Mash2基因在体细胞克隆牛肺脏中DNA甲基化状态分析

体细胞核移植(体细胞克隆)技术在动物生产、医药工业、治疗性克隆以及对珍稀濒危动物的拯救有重要意义,然而克隆效率低下以及克隆动物发育异常,严重制约了克隆技术的发展和应用．在体细胞核克隆中,供体核来自高度分化了的体细胞,发生在核移植后几小时内供体核的重编程,决定了克隆胚胎的发育能力．印记基因是由等位基因表观遗传修饰的不对称导致的基因表达具有亲本选择性,而DNA甲基化是调控印记的一个主要方式．印记基因Mash2在胚胎发育和器官形成过程中起着非常重要的作用．为了探求核移植过程中Mash2基因DNA 甲基化的表观重编程是否充分,利用亚硫酸氢盐测序法对出生48 h内死亡的体细胞核移植牛和正常对照牛肺脏中Mash2基因的DNA甲基化状态进行分析．结果显示,尽管位于Mash2基因启动子和第一个外显子处的CpG岛在正常牛和克隆牛中甲基化水平都不高(20.04%,5.55%),但克隆组的甲基化水平仍显著低于正常对照组 (P < 0.05)．甲基化模式正常组中9N3有5种不同的形式,9N4仅1种;而克隆组9C3和9C5也分别是1种．推测Mash2基因的异常DNA甲基化很可能是导致克隆牛肺脏发育异常的一个重要原因．     

Phenolic compounds and their anti-oxidative properties and protein kinase inhibition from the Chinese mangrove plant Laguncularia racemosa

Phenolic compounds, named integracin D (1), (7′R, 8′S, 8S)-8-hydroxyisoguaiacin (3), (2R, 3R) pinobanksin-3-caffeoylate (5) and threo-8S-7-methoxysyringylglycerol (6), respectively, were isolated from the Chinese mangrove plant Laguncularia racemosa (L) Gaertn. f. (Combretaceae), together with 23 known phenolic metabolites. Their structures were elucidated on the basis of extensive spectroscopic analyses including that of IR, UV, MS, CD, 1D and 2D NMR spectra as well as by comparison with literature data. Compound 5 showed significant anti-oxidative activity in the DPPH and TEAC free-radical-scavenging assays, while several of the phenolic compounds were tested for protein kinase inhibitory activity in an assay involving 24 different human tumor related protein kinases. Compounds 5, 7, and 23 showed potential inhibition with IC₅₀ values between 2.2 and 3.6 μg/mL toward individual kinases. The ellagic acid derivatives were tested for insecticidal activity.     

益坤宁对围绝经期大鼠卵巢细胞凋亡率及caspase-3表达的影响

目的:研究中药益坤宁(yikunning,YKN)对围绝经期大鼠卵巢细胞凋亡率及凋亡相关基因caspase-3基因表达的影响,探讨益坤宁治疗围绝经期综合征的作用机理。方法:选用30只自然衰老的围绝经期雌性大鼠,随机分为中药益坤宁实验组、围绝经期对照组和利维爱(livial)对照组,另选10只青年雌性大鼠作为青年对照组。连续灌胃处理4周后,采用原位脱氧核糖核苷酸末端转移酶介导的缺口末端标记(TUNEL)法检测大鼠卵巢细胞凋亡率,采用逆转录-聚合酶链反应(RT-PCR)和蛋白印迹(Western blot)检测大鼠卵巢中caspase-3 mRNA和蛋白表达。结果:益坤宁组大鼠卵巢细胞凋亡率显著低于围绝经期对照组(P0.01);益坤宁组大鼠卵巢中caspase-3 mRNA和蛋白表达低于围绝经期对照组,高于青年对照组,差异有统计学意义(P0.01)。结论:中药益坤宁通过降低围绝经期大鼠卵巢细胞凋亡率,下调卵巢中凋亡相关基因caspase-3的表达,从而延缓卵巢衰老,这可能是其治疗围绝经期综合征的分子机制之一。     

Testin protects against cardiac hypertrophy by targeting a calcineurin‐dependent signalling pathway

Multiple organs express testin (TES), including the heart. Nevertheless, current understanding of the influence of TES on cardiovascular diseases, especially on cardiac hypertrophy and its etiology, is insufficient. This study investigated the influence of TES on cardiac hypertrophy and its etiology. Murine models with excessive TES expression specific to the heart were constructed with an adeno‐associated virus expression system. Cardiac hypertrophy was stimulated through aortic banding (AB). The severity of cardiac hypertrophy was evaluated through molecular, echocardiographic, pathological, and hemodynamic examination. The findings of our study revealed that TES expression was remarkably suppressed not only in failing human hearts but also in mouse hearts with cardiac hypertrophy. It was discovered that excessive TES expression driven by an adeno‐associated viral vector noticeably inhibited hypertrophy triggered by angiotensin II (Ang II) in cultivated cardiomyocytes from newborn rats. It was also revealed that TES knockdown via AdshTES caused the reverse phenotype in cardiomyocytes. Furthermore, it was proved that excessive TES expression attenuated the ventricular dilation, cardiac hypertrophy, dysfunction, and fibrosis triggered by AB in mice. It was discovered that TES directly interacted with calcineurin and suppressed its downstream signalling pathway. Moreover, the inactivation of calcineurin with cyclosporin A greatly offset the exacerbated hypertrophic response triggered by AB in TES knockdown mice. Overall, the findings of our study suggest that TES serves as a crucial regulator of the hypertrophic reaction by hindering the calcineurin‐dependent pathway in the heart.     

Enantiomeric Polyketides from the Starfish‐Derived Symbiotic Fungus Penicillium sp. GGF16‐1‐2

One new racemic mixture, penicilliode A ( 1 ) and four pairs of enantiomeric polyketides, penicilliode B and C ( 2 and 3 ) and coniochaetone B and C ( 4 and 5 ), were obtained from the starfish‐derived symbiotic fungus Penicillium sp. GGF16‐1‐2. Interestingly, the strain GGF16‐1‐2 can produce enantiomers. The absolute configuration of 1 was determined by X‐ray diffraction (XRD) analysis, and the absolute configurations of 2 – 4 were determined by the optical rotation (OR) values and electronic circular dichroism (ECD) calculations. Compounds 1 – 5 were firstly isolated from the marine‐derived fungus Penicillium as racemates, and 2 – 5 were separated by HPLC with a chiral stationary phase. All the compounds were evaluated for their antibacterial, cytotoxic and inhibitory activities against PDE4D2.     

Facile Patterning of Laser‐Induced Graphene with Tailored Li Nucleation Kinetics for Stable Lithium‐Metal Batteries

A facile and scalable approach is reported to stabilize the lithium‐metal anode by regulating the Li nucleation and deposition kinetics with laser‐induced graphene (LIG). By processing polyimide (PI) films on copper foils with a laser, a 3D‐hierarchical composite material is constructed, consisting of a highly conductive copper substrate, a pillared array of flexible PI, and most importantly, porous LIG on the walls of the PI pillars. The high number of defects and heteroatoms present in LIG significantly lowers the Li nucleation barrier compared to the copper foil. An overpotential‐free Li nucleation process is identified at current densities lower than 0.2 mA cm^?2. Theoretical computations reveal that the defects serve as nucleation centers during the heterogeneous nucleation of lithium. By adopting such composites, ultrastable lithium‐metal anodes are obtained with high Coulombic efficiencies of ≈99%. Full lithium‐metal cells based on LiFePO₄ cathodes with a material loading of ≈15 mg cm^?2 and a negative/positive ratio of 5/1 could be cycled over 250 times with a capacity loss of less than 10%. The current work highlights the importance of nucleation kinetics on the stability of metallic anodes and demonstrates a practical method toward long lasting Li‐metal batteries.     

Lipopolysaccharide-induced proliferation and glycolysis in airway smooth muscle cells via activation of Drp1

Abnormal airway smooth muscle cells (ASMCs) proliferation is an important pathological process in airway remodeling contributes to increased mortality in asthma. Mitochondrial dynamics and metabolism have a central role in the maintenance of the cell function. In this study, lipopolysaccharide (LPS)-induced ASMCs proliferative model was used to investigate the effect of mitochondria on the proliferation of ASMCs and the possible mechanism. We used cell and molecular biology to determine the effect of dynamin-related protein 1 (Drp1) on LPS-mediated ASMCs cell cycle progression and glycolysis. The major findings of the current study are as follows: LPS promoted an increased mitochondrial fission and phosphorylation of Drp1 at Ser616 (p-Drp1 Ser616). LPS-induced ASMCs proliferation and cell cycle progression, which was significantly inhibited application of Drp1 RNA interfering. Glycolysis inhibitor 2-deoxyglucose (2-DG) depressed ASMCs proliferative process induced by LPS stimulation. LPS caused mitochondrial metabolism disorders and aerobic glycolysis in a dependent on Drp1 activation. These results indicated that Drp1 may function as a key factor in asthma airway remodeling by mediating ASMC proliferation and cell cycle acceleration through an effect on mitochondrial metabolic disturbance.     

Recombinant neuritin affects the senescence,apoptosis, proliferation,and migration of rat bone marrow-derived mesenchymal stem cells

Objective

To study the effects of recombinant neuritin expressed by Pichia pastoris GS115 on the senescence, apoptosis, proliferation, and migration associated with rat bone marrow-derived mesenchymal stem cells (BMSCs).

Results

Recombinant neuritin was purified by Ni-affinity chromatography and identified by western blot and MALDI-TOF spectrometry. The effects of recombinant neuritin on senescence, apoptosis, proliferation, and migration of rat BMSCs WERE investigated. β-Galactosidase staining indicated that recombinant neuritin administration significantly inhibited BMSCs senescence at 1 μg neuritin/ml. Additionally, recombinant neuritin reduced the number of apoptotic cells at the early stage according to Annexin V/propidium iodide staining and inhibited cell proliferation according to MTT assay results. Moreover wound healing assay results showed that recombinant neuritin promoted BMSCs migration in the neuritin-treatment group.

Conclusion

Recombinant neuritin affects the senescence, apoptosis, proliferation, migration of rat BMSCs. Our findings offer insight into neuritin function outside of the nervous system.