Objective To observe the changes of extracellular matrix(ECM) production and hypertrophy of glomerular mesangial cells(GMCs) induced by high glucose(HG),and the inhibitory role of myriocin(ISP-1).Methods GMC cultured with normal glucose(5.6 mmol/L D-Glucose,NG),HG(25.2 mmol/L D-Glucose) and HG plus ISP-1(100 mg/L) for different durations(0,24,48,(72 h)).The sizes of GMC were indicated by forward scatter intensity,measured by flow cytometery,and the levels of fibronection(FN),collagen Ⅳ(Col Ⅳ),laminin(LN),precollagen Ⅲ(Pcol Ⅲ) and hyaluronic acid(HA) in the supernatant of cultured GMC were detec-(ted) by ELISA.Results Compared with NG,HG could induce GMC hypertrophy(P

Objective - - To analyze the prevalence and influencing factors of multi site work related musculoskeletal disorders （ ） Methods WMSDs in surgeons. A total of 102 surgeons from four hospitals were selected as study subjects by convenient sampling method. The Chinese version of Musculoskeletal Disorders Questionnaire was used to investigate the prevalence of ， Results WMSDs in the past one year the related individuals and occupational factors. The total prevalence of WMSDs among （ ）， （ ） （ ） surgeons was 54.9%. The top three sites were neck 48.0% lower back 35.3% and shoulder 32.4% . The prevalence of （ vs ，P ） WMSDs in multiple sites was higher than that in a single site 43.1% 11.8% <0.01 . Multivariate logistic regression ， ， analysis showed that surgeons who smoked were tired at work and had a bent back had a higher risk of developing WMSDs ［ （ - ）， （ - ）， （ - ）， P ］ odds ratios and 95% confidence intervals were 3.66 1.41 9.46 8.33 2.15 32.20 and 18.74 2.14 166.77 all <0.01 Conclusion - after excluding the influence of confounding factors. The prevalence rate of multi site WMSDs among surgeons is ， high and the influencing factors include bad living habits and occupational factors such as working load and working posture.

OBJECTIVE This study aimed to investigate the influence of IgD on T/B cell activationand construct hIgD-Fc-Ig fusion protein to competitive inhibition IgD binding with IgDR. METHODS T/B cells were sorted by magnetic cell sorting. The differences of mIgD and IgD-R level between different T/B cell subtypes were detected by FCM. Serum IgD level was detected by ELISA. Human IgD-Fc-IgG1- Fc sequence was amplified by cross- PCR and then subcloned into PET28a(+ ) empty vector. After prokaryotic expression through escherichia coli, we obtained the hIgD-Fc-Ig fusion protein by affinity chromatograph. Western blot was used to identify the hIgD- Fc- Ig fusion protein. Human peripheral blood monouclear cells (PBMC) and fibroblast like synoviocytes (FLS) proliferation were detected using a cell counting kit-8 (CCK-8). RESULTS The percentage of CD3+/CD4+, CD3+/IgD+, CD3+/CD4+/IgD+, CD3+/IgD-R+ and CD3+/CD4+/IgD-R+ cells increased significantly in RA patients comparing to healthy people. IgD can stimulate PBMC proliferation. IgD (1, 3, 10, 30 μg·mL-1) stimulate PBMC proliferation significantly after 24 h. We obtained stable and active hIgD-Fc-Ig fusion protein. The hIgD-Fc-Ig fusion protein showed no effect on PBMC proliferation. But it could downregulate human IgD protein promoting proliferation effects in human PBMC. CONCLUSION This result suggests that IgD and IgDR play an important role on T/B cell activation in RA patients and the hIgD-Fc-Ig fusion protein may competitively inhibit IgD's function and may play an therapeutic role in autoimmune diseases.

OBJECTIVEMyriocin (ISP-1) is a new type of immune inhibitor extracted from cordyceps sinensis. This study was to observe the effects of ISP-1 on the expression of cell cycle regulatory protein D1 (cyclinD1) in high glucose-induced hypertrophy rat glomerular mesangial cells (GMCs).

METHODSRat GMCs were cultured in vitro and divided into three groups: high glucose (450 mg/dL D-glucose), normal glucose (100 mg/dL D-glucose, control) and ISP-1 (450 mg/dL D-glucose plus 100 μg/mL ISP-1). The protein expression of cyclinD1 was detected by flow cytometry.

RESULTSThe expression of cyclinD1 in GMCs in the high glucose group increased significantly in a time-dependent manner compared with that in the control group. ISP-1 treatment significantly inhibited the up-regulated expression of cyclinD1 induced by high concentration glucose, and the expression of cyclinD1 was restored to the level of the control group 48 and 72 hrs after ISP-1 treatment.

CONCLUSIONSHigh concentration of glucose can up-regulate the expression of cyclinD1 in GMCs. ISP-1 may inhibit the up-regulated expression of cyclinD1, which might contribute to the protective effect of ISP-1 against GMC hypertrophy induced by high glucose.

Animals ; Cyclin D1 ; analysis ; Fatty Acids, Monounsaturated ; pharmacology ; G1 Phase ; Glucose ; toxicity ; Hypertrophy ; Mesangial Cells ; chemistry ; pathology ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo study the influences of endocannabinoid-anandamide (AEA) on the proliferation and apoptosis of the colorectal cancer cell line (CaCo-2) and to elucidate the effects of CB1 and lipid rafts, and to further elucidate the molecular mechanism and the effect of AEA on the generation and development of colorectal cancer.

METHODSHuman colorectal cancer cell line CaCo-2 was cultured in RPMI 1640 medium supplemented with 10% fetal bovine serum in 5% CO(2) atmosphere at 37°C. CaCo-2 cells were divided into different groups and treated with different concentrations of AEA, AEA + SR141716A, AEA + AM630 and AEA + methyl-β-cyclodextrin (MCD). MTT assay was used to determine the effects of AEA, its putative CB1, CB2 receptor antagonists (SR141716A and AM630) and MCD on the proliferation of CaCo-2 cells. Annexin V-PE/7AAD binding assay was used to detect apoptosis in the CaCo-2 cells. Western-blot was applied to check the expressions of CB1, CB2, p-AKT and caspase-3 proteins in different groups of CaCo-2 cells.

RESULTSAEA inhibited the proliferation of CaCo-2 cells in a concentration-dependent manner and the effect could be antagonized by SR141716A and MCD. The inhibiting rates were (21.52 ± 0.45)%, (42.16 ± 0.21)%, (73.64 ± 0.73)% and (83.28 ± 0.71)%, respectively, at different concentrations of AEA (5, 10, 20 and 40 µmol/L). The three groups (20 µmol/L AEA, 20 µmol/L AEA + 10 µmol/L SR141716A and 20 µmol/L AEA + 1 mmol/L MCD) showed different inhibiting rates [(73.64 ± 0.73)%, (16.15 ± 0.75)% and (12.58 ± 0.63)%], respectively. Annexin V-PE/7AAD binding assay showed that AEA induced apoptosis in the CaCo-2 cells and MCD could antagonize this effect. The apoptosis rates of the three groups (control, 20 µmol/L AEA and 20 µmol/L AEA + 1 mmol/L MCD) were (2.95 ± 0.73)%, (39.61 ± 0.73)% and (14.10 ± 0.64)%, respectively. The expressions of CB1, CB2, p-AKT and Caspase-3 proteins were all observed in the CaCo-2 cells. AEA inhibited p-AKT protein expression and induced caspase-3 protein expression. The two actions were also antagonized by MCD.

CONCLUSIONSAEA can strongly suppress the proliferation of colorectal cancer CaCo-2 cells via the CB1 receptor and membrane cholesterol-LRs and induce apoptosis via lipid rafts. Anandamide plays a very important role in the carcinogenesis and development of colorectal cancer. MCD is a critical member in this system.

Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Arachidonic Acids ; antagonists & inhibitors ; pharmacology ; Caco-2 Cells ; Cannabinoid Receptor Modulators ; antagonists & inhibitors ; pharmacology ; Caspase 3 ; metabolism ; Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Endocannabinoids ; Humans ; Indoles ; pharmacology ; Membrane Microdomains ; metabolism ; Piperidines ; pharmacology ; Polyunsaturated Alkamides ; antagonists & inhibitors ; pharmacology ; Proto-Oncogene Proteins c-akt ; metabolism ; Pyrazoles ; pharmacology ; Receptor, Cannabinoid, CB1 ; antagonists & inhibitors ; metabolism ; Receptor, Cannabinoid, CB2 ; antagonists & inhibitors ; metabolism ; beta-Cyclodextrins ; metabolism

OBJECTIVEHepatitis B virus-associated glomerulonephritis (HBV-GN) is an immune complex-mediated glomerulonephritis. The present study was conducted to identify HBV S gene mutation in children with HBV-GN.

METHODSSerum HBV DNA was extracted in 53 children, including 30 with HBV-GN, 5 with HBV-carrying nephrosis (control group 1), and 18 HBV carriers (control group 2). HBV S gene sequence was amplified by polymerase chain reaction (PCR). The PCR products were sequenced directly and compared with AY167097.1, an epidemic HBV strain in China.

RESULTS(1) The adw serotype of HBV was found in all the 30 cases with HBV-GN, 5 cases with HBV-carrying nephrosis and 17 HBV carriers except for 1, in whom adr serotype was identified. (2) HBV genotype B was found in 29 children with HBV-GN, 5 cases with HBV-carrying nephrosis and 17 HBV carriers, genotype E was found in a child with HBV-GN, and genotype C in an HBV carrier. (3) A total of 17 kinds of different single nucleotide change in HBV S gene were identified in 21 of 30 (70%) HBV-GN patients. Among them, 16 of 21 (76.2%) nucleotide mutations resulted in amino acid substitution. It was interesting that most (11/16, 68.8%) amino acid substitutions involved threonine, serine and tyrosine, the potential phosphorylation sites of mitogen-activated protein kinase (MAPK) and protein tyrosine kinase (PTK) in HBV protein. Single nucleotide changes which didn not result in amino acid substitution were found in 2 HBV-carrying nephrosis patients, 2 HBV carriers and 5 cases with HBV-GN.

CONCLUSIONSingle nucleotide changes in HBV S gene were found in most children with HBV-GN. Most mutations in HBsAg resulted in amino acid substitutions involving threonine, serine and tyrosine, which may play a role in the pathogenesis of HBV-GN.

Amino Acid Substitution ; Carrier State ; Child ; DNA Mutational Analysis ; DNA, Viral ; genetics ; Genes ; Genotype ; Glomerulonephritis ; virology ; Hepatitis B virus ; genetics ; Humans ; Mutation ; Viral Envelope Proteins ; genetics

OBJECTIVE To observe whether human CD4 + T cells could be activated by immuno-globulin D (IgD) via IgD receptor(IgDR)-Lck. METHODS Human CD4+ T cells were purified from peripheral blood mononuclear cells (PBMCs) with microbeads. The viability of T cells were detected by CCK-8. The binding affinity and expression of IgDR on T cells were detected by flow cytometry. The protein expression of IgDR, Lck and P-Lck were analyzed by western blot. RESULTS IgD could concentration-dependent bind to IgDR on CD4+ T cells. The expression of IgDR was increased in response to treatment with IgD in a time- dependent and concentration- dependent manner. Stimulating by IgD resulted in enhanced phosphorylation of Lck compared with that in the medium control sample. The expression of Lck was not changed. As inhibitor of PTK, Herbimycin A or A770041, which combined with IgD could significantly inhibit phosphorylation of Lck(Tyr394). The proliferation promoting effect of IgD was blocked by Herbimycin A or A770041. IgD could stimulate CD4+ T cell activation and proliferation through upreg?ulating activating tyrosine residue of Lck (Tyr394) phosphorylation. CONCLUSION These results demon?strate that IgD exaggerates CD4+T cell activities, which may be through promoting Lck phosphorylation.

OBJECTIVEMast cells and eosinophil have been found to play important roles not only in the development of anaphylactic inflammation but also in the chronic progression of organ reconstruction. But their role in the pathogenesis of Henoch-Schonlein purpura nephritis (HSPN) has not been fully understood. The present study was conducted to observe the serum levels of eosinophil cationic protein (ECP) and renal infiltration of mast cells in HSPN in order to elucidate their role in the development and progression of HSPN in children.

METHODSThe serum ECP levels were determined in 46 children with HSPN by fluoro-enzyme immunoassay (FEIA) using the Pharmacia CAP System. The distribution of mast cells infiltration was detected by immuno-enzyme-histological staining of tryptase (a marker for mast cell activation) and their relation with pathological changes was analyzed in 32 children with HSPN.

RESULTSThe serum ECP levels were 16.3 +/- 6.5 microg/L in the active stage of HSPN, significantly higher than that in remission stage (3.9 +/- 1.4 microg/L, P < 0.01) and that in control group (3.1 +/- 1.7 microg/L, P < 0.01). The number of mast cells in renal interstitium was 4.4 +/- 2.4 cells/mm2 in normal kidney, and significantly increased to 27.2 +/- 19.2 cells/mm2 in children with HSPN ISKDC grade II (P < 0.01) and 42.1 +/- 16.4 cells/mm2 in grade III (P < 0.05 when compared with grade II), 77.9 +/- 15.0 cells/mm2 in grade IV (P < 0.05 when compared with grade III).

CONCLUSIONThe serum ECP level could reflect disease activity of HSPN, and mast cell infiltration in kidney correlated significantly with renal histological severity in HSPN. Mast cells and eosinophil may play important roles in the development and progression of HSPN.

Biomarkers ; blood ; Child ; Disease Progression ; Eosinophil Cationic Protein ; blood ; Female ; Fluoroimmunoassay ; Humans ; Immunohistochemistry ; Kidney ; metabolism ; pathology ; Male ; Mast Cells ; metabolism ; pathology ; Nephritis ; blood ; metabolism ; pathology ; Purpura, Schoenlein-Henoch ; blood ; metabolism ; pathology ; Severity of Illness Index ; Tryptases ; metabolism

Two hundreds postmenopausal women with osteopenia, aged 45-80, were randomly divided into 4 groups, and received different drug interventions. After treatment for 12 months, the lumbar spine bone mineral density(BMD)and total hip BMD in alendronate group increased significantly(3.5% and 2.6%, both P＜0.05), the lumbar spine BMD raised 2.0% in selective estrogen receptor regulator group(P＜0.05). Bone turnover indices improved in both groups(all P＜0.05). No change in BMD or bone turnover indices was found in patients treated with active vitamin D3and calcium.

Objective To investigate the risk factors for postmenopausal osteoporosis in Shanghai.Methods Five hundred postmenopausal community-dwelling women aged 45-80 years were recruited. The case-control study was performed from June 2008 to September 2008.A total number of 250 women with postmenopausal osteoporosis identified with their bone mineral density (BMD) were selected as case group, and 250 non-osteoporosis women were selected as control group. BMD was measured by dual energy X-ray absorption (DEXA). Results Univariate logistic regression analysis showed that age, eduction level, occupation, years since menopause, BMI, use of calcium,historyofnon-violencefracture,fall,diabetesmellitus,chronicgastricdiseases, gastrointestinal resection and diarrhea were related to osteoporosis.Multiple logistic regression showed that age, years since menopause and nutritional status were the risk factors for osteoporosis. ConclusionsThe occurrence of osteoporosis is related with many factors in postmenopausal women in Shanghai, and women with early menopause, low BMI and older age should pay more attention to the prevention and treatment of osteoporosis.