Objective To study the effect of different types of supernatants fluid of retinal cells on the physiological function of neuron cells derived from embryonic stem cells. Methods Embryonic bodies were sub-induced by retinoic acid (group A), retinoic acid with the supernatant fluid of retinal glia cells and neurons of mouse (group B), retinoic acid with the supernatant fluid of fetal retinal glia cells (group C), respectively. The Sodium ion channels on the cytomembrane in the 3 groups were analyzed 5-21 days after the inducement. Results The sodium current in each group didn't change much 5-21 days after the inducement. The sodium channels presented burst-opening discharge in group A, brief-opening discharge in group B, and long-opening discharge in group C. The percentage of the cells without current in group A, B and C was 25%, 11.4%, and 23.8%, respectively, but the difference was not significant among the 3 groups(Pgt;0.05). The number of cells with sodium current increased at first and decreased later in group A, continuously increased in group B, and decreased at first and kept stable later in group C. The open time of sodium channels was the longest in group A, and the shortest in group B. The distribution of open time in the three groups could be managed with two-step exponential fit. Conclusion The supernatant fluid of retinal cells has apparent influence on the physiological function of the neuron cells derived from embryonic stem cells. (Chin J Ocul Fundus Dis, 2007, 23: 91-93)
Objective To observe the distribution of human photoreceptor cells at the posterior pole, detect the change of density of the cells affected by eccentricity, and analyze the relationship between the density distribution and the visual sensitivity. Methods Twenty human eye cups with the cornea removed were fixed in 4% polyformaldehyde for 1-4 weeks, and the retinal mounts were observed by differential interference contrast microscope to reveal the retinal cellular configuration and density. The inner segments of photoreceptor cells were first observed from the center to the temporal peripheral part of the retinal mounts. Results The highest density of visual cone cells was at the central fovea (134 000-267 000/mm2, mean 198 090/mm2; CV value:18.2%). The density and individual variation decreased rapidly in the peripheral area. The high density area of rod cells was at the 4 mm of the eccentricity, with the highest value of 72 610-182 350/mm2 and with the high density between 3 and 5 mm. Conclusions The inner segment of photoreceptor cells was monolayer, which may tell the cellular absolute value. The high density of retinal cone cells at the central fovea provide the basis of sensitive central visual acuity, which relates to the individual variation and development. The rod cells have the peak density at the eccentricity with 4 mm, and this area has the greatest sensitivity of dim vision.
Objective To systematic review of bladder cancer antigen (BTA) stat and urine cytology (UC) in the diagnosis of bladder cancer. Methods MEDLINE (Jan.1966 to June 2008), EMbase (Jan.1988 to June,2008), Cochrane Library (Issue 1,2008), CMCC (1979 to June, 2008) and CNKI (Jan.1979 to June, 2008) were searched for studies about BTA stat and cytology in the diagnosis of bladder cancer. The search strategy was made according to the Collaborative Review Group search strategy. Quality of included trials wa assessed by quality assessment of diagnostic accuracy studies.Data were extracted by two reviewers using the designed extraction form. The software MetaDiSc1.4 was used to review management and data analysis. Results In total, 71 relevant studies were searched, of which 13 were included and 58 were excluded, with 3 733 patients involved. Heterogeneity (except for threshold effect) was found within these studies. A meta-analysis was performed using random effect model. Pooled accuracy indicators of sensitivity, specificity, positive likelihood ratio (LR) , negative LR and diagnostic odds ratio (dOR) and 95%CI of BTA stat and UC were 0.68 (0.65,0.70), 0.74 (0.72, 0.76), 2.51 (2.04, 3.09), 0.46 (0.38, 0.55), 5.66 (3.87, 8.29) and 0.41 (0.39, 0.44), 0.97 (0.97, 0.98), 12.64 (7.58, 21.08), 0.62 (0.55, 0.71), 22.16 (12.38, 39.66), respectively. The sensitivity of both methods increased as the higher of tumor grade and stage, and the incipient tumor was higher than the recurrence. Area under curve (AUC) of SROC curve of BTA stat and UC were 0.753 5 and 0.711 9, and Q index were 0.696 3 and 0.662 4, respectively. Conclusions The performance of urine BTA stat is moderate in the diagnosis of bladder tumor. It can not replace the traditional urine cytology and diagnose the bladder cancer alone, but which can be an available noninvasive examination and an important adjunct of preoperative detecting and postoperative monitoring of bladder tumor.
Objective To study the progressive development of the retinas through an observation on the histological changes of the retinas from neonatal mice of different day-ages. Methods The retinas from the mice of 1 to 20 days of age were examined by light microscopy,and from 1 to 3 days,by autoradiography. Results The retinas of the mice below 3 days of age only had the RPE cells layer,the neuroblast layer and the ganglion cell layer.With the increase in dayage,the retinas developed gradually and would be mature in the 20th day. Conclusions The retinas of mice is a kind of immature tissue before the 20th days,so it can be considered as transplantation donors. (Chin J Ocul Fundus Dis, 1999, 15: 174-176)
The human wild-type Rb cDNA has been inserted into a retrovirus vector DOL and introduced into the human breast cancer cell MDAMB468,which has a large deletion of exons 3-27 of Rb genes,by electroporation transfection techniques.The exogenous Rb gene expresses the 110kd Rb protein.The morphology of the transfected cells is similar to that of the parent MDAMB468 cells.With the expression of Rb protein,the growth rate of MDAMB468 cells decreases by about 50%,and the colony formation ability in soft agaris repressed completely.After injection of 3times;106Rb+ cells and Rb-MDAMB468 cells into nude mice,the tumors formed from 106Rb+ cells are smaller than those from Rb-cells.The cell population of G1 and S phase of Rb+ MDAMB468 cells increases and the proliferation quotient decreases by about 50%.This result supports the former report the Rb protein. (Chin J Ocul Fundus Dis,1993,9:135-140)
目的 探讨纤维乳管镜(FDS)联合液基薄层细胞学检查(TCT)在诊断非血性乳头溢液中的临床价值。方法 回顾性分析笔者所在医院2008年7月至2011年7月期间因非血性单侧单孔乳头溢液的176例住院患者的临床资料。所有患者均行FDS检查及冲洗液TCT,且均有明确的病理学诊断结果,比较联合法与单一FDS或TCT检查的诊断准确率。结果 FDS的诊断准确率为80.7%(142/176),TCT为53.4%(94/176),联合法为86.9% (153/176)。结论 FDS联合TCT对非血性乳头溢液的诊断准确率高,可以提高FDS检查阴性的乳腺疾病的检出率。
Objective To observe the survival of human umbilical cord derived mesenchymal stem cells (hUC-MSCs) after injection into the vitreous of rabbits,and the animal safety under those procedures.Methods Twentyseven pigmented rabbits were randomly divided into 3 groups (intravitreal injection 1 week group,2 weeks group and 4 weeks group), each with 9 rabbits.For each animal the right eye was the experimental eye receiving hUCMSCs injection,while the left eye was the control eye receiving culture medium. The rabbit eyes were examined by slitlamp microscope, indirect ophthalmoscopy, fundus photography, fundus fluorescence angiography(FFA)and Tonopen tonometer before and after injection. hUCMSCs were labeled by CMDil in vitro, and their survival status was measured by confocal fluorescence microscopy, light microscope and transmission electron microscope at 4 weeks after injection. Results Four weeks after injection, a large number of the hUCMSCs were still alive in the vitreous cavity. The overall condition of those rabbits was good. The anterior segment and retina of experimental eyes were normal, without hyperfluorescence, hypofluorescence and leakage in the retina at 1,2 and 4 weeks after injection. There was no significant difference on IOP before and after injection at different time points (P>0.05), and no obvious changes at cornea, anterior chamber angle,lens,retinal structure by.light microscope and transmission electron microscope examination.Conclusion hUC-MSCs can survive in the rabbit vitreous for four weeks;intravitreal injection of hUCMSCs was safe and feasible.
目的:机械分离、培养小鼠耳蜗螺旋神经元,并进行免疫荧光细胞学鉴定,为后期进一步的实验研究提供实验材料。方法:采用初出生1~5天以内的昆明小鼠进行解剖、机械分离以获得螺旋神经节组织,进行原代培养后,应用神经微丝蛋白(Neurofilament protein,NFP-H)单克隆抗体进行免疫荧光细胞学鉴定。结果:机械分离后获得的螺旋神经节组织中的螺旋神经元,在体外培养条件下可以存活并进行正常分化。典型的螺旋神经元,其细胞形态呈椭圆形,胞体透明光滑、接近生理形态。荧光染色标记后,胞体和神经突起均显色好,Schwann细胞和成纤维细胞未着色。结论:应用机械分离的方法获得小鼠耳蜗螺旋神经节组织并进行培养,耳蜗螺旋神经元在体外可以稳定地存活生长。培养获得的细胞形态和生存状态接近生理状态,满足电生理、免疫细胞化学、药理学等研究。应用特异性的神经微丝蛋白对培养获得的螺旋神经元进行免疫荧光细胞学鉴定,特异性好,荧光显色好。
Objective To establish a culture method for human fetal retinal progenitor cells (RPC) in vitro. Methods Retinal neuroepithelium of 8-12-week human fetal were isolated and cultured in suspension and adherent methods. The passage cells were cultured and differentiated for 14 days with 5% fetal bovine serum without basic fibroblast growth factor (bFGF). The expressions of RPC and retinal final cells markers before and after the differentiation were detected by immunohistochemical analysis. Results The isolated cells cultured in suspension method congregated as the neurospheres and expressed the neuroectodermal marker nestin, but failed in passage and expansion; while the expression of nestin and serial passage were found in the cells cultured in adherent way. The differentiated passage cells expressed retinal final cells markers including glial fibrillary acid protein, beta;-tubulin and recoverin. Conclusions RPC derived from human fetal neural retina at the 8th12th week of gestation are capable of expansion and multipotentiality. (Chin J Ocul Fundus Dis, 2007, 23: 98-100)
Objective To observe the differentiation effect of rabbit amnion-derived stem cells (ADSC) induced into neural cells.Methods ADSC of New Zealand female rabbits were isolated and cultured. Its mRNA level of Fibronectin, Nestin and Vimentin were detected by real-time quantitative polymerase chain reaction. The selfreplication ability of ADSC was confirmed by monoclonal formation experiments. These ADSC were further induced into neural cells in vitro. Five days after induced differentiation, the expression of -tubulin and glial fibrillary acidic protein (GFAP) were detected by immunofluorescent staining. Results ADSC were separated from amnion tissue gradually after 24 hours. There were polygonal cells gathered around the amnion tissue at 72 hours, and were distributed compactly around the amnion at 120 hours. The morphology of cleavage daughter cells was basically the same as parent cells. ADSC has the ability of self-replication. The Nestin, Vimentin, Fibronectin mRNA expressions in ADSC were 15.79, 1.91, 7.65 times those in spleen cells. The differences were statistically significant(Z=-5.243, -3.972, -2.524; P<0.05). The beta;-tubulin expression was found in cytoplasm of most cells. The GFAP expression was found in cytoplasm in some cells. Conclusions ADSC has self-replication ability. It can be induced into neurons and neuroglial cells under the right conditions.