BackgroundObstructive sleep apnea （OSA） represents a prevalent sleep disordered breathing condition characterized by complex pathophysiology. The arousal threshold （ArTH）， a core non-anatomical contributor to OSA pathogenesis， is intimately tied to the disease severity and clinical phenotypes. To date， research regarding factors associated with ArTH has yielded inconsistent findings， and ArTH profiles and disparities across populations with different genetic backgrounds are not fully elucidated. ObjectiveTo explore the differences of ArTH in OSA patients with different genetic backgrounds and analyze the key factors affecting ArTH， thereby providing evidence for understanding OSA pathophysiology and formulating targeted treatment regimens. MethodsA total of 285 patients who met the diagnostic criteria for OSA， as defined by the Multidisciplinary Diagnosis and Treatment Guidelines for Adult Obstructive Sleep Apnea， were retrospectively enrolled in this study. All participants underwent overnight polysomnography （PSG） at the Sleep Medicine Center of Inner Mongolia Mental Health Center from December 2022 to May 2024. Based on the study design， the cohort was stratified into two distinct genetic background subgroups （group A and group B）. Demographic and clinical characteristics， the Epworth Sleepiness Scale （ESS） score， and overnight PSG data were collected. The apnea hypopnea index （AHI）， the lowest pulse oxygen saturation （LSpO₂）， and fraction of hypopneas （F_Hypopneas） were utilized as surrogate indicators to estimate ArTH in OSA patients. The influencing factors of low ArTH were tested by binary Logistic regression analysis. ResultsAmong the 285 OSA patients， there were 227 cases （79.65%） in group A and 58 cases （20.35%） in group B. Comparisons between the two genetic background subgroups revealed no statistically significant differences in the proportion of low ArTH， ESS score， PSG parameters， and the three markers for low ArTH （AHI<30 events/h， LSpO₂>82.5%， F_Hypopneas>58.3%） （P>0.05）. Binary Logistic regression analysis identified sex （OR=2.421， 95% CI： 1.070–5.478）， BMI （OR=0.847， 95% CI： 0.770–0.932）， N1 sleep duration （OR=0.974， 95% CI： 0.963–0.985）， and hypertension （OR=0.348， 95% CI： 0.143–0.848） as independent factors associated with low ArTH in the total cohort. Stratified analysis by genetic backgrounds revealed that sex（OR=3.799， 95% CI： 1.389–10.392）， BMI（OR=0.819， 95% CI：0.723–0.929）， and N1 sleep duration（OR=0.973， 95% CI： 0.961–0.986） were independent factors of low ArTH in group A. In contrast， only N1 sleep duration （OR=0.951， 95% CI： 0.911–0.993） remained a significant factor in group B. ConclusionArthur may have conservative characteristics in patients with OSA with different genetic backgrounds， but the pathophysiological mechanism of OSA may have population heterogeneity. ［Funded by Inner Mongolia Autonomous Region Natural Science Fundation Project （number， 2024QN08050）； Intra-institutional Scientific Research Project of Inner Mongolia Mental Health Center （number， 2022QNWN0010）］

Objective:To evaluate the application of metagenomic next-generation sequencing（mNGS）in the identification of non-tuberculous mycobacteria（NTM）.Methods:A retrospective analysis was conducted on mNGS results of 358 bronchoalveolar lavage fluid（BALF）samples positive for NTM collected at the First Affiliated Hospital of Zhejiang University School of Medicine from February 2021 to January 2024. The analysis included the distribution of NTM species，the detection of mixed pathogens，and the performance of conventional mycobacterial detection methods.Results:The results showed that 362 strains of 15 NTM species were identified from 350 specimens，8 specimens were not precise to the species level. The most frequently detected species were Mycobacterium intracellulare（37.3%，135/362）， Mycobacterium abscessus（26.8%，97/362），followed by Mycobacterium avium（11.0%，40/362）， Mycobacterium kansasii（8.0%，29/362）and Mycobacterium chelonae（7.7%，28/362）. Single NTM species were detected in 339 specimens，while two or three NTM species were simultaneously detected in 11 specimens（3.1%，11/358）. Non-NTM microorganisms co-infected were detected in 53.4%（191/358）of NTM-positive BALF samples，including common pathogens such as Pseudomonas aeruginosa， Staphylococcus aureus，and Aspergillus fumigatus；and difficult-to-identify pathogens such as Legionella pneumophila and Talaromyces marneffei. In NTM-positive patients detected by mNGS，the results supported the diagnosis of NTM infection in 298 cases（298/358，83.2%）and 105 cases（105/358，29.3%）initiated anti-NTM treatment accordingly；while in 60 cases（60/358，16.8%）the positive results were considered as colonization or unrelated to clinical infection. For samples tested with acid-fast staining，mycobacterial liquid culture，and DNA microarray，the positivity rates for NTM were 31.5%（73/232），48.7%（57/117），and 43.0%（46/107），respectively. Conclusions:mNGS demonstrates advantages in identification of NTM. However，the test may detect multiple microorganisms，in that case，the interpretation with clinical and radiological results is requried to determine the main pathogens.

OBJECTIVE T o compare the biological characteristics,drug resistance and pathogenicity between two strains of mucoid Pseudomonas aeruginosa and the standard strain PAO1.METHODS The strains were identified,and biofilms were detected by 96-well plates method.The bacterial drug resistance was detected by fully automatic drug susceptibility analysis system,the expression levels of RNA of virulence factors were detected by real-time fluorescent quantitative polymerase chain reaction(RT-PCR);the models of rats with pneumonia infection were established through liquid aerosol lung delivery method,the survival status of the rats was observed,and the lev-els of cytokines in bronchoalveolar lavage fluid(BALF)were detected.RESULTS NY4593,NY4605 and PAO1 strains were successfully isolated and identified.NY4593 and NY4605 showed high-yield biofilms,while PAO1 showed low-yield biofilms.The drug resistance rates of NY4593 and NY4605 were remarkably higher than those of the PAO1.The expression levels of exoT and exoY gene RNA of the NY4593 and NY4605 strains were higher than those of the PAO1 strains(P＜0.05);the expression level of exoS gene RNA of the NY4605 was lower than that of the PAO1(P＜0.05).Under the same infection dose,the PAO1 showed more powerful pathogenicity,and the secretion volumes of inflammatory factors interleukin-6(IL-6),interleuki-1β(IL-1β)and interleukin-17A(IL-17A)of the PAO1 were(2858.00±150.30)pg/ml,(7821.00±761.20)pg/ml and(1079.00±225.40)pg/ml respectively,remarkably higher than those of the NY4593 and NY4605(P＜0.05).CONCLUSION The clini-cal mucoid NY4593 and NY4605 remarkably differ from PAO1 in biology and pathogenicity.The study may facilitate deep understanding of the mechanisms of PA infection and provide guidance for clinical treatment,prevention and control.

During tumor progression, the mechanical properties of the tumor microenvironment play a pivotal regulatory role. As core mechanical indicators, cellular stiffness and extracellular matrix stiffness profoundly influence tumor development through multiple pathways, including cytoskeletal remodeling, activation of signaling pathways, and metabolic regulation. Studies have demonstrated that the tissue stiffness of various solid tumors is significantly higher than that of corresponding normal tissues, while their cellular stiffness exhibits the opposite trend. This mechanical characteristic is also observed in oral squamous cell carcinoma and exerts crucial regulatory effects during tumor progression. This review systematically summarizes the molecular composition and regulatory mechanisms underlying the stiffness of tumor cells and extracellular matrix (ECM). Mainstream stiffness detection technologies such as atomic force microscopy, microfluidic deformation, and real-time deformability cytometry are outlined, with particular emphasis on their applications and limitations in oncology research. This review comprehensively analyzes how mechanical properties regulate key processes in tumor progression, including growth, proliferation, invasion, metastasis, angiogenesis, lymphangiogenesis, drug resistance, and immune escape. This review synthesizes biomechanics-based therapeutic strategies, including: ① targeting the regulation of tumor cell stiffness through cytoskeletal modulators and cholesterol-depleting agents to enhance immune responses; ② reducing ECM stiffness by matrix remodeling enzyme inhibitors, ECM component modulators, or receptor antagonists to improve drug delivery efficiency, and combining with immunotherapy or photothermal therapy for enhanced therapeutic effects; ③ enhancing the mechanical adaptability and anti-tumor activity of immune cells through pharmacological or genetic approaches. This review establishes a robust conceptual framework for developing novel anti-tumor therapeutic strategies and provides insights for future clinical management of oral squamous cell carcinoma.

ObjectiveTo investigate the effects of different concentrations of Shaoyaotang-containing serum on lipopolysaccharide （LPS）-induced inflammation of human colorectal adenocarcinoma （Caco-2） cells by inhibiting apoptosis via activating the tumor necrosis factor （TNF） receptor superfamily member 6 （Fas）/Fas ligand （FasL） pathway. MethodsCaco-2 cells were allocated into blank， model （LPS， 10 mg·L^-1）， Shaoyaotang-containing serum （5%， 10%， 15%， 20%）， and Fas inhibitor （KR-33493， 20 mmol·L^-1） groups. Except the blank group， the other groups were stimulated with 10 mg·L^-1 LPS for 24 h for the modeling of inflammation. After successful modeling， the blank， Fas inhibitor， and model groups were treated with blank serum， and the Shaoyaotang-containing serum groups were treated with the serum samples at corresponding concentrations for 24 h. The Fas inhibitor group was subjected to KR-33493 pretreatment for 1 h. Cell proliferation and viability were examined by the cell-counting kit-8 （CCK-8） method. The levels of interleukin （IL）-6， IL-1β， and TNF-α were measured by enzyme-linked immunosorbent assay. Apoptosis was detected by flow cytometry. The protein and mRNA levels of Fas， FasL， cysteinyl aspartate-specific proteinase （Caspase）-3， Caspase-9， B-cell lymphoma 2 （Bcl-2）， and Bcl-2-associated X protein （Bax） were determined by Western blot and Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）， respectively. ResultsCompared with the blank group， the model group presented a decrease in cell survival rate （P<0.01）. Compared with that in the model group， the cell survival rate showed no significant change in the 5% Shaoyaotang-containing serum group but increased in the 10%， 15%， and 20% Shaoyaotang-containing serum groups （P<0.01）. Since there was no statistical difference between the 5% Shaoyaotang-containing serum group and the model group， 10%， 15%， and 20% Shaoyaotang-containing sera were selected for the follow-up study. Compared with the blank group， the model group showed risen levels of IL-6， IL-1β， and TNF-α （P<0.01）， an increased apoptosis rate （P<0.01）， up-regulated protein and mRNA levels of Fas， FasL， Caspase-3， Caspase-9， and Bax （P<0.01）， and down-regulated protein and mRNA levels of Bcl-2 （P<0.01）. Compared with the model group， the Fas inhibitor group and the 10%， 15%， and 20% Shaoyaotang-containing serum groups showed declined levels of IL-6， IL-1β， and TNF-α （P<0.01）， decreased apoptosis rates （P<0.01）， down-regulated protein and mRNA levels of Fas， FasL， Caspase-3， Caspase-9， and Bax （P<0.05， P<0.01）， and up-regulated protein and mRNA levels of Bcl-2 （P<0.05， P<0.01）. In addition， the 15% and 20% Shaoyaotang-containing serum groups had lower levels of IL-6， IL-1β， and TNF-α （P<0.05， P<0.01）， lower apoptosis rates （P<0.05， P<0.01）， lower protein and mRNA levels of Fas， FasL， Caspase-3， Caspase-9， and Bax （P<0.05， P<0.01）， and higher protein and mRNA levels of Bcl-2 （P<0.05， P<0.01） than the 10% Shaoyaotang-containing serum group. ConclusionThe Shaoyaotang-containing serum can reduce the content of inflammatory factors in Caco-2 cells， down-regulate the protein and mRNA levels of Fas， FasL， Caspase-3， Caspase-9， and Bax， and up-regulate the protein and mRNA levels of Bcl-2 under the intervention of LPS by regulating the Fas/FasL pathway and inhibiting the apoptosis of intestinal epithelial cells in ulcerative colitis.

ObjectiveTo investigate the protective effect and mechanism of Shaoyaotang （SYD） on the lipopolysaccharide （LPS）-induced damage of tight junction proteins in the human colorectal adenocarcinoma （Caco-2） cell model of inflammation via the Ras homolog gene family member A （RhoA）/Rho-associated coiled-coil forming protein kinase （ROCK） pathway. MethodsCaco-2 cells were grouped as follows： Blank， model （LPS， 10 mg·L^-1）， SYD-containing serum （10%， 15%， and 20%）， and inhibitor （Fasudil， 25 μmol·L^-1）. After 24 hours of intervention， the cell viability in each group was examined by the cell-counting kit 8 （CCK-8） method. Enzyme-linked immunosorbent assay was employed to determine the levels of endothelin-1 （ET-1）， tumor necrosis factor-α （TNF-α）， interleukin-1β （IL-1β）， and interleukin-6 （IL-6）. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot were employed to determine the mRNA and protein levels， respectively， of RhoA， ROCK2， claudin-5， and zonula occludens-1 （ZO-1） in cells of each group. ResultsCompared with the blank group， the model group showcased a marked reduction in the cell viability （P<0.01）， elevations in the levels of ET-1， TNF-α， IL-1β， and IL-6 （P<0.01）， declines in both mRNA and protein levels of ZO-1 and claudin-5 （P<0.01）， and rises in mRNA and protein levels of RhoA and ROCK2 （P<0.01）. Compared with the model group， the Shaoyaotang-containing serum （10%， 15%， and 20%） groups had enhanced cell viability （P<0.01）， lowered levels of ET-1， TNF-α， IL-1β， and IL-6 （P<0.01）， up-regulated mRNA and protein levels of ZO-1 and claudin-5 （P<0.05， P<0.01）， and down-regulated mRNA and protein levels of RhoA and ROCK2 （P<0.01）. Moreover， the inhibitor group and the 15% and 20% Shaoyaotang-containing serum groups had lower levels of ET-1， TNF-α， IL-1β， and IL-6 （P<0.05， P<0.01）， higher mRNA and protein levels of ZO-1 and claudin-5 （P<0.05， P<0.01）， and lower mRNA and protein levels of RhoA and ROCK2 （P<0.05， P<0.01） than the 10% Shaoyaotang-containing serum group. ConclusionThe Shaoyaotang-containing serum can lower the levels of LPS-induced increases in levels of inflammatory cytokines and endothelin to ameliorate the damage of tight junction proteins of the Caco-2 cell model of inflammation by regulating the expression of proteins in the RhoA/ROCK pathway.

ObjectiveTo investigate the effects of different concentrations of Shaoyaotang-containing serum on lipopolysaccharide （LPS）-induced inflammation of human colorectal adenocarcinoma （Caco-2） cells by inhibiting apoptosis via activating the tumor necrosis factor （TNF） receptor superfamily member 6 （Fas）/Fas ligand （FasL） pathway. MethodsCaco-2 cells were allocated into blank， model （LPS， 10 mg·L^-1）， Shaoyaotang-containing serum （5%， 10%， 15%， 20%）， and Fas inhibitor （KR-33493， 20 mmol·L^-1） groups. Except the blank group， the other groups were stimulated with 10 mg·L^-1 LPS for 24 h for the modeling of inflammation. After successful modeling， the blank， Fas inhibitor， and model groups were treated with blank serum， and the Shaoyaotang-containing serum groups were treated with the serum samples at corresponding concentrations for 24 h. The Fas inhibitor group was subjected to KR-33493 pretreatment for 1 h. Cell proliferation and viability were examined by the cell-counting kit-8 （CCK-8） method. The levels of interleukin （IL）-6， IL-1β， and TNF-α were measured by enzyme-linked immunosorbent assay. Apoptosis was detected by flow cytometry. The protein and mRNA levels of Fas， FasL， cysteinyl aspartate-specific proteinase （Caspase）-3， Caspase-9， B-cell lymphoma 2 （Bcl-2）， and Bcl-2-associated X protein （Bax） were determined by Western blot and Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）， respectively. ResultsCompared with the blank group， the model group presented a decrease in cell survival rate （P<0.01）. Compared with that in the model group， the cell survival rate showed no significant change in the 5% Shaoyaotang-containing serum group but increased in the 10%， 15%， and 20% Shaoyaotang-containing serum groups （P<0.01）. Since there was no statistical difference between the 5% Shaoyaotang-containing serum group and the model group， 10%， 15%， and 20% Shaoyaotang-containing sera were selected for the follow-up study. Compared with the blank group， the model group showed risen levels of IL-6， IL-1β， and TNF-α （P<0.01）， an increased apoptosis rate （P<0.01）， up-regulated protein and mRNA levels of Fas， FasL， Caspase-3， Caspase-9， and Bax （P<0.01）， and down-regulated protein and mRNA levels of Bcl-2 （P<0.01）. Compared with the model group， the Fas inhibitor group and the 10%， 15%， and 20% Shaoyaotang-containing serum groups showed declined levels of IL-6， IL-1β， and TNF-α （P<0.01）， decreased apoptosis rates （P<0.01）， down-regulated protein and mRNA levels of Fas， FasL， Caspase-3， Caspase-9， and Bax （P<0.05， P<0.01）， and up-regulated protein and mRNA levels of Bcl-2 （P<0.05， P<0.01）. In addition， the 15% and 20% Shaoyaotang-containing serum groups had lower levels of IL-6， IL-1β， and TNF-α （P<0.05， P<0.01）， lower apoptosis rates （P<0.05， P<0.01）， lower protein and mRNA levels of Fas， FasL， Caspase-3， Caspase-9， and Bax （P<0.05， P<0.01）， and higher protein and mRNA levels of Bcl-2 （P<0.05， P<0.01） than the 10% Shaoyaotang-containing serum group. ConclusionThe Shaoyaotang-containing serum can reduce the content of inflammatory factors in Caco-2 cells， down-regulate the protein and mRNA levels of Fas， FasL， Caspase-3， Caspase-9， and Bax， and up-regulate the protein and mRNA levels of Bcl-2 under the intervention of LPS by regulating the Fas/FasL pathway and inhibiting the apoptosis of intestinal epithelial cells in ulcerative colitis.

ObjectiveTo investigate the protective effect and mechanism of Shaoyaotang （SYD） on the lipopolysaccharide （LPS）-induced damage of tight junction proteins in the human colorectal adenocarcinoma （Caco-2） cell model of inflammation via the Ras homolog gene family member A （RhoA）/Rho-associated coiled-coil forming protein kinase （ROCK） pathway. MethodsCaco-2 cells were grouped as follows： Blank， model （LPS， 10 mg·L^-1）， SYD-containing serum （10%， 15%， and 20%）， and inhibitor （Fasudil， 25 μmol·L^-1）. After 24 hours of intervention， the cell viability in each group was examined by the cell-counting kit 8 （CCK-8） method. Enzyme-linked immunosorbent assay was employed to determine the levels of endothelin-1 （ET-1）， tumor necrosis factor-α （TNF-α）， interleukin-1β （IL-1β）， and interleukin-6 （IL-6）. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot were employed to determine the mRNA and protein levels， respectively， of RhoA， ROCK2， claudin-5， and zonula occludens-1 （ZO-1） in cells of each group. ResultsCompared with the blank group， the model group showcased a marked reduction in the cell viability （P<0.01）， elevations in the levels of ET-1， TNF-α， IL-1β， and IL-6 （P<0.01）， declines in both mRNA and protein levels of ZO-1 and claudin-5 （P<0.01）， and rises in mRNA and protein levels of RhoA and ROCK2 （P<0.01）. Compared with the model group， the Shaoyaotang-containing serum （10%， 15%， and 20%） groups had enhanced cell viability （P<0.01）， lowered levels of ET-1， TNF-α， IL-1β， and IL-6 （P<0.01）， up-regulated mRNA and protein levels of ZO-1 and claudin-5 （P<0.05， P<0.01）， and down-regulated mRNA and protein levels of RhoA and ROCK2 （P<0.01）. Moreover， the inhibitor group and the 15% and 20% Shaoyaotang-containing serum groups had lower levels of ET-1， TNF-α， IL-1β， and IL-6 （P<0.05， P<0.01）， higher mRNA and protein levels of ZO-1 and claudin-5 （P<0.05， P<0.01）， and lower mRNA and protein levels of RhoA and ROCK2 （P<0.05， P<0.01） than the 10% Shaoyaotang-containing serum group. ConclusionThe Shaoyaotang-containing serum can lower the levels of LPS-induced increases in levels of inflammatory cytokines and endothelin to ameliorate the damage of tight junction proteins of the Caco-2 cell model of inflammation by regulating the expression of proteins in the RhoA/ROCK pathway.

Objective To develop high-immunogenicity antigens targeting the BA.5 variant of SARS-CoV-2 and to screen for monoclonal antibodies with high neutralizing and blocking activity,providing new strategies for the development of vaccines and diagnostic reagents.Methods The ZP protein gene fragment from zebrafish vitel-logenin and the HIS protein tag were inserted into the pCDNA3.4 plasmid to construct the recombinant plasmid pCDNA3.4-RBD(BA.5)-ZP-HIS,which was then transfected into 293F cells to express the RBD-ZP protein.The expression of the protein was verified by SDS-PAGE and its binding capabilities to ACE2 receptor molecules and aluminum adjuvant were detected.The immunogenicity of the fusion protein was evaluated using a BALB/c mouse model,and monoclonal antibodies were prepared through hybridoma technology.Monoclonal antibodies with strong neutralizing and blocking activity were screened and their neutralizing activity was detected by blocking ELISA.Results The ZP gene and HIS protein tag sequence were successfully inserted into the pCDNA3.4 vector and the RBD-ZP protein with a molecular weight of 50 kDa was successfully expressed.The immunogenicity test results showed that the ZP protein effectively enhanced the immunogenicity of the RBD protein and improved its binding capability to the ACE2 receptor.After immunizing mice with the RBD-ZP protein,8 monoclonal antibodies that specifically bind to both the mutant and wild-type strains were cloned and screened through hybridoma technology,among which 3 could effectively block the binding of the SARS-CoV-2 RBD protein to the human ACE2 receptor.Conclusion This study successfully expressed the RBD-ZP fusion protein,which significantly enhanced the immunogenicity and receptor binding capability of the RBD protein.Three monoclonal antibodies with high neutral-izing and blocking activity were screened out,providing strong support for the development of COVID-19 vaccines and diagnostic reagents..

As a physical treatment technique, modified electro-convulsive therapy (MECT) is used to treat mental and certain neurological disorders by causing seizures with short, suitable electrical currents applied to the brain while the patient is under general anesthesia and muscle relaxants. MECT is recognized for its therapeutic efficacy and clinical safety, rendering it one of the most prevalent interventions in psychiatric care. To enhance clinical outcomes and minimize adverse effects, this consensus document delineates the indications, therapeutic parameters, therapeutic procedures, potential adverse effects, and associated management strategies for MECT. These guidelines are informed by the latest clinical research and expert consensus, integrating evidence-based medicine methodologies. The objective is to furnish clinicians with precise operational guidelines and to advance the standardization of MECT practices in clinical settings.