Objective To observe the expression of miR-204 and 211 human embryonic stem cells (hESCs) differentiated into retinal pigment epithelial (RPE) cells. Methods RPE cells were derived from hESCs by natural differentiation method, and were identified. miRNA expression profiles and real-time polymerase chain reaction (RT-PCR) of miR-204 and 211 were generated from the following groups: hESCs, hESCs-derived cells containing pigmented foci, hESCs-derived RPE cells and human fetal RPE (hfRPE) cells. Results miRNA-204 was continuously upregulated throughout the entire differentiation process of hESCs to RPE cells. It increased 5.026 times in hESCs-derived cells containing pigmented foci compared to hfRPE cells; it was increased 3.337 times in hESCs-derived RPE cells compared to hESCs-derived cells containing pigmented foci; it increased 13.574 times in hfRPE cells compared to hESCs-derived RPE cells. miR-211 does not change during differentiation from hESC to RPE, but it increased 44.333 times in hESCderived RPE cells compared to hfRPE cells. miR-211 was the biggest difference in the miRNA expression pattern. In four cell types of hESCs, hESCs-derived cells containing pigmented foci, hESCs-derived RPE cells and hfRPE cells, RT-PCR showed the levels of miR-204 were 91.81plusmn;4.43, 2263.09plusmn;206.39, 5996.80plusmn;235.42, and 171676.45plusmn;999.82 respectively. miR-204 was significantly increased during the whole course (t=18.22, 20.66, 279.38;P＜0.001). The levels of miR-211 were 2.23plusmn;0.31, 129.33plusmn;3.75, 125.7592plusmn;4.78, and 16682.00plusmn;352.97 respectively. miR-211 was significantly increased from hESCs to cells containing pigmented foci and from hESCs-derived RPE cells to hfRPE (t=58.58, 81.24; P＜0.001). Conclusion There is a continuous change of miR-204 and 211 in differentiation of RPE cells from hESCs.
Retinal degeneration mainly include age-related macular degeneration, retinitispigmentosa and Stargardt’s disease. Although its expression is slightly different, its pathogenesis is photoreceptor cells and/or retinal pigment epithelial (RPE) cel1 damage or degeneration. Because of the 1ack of self-repairing and renewal of retinal photoreceptor cells and RPE cells, cell replacement therapy is one of the most effective methods for treating such diseases.The stem cells currently used for the treatment of retinal degeneration include embryonicstem cells (ESC) and various adult stem cells, such as retinal stem cells (RSC), induced pluripotent stem cells (iPSC). and mesenchyma1 stem cells (MSC). Understanding the currentbasic and clinical application progress of ESC, iPSC, RSC, MSC can provide a new idea for the treatment of retinal degeneration.
Objective To establish a safe, effective, and economic feeder-free culture system which is suitable for the culture of human parthenogenetic embryonic stem cells (hPESCs) in vitro. Methods hPESCs were cultured with mTeSRTMl medium (control group) and human foreskin fibroblasts-conditional medium (hFFs-CM) (experimental group). The growth status of hPESCs in both feeder-free culture systems were observed with inverted microscope. Alkaline phosphatase (ALP) analysis and karyotype analysis were used to study the biological characteristics of hPESCs. The expression of hPESCs pluripotent marker Oct-4 was analyzed by RT-PCR. Differentiation experiment in vivo and in vitro was applied to observe the differentiation potential of hPESCs into three germ layers. Results hPESCs had regular morphology with difficulty in differentiation in both culture systems. No obvious difference was observed in morphology and expansion speed of hPESCs between 2 groups. After subcultured for 15 passages in vitro, hPESCs in 2 groups could maintain normal female diploid karyotype 46, XX and pluripotency. The expression of Oct-4 mRNA was positive in 2 groups. hPESCs in 2 groups could form embryonic body in differentiation experiment in vitro and could develop into teratomas containing three germ layers in nude mice. Conclusion Feeder-free culture system of hFFs-CM can sustain the growth of hPESCs and keep hPESCs undifferentiated state for long. A feeder-free culture system of hPESCs is successfully established, which can support the growth of hPESCs, reduce the contamination from animals, decrease the cost of culture, and satisfy the clinical large-scale application.
Objective To observe the effect of transplantation of embryonic stem cell(ES) on neurological functional recovery of injured spinal cord in adult mouse. Methods The ES cells were cultured and induced in vitro. Fifty C57/BL6J mice were made animal model of semicut mice of T9，10. The ES cellderived neural precursors cells were transplanted into the vertebral canalaround injured spinal cord semi-cut mice. Twenty-eight C57/BL6J mice were randomly divided into three groups: sham operation group(group A,n=9), operation／cell group (group B，n=10), and operation／DMEM group(group C,n=9). RT-PCR analysis, X-gal staining and immunofluorescence were used to observe the cells survival and differentiation in the spinal crod. BBB test was performed to study functional improvement. Results ES cells induced and cultured in vitro displayed clonal growth with circle or ovoid shape and had one or more nucleoli. RT-PCR result showed that the induced ES cells expressed mRNA of Nestin and microtubuleassociated protein, but did not express glial fibrillory acidic protein(GFAP). There was statistically significant difference in BBB scoring between group A and groups B, C after operation (P＜0.01). There was statistically significant difference in BBB scoring at 1, 2 and 4 weeks of operation(P＜0.01), but no statistically significant difference at 6 and 8 weeks of operation between groups B and C(P＞0.05). The X-gal staining results werepositive in group B and negative in groups A and C. The immunoflurescence resultshowed neurofilament green fluor and no expression of GFAP in injured spinal cord region. Conclusion After transplantation, ES cellderived cells can survive, transfer into the injury position, and differentiate into neurons, but spinal cord function has no obvious improvement.
This is the first successful case expriences,a method of the procurement of the fetal cadavertic multiple argans for transplantation of the pancreas and thyroid-pararthyroid glands was produced. The liver,pancreas,duodenum,spleen,and both kidneys were harvested en bloc by a group of surgeons,and the right hem-ithyroid-parathyroid glands with pedicle of thd blood vessels wre removed by another group.The pancreas together with the spleen were transplanted to a patient having diabetes mellitus. The thyroid-parathyroid glands were given to another case with bypothyroidism and hypoparathyroidism.Both cases had good results.This method had dicreased the warm ischemia of the transplants,and could provide liver,pancreas,spleen,kidneys and thyroid-parathyroid glands to solve the problem of shortage of fetal organs.
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 determine whether transplanting olfactory ensheathing cells (OECs) is effective in controlling or reversing the deterioration caused by amyotrophic lateral sclerosis (ALS). Methods Between February 2003 and April 2006, 327 patients (241 males and 86 females) with probable or definite ALS (diagnosed according to the El Escorial criteria) received the OECs transplantation. Their ages ranged from 20 to 84 years (51.6±11.1 years). The duration of symptoms before surgical treatment was 4-8 months to 13 years (2.9±2.0 years). OECs were cultured and injected into pathological regions of the spinal cord and/or bilateral corona radiata of the brain; the patients were divided into three groups, group A (cord only,n=29), group B (cord and brain,n=6), and group C (brain only,n=292) based on the transplant sites. Results The patient’s neurological function was assessed both before and at 4 weeks after transplantation by using the Amyotrophic Lateral Sclerosis Functional Rating Scale (ALSFRS) of the ALS CNTF Treatment Study (ACTS). The scores were increased from 17.2±8.6 preoperation to 20.1±9.7 postoperation in group A (P＜0.05),from 24.2±6.8to 25.7±6.6 (P＞0.05) in group B, and from 20.3±8.6 to 22.0±9.4 (P＜0.001) in group C.There were no significant difference in increased ALSFRS scores among the threegroups (P＞0.05). The total improvement rate of neurological function was 77.1% (252/327). The result of electromyographic examination showed that spontaneouspotential diminished and/or disappeared, the amplitude of the motor unit actionpotential decreased remarkably and the numbers of motor unit action potential greatly increased in 261 cases (79.8%). Sixteen patients (4.9%) experienced thevarious complications including headache, shortterm fever, seizure attack, central nerve system infection, pneumonia, respiratory failure, urinary tract infection, heart failure, and possible pulmonary embolism; of them, there were 4 deaths(1.2%). Conclusion These preliminary results suggest that the OECs transplantation is effective in controlling or reversing the physiological deterioration caused by ALS.
Purpose To investigate the development of embryonic stem cells (ESC)in the subretinal space． Methods ESC were cultivated in suspension for 4 days till they developed into cell aggregates,i.e.embryonic body(EB).ESC as well as EB combined with or without RA were respectively transplanted into vitreous cavity and subretina1 space in SD rats,and the subretinal transplanted eyes,transient ischemia-reperfusion injuries were made by ligating the ophthalmic artery for 40 seconds before the transplantation .The experimental eyes were enucleated for histological and immunohistochemical assays after 14～28 d. Results The EB was found to develope into photoreceptors induced by RA in the subretinal space under an ischemia-reperfusion condition，and EB transplantation without RA induction induced multiple differentiations in the subretinal space.The single injection of RA without EB induced hyperplasia of the neural retinal cells.ESC transplanted into vitreous cavity rapidly proliferated and developed into atypical hyperplastic mass. Conclusion EB derived from ESC can differentiate into photoreceptors induced by RA in the host subretinal space under an ischemia-reperfusion condition. (Chin J Ocul Fundus Dis，2000，16：213-284)
ObjectiveTo systematically review the clinical effects of short-term and conventional fertilization for vitro fertilization-embryo transfer (IVF-ET). MethodsRandomized controlled trials (RCTs) about the clinical effects of short-term fertilization versus conventional fertilization for IVF-ET were searched in PubMed, The Cochrane Library (Issue 8, 2014), CBM, CNKI, WanFang Data and VIP from inception to August 2014. Two reviewers independently screened literature according to the inclusion and exclusion criteria, extracted data, and assessed methodological quality of included studies. Then meta-analysis was performed using RevMan 5.2 software. ResultsA total of six RCTs involving 1 373 patients were finally included. The results of meta-analysis indicated that:short-term fertilization was superior to conventional fertilization in increasing high quality embryo rates (OR=1.42, 95%CI 1.18 to 1.70, P=0.000 2) as well as clinical pregnancy rates (OR=1.67, 95%CI 1.33 to 2.09, P < 0.000 01). However, the two groups were alike in fertilization rates, polyspermy rates, and miscarriage rates. ConclusionCurrent evidence indicates that short-term fertilization is superior to conventional fertilization in increasing high quality embryo rates as well as clinical pregnancy rates. Due to limited quality and quantity of the included studies, the above conclusion should be verified by conducting more large-scale, high quality RCTs with long-term follow-up.
Objective To analyze the efficacy and safety of different acupuncture methods on outcome of in vitro fertilization-embryo transfer (IVF-ET). Methods The PubMed, EMbase, Cochrane Library, CNKI, VIP, WanFang Data and CBM databases were searched to collect randomized controlled trials (RCTs) related to the objectives of the study from the inception to April 16, 2023. After two investigators independently screened the literature, extracted the data and evaluated the risk of bias of the included studies, a network meta-analysis was performed using Stata 16.0 software. Results There were 62 trials total with 9844 patients, involving 7 interventions. Network meta-findings analysis revealed the following: ① Clinical pregnancy rate (CPR): needle warming > auricular acupressure > transcutaneous electrical acupoint stimulation (TEAS) > electroacupuncture > acupuncture > sham acupuncture > no adjunctive treatment; ② Live birth rate (LBR): electroacupuncture > auricular acupressure > TEAS > acupuncture > sham acupuncture > no adjunctive treatment. Conclusion Needle warming assisted IVF-ET is superior to other acupuncture therapies in improving CPR, especially during the promotion period of excretion, and the selection of Zusanli, Guanyuan and uterine acupoints for 3-month cycles may have the best effect. And for the LBR, the effect of electroacupuncture is better than that of other therapies. Besides, auricular acupressure may have good therapeutic potential. Due to the limited quality and quantity of the included studies, more high quality studies are required to verify the above conclusions.