Objective To investigate the effects of Buyang Huanwu Decoction on nerve regeneration after cerebral ischemia in mice based on Cav-1/Notch1/Hes1 signaling pathway.Methods Wild(WT)and Cav-1-/-(KO)male mice were randomly divided into sham-operation group,model group and Buyang Huanwu Decoction group,with 10 mice in each group.The middle cerebral artery occlusion method was used to construct the mouse cerebral ischemia model.After modeling,5-ethynyl-2'-deoxyuridine(EdU)was injected intraperitoneally every 7 days for 3 times(with an interval of 8 hours).Buyang Huanwu Decoction group was given 18.5 g/kg Buyang Huanwu Decoction by gavage daily,while the sham-operation group and model group were given distilled water of equal volume by gavage for 21 consecutive days.The neurological function of mice was evaluated using modified neurological severity score,brain tissue morphology was observed through HE staining,nerve regeneration in the ischemic side injury area was detected using dual immunofluorescence staining,Western blot was used to detect the expressions of Notch1 and Hes1 protein in ischemic cortical area.Results Compared with the same genotype sham-operation group,the neurological function score of the mice in the WT and KO model groups significantly increased(P<0.01),the arrangement of neurons in the ischemic cortical area was disordered,with nucleoli shrinking,marginalization,and even rupture,widened intercellular spaces,intracellular edema and vacuolar changes,the co-localization of EdU/Nestin,EdU/DCX and EdU/NeuN in the ischemic side injury area significantly increased(P<0.01),and the expression of Notch1 and Hes1 protein in ischemic cortical area significantly increased(P<0.01).Compared with the same genotype model group,the neurological function score of the WT and KO Buyang Huanwu Decoction group significantly decreased(P<0.01),the neurons in the ischemic cortical area were arranged neatly and structurally intact,with reduced intercellular space and clear nucleoli,the co-localization of EdU/Nestin,EdU/DCX and EdU/NeuN in the ischemic side injury area was significantly increased(P<0.01),and the expression of Notch1 and Hes1 protein in ischemic cortical area were significantly decreased(P<0.01).Compared with the corresponding WT group,the neurological function scores of mice in the KO model group and KO Buyang Huanwu Decoction group significantly increased(P<0.01),the neurons in the ischemic cortical area had more severe nucleolar condensation,marginalization and rupture,with more vacuolar changes and ischemic injury,the co-localization of EdU/Nestin and EdU/NeuN in the ischemic side injury area was significantly decreased(P<0.01),and the expression of Notch1 and Hes1 proteins in ischemic cortical area significantly increased(P<0.01).Conclusion Buyang Huanwu Decoction can promote nerve regeneration after cerebral ischemia,which may be related to the regulation of Cav-1 thereby inhibition Notch1//Hes1 signaling pathway activity.

BACKGROUND:Troxerutin has been found to have a significant ameliorative effect on brain disorders,but there are fewer studies on the effects of troxerutin on the treatment of cerebral infarction and on neuronal cells. OBJECTIVE:To investigate the mechanism by which troxerutin regulates nuclear factor-κB signaling pathway to reduce brain injury and neuronal apoptosis in cerebral infarction rats. METHODS:Fifty clean grade rats were randomized into healthy group,model group,and troxerutin+nuclear factor-κB agonist group,troxerutin group,and nuclear factor-κB inhibitor group.Except for the healthy group,all other groups were used to establish a rat model of cerebral infarction by arterial ligation.The healthy and model groups were treated once a day with an equal amount of physiological saline by gavage.The troxerutin+nuclear factor-κB agonist group was intervened with 72 mg/kg troxerutin by gavage+20 mg/kg RANK intraperitoneally.The troxerutin group was treated with 72 mg/kg troxerutin by gavage.The nuclear factor κB inhibitor group was intervened intraperitoneally with 120 mg/kg nuclear factor κB inhibitor pyrrolidine disulfiram.Administration in each group was given once a day for 30 continuous days.Zea-longa was used to detect neurological damage in rats,hematoxylin-eosin staining was used to observe pathological changes,TUNEL was used to detect neuronal apoptosis,and immunoblotting and PCR were used to detect the expression of nuclear factor-κB p65 and nuclear factor-κB p50 at protein and mRNA levels,respectively. RESULTS AND CONCLUSION:Compared with the healthy group,the neurological function score,neuronal apoptosis rate,nuclear factor-κB p65,nuclear factor-κB p50 mRNA and protein expression levels were elevated in the model group(P＜0.05).Compared with the model group,the neurological function score,neuronal apoptosis rate,nuclear factor-κB p65 and nuclear factor-κB p50 mRNA and protein expression levels were decreased in the troxerutin+nuclear factor-κB agonist group(P＜0.05).Compared with the troxerutin+nuclear factor-κB agonist group,the neurological function score,neuronal apoptosis rate,nuclear factor-κB p65 and nuclear factor-κB p50 mRNA and protein expression levels were reduced in the troxerutin group and nuclear factor-κB inhibitor group(P＜0.05).In addition,there was no difference between the troxerutin group and the nuclear factor-κB inhibitor group(P＞0.05).In the model group,there was a large number of cytoplasmic vacuolation,obvious edema and necrosis,and a large number of inflammatory cell infiltrations.In the troxerutin+nuclear factor-κB agonist,the swelling of brain tissue was reduced,and reticulate structures and condensed cells were reduced,still with some edema.In the troxerutin group and nuclear factor-κB inhibitor group,brain tissue swelling,neuronal edema degeneration,cytoplasmic vacuolation and neuronal nucleus consolidation were reduced,and the inflammatory cell infiltration was significantly decreased.To conclude,troxrutin can reduce the expression of neurological impairment,inhibit neuronal apoptosis and improve the pathological injury of brain tissue in rats with cerebral infarction,and its mechanism of action may be related to the modulation of nuclear factor-κB expression and related signaling pathways.

Objective:To investigate the efficacy and safety of venetoclax and azacitidine combined with GHA (human granulocyte colony stimulating factor, homoharringtonine and low-dose cytarabine) priming regimen in treatment of patients with relapsed/refractory acute myeloid leukemia.Methods:A retrospective case series study was conducted. Twenty-three patients with relapsed/refractory acute myeloid leukemia (non-acute promyelocytic leukemia) who received treatment with the combination of venetoclax and azacitidine with GHA priming regimen at the Second Affiliated Hospital of Xi'an Jiaotong University from October 2020 to July 2024 were selected, and the treatment efficacy, minimal residual disease (MRD)-negative rate in patients with comprehensive complete remission (cCR) (including complete remission, complete remission with partial hematologic recovery and complete remission with incomplete hematologic recovery) and the adverse reactions were analyzed; patients were followed-up, and their overall survival (OS) was analyzed by using Kaplan-Meier method.Results:The median age of the 23 patients was 60 years (range: 21-79 years), including 10 males and 13 females. The cCR rate for 1 course of treatment was 52.2% (12/23), with 4 cases of MRD negative among cCR patients; 5 cases received 2 courses of treatment, with 3 cases achieving cCR, of which 2 cases were MRD negative; 2 cases received 3 courses of treatment, with 1 case achieving complete remission with incomplete hematologic recovery. Six patients underwent allogeneic hematopoietic stem cell transplantation. The patients were followed up until July 31, 2024, and the median follow-up period was 5.3 months (range: 1.1-41.7 months). Ten cases survived, 12 cases died, 1 case was lost to follow-up, and the median OS time of 23 patients was 7.9 months. The 6-month OS rate was 60.2% (95% CI: 42.7%-84.8%), and the 12-month OS rate was 44.6% (95% CI: 26.8%-74.3%). Common adverse reactions during treatment included infection [69.6% (16/23)], nausea [56.5% (13/23)], febrile neutropenia [52.2% (12/23)], bleeding [52.2% (12/23)], vomiting [34.8% (8/23)], and pneumonia [34.8% (8/23)]. Conclusions:The combination of vinaclotide and azacitidine with GHA priming regimen has certain efficacy and good safety in the treatment of relapsed/refractory acute myeloid leukemia.

Objective·To investigate an optimal severe periodontitis mouse model by comparing two induction methods:simple ligature and ligature combined with injection of Porphyromonas gingivalis lipopolysaccharide(P.g.LPS).Methods·Fifteen C57BL/6 mice were divided into three groups:a healthy control group,a simple ligature-induced periodontitis group,and a ligature combined with P.g.LPS injection-induced periodontitis group.After 14 d,the following evaluations were conducted:tooth mobility and probing depth under a stereomicroscope;alveolar bone resorption[bone volume fraction,bone mineral density,the distance from the cemento-enamel junction(CEJ)to the alveolar bone crest(ABC),and the area between CEJ and ABC]analyzed via micro computed tomography(Micro-CT)and stereomicroscopic examination.The serum levels of inflammatory cytokines interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α)were assessed by enzyme-linked immunosorbent assay(ELISA).Results·Compared with the simple ligature group,mice in the ligature with P.g.LPS injection group exhibited significantly increased tooth mobility[(2.20±0.45)vs(1.40±0.55)]and probing depth[(1.05±0.21)mm vs(0.58±0.39)mm],with statistically significant differences(P＜0.05).The ligature with P.g.LPS injection group also demonstrated significantly reduced bone volume fraction[(16.44%±3.35%)vs(28.97%±7.90%)]and bone mineral density[(0.42±0.04)g/cm3 vs(0.55±0.08)g/cm3],as well as increased distance from CEJ to ABC[(0.88±0.03)mm vs(0.74±0.12)mm]and area between CEJ and ABC[(0.34±0.01)mm2 vs(0.30±0.02)mm2],all with statistically significant differences(all P＜0.05).Additionally,serum levels of TNF-α and IL-1β were significantly elevated in the ligature with P.g.LPS injection group compared to the simple ligature group(both P＜0.05).Conclusion·The method of ligature combined with continuous P.g.LPS injection is more effective for constructing a severe periodontitis mouse model,making it suitable for studying the progression and treatment of severe periodontitis.

Objective To investigate the effects of Buyang Huanwu Decoction on nerve regeneration after cerebral ischemia in mice based on Cav-1/Notch1/Hes1 signaling pathway.Methods Wild(WT)and Cav-1-/-(KO)male mice were randomly divided into sham-operation group,model group and Buyang Huanwu Decoction group,with 10 mice in each group.The middle cerebral artery occlusion method was used to construct the mouse cerebral ischemia model.After modeling,5-ethynyl-2'-deoxyuridine(EdU)was injected intraperitoneally every 7 days for 3 times(with an interval of 8 hours).Buyang Huanwu Decoction group was given 18.5 g/kg Buyang Huanwu Decoction by gavage daily,while the sham-operation group and model group were given distilled water of equal volume by gavage for 21 consecutive days.The neurological function of mice was evaluated using modified neurological severity score,brain tissue morphology was observed through HE staining,nerve regeneration in the ischemic side injury area was detected using dual immunofluorescence staining,Western blot was used to detect the expressions of Notch1 and Hes1 protein in ischemic cortical area.Results Compared with the same genotype sham-operation group,the neurological function score of the mice in the WT and KO model groups significantly increased(P<0.01),the arrangement of neurons in the ischemic cortical area was disordered,with nucleoli shrinking,marginalization,and even rupture,widened intercellular spaces,intracellular edema and vacuolar changes,the co-localization of EdU/Nestin,EdU/DCX and EdU/NeuN in the ischemic side injury area significantly increased(P<0.01),and the expression of Notch1 and Hes1 protein in ischemic cortical area significantly increased(P<0.01).Compared with the same genotype model group,the neurological function score of the WT and KO Buyang Huanwu Decoction group significantly decreased(P<0.01),the neurons in the ischemic cortical area were arranged neatly and structurally intact,with reduced intercellular space and clear nucleoli,the co-localization of EdU/Nestin,EdU/DCX and EdU/NeuN in the ischemic side injury area was significantly increased(P<0.01),and the expression of Notch1 and Hes1 protein in ischemic cortical area were significantly decreased(P<0.01).Compared with the corresponding WT group,the neurological function scores of mice in the KO model group and KO Buyang Huanwu Decoction group significantly increased(P<0.01),the neurons in the ischemic cortical area had more severe nucleolar condensation,marginalization and rupture,with more vacuolar changes and ischemic injury,the co-localization of EdU/Nestin and EdU/NeuN in the ischemic side injury area was significantly decreased(P<0.01),and the expression of Notch1 and Hes1 proteins in ischemic cortical area significantly increased(P<0.01).Conclusion Buyang Huanwu Decoction can promote nerve regeneration after cerebral ischemia,which may be related to the regulation of Cav-1 thereby inhibition Notch1//Hes1 signaling pathway activity.

Objective·To investigate an optimal severe periodontitis mouse model by comparing two induction methods:simple ligature and ligature combined with injection of Porphyromonas gingivalis lipopolysaccharide(P.g.LPS).Methods·Fifteen C57BL/6 mice were divided into three groups:a healthy control group,a simple ligature-induced periodontitis group,and a ligature combined with P.g.LPS injection-induced periodontitis group.After 14 d,the following evaluations were conducted:tooth mobility and probing depth under a stereomicroscope;alveolar bone resorption[bone volume fraction,bone mineral density,the distance from the cemento-enamel junction(CEJ)to the alveolar bone crest(ABC),and the area between CEJ and ABC]analyzed via micro computed tomography(Micro-CT)and stereomicroscopic examination.The serum levels of inflammatory cytokines interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α)were assessed by enzyme-linked immunosorbent assay(ELISA).Results·Compared with the simple ligature group,mice in the ligature with P.g.LPS injection group exhibited significantly increased tooth mobility[(2.20±0.45)vs(1.40±0.55)]and probing depth[(1.05±0.21)mm vs(0.58±0.39)mm],with statistically significant differences(P＜0.05).The ligature with P.g.LPS injection group also demonstrated significantly reduced bone volume fraction[(16.44%±3.35%)vs(28.97%±7.90%)]and bone mineral density[(0.42±0.04)g/cm3 vs(0.55±0.08)g/cm3],as well as increased distance from CEJ to ABC[(0.88±0.03)mm vs(0.74±0.12)mm]and area between CEJ and ABC[(0.34±0.01)mm2 vs(0.30±0.02)mm2],all with statistically significant differences(all P＜0.05).Additionally,serum levels of TNF-α and IL-1β were significantly elevated in the ligature with P.g.LPS injection group compared to the simple ligature group(both P＜0.05).Conclusion·The method of ligature combined with continuous P.g.LPS injection is more effective for constructing a severe periodontitis mouse model,making it suitable for studying the progression and treatment of severe periodontitis.

Objective To explore the effects of Buyang Huanwu Decoction on neuronal cell apoptosis after cerebral ischemia based on mediating Cav1 in regulating Wnt pathway.Methods Male wild-type(WT)and Cav1-/-(KO)C57BL/6 mice were randomly divided into sham-operation group,model group and Buyang Huanwu Decoction group(18.5 g/kg).Cerebral ischemia model was prepared using middle cerebral artery occlusion method,and drug intervention was given for 14 days.Neurobehavioral score was performed,HE staining was used to observe the morphology of ischemic cortical area of brain tissue,TUNEL staining was used to detect neuronal apoptosis in ischemic cortical area,immunohistochemistry was used to detect the expressions of apoptosis related proteins and Wnt1,glycogen synthase kinase 3β(GSK3β)and β-catenin protein in ischemic cortical area.Results Compared with the same genotype sham-operation group,the neurobehavioral score of the model group mice significantly increased,neuronal cells in the ischemic cortical area showed vacuolar changes,with nuclear condensation and widened intercellular spaces,the apoptosis rate of nerve cells significantly increased,with increased expressions of Bax,GSK3β and decreased expressions of Bcl-2,Wnt1 and β-catenin(P<0.01).Compared with the same genotype model group,the neurobehavioral score of mice in Buyang Huanwu Decoction group were significantly decreased,the pathological damage of the ischemic cortical area improved,the apoptosis rate of nerve cells decreased,the expressions of Bax and GSK3β decreased,and the expressions of Bcl-2,Wnt1 and β-catenin increased(P<0.01).Compared with the WT model group,the KO model group showed an increase in neurobehavioral score,aggravated damage in ischemic cortical area,significantly increased neuronal apoptosis rate,and increased expression of GSK3β(P<0.05).Compared with the WT Buyang Huanwu Decoction group,the KO Buyang Huanwu Decoction group showed an increase in neurobehavioral score,aggravated damage in ischemic cortical area,significantly increased neuronal apoptosis rate,increased expressions of Bax and GSK3β,and decreased expressions of Bcl-2,Wnt1 and β-catenin(P<0.01).Conclusion Buyang Huanwu Decoction can inhibit neuronal cell apoptosis after cerebral ischemia,and its mechanism may be related to regulating the expressions of apoptosis-related proteins by mediating Cav1 to regulate the Wnt signaling pathway.

Objective To explore the mechanism of Buyang Huanwu Decoction against cerebral ischemia-reperfusion injury in rats by regulating lipid metabolism through the cAMP/PKA/PPARγ pathway.Methods 60 rats were randomly divided into sham operation group(Sham),model group(Model),Buyang Huanwu Decoction low-dose group(BHD-L),Buyang Huanwu Decoction medium-dose group(BHD-M),Buyang Huanwu Decoction high-dose group(BHD-H)and Butylphthalide group(NBP).The cerebral ischemia-reperfusion model was prepared by transient middle cerebral artery embolization.The BHD low-,medium-and high-groups were given different doses of Buyang Huanwu Decoction(6.413,12.825,25.65 g·kg-1)by intragastric administration.The NBP group was administered with Butylphthalide(54 mg·kg-1).The sham operation group and the model group were administered with an equal volume of distilled water,all given for 14 days.The rats were subjected to neurobehavioral scoring.HE staining was used to observe brain pathological changes,and the kit was used to detect the levels of phosphocholine(PC),phosphatidylethanolamine(PE),diacylglycerol(DAG),and free fatty acid(FFA)on the ischemic side.RT-qPCR and Western Blot were applied to detect the mRNA and protein expressions of cyclic adenosine monophosphate(cAMP),protein kinase A(PKA),and peroxisome proliferator-activated receptor γ(PPARγ).Results Compared with the sham group,the neurological deficit score was significantly increased(P＜0.01),pathomorphological damage in ischemic cortex was found,the contents of PC and PE were reduced,the contents of DAG and FFA were increased(P＜0.01),and cAMP mRNA expression increased(P＜0.05)in the model group.Compared with the model group,the neurological deficit score of the BHD-L group was decreased(P＜0.05),and the neurological deficit score of the BHD-M,BHD-H and NBP groups was significantly decreased(P＜0.01),the cells in each treatment group were regularly arranged,the intercellular spaces were reduced,and the normal cells were increased.PC and PE were significantly increased,DAG and FFA were significantly decreased(P＜0.01)in the BHD-M,BHD-H and NBP groups.PC was increased,FFA and DAG were decreased in the BHD-L group(P＜0.05,P＜0.01).The mRNA level of PPARγ was increased in the BHD-L group(P＜0.05),and the mRNA and protein levels of cAMP,PKA,and PPARγ were increased in the other treatment groups(P＜0.05,P＜0.01).Conclusion Buyang Huanwu Decoction has a neuroprotective effect on cerebral ischemia-reperfusion injury rats,and its mechanism may be related to regulating the expression of key factors in the cAMP/PKA/PPARγ signaling pathway and lipid metabolism.

Objective:To analyze the application and regularity of acupoint selection of Sanyinjiao (SP 6) based on data mining.Methods:Search for literatures in CNKI, Wanfang, VIP, and Pubmed, the clinical researches of acupuncture on Sanyinjiao (SP 6) point were selected, according to the inclusion and exclusion criteria, and the retrieval period was from database construction to September 30th, 2021. Excel 2016, SPSS Statistics 25.0, SPSS Modeler 18.0 were used to perform descriptive analysis, association analysis and cluster analysis.Results:After literature screening, a total of 261 literatures were included, involving 73 kinds of diseases, mainly including mental and behavioral disorders, genitourinary diseases, endocrine and nutritional metabolism diseases and nervous system diseases. The most frequently used acupoints in Sanyinjiao (SP 6) compatibility are Zusanli (ST 36), Baihui (GV 20), Guanyuan (CV 4) and Taichong (LR 3), most of which focus on stomach meridian, conception channel, governor channel and bladder meridian. Seven categories were extracted among high-frequency acupoints by cluster analysis. The association rule analysis showed that the commonly used combination of Sanyinjiao (SP 6) were Zusanli (ST 36)-Sanyinjiao (SP 6), Baihui (GV 20)-Sanyinjiao (SP 6), and Guanyuan (CV 4)-Sanyinjiao (SP 6).Conclusions:Sanyinjiao (SP 6) is widely used in clinical application, and it is always compatible with stomach meridian, conception vessel, governor channel acupoints, especially those acupoints on the outer and inner meridians and the upper and lower parts. Sanyinjiao (SP 6) combined with other acupoints can treat diseases of multiple systems, such as insomnia, stroke, anxiety and depression, dysmenorrhea, infertility, etc. Clustering and association analysis found the core compatibility law of Sanyinjiao (SP 6), which can be used as a reference for clinical acupoint selection.

Objective:To investigate the value of combined detection of neutrophil gelatinase-associated lipocalin(NGAL), cystatin C(CysC)and heme oxygenase-1(HO-1)in the early diagnostic of acute kidney injury(AKI)caused by acute paraquat poisoning(APP)in elderly patients.Methods:One hundred and two elderly APP patients admitted to the emergency department of our hospital from May 2015 to June 2019 were assigned to the observation group, and 50 patients who took physical examinations served as the control group.The observation group was divided into the AKI sub-group(n=59)and the non-AKI sub-group(n=43)based on whether AKI occurred within 72 h of admission.Serum levels of NGAL, CysC, HO-1 and creatinine(Scr)were detected in all APP patients at 0 h(admission), 12 h, 24 h, 48 h and 72 h. Measurements of the same parameters were made on the day of physical examination for the control group.The correlations of serum levels of NGAL, CysC and HO-1 with the occurrence of AKI were analyzed.Relative operating characteristic curve(ROC)was drawn to analyze the diagnostic value of NGAL, CysC, HO-1 and the combination of the three for the early diagnosis of renal injury in APP patients.Results:Serum levels of NGAL, CysC, HO-1 and Scr at admission showed no significant difference between the AKI sub-group, non-AKI sub-group and control group( P>0.05). After admission, all the parameters showed an upward trend in the observation group.Serum levels of NGAL, CysC and HO-1 at 12 h after admission( P<0.05)and Scr levels at 72 h after admission( P<0.05)were significantly different between the AKI sub-group and the non-AKI sub-group.Correlation analysis showed that serum NGAL, CysC and HO-1 levels were positively correlated with the occurrence of AKI in APP patients at 12 h, 24 h, 48 h and 72 h after admission, with the best correlation at 48 h after admission(NGAL: r=0.203, 0.545, 0.707 and 0.560, P<0.05; CysC: r=0.242, 0.340, 0.754 and 0.467, P<0.05; HO-1: r=0.249, 0.536, 0.677 and 0.509, P<0.05). The area under ROC curve predicted by NGAL, CysC, HO-1, Scr and NGAL+ CysC+ HO-1 for AKI at 48 h after admission was 0.777, 0.718, 0.888, 0.602 and 0.969, respectively. Conclusions:Serum levels of NGAL, CysC and HO-1 are significantly elevated at 12 h after admission in elderly APP patients, and reach the peak at 48 h after admission.Each of them can give an earlier diagnosis for AKI than Scr, and the combination of the three provides a higher diagnostic accuracy for AKI.