ObjectiveTo evaluate the safety and efficacy of amniotic membrane patching in the treatment of recurrent macular hole associated with retinal detachment of high myopia (MHRD). MethodsA prospective study. From March 2018 to January 2020, 11 patients (11 eyes) of recurrent macular hole associated with MHRD at the First Affiliated Hospital of Zhengzhou University were enrolled. Among them, there were 3 males (3 eyes), and 8 females (8 eyes). The average age was 63.64±5.82. The axis length (AL) was 29.10±0.59 mm, and the logarithm of the minimum angle of resolution best corrected visual acuity (logMAR BCVA) was 2.23±0.57. Patients previously received pars plana vitrectomy (PPV) combined with internal limiting membrane stripping surgery, which was more than 1 time. All eyes underwent standard pars plana three-channel 23G PPV combined with amniotic membrane covering and silicone oil filling. The silicone oil was removed 6 months after surgery. Follow-up time was up to 3 months after silicone oil removal surgery. 1, 3, and 6 months after the operation, the same equipment and methods were used to conduct relevant examinations before the operation to observe the closure of the macular hole, retinal reattachment and changes in logMAR BCVA. The logMAR BCVA before and after surgery was compared by paired t test. ResultsAt 1, 3, and 6 months after the operation, the retinas of all eyes were anatomically repositioned, the macular holes were well closed, and the amniotic membrane was attached to the retina. At 3 months after the silicone oil removal operation, there was no recurrence of macular hole in all eyes; logMAR BCVA was 1.35±0.32. No serious complications occurred during and after surgery in all eyes. ConclusionAmniotic membrane patching is a safe and effective method for recurrent macular hole associated with MHRD.
目的:观察新鲜羊膜移植联合丝裂霉素C治疗复发性翼状胬肉的临床疗效。方法:对32例(38眼)复发性翼状胬肉行翼状胬肉切除联合新鲜羊膜移植加丝裂霉素C治疗,观察术后角膜上皮愈合、胬肉复发情况。结果:术后随访3～24个月,有2眼复发,复发率为5.26%。结论:新鲜羊膜移植联合丝裂霉素Ｃ治疗复发性翼状胬肉降低了复发率,无严重手术并发症,是一种安全有效的手术方法。
Objective To investigate the effects of human acellularamnion membrane on SD rat tendon adhesion and to obtain the experimental data for clinical application in preventing postoperative tendon adhesion. Methods The tendons of 28 adult SD rats hindlimb were cut and sutured. The tendons of left hindlimb were encapsulated by human accellular amnion membraneas the experimental group and the ones of the other side were not encapsulatedas control group. The rats were killed 1, 2, 4, 6, 8 and 12 weeks after operation. The results were evaluated grossly and histologically. Results There were no differences in healing of injury tendon and inflammatory response between the two groups. The anatomical and histological results showed the experimental group had less adhesion than the control group(Plt;0.05). Conclusion Human acellular amnion membrane can prevent adhesion of tendonwithout affecting tendon healing and is an optimal biological material to prevent tendon adhesion.
Objective To observe the effects of culture medium of amniotic cells on NO and NOS in retinal tissues of rabbits in vitro in order to provide a protective method for antioxidation in retina transplantation. Methods Thirty adult healthy rabbits (30 right eyes) were divided into 3 groups. Group I: fresh retinal tissue; group II: routine culture medium; group III: culture medium of amniotic cells. The retinal tissues in group II and III were cultured in the corresponding culture medium for 1 week. The content of NO and NOS in retinal tissues in the 3 groups were determined. Results Compared with group I, the content of NO and NOS of group II increased obviously (t=3.821, 3.854; P＜0.001). There was no statistical difference of content of NO and NOS between group I and III (t=1.657, 1.745; P＞0.05). Conclusion Culture medium of amniotic cells may remove free radicals and enhance the ability of antioxidation. (Chin J Ocul Fundus Dis，2004，20：366-368)
截止至2002年5月,现有早产治疗的临床证据如下: (1) 高危早产:在一些国家实施的RCT发现,在降低早产危险方面,加强产前保健与普通产前保健没有明显差异.包括5个RCT的1个系统评价发现,对有宫颈改变的妇女行宫颈环扎术有不同的结果,没有明确的结论.1个大样本的RCT发现,孕9～29周宫颈功能可能不全的妇女进行预防性宫颈环扎手术与不环扎相比,能明显降低早产(＜33孕周),但也会明显增加产褥感染的危险.另外4篇较小样本的RCT发现,孕10～30周、具各种早产高危因素的妇女,进行预防性宫颈环扎手术与不环扎相比,并不能降低早产(＜34孕周).1篇系统评价的2个RCT报告,对有宫颈改变的妇女进行环扎术有不同的结果,其中1个RCT发现其并不能明显降低早产(＜34孕周),而另外1个较小样本的RCT却发现宫颈环扎手术加卧床休息与单纯卧床休息比较,能明显降低34周前的早产.没有1个RCT证实行环扎术加卧床休息与单纯卧床休息相比,能降低围生儿死亡率. (2) 胎膜早破:1个系统评价发现,对胎膜早破的妇女,抗生素较安慰剂能明显延长孕周、降低新生儿发病率的危险,如新生儿感染、出生后氧疗、脑部超声异常等.阿莫西林加克拉维酸治疗与新生儿坏死性小肠结肠炎的发生率明显增加有关.一个基于1个RCT的系统评价发现,没有充足的证据证实羊膜腔灌注与不灌注比较能改善胎膜早破后的新生儿结局. (3) 先兆早产的治疗:①β-肾上腺素兴奋剂:1个系统评价发现,β-肾上腺素兴奋剂与安慰剂或不治疗相比,并不能明显降低围生儿死亡率、呼吸窘迫综合征及低体重儿(＜2 500 g)发生率,且与与安慰剂或不治疗相比,β-肾上腺素兴奋剂增加孕母副反应,如胸痛、心悸、呼吸困难、震颤、恶心、呕吐、头痛、高血糖、低钾血症.②钙离子通道拮抗剂: 没有关于钙离子通道拮抗剂与安慰剂比较的系统评价或RCT.1个系统评价发现,钙离子通道抑制剂与其它保胎药(主要是β-肾上腺受体兴奋剂)比较,能显著降低48 h内的早产分娩,减少因孕母副反应退出治疗和新生儿发病率.③硫酸镁:1个系统评价发现,硫酸镁与安慰剂比较,并不能明显降低孕36周前的早产率、围生儿死亡率、呼吸窘迫综合征的发生率.另一个系统评价发现,硫酸镁和其他宫缩抑制剂(β-肾上腺素兴奋剂、钙离子通道拮抗剂、前列腺素合成抑制剂、硝化甘油、酒精和葡萄糖注射剂)比较,并不能明显降低48 h内早产率(尽管结果没有差异).④垂体受体拮抗剂(阿托西班):1个系统评价纳入 2个RCT,对阿托西班和安慰剂治疗早产进行比较有不同的结果.较大样本的RCT发现,阿托西班较安慰剂能延长孕周,但阿托西班增加了孕28周以下的胎儿死亡率.另一个RCT发现,阿托西班增加了48 h内的早产.⑤前列腺素抑制剂(消炎痛):1个系统评价发现,消炎痛与安慰剂比较,能明显降低孕37周前的48 h和7天的早产率的证据有限.然而,同时发现消炎痛与安慰剂或不治疗相比,并不能明显降低围生儿死亡率、新生儿呼吸窘迫综合征、肺支气管发育不良、坏死性小肠结肠炎、新生儿败血症或低体重儿.但这个系统评价样本太小,尚不能发现有临床意义的差异. (4) 择期或非择期剖宫产对早产妇女治疗效果:1个系统评价结果发现,择期剖宫产较非择期剖宫产会增加孕母的发病率,却不能降低新生儿的发病率和死亡率.但尚不能证明此效果是否对新生儿有临床意义. (5) 改善早产妊娠结局的干预措施:①对早产者采用皮质类固醇:1个系统评价认为,对可能发生早产的妇女使用皮质激素较安慰剂或不处理能明显降低早产儿出生后呼吸窘迫综合征、新生儿死亡率和颅内出血的发生.②促甲状腺激素释放激素在早产中的运用:1个系统评价发现,在早产的高危妇女中,促甲状腺激素释放激素和类固醇激素联合应用与单用皮质类固醇激素比较,对新生儿结局的影响无明显差异,但会明显增加孕母和胎儿的不良反应.③抗生素:1个系统评价发现,抗生素与安慰剂比较,不能延长孕周、降低新生儿死亡率,但可降低孕母感染率.
Objective To study the differentiation of the human osteoblasts during the construction of the tissue engineered periosteum with the human acellular amniotic membrane(HAAM).Methods To construct the tissue engineered periosteum (n=60) with HAAM, the human fetal osteoblasts were used. The fetal osteoblasts were cultured for 2, 4, 6, 8, and10 days, and then their total RNA was extracted, which were reversely transcripted to cDNA. The realtime PCR analysis was used to reveal Cbfal and Osterix, and the cycle threshold (Ct) was also measured. The simplycultured osteoblasts were used as the control group (n=20).Results The expression of Cbfa1 was higher in the experimental group on the 2nd day when compared with that on the 4th, 6th, and 8th day(P＜0.05). The same result existed on the 10th day when compared with that on the 4th and 8th day. The expression of Osterix increased and was highest on the 8th day when compared with the other results(P＜0.05). Both of the 2 gene expressions were decreased in the control group when compared with those in the experimental group, but with no significant difference(P＞0.05). Conclusion Cbfa1 and Osterix can be normally expressed by the osteoblasts after their integration with HAAM. As a scaffold, HAAM can be used to keep the osteoblast phenotype and differentiation with an osteoconductive ability. Such a cell-scaffold complex may provide a basis for the osteogenesis.
Objective To prepare human acellular amniotic membrane(HAAM) and to measure its cytocompatibility and biocompatibility. Methods HAAM were preparedby chemical detergent-enzymatic extraction. Fresh human amnion was crosslinkedwith glutaradehyde, shaken in 0.5% SDS for 24 hours, and then treated with 0.25%trypsin for 4 hours. The production were freeze-drying and sterilized using ethylene oxide. Human fibroblasts were isolated from embryo and expanded in vitro. The fibroblasts were seeded in HAAM. HAAM and specimen were stained with HE and Mallory, and observed grossly, under light microscopy and scanning electron microscopy. The HAAM were implanted in the back of SD rats. Results There wereno residues of cells in the HAAM (HE, Mallory staining). One side of HAAM had reticular and porous structure, the other side had compact fibrous structure.Pore size was from 10 to 80 nm. The HAAM could be seeded with expanded fibroblasts in vitro,and fibroblasts had the potential of spread and proliferation. The SD rat in the implant test had no death, convulsions and other abnormal response. Conclusion The detergent-enzymatic extraction process can remove cellsand solvable components effectively and preserve the tissue matrix well and keep the reticular structure. The HAAM can be used as an ideal scaffold of biological membrane for tissue engineering.
Objective To investigate whether human amniotic mesenchymal stem cells (hAMSCs) have the characteristics of mesenchymal stem cells (MSCs) and the differentiation capacity into ligament fibroblastsin vitro. Methods The hAMSCs were separated through trypsin and collagenase digestion from placenta, the phenotypic characteristics of hAMSCs were detected by flow cytometry, the cytokeratin-19 (CK-19) and vimentin expression of hAMSCs were tested through immunofluorescence staining. The hAMSCs at the 3rd passage were cultured with L-DMEM/F12 medium containing transforming growth factor β1 (TGF-β1) and vascular endothelial growth factor (VEGF) as the experimental group and with single L-DMEM/F12 medium as the control group. The morphology of hAMSCs was observed by inverted phase contrast microscope; the cellular activities and ability of proliferation were examined by cell counting kit-8 (CCK-8) method; the ligament fibroblasts related protein expressions including collagen type I, collagen type III, Fibronectin, and Tenascin-C were detected by immunofluorescence staining; specific mRNA expressions of ligament fibroblasts and angiogenesis including collagen type I, collagen type III, Fibronectin, α-smooth muscle actin (α-SMA), and VEGF were measured by real-time fluorescence quantitative PCR. Results The hAMSCs presented monolayer and adherent growth under inverted phase contrast microscope; the flow cytometry results demonstrated that hAMSCs expressed the MSCs phenotypes; the immunofluorescence staining results indicated the hAMSCs had high expression of the vimentin and low expression of CK-19; the hAMSCs possessed the differentiation ability into the osteoblasts, chondroblasts, and lipoblasts. The CCK-8 results displayed that cells reached the peak of growth curve at 7 days in each group, and the proliferation ability in the experimental group was significantly higher than that in the control group at 7 days (P<0.05). The immunofluorescence staining results showed that the expressions of collagen type I, collagen type III, Fibronectin, and Tenascin-C in the experimental group were significantly higher than those in the control group at 5, 10, and15 days after culture (P<0.05). The real-time fluorescence quantitative PCR results revealed that the mRNA relative expressions had an increasing tendency at varying degrees with time in the experimental group (P<0.05). The relative mRNA expressions of collagen type I, collagen type III, Fibronectin, α-SMA, and VEGF in the experimental group were significantly higher than those in the control group at the other time points (P<0.05), but no significant difference was found in the relative mRNA expressions of collagen type I, collagen type III, and VEGF between 2 groups at 5 days (P>0.05). Conclusion The hAMSCs possesses the characteristics of MSCs and good proliferation ability which could be chosen as seed cell source in tissue engineering. The expressions of ligament fibroblasts and angiogenesis related genes could be up-regulated, after inductionin vitro, and the synthesis of ligament fibroblasts related proteins could be strengthened. In addition, the application of TGF-β1 and VEGF could be used as growth factors sources in constructing tissue engineered ligament.
ObjectiveTo observe the survival, migration, and effect of human amniotic epithelial cells (hAECs) on hepatic fibrosis in immune rats so as to provide the experimental theory for the clinical treatment with hAECs. MethodsSixty-four 10-week-old male Sprague Dawley rats (weighing, 220-280 g) were randomly divided into 4 groups, sixteen rats in each group. Rat hepatic fibrosis model was induced in groups A, B, and C; hepatic fibrosis rats were injected with 4×106 hAECs in group A, and with normal saline in group B, and no treatment was given in group C; group D served as control group. After 2 weeks of transplantation, the expression of human Alu gene repeat sequence was detected by DNA-PCR method and human leucocyte antigen G (HLA-G) by immunohistochemical staining in heart, liver, spleen, kidney, lung, and brain in group A, and then the percentage of positive expression was compared between organs except spleen. Semi-quantitative analysis was done for liver fibrosis with HE staining according to Chevallier semi-quantitative histological liver fibrosis scoring system, and immunohistochemical staining for TGF-β1 was used to record immunohistochemical score (ISH), the concentrations of aspartate transaminase (AST), alanine aminotransferase (ALT), and albumin (ALB) were determined to analyze hepatic fibrosis. ResultsAlu gene repeat sequence and HLA-G could be detected in liver, heart, brain, lung, and kidney in group A, the percentage of positive expression in the liver was significantly higher than that in the other organs (P<0.05). The histological semi-quantitative score of group A (10.47±3.20) was significantly lower than that of groups B and C[(13.84±3.46) and (13.85±3.16)](P<0.05), but no significant difference was found between groups B and C (P>0.05). The ISH scores in groups A, B, C, and D were 3.60±1.50, 5.38±2.60, 5.50±2.40, and 1.87±1.36, respectively; groups A, B, and C were significantly higher than group D, and group A was significantly lower than groups B and C (P<0.05), but there was no significant difference between groups B and C (P>0.05). The concentrations of ALT and AST in groups A, B, and C were significantly higher than those in group D, and group A was significantly lower than groups B and C (P<0.05), but there was no significant difference between groups B and C (P>0.05). The concentration of ALB in groups A, B, and C was significantly lower than that in group D, and group A was significantly higher than groups B and C (P<0.05), but there was no significant difference between groups B and C (P>0.05). ConclusionhAECs can survive in immune rats by intrasplenic transplantation and migrate to liver, heart, brain, lung, and kidney, and the liver shows the largest migration. The transplantation of hAECs in immune rat with cirrhosis can alleviate hepatic fibrosis and improve the serum indexes of liver function.
Objective To review the latest development of amniotic membrane andits application. Methods Related literatures on the development of amniotic membrane and its application were extensively reviewed and summarized. Results There were amniotic epithelial cells and many growth factors in the outer layer of amniotic membrane and there were many kinds of collagen in the basement. The special structure promoted the growth of many kinds of cells. It was widely used in ophthalmology. Conclusion As it is easily available, compatible, cheap in price, low in antigenicity, and able to promote the growth of many kinds of cells, with few ethical problems involved, amniotic membrane will be more and more widely applied.