Myeloid-derived suppressor cells ( MDSC) are a bone marrow-derived heterogeneous cell population with immuno-suppressive activity.Although there is convincing evidence that autoimmune diseases are associated with MDSC expansion ,controversies remained regarding the role of MDSCs in controlling autoimmune responses .Recent studies have shown that the expansion of MDSCs , which are capable of inhibiting effector cell function in vitro ,does not always lead to alleviation of autoimmune diseases ,and in some ca-ses paradoxically exacerbates the disease progression .This review summarizes recent insights into the role of MDSCs in the development of autoimmune responses and the potential of using MDSCs for the treatment of autoimmune diseases .

Objective To study the effects of cleavage of Bcl-2 by two DNAzymes on apoptosis of human hepatoma cell line (HepZ1).Methods Two “10-23” DNAzymes(DzT and DzTi) targeting Bcl-2 mRNA and their analogues(DzT' and DzTi') were synthesized and used to cleave Bcl-2 mRNA in vitro and in BEL-7402 cells.The RT-PCR was performed to assess the cleaving efficiency.Expression of Bcl-2 protein was determined by immunofluorescent method.Cell apoptosis was detected by flow cytometry.Results The unmodified Enzymes DzT,and its modified form DzTi,which had an added 3'-inverted thymidine,could effectively cleave Bcl-2 mRNA in vitro.After transfected into BEL-7402 cells,DzTi exhibited more powerful cleaving ability than DzT,significantly down-regulated the level of Bcl-2 protein(P ＜0.01) and inhibited the cell growth(P ＜0.05).The results of flow cytometry suggested that the apoptosis rate of DzT and DzTi significantly increased,appeared apoptotic peak.Cell cycle was delayed in DzT and DzTi group,proportion of cells in G0/G1 increased,S phase cells decreased.Conclusion The synthesized DNAzymes could effectively cleave Bcl-2 mRNA,decrease the level of Bcl-2 protein and induce hepatoma cells apoptosis.

Objective To study the inhibitory effect of hammerhead ribozyme targeting connective tissue growth factor(CTGF) on collagen I synthesis and cell cycle progression of human hepatic stellate cell line(LX-2) cells.Methods Hammerhead ribozyme cDNA targeting CTGF mRNA plus two self-cleaving sequences were inserted into pTriEx2 vector to construct a recombinant vector pTriCTGF-Rz.LX-2 cells were transfected with either pTriEx2 or pTriCTGF-Rz and further stimulated with or without TGF-1.There were five groups in the experiment:control group,pTriEx2 group,pTriCTGF-Rz group,pTriEx2 plus TGF-?1 group,and pTrCTGF-Rz plus TGF?1 group.Semi-quantitative RT-PCR was used to detect the levels of CTGF mRNA and collagen Ⅰ mRNA.ELISA and flow cytometry were used to detect the levels of collagen Ⅰ secretion and cell cycle.Results Transfection of pTriCTGF-Rz into LX-2 cells reduced the CTGF mRNA and collagen Ⅰ mRNA levels as well as collagen Ⅰ protein level compared with pTriEx2 group(P

Objective:To observe influence of cardiac resynchronization therapy (CRT ) on ventricular remodeling and inflammation in patients with chronic heart failure (CHF) .Methods :A total of 84 CHF patients treated in our hospital from Jun 2012 to Feb 2015 were selected , according to randam number table , they were randomly and e‐qually divided into routine treatment group (received routine medication ) and combined treatment group (received CRT based on routine treatment group) .Left ventricular ejection fraction (LVEF) ,left ventricular end -diastolic dimension (LVEDd) ,6min walking distance (6MWD) ,levels of high sensitive C reactive protein (hsCRP) ,inter‐leukin (IL)‐6 and tumor necrosis factor (TNF)–αwere compared between two groups before and three months after treatment .Results:Compared with before treatment ,three months after treatment ,there were significant rise in LVEF and 6MWD ,and significant reductions in LVEDd ,levels of hsCRP ,IL‐6 and TNF‐α in combined treatment group ( P<0.05 or < 0.01 ) ,while there were no significant improvements in above indexes in routine treatment group .Compared with routine treatment group ,there were significant rise in LVEF ± [(29.42 ± 4.32)%vs .(37.16 ± 4.72)% ] and 6MWD [ (232.66 ± 40.54) m vs .(304.12 ± 51.65) m] ,and significant reductions in LVEDd [ (64.35 ± 7.81) mm vs .(57.64 ± 6.12) mm] ,levels of hsCRP [ (23.21 ± 3.45)μg/ml vs .(16.31 ± 2.02)μg/ml] ,IL‐6 [ (22.08 ± 3.82)μg/ml vs .(15.79 ± 2.09)μg/ml] and TNF‐α[ (32.66 ± 5.66)μg/ml vs .(23.23 ± 3.12)μg/ml] in combined treatment group , P<0.05 or <0.01. Conclusion:CRT can significantly reduce levels of hsCRP ,IL‐6 and TNF‐αin CHF patients ,which may be the main mechanism delaying ventricular remodeling and improving cardiac function .

Objective To establish an aging model of mesenchymal stem cells (MSCs) and to investigate aging related biological mechanism for the purpose of studying the senesence of MSCs .Methods MSCs were separated and purified from human placenta, and the cells of the third passage(P3-MSCs) were cultured in the medium for 2 hours, then 100,200 and 300 μmol/L hydrogen peroxide ( H2 O2 ) was added to the cells for 2 hours to establish the MSCs aging model in vitro. Biological characteristics of aging MSCs were evaluated by cell cycle assay and senescence associated β-galactosidase staining.The expression of p16,p21 and p53 genes was further measured using quantitative real-time PCR (RT-PCR).Re-sults Compared with the control , the number of MSCs treated with 200μmol/L H2 O2 for 2 hours was significantly decreased and the cells displayed less adipogenic ,osteogenic and chondrogenic differentiation .Moreover ,after exposure to 200 μmol/L H2 O2 , the majority of the cells were in the G 0/G1 phase as showed by cell cycle analysis .The percentage of senescence-associated β-galactosidase-positive cells was increased , and the expression of p 16 , p21 and p53 mRNA and protein was significantly increased.Conclusion The results of this study has demonstrated that the H 2 O2 (200 μmol/L) can be used to establish the aging model of MSCs in vitro, and the cellular phenotypic alteration may attribute to the cell cycle associated gene expression (p16, p21, and p53).

Objective To investigate the effects of large dose of atorvastatin on the expression of Sprouty-1 in CD4 + T lymphocytes from unstable angina patients during perioperative period of PCI.Methods A total of 52 unstable angina patients enrolled were divided randomly (random number) into large-dose atorvastatin (80 mg/d,n =26) pretreated group and moderate-dose atorvastatin (20 mg/d,n =26) pretreated group.Circulating CD4 + T cells were obtained by magnetic cell sorting system (MACS) before PCI and 18-24h after PCI.For detecting the gene expression,the reverse transcription fluorescent quantitative polymerase chain reaction (RT-PCR) was used to measure the expression of Sprouty-1 mRNA in CD4 + T lymphocyte.The level of Sprouty-1 protein was detected with Western blot analysis and IL-2 was quantified by enzyme-linked immunosorbent assay (ELISA).Results ①Compared with large-dose group before PCI,the expression of Sprouty-1 mRNA and Sprouty-1 protein levels were dramatically increased in large-dose group after PCI (P ＜ 0.05).②Compared with moderate-dose group before PCI,the expression of Sprouty-1 mRNA and protein levels were slightly increased in moderate-dose group after PCI,but there was no statistical significance (P ＞ 0.05).③Compared with large-dose group before PCI,the serum level of IL-2 was decreased in large-dose group after PCI (P ＜ 0.05).Whereas the serum level of IL-2 was slightly increased in moderate-dose group after PCI compared to moderate-dose group before PCI,there was still no statistical significance (P ＞ 0.05).Conclusions Large-dose atorvastatin pretreatment reduced post-PCI myocardial inflammation through up-regulating the expression of Sprouty-1 mRNA and level of Sprouty-1 protein in CD4 + T lymphocytes.

Objective:To construct the eukaryotic expression plasmid carrying hTERT-P2A-EGFP, and to explore its expression and transfection efficiency in the HEK293FT cells.Methods:The recombinant plasmid was constructed by using pBABE-puro-hTERT and pRRLSIN-cPPT-MSCV-EGFP plasmids.The hTERT,P2A,and EGFP genes were obtained using pBABE-puro-hTERT as template by PCR.And the correct hTERT was inserted into pRRLSIN-cPPT-MSCV-EGFP vector.Then the recombinant plasmid containing hTERT-P2A-EGFP gene was obtained and identified.The HEK293FT cells were transfected by the recombinant plasmid, and the expression of green fluorescence protein(GFP) was observed by fluorescence microscope.Results:The PCR results showed that the fragments of hTERT, P2A, and EGFP were 3 400, 110 and 720 bp.And the length of gene fragment(hTERT-P2A-EGFP)was 4 300 bp by enzyme digestion.The results of sequencing showed that the 1 547 site of the target gene was mutated.Using site-directed mutagenesis, the 1 547 site was successfully mutated.And the target gene sequence was completely identical with the sequence published in GenBank.The recombinant plasmid was transfected into the HEK293FT cells, and GFP was observed in the cells.The results of flow cytometry showed that the transfection efficiency of recombinant plasmid was 44.8%.Conclusion:The recombinant plasmid carrying hTERT-P2A-EGFP gene is successfully constructed, and it can be used for cell transfection.

BACKGROUND:Placenta is a valuable source of mesenchymal stem cels for stem cel therapy and future application in the field of regenerative medicine. However, conventional methods cannot acquire a large amount of purified human placenta-derived mesenchymal stem cels. Here, we present a new method for isolating human placenta-derived mesenchymal stem cels suitable for banking strategies and for future clinical applications. OBJECTIVE:To analyze the biological characteristics of human placenta-derived mesenchymal stem cels cultured by tissue dissociating and colagenase digestion. METHODS: Human placenta-derived mesenchymal stem cels were obtained from human placenta by tissue dissociating and colagenase digestion method. Immunophenotype was analyzed by flow cytometry. Growth curve was determined by MTT assay, and differentiation ability was evaluated byin vitro adipogenic, osteogenic and chondrogenic induction as wel. RESULTS AND CONCLUSION:Human placenta-derived mesenchymal stem cels could be passaged stablyin vitro. Furthermore, the cels expressed CD73, CD90, CD105, but were negative for the markers of CD11b, CD19, CD34, CD45, and HLA-DR. Human placenta-derived mesenchymal stem cels proliferated actively and began to grow logarithmicaly at days 3-5 folowed by a plateau period at day 6. In addition, the isolated cels could be induced into adipocytes, osteocytes, chondrocytesin vitro. In a word, the results of this study demonstrated that the tissue dissociating and colagenase digestion method is an efficient method for obtaining a large amount of human placenta-derived mesenchymal stem cels that can be stably cultured in vitro and have strong proliferative ability.

Objective To study the effects of acupuncture combined with Anchang powder acupoint application in the treatment of irritable bowel syndrome with diarrhea. Methods A total of 100 patients with irritable bowel syndrome with diarrhea in our hospital from July 2014 to July 2016 were enrolled.The subjects were randomly divided into the control group (n=50) and the treatment group (n=50). The control group were treated with trimebutine, while the treatment group were treated with acupuncture combined with Anchang powder acupoint application. The two groups were treated for 4 weeks. The clinical effects of the two groups after treatment were compared. The clinical symptom scale was used to evaluate the clinical symptoms, and the quality of life was evaluated by the quality of life scale (QOL) The recurrence rate after half a year heal of the two groups were compared. Results The total efficacy rate of the treatment group was 92.0% (46/50), significantly higher than 68.0% (34/50) of the control group (x2=9.000, P=0.003). After treatment, the clinical symptom integral(1.89 ± 0.95 vs.4.02 ± 1.13,t=10.202)of the treatment group were significantly lower than the control group(P<0.05).After treatment,the quality of life rating scores(34.15 ± 6.75 vs.28.47 ± 5.01,t=4.803) of the treatment group were significantly higher than the control group (P<0.01). Half a year after treatment, the recurrence rate of the treatment group was 24.1% (7/29), significantly lower than 64.7% (11/17) of the control group (x2=7.405, P=0.007). There was no significant difference of the two groups during the treatment. Conclusions Acupuncture combined with Anchang powder acupoint application showed a good efficacy for the patients with irritable bowel syndrome with diarrhea. And the treatment showed the low incidence of adverse reactions and low recurrence rate, can mprove the clinical symptom and quality of life.

Objective: To report a case of peripheral neuropathy secondary to copper deficiency (CD) by long-term decoppering chelation in Wilson′s disease (WD) to enhance understanding of the disease, and to pay more attention to individualized treatment of WD. Methods: A case of WD diagnosed 12 years ago confirmed by gene detection and since then treated with anti-copper agent was diagnosed as CD based peripheral neuropathy and significant neutropenia and followed up for six months, and the clinical manifestations, laboratory examination, electrophysiology, imaging features were summarized. The related literatures were reviewed. Results: A total of 16 cases of WD complicated with CD were reviewed and analyzed, including seven males and nine females aged 13-56 years. All of them were treated with zinc for 1-38 years, and nine cases with peripheral neuropathy. Hematological indicators can be significantly improved and neurological symptoms can be partially alleviated after stopping copper removal treatment. Conclusions Peripheral neuropathy in a WD with treatment-related CD may occur in blind treatment, irregular treatment monitoring and without individualized treatment adjustment. It is necessary to monitor blood routine, copper and zinc metabolism regularly and advocate individualized treatment of WD.