MK2 plays an important role for the increased vascular permeability that follows thermal injury

We previously reported Rho kinase is involved in vessel hyper-permeability caused by burns. Here we further explore the Rho kinase downstream signaling, it is found that its specific inhibitor Y27632 significantly diminishes the activation of JNK and p38 MAPKs but not ERK that induced by serum from burned rats (burn-serum). JNK activation was found involved in the expression of HUVEC adhesion molecules following thermal injury, although not in the process of stress fiber formation. Inhibition of various MAPKs by specific inhibitors showed that SB203580 (inhibitor of p38), but neither SP600125 (inhibitor of JNK) nor PD98059 (inhibitor of ERK), abolish activation of the p38 downstream kinase MK2. Demonstration of stress fibers by fluorescent-labeled phalloidin showed that inhibition of MK2, either by its specific inhibitor or by dominant negative adeno-viral-carried constructs, significantly reduced burn-serum-induced HUVEC stress-fiber formation, while inhibition of another downstream p38 MAPK kinase, PRAK, had no such effects. Transfection of dominant negative adeno-viral MK2 (Ad-MK2(A)) significantly inhibited thermal injury-induced blood vessel hyper-permeability in rats and, moreover, prolonged the survival of burned rats beyond 72 h following thermal injury. One of the mechanisms behind these phenomena is that Ad-MK2(A) causes a significant depression of burn-serum-induced HSP27-phosphorylation, while the adeno-viral transported dominant negative PRAK (Ad-PRAK(A)) does not block. Although the effect of blockade of MK2 through its adeno-viral approach requires further study and investigation of alternatives to know for sure, we may have found a new pathway behind thermal-injury-induced blood vessel hyper-permeability, namely: Rho kinase > p38 > MK2 > HSP27.     

AMPK在胆汁酸代谢中的调节作用

腺苷酸活化蛋白激酶的激活可以抑制胆固醇、脂肪的合成,最近研究表明,腺苷酸活化蛋白激酶的激活也可抑制胆汁酸合成及促进胆盐输出泵的生成,说明腺苷酸活化蛋白激酶在调节胆汁酸代谢中起着重要作用.本文将从腺苷酸活化蛋白激酶激活途径、胆汁酸代谢、腺苷酸活化蛋白激酶活性对胆汁酸合成及转运等方面进行综述.     

First-generation neuronal precursors in the crayfish brain are not self-renewing

Adult-born neurons in crayfish (Procambarus clarkii) are the progeny of 1st-generation precursor cells (functionally analogous to neuronal stem cells in vertebrates) that are located in a neurogenic niche on the ventral surface of the brain. The daughters of these precursor cells migrate along the processes of bipolar niche cells to proliferation zones in the cell clusters where the somata of the olfactory interneurons reside. Here they divide again, producing offspring that differentiate into olfactory local and projection neurons. The features of this neuronal assembly line, and the fact that it continues to function when the brain is isolated and perfused or maintained in organotypic culture, provide opportunities unavailable in other organisms to explore the sequence of cellular and molecular events leading to the production of new neurons in adult brains. Further, we have determined that the 1st-generation precursor cells are not a self-renewing population, and that the niche is, nevertheless, not depleted as the animals grow and age. We conclude, therefore, that the niche is not a closed system and that there must be an extrinsic source of neuronal stem cells. Based on in vitro studies demonstrating that cells extracted from the hemolymph are attracted to the niche, as well as the intimate relationship between the niche and vasculature, we hypothesize that the hematopoietic system is a likely source of these cells.     

High throughput ranking of recombinant avian scFv antibody fragments from crude lysates using the Biacore A100

Advances in molecular evolution strategies have made it possible to identify antibodies with exquisite specificities and also to fine-tune their biophysical properties for practically any specified application. Depending on the desired function, antibody/antigen interactions can be long-lived or short-lived and, therefore, particular attention is needed when seeking to identify antibodies with specific reaction-rate and affinity properties. Surface plasmon resonance (SPR) biosensors routinely generate sensitive and reliable kinetic data from antibody/antigen interactions for both therapeutic and diagnostic applications. However, many kinetic-based screening assays require rigorous sample preparation and purification prior to analysis. To ameliorate this problem, we developed a rapid and reliable assay for characterising recombinant scFv antibody fragments, directly from crude bacterial lysates. Ninety-six scFv antibodies derived from chickens immunised with C-reactive protein (CRP) were selected by phage display and evaluated using the Biacore A100 protein interaction array system. Antibodies were captured from crude bacterial extracts on the sensor chip surface and ranked based on the percentage of the complex left (% left) after dissociation in buffer. Kinetic rate constants (k(a) and k(d)) and affinity (K(D)) data were obtained for six clones that bound monomeric CRP across a broad affinity range (2.54 x 10(-8) to 3.53 x 10(-10) M). Using this assay format the A100 biosensor yielded high quality kinetic data, permitting the screening of nearly 400 antibody clones per day.     

The Selection of a Pharmaceutical Salt—The Effect of the Acidity of the Counterion on Its Solubility and Potential Biopharmaceutical Performance

A roadmap for the selection of a pharmaceutical salt form for a development candidate is presented. The free base of the candidate did not have sufficient chemical stability for development. The initially selected salt form turned out to be undevelopable because it was unstable during scale-up synthesis and storage. The rationale for the new solid form screening and the criteria for selection are discussed. Before the final selection, the pH solubility profiles of the 2 new salts, a benzoate and a besylate, were compared. Atypical solubility behavior was observed for the benzoate salt in hydrochloric acid with and without normal saline. A scheme is proposed illustrating how the pKas of the counterion and active pharmaceutical ingredient, the medium composition, and final pH affect the solubility and solution equilibria of the 2 selected salt forms. This scheme also includes the equilibria between solution and solid phases in different pH ranges. The pharmaceutical importance of this research is that it sheds light on how the acidity of the counterion can affect the solubility of the selected salt form in the gastric environment. With a well-designed formulation strategy, this property potentially can be translated to optimal biopharmaceutical performance of the drug product.     

杜仲饮片性状检定的探讨

目的：探讨市场中未去粗皮的杜仲饮片性状检定问题。方法：靶向性的收集10批样品（包括杜仲饮片与盐杜仲）,归纳切制形状的特点,按《中国药典2015年一部》检验;并对样品进行5种重金属及有害元素的检测;对其中6批未不合格Pb元素样品进一步分析粗皮与刮去粗皮饮片。结果：市场流通的饮片按切制形状主要分为4种类型,即竹帘状杜仲饮片、未去粗皮的块片状杜仲饮片,丝条状盐杜仲、未去粗皮的块片状盐杜仲。6批粗皮中重金属Pb元素明显高于相应刮去粗皮饮片的量。结论：未去粗皮杜仲饮片未按标准规定在加工时刮去粗皮,而粗皮中Pb元素检测值高,未去粗皮的杜仲饮片性状判定为不合格具有合理性。     

自制双层盐袋固定袋应用于永久起搏器植入术后压迫的效果分析

摘要 目的：探讨自制双层盐袋固定袋在永久起搏器植入术后患者切口压迫中的效果。方法： 2015年1月～2016年6月间在心内科行永久起搏器植入术的患者,随机进行分组。对照组患者术后使用两个500g普通盐袋压迫12～24 h,观察组患者术后采用自制的双层盐袋固定袋压迫切口6～12 h。观察患者术后并发症、心理、睡眠状况、舒适度,患者和护士对压迫工具的满意度。结果：观察组与对照组相比,观察组患者术后并发症少、患者心理、睡眠状况及术后舒适度更好,患者和护士的满意度高（P＜0.05）。结论：使用自制的双层盐袋固定袋压迫切口处,临床使用效果满意,有效降低术后并发症,提高患者的睡眠质量。 关键词 盐袋;永久起搏器;切口压迫;并发症;舒适度     

急性腹泻患者对口服补液盐认知水平调查及护理干预研究

目的了解急性腹泻患者对口服补液盐（ORS）的认知水平、选择意愿及使用情况，探讨护理干预措施。方法自行设计调查问卷对137例急性腹泻病住院患者进行调查，评估患者对ORS的认知水平、选择意愿；给予护理干预措施后．再次调查，比较护理干预前后患者对ORS认知水平的异同。结果本组患者对ORS的认知水平、选择意愿及正确服用率普遍较低；文化程度、患腹泻病次数、曾接触过ORS信息宣教等因素与ORS认知水平和选择意愿呈正相关。护理干预后，患者对ORS的认知情况明显改善，平均得分由（7．28±5．02）上升到（11．15±4．08）（t=7．00，P=0．000）；对ORS的选用率由原来的13．86％上升到72．26％（x^2=95．26，P=0．000）；ORS正确服用率由原来的7．29％升高到68．61％（x^2=109．35．P=0．000）。结论护理干预措施能提高腹泻患者对ORS的认知水平、选择意愿和正确使用率。     

The relationship between DNA adduct formation by benzo[a]pyrene and expression of its activation enzyme cytochrome P450 1A1 in rat

Benzo[a]pyrene (BaP) is a human carcinogen requiring metabolic activation prior to reaction with DNA. Cytochrome P450 (CYP) 1A1 is the most important hepatic and intestinal enzyme in both BaP activation and detoxification. CYP1A2 is also capable of oxidizing BaP, but to a lesser extent. The induction of CYP1A1/2 by BaP and/or β-naphthoflavone in liver and small intestine of rats was investigated. Both BaP and β-naphthoflavone induced CYP1A expression and increased enzyme activities in both organs. Moreover, the induction of CYP1A enzyme activities resulted in an increase in formation of BaP–DNA adducts detected by ³²P-postlabeling in rat liver and in the distal part of small intestine in vivo. The increases in CYP1A enzyme activity were also associated with bioactivation of BaP and elevated BaP–DNA adduct levels in ex vivo incubations of microsomes of both organs with DNA and BaP. These findings indicate a stimulating effect of both compounds on BaP-induced carcinogenesis.