Objective:To analyze the molecular evolutional characteristics of the hemagglutinin and neuraminidase genes of influenza A (H3N2) viruses isolated in Yancheng from 2022 to 2024.Methods:The throat swab specimens of influenza-like illness ( ILI) from sentinel surveillance hospital and outbreak sites were detected using the method of real time Rt-qPCR. The influenza A(H3N2) viruses were isolated using MDCK cells culture method from April 2022 to Marh 2024. The strains isolated from 2022 to 2024 were selected randomly and their sequences of the HA1 and NA genes were amplified through one step RT-PCR method and the PCR products were sequenced.The nucleotide and amino acid site variations and evolutionary characteristics of the genes were analyzed using relevant bioinformatics software. The mutations of genes and nucleic acid locus were analyzed and the evolutional trees were generated using bioinformatics software.Results:A total of 5 020 samples were collected between April 2022 and March 2024, the positive detection rate of influenza virus nucleic acid was 18.59%(933/5 020).The winter and spring influenza peaks were obvious in the two monitoring seasons from April 2022 to March 2024. Among them, the summer influenza peak was obvious in the monitoring season from April 2022 to March 2023, and the H3N2 subtype influenza virus was the dominant epidemic strain in the two monitoring seasons. Genetic evolution tree displayed: the clustering relationships of the respective branches of HA1 and NA genes of 32 strains isolated in Yancheng were basically the same.The HA1 and NA genes of 24 strains isolated from 2023-2024 in Yancheng and the 2022-2024 Northern Hemisphere vaccine strain A/Darwin/9/2021 (H3N2) were located in the 3C.2a1b2a.2a.3a.1 evolutionary lineage, while the 8 strains isolated in the 2022 in Yancheng and the 2021-2022 Northern Hemisphere vaccine strain A/Cambodia/e0826360/2020 (H3N2) were located in the 3C.2a1b.2a.1a evolutionary lineage.The 6 strains (A/JSTH/11735/2023, A/JSTH/11788/2023, A/JSTH/11974/2023, A/JSYD/353/2023, A/JSYD/354/2023, A/JSTH/138/2023) all exhibited variations in the F79L, N122D, P239S, and K276E amino acid sites, which were present in both sporadic and outbreak strains. Because the strains of the antigen epitopes, receptor binding sites and glycosylation sites in the HA1 genes had a certain degree of variations in Yancheng in the 2022-2024 year, the immunogenicity matching between the 24 strains isolated in the 2023-2024 and the Northern Hemisphere vaccine strain A/Darwin/9/2021 was good, while the immunogenicity matching between the 8 strains isolated in the 2022 and the Northern Hemisphere vaccine strain A/Cambodia/e0826360/2022 was good; 32 strains isolated from 2022 to 2024 had no mutations in catalytic residues and drug resistant sites of NA genes.Conclusion:These result indicated that the HA1 and NA genes of influenza A/H3N2 viruses circulated in Yancheng city from 2022 to 2024 are changed gradually.The accumulation of these mutations would result in antigenic drift of influenza A(H3N2) viruses and increase the mismatching of the recommended vaccine strain.Compared with the vaccine strain A/Darwin/9/2021(H3N2), the strains isolated in the 2022 had substantially result in antigenic drift on the whole.The influenza A(H3N2) viruses surveillance should be strengthened to find the new mutant of virus in time.

BackgroundBeing complex and highly heterogeneous with regard to the etiology and clinical manifestations of depression， neuroimaging studies make a breakthrough for exploring the biological subtypes of depression， while the current data-driven approach for the identification of subtyping depression using structural magnetic resonance imaging （MRI） data is insufficient. ObjectiveTo explore the biological subtypes of depression using diffusion tensor imaging （DTI） and machine learning methods. MethodsA total of 127 patients with depression who attended Beijing Anding Hospital from September 2017 to August 2021 and met the Diagnostic and Statistical Manual of Mental Disorders， fourth edition （DSM-IV） diagnostic criteria were included， and another 80 healthy individuals matched for gender and age were recruited through advertisements in surrounding communities during the same period. DTI findings， demographic characteristics and clinical data were collected from all participants. Tract-based spatial statistics （TBSS） and the Johns Hopkins University （JHU） white matter probability maps were used to extract fractional anisotropy （FA） values of white matter tracts. A semi-supervised machine learning technique was used to identify the subtypes， and the FA values for whole brain white matter of patients and controls were compared. ResultsPatients with depression were classified into two biological subtypes. FA values in multiple tracts including corpus callosum and corona radiata of subtype I patients were smaller than those of healthy controls （P<0.01， FDR corrected）， and FA values in middle cerebellar peduncle， left superior cerebellar peduncle and left cerebral peduncle of subtype II patients were larger than those of healthy controls （P<0.01， FDR-corrected）. Baseline Hamilton Depression Scale-17 item （HAMD-17） score yielded no statistical difference between subtype I and subtype II patients （P>0.05）， while subtype I patients scored lower on HAMD-17 than subtype II patients after 12 weeks of treatment （t=2.410， P<0.05）. ConclusionDepression patients exhibit two biological subtypes with distinct patterns of white matter damage. Furthermore， the subtypes respond differently to the medication treatment. ［Funded by the National Key Research and Development Program of China （number， 2016YFC1307200）， the Scientific Research and Cultivation Program of Beijing Municipal Hospitals （number，PX2023066）， Beijing Anding Hospital， Capital Medical University （number，YJ201904， YJ201911）； www.chictr.org.cn number： ChiCTR-OOC-17012566］

Objective:To assess the psychometric properties of the Chinese version of 12-item immediate mood scale(IMS-12) in patients with depression.Methods:From January to June 2018, a total of 459 patients with depression recruited from an outpatient clinic by convenient sampling approach.All the subjects were assessed by the Chinese version of the IMS-12, and 43 of them were assessed again at the end of the first week.The 16-item quick inventory of depressive symptomatology (QIDS-SR16), and the generalized anxiety disorder scale-7(GAD-7) were used as validity indicator.The factor structure of the scale was evaluated by exploratory and confirmatory factor analyses.The internal consistency of the Chinese version of the IMS-12 scale was evaluated by Cronbach’s alpha coefficient.The intraclass correlation coefficient (ICC) was used to evaluate test-retest reliability.Pearson’s correlation coefficient was used to evaluate calibration validity.The softwares of SAS 9.4 and Mplus 8.5 were used for statistical analysis.Results:The exploratory factor analysis indicated that the fitting result of the two-factor model was good(including depression and anxiety factors). The results of the confirmatory factor analysis indicated that the factor model fit well and met the reference standard ( χ2/ df=2.82, CFI=0.936, TLI=0.920, RMSEA=0.088). The Cronbach’s alpha coefficient of the Chinese version of the IMS-12 was 0.95, and the ICC for test-retest reliability was 0.85.The correlation coefficients of the total IMS-12 score with the QIDS-SR16 score and with the GAD-7 score were 0.69 and 0.70, respectively. Conclusion:The Chinese version of the IMS-12 has good reliability and validity and is suitable for the clinical assessment of depressive and anxiety symptoms in patients with depression.

Objective:To explore the application value of metagenomics sequencing in the etiological diagnosis of bacterial meningitis(BM) in children and improve the diagnosis and treatment effect of BM in children.Methods:All BM cases were collected from Jiangxi Provincial Children′s Hospital from February 1, 2018 to January 31, 2020.Meanwhile, such biological samples as blood and cerebrospinal fluid were collected for traditional etiological testing and metagenomics sequencing.The results of traditional etiological testing were regarded as the gold standard, and the specificity and sensitivity of metagenomics sequencing in the diagnosis of BM in children were confirmed.Results:A total of 45 cases were collected in this study, including 31 males and 14 females, with the age between (74.74 ± 58.67) months.Twenty-six cases were identified by metagenomics sequencing, with the positive rate being 57.78%.Among them, there were 8 cases of Streptococcus pneumoniae, 2 cases of Escherichia coli, 2 cases of Neisseria meningitidis, 2 cases of Staphylococcus, 2 cases of Salmonella, 2 cases of Mycobacterium multiplex, 1 case of Streptococcus intermedius, 1 case of Streptococcus pyogenes, 1 case of Streptococcus paris, 1 case of Streptococcus salivarius, 1 case of Haemophilus influenzae, 1 case of Pseudomonas aeruginosa, 1 case of Acinetobacter baumannii, and 1 case of Aspergillus.The traditional etiological positive rate was 17.78%, and the metagenomic next-generation sequencing (mNGS) positive rate was 57.78% ( P=0.014, kappa=0.273). As per the comparison results, the sensitivity, specificity, positive predictive value, negative predictive value, Youden index and misdiagnosis rate were 100.00%, 51.35%, 30.76%, 100.00%, 51.36%, 48.64% and 0, respectively. Conclusions:Metagenomics examination had high sensitivity, which can improve the etiological diagnosis rate of acute BM in children, especially in case of high clinical suspicion of infection.Therefore, metagenomics examination should be selected as early as possible when the etiology cannot be determined by traditional approaches.

Objective:To explore whether working memory span training can expand working memory capacity.Methods:A randomized controlled trial design was adopted and a total of 60 healthy college students were recruited and randomly divided into training group ( n=30, receiving adaptive training of spatial breadth task) and control group ( n=30, receiving non-adaptive training of low difficulty spatial breadth task). The cognitive behavior and event-related potential (ERP) data of all subjects when completing the change awareness task were collected before and after training.The SPSS 22.0 statistical software was used for data analysis. The differences between the training group and the control group before and after training were compared by repeated measurement analysis of variance. Results:Repeated measurement ANOVA showed that there were significant time and group interactions at the levels of cognitive behavior(K score, F=5.352, P=0.025) and ERP (CDA, F=4.644, P=0.037) levels. Further post test found that compared with pre-training (pre-test), the K-score ((0.51±0.93), (1.61±1.07), F=26.81, P<0.001) and CDA ((-1.49±1.07)μV, (-2.03±0.94)μV, F=4.731, P=0.041) of the training group increased significantly after training (post-test), and there was no significant difference in K-score and CDA of the control group before and after training (boh P>0.05). Conclusion:Working memory span task can be used as an effective training paradigm to improve working memory capacity.

Objective To set up a living mice colonoscopy platform to establish an orthotopic model of colorectal cancer in mice under direct vision,and to observe its biological behavior such as metastasis.Methods Eighteen-week-old male C57/BL mice were anesthetized,and the intestinal lumen of the mice was examined by a self-developed living mice colonoscopy and Olympus URF-P5 ureteroscopy,respectively.The imaging effects of the two methods were compared.Human colon cancer HT-29 cells were injected into the colonic mucosa of BALB/c-nu mice under direct vision.The colonoscopy was performed on the 3rd,7th and 15th day after the injection to observe the tumor formation in the intestinal lumen.The mice were sacrificed when the body weight decreased significantly or cachexia appeared,and then the abdominal cavity was examined including the tumor formation and metastasis.Results The self-developed living mice colonoscopy platform can provide clear vision of enteric cavity,and no mice died in the colonoscopy examination.In vivo subcutaneous injection of HT-29 cells in mice was performed with a perforation rate of 15％,a mortality rate of 33.3％,a tumor formation rate of 62.5％,an abdominal metastasis rate of 60％,a liver metastasis rate of 25％,and an abdominal wall transfer rate of 25％.Conclusion The self-developed mice colonoscopy platform can be used for the study of colorectum in living mice.The imaging effect is no less than that of Olympus URF-P5 ureteroscopy.In addition,an orthotopic colorectal cancer model can be established by this platform combing with submucosal injection technology.

Objective:To explore the effects of protoporphyrin Ⅸ (PpⅨ)-mediated photodynamic therapy (PDT) on induction of apoptosis and death in colon cancer cell and the underlying mechanisms.Methods:The cell killing effect of PDT on HCT116 cell was determined by cell counting kit (CCK).The cells were divided into a control group,a single light group,a single PpⅨ group,and a PDT group.Hoechst 33342 and flow cytometry was used to assess the cell apoptosis.Western blot was employed to analyze the expressions ofbd-2,bax,and caspase-3.Reactive oxygen species (ROS) was detected by flow cytometry.Results:The viability of HCT116 cell was decreased gradually with the increase of irradiation dose (P<0.05).Compared to the other 3 groups,ROS production,the number of apoptotic cells and the protein expressions ofbax and caspase-3 in PDT group increased,while bcl-2 expression was decreased (P<0.05).Conclusion:PpⅨ-mediated PDT can enhance the apoptosis in HCT116 cell,which may be related to mitochondrial apoptosis pathway.

Objective To explore the effect of virtual laboratory (VL) + flipped classroom in teaching of virus infection diagnosis.Methods 40 students of Class One from clinical medical undergraduates of Grade 2014 were randomly taken as the experimental group,with 40 students of Class Two as the control group.The experiment group adopted flipped classroom teaching by virtual lab platform and classroom activities,while the control group adopted traditional classroom teaching such as watching video and lecturing.Finally post-test scores were compared by the independent samples t-test of SPSS 18.0 statistical software between the two groups.The teaching effects were evaluated through questionnaires survey in experimental group.Results The scores (82.73 ± 2.62) of comprehensive assessments were superior to the control scores (57.94 ± 4.65).Difference between the two groups was statistically significant (t=29.380,P=0.000).Students' satisfaction concerning the teaching methods and effects of the flipped classroom in experimental group was up to 85％.Conclusion Flipped classroom based on internet virtual lab platform in teaching of virus infection diagnosis can enhance the teaching quality and improve students' learning enthusiasm and thinking ability.

Objective To assess the association between rs7597593 polymorphism of ZNF804A gene and schizophrenia,and to assess the relationship between rs7597593 polymorphism and working memory.Methods Schizophrenia patients and healthy controls were diagnosed in accordance with Diagnostic and Statistical Manual of Mental Disorders-Fourth Edition (DSM-Ⅳ) ; 767 schizophrenia patients and 690 healthy controls were involved.Restriction fragment length polymorphism (RFLP) was carried out to genotype rs7597593 polymorphism.The cognitive function of working memory was assessed by the N-back task.Statistical analyses were carried out with SPSS19.0 software.Results The study found no significantly different genotype frequencies (x2=1.519,P=0.468) and allele frequencies(x2=1.263,P=0.261) of rs7597593 polymorphism between schizophrenia patients and healthy controls,however in the subgroup of higher IQ (IQ ≥ 110),there were significant different distributions of both genotype and allele (x2 =9.411 and 6.529; P=0.009 and 0.011 respectively).It was also found in this subgroup that risk T allele was associated with more error at 1-back task (F=6.854,P=0.009).Conclusion These results indicated that rs7597593 polymorphism was associated with individuals having spared cognitive function; carriers of T allele had worse cognitive function,which maybe a pathway that it contributes to schizophrenia.