Objective To detect the levels of transforming growth factor-?_1(TGF-?_1) in plasma,serum and urinary of children with Immunoglobulin A glomerulonephritis(IgAGN) and mesangial proliferate glomerulonephritis(MsPGN) and explore the different effects of TGF-?_1 in the two diseases.Methods The plasma,serum and urinary TGF-?_1 levels were measured in 24 children with IgAGN,and 30 children with MsPGN and 30 healthy controls by enzyme-linked immunosorbent assay(ELISA).Results The TGF-?_1 levels in plasma,serum and urinary samples of IgAGN group were increased.The TGF-?_1 levels of IgAGN were significantly higher than those of MsPGN and heathy controls(P(0.05)).Conclusion It is showed that TGF-?_1 plays a diffenent role in IgAGN and MsPGN.

Objective To investigate the roles and significances of MMP-2 and MMP-9 in diffuse proliferative lupus nephritis by repeated renal biopsy.Methods Seventeen patients diagnosed by renal biopsy as WHO typeⅣlupus nephritis were analyzed by immunohistochemistry staining for MMP-2 and MMP-9. Double staining for MMP-2 and MT1-MMP,MMP-9 and CD68 were also performed.Patients had repeated renal biopsy after followed up for 2.5 years.The relationship between expressions of gelatinases and pathological activity index and clinical data were studied.Results MMP-2 immunoreactivity was detected in normal controls and was increased in diffuse proliferative lupus nephritis.MMP-9 staining,which was almost negative in normal giomeruli,was increased much more significantly in diffuse proliferative lupus nephritis. The immunoreactivity of MMP-2 and MMP-9 was positive in MT1-MMP staining and CD68-positive macrophages, respectively.The expression of MMP-2 and MMP-9 was reduced by 70% and 62% in 10 patients whose clinical condition was partially alleviated,while the expressions in 7 patients whose clinical condition was not alleviated,were only reduced by 27% and 32%.The staining for MMP-2 and MMP-9 were correlated with activity index of lupus nephritis and proteinuria.Conclusion Up-regulation of gelatinases expression in diffuse proliferate lupus nephritis is correlated to activity index of the disease.

[Summary] Graves'ophthalmopathy ( GO) is the most common extrathyroidal manifestation of Graves disease ( GD) . It is an organ-specific autoimmune disorder. The estimated incidence of GO in the general population is 16 women and 3 men per 100,000 population every year. Approximately 3%-5% of patients with GO are exposed to the potential of losing their sight and requiring optimized medical intervention. For the clinicians it is helpful to assess the degree of activity and severity of GO;both are important in deciding whether a patient requires intervention and which type of intervention is indicated. These interventions include management of hyperthyroidism, measures to relieve local symptoms, immunosuppressive therapies for active moderate and severe GO, and finally rehabilitative surgery for inactive patients.

Adult ; Aged ; Case-Control Studies ; Female ; Humans ; Male ; Middle Aged ; Myelodysplastic Syndromes ; immunology ; metabolism ; T-Lymphocytes, Regulatory ; metabolism ; Th17 Cells ; metabolism

OBJECTIVETo determine if Ganoderma polysaccharides can antagonize prostaglandin E2 (PGE2)-induced suppression of murine splenocyte interferongamma (IFN-gamma) and tumor necrosis factor alpha (TNF-alpha) mRNA expression.

METHODSMixed lymphocyte culture reaction was used as the experimental model. The expressions levels of IFN-gamma and TNF-alpha mRNA were measured by semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR).

RESULTSAfter the cultures were treated with PGE2 for 4 h, IFN-gamma mRNA expression was reduced as compared with the control, which was especially obvious when PGE2 concentrations exceeded 10 micromol/L (P<0.01). Ganoderma polysaccharides above 100 mg/L showed partial antagonistic effect against the inhibition of IFN-gamma by PGE2 at the fixed concentration of 20 micromol/L. Further studies indicated that PGE2 (20 micromol/L) impaired the expression of TNF-alpha mRNA after an 8-hour incubation and Ganoderma polysaccharides above 100 mg/L could partially antagonize this effect.

CONCLUSIONGanoderma polysaccharides can partially antagonize PGE2-induced suppression of murine splenocyte IFN-gamma and TNF-alpha mRNA expression.

Animals ; Cells, Cultured ; Dinoprostone ; pharmacology ; Female ; Gene Expression ; drug effects ; Interferon-gamma ; genetics ; Lymphocytes ; cytology ; drug effects ; metabolism ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Polysaccharides ; pharmacology ; RNA, Messenger ; biosynthesis ; genetics ; Reishi ; chemistry ; Reverse Transcriptase Polymerase Chain Reaction ; Spleen ; cytology ; Tumor Necrosis Factor-alpha ; genetics

OBJECTIVETo clarify the role of RhoC in the growth of hepatocellular carcinoma cells and its molecular mechanism, so as to explore the molecular target of tumor cell growth.

METHODSsiRNA-RhoC plasmid was constructed and RhoC gene silencing the cell-line of hepatocellular carcinoma was setup. Cell growth was assessed by MTT assay. AgNORs staining was applied to determine cell proliferation. Plate cell clone test was conducted to examine the capacity of cell clone formation. FACS was adopted to measure the course of cell cycle and semi-quantitative RT-PCR was used to determine the expression of cell cycle proteins. In order to further determine the effect of RhoC expression on cell growth, a RhoC over-expression human hepatocellular cell line was setup by PcDNA3-RhoC plasmid transfection.

RESULTSThe inhibition rate of RhoC was 82.3%. From the fourth day of cell culture, the growth of cells in RNAi group was significantly slower than that in parental Bel7402 and negative control groups (0.41 ± 0.10 vs. 0.73 ± 0.11 and 0.71 ± 0.07 respectively, P < 0.05). AgNORs staining showed that average cell stained particles in RNAi group was significantly lower than that in parental Bel7402 and negative control(1.23 ± 0.35 vs. 3.47 ± 0.93 and 3.17 ± 0.78, P < 0.01). Plate clone formation test showed that clone formation efficiency in the RNAi group was notably lower than that in the control group [(20.33 ± 5.42)% vs. (70.58 ± 10.10)% and (69.83 ± 14.77)%, respectively, P < 0.01]. Cell cycle analysis by FACS showed that G(0)/G(1) cell percentage in the RNAi group was significantly higher than that in the control group [(73.14 ± 5.93)% vs. (57.05 ± 5.97)% and (52.99 ± 4.80)%, P < 0.05]. Compared with Bel7402 and negative control groups, the expression of following growth associated genes was significantly decreased: cyclin D1(0.45 ± 0.21 vs. 1.25 ± 0.24 and 1.12 ± 0.15, respectively, P < 0.05)and CDK4 (0.55 ± 0.08 vs. 1.18 ± 0.32 and 1.10 ± 0.29, respectively, P < 0.05); the following genes were notably increased: p16(1.07 ± 0.23 vs. 0.36 ± 0.12 and 0.35 ± 0.13, respectively, P < 0.01)and p21(0.42 ± 0.12 vs. 0.17 ± 0.06 and 0.19 ± 0.08, respectively, P < 0.05). RhoC was highly expressed in PcDNA3-RhoC transfected hepatocellular cell line. From the third day on of the cell culture, cell growth in PcDNA3-RhoC group was remarkably higher than that in the HL7702 and PcDNA3 groups (0.83 ± 0.10 vs. 0.54 ± 0.11 and 0.58 ± 0.55, respectively, P < 0.05).

CONCLUSIONSRhoC is the key molecule in promoting hepatocellular cell growth, and is a promising target for tumor cell growth controlling.

Carcinoma, Hepatocellular ; genetics ; metabolism ; pathology ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; Cyclin D1 ; metabolism ; Cyclin-Dependent Kinase 4 ; metabolism ; Cyclin-Dependent Kinase Inhibitor p16 ; metabolism ; Cyclin-Dependent Kinase Inhibitor p21 ; metabolism ; Humans ; Liver Neoplasms ; genetics ; metabolism ; pathology ; Plasmids ; RNA Interference ; RNA, Small Interfering ; genetics ; Transfection ; rho GTP-Binding Proteins ; genetics ; metabolism ; rhoC GTP-Binding Protein

OBJECTIVETo study the antitumor effect of saponin extracted from Tupistra chinensis Baker (STCB) against mouse sarcoma S-180 cell proliferation in vitro and in vivo and explore the primary mechanism of this effect.

METHODSCytotoxic effect of STCB on S-180 cells in vitro was evaluated by MTT colorimetry, and its effect against in vitro tumor growth was tested in Kunmin mice bearing S-180 implanted tumor. The morphological and ultrastructural changes of S-180 cells after saponin treatment in vitro were examined with light and transmission electron microscope. Flow cytometry was performed to examine the cell cycle and apoptosis of S180 cells treated with different concentrations of STCB with propidium iodide staining.

RESULTSSTCB could markedly inhibit S-180 cell proliferation in vitro with 50% inhibitory concentration of 34.64 microg/ml. STCB given by intragastric administration also significantly inhibited the growth of S-180 solid tumor, and the inhibition rate exceeded 30% at the dose of 0.5 g/kg, reaching 54.86% at 2 g/kg. Electron microscopy and flow cytometry revealed increased S180 tumor cell apoptotic rate with the increment of saponin concentration, along with increased percentage of cells in S phase and decreased cells in G(2)/M phase in response to 10 or 30 microg/ml STCB treatment. At the concentration of 60 microg/ml, however, STCB resulted in an opposite effect on the cell cycles, presumably due to its interference with mitosis at high concentrations.

CONCLUSIONSSTCB inhibits the growth of S-180 cells both in vivo and in vitro possibly by inducing cell apoptosis and interfering with the cell cycle progression of the tumor cells.

Animals ; Antineoplastic Agents, Phytogenic ; pharmacology ; therapeutic use ; Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Liliaceae ; chemistry ; Male ; Mice ; Phytotherapy ; Saponins ; pharmacology ; therapeutic use ; Sarcoma 180 ; drug therapy ; pathology

OBJECTIVETo demonstrate the changes of PAI-1 activity and mRNA expressions induced by tumor necrosis factor alpha (TNF-alpha) or minimally modified low-density lipoprotein (mmLDL) in human umbilical vein endothelial cells (HUVECs).

METHODSThe PAI-1 activity in HUVECs culture medium was measured by chromogenic assay. The PAI-1 mRNA expressions were determined by Northern blot. The mitogen-activated protein kinase-kinase (MAPKK) inhibitor and protein kinase C (PKC) inhibitor were used to intervene the inductions by TNF-alpha or mmLDL.

RESULTSThe PAI-1 activity and mRNA level were increased when HUVECs were exposed to TNF-alpha or mmLDL. The MAPKK inhibitor (PD98059, 60 mumol/L) could markedly inhibit PAI-1 activity and mRNA expression induced by TNF-alpha (100 U/ml) or mmLDL (50 micrograms/ml). But Staurosporine (10 nmol/L) and H7 (15 mumol/L), which were the PKC inhibitors, had no significant inhibition effect.

CONCLUSIONS(1) TNF-alpha or mmLDL could induce PAI-1 activity and mRNA expression in HUVECs. (2) Increase of PAI-1 activity induced by TNF-alpha and mmLDL was related to its mRNA expression. (3) The MAPK pathway may play a role in PAI-1 expression induced by TNF-alpha or mmLDL.

Endothelium, Vascular ; cytology ; metabolism ; Female ; Humans ; Lipoproteins, LDL ; pharmacology ; Mitogen-Activated Protein Kinases ; metabolism ; Plasminogen Activator Inhibitor 1 ; genetics ; metabolism ; RNA, Messenger ; metabolism ; Tumor Necrosis Factor-alpha ; pharmacology ; Umbilical Veins ; cytology

OBJECTIVETo investigate the Epstein-Barr virus (EBV) infection, the expression of EBV latent membrane protein 1 ( LMPl) and oncogene bcl-2 in lung cancer patients.

METHODSEBERI in 108 cases of lung cancer were detected with in situ hybridization. EBV positive and negative lung cancer tissues were analysed for the expression of LMP1 and Bcl-2 by immnohistochemistry. The average area (AA) and integral optical density (IA) of each sample was measured with the digital medical image analyzing system.

RESULTSIn 108 cases of lung cancer, 36 cases were EBER1 positive and 7 cases were LMP1 positive. The expression of Bcl-2 was higher in EBV positive lung cancer tissues than that in EBV negative. The AA value was 58014.23 +/- 6918.45 and 38156.22 +/- 4096.79, while the IA value was 11.00 +/- 1.48 and 8.03 +/- 0.78 respectively. No statistic difference was fund in the expression of Bcl-2 betwen LMP1 positive and negative lung cancer tisssues.

CONCLUSIONEBV infection in lung cancer increased the expression of bcl-2, which may play a role in the occurrence or development of lung cancer. The increased expression of Bcl-2 may not be induced by LMP1. The exact mechanism need further study.

Adult ; Aged ; Epstein-Barr Virus Infections ; genetics ; metabolism ; pathology ; virology ; Gene Expression Regulation, Viral ; Herpesvirus 4, Human ; genetics ; physiology ; Humans ; Lung Neoplasms ; genetics ; metabolism ; pathology ; virology ; Male ; Middle Aged ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; Viral Matrix Proteins ; genetics ; metabolism

Objective To reconstruct a computational fluid dynamics (CFD) model of human nasal cavity, and make comparison analysis with acoustic rhinometry and rhinomanometry. Methods One healthy volunteer was performed CT scanning of nasal cavity, three dimensional CFD model was established by Simplant 10.0 and Gambit 2.3.16, and Fluent 6.3.2 was employed to simulate the airflow of nasal cavity. Acoustic rhinometer was used to assess the area of nasal cavity, rhinomanometry was adopted to measure the airflow and intranasal pressure drop during inspiration, and the results were compared with those obtained from CFD model. Results Cross section area of nasal cavity obtained from CFD model matches well with that measured by acoustic rhinometer within 30 mm distance from nostril, while the latter was larger than the former beyond 50 mm distance from nostril. The trend of intranasal pressure drop at different airflows measured by CFD model was the same as that measured by rhinomanometry, while the transnasal pressure obtained by CFD model was lower than that recorded by rhinomanometry. Conclusion CFD model can accurately simulate the shape of nasal cavity and measure the parameters of intranasal airflow, which helps to understand the airflow characteristics of nasal cavity.