0.05) . Four weeks after culture,both transcription factor NKX-2.5 and GATA4 were expressed in the 5-azacytidine group. Additionally,?-mysion heavy chain but not ?-mysion heavy chain expression was observed. CONCLUSION:5-azacytidine induced the differentiation from bone marrow mesenchymal stem cells into myocardial-like cells;in addition,simulated biological microenvironment in both indirect contact group and myocardial cell lysate group also induced the same differentiation. The differentiated cells were cardiac possesses which were between mature cells and cardiac progenitor cells.

Objective To study the changes and clinical significance of glutathione redox-state in patients with chronic heart failure.Methods A total of 81 hospitalized CHF patients(NYHA heart function grade from Ⅱ to Ⅳ)and a total of 28 paroxysmal supraventricular(PSVT)patients without other diseases(controlled group)were recruited in Cadiovascular Wards of the First Affiliated Hospital of Suzhou University from Jun.2006 to Mar.2007.The plasma concentration of GSH and GSSG was determined by using glutathione reductase recycling method(GR-DTNB).Redox potential(EhGSH/GSSG)was calculated using Nernst equation according to the concentration of reduced and oxidized glutathione.Results(1)The concentration of GSH was significantly lower in CHF group compared with control group(P

To study the value of lung ultrasound score (LUS) in assessing the clinical outcome of patients with ventilator-associated pneumonia (VAP).A total of 99 VAP patients were enrolled in a prospective study.All patients met the diagnostic criterion of VAP based on the 2013 guidelines and admitted into our ICU from Jun 2013 to Jun 2015.All parameters were recorded on the diagnostic day (day 1) and day 5,including LUS,clinical pulmonary infection score (CPIS),chest X ray (CXR),Acute Physiology and Chronic Health Evaluation Ⅱ (APACHE Ⅱ) score,Sequential Organ Failure Assessment (SOFA) score,etc.According to the CPIS,patients were divided into 2 groups(CPIS less than 6 and more or equal to 6).CPIS and LUS were similar on day 1 between two groups (P ＞ 0.05).However,on day 5,significant differences of CPIS and LUS were found between groups with CPIS ＜ 6 and CPIS≥6 (P =0.019 and P ＜ 0.001 respectively).LUS decreased on day 5 in CPIS ＜ 6 group and increased in CPIS ≥6 group.In CPIS ＜ 6 group,there was a positive correlation between LUS and CPIS on day 1 (r =0.375,P =0.003) and day 5 (r =0.590,P ＜ 0.001).CPIS ≥6 groupshowed the same trend on day 1 (r =0.484,P =0.002) and day 5 (r =0.407,P =0.011).LUS can be used to dynamically evaluate the clinical outcome of VAP.

Objective To study the clinical and immunologic features of 67 cases of patients with scrub typhus.Methods Epidemiological data,clinical manifestations,laboratory and image examination results and treatment of 67 cases of patients with scrub typhus hospitalized from January 2010 to December 2013 at the First Affiliated Hospital of Nanjing Medical University were retrospectively analyzed.Results Autumn-winter type scrub typhus was predominant in the 67 patents.The main clinical manifestations included hyperpyrexia (100％,67/ 67),eschar or ulcer (91.0％,61/67),rash (52.2％,35/67),lymphadenopathy (46.3％,31/67) and other non-specific symptoms.Laboratory test results:①80.6％ (54/67) of the patients had normal or decreased white blood cell count,62.7％ (42/67) of them with elevated lymphocytes (＞ 40％),thrombocytopenia was detected in 34.3％ (23/67) of the patients,the lowest platelets count was 14 × 109/L.②14 of 17 cases (82.4％) had normal percentage of CD3+ T lymphocytes,CD8+ T lymphocytes increased in 16 of 17 cases (94.1％),while the percentage of CD4+ T lymphocytes (14 of 17,82.4％) and B lymphocytes (15 of 17,88.2％) decreased in most of the patients.③C reactive protein (CRP),ferritin and erythrocyte sedimentation rate (ESR) were elevated in 95.2％ (59/62),93.8％ (45/48) and 89.1％ (49/55) of the patients,respectively.④Elevated alanine aminotransferase (77.6％,52/67) and aspartate aminotransferase (80.6％,54/67) were frequent findings.⑤94.6％ (35/37) of the patients had increased adenosine deaminase.⑥ Antinuclear antibody was positive in 31.0％ (13/42) patients.⑦Epstein-barr virus DNA was detected in 15.6％ (7/ 45) cases,77.6％ (38/49) was accompanied with chest radiographic abnormalities.Two cases had tuberculosis.All the patients recovered after therapy with doxycycline,levofloxacin or moxifloxacin.Conclusion Autumn-winter scrub typhus has typical clinical manifestations in the 67 patients,and it can cause patients' immune disorders.

Objective To compare the differences of cardiac function and interstitial remodeling between diastolic heart failure(DHF) and systolic heart failure(SHF) rabbit models. Methods To establish DHF model with abdo-mial aorta constriction and SHF model with abdomial aorta constriction plus aortic insufficiency. The cardiac func-tion was examined by UCG parameters and homodynamic parameters. The collagen content was measured through hydroxyproline colorimetric assay and shown as collagen area(CA), collagen volume fraction(CVF) and area ratio of Ⅰ to Ⅲ type collagen with PSR. Results Compared with control group, there were significantly increased thick-ness and stiffness of myocardium, impaired diastolic function but normal ejection fraction (EF), and significantly increased collagen content, CA, CVF and area ratio of Ⅰ to Ⅲ type collagen in DHF group; heart chamber was sig-nificantly enlarged, systolic function decreased, and collagen content, CA, CVF significantly increased, but ratio of Ⅰ to Ⅲ type collagen decreased in SHF group(P <0.05 or P <0.01). Conclusion DHF and SHF rabbit mod-els were established successfully, which can simulate clinical profiles and provide technical support to future re-search.

ObjectiveTo evaluate the value of lung ultrasound score (LUS) on assessing the severity and prognosis in patients with acute respiratory distress syndrome (ARDS), and to investigate its correlation with oxygenation index, acute physiology and chronic health evaluationⅡ (APACHEⅡ) score, sequential organ failure assessment (SOFA) score, and clinical pulmonary infection score (CPIS), and other traditional parameters.Methods A prospective double-blind cohort study was conducted. Sixty-two ARDS patients conformed to the Berlin diagnostic criteria admitted to intensive care unit (ICU) of Beijing Huaxin Hospital from October 2013 to December 2014 were enrolled, including 14 cases with mild, 18 moderate, and 30 severe ARDS; among them 37 cases were of ARDS with pulmonary origin, and 25 non-pulmonary ARDS; 35 patients survived, and 27 died. The clinical data and scores of all patients were recorded by one specialized observer, including baseline data, hemodynamic parameters, lactate, respiratory parameters, and APACHEⅡ, SOFA and CPIS scores. Another observer of recording was responsible for the results of lung ultrasound, LUS, and echocardiogram. The correlation between LUS and oxygenation index as well as APACHEⅡ, SOFA and CPIS scores was analyzed by bivariate correlation analysis. Receiver operator characteristic curve (ROC) was plotted, and the predictive value, sensitivity and specificity of mild ARDS, moderate ARDS, severe ARDS and mortality by LUS were calculated. Results LUS had a negative correlation with oxygenation index (r = -0.755,P< 0.001), a good positive correlation with APACHEⅡ (r = 0.504,P< 0.001), SOFA (r = 0.461,P< 0.001) and CPIS (r = 0.571,P< 0.001) was found. LUS in the pulmonary ARDS group had a positive correlation with CPIS (r = 0.399,P< 0.05), and a positive correlation was found in non-pulmonary ARDS group (r = 0.350,P< 0.05), which indicated that the correlation in pulmonary ARDS was more satisfactory than that in non-pulmonary ARDS. LUS in the pulmonary ARDS group was significantly higher than that in non-pulmonary ARDS group (22.1±4.9 vs. 11.3±2.1,t = 11.667,P< 0.001); LUS in mild, moderate, severe ARDS groups was 9.9±1.7, 14.0±1.4, 23.6±4.1. The predictive value for mild ARDS by LUS was 7.0, sensitivity of 87.0%, specificity of 89.0%; that for moderate ARDS was 11.0, sensitivity of 89.0%, specificity of 87.0%; that for severe ARDS was 8.0, sensitivity of 90.0%, specificity of 88.5%. LUS was 24.3±3.8 in the death group, and 12.7±2.9 in the survival group. Area under ROC curve (AUC) was calculated, and the patients with LUS> 19.0 had a high mortality, sensitivity for predicting death was 84.0%, and specificity of 89.0%.Conclusion Bedside LUS, which is simple and easily available, could evaluate the changes in pulmonary ventilation area of ARDS, and its degree of severity, and prognosis including prediction of mortality of the patients.

Objective To analyze human leucocyte antigen (HLA)-A0201 restricted antigen-specific cytotoxic lymphocytes (CTL), and to investigate the difference of T cell response to specific antigen epitopes between patients with acute phase of acute hepatitis B and active phase of chronic hepatitis B. Methods Peripheral blood mononuclear cells (PBMC) from 5 patients with acute phase of acute hepatitis B and 6 patients with active phase of chronic hepatitis B were isolated. The numbers and functions of CD8+ T-lymphocyte epitope peptide specific CTL were detected using enzyme-linked immunosorbent spot (ELISPOT) assay, and the 3 peptides were from HBV polymerase region (Pol575-583), envelope region (Env348-357) and core region (Core18-27), respectively. The data were analyzed using t test. Results The spot formation cell counts (SFC) of Pol575-583, Env348-357 and Core18-27 stimulations in patients with acute phase of acute hepatitis B were 110±13, 165±17 and 185±20, respectively; and those in patients with active phase of chronic hepatitis B were 22±4, 23±5 and 30±5, respectively; the differences were all significant (t=10.9, 15.2 and 8.0, respectively, all P<0.05). The CTL responses to the three peptides in patients with acute phase of acute hepatitis B were Pol575-5830.05). The SFC were increased upon non-antigen specific HLA-A2404 restricted epitope (Core117-125), but the difference was not significant compared with negative control group (P>0.05). Conclusions Hepatitis B virus-specific CTL responses in patients with acute hepatitis B are significantly higher than those in patients with chronic hepatitis B. The number and function of polyclonal CTL are both impaired in patients with chronic hepatitis B.

Aim To observe the effects of simvastatin on PPARγ and p65 subunit of NF-κB and to invest the mechanism of simvastatin preventing hypertrophy and keeping cardiac function.Methods 24 rabbits were divided into 4 groups.Rabbits received sham operation as health control in group I. In other groups, aortic regurgitation and coarctation of ascending aorta were operated in rabbits.Rabbits received no drugs in Group Ⅱ. In group Ⅲ, rabbits were given simvastatin 5 mg·kg~(-1)·d~(-1) after the operation for 8 weeks. In group Ⅳ, rabbits were given simvastatin 5 mg·kg~(-1)·d~(-1) after 4 weeks of operation for 4 weeks. At the beginning and the end of the experiment, left ventricular end diastolic pressure (LVEDP) was measured with catheter. At the end of the experiment, heart weight (HW), left ventricular weight (LVW), body weight (BW), heart weight/body weight radio (HW/BW radio), left ventricular weight/body weight radio (LVW/BW radio) were measured.The PPARγ mRNA expression was analyzed by RT-PCR. PPARγ and p65 protein expression in cardiomyocyte nuclear were analyzed through Western blot. The activity of p65 was analyzed with EMSA.Results The HW, LVW, HW/BW were significantly decreased in the early and late treatment group than in CHF group(P<0.05,P<0.01). The LVW/BW was significantly decreased inearly treatment group than in CHF group, too (P<0.01). The LVEDP was significantly decreased in the early and late treatment group than in CHF group (P<0.01). The mRNA and protein of PPARγ significantly fell in CHF heart (P<0.01). The activity and protein expression of p65 were significantly increased in CHF heart (P<0.01). Simvastatin increased the mRNA and protein expression of PPARγ and decreased the activity and protein expression of p65 (P<0.01).Conclusions Simvastatin inhibits the cardiac hypertrophy and improves cardiac function. The mechanism of simvastatin on cardiac remodeling and function relates to the increase of PPARγ expression and preventing the NF-κB activation.

Objective Clinical evidence has suggested that ATI receptor blocker (ARB) could prevent the development of heart failure. Decreased sareoplasmic reticulum(SR) Ca2+ content, which is due to reduced SR calcium reuptake by SERCA2a, is responsible for defective systolic function in failing heart. To better understand how ARB could improve cardiac systolic dysfunction, we studied the effects of Valsartan on calcium reuptake of SR and its regulatory proteins in heart failure rabbits. Methods Thirty rabbits were divided into three groups: sham rabbits(controls, n= 11), rabbits with heart failure treated with Valsartan (n= 11) and rabbits with heart failure but without Valsartan treatment (n=8).Rabbit heart failure model was established by volume plus pressure overload. Cardiac function was measured by echocardiography. SR calcium uptake was determined by measuring extra vesicular free [Ca2+] changes in a fluores-cence spectrophotometer. SERCA2a, Serl 6-phosphorylated phospholamban (p-PLB), PKA and PP1a protein abundance were deter-mined by use of Western blot analysis. Results Compared to control rabbits, the ejection fractions in the HF rabbits were significantly decreased (P<0.05), these changes could be significantly attenuated by Valsanan treatment (P<0.05).Calcium reuptake of SR, activity of SERCA2a and PKA decreased in heart failing myocytes (P<0.05), with down regulations of p-PLB, SERCA2a and PKA, but up regulation ofPP1αin ventricular samples from the failing rabbits (P<0.05). All of these changes were attenuated by Valsartan treatment (all P<0.05). Conclusion Valsartan improved cardiac function in volume plus pressure overload induced heart failure of rabbits possibly by restoring the SR calcium uptake resulted from attenuating the activities and expressions of SERCA2a and its regulatory proteins.

Objective To observe the relationship of viral load,serum cytokines and tissue damage after severe fever with thrombocytopenia syndrome virus (SFTSV)infection,and to explore the impact of SFTSV levels on tissue injury and prognosis.Methods Twenty-four ambulatory and hospitalized patients who were infected with SFTSV were enrolled between May 2011 and July 2012 at Department of Infectious Disease, First Affiliated Hospital with Nanjiang Medical University. According to their prognosis,they were divided into cure and death group,while 32 healthy blood donators were also enrolled from center blood station in Nanjing as control.The serum SFTSV load was detected using fluorescence quantitative polymerase chain reaction (PCR).The serum T helper (Th)1/Th2/Th17 cytokines in patients with severe fever with thrombocytopenia syndrome (SFTS)were determined dynamically and quantitatively by flow cytometry.The relationships between viral load,cytokines and serum enzymes, white blood cell (WBC),platelet (PLT)counts were analyzed.Comparisons among groups were achieved by rank sum test and correlation analysis among serum cytokines,blood cell counts and tissue damage was done by Spearman correlation test.Results All of the 24 patients showed a positive reaction to SFTSV RNA.The SFTSV loads of 21 cured cases,those of 2 were > 7.0 lg copy/mL,and those of 3 death patients were 6.7 lg copy/mL,8.8 lg copy/mL and 9.8 lg copy/mL,respectively.Serum level of interleukin (IL)-6 (21 .76 pg/mL in day 5 and 7.12 pg/mL in day 7)and IL-10 (14.33 pg/mL in day 5 , 14.13 pg/mL in day 7 and 3.01 pg/mL in day 9)of cured patients were significantly higher than those of healthy controls (IL-6:2.82 pg/mL and IL-10:1 .56 pg/mL)(P <0.05 ).At day 7 and day 9,serum levels of IL-6 of death cases were 137.61 pg/mL and 1 450.83 pg/mL,respectively and serum levels of IL-10 were 50.26 pg/mL and 49.43 pg/mL,respectively.Both of the indicators in the death group were significantly higher than those of cure group (P <0.05 ).However,serum levels of IL-2 and IL-4 were significantly lower than those in healthy control group (P <0.05 ).In the cure group,WBC and PLT counts were lowest during the early course of the disease,and serum alamine aminotransferase (ALT), aspartate aminotransferase (AST ), lactic dehydrogenase (LDH ) and creatine kinase (CK ) were significantly higher than their upper limits of normal.The correlation analysis showed that serum IL-6, IL-10 levels were negatively correlated with PLT count (r=-0.390 and -0.608,respectively;both P <0.01),and positively correlated with SFTSV load (r=0.560 and 0.758,respectively),ALT (r=0.412 and 0.390,respectively),AST (r = 0.686 and 0.764,respectively),LDH (r = 0.633 and 0.677, respectively)and CK (r =0.527 and 0.636,respectively)(all P <0.01 ).Conclusions SFTSV load, IL-6,IL-10 and serum enzyme levels are closely related to the severity of the disease.The inflammatory and anti-inflammatory cytokine storm after SFTSV infection may be involved in the immune pathological injury in patients with SFTS.