BACKGROUND:Bone marrow mesenchymal stem cells have potential to self-renewal and multi-lineage differentiation. But after a long period of culture in vitro, the proliferation and differentiation capacities of bone marrow mesenchymal stem cells gradual y loss, the mechanism underlying which is not clear now.
OBJECTIVE:To observe the expression of autophagy-related gene Beclin-1 in differentiation from human bone marrow mesenchymal stem cells into neuron-like cells in vitro.
METHODS:The changes of morphological characteristics of neuron-like cells differentiated from human bone marrow mesenchymal stem cells induced by epidermal growth factor were observed. The expression of neuron-specific enolase and glial fibril ary acidic protein in treated and untreated human bone marrow mesenchymal stem cells were detected using immunocytochemistry. The Beclin-1 protein expressions were detected by western blot before and after induction.
RESULTS AND CONCLUSION:After being induced, human bone marrow mesenchymal stem cells presented classical neuron-like morphology;the expressions of neuron-specific enolase and glial fibril ary acidic protein were 78.7%and 8.1%, respectively. The expression of Beclin-1 protein was changed correspondingly during the induction, which increased after 30 minutes of induction and decreased gradual y after 1 hour of induction. Human bone marrow mesenchymal stem cells could be induced into neuron-like cells in vitro by epidermal growth factor. Autophagy-related gene was highly expressed in the induction of early differentiation and the expression gradual y reduced until it remained at a low level during the differentiation.

Objective To observe and analyze the clinical efficacy of docetaxel and S-1 in treatment of anthracycline-resistant triple-negtive breast cancer(TNBC).Methods 64 cases with TNBC admitted in People' Hospital of Shaoxing City from June 2009 to June 2011were selected as research object.The clinical data of these cases were analyzed retrospectively.Anthracycline had been used to treat the cases, but with no effect or recurrence.Then docetaxel combined with S-1 was applied to the cases.Clinical efficacy and adverse reactions of the chemotherapy were observed and analyzed. Results After treatment,the efficiency of 64 cases was 54.69%,and the disease control rate was 79.69%.During the treatment,the main adverse reactions were gastrointestinal reactions and bone marrow suppression,without death,and patients could tolerate the adverse event.Follow-up results showed that the median time of progression was 10.5 months.After two years,progression-free survival rate was 0.Conclusion Docetaxel combined with S-1 has good clinical efficacy for anthracycline-resistant triple-negtive breast cancer,with relatively mild side effects,which may be an ideal adjuvant chemotherapy method.

Objective:To compare the therapeutic effects between Shensongyangxin capsule combined metoprolol and metoprolol monotherapy in treating ventricular premature beats.Methods:The related literature of Shensongyangxin capsule combined metoprolol in treatment of ventricular premature beats published from Aug 2005 to Aug 2013 were searched by computer from Cochrane Library,MEDLINE,EMbase,Wangfang Data,China National Knowledge Infrastructure (CNKI)and Chinese biomedical literature database (CBM),and were searched manually.RevMan 5.0 software was used to perform Meta analysis on data.Results:Symptomatic relief in treatment group was signifi-cantly better than that of control group [(Z=8.28,P <0.00001;OR=6.17,95%CI (4.01,9.48)];dynamic ECG improvement in treatment group was significantly better than that of control group [Z=9.37,P <0.00001;OR=3.41,95%CI (2.64,4.40)];there was no significant difference in incidence rate of adverse drug reaction be-tween two groups [Z=1.47,P =0.14;OR=0.70,95%CI (0.43,1.13)].All funnel schemas were symmetry in general,so there was small possibility of bias.Conclusion:Shensongyangxin capsule combined metoprolol possesses better therapeutic effect than metoprolol monotherapy in treating ventricular premature beats,and it doesn′t in-crease adverse drug reactions.

Objective To investigate the changes of T cells and dendritic cells (DCs) and their related factors in children with immune thrombocytopenia (ITP) before and after therapy,and to analyze their clinical significance.Methods T-cells and DCs subsets were determined by flow cytometry both in 64 children with ITP (ITP group) before and after therapy and the control group.The serum levels of interleukin (IL)-4,interferon-γ (IFN-γ),transforming growth factor beta 1 (TGF-β1),and IL-27 were detected by enzyme linked immunosorbent assay (ELISA).Results Treatments of glucocorticoid or IVIg were effective in 41 cases of 64 ITP children.Compared to the control group,helper T cells (Th),Th/suppressor T cells (Ts),T regulatory cells (Treg),plasmacytoid DC (pDC),pDC/myeloid DC (mDC),and TGF-β1 in ITP patient group before treatment were significantly lower,while IFN-γ and IFN-γ/IL-4 were significantly higher (P ＜0.05).In ITP group,Th,Th/Ts,Treg,pDC,pDC/mDC,TGF-β1,and IL-27 were significantly increased,while IFN-γ and IFN-γ/IL-4 were decreased in children with ITP after therapy and achieved response (P ＜ 0.05).However,there was no significant difference between before and after therapy in ITP children without treatment response (P ＞ 0.05).Conclusions T cells and DCs subsets disorder and abnormal cytokine levels are observed in children with ITP,which can be corrected by immunosuppressive therapy,indicating that Th1 overactivity and the decrease of Treg and pDC both in quantity and function may be related to the pathogenesis of ITP in children.

Objective To validate the clinical applicability of the published standard reference interval of routine clinical chemistry(WS/T 404.1-2012,WS/T 404.1-2012) based on the results of health examination.Methods This was a retrospective study.The results of serum TP(Biuret),ALB (BCG),ALT(Rate without 5'-pyridoxal phosphate),AST (Rate without 5'-pyridoxal phosphate),ALP (Rate,AMP),GGT(Rate) from healthy examinatiou individuals (from January to July,2013) were collected to calculate the 2.5％ and 97.5％ pereentiles,excluding the significant abnormal results according to the Medical Deciding Level 2 recommended by Staland.The number of the cases after excluding were 19 708(M 12 044,F 7 664) 、19 728(M 12 069,F 7 659) 、45 569(M 26 299,F 19 270) 、45 877(M 26 739,F 19 138)、5 965(M 4 208,F 1 757)、4 726(M 3 164,F 1 562),respectively.The proportions of all the examined results (with and without the excluded results) that fell in the published standard reference interval were also calculated.Results The test of normality revealed that the frequency distributions of all verified items were skewed distributions after excluded the abnormal results.The 2.5％-97.5％ percentiles of TP,ALB,ALT(M),ALT(F),AST (M),AST (F),ALP (F20-49),ALP (F50-79),GGT (M) and GGT (F) were 64-79 g/L,40-59 g/L,9-52 U/L,7-39 U/L,13-41 U/L,12-33 U/L,42-116 U/L,36-98 U/L,44-130 U/L,11-72 U/L and 7-50 U/L respectively.More than 90％ results of TP,ALB,ALT(M),ALT(F),AST (M),AST(F),ALP(F20-49),ALP(F50-79),GGT(M) and GGT(F) (with and without the excluded results) fell in the reference intervals of national standards,GGT(M) was 80％ and 91％.Conclusions The published standard reference interval of routine clinical chemistry (WS/T 404.1-2012,WS/T 404.1-2012) are applicable to our laboratory.

Objective To study the glutamine combined with early enteral nutrition’s effects on severe acute pancreatitis’systemic inlfammation. Methods 150 Cases with severe acute pancreatitis were divided into total parenteral nutrtion group(TPN group), early enteral nutrition group(EEN group) and Glutamine and early enteral nutrition group(G+EEN group) according to their therapeutic methods. The curative effects, APACHEⅡscore and liver and kidney function were compared after treatment. Inlfammatory cytokines of hs-CRP, TNF-α, IL-1β, IL-6, IL-8 and IL-10 before and after treatment were detected and compared. Results The efifciency rate in G+EEN group was signiifcantly better than that in TPN and EEN group(P<0.05), with lower APACHEⅡscore and better liver and kidney function. The level of hs-CRP, TNF-α, IL-1β, II-6 and IL-8 after treatment in G+EEN group were signiifcantly lower than that in TPN and EEN group(P<0.05), except IL-10, which was signiifcantly higher than that in TPN and EEN group (P<0.05). Conclusion Glutamine combined with early enteral nutrition could signiifcantly ameliorate severe acute pancreatitis’systemic inlfammation, its curative effects is better than early enteral nutrition and total parenteral nutrition.

Objective To evaluate color Doppler ultrasonography(CDU) in the diagnosis of intraocular tumor. Methods Two hundred seventy-three cases(301 eye) of intraocular space-occupying lesions were examinated by high-resolution CDU to observe the two-dimensional gray-scale image and vasculature of intraocular tumors. Results Intratumor calcifying spot could be detected in 138 cases(149 eye) of 153 cases(178 eye) with retinoblastoma,and intratumor blood flow with branchy pattern could be found in 152(177 eye) of 153 cases(178 eye) with retinoblastoma. In 4 Coats disease and 2 persistent hyperplastic primary vitreous and 3 retinopathy of prematurity, intratumor calcifying spot couldn′t be detected,and the band blood flow of the detached retinal vessels could be detected. Intratumor blood flow with branchy pattern could be detected in all 66 choroidal melanomas and 4 melanomas of ciliary body with infiltrated choroid and 15 of 19 choroidal metastases. Intratumor blood flow with “simulating vessel pool” or conglomerate sparkled pattern could be found in 12 choroidal hemangioma. No intrinsic flow was detected in 8 choroidal hemorrhages or subretinal hemorrhages and 2 teratoid medulloepitheliomas. Conclusions CDU is of significance in the diagnosis and differention of intraocular tumors.

The up-regulation mechanism of membrane type I matrix metalloproteinase (MT1-MMP) in macrophages stimulated by silica in vitro and the contribution of early growth response 1 (Egr-1) transcription factor in the gene expression pathway were investigated. Macrophages stimulated by silica were treated with Egr-1 antibody or Egr-1 decoy oligodeoxynucleotides (ODN). The levels of MT1-MMP proteins were determined by Western blot and the expression of MT1-MMP mRNAs was detected by RT-PCR. The results showed as compared with control macrophages, silica-stimulated group showed up-regulated gene expression of MT1-MMP via Egr-1 (P<0.01). Compared with silica-stimulated macrophages untreated with antibody, the cells treated with 5 microg/mL Egr-1 antibody were associated with reduced expression of MT1-MMP protein (P<0.01) and mRNA (P<0.01). Compared with silica-stimulated untransfected group, the Egr-1 "decoy" ODN group was associated with reduction in the expression of MT1-MMP protein and mRNA (P<0.01). It was concluded gene expression of MT1-MMP which may play a critical role in silicosis was up-regulated by silica in macrophages. Egr-1 participated in the expression of MT1-MMP and positively regulated the expression. Both Egr-1 antibody and Egr-1 decoy ODN suppressed the expression of MT1-MMP through the Egr-1 pathway and may become a potential therapeutic tool in the management of silicosis in the future.

Objective To compare disk diffusion with E-test methods for clarithromycin susceptibility testing of Helicobacter pylori (H.pylori).Methods A total of 44 strains of H.pylori were isolated from gastric mucosa biopsy from patients undergoing gastroscopic examination.Disk diffusion and E-test methods were used for clarithromycin susceptibility testing of H.pylori.The agreement of disk diffusion and E-test was assessed by linear regression analysis.Results The minimum inhibitory concentration (MIC) tested by E-test method ranged from 0.016 to 256 μg/mL,and drug resistance was observed in 12(27.3％) isolates.In range of 0-35 mm of inhibition diameter,the results of disk diffusion method were correlated well with the MICs obtained by E-test method (r2 =0.91,P ＜0.01).Regression analysis showed that with inhibition diameters≥ 18 mm as considered sensitive to clarithromycin and ≤ 15 mm as resistant,the agreement was 100％ between two methods.Conclusion The disk diffusion method is equivalent to the E-test method for clarithromycin susceptibility testing of H.pylori,which can be an alternative method for clinical application.

Recently, research on collagen attracts more interests due to its good biological compatibility. The present study attempted to establish a fast and efficient method to purify collagen from soft-shelled turtle and to explore its application in biological materials. The structure and type of collagen fiber in calipash were determined by van Gieson staining and Picrosirius red staining, which could contribute to the isolation of collagen from soft-shelled turtle Calipash (STCC). Collagen fibers were in high content and the main collagen fiber was type I in STCC. The crude STCC solution was purified by dialysis with different cut-off molecular weight. SDS-PAGE demonstrated that the best purification was in applying 100 kDa dialysis bags after 48 h. The water absorbing capacity and holding capacity of STCC were up to 12.06 g/g and 98.21%, respectively. STCC can be degraded by collagenase in vitro entirely after 72 h. The hemolysis, skin sensitization, hemostatic and wound healing of STCC were determined by using SD rat model, and the collagen cross-linked by glutaric dialdehyde was set as a comparison. STCC and STCC cross-linked did not result in destructed red blood cell, inflamed and sensitized skin. Both materials exhibited good hemostatic effect. Thus, STCC improved the wound healing efficiently. This study implies a potential of STCC in the field of biomaterial.
Animals ; Biocompatible Materials ; Biotechnology ; Collagen ; isolation & purification ; Electrophoresis, Polyacrylamide Gel ; Molecular Weight ; Rats ; Rats, Sprague-Dawley ; Skin ; drug effects ; Turtles ; Wound Healing