Objective To study the specific anti-tumor immunity of dendritic cell(DC) modified by HSP70-tumor peptide complexes and sCD40L in vitro and in vivo.Methods The murine marrow cells were cultured for 7 d,and then randomized to 4 groups: control,only HSP70-H22 peptide complexes,sCD40L,and HSP70-H22 peptide complexes combined with sCD40L.After intervention for 24 h,IL-12 and IL-10 in the supernatant were detected by ELISA assay,CD40 and CD80 of DC by FACS,and the proliferation of spleen lymphocytes by MRL.The ability of spleen lymphocytes activated by DC to H22 cells in vitro was investigated by MTT.The models of murine hepatoma were established,and then randomized to 5 groups(Ⅰ,Ⅱ,Ⅲ,Ⅳ and Ⅴ),followed by interference with normal saline and DC respectively.At 21 d,mice were sacrificed and the weight of tumor was measured.The levels of IL-10 and IFN-? in blood serum were detected by ELISA assay.Results Compared with that in the groups of control,only sCD40L,and HSP70-H22 peptide complexes,the level of IL-10 in the group stimulated by HSP70-H22 peptide complexes combined with sCD40L decreased significantly,but other indexes in this group increased significantly(P

A 69-year-old man with fever,pulmonary nodules,joint pain and superficial lymphadenopathy was admitted to our hospital.The patient had a history of ten year hypertension.She smoked a pack of cigarettes daily for forty years and quitted for fifteen years.Family history of coronary heart disease,diabetes,cancer or other diseases was negative.Chest CT showed a nodule in the left lung lower lobe.Percutaneous lung biopsy revealed a large number of atypical B cell proliferation and infiltration which involved the vessel wall.The atypical B-cell phenotype and genotype was EBERs (+),CD20 (+),CD30 (+),CD15 (-).The patient was diagnosed as pulmonary lymphomatoid granulomatosis (LYG),an angiodestructive and angioinvasive lymphoproliferative disorder which is an Epstein-Barr virus associated B cell disorder with reactive T lymphocytes.The patient received six courses of chemotherapy.In this rare case,misdiagnosis of LYG often occurred due to the complex clinical presentation and non-specific imaging.Percutaneous or open lung biopsy is the main choice in the diagnosis of LYG.

Objective To study the immunomodulatory property of Compound Shougong powder(SGS) on S180 tumor-bearing mice.Methods The S180 model of tumor-bearing mice by Kunming mice were prepared.Then,the tumor-bearing mice were randomized into the model group,positive control group,and SGS groups (29.25g/kg,19.50g/kg,9.75g/kg).Besides the normal group,the other groups were used intragastric administration of into tumor-bearing mice by one time,14 consecutive days.The carbon clearance,quantitative hemolysis and DNFB induced delayed-type hypersensitivity were applied to assay effects of SGS on nonspecific immunity,humoral immunity and cellular immunity.Results Compared with the model control group,in the SGS groups,the tumor-bearing mice with spleen and thymus index of organ improved (all P ＜ 0.05),and the clearance index and values of phagocytic index were elevated(all P ＜0.05),and the productions of IgM and IgG in serum and hemolysin in splenocytes were enhanced(all P ＜0.05),and the percentages of T cells expressing CD4+,CD8+ and the ratio of two subset of T lymphocyte increased,and the IL-2 production of spleen lymphocytes was also improved (all P ＜ 0.05).Conclusion SGS showed significant immunomodulatory property on tumor-bearing mice through specific and nonspecific immunity.

Objective To observe the phosphorylation levels of extracellular signal-regulated kinase 5 (ERK5) in acute myocardial infarction (AMI) patients and the effects of ERK5 selective inhibitor XMD8-92 on human platelet activation in vitro,and to explore its mechanism.Methods Western blot was applied to detect the phosphorylation levels of ERK5,Akt473 and Akt308 in AMI patients (n =34) and stable angina patients (n =33,control).The effects of different concentration of XMD8-92 on human platelet aggregation induced by collagen was tested by aggregometer in vitro.The release of ATP was measured simultaneously by luciferase detection.The effects of XMD8-92 on integrin aIIbβ3 were detected by platelet spreading on immobilized fibrinogen and clot retraction.The effects of XMD8-92 on phosphorylation levels of Akt473,Akt308 PTEN370 and ERK5 were detected by Western blot.Results The levels of phosphor-Akt473,Akt308 and phosphor-ERK5 were significantly higher in AMI patients than that in control group (P＜0.05).ERK5 inhibitor XMD8-92 diminished collagen-induced platelet aggregation,ATP secretion,the average area of platelet spreading on immobilized fibrinogen and the clot retraction extent.The levels of phosphor-Akt (Ser-473/Thr-308) and phosphor-PTEN (Ser370) were significantly down-regulated in the presence of XMD8-92.Conclusions ERK5 plays a role in platelet activation in AMI process.It regulates platelet activation by regulating PTEN and Akt phosphorylation.Its specific inhibitor is hoped to be new antithrombotic drug.

Objective Based on dual channel melting curve analysis-based assay,we developed a method to rapidly detect the drug-resistant mutations in Mycobacterium tuberculosis through real-time PCR.Methods According to the common first-line drug-resistant mutations of Mycobacterium tuberculosis,we designed six dual-labeled fluorescence probes to rapidly detect the drug-resistant mutations through realtime PCR melting curve after amplifications of drug-resistant related gene region of DNA.The targets include rpoB 81 bp core region,katG315,inhA promoter,ahpC promoter and embB306.To validate the sensitivity and specificity of our method,we performed real-time PCR assays to detect drug-resistant mutations in 76 clinical MDR-TB samples,which were collected by Shanghai CDC in 2008.Results In the validation,this method successfully detected drug-resistant mutations in all 76 clinical MDR-TB samples.The △Tm of mutations were from 1.8 to 14.4 ℃.Comparing with the sequencing data,all mutations covered by the six probes were detected with 100％ sensitivity and 100％ specificity (rpoB,80/80; inhA,7/7 ; katG315,59/ 59;ahpC,8/8;embB306,27/27).This method can successfully detect drug-resistant mutations from 100 copies/μl DNA samples.Conclusions A widely applicable real-time PCR assay to detect first line drug-resistant mutations of Mycobacterium tuberculosis has benn developed.This method has proven to have the advantages of high sensitivity,specificity and low risk of contamination.It can be used in rapid diagnosis of clinical drug-resistant tuberculosis and the evaluation of laboratory drug sensitivity test.

Objective To investigate the activation of nuclear factor-B (NF-B)and hypoxia-inducible factor-1 (HIF-1)in patients with obstructive sleep apnea-hypopnea syndrome (OSAHS). Methods Subjects were classified into mild-to-moderate OSHAS group (n = 16), severe OSAHS group (n = 14) and the matched control group (n=30). Gene and protein expressions of NF-B and HIF-1 were measured by RT-PCR, Western blot in peripheral blood mononuclear cells (PBMC). Results NF-κB p65 mRNA and NF-κB p65 protein,HIF-1α mRNA and HIF-1α protein in PBMC were significantly higher in severe OSAHS group than those in mild–to-moderate group and control group (P<0.01). But there were no significant difference in NF-κB p65 mRNA and NF-κB p65 protein between mild-to-moderate group and control group (P=0.068, P=0.254 respectively). Only gene (P<0.05) not the protein (P=0.777) of HIF-1αwas higher in mild-to-moderate as compared with control group. Both NF-κB p65 mRNA and HIF-1αmRNA were positively correlated with AHI (r=0.493, P=0.006, r=0.508, P=0.004), while negatively correlated with nighttime lowest blood oxygen saturation (LSaO2)(r=-0.488, P=0.006, r=-0.46, P=0.011). There was a positive correlation between NF-κB p65 protein level and AHI (r=0.669, P<0.001). HIF-1αprotein level was positively correlated with AHI, ODI (r=0.628, P=0.001;r=0.480, P=0.018). There were positive correlations between NF-κBp65mRNA and HIF-1αmRNA (r=0.543, P=0.002), NF-κBp65 and HIF-1αprotein respectively (r=0.716, P<0.001). Conclusions As the gene and protein of NF-κB and HIF-1 were up-regulated in patients with OSAHS, and also positively correlated with the severity of sickness. We conclude that both NF-κB and HIF-1 were involed in the pathogenesis of OSAHS.

OBJECTIVED: To explore the prevalence and risk factors of type 2 diabetes mellitus (T2DM) and impaired glucose regulation (IGR) in Zhangzi Island of Dalian, additionally to provide a better understanding of further intervention. METHODS: A cross-sectional study was performed in 907 residents aged over 18 years from Zhangzi Island of Dalian. The questionnaire, physical measurement, food intake frequency and previously dietary investigation were used in the analysis. Simultaneously, a series of biochemistrical and immune index was measured, including blood uric acid, blood sugar, total cholesterol, triacylglycerol, as well as serum insulin.RESULTS: The recovery rate of questionnaire was 95.9%, which demonstrated that the prevalence rates of T2DM and IGR in Zhangzi Island were 11.36% and 37.38%, there was no significant differences between males and females (P > 0.05). Only 29.13% of the people with T2DM were aware of the condition. The prevalence of DM and IGR were increased significantly with age (P < 0.01). The results of the logistic regression showed that age, education, alcohol drinking and hypercholesteremia were the risk factors of IGR. In addition, age, education, hypercholesteremia, hypertriglyceridemia, body mass index and hypertension were the important risk factors for the development of DM. CONCLUSION: The prevalence rates of T2DM and IGR in Zhangzi Island of Dalian are high with a low awareness. Health promotion on lifestyle changes may help to increase the public awareness and reduce the prevalence.

Objective To explore the relationship between X - linked spondyloepiphyseal dysplasia tarda (SEDL) gene escaping X chromosome inactivation( XCI) and SEDL phenotype. Methods RT - PCR was performed on total RNA which was isolated from blood samples of patients, female carriers and controls. Patients and female carriers were selected from the pedigree with SEDL caused by the mutation (IVS2 - 2A→C) of the gene. cDNA was analyzed by polyacrylamide gelelectrophoresis(PAGE). Results PAGE data indicateed that female carriers expressed both normal and mutant SEDL mRNA,meaning the SEDL gene escaping XCI. Family investigation showed carrier females in the SEDL pedigree presented no symptoms. Conclusions The SEDL gene escaping X chromosome in-activation is firstly identified from human body. This may explain that carrier females present no symptoms.

Objective:To investigate the expression of Survivin and it's relationship with the expression of Bcl-2 in sinonasal inverted papilloma (SNIP).Method:Immunohistochemical method was used to determine the expression of Survivin and Bcl-2 in 30 cases of SNIP,10 cases of sinonasal squamous cell carcinoma(SCC) and 10 cases of normal inferior concha tissues.Result:Survivin was expressed in 22 of 30(73.3%)cases of SNIP,8 of 10(80.0%)cases of SCC and not expressed in 10(0%)cases of normal inferior concha tissues.Expression of Survivin was significantly higher in SNIP and SCC than in normal tissues.Bcl-2 was expressed in 9 of 10(90.0%)cases of SCC and 2 of 10(20.0%)cases of normal inferior concha tissues.Expression of Bcl-2 was significantly higher in SCC than in normal tissues.Bcl-2 was expressed in 14 of 30(46.7%)cases of SNIP,higher than normal tissues.Expression of Bcl-2 was positively related to expression of Survivin.Conclusion:Survivin may play an important role in the pathway of progression of SNIP and SCC.It may be identified as a new therapeutic target.Bcl-2 may play a synergic role with Survivin in progression of SNIP.

Objective To analyze the pathologically confirmed pulmonary Actinomycosis in the 11 patients in focusing on clinical features and mis-diagnostic reasons so as to improve physicians' awareness of this rare disease and reduce the misdiagnosis.Methods We retrospectively reviewed the medical records of 11 cases with pathologically confirmed pulmonary Actinomycosis during January 2003-August 2015.The clinical data and main causes of misdiagnosis in these cases were collected and analyzed.Results The study included 11 patients with a mean age of(53.0 ± 11.6.0)years.Among the 11 cases,8 (72.7 ％) patients had complications,6 (54.5 ％) were current or ex-smokers.Main clinical manifestations of 11 cases were cough(11/11,100.0 ％),sputum(11/11,100.0 ％),hemoptysis (7/11,63.6％),chest pain(6/11,54.5％)and fever(3/11,27.3％).Ten patients presented with one lobe of lung lesions,including 4 patients in the lower lobe and 3 in the upper lobe of the left lung,2 in the upper lobe and 1 in the lower lobe of the right lung.While,the remained one case presented with lesion locating in right main bronchus.Iconography often presented as pulmonary mass shadow,consolidation shadow,spicule sign,lobulation sign,hilar and/or mediastinal lymphadenopathy and pleural effusion.Vacuolar lesions were observed in some of the focuses.Flexible bronchoscopy was performed in 8 (72.7％)patients.Among them,7 patients showed mucosal swelling and congestion,luminal occlusion with purulence secretion,2 cases with polypoid neoplasm.Initial misdiagnosis rate were 100％ (11/11),among which 7 cases were misdiagnosed as lung cancer,2 cases as fungus infection,and 1 case as pulmonary tuberculosis and 1 case as pneumonia,respectively.All patients were definitely diagnosed by biopsy finding an evidence of hyphae of Actinomycosis in lung tissue specimens.The definitive diagnosis was made by CT-guided percutaneous lung biopsy in 4 cases,by transbronchial lung biopsy (TBLB)in 5 cases and by thoracotomy or video-assisted thoracoscopic surgery(VATS) in 1 case respectively.Actinomycosis in most patients was cured with high-dose penicillin administration over a prolonged period.Conclusions The diagnosis of pulmonary Actinomycosis remains challenging via its non-specific clinical symptoms and iconography features,and the presence of comorbidity may further increase the difficulty and complexity of diagnosis,leading to delaying-or mistaking-diagnosis.Obtaining positively pathological specimens is diagnostic key.Transbronchial lung biopsy through a bronchoscope and CT-guided percutaneous needle biopsy are the priority methods.