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Expression of EGFP/SDCT1 fusion protein,subcellular localization signal analysis,tissue distribution and electrophysiological function study

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Science China(Life Sciences) 2004年第6期47卷 530-539页

作者： BAI Xueyuan CHEN Xiangmei FEN Zhe WU Di HOU Kai CHENG Genyang PENG Lixia Chinese PLA kidney Center & Key Laboratory for Nephrology Chinese PLA General Hospital & Military Medical Postgraduate CollegeBeijing 100853China

Full-length cDNA gene of sodium-dependent dicarboxylate co-transporter protein 1 (SDCT1) is cloned from normal human kidney tissue and inserted into EGFP (enhanced green fluorescent protein) expression vector along with N-terminal and C-terminal truncated SDCT1 genes, so to construct the eukaryotic expression vectors of EGFP/SDCT1 fusion proteins, which are transfected into human renal tubular epithelial cells (HKC). Subcellular localizations of these fusion proteins are observed by laser confocal microscope to determine the localization signal of the SDCT1 protein. Duplex PCR analysis validates that the fusion protein genes have been in- tegrated into the genome of HKC. Western blot indicates that the fusion proteins have been ex- pressed in HKC. Confocal microscopy analysis shows that human SDCT1 predominantly locates on the plasma membrane, which is consistent with the results predicted by bioinformatics ap- proach; in HKC transfected with N-terminal truncated SDCT1 gene, the green fluorescence is mainly distributed on the plasma membrane; in HKC transfected with C-terminal truncated SDCT1 gene, the green fluorescence is mainly distributed in the cytoplasm. EGFP/SDCT1 mRNAs obtained by in vitro transcription are microinjected into Xenopus laevis oocytes for ex- pression and the trans-membrane currents are measured by using two-microelectrode volt- age-clamp technique. Na+ inward currents are present on cellular membrane of the injected oo- cytes. Immunohistochemical staining shows that human SDCT1 proteins are expressed on lu- men membrane of the renal proximal tubule, but are negative in distal tubule, collecting duct, renal interstitium and glomerulus. The above-mentioned studies suggest that human SDCT1 protein is located on the lumen membrane of the renal proximal tubule, the C-terminal sequence of the SDCT1 is required for delivery and targeting localization, and the plasma membrane lo- calization signal of the SDCT1 protein maybe locate in the C-terminal se

关键词： dicarboxylate co-transporter tricarboxylic acid cycle renal tubule subcellular localization.

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Chronic heat stress induces renal fibrosis and mitochondrial dysfunction in laying hens

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畜牧与生物技术杂志:英文版 2023年第5期14卷 2002-2011页

作者： Fumika Nanto‑Hara Makoto Yamazaki Hitoshi Murakami Haruhiko Ohtsu Division of Meat Animal and Poultry research Institute of Livestock and Grassland ScienceNational Agriculture and Food Research Organization(NILGS)2 IkenodaiTsukubaIbaraki 305‑0901Japan

Background Heat stress in laying hens negatively affects egg production and shell quality by disrupting the homeo-stasis of plasma calcium and phosphorus *** the kidney plays an important role in calcium and phos-phorus homeostasis,evidence regarding the effect of heat stress on renal injury in laying hens is yet to be ***,the aim of this study was to evaluate the effects of chronic heat stress on renal damage in hens during laying *** A total of 16 white-leghorn laying hens(32 weeks old)were randomly assigned to two groups(n=8).One group was exposed to chronic heat stress(33°C for 4 weeks),whereas the other group was maintained at 24°*** Chronic heat exposure significantly increased plasma creatinine and decreased plasma albumin levels(P<0.05).Heat exposure also increased renal fibrosis and the transcription levels of fibrosis-related genes(COLA1A1,αSMA,and TGF-β)in the *** results suggest that renal failure and fibrosis were induced by chronic heat exposure in laying *** addition,chronic heat exposure decreased ATP levels and mitochondrial DNA copy number(mtDNA-CN)in renal tissue,suggesting that renal mitochondrial dysfunction occurs under conditions of heat *** mitochondria leak mtDNAs into the cytosol and mtDNA leakage may activate the cyclic GMP-AMP synthase(cGAS)stimulator of interferon genes(STING)signaling *** results showed that chronic heat exposure activated the cGAS-STING pathway as indicated by increased expression of MDA5,STING,IRF7,MAVS,and NF-κB ***,the expression of pro-inflammatory cytokines(IL-12)and chemokines(CCL4 and CCL20)was upregulated in heat-stressed *** These results suggest that chronic heat exposure induces renal fibrosis and mitochondrial damage in laying *** damage by heat stress may activate the mtDNA-cGAS-STING signaling and cause subse-quent inflammation,which contributes to the progression of renal fibrosis and

关键词： cGAS-STING Heat stress Laying hens Mitochondrial DNA Renal fibrosis

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Predictive Score Model for Delayed Graft Function Based on Hypothermic Machine Perfusion Variables in kidney Transplantation

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Chinese Medical Journal 2018年第22期131卷 2651-2657页

作者： Chen-Guang Ding Yang Li Xiao-Hui Tian Xiao-Jun Hu Pu-Xun Tian Xiao-Ming Ding He-Li Xiang Jin Zheng Wu-Jun Xue Department of kidney Transplantation Nephropathy HospitalThe First Affiliated Hospital of Xi'an Jiaotong UniversityXi'anShaanxi 710061China Institute of Organ Transplantation Xi'an Jiaotong UniversityXi'anShaanxi 710061China

Background: Hypothermic machine perfusion(HMP) is being used more often in cardiac death kidney transplantation; however, the significance of assessing organ quality and predicting delayed graft function(DGF) by HMP parameters is still controversial. Therefore,we used a readily available HMP variable to design a scoring model that can identify the highest risk of DGF and provide the guidance and advice for organ allocation and DCD kidney ***: From September 1, 2012 to August 31, 2016, 366 qualified kidneys were randomly assigned to the development and validation cohorts in a 2:1 distribution. The HMP variables of the development cohort served as candidate univariate predictors for DGF. The independent predictors of DGF were identified by multivariate logistic regression analysis with a P kidney. The validation cohort was used to verify the accuracy and reliability of the scoring ***: HMP duration(OR = 1.165, 95% confidence interval [CI ]: 1.008–1.360, P = 0.043), resistance(OR = 2.190, 95%CI: 1.032–10.20, P group to the very high predictive score group. We formed four increasingly serious risk categories(scores 0–3, 4–7, 8–11, and 12–14)according to the frequency associated with the different risk scores of DGF. The HMP predictive score indicates good discriminative power with a c?statistic of 0.706 in the validation cohort, and it had significantly better prediction value for DGF compared to both terminal flow(P = 0.012) and resistance(P = 0.006).Conclusion: The HMP predictive score is a good noninvasive tool for assessing the qu

关键词： Delayed Graft Function Donation after Cardiac Death Hypothermic Machine Perfusion kidney Transplantation

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Repetitive administration of cultured human CD34+cells improve adenine-induced kidney injury in mice

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World Journal of Stem Cells 2023年第4期15卷 268-280页

作者： Takayasu Ohtake Shoichi Itaba Amankeldi A Salybekov Yin Sheng Tsutomu Sato Mitsuru Yanai Makoto Imagawa Shigeo Fujii Hiroki Kumagai Masamitsu Harata Takayuki Asahara Shuzo Kobayashi Regenerative Medicine The Center for Cell Therapy&Regenerative MedicineShonan Kamakura General HospitalKamakura 247-8533KanagawaJapan kidney Disease and Transplant center Shonan Kamakura General HospitalKamakura 247-8533KanagawaJapan Regenerative Medicine Shonan Research Institute of Innovative MedicineKamakura 247-8533KanagawaJapan Kamakura Techno-science Inc. Kamakura 248-0036Japan Advanced Medicine Science Tokai University School of MedicineIsehara 259-1193Japan Department of Pathology Sapporo Tokushukai HospitalSapporo 004-0041Japan Department of Pathology Sapporo Medical CenterSapporo 004-0041Japan Human Life CORD Japan Inc Chuo-ku 103-0012TokyoJapan Cell Processing and Cell/Genome Analysis Center The Center for Cell Therapy&Regenerative MedicineShonan Kamakura General HospitalKamakura 247-8533KanagawaJapan kidney Disease and Transplant Center Shonan Kamakura General HospitalKamakura 247-8533KanazawaJapan

BACKGROUND There is no established treatment to impede the progression or restore kidney function in human chronic kidney disease(CKD).AIM To examine the efficacy of cultured human CD34+cells with enhanced proliferating potential in kidney injury in *** Human umbilical cord blood(UCB)-derived CD34+cells were incubated for one week in vasculogenic conditioning *** culture significantly increased the number of CD34+cells and their ability to form endothelial progenitor cell colony-forming *** tubulointerstitial injury of the kidney was induced in immunodeficient non-obese diabetic/severe combined immunodeficiency mice,and cultured human UCB-CD34+cells were administered at a dose of 1×106/mouse on days 7,14,and 21 after the start of adenine *** Repetitive administration of cultured UCB-CD34+cells significantly improved the time-course of kidney dysfunction in the cell therapy group compared with that in the control *** interstitial fibrosis and tubular damage were significantly reduced in the cell therapy group compared with those in the control group(Pkidney tissue was dramatically decreased in the cell therapy group compared with those in the control group(Pkidney *** administration of cultured human UCB-CD34+cells significantly improved tubulointerstitial damage in adenine-induced kidney injury in mice via vasculoprotective and anti-inflammatory effects.

关键词： Chronic kidney disease CD34+cell Adenine Tubulointerstitial injury Quality and quantity control culture Umbilical cord blood

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International kidney paired donation transplantations to increase kidney transplant of O group and highly sensitized patient: First report from India

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World Journal of Transplantation 2017年第1期7卷 64-69页

作者： Vivek B Kute Himanshu V Patel Pankaj R Shah Pranjal R Modi Veena R Shah Sayyed J Rizvi Bipin C Pal Priya S Shah Pavan S Wakhare Saiprasad G Shinde Vijay A Ghodela Umesh T Varyani Minaxi H Patel Varsha B Trivedi Hargovind L Trivedi Department of Nephrology and Clinical Transplantation Institute of Kidney Diseases and Research Center Dr HL Trivedi Institute of Transplantation Sciences (IKDRC-ITS) Department of Urology and transplantation IKDRC-ITS Department of Anesthesia IKDRC-ITS Department of Pathology Laboratory Medicine Transfusion Services and Immunohematology IKDRC-ITS

AIM To report the first international living related two way kidney paired donation(KPD) transplantation from India which occurred on 17 th February 2015 after legal per-mission from authorization committee. METHODS Donor recipient pairs were from Portugal and India who were highly sensitized and ABO incompatible with their spouse respectively. The two donor recipient pairs had negative lymphocyte cross-matching, flow cross-matchand donor specific antibody in two way kidney exchange with the intended KPD donor. Local KPD options were fully explored for Indian patient prior to embarking on international KPD. RESULTS Both pairs underwent simultaneous uneventful kidney transplant surgeries and creatinine was 1 mg/d L on tacrolimus based immunosuppression at 11 mo follow up. The uniqueness of these transplantations was that they are first international KPD transplantations in our *** International KPD will increases quality and quantity of living donor kidney transplantation. This could be an important step to solving the kidney shortage with additional benefit of reduced costs, improved quality and increased access for difficult to match incompatible pairs like O blood group patient with non-O donor and sensitized patient. To the best of our knowledge this is first international KPD transplantation from India.

关键词： kidney paired donation International kidney paired donation Living donor kidney transplantation

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Cardiac implications of thrombotic thrombocytopenic purpura

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World Journal of Cardiology 2018年第12期10卷 254-266页

作者： Szymon L Wiernek Bo Jiang Gregory M Gustafson Xuming Dai Division of Cardiology McAllister Heart Institute University of North Carolina at Chapel Hill Division of Cardiology Lang Research Center New York Presbyterian Medical Group–Queens Hospital

Thrombotic thrombocytopenic purpura(TTP) is a multisystem disorder that essentially can affect any organ in the human body. The hallmark of the pathogenesis in TTP is the large von Willebrand factor multimers on plateletmediated micro-thrombi formation, leading to microvascular *** studies showed that cardiac arrest and myocardial infarction are the most common immediate causes of death in these patients. Clinical manifestations of cardiac involvement in TTP vary dramatically, from asymptomatic elevation of cardiac biomarkers, to heart failure, MI and sudden cardiac death. There is limited knowledge about optimal cardiac evaluation and management in patients with TTP. The absence of typical cardiac symptoms,combined with complicated multi-organ involvement in TTP, may contribute to the under-utilization of cardiac evaluation and treatment. Prompt diagnosis and timely initiation of effective therapy could be critically important in selected cases. Based on our experience and this review of the literature, we developed several recommendations for focused cardiac evaluation for patients with acute TTP:(1) patients with suspected or confirmed TTP should be screened for the potential presence of cardiac involvement with detailed history and physical,electrocardiogram and cardiac enzymes;(2) clinical deterioration of TTP patients warrants immediate cardiac reevaluation;(3) TTP patients with clinical evidence of cardiac involvement should be monitored for telemetry, cardiac biomarkers and evaluated with transthoracic echocardiography. These patients require urgent targeted TTP treatment as well as cardiac-specific treatment. Aspirin therapy is indicated for all TTP patients. Since epicardial coronary artery involvement is rare, cardiac catheterization is usually not required, given the high risk for hemorrhage and kidney injury;(4) we recommend evidence-based medical therapy for ischemic symptoms and heart failure. TTP patients with evidence of cardiac invol

关键词： Thrombotic thrombocytopenic purpura Acute myocardial infarction Coronary artery disease Anti-platelet therapy von Willebrand factor ADAMTS13

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Assessment of Retreatment Tuberculosis Cases amongst All Tuberculosis Cases Notification in District Swat 2015-2017: A Retrospective Cohort Review

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Journal of Tuberculosis research 2019年第1期7卷 11-18页

作者： Zahir Shah Maqsood Ali Khan Hina Najmi Razia Fatima Aashifa Yaqoob Abdul Ghafoor Ahmed Wali Amjad Khan Shah Hasan Khan Ghulam Mustafa Halepota Provincial Tuberculosis Control Program Swat Khyber Pakhtunkhwa Pakistan Sukh Initiative Aman Health Care Services Karachi Pakistan National Tuberculosis Control Program Islamabad Pakistan Provincial Tuberculosis Control Program Quetta Pakistan Pakistan kidney and Liver Institute & research Center Lahore Pakistan

Retreatment Tuberculosis (TB) has long been a neglected area in global TB control. To determine the notification of re-treatment Tuberculosis TB cases among all enrolled TB patients, we conducted a retrospective cohort review of routine program data from 2015-2017. A total of 8663 tuberculosis cases (all new and old forms) were registered at the study site. Of these, 1916 (22%) were pulmonary bacteriological confirmed;3498 (40.37%) were pulmonary clinical diagnosed and 3396 (39.20%) were clinically or bacteriological confirmed extra-pulmonary tuberculosis cases. Retreatment cases trend was found to be increased from 1% in 2015 to 1.6% in 2017. Majority of re-treatment cases were in the age group 18 - 43 years and only 7 patients were under the age of 15 years. This concludes that notification of re-treatment cases is very low among all enrolled TB patients and this could be possibly due to the fact that most of the re-treatment cases were registered as new cases or misdiagnosed. There is need and space to increase retreatment cases. Most of the re-treatment cases were usually converted to MDR-TB (Multi-Drug Resistant Tuberculosis) as noted in PMDT (Programmatic management of Drugs resistant tuberculosis) site. Therefore, proper history and diagnosis should be encouraged at the time of case notification.

关键词： Retreatment TB Cases Retrospective Cohort Swat Pakistan

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TRP Channels:Structural and Molecular Basis of the Assembly of the TRPP2/PKD1 Complex

TRP Channels:Structural and Molecular Basis of the Assembly ...

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“基因、进化与生理功能多样性”海内外学术研讨会暨中国生理学会第七届比较生理学学术会议

作者： Maximilian H.Ulbrich Albert C.M.Ong Ehud Y.Isacoff 杨建 Department of Molecular and Cell Biology University of CaliforniaBerkeleyCA 94720USA kidney Genetics group Academic Unit of NephrologySheffield Kidney InstituteSchool of Medicine and Biomedical SciencesUniversity of SheffieldSheffieldUK Department of Molecular and Cell Biology University of CaliforniaBerkeleyCA 94720USA Material Sciences & Physics Biosciences DivisionsLawrence Berkeley National LaboratoryBerkeleyCA 94720USA Department of Biological Sciences Columbia UniversityNew YorkNY 10027USA

Autosomal dominant polycystic kidney disease（ADPKD） is one of the most common genetic diseases in *** is caused by mutations in PKD1 or ***1（also known as polycystin-1 or PC1） is a 4302-amino acid,465 kDa integral membrane protein containing 11 putative transmembrane regions. Its large extracellular N terminus contains a number of well recognized repeats and domains,some of which are known to interact with extracellular matrix *** short intracellular C terminus contains a G protein activation ***,PKD1 is generally thought to function as a cell surface receptor for extracellular ligands and matrix proteins that couples extracellular stimuli,including mechanical stimuli,to intracellular signaling. TRPP2（also known as PKD2,polycystin-2 or PC2） is a member of the transient receptor potential （TRP） channel *** is a 968 amino acid,110 kDa integral membrane protein with 6 putative transmembrane segments and a pore-forming *** forms a Ca-permea- ble nonselective cation *** mutations in PKD1 and TRPP2 lead to ADPKD is *** two proteins likely participate in the same molecular and cellular processes in some kidney cells,since mutations in them produce similar pathological manifestations. Although both proteins are widely distributed, they show high levels of expression in kidney cells,where they are present on the plasma membrane and in the Golgi and endoplasmic *** also colocalize in the primary cilia of kidney *** two proteins associate physically and form functional ***,the subunit composition of this heteromeric complex and the molecular mechanism underlying its assembly are *** a combination of biochemistry, x-ray crystallography and a single molecule optical imaging method to determine the subunit composition of proteins in the plasma membrane of live cells,we find that this complex contains 3 TRPP2 and 1 PKD1. We also identified a coiled coil domain in the C terminus of TR

关键词： TRP channels Polycystic kidney disease Subunit stoichiometry Crystal structure Cross-linking Single molecule

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Nrf2是调控解毒、抗氧化、抗炎等细胞保护机制的重要转录因子--它的活性可被保健食物及其他因素增强(英文)

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生理学报 2015年第1期67卷 1-18页

作者： Martin L Pall Stephen Levine 华盛顿州立大学美国俄勒冈州波特兰97232-3312 Allergy research group 美国加利福尼亚州阿拉米达94502

转录因子Nrf2(nuclear factor erythroid-2-related factor 2)可激活人类基因组中500多种基因的转录,这些基因大多数具有细胞保护功能。Nrf2通过解毒机制产生细胞保护作用,这些机制增强了有害异物和有毒金属的解毒和排泄。Nrf2经20多种... 详细信息

转录因子Nrf2(nuclear factor erythroid-2-related factor 2)可激活人类基因组中500多种基因的转录,这些基因大多数具有细胞保护功能。Nrf2通过解毒机制产生细胞保护作用,这些机制增强了有害异物和有毒金属的解毒和排泄。Nrf2经20多种基因的作用来增加高度协调的抗氧化活性;Nrf2也具有重要的抗炎作用;Nrf2促进线粒体的生物合成抑或提高线粒体功能;Nrf2增强细胞自噬以清除毒性蛋白的聚集体和功能异常的细胞器。有益健康的营养素和其他因素,包括酚类抗氧化剂、γ-和δ-生育酚和三烯生育酚、长链Ω-3脂肪酸DHA和EPA、类胡萝卜素(其中番茄红素可能活性最强)、十字花科蔬菜中的异硫氰酸酯、葱蒜类蔬菜中的硫化物、及萜类化合物,至少部分是通过增加Nrf2活性起作用的。其他一些有益健康并增加Nrf2活性的因素包括低水平的氧化应激[毒物兴奋效应(hormesis)]、锻炼和热量限制。现已发现,增加Nrf2活性可预防和/或治疗模型动物和/或人类许多慢性炎症性疾病,包括各种心血管疾病、肾脏疾病、肺脏疾病、中毒性肝损伤疾病、癌症(预防)、糖尿病/代谢综合征/肥胖、败血症、自身免疫性疾病、炎性肠病、HIV/AIDS及癫痫。较少证据提示增加Nrf2活性可降低其他16种疾病的风险,这些疾病中的大多数可能是NO/ONOO-环有关的疾病,而Nrf2可削弱NO/ONOO-环元素的多种作用。已知最健康的饮食(地中海和冲绳地区的传统饮食)富含促进Nrf2活性的营养素,这就像我们的祖先在旧石器时代的饮食一样。Nrf2是否同时具有延长寿命和促进健康的作用是有争议的。Nrf2活性过度的可能负面作用也被讨论。Nrf2不是一个灵丹妙药,但可能对于促进健康非常重要,特别是对于那些日常暴露于有毒化学药品的人。

关键词：亲电体和氧化剂 Keap1 ERK PI3K GSK-3β AMPK 蛋白激酶C和G 有害异物和有毒金属解毒慢性炎症性疾病氧化和硝化应激

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Protective effect of modified Huangqi Chifeng decoction(加味黄芪赤风汤)on immunoglobulin A nephropathy through toll-like receptor 4/myeloid differentiation factor 88/nuclear factor-kappa B signaling pathway

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Journal of Traditional Chinese Medicine 2024年第2期44卷 324-333页

作者： LI Liusheng ZHAO Mingming CHANG Meiying SI Yuan ZHAO Jinning YANG Bin ZHANG Yu Department of Oncology Beijing Hospital of Integrated Traditional Chinese and Western MedicineBeijing 100039China Department of Nephropathy Xiyuan HospitalChina Academy of Chinese Medical SciencesBeijing 100091China Xin-Huangpu Joint Innovation Institute of Chinese Medicine Guangzhou 510070China Department of Pathology Xiyuan Hospital China Academy of Chinese Medical SciencesBeijing 100091China Department of Experimental research Center Xiyuan HospitalChina Academy of Chinese Medical SciencesBeijing 100091China

OBJECTIVE:To examine the nephroprotective mechanism of modified Huangqi Chifeng decoction(加味黄芪赤风汤,MHCD)in immunoglobulin A nephropathy(IgAN)***:To establish the IgAN rat model,the bovine serum albumin,lipopolysaccharide,and carbon tetrachloride 4 method was *** rats were then randomly assigned to the control,model,telmisartan,and high-,medium-,and low-dose MHCD groups,and were administered the respective treatments via intragastric administration for 8 *** levels of 24-h urinary protein,serum creatinine(CRE),and blood urea nitrogen(BUN)were measured in each *** alterations were *** deposition was visualized through the use of immunofluorescence *** ultrastructure of the kidney was observed using a transmission electron *** expression levels of interleukin-6(IL-6),monocyte chemoattractant protein-1(MCP-1),and transforming growth factor-β1(TGF-β1)were examined by immunohistochemistry and quantitative polymerase chain *** of toll-like receptor 4(TLR4),myeloid differentiation factor 88(MyD88),and nuclear factor-kappa B(NF-κB)P65,were examined by immunohistochemistry,Western blotting,and quantitative polymerase chain ***:The 24-h urine protein level in each group increased significantly at week 6,and worsen from then *** this process can be reversed by treatments of telmisartan,and high-,medium-,and low-dose of MHCD,and these treatments did not affect renal ***,and high-,and medium-dose of MHCD reduced IgA *** histopathology demonstrated the protective effect of high-,medium-,and low-dose of MHCD against kidney *** expression levels of MCP-1,IL-6,and TGF-β1 in kidney tissues were downregulated by low,medium and high doses of MHCD ***,treatment of low,medium and high doses of MHCD decreased the protein and mRNA levels of TLR4,MyD88,and NF-κ***:MHCD exerted nephroprotective effects on IgAN rats,and MHCD regulated

关键词： glomerulonephritis IGA toll-like receptor 4 myeloid differentiation factor 88 NF-kappa B signal transduction inflammation renal fibrosis modified Huangqi Chifeng decoction

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