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.Reference values for inspiratory muscle related indexes of preschool children in Nanjing
Yanlu HE ; Zhiyong YANG ; Weiding CHEN ; Xia CHI ; Meiling TONG
Chinese Journal of Child Health Care 2024;32(5):507-510
【Objective】 To generate reference values for inspiratory muscle of preschool children in Nanjing, so as to provide a reference index for evaluating children′s lung function and exercise performance. 【Methods】 A total of 236 preschool children were selected from the main urban area of Nanjing by stratified cluster sampling.The inspiratory muscles were evaluated by breath link respiratory function evaluation system to obtain the maximum inspiratory pressure (MIP) and inspiratory peak flow rate.Pearson analysis was used to determine the correlation of MIP and inspiratory peak flow rate with gender, age, height and weight.Multiple linear stepwise regression analysis was used to obtain the formula of MIP and inspiratory peak flow rate. 【Results】 Differences in MIP and peak inspiratory flow rate were not significant between boys and girls (P>0.05), but were statistically significant among different age groups and showed an increasing trend with age (F=13.660, 33.581, P<0.001).MIP and peak inspiratory flow rate were positively correlated with children′s age, height and weight (P<0.001).The regression model, proved to be statistically significant(F=12.913、22.398, P<0.08), indicated that height was the best predictor of MIP and age was the best predictor of inspiratory peak flow rate. 【Conclusions】 This study is the first study on the predicted value of inspiratory muscle in preschool children in China.The predicted value formula can provide a reference for clinical inspiratory muscle evaluation.
3.Experts consensus on standard items of the cohort construction and quality control of temporomandibular joint diseases (2024)
Min HU ; Chi YANG ; Huawei LIU ; Haixia LU ; Chen YAO ; Qiufei XIE ; Yongjin CHEN ; Kaiyuan FU ; Bing FANG ; Songsong ZHU ; Qing ZHOU ; Zhiye CHEN ; Yaomin ZHU ; Qingbin ZHANG ; Ying YAN ; Xing LONG ; Zhiyong LI ; Yehua GAN ; Shibin YU ; Yuxing BAI ; Yi ZHANG ; Yanyi WANG ; Jie LEI ; Yong CHENG ; Changkui LIU ; Ye CAO ; Dongmei HE ; Ning WEN ; Shanyong ZHANG ; Minjie CHEN ; Guoliang JIAO ; Xinhua LIU ; Hua JIANG ; Yang HE ; Pei SHEN ; Haitao HUANG ; Yongfeng LI ; Jisi ZHENG ; Jing GUO ; Lisheng ZHAO ; Laiqing XU
Chinese Journal of Stomatology 2024;59(10):977-987
Temporomandibular joint (TMJ) diseases are common clinical conditions. The number of patients with TMJ diseases is large, and the etiology, epidemiology, disease spectrum, and treatment of the disease remain controversial and unknown. To understand and master the current situation of the occurrence, development and prevention of TMJ diseases, as well as to identify the patterns in etiology, incidence, drug sensitivity, and prognosis is crucial for alleviating patients′suffering.This will facilitate in-depth medical research, effective disease prevention measures, and the formulation of corresponding health policies. Cohort construction and research has an irreplaceable role in precise disease prevention and significant improvement in diagnosis and treatment levels. Large-scale cohort studies are needed to explore the relationship between potential risk factors and outcomes of TMJ diseases, and to observe disease prognoses through long-term follw-ups. The consensus aims to establish a standard conceptual frame work for a cohort study on patients with TMJ disease while providing ideas for cohort data standards to this condition. TMJ disease cohort data consists of both common data standards applicable to all specific disease cohorts as well as disease-specific data standards. Common data were available for each specific disease cohort. By integrating different cohort research resources, standard problems or study variables can be unified. Long-term follow-up can be performed using consistent definitions and criteria across different projects for better core data collection. It is hoped that this consensus will be facilitate the development cohort studies of TMJ diseases.
4.TCOF1 Gene variation in Treacher Collins syndrome and evaluation of speech rehabilitation after bone bridge surgery.
Yonghua LI ; Wenyue CHI ; Ken LIN ; Jinyan ZU ; Hua SHAO ; Zhiyong MAO ; Quandong CHEN ; Jing MA
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2023;37(9):748-754
Objective:By analyzing the clinical phenotypic characteristics and gene sequences of two patients with Treacher Collins syndrome(TCS), the biological causes of the disease were determined. Then discuss the therapeutic effect of hearing intervention after bone bridge implantation. Methods:All clinical data of the two family members were collected, and the patients signed the informed consent. The peripheral blood of the proband and family members was extracted, DNA was extracted for whole exome sequencing, and Sanger sequencing was performed on the family members for the mutation site.TCOF1genetic mutations analysis was performed on the paitents. Then, the hearing threshold and speech recognition rate of family 2 proband were evaluated and compared under the sound field between bare ear and wearing bone bridge. Results:In the two pedigrees, the probands of both families presented with auricle deformity, zygomatic and mandibular hypoplasia, micrognathia, hypotropia of the eye fissure, and hypoplasia of the medial eyelashes. The proband of Family 1 also presents with specific features including right-sided narrow anterior nasal aperture and dental hypoplasia, which were consistent with the clinical diagnosis of Treacher Collins syndrome. Genetic testing was conducted on both families, and two heterozygous mutations were identified in the TCOF1 gene: c. 1350_1351dupGG(p. A451Gfs*43) and c. 4362_4366del(p. K1457Efs*12), resulting in frameshift mutations in the amino acid sequence. Sanger sequencing validation of the TCOF1 gene in the parents of the proband in Family 1 did not detect any mutations. Proband 1 TCOF1 c. 1350_1351dupGG heterozygous variants have not been reported previously. The postoperative monosyllabic speech recognition rate of family 2 proband was 76%, the Categories of Auditory Performance(CAP) score was 6, and the Speech Intelligibility Rating(SIR) score was 4. Assessment using the Meaningful Auditory Integration Scale(MAIS) showed notable improvement in the patient's auditory perception, comprehension, and usage of hearing aids. Evaluation using the Glasgow Children's Benefit Inventory and quality of life assessment revealed significant improvements in the child's self care abilities, daily living and learning, social interactions, and psychological well being, as perceived by the parents. Conclusion:This study has elucidated the biological cause of Treacher Collins syndrome, enriched the spectrum of TCOF1 gene mutations in the Chinese population, and demonstrated that bone bridge implantation can improve the auditory and speech recognition rates in TCS patients.
Child
;
Humans
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Mandibulofacial Dysostosis/genetics*
;
Quality of Life
;
Speech
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Parents
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Mutation
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Nuclear Proteins/genetics*
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Phosphoproteins/genetics*
5.Analysis of diagnosis and treatment of adrenal lymphangioma
Dianbin SONG ; Jingjing ZHANG ; Man YU ; Qiang CHI ; Hui XU ; Yan MAO ; Zhiyong WANG
Chinese Journal of Urology 2021;42(2):144-146
By summarizing and analyzing the clinical diagnosis and treatment experience of 17 cases of adrenal lymphangioma, the imaging characteristics and pathological types of the disease were discussed. The results showed that the imaging of adrenal lymphangioma was non-specific, and the appearance was similar to that of general cysts.Howerer, the density of the cyst was slightly higher than that of simple cysts. Some cases showed calcification on the cyst wall, and a few showed adenoma-like appearance. The diagnosis mainly depends on pathological examination. For those patients with tumors ≥4.0 cm, endocrine function, suspected malignancy, or obvious clinical symptoms, surgery is recommended.
6.One case report of adrenal enteric cyst
Dianbin SONG ; Jingjing ZHANG ; Qiang CHI ; Hui XU ; Hongyang LI ; Zhiyong WANG
Chinese Journal of Urology 2020;41(9):705-706
A patient with adrenal enteric cyst was reported. The patient was admitted to the hospital for physical examination and found adrenal glands. The preoperative diagnosis considered simple adrenal cysts and underwent complete surgical resection. Postoperative pathological returns were adrenal enteric cysts. There was no recurrence at 1 year postoperative follow-up and recovery was good. The clinical symptoms of the disease are not typical. The imaging and laboratory indicators are not specific. Pathological diagnosis is required, and surgical resection is the best treatment plan.
7.Comparison of social function, adverse reaction and medication adherence of Paliperidone, Amisulpride and Olanzapine in patients with first-episode schizophrenia
Zhiyong LAN ; Weiming HU ; Chi ZHANG ; Lifeng ZHENG ; Xiaofeng GAO ; Wuyuan LAN
Journal of Chinese Physician 2018;20(2):216-219
Objective To investigate the social function, adverse reaction and medication adherence of paliperidone, amisulpride, and olanzapine in patients with first episode schizophrenia.Methods A total of 96 patients with first episode of schizophrenia was randomly divided into three groups, with reference to random numbers, among which there were 32 in paliperidone group, 32 cases in amisulpride group, and 32 in olanzapine group.All the patients in all groups were assessed with negative and positive scale (PANSS), personal and social performance scale (PSP), drug attitude inventory (DAI) at baseline and the end of 6 months.Results (1) There was no significant difference in the therapeutic effect between three groups (P > 0.05);(2) The scores of PSP and DAI were increased in three groups after treatment, and the difference was statistically significant compared to that before treatment (P < 0.05).Conclusions Three drugs have similar efficacy in the treatment of first-episode schizophrenia, and there is no significant difference in improving medication compliance and social function.
8.Effects of TLR4?mediated Cx43 expression on proliferation and apoptosis of bladder cancer cells by LPS
Qiang CHI ; Xiaolin AI ; Zhiyong WANG ; Guang MA ; Ying LIU ; Hui XU
The Journal of Practical Medicine 2017;33(5):701-705
Objective To study the impacts of lipopolysaccharide on expressions of Cx43 and TLR4 proteins in bladder cancer cell lines and on cell proliferation and apoptosis. Methods 5637 cells were cultured in vitro. After stimulation with LPS,expressions of Cx43 and TLR4 proteins were detected by Western blot in bladder cancer 5637 cell. The proliferation and apoptosis in the 5637?control group,5637?LPS group,and 5637?LPS +cisplatin group were detected by CCK?8(cell counting kit)and flow cytometry. Results The gene expression of Cx43 in the 5637?LPS group was significantly higher than that in the 5637?control group(t=3.892,P=0.012). The expressions of TLR4 and Cx43 in 5637?LPS group were significantly higher than those in the 5637?control group(t=7.029,P=0.019;and t=18.17,P=0.003). The proliferation was significantly decreased in the 5637?control group as compared with the 5637?LPS group (t = 8.756,P = 0.018). The apoptotic rate was (8.3 ± 1.58)% in the 5637?control group and (7.8 ± 2.03)% in the 5637?LPS group,with no significant statistical difference(t = 2.935,P = 0.099). However,the rate of the 5637?cisplatin group(60 ± 4.35)%was higher than that in 5637?cisplatin + LPS group(52 ± 6.25)%,the difference was statistically significant(t = 6.992,P =0.019). Conclusions Under the stimulation of LPS,bladder tumor cells may induce tumor cells to escape immune surveillance by increasing the expressions of TLR4 and Cx43.
9.Application value of urine kidney injury molecule-1 in early evaluating the renal function for unilateral obstruction renal disease
Zhiyong WANG ; Li GUO ; Hongyang LI ; Xiuming LI ; Qiang CHI
Chinese Journal of Postgraduates of Medicine 2017;40(11):978-981
Objective To study the early renal function assessment value of urine kidney injury molecule-1(KIM-1)level in patients with unilateral obstruction renal diseases.Methods Seventy-eight patients with unilateral obstruction renal diseases were selected,and the glomerular filtration rate(GFR) was detected by 99Tcm-diethylene triaminepentaacetic acid (DTPA) renal dynamic imaging method. The patients were divided into 4 groups according to the GFR: normal renal function group with GFR>40 ml/(min·1.73 m2)25 cases(A group),low-injury renal function group with 20 ml/(min·1.73 m2)
10.Expression and significance of the proteins in TSP-1 and NF-κB signal pathways of infantile capillary hemangioma
Yongting ZHANG ; Weili XU ; Suolin LI ; Zhiyong ZHONG ; Yingchao LI ; Wenbo WANG ; Chi SUN
Chinese Journal of Plastic Surgery 2016;32(6):441-446
Objective To explore the significance of the relative proteins in thrombin sensitive protein(TSP-1) and nuclear factor kappa B (NF-κB) signal pathways by detecting their expressions in different phases of hemangioma and the relations between them and microvessel density(MVD).Methods Forty cases with hemangioma were randomly selected and divided into proliferative group(n =21) and involutional group(n =19) according to Mulliken standard.The expressions of TSP-1,CD36,p59fyn,Caspase-3,p38MAPK,NF-κBp65,p-IκBα,p-IKKβ,VEGF and CD34 in endothelial cells were detected by immunohistochemistry.The relevance between the proteins in TSP-1 and NF-κB signal pathways and MVD were compared and analyzed respectively.Results The expressions of TSP-1,CD36,p59fyn,Caspase-3,p38MAPK in proliferative hemangioma were all lower than those in involutional hemangioma (F=32.582,47.575,11.645,18.824,13.140;P =0.000,0.000,0.002,0.000,0.001).Conversely,the expressions of NF-κBp65,p-IκBα,p-IKKβ,VEGF in proliferative hemangioma were higher than those in involutional hemangioma(F =7.807,6.323,5.988,4.216;P =0.009,0.018,0.021,0.049).Meanwhile,MVD were different between the two groups(F =7.256,P =0.012).The expressions of TSP-1,CD36,p59fyn and MVD showed a negative relationship(rs =-0.420,-0.519,-0.388;P =0.021,0.003,0.034).Conversely,the expressions of NF-κBp65,pvIκBα,VEGF and MVD showed a positive relationship(rs =0.666,0.611,0.515;P =0.000,0.000,0.004).The expression of TSP-1 and NF-κBp65 showed a negative relationship (rs =-0.413,P =0.023).Conclusions TSP-1 signal pathway and NF-κBp65 signal pathway may be interactive in evolution of infantile hemangioma,which has a potential role by effecting the progress of angiogenesis.

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