0.05). CONCLUSION: Both modified Dexter method and IMDM medium method can promote the adherence and proliferation of bone marrow stromal cells in a short time. It is the simple and practical culture method of bone marrow stromal cells by culturing the cells in the medium of IMDM compared to the modified Dexter method.

Objective:To study of isolating culture and differentiation of human bone marrow-derived mesenchymal stem eeUs into blood vessel endothelial-like cells in a specialized micro-environment in vitro,SO as to provide an experimental foundation for psoriasis.Methods:The hMSCs were isolated by density gradient centrifugation,amplificated and identificated in vitro.Vascular endothelial growth factor (VEGF)and basic fibroblast growth factor(bFGF)within endothelial cell growth medium(DMEM)were used to induce hMSCs differentiation into vascular endothelial-like cells.The induced hMSCs were detected by flow cytometry to find whether they had endothelial cell phenotypes.The Dil-ac-LDL ingestion assay Was used to apprmses the blood vessel endothelial-like cell function.Results:In cell morphology,the induced hMSCs transformed into endothelial-like cells.These cells expressed specific surface markers of,Vascular endothelial-like cells such as CD34,CDl06,HLA-DR,CD54,VWF,CD31,KDR and CD5 comparing to those in the control group(P＜0.01).The induced endothelial-like eeHs had the ability of ingesting Dil-ac-LDL.Conclusion:Combination of Density gradient eentrifugation and adherent methods can obtain pure MSCs.hMSCs Can obtain endothelial cell phenotypes after induced by VEGF and bFGF in vitro.Human hnSCs have potential to differemiate into vascular endothelial-like cells.The induced endothelial-like cells have completely mature endothelial cell functional properties.

BACKGROUND:It is discovered in our preliminary study that the activity of psoriatic bone marrow hematopoietic cells is abnormal.OBJECTIVE:To reveal level of stem cell factor(SCF) and granulocyte colony-stimulating factor(G-CSF) secreted by bone marrow stromal cells in psoriasis patients.DESIGN,TIME AND SETTING:This case-control observation experiment was performed at the Laboratory of Department of Dermatology,Taiyuan Central Hospital from October 2007 to August 2008.PARTICIPANTS:Twenty-four common-psoriasis outpatients were selected at the Department of Dermatology,Taiyuan Central Hospital,including 16 males and 8 females,aged 16-59 years old.Twenty controls were selected from Department of Hematology,Taiyuan Central Hospital,whose age and gender were matched with psoriasis patients.METHODS:Bone marrow mononuclear cells were isolated from psoriasis patients and controls using the density gradient centrifugation.Bone marrow stromal cells were cultured by the adherent method.When the cells were cultured for three passages and 72 hours,bone marrow stromal cells and supernatant liquid were collected.MAIN OUTCOME MEASURES:The phenotypes of cells were identified by flow cytometry and the level of SCF and G-CSF were detected by enzyme linked immunosorbent assay kit(ELISA).RESULTS:Flow cytometry assay showed that over 90% bone marrow stromal cells were positive for CD29,but negative for CD34,CD45 and HLA-DR.That is,the purity of bone marrow stromal cells was over 90%.The levels of SCF and G-CSF in psoriasis patients were significantly higher than that in controls(P

Objective To investigate the effect of brain microvascular endothelial cells (BMECs) on mesenchymal stem cells (MSCs) expressing fetal liver kinase-1(Flk-1) and von Willebrand factor (vWF) under normal and hypoxic conditions. Methods Human bone marrow MSCs and rat BMECs were isolated, cultured and identified. We used direct and indirect co-culture to induce the differentiation of MSCs. The expression of Flk-1 and vWF after induction was observed and analyzed by flow cytometry and immunofluorescence staining. Results Undifferentiated MSCs did not express Flk-1 or vWF. In indirect co-culture group, indirect co-culture under hypoxic conditions induced (7.58?0.58)% (n=6, P=0.034) MSCs to express Flk-1 but not vWF, while in normoxic indirect co-culture, MSCs expressed neither. In direct co-culture group, after 5-day direct co-culture with BMECs under normoxia or hypoxia, MSCs expressing Flk-1 accounted for (13.76?1.67)% (n=6, P

Objective To study the risk factors of complications of 364 cases in treatment of rad-ical esophagectomy.Methods 364 patients underwent two or three-incision radical esophagectomy and the clinic data were retrospectively analyzed.After review of the reference,ten factors were regarded as the potential risk factors of postoperative complications,then univariate and multivariate analyses were per-formed.Results In 364 patients,66 patients had complications of this research field within postoperative 6 months.By univariate analysis,preoperative nutritional risk score、T-staging、preoperative complications、operation time、operation experience、anastomotic position、anastomotic extra manual suturing were deter-mined as influence factors.By multivariate analysis,preoperative nutritional risk score≥3、T-staging score≥3、preoperative complications、operation time≥240 min、cervical anastomosis were determined as risk factors,anastomotic extra manual suturing was determined as a protective factor.Conclusion The risk factors of the complications after radical esophagectomy are preoperative nutritional risk score≥3、T-stag-ing score≥3、preoperative complications、operation time≥240 min、cervical anastomosis;and the protective factor is anastomotic extra manual suturing.Paying attention and controling these risk factors may reduce the occurrence of postoperative complications.

AIM:To investigate the paracrine actions of mesenchymal stem cells(MSCs)to brain microvascular endothelial cells(BMECs)in hypoxic conditions.METHODS:Human bone marrow MSCs and rat BMECs were isolated,cultured and identified.The contents of VEGF and MMP-9 were confirmed using ELISA assays of the conditioned media.The transendothelial electrical resistance(TEER)was detected to reflect the permeability of the BMECs monolayer.The effects of different conditioned media on BMECs were investigated in hypoxic conditions.RESULTS:The contents of VEGF and MMP-9 in conditioned media were induced to increase significantly by the hypoxic stress,which were much lower in BMECs conditioned media than that in MSCs conditioned media.Hypoxia induced a significant increase in VEGF and MMP-9 in MSCs conditioned media collected in comparison to those in normal conditions.In hypoxic conditions,MSCs conditioned media enhanced the proliferation(optical density values:0.947?0.103 vs 0.254?0.024,P

Objective To evaluate the inhibitory effect of mesenchymal stem cells (MSCs) in lesions of patients with psoriasis on T lymphocyte proliferation.Methods Tissue specimens were obtained from the lesions of 15 patients with psoriasis vulgaris (7 at progressive stage and 8 at resting stage) and normal skin of 15 human controls from the Department of Urology and Plastic Surgery,Taiyuan City Centre Hospital.MSCs were isolated from these skin specimens,cultured,and identified using flow cytometry and in vitro differentiation assay.Enzyme-linked immunosorbent assay (ELISA) was performed to detect the concentration of interleukin (IL)-6,IL-1 1,hepatocyte growth factor (HGF) and transforming growth factor (TGF)-β1 in the culture supernatant of third-passage MSCs.Peripheral blood T cells were obtained from a healthy adult and cocultured with the third-passage MSCs for four days.Then,cells were counted and methyl thiazolyl tetrazolium (MTT) assay was conducted to evaluate the proliferation of T cells.One-way analysis of variance (ANOVA) and Student-Newman-Keuls (SNK) test were carried out to compare the proliferation of T lymphocytes,and two independent samples t test to compare the concentrations of cytokines.Results Inverted microscopy revealed that the patient-and control-derived MSCs shared similar morphological properties and multi-directional differentiation capacity,along with the expression of CD29,CD44,CD73,CD90 and CD105,but absence of CD34,CD45 and HLA-DR on cell surface.After coculture with MSCs from the patients and controls for four days,the count of T lymphocytes per milliliter was (1.67 ± 0.34) × 105 and (1.04 ± 0.29) × 105 respectively (P＜ 0.01),and the proliferative activity (expressed as absorbence at 492 nm)was 0.317 ± 0.021 and 0.275 ± 0.007 respectively (P ＜ 0.01).Compared with the control-derived MSCs,the patient-derived MSCs showed a significantly higher level of IL-1 1 ((181.37 ± 31.74) vs.(130.07 ± 29.20) ng/L,t =5.32,P ＜ 0.01),but a lower level of lL-6 ((61.67±17.53) vs.(76.74±18.96) ng/L,t=2.61,P＜0.05)and HGF ((319.24 ± 41.03) vs.(352.35 ± 51.47) ng/L,t =2.25,P＜ 0.05),as well as a similar level of TFG-β1,in the culture supernatant.Conclusions The inhibitory effect of MSCs in psoriatic lesions on T lymphocyte proliferation is diminished,which may contribute to the pathogenesis of psoriasis.

Objective To discuss whether Saccharomyces boulardii sachets can improve the eradication rate and reduce some untoward effects of gastrointestinal tract when adding in the Helicobacter pylori (Hp) standard quadruple therapy.Methods 249 cases infected with Hp were randomly divided into control group(124 cases) and treatment group (125 cases).The control group was treated with standard quadruple therapy:colloidal bismuth pectin capsules,100 mg,tid,po;furazolidone tablets,0.1 g,bid,po;amoxil capsule,1 g,bid,po;rabeprazole sodium enteric-coated capsules,20 mg,qd,po.The treatment group was treated with Saccharomyces boulardii sachets(0.25 g,bid,po)besides.Both groups were treated for 4 weeks.Evaluate the effect of Hp eradication rate by using the method of intent of treatment (ITT) analysis and consistent scenario set (PP)analysis.Results According to the ITT analysis,the eradication rate of control group was 89.52%,and that of treatment group was 96.00%.According to the PP analysis,the eradication rate of control group was 90.00%,and the rate of treatment group was 97.39%.Both analysis results showed the eradication rate of Hp in treatment group was significantly higher than control group(P<0.05).As for reducing the untoward effects such as nausea,inappetence,diarrhea,astriction and erythra,control group was obviously lower than treatment group (P<0.05).Conclusion Compared with standard quadruple therapy only,adding Saccharomyces boulardii sachets can significantly improve the eradication rate of Hp and obviously reduce the untoward effects such as nausea,inappetence,diarrhea,astriction and erythra.It is worth to be generalized and applied in clinical practice.

Objective To investigate and analyze of the distribution of herpes zoster in in-patients from 1995 to 2004.Methods The datas of in-patients with herpes zoster from 1995 to 2004 were statistically analyzed.Results There were 290 primary diseases,154 complications and 335 without underlying diseases.In-patients with herpes zoster were increasing year by year.The amount of severe patients over 60 years old were more than these of other old group.Conclusions The incidence of herpes zoster are rising year by year.There is high incidence and complications and more postherpetic neuralgia in old man and the people with immune disorder.So early diagnosis and treatment is an effective preventing and treating measure.

Objective To study the in vitro activity of T cells differentiated from bone marrow CD34+ cells of psoriatic patients with family histories. Methods Bone marrow CD34+ cells were isolated from psoriatic patients with family history and normal persons by immunomagnetic cell selection and induced to differentiate to T cells in thymic stromal microenvironment in vitro. CD3+ T cells were obtained by CD3+ cell selection system and the proportions of CD4+CD8- and CD4-CD8+ cells were measured by flow cytometry. The proliferation activity of T cells was measured by MTT colorimetry in both spontaneous proliferating group and superantigen (SAg)-stimulated group. And the levels of IL-4, IL-8 and IFN-? were quantified by enzyme-linked immunosorbent assay in culture supernatant. Results Mature CD4+CD8- cells and CD4-CD8+ cells were detected from CD3+ cells, which were derived from bone marrow CD34+ cells. When the psoriatic patients and normal controls were compared, there was no significant difference in the proportion of CD4+CD8- cells or that of CD4-CD8+ cells in CD3+ cells. The proliferation activity of T cells was significantly higher in the psoriatic patients than that in the normal controls. There was no statistical difference of IL-4?IL-8 or IFN-? levels in the supernatant of T cells between the spontaneous group and the normal control. However, the IFN-? and IL-8 levels in the supernatant of T cells were higher in the psoriatic patients than those in the normal controls after the SAg stimulation. Conclusion The abnormal activity of peripheral blood T cells from psoriatic patients with family history may be related to the bone marrow hematopoietic cells.