Objective To investigate the distribution and antimicrobial resistance profiles of clinically isolated Haemophilus influenzae and Moraxella catarrhalis in hospitals across China from 2015 to 2021,and provide evidence for rational use of antimicrobial agents.Methods Data of H.influenzae and M.catarrhalis strains isolated from 2015 to 2021 in CHINET program were collected for analysis,and antimicrobial susceptibility testing was performed by disc diffusion method or automated systems according to the uniform protocol of CHINET.The results were interpreted according to the CLSI breakpoints in 2022.Beta-lactamases was detected by using nitrocefin disk.Results From 2015 to 2021,a total of 43 642 strains of Haemophilus species were isolated,accounting for 2.91％of the total clinical isolates and 4.07％of Gram-negative bacteria in CHINET program.Among the 40 437 strains of H.influenzae,66.89％were isolated from children and 33.11％were isolated from adults.More than 90％of the H.influenzae strains were isolated from respiratory tract specimens.The prevalence of β-lactamase was 53.79％in H.influenzae strains.The H.influenzae strains isolated from children showed higher resistance rate than the strains isolated from adults.Overall,779 strains of H.influenzae did not produce β-lactamase but were resistant to ampicillin(BLNAR).Beta-lactamase-producing strains showed significantly higher resistance rates to these antimicrobial agents than the β-lactamase-nonproducing strains.Of the 16 191 M.catarrhalis strains,80.06％were isolated from children and 19.94％isolated from adults.M.catarrhalis strains were mostly susceptible to both amoxicillin-clavulanic acid and cefuroxime,evidenced by resistance rate lower than 2.0％.Conclusions The emergence of antibiotic-resistant H.influenzae due to β-lactamase production poses a challenge for clinical anti-infective treatment.Therefore,it is very important to implement antibiotic resistance surveillance for H.influenzae and guide rational antibiotic use.All local clinical microbiology laboratories should actively improve antibiotic susceptibility testing and strengthen antibiotic resistance surveillance for H.influenzae.

Oral squamous cell carcinoma(OSCC)is a malignant lesion originating from the oral mucosal squamous epithelium,account-ing for over 80％of oral and maxillofacial malignancies.Key etiological factors include tobacco,alcohol abuse,and betel quid chewing.In China,its incidence has shown an overall upward trend,posing a significant threat to public health.OSCC exhibits high local invasive-ness,making early diagnosis critical for improving prognosis.Its clinical management requires close multidisciplinary collaboration among oral and maxillofacial surgery,head and neck surgery,radiation oncology,medical oncology,reconstructive surgery,radiology,patholo-gy,and nutritional support teams.Given the increasing disease burden of OSCC and rapid development of multidisciplinary collaborative models,an expert panel has formulated this integrated management consensus based on evidence-based medicine and extensive deliber-ation.Centered on the'Prevention-Screening-Diagnosis-Treatment-Rehabilitation'framework,the consensus provides comprehensive guidance for the entire disease course of OSCC patients,aiming to standardize clinical practice.

This research investigated the contamination level,distribution of drug-resistant strains,and molecular epidemiologi-cal characteristics of Campylobacter,and further explored transmission pathways and prevention strategies.Cecum,chicken carcass,chicken product,and environmental samples,as well as swabs from workers'hands,were collected from a slaughterhouse in a large broiler group in the Jiaodong area between August 2023 and July 2024.Quantitative contamination assessment of Campylobacter in chicken carcasses and chicken products was performed.After microbial mass spectrometry identification,the representative strains of different links were selected for drug resistance testing and whole genome sequencing(WGS).On the basis of the sequencing results,the resistance genes,virulence genes,multilocus sequence typing(MLST),and phylogenetic characteristics of representative strains were analyzed.Homology comparisons were performed between isolates and strains from patients with diarrhea in the NCBI database.A total of 297 Campylobacter strains were isolated from 806 samples,and the overall detection rate was 36.85%.The detection rate of Campylobacter was highest in the evisceration process(47.33%),followed by the cutting process(35.64%).Overall,the Campylo-bacter detection rate first increased,then decreased,and subsequently increased.Drug sensitivity testing revealed that 90 isolates were resistant to nalidixic acid and ciprofloxacin,and 94.97%of isolates were resistant to tetracycline.WGS showed that both Campylo-bacter jejuni(C.jejuni)and Campylobacter coli(C.coli)carried many drug resistance and virulence genes.ST-14176 of C.jejuni was isolated for the first time herein.The predominant ST-8261 strain of C.jejuni and ST-860,ST-829,and ST-1586 strains of C.coli are known to cause human diarrhea.LOS expression genes associated with Guillain-Barré syndrome(GBS)were detected in both C.jejuni isolates from the slaughter chain and patients with GBS.Some strains exhibited close genetic relatedness to human-derived Campylo-bacter strains from the NCBI database.The detection rate of Campylobacter in the slaughterhouse first increased,then decreased,and subsequently increased,and the quantitative contamination level of each link was similar to the detection rate.Quantitative analysis of chicken carcasses/products revealed that the average bacterial load was highest in eviscerated carcasses(102.80 cfu/g),and the high-est amount of Campylobacter in chicken products reached 451.80 cfu/g.Abundant drug resistance genes and virulence genes were iden-tified,and the drug resistance genes were highly correlated with the drug resistance rate.Therefore,surveillance intensity and control measures for Campylobacter in slaughter processes should be strengthened.

Objective:To analyze the treatment-free remission (TFR) outcomes after discontinuation of tyrosine kinase inhibitor (TKI) in children with chronic myeloid leukemia (CML).Methods:In this retrospective cohort study, clinical data of 14 chronic phase CML children aged <18 years who had achieved stable deep molecular response (DMR) for ≥ 2 years after standardized treatment with TKI and had a strong desire to discontinue TKI at Henan Cancer Hospital from September 30, 2016 to January 30, 2022 were collected retrospectively. According to the different TFR outcomes after discontinuation of TKI, patients were divided into loss of major molecular response (MMR) group and without loss of MMR group, differences in clinical characteristics between the two groups of children were analyzed using Mann-Whitney U test and Fisher exact test. Results:Out of 14 children with TKI discontinuation, 7 were male and 7 were female. The age at diagnosis was 14.0 (4.8, 17.0) years, and the age at TKI discontinuation was 22.0 (12.5, 27.0) years. Among them, 8 children were treated with imatinib prior to TKI discontinuation and 6 children were treated with second-line substitution of the second-generation TKI nilotinib or dasatinib prior to TKI discontinuation. The follow-up time was 37.0 (27.8, 47.5) months, and 7 cases lost MMR at the time of discontinuation of 3.0 (2.0, 11.0) months. Eight children gained TFR at 6 months, 7 children gained TFR at 12 and 24 months. Amongst the 6 children who received second-generation TKI prior to TKI discontinuation, 2 children lost MMR at 3 and 11 months and 4 children gained TFR, among the 8 children who discontinued imatinib, 5 children lost MMR at the time 3.0 (2.0, 9.0) months and 3 children gained TFR. The age at diagnosis and TKI discontinuation, the time from TKI treatment to the acquisition of DMR, the duration of TKI treatment before TKI discontinuation, the duration of DMR before TKI discontinuation, and the number of children treated with second-generation TKI were not statistically different between the 7 children in the group that did not lose the MMR and the 7 children in the group that lost the MMR (all P>0.05) . All the 7 children with confirmed loss of MMR immediately restarted TKI therapy, and all regained DMR after 2.0 (2.0, 11.0) months of therapy. None of the children had disease progression. After TKI discontinued, only 1 child had mild bone pain, which could be relieved by oral antipyretic analgesic drugs. Conclusions:Children with CML who have achieved a durable stable DMR for≥2 years on TKI therapy can discontinue the TKI and obtain TFR. Both the longer duration of TKI therapy, the longer duration of DMR and the use of second-generation TKI therapy before TKI discontinuation, may allow more children with CML who are expecting TKI discontinuation to have access to TFR.

Buyang Huanwu Decoction(BYHWD), as one of the classic formulas in traditional Chinese medicine(TCM) for the treatment of cerebral ischemic stroke(CIS), has demonstrated definite effects in clinical practice. However, the material basis and mechanism of treatment have not been systematically elucidated. This study employed network pharmacology and molecular docking to analyze the potential targets and mechanisms of blood-and brain-penetrating active components of BYHWD in reducing cell apoptosis in CIS. Cell experiments were then carried out to validate the prediction results. In the experiments, five active components including hydroxysafflor yellow A( HSYA), tetramethylpyrazine( TMP), astragaloside Ⅳ( AS-Ⅳ), amygdalin( AMY), and paeoniflorin(PF) were selected to explore the pharmacological effects of BYHWD. HT22 cells were treated with BYHWD, and the cell counting kit-8(CCK-8) method was employed to examine the toxic and side effects of BYHWD. A cell model of oxygen-glucose deprivation/reoxygenation( OGD/R) was constructed, with apoptosis and pyroptosis as the main screening indicators. The levels of lactate dehydrogenase(LDH) and glutathione(GSH) were measured to assess the cell membrane integrity. Flow cytometry was employed to detect apoptosis, and the activities of caspase-3 and caspase-1 were measured to clarify the status of apoptosis and pyroptosis. ELISA was employed to determine the levels of interleukin(IL)-1β and IL-18 to confirm pyroptosis. HSYA and AMY were identified in this study as the active components regulating apoptosis and pyroptosis. TUNEL was employed to detect the apoptosis rate, and Western blot was employed to determine the expression levels of apoptosis-related proteins B-cell lymphoma-2(Bcl-2), Bcl-2-associated X protein(Bax), and caspase-3, which confirmed that the anti-apoptotic effect of the combined component group was superior to that of the single component groups. The molecular docking results revealed strong binding affinity of HSYA and AMY with SDF-1α and CXCR4.AMD3100, a selective antagonist of CXCR4, was then used for intervention. The results of Western blot showed alterations in the expression levels of apoptosis-associated proteins, SDF-1α, and CXCR4. In conclusion, HSYA and AMY influence cellular apoptosis by modulating the SDF-1α/CXCR4 signaling cascade.
Drugs, Chinese Herbal/chemistry* ; Apoptosis/drug effects* ; Animals ; Neurons/cytology* ; Mice ; Molecular Docking Simulation ; Cell Line ; Glucose/metabolism* ; Humans ; Neuroprotective Agents/pharmacology*

This study aims to explore the effects and action mechanisms of the active ingredients in Buyang Huanwu Decoction(BYHWD), namely tetramethylpyrazine(TMP) and hydroxy-safflor yellow A(HSYA), on oxygen-glucose deprivation/reglucose-reoxygenation(OGD/R)-induced inflammation and oxidative stress of microglia(MG). Network pharmacology was used to screen the effective monomer ingredients of BYHWD and determine the safe concentration range for each component. Inflammation and oxidative stress models were established to further screen the best ingredient combination and optimal concentration ratio with the most effective anti-inflammatory and antioxidant effects. OGD/R BV2 cell models were constructed, and BV2 cells in the logarithmic growth phase were divided into a normal group, a model group, an HSYA group, a TMP group, and an HSYA + TMP group. Enzyme-linked immunosorbent assay(ELISA) was used to detect the levels of inflammatory cytokines such as interleukin-1β(IL-1β), tumor necrosis factor-α(TNF-α), and interleukin-6(IL-6). Oxidative stress markers, including superoxide dismutase(SOD), nitric oxide(NO), and malondialdehyde(MDA), were also measured. Western blot was used to analyze the protein expression of both inflammation-related pathway [Toll-like receptor 4(TLR4)/nuclear factor-kappa B(NF-κB)] and oxidative stress-related pathway [nuclear factor erythroid 2-related factor 2(Nrf2)/heme oxygenase-1(HO-1)]. Immunofluorescence was used to assess the expression of proteins such as inducible nitric oxide synthase(iNOS) and arginase-1(Arg-1). The most effective ingredients for anti-inflammatory and antioxidant effects in BYHWD were TMP and HSYA. Compared to the normal group, the model group showed significantly increased levels of IL-1β, TNF-α, IL-6, NO, and MDA, along with significantly higher protein expression of NF-κB, TLR4, Nrf2, and HO-1 and significantly lower SOD levels. The differences between the two groups were statistically significant. Compared to the model group, both the HSYA group and the TMP group showed significantly reduced levels of IL-1β, TNF-α, IL-6, NO, and MDA, lower expression of NF-κB and TLR4 proteins, higher levels of SOD, and significantly increased protein expression of Nrf2 and HO-1. Additionally, the expression of the M1-type MG marker iNOS was significantly reduced, while the expression of the M2-type MG marker Arg-1 was significantly increased. The results of the HSYA group and the TMP group had statistically significant differences from those of the model group. Compared to the HSYA group and the TMP group, the HSYA + TMP group showed further significant reductions in IL-1β, TNF-α, IL-6, NO, and MDA levels, along with significant reductions in NF-κB and TLR4 protein expression, an increase in SOD levels, and elevated Nrf2 and HO-1 protein expression. Additionally, the expression of the M1-type MG marker iNOS was reduced, while the M2-type MG marker Arg-1 expression increased significantly in the HSYA + TMP group compared to the TMP or HSYA group. The differences in the results were statistically significant between the HSYA + TMP group and the TMP or HSYA group. The findings indicated that the combined use of HSYA and TMP, the active ingredients of BYHWD, can effectively inhibit OGD/R-induced inflammation and oxidative stress of MG, showing superior effects compared to the individual use of either component.
Oxidative Stress/drug effects* ; Drugs, Chinese Herbal/pharmacology* ; Animals ; Mice ; Glucose/metabolism* ; Cell Line ; Inflammation/genetics* ; Oxygen/metabolism* ; Pyrazines/pharmacology* ; Microglia/metabolism* ; NF-E2-Related Factor 2/immunology* ; NF-kappa B/immunology* ; Toll-Like Receptor 4/immunology* ; Anti-Inflammatory Agents/pharmacology* ; Humans

OBJECTIVE: To evaluate the dynamic changes of glucocorticoid (GC) dose and the feasibility of GC discontinuation in rheumatoid arthritis (RA) patients under the background of Chinese medicine (CM). METHODS: This multicenter retrospective cohort study included 1,196 RA patients enrolled in the China Rheumatoid Arthritis Registry of Patients with Chinese Medicine (CERTAIN) from September 1, 2019 to December 4, 2023, who initiated GC therapy. Participants were divided into the Western medicine (WM) and integrative medicine (IM, combination of CM and WM) groups based on medication regimen. Follow-up was performed at least every 3 months to assess dynamic changes in GC dose. Changes in GC dose were analyzed by generalized estimator equation, the probability of GC discontinuation was assessed using Kaplan-Meier curve, and predictors of GC discontinuation were analyzed by Cox regression. Patients with <12 months of follow-up were excluded for the sensitivity analysis. RESULTS: Among 1,196 patients (85.4% female; median age 56.4 years), 880 (73.6%) received IM. Over a median 12-month follow-up, 34.3% (410 cases) discontinued GC, with significantly higher rates in the IM group (40.8% vs. 16.1% in WM; P<0.05). GC dose declined progressively, with IM patients demonstrating faster reductions (median 3.75 mg vs. 5.00 mg in WM at 12 months; P<0.05). Multivariate Cox analysis identified age <60 years [P<0.001, hazard ratios (HR)=2.142, 95% confidence interval (CI): 1.523-3.012], IM therapy (P=0.001, HR=2.175, 95% CI: 1.369-3.456), baseline GC dose ⩽7.5 mg (P=0.003, HR=1.637, 95% CI: 1.177-2.275), and absence of non-steroidal anti-inflammatory drugs use (P=0.001, HR=2.546, 95% CI: 1.432-4.527) as significant predictors of GC discontinuation. Sensitivity analysis (545 cases) confirmed these findings. CONCLUSIONS RA patients receiving CM face difficulties in following guideline-recommended GC discontinuation protocols. IM can promote GC discontinuation and is a promising strategy to reduce GC dependency in RA management. (Trial registration: ClinicalTrials.gov, No. NCT05219214).
Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Arthritis, Rheumatoid/drug therapy* ; Glucocorticoids/therapeutic use* ; Medicine, Chinese Traditional ; Retrospective Studies

Objective:To investigate whether exosome-derived microRNA (miR)-877-5p can affect the malignant biological behaviors of glioma cells by regulating transmembrane 4 superfamily member 1 (TM4SF1).Methods:(1) Tumor tissues and corresponding adjacent tissues from 42 patients with glioma who underwent surgical resection in Department of Neurosurgery, the First Affiliated Hospital of Nanyang Medical College from September 2024 to February 2025 were collected. The miR-877-5p and TM4SF1 mRNA expressions in tumor tissues and corresponding adjacent tissues were detected by quantitative reverse transcription polymerase chain reaction (qRT-PCR), and the correlation between miR-877-5p and TM4SF1 mRNA expressions in tumor tissues was analyzed by Pearson correlation. (2) HEB, U87, LN229, and U251 cells at the logarithmic growth phase were cultured and their miR-877-5p and TM4SF1 mRNA expressions were detected by qRT-PCR. Exosomes from U87, LN229, and U251 cells were isolated, and their morphology was observed under a transmission electron microscope; protein expressions of CD9, CD63, tumor susceptibility gene 101 (TSG101), and Calnexin in exosomes were detected by Western blotting. The miR-877-5p expression in exosomes of U87, LN229, and U251 cells was detected by qRT-PCR. The diameter of exosomes from LN229 cells was measured using a Malvern Zetasizer particle size and zeta potential analyzer, and the uptake efficiency of exosomes in LN229 cells was detected by flow cytometry. LN229 cells were divided into a normal control group, a miR-877-5p negative control group, a miR-877-5p mimic group, a miR-877-5p mimic+pcDNA empty vector group, and a miR-877-5p mimic+pcDNA-TM4SF1 group; except for the normal control group, the other groups were transfected with corresponding plasmids through exosomes and cultured for 24 hours; and then, miR-877-5p mRNA expression was detected by qRT-PCR; cell viability was detected by CCK-8 assay, cell apoptosis was detected by flow cytometry, cell invasion was detected by Transwell assay, and TM4SF1, Cyclin D1, Bcl-2 associated X protein (Bax), and matrix metalloproteinase 2 (MMP2) protein expressions and expressions of TM4SF1 downstream pathway proteins phosphorylated protein kinase B (p-AKT) and β-catenin were detected by Western blotting. The targeting relation between miR-877-5p and TM4SF1 was validated using a dual-luciferase reporter assay. A U251 cell experiment was performed for universal verification: U251 cells were divided into a normal control group, a miR-877-5p negative control group, a miR-877-5p mimic group, a miR-877-5p mimic+pcDNA empty vector group, and a miR-877-5p mimic+pcDNA-TM4SF1 group; cell apoptosis was detected by flow cytometry, and TM4SF1 protein expression was detected by Western blotting. (3) Eighteen male BALB/c nude mice were randomly divided into a control group, a miR-877-5p mimic group, and a miR-877-5p mimic+pcDNA-TM4SF1 group, with 6 mice in each group; 100 μL LN229 cell suspension, LN229 cell suspension transfected with miR-877-5p mimic, and LN229 cell suspension transfected with miR-877-5p mimic and pcDNA-TM4SF1 were, respectively, subcutaneously injected into the lateral abdomen of nude mice in these 3 groups. After 28 days of feeding, the mass and volume of the transplanted tumors were measured, and the TM4SF1, Cyclin D1, Bax, and MMP2 protein expressions in the transplanted tumors were detected by Western blotting. Results:(1) Compared with that in the corresponding adjacent tissues, miR-877-5p mRNA expression in the tumor tissues was significantly decreased and TM4SF1 mRNA expression was statistically increased ( P<0.05). Correlation analysis showed that the miR-877-5p and TM4SF1 mRNA expressions in the tumor tissues were negatively correlated ( r=-0.966, P<0.001). (2) Compared with those in the HEB cells, statistically decreased miR-877-5p mRNA expression and increased TM4SF1 mRNA expression in U87, LN229, and U251 cells were noted ( P<0.05). Under the transmission electron microscope, the exosomes in glioma cells were all biconcave disc-shaped and had a complete lipid bilayer membrane structure. Western blotting indicated positive CD9, CD63, and TSG101 protein expressions and negative Calnexin protein expression in the exosomes of glioma cells. Flow cytometry results indicated a relatively high uptake efficiency of exosomes in LN229 cells. Compared with that in the U87 and U251 cells, the miR-877-5p mRNA expression in exosomes of LN229 cells was significantly decreased ( P<0.05). The diameter of exosomes in LN229 cells was 80-150 nm. Compared with the miR-877-5p negative control group, the miR-877-5p mimic group had an increased miR-877-5p mRNA expression, decreased cell survival rate (negative control group: [95.43±0.23]%; miR-877-5p mimic group: [51.24±5.67]%), increased cell apoptosis rate ([3.34±0.22]% vs. [35.24±4.17]%), reduced number of invasive cells ([127.33±13.63] cells per high-power field vs.[59.67±6.87] cells per high-power field), downregulated TM4SF1, Cyclin D1 and MMP2 protein expressions, upregulated Bax protein expression, and decreased p-AKT and β-catenin protein expressions, with significant differences ( P<0.05). Compared with the miR-877-5p mimics+pcDNA empty vector group, the miR-877-5p mimic+pcDNA-TM4SF1 group had a decreased miR-877-5p expression, increased cell survival rate (miR-877-5p mimics+pcDNA empty vector group: [56.27±5.24]%; miR-877-5p mimic+pcDNA-TM4SF1 group[75.31±8.13]%), decreased cell apoptosis rate ([36.27±4.42]% vs. [15.37±1.73]%), increased number of invasive cells ([62.67±6.14] cells per high-power field vs. [95.50±10.58] cells per high-power field), upregulated TM4SF1, Cyclin D1 and MMP2 protein expressions, decreased Bax protein expression, and upregulated p-AKT and β-catenin protein expressions, with significant differences ( P<0.05). Dual-luciferase assay results showed that in the plasmids carrying wild-type TM4SF1 sequence, the luciferase activity in the miR-877-5p mimics group was significantly lower than that in the miR-877-5p negative control group ( P<0.05); in the plasmids carrying the mutant TM4SF1 sequence, no significant change in the luciferase activity was noted between the miR-877-5p negative control group and miR-877-5p mimic group ( P>0.05). Universal verification results: in U251 cells, compared with the miR-877-5p negative control group, the miR-877-5p mimic group had a significantly increased cell apoptosis rate and a statistically decreased TM4SF1 protein expression ( P<0.05); compared with the miR-877-5p mimic+pcDNA empty vector group, the miR-877-5p mimic+pcDNA-TM4SF1 group showed significantly decreased apoptosis rate and statistically increased TM4SF1 protein expression ( P<0.05). (3) Compared with the normal control group, the miR-877-5p mimic group had statistically reduced tumor mass and volume, significantly decreased TM4SF1, Cyclin D1 and MMP2 protein expressions, and significantly increased Bax protein expression ( P<0.05). Compared with the miR-877-5p mimic group, the miR-877-5p mimic+pcDNA-TM4SF1 group had significantly increased tumor mass and volume, statistically increased TM4SF1, Cyclin D1 and MMP2 protein expressions, and statistically decreased Bax protein expression ( P<0.05). Conclusion:Exosome-derived miR-877-5p may inhibit the proliferative and invasive capacities of glioma cells and promote cell apoptosis by targetedly inhibiting the TM4SF1 expression, thereby exerting an anti-tumor effect.

Objective To analyze the influence path of medical students'professional identity on positive defensive medical behaviors and examine the mediating effect of medical risk perception between the two,aiming to provide theoretical support and practical reference for medical education to actively adapt to the demands of the medical and health system reform and promote the scientific guidance of positive defensive medical behaviors.Methods ln 2020,a questionnaire survey was conducted among medical students from four medical colleges in Heilongjiang Province using cluster sampling method,and data analysis was performed using SPSS 27.0 software.Results Medical students'professional identity showed a significant positive correlation with their positive defensive medical behavior(β=0.225,P＜0.001),with medical risk perception acting as a partial mediator(indirect effect accounting for 19%of the total effect).Professional identity(β=0.183,P＜0.001)and medical risk perception(β=0.195,P＜0.001)exerted direct positive effects on positive defensive medical behavior.Conclusion Medical students'professional identity drives them to adopt positive defensive medical behaviors by strengthening their sense of responsibility ethics and risk cognition ability,and medical risk perception is a key mediating variable in this process.Educational management should integrate policy-oriented clinical decision-making training and balance medical safety and efficiency through the collaborative mechanism of"professional identity-risk perception-behavioral practice".