Objective To evaluate the effect of Kaixin San(KXS)combined with fluoxetine on hippocampal neural stem cells in mice with chronic stress stress and depression.Methods A mouse model of depression was constructed using the method of chronic unpredictable stress stress,and the highest dose of KXS water extract and fluoxetine for clinical application was given for 28 days,and behavioral tests were carried out.Nissl staining was used to detect the pathological status of hippocampal tissues in mice.The expression of TUNEL and Nestin in mouse hippocampus was determined by immunofluorescence.Western blotting was used to detect the expressions of apoptotic proteins cleaved caspase-3 and caspase-3,pyroptosis proteins GSDMD and cleaved caspase-1,as well as the expression of neural stem cell marker Nestin in the hippocampus,and the expression of Wnt/β-catenin signaling pathway-related proteins in the hippocampus.Results The combination of KXS extract and fluoxetine can significantly improve the depression-like behavior of model mice,and the effect is better than fluoxetine alone.The combination inhibited the activation of apoptosis and pyroptosis signaling pathways in the hippocampus when used alone with high-dose fluoxetine,significantly upregulated the expression of Nestin,and regulated the expression of Wnt/β-catenin signaling pathway protein.Conclusion The combination of KXS and high-dose fluoxetine can improve apoptosis and pyroptosis in the hippocampus of stress stress and depression model mice,and upregulate the expression of neural stem cell marker Nestin by regulating the Wnt/β-catenin signaling pathway,which may be a key link to improve the antidepressant effect of the combination drug.

Objective To explore the relationship between the severity of white matter hyperintensities(WMH)and the distribution of collateral disease syndrome types.Methods A total of 208 patients clearly diagnosed with cerebral small vessel disease in Lishui TCM Hospital Affiliated to Zhejiang Chinese Medical University from December 2022 to December 2023 were selected as the study subjects.All enrolled subjects in the study completed the collateral disease syndrome differentiation.WMH was graded on magnetic resonance imaging fluid-attenuated inversion recovery(FLAIR)sequences using the Fazekas scale and measured for volume.Results Among the 208 patients,there were 52 cases of the brain collateral impoverishment(BCI)type,with Fazekas score of 3(2,3)points,and WMH volume of(8.80±9.37)ml;67 cases of the brain collateral stasis(BCS)type,with Fazekas score of 4(3,4)points,and WMH volume of(13.42±9.18)ml;47 cases of the brain collateral constraint(BCC)type,with Fazekas score of 4(3,5)points,and WMH volume of(14.60±10.07)ml;and 42 cases of the brain collateral occlusion(BCO)type,with Fazekas score of 4(4,5)points,and WMH volume of(18.65±10.71)ml.Of these,55 patients with a Fazekas score of 1-2 points were most commonly of the BCI type,105 patients with a Fazekas score of 3-4 points were most commonly of the BCS type,and 48 patients with a Fazekas score of 5-6 points were most commonly of the BCO type.There were statistically significant differences in Fazekas scores and WMH volumes between the BCI and BCO types and other syndrome types(P＜0.05).Conclusion The severity of WMH corresponds to a certain distribution of collateral disease syndrome types,and can serve as a reference indicator for assessing the severity of illness in patients with collateral disease.

Objective To evaluate the effect of Kaixin San(KXS)combined with fluoxetine on hippocampal neural stem cells in mice with chronic stress stress and depression.Methods A mouse model of depression was constructed using the method of chronic unpredictable stress stress,and the highest dose of KXS water extract and fluoxetine for clinical application was given for 28 days,and behavioral tests were carried out.Nissl staining was used to detect the pathological status of hippocampal tissues in mice.The expression of TUNEL and Nestin in mouse hippocampus was determined by immunofluorescence.Western blotting was used to detect the expressions of apoptotic proteins cleaved caspase-3 and caspase-3,pyroptosis proteins GSDMD and cleaved caspase-1,as well as the expression of neural stem cell marker Nestin in the hippocampus,and the expression of Wnt/β-catenin signaling pathway-related proteins in the hippocampus.Results The combination of KXS extract and fluoxetine can significantly improve the depression-like behavior of model mice,and the effect is better than fluoxetine alone.The combination inhibited the activation of apoptosis and pyroptosis signaling pathways in the hippocampus when used alone with high-dose fluoxetine,significantly upregulated the expression of Nestin,and regulated the expression of Wnt/β-catenin signaling pathway protein.Conclusion The combination of KXS and high-dose fluoxetine can improve apoptosis and pyroptosis in the hippocampus of stress stress and depression model mice,and upregulate the expression of neural stem cell marker Nestin by regulating the Wnt/β-catenin signaling pathway,which may be a key link to improve the antidepressant effect of the combination drug.

Objective To explore the relationship between the severity of white matter hyperintensities(WMH)and the distribution of collateral disease syndrome types.Methods A total of 208 patients clearly diagnosed with cerebral small vessel disease in Lishui TCM Hospital Affiliated to Zhejiang Chinese Medical University from December 2022 to December 2023 were selected as the study subjects.All enrolled subjects in the study completed the collateral disease syndrome differentiation.WMH was graded on magnetic resonance imaging fluid-attenuated inversion recovery(FLAIR)sequences using the Fazekas scale and measured for volume.Results Among the 208 patients,there were 52 cases of the brain collateral impoverishment(BCI)type,with Fazekas score of 3(2,3)points,and WMH volume of(8.80±9.37)ml;67 cases of the brain collateral stasis(BCS)type,with Fazekas score of 4(3,4)points,and WMH volume of(13.42±9.18)ml;47 cases of the brain collateral constraint(BCC)type,with Fazekas score of 4(3,5)points,and WMH volume of(14.60±10.07)ml;and 42 cases of the brain collateral occlusion(BCO)type,with Fazekas score of 4(4,5)points,and WMH volume of(18.65±10.71)ml.Of these,55 patients with a Fazekas score of 1-2 points were most commonly of the BCI type,105 patients with a Fazekas score of 3-4 points were most commonly of the BCS type,and 48 patients with a Fazekas score of 5-6 points were most commonly of the BCO type.There were statistically significant differences in Fazekas scores and WMH volumes between the BCI and BCO types and other syndrome types(P＜0.05).Conclusion The severity of WMH corresponds to a certain distribution of collateral disease syndrome types,and can serve as a reference indicator for assessing the severity of illness in patients with collateral disease.

Objective To investigate the beneficial effect of Kai-Xin-San combined with fluoxetine in improving depression-like behaviors on chronic unpredictable mild stress(CUMS)induced depression model mice.Methods The present study aimed to assess the potential of Kai-Xin-San in combination with fluoxetine to ameliorate depression-like behaviors in a CUMS induced mouse depression model.Behavioral tests,such as the sucrose preference test were employed to evaluate the efficacy of the treatment.Additionally,the levels of suppressed stress factors were measured using the ELISA method.The morphology of hippocampal tissue was evaluated using the HE staining method,Nissl Staining and TUNEL staining methods.Furthermore,western blotting analysis was utilized to determine the expression levels of proteins such as Caspase-3,and Caspase-9.Results The co-administration of Kai-Xin-San and fluoxetine resulted in a significant increase in sucrose preference rate in model mice.This effect was comparable to that of fluoxetine alone at the standard clinical dose.Furthermore,the combination treatment up-regulated the levels of suppressed stress factors,reduced the apoptosis of hippocampus induced by depression and regulated the apoptosis signaling pathway in hippocampus.Conclusion The combination of Kai-Xin-San and fluoxetine has been shown to be an effective treatment for depression-like behavior in animal models,resulting in a reduction in the required clinical dosage of fluoxetine.This effect may be attributed to the up-regulation of neurotransmitter expression,inhibition of stress axis activation,and central nervous inflammation.

Objective To establish a method for quality evaluation of Nardostachyos Radix et Rhizoma derived from Nardostachys jatamansi DC by determining the contents of nardosinone and establishing fingerprints. Methods HPLC fingerprint and content determination methods were established to evaluate ten batches of Nardostachyos Radix et Rhizoma from Nardostachys jatamansi DC.For the determination of nardosinone, the samples were separated by Phenomenex-C18 ( 4. 6 mm × 250 mm, 5 μm) with the mobile phase consisting of acetonitrile and water at the flow rate of 0. 8 mL · min-1 , the detection wavelength was set at 250 nm,and column temperature was 25℃.The gradient mobile phase system was used for the fingerprints. Results HPLC fingerprint of Nardostachyos Radix et Rhizoma was established with good chromatography separation and repeatability, which could be used for quality control of Nardostachyos Radix et Rhizoma. After similarity analysis, the results showed that there was no significantly difference between the HPLC fingerprints of samples from different origins,but the content of four major peaks were different.The amount of nardosinone from ten batches of Nardostachyos Radix et Rhizoma was determined by HPLC-DAD, which showed that the contents of nardosinone from Nardostachys jatamansi DC were obviously different with each other. Conclusion The method is sensitive, repeatable and accurate. It can be used as a method of quality control for Nardostachyos Radix et Rhizoma.

OBJECTIVE:To conduct screening for the active ingredients of Ziziphus jujuba with neuroprotective effect and to il-luminate their mechanisms of action preliminarily. METHODS:After neuron-like cells (PC12 cells) were respectively cultured in the ingredient of Z. jujuba with polysaccharide enriched(1 mg/ml),that with polysaccharide removed(1 mg/ml),7 kinds of flavo-noid ingredients of Z. jujuba(catechin,procyanidine B2,epicatechin,hyperoside,rutin,quercetin-3-β-D-glucoside and kaempfer-ol-3-O-rutinoside,represented by A,B,C,D,E,F,G,all at the concentrations of 3,13,30 μmol/L)and 2 kinds of nucleoside ingredients of Z. jujuba (cyclic adenosine monophosphate and cyclic guanosine monophosphate,both at the concentrations of 3, 13,30 μmol/L)for 24 h,tert-butyl hydroperoxide(tBHP,150 μmol/L)was used to act on PC12 cells for 3 h to induce oxidative cellular damage,and MTT assay was employed to detect the survival rate of PC12 cells. The PC12 cells transfected with reporter gene plasmid(pARE-Luc)were cultured as above for 24 h,luciferase(Luc)assay was used to detect the transcriptional levels of the antioxidant response element(ARE)of all groups of cells(reflected as the activity of Luc)so as to investigate the anti-injury mechanism. RESULTS:The ingredient of Z. jujuba with polysaccharide enriched could significantly increase the survival rate of PC12 cells to which oxidative damage was caused and the transcriptional level of ARE in the transfected cells. Among the flavonoid ingredients of Z. jujuba, A(30 μmol/L),B(3-30 μmol/L)and C(10-30 μmol/L)could significantly increased the survival rate of the cells,and A(30 μmol/L),B(3-30 μmol/L),C(30 μmol/L),E(30 μmol/L)and F(3-30 μmol/L)could obviously in-creased the activation level of ARE in the transfected cells. However,the nucleoside ingredients of Z. jujuba including cyclic ade-nosine monophosphate and cyclic guanosine monophosphate had no obvious effect of increasing the survival rate of PC12 cells to which oxidative damage was caused or activating the transcription of ARE in the transfected cells. CONCLUSIONS:The polysac-charide and flavonoid ingredients of Z. jujuba may be the active ingredients which account for the neuroprotective effect against oxi-dative cellular damage,and their mechanisms of action may be related to the activation of ARE transcription.

Objective To provide scientific evidence for the identification and development of fructus psoraleae through its pharmacognostical study.Methods To identify fructus psoraleae by paraffin section and powder method,meanwhile,morphological identification and physical and chemical identification were also adopted.Results The morphological and microscopic characteristics of transverse section and powder of fructus psoraleae were described; fluorescent speckles of the same color were found in the same place of test samples and psoralen and isopsoralen reference by thin layer chromatography; results of water and ash determination all complied with Chinese pharmacopoeia specification,water and 70％ ethanol extractives were 21.93～39.68％,22.03～31.77％,respectively.Conclusion The established method was simple and practical with reliable results,and suitable for the identification of fructus psoraleae.

Objective To investigate the influence of super fine crushing technique on dissolution rate of Sunshang Capsule (SC). Methods With the dissolution of active components and notoginsengside as the markers, the dissolution rate in Sunshang Capsule and Sunshang powder which were prepared by super fine crushing method and fine crushing method respectively was compared by TLC and HPLC. Results The stain of Sunshang Capsule was bigger and darker than that of Sunshang Powder, time-limited dissolution of SC was 4.66 %higher and the content of notoginsengside R1 was 6 %higher than those of Sunshang Powder. Conclusion Super fine crushing technique can increase the dissolution rate of SC and this will supply evidence for the process of SC preparation.

98 %).Conclusions Saponins are the major active ingredients in Radix Polygalae.Tenuifolin as an alkaline hydrolysis product of total saponins can be used as a chemical reference substance for the quality control of Radix Polygalae.