1.Effects of Modified Buyang Huanwu Tang on Mice with Cerebral Ischemia-reperfusion Injury by Regulating PINK1/Parkin Signaling Pathway-mediated Mitochondrial Autophagy
Li GUO ; Hengwen CHEN ; Cun ZHAN ; Zhenzhen YING ; Zuomin WU ; Shaoju JIN ; Shangmei CAO ; Shengming HUANG ; Jin WANG ; Xiaotao YU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(11):34-43
ObjectiveTo investigate the effects of modified Buyang Huanwu Tang on cerebral ischemia-reperfusion injury (CI/RI) in mice via the PTEN-induced putative kinase 1/E3 ubiquitin ligase (PINK1/Parkin) signaling pathway-mediated mitophagy, and to explore the underlying mechanism by which modified Buyang Huanwu Tang improves CI/RI. MethodsSeventy-two male C57BL/6J mice were randomly divided into six groups (n = 12 per group): Sham-operated group, middle cerebral artery occlusion/reperfusion (MCAO/R) model group, low-, medium-, and high-dose modified Buyang Huanwu Tang groups (8.84, 17.68, 35.36 g·kg-1·d-1), and an aspirin group (13.00 mg·kg-1·d-1). Neurological deficit scores were assessed using the Zea-Longa method. Cerebral infarct volume ratio was measured by 2,3,5-triphenyltetrazolium chloride (TTC) staining. Histopathological changes and neuronal injury in brain tissues were observed using hematoxylin-eosin (HE) staining and Nissl staining. Apoptosis was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) assay. Mitochondrial ultrastructure in brain tissue was observed by transmission electron microscopy (TEM). Serum levels of superoxide dismutase (SOD) and malondialdehyde (MDA) were determined by enzyme-linked immunosorbent assay (ELISA). The mRNA and protein expression levels of PINK1, Parkin, microtubule-associated protein 1 light chain 3B (LC3B, LC3Ⅱ/Ⅰ), and p62 in brain tissues were detected by real-time quantitative reverse transcription PCR (Real-time PCR) and Western blot, respectively. ResultsCompared with the sham-operated group, the MCAO/R model group showed significantly increased neurological deficit scores and cerebral infarct volume ratios (P<0.01). Severe cortical injury on the infarct side was observed, characterized by decreased neuronal density, cytoplasmic vacuolation, nuclear pyknosis, a marked reduction in Nissl bodies, dissolution of Nissl bodies in the cytoplasm of some pyramidal neurons, and blurred cellular boundaries. The number of TUNEL-positive cells increased significantly (P<0.01). Mitochondria exhibited cristae membrane rupture and matrix vacuolation, with rupture of the outer mitochondrial membrane and formation of autophagosomes, the number of which increased significantly. Serum SOD activity decreased significantly (P<0.01), while MDA content increased significantly (P<0.01). In infarcted brain tissues of model mice, the relative mRNA expression and protein levels of PINK1, Parkin and LC3B were significantly increased (P<0.05, P<0.01), whereas p62 mRNA and protein expression were significantly decreased (P<0.05, P<0.01), showing statistical significance. Compared with the model group, all treatment groups showed significantly decreased neurological deficit scores and cerebral infarct volume ratios (P<0.01). Neuronal density increased significantly, cytoplasmic vacuolation was alleviated, nuclear morphology tended to be more regular and clearer, Nissl body density increased significantly with reduced dissolution and improved contour clarity. The mitochondrial cristae structure was partially restored, with some mitochondria showing autophagosome encapsulation, and the degree of mitochondrial damage was alleviated. Serum SOD activity increased significantly (P<0.01), while MDA content decreased significantly. The mRNA and protein expression levels of PINK1, Parkin, and LC3Ⅱ/Ⅰ were significantly increased (P<0.05, P<0.01), while p62 mRNA and protein expression in the low- and medium-dose modified Buyang Huanwu Tang groups were significantly decreased (P<0.05, P<0.01), showing statistical significance. ConclusionModified Buyang Huanwu Tang can upregulate the protein expression levels of PINK1, Parkin, and LC3Ⅱ/Ⅰ and downregulate p62 protein expression, suggesting that it may improve CI/RI by regulating the expression of proteins related to the PINK1/Parkin signaling pathway. Regulation of the mitophagy pathway may be one of the mechanisms by which modified Buyang Huanwu Tang alleviates CI/RI in mice.
2.Variations of Chemical Components in Gardeniae Fructus Before and After Being Charred Analyzed by UPLC-Q-Orbitrap MS/MS
Lan LI ; Jie HONG ; Yanan SONG ; Yilan LI ; Yun WANG ; Cun ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(4):175-182
ObjectiveTo investigate the changes in chemical components of Gardeniae Fructus(GF) before and after being charred, providing data support for research on the material basis of GF Carbonisata(GFC). MethodsUltra-performance liquid chromatography-quadrupole-electrostatic field orbitrap high-resolution mass spectrometry(UPLC-Q-Orbitrap MS/MS) was used to conduct a comprehensive analysis of the chemical components in GF and GFC under positive and negative ion modes with Compound Discoverer 3.3 software and online database. Then, principal component analysis and partial least squares-discriminant analysis in SIMCA14.1 software were used to analyze the MS data of each sample. Based on the principle of variable importance in the projection(VIP) value>1, differential secondary and primary metabolites before and after carbonization were screened. In addition, MetaboAnalyst website was used for pathway enrichment of Kyoto Encyclopedia of Genes and Genomes(KEGG), so as to provide a reference for clarifying the processing mechanism. ResultsA total of 185 components were identified, including 96 secondary metabolites and 89 primary metabolites. These components were classified into nine categories, primarily including iridoid glycosides, flavonoids, and terpenoids, their fragmentation pathways were also analyzed. Simultaneously, multivariate statistical analysis was performed on the secondary and primary metabolites, identifying 70 and 59 differential metabolites, respectively. The secondary metabolites were enriched in two metabolic pathways, including C5-branched dibasic acid metabolism and flavonoid and flavonol biosynthesis, while the primary differential metabolites were enriched in seven pathways such as linoleic acid metabolism and tyrosine metabolism. ConclusionThe chemical components of GF change significantly after carbonization, with a significant decrease in the contents of iridoid glycosides and terpenoids such as hydroxyisogeniposide, crocin Ⅱ, crocetin, and jasminoside B. while the contents of 4-hydroxycoumarin, geniposidic acid, gentiopicroside, and gardenoside methyl ester increase significantly. This change is presumed to be associated with the enhanced cooling and hemostatic effects of the processed products. The identified key components provide a basis for elucidating the material basis underlying the efficacy changes before and after carbonization.
3.Analysis on Dynamic Change of Stir-fried Glycyrrhizae Radix et Rhizoma Quality Based on "Exterior-interior Correlation"
Yue XU ; Zhe JIA ; Yun WANG ; Bing LI ; Deling WU ; Cun ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(5):194-202
ObjectiveIn order to provide a reference for the optimization of preparation process of stir-fried Glycyrrhizae Radix et Rhizoma(sf-GRR), the quality changes during the processing was studied. MethodsGlycyrrhizae Radix et Rhizoma was processed by stir-frying for 17 min, and samples were collected every 1 min during the processing. The appearance color of the samples was determined by visual analysis technology, the moisture and extract of the process samples were detected by the drying method and the hot extraction method of alcohol-soluble extract in the general rules of the 2020 edition of Chinese Pharmacopoeia(part Ⅳ), and the contents of liquiritin apioside, liquiritin, isoliquiritin apioside, isoliquiritin, licoricesaponin G2 and glycyrrhizic acid in the process samples were determined by high performance liquid chromatography(HPLC). Then principal component analysis(PCA), partial least squares-discriminant analysis(PLS-DA) and Spearman correlation analysis were used for clustering, discrimination and correlation analysis of the appearance color, moisture, extract and the contents of six internal components. Based on artificial neural network and random forest algorithm, the prediction model of processing degree of sf-GRR was established. On this basis, based on the five principles of quality marker(Q-Maker), explore the monitoring Q-Maker of sf-GRR. ResultsThe color of Glycyrrhizae Radix et Rhizoma deepened after stir-frying, and the appearance color of the sample changed from light yellow to dark yellow during processing. During the stir-frying process, the moisture content showed a decreasing trend with the extension of processing time, while the extract content showed an increasing trend with the extension of processing time. After stir-frying, the contents of liquiritin apioside, liquiritin and licoricesaponin G2 showed an overall decreasing trend, while the contents of isoliquiritin apioside and isoliquiritin increased, and the content of glycyrrhizic acid increased slightly. The correlation analysis showed that moisture was positively correlated with brightness(L*) and red/green value(a*), and negatively correlated with yellow/blue value(b*) and total color difference(E*ab). Isoliquiritin apioside and isoliquiritin had negative correlation with L* and a*, and positive correlation with b* and E*ab. The processing process of sf-GRR could be divided into two stages of the early stage(0-14 min) and the late stage(15-17 min), and could be divided into three stages of the early stage(0-6 min), the middle stage(7-14 min) and the late stage(15-17 min) by combining the moisture, extract, the contents of 6 components and color values. Based on artificial neural network analysis and random forest algorithm, isoliquiritin apioside, isoliquiritin, liquiritin and glycyrrhizic acid were selected as monitoring markers for sf-GRR. ConclusionBased on the analysis of the exterior-interior indicators of process samples of sf-GRR, this paper ultimately identifies four processing monitoring markers, which can provide a basis for optimizing the processing technology of sf-GRR.
4.Processing History and Modern Research of Jianghuanglian: A Review
Ying LI ; Yun WANG ; Zhe JIA ; Lin YAN ; Min JIN ; Cun ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(6):275-282
Jianghuanglian is one of the representative processed products of Coptidis Rhizoma for treating cold syndrome with drugs of heat nature, and ginger is used to restrict the bitter cold of Coptidis Rhizoma, which can be traced back to Bojifang, and it is suitable for stagnation of damp-heat in middle-jiao, cold-heat mutual knots and other symptoms. Jianghuanglian retains the alkaloids, phenylpropanoids and flavonoids of Coptidis Rhizoma, and also introduces gingerol components such as 6-gingerol in ginger, which has pharmacological activities such as anti-inflammatory, antibacterial, anti-tumor, and improving gastrointestinal function. The 2020 edition of Chinese Pharmacopoeia and many local processing specifications have included the traditional processing process and quality standards of Jianghuanglian, but the specific process parameters and quality standards are incomplete, which limits the production and clinical application of this processed product. By summarizing the processing history, process research, quality evaluation, pharmacodynamic and medicinal property changes and application of Jianghuanglian in the past 20 years, there are differences in the processing methods and standards in various provinces and cities, which are mainly reflected in the preparation method, dosage, processing process and quantitative standards of ginger juice. In addition, there are also certain differences in the changes of the main components of Jianghuanglian prepared from ginger or dried ginger, as well as their efficacy and medicinal properties. The research on the processing process of Jianghuanglian plays an important role in improving its quality standards, and this review can provide a reference for improving the quality evaluation system of Jianghuanglian.
5.SITA:Predicting site-specific immunogenicity for therapeutic antibodies
Yewei CUN ; Hao DING ; Tiantian MAO ; Yuan WANG ; Caicui WANG ; Jiajun LI ; Zihao LI ; Mengdie HU ; Zhiwei CAO ; Tianyi QIU
Journal of Pharmaceutical Analysis 2025;15(6):1378-1389
Antibody humanization is critical to reduce immunogenicity and enhance efficacy in the preclinical phase of the development of therapeutic antibodies originated from animal models.Computational suggestions have long been desired,but available tools focused on immunogenicity calculation of whole antibody sequences and sequence segments,missing the individual residue sites.This study introduces Site-specific Immunogenicity for Therapeutic Antibody(SITA),a novel computational framework that predicts B-cell immunogenicity score for not only the overall antibody,but also individual residues,based on a comprehensive set of amino acid descriptors characterizing physicochemical and spatial features for antibody structures.A transfer-learning-inspired framework was purposely adopted to overcome the scarcity of Antibody-Antibody structural complexes.On an independent testing dataset derived from 13 Antibody-Antibody structural complexes,SITA successfully predicted the epitope sites for Antibody-Antibody structures with a receiver operating characteristic(ROC)-area unver the ROC curve(AUC)of 0.85 and a precision-recall(PR)-AUC of 0.305 at the residue level.Furthermore,the SITA score can significantly distinguish immunogenicity levels of whole human antibodies,therapeutic antibodies and non-human-derived antibodies.More importantly,analysis of an additional 25 thera-peutic antibodies revealed that over 70%of them were detected with decreased immunogenicity after modification compared to their parent variants.Among these,nearly 66%antibodies successfully iden-tified actual modification sites from the top five sites with the highest SITA scores,suggesting the ability of SITA scores for guide the humanization of antibody.Overall,these findings highlight the potential of SITA in optimizing immunogenicity assessments during the process of therapeutic antibody design.
6.Correlation Analysis of Northwest Dryness Syndrome with Pulmonary Tuberculosis in Moyu County,Hetian Prefecture,Xinjiang
Xiu-mei WANG ; Huan LIU ; Yu-cun ZHANG ; GULISHALA·TURSUN ; Hua-gui LI
Progress in Modern Biomedicine 2025;25(11):1788-1798
Objective:To explore the incidence characteristics and correlation analysis of Western dryness syndrome in pulmonary tuberculosis patients in Moyu County,Hetian Prefecture,Xinjiang,and to provide a basis for the prevention and treatment strategies and effectiveness evaluation of pulmonary tuberculosis in Xinjiang.Methods:89 confirmed cases of pulmonary tuberculosis in Moyu County,Hotian Prefecture,Xinjiang from October 2023 to October 2024 were selected as the case group,and 117 healthy individuals matched with the case group in terms of gender,age,and education level in Moyu County,Hotian Prefecture,Xinjiang during the same period were selected as the control group.The general information and incidence of Northwest dryness syndrome between two groups were compared.The incidence and severity of Northwest dryness syndrome in tuberculosis patients,as well as the main and secondary syndrome scores of Northwest dryness syndrome were analyzed.The influencing factors of pulmonary tuberculosis were analyzed.The scores of the main syndrome of Northwest dryness syndrome(lung wei kong pi dryness syndrome)and the concurrent syndrome of Northwest dryness syndrome(heart kidney yin deficiency syndrome,lung heart spleen wind fire dryness syndrome,liver kidney essence blood deficiency syndrome,spleen stomach dampness syndrome,spleen stomach yin deficiency syndrome)between the control group and the case group were compared.The correlation between pulmonary tuberculosis and the on set of Northwest dryness syndrome were analyzed by Spearman rank sum test.Results:There was no significant difference between the control group and the case group in terms of body mass index(BMI),age,education level,gender,etc.(P>0.05).The incidence of Northwest dryness syndrome in the case group was significantly higher than that in the control group(P<0.05).The Northwest Dry Syndrome Scale score in the severe group was higher than that in the mild and moderate groups,and the Northwest Dry Syndrome Scale score in the moderate group was higher than that in the mild group(P<0.05).Among 89 patients with pulmonary tuberculosis,the main syndrome of lung wei kong pi dryness syndrome had the highest number of cases,totaling 42 cases(47.19%).The highest number of concurrent syndromes were heart kidney yin deficiency syndrome,totaling 36 cases(40.45%),followed by spleen stomach yin deficiency syndrome,totaling 35 cases(39.33%).The results showed that suffering from Northwest dryness syndrome was a risk factor for pulmonary tuberculosis(P<0.05).There were significant differences in the scores of the main syndrome of Northwest dryness(Lung Wei Kong Pi dryness syndrome),the concurrent syndrome of Northwest dryness(Lung Heart Spleen Wind Fire dryness syndrome),and the concurrent syndrome of Northwest dryness(Spleen Stomach Yin Deficiency syndrome)between the control group and the case group(P<0.05).There was no significant difference in the scores of Northwest dryness syndrome(liver and kidney essence and blood deficiency syndrome),Northwest dryness syndrome(heart and kidney yin deficiency syndrome),and Northwest dryness syndrome(spleen and stomach dampness syndrome)between the control group and the case group(P>0.05).The score of pulmonary tuberculosis syndrome was positively correlated with various types of Northwest dryness syndrome(P<0.05),and there was a significant correlation at the 0.001 level between pulmonary Wei Kong Pi dryness syndrome,pulmonary heart spleen wind fire dryness syndrome,and spleen stomach dampness syndrome(P<0.05).The correlation between liver and kidney essence and blood deficiency syndrome and spleen and stomach yin deficiency syndrome is significant at the 0.01 level(P<0.05).The correlation between heart and kidney yin deficiency syndrome is significant at the 0.05 level(P<0.05).Conclusion:The northwest dryness syndrome is related to the occurrence of pulmonary tuberculosis in Moyu County,Hotan Prefecture,Xinjiang,and is a risk factor for the occurrence of pulmonary tuberculosis.There is a significant positive correlation between the pulmonary tuberculosis syndrome score and various types of northwest dryness syndrome,among which the lung wei kong pi dryness syndrome,lung heart spleen wind fire dryness syndrome,and spleen stomach dampness syndrome have the most significant impact on the pulmonary nucleus syndrome score.
7.Mechanism of Ganoderma lucidum polysaccharides promoting myelin regeneration in demyelinated mice induced by cuprizone
Yan-qing LI ; Xiao-hui LI ; Qing WANG ; Li-juan SONG ; Li-zhi YANG ; Han-bin WANG ; Bao-guo XIAO ; Cun-gen MA
Chinese Pharmacological Bulletin 2025;41(7):1265-1273
Aim To explore the mechanism of Gano-derma lucidum polysaccharides(GLPS)promoting my-elin repair and regeneration in mice with chronic demy-elination induced by cuprizone(CPZ).Methods A total of 40 C57BL/6 mice were randomly divided into four groups:Normal+NS,Normal+GLPS,CPZ+NS and CPZ+GLPS.A chronic demyelination model was established using 0.2%CPZ.Open field and elevated plus maze tests were performed to observe the behavior-al changes in the mice.Immunofluorescence staining and Western blot were used to detect changes in myelin basic protein expression in the corpus callosum.ELISA was performed to measure the levels of TNF-α,IL-6,IL-1β and IL-10 in brain homogenates.Immunofluo-rescence staining was also used to observe the expres-sion of ionized calcium binding adapter molecule 1(Iba1)and neural-glial antigen 2(NG2).RT-qPCR and Western blot were conducted to assess the mRNA and protein expression levels of MBP,iNOS,COX-2,JAK2 and STAT3.Results Mice in the CPZ+NS group showed a significant decrease in body weight,cognitive behavior abnormalities,and impaired myelin regeneration.The expression of pro-inflammatory fac-tors increased,while anti-inflammatory factors de-creased.Additionally,Iba1 and NG2 expression in-creased,and the JAK2/STAT3 signaling pathway was activated.After GLPS intervention,the mouse body weight increased,myelin regeneration occurred,cogni-tive behavior was improved,the expression of inflamma-tory factors decreased,anti-inflammatory factors in-creased,NG2 expression was further elevated,and the proliferation of microglia as well as the activation of the JAK2/STAT3 signaling pathway was inhibited.Conclu-sions GLPS can improve cognitive behavior abnormali-ties and inflammatory responses in chronic demyelinated mice by inhibiting the JAK2/STAT3 signaling pathway,thereby promoting myelin repair and regeneration.
8.Establishment of near-infrared spectroscopy quantitative models for moisture and index components in Alismatis Rhizoma decoction pieces
Xun LU ; Zhe ZHANG ; Geng-zhi ZHAN ; Lu-yao CAI ; Cun-yu LI ; Yun-feng ZHENG ; Tuan-jie WANG ; Yu JIN ; Guo-ping PENG
Chinese Traditional Patent Medicine 2025;47(10):3184-3190
AIM To establish the near-infrared spectroscopy quantitative models for moisture,23-acetylalismol B and 23-acetylalismol C in Alismatis Rhizoma decoction pieces.METHODS The near-infrared spectroscopy(NIRS)data were collected in 95 batches of decoction pieces,after which drying method was adopted in the content determination of moisture,HPLC was applied to determining the contents of 23-acetylalismol B and 23-acetylalismol C,the quantitative models were established by partial least squares method combined with feature extraction algorithms.RESULTS The model training determination coefficients were 0.952 6,0.958 1 and 0.920 8,along with the prediction determination coefficients of 0.930 0,0.905 2 and 0.906 4,the residual prediction deviations(PRD)of 4.00,3.58 and 3.46,and the root mean square error ratios of prediction values to calibration values(RMSEP/RMSEC)of 1.15,1.11 and 1.06,respectively.CONCLUSION The quantitative models based on NIRS exhibit good prediction effects,which can be used for the rapid quality detection of Alismatis Rhizoma decoction pieces.
9.Application of biomechanical simulation based on three-dimensional human body model in preventing pressure ulcers:a scoping review
Lingxiao RAN ; Dongmin WANG ; Ke XU ; Cong WANG ; Hua CAO ; Wei CUN ; Yan JIANG
Chinese Journal of Nursing 2025;60(8):1012-1018
Objective A scoping review of domestically and internationally published studies on finite element analysis(FEA)based on three-dimensional(3D)human body model in preventing pressure ulcer(PU)was conducted,aiming to provide new directions for improving the prevention strategies of pressure ulcer.Methods We conducted a systematic search in both Chinese and English medical databases,including PubMed,Embase,CINAHL,Web of Science,Cochrane Library,Sinomed,CNKI,Wanfang and VIP,and engineering database(Engineering Village Compen-dex).The search period was from the inception of each database to July 29,2024.The information was extracted,and the results were analyzed and standardized for reporting.Results A total of 30 studies were included.The general methods of FEA based on 3D human body model include establishing geometric model,meshing,defining material properties,loading and setting boundary conditions,and solving equations.According to the contents of studies,they could be categorized into 4 distinct application domains,including identification of risk groups(n=9),position management(n=9),preventive dressings(n=10)and supportive surfaces(n=7).Conclusion FEA based on 3D human body model provides a foundation to PU biomechanical mechanism research and a scientific basis to the supplement and optimization of clinical prevention.Future studies should integrate clinical problems with simulations and further optimize simulations techniques and protocols.
10.Comparative study of combined external fixator and plate fixation in the treatment of humerus throwing fracture
Pei-gang GAO ; Xing-cun WANG ; Li-qiang ZHU
China Journal of Orthopaedics and Traumatology 2025;38(6):613-618
Objective To compare clinical efficacy of combined external fixator and plate fixation in treating throwing fracture of humerus.Methods Sixty-seven male patients with throwing humerus fracture admitted from February 2018 to February 2021 were retrospectively analyzed and divided into external fixator group and plate group according to different treatment methods.There were 32 patients in external fixator group,aged from 18 to 27 years old with an average of(23.6±2.6)years old;19 patients with type A,13 patients with type B according to AO classification;fixed by combined external fixer.There were 35 patients in plate group,aged from 17 to 28 years old with an average of(23.2±2.9)years old;21 patients with type A,14 patients with type B according to AO classification;fixed with steel plate screws.Operation time,intraoperative blood loss,total incision length,fracture healing time and complications were compared between two groups.Clinical effect was evaluated by Mayo elbow performance score(MEPS)and shoulder Neer score at 12 months after operation.Results Patients in both groups were successfully completed operation and followed up for 15 to 21 months with an average of(18.3±3.4)months.In external fixator group,intraoperative blood loss,incision length,fracture healing time and postoperative MEPS of el-bow joint function at 12 months were(68.5±15.7)ml,(4.9±1.2)cm,(10.5±2.9)weeks,(93.6±5.1)min,respectively;which were superior to ml(149.4±38.9)ml,(12.5±2.4)cm,(12.8±2.7)weeks,(85.6±6.8)in plate group(P<0.05),and had signif-icant differences;there were no significant difference in operative time and postoperative Neer score of shoulder joint function at 12 months between two groups(P>0.05).There were 6 patients occurred radial nerve injury in plate group and no patient occurred radial nerve injury in external fixator group,and the difference between two groups was statistically significant(x2=6.025,P<0.05).There were no significant difference in postoperative MEPS grading and Neer shoulder joint grading at 12 months(P>0.05).Bone healing was achieved in both groups without failure of internal fixation.Conclusion The combined ex-ternal fixator for the treatment of humerus throwing fracture has characteristics of low incidence of radial nerve injury,less bleeding,less complications,fast fracture healing,which is convenient to remove after fracture healing.

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