Objective: This study aimed to investigate the induction of MxA protein in PBMC treated with various doses of adenovirus type 3(Ad3) and to test the antiviral effect of MxA protein against Ad3 in vitro.Methods:The content of MxA protein in cytoplasm of the peripheral blood mononuclear cells( PBMC) , which were treated with various dose of Ad3 was detected by flow cytometry. The antiviral effect of MxA protein against Ad3 in Hela cells was studied by the microdose cytopathogenic effect inhibition assay. Results: MxA protein that in all the cell groups that were treated with various dose of Ad3 was higher than that of control. 10 ng/ml MxA protein can resist 20TCID50 Aad3. Conclusion: It was suggest that MxA protein that can be induced by Ad3 in PBMC and recombinant MxA can resist Ad3.

Background and purpose:Triple-negative breast cancer (TNBC) possesses high risk of relapse and metastasis. Clinically, there are no speciifc targeted-therapies to TNBC except chemotherapy. Therefore, studying the mechanism of relapse and metastasis has signiifcance to improve the patients’ survival rate. This experiment aimed to study the effect of MAPK activation on migration and invasion of triple-negative breast cancer cells. Methods:Difference of migration and invasion between lung-high metastasis breast cancer cell line 231-HM and its parental cell line 231-p were first examined by cell scratch and transwell;Then, metastasis-associated proteins and MAPK-associated molecules were detected by Western blot; Last, 231-p cells were treated with P38/MAPK inhibitor and used to determine cell migration, invasion, and metastasis-associated proteins thereafter. Results:Compared with the parental cell line 231-p, 231-HM cells displayed obviously higher ability of migration and invasion. With the increased expression of Caveolin-1and β-catenin, the phosphorylation of MAPK-associated molecules including P38, Erk1/2, and MEK was highly decreased. Treatment of 231-p cells with low concentration (10 μmol/L) of the P38/MAPK inhibitor SB202190 increased the migration and invasion of 231-p cells, and the expression of Caveolin-1 andβ-catenin. Conclusion:Activation of MAPK signaling inhibits the migration and invasion of triple-negative breast cancer.

Objective To investigate the reliability of extravascular lung water index (EVLWI) and pulmonary vascular permeability index (PVPI) in assessing the severity of acute respiratory distress syndrome (ARDS) in critically ill patients.Methods Forty-six patients with ARDS,who were admitted in our emergency intensive care unit,aged 18-72 yr,weighing 46-72 kg,of Acute Physiology and Chronic Health Evaluation Ⅱ score 11-25,were divided into 3 groups:PaO2/FiO2 ≤ 100 mmHg severe group (n =16);100 mmHg ＜PaO2/FiO2 ≤ 200 mmHg moderate group (n =14);200 mmHg ＜PaO2/FiO2 ≤ 300 mmHg mild group (n=16).Before treatment,and at 24 and 72 h after diagnosis of ARDS,PVPI,EVLWI,cardiac index (CI),and intrathoracic blood volume index (ITBVI) were measured,and blood gas analysis was performed.PaO2/FiO2 was calculated.The 28 day fatality after admission to hospital was recorded.Person correlation of PVPI and EVLWI with PaO2/FiO2,ITBVI and CI was analyzed.Results The PVPI,EVLWI and fatality rate were significantly higher at each time point in moderate group and severe group than in mild group,and in severe group than in moderate group (P＜0.05).The correlation coefficient between PVPI and PaO2/FiO2 was 0.778,and between EVLWI and PaO2/FiO2 was-0.437 (P＜0.05).There was no correlation between CI and ITBVI (P＞0.05).The correlation coefficient between EVLWI and PaO2/FiO2 was-0.448,and between EVLWI and ITBVI was 0.347 (P＜0.05).There was no significant difference between the correlation coefficient between PVPI and PaO2/FiO2 and the correlation coefficient between EVLWI and PaO2/FiO2 (P＜0.05).Conclusion PVPI and EVLWI both can assess the severity of ARDS in critically ill patients,showing a consistent reliability.

Objective To investigate whether MICA gene exon 2,3 and 4 polymorphism is associated with seronegative spondylarthropathies (SpA) or not in Chinese Han population.Methods All 199 B27 positive patients with SpA and 183 randomly ethnically matched healthy controls,and 12 B27 positive controls were enrolled to detect the MICA genotype from its exons 2,3 and 4 by using PCR SSOP method in Shanghai area.Results The MICA007 allele frequency was significantly more in the patient group (18 0%) than in the randomly healthy control group (6 6%) (RR=3 04, P =0 000 045).However,the MICA007 allele frequency was not significantly higher in the B27 positive patient group than in the B27 positive control group.Conclusion The MICA007 allele itself may not be the real disease susceptibility gene involved in the development of ankylosing spondylitis.The increased frequency of MICA007 allele is supposed to be due to a strong linkage disequilibrium between MICA and HLA B genes.

Objective To investigate the effect of the transplantation of autologous marrow mesenchymal stem cells transfected by angiogenin gene in a porcine chronic ischemic beart model.Methods Methods Mesenchymal stem cells were trnsfected byAd/Ang.The pigs underwent placement of amerold occluder around LCx and 4 weeks later sujected to transplantation of the trandected mesenchymal stem cells.The animals were evaluated by coronary angiography,echocardiography,mannetic resonance imaging end pathologic observation.Results All animal showed 95%occlusion fo LCx.4 weeks after treatment ,the perfusion of LCx,left ventricular ejection fracton were greatly evhanced.A large number of labeled mesenchymal stem cells were successfully uncorporated into blood vessels in the ischemic myocardial regions with increased vessels countin and survival of implanted cell.Conclusion Transplantaton of autologous mesenchymal stem cells trendected by angiogenin gene offere obvious advsntases of great improvement of blood supply and the heart funcition.

To better understand the molecular characterization ,pathogenesis ,origin ,and evolutionary relationship of the first human-infecting H6N1 influenza virus ,the dataset was downloaded from the Flu and GISAID databases ,and the phy-logenetic trees were reconstructed by maximum likelihood (ML) ,maximum parsimony (MP) and Bayesian inference (BI) . Furthermore ,the evolution rate ,the most recent common ancestor (TMRCA) ,variable sites ,glycosylation sites ,cleavage sites and other some molecular features were also analyzed by bioinformatic means .The results indicated that the HA gene fell into an individual clade with other three H6N1 virus strains isolated from chicken .TMRCA was in 2002 ,and the cluster had a higher evolution rate .The molecular features revealed the HA protein had 5 N-glycosylation sites ,2 O-glycosylation sites and 2 unique variable sites .The cleavage sites had only one basic amino acid and suggested that the human-infecting H6N1 virus strain was low pathogenic .However ,because of the higher substitution rate and viral reassortment ,people should pay more attention .

Objective To investigate the clinical value of endoscopy for chronic diarrhea in infants and safety of gastroscopy replacing colonoscopy.Methods According to the Nelson chronic diarrhea definition,data of 52 hospitalized infants under 1 year from March 2006 to April 2014,who underwent colonoscopy because of diarrhea lasting for more than 2 weeks and achieving no improvement after series of treatments,or diarrhea suspected with severe milk protein allergy were collected.Endoscopy and mucosal biopsy were performed under intravenous anesthesia.Results A total of 49 cases (94.2%)were found abnormal under endoscopy.Lymphocytes,plasma cells and a little eosinophils were found in lamina propria in 26 cases,20-100 eosinophils/HPF were found in lamina propria in 21 patients,who were diagnosed as eosinophilic colitis.The 21 patients came back for open food challenge test 4 weeks later,16 cases were positive,who were diagnosed as milk protein allergy.Two infants with Crohn disease and ulcerative colitis respectively were treated with mesalazine and prednisone,but symptoms repeated.No complication was observed during endoscopy.Conclusion The etiology of infant chronic diarrhea is complex,except for peptic infections and lactose intolerance ,eosinophilic colitis may be the major cause.Ulcerative colitis and Crohn disease are rare in infants,but cannot be ignored .Endoscopy and mucosal biopsy are important in di-agnosis and treatment.Gastroscopy instead of colonoscopy is safe and effective.

Objective:To investigate influences of two different HLA-B antigens expressed on K562 cells on receptors expression of NK cells from peripheral blood lymphocytes.Methods:Studied the alteration of the percentage of CD16+CD56+ cells and the percentage of KIR3DL1+ cells before and after PBMC interaction with K562 cells for 24 hours,and also compared the percentage of CD16+CD56+ cells and the percentage of KIR3DL1+ cells after PBMC interaction with two different kind of K562 cells transfected with HLA-B39 and HLA-B51 respectively.Results:After PBMCs were incubated with K562 cells for 24 hours,the percentage of CD16+CD56+ cells and the percentage of KIR3DL1+ cells were both increased.However,after PBMCs were incubated with K562-HLA-B51 cells for 24 hours,the percentage of KIR3DL1+ cells and the percentage of CD16+CD56+ cells were both decreased in comparison with that interaction with K562-HLA-B39 cells.Conclusion:CD16 up-regulation was associated with an up-regulation of inhibitory receptors(KIR3DL1).The interaction between HLA-Bw4 and KIR3DL1 would down-regulate the expression of KIR3DL1.In addition,KIR3DL1 down-regulation was associated with down-regulation of activating receptors(CD16).

Objective To investigate optimal extraction process of Piper puberulum ( Benth.) Maxim.and qualitative analyze the chemical component of the extracts.Methods Method of solvent heating reflux was used for extraction.On the basis of single factor experiment, L9 (34 ) orthogonal experiment was designed with the variants of extraction frequency, time, material-liquid ratio, and immersion time.Extraction rate as index, extraction processes were optimized to achieve best extraction.The extracts, including total extract, water elution, and ethanol elution, were physiochemically analysed to achieve an initial qualitative result.Results The optimal extraction process was: extractions 3 times for 2 hours, with an 1︰30 material -liquid ratio and 2 hours of immersion, Initial qualitative analyzed the total extracts containing amino acids, polypeptides, proteins, alkaloids, steroids or triterpenes, flavones, saponins, polysaccharides, reducing sugars or glucosides, cumarins, terpene lactones, phenols, and tannins.The water elution containing: amino acids, polypeptides, proteins, saponins, polysaccharides, reducing sugars or glucosides, cumarins, and terpene lactones.The ethanol elution containing: amino acids, polypeptides, proteins, alkaloids, steroids or triterpenes, flavones, polysaccharides, reducing sugars or glucosides, phenols, and tanins.Conclusion The experiments show that optimal extraction process can achieve high extraction yield, stable and practical.

BACKGROUND:As current studies on isolation, culture andcryopreservation of human amniotic epithelial cells (hAECs) are relatively scattered, it is difficult to form a comprehensive and effective solution to meet the clinical needs of stem cells for transplantation in future.OBJECTIVE:To establish the technology of isolation, culture and cryopreservation of hAECs, and to study the biological characteristics of hAECs.METHODS:Orthogonal method was used to study the effects of different factors on the separation, culture and cryopreservation, and range method was adopted to analyze the data to optimize the separation, culture and cryopreservation. We performed cell primary and passage cultures, morphology observed by microscope, drawn cell growth curve and flow cytometry assay, immunofluorescence staining, hepatocyte like cell differentiation to study the biological characteristics of hAECs.RESULTS AND CONCLUSION:(1) The optimal hAECs separation conditions were as follows:trypsin digestions were conducted at a concentration of 0.25%, four times, once for 20 minutes digestion; optimal conditions of culture were 4×108/L cell seeding density, 10 μg/L epidermal growth factor, 5% serum; optimal conditions of cryopreservation were 1×1010/L cell cryopreservation density, 10% dimethyl sulfoxide, 80% serum. (2) The primary cells were adhered to the wall in 2-3 days, exhibiting irregular polygon, paving stone-like growth. Cell adherence and growth rate were accelerated after subculture, and the growth and proliferation ability of passage 2 cells were not significantly decreased after cryopreservation and resuscitation. (3) Immunofluorescence staining showed that the primary cells strongly expressed SSEA-4 and CK19, but did not express Vimentin, CD45 and HLA-DR. The immunophenotype statistics of the primary and passage 4 cells showed the epithelial mesenchymal transition of hAECs in culture process. (4) Immunofluorescence staining showed that the liver cell marker expression of ALB, CK18 was significantly increased after hAECs were induced to differentiate into hepatocyte-like cells. Glycogen staining revealed glycogen synthesis in hAECs after 3 weeks of induction. To conclude, hAECs are easy to obtain and have strong proliferation ability in vitro, and express surface markers for undifferentiated embryonic stem cells.