质量评分平均减少3分。 结论 PKRP安全、有效、并发症少，可针对患者情况采用不同切割方法，效果更佳。【Abstract】 Objective To explore the effects and methods of transurethral plasmakinetic resection of prostate(PKRP). Methods A total of 156 patients with prostatic hyperplasia were treated with various methods of transurethral PKRP from July 2008 to December 2009. Patient’s age ranged from 59 to 87 years,74 years on average. The disease duration was 20 days to 18 years.Method one:anterograde resection in 38 patients; method two:partition retrograde enucleation in 76 patients; method three:completely retrograde enucleation in 42 patients. Results All of the swgeries were successful. The mean duration of the operation was 90 minutes.The collected prostatic specimens were 12-87 g,35 g on average. The mean catheter remaining dwation was 5.5 days.The mean postoperative hospital stay was 6.5 days. Conclusions PKRP is safe and effective. It is effective with various methods of transurethral plasmakinetic resection of prostate.
Abstract: Objective To investigate the effects of haemopoietic stem cell mobilization on vein graft patency and intimal hyperplasia of anastomosis. Methods Twentyfour New Zealand rabbits were randomly divided into experimental group and control group, 12 rabbits in each group. A double side of carotid arteryvein transplantation model was made in each rabbit. One side of vein graft was digested by 0.25% trypsin for complete endothelial denudation before transplantation. Recombinant human granulocyte colonystimulating factor was given by subcutaneous injection 24 hours after operation, once per day in successive 10 days in experimental group, saline was given in the same way in control group. Bone marrow stem cells mobilization was observed after operation, including karyote counts and mononuclear cell proportion in peripheral blood. The patency rate of vein grafts and the degree of anastomosis intimal hyperplasia were observed too. Results The karyote counts (t=8.406,P=0.000)and mononuclear cell proportion(t=31.267,P=0.000) in peripheral blood of experimental group increased significantly 5 days after operation than those in control group. The vein grafts with intact endothelium had higher patency rate in both groups. In the vein grafts with complete endothelial denudation, the patency rate were obviously lower, but it was higher in experimental group than those in control group (67% vs. 30%). In the end of experiment, the pulsatility index of the vein grafts anastomosis with complete endothelial denudation was lower in experimental group than that in control group(t=2.958,P=0.009). Pathological examination showed that various degrees of intimal hyperplasia in all anastomoses of vein grafts were observed 4 weeks after operation. The degree of anastomosis intimal hyperplasia was more severe in vein grafts with complete endothelial denudation. Compared with control group, re-endothelization occurred completely in vein grafts with complete endothelial denudation of experimental group and the degree of anastomosis intimal hyperplasia was relatively lower (Plt;0.05). Conclusion Haemopoietic stem cell mobilization can provide protective effects on vein grafts by accelerating reendothelization which might increase vein grafts patency rate in the near future after operation and reduce anastomosis restenosis caused by intimal hyperplasia.
To investigate the inhibitory effect of Col I A1 antisense ol igodeoxyneucleotide (ASODN) transfection mediated by cationic l iposome on Col I A1 expression in human hypertrophic scar fibroblasts. Methods Scar tissue was obtained from volunteer donor. Human hypertrophic scar fibroblasts were cultured by tissue block method. The cells at passage 4 were seeded in a 6 well cell culture plate at 32.25 × 104 cells/well, and then divided into 4 groups: group A, l iposomeand Col I A1 ASODN; group B, Col I A1 ASODN; group C, l iposome; group D, blank control. At 8 hours, 1, 2, 3 and 4 days after transfection, total RNA of the cells were extracted, the expression level of Col I A1 mRNA was detected by RT-PCR, the Col I A1 protein in ECM was extracted by pepsin-digestion method, its concentration was detected by ELISA method. Results Agarose gel electrophoresis detection of ampl ified products showed clear bands without occurrence of indistinct band, obvious primer dimmer and tailing phenomenon. Relative expression level of Col I A1 mRNA: at 8 hours after transfection, group A was less than groups B, C and D (P lt; 0.05), and groups B and C were less than group D (P lt; 0.05), and no significant difference was evident between group B and group C (Pgt; 0.05); at 1 day after transfection, groups A and B were less than groups C and D (P lt; 0.05), and there was no significant difference between group A and group B, and between group C and group D (P gt; 0.05 ); at 2 days after transfection, there were significant differences among four groups (P lt; 0.05); at 3 and 4 days after transfection, group A was less than groups B, C and D (P lt; 0.05), group B was less than groups C and D (P lt; 0.05), and no significant difference was evident between group C and group D (P gt; 0.05). Concentration of Col I protein: at 8 hours after transfection, group A was less than groups B, C and D (P lt; 0.05), groups B and C were less than group D (P lt; 0.05), and no significant difference was evident between group B and group C (P gt; 0.05); at 1 day after transfection, significant differences were evident among four groups (P lt; 0.05); at 2, 3 and 4 days after tranfection, groups A and B were less than groups C and D (P lt; 0.05), and no significant difference was evident between group A and group B (P gt; 0.05). Conclusion Col I A1 ASODN can inhibit mRNA and protein expression level of Col I A1. Cationic l iposome, as the carrier, can enhance the inhibition by facil itating the entry of ASODN into cells and introducing ASODN into cell nucleus.