ObjectiveTo investigate the effect and molecular mechanism of exosomes derived from bone marrow mesenchymal stem cells（BMSC-exos） modified with Bushen Yisui capsule（BSYS）-containing serum on promoting the differentiation and maturation of OLN-93 oligodendrocytes by regulating miR-15b/Wnt signaling pathway. MethodsOLN-93 cells were divided into 5 groups， including the normal（NC） group， BMSC-exos group， BSYS-BMSC-exos group， BSYS-BMSC+LV-miR-15b-5p inhibitor-exos group， and BSYS-BMSC+LV-miR-15b-5p NC-exos group. DiR staining was used to observe the uptake of Exos by OLN-93 cells. The effective dosage of BSYS-BMSC-exos on OLN-93 cells was assessed by cell proliferation and activity assay（CCK-8）. Stable BMSCs lentiviral transfection strains were established to inhibit miR-15b-5p expression in both BMSCs and their exos， and transfection efficiency was verified by real-time fluorescent quantitative polymerase chain reaction（Real-time PCR） detection of miR-15b-5p. The expressions of 2′，3′-cyclic nucleotide 3′-phosphodiesterase（CNPase） and myelin proteolipid protein（PLP） in OLN-93 cells were detected by immunocytochemistry（ICC） and Western blot. The mRNA expressions of miR-15b-5p and Wnt3a in OLN-93 cells were detected by Real-time PCR， and the protein expression of Wnt3a was measured by Western blot. The expression levels of key molecules in the Wnt/β-catenin signaling pathway of OLN-93 cells， including glycogen synthase kinase（GSK）-3β， β-catenin， and T-cell specific transcription factor 4/transcription factor 7-like 2（TCF4/TCF7L2）， were measured by Real-time PCR and Western blot. ResultsDiR-labeled Exos were efficiently taken up by OLN-93 cells. The CCK-8 assay results indicated that 20 mg·L^-1 of BSYS-BMSC-exos exhibited the most significant effect in enhancing OLN-93 cell viability（P<0.01） and this dosage was selected for subsequent experiments. Following lentiviral transfection of BMSCs， Real-time PCR results revealed that miR-15b-5p was significantly suppressed in BMSCs（P<0.01）， and miR-15b-5p was also notably inhibited in BSYS-BMSC-exos（P<0.01）. ICC analysis further revealed an increase in the number of differentiated， mature CNPase and PLP-positive cells following BSYS-BMSC-exos treatment（P<0.01）. Western blot results demonstrated that the protein expression of CNPase and PLP was significantly enhanced with BSYS-BMSC-exos treatment（P<0.01）. Additionally， BSYS-BMSC-exos also increased the expression levels of miR-15b-5p and p-β-catenin proteins in OLN-93 cells， while decreased the mRNA and protein expressions of Wnt3a， as well as the mRNA expressions of β-catenin and TCF4/TCF7L2， and the protein expression level of p-GSK-3β（Ser9） was significantly reduced（P<0.05， P<0.01）. After the transfection of miR-15b-5p inhibitor into BSYS-BMSC-exos， the above effects were significantly diminished（P<0.05， P<0.01）. ConclusionBSYS-BMSC-exos facilitate the differentiation and maturation of OLN-93 cells， and its mechanism is related to the upregulation of miR-15b-5p in OLN-93 cells， which inhibits the expression of Wnt3a and thereby suppresses the Wnt signaling pathway.

ObjectiveTo explore the current situation and influencing factors of humanistic care experience among patients visiting traditional Chinese medicine （TCM） outpatient clinics in China， and to provide a basis for optimizing TCM-characterized services in both TCM and Western medicine hospitals. MethodsA multi-center cross-sectional study was conducted using convenience sampling to select 35 hospitals across 13 provinces in China （including 3 TCM hospitals and 32 TCM outpatient clinics in general hospitals）. A total of 3，430 patients were surveyed using the general information questionnaire and the Outpatient Humanistic Care Experience Questionnaire， with data collected via Questionnaire Star. Univariate analysis and multiple linear regression were employed to examine the impacts of patient characteristics， visit characteristics， hospital type （TCM hospital/general hospital）， and geographic region （eastern/central/western） on humanistic care experience. ResultsThe total score of humanistic care experience was 194 （188， 233）. Univariate analysis showed that gender， educational level， current residence， per capita monthly household income， location attribute of medical institutions， number of previous visits to this hospital， payment method of medical expenses， previous hospitalization history in this hospital， frequency of outpatient visits within the past 12 months， self-rated disease severity， familiarity with the outpatient procedures， implementation of the follow-up service provided by the hospital， satisfaction with follow-up services， the grade of the hospital visited， geographical region of the hospital visited， and the department visited had an impact on the humanistic care experience during outpatient visits （P<0.05）. Multivariate analysis demonstrated that educational level （β=0.609， P=0.011）， self-rated disease severity （β=-0.646， P=0.047）， familiarity with outpatient procedures （β=4.784， P<0.001）， satisfaction with follow-up services （β=6.365， P<0.001）， and the grade of the hospital visited （β=-5.487， P<0.001） affected the humanistic care experience in outpatient medical treatment， explaining 24.4% of the total variation. ConclusionHumanistic care experience in TCM outpatient clinics is influenced by multiple factors. It is recommended to optimize the medical treatment process， strengthen doctor-patient communication training， and establish a precise follow-up mechanism， with a focus on improving care perceptions among patients with lower education levels and those attending primary-level hospitals， to refine the TCM-characterized service system.

OBJECTIVES: To investigate the role of the calcium/calmodulin (CaM)-mediated activation of calcineurin (CN)-nuclear factor of activated T cells (NFAT) signaling pathway in mediating the regulatory effect of hyperforin (HY) on stress-induced depression-like disorder (DP) in mice. METHODS: C57BL/6J mice were randomly divided into control group, DP model group, and hyperforin treatment group (n=15). Behavioral changes of the mice were assessed using open field test (OFT), sucrose preference test (SPT), tail suspension test (TST), light/dark box test (LDB), and novel object suppression test (NSFT). Immunohistochemistry was used to detect tyrosine hydroxylase (TH) expression in the CA1 region of the hippocampus, and serum serotonin (5-HT) and norepinephrine (NA) levels were detected with ELISA. Western blotting was used to analyze the expressions of TNF-α, IL-1β, IL-2, and CN-NFAT pathway proteins. In cultured BV-2 microglial cells with lipopolysaccharide (LPS) stimulation, the effects of hyperforin and CN inhibitor (CNIS) on expressions of ionized calcium-binding adapter molecule 1 (IBA-1), 5-HT, NA, inflammatory cytokines and CN-NFAT pathway proteins were examined using immunofluorescence assay, ELISA or Western blotting. RESULTS: Compared with the control mice, the mice in DP group showed significantly reduced activity in OFT, decreased sucrose consumption in SPT, reduced shuttle crossing in LDB, and lowered food intake in NSFT with significantly increased immobility in TST. The mice with DP showed significantly decreased TH-positive neurons, lowered 5-HT and NA levels, and increased expressions of TNF-α, IL-1β, IL-2 and CaM-CN-NFAT pathway proteins. In cultured BV-2 cells, LPS stimulation strongly increased cellular IBA-1 expression, decreased the levels of neurotransmitters (5-HT and NA), and increased the levels of inflammatory cytokines and CN-NFAT signaling, and these changes were effectively reversed by treatment with hyperforin or CNIS. CONCLUSIONS Hyperforin improves stress-induced depression-like behaviors in mice and activated BV-2 cells by targeting the CN-NFAT signaling pathway.
Animals ; Mice, Inbred C57BL ; Mice ; Microglia/drug effects* ; Depression/etiology* ; Perylene/pharmacology* ; Calcineurin/metabolism* ; NFATC Transcription Factors/metabolism* ; Calcium Signaling/drug effects* ; Stress, Psychological ; Phloroglucinol/pharmacology* ; Signal Transduction ; Male ; Behavior, Animal/drug effects* ; Terpenes

Objective To evaluate the effect of different operation methods on the early swallowing function in the patients with laryngocarcinoma.Methods A total of 138 patients with the first time of open laryngectomy in this hospital from January 2021 to December 2022 were selected as the research subjects by convenience sampling method.The patients were divided into the vertical laryngeal resection group(vertical group,n=34),horizontal laryngeal partial resection(horizontal group,26 cases),suprachloroid laryngeal par-tial resection annular hyoid epiglottis fixation group(SCPL-CHEP group,n=26)and total laryngeal resection group(total laryngeal group,n=52)according to the operation methods.The Anderson Dysphagia Scale,Syd-ney Dysphagia Scale and modified swale drinking water test were used to evaluate the swallowing function on the first day of postoperative oral feeding in the patients.Results The total scores and scores of various di-mensions of the Chinese version of Anderson Dysphagia Scale,total scores and scores of various dimensions of Sydney Swallowing Scale and the results of the modified swale drinking water test had statistical differences a-mong the various groups(P＜0.01);the above indexes had statistical difference between the total laryngeal group and the other groups(P＜0.01),but the above indexes had no statistical difference between the two groups in the horizontal group,vertical group and SCPL-CHEP group(P＞0.05).Conclusion In the patients with laryngeal cancer undergoing open laryngectomy,the dysphagia is less severe during early eating after to-tal laryngectomy,which has little impact on life.

ObjectiveTo demonstrate the long-term effect of Yishi Therapy for primary tinnitus and analyze the factors affecting the effectiveness. MethodsWe conducted long-term follow-ups of primary tinnitus cases treated with Yishi Therapy during 1 January 2017 to 30 June 2023 by face-to-face， online and telephone between 1 July 2023 and 31 January 2024， to understand the final regression of primary tinnitus and lifestyle changes. We retrospectively analysed the basic data of patients before treatment， and statistically analysed the recovery rate and overall effective rate of tinnitus treated by the Yishi Therapy， and the factors affecting the effectiveness such as lifestyle， age， duration of disease， gender， tinnitus location， severity of tinnitus， accompanying symptoms and complications on the overall effective rate. ResultsA total of 496 cases of primary tinnitus completed follow-ups successfully， and the total Tinnitus Evaluation Questionnaire （TEQ） at follow-up was significantly lower than that before treatment （P＜0.001）. Among the 496 cases， 101 （20.36%） cases cured， 189 （38.10%） cases showed significant improvement， 116 （23.39%） cases were effective， 90 （18.15%） cases were ineffective， and the overall effect rate was 81.85% （406/496）. At follow-up， the score of lifestyle was higher than that before treatment （P＜0.001）. Grouped by the score of lifestyle at follow-up， the effective rates of better lifestyle group and poor lifestyle group were 97.53% （237/243） and 66.80% （169/253）， respectively， with statistical significance （P＜0.001）. Grouped by the mean value of the difference in lifestyle scores before and after treatment， the effective rates of the group with greater lifestyle adjustment and the group with smaller adjustment were 97.55% （239/245） and 66.53% （167/251）， respectively， and the differences were statistically significant （P＜0.001）. Binary logistic regression showed that the better lifestyle， the greater lifestyle adjustment and the severity of tinnitus were positively correlated with the total effective rate of tinnitus （P＜0.001）， while older and hearing loss were negatively correlated with the total effective rate of tinnitus （P＜0.05）. ConclusionYishi Therapy on primary tinnitus shows good long-term effect， lifestyle， severity of tinnitus， hearing loss or not， age are the relevant factors affecting the efficacy of tinnitus， lifestyle is the main factor affecting clinical efficacy.

Objective To understand the current status and influencing factors of patient perception for humanistic care in China hospitals,and to provide a basis for developing nursing humanistic care measures and improving the quality of nursing humanistic care services.Methods A total of 30,099 outpatients and inpatients from 107 hospitals in 30 provinces(autonomous regions and municipalities)from July to August 2022 as survey subjects.A general information questionnaire and the Relational Caring Questionnaire-Patient Form were used for a cross-sectional survey,and a single-factor analysis was used to analyze the influencing factors of patient relationship care.Results Finally,29 108 valid questionnaires were collected,and the effective questionnaire recovery rate was 96.7％.The patient evaluation of relationship care was(65.72±8.61)points.Single-factor analysis showed that gender,age,marital status,children's situation,education level,occupation,place of residence,average family income,medical insurance type,visiting department,and location of the visiting hospital,and whether or not surgery were influencing factors of patient relationship care(P＜0.05).Conclusion The evaluation score of caregiver-patient relationship care among Chinese hospital patients is above average,but there is still room for improvement in western and rural regions,seriously ill and outpatient patients,low-income and low-medical insurance reimbursement populations,and non-surgical patients.Medical institutions at all levels should optimize and improve nursing humanistic care services based on influencing factors,and further enhance patients'perception of nursing humanistic care.

Objective:To report the results of national surveillance on the distribution and antimicrobial resistance profile of clinical Gram-negative bacteria isolates from bloodstream infections in China in 2022.Methods:The clinical isolates of Gram-negative bacteria from blood cultures in member hospitals of national bloodstream infection Bacterial Resistant Investigation Collaborative System（BRICS）were collected during January 2022 to December 2022. Antibiotic susceptibility tests were conducted by agar dilution or broth dilution methods recommended by Clinical and Laboratory Standards Institute（CLSI）. WHONET 5.6 and SPSS 25.0 software were used to analyze the data.Results:During the study period，9 035 strains of Gram-negative bacteria were collected from 51 hospitals，of which 7 895（87.4%）were Enterobacteriaceae and 1 140（12.6%）were non-fermenting bacteria. The top 5 bacterial species were Escherichia coli（ n=4 510，49.9%）， Klebsiella pneumoniae（ n=2 340，25.9%）， Pseudomonas aeruginosa（ n=534，5.9%）， Acinetobacter baumannii complex（ n=405，4.5%）and Enterobacter cloacae（ n=327，3.6%）. The ESBLs-producing rates in Escherichia coli， Klebsiella pneumoniae and Proteus spp. were 47.1%（2 095/4 452），21.0%（427/2 033）and 41.1%（58/141），respectively. The prevalence of carbapenem-resistant Escherichia coli（CREC）and carbapenem-resistant Klebsiella pneumoniae（CRKP）were 1.3%（58/4 510）and 13.1%（307/2 340）；62.1%（36/58）and 9.8%（30/307）of CREC and CRKP were resistant to ceftazidime/avibactam combination，respectively. The prevalence of carbapenem-resistant Acinetobacter baumannii（CRAB）complex was 59.5%（241/405），while less than 5% of Acinetobacter baumannii complex was resistant to tigecycline and polymyxin B. The prevalence of carbapenem-resistant Pseudomonas aeruginosa（CRPA）was 18.4%（98/534）. There were differences in the composition ratio of Gram-negative bacteria in bloodstream infections and the prevalence of main Gram-negative bacteria resistance among different regions，with statistically significant differences in the prevalence of CRKP and CRPA（ χ2=20.489 and 20.252， P<0.001）. The prevalence of CREC，CRKP，CRPA，CRAB，ESBLs-producing Escherichia coli and Klebsiella pneumoniae were higher in provinicial hospitals than those in municipal hospitals（ χ2=11.953，81.183，10.404，5.915，12.415 and 6.459， P<0.01 or <0.05），while the prevalence of CRPA was higher in economically developed regions（per capita GDP ≥ 92 059 Yuan）than that in economically less-developed regions（per capita GDP <92 059 Yuan）（ χ2=6.240， P=0.012）. Conclusions:The proportion of Gram-negative bacteria in bloodstream infections shows an increasing trend，and Escherichia coli is ranked in the top，while the trend of CRKP decreases continuously with time. Decreasing trends are noted in ESBLs-producing Escherichia coli and Klebsiella pneumoniae. Low prevalence of carbapenem resistance in Escherichia coli and high prevalence in CRAB complex have been observed. The composition ratio and antibacterial spectrum of bloodstream infections in different regions of China are slightly different，and the proportion of main drug resistant bacteria in provincial hospitals is higher than those in municipal hospitals.

Outpatient humanistic care refered to providing a full process of caring medical services to outpatients. In order to standardize the human caring services for outpatients in medical institutions, promote the comprehensive service level of outpatient services, and improve the patient′s medical experience, Chinese Association for Life Care issued the group standard of Management Norms for Human caring of Outpatients in April 2023. This standard clarified the relevant terms and definitions of human caring for outpatients, specified the basic requirements for human caring, the humanistic quality and care responsibilities of outpatient staff, the outpatient care environment and facilities, the outpatient care process and measures, and quality management. It designed standardized and personalized full process care service norms, providing references for medical institutions at all levels to promote the development of human caring for outpatients.

AIM:To describe the dynamic characteristics of mitochondria in the development of mouse embryonic hematopoietic stem cells,and to explore the function of mitochondria in this process.METHODS:Single-cell transcrip-tomic data of continuous developmental hematopoietic stem cell-related populations were analyzed to describe the dynamic changes of genes related to mitochondrial synthesis and energy metabolism.To explore the dynamic changes in the number and activity of mitochondria during the development of hematopoietic stem cells,we detected the mitochondrial number and mitochondrial membrane potential of the cells in each stage of hematopoietic stem cell development by fluorescent probe staining combined with flow cytometry.We added small molecule inhibitors of mitochondrial synthesis and energy metabolism and used hematopoietic cell colony formation assay to detect the effect of mitochondrial function inhibition on the induction of hematopoietic products in vitro.RESULTS:(1)Single-cell transcriptome analysis showed that genes in-volved in mitochondrial synthesis and oxidative phosphorylation were significantly up-regulated in endothelial cell and type Ⅰ pre-hematopoietic stem cell compared with those involved in glycolysis,and these genes could significantly distinguish continuous dynamic populations.(2)The results of fluorescence staining and flow cytometry analysis showed that mito-chondrial number and mitochondrial membrane potential had an increasing trend during the continuous development of he-matopoietic stem cell,reaching the highest level in the precursor stage of type 2 pre-hematopoietic stem cell,and decreasing in the mature hematopoietic stem cell of fetal liver.(3)Compared with control group,inhibition of mitochondrial respirato-ry chain Ⅰ and Ⅴ significantly reduced the number of hematopoietic colonies(P<0.05).CONCLUSION:(1)Genes re-lated to mitochondrial synthesis and oxidative phosphorylation are highly expressed in hemogenic endothelial cells and type Ⅰpre-hematopoietic stem cells,and can be used to distinguish continuous developing populations.(2)The mitochondrial number and mitochondrial membrane potential increased continuously during hematopoietic stem cell development and reached the highest level in type 2 pre-hematopoietic stem cells.(3)Inhibition of mitochondrial respiratory chain Ⅰ and Ⅴ significantly reduced the production of hematopoietic products in vitro.

Objective:To explore the effects of swallowing rehabilitation training on swallowing function and quality of life in patients after laryngectomy.Methods:Randomized controlled trials on the effects of swallowing rehabilitation training on swallowing function and quality of life in postoperative laryngeal cancer patients were electronically retrieved from the Cochrane Library, PubMed, Embase, Web of Science, CINAHL, China National Knowledge Infrastructure, WanFang Data, VIP, and China Biology Medicine disc. The search period was from database establishment to February 1, 2024. The quality evaluation criteria for randomized controlled trials of Joanna Briggs Institute Evidence-Based Health Care Center (2016) was used to evaluate the included literature. RevMan 5.3 software was used for meta-analysis.Results:A total of ten articles were included, with 987 patients. Meta-analysis showed that swallowing rehabilitation training could improve the swallowing function ( P<0.01) and quality of life ( P<0.01) of patients after laryngectomy. Conclusions:Swallowing rehabilitation training can improve patients' swallowing function and quality of life after laryngectomy and is worth applying in clinical practice.