Correlation between CD39+ and CD73+ regulatory T cells and liver injury of primary biliary cholangitis / 临床检验杂志

Objective To evaluate the changes of adenosine metabolism pathway related molecules and their contribution to inflammatory injury in primary biliary cholangitis (PBC).Methods The consecutive samples of 49 subjects with PBC from The First People's Hospital of Taicang and The Second People's Hospital of Changshu were recruited from October 2016 to October 2017,and 36 healthy controls were involved in this study.The expression of CD39 and CD73 on CD4+T cells and Foxp3 + regulatory T cells were assayed by flow cytometry and the concentration of adenosine triphosphate (ATP) in serum was analyzed by ultraperformance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF/MS).The correlations between Tregs,ATP and liver function were analyzed,i.e.,alanine aminotransferase (ALT),aspartate aminotransferase (AST),gamma-glutamyltransferase (GGT),alkaline phosphatase (ALP) and Mayo scores.Results In the patients with PBC,low proportions of CD4+CD39+T cells were noted compared with healthy controls [(5.28 ± 1.92) ％ vs (11.0l ± 3.19) ％,t =10.25,P ＜ 0.01].The patients with PBC also had significantly low proportion of CD4+CD25 + Foxp3+ CD39+ T cells compared with healthy controls [(23.75 ± 9.48) ％ vs (54.68 ± 5.18) ％,t =13.79,P ＜0.01].No significant difference of the proportion of CD4+CD73+T or CD4+CD25+ Foxp3+CD39+T cells was found between PBC and control groups (t values were 2.235 and 1.083,P ＞ 0.05).The level of serum ATP was higher in the patients with PBC than that of healthy controls [(200.28 ± 79.41) μg/L vs (89.20 ± 33.76) μg/L,t =8.367,P ＜ 0.01].A significant correlation was demonstrated between the proportion of CD39 + Treg in total Treg cells and the levels of ATP (r =-0.413,P =0.003),GGT (r=-0.378,P=0.007) and Mayo score (r=-0.382,P=0.007).Conclusion The low proportion of CD39+ Treg cells may contribute to the down-regulation of ATP hydrolysis in the patients with PBC.No significant change of CD73 + Treg cells was found in PBC patients.

Analysis and functional study on lncRNA expression profile in primary biliary cirrhosis patients / 中华检验医学杂志

Objective Analyzing the lncRNA expression profile in peripheral blood mononuclear cells(PBMC)of primary biliary cirrhosis(PBC)patients to provide new ideas for the pathogenesis,clinical diagnosis and treatment of PBC.Methods Collected peripheral blood from 30 PBC patients and 30 healthy volunteers, then separated PBMC.Four cases from each group were selected for long-noncoding RNA (lncRNA)expression microarray detection.Reverse transcription-PCR technology in a larger sample size was used to verify the microarray results.Bioinformatic analysis such as Cis-/Trans-target genes Gene ontology(GO)and pathway analysis, co-expression networks were conducted in order to provide a theoretical basis in the pathogenesis of PBC.Transfecting small interfering RNAs(siRNAs)for linc-pbc to see changes in the expression of nuclear receptor 4A group 3(NR4A3)and forkhead boxP3(FOXP3)and cell apoptosis in transfected PBMC.Results Compared to the healthy group, 749 lncRNA and 230 coding messenger RNA(mRNA)genes were abnormally expressed.Interestingly,NR4A3 gene was down-regulated by 78%.While linc-pbc, which was about 20 000 bp downstream of NR4A3 gene, increased 2.56-fold. Then design siRNA for linc-pbc.After transfection, mRNA and protein levels of NR4A3 and FOXP3 were up-regulated.Conclusions By recruiting PRC2 complex, linc-pbc may increased the methylation level of NR4A3 gene promoter region,thus decreasing the expression of NR4A3 in PBC patients and reduced NR4A3 futher downregulated the expression of FOXP 3 and reduced the amounts of immunosuppressive Treg cells in peripheral blood and liver tissue, breaking the equilibrium state of immune tolerance, and promoted the occurrence and development of the disease.

Establishment of methods to evaluate the quality of two different separate gel tubes during ISO15189 accreditation / 国际检验医学杂志

Objective To explore the establishment of methods to evaluate the quality of two different separate gel tubes .Meth-ods An evaluation comparing two separate gel tubes with silicate glass tubes ,which are the standard tubes ,was performed using data from 50 subjects .The serum samples were assayed for routine chemistry or immunology after centrifugation using the quantita-tive or qualitative detection methods such as ECL ,enzyme-linked immunosorbent assay (ELISA) ,reflection luminescence or immu-nofluorescence .The data were collect for further statistical analysis .Results For quantitative results of the items ,the two separate gel tubes ,compared with the standard glass tube ,there was no significant difference between the test results (P>0 .05);for quali-tative projects ,two separate plastic tube ,compared with the standard glass tube ,really the positive rate and the true negative rate was no significant difference (P>0 .05 ,K>0 .75) .Conclusion By using different detection methods and test items ,it is demon-strated that using two separate gel tube for items assay does not affect the accuracy and consistency of the test results .As a result , the method for external supplies quality assessment is established in the laboratory for ISO15189 quality control .

Prognostic prediction of primary biliary cirrhosis and therapy monitoring with ursodeoxycholic acid / 中华检验医学杂志

Primary biliary cirrhosis (PBC) is a chronic cholestic autoimmune liver disease and ursodeoxycholic acid (UDCA) is the first-line drug for the treatment.Currently there are several standards applied for the prognostic and therapeutic predictions of PBC,as well as for the guidance of personalized treatment.This paper elucidated the prognostic predicting factors in PBC and their applications in therapy monitoring with UDCA.Current challenges and future research interests are also put forward.

Clinical applications of long non-coding RNA detection in cancers / 中华检验医学杂志

In recent years, the discovery of thousands of long non-coding RNAs ( lncRNAs) has certainly changed human′s view of the complexity of mammalian genomes and transcriptome, as they play an important role in the regulation of transcription, post-transcription as well as epigenetic level, widely involved in various physiological process and diseases.Here this review summarized the lncRNAs associated with cancers, discussed the established methods used to detect and quantify lncRNAs, and evaluated the clinical application of lncRNAs as biomarker.

New era in autoantibodies detection: quantitation / 中华检验医学杂志

Autoantibodies are useful laboratory parameters for diagnosis of autoimmune diseases (AID).Methods for the detection of autoantibodies have achieved quantitative or semi-quantitative to some extent.Furthermore,with the development of detection technology,high-throughout and quantitative technology has been the trend in autoantibody measurement.Compared with qualitative results,the quantitative ones may provide more values for diagnosis,prediction,prognosis and therapeutic monitoring for AID.Therefore,although quantitative technology of autoantibodies is still faced a number of challenges,the detection for autoantibodies has come into the era of quantitation.

Expression of IL-10+CD19+ regulatory B cells in peripheral blood of patients with chronic hepatitis B inaction / 中华检验医学杂志

Objective To investigate the population and role of IL-10+ CD19+ regulatory B cell (Breg) in patients with chronic hepatitis B.Methods Patients with acute hepatitis B (AHB) (n =28),chronic hepatitis B (CHB) (n =31) and normal subjects (n =25) were collected from Changshu No.2 People's Hospital between 2011 June and 2012 October.Peripheral blood mononuclear cells (PBMC) were isolated and stimulated with CpG ODN 2006 and PMA.Flow cytometry was used to analyze the population of IL-10-CD19 + Breg,CD4 + CD25high Treg,and ELISA was used to analyze the concentration of IL-10 in culture supernatant.Results The population of Breg in Peripheral blood of the CHB group [1.28％ (1.05％-2.20％)] was higher than that in the AHB group [0.87％(0.55％-1.22％)] and the HCs group [0.89％ (0.51％-1.37％)] (P =0.001,0.006),and the difference between the AHB group and the HCs group was not statistically significant (P=0.669).Breg in the CHB group [14.30％ (10.70％-16.70％)] was higher than that in the AHB group [10.30％ (7.05％-13.30％)] and the HCs group [10.40％(6.85％-12.60％)] (P =0.003,0.001),treg in the CHB group [5.80％ (4.20％-9.10％)] was also higher than that in the AHB group [4.05％ (2.53％-5.40％)] and the HCs group [4.50％ (2.55％-5.50％)] (P ＜0.001,P =0.005),and there was no significantly difference between the AHB group and the HCs group (Breg:P =0.796 ; Treg:P =0.227).Spearman correlation analysis showed that Breg was positively correlated with Treg in the CHB group (r =0.50,P =0.004),however there was no significantly correlation in the AHB group and the HCs group (r =-0.15,P =0.462; r =0.09,P =0.669).The concentration of IL-10 in the CHB group was higher than that in the AHB group and the HCs group (P ＜ 0.001),and the difference between the AHB group and the HCs group was not statistically significant (P=0.341).Spearman correlation analysis showed that IL-10 were positively correlated with the population of Breg in the CHB group (r =0.409,P =0.022).Conclusion The poluations of regulatory B cell and regulatory T cell increased in patients with chronic hepatitis B,and Breg cell might play the immune regulation role through secreting IL-10 in chronic HBV infection.

miR-146a regulates monocyte hyper-reactivity to LPS and Poly I : C in patients with primary biliary cirrhosis / 中华微生物学和免疫学杂志

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.

Apolipoprotein C3 enhanced THP-1 cell adhesion to aortas of mice / 中华微生物学和免疫学杂志

Objective To investigate the enhancing effect of apolipoprotein C3 (APOC3) on THP-1 cell adhesion to aortas of mice.Methods Microsurgery was performed to separate the aorta of C57BL/6 mice in sterile condition.After stimulated by APOC3 (100 △g/ml) in vitro for 16 h,the aorta was allowed to adhere for 1 h with CFSE labeled THP-1 cells (1 ×106/ml).Then the adhesion effect was observed,and the expressions of vascular adhesion molecule 1 (VCAM-1) and intercellular adhesion molecule 1 (ICAM-1) were detected by immunohistochemical method.Results Adhesion effect of the mice aorta with THP-1 cells in the APOC3 stimulated group was stronger than the control group.Both the expressions of VCAM-1 and ICAM-1 in aortas were increased by APOC3,but the former was significantly up-regulated than the latter.Conclusion Apolipoprotein C3 could enhance THP-1 cell adhesion to aortas of mice.

The expression of sialic acid-binding immunoglobulin-like lectin-1 on peripheral blood mononuclear cells and its relationship with disease activity in rheumatoid arthritis / 中华风湿病学杂志

Objective To investigate the expression of sialic acid-binding immunoglobulin-like lectin-1 (Siglec-1) in the peripheral blood mononuclear cells (PBMCs) in patients with rheumatoid arthritis (RA),osteoarthritis (OA) and healthy controls and to explore the relationship between Siglec-1 expression and disease activity in RA.Methods Siglec-1 protein and mRNA levels were measured by flow cytometry and real-time quantitative reversetranscription-polymerase chain reaction (qRT-PCR) in 42 RA patients,28 OA patients and 26 healthy controls,respectively.The correlation studies between Siglec-1 and disease activity score 28 (DAS28) or C-reactive protein were performed.T-test was used for comparisons between groups and Pearson's correlation test was used for correlation analysis.Results The percentage of Siglec-1 positive cells of PBMCs in RA group [(15.2±7.6)％] was significantly higher than that in the OA group [(2.3 ±2.6)％] or healthy controls [(2.1±1.6)％,t=8.615,8.661; all P＜0.01].And the major cell type in PBMCs that expressed Siglec-1 was monocytes.The relative Siglec-1 mRNA expression in PBMCs in the RA group (3.4±1.5) was also significantly higher than that in the OA group (1.2±0.4) or healthy controls [(1.0± 0.4),t=3.446,3.966; all P＜0.05].But no significant differences of Siglec-1 protein and mRNA between the OA group and healthy controls were found.Furthermore,positive correlations between Siglec-1 protein and DAS28 or hs-CRP were found in RA patients (r=0.89,P＜0.01; r=0.48,P＜0.01).Conclusion PBMCs are activated which are characterized by elevated expression of Siglec-1 in RA patients.Circulating Siglec-1 may be considered as a potential noninvasive biomarker for monitoring disease activity in RA.

The role of microRNAs in the pathogenesis of autoimmune disease as systemic lupus erythematosus and rheumatoid arthritis / 中华检验医学杂志

MicroRNAs (miRNAs) are a new class of noncoding RNA which regulate gene expression at post-transcription level.It is believed that miRNAs are widely involved in the pathological and physiological processes,including the proliferation,development,differentiation and apoptosis regulation.

The role of monocytes Siglec-1 in stimulating CD4 + and CD8 + T cells proliferation and activation in atherosclerosis / 中华检验医学杂志

Objective To investigate the role of atherosclerotic monocytes Siglec-1 in stimulating CD4 + and CD8 + T lymphocytes proliferation and activation.Methods Experimental research.Cluster of differentiation antigen 14 (CD14) positive monocytes of 18 acute coronary syndrome (ACS),41 stable angina (SA) and 32 healthy volunteers were separated by magnetic-activated cell sorting.Different concentration of interferon-α (IFN-α,0,2,5,10 ng/ml) were used to up-regulate Siglec-1 and small interfering RNA (siRNA) or blocking antibody were used to down-regulate Siglec-1.Then monocytes were cocultured with CD4 + T/CD8 + T cells from a third healthy volunteer for 5 days.The experiment was designed for 11 groups (n=10 for each group),that was (1) normal CD14,(2) normal CD14 + IFN-α 5 ng/ml,(3) normal CD14 + IFN-α 5 ng/ml + anti-Siglec-1 2 μg/ml,(4) ACS CD14,(5) ACS CD14 + control siRNA group (Mock),(6) ACS CD14 + siRNA 679 40 nmol/L,(7) ACS CD14 + anti-Siglec-1 2 μg/ml,(8) SA CD14,(9) SA CD14 + Mock,(10) SA CD14 + siRNA 679 40 nmol/L and (11) SA CD14 + antiSiglec-1 2 μg/ml.Cell Counting Kit-8 (CCK-8) was used to determine T cells proliferation and ELISA was used to detect Interleukin-2 (IL-2),IL-10,IL-12 and IFN-γ of culture supematant.Data of cytokines detection were expressed as medium (quartiles) and analyzed by nonparametric rank sum test.Results By the blockage of Siglec-1 (group 6),the proliferation of CD4 and CD8 were decreased.Secretion of IL-2,IL-12 and IFN-γ by CD4 cells [67.00(62.50-87.30),0.86(0-1.63),and 47.82(37.60-56.67) pg/ml,respectively] were decreased and IL-10 [56.00(46.25-67.40) pg/ml] was increased compared with those in control group [group 4,213.70 (187.50-275.30),6.87 (4.90-8.93),114.90 (89.50-167.40),and 21.08 (15.70-33.20) pg/ml,respectively,with U =8.50,17.00,8.50,and 87.50,respectively.all P ＜ 0.05].When monocytes Siglec-1 from control group was up-regulated by IFN-α (group 2),secretion of IL-2,IL-12 and IFN-γ [220.44(174.30-312.30),7.90(6.540-10.40) and 143.75(78.20-210.00) pg/ml,respectively] were increased and IL-10 [21.95 (16.30-25.00) pg/ml] was decreased (compared with group 1,with U =89.50,98.00,100.00,and 0,respectively.all P ＜ 0.05).Regulation of Siglec-1 had no role in cytokines production in cocultured CD8 + T cells (all P ＞ 0.05).Conclusions IFN-α can upregulate monocytes Siglec-1.Siglec-1 may participate in the pathogenesis of AS via promoting proliferation of CD4 +/CD8 + T cells and Thl cytokines secretion of CD4 + T cells.

The progress of pathogenesis of autoimmune diseases / 中华检验医学杂志

Autoimmune diseases (AID) are a group of diseases,in which the tolerance of immune system to self component is broken.However,the etiology and pathogenesis of AID has not yet been clear so far.For better understanding the pathogenesis of AIDs and providing new idea on the diagnosis and treatment of AID,this review will focus on the latest development on pathogenesis of AID,including genetic background,environment factors,abnormal immune regulation,and the role of target cells.

MiR-146a regulate the proliferation and interleukin-2 production of T helpers from primary biliary cirrhosis patients / 中华风湿病学杂志

Objective To investigate the effect of miR-146a on the proliferation and interleukin (IL)-2 production of T helper cells from primary biliary cirrhosis (PBC) patients.Methods MiR-146a in the peripheral blood mononuclear cells (PBMC),monocytes,T helper cells,cytotoxic T cells and B cells from 20 confirmede PBC patients and age/sex matched healthy controls were detected by quantitative PCR.By gainand-loss of function,the miR-146a's effect on anti-CD3/anti-CD28 activated T helper's proliferation and IL-2 production ability were measured by CCK-8 approach and enzyme linked immunosorbent assays (ELISA),respectively.Statistical analysis were carried out by t-test.Results PBMCs (0.46±0.20 vs 1.00±0.26; t=7.47,P＜0.01),T helpers (0.33±0.13 vs 1.00±0.14; t=6.15,P＜0.01) and monocytes (0.56±0.11 vs 1.00±0.11; t=4.97,P＜0.05),but not B cells (0.91±0.06 vs 1.00±0.14; t=0.97,P＞0.05) and cytotoxic T cells (0.98±0.15vs 1.00±0.12; t=0.22; P＞0.05) from PBC patients had lower miR-146a expression level than that of healthy controls.Inducible up expression of miR-146a was observed in PBC patients'T helpers stimulated with antiCD3/anti-CD28 (1.00±0.18 vs 9.12±2.05; t=8.81; P＜0.01).The activated T helpers from PBC patients had higher proliferative ability [PBC:0.35±0.06 (A); healthy controls:0.26±0.04 (A); t=2.83; P＜0.05] and increased IL-2 production [PBC: (685.60±109.19 pg/ml)]; Healthy controls: [(512.20±72.26) pg/ml; t=2.96; P＜0.05 ] than those of healthy controls.For activated T helpers,the proliferation ability,as well as IL-2 production,was enhanced by miR-146a.Conclusion MiR-146a can down regulate the proliferation and IL-2 releasing of activated T helpers.The reduced miR-146a expression enhances IL-2 production and promotes proliferation of T helper of PBC patients,thus,may be involved in the pathogenesis of PBC.

Proliferation and activation induced cdl death of CD4 + T cells in childhood asthma / 国际儿科学杂志

Objective To investigate the correlation between immune inflammation and overactivity of T helper cells in childhood asthma by cell proliferation assay and activation induced cell death in vitro.Methods Th1/Th2/Th17 cytokines were determined by cytometric bead array.Cell proliferation and activation induced cell death were detected when CD4+ T cells were purified by magnetic beads and stimulated by PHA and antiCD3.At last,mRNA of Fas,FasL and Bcl-2 were mesured by real-time PCR.Results Cytokines of IL-4(2.451± 1.052ng/L vs 1.796 ±0.615 ng/L,P =0.018),IL-10( 1.920 ±0.813ng/L vs 1.390 ±0.162ng/L,P =0.006)and TNF(5.112 ±5.842 ng/L vs 1.506 ±0.551 ng/L,P =0.009) in sera of asthma group were higher than those in control group.Compared to control group,proliferation ability of CD4 + T cells in asthma group was greater ( OD450:0.498 ± 0.052 vs 0.274 ± 0.032,P ＜ 0.001 ) and apoptosis rate was lower( 35.62 ± 0.05 ％ vs 65.28±3.85％,P ＜0.001 ).mRNA expression of Fas in asthma group was lower but Bcl-2 was higher than those in control group.Conclusion It is implicated that defective expression of Fas and over expression of Bcl-2 in CD4+ T cells may contribute to apoptosis inhibition and cell proliferation,which could explain overeactivity of CD4 + T cells and lvmphocvte infiltration in childhood asthma.

IL-22 is involved in atherosclerosis lesions by regulated impaired proliferation ability of oxidized low density lipoprotein treated CRL-1730 / 中华微生物学和免疫学杂志

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.

Construction of lentiviral vector containing siRNA sequence of Siglec-1 and verification of inhibition efficiency / 中华微生物学和免疫学杂志

Objective To construct lentiviral vectors containing small interfering RNA (siRNA) sequence of Siglec-1 and to screen the effective vector.Methods Three fragments of Siglec-1 siRNA were designed and cloned into pGCSIL-GFP lentiviral plasmid.And then the plasmid was cotransfected into 293T cells with pHelper 1.0 and pHelper 2.0 plasmids.Forty-eight hours later,culture supernatant with virus particles was collected and concentrated.Virus titer was determined by 10-fold serial dilution method and virus was transduced into primary cultured mouse bone marrow-derived macrophages (BMM).Flow cytometry and QRT-PCR were used to screen effective vector with inhibition ability.Results Three vshRNA lentiviral plasmids and a control plasmid were constructed successfully and verified by DNA sequencing.Virus titer was between 1×10s TU/ml and 1×109 TU/ml,which was suitable for in vitro and in vivo experiments.The Lv-1 could inhibit Siglec-1 expression effectively in vitro transduction of BMM.Conclusion Lentiviral vectors containing siRNA sequence of Siglec-1 were constructed successfully and an effective vector was screened,which may lay the foundation for using the vector in gene knockdown experiment in vivo.

Expression of cytokines and suppressor of cytokine signaling in peripheral blood mononuclear cells derived dendritic cells from primary biliary cirrhosis patients / 中华检验医学杂志

Objective To study the role of suppressor of cytokine signaling ( SOCS ) in the pathogenesis of primary biliary cirrhosis( PBC),the levels of SOCS protein and the changes of function of dendritic cell(DC) were respectively observed from PBC patients.MethodsThe study population consisted of 10 patients of PBC and 8 healthy controls.Phenotypic analysis of cultured peripheral blood mononuclear cells (PBMC)-derived DC was performed by flow cytometry (FCM),such as CD83,CD86 and human leukocyte antigen DR (HLA-DR).The levels of interleukin-10 (IL-10),interferon-γ( IFN-γ) and IL-12 in culture supematant of DC were measured by enzyme linked immunosorbent assay ( ELISA ).The protein levels of SOCS1 and SOCS3 were detected by Western blot ( WB ).The features of changes in these parameters were analyzed between the two groups.ResultsThe expression of CD83,CD86 and HLA-DR in PBC patients were ( 79.4 ± 4.8 ) ％,( 86.5 ± 6.3 ) ％ and (90.0 ± 3.5 ) ％,which were significantly higher than those in healthy control group[ (68.3 ±4.1 )％,(74.2 ±6.3)％ and (83.6 ±7.6)％ ],respectively (t =5.340,4.120,2.514,P ＜0.05).The levels of IL-12 and IFN-γin PBC patients were (53.5 ± 11.1)and (32.0 ±9.0) ng/L,which were significantly higher than those in healthy control group[ (32.1 ± 10.7) and (15.4 ± 8.1 ) ng/L; t =4.123,3.818,P ＜ 0.01 ].There were not any significant difference of IL-10 level between PBC patients [ (7.0 ± 4.6) ng/L ] and the healthy controls [ ( 5.8 ± 4.2) ng/L; t =0.563,P ＞ 0.05 ].The proteins levels of SOCS1 and SOCS3 in PBMC-derived DC from PBC group were decreased significantly than those in healthy control group.ConclusionsThe results suggest that the PBMC-derived DC in PBC patients has greater ability of potent maturation and antigen presentation function.The decreased expression of SOCS levels may be associated with the excessive immunological reaction and the breakdown of self-tolerance.

Reform of laboratory diagnostics teaching mode for students of clinical medicine / 中华医学教育探索杂志

Laboratory diagnostics is an important discipline functioning as a bridge between basic and clinical medicine and it is closely associated with the diagnosis of clinical physician.But there are some problems in the laboratory diagnostic teaching including unreasonable curriculum standard,simple teaching method and unpractical theory.This paper explored and summarized the problems and the reform of laboratory diagnostics teaching mode for students of clinical medicine.

Increased expression of microRNA-146a in peripheral blood mononuclear cells of patients with chronic immune thrombocytopenic purpura and its clinical significance / 中华微生物学和免疫学杂志

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.