BACKGROUND:The studies addressing osteoporosis in male starts relatively late,which lack effective and acknowledged method in treating.Epimedium,a Chinese medicine can be used in treating with osteoporosis for its obvious regulation of immune system,skeleton and reproductive system.OBJECTIVE:To investigate the effects of epimedium on bone mineral density(BMD) and bone structure performance in male castrated rats.DESIGN,TIME AND SETTING:The randomized controlled animal study was performed at the Geriatric Disease Laboratory of Tongji Medical College,Huazhong University of Science and Technology from January 2007 to April 2008.MATERIALS:Fifty adult,healthy,male,Sprague Dawley rats,weighing 340?10 g,with 15-week-old,were selected to prepare osteoporosis models.METHODS:Fifty rats were assigned to the control,model and three treated groups,with 10 animals in each group.The rats in the treated groups were treated by epimedium with different doses at 2 weeks after models preparation for 12 weeks.The doses were 1.0(low dose),5.0(medium dose),10.0(high dose) g/(kg?d) respectively.The animals in the other group were treated by aequales sodium chloride.MAIN OUTCOME MEASURES:Biochemical markers,BMD and bone morphometry of rats the fourth lumbar vertebra was evaluated after damaged by self-made SL22000 bone fatigue system.RESULTS:The BMD in the control,medium high dose groups were higher than that of the model group(P

As the code of behavior, honest-trust has its theoretical foundation of ethics. From the point of view of morality-justice, honest-trust is the ethical code of behavior; from the point of view of utilitarianism, honest-trust is the basic demand for increasing social welfare; from the point of view of responsibility and right, honest-trust is the responsibility and right of social members. Nowadays society should construct the social environment and mechanism of honest-trust, which can not be limited in the views of morality-justice and utilitarianism, but should be constructed in term of managing society and based on the responsibility and right.

Objective To investigate the value of sulindac in chemoprevention and treatment of human hepatocellular carcinoma. Methods The hepatocellular carcinoma cell(HCC) lines (SMMC 7721 and HepG 2) were used in this study. Antiproliferation effects was measured by MTT assay, and apoptosis, cyclooxygenase 2 (COX 2) and Bcl 2 were detected by Western dot blotting. Results The growth inhibition and apoptosis of HCC cells could be induced by sulindac in a dose and time dependent manner. The effects of growth inhibition and apoptosis induction were different between 2 cell lines. After 24 hours of incubation with sulindac at the concentration of 2 mmol/L and 4 mmol/L, the expression of COX 2 and Bcl 2 were markedly reduced. Conclusions Sulindac could inhibit the growth of two HCC cell lines effectively in vitro by induction of apoptosis, which were associated with the inhibition of COX 2 and Bcl 2 expression.

Objective To explore the correlation of tumor growth and endothelial progenitor cells (EPC) entering blood induced by surgical injury in tumor bearing nude mice. Methods Forty-two tumor bearing nude mice were randomly divided into seven groups (n=6): non-surgical injury groups (1 d and 30 d), anesthetic group, surgical injury groups (24 h, 48 h, 72 h and 30 d after surgery). Blood samples and xenograft tumor tissues were taken from anesthetic group 24 h after anaesthesia and surgical injury groups 24 h, 48 h, 72 h and 30 d after surgery. EPC levels in peripheral blood were measured by flow cytometry, serum VEGF levels were determined by ELISA, microvessel density (MVD) and expression of VEGF were detected by immunohistochemistry. Results The levels of EPC in 24 h post-surgery group, 48 h post-surgery group and 72 h post-surgery group were significantly higher than that in non-surgical injury 1 d group (P<0.05). The levels of VEGF in 24 h post-surgery group, 48 h post-surgery group, 72 h post-surgery group and anesthetic group were significantly higher than that in non-surgical injury 1 d group (P<0.05). There was no significant difference in MVD among groups (P>0.05). Pearson correlation analysis revealed that serum VEGF levels were related to EPC levels in peripheral blood (r=0.695 6, P<0.01), while EPC levels in peripheral blood were not related to MVD (r=0.221 4, P>0.05), and serum VEGF levels had no correlation with MVD (r=0.224 9, P>0.05). Conclusion Surgical injury has no obvious influence on xenograft tumor growth.

Objective To explore the relationship between the severity of experimental acute pancreatitis (AP) and the expression of Myc interacting zinc finger protein 1 (MIZ1) in rat, to evaluate the value of MIZ1 for severity assessment .Methods Acute necrotizing pancreatitis ( ANP) model was induced by retrograde injection of sodium taurocholate into the pancreatic duct , and normal saline was used in control group.The rats were sacrificed at 6, 24, 48 h, and the blood and pancreas tissue was collected , and serum amylase level , C reactive protein ( CRP ) , TNF-α, IL-6 and MIZ1 were determined by ELISA .Pancreatic tissue was routinely examined by pathologist , and the MIZ1 protein expression in pancreatic tissue was measured by immunohistochemistry and Western blot .Results The serum amylase levels of control group and ANP group at 6, 24, 48 h were (449 ±40), (578 ±25), (1 021 ±205), (971 ±143)U/L, and the levels of CRP were (123 ±23), (169 ±25), (226 ±34), (229 ±24)mg/L;and the levels of IL-6 were (20.16 ± 4.11), (38.60 ±12.05), (52.33 ±6.77), (44.83 ±4.30)ng/L;and the levels of TNF-αwere (55.33 ±3.32), (82.8 ±5.26), (120.66 ±16.00), (108.33 ±12.17)ng/L;and the levels of MIZ1 were (5.51 ± 0.34), (3.44 ±0.56), (2.11 ±0.11), (2.41 ±0.43) ng/L.The pathologic scores of pancreas were (1.83 ±0.75), (6.00 ±1.67), (8.16 ±2.70), (9.33 ±1.50), and the expressions of MIZ1 in pancreatic tissue were 0.81 ±0.05, 0.53 ±0.07, 0.31 ±0.06, 0.21 ±0.08.Except for amylase level of ANP 6h group, other parameters of ANP group were significantly different with those of control group , and the parameters of ANP 24, 48 h group were significantly different with that of ANP 6 h group ( P<0.01), but there was no significantly different between ANP 24 and 48 h group.MIZ1 expression was negatively correlated with serum amylase level , CRP, TNF-α, IL-6 and pathologic scores of pancreas , and the difference was statistically signific ant (P<0.01).Conclusions The decreasing expression of MIZ1 is closely correlated with the severity of AP , and may be a potential marker for prognosis evaluation .

[ ABSTRACT] AIM:To investigate the molecular mechanism and the immunosuppressive phenotype of macropha-ges under long-term exposure to lipopolysaccharide ( LPS) .METHODS:We used Ficoll-Hypaque density gradient centri-fugation combined with MicroBeads Separation Kits to separate peripheral blood mononuclear cells from human blood, and then induced the monocytes into macrophages.We observed the morphology of the macrophages by treating the cells with LPS for 48 h, in comparison with a negative control and IFN-γtreatment.ELISA was used to detect the levels of cytokines, such as IL-10, IL-12, IL-6 and TNF-α, and flow cytometry was used to detect the expression of the surface molecules (HLA-DR, CD14, CCR7, HLA-ABC and CD40).To observe the effect of macrophage on T cell proliferation, co-culture experiment was carried out for 6 d.Real-time PCR was used to validate the expression levels of molecules related to MyD88-independent pathway in Toll-like receptor 4 ( TLR4) signal pathway.RESULTS:The antigen-presenting ability of the macrophages was reduced and the IL-10 expression level was increased after the cells were treated with LPS for 48 h. We observed a poor proliferative capacity of CD8 +T cells after co-culturing of LPS-induced macrophages with CD3+T cells for 6 d.The results of real-time PCR indicated that TRIF, IRF3 and CIITA were down-regulated in LPS-induced macropha-ges.CONCLUSION:We successfully established a macrophage model in vitro and observed that LPS-induced macropha-ges into an immunosuppressive phenotype with poor CD8 +T cell proliferative capacity, in which MyD88-independent TLR4 signaling pathway was impaired.

Objective To study the effect of Smad4 gene on angiogenesis related factors in human gastric cancer cell line.Methods Recombinant eukaryotie expressing plasmid pcDNA3.1 (-)-Smad4 containing Smad4 gene and empty vector pcDNA3.1 (-) were introduced into human gastric cancer cell line MKN28 using lipofectam and selected by G418,respectively.Two cell lines were obtained as follows:Smad4+-MKN28 cell line which was MKN28 transfected with a stable hybrid containing Smad4 gene and Smad4--MKN28 cell line with empty plasmid as control.The transcription and expression of VEGF and TSP1 were investigated by RT-PCR and Western blot.Results The mRNA expression of TSP1 in Smad4+-MKN28 cells was higher (P＜0.05) than that in control cells,while VEGF was lower(P＜0.05).Western blot showed the consistent results as measurement by RT-PCR.Conclusion Smad4 restoration in gastric cancer cells reduced angiogenesis rates through down-regulation of angiogenesis activitor and up-regulation of angiogenesis inhibitor.

AIM: To discuss the effects of phytoestrogenic-genistein on cathepsin K (CK) expression stimulated by interleukin-1α (IL-1α) in osteoclast-like cells (OCLs) . METHODS: The OCLs were isolated from tissue of human giant cell tumor of bone (GCT) . The cells treated with reagents were divided into 7 groups including control (treated with phenol red-free-DMEM), vehicle (treated with 1.2 nmol· L-1 IL- 1α), 10-10-10-6genistein, genistein+ ICI 182.780, and 17[β-estrodiol (17β-E2) group. The cells were treated with 1.2 nmol· L-1IL-1α after pre-treated with genistein or 17β-E2 for 48 h (excluded the control group) . Expression of CK wasdetermined by reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blot in OCLs stimulated by IL-1α in the presence of genistein or 17[β-E2. RESULTS: The obvious increase of expression of CK by IL1α in vehicle group was noted in comparing with control group (P ＜ 0.01 ) . Genistein down-regulated CK gene expression stimulated by IL-1α at the transcription level in a dose-dependent manner (r = 0.68, P ＜ 0.01 ) .Genistein down-regulated CK protein expression stimulated by IL-1α also in a dose-dependent manner (r = 0.61,P ＜ 0.01 ). The effects of genistein were abrogated partly after treatment with the estrogen receptor antagonist ICI 182.780. CONCLUSION: Genistein inhibits CK expression stimulated by IL-1α partly through estrogen receptor in OCLs.

Objective To investigate the biological behavior and expression of intergrin-linked kinase (ILK) after exposure to transforming growth factor beta 2 (TGF-β32) in human Tenon fibroblasts (HTFs).Methods The primary ceils of HTFs were cultured by tissue attached culture method and identified by immunofluorescence analysis with Vimentin and keratin.The proliferation levels of HTFs induced by different concentrations of TGF-β2 were analyzed by MTT.The α-smooth muscle actin (α-SMA) and ILK mRNA expression were analyzed by quantitative Real-time PCR.The protein expression of α-SMA,ILK and E-cadherin were analyzed by Western Blot,and the protein expression of α-SMA and E-cadherin were also analyzed by immunofluorescence staining.Results MTT analysis showed that the optical density levels of 5.0 μg · L-1 and 10.0 μg · L-1 TGF-β2 were significantly higher than those of 0.1 μg · L 1,1.0 μg · L-1 and the control after exposure for 48 hours and 72 hours,and all these optical density levels were significantly higher than that for 24 hours (all P ＜0.05).The expression of α-SMA and ILK mRNA increased significantly when cells were treated with 5.0 μg · L-1 TGF-β2 for 48 hours in comparison with the control group (all P ＜ 0.05).The protein of α-SMA,ILK and E-cadherin were expressed both in TGF-β2 treated groups and control group,and TGF-β2 up-regulated the expression of them.There were significant differences when compared with the control group (all P ＜ 0.05).Immunofluorescent staining showed that α-SMA and E-cadherin were detected in TGF-β2 treated groups.α-SMA expressed in cytoplasm,while E-cadherin both in cytoplasm and nucleus.Conclusion TGF-β2 can induce the proliferation,transdifferentiation and adhesion of HTFs,and up-regulate the expression of ILK in vitro,suggests that ILK may play a role in the process of scar formation after glaucoma filtration surgery.

Objective To investigate the Mycobacterium tuberculosis infections in 2014 among Beijing army recruits, and evaluate a new method for screening latent tuberculosis infections.Methods A total of 194 army recruits were subjected to chest X-ray examination purified protein derivative(PPD) skin test, antibody detection, and interferon gamma release assay(IGRA) by ELISA combined with recombinant protein CFP10-ESAT6 and latent infection protein Rv2628.Results The positive rates of PPD skin test and antibody test were 49.7% and 15.5%, respectively.The latent infection rate of IGRA test was 22.2% in 194 cases after CFP10-ESAT6 stimulation.After stimulation of latent tuberculosis infection(LTBI) with Rv2628, IFN-γ level was significantly higher than that in healthy control group (P<0.05).The weak positive group of TST (5 mm≤diameter<15 mm) had a significantly higher level of IFN-γ than the strong positive group(diameter≥15 mm)(P<0.05),but after stimulation with CFP10-ESAT6,IFN-γ levels were not significantly different between the two groups(P>0.05).There was no significant difference between antibody negative and positive groups after stimulation by CFP10-ESAT6 and Rv2628 (P>0.05).The area under the ROC curve of Rv2628 diagnosis of tuberculosis infection was 0.84.When Youden index was 0.621,the specificity was 94.7% and sensitivity was 67.4%.ConclusionCombined detection of antigens Rv2628 and CFP10-ESAT6 specific IFN-γ values can be potentially used for differential diagnosis of active or latent tuberculosis infections.