Objective·To investigate the evolution of sarcoma antigen 1(SAGE1),a member of the cancer-testis antigen(CTA)family,in three representative primate species—Macaca mulatta(Rhesus),Callithrix jacchus(Marmoset),and Microcebus murinus(Mouse Lemur),as well as the conservation of its interaction with INTS3,a subunit of the integrator complex.Methods·The interaction between INTS3 and SAGE1 in these primates and the interaction between INTS3 and human SAGE1 mutants were first validated in HEK293T cells by using co-immunoprecipitation(Co-IP).Truncated proteins were then tagged with His-SUMO and GST,respectively,and expressed in Escherichia coli,followed by a series of purification steps to obtain the target proteins.Surface plasmon resonance(SPR)was used to measure the binding affinity of the target proteins,and size exclusion chromatography with multi-angle light scattering(SEC-MALS)was used to determine the stoichiometry of the complex.Additionally,molecular docking was employed to predict the binding model of the truncated SAGE1 and INTS3.Mutations were performed on human SAGE1 key interface residues to analyze their binding to INTS3 by Co-IP.Results·The interaction between the full-length human-derived INTS3 and the full-length SAGE1 from the three primate species was confirmed by Co-IP.Truncated proteins were purified through multiple steps of tandom chromatography.The dissociation constants of the three pairs of truncated INTS3-SAGE1 were measured via SPR,and the SEC-MALS results demonstrated that the binding ratio of INTS3-SAGE1 was 2∶1.Furthermore,molecular docking predicted a structural model of the truncated INTS3-SAGE1 complex and the binding interface was extensively constituted with hydrophobic contacts assisted by some hydrophilic interactions.The interaction between the mutant SAGE1 and INTS3 full-length protein was significantly weakened.Conclusion·There is a conserved interaction between INTS3 and SAGE1 across Rhesus,Marmoset,and Mouse lemur.

Objective:To summarize the best evidence for external auditory canal irrigation in patients with cerumen embolism.Methods:The clinical decisions, guidelines, systematic reviews, expert consensus, group standards, evidence summaries, and randomized controlled trials regarding external auditory canal irrigation in patients with cerumen embolism were retrieved from databases and websites such as BMJ Best Practice, UpToDate, Guidelines International Network, National Institute for Health and Clinical Excellence, Joanna Briggs Institute Evidence-Based Health Care Center Database, PubMed, Embase, China National Knowledge Infrastructure, WanFang data, and China Biology Medicine disc. The search period was from database establishment to February 15, 2023. Six researchers screened the literature, evaluated the methodological quality, and extracted and summarized the best evidence for external auditory canal irrigation in patients with cerumen embolism.Results:A total of nine articles were included, including one clinical decision, two guidelines, two systematic reviews, one group standard, and three randomized controlled trials. Sixteen pieces of evidence were summarized from six aspects of operators: pre-operation evaluation and preparation, operation process, post-operation handling, health education, and adverse reactions during operation.Conclusions:This paper summarizes the best evidence for external auditory canal irrigation in patients with cerumen embolism. Medical and nursing staff should carefully select and apply evidence based on clinical scenarios and patient's wishes.

Objective:To investigate the epidemiological and genetic characteristics of human metapneumovirus (hMPV) in Tianjin during two influenza epidemic seasons from October 2020 to March 2021 and from October 2021 to March 2022, and enrich the whole genome database of hMPV in China.Methods:A total of 1 040 pharyngeal swab samples collected from patients with influenza-like illness (ILI) were analyzed using microfluidic chip fluorescence quantitative PCR. RT-PCR was used to amplify the whole genome in hMPV-positive samples, and the second-generation sequencing was performed for complete genome sequencing. Bioinformatics software including CLC, DNAStar, and MEGA was used for sequence assembly, nucleotide and amino acid homology analysis, and phylogenetic tree mapping.Results:Among the 1 040 samples, 25 were positive for hMPV with a positive rate of 2.40%. The highest positive rate was observed in the age group of 3 to 5 years, reaching 3.71% (16/431). During the influenza epidemic seasons, the detection rate of hMPV peaked in December, reaching 6.67% (12/180). Twelve strains were successfully sequenced, and there were seven of type B2, four of type A2b, and one of type B1. More variations were detected in the G gene, with 111nt-dup sequence repeats observed in the G gene of three A2b strains.Conclusions:The prevalence of hMPV peaks in December during the influenza epidemic seasons in Tianjin, with Type B2 being the predominant type. Except for the G gene with more mutations, other genes remain stable.

OBJECTIVE To investigate the effect and mechanism of dihydroartemisinin(DHA)on lipo-polysaccharide(LPS)-induced acute lung injury(ALI)in mice using whole-genome sequencing.METHODS An ALI mouse model was established via intraperitoneal injection of 10 mg·kg-1 lipopolysaccharide.The mice were divided into normal control group(n=10),model group(n=10)and model+DHA group(n=10).The mice in the model+DHA group were injected intraperitoneally with 20 mg·kg-1 DHA,while those in the normal control group and LPS group were injected intraperitoneally with solvent of DHA,saline containing 1%Tween 80 and 10%Macrogol 400.The mice were executed 24 h after drug administration.The wet and dry weight ratio(W/D)of lung tissue was calculated.Hematoxylin-eosin(HE)staining was used to observe histopathological damage in the lung.Classified counts of inflamma-tory cells in alveolar lavage fluid were performed.Enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of interleukin-1β(IL-1β),IL-6,and tumor necrosis factor-α(TNF-α)in alveolar lavage fluid.Real-time quantitative PCR(RT-qPCR)was used to detect mRNA levels of placenta-specific 8(Plac8),Toll-like receptor 7(TLR7),IL-1β,IL-6 and TNF-αin lung tissue.The whole gene transcriptome was sequenced by RNA transcriptome sequencing(RNA-seq)using the Illumina HiSeq high-throughput sequencing platform before the function and signal pathway of differentially expressed gene mRNA between the groups were enriched and analyzed using GO and KEGG enrichment analysis methods.RESULTS Compared with the model group,the lung W/D values of mice,the pathological damage,inflammatory cells in alveolar lavage fluid,expression levels of IL-1β,IL-6 and TNF-α in alveolar lavage fluid(P<0.01,P<0.01,P<0.01),and the mRNA expression levels of IL-1β,IL-6 and TNF-α were significantly reduced in lung tissues in the model+DHA group(P<0.01,P<0.05,P<0.05).Whole gene transcriptome sequencing revealed that immune-related Plac8 and TLR7 genes were significantly upregu-lated(P<0.01)in mouse lung tissue of the model group but significantly downregulated(P<0.05)in mouse lung tissue of the model+DHA group.The results of RT-qPCR of Plac8 and TLR7 verified the results of whole gene transcriptome sequencing.GO and KEGG analysis showed that Plac8 and TLR7 were mainly related to the regulation of cytokine production,T/B cell activation and signal transduction,chemo-kine signal transduction and NF-κB signal transduction.CONCLUSION DHA might reduce LPS-induced lung damage and ameliorate the inflammatory condition in lungs of ALI mice.The mechanism of action may be that DHA negatively regulates the signaling pathways involved in TLR7 and Plac8 by decreasing the expressions of TLR7 and Plac8 mRNA before regulating a series of immune responses such as secretion of inflammation-related cytokines and activation of immune cells,thereby reducing inflam-matory damage in lungs.

Objective : To investigate the effect of platelet particles on the extent of intestinal inflammation and in⁃ testinal mucosal permeability in mice with dextran sodium sulfate induced colitis. Methods : The experiment was divided into four groups : normal control group ( n = 10 , drinking sterile distilled water + intraperitoneal injection of 0. 9% sodium chloride solution) , PMPs group ( n = 10 , drinking sterile distilled water + intraperitoneal injection of PMPs) , DSS model group ( n = 10 , drinking DSS solution + intraperitoneal injection of 0. 9% sodium chloride solution) , and experimental group ( n = 15 , drinking DSS solution + intraperitoneal injection of PMPs) . Peripheral blood⁃derived PMPs suspension was collected from inflammatory bowel disease ( IBD) patients. A colitis model was constructed in mice by allowing them to freely drink a 5% DSS solution for 1 week , followed by continuous intraperitoneal injection of PMPs for 7 days. Disease activity index (DAI) scores was recorded daily and the severity of intestinal inflammation with histopathological scores (HI) was assessed by HE staining of colon samples at the end of the experiment. Myeloperoxidase (MPO) , neutrophil elastase (NE) , citrullinated histone H3 (citH3) , and free DNA levels were measured in colon homogenate , observe intestinal mucosal structure by transmission electron microscopy , and intestinal permeability was tested using fluorescein isothiocyanate⁃dextran (FITC⁃D) . Results: Compared with the normal control group , the colonic mucosa of mice in the PMPs group showed edema , severe destruction of epithelial structure , extensive aggregation of inflammatory cells , and increased overall HI score (P < 0. 01) ; the levels of inflammatory factors such as IL⁃1β and TNF⁃α in colonic tissue homogenates of mice in the PMPs group increased (P < 0. 05) , and the expression of NETs increased (P < 0. 05) ; the plasma FITC⁃D level of mice in the PMPs group significantly increased (P < 0. 05) , and the permeability of intestinal mucosa increased. Compared with the DSS group , the experimental group mice had higher plasma FITC⁃D levels ( P < 0. 05 ) and more electron microscopic colonic epithelial damage. Conclusion PMPs induces NETs formation in mice , promotes colonic inflammation in mice , increases intestinal mucosal permeability and aggravates intestinal inflammation in mice with DSS colitis.

Objective: To study the chemical constituents from the leaves and twigs of Callicarpa cathayana. Methods: The chemical constituents were isolated and purified by column chromatography on silica gel, MCI gel CHP 20P/P120, Sephadex LH-20, and HPLC. The structures of the compounds were determined by HR-ESI-MS, 1D and 2D NMR data. Results: A total of 24 compounds were isolated from the 85% methanol extract of leaves and twigs of C. cathayana. They were identified as cathayanalactone G (1), a new diterpene, and 23 known compounds as patagonic acid (2), (-)-16-hydroxycledroda-3,13-dien-16,15-olide-18-oic acid (3), 15-methoxypatagonic acid (4), oleanolic acid (5), ursolic acid (6), siaresinolic acid (7), pomolic acid (8), α-amyrin (9), tormentic acid (10), lupeol (11), 5,7-dihydroxy-3,4′-dimethoxyflavone (12), 5,4′-dihydroxy-3,7,3′-dimethoxyfla-vone (13), 5-hydroxy-3,6,7,4′- tetramethoxyflavone (14), salvigenin (15), kaemferol (16), astragalin (17), pinoresinol 4-O-β-D-glucopyranoside (18), paulownin (19), β-sitosterol (20), β-sitosterol β-D-glucopyranoside (21), 5-hydroxy-coumarin (22), isocopoletin (23), and 4-hydroxycinnamic acid (24). Conclusion: Compound 1 is a new labdane diterpene. Compounds 10, 13, 16 and 17 are isolated from the genus Callicarpa for the first time. Compounds 7, 8, 9, 12, 14, 23 and 24 are reported from C. cathayana for the first time.

Objective:To understand the epidemiological characteristics and pathogenic spectrum of influenza-like illnesses in Tianjin Children′s Hospital from October 2020 to March 2021, and to provide reference for the prevention, control and clinical diagnosis and treatment of influenza-like illnesses.Methods:A total of 520 throat swabs samples were collected from patients with influenza-like illnesses in sentinel hospitals. Thirty respiratory tract pathogens were detected by real-time fluorescence quantitative PCR. The results were statistically analyzed by descriptive epidemiological methods.Results:Among the 520 samples, 239 were positive for 16 respiratory pathogens with a positive rate of 45.96%. The top three pathogens were respiratory syncytial virus (9.62%, 50/520), rhinovirus (9.62%, 50/520) and cytomegalovirus (5.96%, 31/520). The positive rate of respiratory pathogens was 49.67% in males and 40.91% in females and the difference between males and females was statistically significant (χ 2=3.919, P<0.05). There were significant differences in the positive rates among three age groups (χ 2=6.182, P<0.05) with the highest positive rate in the <2 years old group (52.91%, 91/172) and the lowest rate in the >4 years old group (38.10%, 40/105). There were significant differences in the positive rates detected in different months (χ 2=15.358, P<0.05) and the highest detection rate was in December (58.00%, 58/100), followed by those in November (52.50%, 42/80) and January (47.50%, 38/80). The multiple infection rate was 21.76% (52/239) and most of the multiple infections were caused by rhinovirus and other pathogens (48.08%, 25/52). Conclusions:Respiratory syncytial virus, rhinovirus and cytomegalovirus were the predonimant pathogens responsible for influenza-like illnesses in Tianjin Children′s Hospital from October 2020 to March 2021. Multiple infections were common and children under 2 years old were more susceptible. The detection rate of respiratory pathogens varied in different months. It was necessary to strengthen the surveillance and research on those respiratory pathogens in order to provide scientific data for the prevention and control of respiratory diseases in children.

Objective:To investigate the sodium intake, potassium intake, sodium- potassium ratio and influencing factors of family cooks and members in six regions of China.Methods:Using the multistage random sampling method, a total of 1 576 family cooks and their family members were recruited from 6 regions in China. Questionnaire survey and physical examination were conducted to collect basic information. 24 h urine was collected to test 24 h urinary sodium and potassium levels.Results:After excluding unqualified urine samples, a total of 1 530 people were included in the study. For all participants, 24 h urine sodium value was (4.39±1.93) g, 24 h urine potassium value was (1.59±0.62) g and sodium-potassium ratio was 5.02±2.18. 24 h urinary sodium was negatively correlated with age, female, junior high school education or above, and annual family income ( P<0.05), and positively correlated with perceived salty taste, BMI and SBP ( P<0.05). The 24 h urine potassium value was negatively correlated with eating out almost everyday ( P<0.05), but positively correlated with BMI and unwillingness to reduct salt ( P<0.05). The sodium-potassium ratio was negatively correlated with age, female and junior high school education or above annual family income ( P<0.05), and positively correlated with perceived salty taste, SBP and eating out 3-5 d/week ( P<0.05). Conclusions:The 24 h urinary sodium value and sodium potassium ratio of family cooks and family members in six regions of China are still at high level, while the 24 h urine potassium value is low, with multiple factors. Therefore, it is necessary to carry out long-term comprehensive salt reduction intervention for family cooks and their members.

Objective:To examine the association of blood uric acid (UA) and cognitive impairment (CI) among oldest-old adults in China.Methods:Data was collected in 9 longevity areas of China from Healthy Aging and Biomarkers Cohort Study (HABCS) conducted during 2017-2018. Finally,1, 622 elderly aged 80 years and older with complete information on blood UA and cognitive function score were included in this study. Information on demographic characteristics, lifestyle, and health status were collected through questionnaire and physical examination. Venous blood samples of the participants were collected to test blood UA level. Cognitive impairment (CI) was assessed using the Chinese Mini-Mental State Examination (MMSE) according to personal educational level. Generalized Estimating Equations (GEE) model for binary data was used to analyze the association of blood UA and CI, and further compared the associations among different age and body mass index (BMI) groups.Results:Of the 1 622 oldest-old, the mean age was (92.2±8.1) years, 656 (40.4%) were male, the mean level of blood UA was (343.3±86.2) μmol/L, and 482 (29.7%) oldest-old had CI. Compared with the lowest quartile of UA, the risks of CI in the second, third and highest quartiles were gradually reduced, the corresponding ORs and 95% CI were 0.99 (0.71-1.33), 0.87 (0.68-0.94) and 0.69 (0.48-0.85), respectively; and the linear trend test was statistically significant ( P<0.001). Subgroup analysis showed that the effects of higher UA associated with lower risk of CI were stronger in younger oldest-old (aged 80-89 years) and thinner group (BMI<24) ( Pinteraction<0.05). Conclusions:Blood UA was negatively associated with the risk of having CI in the oldest-old among the nine longevity areas of China.

Objective:To investigate the relationship between the neutrophil-to-lymphocyte ratio (NLR) and the risk of depression symptoms among older adults aged 65 and above in 9 longevity areas of China.Methods:Data was collected in 9 longevity areas of China from Healthy Aging and Biomarkers Cohort Study (HABCS) conducted between 2017 and 2018. Finally,2018 elderly aged 65 years and above with complete information on neutrophil count, lymphocyte count and depressive symptoms were included in this study. Information on demographic characteristics, lifestyle, and health status was collected through questionnaire and physical examination. Complete blood counts which included lymphocytes and neutrophils were obtained by testing venous blood samples. Participants were divided into four groups by the quartile of NLR level, i.e. Q1, Q2, Q3, Q4. Logistic regression model was applied to analyze the association of NLR with depression symptoms. Results:Among 2 018 older adults, the mean(±SD) age was 82.6(±10.73), 1 032(51.14%) were male, 390(19.33%) were detected with depressive symptoms. Compared with participants of NLR in the 1 st quartile, the OR(95% CI) of risk for depressive symptoms was 1.47 (0.99, 2.19), 1.67 (1.13, 2.47) and 1.95 (1.32, 2.89), respectively. Conclusion:Increased NLR level is significantly related to depressive symptoms among elderly aged 65 years and above in 9 longevity areas in China.