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 .

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 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.

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 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.

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.

Objective: To explore the differences in the gut microbiota of primary school students with different levels of sugar sweetened beverage intake, so as to provide scientific evidence for better identification of health risks in children and the development of targeted health policies. Methods: In June 2022, a total of 192 healthy primary school students from Chengdu were selected using a stratified cluster random sampling method. The sugar sweetened beverage intake was assessed through a dietary frequency questionnaire. Based on the median daily sugar sweetened beverage intake, primary school students were categorized into a low intake group ( n =96) and a high intake group ( n =96). The gut microbiota in fresh fecal samples from the two groups of primary school students was analyzed using 16S rRNA high throughput sequencing, and the diversity and community structure differences in the gut microbiota were compared. Results: Children in the low intake group had a sugar sweetened beverage intake of (21.3±1.6) mL/d, while the high intake group had an intake of (269.6±37.3) mL/d. Diversity analysis results showed that there were no statistically significant differences between the low intake and the high intake group in terms of α diversity metrics: Observed_otus index [298.50 (259.75, 342.25), 305.50 (244.25, 367.75)], Goods_coverage index [1.00 (1.00, 1.00), 1.00 (1.00, 1.00)], Chao index [304.18 (260.75, 348.78), 305.88 (245.68, 370.88)], Shannon index [5.88 (5.29, 6.45), 5.71 (4.89, 6.28)] and Simpson index [0.95 (0.91, 0.97), 0.94 (0.88, 0.97)] ( Z =-0.64, -0.76, -0.54, -1.76, -1.67, P >0.05). Furthermore, no statistically significant difference was observed in β diversity between the two groups ( R 2=0.006, P >0.05). At the genus level, the abundance of Blautia [0.033 (0.018, 0.055)] and Fusicatenibacter [0.009 (0.005, 0.015)] were higher in the low intake group compared to the high intake group [0.024 (0.013, 0.041),0.006 (0.003, 0.011)]and differences were statistically significant ( Z =-2.52, -2.81, P <0.05). LEfSe analysis highlighted intergroup differences primarily in Blautia, Fusicatenibacter and Sarcina( LDA= 3.56,3.12,3.53, P <0.05). Conclusions There is no significant difference in the diversity and overall structure of the gut microbiota in primary school students with different levels of sugar sweetened beverage intake. However, there are species variations at the genus level. The information can serve as a scientific basis for identifying health risks in primary school students and formulating targeted health strategies.

Objective: To investigate the prospective effects of intake of each food group on the development of lung function of pupils,so as to provide theoretical basis for promoting the healthy development of lung function and preventing chronic respiratory diseases in Chinese children. Methods: A cluster stratified sampling method was used to select a total of 893 pupils in grades 2-5 from Chengdu in November 2021. Dietary data of respondents were collected using a food frequency questionnaire within the past year,then the food group intake was categorized into T1, T2 and T3 from low to high by the trichotomous method, and anthropometric measurements including lung capacity were obtained in 2022. Logistic regression models and test for trend were used to analyze the prospective effects of intake of each food group on lung function development of pupils. Results: Among male students, consumption of vegetables [118.6(50.5, 188.2)g/d] and milk and dairy products [200.0(73.3, 250.0)g/d] were higher in the excellent lung capacity group than in the non excellent lung capacity group [90.0(37.1, 192.9), and 178.6(35.7, 250.0)g/d],with statistically significant differences ( Z =-1.98, -2.24); among girls, the group with excellent lung capacity consumed less staple food [391.1(273.6, 511.4)g/d] than the group with non excellent lung capacity [407.4(309.5, 594.3)g/d], and the group with excellent lung capacity consumed more aquatic products [31.2(14.6, 69.8)g/d] and milk and dairy products [215.0(107.1, 250.1) g/d ] than that of the non excellent lung capacity [19.4(10.7, 58.3), 114.3(35.7, 250.0)g/d] ( Z =-2.01, -3.33, -5.10)( P < 0.05 ). After adjusting for energy, body mass index Z score(BMI Z ), mother s education level, averge family income monthly, whether presence of smokers in the living environment, and whether participation in physical activities during the past week, among male students, T3 group of vegetable intake ( OR =0.48, 95% CI = 0.27-0.86), T2 group of bean and soy product intake ( OR = 0.52 , 95% CI =0.27-0.96)，T2 and T3 groups of milk and dairy products intake (T2： OR =0.54, 95% CI =0.31-0.93; T3： OR = 0.52 , 95% CI =0.30-0.90) were negatively associated with non excellent lung capacity ( P <0.05). Among girls, T3 group of aquatic product intake( OR =0.52, 95% CI =0.28-0.97), T2 and T3 groups of milk and dairy product (T2: OR =0.44, 95% CI =0.25- 0.76 ;T3: OR =0.33, 95% CI =0.19-0.59) were negatively associated with nonexcellent lung capacity, whereas the T2 group of red meat intake ( OR =2.51, 95% CI =1.37-4.67) was positively associated with non excellent lung capacity. Non excellent lung capacity was found to be negatively associated with vegetable and milk and dairy product intake in boys by test for trend; in girls, milk and dairy products intake was negatively associated with non excellent lung capacity, whereas red meat intake was positively associated with non excellent lung capacity ( t =-1.13，-0.44；-3.03，1.95, P trend <0.05). Conclusions Milk and dariy products intakes reduce the risk of non excellent lung capacity in pupils, vegetables intakes reduce the risk of non excellent lung capacity in boys, and the intake of red meat increases the risk of non excellent lung capacity in girls. Promoting rational food choices is necessary for children to improve healthy lung development.