This study aims to explore the effects and action mechanisms of the active ingredients in Buyang Huanwu Decoction(BYHWD), namely tetramethylpyrazine(TMP) and hydroxy-safflor yellow A(HSYA), on oxygen-glucose deprivation/reglucose-reoxygenation(OGD/R)-induced inflammation and oxidative stress of microglia(MG). Network pharmacology was used to screen the effective monomer ingredients of BYHWD and determine the safe concentration range for each component. Inflammation and oxidative stress models were established to further screen the best ingredient combination and optimal concentration ratio with the most effective anti-inflammatory and antioxidant effects. OGD/R BV2 cell models were constructed, and BV2 cells in the logarithmic growth phase were divided into a normal group, a model group, an HSYA group, a TMP group, and an HSYA + TMP group. Enzyme-linked immunosorbent assay(ELISA) was used to detect the levels of inflammatory cytokines such as interleukin-1β(IL-1β), tumor necrosis factor-α(TNF-α), and interleukin-6(IL-6). Oxidative stress markers, including superoxide dismutase(SOD), nitric oxide(NO), and malondialdehyde(MDA), were also measured. Western blot was used to analyze the protein expression of both inflammation-related pathway [Toll-like receptor 4(TLR4)/nuclear factor-kappa B(NF-κB)] and oxidative stress-related pathway [nuclear factor erythroid 2-related factor 2(Nrf2)/heme oxygenase-1(HO-1)]. Immunofluorescence was used to assess the expression of proteins such as inducible nitric oxide synthase(iNOS) and arginase-1(Arg-1). The most effective ingredients for anti-inflammatory and antioxidant effects in BYHWD were TMP and HSYA. Compared to the normal group, the model group showed significantly increased levels of IL-1β, TNF-α, IL-6, NO, and MDA, along with significantly higher protein expression of NF-κB, TLR4, Nrf2, and HO-1 and significantly lower SOD levels. The differences between the two groups were statistically significant. Compared to the model group, both the HSYA group and the TMP group showed significantly reduced levels of IL-1β, TNF-α, IL-6, NO, and MDA, lower expression of NF-κB and TLR4 proteins, higher levels of SOD, and significantly increased protein expression of Nrf2 and HO-1. Additionally, the expression of the M1-type MG marker iNOS was significantly reduced, while the expression of the M2-type MG marker Arg-1 was significantly increased. The results of the HSYA group and the TMP group had statistically significant differences from those of the model group. Compared to the HSYA group and the TMP group, the HSYA + TMP group showed further significant reductions in IL-1β, TNF-α, IL-6, NO, and MDA levels, along with significant reductions in NF-κB and TLR4 protein expression, an increase in SOD levels, and elevated Nrf2 and HO-1 protein expression. Additionally, the expression of the M1-type MG marker iNOS was reduced, while the M2-type MG marker Arg-1 expression increased significantly in the HSYA + TMP group compared to the TMP or HSYA group. The differences in the results were statistically significant between the HSYA + TMP group and the TMP or HSYA group. The findings indicated that the combined use of HSYA and TMP, the active ingredients of BYHWD, can effectively inhibit OGD/R-induced inflammation and oxidative stress of MG, showing superior effects compared to the individual use of either component.
Oxidative Stress/drug effects* ; Drugs, Chinese Herbal/pharmacology* ; Animals ; Mice ; Glucose/metabolism* ; Cell Line ; Inflammation/genetics* ; Oxygen/metabolism* ; Pyrazines/pharmacology* ; Microglia/metabolism* ; NF-E2-Related Factor 2/immunology* ; NF-kappa B/immunology* ; Toll-Like Receptor 4/immunology* ; Anti-Inflammatory Agents/pharmacology* ; Humans

Objective:To explore the clinical features of Brucellar myelitis and diagnosis and treatment of secondary neuromyelitis optica spectrum disorder (NMOSD), and enhance the awareness of clinicians about this disease.Methods:A retrospective study was performed; 13 patients with Brucellar myelitis admitted to Department of Neurology, Inner Mongolia Autonomous Region People's Hospital from January 2020 to December 2024 were chosen. Clinical data were collected, and MRI images and serological changes during the infection period were observed. Serum and cerebrospinal fluid demyelinating antibody markers and cerebrospinal fluid oligoclonal bands (OCBs) in the suspected secondary inflammatory demyelinating diseases of the central nervous system patients were detected. All patients received standard antibiotic treatment and/or individualized immunotherapy depending on disease severity. The patients were followed up for 24 (12, 42) months. At the last follow-up, the neurological outcomes were evaluated using modified Rankin scale (mRS, scores of 0-2: good prognosis; scores of 3-6: poor prognosis).Results:(1) Among the 13 patients, 12 had motor disorder, 9 had bladder/bowel dysfunction, 7 had sensory abnormality, and 4 had other symptoms such as dizziness, behavioral changes, or unsteady gait. (2) MRI results showed that 8 patients had spinal cord abnormalities, including 2 with long-segment intramedullary high signal at T2-weighted image and 6 with short-segment local intramedullary high signal at T2-weighted image. Enhanced MRI was performed in 11 patients, with 2 showing lesion enhancement, 3 showing meningeal enhancement, and 6 showing no enhancement. (3) Four patients had elevated cerebrospinal fluid pressure (>180 mmH 2O); 9 patients had elevated cerebrospinal fluid protein level (>0.45 g/L). Brucella-specific DNA was detected in the cerebrospinal fluid of 6 patients. One patient was positive for OCB type II. One patient was positive for aquaporin 4 antibody (AQP4-IgG) in both serum and cerebrospinal fluid, and one patient was double positive for myelin oligodendrocyte glycoprotein antibody (MOG-IgG) and AQP4-IgG in serum. (4) All 13 patients received standard antibiotic treatment; 12 patients received immunotherapy. (5) Among the 4 patients with poor prognosis, 3 died and the remaining 9 had a good prognosis. The mRS score decreasing from 4 (3, 4) at admission to 2 (2, 3) at the last follow-up, showing an overall improvement in neurological function. (6) Among the 13 patients, 2 were diagnosed as having Brucellar myelitis secondary NMOSD. On the basis of antibiotic treatment, one AQP4-IgG positive patient was treated with high-dose glucocorticoids only and later died; one MOG-IgG and AQP4-IgG double positive patient was treated with intravenous immunoglobulin combined with high-dose glucocorticoids and sequential rituximab, with mRS score decreasing from 5 at admission to 2 at the last follow-up and good neurological function recovery. Conclusions:The clinical manifestations of Brucellar myelitis are diverse and overlap with the clinical features of NMOSD. For patients with suspected Brucellar myelitis secondary NMOSD, combination of immunosuppressant (such as rituximab) with antibiotics may be an effective individualized treatment.

Objective To analyze the distribution and drug resistance patterns of pathogenic bacteria in bile and blood cultures obtained from patients with biliary stones accompanied by acute biliary tract infection,to evaluate the clinical appropriate-ness of antibiotic use based on drug sensitivity results,and to provide evidence for empirical antibiotic treatment in such patients.Methods The clinical data of 161 patients with biliary calculi complicated by acute biliary tract infection who were admitted to the First People's Hospital of Neijiang from 2017 to 2023 were retrospectively analyzed.The results of microbial culture,drug sensitivity analysis,and patient characteristics were assessed to evaluate the appropriateness of clinical antimicrobial therapy.Results Among the 161 patients with positive cultures,212 strains of pathogenic bacteria were detected.The predominant patho-gens were Escherichia coli,Klebsiella pneumoniae subspecies,and Enterococcus faecium.Age and underlying diseases significantly affected the distribution of Escherichia coli and Klebsiella pneumoniae subspecies.Within the gram-negative bacterial group,Esche-richia coli and Klebsiella pneumoniae subspecies exhibited higher drug resistance to commonly used broad-spectrum penicillin,third-generation cephalosporin and quinolones but lower resistance rates to piperacillin and tazobactam;furthermore,elderly indi-viduals aged ≥65 years showed higher resistance rates to ceftriaxone than those under age 65 while people with drug exposure history had higher ceftazidime resistance rates that were statistically significant.In contrast to Enterococcus faecalis which displayed low antimicrobial resistance rates for most drugs tested in this study,Enterococcus faecium demonstrated high levels of antibiotic resistance;however,both Enterococcus faecalis and Enterococcus faecium exhibited zero-resistance rates against vancomycin and tigecycline although this may be attributed to their small sample size in our study cohort.Finally,we found that empirical anti-in-fective drugs,as well as target anti-infective drugs,were not prescribed rationally among these patients due mainly to inappropriate combinations of antibiotics or incorrect dosages.Conclusions The predominant pathogens in patients with acute biliary tract infection are gram-negative bacteria,Gram-positive bacteria,and fungi;however,the potential involvement of anaerobic bacteria should not be overlooked.Vancomycin exhibits sensitivity against gram-positive bacteria,yet the overall rationality of antibiotic usage remains suboptimal.Enhanced clinical testing for pathogenic microorganisms is imperative in the management of biliary stones accompanied by acute biliary tract infection.In contrast,clinical pharmacists should provide comprehensive training on anti-infective drugs to clinicians to facilitate their judicious selection of antibiotics based on drug sensitivity results and prevent the e-mergence of multidrug-resistant bacteria.

Objective:To investigate the role of resveratrol(RES)in improving airway inflammation in children with asthma by regulating miR-125b-5p/PRDM1 axis and its molecular mechanism.Methods:Rat model of asthma was induced by ovalbumin(OVA).The rats were randomly divided into sham group(sham),asthma group(model),10 μmol/L RES(RES-L)group,50 μmol/L RES(RES-H)group,50 μmol/L miR-125b-5p antagomir(miR-125b-5p antagomir)group and miR-125b-5p antagomir+50 μmol/L RES(miR-125b-5p antagomir+RES)group.The number of inflammatory cells in bronchoalveolar lavage fluid(BALF)was detected.HE staining and Masson three-color staining were used to observe pathological changes of lung tissue.Levels of inflammatory factors in lung homogenates were detected by ELISA.Serum from asthmatic children and healthy children was collected for analysis.Expressions of miR-125b-5p and PRDM1 in lung tissues of asthmatic and healthy children and different groups of rats were detected by RT-qPCR.Targeted binding relationship between miR-125b-5p and PRDM1 was detected by double luciferase assay.Results:HE staining showed that the airway epithelium in asthma group had a variety of inflammatory cell infiltration,while pathological changes in RES group were reduced.Masson staining showed that the area of airway collagen deposition was reduced in RES group compared with asth-ma group.The number of inflammatory cells in BALF in asthma group was higher than that in RES group.RES treatment significantly reduced airway wall and smooth muscle thickness.ELISA results showed that lung tissue in the asthma group had increased levels of IL-1β,IL-10 and TNF-α,while decreased level of IL-12,the RES group showed the opposite trend.Level of miR-125b-5p in lung tissue of asthmatic rats was significantly increased,while level of PRDM1 was decreased,and RES treatment could reverse this result.In addi-tion,treatment with miR-125b-5p antagomir alleviated airway inflammation and airway remodeling in asthmic rats.Dual luciferase assay confirmed the targeting binding relationship between miR-125b-5p and PRDM1.Compared with healthy children,serum levels of IL-1β,IL-10 and TNF-α were increased,while IL-12 level was decreased in children with asthma.Conclusion:RES inhibits the re-lease of inflammatory cytokines through HMGB1/TLR4/NF-κB pathway,alleviating asthma-induced airway inflammation and airway remodeling.

Objective To analyze the distribution and drug resistance patterns of pathogenic bacteria in bile and blood cultures obtained from patients with biliary stones accompanied by acute biliary tract infection,to evaluate the clinical appropriate-ness of antibiotic use based on drug sensitivity results,and to provide evidence for empirical antibiotic treatment in such patients.Methods The clinical data of 161 patients with biliary calculi complicated by acute biliary tract infection who were admitted to the First People's Hospital of Neijiang from 2017 to 2023 were retrospectively analyzed.The results of microbial culture,drug sensitivity analysis,and patient characteristics were assessed to evaluate the appropriateness of clinical antimicrobial therapy.Results Among the 161 patients with positive cultures,212 strains of pathogenic bacteria were detected.The predominant patho-gens were Escherichia coli,Klebsiella pneumoniae subspecies,and Enterococcus faecium.Age and underlying diseases significantly affected the distribution of Escherichia coli and Klebsiella pneumoniae subspecies.Within the gram-negative bacterial group,Esche-richia coli and Klebsiella pneumoniae subspecies exhibited higher drug resistance to commonly used broad-spectrum penicillin,third-generation cephalosporin and quinolones but lower resistance rates to piperacillin and tazobactam;furthermore,elderly indi-viduals aged ≥65 years showed higher resistance rates to ceftriaxone than those under age 65 while people with drug exposure history had higher ceftazidime resistance rates that were statistically significant.In contrast to Enterococcus faecalis which displayed low antimicrobial resistance rates for most drugs tested in this study,Enterococcus faecium demonstrated high levels of antibiotic resistance;however,both Enterococcus faecalis and Enterococcus faecium exhibited zero-resistance rates against vancomycin and tigecycline although this may be attributed to their small sample size in our study cohort.Finally,we found that empirical anti-in-fective drugs,as well as target anti-infective drugs,were not prescribed rationally among these patients due mainly to inappropriate combinations of antibiotics or incorrect dosages.Conclusions The predominant pathogens in patients with acute biliary tract infection are gram-negative bacteria,Gram-positive bacteria,and fungi;however,the potential involvement of anaerobic bacteria should not be overlooked.Vancomycin exhibits sensitivity against gram-positive bacteria,yet the overall rationality of antibiotic usage remains suboptimal.Enhanced clinical testing for pathogenic microorganisms is imperative in the management of biliary stones accompanied by acute biliary tract infection.In contrast,clinical pharmacists should provide comprehensive training on anti-infective drugs to clinicians to facilitate their judicious selection of antibiotics based on drug sensitivity results and prevent the e-mergence of multidrug-resistant bacteria.

Objective:To investigate the role of resveratrol(RES)in improving airway inflammation in children with asthma by regulating miR-125b-5p/PRDM1 axis and its molecular mechanism.Methods:Rat model of asthma was induced by ovalbumin(OVA).The rats were randomly divided into sham group(sham),asthma group(model),10 μmol/L RES(RES-L)group,50 μmol/L RES(RES-H)group,50 μmol/L miR-125b-5p antagomir(miR-125b-5p antagomir)group and miR-125b-5p antagomir+50 μmol/L RES(miR-125b-5p antagomir+RES)group.The number of inflammatory cells in bronchoalveolar lavage fluid(BALF)was detected.HE staining and Masson three-color staining were used to observe pathological changes of lung tissue.Levels of inflammatory factors in lung homogenates were detected by ELISA.Serum from asthmatic children and healthy children was collected for analysis.Expressions of miR-125b-5p and PRDM1 in lung tissues of asthmatic and healthy children and different groups of rats were detected by RT-qPCR.Targeted binding relationship between miR-125b-5p and PRDM1 was detected by double luciferase assay.Results:HE staining showed that the airway epithelium in asthma group had a variety of inflammatory cell infiltration,while pathological changes in RES group were reduced.Masson staining showed that the area of airway collagen deposition was reduced in RES group compared with asth-ma group.The number of inflammatory cells in BALF in asthma group was higher than that in RES group.RES treatment significantly reduced airway wall and smooth muscle thickness.ELISA results showed that lung tissue in the asthma group had increased levels of IL-1β,IL-10 and TNF-α,while decreased level of IL-12,the RES group showed the opposite trend.Level of miR-125b-5p in lung tissue of asthmatic rats was significantly increased,while level of PRDM1 was decreased,and RES treatment could reverse this result.In addi-tion,treatment with miR-125b-5p antagomir alleviated airway inflammation and airway remodeling in asthmic rats.Dual luciferase assay confirmed the targeting binding relationship between miR-125b-5p and PRDM1.Compared with healthy children,serum levels of IL-1β,IL-10 and TNF-α were increased,while IL-12 level was decreased in children with asthma.Conclusion:RES inhibits the re-lease of inflammatory cytokines through HMGB1/TLR4/NF-κB pathway,alleviating asthma-induced airway inflammation and airway remodeling.

Objective:To explore the clinical features of Brucellar myelitis and diagnosis and treatment of secondary neuromyelitis optica spectrum disorder (NMOSD), and enhance the awareness of clinicians about this disease.Methods:A retrospective study was performed; 13 patients with Brucellar myelitis admitted to Department of Neurology, Inner Mongolia Autonomous Region People's Hospital from January 2020 to December 2024 were chosen. Clinical data were collected, and MRI images and serological changes during the infection period were observed. Serum and cerebrospinal fluid demyelinating antibody markers and cerebrospinal fluid oligoclonal bands (OCBs) in the suspected secondary inflammatory demyelinating diseases of the central nervous system patients were detected. All patients received standard antibiotic treatment and/or individualized immunotherapy depending on disease severity. The patients were followed up for 24 (12, 42) months. At the last follow-up, the neurological outcomes were evaluated using modified Rankin scale (mRS, scores of 0-2: good prognosis; scores of 3-6: poor prognosis).Results:(1) Among the 13 patients, 12 had motor disorder, 9 had bladder/bowel dysfunction, 7 had sensory abnormality, and 4 had other symptoms such as dizziness, behavioral changes, or unsteady gait. (2) MRI results showed that 8 patients had spinal cord abnormalities, including 2 with long-segment intramedullary high signal at T2-weighted image and 6 with short-segment local intramedullary high signal at T2-weighted image. Enhanced MRI was performed in 11 patients, with 2 showing lesion enhancement, 3 showing meningeal enhancement, and 6 showing no enhancement. (3) Four patients had elevated cerebrospinal fluid pressure (>180 mmH 2O); 9 patients had elevated cerebrospinal fluid protein level (>0.45 g/L). Brucella-specific DNA was detected in the cerebrospinal fluid of 6 patients. One patient was positive for OCB type II. One patient was positive for aquaporin 4 antibody (AQP4-IgG) in both serum and cerebrospinal fluid, and one patient was double positive for myelin oligodendrocyte glycoprotein antibody (MOG-IgG) and AQP4-IgG in serum. (4) All 13 patients received standard antibiotic treatment; 12 patients received immunotherapy. (5) Among the 4 patients with poor prognosis, 3 died and the remaining 9 had a good prognosis. The mRS score decreasing from 4 (3, 4) at admission to 2 (2, 3) at the last follow-up, showing an overall improvement in neurological function. (6) Among the 13 patients, 2 were diagnosed as having Brucellar myelitis secondary NMOSD. On the basis of antibiotic treatment, one AQP4-IgG positive patient was treated with high-dose glucocorticoids only and later died; one MOG-IgG and AQP4-IgG double positive patient was treated with intravenous immunoglobulin combined with high-dose glucocorticoids and sequential rituximab, with mRS score decreasing from 5 at admission to 2 at the last follow-up and good neurological function recovery. Conclusions:The clinical manifestations of Brucellar myelitis are diverse and overlap with the clinical features of NMOSD. For patients with suspected Brucellar myelitis secondary NMOSD, combination of immunosuppressant (such as rituximab) with antibiotics may be an effective individualized treatment.

Objective:To evaluate the safety and efficacy of 225Ac-prostate specific membrane antigen (PSMA) in the treatment of metastatic castration-resistant prostate cancer (mCRPC). Methods:Eleven patients (age (70.0±8.8) years) with mCRPC who were treated with 225Ac-PSMA-617 between July 2021 and October 2023 in the Affiliated Hospital of Southwest Medical University were retrospectively analyzed. In order to assess efficacy, the Prostate Cancer Clinical Trials Working Group 3 (PCWG3) criteria were used to evaluate the changes in prostate specific antigen (PSA) level after the treatment. 68Ga-PSMA-11 PET/CT imaging was performed at the baseline and after the treatment, and molecular imaging response was assessed using the modified PET response criteria in solid tumors (PERCIST) 1.0. Progression-free survival (PFS) and overall survival (OS) were analyzed using the Kaplan-Meier method. Toxicity was assessed by common terminology criteria for adverse events version 5.0 (CTCAE 5.0). The paired t test or Wilcoxon signed rank test was used to compare the parameters before and after treatment. Results:Post-treatment PSA levels were significantly lower than pre-treatment in 9 of 11 patients (17.83(4.74, 41.25) vs 124.33(77.85, 784.22) μg/L; z=-2.67, P=0.008), and 6 of them decreasing by more than 50% and 2 patients experienced progressive disease (PD) with PSA levels rising by more than 25%. Post-treatment 68Ga-PSMA-11 PET/CT showed that 7 patients achieved partial response (PR), 2 patients achieved stable disease (SD), and 2 patients were with PD. The OS was 12.0(10.0, 18.0) months and PFS was 8.0(6.0, 11.0) months in 11 patients. There were no statistically significant differences after therapy in WBC counts, Hb, PLT, creatinine, glomerular filtration rate, alanine aminotransferase, aspartate aminotransferase, total bilirubin ( z values: from -1.07 to 0.00, t values: from -0.77 to 1.76, all P>0.05). No grade Ⅲ/Ⅳ nephrotoxicity or salivary gland toxicity was observed. Conclusion:225Ac-PSMA-617 is a promising novel therapeutic option for mCRPC with favorable safety and tolerability.

Objective To investigate changes in the urinary metabolite profiles of children exposed to polycyclic aromatic hydrocarbons(PAHs)during critical brain development and explore their potential link with the intestinal microbiota. Methods Liquid chromatography-tandem mass spectrometry was used to determine ten hydroxyl metabolites of PAHs(OH-PAHs)in 36-month-old children.Subsequently,37 children were categorized into low-and high-exposure groups based on the sum of the ten OH-PAHs.Ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry was used to identify non-targeted metabolites in the urine samples.Furthermore,fecal flora abundance was assessed by 16S rRNA gene sequencing using Illumina MiSeq. Results The concentrations of 21 metabolites were significantly higher in the high exposure group than in the low exposure group(variable importance for projection>1,P<0.05).Most of these metabolites were positively correlated with the hydroxyl metabolites of naphthalene,fluorine,and phenanthrene(r=0.336-0.531).The identified differential metabolites primarily belonged to pathways associated with inflammation or proinflammatory states,including amino acid,lipid,and nucleotide metabolism.Additionally,these distinct metabolites were significantly associated with specific intestinal flora abundances(r=0.34-0.55),which were mainly involved in neurodevelopment. Conclusion Higher PAH exposure in young children affected metabolic homeostasis,particularly that of certain gut microbiota-derived metabolites.Further investigation is needed to explore the potential influence of PAHs on the gut microbiota and their possible association with neurodevelopmental outcomes.

Objective:To investigate the effects of selinexor,a inhibitor of nuclear export protein 1(XPO1)on the proliferation inhibition and apoptosis of Kasumi-1 cells in acute myeloid leukemia(AML).Methods:MTS method was used to detect the inhibitory effect of different concentrations of selinexor on the proliferation of Kasumi-1 cells at different time points.The apoptosis rate and cell cycle changes after treatment with different concentration of selinexor were detected by flow cytometry.Results:Selinexor inhibited the growth of Kasumi-1 cells at different time points in a concentration-dependent manner(r24 h=0.7592,r48 h=0.9456,and r72 h=0.9425).Selinexor inhibited Kasumi-1 cells growth in a time-dependent manner(r=0.9057 in 2.5 μmol/L group,r=0.9897 in 5 μmol/L group and r=0.9994 in 10 μmol/L group).Selinexor could induce apoptosis of Kasumi-1 cells in a dose-dependent manner(r=0.9732),and the apoptosis of Kasumi-1 cells was more obvious with the increase of drug concentration.The proportion of G0/G1 phase was significantly increased and the proportion of S phase was significantly decreased after the treatment of Kasumi-1 cells by selinexor.With the increase of drug concentration,the proportion of Kasumi-1 cells cycle arrest in G0/G1 phase was increased and the cell synthesis was decreased.Conclusion:Selinexor can promote the death of tumor cells by inhibiting Kasumi-1 cells proliferation,inducing apoptosis and blocking cell cycle.