Objective To investigate whether the hyper-reactivity of monocytes from the patients with primary biliary cirrhosis (PBC) to LPS and Poly I ∶ C was associated with miR-146a.Methods Peripheral blood mononuclear cells from PBC patients and healthy controls were stimulated with 10 μg/ml of LPS or Poly I ∶ C.The levels of IL-1 β,IL-6,TNF-α and IFN-α in the culture medium were detected by ELISA.The relative expressions of miR-146a,miR-21,let-7e,miR-155 and miR-125b were detected by RT-PCR.The regulatory role of miR-146a on the production of inflammatory factors in LPS or Poly I ∶ C stimulated THP-1 cells was studied by gain-and-loss function assay.Results The up-regulation of miR146a,which was induced by both LPS or Poly I ∶ C,was impaired in the monocytes from PBC patients.The production of LPS or Poly I ∶ C induced inflammatory factor,could be enhanced by miR-146a down-regulation.Conclusion The hyper-reactivity of monocytes from PBC patients to LPS and Poly I ∶ C was associated with miR-146a.

Objective To investigate the increased expression of microRNA-146a(miR-146a) in peripheral blood mononuclear cells (PBMC) of patients with chronic immune thrombocytopenic purpura (ITP) and its clinical significance. Methods Twenty-eight patients with chronic ITP and 28 healthy controls matched with age and gender were enrolled in this study. Fluorescent quantitative PCR reaction was used to detect the relative expression of miR-146a in their PBMC. The serum concentration of TNF-α, IL-2,IL-1 β and IFN-γ were measured by ELISA. CCK-8 method was used to detect the proliferation ability of PBMC , which transfected with miR-146a mimics or inhibitor and then stimulated with platelet . Results The relative expression of miR-146a in ITP patients was higher than that of healthy controls. The increased expression of miR-146a was negatively correlated with the serum TNF-α, IL-2 and IFN-γ. The PBMC transfected with miR-146a mimics had reduced expression of IL-2 and proliferation when stimulated with platelet.In contrast, the opposite effect was observed with the miR-146a inhibitors transfection. Conclusion MiR146a was involved in the pathogenesis of chronic ITP by controlling IL-2 production and PBMC proliferation.Thus, it may be a potential therapy target for chronic ITP.

Objective： To investigate the effect of c ytokines (IFN-γ，TNF and IL-1) on the expression of CD40 and CD40 ligand (CD4 0L) in monocytes/macrophages. Methods： The mRNA expression of C D40 and CD40L was measured by RT-PCR and the CD40，CD40L expression on the mono cytes/macrophages were detected by flow cytometric analysis. Results： IFN-γ，TNF and IL-1 could not only significantly up-regulate the mRNA levels of CD40 and CD40L in cultured monocytes/macrophages, but also increase t he expression of CD40 and CD40L. Antioxidant VitE could reduce the expression o f CD40 and CD40L induced by IFN-γ，TNF and IL-1. Conclusion： IFN-γ，TNF and IL-1 can stimulate high expression of CD40 and CD40L . Antio xidant VitE can partially inhibit the expression of CD40 and CD40L induced by cy tokines in cultured monocytes/macrophages.

Objective: To determine the eff ect of probucol on adhesion of human monocytic line THP-1 induced by oxidized low density lipoprotein (oxLDL). Methods: THP-1 cells were induced by oxLDL in vitro. The CD11b, CD54 expressions and adhesion to human umbilic al vein endothelial cells (HUVEC) were measured after treatment with probucol at different concentrations by flow cytometry and β-nitrophenyl N-acetyl-β-D -glucosminide test. Results: Probucol inhibited the adhesion of oxLDL-induced THP-1 cells to HUVEC and down regulated the expression of CD11b in a dose dependent manner (P<0.01), but there was no inhibition on exp ression of CD54. Conclusion: Probucol can inhibit adhesion and a ggregation of monocyte-macrophages to endothelium in circulation, and may have anti-inflammatory action.

Objective To observe the long-term effect of surgical treatment in infertile varicocele patients. Methods 180 infertile patients with varicocele accepted modified Palomo operation (retroperitoneal high ligation of internal spermatic cord vein) from 2004 to 2007, and the long-term effect of thesurgery were analyzed retrospectively, such as semen parameters and spouse pregnancy. Results The operations were performed successfully in 180 cases, and semen parameters improved significantly after the surgery (Ps ＜ 0. 05 ).Among all patients, 10 cases had spouse pregnancy within 3 months after operation;47 cases had spouse pregnancy from 3 to 6 months after operation; 25 cases had spouse pregnancy from 6 to 12 months after operation;13 cases had spouse pregnancy from 12 to 24 months after operation. The fertility rate was 52. 8% (95/180) in total. Conclusion Surgical treatment of infertile male patients with varicocele has good long-term effects, made it the first choice in these patients.

Objective To detect the expression of IL-27 in PBC (primary biliary cirrhosis)patients and the possible involvement of IL-27 signal pathway in PBC. Methods The gene transcription and protein expression levels of IL-27 in patients with PBC, chronic hepatitis B(CHB) group and healthy controls(HC) were measured by real-time PCR, ELISA, flow cytometry and immunohistochemistry. AST,ALP, ALT, TBIL, GGT were determined and their correlation with IL-27 was also analyzed. Results IL-27 was significantly elevated in patients with PBC and IL-27 is present in the liver tissues of patients with PBC. Expression of IL-27 on CD4+T cells was increased in patients with PBC(72.40% ±6.22% ) and CHB(59.40% ± 7.03%) compared with HC(1.70%±0.55%,P＜0.01). Expression of IL-27 protein was increased in patients with PBC [( 126.25 ± 36.00 ) pg/ml] compared with CHB [( 51.81 ± 23.30 )pg/ml, P ＜ 0. 01] and HC[(34.19 ± 9.70) pg/ml, P ＜ 0.01], and it was positively correlated with GGT( r = 0.554, P＜0.01) and TBIL (r = 0.559,P＜0.01), but no correlation with ALT, AST, ALP.Conclusion These facts indicated the key role of IL-27 in the immune inflammatory reaction in patients with PBC.

Objective To investigate the association between IL-22 and the pathogenesis of coronary artery atherosclerosis(AS).Methods The relative expression of IL-22 mRNA in PBMC from 30 AS patients and 8 patients without any signs of coronary artery stenosis was detected by RT-PCR.Serum IL-22 levels of 22 patients without any signs of coronary artery stenosis and 79 AS patients were detected by ELISA.CRL-1730 cells(human umbilical vein endothelial cells) were stimulated with oxidized low density lipoprotein (ox-LDL) at different dosage for 24 h,and the expression of IL-22R1 was detected by flowcytometry.The proliferation ability of CRL-1730 cells treated with IL-22(20 ng/ml) and/or ox-LDL(100 μg/ml)was measured by MTS assay,and the expression of basic fibroblast growth factor(bFGF) was detected by RTPCR and ELISA.Results Decreased IL-22 expression in PBMC and serum was observed as worsen of AS.The expression of IL-22R1 in ox-LDL treated CRL-1730 cells was increased in dose dependent manner.OxLDL decreased proliferation ability,as well as bFGF expression and releasing,of CRL-1730 cells.This effect of ox-LDL was partially rescued by IL-22.Conclusion IL-22 may have anti-atherosclerosis effect.This effect may be mediated by regulating bFGF expression and endothelial cells proliferation ability in the presence of IL-22.

Adiponectin is an adipokine predominantly synthesized and secreted by adipocytes in the white adipose tissue, and it can lower the blood glucose level and increase free fatty acid oxidation. In the current study, we developed the globular domain of adiponectin (gapM1) to treat type II diabetes. In both flask and fermentor, we cultivated Pichia pastoris expressing recombinant gapM1 and established the purification procedure by using gel filtration and anion exchange chromatography. To evaluate the biological activity of recombinant gapM1, we used rat type II diabetes model fed high-fat diet in combination with low-dose STZ (Streptozocin) induction. We purified 200 mg gapM1 with purity of 96% from 10 liters of supernatant. The recombinant gapM1 significantly lowered blood glucose (34.2%), serum triglyceride (79.6%) and total cholesterol (62.1%) in type II diabetes induced rat. Therefore, the recombinant human gapM1 is successfully expressed in Pichia pastoris and effectively treated type II diabetes in rat models.
Adiponectin ; biosynthesis ; genetics ; pharmacology ; therapeutic use ; Animals ; Diabetes Mellitus, Experimental ; drug therapy ; Genetic Vectors ; Hypoglycemic Agents ; pharmacology ; therapeutic use ; Male ; Pichia ; genetics ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Recombinant Proteins ; biosynthesis ; genetics ; pharmacology ; therapeutic use

OBJECTIVETo investigate clinical implications of expression of CD40L in monocytes and changes in serum soluble CD40L in patients with acute coronary syndromes (ACS).

METHODSSixteen control and 56 patients, including 24 with stable angina (SA), 20 with unstable angina (UA) and 12 with acute myocardial infarction (AMI) enrolled in this study. Expression of CD40L in monocytes was analyzed by flow cytometry and sCD40L levels were measured by ELISA.

RESULTSExpression of CD40L in monocytes and serum levels of sCD40L in UA and AMI patients were higher than in SA patients and controls. In patients with AMI, sCD40L levels showed no significant increase when compared to patients with UA, while AMI patients had a peak level of sCD40L at 24 hours after AMI. PTCA induced a marked rise in sCD40L levels in all patients, while CD40L expression in monocytes showed no difference between patients with PTCA, before and after.

CONCLUSIONEnhanced level of serum sCD40L may be a reliable prognostic indicator for ACS and may represent a marker of coronary disease activity.

Angina Pectoris ; blood ; pathology ; CD40 Ligand ; blood ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Male ; Monocytes ; metabolism ; Myocardial Infarction ; blood ; pathology

OBJECTIVETo observe sperm motion parameters in pre-freeze and post-thaw semen samples using computer-assisted sperm analysis (CASA) system.

METHODSSemen analyses of 238 samples before freezing and after thawing were separately performed by Hamilton-Thorne Sperm Analyzer.

RESULTSSperm motility in post-thaw samples was significantly decreased. There was significant correlation and difference between pre-freeze and post-thaw samples in sperm motion parameters, including average path velocity (VAP), straight line velocity (VSL), curvilinear velocity (VCL), amplitude of lateral head displacement (ALH), straightness (STR) and linearity (LIN), except heat cross frequency (BCF). The percentage of sperm movement velocity parameters (VAP, VSL and VCL) and moving pattern parameters (ALH) significantly decreased, while that of LIN and STR significantly increased in post-thaw samples.

CONCLUSIONCASA system is of clinically applied value and is a useful tool for evaluating sperm motion parameters in pre-freeze and post-thaw semen samples.

Cryopreservation ; Diagnosis, Computer-Assisted ; Freezing ; Humans ; Male ; Sperm Motility