非穿透性小梁切除加羊膜移植治疗开角型青光眼

目的 :探讨非穿透性小梁切除术术中加羊膜移植治疗开角型青光眼的机理及疗效。方法 :为 10例、12眼开角型青光眼行非穿透性小梁切除术联合术中巩膜瓣下植入羊膜 ,术后观察视力、眼压、滤过泡、眼内反应及并发症 ,随访 3～ 5个月。结果 :术后炎症反应轻 ,眼压控制良好 ,术后眼压为 14～ 19mmHg ,术后并发症少。结论 :非穿透性小梁切除术能安全、有效地降低眼压 ,是治疗开角性青光眼有效的手术方法。     

Amnion graft as a possible source of stem cells for endometrial regeneration after lysis of severe intrauterine adhesions

BackgroundThe current traditional management of severe intrauterine adhesions (IUAs) is hysteroscopic adhesiolysis with application of either an intrauterine contraceptive device or a Foley catheter balloon with reported recurrence in up to 50% of cases. Recently, significant improvement with the use of amnion graft following hysteroscopic lysis of severe IUA was reported.CaseIn the present case, endometrial stromal cells (ESCs) were found within and on the surface of amniotic membrane two months after its grafting intrauterine which was confirmed with CD10 immunohistochemistry.ConclusionAmniotic membrane graft after hysteroscopic lysis of severe IUA, might act as a source of stem cells for endometrial regeneration; a role that will be of great value especially in severe IUAs. Further studies to confirm this finding are warranted.     

羊膜移植在角膜溃疡的临床应用

目的 探讨羊膜移植治疗角膜病的临床效果.方法 采用保存羊膜联合清除角膜溃疡病灶及层间注药等方式治疗角膜溃疡32例,术后随访3～24个月.结果 32例角膜上皮均修复,无新生血管生长.11例视力提高,占34.38％;16例视力无变化,占50％;5例视力下降,占15.62％.结论 羊膜移植是治疗角膜疾病的有效方法.     

葛根芩连汤联合人羊膜间充质干细胞移植治疗糖尿病早期视网膜病变

BACKGROUND:Human amniotic mesenchymal stem cells have the potential to differentiate into insulin-secreting cells and have immunomodulatory effects. Additionally, it has been clinically proved that Gegen Qin Lian Tang can lower blood sugar. OBJECTIVE:To investigate the effects of Gegen Qin Lian Tang combined with human amniotic mesenchymal stem cell transplantation on rat early diabetic retinopathy, by detecting pathological changes, levels of blood indicators and expressions of vascular endothelial growth factor in retinal tissues. METHODS:By intraperitoneal injection of streptozotocin, 42 Wistar rat models with diabetes mellitus were prepared and then randomly divided into model group, human amniotic mesenchymal stem cell transplantation group (cell transplantation group) and combination of human amniotic mesenchymal stem cells and Gegen Qin Lian Tang group (combination group). At 4 weeks after transplantation, levels of blood sugar and serum insulin in diabetic rats were detected; pathological changes of the retina in diabetic rats were observed by hematoxylin-eosin staining. Besides, expressions of CD45 and vascular endothelial growth factor mRNA were detected using immunohistochemistry staining and RT-PCR technology, respectively. RESULTS AND CONCLUSION:Compared with the model group, the levels of blood glucose and serum insulin in both cell transplantation and combination groups were significantly decreased and increased, respectively, and these changes were even more significant in the combination group (P < 0.05). In the model group, the retinal edema could be found, accompanied by structural disorder and irregular cell arrangement, while these retinal lesions were relatively milder in the cell transplantation group and significantly improved in the combination group. In addition, the CD45 expression and in the retina was highest in the model group subsequently followed by the cell transplantation group (P < 0.05) and lowest in the combination group (P < 0.05). Furthermore, compared with the model group, the expression of vascular endothelial growth factor mRNA was also significantly lower in the other two groups, especially in the combination group (P < 0.05). These data show that Gegen Qin Lian Tang combined with human amniotic mesenchymal stem cell transplantation improves the severity of diabetic retinopathy by inhibiting expressions of CD45 and vascular endothelial growth factor in the retinal tissues.     

Chorioamniontis in preterm delivery is associated with degradation of decorin and biglycan and depletion of hyaluronan in fetal membranes

Objective

The proteoglycan decorin stabilizes collagen whereas biglycan and hyaluronan disrupt well-organized collagen. The aim was to determine the concentrations of these constituents in fetal membranes in relation to gestational age, preterm labour, PPROM and chorioamnionitis.

Study design

Preterm fetal membranes (24–34 weeks gestation) were obtained from elective caesarean deliveries (N = 4), from PPROM (N = 14), and from preterm labour (N = 14). Term fetal membranes from elective caesarean deliveries (N = 9) and spontaneous vaginal deliveries (N = 11) were used for comparison. Chorioamnionitis was assessed histologically. The proteoglycans were analysed using alcian blue precipitation, SDS-PAGE and immunostaining. Hyaluronan was estimated by a radioimmunoassay.

Results

Preterm amniotic membranes with chorioamnionitis displayed a 8-fold decrease in hyaluronan concentration as well as a pronounced (88%) degradation of decorin and biglycan (p < 0.05).The amnion from preterm elective caesarean sections had higher decorin (3.2 vs. 1.7 μg/mg, p < 0.05) and lower biglycan (0.4 vs. 1.0 μg/mg, p < 0.05) concentrations as compared to similar term amnion (p < 0.05), whereas the hyaluronan concentrations were not associated with gestational age. Also the chorio-decidua from preterm caesarean sections had higher decorin concentrations (1.8 vs. 1.0 μg/mg, p < 0.05) whereas the biglycan concentration was unchanged. Labour (term as well as preterm) was characterized by increased hyaluronan and biglycan concentrations in the amnion (not statistically significant).

Conclusion

The biglycan/decorin balance increases during third trimester of pregnancy and during active labour. This relation might contribute to mechanical weakening of the membranes. Chorioamnionitis induces dramatic degradation of both proteoglycans and hyaluronan, which can explain the decreased biomechanical strength.     

不同消化分离方法分离人羊膜间充质干细胞效果比较

目的： 比较不同消化分离方法提取人羊膜间充质干细胞（HAMSCs）的分离效果,以供选择一种更有效的分离方法。方法: 将新鲜羊膜组织分成4片6 cm×6 cm大小的组织块,采用4种不同方法进行消化分离：第1组：先刮除+胶原酶Ⅱ组;第2组：先刮除+胶原酶Ⅳ组;第3组：先剪碎+胶原酶Ⅱ组;第4组：先剪碎+胶原酶Ⅳ组。对这4组原代培养HAMSCs进行存活数、形态学观察比较,第3代 HAMSCs抗原检测及成脂成骨诱导分化实验。结果: 第1组和第2组的HAMSCs存活数高于第3组和第4组(P<0.05)。第1组和第2组的HAMSCs成份较第3组和第4组纯净,后2组夹杂着一些团状的人羊膜上皮细胞（HAECs）。抗原结果显示CD29、CD44、CD73、CD90、CD105表达阳性,CD31、CD34、CD45、HLA-DR表达阴性,Vimentin、SSEA-3、SSEA-4、OCT-4、telomerase结果阳性。成脂成骨诱导分化实验成功。结论: 通过先将人羊膜用胰蛋白酶+EDTA消化液消化后,再将HAECs刮除干净,然后剪碎再用胶原酶消化的改进方法对HAMSCs保护较好,较易提取,获取细胞量和纯度均较传统方法高。胶原酶Ⅱ和胶原酶Ⅳ2种消化酶对HAMSCs的提取影响不大。     

负载自体骨髓间充质干细胞羊膜片移植治疗翼状胬肉

目的观察负载自体骨髓间充质干细胞羊膜片移植治疗翼状胬肉的临床疗效。方法10例（10眼）原发性巨大翼状胬肉患者,在知情同意下,抽取自体骨髓间充质干细胞,将鉴定后的第3代骨髓间充质干细胞接种到羊膜上,切除翼状胬肉,将载有自体骨髓间充质干细胞的羊膜片细胞面向上覆盖在巩膜裸区缝合固定后,另缝合一层单纯羊膜保护移植细胞。结果10例患者移植术后1-3d角膜剥离区上皮修复,1周左右周围结膜上皮逐渐移行生长,10d至2周时羊膜片开始融合吸收。随访6月至3年,无复发。结论负载自体骨髓间充质干细胞的羊膜片移植治疗翼状胬肉,对结膜刺激小,眼表瘢痕轻,胬肉无复发。     

逆行切除加羊膜移植术治疗翼状胬肉65眼疗效观察

目的 观察胬肉逆行切除加羊膜移植术防止翼状胬肉复发的疗效.方法 将54例（65眼）翼状胬肉沿胬肉体两侧分别剪开球结膜,钝性分离球结膜与其下的增生组织,于胬肉体部剪断已分离的球结膜,并向泪阜侧掀开,将增生的胬肉组织充分暴露,再将胬肉与巩膜进行分离;于半月皱襞处剪断胬肉根部,然后逆行将胬肉头部自角膜上分离下来,将羊膜移植于巩膜裸露区.结果 65眼随访6～18个月,12眼复发,复发率为18.5%.结论 翼状胬肉逆行切除加羊膜移植术可有效降低翼状胬肉的术后复发率.