二维制备高效液相色谱分离塞隆骨强极性多肽北大核心CSCD

本实验建立了一种新型的利用高正交性的二维制备型高效液相色谱系统分离强极性动物药多肽的方法。本文以塞隆骨水提取物为研究对象,以亲水性C18AQ制备型高效色谱柱为第一维分离柱,首先在一维分离中将目标混合物分成若干组份;然后以C18MP制备型高效液相色谱柱为第二维色谱分离柱,将第一维分离后得到的组份纯化为单体化合物。本研究最终得到5个塞隆骨单体化合物,化合物纯度均超过98%。经Nano-LCESI-MS/MS鉴定和搜库分析,这些多肽的序列分别为:KTAILVKE、RGAPQDQE、LVGPGAPGR、GFAGD和KPQWHP。此研究方法速度快、效率高且重复性好,可以对类似的研究提供借鉴。     

胶原海绵的羟脯氨酸含量测定

选用普通实验室均能实现的 Woessner第 法对自制胶原海绵和 Gelfix(国外样品 )进行了羟脯氨酸含量测定 ,结果表明 ,该方法操作简单 ,重复性好 :自制胶原海绵的羟脯氨酸含量稳定 ,与国外样品的羟脯氨酸含量接近 ,且与胶原蛋白的羟脯氨酸含量接近 ,证实了两种胶原海绵的纯度均较高     

细胞壁的制备及其羟脯氨酸含量的测定

细胞壁结构和功能关系的研究越来越被人们所重视。伸展蛋白是植物细胞壁中一类重要的结构蛋白,与细胞壁的精细结构及各种生理功能密切相关。其特点之一是它的羟脯氨酸(Hyp)含量高达30％～40％。因此,细胞壁的制备及其羟脯氨酸含量的分析是细胞壁及其蛋白质研究中最基本又极重要的内容。本文介绍一种细胞壁的制备及其羟脯氨酸含量测定的简单方法。     

红光抑制绿豆下胚轴伸长时富含羟脯氨酸蛋白质的含量变化

红光能增加细胞壁的Ｈｙｐ含量和细胞各部分的Ｈｙｐ／Ｐｒｏ,绿豆下胚轴切段伸长被红光抑制的程度与两者呈正相关。环己酰亚胺和放线菌素Ｄ明显降低细胞壁的Ｈｙｐ含量,提高ＷＳＦＣ和ＣＷＲＳＥ中的Ｈｙｐ／Ｐｒｏ,抑制切段伸长。     

羟脯氨酸的研究与开发

从羟脯氨酸平衡失调与疾病、检测方法与应用、生产工艺、展望等4个方面介绍了羟脯氨酸的研究与开发。     

L-羟脯氨酸对小鼠生命活动的影响

用添加L 羟脯氨酸的饲料饲喂小鼠一个月后 ,对小鼠体重、抗缺氧、抗饥饿、抗旱、抗疲劳、耐热、耐寒等生理指标进行试验 ,并与对照组进行比较。结果表明 :L 羟脯氨酸有抑制体重增长的作用 ,可增加小鼠的抗缺氧能力、抗旱能力、抗疲劳能力、耐寒能力 ,对小鼠的生命活动有一定影响     

微等离子束对豚鼠皮肤超微结构分析和羟脯氨酸测定

目的观察微等离子束对豚鼠皮肤胶原组织作用效应的组织学和超微结构变化及羟脯氨酸含量测定,探讨微等离子束的作用机理。方法选择15只豚鼠,每只豚鼠背部划分为实验侧和空白对照侧2个等分区域,给予60W/10 kJ微等离子束照射,于作用后即刻、1周后和1月后分别切取实验侧及空白对照部位皮肤行组织病理维多利亚-立春红染色,透射电镜分析和羟脯氨酸检测试剂盒进行含量测定。结果 60 W/10 kJ即刻表现为表皮局灶性出现点阵状改变,部分表皮出现汽化缺失或者坏死变性,真皮浅层胶原组织出现点阵化表现和明显均质化;特殊染色显示微等离子束主要影响真皮胶原纤维,形成局灶性胶原纤维凝集和变性。1周后皮肤浅层胶原组织结构逐渐致密,排列有序,有少量组织细胞。1月后皮肤浅层胶原组织明显增厚,胶原纤维增粗并排列致密,弹力纤维呈局灶性增粗。透射电镜显示微等离子束作用后表皮细胞较完整,细胞间结构正常,但真皮胶原丧失正常结构,细胞结构消失,大量细胞凋亡明显,1月后仍可见少量细胞凋亡的表现但胶原结构逐渐恢复,浅层胶原纤维排列明显致密。羟脯氨酸测定显示微等离子束作用1周后羟脯氨酸含量要高于作用之前,但是差异性不具有统计学意义（P〉0.05）;1月后羟脯氨酸含量要明显高于作用前,差异性具有统计学意义（P〈0.05）。结论微等离子束对豚鼠皮肤胶原组织作用有明显的刺激效应,其主要靶组织为真皮胶原组织,可以明显促进皮肤新生胶原的增生。     

牛磺酸,羟脯氨酸,γ—氨基丁酸,鸟氨酸和色氨酸的测定

在日立８３５－５０型氨基酸分析仪上,用７２ｍｉｎ程序进行分析,通常一次只能分析１７种氨基酸。采用本文所述方法则一次能分析２２种氨基酸。在不延长分析时间,不增加成本的情况下,一次多分析了五种氨基酸,提高了仪器工作效率。     

595 nm脉冲染料激光非损伤嫩肤治疗对皮肤羟脯氨酸含量的影响

羟脯氨酸的含量是反映胶原纤维变化的敏感生化指标.本文利用昆明小鼠作为动物模型,研究不同能量的595 nm脉冲染料激光对皮肤羟脯氨酸含量的影响.20只昆明小鼠随机分为两组,1个实验组和1个对照组.三种能量的激光各照射5次,每次间隔3天,光斑有10%的重叠.照射后测皮肤羟脯氨酸含量的变化,SPSS进行数据分析. 8 J/cm2、10 J/cm2和12 J/cm2组羟脯氨酸含量分别比对照组增加39% 、50% 和58%(p<0.05),12 J/cm2能量组的效果最好,但他们之间没有统计学上的显著差异,表明595nm的脉冲染料激光照射能明显增加皮肤羟脯氨酸的含量,用于非损伤嫩肤治疗是有效的.     

红光抑制绿豆下胚轴伸长时富含羟脯氨酸蛋白质的含量变化

红光能增加细胞壁的Hyp含量和细胞各部分的Hyp／Pro,绿豆下胚轴切段伸长被红光抑制的程度与两者是正相关。环己酰亚胺和放线菌素D明显降低细胞壁的Hyp含量,提高WSFC和CWRSE中的Hyp／Pro,抑制切段伸长。香豆素不仅显著减少细胞壁的Hyp含量,而且降低WSFC和CWRSE中的Hyp／Pro,促进下胚轴伸长。α,α．二吡啶降低细胞壁的Hyp含量和WSFC中的Hyp／Pro,解除红光对切段伸长的抑制。EGTA、verapamil、La3 和A23187均可解除红光抑制切段伸长的作用,但对细胞壁的Hyp含量和细胞三部分蛋白质中Hyp／Pro影响较小。     

Hydroxyproline in Tetrahymena*

SYNOPSIS. Hydroxyproline (HP) can be quantified by the sensitive colorimetric procedure of Kivirikko et al. (1967). Without preliminary hydrolysis, only the free imino acid (I) can be detected. After hydrolysis in 6 N HCl at 120 C for 15 hr, also the peptide-bound (II) and polypeptide-bound (III) is detected. Cells grown in 2% (w/v) proteose peptone supplemented with 0.4% yeast (w/v) extract (PPY) contained 0.01 mg HP/mg cellular proteins. Over 95% was in the I or II form (soluble in cold 20% trichloro acetic acid). Cells grown in a chemically defined medium (DM), contained less than 0.34 μg/mg cellular proteins. Whereas the DM does not contain any HP, the PPY medium is rich in HP (0.032 mg/mg proteose peptone). In conclusion, the HP found in the cells grown on PPY is a “contaminant'’from this medium. No endogenous production of HP was demonstrated.     

Differentiation of Cancellous Bone and Medullary Bone in Laying Hens: a Novel Technique for Image Analysis

A selective staining technique for the identification and differentiation of cancellous bone from medullary bone of the laying hen by image analysis is described. Undecalcified Polymaster resin sections were oxidized in acidified potassium permanganate and oxalic acid before being immersed in an ammoniacal silver solution. The sections were reduced in formalin, fixed in sodium thiosulfate and counterstained in naphthalene black 10B which was dissolved in picric and acetic acids. Intensely stained cancellous bone was prominent with this technique compared with a paler medullary bone component which permitted the former to be easily recognized and measured by image analysis.     

红豆草抗羟脯氨酸变异系原生质体培养后愈伤组织的抗性特征

通过NaN3诱变得到的红豆草抗羟脯氨酸（Hyp）变异系,在酶液中游离原生质体进行培养,获得再生植株。在含不同浓度NaCl、羟脯氨酸或PEG的MS培养基上,原生质体来源抗性意伤组织中的游离脯氨酸含量在1周之内均急剧增加,随后开始下降,3周后接近正常水平。随着胁迫程度的提高,抗性愈伤组织中游离脯氨酸含量呈递增趋势,生长速度呈递减趋势。     

Validation of Bone Conversion in Osteoconductive and Osteoinductive Bone Substitutes

Bone reconstruction can be performed with an autogeneic graft from various donor regions. Osteoconductive and osteoinductive bone substitutes originate from substances of diverse chemical and morphological types and can have a synthetic or a biological derivation. Alongside autogeneic bone transplants and allogenic and xenogeneic bone implants, alloplastic bone replacements of synthetic or semi-synthetic origin are being used for defect reconstruction. In an animal model in rabbits five bone substitutes and one autogeneic graft were surgically incorporated into identical bone defects (10times 10 mm in size) in six anatomically defined regions of the skull. With scintigraphic and histological methods, the metabolic dynamics of the bone is examined as it reacts to the transplantation of autogeneic bone or to implanted bone replacement material. The different autogeneic, xenogeneic and alloplastic bone replacement materials can be differentiated according to the functional quality of the new tissue and the dynamics of the bone conversion thus induced. In the comparison of mineralized, osteoconductive bone subsitutes (TCP, HA, calcium carbonate ceramics) with demineralized, osteoinductive implants (DBM new, DBM old) and autogeneic bone grafts, the bone inducing matrices show the largest quantity of new bone formation, making possible a volume-constant reconstruction. This revised version was published online in July 2006 with corrections to the Cover Date.     

The Cytological Recognition of African Siderosis On Bone Smears

Eight cases of African siderosis primarily recognized on cytological smears are presented and discussed. All the smears were obtained from Jamshidi needle biopsies of vertebral bodies. Six cases showed siderosis only, while a seventh showed two pathological processes on one slide, namely metastatic keratinizing squamous carcinoma and siderosis. The remaining case showed cytological evidence of tuberculosis and siderosis. All cases were histologically confirmed, an additional feature in two cases being osseous tuberculosis which was not evident on the cytological smears. A search of the literature failed to reveal any report on the cytological recognition of this disease, or its association with tuberculosis.     

The Analysis of Bacterial Results in the Kidney Perfusion Fluid and Bone Allografts in Human Multiorgan Donors

In 19 young human multiorgan donors, we simultaneously analyzed the bacterial contamination of the kidney perfusion fluid and all retrieved bone allografts. Donor exclusion criteria were done according to the American and European Association of Tissue Banks excluding all patients with perforating wounds. The kidney perfusate revealed a contamination in 17 of 19 (89.5%) donors. Allograft testing demonstrated positive bacterial growth in 34 of 76 allografts (44.7%). Microorganisms originated from the normal skin flora and could be related to contamination during the harvesting procedure. In 5 cases we cultured identical bacterial subspecies in both cultures as a possible sign for systemic bacterial spreading during the multiorgan harvesting procedure.     

A Method for Histological Preparation of Undecalcified Bone Sections Containing Acrylic Bone Cement

An improved and time reducing method is presented for the histological evaluation of bone containing polymethylmethacrylate (PMMA) bone cement. The undecalcified bone was embedded in epoxy resin and sections of 50-100 μm thickness were produced using a commercially available cutting grinding system. The sections were stained with Stevenel's blue and van Gieson picrofuchsin or a modified hematoxylin-eosin. PMMA bone cement was not dissolved and remained enabling examination in situ of an intact cement bone interface and tissue reaction without decalcification.     

骨组织工程研究进展

本文主要介绍了骨组织工程学的概念以及研究内容,综述了骨组织工程的研究现状和发展方向。     

长骨缺损修复用生物医学材料

本文根据天然骨的组织和结构特点,论述了长骨缺损对修复材料的要求,提出了通过材料复合,仿生设计和快速成型制备功能梯度长骨修复材料的方法。