1.Wen-Shen-Tong-Du Decoction promoting spinal cord injury repair in mice
Ruihua ZHAO ; Sixian CHEN ; Yang GUO ; Lei SHI ; Chengjie WU ; Mao WU ; Guanglu YANG ; Haoheng ZHANG ; Yong MA
Chinese Journal of Tissue Engineering Research 2025;29(6):1118-1126
BACKGROUND:Previous studies have confirmed that Wen-Shen-Tong-Du Decoction can promote the recovery of spinal cord injury by inhibiting pyroptosis of splenic B cells,promoting the phagocytosis of myelin debris by microvascular endothelial cells,affecting the migration and infiltration of microglia,promoting the recovery of damaged neurons,and decreasing neuronal apoptosis after spinal cord injury,but the mechanism of this is still not clear. OBJECTIVE:To investigate the effect of Wen-Shen-Tong-Du Decoction on the triggering receptor expressed on myeloid cells 2(TREM2)and PI3K/Akt signaling pathways in mice following spinal cord injury. METHODS:Thirty-six C57BL/6 mice were selected and randomly divided into a sham-operation group,a model group and a Wen-Shen-Tong-Du Decoction group,with 12 mice in each group.In the model and Wen-Shen-Tong-Du Decoction groups,mouse models of T10 spinal cord injury were prepared by the modified Allen's method.On the 1st day after modeling,the Wen-Shen-Tong-Du Decoction group was given Wen-Shen-Tong-Du Decoction by gavage,and the sham-operation group and the model group were given saline by gavage once a day for 28 days.During the drug administration period,mouse motor function was evaluated by Basso Mouse Scale score and inclined plane test.On the 7th and 28th days after modeling,hematoxylin-eosin staining was used to observe the histopathological changes in the spinal cord tissue of the mice;immunofluorescence double staining was used to detect the protein expression of ionized calcium binding adaptor molecule 1(IBA1)and TREM2;and western blot assay was used to detect the expression of TREM2,PI3K,p-PI3K,Akt,p-Akt,Bcl2,Bax and Caspase3 in spinal cord tissue. RESULTS AND CONCLUSION:Basso Mouse Scale scores and inclined plane test results indicated that the motor function of the mouse hindlimbs was declined after spinal cord injury,and Wen-Shen-Tong-Du Decoction significantly improved motor function in mice with spinal cord injury.Hematoxylin-eosin staining results revealed that Wen-Shen-Tong-Du Decoction significantly ameliorated the pathological structure of spinal cord tissue compared with the model group,manifesting as reduced degrees of dorsal white matter and neuronal atrophy,decreased cytoplasmic vacuolization,and reduced inflammatory cell infiltration.Immunofluorescence double staining results showed that on the 7th day after modeling,the protein expression of IBA1 and TREM2 in the model group was lower than that in the sham-operation group(P<0.05),and the protein expression of IBA1 and TREM2 in the Wen-Shen-Tong-Du Decoction group was higher than that in the model group(P<0.05);on the 28th day after modeling,the protein expression of TREM2 in the model group was lower than that in the sham-operation group(P<0.05),and the protein expression of TREM2 in the spinal cord tissue of the mice in the Wen-Shen-Tong-Du Decoction group was higher than that in the model group(P<0.05).Western blot results analysis demonstrated that on the 7th day after modeling,compared with the sham-operation group,the model group exhibited a significant reduction in TREM2,PI3K,and Bcl2/Bax(P<0.05),as well as a significant increase in p-Akt,Bax and p-Akt/Aktp-PI3K(P<0.05);compared with the model group,the Wen-Shen-Tong-Du Decoction group showed a significant increase in TREM2,PI3K,p-PI3K,Akt,p-Akt,Bcl2,p-PI3K/PI3K,p-Akt/Ak,and Bcl2/Bax(P<0.05),as well as a significant decrease in Bax and Caspase3 protein expression(P<0.05).On the 28th day after modeling,compared with the sham-operation group,the model group exhibited a significant reduction in TREM2,PI3K,p-PI3K,Akt,p-Akt,Bcl2 and Bcl2/Bax(P<0.05),as well as a significant increase in Bax protein expression(P<0.05);compared with the model group,the Wen-Shen-Tong-Du Decoction group showed a significant increase in TREM2,PI3K,Akt,p-Akt,Bcl2,and Bcl2/Bax(P<0.05),as well as a significant decrease in Bax protein expression(P<0.05).To conclude,Wen-Shen-Tong-Du Decoction may activate the PI3K/Akt signaling pathway by up-regulating the expression of TREM2 protein in microglia,and then inhibit neuronal apoptosis,thus exerting neuroprotective effects and promoting the repair of spinal cord injury.
2.Analysis of the clinical effect of tirofiban in the treatment of early neurological deterioration in patients with acute ischemic stroke
Xiaohui LI ; Xiaomin LI ; Mingyang WEI ; Huimin GUO ; Chen WANG ; Jianbin ZHANG ; Zhiqiang ZHAO
China Pharmacy 2025;36(10):1221-1225
OBJECTIVE To investigate the efficacy and safety of tirofiban for early neurological deterioration in patients with acute ischemic stroke. METHODS A total of 126 patients with early neurological deterioration of acute ischemic stroke who were admitted to the Department of Neurology, Heji Hospital Affiliated to Changzhi Medical College from January 2022 to December 2023 were selected and divided into observation group and control group according to random number table method, with 63 cases in each group. All patients received standardized treatment such as lipid-lowering and blood pressure-lowering therapy. Based on the standard treatment, patients in the control group additionally took Aspirin enteric-coated tablets 100 mg+Clopidogrel bisulfate tablets 75 mg orally (once a day, for 14 consecutive days). The patients in the observation group received Tirofiban hydrochloride and sodium chloride injection based on the standardized treatment [first intravenous infusion of 0.40 μg/(kg·min) for 30 min, and then continuous intravenous infusion of 0.10 μg/(kg·min) for 47.5 h]; subsequently, patients were given Aspirin enteric-coated tablets (100 mg) and Clopidogrel bisulfate tablets (75 mg) once a day for 14 consecutive days. The clinical efficacy, the National Institutes of Health Stroke Scale (NIHSS) score, modified Rankin Scale (mRS) score, and hemorheological indexes before and after treatment were compared between the two groups, and the adverse reactions were recorded. RESULTS The total effective rate (87.30%) of the observation group was significantly higher than that of the control group (71.43%) (P<0.05). NIHSS scores of the two groups at 1st, 7th and 14th day after treatment, the mRS score at 90th day after treatment, and the platelet aggregation rate, whole blood viscosity, plasma viscosity and fibrinogen at 14th day after treatment were significantly lower than those before treatment in the same group, and the observation group was significantly lower than the control group at the same period (P<0.05). The total incidences of adverse reactions such as nausea, headache, fever, gastrointestinal bleeding, oral and nasal mucosal bleeding and thrombocytopenia in both groups of patients were 28.57% respectively, with no statistically significant difference (P>0.05). CONCLUSIONS For patients with early neurological deterioration in acute ischemic stroke, the addition of tirofiban can accelerate the recovery of neurological function, improve blood hyperviscosity and platelet aggregation, and improve the prognosis of patients with good safety.
3.Study on the mechanism of rutin in ameliorating depressive symptoms associated with premenstrual dysphoric disorder characterized by liver qi stagnation syndrome
Yiwei ZHANG ; Xianliang SONG ; Yashuang REN ; Dedi GUO ; Runwei SONG ; Xitai CHEN ; Huaiwei ZHAO ; Chunhong SONG
China Pharmacy 2025;36(12):1449-1456
OBJECTIVE To investigate the mechanisms of rutin in alleviating depressive symptoms associated with premenstrual dysphoric disorder (PMDD) characterized by liver qi stagnation syndrome. METHODS Network pharmacology was employed to identify the intersecting targets of action between PMDD and rutin. A protein-protein interaction network was constructed to screen core targets, followed by gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis. Molecular docking simulations validated rutin’s binding affinity to core targets. The bilateral ovaries of female Wistar rats were removed, followed by artificial hormone induction. The rats were then randomly divided into normal group (10 rats) and modeling group (50 rats). PMDD rat model with liver qi stagnation syndrome was established via restraint stress. The successfully modeled rats were further divided into model group, fluoxetine group (positive control) and rutin group, with 12 rats in each group. The corresponding drug solutions or water were administered by gavage at 9:00 a.m. every day, continuing for two estrous cycles. The open-field test, forced swimming test and Y-maze test were utilized to evaluate the effects of rutin on the behavioral indexes of model rats. Additionally, the density of neuronal dendritic spines in the hippocampal tissues of the rats was observed. Serum brain-derived neurotrophic factor (BDNF) levels and the expressions of BDNF, tyrosine kinase receptor type B (TrkB), synuclein (Syn), and postsynaptic density protein 95 (PSD95) in hippocampal tissues were quantified, respectively. RESULTS Network pharmacology and molecular docking revealed the core targets through which rutin ameliorated PMDD characterized by liver qi stagnation syndrome included BDNF, TrkB, PSD65, Syn, etc. The results of experimental validation demonstrated that rutin significantly increased the spontaneous alternation behavior scores of PMDD model rats with liver qi stagnation syndrome during the non-receptive phase, shortened their immobility time during the forced swimming test, and enhanced the density of neuronal dendritic spines in the hippocampal tissues. Additionally, rutin upregulated the levels of serum BDNF and the protein expressions of BDNF, TrkB and Syn in the hippocampal tissues (P<0.05). However, it had no significant effect on the above indexes in model rats during the receptive phase (P>0.05). CONCLUSIONS Rutin ameliorates depressive symptoms, enhances spatial memory capabilities, and reduces neuronal damage in PMDD model rats with liver qi stagnation syndrome. These effects may be associated with the activation of BDNF/TrkB signaling pathway and upregulation of Syn protein expression.
4.Production of GTKO pigs and kidney xenotransplantation from pigs to rhesus macaques
Yan WANG ; Yue CHANG ; Chang YANG ; Taiyun WEI ; Xiaoying HUO ; Bowei CHEN ; Jiaoxiang WANG ; Heng ZHAO ; Jianxiong GUO ; Hongfang ZHAO ; Xiong ZHANG ; Feiyan ZHU ; Wenmin CHENG ; Hongye ZHAO ; Kaixiang XU ; Ameen Jamal MUHAMMAD ; Zhendi WANG ; Hongjiang WEI
Organ Transplantation 2025;16(4):526-537
Objective To explore the construction of α-1,3-galactosyltransferase (GGTA1) gene-knockout (GTKO) Diannan miniature pigs and the kidney xenotransplantation from pigs to rhesus macaques, and to assess the effectiveness of GTKO pigs. Methods The GTKO Diannan miniature pigs were constructed using the CRISPR/Cas9 gene-editing system and somatic cell cloning technology. The phenotype of GTKO pigs was verified through polymerase chain reaction, Sanger sequencing and immunofluorescence staining. Flow cytometry was used to detect antigen-antibody (IgM) binding and complement-dependent cytotoxicity. Kidney xenotransplantation was performed from GTKO pigs to rhesus macaques. The humoral immunity, cellular immunity, coagulation and physiological indicators of the recipient monkeys were monitored. The function and pathological changes of the transplanted kidneys were analyzed using ultrasonography, hematoxylin-eosin staining, immunohistochemical staining and immunofluorescence staining. Results Single-guide RNA (sgRNA) targeting exon 4 of the GGTA1 gene in Diannan miniature pigs was designed. The pGL3-GGTA1-sgRNA1-GFP vector was transfected into fetal fibroblasts of Diannan miniature pigs. After puromycin selection, two cell clones, C59# and C89#, were identified as GGTA1 gene-knockout clones. These clones were expanded to form cell lines, which were used as donor cells for somatic cell nuclear transfer. The reconstructed embryos were transferred into the oviducts of trihybrid surrogate sows, resulting in 13 fetal pigs. Among them, fetuses F04 and F11 exhibited biallelic mutations in the GGTA1 gene, and F04 had a normal karyotype. Using this GTKO fetal pig for recloning and transferring the reconstructed embryos into the oviducts of trihybrid surrogate sows, seven surviving piglets were obtained, all of which did not express α-Gal epitope. The binding of IgM from the serum of rhesus monkey 20# to GTKO pig PBMC was reduced, and the survival rate of GTKO pig PBMC in the complement-dependent cytotoxicity assay was higher than that of wild-type pig. GTKO pig kidneys were harvested and perfused until completely white. After the left kidney of the recipient monkey was removed, the pig kidney was heterotopically transplanted. Following vascular anastomosis and blood flow restoration, the pig kidney rapidly turned pink without hyperacute rejection (HAR). Urine appeared in the ureter 6 minutes later, indicating successful kidney transplantation. The right kidney of the recipient was then removed. Seven days after transplantation, the transplanted kidney had good blood flow, the recipient monkey's serum creatinine level was stable, and serum potassium and cystatin C levels were effectively controlled, although they increased 10 days after transplantation. Seven days after transplantation, the levels of white blood cells, lymphocytes, monocytes and eosinophils in the recipient monkey increased, while platelet count and fibrinogen levels decreased. The activated partial thromboplastin time, thrombin time and prothrombin time remained relatively stable but later showed an upward trend. The recipient monkey survived for 10 days. At autopsy, the transplanted kidney was found to be congested, swollen and necrotic, with a small amount of IgG deposition in the renal tissue, and a large amount of IgM, complement C3c and C4d deposition, as well as CD68+ macrophage infiltration. Conclusions The kidneys of GTKO Diannan miniature pigs may maintain normal renal function for a certain period in rhesus macaques and effectively overcome HAR, confirming the effectiveness of GTKO pigs for xenotransplantation.
5.Association between screen time and anxiety-depression symptoms and their comorbidity among middle school students in Taiyuan City
Chinese Journal of School Health 2025;46(7):980-984
Objective:
To investigate the association between screen time (ST) during leisure time and anxiety-depression symptoms among middle school students, so as to provide a basis for formulating relevant intervention measures.
Methods:
From November to December 2023, a stratified cluster random sampling method was used to select 2 542 students from junior and senior high school in Taiyuan City. A self designed questionnaire, the Generalized Anxiety Disorder Scale (GAD-7), and the Patient Health Questionnaire (PHQ-9) were used to investigate ST and anxiety/depression symptoms among middle school students. The Logistic regression model was used to explore the association of ST with symptoms of anxiety and depression, as well as with anxiety and depression comorbiditles (CAD).
Results:
The detection rates of anxiety symptoms, depression symptoms, and CAD were 13.69%, 15.77%, and 10.11%, respectively. The median ST was 2.00 h/d [interquartile range ( IQR =2.38) for weekly averages], with 0.33 h/d ( IQR =1.67) on work days and 5.00 h/d ( IQR=5.50) on rest days. Logistic regression analysis indicated that the total ST of mobile phones during rest days ( OR =1.07, 1.10, 1.11) and the averages ST of mobile phones over a week ( OR = 1.20 , 1.22, 1.29), as well as the total ST of all screen types during rest days ( OR =1.04, 1.04, 1.05) and the averages ST of all screen types over a week ( OR =1.08, 1.09, 1.21) were positively correlated with anxiety symptoms, depression symptoms, and CAD (all P <0.01).
Conclusions
Among middle school students in Taiyuan City, screen time is positively correlated with symptoms of anxiety or depression and the comorbidity of anxiety and depression, especially smartphone screen time and weekend screen use. Therefore, measures should be implemented to reduce unnecessary screen time among middle school students, especially the use of mobile phones, in order to mitigate the occurrence of anxiety and depression.
6.Construction and Functional Validation of GTKO/hCD55 Gene-Edited Xenotransplant Donor Pigs
Jiaoxiang WANG ; Lu ZHANG ; Shuhan CHEN ; Deling JIAO ; Heng ZHAO ; Taiyun WEI ; Jianxiong GUO ; Kaixiang XU ; Hongjiang WEI
Laboratory Animal and Comparative Medicine 2025;45(4):379-392
Objective To develop GTKO (α-1,3-galactosyltransferase gene-knockout, GTKO)/hCD55 (human CD55) gene-edited xenotransplant donor pigs and verify their function. Methods In this study, CRISPR (clustered regularly interspaced short palindromic repeats)/Cas9 (CRISPR-associated nuclease 9), PiggyBac transposon technology and somatic cell nuclear transfer technology were used to construct GTKO/hCD55 gene-edited Diannan miniature pigs. The phenotype and function of GTKO/hCD55 pigs were analyzed by Sanger sequencing, real-time fluorescence quantitative PCR, flow cytometry, immunofluorescence, bisulfite sequencing, antigen-antibody binding assays, and complement-dependent cytotoxicity assays. Results After transfection of PX458 and PiggyBac gene editing vectors into wild-type fetal pig fibroblasts, 48 single-cell colonies were obtained through puromycin drug screening. Two single-cell colonies were selected for somatic cell nuclear transfer, resulting in two fetal pigs at 33 days of gestation. The GGTA1(α-1,3-galactosyltransferase) genotypes of fetal pig F01 were -17 bp and wild type (WT), while the GGTA1 genotypes of fetal pig F02 were -26 bp/+2 bp and -3 bp. The hCD55 mRNA expression levels of both fetal pigs were significantly higher than those of WT pigs (P<0.01). The fetal pig F02 was selected as the donor cell source for recloning, 11 surviving piglets were obtained, all identified as GTKO/hCD55 gene-edited pigs. These pigs showed absence of α-Gal antigen expression, but weak or no expression of hCD55 was observed. Methylation analysis of the hCD55 gene's CpG island showed hypermethylation in kidney tissue lacking hCD55 expression, whereas it was not methylated or partially methylated in kidney tissue expressing hCD55. Moreover, codon optimization of the CpG island of the hCD55 gene to reduce CG content could achieve stable expression of the hCD55 gene. In addition, antigen-antibody binding experiment showed that the amount of human IgM binding to GTKO/hCD55 gene-edited pig fibroblasts was significantly lower than that of WT pigs (P<0.01). Complement-dependent cytotoxicity experiment showed that the survival rate of fibroblasts in GTKO/hCD55 pigs was significantly higher than that in WT pigs (P<0.01). Conclusion This study demonstrates the successful generation of GTKO/hCD55 gene-edited xenotransplant donor pigs. Methylation-induced gene silencing of the hCD55 gene can be effectively avoided by reducing the CG content of the CpG island through codon optimization. This study provides a reference for the development of xenotransplant donor pigs and guides subsequent research on xenotransplantation.
7.Artemisia argyi extract subfraction exerts an antifungal effect against dermatophytes by disrupting mitochondrial morphology and function.
Le CHEN ; Yunyun ZHU ; Chaowei GUO ; Yujie GUO ; Lu ZHAO ; Yuhuan MIAO ; Hongzhi DU ; Dahui LIU
Chinese Journal of Natural Medicines (English Ed.) 2024;22(1):47-61
Artemisia argyi (A. argyi), a plant with a longstanding history as a raw material for traditional medicine and functional diets in Asia, has been used traditionally to bathe and soak feet for its disinfectant and itch-relieving properties. Despite its widespread use, scientific evidence validating the antifungal efficacy of A. argyi water extract (AAWE) against dermatophytes, particularly Trichophyton rubrum, Trichophyton mentagrophytes, and Microsporum gypseum, remains limited. This study aimed to substantiate the scientific basis of the folkloric use of A. argyi by evaluating the antifungal effects and the underlying molecular mechanisms of its active subfraction against dermatophytes. The results indicated that AAWE exhibited excellent antifungal effects against the three aforementioned dermatophyte species. The subfraction AAWE6, isolated using D101 macroporous resin, emerged as the most potent subfraction. The minimum inhibitory concentrations (MICs) of AAWE6 against T. rubrum, M. gypseum, and T. mentagrophytes were 312.5, 312.5, and 625 μg·mL-1, respectively. Transmission electron microscopy (TEM) results and assays of enzymes linked to cell wall integrity and cell membrane function indicated that AAWE6 could penetrate the external protective barrier of T. rubrum, creating breaches ("small holes"), and disrupt the internal mitochondrial structure ("granary"). Furthermore, transcriptome data, quantitative real-time PCR (RT-qPCR), and biochemical assays corroborated the severe disruption of mitochondrial function, evidenced by inhibited tricarboxylic acid (TCA) cycle and energy metabolism. Additionally, chemical characterization and molecular docking analyses identified flavonoids, primarily eupatilin (131.16 ± 4.52 mg·g-1) and jaceosidin (4.17 ± 0.18 mg·g-1), as the active components of AAWE6. In conclusion, the subfraction AAWE6 from A. argyi exerts antifungal effects against dermatophytes by disrupting mitochondrial morphology and function. This research validates the traditional use of A. argyi and provides scientific support for its anti-dermatophytic applications, as recognized in the Chinese patent (No. ZL202111161301.9).
Antifungal Agents/chemistry*
;
Arthrodermataceae
;
Artemisia/chemistry*
;
Molecular Docking Simulation
;
Mitochondria
;
Microbial Sensitivity Tests
8.Mechanism of Medicated Serum of Buyang Huanwutang in Inhibiting Endothelial-to-mesenchymal Transition of Human Pulmonary Artery Endothelial Cells Based on Wnt1/β-catenin Signaling Pathway
Qifen YANG ; Jinglian QU ; Huiliang ZHAO ; Yongsheng GUO ; Long CHEN
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(7):49-58
ObjectiveTo investigate the role of the Wnt1/β-catenin signaling pathway in the intervention of medicated serum of Buyang Huanwutang (BYHWT) in endothelial-to-mesenchymal transition (EndMT) of human pulmonary artery endothelial cells (HPAECs) as well as its related mechanisms. MethodMedicated serum of BYHWT was prepared by gavage to New Zealand rabbits with a dosage of 53.36 g·kg-1·d-1 after decocting the medicine as usual. In addition, the same volume of normal saline was used to prepare blank serum. The HPAECs were cultured in vitro, and then induced by the transforming growth factor-β1 (TGF-β1) to establish the EndMT model. Five groups were established: blank group (10% blank serum), model group (TGF-β1+10% blank serum), low-dose BYHWT group (TGF-β1+2.5% medicated serum+7.5% blank serum), medium-dose BYHWT group (TGF-β1+5% medicated serum+5% blank serum) and high-dose BYHWT group (TGF-β1+10% medicated serum). Through Western blot, the expressions of Wnt1, β-catenin, and glycogen synthase kinase-3β (GSK-3β) were detected. In order to further clarify the mechanism of the Wnt1/β-catenin signaling pathway in the intervention of the medicated serum of BYHWT in inhibiting EndMT, the overexpression of β-catenin was confirmed by polymerase chain reaction after plasmid of overexpression β-catenin was constructed and transfected into the HPAECs. The HPAECs were intervened by 10% medicated serum with the optimal effect in previous studies. Then, they were divided into another five groups: the blank group (10% blank serum), the model group (TGF-β1+10% blank serum), the BYHWT group (TGF-β1+10% medicated serum), the BYHWT+overexpression plasmid control group (TGF-β1+10% medicated serum+blank plasmid) and the BYHWT+β-catenin overexpression plasmid group (TGF-β1+10% medicated serum+β-catenin). Apart from that, cell proliferation ability was detected by the methyl thiazolyl tetrazolium (MTT) method and cell migration ability by scratch assay and Transwell assay together. Immunofluorescence was adopted to detect the expressions of platelet endothelial cell adhesion molecule (PECAM-1/CD31), vascular endothelial cadherin (VE-cadherin), fibroblast-specific protein 1 (FSP1), and α-smooth muscle actin (α-SMA). ResultIn comparison to the blank group, the expressions of Wnt1 and β-catenin were significantly increased (P<0.01) while the expression of GSK-3β significantly decreased (P<0.01) in the model group. In comparison to the model group, the expressions of Wnt1 and β-catenin were significantly decreased (P<0.01) while the expression of GSK-3β was significantly increased (P<0.01) in the high-dose BYHWT group. The expression of β-catenin was significantly decreased (P<0.01) while the expression of GSK-3β was significantly increased (P<0.01) in the medium-dose BYHWT group. There was no significant difference in these indexes of the low-dose BYHWT group. In comparison to the blank group, proliferation and migration abilities were remarkably increased (P<0.01) and the immunofluorescence intensities of CD31 and VE-cadherin were decreased, while those of FSP1 and α-SMA were increased in the model group. In comparison to the model group, proliferation and migration abilities were significantly decreased (P<0.01) and the immunofluorescence intensities of CD31 and VE-cadherin were increased, while those of FSP1 and α-SMA diminished in the BYHWT group. Beyond that, the change trend of those indexes in the BYHWT+β-catenin overexpression plasmid group was consistent with that in the model group. In comparison to the BYHWT+overexpression plasmid control group, proliferation and migration abilities were significantly increased (P<0.01) and the immunofluorescence intensities of CD31 and VE-cadherin were decreased, while those of FSP1 and α-SMA were increased in the BYHWT+β-catenin overexpression plasmid group. ConclusionMedicated serum of BYHWT can inhibit EndMT of HPAECs by the Wnt1/β-catenin signaling pathway.
9.Historical Evolution and Key Information Research on Pediatric Famous Classical Formula Yigongsan
Jiangmin SU ; Jun ZHANG ; Cong GUO ; Anyi ZHAO ; Liang JIANG ; Heng ZHANG ; Jipeng DI ; Sha CHEN ; Li LIU ; Yan LIU ; An LIU
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(7):205-214
Yigongsan is derived from Xiaoer Yaozheng Zhijue written by QIAN Yi in the Northern Song dynasty, which is the No. 3 formula in the Catalogue of Ancient Famous Classical Formulas(The Second Batch of Pediatrics) released by the National Administration of Traditional Chinese Medicine(TCM) in September 2022, and it can be developed as a class 3.1 new TCM drug. By referring to ancient medical books and modern literature, this study conducted herbal textual research on Yigongsan from five aspects, including historical evolution, origin and processing, dosage conversion, usage and preparation methods, and functional application, then formed the key information table of this formula, in order to provide reference for the development of reference samples and preparations of Yigongsan. Based on the results of the study, it is recommended that Panax ginseng should be removed the basal part of stem(rhizoma), Poria cocos should be removed the peel, Citrus reticulata should be cut into shreds and Glycyrrhiza uralensis should be used. According to 4.13 g/Qian(钱), 1 g/slice for ginger, 3 g for each jujube and 300 mL/Zhan(盏), the doses of Ginseng Radix, Poria, Atractylodis Macrocephalae Rhizoma, Glycyrrhizae Radix et Rhizoma, Citri Reticulatae Pericarpium, Zingiberis Rhizoma Recens, Jujubae Fructus were 1.652, 1.652, 1.652, 1.652, 1.652, 5, 6 g, and the total amount was 19.26 g. The decocting method was to crush the medicinal materials into fine powder with 50-80 mesh, add 300 mL of water and decoct to 210 mL for each dose, then remove the dregs and take it warmly. This formula was recorded in ancient books as the main treatment for the cold-deficiency of spleen and stomach, and Qi stagnation in children with vomiting and diarrhea and lack of appetite. It has been flexibly applied by later generations of physicians, and is often used to treat anorexia, inflammation of the digestive tract, diarrhea and other diseases in children.
10.Mechanism of Shugan Huazheng Prescription Against Liver Fibrosis Based on HIF-1α/VEGF/TGF-β1 Pathway
Anli XING ; Kunpeng ZHAO ; Qiuju ZHANG ; Jiena LI ; Shiyu CHEN ; Jiaqi GUO ; Ming ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(8):57-65
ObjectiveTo observe the therapeutic effect of Shugan Huazheng prescription on hepatic fibrosis model rats induced by carbon tetrachloride (CCl4) and explore whether it plays its role through hypoxia-induced factor-1α/vascular endothelial growth factor/transforming growth factor-β1 (HIF-1α/VEGF/TGF-β1) pathway. MethodA total of 54 male SPF SD rats were randomly divided into six groups: blank group, model group, colchicine group (0.2 mg·kg-1), and high-, medium-, and low-dose groups (29.52, 14.76, and 7.38 g·kg-1) of Shugan Huazheng prescription, with nine rats in each group. The molding was conducted three times a week for eight weeks. Administration began the day after the first injection, and the drug intervention was once a day for eight weeks. On the day after the last administration, the rats were deprived of food and water, and they were killed the next day, during which the physiological status of each group of rats was dynamically monitored. The pathological changes in the liver were observed by hematoxylin-eosin (HE) staining, and the content of hydroxyproline (HYP) and angiotensin Ⅱ (AngⅡ) in liver tissue were detected by enzyme-related immunosorbent assay (ELISA). Real-time fluorescent quantitative PCR (Real-time PCR) was used to determine the mRNA expression levels of HIF-1α, VEGF, and TGF-β1 in liver tissue, and immunohistochemical method (IHC) and Western blot were used to detect the protein expression levels of HIF-1α, VEGF, and TGF-β1 in liver tissue. ResultCompared with the blank group, the overall condition of rats in the model group decreased significantly. The proliferation of connective tissue and the increase in adipose cells between hepatocytes were obvious. The content of HYP and Ang was increased. The mRNA and protein expressions of HIF-1α, VEGF, and TGF-β1 were increased to varying degrees (P<0.05). Compared with the model group, the proliferation of connective tissue and inflammatory cell infiltration in the liver tissue of colchicine and Shugan Huazheng prescription groups were reduced. The content of HYP and Ang was decreased. The mRNA and protein expression levels of HIF-1α, VEGF, and TGF-β1 were decreased, and the colchicine group and high-dose group of Shugan Huazheng prescription were the most significant (P<0.05). ConclusionShugan Huazheng prescription has an obvious therapeutic effect on CCl4-induced hepatic fibrosis model rats. Its therapeutic mechanism may be related to the regulation of the HIF-1α/VEGF/TGF-β1 signaling pathway and the improvement of hepatic hypoxia, vascular remodeling, and the syndrome of Qi deficiency and blood stasis in hepatic fibrosis.


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