ObjectiveTo investigate the effects of Tianma Goutengyin on the tumor necrosis factor-α （TNF-α）/signal transducer and activator of transcription 3 （STAT3）/α1-antichymotrypsin C-terminal tail fragment （α1ACT） signaling pathway and A1-type astrocytes in a rat model of vascular dementia. MethodsSeventy-two male Sprague-Dawley rats were randomly divided into six groups （n=12 per group）： Sham-operated group， model group， Tianma Goutengyin high-， medium-， and low-dose groups （5.13， 10.26， and 20.52 g·kg^-1）， and a nimodipine group （8.1 mg·kg^-1）. The vascular dementia model was established by permanent bilateral common carotid artery occlusion， followed by 4 weeks of intervention. Learning and memory ability were evaluated using the novel object recognition test， and behavioral performance was assessed using the forced swimming test. Levels of interleukin-6 （IL-6） and C-C motif chemokine ligand 2 （CCL2） in hippocampal tissue were measured by enzyme-linked immunosorbent assay （ELISA）. Hippocampal neuronal morphology was observed by Nissl staining， and apoptosis was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling （TUNEL）. Immunohistochemistry was used to detect positive expression of brain-derived neurotrophic factor （BDNF）， glial fibrillary acidic protein （GFAP）， and myelin basic protein （MBP）. Western blot analysis was performed to measure the protein expression levels of TNF-α， TNF receptor 1 （TNFR1）， phosphorylated STAT3 （p-STAT3）， α1ACT， IL-6， complement component 3 （C3）， BDNF， S100 calcium-binding protein A10 （S100A10）， and GFAP in hippocampal tissue. ResultsCompared with the sham-operated group， the model group showed a significantly reduced relative recognition index in the novel object recognition test （P<0.01）， prolonged immobility time and increased immobility frequency in the forced swimming test （P<0.01）. Hippocampal IL-6 and CCL2 levels were significantly increased （P<0.01）. Nissl staining revealed a marked reduction in neuronal number and loss of Nissl bodies （P<0.01）. MBP-positive expression was significantly decreased （P<0.01）， apoptosis was significantly increased （P<0.01）， BDNF-positive expression was significantly reduced （P<0.05）， and GFAP-positive expression was significantly increased （P<0.01）. In addition， the protein expression levels of TNF-α， TNFR1， p-STAT3， α1ACT， IL-6， and C3 were significantly elevated （P<0.01）， while BDNF and S100A10 expression levels were significantly decreased （P<0.01）. Compared with the model group， all Tianma Gouteng yin dose groups exhibited a significant increase in the relative recognition index （P<0.05）， shortened immobility time and reduced immobility frequency （P<0.05， P<0.01）. IL-6 and CCL2 levels were significantly decreased （P<0.01）， neuronal number was significantly increased （P<0.05， P<0.01）， and MBP-positive expression was significantly enhanced （P<0.01）. Apoptosis was significantly reduced （P<0.01）， BDNF-positive expression was significantly increased （P<0.05）， and GFAP-positive expression was significantly decreased （P<0.01）. Moreover， the protein expression levels of TNF-α， TNFR1， p-STAT3， α1ACT， IL-6， and C3 were significantly decreased （P<0.01）， while BDNF and S100A10 protein expression levels were significantly increased （P<0.01）. ConclusionTianma Goutengyin may inhibit A1-type astrocyte activation in rats with vascular dementia through the TNF-α/STAT3/α1ACT signaling pathway， thereby reducing neuronal apoptosis and improving learning and memory function.

ObjectiveTo evaluate the detection specificity for clinical samples and the detection capability for standard substances of five commercially available multiplex polymerase chain reaction (PCR) nucleic acid detection kits (hereinafter referred to as the kits) for respiratory pathogens, and to provide a reference for selecting appropriate detection kits for multi-pathogen nucleic acid testing of respiratory infections. MethodsA total of 60 respiratory pathogen-positive clinical samples with known redults were selected and tested using the five kits (labeled as A, B, C, D, and E). The detection rates and Kappa coefficients were calculated to evaluate the consistency between the results from these kits and those from single-pathogen PCR kits. According to the limit of detection (LOD) provided by the kits, standard substances of respiratory pathogens (including 12 types such as influenza virus, Mycoplasma pneumoniae, and Bordetella pertussis) were diluted to four concentrations (250, 500, 1 000, and 2 000 copies·mL⁻¹). All five kits were used for detection to evaluate their respective detection capabilities. ResultsCompared with the results from single-pathogen PCR kits, the five tested kits demonstrated good consistency (all Kappa >0.80). Among them, Kit A had the highest detection rate (100.00%), followed by Kits C and E (98.33%), and then Kits B and D (95.00%). All five kits showed a relatively low false negative rate (FNR) for samples with a cycle threshold (Ct) value ≤35 (≤2.38%). However, for samples with Ct values>35, the FNR increased accordingly(average FNR=6.67%, P=0.029). Kit C exhibited the highest detection sensitivity for the tested standard substances (average LOD: 458.33 copies·mL⁻¹), followed by Kit D, then Kits A/E, and finally Kit B. ConclusionThe five multiplex PCR kits showed good consistency with single-pathogen detection results, but each had its own performance emphasis. Kit A, with the highest detection rate and high throughput, is suitable for targeted viral screening. Kit B, covering the broadest pathogen spectrum (including fungi/bacteria), is suitable for comprehensive respiratory pathogen screening. Kits C, D and E, are applicable for rapid detection. It is important to note that the detection efficacy of all kits decreases for low viral load samples with Ct values >35. In practical application, selection should be based on specific screening objectives, throughput requirements, and sample types.

OBJECTIVE To formulate the Hazardous Drug List in China， and provide a scientific basis for standardizing the management of hazardous drugs， preventing and controlling occupational exposure risks in medical institutions at all levels in China. METHODS Under the joint organization of the Intravenous Medication Admixture Management Professional Committee of the Chinese Pharmaceutical Association and the National Medical Quality Control Center for Pharmaceutical Management， based on literature research and international hazardous drug lists and their development procedures， the scope of hazardous drug selection， selection principles， and evidence confirmation criteria were determined through expert panel seminars and expert questionnaire survey method （Delphi method）. Chemical drugs， biological agents， and Chinese patent medicines marketed in China that met the selecti on principles were screened to form the Chinese Expert Consensus on the Hazardous Drug List . RESULTS Five principles for selecting hazardous drugs were established （including protective labeling in package inserts， carcinogenicity， reproductive toxicity， genotoxicity， and organ toxicity at low doses）， along with the corresponding evidence confirmation criteria. The Hazardous Drug List containing 210 drugs was formulated， covering anti-tumor drugs， endocrine system drugs， nervous system drugs， immunomodulators， anti-infective drugs， cardiovascular system drugs， hematological system drugs， urinary system drugs， obstetrics and gynecology drugs， dermatological drugs and other categories. Compared with international hazardous drug lists， 25 newly included hazardous drugs were added， including 15 anti-tumor drugs， 6 Chinese patent medicines and 4 biological agents. In addition， 88 Chinese patent medicines that may have potential occupational exposure risks are listed in the appendix for reference by medical institutions in management. CONCLUSIONS The development of this consensus closely aligns with the actual clinical drug use in China， with a scientific and rigorous methodology， and can serve as a reference for medical institutions at all levels to standardize the management of hazardous drugs and prevent and control occupational exposure risks.

This study investigated the potential pathogenicity and genetic characteristics of ST314 Salmonella Kentucky(S.Ken-tucky)isolates from a broiler slaughterhouse.Antimicrobial susceptibility testing and whole-genome sequencing(WGS)were used to determine antimicrobial resistance,virulence factors,and the presence of antimicrobial resistance genes(ARGs)and mobile genetic elements(MGEs)among the isolates.The three multidrug resistant(MDR)isolates exhibited high resistance to multiple antimicrobial agents.The F4-2S strain exhibited resistance to 14 drugs across seven categories,whereas the F4T strain showed resistance to 13 drugs in the same number of categories.In contrast,the Y23 strain was resistant to nine drugs in six categories.Notably,F4-2S dem-onstrated high homology with F4T:both possessed 13 ARGs distributed across nine categories,in addition to a wide range of virulence factors,including secretion systems and effector proteins.The presence of IncR and IncX1 plasmids significantly enhanced both the antimicrobial resistance and pathogenicity of the isolates.The genome map of Y23 revealed a chromosome alongside two plasmids.The chromosome containedonly one resistance gene but several virulence factors,including the type III secretion system(T3SS),which is crucial for bacterial invasion.The plasmid pY23-1 contained eight types of 19 ARGs.Comparative analysis indicated that pY23-1 ex-hibited high homology with pZ1323SSL0055 and pSAL-045,all of which contained multiple ARGs,thus suggesting critical roles of these genes in the evolution of bacterial resistance.In conclusion,ST314 S.Kentucky demonstrated a complex mechanism of resis-tance coupled with significant pathogenic potential.The ARGs and MGEs in the plasmid contributed to the emergence and dissemina-tion of antimicrobial resistance.The multiple virulence factors present in the chromosome may be key factors driving the increasing virulence of ST314 S.Kentucky.

Purpose To investigate the correlation between intervertebral disc degeneration and paraspinal muscle fatty infiltration in chronic low back pain(CLBP)using quantitative MRI techniques.Materials and Methods A total of 63 CLBP patients and 42 volunteers in Zigong Fourth People's Hospital from September 2023 to March 2024 were retrospectively collected.Magnetic resonance scans were performed using multi-echo water-fat separation technique and T2 mapping technique.Intervertebral disc degeneration was assessed using the Pfirrmann grading,while paraspinal muscle fatty infiltration was evaluated using the Goutallier grading.Proton density fat fraction(PDFF)of the multifidus and erector spinae muscles,and T2 values of the nucleus pulposus were quantitatively measured.The relationships between these parameters and their associations with age and gender were analyzed.Results Compared with healthy volunteers,CLBP patients showed significantly reduced T2 values of the nucleus pulposusand significantly increased PDFF of the paraspinal muscles(Z=-5.54-4.65,all P<0.001).The Pfirrmann grading of intervertebral discs and the Goutallier grading of paraspinal muscles were significantly correlated with the T2 value of the nucleus pulposus and the PDFF of the paraspinal muscles(|r|>0.4,P<0.05).Age and Pfirrmann grading were independent predictors of paraspinal muscle fat infiltration(β=0.228-5.682,all P<0.001),and gender was also an independent predictor at the L4/5 intervertebral disc level(β=-2.834,-3.352,both P<0.05).Conclusion Intervertebral disc degeneration and paraspinal muscle fat infiltration are closely related in CLBP,age and gender also have an impact.

Aim To observe the interventional effects of Polygala fallax Hemsl.(PFH)on diabetic kidney disease(DKD)rats and the regulatory mechanism of ferroptosis.Methods Thirty-six SD rats were ran-domly divided into the control group,DKD group,fer-rostatin-1(Fer-1)group,PFH-L group,PFH-M group,and PFH-H group,with six rats in each group.Model-ing was induced by a one-time intraperitoneal injection of 35 mg·kg-1 streptozocin(STZ)in combination with high-sugar and high-fat feed.Ferrostatin-1(25μmol·kg-1)was injected intraperitoneally in Fer-1 group.The PFH-L,PFH-M,and PFH-H groups were gavaged with 50,100 and 200 mg·kg-1 of alcoholic extracts of PFH respectively,and the control and DKD groups were gavaged with an equal volume of distilled water once a day for eight weeks.At the end of drug administration,blood glucose,24h-UP,BUN and Scr levels were measured in each group of rats.HE stai-ning and Masson staining were used to observe renal histopathological changes.ELISA was employed to de-termine the levels of total iron,MDA and GSH activity.IHC was used to observe the expression of Nrf2,SLC7A11,and GPX4 in renal tissues.Western blot was used to detect the protein expression of COL1A1,α-SMA,TGF-,FTH-1,TFR-1,Nrf2,SLC7A1 1,GPX4,in renal tissues.RT-qPCR was used to detect the ex-pression levels of Nrf2,SLC7A11,GPX4 mRNA in re-nal tissues.Results Compared with the control group,blood glucose,24h-UP,BUN,Scr increased(P＜0.05);glomerular volume increased,tubular dilata-tion and collagen fiber deposition were obvious;total i-ron,MDA content increased and GSH activity de-creased(P＜0.05);there was increased protein ex-pression of COL1A1,α-SMA,TGF-β,TFR-1 and de-creased protein expression of FTH-1(P＜0.01);there was decreased mRNA and protein expression of Nrf2,SLC7A11,GPX4 in DKD group rats(P＜0.01).Compared with the DKD group,blood glucose,24h-UP,BUN and Scr decreased(P＜0.05);renal tissue le-sions were significantly reduced;total iron and MDA content decreased,and GSH activity increased(P＜0.05);COL1A1,α-SMA,TGF-β,and TFR-1 protein expression decreased and FTH-1 protein expression in-creased(P＜0.05,P＜0.01);Nrf2,SLC7A11,GPX4 mRNA and protein expression increased in Fer-1 and PFH dose groups(P＜0.05,P＜0.01).Conclusions PFH attenuates renal histopathological injury in DKD rats,and the mechanism may be related to the regula-tion of the Nrf2/SLC7A11/GPX4 signaling pathway.

This study investigated the potential pathogenicity and genetic characteristics of ST314 Salmonella Kentucky(S.Ken-tucky)isolates from a broiler slaughterhouse.Antimicrobial susceptibility testing and whole-genome sequencing(WGS)were used to determine antimicrobial resistance,virulence factors,and the presence of antimicrobial resistance genes(ARGs)and mobile genetic elements(MGEs)among the isolates.The three multidrug resistant(MDR)isolates exhibited high resistance to multiple antimicrobial agents.The F4-2S strain exhibited resistance to 14 drugs across seven categories,whereas the F4T strain showed resistance to 13 drugs in the same number of categories.In contrast,the Y23 strain was resistant to nine drugs in six categories.Notably,F4-2S dem-onstrated high homology with F4T:both possessed 13 ARGs distributed across nine categories,in addition to a wide range of virulence factors,including secretion systems and effector proteins.The presence of IncR and IncX1 plasmids significantly enhanced both the antimicrobial resistance and pathogenicity of the isolates.The genome map of Y23 revealed a chromosome alongside two plasmids.The chromosome containedonly one resistance gene but several virulence factors,including the type III secretion system(T3SS),which is crucial for bacterial invasion.The plasmid pY23-1 contained eight types of 19 ARGs.Comparative analysis indicated that pY23-1 ex-hibited high homology with pZ1323SSL0055 and pSAL-045,all of which contained multiple ARGs,thus suggesting critical roles of these genes in the evolution of bacterial resistance.In conclusion,ST314 S.Kentucky demonstrated a complex mechanism of resis-tance coupled with significant pathogenic potential.The ARGs and MGEs in the plasmid contributed to the emergence and dissemina-tion of antimicrobial resistance.The multiple virulence factors present in the chromosome may be key factors driving the increasing virulence of ST314 S.Kentucky.

Aim To observe the interventional effects of Polygala fallax Hemsl.(PFH)on diabetic kidney disease(DKD)rats and the regulatory mechanism of ferroptosis.Methods Thirty-six SD rats were ran-domly divided into the control group,DKD group,fer-rostatin-1(Fer-1)group,PFH-L group,PFH-M group,and PFH-H group,with six rats in each group.Model-ing was induced by a one-time intraperitoneal injection of 35 mg·kg-1 streptozocin(STZ)in combination with high-sugar and high-fat feed.Ferrostatin-1(25μmol·kg-1)was injected intraperitoneally in Fer-1 group.The PFH-L,PFH-M,and PFH-H groups were gavaged with 50,100 and 200 mg·kg-1 of alcoholic extracts of PFH respectively,and the control and DKD groups were gavaged with an equal volume of distilled water once a day for eight weeks.At the end of drug administration,blood glucose,24h-UP,BUN and Scr levels were measured in each group of rats.HE stai-ning and Masson staining were used to observe renal histopathological changes.ELISA was employed to de-termine the levels of total iron,MDA and GSH activity.IHC was used to observe the expression of Nrf2,SLC7A11,and GPX4 in renal tissues.Western blot was used to detect the protein expression of COL1A1,α-SMA,TGF-,FTH-1,TFR-1,Nrf2,SLC7A1 1,GPX4,in renal tissues.RT-qPCR was used to detect the ex-pression levels of Nrf2,SLC7A11,GPX4 mRNA in re-nal tissues.Results Compared with the control group,blood glucose,24h-UP,BUN,Scr increased(P＜0.05);glomerular volume increased,tubular dilata-tion and collagen fiber deposition were obvious;total i-ron,MDA content increased and GSH activity de-creased(P＜0.05);there was increased protein ex-pression of COL1A1,α-SMA,TGF-β,TFR-1 and de-creased protein expression of FTH-1(P＜0.01);there was decreased mRNA and protein expression of Nrf2,SLC7A11,GPX4 in DKD group rats(P＜0.01).Compared with the DKD group,blood glucose,24h-UP,BUN and Scr decreased(P＜0.05);renal tissue le-sions were significantly reduced;total iron and MDA content decreased,and GSH activity increased(P＜0.05);COL1A1,α-SMA,TGF-β,and TFR-1 protein expression decreased and FTH-1 protein expression in-creased(P＜0.05,P＜0.01);Nrf2,SLC7A11,GPX4 mRNA and protein expression increased in Fer-1 and PFH dose groups(P＜0.05,P＜0.01).Conclusions PFH attenuates renal histopathological injury in DKD rats,and the mechanism may be related to the regula-tion of the Nrf2/SLC7A11/GPX4 signaling pathway.

Aim To investigate the effects of Mdivi-1 on A1 astrocyte activation and its associated signaling molecules.Methods CTX-TNA2 astrocytes were di-vided into the control,ACM,and low-,medium-,and high-dose Mdivi-1 groups based on concentration screening via CCK-8 assay.ACM,a DMEM high-glu-cose medium containing preset concentrations of IL-1α,TNF-α,and C1q,was used to induce A1 activa-tion.The ACM group was stimulated with ACM for 24 hours.Mdivi-1 groups were pretreated with correspond-ing concentrations of Mdivi-1 for 2 hours,followed by ACM stimulation for 24 hours.Real-time quantitative PCR and Western blot were employed to assess mRNA levels and protein expression of IL-1β,C3,and iNOS in all groups.Immunofluorescence and Western blot were used to detect the expression of signaling molecules NLRP3,caspase-1,and ASC.DHE labeling was used to assess and flow cytometry was used to examine reac-tive oxygen species(ROS)levels.Results The CCK-8 assay identified 5,10,and 25 μmol·L-1as ap-propriate concentrations for Mdivi-1 intervention in CTX-TNA2 cells.Real-time quantitative PCR and Western blot results indicated that,compared to the control group,IL-1 β,C3,and iNOS mRNA levels and protein expression were significantly elevated in the ACM group(P＜0.05).In contrast,these levels were significantly reduced in the 10 and 25 μmnol·L-1 Mdi-vi-1 groups compared to the ACM group(P＜0.05).Immunofluorescence and Western blot results confirmed that ACM stimulation significantly activated the NLRP3 inflammasome in astrocytes,while Mdivi-1 intervention effectively reversed the ACM-induced upregulation of NLRP3,caspase-1,and ASC.DHE staining results demonstrated that 5,10,and 25 μmol·L-1Mdivi-1 in-terventions partially reversed the ACM-induced in-crease in ROS levels in a dose-dependent manner.Conclusion Mdivi-1 effectively inhibits A1 astrocyte activation,potentially through modulation of ROS and the NLRP3 inflammasomes.

Aim To compare the observation results of atomic force microscopy(AFM)and scanning electron microscopy(SEM),and to summarize the main problems and solutions of AFM in observing biological macromolecules,using the observa-tion subjects of protein samples purified by our research group.Methods The protein samples were diluted to 15 nmol·L-1 with PBS,fixed on glass slides,silicon wafers,and mica sheets,dried,and made into solid-phase observation samples.SEM sam-ples were plated with platinum before observation.The surface structures of proteins were observed using AFM and SEM,sample heights were calculated,and differences in results were com-pared.Results Protein samples with positive charges tended to shift to the right during observation due to the repulsion of the AFM probe;mica sheets could effectively eliminate the positive charge of proteins to avoid sample movement;PBS provided a stable environment for protein samples,but the crystallization of PBS salts interfered with probe operation and imaging clarity;SEM samples needed to be plated with platinum before observa-tion and could not achieve the precision of AFM.Conclusions Both AFM and SEM can directly observe protein structures in vitro,with AFM providing higher precision results;when protein sample stability permits,ultrapure water is preferred as the sol-vent carrier,and volatile liquids such as ethanol can also serve as solvent carriers.The application of AFM offers a new approach for pharmacological studies on interactions between biological macromolecules.