Objective : To observe the brain tissue injury during hypoxia-ischemia, as well as the pathological changes and the expression of ferroptosis-related factors after the use of Shenfu injection(SFI), and to explore the protective effect of SFI on hypoxic-ischemic brain injury(HIBD) by inhibiting ferroptosis. Methods : An animal model of HIBD in SD rats was constructed and intervened with SFI. Pathologic changes in brain tissue were observed by HE staining methods. Nissen staining was used to observe neuron survival. Glutathione Peroxidase 4(GPX4) and Divalent Metal Transporter 1(DMT1) expression were detected in brain tissue by Western blot, immunohistochemistry and immunofluorescence. Reduced Glutathione(GSH), Lactate Dehydrogenase(LDH), Malondialdehyde(MDA), Superoxide Dismutase(SOD) and tissue iron content were determined with the kits. BV-2 microglial cell line(BV2) cells were culturedin vitroand divided into control group(Ctrl group), oxygen-glucose deprivation group(OGD group), iron ferroptosis-inducing group(Erastin group), iron ferroptosis-inhibiting group(Fer-1 group), Shenfu injection group(SFI group), and Erastin+Shenfu injection group(Erastin+SFI group). 2′,7′-Dichlorodihydrofluorescein diacetate(DCFH-DA) reactive oxygen species(ROS) fluorescent probe was used to detect the ROS release level; Immunofluorescence was used to observe intracellular GPX4, DMT1 expression. Results : Compared with the Sham group, rats in the HIBD group showed significant neuronal cell damage in brain tissue, decreased GPX4 expression(P<0.01), increased DMT1 expression(P<0.01), decreased GSH and SOD levels(P<0.01), and increased LDH, MDA and tissue iron levels(P<0.05,P<0.05,P<0.01). In contrast, after the intervention of SFI, GPX4 expression was elevated(P<0.01), DMT1 expression decreased(P<0.01), GSH and SOD levels were elevated(P<0.01), and LDH, MDA, and tissue iron levels decreased(P<0.05,P<0.05,P<0.01). The cells experiments showed that compared with the Ctrl group, the OGD group had a significantly higher ROS content and a decrease in the expression of GPX4 fluorescence intensity, and an increase in the fluorescence intensity of DMT1(P<0.01), compared with the OGD group, the ROS content was reduced in the SFI group, while the expression of GPX4 was elevated and the expression of DMT1 was reduced(P<0.01). Conclusion Hippocampal and cortical regions are severely damaged after HIBD in neonatal rats, and their brain tissues show decreased expression of GPX4 and increased expression of DMT1. The above suggests that ferroptosis is involved in HIBD brain injury in neonatal rats. In contrast, Shenfu injection has a protective effect on HIBD experimental animal model and BV2 cell injury model by reducing iron aggregation and ROS production.

OBJECTIVE To establish a clinical comprehensive evaluation framework for direct oral anticoagulants （DOACs） in the prevention of cancer-associated venous thromboembolism （CAVTE）， providing a methodological reference for the rational prevention and treatment of CAVTE as well as for the formulation and adjustment of macro-management strategies for anticoagulant drugs. METHODS Through literature retrieval， evaluation indicators were collected and organized to establish a preliminary indicator pool. The selection of evaluation indicators was carried out through two rounds of Delphi surveys using average score of indicator importance≥3.5 and a coefficient of variation （CV） ＜0.25 as the screening criteria. Analytic hierarchy process （AHP） was employed to finalize the indicator weights. RESULTS The authority levels （C）r of the two rounds of expert consultations were 0.877 and 0.943， with CV of 0.24 and 0.18， respectively. The Kendall concordance coefficients were 0.331 and 0.535 （P＜0.05）. After expert validation， six primary indicators and forty-six secondary indicators were finalized for inclusion in the evaluation framework. The primary indicators and their weightings， ranked in descending order， were as follows：“ effectiveness” （38.86%）， “safety” （38.86%），“ cost-effectiveness” （10.67%），“ accessibility” （5.51%），“ suitability” （3.48%）， and “innovation” （2.64%）. The secondary indicators exhibited a weight range from 0.02% to 20.25%， with the top five secondary indicators being：“ incidence of intracranial hemorrhage” （20.25%）， “reduction in all-cause mortality” （15.29%）， “decrease in the incidence of pulmonary embolism” （8.82%）， “reduction in the incidence of deep vein thrombosis” （7.25%）， and “drug contraindications” （4.74%）. CONCLUSIONS This study has established an authoritative， scientific， and reliable comprehensive clinical evaluation framework for the use of DOACs in the prevention of CAVTE.

ObjectiveTo analyze the genomic characteristics of Streptococcus pyogenes (GAS) isolated from children with respiratory tract infections in a tertiary hospital in Jinshan District of Shanghai during 2013‒2024, to compare the changes in trend for genomic characteristics before and after 2000, and to provide scientific data for the prevention and control of GAS infections. MethodsGAS strains isolated from children with respiratory tract infections in this hospital were collected from 2013 to 2024. Antimicrobial susceptibility of the isolated strains to 12 antibiotics, including penicillin, cefotaxime, cefepime, linezolid, vancomycin, meropenem, chloramphenicol, ofloxacin, levofloxacin, erythromycin, clindamycin, and tetracycline, was determined using broth microdilution plate method. Besides, whole genome sequencing (WGS) was used to analyze multilocus sequence type (MLST), emm typing, carriage of superantigen genes, mobile genetic element (MGE), carriage of virulence gene, and genomic phylogenetic tree of the isolated strains. ResultsA total of 50 GAS strains were collected and identified from children with respiratory tract infections aged 4‒14 years old, and the resistance rates of those isolates to erythromycin, clindamycin, and tetracycline were 100.00%, 100.00%, and 86.00%, respectively. There were two emm types in the GAS isolates; the emm12 type accounted for 76.00% (38/50), corresponding to ST36 type, and the emm1 type accounted for 24.00% (12/50), corresponding to ST28, ST1274, and new-1 types. There was a statistically significant difference in the constitution of the MLST before and after 2020 (P=0.015). All the isolates carried the superantigen genes speC, speG, ssa, and smeZ. The predominant emm12 isolates belonged to the Clade Ⅱ, carrying the mobile elements ICE-emm12 (harboring erythromycin-resistance gene ermB and tetracycline-resistance gene tetM) and ΦHKU.vir (carrying virulence genes speC and ssa). The emm1 isolates carried the mobile elements ICE-HKU488 (harboring erythromycin-resistance gene ermB and tetracycline-resistance gene tetM) and ΦHKU488.vir (carrying virulence genes speC and ssa), and had close phylogenetical relationships with isolates from Hong Kong, China. No M1_UK new clone strains were found. The ST1274 isolates of emm1 were newly discovered in 2020‒2024, and belonged to a separate phylogenetic clade. ConclusionGAS strains isolated from children with respiratory tract infections in a tertiary hospital in Jinshan District of Shanghai exhibit a high resistance to erythromycin, clindamycin, and tetracycline. It is recommended that the clinical treatments change to use other antimicrobial drugs, such as penicillin, third-generation cephalosporins, and fluoroquinolones. During 2020‒2024, a new ST1274 clone strain is discovered in emm1 GAS isolates, without M1_UK new clone strains being found. It is essential to continuously concern locally prevalent GAS strains and perform early identification of MLST types to promptly monitor the internal changes of the bacterial population and potential prevalence of new clones.

[摘 要] 目的：探究芒柄花素调节JAK2/STAT3信号通路对胆囊癌细胞GBC-SD恶性生物学行为及血管生成的影响。方法：常规培养GBC-SD细胞并构建其移植瘤裸鼠，将GBC-SD细胞和其移植瘤裸鼠分为对照组、芒柄花素组、colivelin（JAK2/STAT3激活剂）组、芒柄花素 + colivelin组。用MTT法、Transwell实验和流式细胞术分别检测各组细胞的增殖、迁移和侵袭能力，以及凋亡，用ELISA法检测各组细胞血管内皮生长因子（VEGF）分泌水平，血管生成拟态（VM）形成实验检测各组细胞的成管腔能力，移植瘤实验检测芒柄花素对移植瘤生长的影响，免疫组化法检测各组移植瘤组织中CD31和VEGF表达，WB法检测各组细胞和移植瘤组织中JAK2/STAT3通路的磷酸化水平。结果：芒柄花素可明显抑制GBC-SD细胞的增殖、迁移和侵袭能力，促进其凋亡（均P < 0.05），抑制GBC-SD细胞分泌VEGF和VM形成（均P < 0.05），抑制GBC-SD细胞移植瘤的生长及血管生成（均P < 0.05），抑制移植瘤组织中VEGF表达，抑制GBC-SD细胞和其移植瘤组织中JAK2/STAT3通路的磷酸化，colivelin均可逆转芒柄花素的上述作用（均P < 0.05）。结论：芒柄花素通过抑制JAK2/STAT3信号通路抑制GBC-SD细胞增殖、迁移、侵袭、血管形成，促进其凋亡，JAK2/STAT3信号通路是胆囊癌临床治疗的潜在靶点。

Objective:To explore the expression of lncRNA ARHGAP5-AS1 in renal cancer tissues and cell lines, and the effect of ARHGAP5-AS1 on the proliferation and invasion of renal cancer cell lines and its molecular mechanism.Methods:The GEPIA database was used to analyze the expression of ARHGAP5-AS1 in renal cancer tissues, and its relationship with clinical stage, overall survival and disease-free survival of renal cancer patients was analyzed. Real-time fluorescence quantitative polymerase chain reaction (RT-qPCR) was used to detect the expression level of ARHGAP5-AS1 in renal cancer cells (786-O, Caki-1, OS-RC-2, ACHN, A-498). Renal carcinoma OS-RC-2 cells were transfected with pcDNA3.1-ARHGAP5-AS1 plasmid or pcDNA3.1 plasmid, denoted as ARHGAP5-AS1 group and control group. Colony formation assay and Transwell assay were used to detect changes in the proliferation and invasion ability of OS-RC-2 cells. Dual-luciferase reporter gene experiment was used to verify the targeting relationship between ARHGAP5-AS1 and miR-155-5p. The Starbase v3.0 online database was used to analyze the correlation between the expression of ARHGAP5-AS1 and miR-155-5p in renal cancer tissues. RT-qPCR was used to detect the expression level changes of miR-155-5p. Western blotting was used to detect the expression changes of Raf/MEK/ERK molecular pathway proteins p-Raf, p-MEK, p-ERK, p-FBW7, and c-MYC. The measurement data were expressed as mean ± standard deviation ( ± s), the independent sample t-test was used for comparison between two groups, and one-way analysis of variance was used for comparison between multiple groups. Results:ARHGAP5-AS1 was lowly expressed in renal cancer tissues ( P<0.01), and its expression level was related to the clinical stage, overall survival and disease-free survival of patients with renal cancer ( P<0.01). ARHGAP5-AS1 showed low expression in renal cancer cell lines (786-O, Caki-1, OS-RC-2, ACHN, A-498) ( P<0.01). Compared with the control group, the proliferation and invasion abilities of OS-RC-2 cells in ARHGAP5-AS1 group were significantly reduced ( P<0.01). Dual-luciferase reporter gene experiment confirmed that ARHGAP5-AS1 targets and binds to miR-155-5p ( P<0.01). The expression of ARHGAP5-AS1 and miR-155-5p in renal cancer tissues was negatively correlated ( P<0.01). Compared with the control group, the expression of miR-155-5p in OS-RC-2 cells in the ARHGAP5-AS1 group was significantly reduced ( P<0.01). Compared with the control group, the expression levels of Raf/MEK/ERK molecular pathway proteins p-Raf, p-MEK, p-ERK, p-FBW7, and c-MYC in OS-RC-2 cells in the ARHGAP5-AS1 group were reduced. Conclusions:lncRNA ARHGAP5-AS1 is lowly expressed in renal cancer tissues and is related to the clinical stage and survival of renal cancer patients. ARHGAP5-AS1 inhibits the proliferation and invasion of renal cancer cells by targeting the expression of miR-155-5p.

Objective To compare the efficacy of blood letting under pain point touch and ultrasound-guided puncture de-compression in the treatment of acute supraspinatus muscle calcifying tendinitis.Methods From January 2020 to January 2023,45 patients with acute supraspinatus muscle calcifying tendinitis were selected and divided into treatment group and control group.In the treatment group,a total of 22 patients were treated with ultrasound-guided puncture decompression,in-cluding 16 females and 6 males,aged from 20 to 64 years old(39.31±5.80)years old,11 on the left shoulder and 11 on the right shoulder.In the control group,there were 23 cases,including 15 females and 8 males,aged from 19 to 66 years old(40.67±6.13)years old,12 on the left shoulder and 13 on the right shoulder.The treatment was treated with pain point touch bloodletting therapy.The visual analog scale(VAS)pain score,University of California,Los Angeles(UCLA)shoulder system score and shoulder Constant-Murley score were used to evaluate the therapeutic effect before treatment,1 weeks,1 month and 3 months after treatment,respectively.Results One patient in the control group gave up follow-up for personal reasons after 1 week of treatment,and the other 44 patients completed all follow-up.Six months after treatment,there were no recurrence cases in both groups.After statistical analysis,VAS pain score,UCLA score and Constant-Murley score of the treatment group and the control group were significantly different from those before treatment(P＜0.05),and the improvement was more obvious in the treatment group.There was no statistical significance between the two groups(P＞0.05).Conclusion Bloodletting under pain point touch and ultrasound-guided puncture decompression are effective in the treatment of acute calcific supraspinatus tendinitis,with simple operation and low cost,which can effectively reduce local pain and effectively improve shoulder joint function.Primary hospitals can selectively operate treatment according to their own conditions.

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 distribution and antimicrobial resistance profiles of the common pathogens isolated from urine from 2015 to 2021 in the CHINET Antimicrobial Resistance Surveillance Program.Methods The bacterial strains were isolated from urine and identified routinely in 51 hospitals across China in the CHINET Antimicrobial Resistance Surveillance Program from 2015 to 2021.Antimicrobial susceptibility was determined by Kirby-Bauer method,automatic microbiological analysis system and E-test according to the unified protocol.Results A total of 261 893 nonduplicate strains were isolated from urine specimen from 2015 to 2021,of which gram-positive bacteria accounted for 23.8％(62 219/261 893),and gram-negative bacteria 76.2％(199 674/261 893).The most common species were E.coli(46.7％),E.faecium(10.4％),K.pneumoniae(9.8％),E.faecalis(8.7％),P.mirabilis(3.5％),P.aeruginosa(3.4％),SS.agalactiae(2.6％),and E.cloacae(2.1％).The strains were more frequently isolated from inpatients versus outpatients and emergency patients,from females versus males,and from adults versus children.The prevalence of ESBLs-producing strains in E.coli,K.pneumoniae and P.mirabilis was 53.2％,52.8％and 37.0％,respectively.The prevalence of carbapenem-resistant strains in E.coli,K.pneumoniae,P.aeruginosa and A.baumannii was 1.7％,18.5％,16.4％,and 40.3％,respectively.Lower than 10％of the E.faecalis isolates were resistant to ampicillin,nitrofurantoin,linezolid,vancomycin,teicoplanin and fosfomycin.More than 90％of the E.faecium isolates were ressitant to ampicillin,levofloxacin and erythromycin.The percentage of strains resistant to vancomycin,linezolid or teicoplanin was＜2％.The E.coli,K.pneumoniae,P.aeruginosa and A.baumannii strains isolated from ICU inpatients showed significantly higher resistance rates than the corresponding strains isolated from outpatients and non-ICU inpatients.Conclusions E.coli,Enterococcus and K.pneumoniae are the most common pathogens in urinary tract infection.The bacterial species and antimicrobial resistance of urinary isolates vary with different populations.More attention should be paid to antimicrobial resistance surveillance and reduce the irrational use of antimicrobial agents.

Objective To understand the distribution and changing resistance profiles of clinical isolates of Enterococcus in hospitals across China from 2015 to 2021.Methods Antimicrobial susceptibility testing was conducted for the clinical isolates of Enterococcus according to the unified protocol of CHINET program by automated systems,Kirby-Bauer method,or E-test strip.The results were interpreted according to the Clinical & Laboratory Standards Institute(CLSI)breakpoints in 2021.WHONET 5.6 software was used for statistical analysis.Results A total of 124 565 strains of Enterococcus were isolated during the 7-year period,mainly including Enterococcus faecalis(50.7％)and Enterococcus faecalis(41.5％).The strains were mainly isolated from urinary tract specimens(46.9％±2.6％),and primarily from the patients in the department of internal medicine,surgery and ICU.E.faecium and E.faecalis strains showed low level resistance rate to vancomycin,teicoplanin and linezolid(≤3.6％).The prevalence of vancomycin-resistant E.faecalis and E.faecium was 0.1％and 1.3％,respectively.The prevalence of linezolid-resistant E.faecalis increased from 0.7％in 2015 to 3.4％in 2021,while the prevalence of linezolid-resistant E.faecium was 0.3％.Conclusions The clinical isolates of Enterococcus were still highly susceptible to vancomycin,teicoplanin,and linezolid,evidenced by a low resistance rate.However,the prevalence of linezolid-resistant E.faecalis was increasing during the 7-year period.It is necessary to strengthen antimicrobial resistance surveillance to effectively identify the emergence of antibiotic-resistant bacteria and curb the spread of resistant pathogens.

Objective To examine the changing antimicrobial resistance profile of Enterobacter spp.isolates in 53 hospitals across China from 2015 t0 2021.Methods The clinical isolates of Enterobacter spp.were collected from 53 hospitals across China during 2015-2021 and tested for antimicrobial susceptibility using Kirby-Bauer method or automated testing systems according to the CHINET unified protocol.The results were interpreted according to the breakpoints issued by the Clinical & Laboratory Standards Institute(CLSI)in 2021(M100 31st edition)and analyzed with WHONET 5.6 software.Results A total of 37 966 Enterobacter strains were isolated from 2015 to 2021.The proportion of Enterobacter isolates among all clinical isolates showed a fluctuating trend over the 7-year period,overall 2.5％in all clinical isolates amd 5.7％in Enterobacterale strains.The most frequently isolated Enterobacter species was Enterobacter cloacae,accounting for 93.7％(35 571/37 966).The strains were mainly isolated from respiratory specimens(44.4±4.6)％,followed by secretions/pus(16.4±2.3)％and urine(16.0±0.9)％.The strains from respiratory samples decreased slightly,while those from sterile body fluids increased over the 7-year period.The Enterobacter strains were mainly isolated from inpatients(92.9％),and only(7.1±0.8)％of the strains were isolated from outpatients and emergency patients.The patients in surgical wards contributed the highest number of isolates(24.4±2.9)％compared to the inpatients in any other departement.Overall,≤ 7.9％of the E.cloacae strains were resistant to amikacin,tigecycline,polymyxin B,imipenem or meropenem,while ≤5.6％of the Enterobacter asburiae strains were resistant to these antimicrobial agents.E.asburiae showed higher resistance rate to polymyxin B than E.cloacae(19.7％vs 3.9％).Overall,≤8.1％of the Enterobacter gergoviae strains were resistant to tigecycline,amikacin,meropenem,or imipenem,while 10.5％of these strains were resistant to polycolistin B.The overall prevalence of carbapenem-resistant Enterobacter was 10.0％over the 7-year period,but showing an upward trend.The resistance profiles of Enterobacter isolates varied with the department from which they were isolated and whether the patient is an adult or a child.The prevalence of carbapenem-resistant E.cloacae was the highest in the E.cloacae isolates from ICU patients.Conclusions The results of the CHINET Antimicrobial Resistance Surveillance Program indicate that the proportion of Enterobacter strains in all clinical isolates fluctuates slightly over the 7-year period from 2015 to 2021.The Enterobacter strains showed increasing resistance to multiple antimicrobial drugs,especially carbapenems over the 7-year period.