Objective To establish a method for primary culture of iris pigment epithelial cells(IPE). MethodsEnzyme-Assisted microdissection was used to isolate and cultivate the IPE cells.An identification was made with microscopic and immunohistochemical observations.Results IPE were successfully sultured and showed on differences with RPE in primary culture and subculture.ConclusionEnzyme-Assisted microdissection is a reliable and quick method for the isolation of IPE.
Objective To observe the characteristics of fundus autofluorescence （AF） distribution at the posterior pole in normal subjects. Methods Seventy-nine normal subjects （156 eyes）were studied. Confocal scanning laser ophthalmoscope (cSLO) HRA2 was used to obtain the AF average image at the posterior pole. The distance was calibrated by Digimizer image analysis system. With umbo as the center, the macula was divided into foveola, fovea, parafovea and perifovea areas which with the radius 175, 750, 1250 and 2750 mu;m respectively. Each area was further divided into inferior, superior, temporal and nasal quadrants by two radial lines angle of 90deg;, except for foveola. The AF intensity in four quadrants of different macular regions and optic disc were measured. The AF intensity in vertical and horizontal direction of umbo was also measured. Then the effects of age, eyes, and gender on AF intensity in four quadrants of different macular regions were analyzed. Results There were statistically significant differences in AF intensity among optic disc and four quadrants of macular regions （F=528.648, P＜0.05）. AF distribution was V-type in vertical direction and M-type in horizontal direction. There were statistically significant differences between age groups in foveola, inferior parafovea, temporal parafovea, inferior perifovea, superior perifovea and temporal perifovea （P＜0.01）. There were no statistically significant differences between the two eyes （P＞0.05）. Between genders group, there were statistically significant differences in foveola, superior fovea, inferior fovea, nasal fovea and temporal perifovea （P＜0.05）; no statistically significant differences in the other quadrants （P＞0.05）. Conclusions The distribution of AF intensity is inhomogeneous in macular regions and four quadrants of each region in normal subjects. AF intensity increases with aging. AF distribution is symmetrical in both eyes. There is probably no correlation between gender and AF intensity distribution.
【摘要】 目的 观察不同种培养基中重组人色素上皮衍生因子(rPEDF)融合蛋白的表达。 方法 将前期研究已构建的pET28aPEDF原核表达重组体转化E.coli BL21大肠杆菌表达宿主菌,酶切鉴定阳性菌落后,分别在M9和LB培养基中用异丙基βD硫代半乳糖(IPTG,IsopropylbetaDthiogalactoside)诱导表达,SDSPAGE电泳检测表达的PEDF蛋白, 美国ImagePro Plus 分析系统进行蛋白定量分析。结果 LB和M9培养基中均获得相对分子质量约54×103的rPEDF融合蛋白。但LB培养基获得的是rPEDF融合蛋白的包涵体,目的蛋白占总蛋白含量为21046%,M9培养基获得的是可溶性的rPEDF的融合蛋白,目的蛋白占总蛋白含量的1231%。结论 不同种培养基中均有rPEDF 融合蛋白的表达。【Abstract】 Objective To observe the express of recombinant pigment epithelial derivative facto (rPEDF) in the different medium. Methods The pET28aPEDF was transformed into E.coli BL21. After the colonies were positive identification which were induced by IsopropylbetaDthiogalactoside in medium M9 and LB. The PEDF protein were detected by SDSPAGE and analyzed by American ImagePro Plus system. Results LB and M9 medium obtained the relative molecular mass about 54×103 rPEDF fusion protein. But LB medium obtained the inclusion bodys of rPEDF fusion protein，the purpose protein account for 21.046%；LB medium obtained the soluble rPEDF fusion protein，the purpose protein account for 12.31%. Conclusion The rPEDF protein was expressed in the different medium.
Systemic lupus erythematosus is an autoimmune disease involving multiple organs of the body. Lupus nephritis is one of the most serious organ manifestations of systemic lupus erythematosus. Vimentin, a member of the intermediate filament protein family, is involved in the pathogenesis of many autoimmune diseases, including lupus nephritis. More and more studies have shown that vimentin plays an important role in the pathogenesis of lupus nephritis, and has an important influence on the disease development, treatment and prognosis of lupus nephritis. This review focuses on the structure, function and post-translational modification of vimentin, the relationship between vimentin and the pathogenesis of lupus nephritis, and the significance of vimentin expression levels in renal tissues, serum and urine, in order to provide theoretical basis for future mechanism research and clinical application.
摘要:目的: 探讨联合LCT和高危型HPV检测对CIN宫颈治疗后的随访意义。 方法 :对200例LCT异常,高危型HPV阳性,阴道镜活检证实为CIN1~3的患者行LEEP治疗或宫颈冷刀锥切,治疗后进行严格随访,包括LCT和高危型HPV检测,阳性病例行组织学检查。 结果 :(1)所有病例经治疗后均无病变残留,其治愈率为100%。(2)从治疗后3个月起,CIN1组高危型HPV转阴率为100%。在随访的第3个月和6个月,CIN2~3组高危型HPV转阴率分别为7317%和9085%,显著低于CIN1组,差异有统计学意义(〖WTBX〗P ＜005)。(3)从随访12个月起,一直有2例病例持续HPV阳性,均为CIN3患者,但LCT和阴道镜检查未发现细胞学异常,继续随访。 结论 :CIN治疗后高危型HPV的转阴时间及转阴率与CIN的级别有关;高危型HPV持续阳性,但LCT和阴道镜检查无异常者可继续严格随访;LCT联合高危型HPV检测是CIN治疗后临床追踪随访的有效手段。Abstract: Objective: To investigate the Significance of LCT joint highrisk HPV testing for followup after CIN treatment. Methods : 200 cases that highrisk HPV infection were tested by realtime PCR and CIN1~3 were confirmed with LCT and colposcopy biopsy were considered. The patients were treated with LEEP treatment or cold knife conization. After treatment, all cases were strictly followed up with LCT and HPV test, and the patients with positive results were examined by histology. Results : 1) After treatment, there was no residual disease in all cases, the cure rate was 100%. 2) From 3 months after treatment, highrisk HPV negative rate was 100% in CIN1 cases. While at 3rd and 6th month after treatment, highrisk HPV negative rate in CIN2~3 cases were 7317% and 9085%, which were significantly lower than those in CIN1 cases,the difference was statistically significant. 3) From the 12th monthafter treatment, there are still two cases of sustained highrisk HPV positive but normal with LCT and colposcopy biopsy. All cases are still strictly followedup. Conclusion : After treatment, the negative rate and time of highrisk HPV concerned with the grade of the CIN; the patients with persistent positive highrisk HPV, but without abnormalities detected by LCT and colposcopy biopsy could continue to strictly follow up; LCT joint highrisk HPV detection is an effective clinical means for followup after CIN treatment.
Objective To summarize and review the heterogeneity of bone marrow derived stem cells (BMDSCs) and its formation mechanism and significance, and to analyze the possible roles and mechanisms in intestinal epithel ial reconstruction. Methods The related l iterature about BMDSCs heterogeneity and its role in intestinal epithel ial repair was reviewed and analyzed. Results The heterogeneity of BMDSCs provided better explanations for its multi-potency. The probable mechanisms of BMDSCs to repair intestinal epithel ium included direct implantation into intestinal epithel ium, fusion between BMDSCs and intestinal stem cells, and promotion of injury microcirculation reconstruction. Conclusion BMDSCs have a bright future in gastrointestinal injury caused by inflammatory bowl disease and regeneration.
【摘要】目的评价双氯芬酸钠滴眼液在准分子激光上皮下角膜磨镶术(LASEK)中的临床应用价值。方法2008年7月2009年3月行LASEK术患者80例160眼,屈光度范围-4.86D±2.15D(-1.25D～-6.50D),随机分为双氯芬酸钠滴眼液组(试验组)和对照组,对两组术后角膜刺激症状、上皮愈合情况进行对比观察和统计学分析。结果试验组术后第1、3天角膜刺激症状明显轻于对照组(P＜0.05),上皮愈合速度较对照组更快(P＜0.05)。结论在LASEK术中合理使用双氯芬酸钠滴眼液,可明显减轻术后患者角膜刺激症状,促进角膜上皮的更快愈合。
PURPOSE:To establish methods for cryopreservation of human retinal pigment epithelial cells (RPEs)and cell culture from thawing of frozen cells. METHODS:Primary cultured RPEs or its first or second passages,added with 10 dimetbylsulfoxide,were kept in --20℃ for 1 to 2 hours,and then further froze to -40~C over night before being placed in liquid nitrogen. The frozen cells were thawed in 60℃ within 2 minutes. Trypan blue staining and immunocytochemical staining with anti-human keratin were performed for cell viability and differentiation. The growth curve was also determined by calculating the total number of cells/well/day. RESULTS:The viable rate from frozen RPEs was 90%. No differences were observed for growth activity between cultures from frozen cells and controls. The cells were positive with anti-human keratin staining. The logarithmic growth phase was during I to 4 days and the doubling time yeas 1.55 days. CONCLUSION: Cryopreservation of RPEs in liquid nitrogen can maintain biological activities of cells with normal growth and features after thaw- ing. This will provide cell lines for in vitro experiments and possibly for cell banks for RPE transplantation for some fundus diseases. （Chin J Ocul Fundus Dis,1997,13:157-159）
Objective To observe the clinical features of congenital hypertrophy of retinal pigment epithelium （CHRPE）. Methods The clinical data of 13 CHRPE patients including visual acuity, slit-lamp microscope examination, indirect ophthalmoscope examination and fundus fluorescein angiography (FFA) were retrospectively analyzed. The patients, 9 males and 4 females, with the mean age of 27.8 years. Results All patients were unilateral, without systemic diseases and no subjective symptoms in majority. Only 30.77% of initial diagnosis was correct, other diagnosis include choroidal nevi, old chorioretinopathy or no diagnosis. The round or oval black lesion was found in ocular fundus of all patients, 7.69% was located on the optic disk, 46.15% was located on the inferior temporal retina, 30.77% was located on the superior temporal retina, 15.39％ was located on the inferior nasal retina. 92.31％ was pigmented CHRPE and 7.69％ was non-pigmented CHRPE. FFA showed blocked fluorescence and transmitted fluorescence in the lesion, few eyes were found dilated capillary vessel and fluorescent leakage on the late stage of FFA, most eyes had normal retinal vessels. Conclusion The isolated CHRPE is round or oval black lesion in ocular fundus which lack of subjective symptoms, mostly located on the peripheral retina; the FFA characteristics showed blocked fluorescence and transmitted fluorescence, and CHRPE often misdiagnosed as other disease, it should be combine the ocular fundus manifestation with the FFA to diagnose properly.