Objective To observe the effects of metformin on expression of Adenosine 5'-monophosphate (AMP)-activated protein kinase (AMPK),nuclear factor-κB (NF-κB) and transforming growth factor β1 (TGF-β1) in cultured rat glomerular mesangial cells (MCs),and explore its renoprotective mechanisms.Methods MCs were cultured in the medium with normal glucose (group NG,5.6 mmol/L),high glucose (group HG,25mmol/L) and different concentrations of metformin (group M1,M2,M3).After 48 h exposure,the supernatants and MCs were collected.The expression of NF-κB and TGF-β1 mRNA was analyzed by real time-PCR.Total-AMPK,phospho-Thr-172 AMPK (p-AMPK),NF -κB p65 and TGF-β1 were visualized by Western blot.Results The real time-PCR and Western blot result showed MCs could express AMPK,NF-κB and TGF-β1 mRNA and protein.After stimulated by HG,the levels of intracellular NF-κB and TGF-β1 expressions were significantly increased compared with group NG (P ＜ 0.05); The levels of NF-κB and TGF-β1 were significantly decreased in group M1,M2 and group M3 compared with group HG in a dose-dependent manner.After stimulated by HG,the level of intracellular p-AMPK were down-regulated compared with group NG(all P ＜ 0.05); The expression of p-AMPK increased with the rising of metformin concentration,presenting the opposite trend (P ＜ 0.05),while the level of total-AMPK protein was unchanged with exposure to HG or different concentrations of mefformin(P ＞ 0.05).Conclusion Metformin can suppress the expression of NF-κB and TGF-β1 of glomerular MCs induced by HG via AMPK activation,which may partly contribute to its reno-protection.

Objective:To explore the role of static posturography in the assessment of fatigue due to flight tasks.Methods:Thirtymale college students were asked to perform simulated flight tasks consecutively forfour hours.Meanwhile their statie posturography and tasks performance would be repeatedly measured during the task-load at end of every hour.Based on the changed significantly parameters,the static balance index would be built by principle component analysis.Then its correlation with task-load level would be further analyzed by curve estimation.Results:Static postural control declined significantly under effect of simulated flight tasks.With task load sustaining,static balance index increased significantly and correlated linearly with duration of task load (R2=0.949).Besides,there was quadratic relationship between the change of multi-tasks performance and duration of task load (R2=0.968).And correlation of multi-tasks performance with static standing balance level also had been proved to be quadratic (R2=0.976).Conclusions:Static posturography correlated linearly with flight task-load level,which could reflect fatigue level caused by task load.

OBJECTIVETo explore the improvement of dendritic cells' (DCs) functions in chronic hepatitis B (CHB) patients by two different drugs plasma, i.e., Shen supplementing and detoxification (SSD) and Pi invigorating and detoxification (PID), thus comparing which method was more effective to activate DCs to improve T lymphocyte functions.

METHODSTotally 30 CHB outpatients were recruited. They were assigned to the immune tolerant group and the immune clearance group, 15 in each group. Totally 60 mL peripheral blood was extracted to isolate and develop mature DCs. Chinese compound containing (Liuwei Ganlu Syrup for SSD) and (Sijun Ganlu Syrup for PID) plasma were added to promote the maturation of DCs on the 7th day. Besides, non-drug plasma was taken as the control. On the ninth day, HBV core 18-27 loaded core peptide and its own T lymphocyte were co-cultivated for 72 h. Then T lymphocytes were collected. The expression levels of CD3, CD28, CD4, and CD8, programmed death-1 (PD-1) were detected using flow cytometry.

RESULTSCompared with non-drug plasma, the expression levels of CD3, CD4, and CD28 could be improved, and the expression levels of CD8 and PD-1 could be reduced by the two methods, showing statistical difference (P < 0.05). Besides, SSD containing plasma showed better effect in improving the molecular CD28 expression rate, and reducing the molecular PD-1 expression rate on the T cell surface, showing statistical difference when compared with that of PID containing plasma (P < 0.05).

CONCLUSIONSIn vitro intervention of DCs by SSD and PID containing plasmas combined co-cultivation of its own T lymphocytes could promote the activation of DCs to improve the function of T cells and the expression of T cell surface molecules. Besides, SSD showed more significant effect on infection immune of HBV patients in the tolerance stage.

Adult ; Cells, Cultured ; Dendritic Cells ; drug effects ; immunology ; Drugs, Chinese Herbal ; pharmacology ; Female ; Hepatitis B, Chronic ; immunology ; Humans ; Male ; Middle Aged ; T-Lymphocytes ; immunology ; Young Adult

BACKGROUND: Immunoloregulation of mesenchymal stem cells (MSCs) is commonly approved. Previous studies have confirmed the ability of Flk-1~+ bone marrow MSCs (BMSCs) to inhibit T/B lymphocyte proliferation in vitro. OBJECTIVE: To investigate the therapeutic effect of Flk-1~+ BMSCs in collagen-induced arthritis mice.METHODS: A total of 18 healthy male DBA-1(H-2K~q) mice aged 10 weeks were randomly divided into 3 groups. All the mice were injected at the base of the tail with bovine type II collagen (CII), and received a booster injection of CII on day 21 to establish the CIA mice model. DBA-1(H-2K~q)mouse Flk-1~+ BMSCs were isolated in vitro by the density gradient centrifugation and adherence screening. Following initial immunity, mice in the cell transplantation group were infused with Flk-1~+ BMSCs (1-2)×106 cells/mouse via the caudal vein. Mice in the cell transplantation group were injected with the same volume of Flk-1~+ BMSCs during booster. Mice in the model control group were injected with an equal volume of saline 0 or 21 days following initial immunity. Following initial immunity and booster immunization, claw pad thickening and clinical score were observed, changes of joint pathology and dynamic changes in serum factor mass concentration were determined in mice. RESULTS AND CONCLUSION: Compared with the model control group, no significant difference in claw pad thickening and mean clinical score was detected in the cell transplantation group following initial immunity (P > 0.05), with the presence of obvious damage to synovial membrane and inflammatory cell infiltration. Mass concentration of each serum cell factor was similar. The claw pad was significantly thickened (P < 0.01), mean clinical score reached 3.35 points, with severe damage to synovial membrane, proliferation of blood capillary in the cell transplantation group following booster immunization. Interleukin-6 levels were greatly increased at day 28 following initial immunity (P < 0.1), but decreased at day 35 following initial immunity (P < 0.1). Results indicated that in the collagen-induced arthritis mouse models, Flk-1~+ BMSC transplantation did not obtain prospective therapeutic efficacy, but aggravation of arthritis was observed in the cell transplantation group following booster immunization. Upregulation of interleukin-6 concentration could aggravate the behavior symptom of rheumatoid arthritis mice.

Objective To observe the effects of intensive lower extremity motor control training on the motor function of lower extremity, balance and walking capability of stroke patients. Methods 40 stroke patients were randomized into the intervention group (n=20) and control group (n=20). Both groups received the routine rehabilitation, while the intervention group accepted the lower extremity motor control training in addition for 6 weeks. Before and after 6 weeks of treatments, they were assessed with Fugl-Meyer Assessment of lower extremity (FMA-L), Berg Balance Scale (BBS), Holden Functional Ambulation Category (FAC); their gaits were analysesd with footprint. Results Both groups improved significantly in scores of FMA-L, BBS, FAC and average step length, stride width, walking velocity (P<0.01) after treatment, and improved more in the intervention group than in the control group (P<0.05). Conclusion Intensive motor control training of lower extremity may promote the recovery of motor function of lower extremities, balance and walking ability of stroke patients.

BACKGROUNDOxidative Stress and p38 mitogen-activated protein kinase (p38MAPK) play a vital role in renal fibrosis. Pioglitazone can protect kidney but the underlying mechanisms are less clear. The purpose of this study was to investigate the effect of pioglitazone on oxidative stress and whether the severity of oxidative stress was associated with the phosphorylation level of p38MAPK.

METHODSRat mesangial cells were cultured and randomly assigned to control group, high glucose group and pioglitazone group. After 48-hour exposure, the supernatants and cells were collected. The protein levels of p22(phox), p47(phox), phosphorylated p38MAPK, total p38MAPK were measured by Western blotting. The gene expressions of p22(phox), p47(phox) were detected by RT-PCR. The levels of intracellular reactive oxygen species (ROS) were determined by flow cytometry. The levels of superoxide dismutase (SOD) and maleic dialdehyde (MDA) in the supernatant were determined respectively.

RESULTSCompared with the control group, the expression levels of p22(phox), p47(phox), phospho-p38 and ROS significantly increased, activity of SOD decreased in high glucose group, while the level of MDA greatly increased (P < 0.01). Pioglitazone significantly suppressed p22(phox), p47(phox) expressions and oxidative stress induced by high glucose. The expressions of p22(phox), p47(phox), phospho-p38MAPK and ROS generation were markedly reduced after pioglitazone treatment (P < 0.05). The activity of SOD in the the supernatant increased (P < 0.05), while the level of MDA decreased greatly by pioglitazone (P < 0.05). The level of oxidative stress was associated with the phosphorylation level of p38MAPK (P < 0.01).

CONCLUSIONPioglitazone can inhibit oxidative stress through suppressing NADPH oxidase expression and p38MAPK phosphorylation.

Animals ; Blotting, Western ; Glucose ; pharmacology ; Mesangial Cells ; drug effects ; enzymology ; NADPH Oxidases ; genetics ; metabolism ; Rats ; Reactive Oxygen Species ; metabolism ; Thiazolidinediones ; pharmacology ; p38 Mitogen-Activated Protein Kinases ; genetics ; metabolism

Objective To evaluate the immune effects of bivalent inactivated rotavirus vaccine (IRV) and investigate the viability of development of bivalent IRV.Methods Firstly,bivalent IRV was prepared by mixing G1 IRV and G3 IRV with equal amount,G1 IRV and G3 IRV as monovalent control,PBS as negative control.Secondly,those vaccines were vaccinated to the mice by intramuscular injection.Then,to evaluate the immune effects of bivalent IRV,the levels of serum or fecal rotavirus specific IgG and IgA were assessed by ELISA,the levels of serum neutralized antibody were measured by microneutralization assay,the number of IFN-γ or IL-4 secreting cells were analyzed by ELISPOT assay.Results Compared to negative control group,bivalent IRV induced the higher levels of serum and fecal G1 and G3 rotavirus specific antibody.It was found that there were no significant differences for the levels of serum IgG and IgA,fecal IgG and IgA,serum neutralized antibody between induced by bivalent IRV and induced by G1 type monovalent vaccines ; but there were significantly increase for the levels of serum IgG (t =2.691,P＜0.05) and serum neutralized antibody (t =2.561,P＜0.05) between induced by bivalent IRV and induced by G3 monovalent vaccines,there were no significant differences for other antibodies between induced by bivalent IRV and induced by G3 monovalent vaccines.At the same time,compared to negative control group,bivalent IRV induced significantly increase in the number of IFN-γ or IL-4 secreting cells in spleen lymphocytes.It was found that there were no significant differences for the number of IFN-γ or IL-4 secreting cells stimulated by G1 rotavirus between bivalent IRV and G1 monovalent vaccines; but there were significantly increase for the number of IL-4 secreting cells (t =2.327,P＜0.05) stimulated by G3 rotavirus between bivalent IRV and G3 monovalent vaccines,there were no significant differences for the number of IFN-γ secreting cells stimulated by G3 rotavirus between bivalent IRV and G3 type monovalent vaccines.Conclusion The bivalent IRV can induce effective immune response,in which there were no inhibitory interference between the components of bivalent IRV,which provided the experimental basis for the development of bivalent IRV.

ObjectiveTo observe the influence of different dietary protein intake (DPI) on nitrogen balance and nutritional indices in peritoneal dialysis (PD) patients, and explore the minimal DPI to maintain nitrogen balance.MethodsThirty-four PD patients were randomly divided into group A, B and C with DPI as 1.2, 0.9 and 0.6 g·kg-1·d-1 respectively. All the patients admitted into our hospital and completed a 10-day assessment for nitrogen balance, as well as nutritional status including serum albumin (Alb), pre-albumin at baseline, the 7th and 10th day. ResultsThe DPI of group A, B and C was (1.18±0.05), (0.87±0.02), (0.66±0.03) g·kg-1·d-1, whose differences were significant (P<0.01). The dietary energy intake (DEI) was 129.29 (117.57-133.89), 111.71 (100.42-133.47), 146.86 (128.03-163.18) kJ·kg-1·d-1 respectively. Nitrogen balance was positive in group A, B, C [2.99 (2.15-4.72) g, 1.20(0.59-1.89) g, 0.24 (-0.87-1.27) g]. The BUN decreased at the 7th and 10th day (P<0.01) in group C. The BUN and phosphorus in group A increased, but without significant difference as compared to baseline. No significant differences of nutritional status were found among three groups throughout the trial. ConclusionMinimal DPI 0.65 g·kg-1·d-1 plus the supplement of protein loss in dialysate can maintain the nitrogen balance in peritoneal dialysis patients.

Objective To investigate the clinical effect of arthroscopic anterior cruciate ligament(ACL) reconstruction using autologous multi-strands of hamstring tendon.Method From May 2002 to Dec.2006,arthroscopic reconstruction of the anterior cruciate ligament using multi-strands of hamstring tendon was performed on 39 patients.Among them,28 cases were followed up for 7 to 48 months(average 13.5±2.3 months).Result The median Lysholm knee score was improved from 47.3±3.6 to 91.3±2.9 after operation.Conclusion Arthruscopic reconstruction of the anterior cruciate ligament using muhi-strands of hamstring tendons is a good alternative method with minimal invasion and convenient tendon harvesting and fewer complication and reliable clinical effect.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Ascites ; pathology ; Biopsy ; Biopsy, Fine-Needle ; Child ; Cytodiagnosis ; methods ; Diagnosis, Differential ; Female ; Humans ; Leukemia-Lymphoma, Adult T-Cell ; pathology ; Lymphoma, B-Cell ; pathology ; Lymphoma, Large B-Cell, Diffuse ; pathology ; Lymphoma, T-Cell ; pathology ; Male ; Middle Aged ; Pleural Effusion ; pathology ; Young Adult