Proline with or without Hydroxyproline Influences Collagen Concentration and Regulates Prolyl 4-Hydroxylase α(I) Gene Expression in Juvenile Turbot(Scophthalmus maximus L.)

This study was conducted to investigate the effect of dietary proline(Pro), and Pro and hydroxyproline(Hyp) in combination on the growth performance, total Hyp and collagen concentrations of tissues, and prolyl 4-hydroxylase α(I)(P4H α(I)) gene expression in juvenile turbot feeding high plant protein diets. A diet containing 50% crude protein and 12% crude lipid was formulated as the basal and control, on which other two protein and lipid contents identical experimental diets were formulated by supplementing the basal with either 0.75% Pro(Pro-0.75) or 0.75% Pro and 0.75% Hyp(Pro+Hyp). Four groups of fish in indoor seawater recirculating systems, 35 individuals each, were fed twice a day to apparent satiation for 10 weeks. The results showed that dietary Pro and Hyp supplementation had no significant effect on growth performance and feed utilization of juvenile turbot(P > 0.05). Total Hyp and collagen concentrations in muscle were significantly increased when dietary Pro and Hyp increased(P < 0.05), and fish fed diet Pro+Hyp showed significantly higher free Hyp content in plasma than those fed other diets(P < 0.05). The expression of P4 H α(I) gene in liver and muscle was significantly up regulated in fish fed diet Pro-0.75 in comparison with control(P < 0.05); however the gene was significantly down regulated in fish fed diet Pro+Hyp in muscle in comparison with fish fed diet Pro-0.75(P < 0.05). It can be concluded that supplement of crystal L-Pro and L-Hyp to high plant protein diets did not show positive effects on growth performance of juvenile turbot, but enhanced total collagen concentrations in muscle.

Peripheral Concentration of Hydroxyproline and 3-methyl-histidine in Postpartum Cow

A total of 16 Friesian cows were used in the study to determine the peripheral concentration of hydroxyproline(HYPRO) and 3 methyl histidine(3 MEHIS).The objectives of the experiment were to find out the relationship between collagen and myosin degradation and uterine involution in the postpartum cow.The results showed tht the mean concentrations of HYPRO and 3 MEHIS were 13.47±2.38 and 18.35±2.77 n mole/mL,respectively,for the cows during the days 60～90 postpartum.And for the cows immediately after calving the concentrations of HYPRO and 3 HEHIS reduced from 34.29 and 28.06 nmole/mL on the day one after calving to the normal lever of 13.23 and 17.97 n mole on the day 50 postpartum,respectively.There was a good correlation between the peripheral concentrations of both HYPRO and 3 MEHIS and the diameters of the involuting uterus(r=0.8128).

Urinary Concentrations of Hydroxyproline and 3-Methyl Histidine in Postpartum Cow

The urinary concentrations of hydroxyproline (HYPRO)and 3 -methyl histidine (3 - MEHIS) were de- termined in 16 Chinese-Holstein cows. The objectives of the experiment were to find out thhe relationship between collagen and myosin ddegradation and uterine involution in the postpartum cow. The results in the experiment showed that the mean concentrations of HYPRO and 3-MEHIS were 138.32±22.99 and 37.09 ±3.90 nmol·mL-1,respectively,for the cows during the days between 60-90 postpartum,and for the cows immediately after calving the concentrations of HYPRO and 3 -MEHIS reduced from 284.30 and 65.48 nmol ·mL-1 on the day one after calving to the normal level of 109.18 and 33.51 nmol·mL^-1 on the day 50 post- partum,respectively. There was a good correlation between the urinary concentrations of both HYPRO and 3 -MEHIS and the diameters of the involuting uterus (r = 0. 79).

Plant regeneration from protoplasts of hydroxyprolineresistant cell line in Onobrychis viciaefolia

An efficient protocol for plant regeneration from protoplasts of hydroxyproline(HYP)resistant cell line of Onoblychis viciaefolia was established. In SH medium supplemented with 1 mg/L 2, 4-dichlorophenoxy-acetic acid (2,4-D), 0.5 mg/L kinetin (KT) and 0.2 mg/L naphthalene acetic acid (NAA), the division frequency of protoplastderived cells reached uP to over 60 %, and microcalli were obtained in 5-6 wk. Upon transferring them on agar solidified MS medium plus 2 mg/L indole-3-acetic acid (IAA), shoots were induced. After cultivating them on MS medium with or without IAA, roots were regenerated.Chromosome number of all protoplast-regenerated plants examined were normal (2n=28). The protoplast-derived calli and plants grew vigorously on the medium containing 10 mmol/L HYP.

CHANGES OF URINARY HYDROXYPROLINE IN FRACTURE PATIENTS

Fifteen cases of fresh fractures have been studied, including 14 cases of tubu-lar bone fractures with age from 28 to 60 years and one case of vertebral fracture ofdorsolumbar region, 16 years of age. There were 13 male and 2 female patients. Theyhad no endocrine disturbances or other bone diseases. The ratio ofhydroxyproline/creatinine (HP/Cr) in these cases increased gradually within 20-30 daysafter fracture and then reached the peak value which was about 3-4 times of normal, thepeak value maintained for a long time. If the trauma was mild or the patient was young,the peak began to decline on the 60th day after fracture. While in those severe cases withprolonged confinement to bed, the peak only began to come down about 110th dayafter fracture. The ratio of HP/Cr decreased rapidly from the peak at first, then pursueda slow course. Generally, the ratio of HP/Cr returned to normal on the 240th to 310thday. Bone union began at a time when peak value started to decline, and with completionof the remodelling process, the HP/Cr value returned to normal.

Correlation between Vitamin C Deficiency and Hydroxyproline Amino Acid in Young Children of Northern Part in Palestine

Vitamin C is a water soluble vitamin found in many natural sources, citric fruits, vegetables, particularly as antioxidants and as a factor to catabolism and metabolism. The aim of this article is to correlate the level of vitamin C and the hydroxyproline level in urine samples among young palestinian children from age 5 - 14 years. Materials and Methods: Urine samples of 34 individuals of both sexes, their ages ranged from 5 - 14 years were collected and analyzed for determination of Urinary Hydroxyproline by using Modified Neuman and Logan Method. The result of this study shows a significant correlation between Vitamin C and Hydroxyproline at level 0.01. The conclusion of this study was that low vitamin C intake was found among young palestinian children from age 5 - 14 years, Vitamin C should be supplemented in the drinks taken by young palestinian children from age 5 - 14 years as well as further research and investigation with large samples required to include all children from middle and southern Palestine.

清热生肌膏介导NLRP3对放射性皮炎大鼠MPO、Hyp表达的影响

目的 清热生肌膏介导NLRP3炎性小体对放射性皮炎大鼠皮肤损伤修复及MPO、Hyp表达水平的影响。方法 选取SD健康大鼠90只,80只随机分为正常组、模型组(放射性皮炎大鼠模型)、治疗组(模型+清热生肌膏)、对照组(模型+复方茶多酚软膏),每组各20只。另设抑制剂组(NLRP3抑制剂)10只。通过创面愈合率衡量创面愈合情况。透射电镜观察创面细胞超微结构。HE染色观察病理学变化。分光光度比色法测定MPO活性。免疫印迹检测NLRP3、Caspase-1蛋白表达。结果 各组在治疗后第1 d时的创面愈合无差异(P>0.05)。模型组大鼠在治疗3~21 d时的创面愈合均低于治疗组及对照组(P<0.05)。治疗组与对照组大鼠治疗治疗3~21 d各时间点创面愈合比较无差异(P>0.05)。与正常组相比,模型组大鼠MPO表达升高、Hyp表达降低(P<0.05)。与模型组相比,治疗组大鼠MPO表达降低、Hyp表达升高(P<0.05)。治疗组与对照组MPO及Hyp表达对比无差异(P>0.05)。与正常组相比,模型组大鼠NLRP3、Caspase-1表达升高(P<0.05)。与模型组相比,治疗组大鼠NLRP3、Caspase-1蛋白表达降低(P<0.05)。治疗组与抑制剂组大鼠NLRP3、Caspase-1蛋白表达对比无差异(P>0.05)。结论 清热生肌膏可抑制MPO水平并促进Hyp表达,对放射性皮炎大鼠皮肤损伤修复有积极促进作用,其作用机制可能与介导NLRP3炎性小体有关。

Assay for Hydroxyproline of Collagen Sponge

The content of hydroxyproline of collagen sponge is tested by Woessner Method I.The hydroxyproline standard curve and regression equation are described.The content of hydroxyproline of collagen sponge is tested subsequently.The result show that the hydroxyproline content is consistent with that of collagen protein,which could be concluded that the purity of collagen sponge,either prepared in our institute or that from abroad is high.

Synthesis and Characterization of a Novel Functional Biodegradable Copolymer-Poly(lactic acid-4-hydroxyproline-polyethylene glycol)

A series of poly（lactic acid-4-hydroxyproline-polyethylene glycol） （PLA-Hpr-PEG） copolymers were synthesized by direct melt copolymerization of D,L-lactic acid and 4-hydroxyproline with different feed amount of polyethylene glycol （PEG） 0.1%, 0.5%, 1% and 5%, respectively. The properties of these copolymers were characterized by using IR spectroscopy, proton nuclear magnetic resonance （1H-NMR） spectroscopy, gel permeation chromatography （GPC）, X-ray diffraction and differential scaning calorimetry （DSC）. PLA-Hpr-PEG are amorphous copolymers. Copolymers showed increasing water uptake capacity with increasing PEG percentage in the feed, which result in an increasing degradable rate in phosphate buffer solution （pH 7.4） at 37℃.

EMS诱变菜豆子叶节定向筛选耐羟脯氨酸(HYP)突变体

以双丰一号菜豆子叶节为外植体,利用化学诱变剂EMS处理引发基因突变,进行突变体定向筛选抗逆突变细胞系。以0.17%EMS诱变处理2 h,子叶节的不再生率接近50%;以羟脯氨酸最大致死量3.5 mmol/L为筛选压力,定向筛选抗逆突变体。结果显示,在羟脯氨酸筛选培养基上,经EMS诱变的子叶节存活31个,突变率为8.61%,存活的子叶节再生率为74.19%,成功获得耐羟脯氨酸的突变株系。

绞股蓝提取液对光老化模型小鼠皮肤组织中SOD活性MDA和HYP含量影响的实验研究

目的:研究内服绞股蓝对紫外线照射引起的无毛小鼠皮肤光老化模型的影响。方法:昆明无毛小鼠随机分成5组,正常组(A)、模型对照灌胃组(B)、低剂量灌胃组(C)、中剂量灌胃组(E)、高剂量灌胃组(F)。模拟UV长期照射,造成皮肤光老化模型。给药组照射同时灌胃绞股蓝提取液。采用病理组织切片制片技术,HE染色以40倍光镜观察真皮结构改变,并以生化分析方法测定SOD活性,HYP及MDA含量。结果:绞股蓝C、D、E组对SOD活性均有提高作用(P<0.05)。D、E组可提高羟脯氨酸含量,降低MDA含量。结论:绞股蓝提取液具有拮抗紫外线照射引起的无毛小鼠皮肤光老化的作用。

活化剂HYP协同rSjc26GST的抗血吸虫性肉芽肿的效果及其机制的研究

目的探讨活化剂HYP协同rSjc26GST的抗肝内血吸虫卵肉芽肿病变的效果及机制。方法采用活化剂HYP联用rSjc26GST、福氏佐剂(FCA)联用rSjc26GST、单用HYP和单用福氏佐剂分别免疫BALB/c小鼠,末次免疫后1周,免疫和对照组小鼠均用日本血吸虫尾蚴攻击感染,7周后测定其免疫效应,观察其抗血吸虫卵肉芽肿病变的效果。结果活化剂HYP联用rSjc26GST组的总虫卵数和成熟虫卵数减少率分别为55.37%和54.31%,均显著高于FCA联用rSjc26GST组的30.3096和38.07%;该两组肝内虫卵肉芽肿的直径及面积分别下降36.86%和35.25%及62.59%和60.44%;两组IgG和IgG1水平均明显升高。结论活化剂HYP协同rSjc26GST比FCA具有更好的抗肝内血吸虫卵肉芽肿病变的效果,其机制与体液免疫(IgG和IgG1)的增强密切相关。

痹祺胶囊对实验性骨关节炎大鼠NO、HYP的影响

[目的]观察痹祺胶囊对实验性大鼠膝骨关节炎(OA)一氧化氮(NO)、羟脯氨酸(HYP)的影响。[方法]选Wistar大鼠,木瓜蛋白酶膝关节注射造模,痹祺胶囊灌胃治疗8周,测定血清NO、HYP和尿HYP的含量。[结果]模型组造模后第1天尿HYP明显升高[(17.07±4.10)mg/L],第15天[(9.92±3.09)mg/L]和第64天[(13.77±1.32)mg/L]与正常组无明显差异。模型组大鼠血清NO含量明显升高(22.64μmol/L);痹祺胶囊高、中治疗组能明显降低大鼠血清NO含量(18.24μmol/L和19.14μmol/L)。[结论]痹祺胶囊具有降低OA大鼠血清NO的含量,减轻由NO介导的对关节软骨的损伤作用。

玉屏风散对大鼠光老化皮肤HYP含量、LC数量及形态的影响

目的:探讨玉屏风散是否对大鼠光老化皮肤具有保护作用。方法:将大鼠随机分成6组,除正常对照组外,模拟UV连续照射60天,造成皮肤光老化模型,给药组照射同时分别灌胃高、中、低剂量的玉屏风散提取液,模型组灌胃等剂量生理盐水。第61天碱水解法测定皮肤组织中羟脯胺酸(HYP)含量,免疫组织化学染色法观察表皮郎格罕斯细胞(LC)数量及形态。结果:模型组HYP含量明显减少;模型+玉屏风散高、中、低剂量组HYP含量均有所提高,并呈剂量依赖(分别是P<0.01,P<0.05)。模型+玉屏风散高剂量组LC数量较多,树突状结构明显,排列较规则;模型+玉屏风散中、低剂量组对LC数量、形态及排列也有不同程度的改善作用。结论:玉屏风散对紫外线照射引起的大鼠皮肤光老化具有保护作用。其机制可能是一方面玉屏风散具有清除自由基、促进表皮细胞分裂,增加真皮胶原蛋白合成的作用;另一方面玉屏风散能够减少表皮LC的迁移,保护其树突状结构。

活化剂HYP协同“425”制剂的抗日本血吸虫虫卵肉芽肿病变免疫效应机理的研究

采用流式细胞仪测定Ｔ细胞亚群和脾脏Ｂ淋巴细胞电泳对应用活化剂ＨＹＰ协同“４２５”制剂的日本血吸虫感染的ＢＡＬＢ／ｃ小鼠进行细胞和体液免疫机能测定，并与单一用“４２５”制剂和感染对照动物进行比较。实验结果表明，ＨＹＰ协同“４２５”制剂应用后２ｗｋ，相当于感染后６ｗｋ，ＣＤ４＋／ＣＤ８＋淋巴细胞亚群的比值为６．８２，显著高于后两组的１．８７和１．７９，脾脏中Ｂ淋巴细胞总频率达７５％，高于单一用“４２５”制剂和感染对照组的６２．５％和６４．５％，特异性抗体水平（ＯＤ均值）为０．５５±０．１１，分别高于后两组的０．３５±０．０３和０．３７±０．０７。应用后３～４ｗｋ，相当于感染后７～８ｗｋ，ＣＤ４＋／ＣＤ８＋比值为１．９８～３．４７，后两组则分别为１．３２～１．５９和１．４８～１．７０，Ｂ淋巴细胞数３组均相近，但前组抗体水平（ＯＤ均值）的升高率（８２．２２％～９１．１８％）仍高于后两组。由此提示，活化剂ＨＹＰ协同“４２５”制剂诱导出的抗日本血吸虫虫卵肉芽肿的免疫效应，显著减弱血吸虫虫卵肉芽肿病变是与增强其细胞和体液免疫功能密切相关，具有明显的理论意义。

黄芪提取液对光老化无毛小鼠皮肤组织HYP MDA含量及SOD活力的影响

目的:实验旨在研究黄芪提取液是否具有抗皮肤光老化的作用及其机理。材料与方法:将昆明种无毛小鼠随机分成5组,除正常对照组外,模拟UV长期照射,造成皮肤光老化模型,给药组照射同时分别灌胃高、中、低剂量的黄芪提取液,模型组灌胃等剂量生理盐水。对其皮肤组织HYP含量、MDA含量、SOD活力进行测定,以40倍光镜观察真皮结构改变,结果:模型组无毛小鼠皮肤组织SOD活性明显降低,羟脯氨酸含量显著减少,MDA含量明显增加(P<0.05),病理切片呈现光老化状态。模型+黄芪高剂量组的皮肤羟脯氨酸及MDA含量与正常组无显著性差异。与模型组比较,模型+黄芪高剂量组对SOD活性有提高作用(P<0.05),模型+黄芪中剂量、模型+黄芪低剂量组可提高羟脯氨酸含量,降低MDA含量,并呈剂量依赖(分别是P<0.05,P<0.01)。结论:黄芪提取液对紫外线照射引起的无毛小鼠皮肤光老化具有保护作用。其机制可能是一方面黄芪本身具有清除自由基的作用;另一方面黄芪可以通过提高SOD活力来达到抗氧化的目的。

生地提取物对肺纤维化大鼠肺系数及肺组织HYP的影响

目的研究生地提取物对肺纤维化模型大鼠肺系数及肺组织羟脯氨酸(HYP)的影响。方法博莱霉素气管内注入法复制大鼠肺纤维化模型,随机分为正常组、假手术组、模型组、生地注射组、生地口服组和甲强龙组,观察大鼠肺系数和肺组织HYP指标。结果与模型组比较,生地注射组和生地口服组第28天肺系数降低(P<0.05,P<0.001),各给药组第3天HYP均有降低趋势,生地注射组和生地口服组第28天HYP降低(P<0.05,P<0.01)。结论生地提取物可改善肺纤维化大鼠肺系数,从而有效治疗肺纤维化,其机制可能和抑制HYP胶原蛋白的表达有关。

本草养颜膏对老龄大鼠皮肤SOD和Hyp的影响

目的:研究本草养颜膏对老龄大鼠皮肤组织中超氧化物歧化酶(SOD)活性及羟脯氨酸(Hyp)含量的影响。方法:将SPF级SD雌性老龄大鼠50只随机分为5组:阳性对照组、模型对照组和本草养颜膏治疗高、中、低剂量组,每组10只;另取SPF级SD雌性青龄大鼠10只,作为空白对照组。分别给予维生素E、本草养颜膏基质及高、中、低剂量的本草养颜膏,于第2疗程(20天)和第3疗程(30天)结束时以生化分析法测定皮肤组织中SOD活性和Hyp含量。结果:至第3疗程结束时,本草养颜膏高、中、低剂量组均能明显提高老龄大鼠皮肤组织中SOD活性(P<0.01)和Hyp含量(P<0.01),尤其以中剂量组为最佳。结论:本草养颜膏对大鼠皮肤衰老具有延缓的作用。其机制可能是通过提高皮肤中SOD活性及Hyp含量,增强机体抗氧化和清除自由基以及合成胶原蛋白的能力,来达到延缓皮肤衰老的作用。

活化剂HYP协同“425”制剂抗日本血吸虫卵肉芽肿病变的效果

目的:了解活化剂HYP能否增强“425”制剂抗日本血吸虫卵肉芽肿病变的效果。方法:采用皮肤感染日本血吸虫尾蚴制成BALB/c肝内虫卵肉芽肿病变模型,通过检测和计算肝内肉芽肿平均直径、面积和体积观察“425"制剂的治疗作用,以及活化剂HYP的协同增效作用。结果:用药2周与对照组相比,其肝内虫卵肉芽肿平均直径下降率42.12％,平均面积下降65.71％,平均体积下降80.4％,用药3～4周时平均直径、面积和体积下降率分别为31.61％～47.48％,55.14％～73.19％和71.15％～75.86％。单用“425”制剂2～3周,肝内肉芽肿大小与感染对照鼠无显著性差异,应用4周时其平均面积和体积分别下降14.15％和7.76％。结论:活化剂HYP具有明显的协同增强抗宿主肝内日本血吸虫卵肉芽肿病变的效果。

重组大肠杆菌BL21(pUC19-Hyp)产羟脯氨酸的补料分批培养

利用自主构建的组成型重组大肠杆菌BL21(pUC19-Hyp)为出发菌株,运用间歇流加、指数流加和恒速流加3种流加C源的方式进行补料分批培养。结果表明:在装液量为4 L的7 L发酵罐中,以0.30 g/min恒速流加为最优,在发酵44 h时,羟脯氨酸的质量浓度达到最高,为42.50 g/L,脯氨酸转化率为81%,此时细胞干质量为21.33g/L,残糖质量浓度为0.17 g/L。L-羟脯氨酸含量与摇瓶发酵时的1.39 g/L相比,提高了大约30倍,比日本株式会社的发酵产量提高了1.50 g/L,发酵过程中糖酸转化率约为4.0∶1。发酵液中的氨基酸分析结果表明,除脯氨酸、羟脯氨酸外的其他氨基酸质量浓度均低于0.1 g/L,发酵液中主要氨基酸为脯氨酸和羟脯氨酸。