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.
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.
Objective To study the construction feasibility of a biodegradable artificial esophagus by the squamous epithelial cells and the myoblast cells seeded on the small intestinal submucosa(SIS) and to investigate the growth patternand angiogenesis of the co-cultured human embryonic squamous epithelial cells and the skeletal myoblasts in vivo. Methods The squamous epithelial cells and the myoblast cells were obtained from the 20-week aborted fetus. Both of their cellswere marked by 5-BrdU in vitro.The isolated cells were then seeded on the SIS and co-cultured in vitro for 24 hours, and then the compound of the cells and the SIS was transplanted into the subcutaneous tissue of the athymismus mice. The observation on the morphology and the cytokeratin AE3 and α-actin specified immunohistochemistry of the squamous epithelial cells and the myoblastcells was performed at each of the following time points: 3 days, 1 week, 2 weeks, and 3 weeks after transplantation. Results The morphological observation indicated that the cultured cells could penetrate into the small intestinal submucosa and form several-layered cell structures, and that the compound of the cells and the SIS could have angiogenesis within 2-3 weeks. The 5-BrdU specified immunohistochemical observation suggested that the cells growing in the small intestinal submucosa scaffold might be the cells transplanted.The cytokeratin AE3 specified and α-actin specified immunohistochemical studies demonstrated that the transplanted cells could differentiate in vivo. Conclusion It is possible to fabricate the framework of a biodegradable artificial esophagus with the epithelial cells and the myoblast cells seeded on the small intestinal submucosa.
目的:探讨胚胎发育不良性神经上皮肿瘤(DNT)的临床、影像及病理学特征、诊断及鉴别诊断.方法:回顾性分析8例胚胎发育不良性神经上皮肿瘤患者的临床和影像学资料,进行光镜和免疫组织化学染色观察,并获得6例的随访资料.结果:胚胎发育不良性神经上皮肿瘤男性7例,女性1例,年龄为5～19岁,平均年龄13岁,5例以癫痫小发作为主要临床表现,病变均位于幕上,以皮层为主,影像学检查均无明显的占位效应及瘤周水肿。肿瘤细胞主要由少突胶质样细胞(OLC)、神经元和星形细胞组成,4例伴有皮质发育不良。免疫组织化学结果为神经元及部分少突胶质样细胞呈嗜铬素A、突触素及S-100阳性表达;少突胶质样细胞呈胶质纤维酸性蛋白(GFAP)阴性表达,而星形细胞呈GFAP阳性表达;Ki-67抗原标记指数lt;1%。结论: 胚胎发育不良性神经上皮肿瘤为WHOⅠ级良性肿瘤,可结合临床、影像及病理学表现明确诊断,预后良好,无需放疗和化疗。
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.
Objective To detect the effects of cytokines on the expression of early growth response gene-1 (Egr-1) in cultured human retinal pigment epithelial (RPE) cells. Methods Immunofluorescence staining, Western blotting and reverse transcription polymerase chain reaction (RT-PCR) were used to detect and quantitatively analyze the expression of Egr-1 protein and mRNA in cultured human RPE cells which were exposed to stimulants, including 20 mu;g/ml lipopolysaccharide (LPS), 40 ng/ml tumor necrosis factor (TNF)-alpha;, 10 U/ml interferon (IFN)gamma;, 30% supernatant of monocyte/macrophage strain (THP1 cells) and the vitreous humor from healthy human eyeballs, for 0, 10, 20, 30, 40 and 60 minutes, respectively. Results The RPE cells stimulated for 0 minute revealed faint green fluorescence of Egr-1 in the cytoplasm. With exposure to the stimulants, the expressionof Egr-1 increased obviously and b green fluorescence was found in cytoplasm in some nuclei of RPE cells. Compared with the untreated RPE cells, after stimulated by 20 mu;g/ml LPS, 40 ng/ml TNFalpha;, 10 U/ml IFNgamma;, 30% supernatant of THP-1 cells and the vitreous humor, the approximate ultimate amplitudes of Egr-1 mRNA enhanced 1.9, 1.3, 14, 1.2, and 1.4 times, respectively; the greatest amplitudes of Egr-1 protein increased 3.4, 1.2, 1.7, 32, and 1.3 times, respectively. Conclusion LPS, TNF-alpha;, IFN-gamma;, supernatant of THP-1 cells and the vitreous humor can upregulate the expression of Egr-1 mRNA and protein in cultured human RPE cells, and induce its nuclear transposition, which suggests the activation of Egr-1.
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.
Objective To observe the mutation frequency and the characteristics of rentinitis pigmentosa (RP)1 gene in the Chinese patients with autosomal dominant (AD) RP or sporadic RP (SRP), and to evaluate their potential effects on the pathogenesis of RP. Methods Fifty-five members from 7 Chinese families with ADRP, 30 patients with SRP, and 75 healthy adults were recruited. Polymerase chain reaction (PCR) and direct DNA sequencing were used to detect the sequence mutation in the entire coding region and splice sites of RP1 gene. Univariate analysis and multivariate analysis were used to detect the effect of RP1 gene mutation sites on RP. Results Four coding sequence variants were detected in the codes of 852，872，921 and 939 at the exon 4 of RP1 gene. The R872H alteration, which was found in both ADRP families and patients with SRP, showed positive correlation with RP confirmed by the multivariate logistic regression analysis. The P903L alteration was only found in ADRP families but not in the patients with SRP or the healthy adults. Conclusions The R872H alteration in the RP1 gene is likely to increase the risk of RP, and may be a susceptible gene of RP. Whether the P903L alteration is a diseasecausing factor needs to be further studied.