蛋白质组学在结核分枝杆菌研究中的应用

蛋白质组学是在基因组学基础上发展起来的新兴学科, 其基本技术包括样品制备、蛋白质分离和蛋白质鉴定分析, 其中的核心技术是双向凝胶电泳技术(2-Dimensional Electrophoresis, 2-DE)和质谱技术(Mass Spectrometry, MS)。近年来, 蛋白质组学技术已应用于结核分枝杆菌的研究领域。应用蛋白质组学技术分离、鉴定、检测结核分枝杆菌致病株的全菌蛋白及分泌蛋白, 分析其蛋白组成, 可深入解析结核分枝杆菌的致病机理和耐药机制。通过对结核分枝杆菌致病株抗原的分析, 为研制预防结核病的新型疫苗拓展了空间。通过对结核分枝杆菌临床分离株的蛋白组成分析还发现了一些有意义的结核病早期诊断标志物。蛋白质组学技术还应用于寻找新的药物靶标, 在研制和筛选新的抗结核药物等方面展示了一些有价值的研究成果, 为更好地开展结核病的预防、早期诊断及治疗打下了基础。     

蛋白质组学技术在植物类囊体膜蛋白研究中的应用

类囊体作为植物光合作用光反应的重要场所,在植物亚细胞蛋白质组学研究中倍受关注.介绍了植物蛋白质组学相关技术,包括双向凝胶电泳(2DE)、高效液相色谱(HPLC)、高效毛细管电泳(HPCE)、质谱(MS)和蛋白质组学数据库在植物类囊体膜蛋白研究中的应用.同时对类囊体膜蛋白质组学的研究趋势进行了探讨.     

SELDI-TOF MS技术及其在癌症早期诊断中的应用

ELDI—TOF MS技术是差异蛋白质组学研究中的一种新技术。本文介绍了这种技术的特点及分析步骤,总结了SELDI—TOF MS技术在癌症早期诊断中的应用,最后指出这种技术存在的问题,并对其前景进行了展望。     

激光捕获显微切割技术结合^18O标记定量蛋白质组技术在胃癌标志物筛查中的应用研究

建立一种更加精确地分离鉴定胃癌特异肿瘤标志物的定量蛋白质组学技术.首先采用激光捕获显微切割技术(LCM)纯化胃腺癌细胞及胃黏膜良性上皮细胞,将裂解的样本总蛋白经过1D SDS-PAGE预分离,然后采用17O/16O分别标记两种样本酶切后的多肽混合物.结合纳升级液相色谱(Nano-HPLC-MS/MS)定量地鉴定胃癌细胞和胃黏膜良性上皮细胞的差异表达蛋白.共筛选出78个差异表达蛋白,其中42个蛋白质在胃癌组织中表达上调,36个蛋白质下调.Western blot技术验证了其中几个差异蛋白(moesin,periostin,annexin A2,annexin A4)的表达,与蛋白质组学研究的结果一致.LOM技术结合18O稳定同位素标记的定量蛋白质组学技术,为研究胃癌发生机制、筛选胃癌的分子标志物提供了新的思路,亦为诸如胃癌等复杂体系蛋白质的分离鉴定提供了新的技术选择.     

定量蛋白质组学在急性高原病研究中的应用进展

急性高原病(acute mountain sickness,AMS)是人体急性暴露于高原低压低氧环境后出现多系统生理紊乱的临床综合征。定量蛋白质组学技术可以系统定量并描述机体蛋白质组成和动态变化规律,近年来在多种疾病的预防、诊断、治疗和发生机制等方面研究应用广泛。本文系统综述了定量蛋白质组学技术及其在AMS的预防、诊断、治疗和急进高原习服机制研究中的应用进展,以期为AMS的发病机制、提前干预、临床治疗和AMS的蛋白质组学研究提供参考。     

艾滋病痴呆综合征蛋白质组学的研究进展

艾滋病痴呆综合征(acquired immunodeficiency syndrome dementia complex, ADC)是由人类免疫缺陷病毒(human immunodeficiency virus,HIV)侵犯中枢神经系统,造成患者意识、行为和运动能力改变为主要临床表现的综合征。约50%的HIV感染者表现出不同形式的认知功能损伤。ADC的发病机制尚未完全阐明,诊断也无明确的病理和生化指标。目前,蛋白质组学技术以其高灵敏度和高通量的特点广泛应用于传染病研究。本文综述了近年来ADC蛋白质组学研究领域的进展,包括ADC蛋白质组学研究所用样品、蛋白质组学技术及其在ADC研究中的应用,为ADC的诊断和发病机制研究提供新思路。     

基于蛋白质组学技术的新型冠状病毒肺炎研究进展

新型冠状病毒肺炎（coronavirus disease 2019,COVID-19）是一种由严重急性呼吸综合征冠状病毒2 (severe acute respiratory syndrome coronavirus 2,SARS-CoV-2)引发的传染病。此种病毒传染性强、传播速度快,对全球人民的身体健康和生命安全造成严重威胁。蛋白质组学技术以其高通量、高灵敏度的特点,在疾病生物标志物的发现、分子机制研究及治疗靶点研究中扮演着重要角色,并被广泛应用于COVID-19的研究中。本文介绍了SARS-CoV-2的基因组结构及病毒感染过程,总结了目前常用的基于质谱的蛋白质组学研究技术,重点综述了蛋白质组学技术在COVID-19生物标志物的发现、分子机制研究和药物治疗靶标研究中的应用进展,最后展望了蛋白质组学的未来发展方向,以期能够有助于推动蛋白质组学技术在COVID-19精准诊断和治疗中的发展。     

顺铂作用卵巢癌细胞株的蛋白质组学研究

应用蛋白质组学技术观察顺铂作用于人卵巢癌SKOV3细胞株后蛋白质分子的表达差 异, 探讨蛋白质组学方法在化疗药物抗肿瘤作用机制研究方面的应用. 用顺铂(6 mg/mL)作用人卵巢癌SKOV3细胞6 h, 分别收集实验组与对照组细胞, 裂解并提取细胞内总蛋白; 以固相IPG胶条(17 cm, pH4~7)为载体, 进行第一向等电聚焦和第二向SDS-PAGE电泳, 得到实验组与对照组双向电泳凝胶图; 经考马斯亮蓝染色后, 在PDQuest软件的辅助下进行实验组与对照组蛋白质表达的比较, 选择并切取明显差异点共11个, 经肽质量指纹谱(PMF)和串级质谱(MS/MS)分析, 检测出差异点中包含的主要蛋白. 通过质谱分析显示, 顺铂处理后表达明显上调的有原肌球蛋白家族、肌动蛋白家族、热休克蛋白60(HSP60)、磷酸丙糖异构酶(TIM)家族等; 表达下调的有烯醇酶家族等. 这几类蛋白质多参与细胞内能量代谢、细胞形态维持、细胞转化凋亡等生理过程, 提示顺铂的抗肿瘤机制可能与此有关.     

DIGE技术及其在植物蛋白质组学研究中的应用

双向荧光差异凝胶电泳(DIGE)是一项新出现的荧光标记定量蛋白质组学技术,比经典的双向凝胶电泳(2-DE)具有更高的动力学范围和灵敏性。文章综述了DIGE蛋白质组学的基本原理、实验方法、在植物蛋白质组学研究中的应用和局限性,并对DIGE技术的应用做了展望。     

宏蛋白质组学技术在废水生物处理工艺研究领域中的应用

随着后基因组时代的到来,宏蛋白质组学逐渐兴起并在生命科学基础领域和临床医药领域成功运用,宏蛋白质组学技术现已成为各研究领域炙手可热的方法之一.宏蛋白质组学技术在废水生物处理研究领域中的应用刚起步,但已展示其强大功能.本文主要综述近年来国内外宏蛋白质组学在废水生物处理研究领域的研究进展,回顾及总结了宏蛋白质组学的研究策略及应用,如鉴定功能性蛋白质/酶、揭示污染物的微生物降解途径、推断废水生物处理系统的关键代谢途径、及探讨不同污泥微生物群落微生态变化等.
     

microRNAs in kidneys: biogenesis, regulation, and pathophysiological roles

MicroRNAs (miRNA) are endogenously produced, short RNAs that repress and thus regulate the expression of almost half of known protein-coding genes. miRNA-mediated gene repression is an important regulatory mechanism to modulate fundamental cellular processes such as the cell cycle, growth, proliferation, phenotype, and death, which in turn have major influences on pathophysiological outcomes. In kidneys, miRNAs are indispensable for renal development and homeostasis. Emerging evidence has further pinpointed the pathogenic roles played by miRNAs in major renal diseases, including diabetic nephropathy, acute kidney injury, renal carcinoma, polycystic kidney disease, and others. Although the field of renal miRNA research is still in its infancy and important questions remain, future investigation on miRNA regulation in kidneys has the potential to revolutionize both the diagnosis and treatment of major renal diseases.     

Ionenkanalerkrankungen der Niere und Nebenniere

Genetic kidney diseases represent a significant proportion of kidney diseases manifesting in childhood and adolescence, but are also gaining importance in slowly progressive or late-onset adult diseases. A significant portion of kidney diseases particularly in childhood are associated with end stage renal disease and/or other relevant morbidity. An early (molecular) diagnosis can be a prerequisite for a better prognostic assessment and provides opportunities in terms of optimized symptomatic therapy. Mechanistically speaking, mutations in ion channel-associated nephropathy represent—in addition to structural defects of the glomerular filter (e.g., COL4A3, LAMB2, nephrin) and disorders of signaling pathways that are relevant for the development of the urogenital tract (e.g., HNF1B, WT1)—a significant proportion of the group with respect to number and prototypes. Determination of the molecular genetics of (hypokalemic) salt-losing tubulopathies has contributed significantly to our understanding of the central role of the kidney in salt balance. The spectrum of renal ion channelopathies is shown using the example of classical salt-losing tubulopathies (Bartter syndrome and Gitelman syndrome), the transient receptor potential (TRP) channel group and the role of channel changes in aldosteronism and congenital hypertension.     

The Function of MicroRNAs in Renal Development and Pathophysiology

MicroRNAs (miRNAs) are a class of endogenous small non-coding RNAs that modulate diverse biological processes predominantly by translation inhibition or induction of mRNA degradation. They are important regulatory elements involved in renal physiology and pathology. Dysregulation of miRNAs disrupts early kidney development, renal progenitor cell differentiation and the maintenance of mature nephrons. miRNAs are also reported to participate in various renal diseases, including chronic kidney disease, acute kidney injury, allograft acute rejection and renal cell carcinoma. Differentially regulated miRNAs may represent innovative biomarkers for diagnosis and prognosis. Therefore, determining the roles of miRNAs in different types of renal diseases will help to clarify the pathogenesis and facilitate the development of novel therapies.     

The Quest for Renal Disease Proteomic Signatures: Where Should We Look?

Renal diseases are prevalent and important. However, despite significant strides in medicine, clinical nephrology still relies on nonspecific and inadequate markers such as serum creatinine and total urine protein for monitoring and diagnosis of renal disease. In case of glomerular renal diseases, biopsy is often necessary to establish the diagnosis. With new developments in proteomics technology, numerous studies have emerged, searching for better markers of kidney disease diagnosis and/or prognosis. Blood, urine, and renal biopsy tissue have been explored as potential sources of biomarkers. Some interesting individual or multiparametric biomarkers have been found; however, none have yet been validated or entered clinical practice. This review focuses on some studies of biomarkers of glomerular renal diseases, as well as addresses the question of which sample type(s) might be most promising in preliminary discovery phases of candidate proteins.     

Cystic renal disease in the domestic ferret

Cystic renal diseases in domestic ferrets are a common anecdotal finding but have received scant systematic assessment. We performed a 17-y, case-control retrospective analysis of the medical records of 97 ferrets housed at our institution between 1987 and 2004, to determine the prevalence and morphotypes of cystic renal diseases in this species. Histologic sections stained with hematoxylin and eosin, Masson trichrome, or periodic acid-Schiff were evaluated by a comparative pathologist, and statistical analysis of hematologic and serum chemistry values was correlated with morphologic diagnosis. Of the 97 available records, 43 were eliminated due to lack of accompanying tissues. Of the 54 remaining cases, 37 (69% prevalence) had documented renal cysts, and 14 of the 54 ferrets (26%) had primary polycystic disease consisting of either polycystic kidney disease affecting renal tubules or, more commonly, glomerulocystic kidney disease. Secondary polycystic lesions were identified in 11 ferrets (20%), and 12 ferrets (22%) exhibited focal or isolated tubular cysts only as an incidental necropsy finding. Ferrets with secondary renal cysts associated with other developmental anomalies, mesangial glomerulopathy, or end-stage kidney disease had hyperphosphatemia and elevated BUN in comparison with those with primary cystic disease and elevated BUN compared with those without renal lesions. Although reflecting institutional bias, these results implicate primary and secondary cystic renal diseases as highly prevalent and underreported in the domestic ferret. In addition to the clinical implications for ferrets as research subjects and pets, these findings suggest a potential value for ferrets as a model of human cystic renal diseases.     

足细胞发育异常及相关肾脏疾病研究进展

足细胞是肾小球滤过屏障的重要组成部分,其数量减少或功能障碍将导致肾小球滤过功能损伤和相关肾脏疾病的发生。足细胞为不可再生性细胞,其数量和功能在一定程度上取决于其正常发育。已发表的文献和本实验室的研究工作表明,遗传或不良宫内环境等原因所致的足细胞发育不良,可能导致成年后肾小球滤过功能障碍,并成为某些胎源性肾脏疾病发生或易感的病因之一,而表观遗传学机制可能参与介导足细胞发育过程中某些关键基因的表达异常。本文对足细胞结构功能和正常发育、足细胞发育异常的病因和机制、以及足细胞发育异常所致的肾脏疾病等几方面进行综述,以期对发育源性足细胞相关肾脏疾病的诊断与治疗提供借鉴与参考。     

运动性急性肾损伤与新型肾功能生物标志物

大强度运动中,非创伤性急性肾损伤（acute kindey injury, AKI）经常发生,表现为血尿、蛋白尿、血红蛋白尿等。一般认为,中低程度的运动性急性肾损伤是可逆的,可完全恢复。但动物实验与人类研究均发现,严重的运动性肾损伤会导致“功能性”急性肾损伤发展为“结构性”急性肾损伤,并增加慢性肾病的风险。运动性急性肾损伤对机体的潜在健康威胁已引起国内外相关领域学者的广泛关注。血清肌酐 (serum creatinine, Scr)和尿量作为肾功能的传统经典标志物,不能特异性反映早期肾损伤,而新型肾损伤标志物可进一步明确损伤的位置及严重程度。在运动领域,利用新型生物标志物进行无创性检查,识别早期运动性急性肾损伤非常必要。本文综述了反映肾小球或肾小管损伤、细胞周期停滞和肾损伤修复的新型生物标志物,着重论述了尿中性粒细胞明胶酶相关脂质运载蛋白（NGAL)和肾损伤分子-1（KIM-1)与肾功能的关系,以及长时间耐力运动、急性运动和高强度间歇阻力运动3种运动形式对肾功能的影响,旨在引起重视,精准识别风险,及时进行早干预。     

运动性急性肾损伤与新型肾功能生物标志物

大强度运动中,非创伤性急性肾损伤（acute kindey injury, AKI）经常发生,表现为血尿、蛋白尿、血红蛋白尿等。一般认为,中低程度的运动性急性肾损伤是可逆的,可完全恢复。但动物实验与人类研究均发现,严重的运动性肾损伤会导致“功能性”急性肾损伤发展为“结构性”急性肾损伤,并增加慢性肾病的风险。运动性急性肾损伤对机体的潜在健康威胁已引起国内外相关领域学者的广泛关注。血清肌酐 (serum creatinine, Scr)和尿量作为肾功能的传统经典标志物,不能特异性反映早期肾损伤,而新型肾损伤标志物可进一步明确损伤的位置及严重程度。在运动领域,利用新型生物标志物进行无创性检查,识别早期运动性急性肾损伤非常必要。本文综述了反映肾小球或肾小管损伤、细胞周期停滞和肾损伤修复的新型生物标志物,着重论述了尿中性粒细胞明胶酶相关脂质运载蛋白（NGAL)和肾损伤分子-1（KIM-1)与肾功能的关系,以及长时间耐力运动、急性运动和高强度间歇阻力运动3种运动形式对肾功能的影响,旨在引起重视,精准识别风险,及时进行早干预。     

Role of kidney biomarkers of chronic kidney disease: An update

Chronic kidney disease (CKD) is a progressive pathological condition marked by deteriorating renal function over time. Diagnostic of kidney disease depend on serum creatinine level and glomerular filtration rate which is detectable when kidney function become half. The detection of kidney damage in an early stage needs robust biomarkers. Biomarkers allow monitoring the disease progression at initial stages of disease. On the onset of impairment in cellular organization there is perturbation in signaling molecules which are either up-regulated or down-regulated and act as an indicator or biomarker of diseased stage. This review compiled the cell signaling of different kidney biomarkers associated with the onset of chronic kidney diseases. Delay in diagnosis of CKD will cause deterioration of nephron function which leads to End stage renal disease and at that point patients require dialysis or kidney transplant. Detailed information on the complex network in signaling pathway leading to a coordinated pattern of gene expression and regulation in CKD will undoubtedly provide important clues to develop novel prognostic and therapeutic strategies for CKD.