Extracellular matrix components direct porcine muscle stem cell behavior

In muscle tissue, extracellular matrix proteins, together with the vasculature system, muscle-residence cells and muscle fibers, create the niche for muscle stem cells. The niche is important in controlling proliferation and directing differentiation of muscle stem cells to sustain muscle tissue. Mimicking the extracellular muscle environment improves tools exploring the behavior of primary muscle cells. Optimizing cell culture conditions to maintain muscle commitment is important in stem cell-based studies concerning toxicology screening, ex vivo skeletal muscle tissue engineering and in the enhancement of clinical efficiency. We used the muscle extracellular matrix proteins collagen type I, fibronectin, laminin, and also gelatin and Matrigel as surface coatings of tissue culture plastic to resemble the muscle extracellular matrix. Several important factors that determine myogenic commitment of the primary muscle cells were characterized by quantitative real-time RT-PCR and immunofluorescence. Adhesion of high PAX7 expressing satellite cells was improved if the cells were cultured on fibronectin or laminin coatings. Cells cultured on Matrigel and laminin coatings showed dominant integrin expression levels and exhibited an activated Wnt pathway. Under these conditions both stem cell proliferation and myogenic differentiation capacity were superior if compared to cells cultured on collagen type I, fibronectin and gelatin. In conclusion, Matrigel and laminin are the preferred coatings to sustain the proliferation and myogenic differentiation capacity of the primary porcine muscle stem cells, when cells are removed from their natural environment for in vitro culture.     

Effects of Cystamine on antioxidant activities and regulatory T cells in lupus‐prone mice

Attenuated antioxidant activities, irregular cytokines expressions and reduced regulatory T cells, are strongly associated with the pathogenesis of systemic lupus erythematosus (SLE). Despite the well‐established beneficial effects of cystamine on lupus‐prone mice, the extent to which cystamine contributes to antioxidant activity and the reduction of regulatory T cells has seldom been investigated. Therefore, this study elucidates how cystamine affects anti‐oxidant activities in NZB/W F1 mice by performing assays of Glutathione (GSH), 1,1‐diphenyl‐2‐ picryl‐hydrazyl (DPPH) and malondialdehyde thiobarbituric acid (MDA). In addition, investigations of the effects of cystamine on CD4⁺/CD25⁺ regulatory T cells and interleukin‐6 (IL6)/STAT‐3 signalling were performed with flow cytometry and immunoblots. Experimental results reveal more significantly reduced MDA and increased GSH and DPPH in NZB/W F1 mice receiving cystamine than in those mice receiving PBS. Meanwhile, CD4⁺/CD25⁺ regulatory T cells more significantly increase in NZB/W F1 mice receiving cystamine than in those mice receiving PBS, accompanied by significantly reduced IL‐6/phosphorylated STAT‐3 expression. The above findings suggest the beneficial effects of cystamine in terms of increasing antioxidant activities and CD4⁺/CD25⁺ regulatory T cells in lupus‐prone mice by suppressing IL‐6/STAT3 signalling.     

经皮穴位电刺激联合常规镇痛对无痛人流术后宫缩痛镇痛效果及机制研究

摘要 目的：探讨经皮穴位电刺激联合常规镇痛对无痛人流术后宫缩痛镇痛效果及机制。方法：选择来我院行无痛人流患者100例。根据随机数字表法分为两组。对照组行术后常规镇痛治疗,同时在内关、合谷、三阴交最酸胀处黏贴电极但是不进行穴位刺激。观察组在术后常规镇痛基础上行经皮穴位电刺激治疗。对比不同时间点两组患者的机体一般情况、不同时间点两组患者的宫缩痛疼痛情况、T3时正性负性情绪量表评分、T3时血清P物质、前列腺素、5-羟色胺、β-内啡肽、强啡肽水平、T4点时的心理状态评分及满意度评分。结果：对照组T1、T2点时的BP、HR、RR、SpO₂明显较同组T0点低,对照组T3点与观察组T1、T2、T3点时BP、HR、RR、SpO₂与同组T0点对比无统计学意义（P>0.05）;对照组T1、T2点时的BP、HR、RR、SpO₂明显较观察组同时间点低（P<0.05）。T0点,两组VAS评分对比无统计学意义（P>0.05）;T1、T2、T3点时间,观察组的VAS评分明显较对照组低（P<0.05）;对照组T1、T2点的VAS评分明显较同组T0点高,观察组T1、T2、T3点的VAS评分明显较T0点低（P<0.05）。观察组的正性情绪明显较对照组高,负性情绪明显较对照组低（P<0.05）。观察组的血清P物质、前列腺素、5-羟色胺、β-内啡肽、强啡肽水平明显较对照组低（P<0.05）。观察组的心理状态评分明显较对照组低,满意度评分明显较对照组高（P<0.05）。结论：经皮穴位电刺激联合常规镇痛可明显改善无痛人流术后宫缩痛镇痛效果、患者心理状态及满意度,可能与其可降低血清P物质、前列腺素、5-羟色胺、β-内啡肽、强啡肽水平有关。     

Lymph node colonization induces tumor-immune tolerance to promote distant metastasis

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Liquid chromatographic separation of chiral diarylcarbinols

The direct HPLC separation of three chiral carbinols of general formula Mesityl-CH(OH)-Aryl has been achieved using Pirkle (R)-DNBPG ionic or covalent columns and, for Aryl = o-tolyl, on a Chiralpak OP(+) phase. It is apparent that steric hindrance and hydrogen bonding play important roles in chiral recognition. Two compounds structurally very similar but lacking the hydroxyl group were not resolved in their enantiomeric pairs. © 1992 Wiley-Liss, Inc.     

Substrate-enzyme interactions and catalytic mechanism in phospholipase C: a molecular modeling study using the GRID program.

Based on the high-resolution X-ray crystallographic structure of phospholipase C from Bacillus cereus, the orientation of the phosphatidylcholine substrate in the active site of the enzyme is proposed. The proposal is based on extensive calculations using the GRID program and molecular mechanics geometry relaxations. The substrate model has been constructed by successively placing phosphate, choline and diacylglycerol moieties in the positions indicated from GRID calculations. On the basis of the resulting orientation of a complete phosphatidylcholine molecule, we propose a mechanism for the hydrolysis of the substrate.     

虎杖象甲培植真菌营养特性及对菌丝附属物形成的影响

虎杖象甲培植共生真菌形成的共生体系是植菌昆虫菌业中的典型代表。共生真菌Penicillium herquei如何向虎杖象甲Euops chinensis提供营养尚未明确。本研究发现共生真菌P. herquei的菌丝表面存在大量瘤状凸起物及由凸起物衍生的附属丝等特化结构,该结构可能为虎杖象甲提供营养;对共生真菌的营养研究表明,共生真菌能高效利用山梨醇、蔗糖、海藻糖、葡萄糖等单糖或双糖,以及酪氨酸、甘氨酸、谷氨酰胺等昆虫非必须氨基酸,同时在高碳和最适碳源条件下有利于菌丝特化附属物的产生。研究结果不仅提供了植菌卷叶象甲菌业中共生真菌在营养方面的适应性进化证据,而且为进一步揭示共生真菌适应卷叶象甲的营养机制奠定了基础。     

Chlamydomonas reinhardtii-derived multimer Mytichitin-CB possesses potent antibacterial properties

Mytichitin-CB was isolated from Mytilus coruscus in 2014. This antimicrobial peptide shows a weak inhibitory effect on Gram-negative bacteria but inhibits the growth of Gram-positive bacteria and fungi efficiently. Here, a C-terminal hemagglutinin and 6×Histidine (HA-6×His) double tagged three tandem repeats of Mytichitin-CB (3×Mytichitin-CB) with a molecular weight of about 21.5 kDa was expressed in Chlamydomonas reinhardtii. The recombinant 3×Mytichitin-CB was stably expressed following continuous sixth passages of cells and inhibited the growth of both Gram-negative and Gram-positive bacteria at maximum inhibitory concentration (MIC) values between 30 and 50 μg/mL. 3×Mytichitin-CB was stable in terms of its antibacterial activity when treated by a wide range of temperatures and pHs and was resistant to digestion by various proteases. C. reinhardtii-derived 3×Mytichitin-CB had low hemolytic activity and cell cytotoxicity. Moreover, 3×Mytichitin-CB efficiently caused changes on the cell morphology by destroying membrane integrity of the tested bacteria. Our data thus, for the first time, show that C. reinhardtii is a suitable host for stably expressing recombinant 3×Mytichitin-CB, which possesses potent antibacterial properties.     

Dynamic control of toxic natural product biosynthesis by an artificial regulatory circuit

To mimic the delicately regulated metabolism in nature for improved efficiency, artificial and customized regulatory components for dynamically controlling metabolic networks in multiple layers are essential in laboratory engineering. For this purpose, a novel regulatory component for controlling vanillin biosynthetic pathway was developed through directed evolution, which was responsive to both the product vanillin and substrate ferulic acid, with different capacities. This regulatory component facilitated pathway expression via dynamic control of the intracellular substrate and product concentrations. As vanillin is an antimicrobial compound, low pathway expression and vanillin formation levels enabled better cell growth at an early stage, and the product feedback-activated pathway expression at later stages significantly improved biosynthesis efficiency. This novel multiple-layer dynamic control was demonstrated effective in managing the trade-off between cell growth and production, leading to improved cell growth and vanillin production compared to the conventional or quorum-sensing promoter-controlled pathway. The multiple-layer dynamic control enabled by designed regulatory components responsive to multiple signals shows potential for wide applications in addition to the dynamic controls based on biosynthetic intermediate sensing and quorum sensing reported to date.     

阴道微生态在早产中的研究进展

阴道微生态是由阴道的局部解剖结构、周期性的内分泌变化、阴道局部免疫系统和阴道内微生物菌群共同组成的阴道环境和生态系统。多项研究证实阴道微生态失调通过局部炎症因子释放、黏膜免疫应答的改变和局部代谢变化,可引起早产的发生。阴道内小分子物质如糖类、短链脂肪酸和胺类通过代谢路径和代谢产物在阴道微生态失调与早产的发病机制起作用。近年来,阴道微生态的理念得到重视,治疗方法由传统的抗生素治疗转向了综合治疗,目的是恢复正常阴道菌群和阴道上皮黏膜免疫系统。但阴道微生态与宿主之间存在复杂的相互作用,因此仍需要进一步的研究。