PPAR‐β facilitating maturation of hepatic‐like tissue derived from mouse embryonic stem cells accompanied by mitochondriogenesis and membrane potential retention

Relatively little is known about mitochondria metabolism in differentiating embryonic stem (ES) cells. Present research focused on several elements of cellular energy metabolism in hepatic‐like tissue derived from mouse ES cells. We demonstrated that mitochondrial location patterns and mitochondrial membrane potential (ΔΨ_m) existed in subsequent differentiation of the tissue. Mitochondriogenesis appeared at the early stage and kept a normal ΔΨ_m in differentiated mature hepatocytes. Peroxisome proliferator‐activated receptor‐α (PPAR‐α) expression was transitorily increased at the beginning, and kept a relatively low level later, which accompanied by expression of PPAR‐γ coactivator (PGC)‐1α, a master regulator of mitochondrial biogenesis. PPAR‐β expression showed robust up‐regulation in the late differentiation course. Enhanced co‐expressions of PPAR‐β and albumin with catalysis of UDP‐glucuronosyltransferases (UGTs) were observed at mature stage. While PPAR‐γ expression changed little before and after differentiation. Mitochondriogenesis could be accelerated by PPAR‐α specific agonist WY14643 and abolished by its antagonist GW6471 at the early stage. Neither of them affected mitochondrial ΔΨ_m and albumin generation in the differentiated hepatocytes. Furthermore, maturation of hepatic‐like tissue and mitochondriogenesis in hepatocyte could be efficiently stimulated by PPAR‐β specific agonist L165041 and abolished by PPAR‐β specific antagonist GSK0660, but not affected by PPAR‐γ specific agonist GW1929. In conclusion, the derived hepatic tissue morphologically possessed cellular energy metabolism features. PPAR‐α seemed only necessary for early mitochondriogenesis, while less important for ΔΨ_m retention in the mature tissue derived. The stimulation of PPAR‐β but not ‐γ enhanced hepatogenesis, hepatocytes maturation, and mitochondriogenesis. PPAR‐β took an important role in cellular energy metabolism of hepatogenesis. J. Cell. Biochem. 109: 498–508, 2010. © 2009 Wiley‐Liss, Inc.     

Dammaranes from Gynostemma pentaphyllum and synthesis of their derivatives as inhibitors of protein tyrosine phosphatase 1B

Protein tyrosine phosphatase 1B (PTP1B) is a key factor in the negative regulation of insulin pathway and a promising target for treatment of diabetes and obesity. Herein, the sapogenin 2b, prepared from the natural triterpene saponin 1b, was modified at 3-position to establish the dammarane derivatives library via esterification, oxidation and reductive amination reaction and evaluated as PTP1B inhibitors. 3-O-para-Carboxylphenyl substituted derivative 5b was found with the best in vitro inhibition activity to protein tyrosine phosphatase 1B (IC₅₀ = 0.27 μM), where 3-O-meta-carboxylphenyl substituted 5a exhibited the best selectivity (nearly fivefolds) between PTP1B and T-cell protein tyrosine phosphatase.     

Conjugating folic acid to gold nanoparticles through glutathione for targeting and detecting cancer cells

Gold nanoparticles (GNPs) were modified with glutathione (GSH) to form GSH-capped GNPs, which have carboxyl groups on the surface of these nanoparticles. Then folic acid (FA) was conjugated with GNPs through the reaction between amino group of FA and carboxyl group of GSH. These folic acid-conjugated nanoparticles (FA-GSH-GNPs) were stable in aqueous solution over a broad range of pH and ionic strength values. The targeting of FA-GSH-GNPs in human cervices carcinoma cells (HeLa cells) with high-level folate receptor expression was confirmed by transmission electron microscopy (TEM) and confocal laser scanning microscopy (CLSM). No cellular uptake of these nanoparticles was observed in A549 cells lack of folate receptor. HeLa cells and mouse fibroblasts incubated with FA-GSH-GNPs were assayed by measuring the relative absorbance of the supernatant collected at low-speed centrifugation. Based on this simple spectroscopic method, HeLa cells have been detected with a detection limit of 10² cells/mL.     

乳源活性肽对抗猪瘟病毒抗体IgG、IgA和IgM水平的影响

不同剂量乳源活性肽与2头份猪瘟活疫苗同时分点肌肉注射仔猪,每周一次耳静脉采血,间接ELISA法检测抗猪瘟病毒抗体IgG、IgA和IgM水平.发现乳源活性肽能够明显提高抗猪瘟病毒抗体水平.当乳源活性肽为15 g/L时,与对照组相比,IgG抗体水平在注射后28 d时提高了21.0%,IgA和IgM抗体水平在注射后14 d时分别提高了13.8%和7.8%.实验组腹泻率、发病率和死亡率明显比对照组低,同时发现乳源活性肽具有促进生长的作用.     

Development of HBsAg-Binding Aptamers that bind HepG2.2.15 cells via HBV surface antigen

Hepatitis B virus surface antigen(HBsAg),a specific antigen on the membrane of Hepatitis B virus (HBV)-infected cells,provides a perfect target for therapeutic drugs.The development of reagents with high affinity and specificity to the HBsAg is of great significance to the early-stage diagnosis and treatment of HBV infection.Herein,we report the selection of RNA aptamers that can specifically bind to HBsAg protein and HBsAg-positive hepatocytes.One high affinity aptamer,HBs-A22,was isolated from an initial ...     

Cd2+ 、Zn2+单一及复合胁迫对凤眼莲根质膜3种氧化还原酶活性的影响

采用水培法研究了不同浓度Cd2+ 、Zn2+单一及复合胁迫(0.05和0.50 mmol·L-1 Cd2+,0.5和5.0 mmol·L-1 Zn2+)对凤眼莲[Eichhornia crassipes (Mart. ) Solms]幼苗根质膜3种氧化还原酶[NADH氧化酶、Fe(CN)63-还原酶和EDTA-Fe3+还原酶]活性的影响. 结果表明, Cd2+ 、 Zn2+单一及复合胁迫对凤眼莲根质膜NADH氧化酶、Fe(CN)63-还原酶及EDTA-Fe3+还原酶活性的影响效应有明显的差异.0.05或0.50 mmol·L-1 Cd2+单一胁迫处理20 h可使凤眼莲根质膜3种氧化还原酶活性均较对照显著降低(P≤0.05),0.5或5.0 mmol·L-1 Zn2+单一胁迫20 h可导致凤眼莲根质膜NADH氧化酶和EDTA-Fe3+还原酶活性也均较对照显著降低(P≤0.05).随胁迫时间的延长,Cd2+ 、 Zn2+单一及复合胁迫处理均可使凤眼莲根质膜氧化还原酶活性增强,胁迫处理20 d时凤眼莲根质膜3种氧化还原酶活性均高于胁迫20 h的活性.Cd2+-Zn2+复合胁迫对凤眼莲根质膜3种氧化还原酶活性的影响效应因作用时间和胁迫浓度的不同而有一定的差异;在胁迫20 h时,Cd2+与Zn2+之间对凤眼莲根质膜3种氧化还原酶活性的作用关系较复杂,因胁迫浓度的不同表现出协同或拮抗的关系;胁迫20 d时,Cd2+与Zn2+对凤眼莲根质膜3种氧化还原酶活性的复合影响表现出明显的拮抗关系.研究结果显示,Cd2+ 、 Zn2+对凤眼莲根质膜氧化还原酶的复合作用效应与Cd2+和Zn2+的浓度比例及胁迫时间均相关.     

长江中游地区野生百合资源调查及利用前景

通过对长江中游地区野生百合资源进行调查,发现长江中游地区有野生百合资源14个种和3个变种即野百合(Lilium brownii F. E. Brown ex Miellez var. brownii)、百合(L. brownii F. E. Brown ex Miellez var. viridulum Baker)、宜昌百合(L. leucanthum (Baker) Baker)、渥丹(L. concolor Salisb. var. concolor)、有斑百合(L. concolor Sali sb. var. pulchellum (Fisch.)Regel)、滇百合(L. baberianum Coll.et Hemsl)、大理百合(L.taliense Franch.)、药百合(L.speciosum Thunb. var. gloriosoides Baker)、湖北百合(L.henryi Baker)、南川百合(L. rosthornii Diels)、宝兴百合(L. duchartrei Franch.)、山丹(L. pumilum DC.)、川百合(L. davidii Duchartre)、条叶百合(L. callosum Sieb. et Zucc.)、乳头百合(L. papilliferum Franch.)、绿花百合(L. fargesii Franch.)和卷丹(L. tigrinum Ker Gawler)。对野生百合在长江中游地区分布现状、观赏特性及开发利用价值进行了分析,为百合新品种培育奠定基础。同时对长江中游地区野生百合资源的保护和合理开发利用提出建议。     

TRPM7 Activates m-Calpain by Stress-Dependent Stimulation of p38 MAPK and c-Jun N-Terminal Kinase

TRPM7 is a Ca²⁺-permeant and Mg²⁺-permeant ion channel in possession of its own kinase domain. In a previous study, we showed that overexpression of the channel-kinase in HEK-293 cells produced cell rounding and loss of adhesion, which was dependent on the Ca²⁺-dependent protease m-calpain. The TRPM7-elicited change in cell morphology was channel-dependent and occurred without any significant increase in cytosolic Ca²⁺. Here we demonstrate that overexpression of TRPM7 increased levels of cellular reactive oxygen species (ROS) and nitric oxide, causing the activation of p38 mitogen-activated protein kinase (MAPK) and c-Jun N-terminal kinase (JNK). Application of inhibitors of p38 MAPK and JNK blocked TRPM7-induced cell rounding and activation of m-calpain, without affecting the phosphorylation state of the protease. Overexpression of TRPM7 increased intracellular Mg²⁺; however, when the concentration of either external Ca²⁺ or Mg²⁺ was increased to favor the permeation of one divalent cation over the other, a similar increase in cell rounding and calpain activity was detected, indicating that TRPM7-mediated activation of m-calpain is not dependent on the nature of the divalent conducted by the channel. Application of inhibitors of nitric oxide synthase and mitochondrial-derived ROS reduced TRPM7-induced increases in nitric oxide and ROS production, blocked the change in cell morphology, and reduced cellular calpain activity. Collectively, our data reveal that excessive TRPM7 channel activity causes oxidative and nitrosative stresses, producing cell rounding mediated by p38 MAPK/JNK-dependent activation of m-calpain.     

VOLATILE CONSTITUENTS OF CHAROPHYTES AS OVIPOSITION DETERRENTS OF CULEX PIPIENS PALLENS (DIPTERA: CULICIDAE)1

Four populations of charophytes (including three species), Chara inconnexa T. F. Allen (populations 1 and 2), C. vulgaris L., and Nitellopsis obtusa (Desv.) J. Groves, were studied for effective chemicals as oviposition deterrents of Culex pipiens pallens. The charophyte volatile organic compounds (VOCs) were retained in Tenax GR, subsequently desorbed using a thermal desorption cold trap injector (TCT), and analyzed by gas chromatography/mass spectrometry (GC/MS) to elucidate that charophytes have repellent properties. C. inconnexa (1) and C. inconnexa (2) exhibited strong repellent activities, and C. vulgaris showed some repellent activity against C. pipiens pallens with terpenes and benzothiazole playing major roles, while N. obtusa lacked those compounds and did not have an effect. These results suggest that charophytes have potential application as pesticides, but there are interspecific differences. In addition, benzene hydrocarbons were among the volatiles in Chara but not in N. obtusa, implying that some charophytes could be used to absorb these compounds.     

Endothelium oriented differentiation of bone marrow mesenchymal stem cells under chemical and mechanical stimulations

Bone marrow mesenchymal stem cells (MSCs) have multi-differentiation capability. Their endothelial cell (EC) oriented differentiation is the key to vasculogenesis, in which both mechanical and chemical stimulations play important roles. Most previous studies reported individual effects of VEGF or fluid shear stress (SS), when MSCs were subjected to shear stress of 10–15 dyn/cm² over 24 hr. In this paper, we investigated responses of MSCs from young Sprague Dawley rats to shear stress, VEGF and the combination of the two stimuli. Our study showed that the combined stimulation of shear stress and VEGF resulted in more profound EC oriented differentiation of MSCs in comparison to any individual stimulation. Furthermore, we subjected MSCs to prolonged period of fluid shear stimulation, i.e. 48 hr rather than 24 hr, and increased the magnitude of the shear stress from 10 dyn/cm² to 15, 20 and 25 dyn/cm². We found that without VEGF, the endothelium oriented differentiation of MSCs that was seen following 24 hr of shear stimulation was largely abolished if we extended the shear stimulation to 48 hr. A similar sharp decrease in MSC differentiation was also observed when the magnitude of the shear stress was increased from 10–15 dyn/cm² to 20–25 dyn/cm² in 24 hr shear stimulation studies. However, with combined VEGF and fluid shear stimulation, most of the endothelial differentiation was retained following an extended period, i.e. at 48 hr, of shear stimulation. Our study demonstrates that chemical and mechanical stimulations work together in determining MSC differentiation dynamics.