甘肃鼢鼠骨骼19种氨基酸含量的测定

用121型氨基酸分析仪对甘肃鼢鼠(Myospalax cansus)躯干、四肢、头骨骨骼中19种氨基酸含量进行测定,并与高原鼢鼠及其他哺乳动物骨骼中氨基酸含量进行差异显著性分析。结果表明,甘肃鼢鼠各部位骨骼中19种氨基酸含量均为甘氨酸最高,谷氨酸次之,胱氨酸最低。同一氨基酸在甘肃鼢鼠不同骨骼部位含量顺序为躯干>四肢(头骨。甘肃鼢鼠骨骼总氨基酸含量低于塞隆骨,与虎骨相近,高于其他哺乳动物。其中羟脯氨酸含量明显高于虎骨、塞隆骨等。     

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。     

L-羟脯氨酸用于小鼠骨折愈合的实验研究

让骨折的小白鼠饮用L 羟脯氨酸水溶液 ,然后通过X线拍片观察其骨折的愈合情况 ,并与对照组进行比较 ,结果表明 :L 羟脯氨酸有促进骨折愈合的作用 ,而且安全、无毒。     

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。     

神经酰胺合酶-神经酰胺通路在青蒿琥酯抑制肝纤维化中的作用

目的: 探讨神经酰胺通路在青蒿琥酯(Art)抑制肝纤维化过程中的作用。方法: 将肝星状细胞(LX-2)分为细胞对照组、Art 350 μmol/L组、神经酰胺合酶阻断剂(FB1) 6 μmol/L组及FB1 6 μmol/L + Art 350 μmol/L合用组。每组7个复孔,作用24 h后,收集细胞及上清液进行检测。Western blot法测定神经酰胺合酶2(LASS2)、过氧化物酶体增殖物激活受体-γ(PPAR-γ)、Caspase-3蛋白的表达, HPLC-FLD法对神经酰胺(Cer)的含量进行测定,MTT法检测LX-2的增殖情况,酶消化法检测羟脯氨酸(Hyp)的含量。结果: 与细胞对照组比较,Art处理组神经酰胺合酶的蛋白表达、神经酰胺含量显著增加、LX-2细胞的增殖明显抑制、PPAR-γ和Caspase-3蛋白表达上调、羟脯氨酸分泌抑制 (P＜0.05);FB1处理组神经酰胺合酶的蛋白表达和神经酰胺含量显著降低、LX-2细胞增殖显著增加、PPAR-γ和Caspase-3蛋白表达下调、羟脯氨酸分泌增加(P＜0.05);与Art单用组比较,两药合用可显著降低Art对神经酰胺合酶蛋白的表达和神经酰胺含量升高的作用,减弱Art对LX-2细胞增殖、PPAR-γ、Caspase-3蛋白表达及羟脯氨酸分泌的作用(P＜0.05)。结论: 青蒿琥酯可通过神经酰胺合酶-神经酰胺通路增加细胞中神经酰胺水平,发挥抑制肝纤维化的作用。     

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     

呲效隆对花生光合作用及产量的影响

花生结荚期施用0.1～10 mg/L吡效隆(4PU-30)溶液,能增加叶片的厚度,提高叶片叶绿素含量和光合速率,促使叶片中的同化物向荚果运输和积累增多,从而促进了荚果生长和发育,使结荚率、饱果率以及百果重和百仁重增加,最终使单株荚果产量增产12.6％左右,吡效隆的最适浓度为 1 mg/L。     

L-羟脯氨酸用于减肥的实验研究

用添加L -羟脯氨酸的饲料对营养性肥胖Wistar大鼠和昆明种小鼠进行实验 ,与对照组比较体重、体脂、血脂和摄食量的变化。结果表明L -羟脯氨酸能减少营养性肥胖大鼠的体重和体脂 ,降低血脂 ,且无毒 ,不影响食欲 ,在减肥方面有良好的发展前景。     

二维制备高效液相色谱分离塞隆骨强极性多肽

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。     

脯氨酸-4-羟化酶在大肠杆菌中的密码子优化表达及对反式-4-羟脯氨酸生物合成的作用

为了使脯氨酸-4-羟化酶基因在重组大肠杆菌中得到高表达,通过调整大肠杆菌密码子偏好性以及mRNA二级结构,使得脯氨酸-4-羟化酶基因得到优化。将优化后的脯氨酸-4-羟化酶基因插入含有色氨酸串联启动子的p UC19质粒,构建重组质粒p UC19-ptrp2-Hyp,并导入大肠杆菌BL21(DE3)中。在摇瓶水平,重组菌以L-脯氨酸为底物发酵8 h,可积累(0.492±0.034)g/L的反式-4-羟脯氨酸。在发酵罐水平,通过补料分批发酵来提高反式-4-羟脯氨酸的产量,当补糖速率为18 g/h时,反式-4-羟脯氨酸的产量高达42.5 g/L,反式-4-羟脯氨酸产率为0.966 g/(L·h)。     

Liquid chromatographic determination of hydroxyproline in tissue samples

We describe a reversed-phase assay of hydroxyproline in rat lung tissue using sarcosine for the internal standard and pre-injection reaction with both o-phthalaldehyde (OPA) and 9-fluorenylmethylchloroformate (FMOC). Intra-assay variability in the concentration range of 25-500 microM hydroxyproline was less than 1%. Normal rat (left) lung was found to have a hydroxyproline content of 1.08+/-0.18 mg/lung. This ability to measure minute amounts of hydroxyproline is being applied to the measure of collagen and pathological fibrosis.     

Mechanical properties of human tracheal cartilage.

Biomechanical changes in airway cartilage could influence the mechanics of maximal expiratory flow and cough and the degree of shortening of activated airway smooth muscle. We examined the tensile stiffness of small samples of human tracheal cartilage rings in specimens obtained at autopsy from 10 individuals who ranged in age from 17 to 81 yr. The tensile properties of the cartilage were compared with its content of water (%water), glycosaminoglycans (chondroitin sulfate equivalents, mg/mg dry wt), and hydroxyproline content (mg hydroxyproline/mg dry weight). The average values for tensile stiffness ranged between 1 and 15 MPa and increased significantly with increasing age [tensile stiffness = 0.19 x (age in yr) + 2.02; r = 0.83, P less than 0.05]. The outermost layer of cartilage was the most stiff in all individuals, and the deeper layers were progressively less stiff. Water content and hydroxyproline content both decreased with increasing age. Thus tensile stiffness correlated inversely with water content and hydroxyproline content [tensile stiffness = -0.83 x (%water) + 16.4; r = 0.82, P less than .05 and tensile stiffness = -342 x (hydroxyproline content) + 25; r = 0.87, P less than 0.05]. Total tissue content of glycosaminoglycans did not change with age, although changes in glycosaminoglycan type and proteoglycan structure with increasing age have been described. We conclude that there are age-related changes in the biomechanical properties and biochemical composition of airway cartilage that could influence airway dynamics.     

Free and small peptide-bound [14C]hydroxyproline synthesis in rat liver in vitro in CCl4-induced hepatic fibrosis

To clarify the process of free and small peptide-bound hydroxyproline synthesis in hepatic fibrogenesis, we measured the in vitro synthesis of [14C]hydroxyproline in the 67% ethanol soluble fraction in rat liver slices, together with hepatic protein-bound [14C]hydroxyproline synthesis. In control rat liver, the amount of free and small peptide-bound [14C]hydroxyproline synthesized was 13.1 +/- 2.6 10(-4) x dpm/g liver/3 hr. In the CCl4-treated rat liver, where the hepatic hydroxyproline content was increased 4.6-fold, the protein-bound [14C]hydroxyproline synthesis was significantly increased 1.5-fold, but free and small peptide-bound [14C]hydroxyproline synthesis was decreased into 70%. There was a significant inverse correlation between free and small peptide-bound [14C]hydroxyproline synthesis, and hepatic hydroxyproline content. These results suggest that the combination of an increase in collagen synthesis and a decrease in free and small peptide-bound [14C]hydroxyproline synthesis contributes to rapid accumulation of collagen in hepatic fibrosis.     

解毒活血汤促进肛周脓肿术后创面愈合的效果分析

目的:研究解毒活血汤治疗肛周脓肿术后患者创面愈合的临床效果及可能机制。方法:选择2017年1月～2019年1月我院肛肠科三病区收治的300例肛周脓肿患者,根据其就诊的顺序,随机数字表法均分为两组,各150例,两组均采取一期肛周脓肿根治术治疗,对照组于术后采用高锰酸钾溶液坐浴疗法,观察组于术后联合服用解毒活血汤治疗。均治疗4 w后,比较两组创面愈合的有效率,肛周脓肿患者创面的愈合时间和视觉模拟评分(visual analogue scale,VAS)评分,治疗前后肉芽组织中的血红蛋白含量、羟脯氨酸含量、血管内皮生长因子水平以及组织微血管计数。结果:治疗后,观察组创面愈合的总有效率明显高于对照组(90.67 vs. 71.33%,P0.05);观察组肛周脓肿患者创面愈合的时间明显短于对照组(P0.05)。对照组治疗前后的肉芽组织中的血红蛋白含量、羟脯氨酸含量、血管内皮生长因子水平以及组织微血管计数无明显的差异(P0.05),观察组治疗后的肉芽组织中的血红蛋白含量、羟脯氨酸含量、血管内皮生长因子水平以及组织微血管计数明显高于治疗前和对照组(P0.05)。观察组术后3 d、1 w和2 w的VAS评分均明显低于对照组(P0.05)。结论:解毒活血汤可能通过提升肉芽组织中的血红蛋白含量、羟脯氨酸含量、血管内皮生长因子水平以及组织微血管计数有效促进肛周脓肿术后的创面愈合,减轻术后疼痛。     

Effects of perturbed geomagnetic field on connective tissue metabolism and some systems of homeostasis after hydroxyapatite implantation

The influence of perturbed geomagnetic field (PGMF) on glucocorticoid, epinephrine, hydroxyproline content in intact rats and after intramuscular implantation of demineralized bone matrix and combined (hydroxyapatite/bone meal) samples was investigated. It was found that the exposure of the intact rats to PGMF led to decrease in free and peptide-bound hydroxyproline in plasma. In the rats with hydroxyapatite/bone meal samples placed in ectopic sites the exposure to PGMF normalized hydroxyproline, glucocorticoid and epinephrine content.     

Effects of food restriction on stretch induced muscle hypertrophy in chickens of various ages

1. The patagialis muscle of three groups of different aged chickens was subjected to passive stretch by placing an inflexible sleeve over the left wing, and the right wing served as the control. 2. Food was removed from the animals 2 days prior to applying passive stretch. After 6 days of passive stretch on young 7-week-old animals, the stretched muscle was 200% larger than the control. 3. During the restriction, control muscle lost 69% of its weight and the stretched muscle maintained its original weight. 4. Changes in muscle DNA and hydroxyproline content were similar to changes in muscle weight but hydroxyproline content lagged changes in muscle weight. 5. Restricted 10-month-old chickens responded to passive stretch with an absolute increase in muscle mass which was accompanied by increases in protein, DNA and hydroxyproline content. 6. Muscle from the 28-month-old chickens did not respond to either 11 days of restriction or passive stretch. 7. The results of the present study indicate that as the chicken grows older, the ability of the muscle to respond to stimuli, stretch and/or starvation, is reduced.     

Influence of bleomycin on the metabolism of dermal collagen in albino rats

The metabolism of collagen in male rats by treatment with bleomycin was studied following the injection of [3H]proline and the determination of specific and total activity of [3H]hydroxyproline in skin collagen fractions and urine. In the case of the bleomycin-treated animals, there was found to be an increase in the neutral salt soluble collagen content with no change in insoluble collagen content as compared to the control group. The specific and total radioactivity of [3H]hydroxyproline in soluble and insoluble collagen fractions was also increased. Examination of [3H]hydroxyproline activity in soluble and insoluble collagen showed that the conversion of soluble to insoluble collagen was improved by the bleomycin-treated group. It was found that this was accompanied by a decrease in urinary excretion of total hydroxyproline and [3H]hydroxyproline during the first 12 hr after the administration of [3H]proline. Therefore, the results of the present investigation clearly indicate that the maturation of soluble to insoluble collagen is promoted and accompanied by a decrease in the catabolism of soluble collagen in the bleomycin-treated animals. In addition, administration of bleomycin increased the synthesis of collagen.     

Subunit structure of wheat germ agglutinin

Cells isolated by enzymic digestion of embryonic tendon were incubated under N₂ so that they synthesized and accumulated the unhydroxylated form of procollagen which is known as protocollagen and which is largely comprised of pro-α chains linked by interchain disulfide bonds. The cells were then exposed to O₂ so that the intracellular protocollagen was hydroxylated and secreted as procollagen. When the hydroxylation was allowed to proceed at 31° or 34°, the procollagen secreted into the medium was triple-helical but its hydroxyproline content was less than two-thirds and its hydroxylysine content was less than half the control. Even when the hydroxylation was allowed to occur at 37°, the procollagen secreted by the cells was under-hydroxylated by about 15% in terms of its hydroxyproline content and about 45% in terms of its hydroxylysine content. The results may have consequences for collagen synthesis by tendons and similar tissues $\text{in vivo}$, since temporary anoxia in such tissues may well lead to the synthesis of a less stable procollagen or to fibers of decreased tensile strength.     

HRGP在小麦抗寒锻炼过程中的变化及其与抗寒性的关系

强抗寒小麦品种(R-025、中品94-19、品83-1、品83-2、品83-3、米罗诺夫808)经抗寒锻炼后,其幼苗体内的游离脯氨酸、细胞壁结合的羟脯氨酸和糖蛋白含量发生了明显的变化.游离脯氨酸含量比未经抗寒锻炼处理时增加5～32倍,细胞壁结合的羟脯氨酸含量比对照增加1.77～2.17倍,糖蛋白含量比对照增加4.68～9.72倍,而不抗寒小麦品种(中国春、冬103)增加量较小.脯氨酸积累进程各个品种间差异比较大,品83-1、品83-2积累较快,抗寒锻炼第21d时达到最高峰,而R-025在第56d达到最高峰.脯氨酸含量与小麦品种抗寒性相关不显著(相关系数为0.3462),而羟脯氨酸含量、糖蛋白含量与小麦品种抗寒性相关显著,相关系数分别为0.6491和0.7039.从小麦细胞壁纯化得到了2种伸展蛋白Extensm1和Ex-tensin2,其含量都和小麦品种抗寒性呈正相关.Extensin1是分子量为28kD、羟脯氨酸为主要成份(32mo1%)的富含羟脯氨酸糖蛋白.     

Effects of 900 MHz Radiofrequency Radiation on Skin Hydroxyproline Contents

The present study aimed to investigate the possible effect of pulse-modulated radiofrequency radiation (RFR) on rat skin hydroxyproline content, since skin is the first target of external electromagnetic fields. Skin hydroxyproline content was measured using liquid chromatography mass spectrometer method. Two months old male wistar rats were exposed to a 900 MHz pulse-modulated RFR at an average whole body specific absorption rate (SAR) of 1.35 W/kg for 20 min/day for 3 weeks. The radiofrequency (RF) signals were pulse modulated by rectangular pulses with a repetition frequency of 217 Hz and a duty cycle of 1:8 (pulse width 0.576 ms). A skin biopsy was taken at the upper part of the abdominal costa after the exposure. The data indicated that whole body exposure to a pulse-modulated RF radiation that is similar to that emitted by the global system for mobile communications (GSM) mobile phones caused a statistically significant increase in the skin hydroxyproline level (p = 0.049, Mann–Whitney U test). Under our experimental conditions, at a SAR less than the International Commission on Non-Ionizing Radiation Protection safety limit recommendation, there was evidence that GSM signals could alter hydroxyproline concentration in the rat skin.