Objective: To optimize the technology and process for recovering immune cells from discarded leukocyte filter through reverse flushing, and to investigate the effects of different reverse flushing solutions and physical conditions (temperature, soaking, and ultrasound) on the cell recovery rate. Additionally, to evaluate the feasibility and quality of isolating and culturing NK cells from the recovered cells. Methods: Normal saline, PBS, and PBS-EDTA were used as reverse flushing solutions to backwash the filter, and the white blood cell recovery rates were compared. The reverse flushing efficiency was optimized by adjusting the temperature of the reverse flushing solution, filter soaking treatment and ultrasonic treatment. The Ficoll separation method was employed to isolate peripheral blood mononuclear cells (PBMCs) from the elution solution and conduct in vitro expansion and culture. Lymphocyte subsets were identified and quantified using flow cytometry. Results: The physiological saline served as the backflush fluid was the most stable. The reverse temperature had a significant impact on the recovery rate of white blood cell subgroups. The recovery rates of lymphocytes and monocytes at 37℃ were significantly higher than those at 4℃ [(35.56±5.55)% vs (22.88±0.31)%, (32.42±4.82)% vs (8.00±1.6)%, t=5.581 and 5.758, respectively, both P<0.001)], and at 25℃ [(35.56±5.55)% vs (22.42±6.13)%, t=5.904, P<0.001; (32.4±4.82)% vs (21.11±1.23)%, t=2.629, P=0.030)]. The recovery rates of white blood cell subgroups after 37℃ immersion were higher than those after direct reverse flow at 37℃, but the difference was not statistically significant (P>0.05). The recovery rate of cells after ultrasonic immersion at 25℃ was further increased compared to that after immersion at 25℃, but the difference was not statistically significant (P>0.05). The purity of the NK cells obtained through sorting reached (94.2±4.5)%, and the viability was (94.8±0.9)%. After 15 days of culture, the cell count was expanded approximately 450 times, yielding more than 10 /mL highly active NK cells. Conclusion: By optimizing the backflushing process, immune cells can be efficiently recovered from discarded filters. The 37℃ backflow liquid has the best backflow effect; soaking and ultrasonic treatment can be used as auxiliary methods to improve recovery rates. The NK cells from this source have high purity, good activity, and strong expansion ability, and can be used as a reliable cell source for immunotherapy research, and in line with the circular economy concept of resource reuse.

Objective: To explore the association between daytime light exposure intensity and duration and objectively assessed sleep parameters among primary school students, so as to provide a scientific basis for understanding the impact of light exposure on children s sleep quality. Methods: From October 2023 to May 2024, a total of 741 primary school students in grades 2 to 3 were recruited from two primary schools in Chuzhou City, Anhui Province. Participants were required to wear an illuminance meter and an actigraph concurrently for 5 consecutive days to simultaneously collect 24 hour light exposure, sleep parameters, and physical activity data. Objective sleep parameters included total sleep time, sleep efficiency, sleep onset latency, wake after sleep onset, and sleep fragmentation index. Generalized linear model was applied to analyze the associations of average daytime (overall, morning, afternoon) light exposure intensity and exposure duration at different illuminance thresholds with various sleep parameters among primary school students. Results: The mean values of total sleep duration, sleep efficiency, sleep latency, wake time after sleep onset and sleep fragmentation index were (465.70±40.27)min, (83.22±5.16)%, (17.40±8.31)min, (75.97±26.81)min and (26.23±6.20), respectively. Generalized linear model showed that the average daytime light exposure intensity and duration of exposure ≥1 000 lx were positively correlated only with total sleep duration ( β =0.94, 8.51, both P <0.05). Morning light exposure intensity was associated with higher sleep efficiency ( β =0.11), longer total sleep duration ( β =0.80), shorter sleep latency ( β = -0.20 ), shorter wake time after sleep onset ( β =-0.57) and lower sleep fragmentation index ( β =-0.11) (all P <0.05). The duration of morning light exposure at ≥1 000 lx also improved childrens sleep efficiency, total sleep duration, sleep onset latency, wake time after sleep onset and sleep fragmentation index ( β =1.74, 13.91, -4.43, -11.04, -2.01, all P <0.05). Conclusions There is a significant association between morning light exposure intensity, exposure duration ≥1 000 lx, and multiple objective sleep parameters among primary school students. Increasing morning outdoor activities or high illuminance indoor light exposure could serve as feasible intervention strategies to improve the sleep of primary school students.

Objective To analyze the short-term prognosis value of middle ear risk index(MERI)in predicting the poor hearing recovery 6 months after transcanal endoscopic ear surgery(TEES)for children with cholesteatoma-type middle ear infection and the related factors affecting poor hearing recovery after TEES surgery.Methods A total of 70 children with cholesteatoma-type middle ear infection who underwent TEES were selected as the research subjects,and their outcomes were followed up for 6 months postoperatively.Pure tone audiometry was performed to collect the air-bone-gap(ABG)values of children,and they were divided into two groups according to ABG values:the group with good prognosis(ABG≤20 dBHL,n=49)and the group with poor prognosis(ABG>20 dBHL,n=21).The general information and the postoperative outcomes of MERI,ABG and quality of life improvement(Otitis Media-6,OM-6 score)of the two groups were compared.The independent risk factors for poor hearing recovery of TEES for children with cholesteatoma-type middle ear infection were analyzed by multivariate Logistic regression analysis.Receiver operating characteristic(ROC)curve was developed to study the predictive value of MERI in patients with cholestatomatous otitis media with TEES.Results Compared with the good prognosis group,the disease course of children in the poor prognosis group was longer,the scores of MERI,ABG and OM-6 before and after surgery were higher,the proportion of high-risk was higher(P＜0.05).The multivariate Logistic regression analysis showed that longer course of illness,higher MERI,higher preoperative ABG values and higher preoperative OM-6 scores were independent risk factors for poor hearing recovery of TEES for children with cholesteatoma-type middle ear infection.The ROC curve showed that the AUC(95%CI)of MERI predicting poor hearing recovery at 6 months post-TEES in cholesteatoma otitis media children was 0.828(95%CI:0.718-0.907),sensitivity was 81.0%and specificity was 75.5%.Conclusion MERI can be used as an effective tool to predict the short-term hearing recovery of children with cholestatoma otitis media undergoing tympanoplasty.The longer the course of disease,the higher the MERI.The higher the preoperative ABG,the higher the preoperative OM-6 score.The higher the risk of poor hearing recovery after TEES operation in children with cholestatoma otitis media,which needs clinical attention.

BackgroundSevere mental disorders are characterized by high recurrence rate， high disability rate， high rates of harmful incidents， and low treatment-seeking rate， with affected patients demonstrating increased frequencies of dangerous behaviors. Ningxia Hui Autonomous Region has implemented community management for patients with severe mental disorders across the region since 2004， while the current status and regional characteristics of the managed patients remain unclear. ObjectiveTo analyze the current status of management and treatment of patients with severe mental disorders in Ningxia Hui Autonomous Region， and to explore their regional distribution characteristics， so as to provide references for optimizing regional prevention and control strategies. MethodsPatients with severe mental disorders diagnosed and registered in the Severe Mental Disorder Management Information Platform of Ningxia Hui Autonomous Region from August 1， 2011 to December 31， 2021 were selected. Patients' basic information， management indicators， and treatment metrics were extracted from the platform， followed by descriptive statistical analysis of the corresponding data. ResultsAs of December 31， 2021， the permanent resident population of Ningxia Hui Autonomous Region was 6 946 540， with 29 787 registered patients with severe mental disorders. The majority of the patients were female （50.25%）， aged 18-59 years （79.01%）， with educational level of junior high school or below （84.63%）， married （52.87%）， farmers （56.01%）， and diagnosed with schizophrenia （55.91%）， while ethnic minority patients accounted for a relatively high proportion （31.35%）. In 2021， the reported prevalence rate of severe mental disorders in Ningxia Hui Autonomous Region was 0.43%， with standardized management and regular medication adherence rates at 90.39% and 66.34%， respectively. The standardized management rate in 8 counties/districts （36.36%） was lower than the average level of Ningxia Hui Autonomous Region， while 10 counties/districts （45.45%） showed below-average medication adherence rates， of which 6 counties/districts（60.00%） were located in the south-central region. ConclusionPatients with severe mental disorders in Ningxia Hui Autonomous Region are predominantly young and middle-aged adults with low level of education， and those in the central-southern region demonstrate lower medication adherence. ［Funded by Key Research and Development Program Project of Ningxia Hui Autonomous Region （number， 2023BEG02029）］

BACKGROUND: Currently, lung cancer is one of the malignant tumors with a high morbidity and mortality all over the world. However, the exact mechanisms underlying lung cancer progression remain unclear. The tumor necrosis factor receptor associated factor (TRAF) family members are cytoplasmic adaptor proteins, which function as both adaptor proteins and ubiquitin ligases to regulate diverse receptor signalings, leading to the activation of nuclear factor kappa-B (NF-κB), mitogen-activated protein kinase (MAPK) and interferon regulatory factor (IRF) signaling. The aim of this study was to investigate the expression of TRAFs in different tissues and cancer types, as well as its mRNA expression, protein expression, prognostic significance and functional enrichment analysis in non-small cell lung cancer (NSCLC), in order to provide new strategies for the diagnosis and treatment of NSCLC. METHODS: RNA sequencing data from the The Genotype-Tissue Expression database was used to analyze the expression patterns of TRAF family members in different human tissues. RNA sequencing data from the Cancer Cell Line Encyclopedia database was used to analyze the expression patterns of TRAF family members in different types of cancer cell lines. RNA sequencing data from the The Cancer Genome Atlas (TCGA) database was used to analyze the mRNA levels of TRAF family members across different types of human cancers. Immunohistochemistry (IHC) analyses from HPA database were used to analyze the TRAF protein levels in NSCLC [lung adenocarcinoma (LUAD) and lung squamous carcinoma (LUSC)]. Overall survival analysis was performed by Log-rank test using original data from Kaplan-Meier Plotter database to evaluate the correlation between TRAF expressions and prognosis. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses were performed on the TRAF family-related genes using RNA sequencing data from the TCGA database for NSCLC. The correlation between the expression levels of TRAF family members and the tumor immune microenvironment was analyzed using the ESTIMATE algorithm based on RNA sequencing data from the TCGA database. RESULTS: The TRAF family members exhibited significant tissue-specific expression heterogeneity. TRAF2, TRAF3, TRAF6 and TRAF7 were widely expressed in most tissues, while the expressions of TRAF1, TRAF4 and TRAF5 were restricted to specific tissues. The expressions of TRAF family members were highly specific among different types of cancer cell lines. In mRNA database of LUAD and LUSC, the expressions of TRAF2, TRAF4, TRAF5 and TRAF7 were significantly upregulated; while TRAF6 did the opposite; moveover, TRAF1 and TRAF3 only displayed a significant upregulation in LUAD and LUSC, respectively. Except for TRAF3, TRAF4 and TRAF7, other TRAF proteins displayed an obviously deeper IHC staining in LUAD and LUSC tissues compared with normal tissues. Additionally, patients with higher expression levels of TRAF2, TRAF4 and TRAF7 had shorter overall survival; while patients with higher expression levels of TRAF3, TRAF5 and TRAF6 had significantly longer overall survival; however, no significant difference had been observed between TRAF1 expression and the overall survival. TRAF family members differentially regulated multiple pathways, including NF-κB, immune response, cell adhesion and RNA splicing. The expression levels of TRAF family members were closely associated with immune cell infiltration and stromal cell content in the tumor immune microenvironment, with varying positive and negative correlations among different members. CONCLUSIONS TRAF family members exhibit highly specific expression differences across different tissues and cancer types. Most TRAF proteins exhibit upregulation at both mRNA and protein levels in NSCLC, whereas, only upregulated expressions of TRAF2, TRAF4 and TRAF7 predict worse prognosis. The TRAF family members regulate processes such as inflammation, immunity, adhesion and splicing, and influence the tumor immune microenvironment.
Humans ; Carcinoma, Non-Small-Cell Lung/pathology* ; Lung Neoplasms/mortality* ; Prognosis ; Gene Expression Regulation, Neoplastic ; Tumor Necrosis Factor Receptor-Associated Peptides and Proteins/metabolism*

Objective To analyze the short-term prognosis value of middle ear risk index(MERI)in predicting the poor hearing recovery 6 months after transcanal endoscopic ear surgery(TEES)for children with cholesteatoma-type middle ear infection and the related factors affecting poor hearing recovery after TEES surgery.Methods A total of 70 children with cholesteatoma-type middle ear infection who underwent TEES were selected as the research subjects,and their outcomes were followed up for 6 months postoperatively.Pure tone audiometry was performed to collect the air-bone-gap(ABG)values of children,and they were divided into two groups according to ABG values:the group with good prognosis(ABG≤20 dBHL,n=49)and the group with poor prognosis(ABG>20 dBHL,n=21).The general information and the postoperative outcomes of MERI,ABG and quality of life improvement(Otitis Media-6,OM-6 score)of the two groups were compared.The independent risk factors for poor hearing recovery of TEES for children with cholesteatoma-type middle ear infection were analyzed by multivariate Logistic regression analysis.Receiver operating characteristic(ROC)curve was developed to study the predictive value of MERI in patients with cholestatomatous otitis media with TEES.Results Compared with the good prognosis group,the disease course of children in the poor prognosis group was longer,the scores of MERI,ABG and OM-6 before and after surgery were higher,the proportion of high-risk was higher(P＜0.05).The multivariate Logistic regression analysis showed that longer course of illness,higher MERI,higher preoperative ABG values and higher preoperative OM-6 scores were independent risk factors for poor hearing recovery of TEES for children with cholesteatoma-type middle ear infection.The ROC curve showed that the AUC(95%CI)of MERI predicting poor hearing recovery at 6 months post-TEES in cholesteatoma otitis media children was 0.828(95%CI:0.718-0.907),sensitivity was 81.0%and specificity was 75.5%.Conclusion MERI can be used as an effective tool to predict the short-term hearing recovery of children with cholestatoma otitis media undergoing tympanoplasty.The longer the course of disease,the higher the MERI.The higher the preoperative ABG,the higher the preoperative OM-6 score.The higher the risk of poor hearing recovery after TEES operation in children with cholestatoma otitis media,which needs clinical attention.

Thoracic anesthesia with spontaneous respiration represents a form of precision anesthesia meticulously customized to individual patients.Considering the more stringent requirements this anesthesia approach imposes on the regulation of respiratory function,the writing group of the"Consensus of Experts on the Management of Thoracic Anesthesia with Spontaneous Respiration"has formulated elaborate guidelines regarding indications and contraindications,preoperative evaluation,anesthesia implementation,common complications,and treatment strategies.This was accomplished by referencing relevant domestic and international literature and integrating it with actual clinical requirements.The objective is to standardize the rational application of this anesthesia method.

Thoracic anesthesia with spontaneous respiration represents a form of precision anesthesia meticulously customized to individual patients.Considering the more stringent requirements this anesthesia approach imposes on the regulation of respiratory function,the writing group of the"Consensus of Experts on the Management of Thoracic Anesthesia with Spontaneous Respiration"has formulated elaborate guidelines regarding indications and contraindications,preoperative evaluation,anesthesia implementation,common complications,and treatment strategies.This was accomplished by referencing relevant domestic and international literature and integrating it with actual clinical requirements.The objective is to standardize the rational application of this anesthesia method.

Uncovering the risk factors of pulmonary hypertension and its mechanisms is crucial for the prevention and treatment of the disease.In the current study,we showed that experimental periodontitis,which was established by ligation of molars followed by orally smearing subgingival plaques from patients with periodontitis,exacerbated hypoxia-induced pulmonary hypertension in mice.Mechanistically,periodontitis dysregulated the pulmonary microbiota by promoting ectopic colonization and enrichment of oral bacteria in the lungs,contributing to pulmonary infiltration of interferon gamma positive(IFNγ+)T cells and aggravating the progression of pulmonary hypertension.In addition,we identified Prevotella zoogleoformans as the critical periodontitis-associated bacterium driving the exacerbation of pulmonary hypertension by periodontitis,and the exacerbation was potently ameliorated by both cervical lymph node excision and IFNγ neutralizing antibodies.Our study suggests a proof of concept that the combined prevention and treatment of periodontitis and pulmonary hypertension are necessary.

Objective:To explore the clinical and genetic characteristics of a fetus diagnosed with Congenital myasthenic syndrome type 16 (CMS16).Methods:A couple who had visited Tianjin Medical University General Hospital in February 2018 due to "adverse outcome of two pregnancies" was selected as the study subject. Clinical data was gathered. Peripheral blood and amniotic fluid samples were collected and subjected to whole exome sequencing (WES). Candidate variant was verified by Sanger sequencing. Low-depth whole-genome sequencing was carried out to detect copy number variation (CNV) in the fetus.Results:The couple′s first pregnancy had resulted in a miscarriage at 27 + 5 weeks, when ultrasound had revealed pleural effusion and polyhydramnios in the fetus. Their second pregnancy was terminated at 30 + 5 weeks due to fetal hand malformations, polyhydramnios and pleural fluid. Both couple had denied family history of genetic conditions. For their third pregnancy, no CNV abnormality was detected, whilst a compound heterozygous variants, including a maternally derived c. 3172C>T (p.R1058W) and paternal c. 1431delG (p.K477fs*89) in the SCN4A gene were detected. Based on the guidelines from the American College of Medical Genetics and Genomics, the c. 3172C>T (p.R1058W) was predicted as a likely pathogenic variant (PM1+ PM2_supporting+ PP3+ PP4), whilst the c. 1431delG (p.K477fs*89) was predicted as a pathogenic variant (PVS1+ PM2_supporting+ PP4). Conclusion:The c. 3172C>T (p.R1058W) and c. 1431delG (p.K477fs*89) compound heterozygous variants of the SCN4A gene probably underlay the CMS16 in the third fetus.