1.Proteomic Analysis of Danlou Tablet in Improving Platelet Function for Treating Coronary Heart Disease with Phlegm-stasis Intermingling Syndrome in Minipigs
Ziyan WANG ; Ying LI ; Aoao WANG ; Hongxu MENG ; Yue SHI ; Yanlei MA ; Guoyuan ZHANG ; Lei LI ; Jianxun LIU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(5):41-53
ObjectiveThis paper aims to observe the role of Danlou tablet in treating coronary heart disease (CHD) with phlegm-stasis intermingling syndrome in minipigs by improving platelet function and explore the potential pharmacological mechanism of Danlou tablet in regulating platelet function by using proteomics technology. MethodsThirty Bama minipigs were randomly divided into a normal control group (6 pigs) and a high-fat diet group (24 pigs). After 2 weeks of high-fat diet feeding, the high-fat diet group was randomly subdivided into a model group, an atorvastatin group (1 mg·kg-1), and Danlou tablet groups (0.6 g·kg-1 and 0.3 g·kg-1). All groups continued to receive a high-fat diet for 8 weeks after the procedure. The normal control group was given a regular diet, underwent only coronary angiography, and did not receive an interventional injury procedure. The model group and each administration group were fed a high-fat diet. Two weeks later, they underwent a coronary angiography injury procedure. After the procedure, drugs were mixed into the feed every morning for 8 consecutive weeks, with the minipigs maintained on a continuous high-fat diet during this period. Quantitative proteomics technology was further used to study platelet proteins, and differential proteins were obtained by screening. Bioinformatics analysis was performed to analyze key regulatory proteins and biological pathways involved in the therapeutic effect of Danlou tablet on CHD with phlegm-stasis intermingling syndrome. ResultsCompared with the normal control group, the model group showed a significant increase in total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C) of minipigs' serum (P<0.01), a significant shortening in prothrombin time of (PT) (P<0.01), a coagulation function index, and an increase in whole blood viscosity (P<0.01) and platelet aggregation rate (P<0.01). Moreover, the platelet morphology was altered, and the contents of endothelin-1 (ET-1) and nitric oxide (NO) were significantly increased (P<0.01). Hemodynamic parameters were obviously abnormal, including significantly decreased systolic blood pressure (SBP), diastolic blood pressure (DBP), mean arterial pressure (MAP), left ventricular systolic pressure (LVSP), and left ventricular maximal positive dp/dt (LV+dp/dtmax) (P<0.01). Left ventricular maximal negative dp/dt (LV-dp/dtmax) was significantly increased (P<0.01). Besides, there were myocardial cell hypertrophy, obvious edematous degeneration, massive interstitial inflammatory cell infiltration, high degree of fibrosis, and coronary endothelial atherosclerosis. TC and TG levels in minipigs' serum were significantly reduced in Danlou tablet groups with 0.6 g·kg-1 and 0.3 g·kg-1 (P<0.05, P<0.01), compared with those in the model group. LDL-C was decreased in the Danlou tablet group with 0.6 g·kg-1 (P<0.05). The whole blood viscosity under low and high shear conditions was significantly reduced in the Danlou tablet group with 0.6 g·kg-1 (P<0.05). In groups with all doses of Danlou tablet, maximum aggregation rate (MAR) and average aggregation rate (AAR) were significantly decreased (P<0.05, P<0.01), and platelets' morphological changes such as pseudopodia extension were reduced. ET-1 levels in the serum were significantly reduced. In the Danlou tablet group with 0.6 g·kg-1, NO level in the serum was reduced (P<0.05). In groups with all doses of Danlou tablet, DBP and MAP were significantly increased (P<0.05). In the Danlou tablet group with 0.6 g·kg-1, LVSP and LV+dp/dtmax were significantly increased (P<0.05, P<0.01), and LV-dp/dtmax was significantly decreased (P<0.05). In groups with all doses of Danlou tablet, edematous degeneration in myocardial tissue was milder, and coronary artery lesion degree was significantly alleviated. Compared with the normal control group, there were 94 differentially expressed proteins in the model group, including 81 up-regulated and 13 down-regulated proteins. Compared with the model group, the Danlou tablet group with 0.6 g·kg-1 showed 174 differentially expressed proteins, including 100 up-regulated and 74 down-regulated proteins. A total of 30 proteins were reversed after Danlou tablet intervention. Bioinformatics analysis revealed that its pharmacological mechanism may exert anti-platelet activation, aggregation, and adhesion effects through biological pathways such as regulation of actin cytoskeleton, platelet activation pathway, Fcγ receptor-mediated phagocytosis, as well as proteins such as growth factor receptor-bound protein 2 (GRB2), Ras-related C3 botulinum toxin substrate 2 (RAC2), RAC1, and heat shock protein 90 alpha family class A member 1 (HSP90AA1). ConclusionDanlou tablet can effectively reduce platelet activation and aggregation, exerting a good therapeutic effect on CHD with phlegm-stasis intermingling syndrome in minipigs. Its pharmacological mechanism may involve regulating biological pathways such as actin cytoskeleton and platelet activation pathway, as well as proteins like GRB2, RAC2, RAC1, and HSP90AA1, thereby exerting a pharmacological effect in anti-platelet activation, aggregation, and adhesion.
2.Epidemiological characteristics and disease burden of liver cancer in Guangdong Province
Ying ZHANG ; Yixuan CHEN ; Rong CAO ; Yue GAO ; Yutong HAN ; Ye WANG ; Ruilin MENG ; Xueyan ZHENG ; Yu LIAO ; Zhuanping ZENG
Journal of Public Health and Preventive Medicine 2026;37(1):68-72
Objective To analyze the epidemiological characteristics and disease burden of liver cancer in Guangdong Province in 2020, and to provide a scientific foundation for the development of regionalized prevention and control strategies for liver cancer. Methods According to the cancer registry data of Guangdong Province, the incidence, mortality and age-standardized rate by Chinese standard population in 2020 were calculated to analyze the epidemiological characteristics of liver cancer. The disability adjusted life years (DALYs), year of life loss (YLL), year of lived with disability (YLD), and cause-eliminated life expectancy were used to assess the disease burden of liver cancer. Results In 2020, the crude incidence rate and the age-standardized incidence rate of liver cancer in Guangdong Province were 27.79/100 000 and 20.84/100 000,respectively, and the crude mortality rate and the age-standardized mortality rate of liver cancer were 25.49/100,000 and 17.64/100 000, respectively. The total DALY and DALY rate of liver cancer in Guangdong Province were 515 311 person-years and 513.83/100 000, respectively. After eliminating the causes of death from liver cancer, the life expectancy in Guangdong Province increased from 84.60 years to 84.99 years. All indicators consistently demonstrated that the burden of liver cancer was higher in males than that in females, and the burden of liver cancer was higher in rural areas than that in urban areas. Conclusion Liver cancer in Guangdong Province exhibits a high incidence, mortality and disease burden level in 2020. There are obvious differences of gender, age and region in cancer burden. It is necessary to strengthen liver cancer screening and diagnosis and treatment in men, the elderly and those in rural areas to reduce the burden of liver cancer gradually in Guangdong Province.
3.Characteristics of 150 patients with spinal cord injury complicated with spasticity
Xiaolei LU ; Yiji WANG ; Genlin LIU ; Ying ZHENG ; Chunxia HAO ; Ying ZHANG ; Haiqiong KANG ; Bo WEI ; Qianru MENG ; Hongjun ZHOU
Chinese Journal of Rehabilitation Theory and Practice 2026;32(4):393-398
ObjectiveTo analyze the characteristics of 150 patients with spinal cord injury complicated with spasticity. MethodsA cross-sectional survey was conducted on 150 patients with spinal cord injury accompanied by spasticity from September, 2019 to December, 2024. Their age, gender, cause of injury, injury site, severity of injury, spasticity severity and other indicators were recorded. The relationships between different characteristics were analyzed, and a correlation analysis of disease duration, spasticity grade, injury level, injury severity and age were conducted. ResultsThere was no significant difference in age distribution between patients with tetraplegia and paraplegia (Z = 0.806, P = 0.420). The proportions of trauma (χ2 = 3.982, P = 0.046) and tetraplegia (χ2 = 10.559, P = 0.010) were higher in males than in females. Trauma was the main cause of injury in both tetraplegia and paraplegia patients; the proportion of tetraplegia was higher than paraplegia in trauma patients, while paraplegia was higher than tetraplegia in non-trauma patients (χ2 = 11.885, P < 0.001). Patients with tetraplegia was dominated by incomplete injury, whereas patients with paraplegia was dominated by complete injury (χ2 = 10.885, P = 0.012). Grade A injury was predominant in trauma patients (P = 0.003). Spasticity grade showed a very weak positive correlation with disease duration (r = 0.175, P = 0.032) and age (r = 0.168, P = 0.040). Injury severity showed a very weak positive correlation with age (r = 0.183, P = 0.025). ConclusionCharacteristics of patients with spinal cord injury complicated with spasticity is different with gender, cause of injury, injury level, injury severity.
4.Analysis of risk factors for cardiovascular events and construction of a nomogram prediction model in patients undergoing long-term peritoneal dialysis
Xinyuan ZHOU ; Yuxin JIANG ; Xiaoxia WANG ; Xiangjie YANG ; Runzhe ZHOU ; Yuqing MENG ; Dingxin ZHANG ; Jin ZHANG ; Ying WANG
Acta Universitatis Medicinalis Anhui 2026;61(4):748-757
ObjectiveTo analyze the risk factors for long-term cardiovascular events in patients undergoing long-term peritoneal dialysis (PD), and to construct and validate a visual nomogram prediction model based on multiple parameters. MethodsA prospective cohort study was conducted, consecutively enrolling 248 maintenance PD patients (dialysis duration ≥ 3 months). Demographic characteristics, clinical indicators, laboratory parameters, and echocardiographic indices (including left ventricular ejection fraction [LVEF], ratio of early diastolic mitral inflow velocity to early diastolic mitral annular velocity (E/e’), etc.) were collected. The composite endpoint was defined as the occurrence of cardiovascular events or cardiovascular death, with non-cardiovascular death as the competing risk and loss to follow-up or the end of follow-up as censoring events. Fine-Gray competing risks model was used to screen independent predictors, based on which a nomogram model was constructed. Internal validation was performed using the Bootstrap method (1 000 resamplings), and the concordance index (C-index) and time-dependent receiver operating characteristic (time-dependent ROC) curve were calculated to evaluate the model performance. ResultsWith a median follow-up of 29 months (interquartile range: 24–35 months), 88 patients (35.48%) reached the composite endpoint, including 80 cases of cardiovascular events and 8 cases of cardiovascular death, and 4 patients died of non-cardiovascular causes. Multivariate Fine-Gray analysis revealed that age, diabetes mellitus, hemoglobin (HGB) level and E/e' ratio were independent influencing factors of the composite endpoint. Specifically, each 1-year increase in age was associated with a 3.0% increase in the risk of the composite endpoint (HR=1.030, P=0.006); patients with diabetes mellitus had a 167.9% higher risk compared with non-diabetic patients (HR=2.679, P=0.007); each 1g/L increase in HGB level contributed to a 1.5% reduction in the risk (HR=0.985, P=0.003); and each 0.1 increase in E/e' ratio led to a 7.2% increase in the risk (HR=1.072, P=0.045). The nomogram model had a C-index of 0.76 (95% CI: 0.698–0.820), and the AUC of the time-dependent ROC curve reached 0.849 at 23 months of follow-up. ConclusionIncreased age, complicated with diabetes mellitus, decreased HGB, and elevated E/e' ratio are independent risk factors of long-term occurrence of cardiovascular events and cardiovascular death in patients undergoing long-term PD. The nomogram model constructed based on the above variables has good predictive value and clinical applicability, which can provide a reference for cardiovascular risk stratification and individualized intervention in long-term PD patients.
5.Research progress on strategies for toxicity reduction and efficacy enhancement of triptolide
Xiaoqing ZHENG ; Ying DING ; Shanshan XU ; Long WANG ; Shanshan HAN ; Yaping XING ; Meng ZHANG ; Wenhao LI
China Pharmacy 2026;37(11):1496-1501
Triptolide (TP), the core active component of the traditional Chinese medicine Tripterygium wilfordii , exhibits remarkable pharmacological activities including anti-inflammatory, immunosuppressive and anti-tumor effects, and holds broad application prospects in the treatment of major diseases such as autoimmune diseases and malignant tumors. However, TP has a narrow therapeutic window and causes multi-organ toxicities including liver, kidney and reproductive toxicities, which severely restrict its safe clinical application and new drug development. Therefore, toxicity reduction and efficacy enhancement has become a core scientific problem urgently to be solved in this field. This paper systematically reviews the four core strategies for TP toxicity reduction and efficacy enhancement, including structural modification, dosage form improvement, herbal compatibility, and external therapies of traditional Chinese medicine. Among them, structural modification optimizes the toxic and efficacy characteristics of TP from the molecular structure level, with typica l derivatives including (5 R )-5-hydroxy triptolide, ZT01, PG490-88, etc. Dosage form modification achieves toxicity reduction and efficacy enhancement via targeted and sustained-controlled drug release of diverse delivery systems. It includes triptolide preparations such as nanoparticles, liposomes, microemulsion gels and liquid crystals, possessing favorable clinical transformation potential. The herbal compatibility and external therapies of traditional Chinese medicine conform to the holistic view of traditional Chinese medicine and have a profound clinical application foundation, but their mechanisms of action are insufficiently elucidated, and they lack unified standardized specifications and high-quality evidence-based proof. In the future, we should rely on multi-omics technology to elucidate the toxic and efficacy mechanisms, integrate technologies to optimize preparations, improve the evaluation system and promote clinical transformation.
6.Analysis of Volatile Component Variations in Coptidis Rhizoma Processed with Euodiae Fructus Juice by HS-SPME-GC-MS Combined with Chemometrics and ROAV
Lin YAN ; Yun WANG ; Puling LI ; Lingbang MENG ; Zhe JIA ; Ying LI ; Ying LIU ; Min JIN ; Cun ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(21):218-225
ObjectiveTo analyze the volatile components in raw Coptidis Rhizoma, Euodiae Fructus juice-processed Coptidis Rhizoma (Yuhuanglian), and the processing adjuvant Euodiae Fructus juice, identify marker volatile components in the processed product, and elucidate the material basis changes during the processing with Euodiae Fructus juice. MethodsHeadspace solid-phase microextraction coupled with gas chromatography-quadrupole-time-of-flight mass spectrometry (HS-SPME-GC-Q-TOF-MS) was employed to analyze the volatile components in raw Coptidis Rhizoma, Yuhuanglian, and Euodiae Fructus juice. Chemometric analysis was performed to compare volatile components among samples and screen the marker components. Finally, the relative odor activity value (ROAV) was calculated to investigate odor characteristics. ResultsA total of 127 components were detected, with 27, 75, 84 volatile components identified in Euodiae Fructus juice, raw Coptidis Rhizoma, and Yuhuanglian, respectively. Processing with Euodiae Fructus juice significantly increased the number and content of volatile components. Chemometric analysis revealed significant changes in volatile components among the three sample types, allowing clear discrimination. After processing, 39 components such as methyl octanoate and allyl propionate were introduced, while 34 components including furfural and hexanoic acid increased in content, and 11 components including butyric acid and octanal decreased. Additionally, eight components, including zingiberene, 6-methyl-6-hepten-2-one, and 5-methyl-2-hexanone, were screened as marker components of Yuhuanglian. Moreover, processing enhanced the ROAV values of 2,5-dimethylpyrazine (roasted, nutty, coffee-like aroma), 3-ethylphenol, and guaiacol (smoky, woody aroma). Additionally, it incorporated 1-hepten-3-one from Euodiae Fructus juice and the newly generated 1-hexen-3-one, contributing mushroom and cucumber-like odors. The sensory profile of Yuhuanglian integrated nutty, roasted, smoky, and woody notes with the citrus and fruity aroma originally contributed by octanal. ConclusionThe processing with Euodiae Fructus juice introduced components from adjuvant, significantly altering the volatile component profile and enhancing the sensory odor characteristics of Yuhuanglian. This study provides a valuable reference for exploring the material basis of volatile components and establishing processing specifications for Yuhuanglian.
7.Analysis of Volatile Component Variations in Coptidis Rhizoma Processed with Euodiae Fructus Juice by HS-SPME-GC-MS Combined with Chemometrics and ROAV
Lin YAN ; Yun WANG ; Puling LI ; Lingbang MENG ; Zhe JIA ; Ying LI ; Ying LIU ; Min JIN ; Cun ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(21):218-225
ObjectiveTo analyze the volatile components in raw Coptidis Rhizoma, Euodiae Fructus juice-processed Coptidis Rhizoma (Yuhuanglian), and the processing adjuvant Euodiae Fructus juice, identify marker volatile components in the processed product, and elucidate the material basis changes during the processing with Euodiae Fructus juice. MethodsHeadspace solid-phase microextraction coupled with gas chromatography-quadrupole-time-of-flight mass spectrometry (HS-SPME-GC-Q-TOF-MS) was employed to analyze the volatile components in raw Coptidis Rhizoma, Yuhuanglian, and Euodiae Fructus juice. Chemometric analysis was performed to compare volatile components among samples and screen the marker components. Finally, the relative odor activity value (ROAV) was calculated to investigate odor characteristics. ResultsA total of 127 components were detected, with 27, 75, 84 volatile components identified in Euodiae Fructus juice, raw Coptidis Rhizoma, and Yuhuanglian, respectively. Processing with Euodiae Fructus juice significantly increased the number and content of volatile components. Chemometric analysis revealed significant changes in volatile components among the three sample types, allowing clear discrimination. After processing, 39 components such as methyl octanoate and allyl propionate were introduced, while 34 components including furfural and hexanoic acid increased in content, and 11 components including butyric acid and octanal decreased. Additionally, eight components, including zingiberene, 6-methyl-6-hepten-2-one, and 5-methyl-2-hexanone, were screened as marker components of Yuhuanglian. Moreover, processing enhanced the ROAV values of 2,5-dimethylpyrazine (roasted, nutty, coffee-like aroma), 3-ethylphenol, and guaiacol (smoky, woody aroma). Additionally, it incorporated 1-hepten-3-one from Euodiae Fructus juice and the newly generated 1-hexen-3-one, contributing mushroom and cucumber-like odors. The sensory profile of Yuhuanglian integrated nutty, roasted, smoky, and woody notes with the citrus and fruity aroma originally contributed by octanal. ConclusionThe processing with Euodiae Fructus juice introduced components from adjuvant, significantly altering the volatile component profile and enhancing the sensory odor characteristics of Yuhuanglian. This study provides a valuable reference for exploring the material basis of volatile components and establishing processing specifications for Yuhuanglian.
8.Visualization of Wallerian Degeneration of The Corticospinal Tract and Glial Pathological Changes After Ischemic Stroke in Mice
Meng MENG ; Jiao MU ; Mei-Yu LI ; Jun-Yan GAO ; Xiao-Min ZHU ; Liang-Liang LIU ; Ying GUO ; Guo-Hui ZHANG
Progress in Biochemistry and Biophysics 2026;53(10):2808-2819
ObjectiveStroke remains the leading cause of acquired disability in adults worldwide, imposing a substantial socioeconomic burden on patients, families, and healthcare systems. Among the various neurological deficits caused by stroke, motor dysfunction——particularly contralateral hemiplegia——is the most common and debilitating consequence, and it predominantly results from damage to the cortical motor conduction pathways. When motor cortex neurons or their projecting fibers are damaged, the distal axons and their associated myelin sheaths undergo a series of progressive, programmed degeneration, fragmentation, and eventual phagocytic clearance. This pathological process is termed Wallerian degeneration (WD). However, the temporal progression of WD following ischemic stroke and the associated alterations in the local glial microenvironment remain incompletely understood. The present study was designed to systematically elucidate the time course of corticospinal tract (CST) axonal WD after ischemic stroke in mice, and to simultaneously characterize the dynamic pathological changes of astrocytes, microglia, oligodendrocyte lineage cells, and myelinating oligodendrocytes during the WD process. MethodsEight-week-old male C57BL/6J mice were randomly assigned to control and ischemic stroke groups. A focal cortical ischemic stroke model was established in the right sensorimotor cortex using the photothrombotic method. This method induces localized intravascular thrombosis in the target cortical region through precisely controlled interaction between the photosensitizer and laser light of specific wavelength. At day 1 post‑surgery, 2,3,5-triphenyltetrazolium chloride (TTC) staining was performed to evaluate infarct volume, hematoxylin and eosin (HE) staining for histopathological examination, and Nissl staining for assessment of neuronal injury. For axonal degeneration time-course analysis, rAAV-CAG-mCherry anterograde viral tracing was employed to specifically label CST neurons. Cervical spinal cord sections at the CST region were collected at day 3, 5, 7, 14, and 60 post-injury. Morphological changes of mCherry-positive CST fibers were observed under fluorescence microscopy, and the fluorescence signal area fraction within the defined region was quantitatively analyzed to evaluate the dynamic process of axonal degeneration. For glial response assessment, immunofluorescence staining against Iba1 (microglia), GFAP (astrocytes), Olig2 (oligodendrocyte lineage cells), and APC (myelinating oligodendrocytes) was performed at day 7, 14, and 60 post‑surgery. The numbers of positive cells were counted to quantitatively analyze the dynamic changes in each glial population at different time points after ischemia, thereby revealing the evolution of the glial microenvironment during WD. ResultsThe photothrombotic method stably and reliably established an ischemic brain injury model confined to the sensorimotor cortex. Viral tracing revealed that the proportion of mCherry‑positive CST fibers was significantly reduced at day 7 after ischemic injury to the neuronal soma. Immunofluorescence staining demonstrated that during WD, the numbers of microglia and astrocytes in the CST region were significantly increased, whereas the numbers of oligodendrocyte lineage cells and myelinating oligodendrocytes showed no significant changes. ConclusionWD of CST axons is observed at day 7 post-ischemic stroke in a delayed manner. This degenerative process selectively activates microglia and astrocytes, while leaving the number of oligodendrocyte lineage cells largely unaffected.
9.Neonatal alloimmune thrombocytopenia complicated with piperacillin drug antibody: a case report
Zifan MENG ; Shaoqiang ZHANG ; Qiang JU ; Ying LI ; Songxia YAN ; Haiyan WANG
Chinese Journal of Blood Transfusion 2025;38(7):969-974
Objective: To investigate the clinical symptoms, laboratory tests, and treatment strategies of a case of fetal/neonatal alloimmune thrombocytopenia (FNAIT) complicated with piperacillin drug antibody. Methods: The platelet antibodies in the mother were screened and identified by ELISA. The HLA antigens of the newborn were genotyped through PCR-SSO, while the specificity of HLA antibodies in the mother was determined using a Single Antigen kit. The drug antibody was detected by a piperacillin kit. Results: Maternal antibodies against paternally-derived platelet antigens were detected. The HLA genotypes of the newborn were identified as HLA A
33∶03 and HLA B
58∶01. The mother exhibited strong positive antibodies against the specific platelet antigens of the newborn, namely anti-HLA-A33 and anti-HLA-B58 antibodies. The piperacillin antibody was detected in the newborn. Following treatment of continuous intravenous immunoglobulin (IVIG), platelet transfusions, red blood cell transfusions and discontinuation of piperacillin treatment, the platelet count and hemoglobin levels increased in the newborn. Conclusion: The newborn in this case was diagnosed with FNAIT complicated by the presence of anti-HLA-A33 and anti-HLA-B58 antibodies, as well as drug-induced hemolytic anemia caused by piperacillin drug antibody. The condition is more complicated under the influence of dual immune antibodies. Laboratory detection techniques such as platelet antibody and drug antibody tests can assist in early clinical diagnosis. At the same time, more active drug and blood transfusion treatments should be given in clinical practice to improve the prognosis.
10.Mechanism of tannins from Galla chinensis cream in promoting skin wound healing in rats based on FAK/PI3K/Akt/mTOR signaling pathway.
Wen YI ; Zi-Yi YAN ; Meng-Qiong SHI ; Ying ZHANG ; Jie LIU ; Qian YI ; Hai-Ming TANG ; Yi-Wen LIU
China Journal of Chinese Materia Medica 2025;50(2):480-497
This study investigated the effects and action mechanism of tannins from Galla chinensis cream(TGCC) on the skin wound of rat tail. Male Sprague Dawley(SD) rats were randomly divided into a control group, model group, model+low-dose TGCC(50 mg per rat) group, model+high-dose TGCC group(100 mg per rat), and model+TGC+FAK inhibitor(Y15) cream(100 mg+10 mg per rat) group, with 10 rats in each group. After the rat tail skin injury model was successfully constructed, in the treatment group, corresponding drugs were applied to the wound surface, while in the control and model groups, the same amount of cream base as the TGCC group was applied by the same method. Then, sterile gauze was wrapped around the wound edge, and these operations were performed three times a day for 28 consecutive days. The wound healing status at the third, seventh, eleventh, fourteenth, twenty-first, and twenty-eighth days was recorded, and the wound healing rate and healing time were calculated. On the day after the last dose of medication, rat serum and tail skin wound tissue were collected for analyzing the activities of serum alanine aminotransferase(ALT), aspartate aminotransferase(AST), creatinine(CREA), urea, reactive oxygen species(ROS), interferon gamma(IFN-γ), interleukin(IL)-1β, IL-6, IL-4, IL-10, tumor necrosis factor(TNF)-α, as well as catalase(CAT), glutathione(GSH), lactate dehydrogenase(LDH), malondialdehyde(MDA), myeloperoxidase(MPO), superoxide dismutase(SOD), total antioxidant capacity(T-AOC), platelet endothelial cell adhesion molecule-1(CD31), and leukocyte differentiation antigen 34(CD34) in the wound tissue of rat tail skin. Hematoxylin-eosin, Masson, and sirius red staining were used to observe the morphological changes in the wound tissue of rat tail skin. The thickness of the epidermis, the number of fibroblasts and blood vessels, and the contents of collagen fibers, typeⅠ collagen(COLⅠ), and COLⅢ were calculated. The mRNA expressions of keratin 10(KRT10), KRT14, vascular endothelial growth factor(VEGF), fibroblast growth factor(FGF), epidermal growth factor(EGF), CD31, CD34, matrix metallopeptidase-2(MMP-2), MMP-9, COLⅠ, COLⅢ, desmin, fibroblast specific protein 1(FSP1), IFN-γ, IL-1β, TNF-α, IL-4, IL-6, and IL-10 in skin wound tissue were determined by quantitative real-time polymerase chain reaction(PCR). Western blot was utilized to detect the protein expressions of KRT10, KRT14, VEGF, FGF, EGF, MMP-2, MMP-9, COLⅠ, COLⅢ, desmin, FSP1, focal adhesion kinase(FAK), phosphorylated focal adhesion kinase(p-FAK), phosphatidylin-ositol-3-kinase(PI3K), phosphorylated phosphatidylin-ositol-3-kinase(p-PI3K), protein kinase B(Akt), phosphorylated protein kinase B(p-Akt), mammalian target of rapamycin(mTOR), and phosphorylated mammalian target of rapamycin(p-mTOR). The results manifest that TGCC can dramatically elevate the healing rate of rat tail wounds and shorten wound healing time. Besides, it can reduce serum ROS levels, the contents of MDA, MPO, and LDH in the rat skin wound tissue, as well as the serum IFN-γ, IL-1β, IL-6, and TNF-α levels and the mRNA expression levels of IFN-γ, IL-1β, IL-6, and TNF-α in the skin wound tissue. It can elevate the activities of CAT, GSH, SOD, and T-AOC in wound tissue, the IL-4 and IL-10 contents in serum, and the mRNA expressions of IL-4 and IL-10 in the wound tissue. In addition, TGGC can inhibit inflammatory cell infiltration and increase the epidermal thickness, counts of fibroblasts and blood vessels, and contents of collagen fibers, COLⅠ, and COLⅢ. Besides, TGCC can elevate the mRNA and protein expressions of epidermal differentiation markers(KRT10 and KRT14), endothelial cell markers(CD31 and CD34), angiogenesis and fibroblast proliferation, differentiation markers(VEGF, FGF, EGF, COLⅠ, COLⅢ, desmin, and FSP1), reduce the mRNA and protein expressions of gelatinases(MMP-2 and MMP-9), and increase protein expressions of p-FAK, p-PI3K, p-Akt, p-mTOR, as well as ratios of p-FAK/FAK, p-PI3K/PI3K, p-Akt/Akt, and p-mTOR/mTOR. These results suggest that TGCC can significantly facilitate skin wound healing, and its mechanism may be related to the activation of the FAK/PI3K/Akt/mTOR signaling pathway, inhibition of inflammatory cell infiltration in skin wound tissue, elevation of epidermal thickness, counts of fibroblasts and vessels, and contents of collagen fiber, COLⅠ, and COLⅢ, and reduction of MMP-2 and MMP-9 expressions, thus accelerating wound healing.
Animals
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Male
;
Wound Healing/drug effects*
;
Rats
;
Rats, Sprague-Dawley
;
Signal Transduction/drug effects*
;
TOR Serine-Threonine Kinases/genetics*
;
Phosphatidylinositol 3-Kinases/genetics*
;
Skin/metabolism*
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Proto-Oncogene Proteins c-akt/genetics*
;
Tannins/pharmacology*
;
Humans
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Drugs, Chinese Herbal/administration & dosage*
;
Focal Adhesion Kinase 1/genetics*


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