Background Retinal pigment epithelial (RPE) cell transplantation is a novel approach to the treatment of hereditary retinal diseases, however, human-derived RPE cell line occurs de-differentiation during in vitro cell culture.Studies showed that early abnormal activation of Akt/mammalian target of rapamycin (mTOR) signal pathway is the primary cause of RPE cell line de-differentiation, therefore, the inhibition of mTOR pathway will be helpful for the retard of de-differentiation of RPE cells.Objective This study aimed to investigate whether rapamycin can suppress the activation of mTOR pathway and promote differentiation of ARPE19 cells.Methods ARPE-19 cells were incubated in 12-well plate and divided into control group and rapamycin-treated group.DMSO or rapamycin with the final concentration of 400 nmol/L was added in the medium of the control group and the rapamycin-treated group, respectively.The cells of each group were collected 24 hours and 48 hours after cultured.The expression of zonula occludens-1 (ZO-1) in the cells was examined by immunofluorescence.The relative expression levels of RPE cell specific genes and proteins were assayed by using real-time quantitative PCR and Western blot, respectively.The detected results were compared between the two groups.Results ZO-1 was expressed in both group,but the fluorescence intensity was evidently enhanced in the rapamycin-treated group.The relative expression levels of RPE65, MERKT and LRAT mRNA in the cells increased by 25.97％ , 29.71％ and 13.00％ in the rapamycin treated group compared with the control group 24 hours after cultured (P=0.04,0.04,0.04) , and the expression levels of RPE65, LRAT, rLBP1, BEST1 , keratin18 and MERKT mRNA elevated by 174.00％ , 88.00％ , 56.18％ ,193.81％ ,10.83％ and 35.02％ in the rapamycin-treated group in comparison with the control group 48 hours after cultured (P =0.00,0.04,0.01,0.04,0.04,0.03).In addition, the expressions of p-mTOR, p-P70S6 and p-S6 protein were weaker in the rapamyein-treated group than those in the control group both 24 hours and 48 hours after cultured.Twenty four hours after cultured,the expression level of ZO-1 protein raised by 40％ in the rapamycin-treated group compared with the control group (P =0.01);while 48 hours after cultured,the expression levels of ZO-1 ,MERKT, catenin and LRAT proteins elevated by 36.00％ ,57.37％, 13.68％ and 41.07％ in the rapamycintreated group in comparison with the control group (P=0.01,0.00,0.04,0.04).Conclusions Rapamycin can suppress the activation of mTOR signaling pathway and up-regulate the expressions of RPE specific genes in ARPE19 cells.Inhibition of mTOR pathway might be an effective way for culturing RPE cells in vitro.

Objective To investigate the effects of Schisandrae Sphenantherae Fructus and processed with vinegar on lipid metabolism of rats with type 2 diabetes mellitus (T2DM). Methods The rat model of T2DM was induced by high fat diet plus STZ. The rats were randomly divided into blank group, model group, Schisandrae Sphenantherae Fructus high-dose group and low-dose group, and vinegar Schisandrae Sphenantherae Fructus high-dose group and low- dose group. The rats in each group were fed with the corresponding medicine for gavage for 30 d. FINS, FFA, TC, TG, HDL-C, LDL-C, and MDH, total protein content of liver tissue were detected. HE staining was used to observe the histomorphological changes of liver and pancreas in rats. Results Compared with the model group, Schisandrae Sphenantherae Fructus groups and vinegar Schisandrae Sphenantherae Fructus groups did not show obvious effects on improving FBG and FINS, but it could raise varying degrees of HDL-C and MDH, and reduce FFA, LDL-C, TC, and TG, among which vinegar Schisandrae Sphenantherae Fructus could significantly regulate metabolism in T2DM rats. Conclusion Vinegar Schisandrae Sphenantherae Fructus can enhance the lipid metabolism regulatory function of T2DM rats.

Objective To explore the value of CT spinal angiography with 256 MSCT and fast dynamic contrast-enhanced 3D MR angiography (CE-MRA) at 3.0 T in the diagnosis of spinal vascular malformations by comparing with results of DSA and operation.MethodsSeventeen patients suspected of spinal vascular diseases by initial MR and clinical manifestations all underwent CT spinal angiography.Of them,10 patients underwent MRA,15 patients underwent DSA within 3-5 days,and 8 patients finally underwent surgical treatment.ResultsCTA examination clearly showed the abnormal vascular lesions in 16 of 17 cases,including 7 cases with the diagnosis of spinal dural arteriovenous fistula,7 cases of perimedullary arteriovenous fistula,and 2 cases of spinal arteriovenous malformations. The results were consistent with the diagnosis of DSA or surgery.One case was poorly diagnosed.The feeding vessels were correctly determined in 12 cases,and the level of fistulas were correctly displayed in 12 cases.The level of fistulas and feeding vessels were accurately showed in 7 of 10 cases with MRA,while the other 3 cases exhibited normal with DSA.ConclusionsSpinal angiography with 256 MSCT and CE-MRA at 3.0 T can clearly show the extent of spinal vascular malformations,feeding arteries and fistula location.They are safe,noninvasive,convinient and can shorten the time of DSA diagnosis and treatment.They play an important role in diagnosis and treatment of spinal vascular malformations and postoperative follow-up.

Background Retinal pigment epithelium (RPE) cell transplantation is the primary means of human trial for the treatment of retinal degeneration.Induced pluripotent stem cells (iPSCs) will become an important source for cell transplantation.In addition,iPS-RPE cells may provide a personalized treatment platform for the patient's own cells treatment.Objective This study was to evaluate the feasibility of human fibroblasts differentiate toward iPSCs from retinitis pigmentosa (RP) patients and toward iPSC-RPE cells from non-RP individual by retroviral transfection of Oct4,Sox2,c-Myc and KLF4 genes.Methods Human thigh skin tissues were obtained from a RP patient with hotspot mutation of SNRNP200 p.S1087L and individual without SNRNP200 p.S1087L mutation,respectively,with the size 2 c m×3 cm.Human dermal fibroblasts were isolated and cultured by trypsin digestion and explant method.The fibroblasts were transfected by a series of retrovirus and cultured by human embyonic cellconditioned medium to generate and induce iPSCs,and then the iPSCs were identified by morphology,alkaline phosphatase (AP) staining and immunofluorescence assay of specific markers of pluripotent stem cells.iPSCs suspension were injected into SCID mouse to observe the tumorigenesis.The iPSCs from non-RP subject were induced to differentiate toward iPS-RPE cells by embryonic body (EB) inducing method,and iPS-RPE cells were identified by detecting the expression of RPE65,zonula occludens protein 1 (ZO-1) and lecithin retinol acyltransferase (LRAT).Results Cultured human dermal fibroblasts showed fusiform or polygon shape and intercellular close arrangement,and Vimentin was positively expressed in the cells.Small cell colonies were harvested 5-7 days after infected by retroviruses,and the morphology changed from spindle into round mass.The hESC-like iPSCs clonies appeared 20 days after cultivation,and the positive expressions of hESC-specific surface antigens including SSEA3,SSEA4,TRA-1-60,TRA-1-81 and Nanog were found in the cells 25-30 days after cultivation,and the positive staining of AP was obtained 12 weeks after cultivation.A teratoma was formed at the injection site of iPSCs suspension in SCID mouse.Immunofluorescence technique showed that RPE cell-specific proteins including RPE65,ZO-1 and LRAT proteins were positively expressed in iPS-RPE cells at 30 days after differentiation.Conclusions Mutation SNRNP200 p.S1087L of RP patient-specific iPSCs can be induced from human dermal fibroblast by retrovirus infection method.The function and morphology of the iPSCs were similar to hESCs.Human iPSCs cell line generated from the dermal fibroblasts of non-RP individuals can differentiate into iPS-RPE cells.

Objective To explore the clinical characteristics of monochorionic or dichorionic twin pregnancy and the high-risk factors for selective intrauterine growth restriction.Methods 460 women with twin pregnancy were divided into a monochorionic group and a dichorion group.The related clinical features were compared between the two groups.Logistic regression was used to analyze the high-risk factors for selective intrauterine growth restriction.Results The maternal age,conception way,and mode of delivery differed significantly between the two groups (P ＜ 0.05).There were significant differences in the rates of selective intrauterine growth restriction and preterm premature rupture of membranes (P ＜ 0.05).The neonatal weight (large or small) and the rate of neonatal transfer differed significantly (P ＜ 0.05).Logistic regression showed that gestational age and birth weight were the risk factors.Conclusions The chorionic nature plays an important role in the process and outcomes of maternal pregnancy.Monochorionic pregnancy is a high risk factor for selective intrauterine growth restriction,meaning the major cause of selective intrauterine growth restriction may originate from the placenta,with should be a placenta-derived disease.

Objective To evaluate the correlation between diffusion tensor imaging(DTI)measurements,fiber tracking(FT)and the clinical symptoms in patients with cervical spondylotic myelopathy.Methods According to the Japanese orthopaedics association score(JOA),104 patients with cervical spondylopathy were divided into 4 groups:mild in 31 patients with 13-16 scores,moderate in 27 with 9-12 scores,severe in 25 with 5-8 scores,and serious in 21 with 0-4 scores.According to the lesion signal characters,all patients were divided into 3 groups:Group A with normal signal in both T1 WI and T2WI in 33 patients,Group B with normal signal in T1WI but high signal in T2WI in 30 patients,and Group C with low signal in T1 WI and high signal in T2WI in 41 patients.Apparent diffusion coefficient (ADC),fractional anisotropy(FA),λ1,λ2,λ3 were measured in the spinal cord at the serious pressed section,and fiber tractography was performed.The Spearman correlation analyses was used to correlate each of the DTI measurement with JOA score.Group difference was tested with one-way ANOVA method.Results High quality of DTI was acquired in all patients.The FA values in the mild,moderate,severe,and serious groups were respectively 0.69 ±0.13,0.58 ±0.03,0.46 ±0.08,and 0.37 ±0.11 and significant difference was found in different groups(F =100.59,P ＜ 0.05)and positively correlated with JOA scores (r =0.883,P ＜ 0.05).There was no statistical significance between JOA scores and ADC,λ1,λ2,λ3(r=0.232,0.217,0.113,0.127,P ＞0.05).The FA values in group A,B,and C were respectively 0.67 ±0.33,0.51 ±0.21,0.38 ±0.03,and significant difference was found among different groups(F =50.05,P ＜ 0.05).Decrease of JOA score and high signal in T2 companied with decrease of FA value.Decrease of FA values was found associated with increase of fiber bundle damage.The ADC,λ2,λ3 but not λ1 were significantly different among the JOA groups and the group A,B,and C.Conclusions The FA values are positively correlated with clinical symptoms.Decrease of FA values is found associated with increase of fiber bundle damage.DTI can show the severity and extent of damage of spinal cord in patients with cervical spondylotic myelopathy.

Objective To investigate the mid-and long-term effects of metal-on-metal hip resurfacing arthroplasty (HRA),and to analyze its related factors.Methods A total of 64 patients (81 hips) who underwent metal-on-metal HRA from June 2005 to January 2013 were recruited in the present study.There were 35 males (44 hips) and 29 females (37 hips) with the mean age of 48.26±10.45 years (20-65 years),including 47 unilateral and 17 bilateral HRAs.The cohort consisted of osteoarthritis secondary to the developmental dysplasia of the hip (23 cases,29 hips),necrosis of the femoral head (19 cases,22 hips),osteoarthritis (8 cases,10 hips),rheumatoid arthritis (5 cases,9 hips),ankylosing spondylitis (6 cases,8 hips),pigmented villonodular synovitis (2 cases,2 hips) and Otto's disease (1 case,1 hip).During the follow-up duration,radiographic features,including acetabular inclination angle,stem-femoral shaft angle,component loosening,osteolysis,femoral neck narrowing and heterotopic ossification,were evaluated by hip X-rays in straight and froglike position.The size and type of pseudotumor were assessed by MRI and ultrasonography.Clinical efficacy was evaluated by Harris hip score and the University of California at Los Angeles (UCLA) hip score.Considering revision surgery as the end point,the component survivorship was calculated.Results The mean follow-up was 7.98±2.21 years,ranging from 5.0 to 12.8 years.The mean postoperative Harris hip score (92.01 ±5.69) was higher than the preoperative score (41.93 ±9.09).The mean postoperative UCLA pain,walking,function,activity scores (9.37±0.86,9.14± 1.01,8.77± 1.09,6.47± 1.27,respectively) were improved when compared with the preoperative UCLA scores (3.57± 1.23,5.99± 1.30,5.00± 1.01,3.84± 1.41,respectively).The postoperative flexion,abduction and adduction,medial and lateral rotation of the hip was larger than the preoperative ones.Complications occurred in 10 hips (12.3％,10/81).Seven patients (8 hips) experienced early and intermediate complications,including one intraoperative femoral nerve injury,one deep femoral artery and saphenous nerve injuries during the same surgery,one unexplained pain of hip,one femoral neck fracture,three hips of heterotopic ossification,and one pseudotumour.There were mid-and long-term complications in two hips,including one narrowing of the neck and one pseudotumour which was occurred at 9 years.There was one patient (2 hips) underwent revision surgery twice at 5 months and 9 years.The former cause of revision was femoral neck fracture and the latter one was pseudotumour.The Kaplan-Meier survivorship was 98.8％ at five years,and 95.0％ at ten years.Conclusion Patients who underwent metal-on-metal HRA could obtain good mid and long-term results.Pseudotumour and unexplained pain of the hip are critical factors which can affect the mid-and long-term results and survivorship of metal-on-metal HRA.

Estrogen receptors are steroid receptors, widely distributed in skeletal muscle, liver and other tissues.Currently, there are 5 known estrogen receptors.Different estrogen receptors are distributed in different places and have different functions.By mediating different estrogen receptors, estrogen plays an important role in anti-inflammation, promoting proliferation and inhibiting muscle atrophy.Skeletal muscle is the main tissue in the human body, accounting for about 40% of body weight.Skeletal muscle not only plays the role of exercise and support, but also plays an important role in maintaining the body′s metabolism.In recent years, estrogen receptors have received extensive attention in skeletal muscle diseases.Estrogen receptors are considered as potential targets for the treatment of skeletal muscle diseases such as Duchenne muscular dystrophy(DMD)and myotubular myopathy(MTM). This article reviews the research progress of estrogen receptors in skeletal muscle diseases.

Objective: To explore and validate the potential targets of Paeoniae Radix Alba (P. Radix, Bai Shao) in protecting against chemical liver injury through network pharmacology, molecular docking technology, and in vitro cell experiments. Methods: Network pharmacology was used to identify the common potential targets of P. Radix and chemical liver injury. Molecular docking was used to fit the components, which were subsequently verified in vitro. A cell model of hepatic fibrosis was established by activating hepatic stellate cell (HSC)-LX2 cells with 10 ng/mL transforming growth factor-β1. The cells were exposed to different concentrations of total glucosides of paeony (TGP), the active substance of P. Radix, and then evaluated using the cell counting kit-8 assay, enzyme-linked immunosorbent assay, and western blot. Results: Analysis through network pharmacology revealed 13 key compounds of P. Radix, and the potential targets for preventing chemical liver injury were IL-6, AKT serine/threonine kinase 1, jun proto-oncogene, heat shock protein 90 alpha family class A member 1 (HSP90AA1), peroxisome proliferator activated receptor gamma (PPARG), PTGS2, and CASP3. Gene Ontology (GO) enrichment analysis indicated the involvement of response to drugs, membrane rafts, and peptide binding. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis revealed that the main pathways involved lipid and atherosclerosis and chemical carcinogenesis-receptor activation. Paeoniflorin and albiflorin exhibited strong affinity for HSP90AA1, PTGS2, PPARG, and CASP3. Different concentrations of TGP can inhibit the expression of COL-Ⅰ, COL-Ⅲ, IL-6, TNF-α, IL-1β, HSP-90α, and PTGS2 while increasing the expression of PPAR-γ and CASP3 in activated HSC-LX2 cells. Conclusion P. Radix primarily can regulate targets such as HSP90AA1, PTGS2, PPARG, CASP3. TGP, the main active compound of P. Radix, protects against chemical liver injury by reducing the inflammatory response, activating apoptotic proteins, and promoting the apoptosis of activated HSCs.

Objective To investigate the effect of donepezil on subventricular zone ( SVZ) neuro-genesis related neurotrophic factors after cerebral infarction. Methods Mice were randomly assigned into three groups: vehicle-treated sham group (Sham+vehicle,n=18),vehicle-treated middle cerebral artery oc-clusion (MCAO) group (MCAO + vehicle,n=30) and donepezil-treated MCAO group (MCAO+donepezil, n=30). Middle cerebral artery occlusion( MCAO) was induced by thread-occlusion method. Nissl staining was used to measure the infarct volume and the modified neurological severity score(mNSS) was used to as-sess neurologic function and brain water content was detected to assess brain edema degree. Proliferative cells and neuroblasts were labeled with 5-bromodeoxyuridine ( BrdU) and doublecortin ( DCX). The SVZ BrdU+/DCX+cells were detected by immunofluorescence. The expression of glial cell line-derived neurotro-phic factor (GDNF),brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF) were detec-ted by Western blot. Results The infarct volume of MCAO + donepezil group ((13. 33±4. 55)%) was sig-nificantly lower than that of MCAO + vehicle group ((31. 33±3. 93)%,t=7. 34,P<0. 05). The neurologic deficits were significantly ameliorated after donepezil treatment,and the brain water content of MCAO + done-pezil group ((71. 82±10. 18)%)was significantly less than that of MCAO + vehicle group ((85. 93± 7. 54)%,F=13. 480,P<0. 05). All differences were statistically significant (P<0. 05). The area of BrdU+/DCX+cells within SVZ of MCAO + vehicle group ((6. 16±1. 79)%) was significantly larger than that of sham + vehicle group ((2. 25±1. 09)%),and was fewer than that of MCAO+donepezil group ((16. 19± 2. 16)%,F=102. 756,P<0. 05). MCAO significantly promoted the expression of GDNF,BDNF and NGF within SVZ compared with sham operation,and donepezil increased these protein levels(F=15. 114,27. 121, 27. 398,P<0. 05). Conclusion Donepezil regulates neurogenesis via increasesing the expression of GDNF, BDNF and NGF within SVZ after cerebral infarction.