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

The efficacy mechanism of the alcoholic extract of Gnaphalium affine was investigated by observing its influence on oxidative stress and intestinal flora in rats modeled for chronic obstructive pulmonary disease(COPD). UPLC-MS was used to evaluate the quality of the alcoholic extract of G. affine, and 72 rats were randomly divided into six groups, with COPD models established in five groups by cigarette smoke combined with airway drip lipopolysaccharide, and the rats were given the positive drug of Danlong Oral Solution, as well as low-, medium-, and high-doses alcoholic extract of G. affine, respectively. After two weeks of continuous gastric gavage, the body weights and general morphology observations were performed; HE staining and Masson staining were used to verify the effects of the alcoholic extract of G. affine on alveolar inflammation and collagen deposition area in COPD rats; the oxidative stress indexes CAT and GSH in serum and SOD and MDA in lung tissue of the rats were measured, and the mRNA expression of HO-1, Nrf2, and NQO1 were determined by qRT-PCR. The protein expressions of HO-1, Nrf2, and NQO1 were determined by the Western blot method, and the mechanism by which the alcoholic extract of G. affine affected oxidative stress in COPD rats was explored. Finally, the influence of G. affine on the changes in intestinal flora caused by COPD was studied by 16S rRNA sequencing. The results showed that a total of 121 chemical components were identified by UPLC-MS, including 70 positive and 51 negative ion modes. In animal experiments, it was found that the alcoholic extracts of G. affine were able to reduce the percentage of collagen deposition, affect the oxidative stress indexes such as CAT, GSH, SOD, MDA, as well as the mRNA and protein expression of Nrf2, HO-1, and NQO1. The 16S rRNA sequencing results showed an increase in the level of Lactobacillales and a decrease in the level of Desulfovibrio and Desulfovibrionales, suggesting that the alcoholic extracts of G. affine could reverse the changes in intestinal flora caused by COPD. In conclusion, the alcoholic extracts of G. affine may exert anti-COPD effects by affecting the oxidative stress pathway and modulating the changes in intestinal flora.
Animals ; Oxidative Stress/drug effects* ; Pulmonary Disease, Chronic Obstructive/genetics* ; Rats ; Male ; Gastrointestinal Microbiome/drug effects* ; Rats, Sprague-Dawley ; Drugs, Chinese Herbal/administration & dosage* ; NF-E2-Related Factor 2/metabolism* ; Humans ; Lung/metabolism*

OBJECTIVES: To report the development, validation, and findings of the Multi-dimensional Attention Rating Scale (MARS), a self-report tool crafted to evaluate six-dimension attention levels. METHODS: The MARS was developed based on Classical Test Theory (CTT). Totally 202 highly educated healthy adult participants were recruited for reliability and validity tests. Reliability was measured using Cronbach's alpha and test-retest reliability. Structural validity was explored using principal component analysis. Criterion validity was analyzed by correlating MARS scores with the Toronto Hospital Alertness Test (THAT), the Attentional Control Scale (ACS), and the Attention Network Test (ANT). RESULTS: The MARS comprises 12 items spanning six distinct dimensions of attention: focused attention, sustained attention, shifting attention, selective attention, divided attention, and response inhibition.As assessed by six experts, the content validation index (CVI) was 0.95, the Cronbach's alpha for the MARS was 0.78, and the test-retest reliability was 0.81. Four factors were identified (cumulative variance contribution rate 68.79%). The total score of MARS was correlated positively with THAT (r = 0.60, P < 0.01) and ACS (r = 0.78, P < 0.01) and negatively with ANT's reaction time for alerting (r = -0.31, P = 0.049). CONCLUSIONS The MARS can reliably and validly assess six-dimension attention levels in real-world settings and is expected to be a new tool for assessing multi-dimensional attention impairments in different mental disorders.
Humans ; Adult ; Male ; Attention/physiology* ; Female ; Middle Aged ; Reproducibility of Results ; Young Adult ; Psychometrics

BACKGROUND:Bone morphogenetic protein 2 is vital in embryonic development,bone formation,and regeneration,but its high-dose application is linked to cancer.Bone morphogenetic protein 2 osteogenic peptide L20 reduces adverse effects like cancer and boosts bone tissue regeneration.OBJECTIVE:To graft bone morphogenetic protein 2 active peptide segments onto mesopores and surfaces through a peptide mimicry strategy inspired by oysters,and explore its impact on osteogenic properties of tissue-engineered bone.METHODS:(1)Mesoporous bioactive glass was synthesized using a template method.Bone morphogenetic protein 2 osteogenic peptide L20 was loaded onto mesoporous bioactive glass using a one-step synthesis method to characterize the morphology and in vitro sustained release properties of mesoporous active glass nanoparticles loaded with bone morphogenetic protein 2 osteogenic active peptide L20.(2)Bone marrow mesenchymal stem cells were isolated and extracted from SD rats.After two generations,they were co-cultured with PBS(blank group),mesoporous bioactive glass nanoparticles(control group),and mesoporous bioactive glass nanoparticles loaded with bone morphogenetic protein 2 osteogenic active peptide L20(experimental group).Cell live/dead fluorescence staining and CCK-8 assay were used to detect cytotoxicity and cell proliferation.Scanning electron microscopy was used to observe cell adhesion.After osteogenic induction and differentiation,alkaline phosphatase staining,Alizarin red S staining,and osteogenesis-related gene expression were detected.(3)Fifteen SD rats were selected to establish bilateral femoral condyle defect models and divided into three groups using a random number table method:the blank group(n=5)was not implanted with any material;the control group(n=5)was implanted with mesoporous bioactive glass nanoparticles,and the experimental group(n=5)was implanted with mesoporous bioactive glass nanoparticles loaded with bone morphogenetic protein 2 osteogenic active peptide L20.Eight weeks after surgery,femoral Micro-CT scanning and tissue morphology observation were performed.RESULTS AND CONCLUSION:(1)Scanning electron microscopy showed that the mesoporous bioactive glass nanoparticles loaded with bone morphogenetic protein 2 osteogenic active peptide L20 were spherical and monodisperse particles.Transmission electron microscopy showed their porous structure with an average particle size of(268.10±0.58)nm,which could release L20 in vitro.(2)Mesoporous bioglass nanoparticles loaded with bone morphogenetic protein 2 osteogenic active peptide L20 were non-cytotoxic and could promote the proliferation and adhesion of bone marrow mesenchymal stem cells.Compared with the blank group and the control group,the alkaline phosphatase activity and extracellular matrix mineralization capacity of the experimental group were increased(P<0.05),and the mRNA expression levels of alkaline phosphatase,Runx2,and osteocalcin were increased(P<0.05).(3)The results of femoral Micro-CT scanning showed that compared with the blank group and the control group,the new bone mass and bone density of the experimental group were increased(P<0.05).The results of hematoxylin-eosin and Masson staining showed that compared with the blank group and the control group,the new bone formation and collagen fibers of the experimental group were increased.(4)These findings indicate that mesoporous bioactive glass loaded with bone morphogenetic protein 2 active peptide L20 exhibits excellent biocompatibility and in vitro and in vivo osteogenic properties,promoting regeneration and repair of SD rat femoral condyle defects.

Objective To explore the safety and effects of alpha-lipoic acid (ALA) in patients with ischemic heart failure (IHF). Methods A randomized, double-blind, placebo-controlled trial was designed (ClinicalTrial.gov registration number NCT03491969). From January 2019 to January 2023, 300 patients with IHF were enrolled in four medical centers in China, and were randomly assigned at a 1∶1 ratio to receive ALA (600 mg daily) or placebo on top of standard care for 24 months. The primary outcome was the composite outcome of hospitalization for heart failure (HF) or all-cause mortality events. The second outcome included non-fatal myocardial infarction (MI), non-fatal stroke, changes of left ventricular ejection fraction (LVEF) and 6-minute walking distance (6MWD) from baseline to 24 months after randomization. Results Finally, 138 patients of the ALA group and 139 patients of the placebo group attained the primary outcome. Hospitalization for HF or all-cause mortality events occurred in 32 patients (23.2%) of the ALA group and in 40 patients (28.8%) of the placebo group (HR＝0.753, 95%CI 0.473-1.198, P＝0.231; Figure 1A-1C). The absolute risk reduction (ARR) was 5.6%, the relative risk reduction (RRR) associated with ALA therapy was approximately 19.4% compared to placebo, corresponding to a number needed to treat (NNT) of 18 patients to prevent one event. In the secondary outcome analysis, the composite outcome of the major adverse cardiovascular events (MACE) including the hospitalization for HF, all-cause mortality events, non-fatal MI or non-fatal stroke occurred in 35 patients (25.4%) in the ALA group and 47 patients (33.8%) in the placebo group (HR＝0.685, 95%CI 0.442-1.062, P＝0.091; Figure 1D). Moreover, greater improvement in LVEF (β＝3.20, 95%CI 1.14-5.23, P＝0.002) and 6MWD (β＝31.7, 95%CI 8.3-54.7, P＝0.008) from baseline to 24 months after randomization were observed in the ALA group as compared to the placebo group. There were no differences in adverse events between the study groups. Conclusions These results show potential long-term beneficial effects of adding ALA to IHF patients. ALA could significantly improve LVEF and 6MWD compared to the placebo group in IHF patients.

Aim To study the therapeutic effect of al-licin on senna-induced diarrhea in mice and to explore the underlying mechanism.Methods Forty-eight C57BL/6J mice were randomly divided into six groups:control,model,loperamide positive control group(2 mg·kg-1),allicin low-dose group(6 mg·kg-1),allicin medium-dose group(12 mg·kg-1)and allicin high-dose group(18 mg·kg-1).Except for the con-trol group,the diarrhea model was induced in the other groups by intragastric administration of senna leaf ex-tract.After drug administration,several diarrhea indi-ces were measured:the rate of loose stools,diarrhea index,accumulated frequency of loose stools at differ-ent time points within 5 hours,and small intestine pro-pelling rate.Serum levels of TNF-α and IL-6 were de-tected by ELISA.Serum NO content was determined u-sing the Griess method.The activities of SOD and CAT,as well as MDA content in the ileum and colon,were measured.The pathological changes and the ex-pression of mRNA related to intestinal barrier proteins in the ileum and colon were evaluated using HE stai-ning and RT-qPCR.Results Allicin improved diar-rhea symptoms in mice induced by senna leaf.It re-duced the rate of loose stools,diarrhea index,cumula-tive number of loose stools in five hours,and the intes-tinal propulsion rate.Allicin also protected the intesti-nal mucosa,decreased serum TNF-α and IL-6 levels,and lowered MDA content in the intestines.It in-creased serum NO levels and enhanced SOD and CAT activities in the intestines.Additionally,allicin upreg-ulated the mRNA expression of AQP1,AQP4,and ZO-1 in intestinal tissues.Conclusions Allicin has a significant therapeutic effect on senna-induced diarrhea in mice.The underlying molecular mechanisms may involve anti-inflammatory and antioxidant effects,in-creased NO content,and upregulation of mRNA ex-pression of aquaporins and tight-junction proteins.

BACKGROUND:Bone morphogenetic protein 2 is vital in embryonic development,bone formation,and regeneration,but its high-dose application is linked to cancer.Bone morphogenetic protein 2 osteogenic peptide L20 reduces adverse effects like cancer and boosts bone tissue regeneration.OBJECTIVE:To graft bone morphogenetic protein 2 active peptide segments onto mesopores and surfaces through a peptide mimicry strategy inspired by oysters,and explore its impact on osteogenic properties of tissue-engineered bone.METHODS:(1)Mesoporous bioactive glass was synthesized using a template method.Bone morphogenetic protein 2 osteogenic peptide L20 was loaded onto mesoporous bioactive glass using a one-step synthesis method to characterize the morphology and in vitro sustained release properties of mesoporous active glass nanoparticles loaded with bone morphogenetic protein 2 osteogenic active peptide L20.(2)Bone marrow mesenchymal stem cells were isolated and extracted from SD rats.After two generations,they were co-cultured with PBS(blank group),mesoporous bioactive glass nanoparticles(control group),and mesoporous bioactive glass nanoparticles loaded with bone morphogenetic protein 2 osteogenic active peptide L20(experimental group).Cell live/dead fluorescence staining and CCK-8 assay were used to detect cytotoxicity and cell proliferation.Scanning electron microscopy was used to observe cell adhesion.After osteogenic induction and differentiation,alkaline phosphatase staining,Alizarin red S staining,and osteogenesis-related gene expression were detected.(3)Fifteen SD rats were selected to establish bilateral femoral condyle defect models and divided into three groups using a random number table method:the blank group(n=5)was not implanted with any material;the control group(n=5)was implanted with mesoporous bioactive glass nanoparticles,and the experimental group(n=5)was implanted with mesoporous bioactive glass nanoparticles loaded with bone morphogenetic protein 2 osteogenic active peptide L20.Eight weeks after surgery,femoral Micro-CT scanning and tissue morphology observation were performed.RESULTS AND CONCLUSION:(1)Scanning electron microscopy showed that the mesoporous bioactive glass nanoparticles loaded with bone morphogenetic protein 2 osteogenic active peptide L20 were spherical and monodisperse particles.Transmission electron microscopy showed their porous structure with an average particle size of(268.10±0.58)nm,which could release L20 in vitro.(2)Mesoporous bioglass nanoparticles loaded with bone morphogenetic protein 2 osteogenic active peptide L20 were non-cytotoxic and could promote the proliferation and adhesion of bone marrow mesenchymal stem cells.Compared with the blank group and the control group,the alkaline phosphatase activity and extracellular matrix mineralization capacity of the experimental group were increased(P<0.05),and the mRNA expression levels of alkaline phosphatase,Runx2,and osteocalcin were increased(P<0.05).(3)The results of femoral Micro-CT scanning showed that compared with the blank group and the control group,the new bone mass and bone density of the experimental group were increased(P<0.05).The results of hematoxylin-eosin and Masson staining showed that compared with the blank group and the control group,the new bone formation and collagen fibers of the experimental group were increased.(4)These findings indicate that mesoporous bioactive glass loaded with bone morphogenetic protein 2 active peptide L20 exhibits excellent biocompatibility and in vitro and in vivo osteogenic properties,promoting regeneration and repair of SD rat femoral condyle defects.

Aim To study the therapeutic effect of al-licin on senna-induced diarrhea in mice and to explore the underlying mechanism.Methods Forty-eight C57BL/6J mice were randomly divided into six groups:control,model,loperamide positive control group(2 mg·kg-1),allicin low-dose group(6 mg·kg-1),allicin medium-dose group(12 mg·kg-1)and allicin high-dose group(18 mg·kg-1).Except for the con-trol group,the diarrhea model was induced in the other groups by intragastric administration of senna leaf ex-tract.After drug administration,several diarrhea indi-ces were measured:the rate of loose stools,diarrhea index,accumulated frequency of loose stools at differ-ent time points within 5 hours,and small intestine pro-pelling rate.Serum levels of TNF-α and IL-6 were de-tected by ELISA.Serum NO content was determined u-sing the Griess method.The activities of SOD and CAT,as well as MDA content in the ileum and colon,were measured.The pathological changes and the ex-pression of mRNA related to intestinal barrier proteins in the ileum and colon were evaluated using HE stai-ning and RT-qPCR.Results Allicin improved diar-rhea symptoms in mice induced by senna leaf.It re-duced the rate of loose stools,diarrhea index,cumula-tive number of loose stools in five hours,and the intes-tinal propulsion rate.Allicin also protected the intesti-nal mucosa,decreased serum TNF-α and IL-6 levels,and lowered MDA content in the intestines.It in-creased serum NO levels and enhanced SOD and CAT activities in the intestines.Additionally,allicin upreg-ulated the mRNA expression of AQP1,AQP4,and ZO-1 in intestinal tissues.Conclusions Allicin has a significant therapeutic effect on senna-induced diarrhea in mice.The underlying molecular mechanisms may involve anti-inflammatory and antioxidant effects,in-creased NO content,and upregulation of mRNA ex-pression of aquaporins and tight-junction proteins.

Objective:To systematically evaluate the clinical outcomes of hemihepatic inflow occlusion(HHO)and total hepatic inflow occlusion(THO)in liver resection surgery.Method:Re-trieve the Cochrane Library,PubMed,EMbase,Ovid,Web of Science,CNKI,and WanFang Data databases,and search for journal articles published from January 1,2000 to January 31,2023,on randomized controlled trials(RCTs)comparing the effects of HHO and THO in liver resection.At the same time,two researchers independently screened literature based on inclusion and exclusion cri-teria,conducted meta-analysis using RevMan 5.4 and State17PM software.Result:Fifteen RCTs were ultimately included,including 624 patients.The meta-analysis results showed that HHO re-duced serum AST(MD=-104.75,95％CI:-134.45-75.06,P＜0.05),ALT(MD=-155.37,95％CI:-182.90-127.85,P＜0.05),and TBIL(MD=-6.28,95％CI:-8.07-4.48,P＜0.05)on postoperative days 1,3,and 7 Compared to THO,the elevation of blood levels and reduction of intraoperative bleeding(MD=-66.21,95％CI:-116.49-15.94,P＜0.05)were superior.THO is superior to HHO in shortening surgical time(MD=13.94,95％CI:4.77-23.12,P＜0.05).There was no significant differ-ence between the two methods in hospital stay,hospital death rate and complication rate(P＞0.05).Conclusion:compared with THO,the application of HHO in hepatectomy has less damage to liver function and less intraoperative bleeding,but the two methods have the similar effect in terms of hospitalization time,hospital mortality rate and complication rate.

AIM To study the chemical constituents from the branches and leaves of Toona ciliata Roem.var.pubescens(Franch.)Hand-Mazz.and their antitumor activities.METHODS The compounds were isolated and purified by silica gel,RP-18 reverse phase silica gel and semi-preparative HPLC,the structures of compounds were identified by physicochemical properties and spectral data.The antitumor activities were determined by MTT method.RESULTS Fifteen compounds were isolated and identified as toonaolide D(1),toonaciliatin E(2),bourjotinolone A(3),(21R,23R)-epoxy-21α-ethoxy-24S,25-dihydroxyapotirucalla-7-en-3-one(4),(Z)-toonasterone C(5),(E)-toonasterone(6),3-epi-dyscusin C(7),(Z)-aglawone(8),(E)-volkendousin(9),8(14),15-isopimaradiene-2α,3α,19-triol(10),(-)-loliolide(11),cyclohexenone(12),pubinernoid A(13),quercetin-3-O-(4″-methoxy)-α-L-rahmnopyranosyl(14),5-hydroxymethyl-2-furancarboxaldehyde(15).The IC50 values of compounds 3 and 4 on K562 cells were 54.2 and 47.3 μmol/L,respectively,and the IC50 values on HEL cells were 47.3 and 61.1 μmol/L,respectively.CONCLUTION Compounds 4,7,10 and 11 are isolated from Toona genus for the first time,and compounds 2,15 are first isolated from this plant.Compounds 3 and 4 show weak antitumor activities.