1.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.
2.Effect of hypertension on cardiovascular fibrosis and sFRP2 expression in rats
Yao XU ; Chunhui MA ; Zhiyong LI
Journal of Pharmaceutical Practice and Service 2025;43(4):180-184
Objective To investigate the effect of spontaneous hypertension on the remodeling of cardiac and aortic tissues in rats, with special attention to the changes in the content of collagen fibers, elastic fibers and secreted Frizzled-related protein 2 (sFRP2) in cardiac and aortic tissues. Methods 28-week-old SHR rats (Spontaneously Hypertensive rats) and WKY (Wistar-Kyoto rats) of the same age were selected as experimental animals. Cardiac load was assessed by calculating the cardiac weight index. Collagen fibers and elastic fibers were isolated from the rat thoracic aorta by hot alkali method, and their content was determined by biochemical analysis. In addition, pathological evaluation of tissue sections of the left ventricle and thoracic aorta were performed by H&E staining, Sirius red staining, and lichen red staining. Western blotting was used to determine the expression level of sFRP2 protein in cardiac tissues. Results Compared with WKY rats, the heart weight index of SHR rats increased significantly (P<0.001), and the results of biochemical analysis and staining of pathological sections showed that the content of collagen fibers in the aorta in the SHR group was higher than that in the WKY group, while the content of elastic fibers was lower, but the difference did not have statistical significance. The content of collagen fibers in the heart of the SHR group was significantly higher than that in the WKY group (P<0.01). Western blotting showed that there was no significant difference in the expression level of sFRP2 protein in heart tissues between the two groups. Conclusion The remodeling of cardiac and aortic tissues in a rat model of spontaneous hypertension may involve complex molecular mechanisms, not just changes in the content of collagen fibers and elastic fibers. The detailed mechanism of the progression of spontaneous hypertension and target organs damage still need further investigation.
3.Analysis of the basic situation of radiological diagnosis and treatment resources in medical institutions in Hunan Province, China
Zhenwei CAO ; Zhiyong XU ; Zipo ZHAI ; Junzhe PENG ; Donghui CHEN ; Yunfeng NIE
Chinese Journal of Radiological Health 2025;34(4):500-507
Objective To obtain the data of radiological diagnosis and treatment resource distribution at medical institutions of different levels and in various cities, understand the status of resource allocation, provide policy-making basis and suggestions for optimizing the allocation of radiological diagnosis and treatment resources within the province, and offer data and references for related research. Methods A basic situation questionnaire survey was conducted on radiological diagnosis and treatment institutions in Hunan Province. Data were reviewed, analyzed, and statistically processed using Excel software to understand the allocation situation of radiological diagnosis and treatment resources in Hunan Province. Results As of 2022, there were
4.Health-related quality of life among elderly patients with metabolic syndrome
DENG Tianrui ; WANG Zhiyong ; YE Qing ; TANG Wei ; YANG Bin ; XU Fei
Journal of Preventive Medicine 2025;37(4):325-330
Objective:
To investigate the health-related quality of life and its influencing factors in elderly patients with metabolic syndrome (MS), so as to provide the evidence for improving health-related quality of life in older adults with chronic diseases.
Methods:
In 2021, elderly MS patients aged ≥60 years from four districts in Nanjing City were selected as the study subjects using a multi-stage random sampling method. Data on social demographic information, lifestyle, disease history and blood biochemical indicators were collected through questionnaire surveys, physical examination and laboratory tests. Health utility value and EuroQol Visual Analog Scale (EQ-VAS) score were assessed using the EuroQol 5-dimension 3-level questionnaire. Factors affecting health-related quality of life were identified with the Tobit regression model and multiple linear regression model.
Results:
A total of 3 378 elderly MS patients were included, with a median age of 67.00 (interquartile range, 7.00) years. There were 1 558 males (46.12%) and 1 820 females (53.88%). The median (interquartile range) of health utility value and EQ-VAS score were 1.00 (0.03) and 80.00 (15.00). Tobit regression and multiple linear regression analysis showed that gender (female, β=-0.034), education level (middle school, β=0.024; junior college and above, β=0.046), marital status (married, β=0.014), physical activity (sufficient, β=0.013), vegetable intake (meet standard, β=-0.009) and fruit intake (meet standard, β=0.016) were the influencing factors of health utility value. Residence (urban area, β=1.933) and alcohol consumption (yes, β=1.761) were influencing factors of EQ-VAS score. Age, cardiovascular and cerebrovascular diseases, malignant tumors and chronic respiratory diseases were the influencing factors of health utility value and EQ-VAS score.
Conclusion
Age, sex, marital status, residence, lifestyle and disease are mainly associatied with the health-related quality of life in elderly MS patients.
5.Fundamental and symptomatic causes of myopia in children and adolescents: optimization and improvement of a comprehensive system
Chinese Journal of School Health 2024;45(12):1677-1680
Abstract
Prevention and control of myopia requires comprehensive measures. Based on the system established earlier, the system is revised and upgraded. The original system emphasizes comprehensive measures and proposes to focus on both the symptoms and root causes. These measures are summarized into six aspects, three address the root cause measures including health education, glasses optics and visual environment, and three address symptoms measures including eye exercises, physical therapy and drugs. The paper is comprehensively supplemented and analyzes the primary and the secondary, and promotes physical therapy from treating the symptoms to treat both the symptoms and the root causes. These improvement measures will help promote the prevention and control of myopia in children and adolescents.
6.Effects of polysaccharide content and anti-inflammatory,hemostatic,and antioxidant activities before and after preparation of Mosla chinensis-Jiangxiangru
Qiuting MA ; Lei XU ; Yuchen ZHU ; Mengting XU ; Wenkai ZHANG ; Zhiyong LIU
Chinese Journal of Comparative Medicine 2024;34(2):79-88
Objective Explore changes in polysaccharides in Jiangxiangru before and after ginger juice preparation,and evaluate polysaccharide anti-inflammatory and antioxidant activities before and after processing.Methods The contents of Jiangxiangru polysaccharide(JXRPs)and Ginger juice processed of Jiangxiangru polysaccharide(JZJXRPs)before and after processing were determined by phenol-sulfuric acid method.We used the swelling model in rats and endotoxin(LPS)to establish the RAW264.7 mouse macrophage inflammation model.The optimal administration concentration was determined using a cell proliferation(MTT)assay.Enzyme-linked immunosorbent assay(ELISA)were used to measure Interleukin-6(IL-6),Interleukin-12(IL-12),Nitric oxide(NO),Interleukin-4(IL-4),and Interleukin-10(IL-10).Bleeding time of mice by tail cutting was observed to evaluate the hemostatic effect.The ability to remove 1,1-diphenyl-2-picryl-hydrazyl radical(DPPH)and 2,2'-Azinobis-(3-ethylbenzthiazoline-6-sulphonate)(ABTS)was used to evaluate in vitro antioxidant activity.Results The contents of JXRPs and JZJXRPs were 13%and 22%,respectively.In swollen rats at 4 h after injection,compared with the model group,200 mg/kg JXRPs and 100 mg/kg JZJXRPs significantly reduced rat swelling(P<0.05).In vitro anti-inflammation assessment showed that the polysaccharides before and after processing significantly inhibited secretion of IL-6,IL-12,and NO(P<0.01)and promoted secretion of IL-4 and IL-10(P<0.01),and also that processing post-effects were stronger.The hemostatic experiment show that,compared with the control group,JXRPs increased hemostasis,but without a significant difference,and no significant difference was found using JZJXRPs,although high doses showed a trend to increase hemostasis.In vitro antioxidant activity showed that JXRPs and JZJXRPs had different scavenging abilities for DPPH and ABTS with IC50 values of JXRPs of 0.2215 and 0.2110 mg/ml,respectively,and IC50 values of JZJXRPs of 0.1651 and 0.1884 mg/mL,respectively.Conclusions After Jiangxiangru is produced in ginger juice,it promotes dissolution of polysaccharides and increases polysaccharide content.Anti-inflammatory,hemostasis,and antioxidant capacities are stronger in JZJXRPS than JXRPS,which lays the foundation for follow-up research and clinical applications of JXRPS.
7.In Vitro and in Vivo Evaluation of Scutellarin-phospholipid Complex Nanoemulsion and Analysis of Its Activity in Ameliorating LPS-induced Vascular Endothelial Injury
Tian LUO ; Zhiyong HE ; Xiangjun MAO ; Xue LIU ; Jinggang HE ; Yuan ZHI ; Xiangchun SHEN ; Qianli XU ; Ling TAO
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(10):159-168
ObjectiveTo evaluate some properties of scutellarin-phospholipid complex nanoemulsion(SCU-PC-NE), such as release, cell uptake and tissue distribution, and to investigate its effect on ameliorating lipopolysaccharide(LPS)-induced vascular endothelial injury. MethodSCU-PC-NE was prepared by weighting SCU-PC, ethyl oleate, Kolliphor HS15, 1,2-propylene glycol(50, 400, 514.3, 85.7 mg), respectively. And the appearance of SCU-PC-NE was observed by transmission electron microscope, the average paticle size and Zeta potential were measured by nanopotential particle size analyzer. The cumulative release of SCU-PC-NE in vitro was measured by dynamic dialysis, thiazolyl blue(MTT) colorimetric assay was used to investigate the effect of SCU-PC-NE on the viability of human umbilical vein endothelial cells(HUVECs), the inverted fluorescence microscope and flow cytometry were used to investigate cell uptake of HUVECs by SCU-PC-NE in vitro using coumarin 6 as a fluorescent probe, the tissue distribution of DiR/SCU-PC-NE labeled by near infrared fluorescent dyes was obeserved by small animal in vivo imaging system. The inflammation injury model was established by co-incubation with LPS(1 mg·L-1) and HUVECs, the effect of SCU-PC-NE on the levels of interleukin(IL)-1β and IL-6 were determined by enzyme-linked immunosorbent assay(ELISA), 18 Kunming male mice were randomly divided into blank group, model group, blank preparation group(equivalent to high dose group), SCU group and SCU-PC-NE low and high dose groups(5, 10 mg·kg-1), 3 mice in each group, and the drug administration groups were administered once in the tail vein at the corresponding dose every 48 h, equal volume of normal saline was given to the blank group and the model group, and the drug was administered for 4 consecutive times. Except for the blank group, the endothelial inflammatory injury was induced by intraperitoneal injection of LPS(10 mg·kg-1) at 12 h before the last administration in each group. Hematoxylin-eosin(HE) staining was used to investigate the effect of SCU-PC-NE on the histopathological changes in the thoracic aorta of mice. ResultThe appearance of SCU-PC-NE displayed pale yellow milky light, mostly spherical with rounded appearance and relatively uniform particle size distribution, with the average particle size of 35.31 nm, Zeta potential of 7.23 mV, and the encapsulation efficiency of 75.24%. The cumulative release in vitro showed that SCU-PC-NE exhibited sustained release properties compared with SCU. The cell viability of SCU-PC-NE was >90% at a concentration range of 1.05-8.4 mg·L-1. The results of cellular uptake experiments showed that the cellular uptake ability of SCU-PC-NE was significantly enhanced when compared with the SCU group(P<0.01). Compared with normal mice, the results of tissue distribution showed that the fluorescence intensity of DiR/SCU-PC-NE was significantly enhanced in the spleen, kidney, brain and thoracic aorta of mice at different time points after intraperitoneal injection of LPS(P<0.05, P<0.01), especially in thoracic aorta. ELISA results showed that the levels of IL-1β and IL-6 in the model group were significantly increased when compared with the blank group(P<0.05, P<0.01), and compare with the model group, all administration groups significantly down-regulated IL-1β level, with the strongest effect in the SCU-PC-NE high-dose group(P<0.01), and all administration groups significantly down-regulated IL-6 level, with the strongest effect in the SCU-PC-NE low-dose group(P<0.05). Compare with the blank group, the results of HE staining showed that the endothelial cells were damaged, the elastic fibers were broken and arranged loosely in the model group, although similar vascular injury could be observed in the blank preparation group, SCU group and SCU-PC-NE low-dose group, the vascular endothelial damage was significantly reduced in the high-dose group of SCU-PC-NE, which had a better effect than that in the SCU group. ConclusionSCU-PC-NE can promote the uptake of drugs by endothelial cells and effectively enriched in the site of vascular endothelial injury caused by LPS, suggesting that it has a protective effect on vascular endothelial injury and is a good carrier of SCU.
8.The increasement of blood METRNL protein by insulin sensitizer rosiglitazone
Fei XU ; Jin CHEN ; Zhiyong LI
Journal of Pharmaceutical Practice and Service 2024;42(4):165-168
Objective To investigate the effect of insulin sensitizer rosiglitazone on blood METRNL levels. Methods After fed with high fat diet (HFD) for 3 months, obese mice were treated with rosiglitazone for 1 month. Glucose tolerance was tested with glucose tolerance test (GTT), and METRNL levels in blood were measured by ELISA. Real time fluorescence quantitative PCR was used to detect the expression of METRNL in various tissues such as muscle, liver, white fat, brown fat, brain, spleen and kidney, as well as the expression of mitochondrial proteins in brown adipose tissue. Results Glucose tolerance of animals fed a high-fat diet was improved in rosiglitazone group, and blood METRNL levels were also increased significantly in this group. Rosiglitazone treatment increased the expression of METRNL in brown fat and kidney tissue. There was no effect on METRNL expression in muscle, liver, white fat, brain and spleen. Rosiglitazone increased the expression of mitochondrial-associated proteins in brown adipose tissue. Conclusion The insulin sensitizer rosiglitazone might increase the serum METRNL level by increasing the METRNL expression in brown fat and kidney tissue, suggesting that METRNL may be involved in the therapeutic effect of rosiglitazone on diabetes.
9.Diagnosis and treatment of prostate mucosa adenocarcinoma under multidisciplinary diagnosis and treatment mode: 2 cases report and literature review
Peng WU ; Fuli WANG ; Jing ZHANG ; Jing REN ; Zhiyong QUAN ; Wanni XU ; Lichun WEI ; Weijun QIN
Journal of Modern Urology 2024;29(2):154-157
【Objective】 To explore the clinicopathological characteristics and comprehensive treatment strategies of prostate mucosa adenocarcinoma under multidisciplinary diagnosis and treatment (MDT) mode. 【Methods】 Data of two patients with typical prostate mucosa adenocarcinoma treated in our hospital during Sep.2020 and Apr.2023 were retrospectively analyzed. 【Results】 In case 1, the clinical manifestation was macroscopic hematuria; multiparametric magnetic resonance imaging (mpMRI) indicated solid prostatic nodules, clinical stage T4N1Mx; initial prostate specific antigen (PSA) was 1.2 ng/mL; 6868Ga-prostate specific membrane antigen PET/CT (68Ga-PSMA PET/CT) suggested abnormal uptake of nuclear lesions in the prostate (SUV4.2-5.3); biopsy results indicated invasive mucinous adenocarcinoma.After prostate and pelvic field radiotherapy + androgen deprivation therapy (ADT) + antihypertensive treatment, lesions were significantly reduced, and hematuria symptoms were relieved.In case 2, the clinical manifestation was dysuria; initial PSA was 91.78 ng/mL; mpMRI suggested invasion of prostate mass into the bladder and clinical stage of T4N1M1b; 68Ga-PSMA PET/CT indicated prostate and pelvic lymph nodes, and multiple bone lesions showed increased nuclide uptake; biopsy results indicated prostate adenocarcinoma with mucinous adenocarcinoma.Initial endocrine treatment was performed.After 3 months, PSA was reduced to 0.083 ng/mL, and imaging showed the tumor was significantly reduced.Robotic-assisted laparoscopic tumor prostatectomy with extended pelvic lymph node dissection was performed, and endocrine adjuvant therapy was continued after surgery. 【Conclusion】 Prostate mucosa adenocarcinoma has different clinicopathological characteristics and prognosis from conventional acinar adenocarcinoma, and the whole-process management under MDT mode is of great value in the diagnosis and treatment of this disease.
10.Effects of PATL1 on the proliferation and migration of gastric cancer cells and its mechanism of action
Zeyu Xu ; Zhiyong Lai ; Yifan Ren ; Feng Wu ; Yuting Peng ; Jun Xu
Acta Universitatis Medicinalis Anhui 2024;59(1):133-139
Objective :
Objective
Methods :
The expression levels of PATL1 in pancar- cinoma,gastric cancer and normal tissues were analyzed by TCGA database.The expression level of PATL1 in 40 human gastric cancer tissues and paired adjacent tissues was evaluated by quantitative real-time PCR (RT-qPCR) . The Kaplan-Meier Plotter database was used to analyze the prognosis of PATL1 in gastric cancer patients.The gas- tric cancer cell line AGS was transfected with PATL1 interference vector,and the interference effect was evaluated by RT-qPCR. The effects of PATL1 on the proliferation and migration of AGS were detected by cell counting kit-8 ( CCK-8) ,Transwell test and scratch healing test.The effects of interference with PATL1 on the expression of cel- lular-myelocytomatosis viral oncogene ( c-Myc) and autophagy related 7 ( ATG7) proteins in gastric cancer cells were detected by Western blot assay.
Results :
RT-qPCR showed that the expression of PATL1 in human gastric cancer tissue was higher than that in normal gastric tissue (P<0. 001) ,and PATL1 was correlated with the progno- sis of patients with enteric gastric cancer (P<0. 000 1) .After PATL1 was knocked down,the number of prolifera- ting and migrating gastric cancer cells decreased (P<0. 05) .Western blot test results showed that the expression level of ATG7 protein decreased after PATL1 was knocked down (P<0. 05) .
Conclusion
PATL1 may inhibit the proliferation and migration of gastric cancer cells through crosstalk with c-Myc and ATG7 .


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