Objective To find a safe and effective way for the collection of stem cells from peripheral blood in low body weight children. Methods Peripheral blood stem cells were collected altogether 15 times in 5 children with body weight lower than 20 kg. One day before the collection, a double cavity tube specially made for children was inserted into femoral vein and remained there for future use. Peripheral blood stem cells were separated by the separation vessel specially designed for children and collected in the collecting tank for children. After initialization, alien blood irradiated by X ray was infused in and moved through the separation machine to keep the blood balanced and hematocrit stable in the body of children. During the separation, anticoagulant/whole blood was kept between 1∶11 and 1∶13. The volume of blood processed every time was 2-4 times of total volume of child blood(2 700-5 850 ml). Results Mononuclear cells and CD34 + cells were successfully collected each time in all 5 children, enough for the transplantation of peripheral blood stem cells. Conclusion The way we developed to collect peripheral blood stem cells in children with low body weight is safe and effective.

Objective: To assess the immunological effects by orally administering chicken type Ⅱ collagen(CCⅡ) on collagen-induced arthritis(CIA)mice. To assess the effect on producing IL-1 of peritoneal macrophage in adjuvant arthritis rats by orally administering CCⅡ. Methods: Arthritis were induced in Kunming mice by immunization with chicken type Ⅱ collagen with Freund's complete adjuvant, followed by an interperitoneal injection of CCⅡ 3 weeks later.Chicken type Ⅱ collagen was orally administered from 5 days prior to the induction of arthritis to 14 days after inducing arthritis model. The animals were examined visually twice weekly for polyarthritic signs of swollen and erythemic limbs. Quantitation of antibody level of CIA mice was measured by ELISA method. Subpopulations of T lymphocytes in mice were evaluated by flow cytometry method. IL-1 assay was evaluated by ELISA method. Results: Joint swelling was significantly reduced at a dose of 5 μg.kg-1 and 50 μg.kg-1 of CCⅡ, but not at 250 μg.kg-1. The level of anti-collagen antibodies was also reduced at a dose of 5 μg.kg-1 and 50μg.kg-1 (OD value from CIA model control 0.242±0.073 to CCⅡ 5 μg.kg-1 0.123±0.029 and CCⅡ 50 μg.kg-1 0.110±0.075 respectively). Subpopulations of T-lymphocytes were changed by orally administering of CCⅡ, and the ratio of L3T4+/Lyt-2+ was lowered (the ratio from 1.71 of CIA model control to 1.21, 1.51 of administered CⅡ 5 μg.kg-1, 50μg.kg-1 respectively.) after administering CCⅡ. IL-1 level can be reduced (the value from adjuvant arthritis model control 62.8±0.9 to 43.4±1.3, 49.7±0 ng.L-1 administered CⅡ 5 μg.kg-1, 50μg.kg-1 respectively). Conclusion: Arthritis sign in CIA animal model can be suppressed by oral CCⅡ. The effects may be involved by influencing the mechanisms both humoral and cellular immunity. The effects occurred at lower doses of CCⅡ. These results demonstrated the biologic relevance of by-stander suppression associated with oral tolerance, and the potential use of this approach to treat human inflammatory joint diseases.

AIM: To discuss the influence of high glucose on the proliferation and ICAM-1 expression of human umbilical vein endothelial cells(HUVEC) within 48 h and to observe the altered effect of high glucose when high glucose and inflammatory stimulator exist at the same time.METHODS: HUVEC were cultured with different concentrations of glucose.The proliferation of HUVEC induced by high glucose was detected by MTT assay.The ICAM-1expression of HUVEC induced by high glucose was assayed by flow cytometry and cell ELISA.The difference of ICAM-1 expression between induced by LPS and by LPS together with high glucose was compared.RESULTS: 25、(45 mmol?L~(-1)) glucose did not inhibit the proliferation of HUVEC within 48 h,and they also had no effect on the expression of ICAM-1.However,when stimulated HUVEC with LPS, the proliferation of HUVEC was extremely inhibited in high glucose groups,and the expression of ICAM-1 increased significantly in those groups.CONCLUSION: In short time,high glucose has no effect on the proliferation and ICAM-1 expression of HUVEC,but when inflammatory stimulation exists,high glucose can inhibit the proliferation and increase the expression of adhesion molecules.

Aim To observe the effect of Sinomenine on the activation of NF-?B and the degradation of its inhibitor I-?B in peritoneal macrophages of BALB/c mice in vitro and provide experimental evidence for further evaluation of the antiinflammatory and antirheumatic effects of Sinomenine. Methods Effect of Sinomenine on the activation of NF-?B p65 in the cells was investigated by using fluorescence-labelling and laser confocal scanning microscopy; Effect of Sinomenine on the degradation of I?B-? was investigated by Western blot. Results SIN (0.25,1.25 mmol?L -1) attenuated the activation of NF-?B p65 in peritoneal macrophages of BALB/c mice induced by LPS in vitro. SIN(0.25,1.25 mmol?L -1) inhibited the degradation of I-?B-? in peritoneal macrophages of BALB/c mice induced by LPS in vitro. Conclusion SIN can partially inhibit the activation of NF-?B p65 and the degradation of I?B-? in peritoneal macrophages of BALB/c mice induced by LPS in vitro.

Objective To compare the transfection efficiency of cationic polyethylenimine(PEI)with Lipofectamine 2000TM by using the plasmid DNA encoding vascular endothelial cell growth factor (VEGF165 )gene in human embryonic kidney cell line 293T.Methods PEI of different N/P ratio and Lipofectamine 2000TM were used to deliver the vector containing VEGF165 to 293T cells,respectively.Green fluorescent protein(GFP)gene was inserted into the vector as a report gene.Evaluation of cytoactive was performed by CCK-8 assay 24 h after transfection.The cells were observed by fluorescent microscope and the presence of VEGF165 in cell supernatant was detected by ELISA 48 h after transfection.The transfection efficiency was calculated and com-pared.Results Similar cytoactive and best transfection efficiency could be obtained when N/P ratio was 9,the transfection efficien-cy was around 70%.Furthermore,the presence of VEGF165 increased significantly after transfection(P <0.05),but there was no significant difference between the two groups in which different transfection methods were adopted.Conclusion PEI as a novel oli-gofectamine reagent could mediate more efficient transfection compared with lipofectamine.It also has low cell-toxicity and low price and could be an ideal vector in gene delivery technology.

Objective To investigate the association of serum leptin (LEP) levels during the first postpartum year with the occurrence of postpartum thyroiditis (PPT).Methods Fifty-seven PPT patients consisted of 34 with overt PPT and 23 subclinical PPT.37 healthy postpartum women were used as controls.Serum samples were obtained at 4 postpartum date points,i.e.3-days and 3,6,12-months postpartum.LEP level was determined by radioimmunoassav.Results Compared with control women,PPT patients were maintaining significantly higher levels of LEP and LEP/body mass index (BMI) ratio during the first postpartum year.There was no significant difference in serum LEP level or LEP/BMI ratio between overt PPT and subclinical PPT groups.In PPT patients,LEP and LEP/BMI ratio were negatively correlated with serum TSH,and positively correlated with serum FT4 and FT3.Conclusion Sustained high levels of serum LEP after delivery may favor the occurrence of PPT.Further studies are needed to clarify the specific role played by LEP in PPT.

Objective To investigate the feasibility of green fluorescent protein (GFP) as a marker to trace the transplanted um-bilical cord mesenchymal stem cells (ucMSCs) in rats with cerebral ischemia-reperfusion .Methods ucMSCs were transfected by GFP-adenovirus .The rats were subjected to left middle cerebral artery occlusion using suture method .1 × 106 GFP-ucMSCs were transplanted with cerebral stereotaxic technique .Frozen sections of brain tissue were made at 7 d after cerebral ischemia .The ex-pression of GFP was observed by fluorescence microscope .Results In vitro ,the morphology of GFP in ucMSCs was fibrous ,and when fused ,the clusters were arranged in a radial or whirlpool shape ,The morphological feature of transfected ucMSCs was similar to that un-transfected ucMSCs .Under the fluorescence microscope ,the positive rate of GFP was more than 80% .In addition ,GFP could spread to the whole cells and show the completed form .No rejection was observed in the rats after transplantation ,and the GFP was found near the site of transplantation after 7 d ,and the fluorescence was not attenuated .Conclusion GFP is an effective tracer maker for ucMSCs transplantation in the treatment of ischemia-reperfusion ,which could provide experimental method for fur-ther study .

Pulmonary hypertension (PH) is an insidious pulmonary vasculopathy with high mortality and morbidity and its underlying pathogenesis is still poorly delineated. The hyperproliferation and apoptosis resistance of pulmonary artery smooth muscle cells (PASMCs) contributes to pulmonary vascular remodeling in pulmonary hypertension, which is closely linked to the downregulation of fork-head box transcriptional factor O1 (FoxO1) and apoptotic protein caspase 3 (Cas-3). Here, PA-targeted co-delivery of a FoxO1 stimulus (paclitaxel, PTX) and Cas-3 was exploited to alleviate monocrotaline-induced pulmonary hypertension. The co-delivery system is prepared by loading the active protein on paclitaxel-crystal nanoparticles, followed by a glucuronic acid coating to target the glucose transporter-1 on the PASMCs. The co-loaded system (170 nm) circulates in the blood over time, accumulates in the lung, effectively targets the PAs, and profoundly regresses the remodeling of pulmonary arteries and improves hemodynamics, leading to a decrease in pulmonary arterial pressure and Fulton's index. Our mechanistic studies suggest that the targeted co-delivery system alleviates experimental pulmonary hypertension primarily via the regression of PASMC proliferation by inhibiting cell cycle progression and promoting apoptosis. Taken together, this targeted co-delivery approach offers a promising avenue to target PAs and cure the intractable vasculopathy in pulmonary hypertension.