Proteomic profiling and protein identification by 2-DE-MS in the liver of Carassius auratus exposed to Yongcheng coalmine subsidence area

Proteomic profile of the Carassius auratus liver was constructed to evaluate contamination degree of aquatic organism exposed to coalmine subsidence area. The results revealed that 55 proteins were up-regulated and 51 down-regulated among 160 differentially expressed proteins. These proteins were mainly involved in the physiological activities, such as granzyme B signaling, dermatan sulfate biosynthesis and cytotoxic T lymphocyte mediated apoptosis of target cells. Meanwhile, they were related to two network interaction modules. One was cell adhesion and migration, angiogenesis, DNA repair, and the other was small molecule metabolism, structural molecule activity and defense response. Ingenuity Pathway Analysis (IPA) was further utilized to predict differential proteins molecular activation, including tnf, tp53, Anxa3, fgf2 and mvp. The result sheds light on liver protein expression profiles of C. auratus, and provides important data for controlling and managing water pollution from heavy metals.     

颈动脉粥样硬化、血管内皮舒张功能与冠心病的相关性分析

目的研究分析冠心病患者颈动脉粥样硬化与血管舒张功能的相关性。方法选取2012年12月～2013年12月期间收治的45例冠心病患者为试验组,并选取同期体检的45例无冠心病患者为对照组。应用高频超声检查全部患者的血管内皮舒张功能（EDD）、颈动脉斑块数、颈动脉内一中膜厚度（IMT）以及颈动脉斑块积分。结果试验组患者的颈动脉斑块数、IMT、颈动脉斑块积分均大于对照组,差异有统计学意义（P〈0．05）;试验组患者EDD明显小于对照组,差异有统计学意义（P〈0．05）。结论血管内皮舒张功能、颈动脉粥样硬化与冠心病的发病具有紧密联系,两者是临床预防冠心病的关键指标。     

血清hs-CPR浓度和冠脉病变复杂程度的相关性

目的探讨冠心病患者血清超敏C反应蛋白(hs-CRP)水平与冠脉病变复杂程度的关系。方法对140例冠脉造影患者,按照冠脉造影下冠脉病变复杂程度将所有患者分为单支病变组(n=36)、双支病变组(n=41)、多支病变组(n=28)及对照组(n=35)采用免疫透射比浊法测患者血清的hs-CPR水平,根据冠脉造影结果行Gensini评分。结果血清hs-CRP和Gensini积分在不同冠脉病变组间差异有显著性,且随着病变血管支数增加而增高(P0.01)。血脂、hsCRP与Gensini积分呈显著正相关(r=0.314,P0.001),hsCRP是Gensini积分的独立影响因子。结论血清hs-CRP浓度与冠脉病变程度及危险因素有相关性,对冠心病患者的病变预测有较好的临床意义。     

Impairment of endothelium-dependent relaxation in aorta from rats with arteriosclerosis induced by excess vitamin D and a high-cholesterol diet.

The present investigation was undertaken to characterize the relaxing responsiveness in aortic strips from rats with arteriosclerosis, which was produced by excess vitamin D2 (VD) administration followed by treatment with or without a high-cholesterol diet for 6 weeks (VD + CHOL and VD group, respectively). This arteriosclerotic aorta was characterized by medial calcification and intimal cell proliferation. Helical strips of thoracic aorta were suspended in oxygenated Krebs-Henseleit solution. Under precontraction with noradrenaline, endothelium-dependent relaxations to acetylcholine were significantly attenuated in the VD and VD + CHOL as compared with the control. Relaxation to calcium ionophore A23187 was also significantly attenuated in the VD + CHOL. However, the relaxing responses to acetylcholine and A23187 in aortas from rats fed a high-cholesterol diet alone remained unaffected. Nitroglycerin caused an equal degree of relaxation in aortas from control and arteriosclerotic rats. There was a significant negative correlation between the relaxing response to acetylcholine and the calcium content in the aorta. These results indicate that in arteriosclerotic rat aortas, the endothelium-dependent relaxation to acetylcholine is impaired in proportion to the degree of calcification, and such impairment is facilitated by cholesterol feeding but can not be attributed to hypercholesterolemia or vascular cholesterol deposition.     

高通量分析紫杉醇在动物体内的药动学及组织分布

目的 建立基于96孔板的高通量测定SD大鼠血浆和荷瘤小鼠组织中紫杉醇浓度的方法.方法 将SD大鼠血浆样品、荷瘤小鼠组织样品及内标物多西他赛置于96孔板中,采用96/384道半自动液体移液器取液进行高通量的液液萃取,高效液相色谱-串联质谱（LC/MS/MS）法测定各生物样品的紫杉醇浓度.结果 紫杉醇的线性范围为2 ～ 500 μg/L,最低定量下限为2μg/L.在SD大鼠血浆中,紫杉醇的半衰期、平均滞留时间、药时曲线下面积、药时曲线下总面积、药物清除率、表观分布容积分别为（6.3 ±0.4）h、（7.16 ±0.13）h、（3 070 ±201）（h· μg）/L、（3246±199）（h· μg）/L、（1.55±0.09）L/（h·kg）、（13±8）L/kg.荷瘤小鼠尾静脉注射紫杉醇注射液7.5 mg/kg后0.5、1、2、4、8h,紫杉醇在小鼠心、肝、脾、肺、肾、肿瘤中含量差异有统计学意义（P＜0.05）.结论 LC/MS/MS法专一、准确、稳定、简便并且实现了高通量分析,适用于紫杉醇的SD大鼠药动学和荷瘤小鼠组织分布研究.     

单次大剂量水杨酸钠注射所致大鼠耳蜗基因表达谱改变

目的:研究大剂量水杨酸钠耳毒性的作用机制。方法:运用寡核苷酸基因芯片技术分析大剂量水杨酸钠注射对大鼠耳蜗基因表达谱的影响。联用TRIzol和RNA纯化柱分别从13周正常大鼠和水杨酸钠注射后的大鼠耳蜗内容物提取总RNA,将总RNA逆转录为cDNA,再将cDNA体外转录为cRNA,在此过程中同时进行生物素标记,将cRNA片段化后作为探针与质量检测芯片杂交,确定无问题后再与寡核苷酸基因芯片杂交扫描,用Affymetrix Microarray Suite software 5.0分析处理扫描结果,获得两种情况下耳蜗的基因表达谱。结果:表达上调2倍以上的基因和EST有42个,下调2倍以上的有49个。结论:水杨酸钠影响各种离子通道和突触蛋白基因,可能有助于解释水杨酸钠所致的耳聋和耳鸣作用机制,而其他变化基因亦有助于解释水杨酸钠广泛的药理作用。     

Microminipig, a non-rodent experimental animal optimized for life science research:novel atherosclerosis model induced by high fat and cholesterol diet

Atherosclerotic lesions were observed in male and ovariectomized female Microminipig (MMP) fed a high fat and cholesterol diet with sodium cholate (HFCD/SC) for 3 months. HFCD/SC induced hypercholesterolemia accompanied by an increase in serum total cholesterol (T-Cho), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), and cholesterol ester (CE). Unlike the mouse or rabbit, a dominant LDL-C fraction in the intact MMP, similar to that in humans, was observed by serum lipoprotein analysis. HFCD/SC increased body weight gain. At the end of the experiment, computed tomography scans of conscious animals showed that HFCD/SC had decreased liver attenuation values (Hounsfield unit) and increased subcutaneous and abdominal fat, suggesting the induction of fatty liver and obesity. HFCD/SC induced atherosclerotic lesions in systemic arteries, including the external and internal iliac arteries, abdominal aorta, coronary artery, and cerebral arterial circle. Atherosclerosis and pathological findings induced by HFCD/SC in MMP were similar to those in humans. The MMP is a potentially suitable tool for investigating human atherosclerosis.     

Attainment of optional low-density lipoprotein cholesterol goal of less than 70 mg/dl and impact on prognosis of very high risk stable coronary patients: a 3-year follow-up

Objectives: We aimed to investigate the proportion of very high-risk patients with coronary heart disease (CHD) who achieve the optional low-density lipoprotein cholesterol (LDL-C) target of < 70 mg/dl (1.8 mmol/liter), the factors that influence the success rate and the impact on their prognosis.

Research design and methods: We enrolled 1337 consecutive patients with stable CHD. Fasting lipids were determined and all cardiovascular events were recorded during a median follow-up of 33 months.

Results: The majority (86.5%) of patients were taking lipid-lowering medication (95.5% statins), but only 50.6% had LDL-C levels of < 100 mg/dl (2.6 mmol/liter). In total, 941 (70.4%) patients were considered very high risk and only 15.1% of them had LDL-C levels of < 70 mg/dl. Τhe use of intensive lipid-lowering medication was associated with 12-fold (95% CI 6.98 – 20.76; p < 0.001) higher possibility in achieving LDL-C levels of < 70 mg/dl. Attainment of LDL-C levels of < 70 mg/dl by patients at very high risk were independent predictors of all cardiovascular events (HR = 0.34, 95% CI 0.17 – 0.70; p = 0.003).

Conclusions: The vast majority of very high-risk patients do not achieve the optional LDL-C goal; this is mainly due to the suboptimal uptitration of statin dose and is translated into loss of clinical benefits.     

二烯丙基二硫诱导人胃癌MGC803细胞分化的差异蛋白质分析

目的研究二烯丙基二硫(diallyl disulfide,DADS)诱导人胃癌MGC803细胞分化过程中蛋白质的差异表达。方法用固相pH梯度双向凝胶电泳分离DADS处理前后人胃癌MGC803细胞总蛋白,凝胶经过银染后用PDQuest软件进行分析,差异蛋白质点经胶内原位酶解,用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)测定肽质量指纹图(PMF),将所得的数据进行生物信息学处理。结果 DADS处理MGC803细胞前后的总蛋白质的双向电泳银染图谱,经扫描成像及PDQuest软件分析,对照组与处理组蛋白质点与平均匹配率分别为(576±14)个和(583±4)个与76%和70%,421个匹配,200个未被匹配。经相关数据库查询,在初步鉴定的差异蛋白质点中,发现24个与细胞分化、肿瘤转移、细胞凋亡、细胞周期、细胞免疫及代谢等相关的蛋白质,如gastric mucin、nM23、CDC2、uPAR、LIMK、RORα、MHC DR-beta-1 chain、TCR等,这些蛋白质可能在胃癌的发生发展中起着潜在的作用。结论 DADS可能通过多种途径诱导MGC803细胞分化。蛋白质组差异表达的初步分析为进一步研究胃癌分化的相关蛋白质及标记物奠定一定基础。     

胆维丁原料中共晶与游离态维生素D3和胆固醇含量的定量分析

通过差示扫描量热分析(DSC)、拉曼光谱及粉末X射线衍射(PXRD)发现胆维丁原料中存在共晶现象,并对其进行表征.采用粉末X-射线衍射(PXRD)建立了共晶与游离态维生素D3和胆固醇含量的定量分析方法.     

牛磺酸对维生素D_3和尼古丁诱导的大鼠钙化血管精氨酸/NO途径的影响

目的　观察钙化血管精氨酸 /NO途径的改变和外源性牛磺酸 (taurine,Tau)对血管钙化及精氨酸 /NO途径的影响。方法　维生素D3 (vitaminD3 ,VitD3 )肌注和尼古丁(nicotine ,Nic)灌胃制备大鼠血管钙化模型 ,检测血管钙含量和血管碱性磷酸酶 (ALP)活性、血浆精氨酸和亚硝酸盐(NO-2 )含量、血管环磷酸鸟苷 (cGMP)含量、血管一氧化氮合酶 (NOS)活性及血管精氨酸 (arginine ,Arg)转运。结果　钙化组 (VDN)大鼠血管钙含量较对照组升高 6 6倍 (P <0 0 1) ,血管ALP活性高 12 6倍 (P <0 0 1) ;血浆NO生成减少 ,血管cGMP水平降低 ,血管NOS活性增高 ,其中cNOS活性高 18 9% (P <0 0 1) ,但精氨酸转运在血管平滑肌和内皮明显减少 (- 6 3 8% ,- 5 5 2 % ,P <0 0 1)。口服牛磺酸治疗的大鼠较单纯VDN组 ,血管钙含量降低 4 9 5 % (P <0 0 1) ,血管ALP活性明显降低 ,血浆NO生成增加 ,血管cGMP含量增加 2 9 1% (P均 <0 0 1) ,血管精氨酸转运在血管平滑肌和内皮分别增加 79 4 %和 5 5 1% (P <0 0 1)。结论　给予外源性牛磺酸可以减轻VitD3 加Nic大鼠的血管钙化程度 ,改善钙化血管L Arg/NOS/NO/cGMP途径紊乱 ;提示牛磺酸对防治动脉粥样硬化等疾病的血管钙化可能具有潜在临床意义     

实时三维超声及二维斑点追踪技术对拟诊冠心病患者局部室壁运动的评价

【摘要】 目的 探讨实时三维超声（RT-3DE）及二维斑点追踪成像（2D-STI）技术检测拟诊冠心病患者局部室壁运动功能的临床应用价值。 方法 选取临床拟诊为冠心病、二维超声示室壁运动正常的住院患者 143 例, 根据冠脉造影左冠状动脉主干（LM）和（或）左前降支（LAD）和（或）左回旋支（LCX）病变结果分为狭窄≥75%组（A 组）73 例和狭窄＜ 75%组（B 组）70 例。 应用 RT-3DE 及 2D-STI 分别检测并比较 2 组患者左室节段射血分数（sEF）和收缩期纵向峰值应变（SL）, 利用 ROC 曲线比较 2 种技术诊断冠心病的曲线下面积。 结果 （1）A 组病变冠脉所支配室壁的 sEF 及 SL均较 B 组减小（P＜ 0.05）;（2）节段平均 sEF 与节段平均 SL呈正相关（r=0.689, P＜ 0.05）;（3）两项指标均显示异常的左冠供血各节段 sEF 对诊断供血冠脉狭窄≥75%的曲线下面积大于 S（ L 最大曲线下面积分别为 0.922 和 0.874）。 结论 RT-3DE 及 2D-STI 均可评估冠心病患者的心肌局部运动状况, 进而推测冠脉病变程度, 但 RT-3DE 指标可能更优于 2D-STI。     

五参口服液对糖尿病合并冠心病大鼠Apelin及心肌组织病理的影响

目的:观察五参口服液对糖尿病合并冠心病大鼠血浆及心肌组织的Apelin含量及心肌组织病理学的影响。方法:按三步骤叠加法复制糖尿病合并冠心病模型。取DM模型大鼠,随机分为模型组、硝酸异山梨醇酯片组、格列美脲组、五参口服液大剂量组、小剂量组,分别灌胃给药,每日1次,连续8周,同时高脂饲料喂养,末次给药后,禁食12 h,以40 mg.kg-1戊巴比妥钠液腹腔注射麻醉,固定测量心电图,再经腹主动脉采血,分离血浆,处死动物,取心室心肌组织,以ELISA法检测血浆和心肌组织的Apelin含量;取心肌组织,HE染色,镜检。结果:与正常组比较,模型组大鼠血浆及心肌组织Apelin含量显著降低(P<0.01),心肌组织出现病理学损害;与模型对照组比较,五参口服液大剂量组、消心痛组大鼠血浆及心肌组织Apelin含量显著升高(P<0.05,P<0.01),心肌组织病理学损害明显减轻。结论:五参口服液对糖尿病合并冠心病缺血性心肌损害具有一定的保护作用。     

环磷酸腺苷葡胺联合丹参对老年冠心病合并充血性心力衰竭患者心功能及B型利钠肽的影响

目的观察环磷腺苷葡胺联合丹参治疗老年冠心病合并充血性心力衰竭的疗效及对B型利钠肽(B-typy natriuretic peptide,BNP)的影响,并探讨其作用机制。方法 60例老年冠心病合并充血性心力衰竭心功能Ⅲ～Ⅳ级住院患者,随机分为治疗组和对照组,每组30例。对照组在充血性心力衰竭治疗中采用加强休息、限盐、利尿、扩血管、给予血管紧张素转化酶抑制剂等常规治疗,但不使用环磷腺苷葡胺及丹参治疗。治疗组除常规治疗外,环磷腺苷葡胺60～160mg加入5%葡萄糖注射液250ml中,丹参注射液20ml+5%葡萄糖液250ml静脉滴注,1次/d,静脉滴注,14d为1疗程。给药前及治疗后分别测定血浆BNP浓度,并观察美国纽约心脏协会心功能改善状况。结果治疗14d后,治疗组总有效率90.00%,对照组总有效率73.33%,2组差异有统计学意义(P<0.05)。2组治疗后BNP和LVEF与治疗前比较,差异有统计学意义(P<0.05或<0.01);2组间比较,差异有统计学意义(P<0.05或<0.01)。结论环磷腺苷葡胺联合丹参是治疗老年冠心病合并充血性心力衰竭有效的药物。     

Allergenicity study of EGFP-transgenic chicken meat by serological and 2D-DIGE analysis

Genetically modified (GM) foods must be tested for safety, including by allergenicity tests to ensure that they do not contain new allergens or higher concentrations of known allergens than the same non-GM foods.     

Upregulation of 14-3-3 isoforms in acute rat myocardial injuries induced by burn and lipopolysaccharide

1. Burn-induced myocardial injuries can be acute due to loss of body fluid and blood redistribution, and subacute due to pathogenic toxins of infecting bacteria. The goal of this study was to examine expression of 14-3-3 in the injured myocardium. 2. Myocardial injury models were created in vivo by subjecting rats to severe burn and administration of lipopolysaccharide. RT-PCR and Western blotting were employed to assess the expression of 14-3-3 proteins and messenger ribonucleic acid (mRNA) for 14-3-3eta and gamma in the myocardium, respectively. 3. In the two models, we found that 14-3-3 proteins were induced in a time-dependent fashion. Such a change is at least in part attributed to increases in mRNAs for 14-3-3gamma and eta. In contrast to 14-3-3xi, whose mRNA was not detectable in the heart, mRNA for 14-3-3gamma was found significantly elevated between 24-48 h after burn. 14-3-3eta mRNA exhibited a marked increase at 3 h continuing to 12 h and then decreased nearly to a normal level after 48 h. In lipopolysaccharide-treated intact rats, 14-3-3gamma mRNA in myocardium showed a significant increase, reaching a peak at 4 h, followed by a decrease at 6 h. In contrast, 14-3-3eta mRNA had a slight increase without significance. 4. Our results suggest that 14-3-3 may play a role in both acute and subacute (postburn infectious) phases of severe burn.     

一种新的高盐致高血压动物模型及其血管重构改变

目的 :研究单纯高盐饲养对正常Wistar大鼠血压及血管结构的影响 ,并建立一种新的高盐致高血压大鼠模型。方法 :雄性Wistar大鼠 ,于哺乳期 (3周龄 )后 ,随机分成 2组 :正常盐 (含 0 .5 %NaCl饲料 )饲养组和高盐 (含 4 %NaCl饲料 )饲养组 ,持续饲养 18周 ,定期测量鼠尾收缩压和体重。实验结束时 ,经右颈动脉插管测平均动脉压。在血管灌注固定后 ,胸主动脉、肾动脉及肠系膜小动脉作形态学测量。结果 :高盐与正常盐饲养组间 ,大鼠体重在分组前后均无区别。高盐饲料饲养后第 8周起 ,高盐组鼠尾血压明显高于正常盐饲养组 ,并持续至实验结束 ,且随时间增加 ,血压增高更明显。颈动脉平均压测量结果进一步证实高盐饲养组血压明显高于正常盐饲养组。检测大、中、小血管的血管中层厚度、中层厚度与管腔比率、血管壁横截面积 ,高盐饲养组均明显较正常盐饲养组增大 ,但两组间血管腔径无区别。高盐饲养组左心室重量指数明显高于正常盐饲养组。结论 :长期高盐饲养可致正常Wistar大鼠血压升高。故单纯高盐饮食在正常生理状态下的Wistar大鼠可形成一种新的高血压模型 ,且高血压形成同时伴随有循环系统心脏和血管的重构现象。该高血压模型可能较盐敏感性高血压模型更接近人类的高盐摄入与高血压病的实际情况。