MV-mimicking micelles loaded with PEG-serine-ACP nanoparticles to achieve biomimetic intra/extra fibrillar mineralization of collagen in vitro

Since their discovery, matrix vesicles (MVs) containing minerals have received considerable attention for their role in the mineralization of bone, dentin and calcified cartilage. Additionally, MVs' association with collagen fibrils, which serve as the scaffold for calcification in the organic matrix, has been repeatedly highlighted. The primary purpose of the present study was to establish a MVs–mimicking model (PEG-S-ACP/micelle) in vitro for studying the exact mechanism of MVs-mediated extra/intra fibrillar mineralization of collagen in vivo. In this study, high-concentration serine was used to stabilize the amorphous calcium phosphate (S-ACP), which was subsequently mixed with polyethylene glycol (PEG) to form PEG-S-ACP nanoparticles. The nanoparticles were loaded in the polysorbate 80 micelle through a micelle self-assembly process in an aqueous environment. This MVs–mimicking model is referred to as the PEG-S-ACP/micelle model. By adjusting the pH and surface tension of the PEG-S-ACP/micelle, two forms of minerals (crystalline mineral nodules and ACP nanoparticles) were released to achieve the extrafibrillar and intrafibrillar mineralization, respectively. This in vitro mineralization process reproduced the mineral nodules mediating in vivo extrafibrillar mineralization and provided key insights into a possible mechanism of biomineralization by which in vivo intrafibrillar mineralization could be induced by ACP nanoparticles released from MVs. Also, the PEG-S-ACP/micelle model provides a promising methodology to prepare mineralized collagen scaffolds for repairing bone defects in bone tissue engineering.     

大鼠缰核与下丘脑外侧区在调节心血管活动方面的机能联系

电刺激大鼠下丘脑外侧区（LH）,动脉压明显升高,心率加快,在刺激电极同侧缰核（Hb)内微量注射盐酸利多卡因、电刺激LH引起的升压反应可被阻断38.9%,心率增快反应可被阻断44.4%,双侧Hb内微量注射盐酸利多卡因,电刺激LH引起的升压反应可被阻断40.7%,心率增快反应可被阻断41.2% ,单侧或双侧Hb内微量注射人工脑脊液均不能阻断电刺激LH引起的心血管反应。电刺激大鼠Hb,动脉压明显升高,心率无明显改变,在刺激电极同侧LH内微量注射盐酸利多卡因,电刺激Hb引起的升压反应可被阻断63.2%,双侧LH内微量注射盐酸利多卡因,电刺激Hb引起的升压反应可被阻断62.6%,单侧或双侧LH内微量注射人工脑脊液均不能阻断电刺激Hb引起的心血管反应。本实验提示Hb与LH在调节心血管活动方面有协同作用。     

Effects of chronic bifemelane hydrochloride administration on receptors for N-methyl-d-aspartate in the aged-rat brain

We assayed N-methyl-d-aspartate (NMDA) receptors [³H]3-(2-carboxypiperazin-4-yl)propyl-1-phosphonic acid ([³H]CPP) bindings) and evaluated their distribution in the brain by quantitative autoradiography in young adult and aged rats. In the young adult rats, NMDA receptors were present at relatively high concentrations in the cerebral cortex and hippocampus. In the aged rats, NMDA receptors were decreased in the nealy all areas of the brain, especially in the cerebral cortex and hippocampus. Chronic administration of bifemelane hydrochloride, a drug for sequela of cerebrovascular diseased, at a dose of 15 mg/kg/day for 14 days, markedly attenuated these decrease in NMDA receptors. Since NMDA receptors are considered to be involved in memory and learning processes, our results suggest that bifemelane hydrochloride may be applicable to the treatment of disturbed memory and learning.     

Thermodynamic stability of proteins in salt solutions: A comparison of the effectiveness of protein stabilizers

The denaturation of proteins by guanidine hydrochloride was studied in the presence of different concentrations of stabilizing salts, namely potassium phosphate, ammonium sulfate, and potassium acetate. The denaturation transition was followed by observing changes in the peptide circular dichroism atpH 7.0 and 25°C. From these results the free energy of stabilization for the process native denatured was determined. It was found that the stabilizing power of the anions increased in the order acetate < sulfate < phosphate, in agreement with the anionic lyotropic series. Ribonuclease A, which is known to have a site that can bind either a phosphate or a sulfate ion, showed a larger stabilization by these anions than that for lysozyme, pepsinogen, and myoglobin.     

Evidence for coexistence of ATP and nitric oxide in non-adrenergic,non-cholinergic (NANC) inhibitory neurones in the rat ileum,colon and anococcygeus muscle

The possible coexistence of the two non-adrenergic, non-cholinergic (NANC) inhibitory neurotransmitters, adenosine 5-triphosphate and nitric oxide in the myenteric plexus was investigated using whole-mount preparations of rat ileum, proximal colon and anococcygeus muscle. The presence of adenosine 5-triphosphate in neurones was examined using the quinacrine fluorescence technique. After localizing and taking photographs of quinacrine-fluorescent neurones and nerve fibres, the same tissues were then fixed and processed for NADPH-diaphorase activity, a marker for nitric oxide-containing neurones. We have demonstrated for the first time that almost all quinacrine-fluorescent myenteric neurones in the proximal colon are also NADPH-diaphorase reactive, while only a subpopulation of quinacrine-fluorescent neurones in ileum and anococcygeus muscle were also NADPH-diaphorase reactive.     

Chirospecific synthesis of D and Lp-chlorohomophenylalanine N-t-BOC DCHA salts

Summary The chirospecific conversions of D-glucosamine hydrochloride and D-mannosamine hydrochloride to the configurationally stable L and D isomers of N-t-butyloxycarbonylserinal were carried out byt-butylcarbonylation followed by sodium borohydride reduction and sodium meta-periodate oxidation. Reaction of the L and D aldehydes with the Wittig reagent prepared from 4-chlorobenzyltriphenylphosphonium chloride and butyl lithium followed by catalytic hydrogenation, Jones oxidation and salt formation with dicyclohexylamine gave the DCHA salts of the D and L isomers ofp-chlorohomophenylalanine N-t-Boc in high enatiomeric excess. The optical purity of the title compounds was established by hydrolysis to the respective free amino acids, followed by chiral derivatization and HPLC analysis.This was presented at the Fifth International Kyoto Conference on new Aspects of Organic Chemistry, Kyoto, Japan, November 11–15, 1991. Abstract #GO-13.     

盐酸艾司洛尔联合右美托咪定对肺癌患者应激反应和Th1/Th2型细胞因子的影响

摘要 目的：探讨盐酸艾司洛尔联合右美托咪定对肺癌患者应激反应和辅助性T细胞1（Th1）/Th2型细胞因子的影响。方法：选取武汉科技大学附属天佑医院2020年6月~2022年3月期间收治的肺癌根治术患者112例。按照随机数字表法将患者分为对照组（n=56,常规麻醉联合盐酸右美托咪定注射液）和研究组（n=56,对照组基础上联合盐酸艾司洛尔注射液）。对比两组苏醒质量、肺功能指标、应激反应指标、Th1/Th2型细胞因子和术后肺部并发症发生率变化情况。结果：研究组的呼吸恢复时间、定向力恢复时间、拔管时间短于对照组（P<0.05）。研究组术后肺部并发症发生率低于对照组（P<0.05）。研究组术毕动态肺顺应性（Cdyn）高于对照组,平台压（Pplat）、气道峰压（Ppeak）低于对照组（P<0.05）。研究组术毕皮质醇（Cor）、去甲肾上腺素（NE）及肾上腺素（E）低于对照组（P<0.05）。研究组术毕肿瘤坏死因子-α（TNF-α）、白细胞介素-2（IL-2）低于对照组（P<0.05）,研究组术毕白细胞介素-10（IL-10）、转化生长因子-β1（TGF-β1）高于对照组（P<0.05）。结论：盐酸艾司洛尔联合右美托咪定用于肺癌根治术中,具有较好的麻醉效果,可有效减轻术中应激反应,调节免疫功能并保护肺功能,还可降低术后肺部并发症发生率。     

单甲脒农药对模型池塘生态系统群落结构的影响

在３ｍ×１ｍ×１ｍ（Ｖ＝３ｍ３）含有底泥的模型池塘生态系统中研究单脒农药对水生生物群落结构的影晌。规定实验浓度力０、１．５、３．０、６．０和１２．０ｍｇ／Ｌ,每１５ｄ加入１次２５％单甲脒农药水剂,连续加入４次,实验进行２个多月。在实验浓度范围内,童甲脒农药对水生生物群落产生不同程度的影响。浮游生物比较敏感：加药后头几天内,种类、数量及多样性指数下降,浓度越大,影响越明显;大约１周以后,各处理组浮游生物群落逐步得到恢复,实验后期其数量甚至可超过对照水平,但群落结构发生改变,敏感种类少或消失,耐污种类增加,生物多样性降低。底栖生物比较耐污：处理槽大型水生植物的叶绿素含量有所减少,但其种类和生物量未见明显差异;底栖动物种类、数量也未见明显变化。微生物最耐污,在处理槽水层及沉积物中好氧异养菌数量有所增加,沉积物中厌气菌数量也有增加的趋势。青鱼对单甲脒农药较敏感,在１．５ｍｇ／Ｌ以下浓度尚能正常存活和繁殖。单甲脒农药水剂明显增加水体氮、磷含量,尤其磷酸盐含量高,使水体氮、磷比例失调,可能导致水体富营养化。根据综合指标分析,在规定单甲脒盆酸盐浓度＜１．５ｍｇ／Ｌ的实验条件下,水生生物群落结构未见明显改变．     

Novel function of guanidine hydrochloride in reverse micellar extraction of lysozyme from chicken egg white

The efficiency of guanidine hydrochloride (GuHCl) addition in the suppression of gel formation and the extraction of lysozyme during reverse micellar extraction from chicken egg white was investigated. A low concentration of GuHCl in the feed permitted the successful extraction of lysozyme in its native form without gel formation, which is perceived as a novel function of GuHCl. The highest recovery and specific activity of lysozyme were obtained at a GuHCl concentration of 0.06 M in 25 mM AOT reverse micellar extraction from 20-fold-diluted natural chicken egg white. Lysozyme and ovalbumin CD spectra in the corresponding GuHCl aqueous solutions revealed no changes in the higher order structures of the proteins. Furthermore, the specific activity of lysozyme in the feed was well preserved in the GuHCl system. (c) 1995 John Wiley & Sons, Inc.     

Investigation of the folding pathway of the TEM-1 β-lactamase

The TEM-1 β-lactamase is a globular protein containing 12 proline residues. The folding mechanism of this enzyme was investigated by kinetic and equilibrium experiments with the help of fluorescence spectroscopy and circular dichroism. The equilibrium denaturation of the protein induced by guanidine hydrochloride occurs in two discrete steps, indicating the existence of a thermodynamically stable intermediate state. Thisstate is 5.2 ± 0.4 kcal/mol less stable than the native conformation and 5.7 ± 0.2 kcal/mol more stable than the fully denaturedprotein. This intermediate state exhibits a high content of native secondary structure elements but is devoid of specific tertiary organization; its relation to the “molten globule” is discussed. Refolding kinetic experimentsrevealed the existence of a transient intermediate conformation between thethermodynamically stable intermediate and the native protein. This transient intermediate appears rapidly during the folding reaction. It exhibits a secondary structure content very similar to that of the native protein and has also recovered a significant amount of tertiary organisation. The final refolding step of the TEM-1 β-lactamase, leading to the native enzyme, is dominated by two major slow kinetic phases which probablyreflect a very complex process kinetically limited by proline cis/transisomerization. © 1995 Wiley-Liss, Inc.