Influenza A virus is an enveloped virus that belongs to the Orthomyxoviridae family. It has 8 negative RNA segments that encode 16 viral proteins. The viral polymerase consists of 3 proteins (PB 1, PB2 and PA) which plays an important role in the transcription and replication of the influenza A virus. Polymerase basic protein 1 (PB 1) is a critical member of viral polymerase complex. In order to further study the function of PB1, we need to prepare the PB1 antibody with good quality. Therefore, we amplified PB1 conserved region (nt1648-2265) by PCR and cloned it into pET-30a vector, and transformed into Escherichia coli BL2 1. The expression of His tagged PB 1 protein was induced by IPTG, and His-PB 1 proteins were purified by Ni-NTA resin. For preparation of PB 1 protein antiserum, rabbits were immunized with His-PB 1 fusion protein 3 times. Then the titer of PB 1 polyclonal antibody was measured by indirect ELISA. The antibody was purified by membrane affinity purification and subjected to immunoblotting analysis. Data showed that PB1 antibody can recognize PB 1 protein from WSN virus infected or pCMV FLAG-PB 1 transfected cells. Meanwhile, PB 1 antibody can also recognize specifically other subtype strains of influenza A virus such as H9N2 and H3N2. PB 1 polyclonal antibody we generated will be a useful tool to study the biological function of PB1.
Animals ; Antibodies, Viral ; biosynthesis ; Cloning, Molecular ; Enzyme-Linked Immunosorbent Assay ; Escherichia coli ; metabolism ; Genetic Vectors ; Influenza A Virus, H3N2 Subtype ; Influenza A Virus, H9N2 Subtype ; Plasmids ; Rabbits ; Viral Proteins ; immunology

Objective To investigate the expression and clinical significance of RegⅣand EGFR,PI3K proteins in gastric adenocarcinoma .Methods S-P immunohistochemistry was used to detect the expression of RegⅣand EGFR,PI3K proteins in pathological tissues of 73 cases with gastric adenocarcinoma and tumor -adja-cent normal gastric tissues .Results The positive expression rates of RegⅣand EGFR,PI3K proteins in 73 cases with gastric adenocarcinoma tissue were 50.7%(37/73),56.2%(41/73) and 69.9%(51/73) respectively, which were significantly different from the positive expression rates of tumor -adjacent normal gastric tissues ,be-ing 20.5%(15/73),19.2%(14/73),and 21.9%(16/73),respectively(P<0.05).The expression of RegⅣprotein was significantly correlated to differentiation degree ( P<0.05 ) and the expression of EGFR protein was significantly correlated to infiltrative depth,TNM stage,and lymph node metastasis(P<0.05).The expression of PI3K protein was significantly correlated to differentiation degree ,infiltrative depth,TNM stage and lymph node metastasis(P<0.05).There was positive correlation between the expressions of RegⅣ/EGFR,RegⅣ/PI3K and EGFR/PI3K proteins in gastric adenocarcinoma and the values of Spearman coefficient correlation were 0.325, 0.403 and 0.384,respectively(P<0.05).Conclusion RegⅣmay play an important role in gastric adenocarci-noma genesis and progression by activating EGFR /PI3K/Akt signaling pathway .

Objective To assess the clinical value of serum uric acid levels in patients with acute ST-elevation myocardial infarction(STEMI).Methods A total of 502 consecutive patients with STEMI were enrolled from January 2005 to December 2010 for retrospective study.The level of serum lipid,echocardiographic data and in-hospital major adverse cardiovascular events(MACE)in patients with hyperuricemia(n =119)were compared with those in patients without hyperuricemia(n =383).The relationship between the level of serum uric acid and the degree of diseased coronary artery was analyzed.All data were analyzed with SPSS version 17.0 software to make t test and x2 test and Pearson correlation analysis.Results Serum uric acid level was positively correlated with serum triglyceride level.Compared with non-hyperuricemia patients,hyperlipidemia was more commonly found among hyperuricemia patients (43.7％ vs.33.7％,P =0.047),and serum triglyceride level was significantly higher in hyperuricemia patients[(2.11 ± 1.24)mmol/L vs.(1.78 ± 1.38)mmol/L,P =0.014].But a significant association between serum uric acid level and one or more diseased vessels was not observed(P ≥ 0.05).Leftventricular end-diastolic diameter(LVEDd)was found to be larger in hyperuricemia patients than non-hyperuricemia patients[(53.52 ±6.19)mm vs.(52.18 ±4.89)mm,P =0.041].Higher incidence in left systolic dysfunction and diastolic dysfunction was discovered in hyperuricemia patients(36.4％ vs.15.1％,P ＜0.01; 68.2％ vs.55.8％,P =0.023).Also,hyp-eruricemia patients had more in-hospital MACE(P ＜ 0.05).Conclusions Serum uric acid level is positively correlated with serum triglyceride level,but not with the severity of coronary artery disease.Hyperuricemia patients after STEMI tended to have higher incidence in left systolic dysfunction and diastolic dysfunction,and have more in-hospital MACE.

ObjectiveTo study the effect of group interpersonal psychotherapy (IPT-G) on depression disorder of college students abused in childhood.Methods 28 college students meeting criteria of DSM-Ⅳ for depression disorder were divided into two groups:the treatment group( 14 patients)and the control group (14 patients).The treatment group was treated with IPT-G.Depression Symptom Checklist and Global Assessment Function(GAF) were measured in two groups before and after treatment.Results ①Patients in treatment group had better efficacy than those in control group( 100％ vs 57.1％,x2 =7.636,P ＜ 0.01 ).②The treatment factor and time factor had main effect to depression symptom and GAF(F =4.23 ～ 184.93,P ＜ 0.05 ),also had interaction effect(F=15.87 ～82.36,P ＜ 0.01 ).Time factor and abuse factor had main effect to depression symptom and GAF(F =7.44 ～ 183.9,P ＜ 0.05 ),but no interaction effect (F =0.01 ～ 4.72,P ＞ 0.05 ).③IPT-G,depression symptom and quality of life before treatment,age and only children had remarkable predictable functions on depression symptom(P ＜ 0.05).IPT-G had striking predictable functions on GAF (P ＜ 0.01 ).ConclusionIPT-G can improve depression symptom,mental and Social Function,while childhood abuse has no effect.

Objective To compare the disease activity score (DSA) 28-CRP and DAS28-ESR in patients with rheumatoid arthritis.Methods Two hundred and twenty-two patients were enrolled,and their sex,age,disease duration,swollen joint count,tender joint count,CRP,ESR,visual analogue scale were recorded.DAS28-ESR and DAS28-CRP were calculated and then analyzed by t test and Pearson's correlation test.Results There was a significant linear correlation between DAS28-ESR and DAS28-CRP (P＜0.05),with correlation coefficient of 0.968.Both DAS28-CRP (3.3±1.7) and DAS28-ESR (3.9±1.8) scores presented with normal distribution (P＞0.05),with the peak of the DAS28-CRP left to that of the DAS28-ESR.There was statistically significant difference between these two (P＜0.05).The difference between DAS28-CRP and DAS28-ESR was much higher in female (0.59±0.43) than in male (0.24±0.45,P＜0.05).The difference between DAS28-CRP and DAS28-ESR was not related to age and disease duration.Conclusion Attention should be paid to the assessment score when making the plan of treating to target since there is difference between DAS-28-ESR and DAS-28-CRP.

BACKGROUND:There is lack of effective diagnostic methods for early knee osteoarthritis. Proteomics refers to the large-scale experimental analysis of proteomes from the overal level of intracel ular protein compositioon, expression and modification, based on which, we can understand protein-protein interaction and relationship, thereby revealing protein functions and cel activity patterns. OBJECTIVE:To apply the isobaric tags for relative and absolute quantitation (iTRAQ) to identify proteins differential y in serums of osteoarthritis Kel gren-Lawrence classification at each stage, and to find out potential molecular markers at each stage of osteoarthritis. METHODS:Sixty patients with knee osteoarthritis were enrol ed, and according to Kel gren-Lawrence (K-L) grading, they were subdivided into K-L 0, II and IV groups. There were 10 males and 10 females randomly selected in each group. Proteins differential y regulated in serums were identified by the stable isotopes 116, 117, 118 of the iTRAQ labeled, reversed-phase column separation, mass spectrometry and Swissport database search, after the serums were subjected to high abundance proteins depletion. Final y, we analyzed the proteins identified by using bioinformatics software. RESULTS AND CONCLUSION:Total y 169 proteins were identified through iTRAQ peptides experiments of different sample tags, Q-star mass spectrum identification and MASCOT library search. 153 proteins were identified as K-L 0 to K-L IV, 153 proteins were identified as K-L 0 to K-L II, and 145 proteins were identified as K-L II to K-L IV. iTRAQ technology may help to identify novel serum markers for early diagnosis of knee osteoarthritis, indicating iTRAQ technology for proteomics serum biomarkers of osteoarthritis study has good application prospect.

Objective:To investigate the risk factors for overall survival in operable hepatocellular carcinoma with portal vein tumor thrombus (PVTT-HCC) patients and establish a scoring system.Methods:Survival data in 253 PVTT-HCC patients were retrospectively analyzed in Guangxi Medical University Affiliated Tumor Hospital. Survival curves were analyzed using the Kaplan-Meier method and log-rank test. Cox stepwise regression analysis was used to identify independent preoperative risk factors affecting overall survival. A prognostic scoring system based on independent risk factors and their relative coefficients was established to screen patients with greater hepatic resection benefits, and the identification ability of the model was based on ROC.Results:A total of 253 patients with PVTT-HCC were enrolled in this study, there were 222 males and 31 females, with a median age 44 years. The median survival time in all patients was (13.00±2.15) months. Rate of overall survival was 51.8% at 1 year, 25.0% at 3 years and 17.7% at 5 years. Multivariable Cox regression analyses showed four risk factors including: AST≥40 U/L, ALP (≥80 U/L), tumor number (>1), and incomplete tumor capsule. A prognostic scoring system was established based on these variables. The area under curve of the scoring system was 0.780 (95% CI: 0.715-0.845). Patients were classified as low- or high-risk group for hepatic resection depending on whether their score was <3 ( n=77) or ≥3 ( n=176), respectively. High-risk patients had a median survival of 10 months, compared to 29 months in low-risk patients. Low-risk patients also had better survival rates at 1 year (75.3% vs 41.5%), 3 years (47.6% vs 15.2%), and 5 years (34.7% vs 10.5%), P<0.05. Conclusion:A prognostic scoring system for hepatic resection in PVTT-HCC patients has been developed based entirely on preoperative variables. Using this system, patients belong to the low risk group have better prognosis after surgery, which can provide a basis for surgical treatment of PVTT-HCC patients.

Objective:To investigate the application effect of workshop in pediatric intravenous indwelling needle training for nursing undergraduates.Methods:A total of 214 undergraduate nursing interns were randomly divided into experimental group ( n=106) and control group ( n=108). Workshop teaching model was used in the experimental group, while the traditional teaching model in the control group. Clinical Decision Making in Nursing Scale (CDMNS), General Self-efficacy Scale (GSES) and theoretical assessment of the two groups were compared respectively before and after training, and operation evaluation was performed after training. Results:Scores of GSES, theory and operation evaluation in the experimental group were all higher than those in the control group with statistically significant differences ( P<0.05). The scores of clinical decision-making awareness was higher in the experimental group than in the control group, but the difference was not statistically significant ( P>0.05). Conclusion:Application of workshop teaching model in the training of pediatric patients with intravenous indwelling needles could enhance nursing students' self-efficacy, theoretical and operational achievements, thus improve the teaching quality.

Objectives: To evaluate the tumor-infiltrating PD-1 positive lymphocytes and regulatory T cells (Treg) as prognosis predic-tors of metastatic renal cell carcinoma (mRCC), and investigate the expression of PD-1 and Treg cells in mRCC and elucidate their corre-lation with clinicopathological parameters and prognosis. Methods: A total of 269 mRCC patients from June 2007 to June 2017 in Zhongshan Hospital, Fudan University, were included in the study. The expression of PD-1 and Tregs in mRCC samples were detected by immunohistochemistry. The relationship between the expression of PD-1 and Tregs was analyzed. Results: PD-1 positive expression in mRCC was 31.60% (85/269) and it was positively correlated with the tumor Fuhrman grade and negatively correlated with progno-sis. Tumor infiltration of Tregs in mRCC was 36.80% (99/269), and it was also positively correlated with the tumor Fuhrman grade and negatively correlated with prognosis. Univariate analysis showed that PD-1 positive lymphocytes and high Treg infiltration numbers were negatively correlated with overall survival (OS) and progression free survival (PFS) rates. Thus, PD-1 positive lymphocytes and high Treg infiltration numbers are independent prognostic indicators of OS and PFS and when combined, they can render a better pre-diction for prognosis. Conclusions: Intra-tumoral infiltration of PD-1 positive lymphocytes and Tregs can be used as significant prognos-tic indicators of mRCC, and the combined predictive effect is better than the individual predictive effect. Therefore, evaluating the number of PD-1 positive lymphocytes or infiltrating Tregs in mRCC is helpful in clinically estimating mRCC prognosis in patients.

Objective To evaluate the effect and mechanism of tribulus terrestris in kidney calcium oxalate stone in rats.Methods Sisty healthy male Wistar rats were divided into control group,model group,low-dose prevention group,high-dose prevention group,low-dose treatment group,high-dose treatment group by random number table method,each group of 10 rats.The control group was had free access to water,the model group was given 1％ ethylene glycol had free access to water,and the low-dose prevention group was given 1％ ethylene glycol had free access to water while was added in Tribulus terrestris extract 250 mg/(kg · d) treated by gavage,the high-dose prevention group was given 1％ ethylene glycol had free access to water while was added in Tribulus terrestris extract 500 mg/(kg · d) treated by gavage.The low-dose treatment group received 1％ ethylene glycol had free access to water while was added in tribulus terrestris extract 250 mg/(kg · d) treated by gavage on the 15th day.The high-dose treatment group received 1％ ethylene glycol had free access to water while was added in tribulus terrestris extract 500 mg/(kg · d) treated by gavage on the 15th day.At the end of the experiment on the 28th day,the body weight,blood urea nitrogen,creatinine,P,Ca2+,Mg2+ content,24 h urine volume,24 h urine Ca2+,Mg2 +,P,oxalic acid secretion,renal tissue homogenate detection of superoxide dismutase (SOD),glutathione peroxidase (GPX),malondialdehyde content of each group were detected.The crystal deposition and pathological changes of calcium oxalate were observed under the microscope.Measurement data were expressed as mean ± standard deviation (Mean ± SD),multiple comparisons between groups were performed using LSD-t test,and comparison between groups was performed by one-way anova analysis of variance.Results Compared with the control group,the rats in the model group had blood urea nitrogen (28.93 ±3.37) mmol/L,creatinine (162.31 ± 34.08) μmol/L,24 h uric acid (132.67 ± 27.96) μmol,Ca2+ (18.24 ± 1.94) μmol/24 h,the content of malondialdehyde (191.04 ± 20.56) nmol/mgprot in kidney homogenate was significantly increased,blood Ca2+ (2.01 ±0.28) mmol/L,renal GPX (479.82 ± 136.97) μmol/gprot,SOD(1.54 ±0.21) U/mgprot decreased significantly,these differences were statistically significant (P ＜ 0.05).Renal histopathology showed low-dose and high-dose prevention and treatment groups kidney calcium oxalate crystals and kidney damage were less than the model group.Conclusion The extract of Tribulus terrestris has a beneficial effect against calcium oxalate urinary stone formation induced by ethylene glycol.