安徽地区猪囊尾蚴蛋白质理化学及免疫学分析

应用ＳＤＳ－聚丙烯酰胺凝胺电泳、化学染色和免疫印迹技术对安徽地区猪囊尾蚴蛋白质进行了理化学和免疫学分析。猪囊尾蚴蛋白质至少有２８条区带，其中有１２条主带，在１２条主带中有７条能被囊虫病人血清识别，其分子量分别为９２、６６、５４、５１、３４．５、３０和１２ｋＤ，其中除３４．５ｋＤ为脂蛋白，其余均为糖蛋白。３０ｋＤ囊尾蚴抗原分子与肝吸虫病和日本血吸虫病血清有交叉反应，其他６个主要血清学抗原与上述吸虫病     

Purification, morphology and partial characterization of a reovirus-like agent associated with neonatal calf diarrhea

Studies have been conducted on a virus which has recently been implicated as an etiological agent in widely disseminated cases of calf diarrhea. The virus was isolated from feces and intestinal mucosa from experimentally infected calves and from cell cultures which had been inoculated with material from infected calves. The virus was purified by differential centrifugation, nuclease treatment, extraction with Genetron 113 and cesium chloride (CsC1) gradient. Purified and non-purified samples were studied by electron microscopy. Both types of preparations contained unenveloped particles approximately 64 nm in diameter with a hexagonal core region 36 nm in diameter. Subunits of the capsids appeared to be arranged in accordance with cubic synmetry. Essential lipids were not associated with the virus. The buoyant density of purified virus was 1.359. Viral nucleic acid was determined by chemical methods to be ribonucleic acid (RNA). The morphology and some chemical characteristics appear to be similar to those of the reovirus group. However, the exact classification of this virus and its relationship to other groups remains to be established.     

Purification and partial characterization of a new group of gastric cancer associated antigens.

The corresponding antigens of MG series monoclonal antibodies (MG5, MG9, MGd1 and MGe1) against gastric cancer were purified and partially characterized. Each of these monoclonal antibodies was purified by passing through a DEAE-52 cellulose columns and covalently coupled with CNBr-activated sepharose 4B successively. By means of concanavalin A and antibody affinity chromatography, the corresponding antigens of MG series McAb were extracted from gastric cancer tissue respectively. Immunological and biochemical studies confirmed that the corresponding antigens of MG series McAb were a new group of gastric cancer associated neutral glycolipid and glycoprotein antigens.

     

钙周期素结合蛋白在乳腺癌转移性腋淋巴结中表达的预后价值

目的研究乳腺癌转移性腋淋巴结中钙周期素结合蛋白(Cacy BP/SIP)的表达与患者复发转移及预后的关系。方法采用免疫组化SP法检测Cacy BP/SIP在乳腺癌术后腋淋巴结中的表达,分析其表达与患者复发转移及预后的相关性。结果 Cacy BP/SIP在转移性腋淋巴结中高表达(34.88%),腋淋巴结无转移者无表达。在转移性腋淋巴结中复发转移组Cacy BP/SIP的阳性表达率46.67%,无复发转移组的阳性表达率7.70%。Cacy BP/SIP在转移性腋淋巴结中的表达与患者的生存率呈负相关性,与累积复发转移风险呈正相关性。结论 Cacy BP/SIP可能参与并促进乳腺癌的复发转移、影响患者生存。     

Progerin作用的伴侣蛋白和核纤层蛋白问的相互作用:在早老症中Mel18与emerin的相互作用

Objective :The Hutchinson-Gilford progeria syndrome (HGPS or progeria) is a childhood disorder with features of premature aging and is caused by mutations in the lamin A gene resulting in the production of an abnormal protein, termed progerin. To investigate the underlying pathogenic mecha-nism, we studied the nuclear co-localization and association of progerin interactive partner proteins (PIPPs) with lamina proteins. Methods:Both wild-type (WT) and progeria fibroblasts were studied by various methods including eonfocal microscopy, immunopreeipitation and Western blot. Results:All PIPPs discovered so-far co-localized with lamin A/C. In addition, the PIPPs were selectively associated with lamina proteins. An increased immunofluorescent staining signal was found for Mel18 in HGPS as com-pared to WT cells. An association of Me118 with emerin was observed in HGPS, but not in WT cells.Conclusion: Based on these findings, we propose that PIPPs, along with associated lamina proteins may form a pathogenic progerin-containing protein complex.     

粘细菌转录因子FruA结合蛋白敲除分析

目的：构建粘细菌发育特异性转录因子FruA相关蛋白FruB的敲除菌株，初步阐明FruB在粘细菌发育周期中的作用。方法：在fruB基因中央编码区插入四环素抗性基因。导入野生型粘细菌DZF1后通过双交叉同源重组途径阻断基因组内fruB的功能。结果：抗性筛选和PCR检测显示分离出了fruB基因阻断菌株fruB∷TcΩ2,fruB功能缺失延缓子实体的发育及营养细胞向粘孢子分化。结论：fruB与粘细菌发育分化存在相关性。     

Sa抗原相关蛋白质的研究

目的 研究类风湿关节炎（RA）特异性抗Sa抗体识别的Sa抗原,探讨Sa抗原的相关蛋白质。方法 从人胎盘组织阴离子交换柱层析法纯化Sa抗原,应用免疫印迹法检测155例各种风湿病患者血清的抗Sa抗体,155例患者中类风湿关节炎71例,强直性脊柱炎11例、银屑病关节炎7例、反应性关节炎7例、幼年特发性关节炎4例、骨性关节炎5例、风湿性多肌痛6例、痛风6例、系统性红斑狼疮7例、干燥综合征10例、炎性肌病8例、混合性结缔组织病4例、白塞病6例、成人Still病3例。切取与免疫印迹法阳性条带相对应的电泳胶内条带,胰蛋白酶胶内酶解,用高效液相色谱-电喷雾-离子阱质谱鉴定目标蛋白质。以大肠杆菌重组表达目标蛋白质作为抗原,以免疫印迹法检测相应抗体。瓜氨酸化修饰重组蛋白质作为抗原,以免疫印迹法检测相应抗体。结果 抗Sa抗体表现为相对分子质量为50000（和）55000的阳性印迹条带,在155例风湿病中,对RA的诊断敏感性和特异性分别为47．9％和95．2％。质谱鉴定Sa抗原相关蛋白质为波形蛋白（Vimentin）。抗波形蛋白抗体在RA组与其他风湿病组的阳性率差异有统计学意义（P=0．005）,对RA的诊断敏感性和特异性分别为36．6％和83．3％,但与抗Sa抗体的一致性差（Kappa=0．316）。抗瓜氨酸化波形蛋白抗体阳性率在RA组较其他风湿病组高（P〈0．01）,对RA的诊断敏感性和特异性分别为49．3％和86．9％,且与抗Sa抗体的一致性较高（Kappa=0．746）。结论 运用质谱技术鉴定的Sa抗原相关蛋白质波形蛋白,瓜氨酸化修饰后的抗原性与Sa抗原相仿,提示瓜氨酸化波形蛋白与Sa抗原密切相关。     

Evagination of Taenia solium cysticerci: a histologic and electron microscopy study

BACKGROUND: After a human being ingests a cysticercus, the larval stage of Taenia solium, the cysticercus gradually develops toward the adult parasite. In this paper, we describe the sequential progress of evagination of cysticerci. METHODS: Intact cysticerci were obtained from swine muscle, and incubated in bovine bile to stimulate evagination. Dissecting, light, and electron microscopy of whole parasites and histologic sections were used for photographic registers. RESULTS: The first event was the widening of the opening of the bladder wall for the scolex and neck to emerge. The two chambers that conform the cysticercus were identified. Histologic sections provided explanation for the conformation of the spiral canal.The scolex uncoils during evagination but does not turn inside out. CONCLUSIONS: The scolex and the neck comprise a different structure from the bladder wall, although they are contiguous.     

Effects of a 5-lipoxygenase-activating protein inhibitor on biomarkers associated with risk of myocardial infarction: a randomized trial

Context  Myocardial infarction (MI) is the leading cause of death in the world. Variants in the 5-lipoxygenase–activating protein (FLAP) gene are associated with risk of MI. Objective  To determine the effect of an inhibitor of FLAP on levels of biomarkers associated with MI risk. Design, Setting, and Patients  A randomized, prospective, placebo-controlled, crossover trial of an inhibitor of FLAP (DG-031) in MI patients who carry at-risk variants in the FLAP gene or in the leukotriene A₄ hydrolase gene. Of 268 patients screened, 191 were carriers of at-risk variants in FLAP (87%) or leukotriene A₄ hydrolase (13%). Individuals were enrolled in April 2004 and were followed up by designated cardiologists from a university hospital in Iceland until September 2004. Interventions  Patients were first randomized to receive 250 mg/d of DG-031, 500 mg/d of DG-031, 750 mg/d of DG-031, or placebo. After a 2-week washout period, patients received DG-031 if they had received placebo first or placebo if they had received DG-031 first. Treatment periods lasted for 4 weeks. Main Outcome Measures  Changes in levels of biomarkers associated with risk of MI. Results  In response to 750 mg/d of DG-031, production of leukotriene B₄ was significantly reduced by 26% (95% confidence interval [CI], 10%-39%; P = .003) and myeloperoxidase was significantly reduced by 12% (95% CI, 2%-21%; P = .02). The higher 2 doses of DG-031 produced a nonsignificant reduction in C-reactive protein (16%; 95% CI, –2% to 31%; P = .07) at 2 weeks. However, there was a more pronounced reduction (25%; 95% CI, 5%-40%; P = .02) in C-reactive protein at the end of the washout period that persisted for another 4 weeks thereafter. The FLAP inhibitor DG-031 was well tolerated and was not associated with any serious adverse events. Conclusion  In patients with specific at-risk variants of 2 genes in the leukotriene pathway, DG-031 led to significant and dose-dependent suppression of biomarkers that are associated with increased risk of MI events.      

人表皮光老化相关蛋白的蛋白质组分析与鉴定

目的:以老年人正常皮肤为研究对象,分析曝光部位及非曝光部位的表皮差异蛋白,筛选出表皮细胞中与皮肤光化相关的蛋白.方法:从分离所得的不同部位老年人表皮中提取蛋白质,并进行2-DE分离,PDQUEST软件分析凝胶上差异蛋白点,进一步采用MALDI-TOF质谱分析及数据库查询,鉴定差异表达蛋白;并在蛋白质水平进行Western-blotting验证差异候选蛋白.结果:通过分析比较两组表皮双向电泳图谱,确定了15个差异蛋白点,经MALDI-TOF分析及数据库查询,成功鉴定了其中5个差异表达蛋白,发现热休克蛋白B1(Heat shock protein B1,HSPB1)、膜联蛋白A2(Annexin A2,ANXA2)、角蛋白80(Keratin,typeⅡ cytoskeletal 80,K2C80)、丝裂原激活的蛋白激酶3(Mitogen- activated protein kinase 3,MK03)在老年人曝光部位的表皮中高表达,而谷胱甘肽S转移酶A2(Glutathione S-transferase A2,GSTA2)在非曝光部位表皮中高表达.结论:老年人正常皮肤曝光部位和非曝光部位的表皮存在差异蛋白,经分析和鉴定的5种蛋白质与皮肤的光老化有密切的关系,为进一步研究表皮细胞在光老化过程中的作用及分子机制奠定基础.     

Genetic characterization and protein stability analysis of a Chinese family with Von Hippel-Lindau disease

Background Von HippeI-Lindau disease （VHL）,a heritable autosomal dominant disease characterized by neoplasia in multiple organ systems,has rarely been reported in Asia.We genetically investigated a unique Chinese family with VHL disease and performed an analysis of the VHL protein stability.Methods Genomic deoxyribonucleic acid （DNA） extracted from peripheral blood was amplified by polymerase chain reaction （PCR） to three exons of the VHL gene in 9 members of the Chinese family with VHL disease.PCR products were directly sequenced.We estimated the effects of VHL gene mutation on the stability of pVHL,which is indicated by the free energy difference between the wild-type and the mutant protein （△△G）.Results The Chinese family was classified as VHL type 1.Three family members,including two patients and a carrier,had a T to G heterozygotic missense mutation at nucleotide 515 of the VHL gene exon 1.This missense mutation resulted in the transition from leucine to arginine in amino acid 101 of the VHL protein.There was low stability of the VHL protein （the △△G was 12.71 kcal/mol） caused by this missense mutation.Conclusions We first reported a family with this VHL gene mutation in Asia.This missense mutation is predicted to significantly reduce the stability of the VHL protein and contribute to the development of the renal cell carcinoma （RCC） phenotype displayed by this family.The genetic characterization and protein stability analysis of families with VHL disease are important for early diagnosis and prevention of the disease being passed on to their offspring.     

喉部分切除术治疗喉癌30年回顾

目的：探讨30年间喉部分切除术在喉癌治疗中的应用。方法：30年间行喉部分切除术257例,将30年喉部分切除术后治疗喉癌分前10年、中间10年和后10年3个阶段,进行对比观察。前10年、中间10年和后10年喉癌手术中喉部分切除术分别占9．6％、52．3％和68．6％。结果：30年间喉部分切除术257例,前10年、后10年和后10年3年生存率分别为50．0％、63．0％和73．6％,5年生存率分别为37．5％、55．6％和71．8％;术后1－3月内拔管率分别为37．5％、50．6％和71．4％,术后咽瘘发生率分别为25．0％、12．3％和3．0％。结论：30年间喉部分切除术患者年龄有逐渐增高趋势;全喉切除术逐渐减少,喉部分切除术逐渐增加;3、5年生存率和术后拔管率有逐渐提高的趋势;术后咽瘘和逐渐减少的趋势。     

与人巨细胞病毒UL132编码蛋白相互作用蛋白的初步筛选和鉴定

目的：探讨应用酵母双杂交系统筛选出与人巨细胞病毒（HCMV）UL132编码蛋白相互作用的人胎脑cDNA文库组织蛋白,阐明其在先天性巨细胞病毒感染中可能的致病机制。方法：采用聚合酶链式反应扩增HCMV UL132片段,酶切、纯化扩增的目的片段及酵母诱饵表达载体pGBKT7,连接HCMV UL132片段到载体pGBKT7上,将连接后的重组体pGBKT7-UL132转化到酵母菌AH109中,再将人胎脑文库DNA也转化到酵母AH109中,筛选与HCMVUL132编码蛋白相互作用的人胎脑文库蛋白,并对阳性克隆进行测序和生物信息学分析。结果：成功构建诱饵表达载体pGBKT7-UL132,双酶切鉴定可见800及7500bp2条目的条带;转化文库质粒后计算转化效率为6.6×10³ cfu·μg^-1,显色反应显示有95个菌落呈蓝色,最终确认10个克隆与HCMV UL132编码蛋白相互作用,Blast结果显示7个克隆与CAML高度同源。结论：利用酵母双杂交系统成功筛选出与HCMVUL132编码蛋白相互作用的人胎脑cDNA文库蛋白CAML,推测该目的蛋白可能在HCMV感染所致神经系统损伤、病毒的入侵和增殖过程中发挥重要作用。     

S100钙结合蛋白P在胃癌患者胃癌组织和血清中的表达及其临床意义

目的: 研究胃癌患者胃癌组织和血清中S100钙结合蛋白P(S100P)表达及其与临床病理参数的关系,监测原发肿瘤切除前后血清S100P蛋白水平,探讨其与预后的关系。 方法: 选取手术切除并经病理检查确诊的胃癌患者50例作为研究对象,所有患者均可获取术后石蜡组织标本及术前血清,其中20例患者可获取新鲜胃癌组织及配对癌旁组织,另30例患者可获取术后动态血清。半定量RT-PCR法检测组织中S100PmRNA水平;免疫组织化学法及ELISA法分别检测组织和血清中S100P蛋白水平。选择同期30名体检者为正常对照组,ELISA法检测血清S100P蛋白水平。结果: 胃癌组织S100P mRNA表达水平明显低于癌旁组织(P<0.05);胃癌组织S100P蛋白阳性表达率低于癌旁组织(P<0.05),S100P表达下调与脉管癌栓有关联(P<0.05)。胃癌患者术前血清S100P水平低于正常对照组,但差异无统计学意义(P>0.05);无淋巴结转移胃癌患者血清S100P水平高于有淋巴结转移患者,但差异无统计学意义(P> 0.05);胃癌患者手术后2周血清S100P水平明显低于手术前(P<0.05)。结论: S100P在胃癌患者胃癌组织和血清中表达水平下调,其表达水平与预后有关联。持续动态监测血清S100P水平变化可能预测复发及判断预后。     

ERCP治疗胰腺部分切除术后胰瘘1例

<正>1临床资料患者,女,71岁。入院2个月前,因上腹部闷痛不适行腹部超声检查提示胰腺占位性病变。为求行胰腺肿物手术治疗于2018年4月8日来大连医科大学附属第二医院。近期体重未见明显变化。既往体健。肝肾功能良好;肿瘤标志物：CA199:3.61U/mL;IG4：阴性;心肺功能检查良好。术前核磁共振检查提示：胰腺颈体交界部可见大小约1.6cm×1.8cm占位性病变,T1表现为低信号;T2表现为稍高信号;核磁增强动脉期呈乏血供改变,静脉期有环周强化（图1）;核磁共振诊断：胰腺占位性病变,自身免疫性胰腺炎与胰腺癌待鉴别。     

糖尿病大鼠心肌损伤中程序性坏死通路相关蛋白的表达

目的 探讨程序性坏死（Necroptosis）通路相关蛋白在糖尿病心肌中的表达及其可能机制。 方法 在大鼠高糖高脂饮食喂养基础上腹腔注射链脲佐菌素40 mg/kg制备糖尿病模型大鼠,其中正常组（CON组,n=6）、高脂高糖组（HC组,n=6）和糖尿病组（DM组,n=6）;10周后超声检测左心室舒张末期内径（LVEDD）、左心室收缩末期内径（LVESD）及左心室射血分数（LVEF）;处死大鼠后测量心质量（HW）、体质量（BW）、心体比（HW/BW）;ELISA法检测血液中肿瘤坏死因子-α（TNF-α）、白细胞介素6（IL-6）及丙二醇（MDA）水平;心尖部组织行Masson染色;RT-PCR及Western blotting检测心肌组织中RIPK1、RIPK3、MLKL的含量变化。 结果 与CON组比,HC组HW、TNF-α、IL-6、MDA水平增加,RIPK1、RIPK3和MLKL的蛋白表达量增加（P < 0.05~P < 0.01）,但MLKL的mRNA表达量无明显变化（P>0.05）;与HC组比较,DM组HW/BW升高,LVEF下降,TNF-α、IL-6及MDA水平增加,RIPK1、RIPK3、MLKL的mRNA及蛋白含量增加（P < 0.01）,Masson染色蓝染加重。 结论 高糖状态可诱导心肌损伤,增加心肌纤维化及氧化水平,可能与上调Necroptosis通路有关。     

Metrics associated with NIH funding: a high-level view

Objective

To introduce the availability of grant-to-article linkage data associated with National Institutes of Health (NIH) grants and to perform a high-level analysis of the publication outputs and impacts associated with those grants.

Design

Articles were linked to the grants they acknowledge using the grant acknowledgment strings in PubMed using a parsing and matching process as embodied in the NIH Scientific Publication Information Retrieval & Evaluation System system. Additional data from PubMed and citation counts from Scopus were added to the linkage data. The data comprise 2 572 576 records from 1980 to 2009.

Results

The data show that synergies between NIH institutes are increasing over time; 29% of current articles acknowledge grants from multiple institutes. The median time lag to publication for a new grant is 3 years. Each grant contributes to approximately 1.7 articles per year, averaged over all grant types. Articles acknowledging US Public Health Service (PHS, which includes NIH) funding are cited twice as much as US-authored articles acknowledging no funding source. Articles acknowledging both PHS funding and a non-US government funding source receive on average 40% more citations that those acknowledging PHS funding sources alone.

Conclusion

The US PHS is effective at funding research with a higher-than-average impact. The data are amenable to further and much more detailed analysis.