Objective To observe the effects of cigarette smoke extract (CSE) on the proliferation and secretion of hydrogen peroxide (H2O2) in human lung fibroblasts (HLFs) induced by transforming growth faetor-β1(TGF-β1).Methods Cultured HLFs were divided into a normal group and a model group induced by TGF-β1 (5 ng/mL),then intervened with CSE at different concentrations (0% ,2.5% ,5%, 10%, respectively).Brdu ELISA assay was used to detect cell proliferation.H2O2release from cultured cells was assayed using a fluorimetric method.Cellular localization of H2O2 and expression of α-SMA were performed using a fluorescent-labeling strategy.Results TGF-β1 stimulated group showed positive expression of α-SMA, implying TGF-β1 had induced fibroblasts to differentiate into myofibroblasts.In TGF-β1 stimulated group,2.5% and 5% CSE promoted cell proliferation (P < 0.01 or 0.05), while 10% CSE inhibited cell proliferation (P < 0.01).In the normal group, both low and high concentration of CSE inhibited cell proliferation (P < 0.01 or P < 0.05), and the inhibition effect was dose-dependent.HLF induced by TGF-β1 generated low constitutive levels of extracellular H2O2 that was markedly enhanced by CSE treatment (P < 0.01).Pretreatment with DPI, an inhibitor of NADPH oxidase, abolished secretion of H2O2.Cellular localization of H2O2 by a fluorescent-labeling strategy demonstrated that extracellular secretion of H2O2 is specific to the myofibroblast.Conclusions Low concentration of CSE can promote myofibroblast proliferation, and markedly increase extracellular secretion of H2O2.CSE possibly take part in the development and progress of idiopathic pulmonary fibrosis by increasing oxidative stress.

Objective To observe the clinical features of abdominal aortic aneurysm(AAA)in the elderly. Methods Seventy-eight elderly cases with AAA were studied retrospectively,the effect of age and AAA size on the growth of AAA were analyzed. Results Risk factors such as smoking,hypertension,hypercholesteremia and artherosclerosis were found in most patients. Aneurysm in other side was found in 25.6%of the patients.Rupture occurred in patients with a larger in size or rapidly developing AAA.The average AAA diameter inerease was 0.1 4 cm/year and it was similar for each age grade.When the AAA diameter was wider than 5 cm,it developed faster.and the growth velocity increased to 0.46 cm/year. Conclusions Age is not a risk factor that affects the development of AAA.If the AAA diameter is≥5 cm,the AAA develops quickly,then active interventions are recommended.

Objective To analyze the clinical characteristics of infective endoearditis complicated with intracranial hemorrhage.Methods The clinical characteristics,brain CT and MRI results and prognosis of 7 patients with infective endocarditis and intracranial hemorrhage admitted between January 1991 and May 2015 were retrospectively analyzed.Results All seven patients presented varying degrees of fever,cardiac murmur and neurological symptoms.The continuous fever lasted from 21 d to 227 d (average 91 d).Mitral valve (4 cases) and aortic valve (3 cases) were the most common affected ones.The brain CT or MRI scan presented intracranial hemorrhage in 6 cases.More than 2 hemorrhage locations were found in 5 patients and parietal lobe was most common one (4 cases).All patients were treated with antimicrobial agents and three patients underwent valve replacement surgery.Three patients were cured and another 2 died due to fatal hemorrhage.Conclusions Patients with long fever history and ineffective antimicrobial treatment are prone to intracranial hemorrhage.Brain CT or MRI scan need be performed in infective endocardits when neurological symptoms are present,and brain angiography should be also considered if necessary.

Objective To evaluate the prognostic values of leukocyte count,hemoglobin,biochemical parameters,erythrocyte sedimentation rate and immunoglobulin on mortality in patients aged 80 years and over.Methods Totally 342 patients(aged 85.6±4.0 years)were followed up for (82.0±36.9) months,and the cause and time of death were recorded.Results During the period of follow up,198 patients suffered from death.Compared with the survival group (132 cases),the death group had older age [ (86.5±4.4)years vs.(84.5±3.2)years,t=-4.86,P＜0.01 ],higher white blood cell [ (6.2± 1.7) ＞ 109/L vs.(5.5±1.3) × 109/L,t=-3.93,P＜0.01 ],lower hemoglobin [(134.4±14.4)g/L vs.(140.0± 12.6)g/L,t= 3.65,P＜0.01 ],slightly faster erythrocyte sedimentation rate [ 11 mm/h(15 mm/h) vs.9 mm/h (10 mm/h),U=- 3.31,P＜0.01 ],lower immunoglobulin M [ (0.9±0.5)mg/L vs.(1.1±0.8)mg/L,t =2.55,P＜0.05 ],slightly higher urea nitrogen [ (7.5±2.6) mmol/L vs.(6.8±1.6) mmol/L,t=2.81,P＜0.01]and creatinine [(113.0±32.5) μmol/L vs.(100.5±15.8) μmol/L,t=-4.65,P＜0.01 ].Cox multivariate analysis revealed that older age (RR=1.083,95％CI:1.040 1.127,P＜0.01),white blood cell count (RR=1.134,95％CI:1.021-1.260,P＜0.05),creatinine (RR=1.011,95％CI=1.0021.020,P＜0.05),hemoglobin(RR=0.835,95％CI:0.714-0.975,P＜0.05)andimmunoglobulin M(RR=0.710,95％CI:0.521-0.966,P＜0.03),aorticaneurysm(RR=2.144,95％CI:1.163-3.951,P ＜ 0.05 ) were the independent risk factors for death.Conclusions Aging,increased WBC count,decreased hemoglobin and immunoglobulin M,elevated creatinine and aortic aneurysm are the independent risk factors for death,which are powerful parameters for the prognostic evaluation in the elderly aged 80 years and over.

Objective To observe the relationship of the red blood cell distribution width and white blood cell count with coronary heart disease and coronary artery lesions.Methods Totally 590 patients undergoing coronary angiography were selected and divided into two groups based on the results of coronary angiography:coronary heart disease group (n=383) and control group (n=207).Based on the number of coronary lesions,patients in coronary heart disease were divided into different subgroup.The Gensini scores of coronary lesions were assessed.The differences in red blood cell distribution and white blood cell count were compared among different groups,and the correlations of coronary lesions with red blood cell distribution and white blood cell count were analyzed.Results The red blood cell distribution and white blood cell count were higher in coronary heart disease group than in control group [(13.06±0.57)％ vs.(12.63±0.49)％,(6.33±1.56) 109/L vs.(5.86± 1.29) 109/L,t=9.771 and 3.728,both P=0.000].The red blood cell distribution and white blood cell count were increased along with the increasing number of coronary lesions (F=51.454 and 7.544,both P=0.000),and positively correlated with the Gensini score (r=0.414 and 0.111,P =0.000 and 0.030).The red blood cell distribution was positively correlated with white blood cell count (r=0.108,P=0.009).The receiver operating characteristic(ROC) curve for red blood cell distribution predicting coronary heart disease showed that the threshold value of red blood cell distribution was 12.75％ and the area under the ROC curve was 0.723 (95％ Cl:0.680-0.765) with a sensitivity of 67.6％ and specificity of 65.2％ for the diagnosis of coronary heart disease.Conclusions Red blood cell distribution and white blood cell count are significantly increased in patients with coronary heart disease and are independently correlated with the severity of coronary lesions.Red blood cell distribution and white blood cell count are independent predictiors for coronary artery disease.

Objective To observe the effect of nitroglycerin(NTG) on pulse wave velocity(PWV) in patients with hypertension(HTN).Methods36 volunteers,mean age 48.1±10.2 years,were divided into HTN group and non-hypertension(NHTN) group according to whether he or she had hypertension or not.The baseline PWV and PWV at 5th minute and 10th minute after sublingual NTG were detected.PWVs of the two groups were compared.ResultsThe basal PWV in HTN group was higher than that in NHTN group.PWV was reduced significantly after NTG were given sublingually 5 minutes or 10 minutes compared with baseline condition(both P<0.01).PWV 10 minutes after sublingual NTG raised a little compared with that after 5 minutes(P<0.01).PWVs 5 minutes or 10 minutes after sublingual NTG in HTN group were higher than those in NHTN group(both P<0.05).ConclusionNTG can reduce PWV in patients with HTN.

Objective To explore in vitro cytotoxic activities of DCIKs against hepatocarcinoma cells by co-culturing cytokine-induced killer cells (CIKs) with dendritic cells (DCs) derived from peripheral blood of patients with hepatocellular carcinoma (HCC).Methods Peripheral blood mononuclear cells (PBMCs) were isolated from 23 patients with HCC and cultured with cytokines to induce DCs and CIKs.DCIKs were induced by co-culturing CIKs with DCs.After 14 days of co-culture,the phenotypes of DCIKs and CIKs were analyzed by flow cytometry,and their in vitro cytotoxic activities against SMCC-7721 and HepG2 hepatocarcinoma cells were measured by MTT assay.Levels of IL-12,IL-4 and IFN-γ in the supernatants of cell culture were detected by enzyme-linked immunosorbent assay (ELISA).Results High expressions of CD3+CD8+ and CD3+CD56+ were observed on DCIKs.The percentages of effector cells,cytotoxic activity and cytokine secretion were all significantly increased with DCIKs as compared with those CIKs without DC co-culture (P＜0.05).Conclusion Co-culture of CIKs with DCs can enhance the differentiation of effector cells and the cytolytic activities of CIKs against hepatocarcinoma cells in vitro.Immunotherapy with DCIKs may be a promising strategy for the treatment of patients with HCC.

BACKGROUND: Nerve growth factor is secreted and synthetized by a variety of cells, such as inflammatory calls and repairing calls, its biological effects are diverse and closely related to the process of wound repair, but its mechanism is not yet clear.OBJECTIVE: To observe the influence of nerve growth factor on the biological characteristics of scar fibroblasts.METHODS: Eight clinical surgical resection specimens, including 5 face and neck hyperplastic scar or keloid specimens, did not receive any treatment; three were prepuce specimens following circumcision (normal tissue). By use of tissue block method, the scar and normal skin fibroblasts were cultured, followed by digestion passage. The scar tissue and normal tissue flbroblasts at 3-6passages in the logarithmic phase were seeded in 96-well plate and divided into the experimental group (scar flbroblest group) and the control group (normal skin fibroblasts group), with two parallel holes in each group were added with 3,33, 0.33 mg/L nerve growth factor, 50 μL. Inverted microscope was used to observe fibroblast morphology. At 24, 48, 72 hours after culture, the absorbanca value was measured using MTT. Fibroblast DNA content and cell apoptosis were determined by flow cytometry.RESULTS AND CONCLUSION: The fibroblasts were adherent cells, the scar and normal skin tissues were shown to cell free out of tissue block and gradual expansion at 4-6 days after incubation. Compared with normal skin fibroblasts, the pathological scar fibroblasts became larger, irregular shape and arrangement. MTT results showed that nerve growth factor could promote the normal and hypertrophic scar fibroblasts growth, which becomes more apparent. Flow cytometry results showed that by adding nerve growth factor, the percentage of scar fibroblasts at proliferating S-G_2-M phase was higher than that in the control;group; with a Iower level of apoptosis. It is indicated that nerve growth factor plays an obviously promoting role on normal and scar skin fibroblasts growth and proliferation, especially on the scar skin.

In vitro cell culture of head and neck tumors and chemosensitivity test to select sensitivity antitumor bogs for individual chemotherapy. Methods:The chemosensitivity of 32 fresh specimens of human head and neck tumors were tested with MTT assay and eight atitumor drugs． Results:The sensitivity was 84.4% for 5-Fu, 2l.9% for VCR, 62.5% for MTX, 8l.3% for BLM, 8l.3% for DDP, 40.6% for ADM, 65.6% for PYM and 3.l% for CTX． The results of chemotherapy with sensitivity drugs of 32 patients were encouraged． There were no serious side-effects in the chemotherapy and no recurrence and metastasis after the treatment． 32 patients were still alive． Conclusion: Chemosensitivity test of tumor cell lines with MTT assay was accuracy， fast，simple，ecoromical，effective method and was valuable for chemotherapy．

Objective To explore the mechanism of cytokines for the scars,and to study the effect of platelet derived growth factor(PDGF)on the biological behavior of fibroblasts in scars.Methods Fibroblasts of scars and normal skins were cultured in vitro.The results were observed and analyzed by light inverted microscopy(LM),and 3-(4,5-dimethyl-thiazol-2-yl)-2,5 ciphenyl tetrazolium bromide (MTT)assay.The effects of PDGF on the biological behaviors of fibroblasts of scars were also determined. Results In vitro study,using LM,FCM and MTT assay,showed that proliferation of fibroblasts were inereased significantly when PDGF was added to the cultures,as compared to the control groups.Conclusions PDGF can increase fibroblast proliferation.These results demonstrate that PDGF is beneficial for wound healing at early stage.