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Overexpression of TRPV1 activates autophagy in human lens epithelial cells under hyperosmotic stress through Ca^(2+)-dependent AMPK/mTOR pathway

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International Journal of Ophthalmology(English edition) 2024年第3期17卷 420-434页

作者： Liu-Hui Huang Jiao Lyu Sheng Chen Ting-Yi Liang Yu-Qing Rao Ping Fei Jing Li Hai-ying Jin Pei-Quan Zhao Department of Ophthalmology Shanghai Xinhua Hospital Affiliated with Shanghai Jiao Tong University School of MedicineShanghai 200092China Department of Ophthalmology Shanghai Tenth People’s Hospital Affiliated with Shanghai Tongji University School of MedicineShanghai 200072China Centre for Eye Research Australia Royal Victorian Eye and Ear HospitalEast MelbourneVictoria 3002Australia Ophthalmology Department of SurgeryUniversity of MelbourneEast MelbourneVictoria 3002Australia Shenzhen Eye Hospital Shenzhen Key Laboratory of OphthalmologyAffiliated Shenzhen Eye Hospital of Jinan UniversityShenzhen 518040Guangdong ProvinceChina Department of Ophthalmology Shanghai East HospitalSchool of MedicineTongji UniversityShanghai 200120China

●AIM:To explore whether autophagy functions as a cellular adaptation mechanism in lens epithelial cells(LECs)under hyperosmotic stress.●METHODS:LECs were treated with hyperosmotic stress at the concentration of 270,300,400,500,or 600 mOsm for 6,12,18,24h in *** chain reaction(PCR)was employed for the mRNA expression of autophagyrelated genes,while Western blotting detected the targeted protein *** transfection of stub-RFP-sens-GFPLC3 autophagy-related double fluorescence lentivirus was conducted to detect the level of autophagy *** electron microscopy was used to detect the existence of *** interfering RNA of autophagy-related gene(ATG)7,transient receptor potential vanilloid(TRPV)1 overexpression plasmid,related agonists and inhibitors were employed to their influence on autophagy related *** cytometry was employed to test the apoptosis and intracellular Ca^(2+)*** membrane potential was measured by JC-1 *** cell counting kit-8 assay was used to calculate the cellular *** wound healing assay was used to evaluate the wound closure *** 6.0 software was utilized to evaluate the data.●RESULTS:The hyperosmotic stress activated autophagy in a pressure-and time-dependent manner in *** 1 protein expression and conversion of LC3B II to LC3B I increased,whereas sequestosome-1(SQSTM1)protein expression *** Ca^(2+)influx was stimulated caused by hyperosmotic stress,levels of mammalian target of rapamycin(mTOR)phosphorylation decreased,and the level of AMP-activated protein kinase(AMPK)phosphorylation increased in the early *** on this evidence,autophagy activation through the Ca^(2+)-dependent AMPK/mTOR pathway might represent an adaptation process in LECs under hyperosmotic *** stress decreased cellular viability and accelerated apoptosis in LECs and cellular migration *** of autophagy by ATG7 knockdown had sim

关键词： cataract posterior capsular opacification lens epithelial cell hyperosmotic stress autophagy apoptosis transient receptor potential vanilloid 1

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Development of an Integrated Disposable Device for SARSCoV-2 Nucleic Acid Extraction and Detection

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Biomedical and Environmental Sciences 2024年第6期37卷 639-646页

作者： Jing Ma Yanzhe Hao Meiling Hou Xiaoshan Zhang Jingduan Liu Haodi Meng Jiangbo Chang Xuejun Ma Jihua Liu Qingjie ying Xianhua Wang Hongxia Li Yuxi Cao Xiaoguang Zhang National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases National Institute for Viral Disease Control and PreventionChinese Center for Disease Control and PreventionBeijing 102206China Beijing Runbio Biotechnology Development Co. LtdBeijing 100106China Beijing Beta Technology Co. LtdBeijing 101318China Jiangsu Qitian Gene Biotechnology Co. LtdWuxi 214135JiangsuChina

Objective To develop a highly sensitive and rapid nucleic acid detection method for the severe acute respiratory syndrome coronavirus 2(SARS-CoV-2).Methods We designed,developed,and manufactured an integrated disposable device for SARS-CoV-2 nucleic acid extraction and *** precision of the liquid transfer and temperature control was tested.A comparison between our device and a commercial kit for SARS-Cov-2 nucleic acid extraction was performed using real-time fluorescence reverse transcription polymerase chain reaction(RT-PCR).The entire process,from SARS-CoV-2 nucleic acid extraction to amplification,was *** The precision of the syringe transfer volume was 19.2±1.9μL(set value was 20),32.2±1.6(set value was 30),and 57.2±3.5(set value was 60).Temperature control in the amplification tube was measured at 60.0±0.0℃(set value was 60)and 95.1±0.2℃(set value was 95)***-Cov-2 nucleic acid extraction yield through the device was 7.10×10^(6) copies/mL,while a commercial kit yielded 2.98×10^(6) copies/*** mean time to complete the entire assay,from SARS-CoV-2 nucleic acid extraction to amplification detection,was 36 min and 45 *** detection limit for SARS-CoV-2 nucleic acid was 250 copies/*** The integrated disposable devices may be used for SARS-CoV-2 Point-of-Care test(POCT).

关键词： An integrated disposable device SARS-Cov-2 Nucleic acid detection

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Method for the extraction of circulating nucleic acids based on MOF reveals cell-free RNA signatures in liver cancer

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National Science Review 2024年第1期11卷 94-106页

作者： Yuqing Sun Haixin Yu Shaoqing Han Ruoxi Ran ying Yang Yongling Tang Yuhao Wang Wenhao Zhang Heng Tang Boqiao Fu Boshi Fu Xiaocheng Weng Song-Mei Liu Hexiang Deng Shuang Peng Xiang Zhou College of Chemistry and Molecular Sciences Key Laboratory of Biomedical Polymers-Ministry of Education Wuhan University Department of Digestive Surgical Oncology Union Hospital Tongji Medical College Huazhong University of Science and Technology Cancer Center Union HospitalTongji Medical CollegeHuazhong University of Science and Technology Department of Clinical Laboratory Center for Gene Diagnosis and Program of Clinical Laboratory Zhongnan Hospital of Wuhan University College of Chemistry and Materials Science Hubei Engineering University Department of Pharmacology School of Pharmacy China Medical University Taikang Center for Life and Medical Sciences Wuhan University Department of Hematology Zhongnan Hospital of Wuhan University

Cell-free RNA(cf RNA) allows assessment of health, status, and phenotype of a variety of human organs and is a potential biomarker to non-invasively diagnose numerous diseases. Nevertheless, there is a lack of highly efficient and bias-free cf RNA isolation technologies due to the low abundance and instability of cf ***, we developed a reproducible and high-efficiency isolation technology for different types of cell-free nucleic acids(containing cf RNA and viral RNA) in serum/plasma based on the inclusion of nucleic acids by metal-organic framework(MOF) materials, which greatly improved the isolation efficiency and was able to preserve RNA integrity compared with the most widely used research kit method. Importantly, the quality of cf RNA extracted by the MOF method is about 10-fold that of the kit method, and the MOF method isolates more than three times as many different RNA types as the kit method. The whole transcriptome mapping characteristics of cf RNA in serum from patients with liver cancer was described and a cf RNA signature with six cf RNAs was identified to diagnose liver cancer with high diagnostic efficiency(area under curve = 0.905 in the independent validation cohort) using this MOF method. Thus, this new MOF isolation technique will advance the field of liquid biopsy, with the potential to diagnose liver cancer.

关键词： MOF circulating cell-free RNA biomarker liver cancer diagnosis

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Urocortin and cardiovascular protection

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Acta Pharmacologica Sinica 2004年第3期25卷 3-11页

作者： Chi-wai LAU,Yau-chi CHAN,Suk-ying TSANG,Franky leung CHAN The Chinese University of Hong Kong Hong Kong China Department of Anatomy The Chinese University of Hong Kong Hong Kong China Department of Anatomy The Chinese University of Hong Kong Hong Kong China Department of Anatomy The Chinese University of Hong Kong Hong Kong China Department of Anatomy

Urocortin and other hypothalamus corticotropin-releasing factor (CRF) polypeptides play biologically diverse roles in the stress, cardiovascular and inflammatory responses by acting on central and peripheral CRF receptors. Urocortin shows a significantly high sequence homology to CRF, and the concurrent expression of type-2 CRF (CRF2) receptors with urocortin in the heart suggests that urocortin may play a physiological role in the cardiac function. Urocortin is thought to be the endogenous agonist producing the cardiovascular actions previously attributed to CRF. This review highlights the current novel findings on the molecular and cellular mechanisms by which urocortin may exert its cardiovascular protective action.

关键词： urocortin corticotropin-releasing hormone vascular smooth muscle heart vasodilaion second messenger system protein kinases

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Experimental study on mechanical behaviors of pseudo-ductile cementitious composites under biaxial compression

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Science China(Technological Sciences) 2013年第4期56卷 963-969页

作者： ZHOU JiaJia PAN JinLong leung Christopher Kin ying LI ZongJin Key Laboratory of Concrete and Prestressed Concrete Structures of Ministry of Education Southeast University Department of Civil and Environmental Engineering Hong Kong University of Science and Technology

Experimental investigation was conducted to characterize the responses of pseudo-ductile cementitious composites (PDCCs) when subjected to uniaxial and biaxial *** PDCCs is a class of fiber reinforced cementitious composites with ultra-high ductility by using a low volume fraction (2%) of polyvinyl alcohol (PVA) *** different strength grades of PDCC were examined with cubic specimen size of 100 mm in the *** specimens were loaded with a servo-hydraulic jack at different stress *** principle stresses and strains of the specimens were recorded,and the failure modes with various stress states were *** test results indicated that the ultimate strength of PDCCs increased due to the lateral confinement in the other principal stress direction,and the maximum ultimate strength occurred at the biaxial stress ratio of 0.25,which was very different from common concrete *** the PDCC specimens,the biaxial strength may be lower than the uniaxial strength when subjected to biaxial compression with the stress ratio of 1.0,and the failure mode showed a shear-type failure because of the bridging effect of ***,a failure criterion was proposed for PDCCs under biaxial compression.

关键词： PDCC biaxial strength stress-strain curve failure mode failure criterion

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cNPAS2 inducedβcell dysfunction by regulating KANK1 expression in type 2 diabetes

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World Journal of Diabetes 2024年第9期15卷 1932-1941页

作者： Yan-Bin Yin Wei Ji ying-Lan Liu Qian-Hao Gao Dong-Dong He Shi-Lin Xu Jing-Xin Fan Li-Hai Zhang Department of General Surgery The First Affiliated Hospital of Jiamusi UniversityJiamusi 154000Heilongjiang ProvinceChina Department of Anesthesiology Yantai Affiliated Hospital of Binzhou Medical UniversityYantai 264199Shandong ProvinceChina Operating Room The First Affiliated Hospital of Jiamusi UniversityJiamusi 154000Heilongjiang ProvinceChina Department of Anesthesiology Huazhong University of Science and Technology Union Jiangbei HospitalWuhan 430100Hubei ProvinceChina Department of Endocrinology The First Affiliated Hospital of Jiamusi UniversityJiamusi 154000Heilongjiang ProvinceChina

BACKGROUND Diabetes mellitus type 2(T2DM)is formed by defective insulin secretion with the addition of peripheral tissue resistance of insulin *** has been affecting over 400 million people all over the *** To explore the pathogenesis of T2DM and to develop and implement new prevention and treatment strategies for *** Receiver operating characteristic(ROC)curve analysis was used to conduct diagnostic *** expression level of genes was determined by reverse transcription-PCR as well as Western *** proliferation assays were performed by cell counting kit-8(CCK-8)*** last,T2DM mice underwent Roux-en-Y gastric bypass *** We found that NPAS2 was significantly up-regulated in isletβcell apoptosis of *** ROC curve revealed that NPAS2 was capable of accurately diagnosing ***2 overexpression did increase the level of *** addition,the CCK-8 test revealed knocking down NPAS2 and KANK1 increased the proliferation of MIN6 *** last,we found that gastric bypass may treat type 2 diabetes by down-regulating NPAS2 and *** This study demonstrated that NPAS2 inducedβcell dysfunction by regulating KANK1 expression in type 2 diabetes,and it may be an underlying therapy target of T2DM.

关键词： Diabetes mellitus type 2 KANK1 NPAS2 Gastric bypass

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Screening and identification of mimotopes of the major shrimp allergen tropomyosin using one-bead-one- compound peptide libraries

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Cellular & Molecular Immunology 2017年第3期14卷 308-318页

作者： Nicki YH leung Christine YY Wai Marco HK Ho Ruiwu Liu kit S Lam Jin Jun Wang Shang An Shu Ka Hou Chu Patrick SC leung School of Life Sciences The Chinese University of Hong Kong Shatin N.T. Hong Kong SAR China Department of Pediatrics and Adolescent Medicine Queen Mary Hospital The University of Heng Kong Pokfulam Hong Kong SAR China Department of Biochemistry and Molecular Medicine University of California Davis CA 95616 USA Division of Rheumatology/Allergy School of Medicine University of California Davis CA 95616 USA

The one-bead-one-compound （OBOC） combinatorial peptide library is a powerful tool to identify ligand and receptor interactions. Here, we applied the OBOC library technology to identify mimotopes specific to the immunoglobulin E （IgE） epitopes of the major shellfish allergen tropomyosin. OBOC peptide libraries with 8-12 amino acid residues were screened with serum samples from patients with shellfish allergy for IgE mimotopes of tropomyosin. Twenty-five mimotopes were identified from the screening and their binding reactivity to tropomyosin-specific IgE was confirmed by peptide ELISA. These mimotopes could be divided into seven clusters based on sequence homology, and epitope mapping by EpiSearch of the clustered mimotopes was performed to characterize and confirm the validity of mimotopes. Five out of six of the predicted epitopes were found to overlap with previously identified epitopes of tropomyosin. To further confirm the mimicry potential of mimotopes, BALB/c mice were immunized with mimotopes conjugated to keyhole limpet hemocyanin and assayed for their capacity to induce tropomyosin-specific antibodies. BALB/c mice that received mimotope immunization were found to have an elevated level of tropomyosin-specific immunoglobulin G, but not mice that received an irrelevant mimotope. This study pioneers the successful application of the OBOC libraries using whole sera to screen and identify multiple shrimp allergen mimotopes and validates their mimicry potential using in vitro, in vivo, and in silico methods.

关键词： epitope mimotope OBOC peptide library tropomyosin

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Potential application of Nardostachyos Radix et Rhizoma-Rhubarb for the treatment of diabetic kidney disease based on network pharmacology and cell culture experimental verification

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World Journal of Diabetes 2024年第3期15卷 530-551页

作者： Meng-ying Che Ling Yuan Jiao Min Duo-Jie Xu Dou-Dou Lu Wen-Jing Liu Kai-Li Wang Yan-Yan Wang Yi Nan Key Laboratory of Ningxia Minority Medicine Modernization Ministry of Education Ningxia Medical UniversityYinchuan 750004Ningxia Hui Autonomous RegionChina College of Pharmacy Ningxia Medical UniversityYinchuan 750004Ningxia Hui Autonomous RegionChina Traditional Chinese Medicine College Ningxia Medical UniversityYinchuan 750004Ningxia Hui Autonomous RegionChina School of Clinical Medicine Ningxia Medical UniversityYinchuan 750004Ningxia Hui Autonomous RegionChina Department of Endocrinology Yinchuan Hospital of Traditional Chinese MedicineYinchuan 750004Ningxia Hui Autonomous RegionChina

BACKGROUND Diabetic kidney disease(DKD)is one of the serious complications of diabetes mellitus,and the existing treatments cannot meet the needs of today's *** Chinese medicine has been validated for its efficacy in DKD after many years of clinical ***,the specific mechanism by which it works is still *** the molecular mechanism of the Nardostachyos Radix et Rhizoma-rhubarb drug pair(NRDP)for the treatment of DKD will provide a new way of thinking for the research and development of new *** To investigate the mechanism of the NRDP in DKD by network pharmacology combined with molecular docking,and then verify the initial findings by in vitro *** The Traditional Chinese Medicine Systems Pharmacology(TCMSP)database was used to screen active ingredient targets of *** for DKD were obtained based on the Genecards,OMIM,and TTD *** VENNY 2.1 database was used to obtain DKD and NRDP intersection targets and their Venn diagram,and Cytoscape 3.9.0 was used to build a"drug-component-target-disease"*** String database was used to construct protein interaction *** Encyclopedia of Genes and Genomes(KEGG)enrichment analysis and Gene Ontology analysis were performed based on the DAVID *** selecting the targets and the active ingredients,Autodock software was used to perform molecular *** experimental validation using renal tubular epithelial cells(TCMK-1),we used the Cell Counting kit-8 assay to detect the effect of NRDP on cell viability,with glucose solution used to mimic a hyperglycemic *** cytometry was used to detect the cell cycle progression and *** blot was used to detect the protein expression of STAT3,p-STAT3,BAX,BCL-2,Caspase9,and *** A total of 10 active ingredients and 85 targets with 111 disease-related signaling pathways were obtained for *** analysis of KEGG pathways was performed to determ

关键词： Nardostachyos Radix et Rhizoma-rhubarb Diabetic kidney disease Molecular docking Network pharmacology Experimental validation

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Contents of four active components in different commercial crude drugs and preparations of Danshen (Salvia miltiorrhiza)

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Acta Pharmacologica Sinica 2002年第12期23卷 110-115页

作者： Susan Wai Sum leung,Yaw Loong SIOW,Ricky ying Keung MAN Department of Pharmacology Faculty of Medicine University of Hong Kong 21 Sassoon Road Hong Kong China Department of Pharmacology Faculty of Medicine University of Hong Kong 21 Sassoon Road Hong Kong China Department of Pharmacology Faculty of Medicine University of Hong Kong 21 Sassoon Road Hong Kong China Department of Pharmacology University of Hong Kong 21 Sassoon Road Hong Kong China

AIM: To detect the contents of four active components of Salvia miltiorrhiza in various commercially available danshen crude drugs and preparations. METHODS: Commercially available danshen crude drugs from different sources, as well as danshen pills and intravenous injection preparations containing danshen alone or in combination with other herbs were collected. The composition of these danshen samples was analyzed using HPLC. Specifically, the amounts of magnesium tanshinoate B (MTB), danshensu, isotanshinone IIA, and ctyptotanshinone were determined. In some of these samples, the content of MTB was further confirmed by liquid chromatography-tandem mass spectrometer (LC-MS)/MS method. RESULTS: There were great variations in the amount of the four active ingredients in the commercially available danshen crude drugs and drug preparations in this study. The amount of MTB was the highest among the four components measured in the crude drugs. However, the amounts of MTB in all danshen preparations were much

关键词： Salvia miltiorrhiza magnesium tanshinoate B (MTB) high pressure liquid chromatography

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Mechanism of metastasis by membrane type 1-matrix metalloproteinase in hepatocellular carcinoma

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World Journal of Gastroenterology 2005年第40期11卷 6269-6276页

作者： ying-Chi Ip Siu-Tim Cheung Ka-Ling leung Sheung-Tat Fan Centre for the Study of Liver Disease and Department of Surgery

AIM To investigate the precise role of membrane type 1-matrix metalloproteinase （NTI-NNP） in hepatocellular carcinoma （HCC） metastasis. METHODS- Human HCC cells Hep3B with overexpression of MTT-MMP were established by stable transfection, and compared with control cells carrying the empty vector. Cells were examined in vivo for their differences in the metastatic ability of athymic nude mice, and analyzed in vito for their differences in invasion ability by invasion chamber coated with Matrigel, adhesion towards collagen I and migration through culture chamber. Cell proliferation and apoptosis in adherent and suspension status were evaluated by MTr and flow cytometry analysis. RESULTS We found that overexpression of MT1-MMP could increase intrahepatic metastasis in nude mice with orthotopic implantation of HCC cells （incidence of 100% [MT1-MMP transfectants] vs 40% [vector control transfectants], P〈0.05）. NT1-MMP could also enhance cell invasion through Natrigel （107.7 vs 39.3 cells/field, P〈0.001）, adhesion towards matrix （0.30 vs 0.12 absorbance unit at 540 nm, P〈0.001）, cell migration （89.3 vs 39.0 cells/field, P〈0.001）, and cell proliferation （24.3 vs 40.5 h/doubling, P〈0.001）. We also observed that NTI-NNP supported cell survival （71.4% vs 23.9%, P〈0.001） with reduced apoptosis （43.7% vs51.0%, P〈0.05） in an attachment-free environment. CONCLUSION： MT1-MMP overexpression could enhance metastasis. In addition to its active role in matrix degradation during tumor invasion, MT1-MMP enhances tumor cell survival upon challenge of detachment, which is important during metastasis when cells enter the circulation.

关键词： MT1-MMPIMMP14 Liver cancer Invasion Metastasis

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