Proteomic analysis of rat pheochromocytoma PC12 cells

PC12 cell line is well documented and widely applied as many kinds of models in neurobiological and neurochemical studies. Yet a thorough proteomic analysis has not been performed so far. Here we report the construction of a large-scale 2-D protein database for PC12 cells. The proteins extracted from PC12 cells were separated by 2-DE and identified by MALDI-TOF/TOF MS. A total of 1080 protein spots, excised from three different 2-D gels, were identified with high confidence. These proteins represent 474 different gene products, mainly binding proteins and enzymes. Three hundred and seven identified protein spots were located in the low-molecular weight region below 20 kDa. This database today represents one of the largest 2-D databases for higher eukaryotic cell proteomes and for low-molecular weight proteins. In addition, fragment ion spectra obtained by TOF/TOF confirmed that calcylin in PC12 cells was N-acetylated. The database of PC12 proteome is expected to be a powerful tool for neuroscientists.     

Two‐dimensional proteome reference map of Prototheca zopfii revealed reduced metabolism and enhanced signal transduction as adaptation to an infectious life style

Biochemical, serological, and genetic analyses have identified two genotypes of Prototheca zopfii, a unicellular microalga belonging to the family Chlorellaceae. The P. zopfii genotype 1, abundantly present in cow barns and environment, remains nonpathogenic, while P. zopfii genotype 2 has been isolated from cows with bovine mastitis. The present study was carried out to identify the protein expression level difference between the pathogenic and nonpathogenic strains of P. zopfii. A total of 782 protein spots were observed on the 2D fluorescence difference gel electrophoresis (2D DIGE) gels among which 63 and 44 proteins were identified to be overexpressed in genotypes 1 and 2, respectively. The limited number of protein entries specific for Prototheca in public repositories resulted mainly in the identification of proteins described in other algae, microorganisms, or plants. Gene ontology (GO) analysis indicated reduced carbohydrate metabolism in genotype 1, while genotype 2 displayed enhanced DNA binding, kinase activity, and signal transduction. These effects point to metabolic and signaling adaptations in the pathogenic strain and provide insights into the evolution of otherwise highly similar strains. All MS data have been deposited in the ProteomeXchange with identifier PXD000126.     

微生态制剂(贝飞达)治疗非便秘型肠易激综合征

目的探讨微生态制剂贝飞达对肠易激综合征（IBS）的疗效。方法采用中国预防医学科学院流行病学研究所海斯药业有限公司生产的双歧三联活菌胶囊贝飞达（长双歧杆菌、嗜酸乳酸杆菌、粪肠球菌）治疗25例IBS患者,与服用安慰剂组25例进行疗效对比分析。结果IBS特征性症状评分治疗前后的差值,治疗组明显高于安慰剂组,各种症状的改善率明显优于对照组。结论贝飞达的短期疗法有望成为IBS治疗中的一个颇有前途的手段。     

酪酸梭菌治疗肠易激综合征疗效的Meta分析

目的 系统评价酪酸梭菌治疗肠易激综合征（irritable bowel syndrome,IBS）的临床疗效。方法 计算机检索PubMed、Embase、the Cochrane Library、维普、万方、中国知网和中国生物医学文献数据库,收集建库至2019年5月1日前国内外公开发表的关于酪酸梭菌治疗IBS的随机对照试验。由两位研究员独立进行筛选、资料提取及质量评价,采用RevMan 5.3软件进行Meta分析。结果 最终纳入19篇随机对照试验,共2 194例患者。Meta分析结果显示：酪酸梭菌辅助治疗IBS时,联合治疗组的总有效率明显高于对照组（93.4% vs 77.1%,OR=4.32,95% CI 3.09‒6.04,P<0.05）;而单用酪酸梭菌治疗IBS疗效并不优于常规药物（78.8% vs 68.9%,OR=2.05,95% CI 0.96‒4.38,P=0.06）;且联合用药短期复发率显著降低（14.6% vs 29.2%,OR=0.41,95% CI 0.20‒0.85,P=0.02）。纳入的所有研究均无严重不良反应发生。结论 单用酪酸梭菌与常规药物治疗IBS相比,酪酸梭菌并未显示出更好的疗效。而采用口服酪酸梭菌辅助治疗IBS可显著提高患者的临床疗效、降低复发率,且安全性好。     

粪菌移植治疗难治性肠易激综合征的效果观察

观察粪菌移植(FMT)对难治性肠易激综合征的治疗效果, 为该病的治疗提供参考。

招募我院32例难治性肠易激综合征患者并分为治疗组(n=17)和对照组(n=15), 两组患者均给予马来酸曲美布汀片口服, 治疗组进一步通过鼻肠管行FMT。分别于治疗1个月和3个月时评估两组患者临床疗效及安全性, 并严格记录不良反应。

治疗组患者FMT治疗前、治疗1个月和治疗3个月时IBS-SSS评分和粪便分型逐渐降低(均P < 0.05), 而对照组患者这3个时间点IBS-SSS评分和粪便分型差异无统计学意义(均P > 0.05)。FMT治疗前两组患者IBS-SSS评分和粪便分型差异无统计学意义(均P > 0.05)。治疗1个月和治疗3个月时, 两组患者IBS-SSS评分差异均有统计学意义(均P < 0.05)。治疗组患者FMT治疗前、治疗1个月和治疗3个月时IBS-QOL和HAD评分逐渐降低(均P < 0.05), 而对照组患者这3个时间点的IBS-QOL和HAD评分差异无统计学意义(均P > 0.05)。FMT治疗前, 两组患者IBS-QOL和HAD评分差异无统计学意义(均P > 0.05)。治疗1个月时两组患者IBS-QOL评分差异有统计学意义(P < 0.05), 而HAD评分差异无统计学意义(P > 0.05)。治疗3个月时两组患者IBS-QOL和HAD评分差异均有统计学意义(均P < 0.05)。治疗组患者症状缓解率高于对照组(88.2% vs 53.3%)。治疗3个月后两组患者均未出现明显不良反应且未发现传染性疾病。

FMT对难治性肠易激综合征有一定的临床疗效, 且安全可靠。

     

复方嗜酸乳杆菌片对肠易激综合征患者的疗效

目的 探讨复方嗜酸乳杆菌片对肠易激综合征（IBS）患者的治疗效果。方法 根据罗马Ⅲ诊断标准选择患者并进行分组,其中便秘型IBS（C-IBS）治疗组28例,C-IBS对照组28例;腹泻型IBS（D-IBS）治疗组57例,D-IBS对照组57例。C-IBS治疗组患者应用复方嗜酸乳杆菌片联合莫沙必利进行治疗,C-IBS对照组单纯应用莫沙必利治疗。D-IBS治疗组应用复方嗜酸乳杆菌片联合应用匹维溴铵进行治疗,D-IBS对照组单纯应用匹维溴铵治疗。治疗4周后观察各组患者疗效。结果 C-IBS治疗组有效率高于C-IBS对照组（P＜0.05）;D-IBS治疗组有效率高于D-IBS对照组（P＜0.05）。结论 IBS患者应用复方嗜酸乳杆菌片联合胃肠动力药的治疗效果优于单用胃肠动力药。     

Peppermint oil in irritable bowel syndrome

In a literature search 16 clinical trials investigating 180-200 mg enteric-coated peppermint oil (PO) in irritable bowel syndrome (IBS) or recurrent abdominal pain in children (1 study) with 651 patients enrolled were identified. Nine out of 16 studies were randomized double blind cross over trials with (n = 5) or without (n = 4) run in and/or wash out periods, five had a randomized double blind parallel group design and two were open labeled studies. Placebo served in 12 and anticholinergics in three studies as comparator. Eight out of 12 placebo controlled studies show statistically significant effects in favor of PO. Average response rates in terms of "overall success" are 58% (range 39-79%) for PO and 29% (range 10-52%) for placebo. The three studies versus smooth muscle relaxants did not show differences between treatments hinting for equivalence of treatments. Adverse events reported were generally mild and transient, but very specific. PO caused the typical GI effects like heartburn and anal/perianal burning or discomfort sensations, whereas the anticholinergics caused dry mouth and blurred vision. Anticholinergics and 5HT3/4-ant/agonists do not offer superior improvement rates, placebo responses cover the range as in PO trials. Taking into account the currently available drug treatments for IBS PO (1-2 capsules t.i.d. over 24 weeks) may be the drug of first choice in IBS patients with non-serious constipation or diarrhea to alleviate general symptoms and to improve quality of life.     

肠道菌群失衡诱发肠易激综合征的机制

肠易激综合征（irritable bowel syndrome,IBS）是一种常见的功能性胃肠道疾病,严重地威胁着人类的健康与生存质量。最近的研究发现IBS的发病机制是复杂多样的,尽管其确切的发病原因尚不完全清楚,但有证据显示IBS可能与肠道菌群失衡有关。本文就有关肠道菌群的功能、IBS患者肠道菌群的特点、肠道菌群失衡导致IBS发病的可能机制的研究进展作一综述,旨在为IBS的早期诊断与有效治疗提供有价值的理论依据。     

凝结芽孢杆菌活菌片对腹泻型肠易激综合征患者的治疗

目的 观察和评价凝结芽孢杆菌活菌片（商品名：爽舒宝）对腹泻型肠易激综合征患者的临床疗效。方法 将100例腹泻型肠易激综合征患者随机分为3组,其中爽舒宝治疗组32例,马来酸曲美布汀治疗组35例,联合治疗组33例。3组患者疗程均为60 d,记录3组患者腹痛、腹泻、排便异常等症状的变化。结果 在近期疗效方面,联合治疗组有效率（87.87%）高于爽舒宝治疗组（75.00%）,而这两组的有效率均高于马来酸曲美布汀治疗组（51.43%）。在远期疗效方面,爽舒宝治疗组患者有效率（100.00%）和联合治疗组（100.00%）之间无显著差异,而这两组的有效率均高于马来酸曲美布汀治疗组（74.29%）。治疗结束后,3组患者大便次数、腹痛、大便性状等各项症状体征积分均较治疗前显著降低（P0.05）,但均低于马来酸曲美布汀治疗组（P<0.05）。结论 凝结芽孢杆菌活菌片治疗腹泻型肠易激综合征疗效显著,值得在临床治疗中推广应用。     

Proteomic analysis of cold adaptation in a Siberian permafrost bacterium--Exiguobacterium sibiricum 255-15 by two-dimensional liquid separation coupled with mass spectrometry

Bacterial cold adaptation in Exiguobacterium sibiricum 255-15 was studied on a proteomic scale using a 2-D liquid phase separation coupled with MS technology. Whole-cell lysates of E. sibiricum 255-15 grown at 4 degrees C and 25 degrees C were first fractionated according to pI by chromatofocusing (CF), and further separated based on hydrophobicity by nonporous silica RP HPLC (NPS-RP-HPLC) which was on-line coupled with an ESI-TOF MS for intact protein M(r) measurement and quantitative interlysate comparison. Mass maps were created to visualize the differences in protein expression between different growth temperatures. The differentially expressed proteins were then identified by PMF using a MALDI-TOF MS and peptide sequencing by MS/MS with a MALDI quadrupole IT TOF mass spectrometer (MALDI-QIT-TOF MS). A total of over 500 proteins were detected in this study, of which 256 were identified. Among these proteins 39 were cold acclimation proteins (Caps) that were preferentially or uniquely expressed at 4 degrees C and three were homologous cold shock proteins (Csps). The homologous Csps were found to be similarly expressed at 4 degrees C and 25 degrees C, where these three homologous Csps represent about 10% of the total soluble proteins at both 4 degrees C and 25 degrees C.     

Proteomic profiling of Cronobacter turicensis 3032, a food‐borne opportunistic pathogen

Members of the genus Cronobacter are opportunistic pathogens for neonates and are often associated with contaminated milk powder formulas. At present little is known about the virulence mechanisms or the natural reservoir of these organisms. The proteome of Cronobacter turicensis 3032, which has recently caused two deaths, was mapped aiming at a better understanding of physiology and putative pathogenic traits of this clinical isolate. Our analyses of extracellular, surface‐associated and whole‐cell proteins by two complementary proteomics approaches, 1D‐SDS‐PAGE combined with LC‐ESI‐MS/MS and 2D‐LC‐MALDI‐TOF/TOF MS, lead to the identification of 832 proteins corresponding to a remarkable 19% of the theoretically expressed protein complement of C. turicensis. The majority of the identified proteins are involved in central metabolic pathways, translation, protein folding and stability. Several putative virulence factors, whose expressions were confirmed by phenotypic assays, could be identified: a macrophage infectivity potentiator involved in C. turicensis persistence in host cells, a superoxide dismutase protecting the pathogen against reactive oxygen species and an enterobactin‐receptor protein for the uptake of siderophore‐bound iron. Most interestingly, a chitinase and a metalloprotease that might act against insects and fungi but no casein hydrolysing enzymes were found, suggesting that there is an environmental natural habitat of C. turicensis 3032.     

布拉酵母菌辅助治疗腹泻型肠易激综合征疗效的Meta分析

目的 系统评价布拉酵母菌辅助治疗腹泻型肠易激综合征(IBS-D)的有效性.方法 计算机检索The Cochrane Library、PubMed、EMbase、中国生物医学文献数据库、中国知网、维普中文期刊数据库和万方数据库,时限为建库至2020年2月.由2位研究人员按纳入、排除标准筛选文献,提取数据,评价纳人文献的方...     

微生态制剂干预肠易激综合征患者疗效meta分析

目的对微生态制剂干预肠易激综合征患者的疗效进行系统性分析,指导临床用药。方法联机检索Medline、维普、万方数据库、CNKI、CBMdisc中微生态制剂治疗肠易激综合征疗效的随机对照临床试验,对文献质量进行严格评价和资料提取,对符合质量标准的RCT进行Meta分析。结果共有7项研究符合入选标准,其中5项研究提示微生态制剂干预肠易激综合征患者疗效不优于安慰剂,2项研究认为微生态制剂有效改善患者的临床症状,荟萃结果表明微生态制剂可以有效缓解肠易激综合征临床症状[OR(95%CI):1.66(1.27,2.18)]。结论微生态制剂可以有效缓解肠易激综合征临床症状。     

携带IL-10基因的双歧杆菌对 感染后肠易激综合征的治疗作用

目的探讨转基因双歧杆菌对感染后肠易激综合征(PI-IBS)小鼠的治疗作用。方法 50只成年雄性SD小鼠,随机选取10只作为空白对照组(NC组),其余40只PI-IBS模型小鼠随机分为IBS组(IBS组)、双歧杆菌治疗组(BL组)、空质粒双歧杆菌治疗组(BL-0组)和BL-hIL-10治疗组(每组10只)。NC组和IBS组每天予0.2mL PBS灌胃1次;其余三组每天灌胃溶有经0.2%L-Arb诱导24h后的BL、BL-0、BL-hIL-10菌液(双歧杆菌浓度达5×108个/mL)的PBS液0.2mL,每天1次。所有小鼠干预7d后处死,然后行血液及结肠组织学检验。结果 (1)通过评估小鼠疾病活动指数、小鼠脾脏指数和结肠组织HE染色病理学变化,证明口服BL-hIL-10能显著减轻PI-IBS小鼠的炎症症状;(2)BL-hIL-10能升高IL-10比例并降低IL-12的比例,维持Th平衡。结论口服转基因双歧杆菌可以缓解PI-IBS小鼠的炎症反应,其机制可能是通过调节Th平衡来实现。     

Proteomic analysis of the excretory-secretory proteins of the Trichinella spiralis L1 larva, a nematode parasite of skeletal muscle

Trichinella spiralis is an intracellular nematode parasite of mammalian skeletal muscle. Infection of the muscle cell leads to the formation of a host-parasite complex that results in profound alterations to the host cell and a re-alignment of muscle-specific gene expression. The role of parasite excretory-secretory (ES) proteins in mediating these effects is currently unknown, largely due to the difficulty in identifying and assigning function to individual proteins. In this study, a global proteomics approach was used to analyse the ES proteins from T. spiralis muscle larvae. Following 2-DE of ES proteins,MALDI-TOF-MS and LC-MS/MS were used to identify the peptide spots. Specific Trichinella EST databases were assembled and used to analyse the data. Despite the current absence of a Trichinella genome-sequencing project, 43 out of 52 protein spots analysed were identified and included the major secreted glycoproteins. Other novel proteins were identified from matches with sequences in the T. spiralis database. Our results demonstrate the value of proteomics as a tool for the identification of Trichinella ES proteins and in the study of the molecular mechanism underpinning the formation of the host-parasite complex during Trichinella infections.     

经肠镜导入益生菌对肠易激综合征治疗的可行性研究

目的 观察经肠镜导入益生菌对肠易激综合征（IBS）患者症状的影响及可能的不良反应。方法 选择福建医科大学附属第一医院就诊的IBS患者,入选患者均符合IBS罗马IV诊断标准并同意进入本研究。患者肠镜检查完成时于右半结肠单次喷洒33.6 mg/mL的双歧杆菌三联活菌‒生理盐水混悬液100 mL,第0、2和4周进行粪便Bristol分型、腹痛视觉评分（VAS）、结肠症状评分和IBS严重程度调查表评分（IBS-SSS）;第0、4周进行医院焦虑抑郁量表（HADS）测定。结果 入选患者中共有5例IBS便秘型（IBS-C）、13例IBS腹泻型（IBS-D）患者完成研究,年龄分别为（49.4±10.0）岁和（36.1±8.9）岁,男女比例分别为1∶0.66和3.3∶1,均无严重不良反应发生。IBS-C组患者症状指标均无显著改善（P>0.05）,该组患者提前终止。IBS-D组患者综合疗效指数0.692,与第0周相比,第2和4周患者的粪便Bristol分型、腹痛VAS评分、症状总积分和IBS-SSS评分均显著下降（P0.05）。结论 经肠镜单次双歧杆菌三联活菌混悬液喷洒可安全有效地改善IBS-D患者的肠道症状,但对IBS-C患者无效。     

Proteomic analysis of berry-sizing effect of GA3 on seedless Vitis vinifera L

Gibberellin (GA) is widely used in the table grape and raisin industries to enlarge the berries of seedless varieties. However, the mechanism underlying its berry‐sizing effect is poorly understood. In this study, clusters of Centennial Seedless (Vitis vinifera L.) were treated with 30 ppm GA₃ on day 12 after flowering, and berries were sampled at development stages I, II and III for proteomic analysis. Among the 1479 proteins detected on 2‐DE maps, 19, 70 and 69 spots in stages I, II and III, respectively, showed an at least twofold difference in volume between treatments and controls. Of these, 125 proteins were successfully identified and assigned to eight functional groups, chief among them are metabolism and energy, stress response, expression regulation and cytoskeleton proteins. Stress‐response proteins were predominantly down‐regulated in GA₃‐treated berries in stages I and II, and significantly up‐regulated in stage III. Up‐regulation of cytoskeleton, cell‐wall modification and other important proteins was found in the two latter stages of berry development. Our proteomic results and subsequent validation revealed, for the first time, the role of redox homeostasis in GA₃‐induced berry enlargement and markedly remodeled cellular protein expression in treated berries.     

肠道菌群与肠易激综合征相关研究进展

肠易激综合征(irritable bowel syndrome,IBS)是一种功能性肠病,目前已有研究表明IBS的致病机制、症状的产生和持续时间可能与肠道菌群失调有关,且不同IBS亚型患者体内的肠道菌群种类有差异。IBS的发病机制尚不完全清楚,临床表现存在较大的个体差异。IBS的诊断和治疗尚未形成统一的标准。但已经有较多研究提示IBS可能与胃肠道动力、内脏高敏感性、肠道免疫反应和模式识别受体等密切相关,并且可以通过益生菌、益生元、抗生素、粪菌移植和饮食习惯等调节肠道菌群的失调,从而改善IBS的症状。这也为未来治疗IBS提供了新的思路。本文就肠道菌群的概况、IBS患者肠道菌群的特点、肠道菌群失衡导致IBS发病的可能机制以及IBS肠道菌群的相关治疗方法的研究进展作一综述。     

肠易激综合征模型WHBE兔神经-体液-内分泌系统调节的变化

目的观察肠易激综合征（IBS）模型WHBE兔神经内分泌激素和电解质水平的变化,探讨神经-体液-内分泌系统在IBS中的作用。方法以湿热环境应激加小剂量致泻剂诱导WHBE兔建立IBS模型,在造模9d和14d时取血分离血清,检测血清中5-羟色胺（5-HT）、多巴胺（DA）、促肾上腺皮质激素（ACTH）、皮质醇（COR）、β-内啡肽（β-EP）激素和K^＋、Na^＋、cl^-含量的变化,并以JW兔作对照。结果与正常对照组比,造模9d时WHBE兔的5-HT、DA、ACTH、β-EP、COR均显著升高（P〈0．05,P〈0．01）,Na^＋、K^＋含量均显著降低（P〈0．01）,而JW兔ACTH、β-EP、COR均显著升高（P〈0．05,P〈0．01）,K^＋含量显著降低（P〈0．05）;造模14d时WHBE兔5-HT、ACTH、β-EP均显著升高（P〈0．05,P〈0．01）,JW兔ACTH显著升高（P〈0．05）;相关分析显示,IBS模型中血清中5-HT、COR与Na^＋、K^＋浓度呈显著负相关（P〈0．05,P〈0．01）,血清中DA、β-EP水平与Na^＋、K^＋、Cl^-浓度呈显著负相关（P〈0．05,P〈0．01）,ACTH水平则与K^＋浓度呈显著负相关（P〈0．01）。结论IBS的发生不仅与HPA轴兴奋性升高有关,亦与体液因素有关,故神经-体液-内分泌系统参与了WHBE兔IBS的发生过程。