Virus infection can cause a variety of host immune response to prevent the spreading of virus.Innate immunity is the first line of defense to resist to virus infection in host.After the virus enters,the macrophage phagocytoses and process of virus antigen.On the other hand,it activates the innate antiviral immune response.The expression of interferon after EV71 infection may be associated with cell specific response.Most studies have shown that EV71 can inhibit the expression of interferon to achieve immune escape.Then antigen presenting cells present antigens to T/B lymphocytes,once specific T/B lymphocytes are activated by antigen,they will be activated,proliferated,diferentiated to immunologic effector cells,which secrete antibodies,cytokines,cell toxicity of medium,ultimately,develop immunological effect.Both humoral immunity and cellular immunity are activated in EV71 infection,and play an important role in mediating protection effect.Cellular immunity may be associated with the clinical outcome of EV71 infection.The EV71-specific neutralizing antibody generated by the humoral immunity plays a key role in protective immunity,but the antibody response level is not correlated with disease severity.In addition,virus infection can induce cell apoptosis to prevent the virus proliferation and the spreading of progeny virus.

Objective To explore the effects of adipose tissue-derived stem cells (ADSCs) on the proliferation and invasion of pancreatic cancer cell lines in vitro.Methods ADSCs were isolated and co-cultured with pancreatic cancer cells.The proliferation and invasion of pancreatic cancer cells was tested by CCK-8 test kit.Stromal cell derived factor (SDF-1),vascular endothelial growth factor(VEGF),matrix metalloproteinase(MMP-9)and transforming growth factor-β (TGF-β1)in the culture medium were assayed by ELISA.The expression of cytokines in pancreatic cancer cells and ADSCs were detected by qRT-PCR.CCK-8 test kit was used to measure the AMD3100 regulation for the co-culture of ADSCs and pancreatic cancer cells.Results ADSCs can promote the proliferation and invasion of pancreatic cancer cells (all P ＜0.05); more SDF-1,VEGF,MMP-9 and less TGF-β1 was secreted by ADSCs than by pancreatic cancer cells(SDF-1:1100±100 vs.0 vs.0,F=389.134,P＜0.01;VEGF:140±4 vs.99±5 vs.93 ±4,F=174.102,P ＜0.05;MMP-9:61.8 ±4.2 vs.43.5 ±2.8 vs.54.5 ±3.0,F=76.279,P＜0.05;TGF-β1:20.6 ±3.0 vs.35.6 ±2.6 vs.41.3 ±5.5,F =79.338,P ＜0.05).ADSCs promote the expression of VEGF and MMP-9 in pancreatic cancer cells(VEGF:63.7 ±5.9 vs.50.6 ±4.1,t =7.536,P＜0.05; MMP-9:mRNA(55.8±3.6 vs.42.7 ±3.1,t =8.279,P＜0.05).AMD3100 significantly downregulates these growth-promoting effects of ADSCs on pancreatic cancer cells(SW1990:1.539 ±0.140 vs.1.361±0.066,t=2.835,P＜0.05;PANC-1:1.376±0.100 vs.1.281±0.031,t=2.860,P＜0.05).Conclusions ADSCs promote the proliferation and invasion of pancreatic cancer cells.

Objective To compare the difference between bone marrow stomal cell (BMSC) and adipose-derived stem cell (ADSC) of liver fibrosis in rats.Methods BMSC and ADSC of Sprague-Dawley (SD) rats were isolated and purified.The stem cell markers were detected with flow cytometry.The coculture system was set up with 0.4 μm Transwell insert semipermeable membrane.ADSC or BMSC were co-cultured with hepatic stellate cells (HSC).Normal hepatocyte cell line of rat (BRL) was co-cultured with HSC as negative control group and HSC cultured alone was blank control group.After cultured for 72 hours,the proliferation of HSC was determined by cell counting kit-8 (CCK-8) method.The expression of α-smooth muscle actin (α-SMA) of HSC was detected by Western blotting.The apoptosis of HSC was examined by flow cytometry.After BMSC,ADSC and BRL cultured alone for 72 hours,expression level vascular endothelial growth factor (VEGF),interleukin-10 (IL-10),nerve growth factor (NGF) and transforming growth factor-β1 (TGF-β1) in the culture medium were detected by enzymelinked immunosorbent assay (ELISA) method.The rats model of liver fibrosis were established.The rats were divided into BMSC treatment group,ADSC treatment group,BRL group and culture medium group,six rats in each group,which were injected with 1.5 mL BMSC,ADSC and BRL cells suspension (5 × 106) through portal vein,respectively,and same volume of culture medium was injected to the rate of culture medium group,once every two weeks for four weeks.The pathological changes of liver tissue sections were observed and liver fibrosis markers were tested.T test was performed for comparison between two samples and analysis of variance was used for comparison among multiple groups.Results BMSC and ADSC were successfully isolated and cultured.The phenotype of BMSC and ADSC was similar.Compared with blank control group and negative control group,both ADSC and BMSC could inhibit the proliferation of HSC and promote apoptosis (proliferation,2.43±0.27,2.39±0.33,1.92±0.38 and 2.18±0.31,FBMSC =25.61,FADSC =38.63,both P＜0.05 ;apoptosis rate,(5.59 ± 0.40)％,(6.82±0.57)％,(8.31± 1.03) ％ and (9.36 ± 0.54) ％,FBMSC =73.69,FADSC =97.41,both P＜ 0.05).The effects of ADSC were more significant than those of BMSC (t=5.76 and 5.18,both P＜0.05).There was difference in the cytokine levels secreted by ADSC and BMSC (NGF,(7.46 ± 0.54) pg/mL vs (3.95 ± 0.71) pg/mL,t =10.92,P＜0.05; TGF-β1,(8.79 ±0.93) pg/mL vs (6.36±0.85) pg/mL,t=7.58,P＜0.05).The cell transplantation experiment indicated that both BMSC and ADSC had significant inhibitory effect on liver fibrosis.The activity index of inflammation and degree of fibrosis in BMSC treatment group and ADSCs treatment group were 9.87±2.07,4.17 ± 0.94 and 10.13 ± 1.81,3.98 ± 0.82,which were significantly lower than those in blank control group (13.78±2.53 and 5.09±1.15)and negative control group (13.34± 1.89 and 4.95± 1.22,FBMSC=51.26 and 32.29,P＜0.05; FADSC =46.73 and 40.94,P＜0.05).The level of hyaluronic acid ((191.5±33.2) μg/L and (178.8±28.2) μg/L),type Ⅲ collagen ((19.9±5.1) μg/L and (21.7± 3.3) μg/L) and hydroxyproline ((312.6±38.8) μg/g and (325.8±28.2) μg/g) of BMSC treatment group and ADSC treatment group were significantly lower than those of negative control group and blank control group (hyaluronic acid,(282.3 ± 18.7) μg/L and (287.5 ± 26.7) μg/L),F =73.51 ; type Ⅲ collagen,(35.3± 3.3) μg/L and (32.5±4.3) μg/L,F=76.19; hydroxyproline,(458.4 ± 38.1) μg/g and (473.9 ± 63.7) μg/g,F=60.37,all P＜0.05).However,there was no difference between BMSC treatment group and ADSC treatment (all P＜0.05).Conclusions ADSC and BMSC had similar stem cell characteristics.There was difference in inhibiting the activation of HSC between ADSC and BMSC.But there was no significant difference in inhibiting liver fibrosis of rats in vivo.

Objective To explore the relationship between angiogenin-1/2 (Ang-1/2) and clinical parameters of idiopathic pulmonary fibrosis (IPF), and to assess the value of Ang-1/2 in predicting the prognosis of patients with IPF.Methods A retrospective analysis was conducted. Ninety-one patients diagnosed as IPF by high resolution CT (HRCT) and lung biopsy admitted to Daqing Oil Field General Hospitalfrom March 2014 to January 2015 were enrolled. The general data, serum parameters and pulmonary function parameters of all patients were collected. After treatment, all of the 91 patients were followed-up to 2 years. The patients were divided into favorable prognosis group and unfavorable prognosis group according to follow-up results. The differences in all parameters between the two groups werecompared. The relationship between Ang-1, Ang-2 and lung function parameters was analyzed by Pearson correlation analysis. Cox proportional hazard regression model was used to evaluate the effect of clinical parameters on the prognosis of patients with IPF. The effect of Ang-2 in predicting prognosis of patients with IPF was analyzed by receiver operating characteristic (ROC) curve.Results During the 2-year follow-up period, 30 of 91 patients showed a favorable prognosis, and 55 showed an unfavorable prognosis with a poor prognosis rate of 64.71%, and 6 patients withdrew from the study due to loss of follow-up and death. Compared with the favorable prognosis group, Ang-2 level in the unfavorable prognosis group was significantly increased (μg/L: 2.88±1.63 vs. 1.89±1.22,t = 2.909,P= 0.005), but Ang-1 only showed a slight increase (μg/L: 28.70±14.26 vs. 25.62±11.95,t = 1.005,P = 0.318). The results of Pearson correlation analysis showed that Ang-2 level was negatively correlated with forced expiratory volume in 1 second (FVC1) and the percentage of carbon monoxide diffusing capacity accounting for the expected value (DLCO%;r value was -0.227 and -0.206, andP value was 0.147 and 0.253, respectively), but no significant correlation between the level of Ang-1 and FVC1 as well as DLCO% was found (r value was -0.153 and -0.121, andP value was 0.147 and 0.253, respectively). Cox proportional hazard regression model analysis showed that the prognosis of patients with IPF was significantly affected by smoking time and Ang-2 (bothP< 0.05), and the influence of Ang-2 was greater [relative risk (RR): 1.236 vs. 1.006, P= 0.037]. Age, gender, smoking and the levels of FVC1, DLCO% and Ang-1 had no significant effect on the prognosis of IPF patients (allP> 0.05). Prognostic analysis showed that the area under ROC curve (AUC) of Ang-2 for predicting prognosis of patients with IPF was 0.692, and the best diagnostic point was 0.35μg/L, the sensitivity was 61.8%, the specificity was 73.3%, the positive predictive value was 69.8%, and the negative predictive value was 65.7% which indicated that Ang-2 could predict the prognosis of patients with IPF.Conclusion Ang-2 could assess the prognosis of patients with IPF, which is expected to be used as an indicator of predicting the prognosis of patients with IPF.

Objective To investigate the genotypes of host killing genes and their single nucleotide polymorphisms (SNPs). Methods Three hundred and twenty strains of Escherichia coli that collected from the First Affiliated Hospital of Wenzhou Medical College were analyzed. The first sample ( E1 ) contains 160 strains isolated during the years from 2002 to 2003. The second sample (E2) contains 160 strains covering the years from 2008 to 2009. The plasmids of Escherichia coli were extracted by alkaline lysis method. Solexa/Illumina sequencing technology was used to sequence plasmids metagenome. Solexa Genome Analysis System and Soap programs were used to analyze gene distribution, SNPs and lineage-specific mutations. Results 11 077 768 reads were generated and 0. 045% of them can map to the reference sequences from El sample. Whereas 9 377 792 reads were generated and 0. 053% of which mapped to the reference from E2 sample. There are nine host killing genes identified in the two samples, of which hok gene is the most prevalent. A total of 29 SNP sites dispersed in five genes of the two samples. Approximately 33% of them were non-synonymous mutations. One position of A and G is the most prevalent polymorphism. Conclusion The known nine genotypes of host killing genes were all identified in plasmids of Escherichia coli in Wenzhou. hok gene showed the highest frequency. There were SNPs in five genotypes.

Objective To study the effects of decoy strategy targeted to NF-KB on the development of trauma-associated liver inflammation in rats. Methods In this study, 108 Wistar rats were randomized into 3 groups: control group, traumatic inflammation group and traumatic inflammation plus decoy ODN group. Rats were sacrificed on 3,6,12,24,48 and 72hrs in each group respectively. Liver functions and structural changes were examined and compared between the groups. DNA binding activity of NF-KB in liver tissue was measured by EMSA. TNF-α and IL-6 gene expressin in liver tissue was assessed by RT-PCR and TNF-α and IL-6 protein level was determined by ELISA. Results The DNA binding activity of NF-kB in liver rose at 3 hours after induction of liver inflammation following trauma and peaked at 12 hours. Correspondingly, both the mRNA and protein levels of TNF-α and IL-6 elevated significantly, as well as the serum alanine aminotransferase level culminating at 24 hours after surgery. Hepatocytes was edematous, degeneration and necrosis, with dramatic destruction of lobular structures. All these changes were significantly inhibited with NF-KB decoy oligodeoxynucleotides. Conclusions Decoy oligodeoxynucleotides specifically inhibit the activity of NF-kB, and the release of pro-inflammatory cytokines, TNF-α and IL-6 release from the liver in response to traumatic inflammation decrease, hence the injury on liver structures and functions were alleviated.

Although the artificial intelligence diagnosis technology of traditional Chinese medicine(TCM)is constantly developing,the existing intelligent syndrome differentiation models can only output the diagnosis results and cannot display the reasoning process of TCM diagnosis.They do not have the interpretability of TCM diagnostic knowledge and are difficult to truly reflect the process of TCM's syndrome differentiation.This can be a long-standing problem that has puzzled the academic community.This article elaborates on the development process and technical characteristics of intelligent diagnosis in traditional Chinese medicine,explores multiple key technologies and bottlenecks in current research in reasoning of TCM's Syndrome Differentiation,and factors affecting diagnosis.A novel intelligent diagnosis model architecture that can infer and explain the process of TCM'ssyndrome differentiation was proposed,and the implementation process of the model and algorithm was explained,and detailed data instance analysis was conducted.This article will provide new methods for the development of intelligent syndrome differentiation of TCM and open up new research ideas for clinical decision-making assistance of TCM.

Objective To explore the effects of psychological intervention on the anxiety, compliance, discomfort and postoperative complication in patients receiving bronchoscopy.Methods Totally 200 patients who received bronchoscopy (fiberscope or electron scope) in Daqing Oilfield General Hospital between January and December 2014 were selected by convenience sampling and equally divided into an intervention group and a control group based on their patient ID number. Patients in the control group received routine nursing, while patients in the intervention group received psychological intervention on the basis of control group. Then the anxiety, compliance, discomfort and complication were compared between the two groups.Results There was statistically significant difference in the level of anxiety between the two groups (P<0.05). The intervention group had more patients who suffered mild or moderate anxiety but less who suffered severe anxiety than the control group. The compliance of the patients in the intervention group was higher than that of the patients in the control group (P<0.05). The incidence of intraoperative throat discomforts, suffocation and postoperative complications in patients in the intervention group was lower than that of patients in the control group (P<0.05). Conclusions Psychological intervention shows good effects on bronchoscopy, which can reduce patients' anxiety, enhance their compliance, minimize their discomfort, and curtail the incidence of complications, so that patients can receive bronchoscopy and recover successfully.

OBJECTIVETo investigate the drug resistance of enteric bacilli and its relation to the drug resistance gene cassette in the variable region and molecular evolution of class-I integron.

METHODSK-B assay was applied to measure the drug resistance of E.coli, E.cloacae and A.baumannii isolated against twelve antibiotics. The class-I integron and drug resistance gene cassettes in the variable region of the integron were detected by PCR and sequencing of amplification products. The molecular evolution of drug resistance genes in the class-I integrons was analyzed using Clustal X and MEGA software.

RESULTS54.2%-100% of A.baumannii isolates were resistant to the penicillin and cephem antibiotics, while E.coli and E.cloacae isolates had resistance rates of 41.6%-62.5% to cephem antibiotics. 62.5%(15/24) of E.coli, 67.9%(19/28) of E.cloacae and 83.3%(20/24) of A.baumannii isolates were positive for class-I integrons. 81.5% (44/54) of class-I integrons showed 4 different single band spectrums and the other class-I integrons displayed 3 different double band spectrums. In the drug resistance gene cassettes in variable regions of class-I integrons there were 7 types in 4 groups of drug resistance genes, including aac(6'), sad(3"), aad(2"), cat(4') and dfr (types 7, A13 and 15), which induced the resistance to aminoglycosides and sulfamido antibiotics and chloromycin. The class-I integrons in the isolates might be divided into 4 molecular evolution groups according to the diversity of dihydrofolate reductase encoding gene sequences.

CONCLUSIONThe enteric bacilli have a high drug resistance and frequently carry class-I integrons with 7 drug resistance gene cassettes which present 4 different evolutionary pathways.

Anti-Bacterial Agents ; pharmacology ; Drug Resistance, Multiple, Bacterial ; genetics ; Enterobacteriaceae ; drug effects ; genetics ; Evolution, Molecular ; Integrons ; genetics

Objective To explore the influence of the health management among coronary heart disease (CHD) patients after stent implementation. Methods A total of 32 CHD patients with postoperative stent implementation were randomly divided into experimental group ( received 6 months health management) and control group on (received routine discharge education) average and the results of education in two groups were compared and analyzed. Results After 6 months, there were (125. 9 ± 14. 9) mmHg, (72. 8 ± 8. 4) mmHg, (5. 44 ± 0. 53)mmol/ L, (3. 79 ± 1. 11) mmol/ L, (25. 4 ± 12. 4) U/ L, (63. 8 ± 7. 6) scores in systolic pressure, diastolic pressure, fasting blood-glucose, blood fat, liver function and body quality assessment in the experimental group compared with (133. 8 ± 11. 6) mmHg, (79. 4 ± 8. 3) mmHg, (7. 46 ± 2. 98) mmol/ L, (5. 22 ± 1. 33) mmol/ L, (38. 7 ± 13. 7)U/ L, (65. 8 ± 13. 3) scores in the control group (t = 2. 186, 2. 223, 2. 635, 2. 361, 1. 972, 2. 336,respectively;P < 0. 05). Conclusions After CHD patients acquired stent implementation, the health management can effectively minimize or stabilize the indexes of patient′ s blood pressure, blood glucose, blood fat, liver function and body quality, decrease recurrence of CHD, and promote the quality of life among CHD patients after stent implementation.