OBJECTIVE To investigate the ameliorative effect and mechanism of Euphorbia hirta L.-derived exosome-like nanovesicles（Eh-ENVs） on acetaminophen （APAP）-induced liver injury based on the nuclear factor erythroid 2 related factor 2 （Nrf2）/heme oxygenase-1 （HO-1）/NAD（P）H：quinone oxidoreductase 1 （NQO1） pathway. METHODS The safety of Eh-ENVs was evaluated by examining their effects on the viability of RAW264.7 and AML12 cells， as well as serum liver and kidney function indicators and histopathology of liver， lung， and other tissues in normal mice. A lipopolysaccharide （1 μg/mL）-induced RAW264.7 cell inflammation model was constructed to investigate the effects of 10 and 20 μg/mL Eh-ENVs on the mRNA expression of inflammatory factors and reactive oxygen species （ROS） level in model cells， and the uptake efficiency of Eh-ENVs by RAW264.7 cells was also examined. An APAP-induced liver injury mouse model was established to investigate the effects of 4 mg/kg Eh-ENVs on serum liver function indicators， liver histopathology， mRNA expression of inflammatory factors， malondialdehyde （MDA） level， superoxide dismutase （SOD） level， and mRNA and protein expressions related to the Nrf2/HO-1/NQO1 pathway in liver tissue of model mice. RESULTS In vitro results showed that Eh-ENVs had no inhibitory effect on the proliferation of RAW264.7 and AML12 cells；Eh-ENVs could be efficiently taken up by RAW264.7 cells and significantly reduced the mRNA expression of interleukin-1β （IL-1β）， tumor necrosis factor-α （TNF-α）， and ROS level in cells （ P ＜0.05）. In vivo results showed that 4 mg/kg Eh-ENVs had no obvious toxic side effects on normal mice，could significantly decrease the serum alanine transaminase （ALT） and aspartate transaminase （AST） levels in model mice （ P ＜0.05），upregulated/increased the mRNA expressions of IL-10， as well as the mRNA and protein expressions of Nrf2， HO-1， and NQO1， and SOD level in liver tissue （ P ＜0.05）， and down-regulated/decreased the mRNA expression of TNF-α， IL-1β and MDA level in liver tissue （ P ＜0.05）. CONCLUSIONS Eh-ENVs may activate the Nrf2/HO-1/NQO1 pathway to inhibit inflammatory response and alleviate oxidative stress， thereby improving APAP-induced liver injury.

ObjectiveTo investigate the mechanism of action of modified Chaihu Shugan Powder in the treatment of abnormal gallbladder relaxation in gallbladder cholesterol stone （CS） with liver depression syndrome， and to provide a basis for clinical medication. MethodsMice were given a high-fat lithogenic diet combined with chronic unpredictable mild stress （CUMS） to establish a model of CS. A total of 45 male C57BL/6 mice were randomly divided into blank group （6 mice fed a normal diet） and CS group （39 mice fed a high-fat lithogenic diet）. After CS modeling， the CS group was further randomly divided into four subgroups of CS group， CS liver depression group， traditional Chinese medicine group （treated with modified Chaihu Shugan Powder）， and Western medicine group （treated with ursodeoxycholic acid）， with 9 mice in each group. All subgroups were fed with the high-fat lithogenic diet， and all mice except those in the CS group were given 21 days of CUMS for modeling. Samples were collected after intervention. The serum levels of cholecystokinin （CCK）， liver function parameters， and blood lipid profiles were measured； HE staining was performed for liver and gallbladder tissue； qPCR and Western blot were used to measure the mRNA and protein expression levels of G protein-coupled bile acid receptor 1 （TGR5） and glucagon-likepeptide-1/2 （GLP-1/2） in the intestine and TGR5 and glucagon-like peptide-2 receptor （GLP-2R） in gallbladder； metabolomics methods were used to determine bile acid composition in intestinal contents. The independent-samples t-test was used for comparison of continuous data between two groups； a one-way analysis of variance was used for comparison between multiple groups， and the least significant difference t-test or the Games-Howell method was used for further comparison between two groups. ResultsCompared with the blank group， the CS group showed significant gallstone formation， bile turbidity， hepatic steatosis， abnormal gallbladder wall structure， and significant increases in anxiety- and depression-like behaviors based on behavioral tests； significant increases in the level of total cholesterol in bile and the serum levels of alanine aminotransferase， aspartate aminotransferase， and low-density lipoprotein and significant reductions in the level of total bile acid （TBA） in bile and the serum levels of CCK and high-density lipoprotein （HDL） （all P<0.05）； significant increases in the mRNA expression levels of GLP-1/2 and TGR5 in the intestine and the protein expression levels of GLP-2R and TGR5 in the gallbladder and significant reductions in the mRNA expression levels of GLP-2R and TGR5 in the gallbladder （all P<0.05）； significant changes in multiple bile acid components in intestinal contents （all P<0.05）. Compared with the CS group， the CS liver depression group had further aggravation of pathological and behavioral manifestations， changes in bile acid composition， significant increases in the protein and mRNA expression levels of TGR5 and GLP-1/2 in the intestine， and significant increases in the protein and mRNA expression levels of TGR5 and GLP-2R in the gallbladder （all P<0.01）. Compared with the CS liver depression group， both treatment groups had an improvement in gallbladder morphology， alleviation of stones and liver injury， and recovery of liver function and blood lipid levels， as well as significant reductions in the protein and mRNA expression levels of TGR5 and GLP-1/2 in the intestine and TGR5 and GLP-2R in the gallbladder （all P<0.05）； the traditional Chinese medicine group showed significant increases in glycodeoxycholic acid （GDCA）， tauro-α-muricholic acid （T-α-MCA）， and taurochenodeoxycholic acid （TCDCA） （all P<0.05）， while the Western medicine group showed significant increases in taurohyodeoxycholic acid， T-α-MCA， TCDCA， GDCA， and glycoursodeoxycholic acid （all P<0.05）. Compared with the Western medicine group， the traditional Chinese medicine group had significantly greater behavioral improvements， significantly higher levels of TBA in bile and serum HDL （both P<0.01）， significant reductions in the protein expression levels of TGR5 and GLP-1/2 in the intestine and TGR5 and GLP-2R in the gallbladder， and a significant reduction in the mRNA expression level of TGR5 in the intestine （all P<0.01）， as well as a significant increase in tauroursodeoxycholic acid and significant reductions in glycoursodeoxycholic acid， taurohyodeoxycholic acid， TCDCA， and taurolithocholic acid （all P<0.05）. ConclusionModified Chaihu Shugan Powder can improve liver function and abnormal gallbladder relaxation in CS with liver depression syndrome by regulating the bile acid-TGR5 axis， thereby exerting the therapeutic effect of soothing the liver， resolving depression， moving Qi， and promoting bile flow.

ObjectiveTo investigate the biological reparative effects of platelet-rich fibrin （PRF） extract on injured endometrial stromal cells （ESCs） induced by mifepristone and the underlying molecular mechanisms. MethodsAn ESCs injury model was established using mifepristone treatment. Cells were divided into four groups： untreated control （Con）， mifepristone-injured group （M）， mifepristone treatment+PRF extract group （M+P）， and mifepristone treatment+PRF extract+growth factor inhibitors treatment group， including M+P+T， M+P+Me， and M+P+CJ. Cell viability was detected using the CCK-8 assay， and migration ability was evaluated via the scratch wound healing assay. The release kinetics of growth factors， including vascular endothelial growth factor （VEGF）， platelet-derived growth factor-BB （PDGF-BB）， and transforming growth factor-beta 1 （TGF-β1） from PRF extract were measured by ELISA. The mRNA and protein expression levels of apoptosis-related factors （Bcl-2， Bax， and Caspase-3） were analyzed by RT-qPCR and Western blot， respectively. ResultsVEGF， PDGF-BB， and TGF-β1 were continuously released from the PRF extract over two weeks， with peak release observed on days 3-7. The extract significantly restored the proliferation and migration of mifepristone-injured ESCs （all P<0.000 1）. PRF extract up-regulated the mRNA and protein expression of the anti-apoptotic factor Bcl-2 （all P<0.000 1） and down-regulated that of the pro-apoptotic factors Bax and Caspase-3 （all P<0.000 1） in mifepristone-injured ESCs. Inhibitors of VEGF， PDGF-BB， and TGF-β1 （Tanshinone IIA， Methylnissolin， and CJJ300） significantly reversed the protective effects of PRF extract on mifepristone-injured ESCs （all P<0.05）. ConclusionPRF extract promotes the repair of injured ESCs by enhancing cell proliferation， attenuating apoptosis， and increasing migration capacity， which involve multiple growth factors， including VEGF， PDGF-BB， and TGF-β1.

Objective To investigate the exposure-response relationships and lag effects between air pollutants (PM_2.5, PM₁₀, O₃, and NO₂) and mortality in Jiading District, Shanghai, and to provide a scientific basis for the formulation of environmental health policies. Methods Using an individual-level time-stratified case-crossover design, conditional logistic regression models in conjunction with a distributed lag nonlinear model (DLNM) were employed to analyze the exposure-response relationship and temporal lag patterns of ambient air pollution on resident mortality in Jiading District (1994–2024). Results A total of 59 048 death cases were collected, including 18,701 deaths from cardiovascular diseases and 11 731 deaths from respiratory diseases. PM_2.5 and NO₂ had a significant impact on all-cause mortality, cardiovascular disease mortality, and respiratory disease mortality, with the most significant effects observed within a lag of 0–3 days. PM₁₀ also had some impact on these three types of mortality, but its effect was generally weaker than that of PM_2.5 and NO₂. The exposure-response curves showed that the risk of death increased rapidly with increasing concentrations of PM_2.5 and PM₁₀, while the effect of NO₂ plateaued at higher levels. No significant differences were found across age or gender subgroups. Conclusion Short-term exposure to PM_2.5, PM₁₀, and NO₂ significantly increases all-cause mortality risk in Jiading District, with effects persisting up to 7 days, highlighting the need for enhanced air pollution control measures, particularly targeting fine particulate matter.

ObjectiveTo investigate the effects of 7-dehydrocholesterol reductase （DHCR7） in promoting proliferation， migration， and invasion of lung adenocarcinoma cells and the underlying signaling mechanisms. MethodsThe expression level of DHCR7 in lung adenocarcinoma and its prognosis were analyzed by using public databases. DHCR7 protein expression levels in the lung adenocarcinoma cell lines H1299 and A549 were assessed by Western blot， using the normal lung epithelial cell line BEAS-2B as a control. Small interfering RNA （siRNA） was used to knock down DHCR7 expression in H1299 and A549 cells. Assays including colony formation， CCK-8， wound healing， and Transwell experiments were conducted to assess the proliferation， migration， and invasion of the knockdown cells. Next， Western blot was employed to assess the phosphorylation levels of key signaling molecules in the phosphoinositide 3-kinase/protein kinase B （PI3K/AKT）， janus kinase/signal transducer and activator of transcription 3 （JAK/STAT3），mitogen-activated protein kinase/extracellular signal-regulated kinase pathway （MAPK/ERK） （p-AKT， p-STAT3， p-ERK）. ResultsBioinformatics results indicated that both DHCR7 mRNA and protein were highly expressed in lung adenocarcinoma compared with normal tissues adjacent to cancer. Furthermore， higher DHCR7 mRNA level was associated with poor prognosis （P<0.001）. The expression levels of DHCR7 were significantly higher in both H1299 and A549 cells than that in BEAS-2B cells （P<0.05， P<0.01）. Compared with si-control， the proliferation， migration， and invasion abilities of si-DHCR7 H1299 and A549 cells significantly decreased. Among the phosphorylated signaling molecules detected， p-ERK was significantly downregulated （P<0.000 1， P<0.01） whereas p-AKT and p-STAT3 levels were not significantly changed. ConclusionDHCR7 has prominent effects in promoting the proliferation， migration， and invasion of lung adenocarcinoma cells， and the underlying mechanisms are related to the MAPK/ERK signaling pathway.

Objective: To investigate the infectious disease investigation and response capabilities of grassroots Centers for Disease Control and Prevention (CDC) in Zhejiang Province, so as to provide a reference for strengthening the development of CDC workforce. Methods: From May to June 2024, professional technical personnel engaged in or participating in on-site epidemiological investigation and response from 11 municipal-level CDC and 91 county-level CDC in Zhejiang Province were selected as survey participants. Data on sociodemographic information, outbreak response experience, and infectious disease investigation capabilities were collected using the National Questionnaire on the Current Status of Field Epidemiology Investigation Teams. Results: A total of 2 316 individuals were surveyed, including 459 from municipal-leve CDC (19.82%) and 1 857 from county-level CDC (80.18%). The cohort was predominantly female, aged 30-<40 years, with a bachelor's degree, specialized in public health and preventive medicine, holding intermediate professional titles, and having <10 years of work experience, accounting for 55.79%, 36.44%, 72.88%, 74.18%, 34.72%, and 53.76%, respectively. A relatively high proportion of participants had experience in on-site infectious disease response (81.39%), been members of field epidemiology investigation teams or emergency response teams responsible for field epidemiology investigation (81.17%), and had prior work experience in infectious disease-related departments (66.80%). In contrast, the proportion of those who had delivered academic presentations on field epidemiology investigations (4.15%) or contributed to the development of expert consensus documents, guidelines, or textbooks (8.51%). The total score for infectious disease investigation and response capabilities among field epidemiologists investigators in grassroots CDC in Zhejiang Province was 53.19±16.19, with dimension scores of 65.15±13.90 for personal comprehensive competence, 59.75±20.77 for outbreak investigation procedures, and 28.65±19.32 for essential epidemiological investigation skills. Conclusions The grassroots field epidemiology investigation teams in Zhejiang Province have developed a certain level of workforce capacity and outbreak response experience. However, they generally exhibit relatively short work tenures, limited academic outputs, and moderate-to-low proficiency in infectious disease investigation and management, particularly with notable gaps in critical field investigation skills. It is recommended to strengthen systematic training and practical exercises to enhance grassroots emergency response capabilities.

Obstructive sleep apnea-hypopnea syndrome(OSAHS)is a chronic disease that characteristics is multi system damage,which core pathological mechanism involves a complex interaction between structural abnormalities of the upper airway and imbalances in neuromuscular regulation.With continuous advancements in imaging technique,upper airway imaging technique has been a key tool that reveals pathological characteristics of OSAHS heterogeneity,and guides individual diagnosis and treatment.This article explored OSAHS's pathogenesis,and systematically sorted out the application of various imaging techniques,including X-ray cephalometry,computed tomography(CT),magnetic resonance imaging(MRI),ultrasound,and drug-induced sleep endoscopy(DISE)in the diagnosis and treatment for OSAHS,which focuses on an analysis for their values in the location of them at obstructive plane,treatment decision and the prediction for curative effect.In addition,this article conducted a prospect for multimodal imaging fusion,artificial intelligence-assisted analysis and other development in these imaging techniques in future.By comparing the clinical applicability and limitations of each kind of technique,this review provided imaging basis for optimizing the pathway of OSAHS's clinical diagnosis and treatment,promoting the shift from conventional classification to precision intervention,so as to ultimately improve the quality of managing disease.

Hearing loss is the most prevalent disabling disease. Cochlear implantation（CI） serves as the primary intervention for severe to profound hearing loss. This consensus systematically explores the value of genetic diagnosis in the pre-operative assessment and efficacy prognosis for CI. Drawing upon domestic and international research and clinical experience, it proposes an evidence-based medicine three-tiered prognostic classification system（Favorable, Marginal, Poor）. The consensus focuses on common hereditary non-syndromic hearing loss（such as that caused by mutations in genes like GJB2, SLC26A4, OTOF, LOXHD1） and syndromic hereditary hearing loss（such as Jervell & Lange-Nielsen syndrome and Waardenburg syndrome）, which are closely associated with congenital hearing loss, analyzing the impact of their pathological mechanisms on CI outcomes. The consensus provides recommendations based on multiple round of expert discussion and voting. It emphasizes that genetic diagnosis can optimize patient selection, predict prognosis, guide post-operative rehabilitation, offer stratified management strategies for patients with different genotypes, and advance the application of precision medicine in the field of CI.
Humans ; Cochlear Implantation ; Prognosis ; Hearing Loss/surgery* ; Consensus ; Connexin 26 ; Mutation ; Sulfate Transporters ; Connexins/genetics*

Objective To investigate the changes in eosinophil counts and the activation of microglial cells in the brain tissues of mice at different stages of Angiostrongylus cantonensis infection, and to examine the role of microglia in regulating the progression of angiostrongyliasis and unravel the possible molecular mechanisms. Methods Fifty BALB/c mice were randomly divided into the control group and the 7-d, 14-d, 21-day and 25-d infection groups, of 10 mice in each group. All mice in infection groups were infected with 30 stage III A. cantonensis larvae by gavage, and animals in the control group was given an equal amount of physiological saline. Five mice were collected from each of infection groups on days 7, 14, 21 d and 25 d post-infection, and 5 mice were collected from the control group on the day of oral gavage. The general and focal functional impairment was scored using the Clark scoring method to assess the degree of mouse neurological impairment. Five mice from each of infection groups were sacrificed on days 7, 14, 21 d and 25 d post-infection, and 5 mice from the control group were sacrificed on the day of oral gavage. Mouse brain tissues were sampled, and the pathological changes of brain tissues were dynamically observed using hematoxylin and eosin (HE) staining. Immunofluorescence staining with eosinophilic cationic protein (ECP) and ionized calcium binding adaptor molecule 1 (Iba1) was used to assess the degree of eosinophil infiltration and the counts of microglial cells in mouse brain tissues in each group, and the morphological parameters of microglial cells (skeleton analysis and fractal analysis) were quantified by using Image J software to determine the morphological changes of microglial cells. In addition, the expression of M1 microglia markers Fcγ receptor III (Fcgr3), Fcγ receptor IIb (Fcgr2b) and CD86 antigen (Cd86), M2 microglia markers Arginase 1 (Arg1), macrophage mannose receptor C-type 1 (Mrc1), chitinase-like 3 (Chil3), and phagocytosis genes myeloid cell triggering receptor expressed on myeloid cells 2 (Trem2), CD68 antigen (Cd68), and apolipoprotein E (Apoe) was quantified using real-time quantitative reverse transcription PCR (RT-qPCR) assay in the mouse cerebral cortex of mice post-infection. Results A large number of A. cantonensis larvae were seen on the mouse meninges surface post-infection, and many neuronal nuclei were crumpled and deeply stained, with a large number of bleeding points in the meninges. The median Clark scores of mouse general functional impairment were 0 (interquartile range, 0), 0 (interquartile range, 0.5), 6 (interquartile range, 1.0), 14 (interquartile range, 8.5) points and 20 (interquartile range, 9.0) points in the control group and the 7-d, 14-d, 21-d and 25-d groups, respectively (H = 22.45, P < 0.01), and the median Clark scores of mouse focal functional impairment were 0 (interquartile range, 0), 2 (interquartile range, 2.5), 7 (interquartile range, 3.0), 18 (interquartile range, 5.0) points and 25 (interquartile range, 6.5) points in the control group and the 7-d, 14-d, 21-d and 25-d groups, respectively (H = 22.72, P < 0.01). The mean scores of mice general and focal functional impairment were all higher in the infection groups than in the control group (all P values < 0.05). Immunofluorescence staining showed a significant difference in the eosinophil counts in mouse brain tissues among the five groups (F = 40.05, P < 0.000 1), and the eosinophil counts were significantly higher in mouse brain tissues in the 14-d (3.08 ± 0.78) and 21-d infection groups (5.97 ± 1.37) than in the control group (1.00 ± 0.28) (both P values < 0.05). Semi-quantitative analysis of microglia immunofluorescence showed a significant difference in the counts of microglial cells among the five groups (F = 17.66, P < 0.000 1), and higher Iba1 levels were detected in mouse brain tissues in 14-d (5.75 ± 1.28), 21-d (6.23 ± 1.89) and 25-d infection groups (3.70 ± 1.30) than in the control group (1.00 ± 0.30) (all P values < 0.05). Skeleton and fractal analyses showed that the branch length [(162.04 ± 34.10) μm vs. (395.37 ± 64.11) μm; t = 5.566, P < 0.05] and fractal dimension of microglial cells (1.30 ± 0.01 vs. 1.41 ± 0.03; t = 5.266, P < 0.05) were reduced in mouse brain tissues in the 21-d infection group relative to the control group. In addition, there were significant differences among the 5 groups in terms of M1 and M2 microglia markers Fcgr3 (F = 48.34, P < 0.05), Fcgr2b (F = 55.46, P < 0.05), Cd86 (F = 24.44, P < 0.05), Arg1 (F = 31.18, P < 0.05), Mrc1 (F = 15.42, P < 0.05) and Chil3 (F = 24.41, P < 0.05), as well as phagocytosis markers Trem2 (F = 21.19, P < 0.05), Cd68 (F = 43.95, P < 0.05) and Apoe (F = 7.12, P < 0.05) in mice brain tissues. Conclusions A. cantonensis infections may induce severe pathological injuries in mouse brain tissues that are characterized by massive eosinophil infiltration and persistent activation of microglia cells, thereby resulting in progressive deterioration of neurological functions.

Objective To analyze the underestimated influencing factors of ductal carcinoma in situ(DCIS)detected by ultrasound-guided fine needle aspiration cytology(US-FNAC),and to construct and validate the nomogram prediction model.Methods A total of 105 patients with DCIS diagnosed by US-FNAC in Yixing Hospital of Traditional Chinese Medicine from January 2020 to June 2023 were retrospectively selected.All patients were female and aged 40-76 years.According to postoperative pathological results,they were assigned to underestimated group(n=27)and non-underestimated group(n=78).The clinical data of the patients were collected to analyze the influencing factors of the underestimation of DCIS,and a nomogram prediction model was constructed.The receiver operating characteristic curve(ROC)was drawn,and the area under the curve(AUC)was used to analyze the predictive efficacy of the prediction model for the underestimated risk of DCIS before operation.Results Univariate analysis showed that the proportions of palpable masses,tumor size>2 cm,microcalcification,and breast imaging reporting and data system(BI-RADS)4B-5 in the underestimated group were higher than those in the non-underestimated group(P<0.05).Binary logistic regression analysis showed that tumor size[(odds ratio,OR)=4.453,95%(confidence interval,C I):1.890-10.486],microcalcification(OR=3.079,95%CI:1.650-5.742),BI-RADS classification(OR=5.211,95%CI:2.528-10.740)were the independent risk factors for preoperative underestimation of DCIS(P<0.05).Internal validation of the nomogram prediction model based on the above factors showed that the C-index was 0.865(95%CI:0.785-0.952),and the correction curve for predicting the underestimation of DCIS was close to the ideal curve(P>0.05).ROC curve showed that the nomogram model had a sensitivity of 87.90%,a specificity of 88.60%,and an AUC of 0.895(95%CI:0.823-0.966).Conclusion Tumor size,microcalcification and BI-RADS classification are independent risk factors for preoperative underestimation of DCIS.The nomogram prediction model based on these factors can better evaluate the risk of preoperative underestimation of DCIS.