ABSTRACT
Objective To investigate the diagnosis and surgical therapy of delayed diaphragmatic rupture.Methods Forty patients with traumatic diaphragmatic rupture with delayed presentation were collected in Peking Uniom Medical College Hospital from January 2000 to December 2016.In all 40 patients, 36 patients had traumatic past history, 32 patients had clini-cal manifestations when diagnosed.Left-sided diaphragmatic rupture was found in 32 patients and right in 8 patients.1 patient received emergency surgery and 39 received selective surgery.38 patients received transthoracic surgery and 2 patients received combined thoracic-abdominal surgery.36 patients received direct diaphragm suture and 4 patients received patch repair.Re-sults All patients were recovered from the hospital.The median length of postoperative hospital stay was 11 days( range, 5-26 days).1 patient was found intestinal obstruction and received enterolysis 19 days after surgery.Conclusion Delayed traumat-ic diaphragmatic rupture is a rare but serious disease.Careful past history, physical examination and CT scan with reconstruc-tion of diaphragm are helpful in diagnosis and differential diagnosis.Surgical therapy after diagnosis is the best treatment.
ABSTRACT
Objective To isolate and identify exosomes from serum samples of the patients with polymyositis / dermatomyositis (PM/ DM),and analyze their protein composition preliminarily.Methods Exosomes from serum samples of the patients with PM/DM were isolated and purified by the ExoQuickTM kit.The morphological characteristics and particle size of exosomes were determined by transmission electron microscope (TEM) and NanoSight analyzer,respectively.The surface markers of exosomes such as CD9,CD81 and Flotillin-2 were identified by western blot.The concentration and composition of exosome protein were determined by the BCA method and SDS-PAGE,respectively.Results The exosomes from serum samples of PM/DM patients displayed round or oval vesicles with membrane structure under TEM,and their diameter range was about (92 ± 67) nm.western blot showed that these exosomes expressed CD9,CD81 and Flotillin-2.The total protein concentrations of exosomes in the patients with PM/DM and healthy controls were 14.68 (6.00,32.55) μg/μL and 14.09 (8.00,23.28) μg/μL,respectively.SDS-PAGE showed that high-abundance proteins enriched in 55-70 kD in both PM/DM patients and healthy controls,and that there were different bands in 40-55 kD between them.Conclusion Exosomes are isolated from serum samples of the patients with PM/DM successfully,and their protein concentration and composition are analyzed preliminarily,which provides the experimental evidences for further finding differential proteins.
ABSTRACT
Objective To evaluate the early diagnostic value of circulating microRNA-1 (miR-1) on acute myocardial infarction (AMI). Methods A prospective cohort study was conducted. The patients with chest pain admitted to the Second People's Hospital of Wuxi from November 2012 to June 2015 were enrolled. According to AMI diagnostic criteria, the patients were divided into AMI group and non-AMI group, and healthy individuals during the same period were served as heath controls. The venous samples of the onset patients were collected within 3 hours after admission. The plasma miR-1 was determined by real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR), and the levels of plasma cardiac troponin I (cTnI) and MB isoenzyme of creatine kinase (CK-MB) were measured by electrochemiluminescence. The correlation between plasma miR-1 and cTnI as well as CK-MB was performed by Spearman analysis. The early diagnostic performance of plasma miR-1, cTnI, and CK-MB for AMI was estimated by receiver operating characteristic (ROC) curve analysis. Results There were 127 patients in AMI group, and 107 in non-AMI group, including 82 patients with angina pectoris, 2 with pulmonary embolism, 3 with aortic dissection, 2 with acute pericarditis, 3 with myocarditis, 13 with acute heart failure, and 2 with peptic ulcer. Ninety volunteers were served as healthy controls. There was no difference in clinical characteristics including gender and hyperlipidemia between AMI group and non-AMI group. The expressions of plasma miR-1, cTnI and CK-MB were significantly increased in AMI patients as compared with those of the healthy controls [miR-1 (2-ΔΔCt): 4.32±2.60 vs. 1.44±0.75 and 0.98±0.18, cTnI (μg/L): 3.23 (0.63, 10.70) vs. 0.02 (0.00, 0.17) and 0.00 (0.00, 0.00), CK-MB (U/L): 32.40 (14.20, 95.40) vs. 14.40 (11.20, 17.10) and 8.90 (8.28, 9.50), all P < 0.01]. The expression of plasma miR-1 had a significantly positive correlation with cTnI and CK-MB in AMI patients (r1 = 0.395, r2 = 0.490, both P < 0.000). It was demonstrated by ROC curve analysis that the area under ROC curve (AUC) for the diagnostic value of miR-1 on AMI was 0.905 [95% confidence interval (95%CI) = 0.860-0.950, P = 0.000], the sensitivity was 86.6%, and the specificity was 95.4%; the AUC for cTnI was 0.908 (95%CI = 0.870-0.946, P = 0.000), the sensitivity was 81.9%, and the specificity was 95.9%; the AUC for CK-MB was 0.795 (95%CI = 0.736-0.854, P = 0.000), the sensitivity was 63.0%, and the specificity was 92.9%. Conclusions Plasma miR-1 has the capacity in early diagnosis of AMI, superior to CK-MB, and equal to cTnI. It can provide additional diagnostic information beyond cTnI. The diagnostic accuracy for early AMI can be improved with the combination of plasma miR-1 and cTnI.
ABSTRACT
Objective To explore the sustainable development strategy.of translational medicine in hospitals, we conducted the in-depth analysis for the current status of translational medicine in the Level 3-A hospitals in Wuxi city.Methods the SWOT analysis was performed to identify the advantages, disadvantages, opportunities and threats in the translational medicine development in for those hospitals.Results Development of translational Medicine in those hospitals in Wuxi city highlights the advantages, opportunities and challenges.Conclusions the leadship of Translational Medicine should change their concepts, provide full play to the advantages of a variety of resources, conducting reform and innovation, establish a long-term system of human resource for talented researchers and reward system in order to accelerate the development of translational medicine.
ABSTRACT
Objective To investigate the regulatory effects of miR-146a on inflammation factors production in C ryptococcus neoformans treated THP-1 cells.Methods Cryptcoo ccus neoformans strains were heat killed .Fluorescence quantitative RTP-CR and ELISA were used to detect the levels of IL -1βand TNF-αin THP-1 cells treated with heat-killed Cryptococcus neoformans.In addition, the production of IL-1βand TNF-αwere analyzed before and after Dectin-1or TLR 4 blocked .THP-1 cell lines that were respectively transfected with miR-146 a mimics and inhibitors were constructed and the production of IL-1βand TNF-αin these cell lines were analyzed after interfered with Cryptococcus neoformans.Results With the interference of heat-killed Cryptococcus neoformans, the expression of miR-146a was up-regulated in THP-1 cells, but was down-regulated after Dectin-1 or TLR4 blocked.The expressions of IL-1βand TNF-αinduced by heat-killed Cryptococcus neoformans were enhanced in miR-146a mimics transfected THP-1 cells, but was inhibi-ted in inhibitors transfected THP-1 cells.Conclusion Heat-killed Cryptococcus neoformans could up-regu-late miR-146a expression in THP-1 cells via Dectin-1 and TLR.miR-146a could negatively regulate the ex-pressions of IL-1βand TNF-αinduced by Cryptococcus neoformans.
ABSTRACT
Objective To evaluate the diagnostic performance of plasma miR-499 in acute myocardial infarction (AMI) diagnosis.Methods Diagnostic accuracy test.The suspected AMI patients,who with chest pain for more than half an hour and been admitted in the Second People's Hospital of Wuxi and First People's Hospital of chuzhou during October 2010 and July 2011,were consecutively and prospectively enrolled in the present study.Sixty apparently healthy individuals were designed as healthy control.The plasma samples of the suspected AMI patients were collected within two hours after admission.The plasma miR-499 was determined by real time polymerase chain reaction (RT-PCR).The diagnostic performance of plasma miR-499 for AMI was estimated by receiver operating characteristic (ROC) curve analysis.The association between plasma miR-499 and AMI was analyzed by multivariable logistic model.The plasma miR-499 was determined and explained in blind fashion.Results Two hundred and nine suspected AMI patients,including 131 confirmed AMI patients (46 STEMI and 85 NSTEMI) and 78 AMI free patients were enrolled in the present study.The delta cycle threshold (ΔCT) was 1.01 ± 3.34 for AMI patients,-2.76 ± 2.90 for non-AMI patients and-3.79 ± 2.21 for healthy controls.The differences had statistical significance (x2 =96.77,P ＜ 0.01).The area under curve (AUC) of plasma miR-499 was 0.80 (95％ C I:0.74-0.86),lower than that of cardiac troponin Ⅰ (AUC =0.90,95％ CI:0.86-0.94) on admission (P ＜0.01).At the optimal cut-off of 0.18,the diagnostic sensitivity and specificity were 0.69 (95％ CI:0.61-0.77) and 0.77 (95％ CI:0.66-0.86),respectively.The coefficient of correlation between plasma miR-499 and cTnI was 0.72 (P ＜0.01).The odds ratio (OR) of plasma miR-499 ＞0.18 for AMI was 2.59 (95％ CI:1.10-6.07),after adjusted cTnI.Conclusions Plasma miR-499 is a useful biomarker for AMI diagnosis.It can provide additional diagnostic information beyond cTnI.Combination utility of plasma miR-499 and cTnI may improve the diagnostic accuracy for AMI.
ABSTRACT
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.
ABSTRACT
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.
ABSTRACT
ObjectiveTo investigate mechanisms for IL-27 induced proliferation and differentiation of peripheral blood CD4+ T cells in primary biliary cirrhosis (PBC).MethodsPeriperal blood CD4+ T cells were isolated from patients with PBC,chonic hepatitis B (CHB) and health controls (HCs).After IL-27 stimulation,proliferation ability of CD4+ T cells was evaluated by CCK-8 kit,and cytokines were analyzed by ELISA.Real-time PCR was employed to assay mRNA expression of T-bet and GATA3 in CD4+ T cells.p-STAT-1 and pSTAT-3 expression in CD4+ T cells were detected by Western blot.ResultsEnhanced proliferation of CD4+ T cells was found in all subjects after IL-27 stimulation.However,the proliferation ability in patients with PBC was greater than that in CHB and HCs ( P＜0.001 ).Levels of IL-2 and IFN-γ in supernatant from IL-27-incubated PBC blood CD4+ T cells were higher than that from CHB and HCs (P＜0.001 ).In normal situation,T-bet mRNA of CD4+ T cells in PBC group was higher than that in CHB group (P=0.007).Furthermore,after IL-27 stimulation,elevated T-bet mRNA expression and GATA3 inhibition were found in patients with PBC.High expression of p-STAT-1 and p-STAT-3 in blood CD4+ T cells were found in PBC,CHB and HCs after stimulation by IL-27.But their expression in patients with PBC were higher than those in patients with CHB and HCs.ConclusionProliferation of blood CD4+ T cells could be induced by IL-27 in patients with PBC.The signaling pathways of p-STAT-1,p-STAT-3 were involved to induce Th1 immune response and related cytokines expression.This study implicated that IL-27 may play important roles in early inflammation damage in PBC.
ABSTRACT
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.
ABSTRACT
Objective To explore the role of sialic acid-binding immunoglobulin-like lectin-one (Siglec-1) in the process of atherosclerotic inflammation induced by oxidized low-density hpepmtein (ox-LDL). Methods Ox-LDL was synthesized by oxidization of native LDL and different concentration of ox-LDL was added to the culture medium of RAW264.7. Forty-eight hours later, cells and supernatants were collected separately. The expression of Siglec-1 protein and mRNA were measured by flow cytometry(FCM) and real-time quantitative RT-PCR, respectively. The levels of monocyte chemotactic protein-1 (MCP-1), macrophage inflammatory protein-1α(MIP-1α) and IL-8 in supernatants were determined by ELISA. Re-sults By the stimulation of ox-LDL, Siglec-1 protein and mRNA on RAW264.7 cells were significantly in-creased, meanwhile, the cytokines levels in culture supematants were significantly higher than that in the control group. And both Siglec-1 expression and cytokine secretion were ox-LDL dose-dependent. Conclu-sion Ox-LDL can increase Siglec-1 protein and mRNA expression and some inflammatory cytokines secre-tion on RAW264.7 cells in a dose-dependent manner. Manifested by enhanced Siglec-1 expression, the acti-vated macmphages may take a part in the development and progression of atherosclerosis.
ABSTRACT
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.
ABSTRACT
AIM:Repair of trachea is disturbing the surgeon. Tissue engineering technology will probably resolve this problem. Seed cell is one of the key factors in engineered tracheal cartilage construction. This study investigated the feasibility of constructing tissue-engineered cartilage from porcine bone marrow mesenchymal stem cells(MSCs) cultured and induced in vitro using tissue engineering technique. METHODS:The experiment was performed in the Central Laboratory of Peking Union Medical College Hospital between October 2006 and May 2007. ①By density gradient centrifugation,the MSCs were isolated and purified from porcine bone marrow. The MSCs had been cultured and induced in the defined medium mainly including transforming growth factor-?1,and then the type Ⅱ collagens were detected by immunohistochemical assay. The induced MSCs were seeded onto polyglycolic acids(PGA) scaffold as experimental group,and PGA scaffold were implanted into subcutaneous tissue as control group. The cell-scaffold construct was wrapped around a silicon tube(0.4 cm in diameter) and implanted into subcutaneous tissue of porcine. All specimens were harvested after in vivo culture for 6,8 and 10 weeks and evaluated by gross view,histology,and immunohistochemistry. RESULTS:①The MSCs were obtained by density gradient centrifugation method,and abundant seed cells were obtained after culture and amplification. ②The MSCs differentiated towards chondrocyte when cultured in the specific medium in vitro and were verified by the positive result of collagen type Ⅱ through immunohistochemistry. ③After implanted into subcutaneous tissue for 6,8 and 10 weeks,the cell-scaffold formed a tubular cartilage,which was very similar to normal porcine tracheal cartilage in both gross view and histology. And the result of collagen type Ⅱ through immunohistochemistry was positive. CONCLUSION:The in vivo and vitro cultured MSCs from porcine bone marrow can generate tissue-engineered cartilage under chondrogenic induction.
ABSTRACT
Objective To observe the difference in anticoagulant ability of endothelial cells from different sources.Methods Bone marrow mesenchymal stem cells(BMMSCs)were cultured,purified,and expanded by Ficoll-Paque density gradient centrifugation and adherent culture in vitro.Then they were induced and differentiated in medium with 10 ?g/L VEGF.After 7 days,Von Willebrand factor(VWF)of the cells were identified by immunohistochemistry.At last,the major anticoagulant gene expression of the vascular endothelial-like cells derived from BMMSCs and the human umbilical vein endothelial cells(HUVECs)was detected and compared by reverse transcriptase PCR(RT-PCR).Results Though BMMSCs can successfully differentiate into vascular endothelial-like cells in vitro,they fail to express mRNA of the major anticoagulant gene.However,HUVECs can express the mRNA of these genes.Conclusion BMMSCs can differentiate into vascular endothelial-like cells in vitro,but their anticoagulant ability is inferior to HUVECs.
ABSTRACT
0.05),but was correlated with ?-GT(r=-0.295,P