Objective To investigate the correlations of plasma eotaxin-2,soluble tumor necrosis factor receptor(sTNFR) Ⅱ and other cytokines levels with the status of metabolic syndrome(MS) in type 2 diabetic nephropathy (DN) patients treated with maintenance hemodialysis(MHD).Methods Thirty type 2 DN patients with stable pathogenetic conditions and MHD treatment for more than 3 months,thirty newly diagnosed and untreated type 2 diabetes(T2D) patients and thirty healthy volunteers were enrolled in the study.The MS related markers,including blood pressure,waist circumference,body mass index,triglyceride(TG),high density lipoprotein cholesterol(HDL-C),fasting blood glucose (FBG),total cholesterol(TC),low density lipoprotein cholesterol(LDL-C) and C reactive protein(CRP),were determined.The expression levels of 40 cytokines were detected with the AAH-INF-G3 antibody microarray,and their correlations with MS related markers were analyzed by the Pearson method.Results The incidence rates of MS in DN patients with MHD and T2D patients were 88.89％ and 33.33％,respectively.Compared with T2D patients and healthy controls,the DN patients had more MS related markers,higher waist circumference and lower body mass index,and their plasma eotaxin-2,I-309 and sTNFR Ⅰ / Ⅱ levels increased significantly.Pearson correlation analysis showed that plasma eotaxin-2 and I-309 levels were positively related to MS risk factors such as TG,FBG and diastolic blood pressure(DBP) levels.Plasma sTNFR Ⅱ and I-309 levels were significantly positively correlated with plasma CRP levels.Conclusion A micro inflammatory state exists in type 2 DN patients with MHD.Abnormal glycolipid metabolism may influence on the immunologic and physiological state of human body,and then form the micro inflammatory state.Eotaxin-2 and I-309 may participate in this chronic and persistent process and further induce renal damage by upregulating sTNFR Ⅰ / Ⅱ levels,which may result in the formation of MS state in type 2 DN patients with MHD.

Regional microenviroment hypoxia is a common feature in inflammation and malignancy. Cells are able to trigger an adaptive response to hypoxia conditions that result in hypoxia-inducible factor(HIF)overexpression. Based on a brief description of the structure and functions of HIF, this article discusses the relationship between inflammation, tumor and HIF.

Objective:To investigate the condition of B cell differentiation from cord blood CD34+CD19-hematopoietic stem/progenitor cells in vitro.Methods:CD34+CD19-cells from cord blood were isolated and purified by using immunomagnetic beads separation system.CD34+CD19-cells supported by murine S-17 stromal cells were stimulated in co-culture with T3 and cytokines.Differentiated B cells were analyzed by flow cytometry.Results:The amplification of the B cells derived from CD34+CD19-hematopoietic stem/progenitor cells in co-culture with T3 and IL-7 reached 198-fold of increase,most of the induced cells expressed CD10 and CD19.Conclusion:In the experimental conditions selected,co-culture of CD34+CD19-cells with T3,IL-7 and murine S-17 stromal cells could stimulate differentiate toward to B cells in vitro.

Objectives To establish a reliable method for isolating and purifying soluble collagen type Ⅱ(SCⅡ)by optimizing preparative procedure.Method The chicken sternal cartilage was selected as raw material.Guanidine hydrochloride was used to remove the proteoglycans.The digestion manners of pepsin,sodium chloride concentrations for salting,types of DEAE anion resin were studied for extracting SCⅡ.The SCⅡ identification was made by SDS-PAGE,absorption spectrum and amino acid analysis.Result It was convenient for pre-treatments of chicken sternal cartilage.The proteoglycans could be efficiently removed by 4 mol/L guanidine hydrochloride.The satisfied results were obtained by limited enzyme digestion of pepsin added by two steps.The optimizing concentration of sodium chloride for salting was 2.4mol/L.SDS-PAGE maps revealed that the bands of purified SCⅡand standard CⅡ were at the same location.The absorption peak of SCⅡ was at 230nm.The concentrations of Gly,Pro and Ala were the highest in 15 amino acids.Conclusion The improved method has significant advantages of simple working process,result reliability and convenient source of raw material.It is suitable for purifying the SCⅡ at variable scales in research works and clinic application.The SCⅡ product obtained has high purity and accords with the characteristics of collagen type Ⅱ.

Objective To study the alteration of Th17 cells and related molecules in patients with Hashimoto's thyroiditis (HT).Methods Th17 cells were determined by flow cytometry.Real-time PCR method was applied to detect the expression of the orphan nuclear receptor RORγt and interleukin(IL) -17.Serum IL-6 and IL-23 were detected by ELISA method.Results ( 1 ) Compared with healthy controls,the frequency of Th17 cells [ (0.75± 0.79) ％ vs ( 0.28 ± 0.23 )％ ] and expression of RORγt ( 0.30 ± 0.38 vs 0.04 ± 0.02,both P ＜ 0.05 ) were significantly increased in HT patients.( 2 ) The levels of IL-6 and IL23 in HT patients were higher than those in healthy controls ( 3.66 ± 4.70 vs 0.47 ± 1.11,154.7 ± 75.81 vs 80.65 ± 61.41,both P＜0.05 ).( 3 ) A positive correlation between Th17 cells and serum TgAb was revealed in HT patients ( r =0.848 4,P =0.007 7 ).(4) The results of PCR showed that IL-17 and RORγt expressed in thyroid tissues of patients HT.Conclusion An increased frequency of Thl7 cells was found in HT patients,implying that this cell subset may play an important role in the pathogenesis of autoimmune thyroid disease.

Objective:To explore the mechanism of B10 cell involved in cardiomyocyte hypertrophy following myocarditis, and to develop potential therapeutic strategies.Methods:BALB/c mice infected with Coxsackie virus B3 induced viral myocarditis model. The expression of angiotensin (ANG)Ⅱ and its receptor in myocarditis mice was detected. The changes of B10 cells in the hearts of control mice and myocarditis mice were analyzed by flow cytometry. After losartan was administered to myocarditis mice, the degree of myocardial inflammation was detected by HE staining, the expression of inflammatory factors was detected by ELISA, the myocardial hypertrophy was detected by wheat germ agglutinin (WGA) staining, and the changes of B10 cells in the heart were analyzed by flow cytometry. The levels of cardiac troponin T (C-TNT) and high mobility group box 1 (HMGB1) protein in neonatal mouse cardiomyocytes treated with ANGⅡ and ANGⅡ+ IL-10 were detected. Cardiomyocytes were treated with ANGⅡ, ANGⅡ+ B10 cells, ANGⅡ+ B10 cells + IL-10 receptor antibody and ANGⅡ+ B cells to detect C-TNT protein levels, and Annexin-V/PI was used to detect the apoptosis of cardiomyocytes. Cardiomyocytes were treated with oxidized HMGB1, reduced HMGB1 and disulfide HMGB1, and C-TNT expression was detected.Results:Coxsackievirus B3 infection caused cardiac hypertrophy, high expression of ANGⅡ and its receptor, and transient increase of B10 cells in mice. Losartan treatment blocked the angiotensin receptor, reduced expansion of B10 cells. B10 cells alleviated apoptosis of cardiomyocytes and inhibited the production of HMGB1 induced by ANGⅡ patch by producing IL-10, thus alleviating viral myocarditis and cardiac hypertrophy.Conclusions:B10 cells may play an important role in myocardial protection in myocarditis.

ObjectiveTo understand the pathological and physiological roles of Presenilin 1 (PS1) in Alzheimer's disease (AD) recurrence, and the interaction between PSI and carhoxyl terminus of Hsc70 interacting protein (CHIP). MethodsThe yeast two-hybrid system was applied to identify a novel PS1 interacting protein as CHIP. After pGBKT7-PS1-C203 bait plasmid and full fragement CHIP of pACT2-CHIP expression vector were constructed, the interaction between PSI and CHIP was tested by β-galactosidase assay, pGBKT7-PS1-C203 was co-transfected with pACT2-CHIP into 293T cells and the interaction between PS1 and CHIP was tested by co-immunoprecipitation and Western blot. ResultsSpecificity of the interaction between PS1 and CHIP was identified by β-galactosidase assay and co- immunoprecipitation. ConclusionsCHIP is able to modulate chaperone functions and the pathway of protein ubiquitination/degradation. CHIP may regulate a proper assembly of the γ-secretase complex through its interaction with PSI, which is helpful to elucidate the mechanism of AD pathology.

Objective To investigate the role of interleukin-17 (IL-17) in spinal dorsal horns in neuropathic pain (NP) in rats and its effect on activation of astrocytes.Methods In vivo experiment Sixty-four male SPF Sprague-Dawley rats,aged 6-8 weeks,weighing 180-200 g,were randomly divided into 3 groups using a random number table:control group (group C,n =16),sham operation group (group S,n =24) and group NP (n =24).The animals were anesthetized with intraperitoneal pentobarbital sodium,the L5,6 spinal nerves of the left side of the rat were gently separated and exposed,tightly ligated with 5-0 silk suture and transected.In group S,the L5,6 spinal nerves of the left side of the rat were only exposed.In group C,no operation was performed.Mechanical pain threshold was measured at day 1 before operation and days 1,3,5,7,10 and 14 after operation.The expression of IL-17,IL-6,IL-1β and tumor necrosis factor-alpha (TNF-α) mRNA in the spinal dorsal horn was determined using quantitative real-time PCR at day 7 and day 14 after operation.At day 7 after operation,the activation of astrocytes in the spinal dorsal horn was detected.In vitro experiment Primarily cultured astrocytes of neonatal rats were randomly divided into 4 groups using a random number table:control group (group C,n=22),10 ng/ml IL-17 group (I10 group,n=18),50 ng/ml IL-17 group (I50 group,n-18) and 100 ng/ml IL-17 group (I100 group,n=22).In I10,I50 and I100 groups,the astrocytes were incubated with the culture medium containing 10,50 and 100 ng/ml IL-17,respectively.The proliferation of astrocytes was detected by MTT at 24,48 and 72 h of incutation or culture.The expression of IL-6,IL-1β and TNF-α mRNA was determined using quantitative real-time PCR.Results In vivo experiment Compared with group C,the mechanical pain threshold was significantly decreased at 3-14 days after operation,the expression of IL-17,IL-6 and IL-1β mRNA in the spinal dorsal horn was up-regualted at 7 days after operation,and the activation of astrocytes was increased in group NP,and no significant change was detected in the mechanical pain threshold at each time point after operation in group S.In vitro experiment Compared with group C,the proliferation of astrocytes was significantly increased at 48 h of incubation in I10 and I50 groups,the proliferation of astrocytes was significantly increased at 48 and 72 h of incubation,and the expression of IL-6 and IL-1β mRNA was up-regulated in I100 group,and no significant change was found in the proliferation of astrocytes in group S.Conclusion Up-regulated expression of IL-17 in spinal dorsal horns may be involved in the maintenance of NP,and the mechanism is related to promoted activation of astrocytes and induced inflammatory responses in rats.

AIM: To examine T-lymphocyte rDNA transcription activity in peripheral blood of patients with gastrointestinal maliganant tumor and to clarify its clinical significance.METHODS: T-lymphocyte rDNA transcription activity in peripheral blood of 48 cases of patients with stomach-intestine tract malignant tumor were measured.RESULTS: Before surgery, the T-lymphocyte rDNA transcviption activity was obviously lower than that after surgery, also lower than that of the normal control( P

Objective To investigate the prevalence of genes conferring aminoglycoside resistance in multidrug-resistant strains of Acinetobacter baumannii (MDR-ABA).Methods Multidrug-resistant A.baumannii strains were isolated during the period from August to November 2012 from patients in the affiliated hospital of Jiangsu University and the First Hospital of Zhen-jiang.Kirby-Bauer diffusion method was used to determine the susceptibility of these strains to antimicrobial agents.PCR was performed to detect the aminoglycoside resistance genes.Results The 36 MDR-ABA strains showed high resistance rates to most antimicrobial agents except cefoperazone-sulbactam.The prevalence of the genes conferring aminoglycoside resistance, aac (3)-I,aac (6’)-Ib,aph (3’)-I and armA,was 72.2% (26/36),72.2% (26/36),80.6% (29/36)and 80.6% (29/36), respectively.Conclusions The MDR-ABA strains in this study are highly resistant to antimicrobial agents,which is closely as-sociated with the genes conferring aminoglycoside resistance.