Nicorandil prevents right ventricular remodeling by inhibiting apoptosis and lowering pressure overload in rats with pulmonary arterial hypertension

Background

Most of the deaths among patients with severe pulmonary arterial hypertension (PAH) are caused by progressive right ventricular (RV) pathological remodeling, dysfunction, and failure. Nicorandil can inhibit the development of PAH by reducing pulmonary artery pressure and RV hypertrophy. However, whether nicorandil can inhibit apoptosis in RV cardiomyocytes and prevent RV remodeling has been unclear.

Methodology/Principal Findings

RV remodeling was induced in rats by intraperitoneal injection of monocrotaline (MCT). RV systolic pressure (RVSP) was measured at the end of each week after MCT injection. Blood samples were drawn for brain natriuretic peptide (BNP) ELISA analysis. The hearts were excised for histopathological, ultrastructural, immunohistochemical, and Western blotting analyses. The MCT-injected rats exhibited greater mortality and less weight gain and showed significantly increased RVSP and RV hypertrophy during the second week. These worsened during the third week. MCT injection for three weeks caused pathological RV remodeling, characterized by hypertrophy, fibrosis, dysfunction, and RV mitochondrial impairment, as indicated by increased levels of apoptosis. Nicorandil improved survival, weight gain, and RV function, ameliorated RV pressure overload, and prevented maladaptive RV remodeling in PAH rats. Nicorandil also reduced the number of apoptotic cardiomyocytes, with a concomitant increase in Bcl-2/Bax ratio. 5-hydroxydecanoate (5-HD) reversed these beneficial effects of nicorandil in MCT-injected rats.

Conclusions/Significance

Nicorandil inhibits PAH-induced RV remodeling in rats not only by reducing RV pressure overload but also by inhibiting apoptosis in cardiomyocytes through the activation of mitochondrial ATP-sensitive K⁺ (mitoK_ATP) channels. The use of a mitoK_ATP channel opener such as nicorandil for PAH-associated RV remodeling and dysfunction may represent a new therapeutic strategy for the amelioration of RV remodeling during the early stages of PAH.     

脂蛋白电子显微结构研究的优化负染方法

人类心血管等疾病与胆固醇含量的高低有着十分密切的关系．人体内胆固醇主要通过血液中的脂蛋白来调节和运载．因此,新一代调节胆固醇的药物设计迫切地需要揭示脂蛋白分子结构与功能之间的关系．由于脂蛋白分子的成分高度复杂、结构特征灵活、尺寸微小,传统的蛋白质结构标定技术,如X射线衍射、核磁共振谱、质谱和冷冻电子显微镜等手段都面临着巨大的困难．针对这一问题,任罡小组最近提出一种优化的电镜负染制备方法,可以观测到单个脂蛋白分子的空间结构．此方法应用的成功,使得对单个脂蛋白分子的结构研究成为可能．同时也证明该负染色技术作为一种普遍的实验手段,可以用于对蛋白质小分子的结构研究,从而将促进新一代蛋白质分子药物的研发．     

Simple-MSSM: a simple and efficient method for simultaneous multi-site saturation mutagenesis

Objective

To develop a practically simple and robust multi-site saturation mutagenesis (MSSM) method that enables simultaneously recombination of amino acid positions for focused mutant library generation.

Results

A general restriction enzyme-free and ligase-free MSSM method (Simple-MSSM) based on prolonged overlap extension PCR (POE-PCR) and Simple Cloning techniques. As a proof of principle of Simple-MSSM, the gene of eGFP (enhanced green fluorescent protein) was used as a template gene for simultaneous mutagenesis of five codons. Forty-eight randomly selected clones were sequenced. Sequencing revealed that all the 48 clones showed at least one mutant codon (mutation efficiency = 100%), and 46 out of the 48 clones had mutations at all the five codons. The obtained diversities at these five codons are 27, 24, 26, 26 and 22, respectively, which correspond to 84, 75, 81, 81, 69% of the theoretical diversity offered by NNK-degeneration (32 codons; NNK, K = T or G).

Conclusion

The enzyme-free Simple-MSSM method can simultaneously and efficiently saturate five codons within one day, and therefore avoid missing interactions between residues in interacting amino acid networks.

     

通过共转化和花药培养快速获得直链淀粉含量降低且无抗性标记的转基因水稻

用根癌农杆菌共转化的方法将p13W8质粒(含反义蜡质基因)和pCAMBIA1300质粒(含潮霉素抗性基因)导入水稻.经PCR检测,发现在29个T1群体中发生抗潮霉素抗性基因和反义蜡质基因的分离;从1 264个T1单株中筛选到183个只带反义蜡质基因片段的转基因植株.选择了4个直链淀粉含量降低的T1单株进行花药培养,获得正常结实的花培苗34株;经PCR检测,有23株为只含反义蜡质基因而无潮霉素抗性基因的植株.从转化到获得直链淀粉含量降低、遗传稳定的转基因植株仅用了一年半时间.     

枸杞瘿螨姬小蜂的生物学特性

枸杞瘿螨姬小蜂Cirrospilus eniophyesiYang(新种)是枸杞瘿螨Aceria pallidaKeifer的重要天敌,以幼虫在虫瘿内取食枸杞瘿螨的成虫和若虫。在(25±1)℃下,该蜂完成1个世代需16~21d。成虫寿命及产卵量与补充营养有关,取食10%蜂蜜水的雌虫寿命为(6.94±0.67)d,雄虫为(5.14±0.49)d。不进行补充营养的雌雄成虫寿命分别为(1.96±0.27)d和(1.06±0.29)d;补充10%蜂蜜水和水可显著提高产卵量,每雌平均产卵量分别为(29.13±6.12)粒和(23.14±3.16)粒,产卵高峰期在羽化后的第2天。成虫羽化时间多集中在6:00~8:00am和4:00~6:00pm,羽化当天即可交尾。枸杞瘿螨姬小蜂自然种群消长与寄主虫口密度相关,在6月底7月初寄生率达到83.3%。     

杨黑点叶蜂幼虫龄数和龄期

夏季,采用室内常温、常光照条件下饲养的方法和解剖镜测微技术,研究杨黑点叶蜂Pristophora conjugata Dahlbom幼虫龄数和龄期。结果表明,杨黑点叶蜂雄性幼虫共5龄,雌性幼虫共6龄;各龄幼虫头宽值分别为(mm):0.585~0.849;0.849~1.371;1.371~1.960;1.960~2.350;2.350~2.868;2.868。根据幼虫头宽值确定幼虫所处龄期,为准确掌握防治时期提供科学依据。     

四种植物精油成分对枸杞棉蚜的生物活性

【目的】研究植物精油4种成分丁香酚(eugenol)、α-松油醇(α-terpineol)、肉桂油(cinnamon oil)和香茅油(citronella oil)对枸杞棉蚜Aphis gossypii的生物活性以及马来酸二乙酯和水杨酸乙酯对这些化合物对枸杞棉蚜毒力的影响,为无公害防治枸杞(Lycium spp.)害虫奠定科学基础。【方法】采用二项选择法测定上述4种精油成分(1%)对枸杞棉蚜无翅成蚜的驱避作用;玻璃管药膜法进行生物测定,评估这4种化合物及其与马来酸二乙酯或水杨酸乙酯的混合液对枸杞棉蚜成蚜的毒力;采用生物化学方法分析这些化合物处理后枸杞棉蚜成蚜乙酰胆碱酯酶(AChE)和三磷酸腺苷酶(ATPase)的离体活性。【结果】α-松油醇(1%)对枸杞棉蚜无翅成蚜的驱避作用明显,1, 2和3 h的驱避率分别为58.1%, 58.6%和55.7%; 3 h时测试的4种化合物对枸杞棉蚜成蚜的驱避率由大到小依次为α-松油醇>香茅油>肉桂油>丁香酚。丁香酚对枸杞棉蚜成蚜的毒杀作用最强,处理4 h时的LC₅₀值为0.48 mL/L,肉桂油、α-松油醇、香茅油对枸杞棉蚜成蚜的LC₅₀值分别为1.23, 6.24和11.97 mL/L。对枸杞棉蚜成蚜的生物测定结果表明,马来酸二乙酯对香茅油、肉桂油、丁香酚和α-松油醇的增效比分别为2.75, 2.86, 1.78和1.56;水杨酸乙酯对香茅油、肉桂油、丁香酚和α-松油醇的增效比分别为3.41, 2.67, 2.00和1.89。毒力比结果表明,所测定的4种化合物对枸杞棉蚜的毒力相差49.9倍,丁香酚与马来酸二乙酯或水杨酸乙酯的混合液对枸杞棉蚜的触杀毒力最高。4种精油成分对枸杞棉蚜成蚜AChE和ATPase活性有不同的作用,丁香酚对AChE活性的抑制作用最强,但对ATPase活性表现为低浓度促进,高浓度抑制。【结论】α-松油醇对枸杞棉蚜驱避作用最好;丁香酚、α-松油醇、肉桂油和香茅油这4种精油成分对枸杞棉蚜的毒力相差很大;丁香酚与马来酸二乙酯或水杨酸乙酯混合后对枸杞棉蚜的毒力提高,具有进一步开发应用的潜力;所测精油成分对枸杞棉蚜离体AChE和ATPase活性有一定抑制作用。     

萝卜块根中两个具溶菌酶活性的几丁质结合蛋白的纯化及其特性

通过亲和层析和羧甲基一纤维素离子交换层析从萝卜的块根中分离到两个具溶菌酶活性的酶组份：CBP1和CBP2。两者经SDS-PAGE均显示单一蛋白染色条带,其对应的分子量分别为26．9kD和24．8kD。两种蛋白除有溶菌酶活性外,还有几丁质酶活性,但无壳聚糖酶活性。各种类型的几丁质对CBP1和CBP2都有较强的吸附作用,而在还原／非还原的单向SDS-PAGE中却观察不到两者分子中存在二硫键。     

SD-PMA-ddPCR检测食品中单核细胞增生李斯特氏菌

【目的】检测食品中单核细胞增生李斯特氏菌活菌。【方法】利用脱氧胆酸钠(SD)对受损细胞预处理,然后使叠氮溴化丙锭(PMA)进入受损细胞与DNA发生共价交联,提取细菌基因组DNA进行微滴式数字PCR(dd PCR)检测。【结果】0.1%SD和5.0 mg/L PMA协同作用,可以有效抑制108 CFU/m L的单核细胞增生李斯特氏菌死菌DNA的PCR扩增。经过SD和PMA对样品预处理,dd PCR可以在死菌存在条件下,定量检测鸡肉中单核细胞增生李斯特氏菌活菌,消除了"假阳性"结果的出现。活菌灵敏度检测结果显示:SD-PMA-dd PCR的灵敏度为2.0 copies/20μL。SD-PMA-dd PCR方法精密度和稳定性良好。【结论】SD-PMA-dd PCR在检测食源性致病菌方面有巨大的发展空间。     

落叶松尺蠖Erannis ankeraria Staudinger的生物学特性

【目的】明确落叶松尺蠖Erannis ankeraria Staudinger的发生规律及生物学特性。【方法】野外调查和室内饲养。【结果】在集宁市落叶松尺蠖1年1代,以卵越冬,越冬卵在翌年4月下旬、5月上旬孵化,6月中下旬开始入土化蛹,成虫于9月初羽化、产卵。在温度为20℃,RH=70%的条件下,落叶松尺蠖幼虫期(19.96±0.86)d,预蛹期(3.93±0.95)d,蛹期(108.4±13.17)d。蛹分布于树干基部30~90cm范围内,深度4~8 cm。雄虫在6:00—8:00及12:00—16:00羽化,早于雌性,雌虫在20:00—24:00羽化,羽化若干小时后可交尾,交配持续时间20~260 min,可多次交尾。20℃下交尾雌虫寿命为(5.56±1.47)d,雄虫为(3.95±0.95)d,产卵量为(162.2±69.9)粒;不交尾雌性为(8.03±2.90)d,雄性为(4.38±1.59)d,产卵量为(164.1±81.3)粒,但卵不能孵化;15℃条件下不交尾雌雄寿命分别为(14.48±6.67)d,(6.64±1.76)d,产卵量为(145.7±76.8)粒。【结论】落叶松尺蠖危害期短,蛹期较长,雌雄成虫羽化时间有差异;温度和交尾对寿命和产卵量都有显著影响。