OBJECTIVES: To investigate the active components and possible mechanisms of n-butanol fraction of Periploca forrestii Schltr. ethanol extract for treating Alzheimer's disease (AD). METHODS: The active components of n-butanol fraction of Periploca forrestii Schltr. ethanol extract were analyzed using UPLC-QE-MS technique. In a SD rat model of AD induced by treatment with AlCl₃ and D-gal, the therapeutic effects of low, moderate and high doses of the n-butanol fraction, saline, and donepezil hydrochloride were evaluated using ELISA, HE and Nissl staining, immunohistochemistry and Western blotting. The therapeutic mechanisms of the n-butanol fraction were explored using network pharmacology and molecular docking. RESULTS: Seventeen active components were identified from the n-butanol fraction of Periploca forrestii Schltr. ethanol extract, including phenylpropanoids, flavonoids, anthraquinones, triterpenoids, steroids, and volatile oils. In the rat models of AD, treatment with the n-butanol fraction significantly lowed AChE content in the hippocampus, increased the contents of ACh, SOD, CAT, and GSH-Px, enhanced the expressions of neuronal apoptotic factors Bcl-2, PI3K, Akt, p-PI3K, and p-Akt, and reduced the expressions of Bax and caspase-3 proteins. The treatment also dose-dependently up-regulated hippocampal expressions of Nrf-2, HO-1 and BDNF and down-regulated Keap-1, Aβ and Tau expressions. Bioinformatics analysis identified 14 key intersected targets (including TNF, AKT1 and ESR1) between the n-butanol fraction and AD. CONCLUSIONS The therapeutic effect of n-butanol fraction of Periploca forrestii Schltr. ethanol extract in AD mice is mediated by its multiple active components that regulate multiple targets and pathways.
Animals ; Rats, Sprague-Dawley ; Rats ; 1-Butanol/chemistry* ; Plant Extracts/pharmacology* ; Periploca/chemistry* ; Ethanol/chemistry* ; Alzheimer Disease/drug therapy* ; Male ; Molecular Docking Simulation ; Apoptosis/drug effects*

Spinal cord injury(SCI)can result in varying degrees of spinal motor dysfunction,partial sensory loss and sphincter dysfunction.SCI is divided into two stages:primary injury and secondary injury.The spinal cord cell necrosis during the primary injury period and apoptosis,oxidative stress and autophagy during the secondary injury period lead a large number of cell injury and permanant neurodysfunction.The histone deacetylase(HDAC)plays a key role in regulating the cellular viability and gene transcription.Neuro-dysfunction induced by SCI is associated with transcriptional dysfunction associated with unbalanced levels of protein acetylation.Valproic acid(VPA)is a inhibitor of HDAC and is usually used as an antiepileptic drug in clinic.Studies show that VPA may have the potential to treat the central nervous system diseases.VPA inhib-its HDAC,and then regulates oxidative stress,cellular autophagy,ion imbalance,microglia differentiation and inflammatory response,and plays the neuroprotective effect.This paper reviewed the related molecular mecha-nism of VPA in treating SCI.

Objective To explore the preventive effect and protective mechanism of electroacupuncture at Neiguan points on acute high altitude hypoxic brain injury by detecting the expression of EPO,p-PI3K,p-AKT and Bcl-2 proteins.Methods 48 male SD rats of SPF grade were randomly divided into control group,model group,western medicine group and Neiguan group.The acute high altitude hypoxic brain injury model was established using a hypobaric hypoxia chamber.The Western medication group received Citicoline Sodium via gastric gavage administration,while the Neiguan acupoint group was administered bilateral electroacupuncture stimulation at the Neiguan acupoints.The morphological changes of hippocampal tissues were observed by HE method,serum biochemical indexes were determined by ELISA method,neuronal apoptosis of rat hippocampal tissues was observed by TUNEL method and the positive rate was calculated,and the expression levels of EPO,p-PI3K,p-AKT and Bcl-2 proteins were detected in rat hippocampal tissues by Western-blot method.Results Compared with the control group,the model group exhibited extensive necrotic pyramidal cells with pyknotic and deeply stained nuclei,significantly elevated serum levels of GFAP,S100B,and UCH-L1(P<0.01),markedly increased hippocampal neuronal apoptosis(P<0.01),and upregulated protein levels of EPO,p-PI3K,p-AKT,and Bcl-2 in hippocampal tissue(P<0.01).Compared with the model group,both the western medication group and the electroacupuncture at Neiguan group exhibited a significant reduction in necrotic pyramidal cells in brain tissue,with occasional pyknotic and deeply stained nuclei.Serum levels of GFAP,S100B,and UCH-L1 were markedly decreased(P<0.01),hippocampal neuronal apoptosis was significantly reduced(P<0.01),and protein expression of EPO,p-PI3K,p-AKT,and Bcl-2 in hippocampal tissue was upregulated(P<0.01).No statistically significant differences were observed between the western medication group and the Neiguan electroacupuncture group(P>0.05).Conclusion Electroacupuncture at Neiguan may ameliorate acute high-altitude hypoxic brain injury,potentially through modulating EPO protein expression to activate the PI3K/AKT signaling pathway,thereby inhibiting hippocampal neuronal apoptosis.

Objective To investigate the effects of electroacupuncture at Neiguan(PC6)combined with Buyang Huanwu Decoction on myocardial edema-related proteins and its cardioprotective role in mice with acute high-altitude hypoxic myocardial injury,and to explore the potential mechanisms by which this combined therapy ameliorates acute hypoxic myocardial damage.Methods Mice were randomly divided into control,hypoxia model,electroacupuncture at Neiguan,Buyang Huanwu Decoction,and electroacupuncture at Neiguan+Buyang Huanwu Decoction groups.Except for the normal control group,all other groups were subjected to the establishment of an acute high-altitude hypoxia-induced myocardial injury model.Four days before entering the low-pressure hypoxia animal simulation chamber,the electroacupuncture at Neiguan group was treated with bilateral electroacupuncture at Neiguan,the Buyang Huanwu Decoction group was treated with Buyang Huanwu Decoction by gavage,and the electroacupuncture at Neiguan+Buyang Huanwu Decoction group was treated with a combination of electroacupuncture at Neiguan and Buyang Huanwu Decoction.The intervention lasted for 7 days.The normal control group and the hypoxia model group were handled normally without any other treatment.Myocardial pathology and ultrastructure were evaluated using HE staining and transmission electron microscopy.Serum levels of creatine kinase-MB(CK-MB)and cardiac troponin I(cTn-I)were measured by ELISA.Western blot was performed to quantify β1-AR,cAMP,PKA,and AQP1 protein expression.Results Compared with normal control group,the hypoxia model group exhibited significant myocardial damage,elevated cardiac biomarkers,and upregulated β1-AR/cAMP/PKA pathway proteins with increased AQP1 expression(all P<0.01).The electroacupuncture at Neiguan+Buyang Huanwu Decoction group demonstrated attenuated myocardial injury,reduced biomarker levels,and downregulated target proteins(all P<0.01)versus the hypoxia model group.Conclusion Electroacupuncture at Neiguan combined with Buyang Huanwu Decoction alleviates myocardial edema and injury in acute hypobaric hypoxia by reducing vascular permeability,potentially via suppression of the β1-AR/cAMP/PKA pathway and subsequent inhibition of AQP1 expression.

Objective This study aimed to identify PAX9 variants in non-syndromic tooth agenesis families of Chi-na,as well as to analyze the genotype-phenotype of non-syndromic tooth agenesis caused by PAX9 variants,which can provide a basis for the genetic diagnosis of tooth agenesis.Methods We collected the data of 44 patients with non-syn-dromic oligodontia who underwent treatment at Stomatological Hospital of Hebei Medical University between 2018 and 2023.Whole-exome sequencing was performed on the peripheral blood of the proband and its core family members,and the variants were verified by Sanger sequencing.Pathogenicity analysis and function prediction of the variants were per-formed using bioinformatics tools.The correlation between the genotype of PAX9 variant and its corresponding pheno-type was examined by reviewing 55 publications retrieved from PubMed.The studies involved 232 tooth agenesis pa-tients with PAX9 variants.Results A novel PAX9 c.447delG(p.Pro150Argfs*62)and a reported PAX9 c.406C>T(p.Gln136*)were identified in two Chinese families.Through bioinformatics analysis and three-dimensional structural mod-eling,we postulated that the frameshift variant was pathogenic.The outcome was the premature cessation of PAX9 pro-tein,which caused severe structural and functional deficiencies.Summarizing the PAX9 genotype-phenotype relationship revealed that patients carrying the PAX9 variant commonly led to loss of the second molars.Conclusion We identified the novel PAX9 c.447delG(p.Pro150Argfs*62)in a Chinese family of non-syndromic oligodontia,expanding the known variant spectrum of PAX9.The most susceptible tooth position for PAX9 variants of tooth agenesis was the second mo-lars and the deciduous molars during the deciduous dentition.

Objective:To automatically segment diabetic retinal exudation features from deep learning color fundus images.Methods:An applied study. The method of this study is based on the U-shaped network model of the Indian Diabetic Retinopathy Image Dataset (IDRID) dataset, introduces deep residual convolution into the encoding and decoding stages, which can effectively extract seepage depth features, solve overfitting and feature interference problems, and improve the model's feature expression ability and lightweight performance. In addition, by introducing an improved context extraction module, the model can capture a wider range of feature information, enhance the perception ability of retinal lesions, and perform excellently in capturing small details and blurred edges. Finally, the introduction of convolutional triple attention mechanism allows the model to automatically learn feature weights, focus on important features, and extract useful information from multiple scales. Accuracy, recall, Dice coefficient, accuracy and sensitivity were used to evaluate the ability of the model to detect and segment the automatic retinal exudation features of diabetic patients in color fundus images.Results:After applying this method, the accuracy, recall, dice coefficient, accuracy and sensitivity of the improved model on the IDRID dataset reached 81.56%, 99.54%, 69.32%, 65.36% and 78.33%, respectively. Compared with the original model, the accuracy and Dice index of the improved model are increased by 2.35%, 3.35% respectively.Conclusion:The segmentation method based on U-shaped network can automatically detect and segment the retinal exudation features of fundus images of diabetic patients, which is of great significance for assisting doctors to diagnose diseases more accurately.

Objective To investigate the therapeutic effect of traditional Chinese medicine (TCM) on portal vein thrombosis (PVT) in patients with liver cirrhosis and its medication characteristics. Methods A retrospective analysis was performed for 89 patients with liver cirrhosis and PVT who were hospitalized and treated in Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, and according to whether TCM treatment was applied in combination, they were divided into TCM group with 59 patients and control group with 30 patients. Related data were collected for the two groups, including demographic data, laboratory examination, radiological examination, gastroscopy, history of surgery, portal hypertension-related complications, medication, and follow-up data. The independent samples t -test was used for comparison of normally distributed continuous data between two groups, and the Mann-Whitney U rank sum test was used for comparison of non-normally distributed continuous data between two groups; the chi-square test or the Fisher's exact test was used for comparison of categorical data between two groups. An ordinal polytomous Logistic regression analysis was used for multivariate analysis. TCM Inheritance Computing Platform (V3.0) was used to perform a drug effect cluster analysis of TCM prescriptions. Results The multivariate logistic regression analysis showed that esophageal and gastric varices (odds ratio [ OR ]=3.144, 95% confidence interval [ CI ]: 1.221-8.094), PVT involving the portal vein (PV) and the superior mesenteric vein (SMV) ( OR =51.667, 95% CI : 3.536-754.859), PVT involving PV+spleen vein (SV)+SMV ( OR =13.271, 95% CI : 2.290-76.928), cavernous transformation of the portal vein ( OR =11.896, 95% CI : 1.172-120.696), and TCM intervention ( OR =0.348, 95% CI : 0.129-0.938) were influencing factors for the outcome of PVT in liver cirrhosis. Follow-up results showed that compared with the control group, the TCM group had a significantly lower progression rate (16.95% vs 56.67%, P < 0.001) and a significantly lower incidence rate of variceal rupture and bleeding (8.47% vs 33.33%, P < 0.001). Effective TCM drugs with a relatively high frequency of use included deficiency-tonifying drugs (359 times, 34.6%), blood-activating and stasis-resolving drugs (202 times, 19.5%), and diuresis-inducing and dampness-draining drugs (180 times, 17.3%); the TCM drugs with a relatively high frequency of use included Astragalus membranaceus (57 times, 8.7%), Angelica sinensis (50 times, 7.6%), and leech (48 times, 7.3%); TCM drug combinations with a relatively high frequency of use included Astragalus membranaceus+Angelica sinensis, Astragalus membranaceus+leech, Angelica sinensis+leech, and Astragalus membranaceus+Angelica sinensis+leech. Conclusion Qi-tonifying, blood-activating, and stasis-breaking drugs, such as Astragalus membranaceus, Angelica sinensis, and leech, can promote the stabilization or recanalization of PVT in liver cirrhosis and reduce the incidence rate of bleeding events due to portal hypertension.

OBJECTIVE To establish comprehensive quality evaluation method based on multi-index components combined with multivariate statistical analysis， and to comprehensively evaluate the quality of Periploca forrestii. METHODS Taking 11 batches of P. forrestii medicinal materials from different areas in Guizhou as samples， the contents of neochlorogenic acid， cryptochlorogenic acid， chlorogenic acid， procyanidin A2， isochlorogenic acid A and isochlorogenic acid C were determined by HPLC. Clustering heat map analysis， grey correlation analysis（GRA） and technique for order preference by similarity to ideal solution（TOPSIS） were used to evaluate the quality of P. forrestii. RESULTS The results of methodological investigation of content determination were in accordance with the relevant regulations， and the linear relationship and accuracy of each component were good in their respective sampling range. The contents of chlorogenic acid， cryptochlorogenic acid， neochlorogenic acid， procyanidin A2， isochlorogenic acid A and isochlorogenic acid C in 11 batches of samples were 3.650-7.302， 0.888-2.575， 1.371- 2.386， 0.947-1.469， 0.084-0.169 and 0.725-1.067 mg/g， respectively. The content of each component was significantly different， with the highest content of chlorogenic acid and the lowest content of isochlorogenic acid A. The comprehensive results of cluster heat map， GRA and TOPSIS analysis showed that the comprehensive quality of S5 and S10 was relatively good. CONCLUSIONS The established method is accurate， stable and simple. Combined with multivariate statistical analysis method， it can be used for quality evaluation of P. forrestii. The quality of samples from Jiuzhou Town and Caiguan Town of Xixiu District in Anshun City of Guizhou Province are relatively good among 11 different origin samples.

OBJECTIVE To provide reference for quality control of Gentiana rhodantha. METHODS Taking 52 batches of G. rhodantha as subject， ultra-high performance liquid chromatography （UPLC） fingerprint was adopted. The similarity of 52 batches of medicinal materials samples was evaluated by the Similarity Evaluation System for Chromatographic Fingerprints of Traditional Chinese Medicine （2004A edition）； the content of mangiferin was determined； chemometric analyses [cluster analysis， principal component analysis （PCA） and orthogonal partial least squares-discriminant analysis （OPLS-DA）] were performed. RESULTS UPLC fingerprints of 52 batches of G. rhodantha were established， 17 common peaks were identified， and 6 of them were identified， which were loganic acid （peak 1）， neomangiferin （peak 3）， swertiamarin （peak 5）， dangyin （peak 6）， mangiferin （peak 7） and isoorientin （peak 9）. The similarities of 52 batches of medicinal materials samples were all greater than 0.9； cluster analysis showed that S1-S46， S48-S52 clustered into one class， and S47 alone； PCA results showed that the cumulative variance contribution rate of the first six principal components was 82.928%； OPLS-DA results showed that the corresponding components of swertiamarin， mangiferin and chemical composition represented by peak 4， 14， 15， 16 were the main iconic components affecting the quality differences of G. rhodantha medicinal materials. The contents of mangiferin in 52 batches of medicinal material samples ranged from 18.2 to 101.0 mg/g， mostly in accordance with 2020 edition of Chinese Pharmacopoeia. CONCLUSIONS The established UPLC fingerprint and chemometric analysis methods combined with content determination method of mangiferin can comprehensively evaluate the quality of G. rhodantha.

OBJECTIVE To prepare celastrol -loaded albumin nanoparticles （CLT-AN），and to investigate their activity against rheumatoid arthritis （RA）in vivo . METHODS CLT-AN was prepared by ultrasonic method . The formulation technology was optimized by single -factor test by taking particle size ，polydispersity index （PDI）and stability as indexes ，with the dosage of CLT ， the dosage of soybean oil and the ultrasonic power as factors . The physical and chemical properties of CLT -AN were investigated by transmission electron microscopy （TEM）and laser particle size analyzer ；in vitro stability and release profile were studied . A rat model of adjuvant -induced arthritis was constructed to investigate the effects of CLT -AN on joint swelling ，the levels of serum inflammatory factors [tumor necrosis factor α（TNF-α）and interleukin -1β（IL-1β）] and pathological state of joint tissue . RESULTS The optimized formulation was CLT 6.5 g，soybean oil 45 mg，ultrasonic power 490 W，ultrasonic time 8 min. CLT- AN prepared by the best formulation showed uniform and spherical morphology . Its particle size ，PDI，Zeta potential were （96.8± 1.1）nm，0.174±0.020，and（－18.6±1.7）mV，respectively. The encapsulation efficiency and drug -loading efficiency were （94.61±0.46）% and（2.42±0.21）%. There were no significant changes in particle size ，PDI，Zeta potential and encapsulation efficiency of CLT -AN within 5 days of storage at room temperature . CLT-AN was slowly released in vitro ，and the cumulative release reached 73.56% in 72 h. Compared with CLT ，CLT-AN could significantly inhibit the joint swelling of model rats ，reduced the levels of inflammatory factors TNF -α and IL -1β in serum ，and improved the pathological state of inflammatory joint tissue . CONCLUSIONS CLT-AN prepared by ultrasonic method has the appropriate particle size ，good stability ，significant sustained - release characteristics ，and excellent therapeutic efficacy against RA .