1.Research progresses in gene therapy for hepatolenticular degeneration
Gui CHEN ; Haoyang ZHENG ; Fang LIU ; Jing YUAN ; Yuanhong XU ; Weisheng CHENG
Chinese Journal of Hepatology 2025;33(1):97-102
Hepatolenticular degeneration, also known as Wilson's disease, is a type of autosomal recessive genetic disorder of copper metabolism. The causative gene, ATP7B, is located on the long arm of chromosome 13 and encodes a P-type ATPase that is involved in copper transport. Pathogenic mutations in the ATP7B gene sequence lead to the diminished or lost function of the ATP7B protein, resulting in pathological copper deposition in organs such as the liver, brain, kidneys, and cornea. Currently, the treatment of Wilson's disease primarily involves oral medications to promote copper excretion or reduce copper absorption so as to alleviate the state of illness. However, pharmacological treatment has objective limitations, including the need for lifelong therapy and varying degrees of adverse drug reactions in some patients. Gene therapy can fully correct the genetic defect, restore ATP7B protein function, achieve a curative effect, and improve the patient's quality of life.
2.Study on the Spectral Effect Relationship of Antioxidant Activity of Mori Folium
Mengwen LIU ; Qiaoling DENG ; Jing SHEN ; Xuanshi CHEN ; Hui XIAO ; Weisheng XU
Herald of Medicine 2025;44(3):387-391
Objective To determine the material basis of the antioxidant activity of Mori Folium by examining the spec-trum-effect relationship.Methods High-performance liquid chromatography(HPLC)was utilized to establish the fingerprints of Mori Folium.The antioxidant activity of Mori Folium was assessed using the 1,1-diphenyl-2-picrylhydrazyl(DPPH)radical scavenging assay and other related indicators.The spectrum-effect relationship of antioxidation was analyzed using gray relational analysis,bivariate correlation analysis,and partial least squares regression analysis.Molecular docking techniques were employed to predict potential interaction targets.Results HPLC fingerprints for 13 batches of Mori Folium were established,and thirteen common peaks were marked,with similarities ranging from 0.932 to 0.998.Nine common peaks were identified by comparing them to reference substances.Differences in antioxidant activity were observed among the different batches of Mori Folium.Based on the analysis of the spectrum-effect relationship,chemical components such as chlorogenic acid,cryptochlorogenic acid,rutin,and iso-chlorogenic acid B were found to contribute significantly to the antioxidant activity.These components may exert their effects by binding to several antioxidant protein targets,such as XOD,NO-1,and PPAR-α.This implies that Mori Folium might exert its an-tioxidant action via multiple components and targets.Conclusions By integrating the fingerprint and antioxidant activity of Mori Folium,the contributions of individual components to its antioxidant activity were determined.This study provides an experi-mental basis for elucidating the substances responsible for the antioxidant activity of Mori Folium and for establishing quality con-trol methods.
3.Exploration on the material basis and mechanism of Prunus mume f. viridicalyx for anti-depression based on UPLC-QE-Orbitrap-MS combined with network pharmacology
Weisheng LYU ; Cuijie WEI ; Yueyi LIANG ; Tianrui XIA ; Dongmei SUN ; Xiangdong CHEN ; Xiaozhou JIA
International Journal of Traditional Chinese Medicine 2025;47(6):822-832
Objective:To identify the components of Prunus mume f. viridicalyx based on ultra performance liquid chromatography-QE-Orbitrap mass spectrometry (UPLC-QE-Orbitrap-MS); To predict and analyze its substances and mechanisms to exert anti-depression effects combined with network pharmacology.Methods:UPLC-QE Orbitrap MS technology was used to analyze the chemical components of Prunus mume f. viridicalyx. Based on ChemSpider, mzCloud online platform, orbitrap TCM library and existing literature research, the secondary mass spectra of target compounds were compared and confirmed to identify the chemical composition of Prunus mume f. viridicalyx. The active components of the Prunus mume f. viridicalyx were screened. The Swiss Target Prediction database was used to predict targets with high correlation to active components in Prunus mume f. viridicalyx, and obtaining depression related disease targets from GeneCards and DisGeNET databases. The intersection targets of constituents and diseases were obtained using Venny platform. Protein-protein interaction network (PPI) was constructed by using String database, and the core targets were screened. Gene ontology function and Kyoto encyclopedia of genes and genomes pathway enrichment analysis of potential core targets were performed by using David database, and "active component-core target-signal pathway" network was constructed. PyMOL software was used to perform molecular docking between active components and key targets.Results:A total of 54 components, including organic acids, flavonoids and their glycosides, alkaloid, amino acids and other compounds were identified from Prunus mume f. viridicalyx. A total of 22 active components were screened and 92 active components and disease intersection targets were identified. A total of 13 core targets were screened through PPI network, including tumor necrosis factor, albumin, amyloid beta-protein precursor, AKT serine/threonine kinase 1 and so on. Enrichment analysis showed that Prunus mume f. viridicalyx mainly participated in transcription from RNA polymerase Ⅱ promoter, gene expression, protein binding and other functions, and presented the effects of anti-depression through MAPK, Toll-like receptor signaling pathway and other pathways. 12 key targets and 7 key active components were further obtained through the analysis of the "active component-core target-signal pathway" network, three of them were confirmed as kaempferol, quercetin, and isorhamnetin by reference substance. Molecular docking showed that 3 compounds could bind to the target proteins of depression well.Conclusion:Prunus mume f. viridicalyx exerts antidepressant effects through multiple components, targets, and pathways, mainly through the MAPK signaling pathway.
4.Study on the Spectral Effect Relationship of Antioxidant Activity of Mori Folium
Mengwen LIU ; Qiaoling DENG ; Jing SHEN ; Xuanshi CHEN ; Hui XIAO ; Weisheng XU
Herald of Medicine 2025;44(3):387-391
Objective To determine the material basis of the antioxidant activity of Mori Folium by examining the spec-trum-effect relationship.Methods High-performance liquid chromatography(HPLC)was utilized to establish the fingerprints of Mori Folium.The antioxidant activity of Mori Folium was assessed using the 1,1-diphenyl-2-picrylhydrazyl(DPPH)radical scavenging assay and other related indicators.The spectrum-effect relationship of antioxidation was analyzed using gray relational analysis,bivariate correlation analysis,and partial least squares regression analysis.Molecular docking techniques were employed to predict potential interaction targets.Results HPLC fingerprints for 13 batches of Mori Folium were established,and thirteen common peaks were marked,with similarities ranging from 0.932 to 0.998.Nine common peaks were identified by comparing them to reference substances.Differences in antioxidant activity were observed among the different batches of Mori Folium.Based on the analysis of the spectrum-effect relationship,chemical components such as chlorogenic acid,cryptochlorogenic acid,rutin,and iso-chlorogenic acid B were found to contribute significantly to the antioxidant activity.These components may exert their effects by binding to several antioxidant protein targets,such as XOD,NO-1,and PPAR-α.This implies that Mori Folium might exert its an-tioxidant action via multiple components and targets.Conclusions By integrating the fingerprint and antioxidant activity of Mori Folium,the contributions of individual components to its antioxidant activity were determined.This study provides an experi-mental basis for elucidating the substances responsible for the antioxidant activity of Mori Folium and for establishing quality con-trol methods.
5.Research progresses in gene therapy for hepatolenticular degeneration
Gui CHEN ; Haoyang ZHENG ; Fang LIU ; Jing YUAN ; Yuanhong XU ; Weisheng CHENG
Chinese Journal of Hepatology 2025;33(1):97-102
Hepatolenticular degeneration, also known as Wilson's disease, is a type of autosomal recessive genetic disorder of copper metabolism. The causative gene, ATP7B, is located on the long arm of chromosome 13 and encodes a P-type ATPase that is involved in copper transport. Pathogenic mutations in the ATP7B gene sequence lead to the diminished or lost function of the ATP7B protein, resulting in pathological copper deposition in organs such as the liver, brain, kidneys, and cornea. Currently, the treatment of Wilson's disease primarily involves oral medications to promote copper excretion or reduce copper absorption so as to alleviate the state of illness. However, pharmacological treatment has objective limitations, including the need for lifelong therapy and varying degrees of adverse drug reactions in some patients. Gene therapy can fully correct the genetic defect, restore ATP7B protein function, achieve a curative effect, and improve the patient's quality of life.
6.Effect of deliberate practice combined with intensive training in cardiopulmonary resuscitation teaching for nurses receiving standardized training
Weiping WANG ; Qiuying DENG ; Chuanren LING ; Weisheng CHEN ; Haihui FANG ; Liqun HUANG
Chinese Journal of Medical Education Research 2024;23(1):123-127
Objective:To investigate the mid- and long-term training effect of deliberate practice combined with intensive training in cardiopulmonary resuscitation (CPR) teaching for nurses receiving standardized training.Methods:A total of 160 nurses who received the first year of standardized training in Guangdong Provincial Hospital of Traditional Chinese Medicine from July 2020 to March 2021 were enrolled as subjects, and they were randomly divided into experimental group and control group, with 80 nurses in each group. The nurses in the control group received the mode of single deliberate practice, and those in the experimental group received deliberate practice combined with intensive training at months 0, 3, and 6. Assessment was performed for the two groups before and after training,with objective indicators and subjective evaluation as the assessment criteria. Objective score was determined by the immediate objective feedback provided by the electronic display of Resusci Anne ? QCPR, and subjective score was determined by the same group of examiners based on the evaluation of cardiopulmonary resuscitation performed by trainees,including environment evaluation, judgment of consciousness, judgment of respiration, initiation of emergency treatment system, re-evaluation after resuscitation, and overall evaluation. SPSS 18.0 was used for the independent samples t-test, the non-parametric test, and the chi-squares test. Results:After training, there were significant differences between the experimental group and the control group in the objective indicators of the percentage of correct compression (CC%) [85.500 (77.50, 93.00) vs. 81.00 (71.75, 89.00), Z=-2.49, P=0.013] and the proportion of correct ventilation (V%) [84.00 (75.00, 92.75) vs. 80.00 (64.00, 90.25), Z=-2.15, P=0.031]. Both groups had significant improvements in objective and subjective indicators after training (all P<0.05). Conclusions:Deliberate practice combined with intensive training at an interval of 3 months can effectively improve the mid- and long-term CPR skills of nurses receiving standardized training and is more suitable for high-level muscle memory training.
7.Rehmanniae Radix Praeparata Improves Neurological Function of Ischemic Stroke Rats by Inhibiting Autophagy and Ferroptosis
Saifei LI ; Peipei YUAN ; Yaxin WEI ; Liyuan GAO ; Panying LI ; Yuan RUAN ; Yi CHEN ; Yang FU ; Xiaoke ZHENG ; Weisheng FENG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(8):26-33
ObjectiveTo investigate the effect of Rehmanniae Radix Praeparata on neurological function injury in ischemic stroke rats and explore its mechanism. MethodMale SD rats were randomized into sham operation, model, low- and high -dose (3.5 g·kg-1 and 7 g·kg-1) Rehmannia Radix Praeparata, and nimodipine (0.01 g·kg-1) groups. The rat model of middle cerebral artery occlusion (MCAO) was established with the modified suture occlusion method. Zea-Longa 5-point scoring was employed to evaluate the neurological function of rats. The cerebral infarction volume was detected by 2,3,5-triphenyltetrazolium chloride (TTC) staining. Hematoxylin-eosin staining and Nissl staining were employed to observe the morphology and damage of the brain tissue. Meanwhile, the serum levels of lactate dehydrogenase (LDH), oxidative stress-related indicators superoxide dismutase (SOD), glutathione peroxidase 4 (GPX4), and malondialdehyde (MDA), and the iron (Fe) content in the brain tissue were determined. To explore the mechanism of Rehmanniae Radix Preparata in mitigating the neurological damage in ischemic stroke rats, Western blotting was employed to determine the expression levels of proteins in the ischemic brain tissue. The autophagy-associated proteins included autophagy effector (beclin-1), microtubule-associated protein light chain 3 (LC3B), and ubiquitin-binding protein p62 (p62). The ferroptosis-associated proteins included transferrin (TF), transferrin receptor 1 (TFR1), ferritin heavy chain 1 (FTH1), and ferropotin (FPN1). The neurological function injury-associated proteins included brain-derived neurotrophic factor (BDNF) and tyrosine kinase receptor B (TrkB). ResultCompared with the sham operation group, the model group showed increased neurological function score, cerebral infarction volume, and appearance of nuclear pyknosis and vacuole of cells in the cerebral cortex. In addition, the model group presented elevated levels of LDH, MDA, and Fe (P<0.01) and lowered levels of SOD and GPX4 (P<0.01). Compared with the model group, Rehmanniae Radix Praeparata decreased the content of LDH, MDA, and Fe (P<0.05, P<0.01) and elevated the levels of SOD and GPX4 (P<0.05, P<0.01). Compared with the sham operation group, the modeling promoted the expression of beclin-1,LC3B Ⅱ/Ⅰ, TF, and TFR1 and inhibited the expression of p62, FTH1, FPN1, BDNF, and TrkB (P<0.01). The expression levels of these proteins were recovered after the treatment with Rehmanniae Radix Praeparata. ConclusionRehmanniae Radix Praeparata may inhibit ferroptosis and improve the neurological function in ischemic stroke rats by down-regulating the autophagy level in the brain tissue.
8.Establishment of UPLC characteristic chromatogram of Pulsatilla chinensis and its application in origin differentiation and counterfeit identification
Guangming HE ; Rui LUO ; Heping ZENG ; Xiaoying LU ; Xiaolong YANG ; Weisheng LYU ; Yueyi LIANG ; Zhenyu LI ; Dongmei SUN ; Xiangdong CHEN
International Journal of Traditional Chinese Medicine 2024;46(6):743-749
Objective:To establish ultra performance liquid chromatography (UPLC) characteristic chromatogram of Pulsatilla chinensis; To provide reference for the origin identification and quality control of Pulsatilla chinensis. Methods:UPLC Method was adopted. The determination was performed on a column of Agilent SB C18 (2.1 mm×100 mm, 1.8 μm) . The mobile phase was acetonitrile-methanol (2:1) -0.1% phosphoric acid solution by fradient elution at a flow rate of 0.30ml/min. The column temperature was 30 ℃. The detection wavelength was 215 nm. The injection volume was 2 μl. The common counterfeit products and medicinal herbs of Pulsatilla chinensis from different areas were evaluated by comparison of characteristic chromatogram, principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA). Results:There were 9 characteristic peaks in the characteristic chromatogram of Pulsatilla chinensis, and 8 common peaks were identified by high resolution mass spectrometry and comparison of reference materials. Through PCA analysis, it was possible to clearly distinguish the medicinal herbs of Pulsatilla chinensis from different areas. Combined with OPLS-DA analysis, it was found that peak 2, peak 3, peak 6 were the main markers of Pulsatilla chinensis from different producing areas. Conclusion:The established method has good specificity, repeatability and durability, and it can effectively distinguish the common counterfeits of Pulsatilla chinensis, and provide the basis of quality control and selection of origin for Pulsatilla chinensis.
9.Study on UPLC fingerprint of Mume flos at different flowering stages based on chemometrics analysis
Shuang HUANG ; Yueyi LIANG ; Jie YANG ; Weisheng LYU ; Xiaoying LU ; Guangming HE ; Zhipeng CHEN ; Xuxuan HOU ; Tianrui XIA ; Zhenyu LI ; Congyou DENG ; Xiangdong CHEN ; Dongmei SUN
International Journal of Traditional Chinese Medicine 2024;46(7):898-904
Objective:To establish the ultra high performance liquid chromatography (UPLC) fingerprints of Mume flos at different flowering stages; To provide reference for the quality research of Mume flos.Methods:The fingerprints of Mume flos were established by UPLC method, and the common peaks were identified by high performance liquid chromatography high resolution mass spectrometry (LC-MS). Chemometrics analysis was carried out with the fingerprints' common peak area of plum blossom at different flowering stages as a variable. Semiquantitative analysis of changes in flavonoids and phenolic acids in Mume flos at different flowering stages was conduct using peak area calculation method.Results:Totally 31 common peaks were identified in the fingerprints of plum blossom medicinal materials at different flowering stages and 9 components were identified. Clustering analysis (HCA) and principal component analysis (PCA) both classified plum blossom medicinal herbs at different flowering stages into three categories. Among them, there were significant differences between the groups at the bud stage, blooming period, and final flowering period, while the differences between the groups at blooming period and final flowering period were relatively small. The orthogonal partial least squares discriminant analysis (OPLS-DA) screened 16 different components with VIP>1.0. The contents of phenolic acids in different flowering stages were as follows: bud stage>blooming period>final flowering period, while the contents of flavonoids were as follows: blooming period>final flowering period>bud stage.Conclusions:This method is simple and reliable, and can provide reference for the quality evaluation of plum blossom medicinal materials at different flowering stages.
10.Screening and genotyping of Mur blood group among voluntary blood donors in the population of Hezhou,Guangxi
Weiquan YUAN ; Shaohua DING ; Jianmin LI ; Xueming WU ; Shengming WEN ; Houquan LIN ; Weisheng HE ; Xi-Aoming LI ; Jiajie ZHANG ; Longming XIAO ; Shengbao DUAN ; Shengwang CHEN
Chinese Journal of Blood Transfusion 2024;37(7):773-778
Objective To screen the distribution frequency of Mur blood group among voluntary blood donors in Hezhou,Guangxi,and further analyze the molecular basis of of Mur antigen positive samples.Methods The Mur pheno-type of voluntary blood donors in Hezhou was serologically screened using microplate method,and the distribution frequency of Mur antigens in different ethnic groups was analyzed.Genetic typing was performed on these positive samples with PCR-SSP method to verify the accuracy of the serological method,and the genetic background was sequenced and analyzed.Re-sults Among 3 298 samples from voluntary blood donors in Hezhou,432(13.10%,432/3 298)were screened positive for Mur antigen,and PCR-SSP genotyping validation showed that all 432 samples were electrophoretic positive.Among them,the proportion of Han blood donors with positive Mur antigen was12.79%(331/2 587),Yao ethnic group was13.25%(64/483),Zhuang ethnic group was 16.51%(36/218),and no statistically significant difference was found in the three groups(P>0.05).Further sequencing results showed that 428 samples were GYP(B-A-B)Mur,also known as GYP.Mur type(12.98%,428/3 298),the other 4 samples were GYP(B-A-B)Bun,also known as GYP.Bun type(0.12%,4/3 298).Conclusion The Mur blood type frequency is high in the voluntary blood donors in Hezhou,Guangxi,and is predominant characterized by GYP.Mur genotype.Due to ethnic integration,no significant difference was noticed in the frequency of Mur blood type distribution between Han,Zhuang and Yao population.Therefore,conducting extensive Mur blood group antigen and antibody testing in Hezhou is of great significance for ensuring clinical blood transfusion safety.

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