1.PIEZO1 Channel is Involved in Electric Field Guided Cell Migration
Gui-Cheng ZHANG ; Peng GONG ; Yi-Fan WANG ; San-Jun ZHAO
Progress in Biochemistry and Biophysics 2024;51(3):673-684
ObjectiveDisruption of epithelial layer may instantaneously induce the generation of endogenous electric fields, which was proved to play an important role in guiding the cell migration and promoting wound healing. PIEZO1 is a kind of mechanic sensitive channel, may be regulated by voltage, is proved to involve in chemotactic migration of cells and play an important role in the process of wound healing. In this paper, the role of PIEZO1 and its downstream proteins FAK and integrin β1 in the electric field guided cell migration were investigated by HaCaT cells (human immortalized keratinocyte). MethodsCell migration was tracked by Living Cell Imaging System in directed current (DC) electric field (EF). Inhibitors and RNAi techniques were applied to study the function of PIEZO1 and other related proteins in electric fields. Western blot was used to detect the expression and phosphorylation levels of integrin β1 and FAK in electric field guided migration under EF stimulation. ResultsPiezo1 RNAi as well as Ruthenium red and GsMTx4 treatment all significantly inhibited the electrotaxis of HaCaT cells. Electric field stimulation with GsMTx4 treatment alone increased FAK phosphorylation level and the expression of integrin β1. Electric field promoted the expression level of integrin β1 and the phosphorylation level of FAK. Inhibiting the expression of PIEZO1 by RNAi significantly attenuated the phosphorylation level of FAK under EF stimulation. Inhibition of integrin β1 and FAK by inhibitor significantly decrease the electric field guided cell migration. ConclusionPIEZO1 as well as integrin β1 and FAK are involved in the electric field guided cell migration of HaCaT cells. Electric field signals regulate the expression of integrin β1 and the activation of FAK through PIEZO1-mediated signal pathway to orchestrate cell migration.
2.Molecular cloning and characterization of three phenylalanine ammonia-lyase genes from Schisandra chinensis.
San-Peng FAN ; Wei CHEN ; Jiang-Chun WEI ; Xiao-Xu GAO ; Yong-Cheng YANG ; An-Hua WANG ; Gao-Sheng HU ; Jing-Ming JIA
Chinese Journal of Natural Medicines (English Ed.) 2022;20(7):527-536
Phenylalanine ammonia-lyase (PAL), which catalyzes the conversion from L-phenylalanine to trans-cinnamic acid, is a well-known key enzyme and a connecting step between primary and secondary metabolisms in the phenylpropanoid biosynthetic pathway of plants and microbes. Schisandra chinensis, a woody vine plant belonging to the family of Magnoliaceae, is a rich source of dibenzocyclooctadiene lignans exhibiting potent activity. However, the functional role of PAL in the biosynthesis of lignan is relatively limited, compared with those in lignin and flavonoids biosynthesis. Therefore, it is essential to clone and characterize the PAL genes from this valuable medicinal plant. In this study, molecular cloning and characterization of three PAL genes (ScPAL1-3) from S. chinensis was carried out. ScPALs were cloned using RACE PCR. The sequence analysis of the three ScPALs was carried out to give basic characteristics followed by docking analysis. In order to determine their catalytic activity, recombinant protein was obtained by heterologous expression in pCold-TF vector in Escherichia coli (BL21-DE3), followed by Ni-affinity purification. The catalytic product of the purified recombinant proteins was verified using RP-HPLC through comparing with standard compounds. The optimal temperature, pH value and effects of different metal ions were determined. Vmax, Kcat and Km values were determined under the optimal conditions. The expression of three ScPALs in different tissues was also determined. Our work provided essential information for the function of ScPALs.
Cloning, Molecular
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Escherichia coli/metabolism*
;
Phenylalanine/metabolism*
;
Phenylalanine Ammonia-Lyase/chemistry*
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Recombinant Proteins
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Schisandra/genetics*
3.Pathogenesis of Esophageal Cancer and Clinical Intervention of Traditional Chinese Medicine: A Review
Meng-fan PENG ; Shuo TIAN ; Chen-hui LI ; Yuan-xin ZHANG ; Jin-xin MIAO ; Ming-san MIAO
Chinese Journal of Experimental Traditional Medical Formulae 2022;28(12):267-274
Esophageal cancer is a digestive tract malignancy with high morbidity and mortality and mainly occurs in males. The 5-year survival rate is lower than 20%. In China, the morbidity and mortality of esophageal cancer rank the first in the world, seriously threatening national health. The pathogenesis of esophageal cancer is diverse, which is generally considered as the consequence of environmental-genetic-gene interaction. In addition to genetic factors and regional characteristics, gene mutation, RNA interference, DNA damage repair, tumor microenvironment, dietary habit, chronic adverse stimulation, and inflammatory reaction are all involved in the occurrence and development of esophageal cancer. However, there is no unified and accurate conclusion. Clarifying the exact pathogenesis of esophageal cancer is of great significance for its early screening, diagnosis, prevention, treatment, and prognosis. Surgery, radiotherapy, and chemotherapy are the three effective methods for the treatment of esophageal cancer. However, due to the atypical early symptoms, most patients have missed the best operation period when diagnosed, resulting in poor clinical prognosis. Moreover, radiotherapy and chemotherapy will cause side effects such as loss of appetite, low immune function, esophagitis, pneumonia, and malnutrition, which is not conducive to the prognosis and treatment maintenance of patients. With definite efficacies on esophageal cancer, traditional Chinese medicine (TCM), which is flexible and diverse in the treatment, can primarily or alternatively be involved in the treatment of esophageal cancer. TCM can eliminate postoperative complications and postoperative infections and relieve adverse gastrointestinal reactions, weakened immune function, and organ damage caused by radiotherapy and chemotherapy. It can enhance clinical efficacy and improve the quality of life of patients. Therefore, it is necessary to systematically summarize the clear pathogenesis or risk factors of esophageal cancer and review the clinical characteristics of TCM in the prevention and treatment of esophageal cancer to facilitate the early screening, diagnosis, and treatment of esophageal cancer and promote the application of TCM in the prevention and treatment of esophageal cancer and related adverse reactions.
4.A new norsesquiterpenoid from Schisandra chinensis.
Yong-Cheng YANG ; Jiang-Chun WEI ; Lu-Qi LIU ; San-Peng FAN ; Min CHEN ; An-Hua WANG ; Jing-Ming JIA
China Journal of Chinese Materia Medica 2022;47(6):1582-1586
This study investigated the chemical components from the leaves and stems of Schisandra chinensis. Three norsesquiterpenoids were isolated from S. chinensis by various column chromatographies(silica gel, Sephadex LH-20, and MCI), reversed-phase medium-pressure preparative, and semi-preparative high-performance liquid chromatography(HPLC). Their structures were identified based on physicochemical properties, mass spectrometry(MS), nuclear magnetic resonance(NMR), ultraviolet(UV), and electro-nic circular dichroism(ECD) as(3R,4R,5R,6S,7E)-3,4,5,6-tetrahydroxy-7-megastigmen-9-one(1),(3S,5R,6R,7E)-3,5,6-trihydroxy-7-megastigmen-9-one(2), and(3S,4R,9R)-3,4,9-trihydroxymegastigman-5-ene(3). Compound 1 was a new compound, and its absolute configuration was determined by ECD. Compounds 2 and 3 were isolated from the Schisandra plant for the first time.
Chromatography, High Pressure Liquid
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Magnetic Resonance Spectroscopy
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Mass Spectrometry
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Plant Leaves/chemistry*
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Schisandra
5.Research progress of Phyllanthi Fructus and prediction of its Q-markers.
Hao-Zhou HUANG ; Jing-Cai CHEN ; Ding-Kun ZHANG ; Meng-Qi LI ; Qin-Chi XIAN ; San-Hu FAN ; Peng TAN ; Wan-Min MAO ; Feng LIN ; Jun-Zhi LIN ; Li HAN
China Journal of Chinese Materia Medica 2021;46(21):5533-5544
Phyllanthi Fructus, a unique Chinese and Tibetan medicinal plant with both edible and medical values, has high potential of cultivation and development. The resources of Phyllanthi Fructus in China are rich, mainly distributed in Yunnan, Sichuan, Fujian, Guangdong, Guangxi, etc. Phyllanthi Fructus is widely used in the clinical practice of Chinese medicine and plays an important role in Tibetan medicine, Uyghur medicine, Yi medicine, and Mongolian medicine. Phyllanthi Fructus mainly contains phenolic acids,tannins, terpenes, sterols, fatty acids, flavonoids, amino acids and other compounds. Modern pharmacological studies show that Phyllanthi Fructus has antioxidant, anticancer, blood lipid-lowering, liver protective, antimicrobial, anti-inflammatory, and immune regulatory activities. In this paper, the research status of Phyllanthi Fructus was reviewed from the aspects of herbal textual research,chemical composition, and pharmacological action. The quality markers(Q-markers) of Phyllanthi Fructus were predicted and analyzed from the aspects of biogenic pathway, specificity and measurability of chemical components, efficacy, properties, new clinical uses, drug-food homology, and transformation of polyphenols. The results will provide a scientific basis for the quality control, quality evaluation, and standard formulation of Phyllanthi Fructus.
China
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Drugs, Chinese Herbal
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Fruit
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Medicine, Tibetan Traditional
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Quality Control
6.Effect of Endoplasmic Reticulum Stress on Tumorigenesis and Development and Intervention of Traditional Chinese Medicine: A Review
Meng-fan PENG ; Jin-xin MIAO ; Shuo TIAN ; Xue WANG ; Hai-tao LU ; Ming-san MIAO
Chinese Journal of Experimental Traditional Medical Formulae 2021;27(22):215-221
Endoplasmic reticulum (ER) is an important organelle responsible for protein, steroid, lipid and carbohydrate synthesis and calcium-dependent signal transduction in eukaryotic cells. ER homeostasis is essential for normal cell function. ER homeostasis imbalance can induce ER stress (ERS), which participates in the occurrence and development of diseases of the digestive system, respiratory system, circulatory system, nervous system, reproductive system, and endocrine system, and affects body health. Among various diseases, cancers seriously endanger people′s health due to its high mortality rate, disability rate, and recurrence rate. Due to the survival characteristics of unlimited proliferation, tumor cells are often exposed to various internal and external stimuli such as hypoxia, ischemia, excessive proliferation, and starvation, which destroy intracellular protein balance and induce ERS to some extent for survival. ERS plays a major role in various tumors and has dual functions in the survival of tumor cells: promoting the survival of tumor cells by activating a series of adaptive responses, while inducing ERS-related apoptosis pathways, so as to promote tumor cell death and inhibit tumor growth and invasion. As multiple functions of ERS in tumors are reported, many scholars have tried to intervene in the progress of tumors from the perspective of ERS. The therapeutic effect of traditional Chinese medicine (TCM) on tumors has been widely recognized. TCM can participate in the regulation of tumors from many aspects, including ERS, chemoradiotherapy resistance, gastrointestinal adverse reactions caused by chemotherapy, postoperative recurrence and metastasis. Since there are few reports on the antitumor effect of TCM from the perspective of ERS, this paper expounds the influence of ERS on tumorigenesis and development and the progress of TCM intervention in tumor through ERS, in order to provide a new direction for tumor treatment.
7.Effect of Removing Core on Quality of Phyllanthi Fructus
Chuan-hong LUO ; Sheng-jie HUANG ; Qi-qi HU ; Peng TAN ; Hua-mei GOU ; Xi-chuan WEI ; Hao-zhou HUANG ; San-hu FAN ; Jun-zhi LIN ; Ding-kun ZHANG ; Li HAN
Chinese Journal of Experimental Traditional Medical Formulae 2021;27(9):147-156
Objective:The differences of chemical compositions and pharmacological activities between the core and pulp of Phyllanthi Fructus were investigated by chemical analysis and
8.Discussion on
Chang-Zhen GONG ; Fan-Rong LIANG ; Can-Hui LI ; Wei-Xing PAN ; Yong-Ming LI ; San-Hua LENG ; Arthur Yin FAN ; Song-Ping HAN ; Jing LIU ; Shan WANG ; Zeng-Fu PENG ; Ye-Meng CHEN ; Guan-Hu YANG ; Xu-Ming GU ; Hong SU ; Shao-Bai WANG
Chinese Acupuncture & Moxibustion 2021;41(4):359-364
Professor
Acupuncture
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Acupuncture Therapy
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Angina, Stable
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Combined Modality Therapy
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Humans
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Moxibustion
9.Strategies and ideas of comprehensive development and utilization of medicine and food homologous variety Phyllanthus emblica.
Hao-Zhou HUANG ; Fei RAN ; Qing-Chu TAN ; Ding-Kun ZHANG ; Meng-Qi LI ; San-Hu FAN ; Peng TAN ; Jin PEI ; Li HAN ; Jun-Zhi LIN
China Journal of Chinese Materia Medica 2021;46(5):1034-1042
Phyllanthus emblica is a kind of traditional medicine and medicinal and edible plant, with rich variety resources and high development value. It is a key poverty alleviation variety in China at present. As P. emblica processing industry is rising gradually in recent years, in order to fully develop and utilize its industrial resources, this paper systematically introduces current comprehensive development and utilization of P. emblica, discusses the problems in P. emblica processing industry, and puts forward comprehensive development and utilization strategies and industrial models in terms of cultivation, breeding, grading, quality evaluation and waste recycling, so as to provide a certain reference for promoting the high-quality development of P. emblica industry in China.
China
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Medicine
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Medicine, Traditional
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Phyllanthus emblica
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Plant Breeding
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Plant Extracts
10.Analysis of Chemical Components in Supernatant and Precipitate of Sanajon Oral Liquid Based on UPLC-Q-TOF-MSE Technology
Hao-zhou HUANG ; Min QIU ; Jin-fa TANG ; Meng-Qi LI ; Hai-yan LIU ; San-hu FAN ; Peng TAN ; Li HAN ; Jun-zhi LIN ; Ding-kun ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2020;26(14):143-151
Objective:To analyze the main chemical components in the supernatant and precipitate of Sanajon oral liquid, so as to provide basis for establishing its quality standard and precipitation control technology. Method:UPLC-Q-TOF-MSE was used to analyze the chemical components in the supernatant and precipitate of this oral liquid. The analysis was performed on Waters ACQUITY UPLC BEH C18 column (2.1 mm×100 mm, 1.7 μm) with the mobile phase of 0.1% formic acid solution (A) and acetonitrile (B) for gradient elution (0-1 min, 2%B; 1-2 min, 2%-5%B; 2-4 min, 5%-7%B; 4-6 min, 7%-24%B; 6-10 min, 24%-42%B; 10-12 min, 42%-54%B; 12-15 min, 54%-76%B; 15-18 min, 76%-100%B), the flow rate was set to 0.3 mL·min-1, the column temperature was 30 ℃, the injection volume was 2 µL. The mass spectrographic analysis was used with electrospray ionization (ESI), sample MS data was acquired by time-dependent MSE in negative ion mode, the collection range was

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