The establishment of induced pluripotent stem cells(iPSCs)has been a major breakthrough in the field of stem cell research since 2006,and it made possible for the use of stem cells in treating retinal degenerative diseases.Research showed that fibroblast,B lymphocytes,neural stem cells,hair corneous cells,pancreatic cells,mesenchymal cells of umbilical cord stroma and amniotic membrane can be reprogrammed as iPSCs,and they are capable of differentiating into specific types of cells.Some novel developments in iPSCs study in ophthalmology also were observed over the past few years.Induced iPSCs can differentiate into retinal pigment epithelial cells,photoreceptors and other retinal cells,which lay a foundation for the therapy of retinal degenerative diseases.Differented from traditional treatment of stem cells,the generation of iPSCs makes it possible to utilize somatic cells derived from patients for stem cell therapy without provoking ethical and immunological problems.The generation of iPSCs,the current research about iPSCs in the ophthalmic field,the limitations of iPSCs in the clinic and their future development and application were reviewed.

AIM: To examine the autoantibody against ? 1-adrenoceptor and its biologic activities during the development of renal hypertension. METHODS: Renal hypertension of rat was achieved by clipped renal artery, the titre of autoantibody to ? 1-adrenoceptor was detected using ELISA immunoassay. Furthermore, the biological offects of these autoantibodies on cultured cardiomyocytes were also examined. RESULTS: After two weeks of clipping renal arteries, both the frequency of occurrence and the titre of autoantibodies to cardiac ? 1-adrenergic receptor were significantly increased as compared with the control of pre-treatment. The increased autoantibodies lasted for several weeks and then automatically decreased gradually to the pre-clipping level at 12 weeks. The biological effects of these autoantibodies displayed an "agonistic-like" activities on the beating frequency of cultured neonatal cardiomyocytes. CONCLUSION: Autoantibodies against ? 1-adrenoceptor may play a role in the elevation of peripheral vascular resistance and in the development of cardiac hypertrophy in rats with renal hypertension.

Objective To construct the eukaryotic vector that expressing hepatitis B virus (HBV) S and the single fragment of variety chain (ScFv) of monoclonal antiboy against cytotoxic Tlymphocyte-associated antigen-4 (CTLA-4) and to analyze the immunological activity of recombinant S-ScFv protein.Methods The oringially constructed pSect2/ScFv4F10 and pSect2/S were double enzyme digested by Sfi I and Hind Ⅲ,respectively.Then the HBV S gene was cloned into the pSect2/ScFv4F10 vector.The pSect2/ScFv4F10 and pSect2/S-ScFv4F10 were expressed in Chinese hamster ovary (CHO) cells,and the expressed proteins were verified through sodium dodecyl sulfatepolyacrylamide gelelectrophoresis(SDS-PAGE)andWesternblotting.Afterultrafiltration concentration and affinity chromatography,the biological affinity of the expressed ScFv4F10 and S-ScFv4F10 proteins were examined by competitive enzyme-linked immunosorbent assay (ELISA) and surface plasmon resonance (SPR) technology.The comparison between groups was done by One-way ANOVA.ResultsThe eukaryotic expression vector of pSect2/S-ScFv4F10was successfully constructed,and relative molecular mass of the expressed protein of S-ScFv4 F10 was about 52 000 that analyzed by SDS-PAGE and Western blotting.With the fixed concentration of 4F10-mAb against CTLA-4,the A570 value of the mixed reaction with purified CTLA-4 antigen gradually increased with the decrease of ScFc fusion protein proportion; when the molar ratio of ScFv,S-ScFv4F10∶4F10=2∶1,the competitiveinhibitionratesagainst 4F10conjugatedantigenwere72.6％and64.5％,respectively.The affinity constants of association kinetics for CTLA-4 mAb,ScFv4F10 and S-ScFv4F10 with CTLA-4 antigen were 7.29 × 108 mol/L,9.52 × 106 mol/L and 2.04 × 106 mol/L,respectively,and the dissociation constants of KD were 1.40 × 10-9 mol/L,1.05 × 10-7 mol/L and 4.91 × 10-7 mol/L,respectively.ConclusionsThe eukaryotic expression vector of pSect2/S-ScFv4F10is successfully constructed,and the recombinant protein of S-ScFv4 F10 has a fairly high affinity with CTLA-4 antigen.

Animals ; Antineoplastic Agents ; therapeutic use ; Bone Density Conservation Agents ; therapeutic use ; Bone Neoplasms ; drug therapy ; secondary ; Breast Neoplasms ; drug therapy ; pathology ; Chemotherapy, Adjuvant ; Diphosphonates ; therapeutic use ; Female ; Humans ; Imidazoles ; therapeutic use ; Neoadjuvant Therapy ; Osteoporosis ; prevention & control

The valuators affect the results of performance evaluation directly.By construing the objective requirements of performance evaluation,the motivation of stakeholders,and the diversity of evaluating objects,we then set forth the necessity of valuator's multipluralism in the performance evaluation of comprehensive prevention and treatment of HIV/AIDS.We also put forward another pertinent advice that multiplural valuators should take part in general HIV/AIDS prevention and treatment evaluation,thus providing a reference for guaranteeing the results of performance evaluation,and the advancement and improvement in the performance of HIV/AIDS comprehensive prevention and treatment.

BACKGROUND:To construct tissue engineering cartilage would open up a novel way for the repair of cartilage damage in avoidance of the disadvantages of traditional therapeutic method.OBJECTIVE: To probe the techniques for the isolation of mesenchymal stem cells (MSCs) from bone marrow, as well as the in vitro differentiation into chondrocytic phenotype in a specific culture fluid.DESIGN:A complete randomized experimentSETTING:The Department of Traumatic Orthopedics and Hand Surgery of the First Affiliated Hospital of Guangxi Medical University, and Teaching and Research Faculty of Histology and Embryology of Guangxi Medical University.METHODS: The experiment was carried out at Guangxi Medical University between August 2002 and April 2003. Twenty SD neonatal weaning rats were selected. Bone marrow was aspirated from the bones of rat limbs and was isolated by gradient centrifugation in Percoll, and MSCs could be obtained in combination with adherent screening method, which were then cultured in DMEM-LG with 15% fatal bovine serum (FBS) in the incubator of 37℃ with 5% CO2 for 10-14 days. The passage cells were induced in DMEM-HG with 15% FBS (containing TGF-β1 10 μg/L, 10-7 mol/L dexamethasone, 50 mg/L VitC).MAIN OUTCOME MEASURES :The morphology, growth, as well as proliferation and specific expression of chondrogenic matrix of in vitro cultured MSCs due to specific induction.RESULTS: Totally 20 SD rats were observed and analyzed with no loss SCs grew in visible symmetric colonies, displaying a long-spindle shape,and the morphological characteristics of marrow-derived MSCs had no obvious changes during passage-culture, but its proliferation time was found from a shuttle fibroblastic appearance to polygonal shape, displaying posiHC staining of type Ⅱ collagen of cartilage specific matrix.bronectin adherent screening technique is a convenient, effective and practical method to separate and collect MSCs from rat bone marrows in chondrogenic phenotype when induced by a specific medium and can secrete cartilage specific matrix, and they can be the optimal seed cells for cartilage tissue engineering.

Objective To describe the clinical and muscle pathological characters of juvenile myositis patients with perifascicular atrophy (PFA).Methods A series of 21 consecutive muscle biopsies with clinically and pathologically confirmed juvenile myositis were studied.All biopsies had PFA of muscle fibers.Results Clinical manifestation:11/21 had typical dermatomyositis rash,9/21 possible dermatomyositis because of atypical rash,1/21 mixed connective tissue disease;9/21 had weakness complaint;9/21with normal creatine kinase (CK) level,4/21 low grade rise,8/21 apparently elevated,10/ 11 definite dermatomyositis patients with normal or low grade level;6/21 combined with interstitial lung disease,2 with EB virus infection,1 with cytomegalo virus infection.Muscle pathology:4 specimens with atypical PFA also had enzyme abnormality in nicotinamide adenine dinucleotide,succinate dehydrogenase and cytochrome c oxidase staining,9/12 cases had membrane attack complement deposition intramuscular capillaries,and they had prominent regional mitochondrial abnormalities,protrude PFA and focal muscle damage.Skin biopsy of one case showed perivasculitis in dermis.Conclusions PFA could be seen in juvenile dermatomyositis and mixed connective tissue disease,and PFA could not increase the CK level.The capillary and intermediate-sized vessels damage may cause chronic ischemia,which could explain the PFA with mitochondrial abnormalities.

CAS (Chrome azurol S) assay was a universal method for detecting the bacterial siderophores, and Sugar-Asn liquid medium has been applied to the studies on siderophores from Pieudomonas. In this paper, Asp has been substituted for Asn, and MSA-CAS agar plate was developed by integrating the MSA (sugar-Asp) medium and CAS bright blue dye, which has been used in the universal CAS assay. On the aspect of siderophores detection , 8 strains of 7 species from Pseudomonas had been screened on MSA-CAS agar plates and universal CAS assay respectively. The results showed that MSA-CAS agar was higher-sensitive and lower basic fluorescent than universal CAS assay.

Objective The purpose of this investigation was to investigate the neuroprotective effects of rhubarb for ICH,as well as its mechanism.Methods ICH was produced in adult Spargue-Dawley rats by injection of collagenase IV(0.05U/0.5uL).Intraperitoneal injection of rhubarb (70mg/kg) or saline,was started at 3,6 or 12 hours post-ICH respectivdy.Casepase-3 activity.TUNEL and neurological behavior function were performed 24hours after ICH.Results Rhubarb siven at 3 or 6 hours can inhibit casepase-3 activity(P<0.001),reduce TUNEL positive cells(P<0.05) and attenuate apomorphine-induced rotation(P<0.05) at 24 hours after ICH.However,the animals which were treated 12 hours showed no improvement.Conclusion Rhubarb may be a potential drug for ICH patients for its possible effect of inhibiting apoptosis.

Objective:Through detecting miRNA-26a,β-catenin,GSK-3β and α-SMA expressions in IgA nephropathy with varying degrees of renal interstitial fibrosis,the study was performed to explore the effect of miRNA-26a targeting GSK-3β on Wnt/β-catenin signal pathway simulated renal interstitial fibrosis.Methods:Incorporated 46 cases of IgA nephropathy patients were divided into three group based on the degree of renal interstitial fibrosis,namely,mild group,moderate group and severe group;7 cases of normal renal tissues away from the renal tumor tissues were selected as the control group.Expression levels of miRNA-26a in renal tissues of each group were detected based on RT-qPCR method,to analyze the correlation between miRNA-26a and renal fibrosis In patients with IgA nephropathy.Furthermore,mRNA and protein expression levels of β-catenin,GSK-3β and α-SMA in renal tissues were measured using RT-qPCR and immunohistochemistry,respectively,the comparison was then made in each group;subsequently,correlation analysis was further conducted to investigate the relationship of miRNA-26a with β-catenin,GSK-3β and α-SMA.Results:(1) Compared with the control group,miRNA-26a expression was down-regulated from renal biopsy of IgA nephropathy patients,the expression level of miRNA-26a was significantly decreased,showing statistical differences among groups (P＜0.05).(2) Compared with the control group,mRNA and protein expression levels of β-catenin,GSK-3β and α-SMA in renal tissues were all increased in IgA nephropathy patients,and the degree of expression increased gradually with the increase of the degree of renal interstitial lesion,differences were statistically significant among groups (P＜0.05).(3) Correlation analysis results indicated that there were negative correlation between miRNA-26a expression in renal tissues and the degree of renal interstitial fibrosis,differences were statistically significant among groups (r=-0.943,P ＜0.05),at the same time,expression intensities of GSK-3β,β-catenin and α-SMA in renal interstitium and renal tubules were positively correlated with the degree of renal interstitial fibrosis(r =0.917,P＜0.05;r =0.943,P＜0.05;r =0.926,P ＜0.05),meanwhile,positive correlation was also found regarding protein expression of GSK-3β and β-catenin in renal interstitium (r=0.834,P＜0.05).Conclusion:Collectively,miRNA-26a can be involved in Wnt/β-catenin signal pathway simulated renal interstitial fibrosis via the regulation of GSK-3β.