OBJECTIVE To explore the improvine mechanism of Yifei xuanfei jiangzhuo formula （YFXF） on vascular dementia （VAD） model rats based on the growth factor receptor-bound protein 2 （GRB2）/extracellular signal-regulated kinase （ERK）/cardiolipin synthase 1 （CRLS1） pathway. METHODS VAD rat model was established by permanent bilateral common carotid artery ligation. Forty-eight successfully modeled rats were randomly divided into the model group （normal saline）， donepezil hydrochloride group （positive control group， 0.2 g/kg）， and YFXF low- and high-dose groups （12.18 and 24.36 g/kg， calculated based on the total amount of crude drug）， respectively. In addition， a sham operation group （normal saline） was set up. There were 12 rats in each group. Daily intragastric administration of drug or normal saline was performed for 30 consecutive days. After the last administration， the spatial cognitive ability of the rats was evaluated， the pathological morphology of the hippocampus was observed， the contents of tumor necrosis factor-α （TNF-α） and interleukin-4 （IL-4） in serum were detected， the expression levels of GRB2/ERK/CRLS1 pathway-related proteins and the mRNA levels of GRB2， CRLS1， NADH dehydrogenase subunit 1（ND1）， Tafazzin （TAZ）， phospholipid scramblase 3（PLSCR3） and the ATP content in hippocampal tissue were measured. RESULTS Compared with the sham operation group， the escape latency of rats in the model group was significantly prolonged （ P ＜0.05）， and the number of crossing platform was significantly reduced （ P ＜0.05）， while the number of pyramidal cells and Nissl bodies in the hippocampus decreased sharply； the content of TNF-α in serum was significantly increased （ P ＜0.05）， and the content of IL-4 was significantly decreased （ P ＜0.05）； the expression levels of GRB2 and CRLS1 proteins， the phosphorylation level of ERK protein， the relative expression levels of GRB2， CRLS1，ND1， TAZ， and PLSCR3 mRNA， and the content of ATP in hippocampal tissue were significantly decreased （ P ＜0.05）. Compared with the model group， the above pathological changes in the hippocampal tissue of each administration group were alleviated， and the quantitative indicators were significantly restored （ P ＜0.05）. CONCLUSIONS YFXF may improve hippocampal neuron injury in VAD rats by activating the GRB2/ERK/CRLS1 pathway, maintaining cardiolipin homeostasis， and improving mitochondrial energy metabolism.

AIM:To investigate the mechanism by which the Z-DNA-binding protein 1(ZBP1)/receptor-in-teracting protein kinase 1(RIPK1)/mixed lineage kinase domain-like protein(MLKL)pathway modulates the necroptosis of neurons in a mouse model of Alzheimer disease(AD).METHODS:Thirty mice were randomly divided into three groups:normal control(NC)group,APP/PS1 model(MOD)group,and necroptosis inhibitor necrostatin-1(Nec-1)group,each with 10 mice.The learning and memory capacities of mice were assessed using the Morris water maze assays.The pathological morphology of hippocampal tissue was examined based on the HE staining assay.The expression levels of tumor necrosis factor-α(TNF-α)and interleukin-10(IL-10)in serum samples were measured by ELISA.The phosphory-lation of amyloid precursor protein(APP),Tau protein and the expression levels of proteins related to the ZBP1/RIPK1/MLKL pathway in hippocampal tissue were measured by Western blot.Immunofluorescence analysis was performed to de-tect the positive expression of p-RIPK1,while the mRNA level of ZBP1 was measured by RT-qPCR.RESULTS:Com-pared with NC group,the escape latency of mice in the MOD group was significantly longer(P<0.05),the number of crossing platforms was reduced(P<0.05),and the arrangement of hippocampal neurons was disordered accompanied with nuclear condensation.The concentration of TNF-α in serum was increased,whereas the concentration level of IL-10 was decreased(P<0.05).The expression levels of APP,p-Ttau and ZBP1 proteins in the hippocampal tissue and the ratios of p-RIPK1/RIPK1,p-RIPK3/RIPK3 and p-MLKL/MLKL were significantly upregulated(P<0.05).Similarly,the positive expression level of p-RIPK1 and the mRNA level of ZBP1 in hippocampal tissue were significantly upregulated(P<0.05).Compared with the MOD group,the cognitive function of AD mice,pathological damage of hippocampal tissue,and the levels of TNF-α and IL-10 in serum were reversed in the Nec-1 group(P<0.05).Moreover,the Nec-1 treatment signifi-cantly downregulated the expression levels of the above proteins(P<0.05),and the positive expression of p-RIPK1 and the mRNA level of ZBP1 were significantly decreased(P<0.05).CONCLUSION:The ZBP1/RIPK1/MLKL pathway is involved in the occurrence of neuronal necroptosis and the pathological process of AD mice.Inhibition of this pathway sig-nificantly ameliorates cognitive dysfunction and neuroinflammatory responses in AD mice.

OBJECTIVE To investigate the effects and mechanism of Wenpi tongluo kaiqiao formula （WPTL） against neuronal necroptosis in Alzheimer’s disease （AD） mice based on the Z-DNA binding protein 1 （ZBP1）/mixed lineage kinase domain-like protein （MLKL） signaling pathway. METHODS Forty APP/PS1 transgenic AD mice were randomly divided into model group， WPTL low-dose （WPTL-L） group （10.4 g/kg， calculated by the raw medicine）， WPTL high-dose （WPTL-H） group （20.8 g/kg， calculated by the raw medicine） and donepezil hydrochloride group （3 mg/kg）， with 10 mice in each group； another 10 C57BL/6J mice were selected as normal control group. Intragastric administration， once a day， for 30 consecutive days. Twenty-four hours after the last administration， Morris water maze test was performed to evaluate learning and memory abilities； the pathological morphology of hippocampal tissues was observed； the serum levels of tumor necrosis factor-α （TNF-α） and interleukin-4 （IL-4） were determined； the expressions of amyloid precursor protein （APP）， Tau protein， and ZBP1/MLKL signaling pathway-related proteins in hippocampal tissues were detected； the positive expression of phosphorylated receptor-interacting protein kinase 3 （p-RIPK3） in the neurons of hippocampal tissues and mRNA expression of ZBP1 were measured in hippocampal tissues. RESULTS Compared with normal control group， the escape latency of mice in model group was prolonged significantly on day 3 to 5 （P＜0.05）， the times of crossing platform reduced significantly （P＜0.05）， and obvious pathological changes were observed in the hippocampal tissue. The level of TNF- α， the expressions of APP， p-Tau and ZBP1， the phosphorylation levels of RIPK1， RIPK3 and MLKL， the fluorescence intensity of p-RIPK3 as well as the mRNA expression of ZBP1 were significantly increased （P＜0.05）， while the serum level of IL-4 was decreased significantly （P＜0.05）. Compared with model group， above indexes were reversed significantly in administration groups （P＜0.05）， and pathological damage of hippocampal tissue was alleviated. CONCLUSIONS WPTL can inhibit the ZBP1/MLKL signaling pathway， reduce neuronal necroptosis in AD mice， and inhibit inflammatory responses， thereby improving learning and spatial memory abilities in AD mice.

OBJECTIVE To investigate the intervention effect and its potential mechanism of Yifei xuanfei jiangzhuo formula by inhibiting mitochondrial fission in a vascular dementia(VaD)model rats.METHODS VaD rat model was established by bilateral common carotid artery ligation.The experimental animals were randomly divided into sham operation group(SHAM),model group(MOD),Yifei xuanfei jiangzhuo formula low-dose group(YFXF-L),Yifei xuanfei jiangzhuo formula high-dose group(YFXF-H),and Donepezil hydrochloride group(positive control),with 9 animals in each group.After 30 days of intervention,the spatial learning memory ability was assessed by Morris water maze experiment;HE staining was used to observe histopathological changes in CA1 area of hippocampus;ELISA was used to detect the levels of serum inflammatory factors[interleukin-1β(IL-1β)and IL-4];Western blot was used to detect the expressions of heat shock protein 90(HSP90)/mixed lineage kinase domain-like protein(MLKL)/dynamin-related protein 1(Drp1)pathway-related proteins,mitochondrial fusion proteins(MFN1,MFN2),and adenosine triphosphate synthase 5A(ATP5A)in hippocampal tissues.The immunohistochemistry was used to detect the level of phosphorylated MLKL(p-MLKL);real-time fluorescence quantitative PCR was adopted to detect mRNA expressions of HSP90,MFN1,MFN2 and ATP5A.RESULTS Compared with SHAM group,the escape latency of rats in the MOD group was significantly prolonged,the number of crossing the platform was significantly reduced,and the hippocampal tissues showed typical neuronal damage characteristics,the positive expression level of p-MLKL and the serum level of IL-1β significantly increased,while the serum level of IL-4 significantly decreased,the protein and mRNA expression of HSP90,as well as the protein expressions of p-MLKL/MLKL and p-Drp1(Ser616)/Drp1 were all significantly increased in hippocampal tissue,the protein and mRNA expressions of MFN1,MFN2 and ATP5A,and protein expression of p-Drp1(Ser637)/Drp1 were all significantly decreased(P＜0.05).After the intervention of Yifei xuanfei jiangzhuo formula,above indicators in each treatment group were all significantly reversed(P＜0.05).CONCLUSIONS Yifei xuanfei jiangzhuo formula may alleviate neuronal damage and neuroinflammatory responses in VaD rats by regulating the HSP90/MLKL/Drp1 signaling pathway,inhibiting mitochondrial fission,thereby maintaining mitochondrial dynamic balance and improving mitochondrial function.

AIM:To investigate the mechanism by which the Z-DNA-binding protein 1(ZBP1)/receptor-in-teracting protein kinase 1(RIPK1)/mixed lineage kinase domain-like protein(MLKL)pathway modulates the necroptosis of neurons in a mouse model of Alzheimer disease(AD).METHODS:Thirty mice were randomly divided into three groups:normal control(NC)group,APP/PS1 model(MOD)group,and necroptosis inhibitor necrostatin-1(Nec-1)group,each with 10 mice.The learning and memory capacities of mice were assessed using the Morris water maze assays.The pathological morphology of hippocampal tissue was examined based on the HE staining assay.The expression levels of tumor necrosis factor-α(TNF-α)and interleukin-10(IL-10)in serum samples were measured by ELISA.The phosphory-lation of amyloid precursor protein(APP),Tau protein and the expression levels of proteins related to the ZBP1/RIPK1/MLKL pathway in hippocampal tissue were measured by Western blot.Immunofluorescence analysis was performed to de-tect the positive expression of p-RIPK1,while the mRNA level of ZBP1 was measured by RT-qPCR.RESULTS:Com-pared with NC group,the escape latency of mice in the MOD group was significantly longer(P<0.05),the number of crossing platforms was reduced(P<0.05),and the arrangement of hippocampal neurons was disordered accompanied with nuclear condensation.The concentration of TNF-α in serum was increased,whereas the concentration level of IL-10 was decreased(P<0.05).The expression levels of APP,p-Ttau and ZBP1 proteins in the hippocampal tissue and the ratios of p-RIPK1/RIPK1,p-RIPK3/RIPK3 and p-MLKL/MLKL were significantly upregulated(P<0.05).Similarly,the positive expression level of p-RIPK1 and the mRNA level of ZBP1 in hippocampal tissue were significantly upregulated(P<0.05).Compared with the MOD group,the cognitive function of AD mice,pathological damage of hippocampal tissue,and the levels of TNF-α and IL-10 in serum were reversed in the Nec-1 group(P<0.05).Moreover,the Nec-1 treatment signifi-cantly downregulated the expression levels of the above proteins(P<0.05),and the positive expression of p-RIPK1 and the mRNA level of ZBP1 were significantly decreased(P<0.05).CONCLUSION:The ZBP1/RIPK1/MLKL pathway is involved in the occurrence of neuronal necroptosis and the pathological process of AD mice.Inhibition of this pathway sig-nificantly ameliorates cognitive dysfunction and neuroinflammatory responses in AD mice.

OBJECTIVE To investigate the mechanism of Yifei xuanfei jiangzhuo formula （YFXF） against vascular dementia （VD）. METHODS The differentially expressed genes of YFXF （YDEGs） were obtained by network pharmacology. High-risk genes were screened from YDEGs by using the nomogram model. The optimal machine learning models in generalized linear， support vector machine， extreme gradient boosting and random forest models were screened based on high-risk genes. VD model rats were established by bilateral common carotid artery occlusion， and were randomly divided into model group and YFXF group （12.18 g/kg， by the total amount of crude drugs）， and sham operation group was established additionally， with 6 rats in each group. The effects of YFXF on behavior （using escape latency and times of crossing platform as indexes）， histopathologic changes of cerebral cortex， and the expression of proteins related to the secreted phosphoprotein 1 （SPP1）/phosphoinositide 3-kinase （PI3K）/protein kinase B （aka Akt） signaling pathway and the mRNA expression of SPP1 in cerebral cortex of VD rats were evaluated. RESULTS A total of 6 YDEGs were obtained， among which SPP1， CCL2， HMOX1 and HSPB1 may be high-risk genes of VD. The generalized linear model based on high-risk genes had the highest prediction accuracy （area under the curve of 0.954）. Compared with the model group， YFXF could significantly shorten the escape latency of VD rats， significantly increase the times of crossing platform （P＜0.05）； improve the pathological damage of cerebral cortex， such as neuronal shrinkage and neuronal necrosis； significantly reduce the expressions of SPP1 protein and mRNA （P＜0.05）， while significantly increase the phosphorylation levels of PI3K and Akt （P＜0.05）. CONCLUSIONS VD high-risk genes SPP1， CCL2， HMOX1 and HSPB1 may be the important targets of YFXF. YFXF may play an anti-VD role by down-regulating the protein and mRNA expressions of SPP1 and activating PI3K/Akt signaling pathway.

Objective To investigate the effect of hyperbaric oxygen (HBO) on malignant biological behavior of ovarian cancer SKOV3 cells and its related mechanism.Methods After the culture of human ovarian cancer cell line SKOV3 was completed,the effects of HBO on the proliferation,apoptosis and tumorigenicity of SKOV3 cells were respectively detected by CCK-8,flow cytometry and Western blotting in the HBO group and the control group,and the effects of HBO on PTEN/PI3K/AKT pathway activation were detected by Western blotting.After PTEN was knocked down,the effects of HBO on the proliferation and apoptosis of SKOV3 cells were detected by the above methods.The inhibition of HBO on the growth of ovarian cancer-bearing mice and the expression of related proteins were detected by tumor-bearing experiment in mice.Results Proliferation of SKOV3 cells in the HBO group was significantly decreased,while apoptosis was significantly improved (P ＜ O.05).In the HBO group,the expression of Bcl-2 was decreased,while the expression of Bax and caspase-3 were increased (P ＜ 0.05).HBO could significantly increase the expression of PTEN protein,and at the same time decrease the expressions of p-PI3K and p-AKT proteins in ovarian cancer SKOV3 cells (P ＜0.05 or P ＜0.01).In addition,the expression of HIF-1 in SKOV3 cells was decreased in the HBO group,however,after knock-down of PTEN,the inhibitory effect of HBO on the expression of HIV-1 in SKOV3 cells was significantly weakened (P ＜ 0.05).The proliferation and apoptosis of SKOV3 cells in the hyperbaric oxygen + si-PTEN group were significantly inhibited (all P ＜ 0.05).Compared with the control group,the tumor size and body mass of mice in the HBO group were significantly decreased (all P ＜ 0.05).The expression levels of HIF-1,p-PI3K,p-AKT and Bcl-2 proteins in the ovarian cancer tissues were decreased,while the expression levels of PTEN,caspase-3 and Bax were increased (all P ＜ 0.05).Conclusion HBO treatment could significantly inhibit malignant biological behavior of ovarian cancer SKOV3 cells,the mechanism of which might be related with the improvement of the hypoxic environment of cells through the regulation of PTEN/PI3K/AKT pathway.

Objective To investigate the effect of hyperbaric oxygen (HBO) on malignant biological behavior of ovarian cancer SKOV3 cells and its related mechanism.Methods After the culture of human ovarian cancer cell line SKOV3 was completed,the effects of HBO on the proliferation,apoptosis and tumorigenicity of SKOV3 cells were respectively detected by CCK-8,flow cytometry and Western blotting in the HBO group and the control group,and the effects of HBO on PTEN/PI3K/AKT pathway activation were detected by Western blotting.After PTEN was knocked down,the effects of HBO on the proliferation and apoptosis of SKOV3 cells were detected by the above methods.The inhibition of HBO on the growth of ovarian cancer-bearing mice and the expression of related proteins were detected by tumor-bearing experiment in mice.Results Proliferation of SKOV3 cells in the HBO group was significantly decreased,while apoptosis was significantly improved (P ＜ O.05).In the HBO group,the expression of Bcl-2 was decreased,while the expression of Bax and caspase-3 were increased (P ＜ 0.05).HBO could significantly increase the expression of PTEN protein,and at the same time decrease the expressions of p-PI3K and p-AKT proteins in ovarian cancer SKOV3 cells (P ＜0.05 or P ＜0.01).In addition,the expression of HIF-1 in SKOV3 cells was decreased in the HBO group,however,after knock-down of PTEN,the inhibitory effect of HBO on the expression of HIV-1 in SKOV3 cells was significantly weakened (P ＜ 0.05).The proliferation and apoptosis of SKOV3 cells in the hyperbaric oxygen + si-PTEN group were significantly inhibited (all P ＜ 0.05).Compared with the control group,the tumor size and body mass of mice in the HBO group were significantly decreased (all P ＜ 0.05).The expression levels of HIF-1,p-PI3K,p-AKT and Bcl-2 proteins in the ovarian cancer tissues were decreased,while the expression levels of PTEN,caspase-3 and Bax were increased (all P ＜ 0.05).Conclusion HBO treatment could significantly inhibit malignant biological behavior of ovarian cancer SKOV3 cells,the mechanism of which might be related with the improvement of the hypoxic environment of cells through the regulation of PTEN/PI3K/AKT pathway.

Vertebrobasilar dolichoectasia(VBD ) can increase the risk of stroke. Significant expansion, elongation and tortuosity of the vertebrobasilar arteries are the main morphology manifestations of VBD. However, there is no consensus on the quantitative imaging assessment of VBD. Many studies showed that some quantitative parameters, such as basilar artery length and bending length, basilar angulation, vertebral tortuosity index can be used for the quantitative imaging assessment of VBD.

Objective To evaluate the impact of different doses of recombinant human B-type natriuretic peptid (rh-BNP) within the dosage of clinical rage on oxygen consumption during acute myocardial infarction (AMI) with heart failure (HF). Methods AMI-HF model of York pig was established by occluding coronary artery with balloon combined with in-jecting microthrombus. Then animals were randomized into rhBNP group and control group. Clinical dose of rhBNP ( 1.5μg/kg bolus followed by a continuous infusion with speed of 0.01, 0.02 and 0.03μg · kg-1 · min-1 for 60 minutes respectively in turn) was administrated in rhBNP group while equal volume of saline was given in the control group. Myocardial oxygen up-take (MOU) was measured by drawing blood from coronary artery and coronary sinus using a catheter. Coronary diameter was determined using quantitative coronary angiography. The observation points were at baseline (T0), instant after the mod-el establishment (T1), 60 min after continuous rhBNP infusion of 0.01, 0.02, 0.03μg·kg-1·min-1 (T2-T4) respectively. Re-sults Compared with the control group, pulmonary capillary wedge pressure, central venous pressure, heart rate, systolic blood pressure and MOU were significantly decreased after rhBNP administration. And cardiac output and coronary diame-ter were obviously increased with addition of rhBNP. There is a interaction of drug intervention and time. In rhBNP group, MOU was significantly decreased with drug administraion (T2-T4 vs T1,mL O2/L: 10.61 ± 0.35,9.85 ± 0.60,9.79 ± 0.31 vs 11.59 ± 0.37). Conclusion Intravenous administration of rhBNP in AMI-HF model could decrease MOU and PCWP while increase the cardiac output.