Objective:To investigate the role of hypoxia-induced angiopoietin-like protein 4 (ANGPTL4) expression in experimental choroidal neovascularization (CNV).Methods:Twenty-seven SPF male C57BL/6J mice aged 6-8 weeks were selected.Eighteen of the mice were used to establish a laser-induced CNV model.On the 7th day after laser photocoagulation, success of the modeling was verified using optical coherence tomography angiography (OCTA) and fundus fluorescein angiography (FFA). The retinal pigment epithelium (RPE)-choroid-sclera complex was extracted for protein analysis before modeling and on the 3rd and 7th days after modeling.The relative expression levels of ANGPTL4 and vascular endothelial growth factor (VEGF) at different time points were detected by Western blot.Additionally, frozen sections of mouse eyeballs on day 7 after modeling were prepared and the expression and cellular localization of ANGPTL4 were observed by immunofluorescence.RF/6A cells, derived from monkey choroidal retinal endothelial cells, were treated with 200 μmol/L cobalt chloride (CoCl 2) in the culture medium for 0, 3, 6, and 12 hours.RF/6A cells were also divided into a normal control group, a hypoxia group, and a hypoxia+ si-ANGPTL4 group, and cells were transfected with a plasmid containing si-ANGPTL4 sequence.The relative expression levels of ANGPTL4 and VEGF proteins in each group were detected by Western blot, and the differences in tube formation among the groups were observed by tube formation assay.A total of 27 male C57BL/6J mice were randomly divided into CNV group, CNV+ si-NC group, and CNV+ si-ANGPTL4 group, with 9 mice in each group.In the CNV+ si-NC and CNV+ si-ANGPTL4 groups, si-NC and si-ANGPTL4 were respectively injected into the vitreous cavity after the CNV model was established.Fluorescence leakage in mice was observed by FFA, and the length, thickness and area of CNV was observed using OCTA and immunofluorescence staining of choroidal flat mounts.The relative expression levels of ANGPTL4 and VEGF proteins in each group were detected by Western blot.All animal experiments were conducted in accordance with ARVO Statement on the Use of Animals in Ophthalmic and Vision Research.The experimental protocol was approved by the Affiliated Hospital of Nantong University (No.S20220822-902). Results:Before modeling and on the 3rd and 7th days after modeling, the relative expression levels of ANGPTL4 protein were 1.00±0.00, 1.58±0.05, and 1.90±0.04, respectively, and the relative expression levels of VEGF protein were 1.00±0.00, 1.31±0.05, and 1.84±0.04, respectively, with statistically significant overall differences ( F=528.934, 390.424, both P<0.05). Among them, on the 3rd and 7th days after modeling, the relative expression levels of ANGPTL4 and VEGF proteins were significantly higher in CNV group than in the control group (all P<0.05). The tissues of each layer of the retina were clear in the control group, while neovascularization could be seen growing under the retinal neuroepithelial layer in the CNV group.Compared with the control group, ANGPTL4 expression was significantly increased and colocalized with vascular endothelial cells in the CNV group.After CoCl 2 treatment of RF/6A cells for 3, 6, and 12 hours, the relative expression levels of ANGPTL4 and VEGF proteins were higher than at 0 hour, with statistically significant differences (all P<0.05). Compared with the control group, the relative ANGPTL4 protein expression was increased in the hypoxia group and significantly decreased in the hypoxia+ si-ANGPTL4 group, showing statistically significant differences (both P<0.05). The number of tube formations in the control group, hypoxia group, and hypoxia+ si-ANGPTL4 group were 12.67±1.53, 19.64±1.56, and 17.01±1.04, respectively, with a statistically significant overall difference ( F=33.091, P<0.01). The number of tube formations increased in the hypoxia group and hypoxia+ si-ANGPTL4 group compared with the control group, and the number of tube formations decreased in the hypoxia+ si-ANGPTL4 group compared with the hypoxia group, with statistically significant differences (all P<0.05). Relative expression levels of ANGPTL4 and VEGF proteins were significantly lower in the CNV+ si-ANGPTL4 group than those in the CNV group (both P<0.05). The CNV area was significantly lower in the CNV+ si-ANGPTL4 group than in the CNV group and CNV+ si-NC group (both P<0.05). Conclusions:Hypoxia-induced ANGPTL4 promotes experimental CNV formation in vivo and in vitro.Inhibiting ANGPTL4 can reduce CNV formation and leakage.

OBJECTIVE To investigate the role of δ-subunit-containing extrasynaptic γ-aminobutyric acid type A receptor(GABAAR)in sleep-promoting effects of zolpidem(ZPD).METHODS ①C57BL/6J mice were implanted with skull electrodes and allowed postoperative recovery of seven days.Groups of mice were intraperitoneally(ip)administered with ZPD at 0(vehicle control),2.5,5 and 10 mg·kg-1.Cortical electroencephalography(EEG)was recorded to analyze latencies of non-rapid eye movement(NREM)and rapid eye movement(REM)sleep,the percentage of wakefulness,NREM and REM sleep,sleep architecture(the proportion of NREM and REM sleep in sleep,number of times and mean duration of NREM and REM sleep,microarousals and short awakenings),EEG power density and slow-wave activity(SWA,0.5-4 Hz)during NREM sleep.② Wild-type(WT)and δ-subunit knockout(δ-KO)mice were ip administered with vehicle or ZPD 10 mg·kg-1 before cortical EEG parameters were ana-lyzed to compare ZPD effects between genotypes.RESULTS ①Compared with the vehicle,ZPD 2.5,5 and 10 mg·kg-1 significantly shortened NREM sleep latency.ZPD 5 and 10 mg·kg-1 markedly reduced wakefulness and increased NREM sleep time.ZPD 2.5 and 10 mg·kg-1 increased NREM sleep episode frequency while ZPD 10 mg·kg-1 elevated brief awakening frequency.REM sleep remained unchanged.②In δ-KO mice,ZPD 10 mg·kg-1 significantly shortened NREM sleep latency compared with WT mice,but its effects on increasing short awakenings and suppressing SWA were abolished.Zolpidem showed no significant differences in the proportion of each sleep phase,the average duration of NREM sleep,and the frequency of REM sleep in KO mice compared to its effects on WT mice.CONCLUSION ZPD-induced sleep fragmentation and reduced sleep depth are mediated by δ-subunit-containing extra-synaptic GABAAR,whereas its shortening of NREM sleep latency is independent of this receptor subtype.

Objective:To evaluate the feasibility of training physicians from secondary comprehensive hospitals in the clinical assessment of brain death and to provide recommendations for nationwide implementation.Methods:This prospective cohort study enrolled physicians who completed standardized training in clinical brain death determination at five pilot hospitals between June and December 2023. Participants were from internal medicine, neurology, critical care, emergency, or anesthesiology departments of secondary comprehensive hospitals and had ≥5 years of clinical experience. Organ donation coordinators and surgeons involved in organ donation or transplantation were excluded. The training program comprised four modules: didactic lectures, bedside demonstrations, simulation-based practice, and written theoretical assessment with review. The theoretical assessment was considered qualified if the score was 60 or above. Participants were categorized into ≥80 and <80 groups based on assessment scores. Between-group comparisons were conducted using rank-sum or chi-square tests.Results:A total of 191 physicians from 74 secondary comprehensive hospitals were enrolled. Most held a bachelor's degree [89.5%(171/191)] and had intermediate [47.1%(90/191)] or associate senior [36.1%(69/191)] professional titles; [59.7%(114/191)] were from non-neurology specialties. The overall pass rate was 99.5% (190/191), with a mean score of 82.4±7.1. Compared with those scoring<80 (56 participants), physicians scoring ≥80 (135 participants) differed significantly by professional title, province, and department ( P=0.014, 0.019 and 0.039). The proportion scoring<80 was higher among junior/intermediate versus senior titles [38.0%(41/108) vs 18.1%(15/83), P=0.003), and among non-neurology/critical care departments (emergency, internal medicine, anesthesiology) versus neurology/critical care [39.7%(31/78) vs 22.1%(25/113), P=0.009]. Only 2.09%(4/191) achieved a perfect score. Across all test items, the overall error rate was 14.99%(700/4 670). The five knowledge points with the highest error rates were mistriggering of mechanical ventilation [96.97%(32/33)], corneal reflex [42.25%(30/71)], spinal reflexes [24.25%(65/268)], documentation of the determination [21.21%(7/33)], and the apnea test procedure [20.73%(57/275)]. Conclusions:The pilot hospitals can effectively deliver clinical training for brain death determination, supporting nationwide promotion. However, physicians' theoretical grounding in neurology at secondary comprehensive hospitals appears relatively weak. Training curricula should be optimized to further improve training quality.

Objective:To investigate the role of hypoxia-induced angiopoietin-like protein 4 (ANGPTL4) expression in experimental choroidal neovascularization (CNV).Methods:Twenty-seven SPF male C57BL/6J mice aged 6-8 weeks were selected.Eighteen of the mice were used to establish a laser-induced CNV model.On the 7th day after laser photocoagulation, success of the modeling was verified using optical coherence tomography angiography (OCTA) and fundus fluorescein angiography (FFA). The retinal pigment epithelium (RPE)-choroid-sclera complex was extracted for protein analysis before modeling and on the 3rd and 7th days after modeling.The relative expression levels of ANGPTL4 and vascular endothelial growth factor (VEGF) at different time points were detected by Western blot.Additionally, frozen sections of mouse eyeballs on day 7 after modeling were prepared and the expression and cellular localization of ANGPTL4 were observed by immunofluorescence.RF/6A cells, derived from monkey choroidal retinal endothelial cells, were treated with 200 μmol/L cobalt chloride (CoCl 2) in the culture medium for 0, 3, 6, and 12 hours.RF/6A cells were also divided into a normal control group, a hypoxia group, and a hypoxia+ si-ANGPTL4 group, and cells were transfected with a plasmid containing si-ANGPTL4 sequence.The relative expression levels of ANGPTL4 and VEGF proteins in each group were detected by Western blot, and the differences in tube formation among the groups were observed by tube formation assay.A total of 27 male C57BL/6J mice were randomly divided into CNV group, CNV+ si-NC group, and CNV+ si-ANGPTL4 group, with 9 mice in each group.In the CNV+ si-NC and CNV+ si-ANGPTL4 groups, si-NC and si-ANGPTL4 were respectively injected into the vitreous cavity after the CNV model was established.Fluorescence leakage in mice was observed by FFA, and the length, thickness and area of CNV was observed using OCTA and immunofluorescence staining of choroidal flat mounts.The relative expression levels of ANGPTL4 and VEGF proteins in each group were detected by Western blot.All animal experiments were conducted in accordance with ARVO Statement on the Use of Animals in Ophthalmic and Vision Research.The experimental protocol was approved by the Affiliated Hospital of Nantong University (No.S20220822-902). Results:Before modeling and on the 3rd and 7th days after modeling, the relative expression levels of ANGPTL4 protein were 1.00±0.00, 1.58±0.05, and 1.90±0.04, respectively, and the relative expression levels of VEGF protein were 1.00±0.00, 1.31±0.05, and 1.84±0.04, respectively, with statistically significant overall differences ( F=528.934, 390.424, both P<0.05). Among them, on the 3rd and 7th days after modeling, the relative expression levels of ANGPTL4 and VEGF proteins were significantly higher in CNV group than in the control group (all P<0.05). The tissues of each layer of the retina were clear in the control group, while neovascularization could be seen growing under the retinal neuroepithelial layer in the CNV group.Compared with the control group, ANGPTL4 expression was significantly increased and colocalized with vascular endothelial cells in the CNV group.After CoCl 2 treatment of RF/6A cells for 3, 6, and 12 hours, the relative expression levels of ANGPTL4 and VEGF proteins were higher than at 0 hour, with statistically significant differences (all P<0.05). Compared with the control group, the relative ANGPTL4 protein expression was increased in the hypoxia group and significantly decreased in the hypoxia+ si-ANGPTL4 group, showing statistically significant differences (both P<0.05). The number of tube formations in the control group, hypoxia group, and hypoxia+ si-ANGPTL4 group were 12.67±1.53, 19.64±1.56, and 17.01±1.04, respectively, with a statistically significant overall difference ( F=33.091, P<0.01). The number of tube formations increased in the hypoxia group and hypoxia+ si-ANGPTL4 group compared with the control group, and the number of tube formations decreased in the hypoxia+ si-ANGPTL4 group compared with the hypoxia group, with statistically significant differences (all P<0.05). Relative expression levels of ANGPTL4 and VEGF proteins were significantly lower in the CNV+ si-ANGPTL4 group than those in the CNV group (both P<0.05). The CNV area was significantly lower in the CNV+ si-ANGPTL4 group than in the CNV group and CNV+ si-NC group (both P<0.05). Conclusions:Hypoxia-induced ANGPTL4 promotes experimental CNV formation in vivo and in vitro.Inhibiting ANGPTL4 can reduce CNV formation and leakage.

OBJECTIVE To investigate the role of δ-subunit-containing extrasynaptic γ-aminobutyric acid type A receptor(GABAAR)in sleep-promoting effects of zolpidem(ZPD).METHODS ①C57BL/6J mice were implanted with skull electrodes and allowed postoperative recovery of seven days.Groups of mice were intraperitoneally(ip)administered with ZPD at 0(vehicle control),2.5,5 and 10 mg·kg-1.Cortical electroencephalography(EEG)was recorded to analyze latencies of non-rapid eye movement(NREM)and rapid eye movement(REM)sleep,the percentage of wakefulness,NREM and REM sleep,sleep architecture(the proportion of NREM and REM sleep in sleep,number of times and mean duration of NREM and REM sleep,microarousals and short awakenings),EEG power density and slow-wave activity(SWA,0.5-4 Hz)during NREM sleep.② Wild-type(WT)and δ-subunit knockout(δ-KO)mice were ip administered with vehicle or ZPD 10 mg·kg-1 before cortical EEG parameters were ana-lyzed to compare ZPD effects between genotypes.RESULTS ①Compared with the vehicle,ZPD 2.5,5 and 10 mg·kg-1 significantly shortened NREM sleep latency.ZPD 5 and 10 mg·kg-1 markedly reduced wakefulness and increased NREM sleep time.ZPD 2.5 and 10 mg·kg-1 increased NREM sleep episode frequency while ZPD 10 mg·kg-1 elevated brief awakening frequency.REM sleep remained unchanged.②In δ-KO mice,ZPD 10 mg·kg-1 significantly shortened NREM sleep latency compared with WT mice,but its effects on increasing short awakenings and suppressing SWA were abolished.Zolpidem showed no significant differences in the proportion of each sleep phase,the average duration of NREM sleep,and the frequency of REM sleep in KO mice compared to its effects on WT mice.CONCLUSION ZPD-induced sleep fragmentation and reduced sleep depth are mediated by δ-subunit-containing extra-synaptic GABAAR,whereas its shortening of NREM sleep latency is independent of this receptor subtype.

Objective:To evaluate the feasibility of training physicians from secondary comprehensive hospitals in the clinical assessment of brain death and to provide recommendations for nationwide implementation.Methods:This prospective cohort study enrolled physicians who completed standardized training in clinical brain death determination at five pilot hospitals between June and December 2023. Participants were from internal medicine, neurology, critical care, emergency, or anesthesiology departments of secondary comprehensive hospitals and had ≥5 years of clinical experience. Organ donation coordinators and surgeons involved in organ donation or transplantation were excluded. The training program comprised four modules: didactic lectures, bedside demonstrations, simulation-based practice, and written theoretical assessment with review. The theoretical assessment was considered qualified if the score was 60 or above. Participants were categorized into ≥80 and <80 groups based on assessment scores. Between-group comparisons were conducted using rank-sum or chi-square tests.Results:A total of 191 physicians from 74 secondary comprehensive hospitals were enrolled. Most held a bachelor's degree [89.5%(171/191)] and had intermediate [47.1%(90/191)] or associate senior [36.1%(69/191)] professional titles; [59.7%(114/191)] were from non-neurology specialties. The overall pass rate was 99.5% (190/191), with a mean score of 82.4±7.1. Compared with those scoring<80 (56 participants), physicians scoring ≥80 (135 participants) differed significantly by professional title, province, and department ( P=0.014, 0.019 and 0.039). The proportion scoring<80 was higher among junior/intermediate versus senior titles [38.0%(41/108) vs 18.1%(15/83), P=0.003), and among non-neurology/critical care departments (emergency, internal medicine, anesthesiology) versus neurology/critical care [39.7%(31/78) vs 22.1%(25/113), P=0.009]. Only 2.09%(4/191) achieved a perfect score. Across all test items, the overall error rate was 14.99%(700/4 670). The five knowledge points with the highest error rates were mistriggering of mechanical ventilation [96.97%(32/33)], corneal reflex [42.25%(30/71)], spinal reflexes [24.25%(65/268)], documentation of the determination [21.21%(7/33)], and the apnea test procedure [20.73%(57/275)]. Conclusions:The pilot hospitals can effectively deliver clinical training for brain death determination, supporting nationwide promotion. However, physicians' theoretical grounding in neurology at secondary comprehensive hospitals appears relatively weak. Training curricula should be optimized to further improve training quality.

ObjectiveTo investigate the effect of Yishen Tongluo prescription （YSTLP） on apoptosis of renal tubular epithelial cells and explore the mechanism based on endoplasmic reticulum stress pathway of protein kinase R-like endoplasmic reticulum kinase （PERK）/activating transcription factor 4 （ATF4）/transcription factor C/EBP homologous protein （CHOP）. MethodThe db/db mice were randomly divided into model group， valsartan group （10 mg·kg^-1）， and low， middle， high-dose YSTLP groups （1， 2.5， 5 g·kg^-1）. Samples were collected after eight weeks of drug intervention. In addition， db/m mice in the same litter served as the control group. Human renal tubular epithelial cells （HK-2） were cultured in vitro and divided into the control group， advanced glycated end-product （AGE） group， and AGE + low， middle， and high-dose YSTLP groups （100， 200， 400 mg·L^-1）. TdT-mediated dUTP nick end labeling （TUNEL） staining was used to detect the apoptosis rate of HK-2 cells. Methylthiazolyldiphenyl-tetrazolium bromide （MTT） assay was conducted to detect the viability of HK-2 cells. Calcium fluorescence probe staining and luciferase reporter gene method were adopted to detect the luciferase activity of folded protein response element （UPRE） and endoplasmic reticulum stress. Immunohistochemical （IHC） analysis was carried out to measure the protein expressions of phosphorylated PKR （p-PERK）， CHOP， and ATF4. Real-time polymerase chain reaction （Real-time PCR） was used to measure the mRNA expression levels of CHOP and X-box binding protein 1 （XBP1） in mouse kidney and HK-2 cells. Western blot was used to detect the protein expression level of p-PERK， PERK， CHOP， ATF4， B-cell lymphoma-2 （Bcl-2）， Bcl-2 associated X protein （Bax）， and cleaved Caspase-3 in mouse kidney and HK-2 cells. ResultIn the cellular assay， HK-2 cell viability was significantly reduced， and the apoptosis rate was elevated in the AGE group compared with the control group （P<0.01）. The mRNA and protein expression levels of apoptosis-related factor Bcl-2 were significantly reduced （P<0.01）， and those of Bax were significantly increased （P<0.01）. The protein expression level of cleaved Caspase-3 was significantly increased （P<0.01）. Compared with the AGE group， YSTLP administration treatment resulted in elevated cell viability and reduced apoptosis rate （P<0.01）. The mRNA and protein expression levels of Bcl-2 were significantly elevated in a time- and dose-dependent manner （P<0.01）， and those of Bax were significantly reduced in a time- and dose-dependent manner. The protein expression level of cleaved Caspase-3 was significantly reduced in a time- and dose-dependent manner （P<0.01）. The intracellular Ca²⁺ imbalance and UPRE luciferase fluorescence intensity were increased in the AGE group compared with the control group （P<0.01）. The mRNA levels of endoplasmic reticulum stress-related factors CHOP and XBP1 were significantly increased （P<0.01）， and the protein expression levels of p-PERK， CHOP， and ATF4 were significantly increased （P<0.05）. Compared with the AGE group， YSTLP effectively improved intracellular Ca²⁺ imbalance in HK-2 cells and decreased UPRE luciferase fluorescence intensity in a dose-dependent manner （P<0.01）. It reduced the mRNA levels of endoplasmic reticulum stress-related factors CHOP and XBP1 （P<0.01） and the protein expression levels of intracellular p-PERK， CHOP， and ATF4 in a dose- and time-dependent manner （P<0.01）. In animal experiments， the protein expression level of Bcl-2 was significantly reduced（P<0.01）， and that of cleaved Caspase-3 and Bax was significantly increased in the model group compared with the control group （P<0.05）. The protein expression level of Bcl-2 was dose-dependently elevated， and that of cleaved Caspase-3 and Bax was dose-dependently decreased in the YSTLP groups compared with the model group （P<0.01）. Compared with the control group， the mRNA expression levels of CHOP and XBP1 were significantly elevated in the model group （P<0.05， P<0.01）， and the protein expression levels of p-PERK， CHOP， and ATF4 were significantly increased （P<0.05）. Compared with the model group， YSTLP significantly decreased the mRNA expression levels of CHOP and XBP1 （P<0.01） and the protein expression levels of p-PERK， CHOP， and ATF4 （P<0.01）. ConclusionYSTLP can effectively inhibit endoplasmic reticulum stress and improve apoptosis of renal tubular epithelial cells， and its mechanism may be related to the regulation of the PERK/AFT4/CHOP pathway.

Objective:To explore the impact of unidentified injectable fillers on the soft tissue of nasal dorsum and rhinoplasty.Methods:The Plastic Surgery Information System of Xinhua Hospital Affiliated with Dalian University was utilized to conduct an analysis of 62 rhinoplasty patients between 2018 and 2019. Specifically, this included 28 patients with an unidentified history of injectable filler rhinoplasty, encompassing 1 male and 27 females with ages ranging from 19 to 53 years and a mean age of 28.8 years. Additionally, 34 patients underwent primary rhinoplasty, all of whom were female with ages ranging from 19 to 46 years and a mean age of 26.8 years. This study examined the effects of unidentified injectable fillers on the soft tissue of the nasal dorsum by analyzing the excised nasal dorsum under a microscope. Subsequently, statistical methods were performed to assess differences in gender, age, preoperative tip protrusion/nose length, postoperative tip protrusion/nose length, dorsal augmentation modality, and satisfaction, and to investigate the effect of unidentified injectables on the rate of dissatisfaction after rhinoplasty.Results:The histopathological analysis of unidentified injectable fillers removed from the nasal dorsum revealed the presence of mainly gel and granular fillers. The gel fillers, characterized by its pink jelly-like texture, contained unidentified injectable fillers, colorless glue, and were observed to flow out upon cutting. The granular filler, on the other hand, appeared as tough, irregularly shaped tissue similar to caviar. Additionally, evidence of muscle tissue in 5 pathologic sections indicated that the unidentified injectable fillers were injected into or near the dorsal nasal muscles, leading to varying degrees of muscle injuries upon excision. A comparison of 28 rhinoplasty patients with unidentified injectable fillers for nasal dorsal augmentation and 34 patients with primary rhinoplasty showed that 11 females in the former group and 4 females in the latter group were dissatisfied with the results. Statistical analysis demonstrated no significant differences between the two groups in terms of gender ( P=0.452), age ( P=0.219), preoperative tip projection/nasal length ( P=0.681), postoperative tip projection/nasal length ( P=0.105), and nasal dorsum augmentation methods ( P=0.413). However, the initial rhinoplasty group had a lower dissatisfactory rate (4 cases, 11.76%) and the unidentified injectables group had a higher dissatisfactory rate (11 cases, 39.29%), which was statistically significant between the two groups (χ 2=6.341, P=0.012). Conclusions:The presence of unidentified nasal injectable fillers can significantly decrease postoperative satisfactory rates, increase the incidence of dissatisfaction, and have adverse effects on the soft tissues of the nasal dorsum and the overall outcome of the rhinoplasty procedure.

Objective To monitor the susceptibility of clinical isolates to antimicrobial agents in tertiary hospitals in major regions of China in 2022.Methods Clinical isolates from 58 hospitals in China were tested for antimicrobial susceptibility using a unified protocol based on disc diffusion method or automated testing systems.Results were interpreted using the 2022 Clinical &Laboratory Standards Institute(CLSI)breakpoints.Results A total of 318 013 clinical isolates were collected from January 1,2022 to December 31,2022,of which 29.5％were gram-positive and 70.5％were gram-negative.The prevalence of methicillin-resistant strains in Staphylococcus aureus,Staphylococcus epidermidis and other coagulase-negative Staphylococcus species(excluding Staphylococcus pseudintermedius and Staphylococcus schleiferi)was 28.3％,76.7％and 77.9％,respectively.Overall,94.0％of MRSA strains were susceptible to trimethoprim-sulfamethoxazole and 90.8％of MRSE strains were susceptible to rifampicin.No vancomycin-resistant strains were found.Enterococcus faecalis showed significantly lower resistance rates to most antimicrobial agents tested than Enterococcus faecium.A few vancomycin-resistant strains were identified in both E.faecalis and E.faecium.The prevalence of penicillin-susceptible Streptococcus pneumoniae was 94.2％in the isolates from children and 95.7％in the isolates from adults.The resistance rate to carbapenems was lower than 13.1％in most Enterobacterales species except for Klebsiella,21.7％-23.1％of which were resistant to carbapenems.Most Enterobacterales isolates were highly susceptible to tigecycline,colistin and polymyxin B,with resistance rates ranging from 0.1％to 13.3％.The prevalence of meropenem-resistant strains decreased from 23.5％in 2019 to 18.0％in 2022 in Pseudomonas aeruginosa,and decreased from 79.0％in 2019 to 72.5％in 2022 in Acinetobacter baumannii.Conclusions The resistance of clinical isolates to the commonly used antimicrobial agents is still increasing in tertiary hospitals.However,the prevalence of important carbapenem-resistant organisms such as carbapenem-resistant K.pneumoniae,P.aeruginosa,and A.baumannii showed a downward trend in recent years.This finding suggests that the strategy of combining antimicrobial resistance surveillance with multidisciplinary concerted action works well in curbing the spread of resistant bacteria.

Chemical pollution-induced damage to ecosystem function has been a global challenge.The latest"Kunming-Montreal Global Biodiversity Framework"proposed reducing pollution risks to levels harmless to biodiversity and function,placing higher demands on chemical risk management at the ecosystem level.Conventional ecotoxicity tests have focused on single species,only to neglect genetic diversity protection and simplify species interactions.Here,we proposed using environmental RNA(eRNA)and metatranscriptomic analysis to establish a multi-species,multi-biological level chemical pollution ecological risk assessment approach in exposed communities.We reviewed the current status and trends of eRNA in chemical pollution risk assessment and proposed a strategy for bioeffect testing from molecules to communities based on eRNA,constructing ecological risk assessment models for different protection goals.Finally,we summarized the theoretical and technical challenges facing eRNA-based toxicity testing and outlined the future applications of eRNA in capturing real ecological effects of chemical pollution in the field.