Dengue virus(DENV)diseases are a serious threat to human life and health,and there is no effective vaccine for DENV in China.In this study,a recombinant subunit candidate vaccine for DENV-2 was prepared.Using the ED Ⅲ gene of DENV-2 as the target gene,the recombinant eu-karyotic expression plasmid was constructed and transfected into suspension cells to express the target protein.The appropriate immune dose was screened to immunize mice and the immunogenic-ity was analyzed.The recombinant eukaryotic expression vector of DENV was successfully con-structed and transfected into HEK-293F to express the target protein,which was about 15 kDa in size.The mice were immunized with different doses of recombinant protein mixed with aluminum salt as adjuvant and the immunogenicity was analyzed.On the 35th day after the first immuniza-tion,the serum specific antibody levels of mice in the three protein immunization groups were sig-nificantly higher than those in the adjuvant group and PBS group,and the specific antibody levels of 10 μg D2EDⅢ protein group and 20 μg D2EDⅢ protein group were 1.43 and 1.56 times higher than those of 5 μg D2EDⅢ protein group,respectively.The antibody typing was biased to IgG1,in-dicating that the recombinant subunit candidate vaccine of DENV-2 prepared in this study mainly improved humoral immune response,but there was no significant difference in IgG1 antibody levels among the three protein immunization groups.Considering the immune effect and vaccine cost,the optimal target protein immunization dose per mouse was determined to be 10 μg.In this study,a recombinant subunit candidate vaccine of DENV-2 was prepared,which provides a refer-ence for the prevention of dengue virus.

Objective To analyze the clinical characteristics of C-TI-RADS 3 thyroid nodules with a diameter greater than 2 cm and explore their correlation with gender,nodule ingredient,contralateral cancer presence,diffuse echo changes,TPOAB and TGAB.Methods A retrospective analysis was made on the clinical and pathological information of 94 patients with thyroid nodules who were admitted to our department from September 2022 to March 2023.All the patients underwent cytological and/or histopathological examinations.The proportions of TBS I category,benign tumors,low-risk tumors,and malignant tumors were calculated.The proportion of TBS type Ⅰ,benign tumors,low-risk tumors,and malignant tumors was quantified.Subsequently,a comparative analysis was conducted among the benign,low-risk,and malignant groups in terms of clinical characteristics including gender distribution,nodule composition,contralateral cancer occurrence,diffuse echo changes presence,as well as TPOAB and TGAB levels.Results Seven cases in TBS I category were excluded.Among the remaining 87 cases with confirmed pathology results for nodules,there were 72 benign cases(38 cytology cases and 34 histology cases),5 low-risk thyroid tumors(2 cytology cases and 3 histology cases),10 malignant cases(8 PTC cases,1 FTC case,and 1 MTC case).There was a significant difference in nodule ingredient(cystic/solid)between different pathological types(x2=10.369,P=0.006).However,no statistical significance was found in terms of gender,diffuse echo changes,contralateral cancer presence,TPOAB or TGAB(P＞0.05).Further analysis showed that the proportion of solid component was higher in low-risk tumors than in benign nodules(x2=9.571,P=0.002).No statistical significance was found between malignant nodules and low-risk nodules(x2=2.143,P=0.143),or between malignant nodules and benign nodules(x2=2.165,P=0.141).Conclusion Although TI-RADS 3 nodules are generally considered as potentially benign according to various versions of thyroid imaging reporting and data system,malignant nodules still account for a certain proportion.Attention should be paid to thyroid nodules with a typical ultrasonic signs,such as cystic nodules,thyroid follicular tumors and medullary thyroid carcinoma.Ultrasound guided fine needle aspiration cytopathology is necessary for evaluating benign and malignant nodules.It is necessary to pay attention to unsatisfactory or undiagnosable specimens to improve the accuracy of diagnosis.

Objective:To investigate the risk factors and prognostic value of the textbook outcome (TO) in patients with advanced gastric cancer (AGC) who underwent neoadjuvant chemotherapy followed by surgical resection.Methods:This is a retrospective cohort study. A total of 253 patients with AGC who underwent neoadjuvant chemotherapy combined with gastrectomy and D2 lymphadenectomy in the Department of Gastric Surgery, Fujian Medical University Union Hospital from January 2010 to December 2019 were retrospectively included. There were 195 males and 58 females, aged (60.3±10.0) years (range: 27 to 75 years). The patients were then divided into the TO group ( n=168) and the non-TO group ( n=85). Multivariate Logistic regression was used to analyze the independent predictors of TO. Univariate and multivariate Cox analysis were used to analyze independent prognosis factors for overall survival (OS) and disease-free survival (DFS). Propensity score matching was performed to balance the TO and non-TO groups, and the Kaplan-Meier method was used to calculate survival rates and draw survival curves. Results:Among the 253 patients, 168 patients (66.4%) achieved TO. The Eastern Cooperative Oncology Group score ( OR=0.488, 95% CI: 0.278 to 0.856, P=0.012) and ypN stage ( OR=0.626, 95% CI:0.488 to 0.805, P<0.01) were independently predictive of TO. Multivariate analysis revealed that TO was an independent risk factor for both OS ( HR=0.662, 95% CI: 0.457 to 0.959, P=0.029) and DFS ( HR=0.687, 95% CI: 0.483 to 0.976, P=0.036). After matching, the 5-year OS rate (42.2% vs. 27.8%) and the 5-year DFS rate (37.5% vs. 27.8%) were significantly higher in the TO group than in the non-TO group (both P<0.05). Furthermore, patients in the non-TO group benefited significantly from postoperative chemotherapy (both P<0.05), but those in the TO group did not (both P>0.05). Conclusion:TO is an independent prognosis factor in patients undergoing neoadjuvant chemotherapy and surgery for AGC and is associated with postoperative chemotherapy benefits.

Autoimmune hepatitis(AIH)is a type of chronic hepatitis caused by the attack of hepatocytes by the autoimmune system,and with the prolongation of disease course,it may gradually progress to liver cirrhosis and even hepatocellular carcinoma.Although great achievements have been made in the understanding and treatment of AIH,its etiology and pathogenesis still remain unclear.T cells play a crucial role in the development and progression of AIH,and by focusing on follicular helper T cells,this article elaborates on the research advances in follicular helper T cells in AIH,in order to provide new ideas and strategies for the clinical treatment of AIH.

Objective:To investigate the risk factors and prognostic value of the textbook outcome (TO) in patients with advanced gastric cancer (AGC) who underwent neoadjuvant chemotherapy followed by surgical resection.Methods:This is a retrospective cohort study. A total of 253 patients with AGC who underwent neoadjuvant chemotherapy combined with gastrectomy and D2 lymphadenectomy in the Department of Gastric Surgery, Fujian Medical University Union Hospital from January 2010 to December 2019 were retrospectively included. There were 195 males and 58 females, aged (60.3±10.0) years (range: 27 to 75 years). The patients were then divided into the TO group ( n=168) and the non-TO group ( n=85). Multivariate Logistic regression was used to analyze the independent predictors of TO. Univariate and multivariate Cox analysis were used to analyze independent prognosis factors for overall survival (OS) and disease-free survival (DFS). Propensity score matching was performed to balance the TO and non-TO groups, and the Kaplan-Meier method was used to calculate survival rates and draw survival curves. Results:Among the 253 patients, 168 patients (66.4%) achieved TO. The Eastern Cooperative Oncology Group score ( OR=0.488, 95% CI: 0.278 to 0.856, P=0.012) and ypN stage ( OR=0.626, 95% CI:0.488 to 0.805, P<0.01) were independently predictive of TO. Multivariate analysis revealed that TO was an independent risk factor for both OS ( HR=0.662, 95% CI: 0.457 to 0.959, P=0.029) and DFS ( HR=0.687, 95% CI: 0.483 to 0.976, P=0.036). After matching, the 5-year OS rate (42.2% vs. 27.8%) and the 5-year DFS rate (37.5% vs. 27.8%) were significantly higher in the TO group than in the non-TO group (both P<0.05). Furthermore, patients in the non-TO group benefited significantly from postoperative chemotherapy (both P<0.05), but those in the TO group did not (both P>0.05). Conclusion:TO is an independent prognosis factor in patients undergoing neoadjuvant chemotherapy and surgery for AGC and is associated with postoperative chemotherapy benefits.

Autoimmune hepatitis(AIH)is chronic hepatitis caused by the attack of live cells by the immune system,and at present,the pathogenesis of AIH remains unclear.Inflammasomes are important components of innate immunity and are involved in a variety of pathophysiological processes.Studies have shown that inflammatory response associated with nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3)plays an important role in the pathogenesis of AIH,which mainly mediates the release of proinflammatory factors and pyroptosis,thereby participating in the pathophysiological process of AIH.Therefore,the development and progression of AIH can be delayed by inhibiting the activation of NLRP3 inflammasomes,which provides new ideas for the prevention and treatment of AIH.

Background Agitated behaviour in male patients with schizophrenia can lead to serious consequences for the individual and the society.Non-invasive drug delivery may help to rapidly control agitation and improve the overall patient satisfaction,but there are limited studies on the efficacy of olanzapine oral soluble film for the treatment of agitated behaviour in patients with schizophrenia.Objective To compare the efficacy and safety profile of olanzapine oral soluble film versus intramuscular haloperidol against agitated behaviour in male patients with schizophrenia,so as to provide a reference for the treatment of agitated behaviour.Methods From May 2022 to July 2023,a consecutive sample of schizophrenic patients(n=44)with agitated behaviour who met the Diagnostic and Statistical Manual of Mental Disorders,fifth edition(DSM-5)were recruited in the male psychiatric closed ward of the Third People's Hospital of Fuyang.Patients were assigned into two groups using the random number table method,each with 22 cases.Study group was treated with olanzapine oral soluble film 10 mg/d,while control group was given intramuscular haloperidol 8 mg/d.The severity of agitated behaviour was determined using Positive and Negative Syndrome Scale-Excited Component(PANSS-EC)and Agitation Calmness Evaluation Scale(ACES)at the baseline and after 6 h of drug administration.The treatment response rate was calculated based on the reduction of PANSS-EC score,and the drug side effects were assessed using Rating Scale for Extrapyramidal Side Effects(RSESE)and Barnes Akathisia Rating Scale(BARS)after 6 h of drug administration.Results After 6 h of drug administration,no statistical difference was reported in PANSS-EC score and ACES score between two groups(F=0.039,0.082,P>0.05),and the treatment response rate yielded no statistical difference between two groups(χ2=0.419,P=0.517),while study group reported a lower incidence rate of adverse reactions compared with control group,with statistically significant difference(P=0.031).BARS scores noted no statistical significant difference between two groups(t=0.587,P=0.561),whereas study group scored lower on RSES compared to control group,with a statistical difference(t=-7.367,P<0.01).Conclusion In the treatment of agitation in male patients with schizophrenia,the efficacy of olanzapine oral soluble film is comparable to that of intramuscular haloperidol,while the safety profile of the former is superior to that of the latter.

Objective To study the possibility of salivary microbiota model to diagnose laryngopharyngeal re-flux(LPR).Methods A case-control study was applied to enroll 34 patients as case group who showed significant efficacy after 8 weeks of proton pump inhibitor treatment from February 2022 to November 2022.And 47 healthy volunteers matched by age,gender and body mass index with the case group were enrolled as the control group.Their salivary samples were collected before medication,and the salivary microbiota was detected by 16S rDNA se-quencing.Bioinformatics analysis was conducted on the sequencing results to compare species differences at the ge-nus level.A total of 24 patients and 33 cases in the control group were selected as train set and the rest as test set.Random forest method was used to classify data and ten fold cross validation was applied to select the optimal bacte-rial genus combination to construct a diagnostic model.The probability of disease(POD)index was calculated and receiver operating characteristic curve(ROC)was used to evaluate the diagnostic model in diagnosis of LPR.SPSS 18.0 software was utilized for statistical analysis.Results Compared with the control group,there was a statistical difference in the relative abundance of 22 genera in saliva between the case group and the control group(P＜0.05).A diagnostic model consisting of 6 genera was constructed,namely Lactobacillus,Novosphingobium,Bacillus,Pseudoalteromonas,Ralstonia and Phocaeicola.The area under the ROC curve of the test set was 0.843,the sensi-tivity of the diagnostic model was 60.0％,the specificity was 87.71％,and the Kappa value was 0.470.Conclusion The bacterial combination diagnostic model constructed from saliva microbiota based on microbiome and machine learning can effectively distinguish LPR patients from healthy individuals,which has potential clinical application value.

In order to prepare monoclonal antibodies with blocking activity against feline TNF-α,this study successfully constructed,expressed and purified the recombinant plasmid pET-28a-sTNFα based on the soluble feline TNF-α(sTNFα)gene,and further investigated the induced ex-pression.The conditions were explored and optimized to identify its biological activity;secondly,the feline TNF-α recombinant protein was used as an immunogen for mouse immunization,after cell fusion,screening of blocking active hybridoma cells and ascites preparation,the obtained mon-oclonal antibodies were tested.The results showed that the pET-28a-sTNFα plasmid was success-fully constructed and the bioactive feline TNF-α recombinant protein was expressed in E.coli sys-tem.The molecular weight was 34 kDa and the 50％inhibitory concentration was 1.22 pg/L.Three monoclonal antibodies(A6-B7-9,H5-E2-94 and C8-A10-100)with blocking activity were success-fully screened out.The results of Western blot showed that all the three mAbs could specifically bind to TNF-α with a titer of 1:512 000.When the concentration of the three mAbs was 100 mg/L,the inhibitory effect on TNF-α was the strongest.In this study,we screened antibodies that can block the activity of cat TNF-α,in order to provide novel,safe and effective candidate drugs for the treatment of TNF-α mediated diseases in cats.

Objective To analyze the correlation between anxiety symptoms and abnormal brain function in patients with chronic obstructive pulmonary disease (COPD) under long-term chronic hypoxia. Methods Twenty-one patients with COPD complicated with anxiety were prospectively selected as COPD group, and 26 healthy individuals matched for gender and age were selected as control group. Both groups underwent high-resolution 3D-T1-weighted imaging (3D-T1WI), T2-fluid-attenuated inversion recovery (T2-FLAIR), and blood oxygen level dependent (BOLD) sequence examination. DPARSF and SPM8 software were used to analyze the amplitude of low-frequency fluctuations (ALFF) in the brain of the two groups. Results In the COPD group, the ALFF value in the left parahippocampal gyrus-cingulate gyrus increased, and the ALFF value in the right superior frontal gyrus decreased (P < 0.05). Pearson correlation analysis found that there was a negative correlation between the ALFF value in the right superior frontal gyrus and the anxiety score (r=-0.485, P=0.03). Conclusion Chronic hypoxic patients with COPD have brain functional impairment in the right superior frontal gyrus, and the degree of impairment is positively correlated with anxiety symptoms. There may also be compensatory enhancement of brain function activity in the parahippocampal gyrus-cingulate gyrus.