1.Action Mechanism of Huamoyan Granules in Treatment of Knee Osteoarthritis Based on TRPV1/p38 MAPK Pathway
Jin ZHANG ; Lili YANG ; Canwen ZHENG ; Jing KANG ; Yanlei MA ; Yue SHI ; Lei LI ; Hongxu MENG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(4):79-89
ObjectiveThis paper aims to observe the protective effect of Huamoyan granules on knee osteoarthritis (KOA) and explore whether its protective effect is oriented toward an anti-inflammatory direction by regulation of macrophage polarization, which can effectively inhibit the progression of pathological inflammatory response, reduce the release of inflammatory pain mediators, and downregulate the protein expression level of transient receptor potential vanilloid 1 (TRPV1), so as to provide experimental evidence for its clinical application and investigate its action mechanism. MethodsAfter adaptive feeding, Sprague-Dawley (SD) rats were randomly divided into six groups: sham group, model group, celecoxib group, and high, medium, and low-dose synovitis granule groups (9.6, 4.8, 2.4 g·kg-1). The administration dose of celecoxib capsules was 20 mg·kg-1. There were 10 rats in the sham group and 12 rats in the model group and each administration group. A KOA animal model was established by means of intra-articular injection of sodium iodoacetate into the knee joint. From the 10th day of the experiment, each administration group was given intragastric administration at a dose of 10 mL·kg-1 for 4 weeks. General conditions of rats in each group were assessed daily. The pressure pain threshold (PPT) to mechanical stimulation and joint diameter were recorded. X-ray examination was performed on the right knee joints of rats for imaging analysis. Enzyme linked immunosorbent assay (ELISA) was performed to detect the tumor necrosis factor-α (TNF-α), serum interleukin-1β (IL-1β), and other pro-inflammatory cytokines in rat serum samples, as well as the expression levels of neurogenic inflammatory mediators such as nerve growth factor (NGF) and calcitonin gene-related peptide (CGRP). Histopathological changes in the knee joint synovial tissues were examined by hematoxylineosin (HE) staining. Safranin O-fast green staining was performed to observe and evaluate the degree of knee cartilage lesions. Western blot was employed to quantitatively analyze TRPV1, p38 mitogen-activated protein kinase (p38 MAPK), and phosphorylated (p)-p38 MAPK in rat knee synovial tissues. Immunofluorescence (IF) was used to measure and assess M1/M2 macrophage polarization. ResultsCompared with those in the sham group, the circumference and joint diameter of the right knee were markedly enlarged in the model group (P<0.01), while PPTs of rats showed a significant reduction (P<0.01). The contents of IL-1β, TNF-α, CGRP, and NGF in rats' serum were significantly elevated (P<0.01), and the synovial Krenn score was increased (P<0.01). The Mankin score of cartilage tissue was increased (P<0.01), and the protein expressions of TRPV1 and p-p38 MAPK/p38 MAPK were significantly upregulated (P<0.01). The experimental intervention significantly reduced the proportion of pro-inflammatory M1 macrophages in the total macrophage population (P<0.01), and the percentage of M2 macrophages was decreased (P<0.01). The M1/M2 macrophage ratio was significantly elevated (P<0.01). Knee joint diameters of all dose groups of Huamoyan granules and the celecoxib group were reduced (P<0.01) compared with those of the model group, and the PPT recovery speeds in the high and medium-dose groups of Huamoyan granules were more obvious (P<0.05). The contents of IL-1β, CGRP, and NGF in the rats' serum in all administration groups were significantly reduced (P<0.05, P<0.01), and the content of TNF-α in rats' serum was significantly reduced (P<0.01). All dose groups of Huamoyan granules demonstrated significant reductions in both synovial Krenn score (P<0.05, P<0.01) and protein expression of TRPV1 and p-p38 MAPK/p38 MAPK in rats' synovial tissues (P<0.01). The percentage of M1 macrophages in the synovial tissues of the celecoxib group and all dose groups of Huamoyan granules was decreased (P<0.01). The percentage of M2 macrophages was increased (P<0.05), and the M1/M2 ratio was decreased (P<0.01). ConclusionHuamoyan granules can alleviate the inflammatory response of KOA, reduce the release of inflammatory pain mediators, and downregulate TRPV1 protein expression by regulating macrophage polarization. Its mechanism may be related to the TRPV1/p38 MAPK signaling pathway, thereby achieving the effect of improving peripheral pain hypersensitivity in KOA.
2.Quality of leukoreduced pooled concentrated platelets prepared from whole blood under different storage temperatures and durations
Huijuan AN ; Hong SHAN ; Zheng LIU ; Jiaojiao ZHANG ; Jiaojie WANG ; Lili BIE ; Min LIU
Chinese Journal of Blood Transfusion 2026;39(5):603-609
Objective: To compare the in vitro quality differences of leukoreduced pooled concentrated platelets prepared from whole blood preserved at different temperatures and for various durations, determine the safe time window for refrigerated whole blood in platelet preparation, and provide experimental evidence for optimizing blood component preparation procedures and improving the comprehensive utilization rate of blood resources. Methods: A total of 324 units of 400 mL ACD-B anticoagulated whole blood were randomly divided into two groups and stored at 4℃ and 22℃, respectively. The buffy coat was separated at three time intervals: <6 h, 6-12 h, and >12 h (≤18 h) post-collection, and allowed to rest overnight at 22℃. On the following day, the buffy coats from each group were pooled to prepare leukoreduced pooled platelet concentrates (LPPCs). Cell counts were performed, and metabolic parameters including pH, glucose, and lactate levels were measured to evaluate metabolic status. Platelet in vitro function and activation were assessed by thromboelastography (TEG), platelet aggregation rate, and the expression of PAC-1 and CD62P. The differences between the two groups were compared. Results: For pooled concentrated platelets prepared from whole blood stored at 4℃ and 22℃ for <6 h and 6-12 h, there were no significant differences in platelet count, pH, glucose levels, lactic acid levels, thromboelastography (TEG), platelet aggregation rate, or platelet activation rate (P>0.05). With prolonged refrigeration time of whole blood, compared with pooled concentrated platelets prepared from whole blood stored at 22℃ for >12 h but ≤18 h, those prepared from whole blood stored at 4℃ for >12 h but ≤18 h showed a decreased platelet count (1 152.83±180.08 vs 1 368.83±134.86, P=0.040), a significantly increased ADP-induced aggregation rate (26.82±6.59 vs 13.88±10.21, P=0.030), and significantly elevated expression rates of PAC-1 and CD62P (72.64±6.74 vs 63.28±5.97, P=0.030). However, there were no significant differences in pH, glucose content, lactate content, or thromboelastography (P>0.05). Conclusion: There was no significant difference in the in vitro count, function, or activation of pooled concentrated platelets prepared from whole blood stored at 4℃ and 22℃ within 12 hours. However, statistically significant differences were observed between the mixed concentrated platelets prepared from whole blood stored at 4℃ and those stored at 22℃ for more than 12 hours but not exceeding 18 hours. These findings can provide a reference for the preparation methods and clinical application of refrigerated platelets.
3.Exploration of ethical issues in the multiple disciplinary team model for children with 46, XY disorders of sex development
Lili PAN ; Zhe SU ; Wei SU ; Ying ZU ; Chenxi LI ; Huiping SU ; Jingyu YOU ; Kexin JIN ; Rongfei ZHENG
Chinese Medical Ethics 2026;39(6):718-723
ObjectiveTo explore the ethical issues and coping strategies in the diagnosis and treatment of 46, XY disorders of sex development (46, XY DSD) under a multidisciplinary team (MDT) model, and to deepen the understanding of relevant ethical issues while accumulating experience. MethodsA retrospective collection was conducted on the data of children with 46, XY disorders of sex development under a single-center MDT model from January 2017 to December 2024, and the ethical issues involved in the diagnosis and treatment process were analyzed. ResultsA total of 24 MDT discussion documents involving 20 children were collected, including 17 with female social gender and 3 with male social gender. The age ranged from 0.5 to 15.8 years, with children under 8 years accounting for 75%. The ethical issues primarily involved how to ensure adequate informed consent, whether surgery was necessary and its timing, and whether gender selection is reasonable and its timing. Among the 10 children raised as females who completed the gender role psychological scale assessment, 5 demonstrated masculine tendencies, all of whom had pathogenic genetic variants. Furthermore, only 1 patient changed the social gender to male. ConclusionThe diagnosis and treatment of 46, XY DSD is relatively complex, requiring multidisciplinary collaboration. This process involves numerous ethical issues and necessitates adhering to medical ethical principles. Ensuring adequate informed consent is crucial, especially for younger children. The surgery and its timing selection must adhere to the principles of beneficence, non-maleficence, and leaving room for the future. Gender selection constitutes a focus and challenge in diagnosis and treatment. Psychological scales and genetic etiologies are conducive to judgments regarding gender selection, yet multiple factors in the real world may interfere with the gender selection.
4.Preliminary report of a fully automatic feedback perimeter based on eye gaze tracking technique in the detection of college students
Lingxiao ZHOU ; Lili ZHAO ; Xiujuan ZHENG
International Eye Science 2025;25(2):319-322
AIM: To observe the preliminary application of fully automatic feedback perimeter based on the eye gaze tracking technique in the college students and to verify its test performance.METHODS: Home computer, monitor and eye tracker were used to complete the new automatic feedback perimeter meter. Based on the two characteristic eye movements of fixation and pulsation, the abnormal visual field is determined by analyzing whether the participant perceives the visual target, line-of-sight scan path and the fixation towards the visual target. RESULTS: A total of 63 participants(119 eyes)were collected with valid eye movement data. The average time for all participants was 82.46±14.68 s, the average time for right eyes was 88.21±15.30 s, and average time for the left eyes was 76.42±11.29 s(P<0.05).CONCLUSION: The fully automatic feedback perimeter based on eye gaze tracking technique can realize automatic human-computer interaction, and the detection method is simple and easy, which shortens the time of perimetry and improves the experience of participants.
5.Effect of Mori Folium-Ginseng Radix et Rhizoma on Glucose and Lipid Metabolism and Mechanism in Mouse Model of Type 2 Diabetes Mellitus
Congyi LIU ; Ning WANG ; Jingjing XU ; Tingting WANG ; Na ZHENG ; Zimeng HUANG ; Lingling QIN ; Lili WU ; Tonghua LIU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(17):20-28
ObjectiveTo study the effect of the herb pair Mori Folium-Ginseng Radix et Rhizoma (HMG) on glucose and lipid metabolism in the mouse model of type 2 diabetes mellitus and decipher the possible treatment mechanism. MethodsThe db/db mice were chosen as the mouse model of type 2 diabetes mellitus and then treated with HMG at low and high doses (1.56, 3.12 g∙kg-1, respectively) or metformin (0.26 g∙kg-1) by gavage for 6 weeks. The normal group and the model group were treated with double distilled water at the same time according to body weight. The 8-h fasting blood glucose and body weight were measured once a week. The oral glucose tolerance test (OGTT) was conducted at the 6th week of dosing. The mice were sacrificed after the end of dosing. Serum levels of lipids [total cholesterol (TC), triglycerides (TG), high-density lipoprotein cholesterol (HDL), and low-density lipoprotein cholesterol (LDL)], liver function indicators [aspartate aminotransferase (AST) and alanine aminotransferase (ALT)], non-esterified fatty acids (NEFA), glycosylated serum protein (GSP), serum glucose (GLU), fasting insulin (FINS), and renal function indicators [creatinine (Crea) and blood urea nitrogen (BUN)] were measured by enzyme-linked immunosorbent assay. The protein levels of peroxidase proliferator-activating receptor gamma (PPARγ), acetyl coenzyme A carboxylase (ACC), and sterol regulatory element-binding protein-1 (SREBP-1) were determined by Western blot. The pathological changes in the liver and pancreas were examined. ResultsCompared with the normal group, the model group presented increased body weight, elevated levels of blood glucose, TG, TC, AST, ALT, GLU, NEFA, GSP, and HDL-C, up-regulated protein levels of ACC and SREBP-1, and down-regulated protein level of PPARγ (P<0.01). Meanwhile, the model group presented a large amount of lipid droplets and steatosis in the liver, as well as karyopyknosis and lymphocyte infiltration in the pancreas. Compared with the model group, the high- and low-dose HMG groups showed decreased body weight, declined levels of blood glucose, TG, TC, AST, ALT, GLU, NEFA, and GSP, and elevate level of HDL-C (P<0.05, P<0.01). Moreover, the two groups showcased reduced lipid droplets and steatosis in the liver, as well as enlarged islets with clear boundaries and alleviated lymphocyte infiltration and karyopyknosis. Western blot results showed that the high-dose herb pair group demonstrated down-regulated protein levels of ACC and SREBP-1 and up-regulated protein level of PPARγ (P<0.01). ConclusionThe HMG can effectively improve the glucose and lipid metabolism in db/db mice by regulating the expression of PPARγ, SREBP-1, and ACC.
6.Effect of Mori Folium-Ginseng Radix et Rhizoma on Glucose and Lipid Metabolism and Mechanism in Mouse Model of Type 2 Diabetes Mellitus
Congyi LIU ; Ning WANG ; Jingjing XU ; Tingting WANG ; Na ZHENG ; Zimeng HUANG ; Lingling QIN ; Lili WU ; Tonghua LIU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(17):20-28
ObjectiveTo study the effect of the herb pair Mori Folium-Ginseng Radix et Rhizoma (HMG) on glucose and lipid metabolism in the mouse model of type 2 diabetes mellitus and decipher the possible treatment mechanism. MethodsThe db/db mice were chosen as the mouse model of type 2 diabetes mellitus and then treated with HMG at low and high doses (1.56, 3.12 g∙kg-1, respectively) or metformin (0.26 g∙kg-1) by gavage for 6 weeks. The normal group and the model group were treated with double distilled water at the same time according to body weight. The 8-h fasting blood glucose and body weight were measured once a week. The oral glucose tolerance test (OGTT) was conducted at the 6th week of dosing. The mice were sacrificed after the end of dosing. Serum levels of lipids [total cholesterol (TC), triglycerides (TG), high-density lipoprotein cholesterol (HDL), and low-density lipoprotein cholesterol (LDL)], liver function indicators [aspartate aminotransferase (AST) and alanine aminotransferase (ALT)], non-esterified fatty acids (NEFA), glycosylated serum protein (GSP), serum glucose (GLU), fasting insulin (FINS), and renal function indicators [creatinine (Crea) and blood urea nitrogen (BUN)] were measured by enzyme-linked immunosorbent assay. The protein levels of peroxidase proliferator-activating receptor gamma (PPARγ), acetyl coenzyme A carboxylase (ACC), and sterol regulatory element-binding protein-1 (SREBP-1) were determined by Western blot. The pathological changes in the liver and pancreas were examined. ResultsCompared with the normal group, the model group presented increased body weight, elevated levels of blood glucose, TG, TC, AST, ALT, GLU, NEFA, GSP, and HDL-C, up-regulated protein levels of ACC and SREBP-1, and down-regulated protein level of PPARγ (P<0.01). Meanwhile, the model group presented a large amount of lipid droplets and steatosis in the liver, as well as karyopyknosis and lymphocyte infiltration in the pancreas. Compared with the model group, the high- and low-dose HMG groups showed decreased body weight, declined levels of blood glucose, TG, TC, AST, ALT, GLU, NEFA, and GSP, and elevate level of HDL-C (P<0.05, P<0.01). Moreover, the two groups showcased reduced lipid droplets and steatosis in the liver, as well as enlarged islets with clear boundaries and alleviated lymphocyte infiltration and karyopyknosis. Western blot results showed that the high-dose herb pair group demonstrated down-regulated protein levels of ACC and SREBP-1 and up-regulated protein level of PPARγ (P<0.01). ConclusionThe HMG can effectively improve the glucose and lipid metabolism in db/db mice by regulating the expression of PPARγ, SREBP-1, and ACC.
7.Scientific Research Paradigm and Technological Layout Ideas Based on Clinical Dominant Disease of Traditional Chinese Medicine (Allergic Rhinitis)
Lili LIU ; Zheng GONG ; Linjing ZHANG ; Haiyang ZHANG ; Xiaoxiao ZHANG ; Zhanfeng YAN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(20):241-251
To implement the instructions and directives of General Secretary Xi Jinping on important documents related to traditional Chinese medicine (TCM), adhere to the equal emphasis on TCM and Western medicine, and promote mutual complementarity, win-win cooperation, and coordinated development between TCM and Western medicine, the China Association of Chinese Medicine has organized seminars on the clinical dominant diseases of TCM in multiple professional fields, achieving a series of results. The aim is to implement the research on dominant diseases of TCM described in the Law of the People's Republic of China on Traditional Chinese Medicine and the Opinions of the Central Committee of the Communist Party of China and the State Council on Promoting the Inheritance, Innovation and Development of Traditional Chinese Medicine, thus serving the construction of advantageous specialties, clinical talent cultivation, national scientific and technological layout, and academic innovation leadership. In the field of otolaryngology, the China Association of Chinese Medicine and many Chinese and Western medicine experts across the country have conducted multiple in-depth discussions and research. At the 10th Clinical Dominant Disease Series Youth Salon, they have elaborated and demonstrated the basic and specific suggestions and consensus on the clinical advantages of TCM and integrated TCM and Western medicine in the treatment of allergic rhinitis (AR), making great progress. However, there is still a lack of detailed research paths. Under the guidance of the China Association of Chinese Medicine and based on the TCM Dominant Disease Series Salon for AR, this study analyzed the difficulties encountered in AR diagnosis and treatment according to the occurrence and development law of AR. Based on the advantages and characteristics of AR diagnosis and treatment by TCM and integrated TCM and Western medicine, it explored the research paradigm and technological layout points of AR and put forward suggestions. The layout involved four aspects: Optimization of the AR diagnosis and treatment system, prevention and control of comorbidities, management of chronic diseases, and a platform for TCM inheritance. Additionally, suggestions for layout and research directions, expected goals and values, and priority levels for funding were proposed. The study is expected to provide a theoretical basis and development ideas for the future prevention and treatment of AR with TCM and integrated TCM and Western medicine and promote the high-quality development of TCM.
8.Investigation of the current status of radiation diagnosis and treatment resources and their use in Taiyuan City, China
Jianhua SHI ; Wenyan GUO ; Qiyu ZHAO ; Lixian ZHENG ; Li XU ; Fengjiao WU ; Xiaoai ZHAO ; Lili GUO
Chinese Journal of Radiological Health 2025;34(4):508-514
Objective To understand the current status of radiation diagnosis and treatment resources and their use in Taiyuan City, China, and provide data support for optimizing resource allocation and standardizing diagnosis and treatment. Methods A census-based approach was implemented using a standardized questionnaire to collect basic information on radiation diagnosis and treatment institutions in Taiyuan City. The number and use frequency of radiation diagnosis and treatment resources were calculated based on the resident population of Taiyuan City at the end of 2023. Results There were a total of 562 radiation diagnosis and treatment institutions in Taiyuan City, with
9.Intravitreal injection of Conbercept for the treatment of diabetic neovascular glaucoma
Lili CHEN ; Huiqin WU ; Bo ZHENG
International Eye Science 2025;25(11):1848-1853
AIM: To observe the efficacy of intravitreal Conbercept injection(IVC)in diabetic neovascular glaucoma(NVG).METHODS:Retrospective study. Totally 57 patients(57 eyes)with NVG(stage II)secondary to proliferative diabetic retinopathy(PDR)treated in our hospital were collceted from January 2019 to December 2022. Among them, 30 patients(30 eyes)were treated by IVC and panretinal photocoagulation(PRP)in the observation group, and 27 patients(27 eyes)were treated by PRP in the control group. The best corrected visual acuity(BCVA, LogMAR), intraocular pressure, neovascularization of the iridocorneal angle(NVA), central macular thinkness(CMT)changes and disease progression were analyzed with a follow-up of 2 years.RESULTS: There were no significant differences in baseline characteristics between the two groups(P>0.05). At the last follow-up, the BCVA in the observation group was better than the control group(P<0.05). The intraocular pressure, CMT and progression of NVA in the observation group were less than the control group(all P<0.05). The patients with NVG developed from stage II to stage III in the observation group was less than the control group(P<0.05). In the observation group, the BCVA improved compared with preoperative at 6 mo(P<0.05). In the control group, the BCVA decreased compared with preoperative after 24 mo(P<0.05). Multiple linear regression analysis indicated that NVG progression and intraocular pressure were factors affecting BCVA(LogMAR)changes(β=0.511, 0.014; t=4.461, 2.092; P<0.01, 0.041, respectively). In the observation group, the IOP decreased at 1, 6 and 12 mo compared with preoperative(all P<0.05), while there were no significant differences at 18 and 24 mo compared with preoperative(all P>0.05). In the control group, the intraocular pressure decreased at 6 mo compared with preoperative(P<0.05), while there were no significant differences at 1, 12, 18 and 24 mo compared with preoperative(all P>0.05). There were no significant difference about the types of glaucoma drugs between the observation group and the control group(all P>0.05). There were no infectious endophthalmitis and other complications observed in both groups.CONCLUSION: IVC combined with PRP demonstrates superior efficacy over PRP monotherapy in treating NVG(stage II)secondary to PDR. IVC combined with PRP showed better outcomes in controlling iris neovascularization, slowing visual impairment progression, reducing intraocular pressure, and delaying disease advancement.
10.Effect of Serum Containing Zhenwutang on Apoptosis of Myocardial Mast Cells and Mitochondrial Autophagy
Wei TANG ; Meiqun ZHENG ; Xiaolin WANG ; Zhiyong CHEN ; Chi CHE ; Zongqiong LU ; Jiashuai GUO ; Xiaomei ZOU ; Lili XU ; Lin LI
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(3):11-21
ObjectiveTo explore the effect of serum containing Zhenwutang on myocardial mast cell apoptosis induced by angiotensin Ⅱ (AngⅡ) and the mechanism of the correlation between apoptosis and mitochondrial autophagy. MethodsIn this experiment, AngⅡ and serum containing Zhenwutang with different concentrations were used to interfere with H9C2 cardiomyocytes for 24 h, and the survival rate of H9C2 cardiomyocytes was detected by cell counting kit-8 (CCK-8) to screen the optimal concentration for the experiment. Enzyme-linked immunosorbent assay (ELISA) was used to detect the content of B-type natriuretic peptide (BNP) in cell culture supernatant, and immunofluorescence was used to detect the cell surface area to verify the construction of the myocardial mast cell model. Subsequently, the experiment was divided into a blank group (20% blank serum), a model group (20% blank serum + 5×10-5 mol·L-1 AngⅡ), low-, medium-, and high-dose (5%, 10% and 20%) serum containing Zhenwutang groups, an autophagy inhibitor group (1×10-4 mol·L-1 3-MA), and autophagy inducer group (1×10-7 mol·L-1 rapamycin). The apoptosis level of H9C2 cells and the changes of mitochondrial membrane potential were detected by flow cytometry. The lysosomal probe (Lyso Tracker) and mitochondrial probe (Mito Tracker) co-localization was employed to detect autophagy. Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was used to detect Caspase-3, Caspase-9, B-cell lymphoma 2 (Bcl-2), Bcl-2-related X protein (Bax), and cytochrome C (Cyt C) in apoptosis-related pathways and the relative mRNA expression of ubiquitin ligase (Parkin), phosphatase and tensin homolog (PTEN)-induced kinase 1 (PINK1), and p62 protein in mitochondrial autophagy-related pathways. Western blot was used to detect cleaved Caspase-3, cleaved Caspase-9, Bax, Bcl-2, and Cyt C in apoptosis-related pathways, phosphorylated ubiquitin ligase (p-Parkin), phosphorylated PTEN-induced kinase 1 (p-PINK1), p62, and Bcl-2 homology domain protein Beclin1 in mitochondrial autophagy-related pathways, and the change of microtubule-associated protein 1 light chain 3 (LC3) Ⅱ/Ⅰ ratio. ResultsCCK-8 showed that when the concentration of AngⅡ was 5×10-5 mol·L-1, the cell activity was the lowest, and there was no cytotoxicity. At this concentration, the surface area of cardiomyocytes was significantly increased (P<0.01), and the content of BNP in the supernatant of culture medium was significantly increased (P<0.05). Therefore, AngⅡ with a concentration of 5×10-5 mol·L-1 was selected for the subsequent modeling of myocardial mast cells. Compared with the blank group, the model group and the autophagy inhibitor 3-MA group had a significantly increased apoptosis rate (P<0.01) and significantly decreased mitochondrial membrane potential (P<0.01). The results of immunofluorescence co-localization showed that compared with the blank group, the model group had a significantly decreased number of red and green fluorescence spots. The results of Real-time PCR showed that compared with that in the blank group, the relative mRNA expression of Bax, Caspase-3, Caspase-9, Cyt C, and p62 in the model group was significantly up-regulated (P<0.01), while the relative mRNA expression of Bcl-2, Parkin, and PINK1 was significantly down-regulated (P<0.01). In addition, the relative protein expression of Bax, cleaved Caspase-3, cleaved Caspase-9, Cyt C, and p62 was significantly up-regulated (P<0.01). The LC3Ⅱ/Ⅰ was significantly decreased, and the relative protein expression of Bcl-2, p-Parkin, p-PINK1, and Beclin1 was significantly down-regulated (P<0.01). Compared with the model group, the serum containing Zhenwutang groups and the autophagy inducer group had significantly decreased apoptosis rate (P<0.01), and the decrease ratio of mitochondrial membrane potential is significantly lowered (P<0.01) in a dose-dependent manner. Additionally, both red and green fluorescence spots became more in these groups. In the 3-MA group, the number of red and green fluorescence spots decreased significantly. The relative mRNA expression of Bax, Caspase-3, Caspase-9, Cyt C, and p62 was significantly down-regulated (P<0.05, P<0.01), while that of Bcl-2, Parkin, and PINK1 was significantly up-regulated (P<0.01). In the serum containing Zhenwutang groups, the relative protein expression levels of Bax, cleaved Caspase-3, cleaved Caspase-9, Cyt C, and p62 were significantly down-regulated (P<0.05,P<0.01). The LC3Ⅱ/Ⅰ was significantly increased, and the relative protein expression levels of Bcl-2, p-Parkin, p-PINK1, and Beclin1 were significantly up-regulated (P<0.01). ConclusionThe serum containing Zhenwutang can reduce the apoptosis of myocardial mast cells and increase mitochondrial autophagy. This is related to the inhibition of intracellular Bax/Bcl-2/Caspase-3 apoptosis pathway and regulation of Parkin/PINK1 mitochondrial autophagy pathway.

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