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.

Objective:To investigate the effect of neutrophils on cell pyroptosis and its mechanisms in mice with early brain injury (EBI) following subarachnoid hemorrhage (SAH).Methods:Seventy six male C57BL/6J mice were randomly divided into sham-operated group, SAH group, SAH+vehicle group, and SAH+anti-ly6G group ( n=19). SAH models in the latter 3 groups were established by modified endovascular perforation. Mice in the SAH+vehicle group and SAH+anti-ly6G group received intravenous injection of equal normal saline or anti-ly6G antibody (4 mg/kg) 24 h before SAH. At 24 h after SAH, immunofluorescent staining was used to detect the locations/expressions of neutrophils, S100 calcium binding protein A8 (S100A8) and gasdermin D (GSDMD); FJC staining was performed to assess the neuronal injury; modified Garcia test and rotarod test were used to evaluate the neurological functions, and brain water content test was applied to evaluate the brain edema; Western blotting was used to detect the expressions of S100A8, Toll-like receptor 4 (TLR4), NOD-like receptor thermal protein domain associated protein 3 (NLRP3), cleaved cysteinyl aspartate specific proteinase-1 (cleaved-caspase1), and cleaved N-terminal gasdermin D (GSDMD-N). Results:(1) Compared with those in the sham-operated group, neutrophil infiltration at the damaged cortex with highly expressed S100A8 in neutrophils was observed in the SAH group, and increased GSDMD expression at the damaged cortex and GSDMD co-localization in astrocytes, microglia and neurons were observed in the SAH group. (2) Compared with the sham-operated group, the SAH group and SAH+vehicle group had significantly increased numbers of infiltrated neutrophils and FJC-positive neurons, significantly decreased falling latency in the modified Garcia score and rotarod test, significantly increased brain water content, and significantly elevated expressions of S100A8, TLR4, NLRP3, cleaved-caspase1 and GSDMD-N ( P<0.05); the SAH+anti-ly6G group had statistically decreased numbers of infiltrated neutrophils and FJC-positive neurons, statistically increased falling latency in the modified Garcia score and rotarod test, statistically decreased brain water content, and statistically decreased expressions of S100A8, TLR4, NLRP3, cleaved-caspase1 and GSDMD-N compared with the SAH group and SAH+vehicle group ( P<0.05). Conclusion:Inhibition of neutrophils can down-regulate the S100A8 expression after SAH and attenuate TLR4/NLRP3 activation-mediated cell pyroptosis, thereby improving EBI.

Objective:To analyze the clinical characteristics of pertussis cases diagnosed by two pathological detection methods: bacterial culture and real-time polymerase chain reaction (RT-PCR), and to explore the applicable value of two pathological detection methods in the diagnosis of pertussis.Methods:Bilateral nasopharyngeal swabs and clinical information of 165 children suspected of pertussis were collected by Hebei Children′s Hospital from April 2019 to January 2020. The bacterial culture and RT-PCR for nasopharyngeal swab specimens were performed in all cases. Chi-square test was used to analyze the cases of pertussis diagnosed by the above two methods.Results:Based on clinical diagnosis, the sensitivity of bacterial culture and RT-PCR for the diagnosis of pertussis was 61.70% (58/94) and 86.17% (81/94), and the specificity was 92.96% (66/71) and 71.83% (51/71), respectively. The positive rate of RT-PCR in children of all ages, seasons and cough courses is higher than that of bacterial culture. Children with pertussis diagnosed by bacterial culture and RT-PCR were basically similar in age, season, and cough course distribution, with the most common cases ≤3 months old, a high incidence trend in summer and autumn, and the course of coughing in children was mostly within 15-21days. The positive rate of bacterial culture in the diagnosis of pertussis in children is affected by the age of the children, and there are statistical differences between children in different age groups (χ2= 11.929, P=0.036). The positive rate of bacterial culture was the highest in children with >3 years old (51.85% [14/27]), followed by children with ≤3 months old (48.72% [19/39]), and the lowest in children with >6-12 months old (15.00% [3/20]). Moreover, the positive rate of bacterial culture in the diagnosis of pertussis in children is also affected by the cough course of the children, and there are statistical differences between children in different cough course groups (χ2=9.841, P=0.020). The positive rate of bacterial culture was the highest in children with cough course 15-21 days (49.23% [32/65]), followed by 43.59% (17/39) in children with cough course 8-14 days, and the lowest in children with cough course of less than 7 days (22.86% [8/35]). Conclusions:Compared with RT-PCR, bacterial culture has lower sensitivity and higher specificity in the detection of pertussis. These two detection methods have their own advantages and limitations. Medical institutions at all levels should comprehensively analyze different laboratory detection methods. Only by combining the two methods can the diagnostic value and level be effectively improved.

The TEA domain (TEAD) family proteins (TEAD1‒4) are essential transcription factors that control cell differentiation and organ size in the Hippo pathway. Although the sequences and structures of TEAD family proteins are highly conserved, each TEAD isoform has unique physiological and pathological functions. Therefore, the development and discovery of subtype selective inhibitors for TEAD protein will provide important chemical probes for the TEAD-related function studies in development and diseases. Here, we identified a novel TEAD1/3 covalent inhibitor (DC-TEADin1072) with biochemical IC

Objective:To investigate frailty status of the elderly patients with leukoaraiosis (LA) and to analyze the correlation between white matter hyperintensities and their frailty status.Methods:From June 2019 to September 2020, a total of 162 patients with leukoaraiosis over 65 years old were screened by cranial MRI.The Fried frailty phenotype was used to evaluate their frailty status.The Fazekas scale scoring method was used to independently assess the periventricular white matter hyperintense (PVH) and deep white matter hyperintense (DWMH) by the cranial MRI images.SPSS 22.0 software was used for statistical analysis.ANOVA test was used to compare normal distribution data between groups, and Kruskal-Wallis H test was used to compare non-normal distribution data between groups.Spearman rank correlation analysis was used to analyze the correlation between PVH and DWMH scores and Fried frailty phenotype score. Results:Among 162 elderly patients with leukoaraiosis, 46 patients (28.40%) were non-frailty, 76 patients (46.91%)were pre-frailty and 40 patients (24.69%) were frailty.There were statistically significant differences in age( F=9.382, P<0.01), number of chronic diseases( H=10.736, P<0.01), number of medication ( H=15.927, P<0.01) and mini-nutritional assessment short form (MNA-SF) scores( F=5.263, P<0.01) among older LA patients with different frailty phenotype.There was statistical difference in PVH scores in elderly LA patients with different frailty phenotype (χ 2=108.537, P<0.01), but no significant difference in DWMH scores (χ 2=4.239, P>0.05). Spearman correlation analysis showed significant positive correlation between PVH score and frailty phenotype score in elderly LA patients ( r=0.718, P<0.001). Conclusion:Elderly LA patients have a high incidence of frailty, which may be related to aging, multi-disease coexistence, multiple medication, nutritional risk and other factors.The occurrence of weakness in elderly LA patients is related to periventricular white matter lesions, and the more serious the white matter damage, the more obvious the degree of frailty.

Objective:To analyze the gait and balance disorder and fall risk in patients with leukoaraiosis (LA) in different degrees and locations.Methods:Seventy-eight patients with LA (LA group) and thirty healthy controls (NC group) were selected.According to Fazekas grading scale, the LA group were subgroups of different degrees(LA1, LA2, LA3)and evaluated independent scores for periventricular white matter hypersignal (PVH) and deep white matter hypersignal (DWMH). All subjects were tested by Tinetti scale, and the gait test, balance test and fall risk assessment were carried out, and the correlation between gait test results and balance test results and PVH and DWMH scores were analyzed.Results:There were significant differences in gait test((11.47±0.51), (10.18±1.29)), balance test((15.63±0.49), (13.96±1.58)) and fall risk((27.10±0.76), (24.15±2.73)) between LA group and NC group ( t=5.310, 5.667, 5.799, all P< 0.01). There were no significant differences in gait test, balance test and fall risk between NC group and LA1 group (all P>0.05). There were significant differences in gait test, balance test and fall risk between NC group and LA2 group and LA3 group ( t=1.110, 1.562, 1.336; 2.592, 3.342, 2.985; all P<0.01). There were significant differences in gait test, balance test and fall risk between LA1 group((11.19±0.85), (15.38±0.75), (26.58±1.47)) and LA2 group((10.36±0.82), (14.07±1.09), (24.43±1.64)) and LA3 group((8.88±0.99), (12.23±1.08), (21.21±1.93)) ( t=0.835, 1.313, 1.039; 2.317, 3.093, 2.756; all P<0.01). There were significant differences in gait test, balance test and fall risk between LA2 group and LA3 group ( t=1.482, 1.780, 1.639, all P<0.01). The analysis showed that the gait test was negatively correlated with the PVH and DWMH scores ( r=-0.810, P<0.01; r=-0.452, P<0.01). The balance test was negatively correlated with the PVH and DWMH scores ( r=-0.502, P<0.01; r=-0.836, P<0.01). Conclusions:There are gait disorder and balance disorder in patients with moderate and severe LA.With the increase of the degree of LA , the damage of gait and balance function is significantly increased, and the risk of fall is also significantly increased.The gait disorder and the balance disorder of LA patients has different correlation with the high-signal score of the white matter in different parts.The correlation between gait disorder and PVH score is more obvious, and the correlation between balance disorder and DWMH score is more obvious.

Objective: By detecting the expression of interleukin-13 (IL-13) and periostin in the airway of asthmatic patients, the pathological changes and pulmonary functions of airway tissues in asthmatic patients were evaluated, and the role of IL-13 and periostin airway remodeling in bronchial asthma was preliminarily explored. Methods: The bronchial tissues adjacent to tumor nest were obtained from 12 patients with lung cancer complicated with bronchial asthma (asthmatic group) and 12 lung cancer patients without bronchial asthma (non-asthmatic group) after lung cancer resection. Pulmonary function was measured for all subjects before surgery. Pathological changes of airway tissues and degree of airway remodeling were assessed by hematoxylin-eosin (H&E) staining, masson′s trichrome staining, and periodic acid-silver methenamine (PASM) staining of paraffin-embedded sections. The expression of IL-13 and periostin in bronchial tissues were evaluated by immunohistochemistry. Results: Values of the forced expiratory volume in 1 second of the predicted value (FEV1% pred) and FEV1/forced vital capacity (FEV1/FVC%) in asthmatic patients were significantly decreased compared with the non-asthmatic patients (P<0.05), indicating that lung function was impaired in asthmatic patients. There was more severe airway remodeling representing as thickening of basement membranes, collagen deposition, and increasing of goblet cells and fibroblasts in asthmatic patients than in non-asthmatic patients (all P<0.05). The expression of IL-13 and periostin were higher in asthmatic tissues than in non-asthmatic tissues (P<0.05). The immunohistochemical expression of IL-13 and periostin in bronchial tissues were positively correlated with the degree of airway remodeling in asthmatic patients, and the expression of IL-13 and periostin in bronchial tissues were positively correlated with each other. Conclusions The expression of IL-13 and periostin were increased in bronchial tissue in patients with asthma. They work together to promote the occurrence of airway remodeling, which eventually lead to a decline in lung function.

Objective To investigate the role of hesperitin in regulating inflammatory response in early brain injury after subarachnoid hemorrhage (SAH). Methods A total of 96 adult male SD rats were divided into sham-operated group, SAH group, solvent group and intervention group (n=24) by random number table method. The SAH rat models in the latter three groups were prepared by carotid artery puncture method; the rats in the intervention group were given oral administration of hesperidin solution, which was dissolved in 5％ dimethyl sulfolide (DMSO), with a concentration of 1 mg/100 μL and a dosage of 40 mg/kg within 30 min after operation; rats in the solvent group were given oral administration of an equal volume of 5％ DMSO solution. Modified Garcia behavioral scale was used to evaluate the neurobehavior of rats, and the wet/dry weight method was used to measure the water content in the brain tissues of the left and right hemispheres of the rats 24 h after SAH. Immunofluorescence staining was used to detect the microglia activation, Fluoro-dyed Jade C staining was used to assess the brain neuron degeneration, enzyme-linked immunosorbent assay (ELISA) was employed to detect the inflammatory factors interleukin (IL)-1β, IL-6 and tumor necrosis factor-α (TNF-α) content in the brain tissues, and Western blotting was used to detect the nuclear factor-κB (NF-κB) and phosphorylated (p) -NF-κB protein expressions. Results As compared with the solvent group, intervention group had significantly increased improved modified Garcia behavioral scale scores (10.08±1.73 vs. 13.83±1.70), and significantly decreased water content of brain tissues in the left and right hemispheres ([81.44 ± 1.05]％ vs. ([79.14±0.82]％; [80.55±1.55]％ vs. [78.79±1.02]％), significantly smaller number of CD68+ and Iba1+ microglias (30.17±1.04 vs. 10.67±0.75; 29.33±1.16 vs. 12.00±0.41), significantly smaller number of degenerate neurons (53.21±0.94 vs. 31.33±0.28), significantly reduced levels of inflammatory cytokines IL-1β, IL-6 and TNF-α ([429.88±106.32] pg/mL vs. [221.50±48.80] pg/mL; [1015.50±221.80] pg/mL vs. [448.11±93.40] pg/mL; [1021.75±149.17] pg/mL vs. [595.71±190.81] pg/mL), and significantly lower p-NF-κB/NF-κB ratio (1.13±0.07 vs. 0.71±0.02, P<0.05). Conclusion Hesperitin may reduce the inflammatory response mediated by microglia after subarachnoid hemorrhage by inhibiting NF-κB pathway, thereby improving the neurological dysfunction of rats.

Objective To evaluate the effect of cerebral microbleeds (CMBs) on intracranial aneurysm rupture and hemorrhage. Methods A total of 2023 patients with intracranial single aneurysms (944 un-ruptured aneurysms and 1029 ruptured aneurysms) were enrolled in our study. The 3D-DSA was applied to all patients to evaluate the aneurysm sizes, locations, and morphous features, and to confirm the presence of A1 dominance and variations of Circle of Willis; moreover, aspect ratio (AR), size ratio (SR) and aneurismal inclination angle were measured. The presence of CMBs identified by T2-weighted gradient-recalled-echo sequence on magnetic resonance imaging (MRI) was evaluated; the clinical data of combined with/without CMBs patients were compared. Ninety-two untreated intracranial aneurysms patients combined with CMBs were followed up to evaluate the incubation periods of CMBs-related intracranial aneurysm rupture and hemorrhage; the clinical data of patients with un-ruptured aneurysms and ruptured aneurysms were compared; the risk factors of intracranial aneurysm rupture and hemorrhage were analyzed by univariate and multivariate Logistic regression analyses. Results CMBs confirmed by MR imaging were presented in 158 patients, with 7.81％ incidence rate (158/2023). Age, proportion of smokers, aneurysm inclination Angle, SR, narrow neck, irregular aneurysm shape, proportion of rupture and hemorrhage, aneurysm sites, and hypertension showed significant differences between patients without CMBs and patients with CMBs (P<0.05). In the untreated intracranial aneurysms patients combined with CMBs, 27 had intracranial aneurysm rupture and 65 did not appear intracranial aneurysm rupture; the rupture time was 3-46 months, with an average of (15.07± 10.76) months. As compared with the un-ruptured group, the ruptured group had a statistically higher proportion of patients with irregular aneurysm morphology (P<0.05). Univariate analysis showed that CMBs, female, age, aneurysm size, aneurysm morphology, ICA and ACA aneurysms, AR, variations of Circle of Willis, hypertension grading II and III, diabetes mellitus with fasting blood glucose≤6.0 mmol/L, hyperlipidemia, coronary heart disease, and drinking alcohol were important factors affecting intracranial aneurysm rupture, and the differences were statistically significant (P<0.05). Multivariate Logistic regression analysis showed that CMB was an independent risk factor for intracranial aneurysm rupture and hemorrhage; as compared with patients without CMBs, patients with CMBs had a 1.75 fold increased risk of aneurysm rupture. Conclusions Patients with intracranial aneurysms with older age, smaller aneurysm inclination Angle and larger SR are more likely to be associated with CMBs. Intracranial aneurysms with CMBs patients with irregular morphology are prone to have rupture and hemorrhage. CMBs is an independent risk factor for intracranial aneurysm rupture and hemorrhage.

Objective: To investigate the effect of DAMP (damaged associated molecular pattern) on the inhibition of RAS-mutant A549 lung adenocarcinoma cells by CIK cells and its mechanism. Methods: Human peripheral blood mononuclear cells were isolated in vitro and CIK cells were cultured. A549 cells were treated with cisplatin (DDP) and doxorubicin (ADM) alone or in combination, and the morphology of A549 cells was observed under a microscope. The supernatant of A549 cells was co-cultured with CIK cells. Flow cytometry was used to detect the CIK cell immunophenotype after co-culture. MTT assay was used to detect the inhibition of A549 lung cancer cell proliferation induced by A549 cell supernatant. The concentration of chemotherapeutic drugs kills A549 cell supernatant CRT, ATP, HMGB1 content. Results: Low-level chemotherapeutic drugs showed more immunogenic death characteristics after killingA549 cells. The ratio of CD8+ and CD56+ in CIK cells was significantly higher than that in control CIK cells (P<0.05). The inhibition rate of CIK cells induced byA549 cells after injury onA549 lung adenocarcinoma cells was significantly higher than that of the same dose chemotherapy group [DDP group (31.34±1.51)% vs (5.97±1.74)%, ADM group (45.46±1.78)% vs (6.22±1.34)%, DDP+ ADM group (45.78±1.14)% vs (11.94±3.11)%, all P<0.05], and low-mass chemotherapeutic agents killed C549 induced by A549 cell supernatant on A549 The inhibition rate of the cells was higherthan that of the supernatant induced by the higher concentration of chemotherapeutic drugs (all P<0.05). The level of CRT,ATP, and HMGB1 in immunogenicity-related molecules in the supernatant ofA549 cells was significantly increased by low-concentration chemotherapy drugs (all P<0.05). In the low-concentration group, the supernatant-induced inhibition of the proliferation of A549 lung adenocarcinoma cells increased with the increase of CRT, ATP, and HMGB1 levels. Conclusion: The combination of lower concentration of DDP and ADM alone or in combination could more easily induce the immunogenic death of A549 cells and release higher levels of DAMP molecules, which could promote the inhibitory effect of CIK on lung cancerA549 cells.