1.Hapln1 promotes dedifferentiation and proliferation of iPSC-derived cardiomyocytes by promoting versican-based GDF11 trapping
Hao DING-JUN ; Qin YUE ; Zhou SHI-JIE ; Dong BU-HUAI ; Yang JUN-SONG ; Zou PENG ; Wang LI-PING ; Zhao YUAN-TING
Journal of Pharmaceutical Analysis 2024;14(3):335-347
Hyaluronan and proteoglycan link protein 1(Hapln1)supports active cardiomyogenesis in zebrafish hearts,but its regulation in mammal cardiomyocytes is unclear.This study aimed to explore the potential regulation of Hapln1 in the dedifferentiation and proliferation of cardiomyocytes and its therapeutic value in myocardial infarction with human induced pluripotent stem cell(hiPSC)-derived car-diomyocytes(CMs)and an adult mouse model of myocardial infarction.HiPSC-CMs and adult mice with myocardial infarction were used as in vitro and in vivo models,respectively.Previous single-cell RNA sequencing data were retrieved for bioinformatic exploration.The results showed that recombinant human Hapln1(rhHapln1)promotes the proliferation of hiPSC-CMs in a dose-dependent manner.As a physical binding protein of Hapln1,versican interacted with Nodal growth differentiation factor(NODAL)and growth differentiation factor 11(GDF11).GDF11,but not NODAL,was expressed by hiPSC-CMs.GDF11 expression was unaffected by rhHapln1 treatment.However,this molecule was required for rhHapln1-mediated activation of the transforming growth factor(TGF)-β/Drosophila mothers against decapentaplegic protein(SMAD)2/3 signaling in hiPSC-CMs,which stimulates cell dedifferentiation and proliferation.Recombinant mouse Hapln1(rmHapln1)could induce cardiac regeneration in the adult mouse model of myocardial infarction.In addition,rmHapln1 induced hiPSC-CM proliferation.In conclusion,Hapln1 can stimulate the dedifferentiation and proliferation of iPSC-derived cardiomyocytes by promoting versican-based GDF11 trapping and subsequent activation of the TGF-β/SMAD2/3 signaling pathway.Hapln1 might be an effective hiPSC-CM dedifferentiation and proliferation agent and a po-tential reagent for repairing damaged hearts.
2.Mechanism of Kechuanting granules in suppressing IL-33/ILC2s and pathogenic T cells to intervene in allergic airway inflammation
Nan-Ting ZOU ; Zhao WU ; Xiao-Dong YAN ; Chun-Fei ZHANG ; Hao-Hong ZHANG ; Qing-Yan MO ; Ming-Qian JU ; Jin-Zhu XU ; Chun-Ping WAN
Chinese Pharmacological Bulletin 2024;40(7):1350-1357
Aim To investigate the mechanisms of Ke-chuanting granules(KCT)inhibiting the IL-33/ILC2s pathway and pathogenic T cells to intervene in allergic airway inflammation.Methods Network pharmacolo-gy was utilized to analyze the potential targets and mechanisms of KCT-treated asthma.Allergic asthma models were induced in mice using OVA.Lung histo-pathology was conducted to observe injury changes.ELISA and quantitative PCR were utilized to measure key inflammatory factors and their mRNA expression levels in Th2-type asthma.Western blot was used to detect the phosphorylation levels of relevant proteins in the MAPK pathway.Flow cytometry was performed to evaluate the proportions of ILC2s,Th1,Th 17,Th2 and Treg cells.Results Network pharmacology iden-tified 227 main active components and 143 key targets of KCT in treating asthma,primarily enriched in signa-ling pathways such as MAPK and IL-17.Further vali-dation experiments demonstrated that KCT significantly alleviated lung inflammatory injury in asthmatic mice,reduced the number of B cells,production of I L-4,TNF-α and TGF-β,downregulated JNK phosphoryla-tion levels in lung tissue,as well as mRNA levels of Il-33,Bcl11b,Rorα,Tcf-7,Jun,Mapk3 and Mapk14.KCT intervention reduced the numbers of ILC2s and Th 17 cells in lungs and spleens of mice,and inhibited Th2 cell infiltration in lungs.Conclusions KCT ex-hibits therapeutic effects on allergic airway inflamma-tion in asthma,closely associated with the inhibition of the IL-33/ILC2s pathway,pathogenic T cell subsets,and JNK-MAPK signaling pathway.
3.Regulatory effect of five-element music therapy of traditional Chinese medicine for suboptimal health status of liver stagnation and spleen deficiency based on the temperature changes of acupoints.
Jing-Yu ZHANG ; Bo JI ; Yi-Tian LIU ; Yang FANG ; Tian-Yu SHI ; Qian-Wei LIU ; Yun-Peng GE ; Jia-Jia WANG ; Zi-Yue ZHANG ; Na-Na HAN ; Yu-Ting LIU ; Shi-Dong ZOU
Chinese Acupuncture & Moxibustion 2022;42(11):1253-1259
OBJECTIVE:
To observe the effect of five-element music therapy of traditional Chinese medicine (TCM) on the clinical symptoms and the quality of life in the patients with suboptimal health status (SHS) of liver stagnation and spleen deficiency and explore the corresponding specificity changes in the temperature of acupoints when zangfu functions are of dysfunction and recovered to be balanced, separately.
METHODS:
Sixty patients with SHS of liver stagnation and spleen deficiency were randomized into an observation group and a control group, 30 cases in each one. In the control group, the conventional health education was provided. In the observation group, on the base of the therapeutic regimen as the control group, the patients received the five-element music therapy to pacify the liver qi and strengthen the spleen functions, once every two days, 30 min each time, 3 treatments a week. The course of treatment consisted of 4 weeks. Before and after treatment, the TCM syndrome score and the MOS 36-item short form healthy survey (SF-36) score were compared between the two groups and the clinical therapeutic effect was evaluated. Using infrared thermal imaging, the temperature at the acupoints of the affected organs (liver, spleen), the related organs (gallbladder, stomach) and the other non-related zangfu organs (pericardium, lung) was detected before and after treatment in the two groups.
RESULTS:
After treatment, the TCM syndrome scores were reduced when compared with those before treatment in the two groups (P<0.01, P<0.05); the reduction in the observation group was larger than that of the control group (P<0.01). The score of each domain for the SF-36 in the observation group and the score of role-emotional domain in the control group were all increased when compared with the scores before treatment (P<0.01, P<0.05); and the score of each domain for the SF-36 in the observation group was higher than that in the control group (P<0.01). The total effective rate was 66.7% (20/30) in the observation group, which was higher than 10.0% in the control group (3/30, P<0.05). In the observation group, the temperature of the yuan-primary point, the back-shu point and the front-mu point related to the liver, as well as those related to the gallbladder after treatment was reduced when compared with the temperature before treatment; and the changes were larger than those of the control group (P<0.01). The temperature of the yuan-primary point, the back-shu point and the front-mu point related to the spleen, as well as the back-shu point and the front-mu point related to the stomach in the observation group was increased when compared with the temperature before treatment (P<0.01); and the changes were larger than those of the control group (P<0.01). For the temperature of the non-specific points related to the liver and spleen, as well as the yuan-primary point, the back-shu point and the front-mu point related to the pericardium and the lung, there was no significant differences when compared with the temperature at the above-mentioned acupoints before and after treatment (P>0.05).
CONCLUSION
TCM five-element music therapy associated with the conventional health education may effectively relieve the clinical symptoms and improve the quality of life in the patients with suboptimal health status of liver stagnation and sleep deficiency; and the therapeutic effect is better than the simple health education. The changes in the temperature of acupoints may reflect the functional regulation of the related zangfu organs in the body.
Humans
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Acupuncture Points
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Medicine, Chinese Traditional
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Spleen
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Music Therapy
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Quality of Life
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Temperature
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Liver
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Health Status
4. Determination of prucalopride in human plasma by LC-MS/MS analysis
Chang-Yin LI ; Jian-Cheng LIAO ; Ming-Xia LU ; Hui-Ting SONG ; Wen-Zheng JU ; Jian-Dong ZOU ; Ji-Hong CHU
Chinese Pharmacological Bulletin 2022;38(5):789-794
Aim To develop an LC-MS/MS method for the determination of prucalopride(PCP)in human plasma.Methods Prucalopride -13CD3(dPCP)was used as the internal standard.The analytes were extracted from human plasma through liquid-liquid extraction method using ethyl acetate, followed by being dried, and then the reconstitution was injected into LC-MS/MS systems.Agilent ZORBAX SB C18(3.0×100 mm, 3.5 μm)column and isocratic elution system composing of methanol and 1 mmol·L-1 ammonium acetate(80:20, V/V)provided chromatographic separation of PCP and dPCP.AB Sciex API4000 mass spectrometer equipped with an electrospray ionization source in positive ion mode was employed for mass detection, and data acquisition was carried out in multiple reaction monitoring(MRM)mode.The mass transition ion-pair was followed as m/z 368.4/196.0 for PCP and m/z 374.4/198.0 for dPCP.Results PCP and dPCP were eluted at 3.6 min, with no interference in human blank plasma.PCP in human plasma showed good linearity over the concentration range of 0.058 96-7.547 μg·L-1 with the correlation coefficient of 0.996 3-0.999 6.The lower limit of quantitation of this method was 0.058 96 μg·L-1.The intra-batch and inter-batch accuracy ranged from 98.29% to 108.2%, with good precision(CV<5.2%).The average matrix factors of normal, haemolysed and lipaemic matrix human samples all ranged from 96.48% to 106.3% with CV less than 8.39%.The average extraction recoveries of PCP at low, medium and high concentrations were 89.88%, 95.27% and 94.52% respectively, with CV less than 7.21%.PCP was stable in human samples after 6 h at room temperature, 60 h at -20 ℃, 56 days or three freeze-thaw cycles at -80 ℃; meanwhile, the processed plasma samples remained stable after being stored for 24 hours in autosampler at 8 ℃.Furthermore, PCP in human blood samples was proved to be stable after 4 h at room temperature.Conclusions The present LC-MS/MS method for the determination of PCP in human plasma was convenient, accurate, sensitive, stable, specific and reproducible and was proved to be suitable for the clinical pharmacokinetics and bioequivalence studies of PCP preparations.
5.Nutritional status and influencing factors in the elderly at home
ZHAO Dong ; SU Dan Ting ; HUANG Li Chun ; CHEN Qi ; ZHU Zhi Hong ; LIU Sheng ; GU Wei ; ZOU Yan
Journal of Preventive Medicine 2021;33(5):468-472
Objective:
To learn the results of MNA ( mini nutritional assessment ) nutrition screening and influencing factors in the elderly living at home, so as to provide basis for improving the nutritional status of the elderly living at home.
Methods:
The elderly people at home were recruited from Yinzhou District, Yiwu City and Changshan County in Zhejiang Province by the multi-stage random sampling method. Their demographic information, living habits and nutritional status were collected by the MNA scale and the questionnaire for nutrition and health status surveillance. The multivariate linear regression model was used to analyze influencing factors for the nutritional status.
Results:
Of 374 study subjects, 186 ( 49.73% ) were males and 188 ( 50.27% ) were females. The age was ( 69.63±6.68 ) years ( range, 60-90 years ). The average score of MNA scale was 25.26±2.81. The prevalence of malnutrition risk in the elderly living at home was 20.59%. Age ( β'=-0.140), marital status ( β'=0.110 ), annual income ( β'=0.155 ), active physical exercise ( β'= 0.104 ), eating health products/nutritional supplements ( β'= 0.110 ) and satiety ( full diet β'=0.196 ) were influencing factors for MNA scores ( P<0.05 ).
Conclusion
The prevalence of malnutrition risk among the elderly living at home is 20.59%. The prevalence increases with age. Having a spouse, doing active physical exercise, eating health products/nutritional supplements, having healthy eating habits are conducive to maintaining the nutritional health of the elderly.
6.Professor
Dong-Dong WANG ; Bo LI ; Yong-Mei ZHA ; Han ZOU ; Ting-Ting YAO ; Wen GU ; Jun YANG ; Qing-Ping ZHANG
Chinese Acupuncture & Moxibustion 2021;41(3):313-315
Professor
Acupuncture
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Acupuncture Points
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Acupuncture Therapy
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Facial Paralysis/therapy*
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Humans
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Meridians
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Moxibustion
7.Discrimination of Armeniacae Semen Amarum from different processed products and various rancidness degrees by electronic nose and support vector machine.
Jian-Ting GONG ; Li-Ying ZHAO ; Dong XU ; Jia-Hui LI ; Xin CHEN ; Hui-Qin ZOU ; Yong-Hong YAN
China Journal of Chinese Materia Medica 2020;45(10):2389-2394
This study was aimed to develop a simple, rapid and reliable method for identifying Armeniacae Semen Amarum from different processed products and various rancidness degrees. The objective odor information of Armeniacae Semen Amarum was obtained by electronic nose. 105 batches of Armeniacae Semen Amarum samples were studied, including three processed products of Armeniacae Semen Amarum, fried Armeniacae Semen Amarum and peeled Armeniacae Semen Amarum, as well as the samples with various rancidness degrees: without rancidness, slight rancidness, and rancidness. The discriminant models of different processed products and rancidness degrees of Armeniacae Semen Amarum were established by Support Vector Machine(SVM), respectively, and the models were verified based on back estimation of blind samples. The results showed that there were differences in the characteristic response radar patterns of the sensor array of different processed products and the samples with different rancidness degrees. The initial identification rate was 95.90% and 92.45%, whilst validation recognition rate was 95.38% and 91.08% in SVM identification models. In conclusion, differentiation in odor of different processed and rancidness degree Armeniacae Semen Amarum was performed by the electronic nose technology, and different processed and rancidness degrees Armeniacae Semen Amarum were successfully discriminated by combining with SVM. This research provides ideas and methods for objective identification of odor of traditional Chinese medicine, conducive to the inheritance and development of traditional experience in odor identification.
Drugs, Chinese Herbal
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Electronic Nose
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Medicine, Chinese Traditional
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Semen
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Support Vector Machine
8.Change pattern and correlation analysis of macroscopic characteristics, active components and rancidness degrees of Armeniacae Semen Amarum in deterioration process.
Jian-Ting GONG ; Li-Ying ZHAO ; Dong XU ; Yue-Bao YAO ; Bin-Qing LIU ; Hui-Qin ZOU ; Yong-Hong YAN
China Journal of Chinese Materia Medica 2020;45(13):3155-3160
To discuss the effect of deterioration on the quality of Armeniacae Semen Amarum by observing the changes of macroscopic characteristics, active components and rancidness degrees of Armeniacae Semen Amarum in deterioration process. The traditional macroscopic identification was used to observe, identify and classify the morphologic and organleptic characteristics of Armeniacae Semen Amarum. The contents of amygdalin and fatty oil(two representatives of active components) were detected by HPLC and general rule 0713 in Chinese Pharmacopoeia, respectively. Acid value and peroxide value of the samples were selected as the representative indices of different rancidness degrees, and the general rule 2303 was adopted as the method for quantitative analysis. Then principal component analysis(PCA), partial least square analysis discrimination analysis(PLS-DA) were further utilized to establish the discriminative models of samples with different rancidness degrees, and also to screen out the largest contribution factors. In sensory evaluation, Armeniacae Semen Amarum samples were divided into three groups: non-rancid, slightly-rancid, and noticeably-rancid. The color of seed coat, cotyledon and surface of noticeably-rancid samples was deepened, and the odor differed much from non-rancid samples. Average content of amygdalin and fatty oil in non-rancid samples was 4.12% and 67.77%, respectively, both meeting the requirements of Chinese Pharmacopoeia; and decreased to some extent in slightly-rancid samples. However, the content of amygdalin sharply dropped to 0.074% in noticeably-rancid samples. The acid value and peroxide value were increased significantly with the intensifying of the rancidness degree, from only 1.363 and 0.016 74 in non-rancid samples to 1.865 and 0.023 70 in slightly-rancid samples, even doubled in noticeably-rancid samples(2.167 and 0.033 82). The discriminative models established by PCA and PLS-DA could complete the task of distinguishing the non-rancid samples from noticeably-rancid ones. The contribution degree of amygdalin content as one of the input attributes of discriminative model was higher than 1. Rancidness affected the quality of Armeniacae Semen Amarum, resulting in appearance changes, decrease in content of active components, and increase in acid value and peroxide value. Obviously, noticeably-rancid samples were non-conforming to Chinese Pharmacopoeia and no longer suitable for medicinal use. Rancidness can significantly reduce the quality of Armeniacae Semen Amarum, and even could possibly produce toxicity, which should attach more attention.
Amygdalin
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Chromatography, High Pressure Liquid
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Drugs, Chinese Herbal
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Semen
9.Analysis of Driver-Passenger Relationship and Restoration of Accident Process Based on 3D Laser Scanning Technology.
Xue ZHANG ; Sheng Dong LI ; Yan Geng YU ; Fu ZHANG ; Chong Bin ZOU ; Zhi Liang ZOU ; Qi Feng MIAO ; Man Ting ZHANG ; Li Juan TANG ; Dong Ri LI
Journal of Forensic Medicine 2020;36(1):69-71
Objective To discuss the application of 3D laser scanner and computer technology in restoration of the accident scene and reconstruction of the accident process, as well as identification of the driver-passenger relationship. Methods The scene of a traffic accident, the accident vehicle and the vehicle of the same type as accident vehicle were scanned using 3D laser scanner. The accident scene, traces and accident vehicle were integrated using computer technology to restore the accident scene, and the accident process was reconstructed and analyzed by combining the characteristics of the body injuries. Results By restoring the accident scene and reconstructing the accident process with 3D laser scanner, it was determined that Wu was in the driving seat at the time of the accident. Conclusion It is more objective and scientific to use 3D laser scanning technology to restore the accident scene, reconstruct the accident process and analyze the moving track of the driver and passengers in the vehicle. It will help to improve the accuracy of forensic identification of road traffic accidents.
Accidents, Traffic
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Computer Simulation
10.INI1 Induces Adipocytic Differentiation of Epithelioid Sarcoma Cells
Xian-biao XIE ; Li-li WEN ; Dong-ming LV ; Yu-tong ZOU ; Hao YAO ; Ting-sheng PENG
Journal of Sun Yat-sen University(Medical Sciences) 2020;41(5):690-696
【Objective】 To study the function and mechanism of INI1 in human epithelioid sarcoma cells. 【Methods】 Western blotting was used to test the expressionlevel of INI1in epithelioid sarcoma cells. The expression of INI1 in clinical specimens of epithelioid sarcoma was detected by immunohistochemistry. Tet-on system was used to establish the expression of INI1 in epithelioid sarcoma cells. Following the induction of INI1 expression, the cell morphology, proliferation and the molecular markers of adipocytic differentiation were detected. Then western blot was employed to detect the transcription factors of adipocytic differentiation, and oil red O staining was also tested. 【Results】 The expression of INI1 was absent in both epithelioid sarcoma cells and clinical tissues(P<0.05). The INI1 expression in the epithelioid sarcoma cells was successfully established by Tet-on system. After induction by doxycycline, the expression of INI1 was up-regulated. With continuous expression of INI1, the morphology of VA-ES-BJ cells was changed, with the appearance of abundant vacuoles in the cytoplasm. The cell proliferation was obviously inhibited(P<0.05). Epithelial cell marker Cytokeratin was found significantly reduced(P<0.05). In addition, the adipocyte markers, leptin, adiponectin and lipoprotein lipase were significantly up-regulated(P<0.05). The adipogenic transcriptional factors PPAR-γ and CEBP-α were found upregulated(P<0.05) and the oil red staining was significantly positive. 【Conclusions】 The expression of INI1 was absent in epithelioid sarcoma. Reconstruction of the expression of INI1 could induce adipocytic differentiation via upregulation of transcriptional factors PPAR-γand CEBP-αin epithelioid sarcoma cells.


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