[Objective]To study the possibility of differentiation of rabbit marrow mesenchymal stem cells(MSCs)into endothelial cells in vitro.[Method]BMSCs of rabbits were obtained by using gradient centrifuge method.The adhesive cells were preserved to passage culture to get pure MSCs.MSCs in the second generation were used.After induction with vascular endothelial growth factor(VEGF),microscopic exarm,cytoimmunofluorescent staining confirmd the trial group cells becomes to endothelial cells.W-P corpuscles which was the symbol of endothelial cells were observed by transmission electron microscope.[Result]The differentiated cells demonstrated the characters of endothelial cells under phase contrast microscopy.The cytoimmunofluore-scent staining showed that the trial group cells were induced to endothelial cells.W-P corpuscles could be seen under transmission electron microscope.[Conclusion]The adult rabbit BMSCs can present features of endothelial cells in certain induction medium,they are ideal donor cells of tissue engineering vascularization.

Objective: To prepare a self-designed three-dimensional woven scaffold using poly lactide-co-glycolide acid (PLGA), and to observe the influence of the prepared scaffold on the growth of Schwann cells in vitro and its in vivo degradation. Methods: The 3D scaffolds were prepared by means of melt spinning, extension, weaving, and other procedures. The alignment of micro-tubules was observed under the scanning electronic microscope (SEM). The size of the micropores was also measured. Primary cultured Schwann cells were seeded on the 3D scaffolds, and the growth, adherence, proliferation, and apoptosis of Schwann cells were observed under inverted phase contrast microscopy and SEM; the results were compared between Schwann cells cultured in collagen sponge and culture dish. The scaffold carrying Schwann cells was implanted into the paraspinal muscle in rats, and H-E staining was used to observe the in vivo degradation and the inflammation responses. Results: The external diameters of the scaffold and micro-tubules were 3 mm and 100 μm, respectively, and the micro-tubules were arranged in an even and parallel manner. The adherence rates and proliferation rates of Schwann cells were similar between scaffold group and collagen sponge group, but both groups were significantly lower than the culture dish group (P<0.05). The 3D scaffold degraded completely within 12 weeks, with no visible inflammatory cells around. Conclusion: The self-designed 3D scaffold has no harmful effect on the growth of Schwann cells, and it can degrade in vivo, showing a satisfactory biocompatibility.

Researches on monoclonal gammopathy of undetermined significance (MGUS) and smoldering muhiple myeloma (SMM) are at a low level although their incidence is high.This review discusses curent biological insights in MGUS/SMM and discusses how the integration of novel biological markers,molecular imaging,and clinical monitoring of MGUS/SMM could facilitate the development of early treatment strategies for high-risk SMM (early myeloma) patients in the future.

Host-Pathogen Interactions ; Humans ; Immunocompromised Host ; MicroRNAs ; genetics ; immunology ; RNA, Viral ; genetics ; immunology ; Viruses ; genetics ; immunology

Acute Disease ; Adult ; Cardiomyopathies ; blood ; chemically induced ; diagnosis ; Creatine Kinase ; blood ; Female ; Humans ; Male ; Organophosphate Poisoning ; Pesticides ; poisoning ; Troponin I ; blood

Objective Insulin-like growth factor-1 receptor (IGF-1R),similar to insulin receptor,is one of the families of re- ceptor tyrosine kinases.,which has been found to be overexpressed in a variety of cancer.It is the main proliferation and survival sig- nal molecule in cancer cell and plays an important role in cancer growth and progress.Blocking signal transduction of IGF-1R by vari- ous strategies can suppress tumor growth and induce regression of established tumor.This study is to construct the siRNA expression vector targeting IGF-1R and to evaluate its ability to induce cell apoptosis in human lung cancer cells.Methods Two siRNA expres- sion vector,pENTR/U6-shRNA-1 and pENTR/U6-shRNA-2 targeting IGF-1R,were constructed using pENTR/U6 vector,and a vector targeting hieiferase gene,pENTR/U6-shRNA-Iuc,was constructed as control.After vectors were transfected into A549 for 48h, knockdown of IGF-1R mRNA and protein and Akt phosphorylation were accessed,and DNA ladder and flow cytometry were used for cell apoptosis.Results siRNA expression vectors targeting IGF-1R were successfully constructed,which was confirmed by PCR and DNA sequencing,pENTR/U6-shRNA-1 and pENTR/U6-shRNA-2 demonstrated the expression were (22.1?2.5) % and (80.1? 3.9) % in IGF-1R mRNA level,(15.2?3.1)% and (47.1?4.1)% in protein level,respectively,compared with pENTR/U6- shRNA-luc.Suppression of IGF-1R by pENTR/U6-shRNA-1 blunted Akt phosphorylation,increased cell apoptosis induced by 3% ethanol,and retained 77.5 % A549 cells in the G0/G1 phase.Conclusion siRNA expression vector targeting IGF-1R can effectively suppress the expression of IGF-1R expression in A549.This study suggests that DNA vector-based RNAi has the potential to be effec- tive and practical cancer gene therapy strategy.

Objective:To investigate whether transplantation of mesenchymal stem cells(MSCs)through renal arteries can protect kidney from acute tubular necrosis(ATN),so as to lay a foundation for MSC transplantation in treatment of ATN.Methods:Five-week-old nude mice were randomly divided into three groups:normal control group(n=10),acute tubular necrosis(ATN)model group without(n=10)and with MSCs treatment group(n=11).ATN nude mice were induced with 50% glycerin.MSCs labeled with enhanced green fluorescent proteins(EGFP)were injected into kidney through renal arteries.Serum creatinine was determined in all groups and pathological changes of renal tissues were detected using H-E staining.The amount and distribution of the EGFP-marked MSCs in renal tissues were determined with fluorescence microscope.Results:Degeneration and exfoliation of renal tubular epithelial cells,and even renal tubular tamponade with cast-off cells were observed in the ATN group;these pathological changes were mainly located at renal cortex and juncture of renal cortex and medulla.The damages were greatly alleviated in the ATN+MSCs transplantation group,with no swelling of epithelial cells,nuclear condensation or edema.Fourteen days after MSCs transplantation,EGFP positive cells were increased in renal tubules of recipient mice.Conclusion:The MSCs transplantation via renal artery can locate in renal tubular epithelium,and promote the repair of injured renal tubular epithelial cells.

There is a variety of gut microbiota in human body, which is closely associated with the health and disease. Normal gut microbiota can produce colonization resistance to pathogens. Antibiotics can affect the composition of gut microbiota and change the intestinal microenvironment, resulting in intestinal microecological disorders, which in turn cause intestinal pathogenic infections and other diseases. In this paper, the concept of intestinal microecology, the mechanism of intestinal colonization resistance, the effect of antibiotics on intestinal microecology, and the treatment methods were reviewed, aiming to provide the information for the rational use of antibiotics and the development of more effective treatment methods to maintain the stability of intestinal microecology.

Objective The aim of the study WSfl to establish the method using CDl27 as the new biomarker to identify regulatory T cells(Treg cells)and apply the CD 127 to detect the Treg cells in patients with gastric cancer.Methods The phenotypes of Treg cells were analyzed using five-color flow cytometry method Foxp3.FITC/CD127-PE/CD4-PerCP/CD25-APC/CD3-PC7.The mRNA and protein expression of Foxp3 in isolated CD+4 CD25high CD127-/low Treg cells were detected.The relationships between Foxp3 and CD127 protein expression in CD4+ T cells from aduh human peripheral blood were investigated.PBMCs,Ascltes,turnor-infihration lymphocyte and tumor-draining lymph nodes in 35 patients with gastric cancer and PBMCs in 20 normal healthy donors were evaluated for the proportion of Treg ceils,as well as the percentage ot the total CD +4 cells.Results CD4+ CD25 high CD-127 low ceils expressed the high Foxp3 in protein level(87.1%)and mRNA level.Within the CD4+CD+25 population,there was a significant correlation between Foxp3 and the CD127low phenotype(r=0.985,P<0.01).Compared with healthy olunteers,patients with gastric malignancies had a higher proportion of CD4+4 Cdhigh 25 CD-low127 cells in peripheral blood(t=2.542,P<0.05).The Dereentages of Treg cells were more abundant in ascites(t=2.357,P<0.05),TIL(t=6.174,P<0.01) and tumor-draining lymph nodes(t=5.481,P<0.01)of individuals with gastric cancer than that in their blood.There were significant differences in the prevalence of Treg ceils between the early and advanced disease stages in gastric cancer[(6.04±2.31)%in stageⅠ+Ⅱ VB(10.16±2.29)% Ⅲ+Ⅳ,t=2.473,P<0.05].Conclusions The CD127 biomarker can be used to selectively enrich human Treg cells for in vitro functional studies.The populations of CD4+ CD high25 CD127-/low Treg cells increased ith tumor stage in individuals with gastric cancer.

Objective To investigate the populations of CD+4CD25high Foxp3+ regulatory T cells(Treg cells)and mRNA expression of Foxp3 in peripheral and the marginal region of tumor from patients with gastric cancer,and to evaluate the prevalence of Treg cells from patients with gastric cancer in relation to the TNM stages.Methods PBMC,Ascites,tumor-infiltration lymphocyte and tumor-draining lymph nodes in 47 patients with gastric cancer(TNM I11,Ⅱ18,Ⅲ10,Ⅳ8)and PBMC in 20 normal healthy donors were evaluated for the proportion of Treg cells,as a percentage of the total CD4+ cells,by flow cytometric analysis.Levels of mRNA for Foxp3 were measured witll a real-time quantitative PCR Results The percentage of CD4+CD25high Foxp3+ Treg cells in PBMC for cases of gastric cancer were significantly higher than those for healthv donors(P<0.05).The percentage of Treg cells were more abundant in ascites(P< O.05),1rIL(P<0.01)and tumor-draining lymph nodes(P<0.01)of individuals with gastric cancer than in their blood.And the Foxp3 mean fluorescent intensity (MFI) of Treg cells in TIL and tumor draining lymph nodes with gastric caners were significantly higher than PBMC and ascites(P<0.05).The proportion 0f Treg cell in patients of gastric cancers with stage I,II,Ⅲ,Ⅳ were(5.72±1.95)%,(6.45± 2.23)%,(9.58±3.13)%,(11.70±2.30)%,respectively.There were significant correlation between Drevalenee of Treg cells and disease stages in gastric cancer(r=0.784,P<0.01).Foxp3 mRNA levels were higher in PBMC from the group with gastric cancers than in the group with healthy donors.Foxp3 mRNA levels were correlated with TNM stages(r=0.623,P<0.05).Conclusions Our results suggest that the populations of CD4+CDhigh25 Foxp3+Treg cells and Foxp3 mRNA levels in patients with gastric cancer are signifieantlv higher in comparison to those in control donors.In addition,the expression of Fox：p3 mRNA increased with tumor stage.The increased prevalence of Treg cells may be one of the explanations for impaired cell-mediated immunity in cancer bearing hosts.