Expression of cathepsin C and tumor necrosis factor-α in human coronary artery and their relationships with coronary heart disease

AIM To explore the possible mechanism of cathepsin C （CTSC） and tumor necrosis factor-α （TNF-α） in coronary heart disease （CHD） by detecting the protein expression of CTSC and TNF-α in human coronary artery tissue. METHODS The coronary artery tissues from 52 cases of CHD and 25 cases of accidental death without any heart disease in the Forensic Judicial Expertise Center of Guizhou Medical University from October 2018 to December 2019 were collected as CHD group and control group, respectively. The coronary artery stenosis and intimal plaque formation were examined by histopathology, the protein expression of CTSC and TNF-α was determined by Western blot, and the intracellular expression of CTSC and TNF-α was analyzed by immunohistochemical staining. RESULTS The results of HE staining showed that the intima of coronary artery in control group was smooth, and no thickening or stenosis was observed. In CHD group, the intima thickened irregularly, atherosclerotic plaques formed, the intima became thinner and the lumen showed eccentric stenosis in varying degrees （P<0.05）. The results of Western blot showed that the expression of CTSC and TNF-α in CHD group was significantly higher than that in control group （P<0.05）. Immunohistochemical staining showed that both CTSC and TNF-α were expressed in the cytoplasm of foam cells, and their positive expression was significantly higher than that in control group （P<0.05）. The results of Pearson moment correlation analysis showed that there was positive correlation between the expression of CTSC and TNF-α in CHD （r²=0.743, P<0.05）. CONCLUSION The up-regulated expression of CTSC in coronary artery tissue may promote the expression of TNF-α and affect the occurrence and development of CHD.     

黄芪多糖在LPS诱导的DF-1细胞炎症反应中的抗炎作用及其调节机制

为探讨黄芪多糖(Astragalus polysaccharide,APS)在LPS(lipopolysaccharide,LPS)诱导的鸡胚成纤维细胞系DF-1(chicken embryonic fibroblast cell line,DF-1)炎症模型中对IL-1β和TNF-α表达的影响,以及细胞因子信号转导抑...     

玉米转录因子基因ZmPTF1在转基因烟草中的表达

为了探讨转录因子基因ZmPTF1在玉米种子萌发中的作用,本研究根据GenBank中收录的玉米PTF1序列设计PCR引物,利用普通玉米自交系A318克隆得到ZmPTF1的全长cDNA序列.该序列开放阅读框包括1 446bp碱基,由481个氨基酸组成.本文构建植物表达载体PCAMBIA3301-ZmPTF1转化烟草W38,经过抗生素筛选和PCR鉴定得到转基因阳性烟草,对2株生长正常的T1转基因烟草进行southern杂交分析及RT-PCR检测,结果表明外源ZmPTF1已经整合进了烟草基因组中并且能正常转录并遗传.与对照相比,转基因烟草种子的发芽率、发芽势及萌发种子α-淀粉酶活性显着增高,表明ZmPTF1基因过表达会促进烟草种子提前解除休眠进入发芽阶段,提高烟草种子发芽势和发芽率.该研究结果为进一步研究ZmPTF1基因的功能提供理论依据.     

美国豆芋花总皂苷制备及其α-葡萄糖苷酶抑制活性

为高效利用美国豆芋花皂苷类化合物,以皂苷得率和纯度为指标,研究美国豆芋花总皂苷的提取工艺及大孔树脂纯化工艺,并对制备的美国豆芋花总皂苷的抗氧化及α-葡萄糖苷酶活性抑制能力进行研究。结果表明,美国豆芋花总皂苷的提取工艺为提取溶剂90%乙醇,料液比1∶20,温度70℃,提取时间2 h。得到的提取物石油醚脱脂后用水饱和正丁醇萃取,D-101大孔树脂吸附柱纯化的工艺为上样皂苷质量浓度1 mg·m L~(-1),上样量40 m L·g~(-1)树脂,洗脱剂为80%乙醇(pH值6),洗脱剂用量4 BV(树脂柱体积)。制备的美国豆芋花总皂苷样得率4.24%,纯度达65.35%,比粗提物(纯度15.27%)提高了约3倍;抗氧化性能(清除DPPH和ABTS自由基IC50值分别为51.08、29.24μg·m L~(-1))和α-葡萄糖苷酶活性抑制能力(IC50值为83.62μg·m L~(-1))也均优于粗提物(清除DPPH、ABTS自由基IC50值分别为286.51、69.92μg·m L~(-1),α-葡萄糖苷酶的抑制浓度(IC50)值为1 043.57μg·m L~(-1)),表明该工艺可用于高活性美国豆芋花总皂苷的富集和纯化。此外,美国豆芋花总皂苷抑制α-葡萄糖苷酶活性的抑制类型为竞争型抑制,抑制常数为49.68μg·m L~(-1)。本研究结果为开发具有降血糖功能的医药中间体或功能性食品提供了新资源和实践基础。     

甘草总黄酮对金黄色葡萄球菌的作用研究

为了研究甘草总黄酮对金葡菌的抗菌活性以及亚抑菌浓度的甘草总黄酮对金黄色葡萄球菌α-溶血素分泌的影响。首先,采用微量肉汤稀释法测定了甘草总黄酮对6株金葡菌标准菌株的最小抑菌浓度;其次,采用溶血试验、蛋白免疫印迹试验以及荧光定量PCR试验分别在表型功能、蛋白表达和基因转录3个层次确证了其对α-溶血素表达的影响。结果表明：甘草总黄酮具有良好的抗金葡菌作用,其最小抑菌浓度范围为4~16 μg/mL,而亚抑菌浓度的甘草总黄酮能浓度依赖性地降低金葡菌α-溶血素的分泌。     

优质甘蓝型油菜黔油17号亲本原种的保优繁殖技术研究

通过优质甘蓝型油菜隐性核不育系98-116 AB与父本ZW 99004单株成对杂交,并在F1代杂交组合进行配合力测定,获得高配合力的成对株系材料。根据优质杂交油菜新品种黔油17号母本98-116 AB及父本ZW 004的自身特点,分别在人工隔离和自然隔离条件下进行其原原种及原种的保优繁殖技术研究,保证亲本原始的优良种性,确保大面积生产中杂交种的质量以及品种的生产潜力。     

酸性α-醋酸萘酯酶染色法的改良

通过对酸性α-醋酸萘酯酶染色过程中的几个重要因素进行研究,确定了其最佳处理条件,并探讨了几个因素间的相互作用。结果表明,固定液用2.5%戊二醛的效果要优于甲醛-丙酮混合液;用pH值5.8～6.2的孵育液孵育1.5h,甲基绿复染1～10s效果最佳。变化各因素的处理条件对最终的观测效果和结果都有影响,其中以甲基绿复染时间过长影响最大。同时,该染色法应用于贵妃鸡的T淋巴细胞计数中,4次重复的结果表明,贵妃鸡酸性α-醋酸萘酯酶阳性细胞率为37.86±1.89%,各重复间差异不显著。     

CYP450 epoxygenase/EET pathway attenuates adipose inflammation of obese mice through HIF-1α

AIM To investigate the effects of cytochrome P450 （CYP450） epoxygenase/epoxyeicosatrienoic acid （EET） pathway on insulin resistance in obese mice, and to explore the possible mechanisms. METHODS High-fat diet-induced obesity model was established in C57BL/6Cnc mice, and the obese mice were randomly divided into 3 groups, including obesity group （treated with saline; n=10）, EET group （treated with 11,12-EET; n=10） and EET inhibitor 14,15-epoxyeicosa-5（Z）-enoic acid （EEZE） group （n=10）. Normal C57BL/6Cnc mice （n=10） treated with saline served as control. Protein expression of CYP2J2 （one of CYP450 epoxygenases） and hypoxia-inducible factor-1α （HIF-1α） was measured by Western blot. Vessel-like structure was detected by immunofluorescence staining. The serum levels of insulin, tumor necrosis factor-α （TNF-α）, interleukin （IL）-1β, IL-6 and monocyte chemoattractant protein-1 （MCP-1） were measured by ELISA. RESULTS In obese mice, homeostasis model assessment of insulin resistance （HOMA-IR） values were increased, the protein level of CYP2J2 was reduced, and the protein level of HIF-1α was increased in adipose tissues as compared with the controls （P<0.05）. The serum levels of MCP-1, IL-1β, IL-6 and TNF-α were also significantly increased in obese mice （P<0.05）. After treatment with 11, 12-EET, the HOMA-IR values were decreased compared with vehicle-treated obese mice, HIF-1α expression levels were decreased in the adipose tissue, and the serum levels of MCP-1, IL-1β, IL-6 and TNF-α were reduced （P<0.05）. Immunohistochemical results of adipose tissue from vehicle-treated obese mice showed a marked decrease in vessel-like structures （CD31-positive） compared with normal control mice （P<0.05）. EET treatment significantly increased the newly formed vessel-like structures in the visceral adipose tissues of obese mice as compared with vehicle-treated obese mice （P<0.05）. CONCLUSION High-fat diet-induced obesity and insulin resistance are closely related to the CYP450 pathway. Exogenous EETs effectively decrease obesity-induced insulin resistance possibly through pro-angiogenesis and attenuation of hypoxia and inflammation.     

CYP11A1、CYP17A1、CYP19A1基因在小尾寒羊下丘脑-垂体-卵巢轴的表达

为揭示类固醇生成相关基因CYP11A1、CYP17A1、CYP19A1在绵羊多羔繁殖中的作用,以不同繁殖力小尾寒羊为研究对象,采用实时荧光定量PCR技术检测CYP11A1、CYP17A1、CYP19A1基因在小尾寒羊大脑、小脑、下丘脑、垂体、子宫、卵巢、输卵管中的表达。结果表明:CYP11A1基因主要在小尾寒羊卵巢、输卵管、子宫内表达;CYP11A1基因在单羔小尾寒羊卵巢的表达极显著低于多羔群体(P0.01),在子宫的表达量极显著高于多羔群体(P0.01);CYP17A1基因主要在小尾寒羊卵巢、输卵管、下丘脑表达;CYP17A1基因在多羔小尾寒羊群体下丘脑、子宫的表达量显著低于单羔群体(P0.05)。CYP19A1基因主要在小尾寒羊卵巢、下丘脑表达。本究结果提示CYP11A1、CYP17A1基因可能参与小尾寒羊多羔性状的调控。     

外源脱落酸和冠菌素对小麦籽粒萌芽的影响

[目的]通过研究外源脱落酸(ABA)和冠菌素(COR)对小麦籽粒萌芽的影响,探索COR用于防治小麦收获期穗发芽的可行性。[方法]以易穗发芽的白皮小麦品种济麦22和抗穗发芽的红皮小麦品种扬麦16为材料,以不同浓度ABA和COR处理两品种小麦籽粒后进行萌芽,测定萌芽率、芽长以及萌芽过程中的α-淀粉酶活性。[结果]38μmol/L以上浓度的ABA处理和0.05μmol/L以上浓度的COR处理均对两品种籽粒的萌发及芽的生长表现出显著的抑制作用,且COR抑制小麦种子萌发的生物活性约为ABA的200～1 500倍;浓度高于76μmol/L的ABA和0.05μmol/L的COR对两品种小麦籽粒萌发过程中的α-淀粉酶活性有抑制作用,且浓度越高抑制程度越大。[结论]ABA和COR可能通过抑制α-淀粉酶活性来抑制小麦籽粒的萌发和芽的生长;同浓度同一抑制剂处理下,白皮的济麦22萌芽及α-淀粉酶活性受抑制的程度显著高于扬麦16。该研究为COR用作控制大田小麦收获期穗发芽的可行性提供理论依据。