不同月龄大鼠椎间盘退变与多效生长因子表达的关系

目的观察不同月龄大鼠椎间盘的形态学变化并检测椎间盘中多效生长因子（pleiotrophin,PTN）的表达,探讨PTN与椎间盘退变的关系。方法取Wistar大鼠50只,以1,3,6,12,18个月龄不同分为5组,每组10只。采用苏木精-伊红染色观察椎间盘的形态学变化。采用SABC免疫组织化学方法,检测椎间盘中PTN的表达情况;结果（1）随着月龄的增加,椎间盘组织结构紊乱的程度逐渐增加,髓核内基质降解、正中出现空腔,胶原纤维增生、粗大、排列紊乱、并可见纤维断裂或缺失。（2）随着大鼠月龄的增加（1-12月龄）,椎间盘细胞中PTN的表达有逐渐减低的趋势,但至18月龄,PTN表达又有所增加;6和12月龄组椎间盘细胞中PTN的表达显著低于1月龄组,而18月龄组PTN的表达显著高于12月龄组。同月龄组椎间盘细胞中,PTN在终板的表达高于髓核和纤维环,髓核和纤维环中PTN的表达未见明显差异。结论大鼠椎间盘结构随月龄增加发生退行性变,PTN参与了大鼠椎间盘的退变,并可能通过促进椎间盘组织中新生血管的形成,延缓椎间盘的退变。     

Sox9治疗兔椎间盘退变的效果及机制研究

目的:探究Sox9用于治疗椎间盘退变的效果及调控机制。方法:将Ad-sox9和Ad-GFP各20μL分别转染至椎间盘退变兔的髓核组织中,转染后3、7、30、60天取材,采用免疫组化、免疫荧光和MRI等研究方法检测椎间盘髓核组织中II型胶原、蛋白多糖的表达情况,并分析对椎间盘退变的改善情况。结果:免疫组化染色显示sox9组中椎间盘髓核组织中II型胶原、蛋白多糖的表达明显升高,MRI显示sox9组椎间盘T2像信号有明显改善(P<0.05)。结论:体内转染腺病毒介导的sox9基因能够增加椎间盘内II型胶原和蛋白多糖的表达,并抑制椎间盘的退变进程。     

Peroxiredoxin Ⅱ在退变椎间盘髓核中的表达及其临床意义

目的:检测PeroxiredoxinⅡ在腰椎间盘髓核组织中的表达,分析其在椎间盘退变中的临床意义。方法:用蛋白免疫印迹(Western blot)的方法检测PeroxiredoxinⅡ在正常、突出及脱出腰椎间盘髓核中的表达情况。结果:PeroxiredoxinⅡ在退变椎间盘髓核中表达丰富,而在正常椎间盘髓核中表达微弱,两者比较差异显著(P<0.05),在突出和脱出椎间盘髓核中表达无显著性差异(P>0.05)。结论:PeroxiredoxinⅡ在正常及退变腰椎间盘髓核组织中差异表达。     

PeroxiredoxinⅡ在退变椎间盘髓核中的表达及其临床意义

目的：检测PeroxiredoxinⅡ在腰椎间盘髓核组织中的表达,分析其在椎间盘退变中的临床意义。方法：用蛋白免疫印迹（Western blot）的方法检测PeroxiredoxinⅡ在正常、突出及脱出腰椎间盘髓核中的表达情况。结果：PeroxiredoxinⅡ在退变椎间盘髓核中表达丰富,而在正常椎间盘髓核中表达微弱,两者比较差异显著（P〈0.05）,在突出和脱出椎间盘髓核中表达无显著性差异（P〉0.05）。结论：PeroxiredoxinⅡ在正常及退变腰椎间盘髓核组织中差异表达。     

Peroxiredoxin Ⅱ在退变椎间盘髓核中的表达及临床意义

目的:检测Pcroximdoxin Ⅱ在腰椎间盘髓核组织中的表达,分析其在椎间盘退变中的临床意义.方法:用蛋白免疫印迹(Western blot)的方法检测Peroxiredoxin Ⅱ在正常、突出及脱出腰椎间盘髓核中的表达情况.结果:Peroxiredoxin Ⅱ在退变椎间盘髓核中表达丰富,而在正常椎问盘髓核中表达微弱,两者比较差异显著(P<0.05),在突出和脱出椎间盘髓核中表达无显著性差异(P>0.05).结论:Peroxiredoxin Ⅱ在正常及退变腰椎间盘髓核组织中差异表达.     

缺氧诱导因子-1α和血管内皮生长因子在突出腰椎间盘中的表达及意义

目的探讨缺氧诱导因子-1α（hypoxia—induciblefactor-1α,HIF-1α）和血管内皮生长因子（vascular endothelial growth factor,VEGF）在突出腰椎间盘组织中的表达及意义。方法采用链霉亲和素-过氧化物酶复合物（SABC）免疫组化方法,测定40例腰椎间盘突出症患者椎间盘组织中HIF-1α和VEGF的表达情况。结果退变椎间盘组织中HIF-1α和VEGF呈高表达,HIF-1α和VEGF在髓核的表达显著高于纤维环;纤维环破裂型显著高于纤维环完整型;各组中HIF-1α和VEGF的表达均高度相关。结论HIF-1α和VEGF共同参与了椎间盘退变;HIF-1α可能通过上调VEGF的表达来促进椎间盘组织中新生血管的形成,进而延缓椎间盘退变的发生。     

Bcl2、Fas在人颈椎间盘软骨终板、纤维环及髓核中的表达及意义

目的 检测正常和突出的人颈椎间盘软骨终板、纤维环及髓核中Bcl2、Fas的表达情况,探讨其在各组中不同表达的意义。方法 先将各正常及突出颈椎间盘在解剖显微镜下分出软骨终板、纤维环、髓核并进行苏木精-伊红（HE）染色行形态学观察,再用免疫组化（SABC法）检测Bcl2、Fas在各组中的表达情况,并进行计算机显像系统图像分析。结果 Bcl2的表达在正常颈椎间盘的软骨终板、纤维环、髓核中均高于突出颈椎间盘的各相应组;Fas的表达在正常颈椎间盘的软骨终板、纤维环、髓核中均低于突出颈椎间盘的各相应组;在异常颈椎间盘的软骨终板中Bcl2的表达低于其在纤维环和髓核中的表达（P〈0.01）;Fas的表达高于其在纤维环和髓核中的表达（P〈0.01）;Bcl2及Fas的表达在纤维环和髓核中的差别无统计学差异（P〉0.05）。结论 细胞凋亡是椎间盘退变的重要原因,且凋亡可能始于软骨终板。     

椎间盘退变纤维环及髓核异常表达基因的比较及生物信息学分析

目的:通过对已公开发表的基因芯片表达谱数据进行研究,探究椎间盘退变过程中纤维环与髓核组织的基因表达差异,并采用生物信息学方法对差异进行分析。方法:经GEO数据库选取两组椎间盘退变相关的基因芯片表达谱数据GSE23130及GSE67567,GSE23130所研究标本来源于正常及退变纤维环组织,GSE67567标本来源于正常及退变髓核组织。对上述数据系列进行质量分析,GSE23130及GSE67567各有10例样本数据被纳入实验。采用Gene Spring 13.0软件对GSE23130正常及退变纤维环间差异表达基因及GSE67567正常及退变髓核间差异表达基因分别进行筛选,利用KEGG PATHWAY和DAVID功能注释簇集分析分别对GSE23130及GSE67567上调及下调基因进行生物信息学分析。结果:GSE23130及GSE67567各筛选出差异表达基因3182个和3017个,其中135个基因在上述两个基因表达谱数据中均存在差异表达。针对两组数据进行的KEGG PATHWAY分析发现TGF-beta signaling pathway和regulation of apoptosis等数个相同的生物学通路及DAVID功能注释簇集;此外,还发现了数个与GSE23130及GSE67567单独相关的DAVID功能注释簇集。结论:椎间盘退变过程中纤维环及髓核组织内基因表达情况存在差异,两种组织内发生的生物过程不尽相同。某些生物学过程在两种组织内均出现异常改变,这些生物学过程中的异常变化可能是椎间盘退变的关键环节,值得进行深入研究。     

退变腰椎间盘中IL-17表达的变化及其意义

目的:研究退变的椎间盘组织中IL-17表达的变化及其与椎间盘退变严重程度之间的关系。方法:收集退变椎间盘标本23例,正常椎间盘标本12例,通过免疫组织化学染色、免疫荧光染色、实时-定量PCR(RT-PCR)和酶联免疫吸附实验(ELISA)从细胞、蛋白和基因水平检测椎间盘组织中IL-17和孤独受体(retinoid-related orphan receptor,RORγt)的表达。结果:免疫组化染色显示退变椎间盘组织中IL-17阳性细胞比例较对照组明显增高,有统计学差异(P<0.05);免疫荧光染色显示退变椎间盘组织中Th17细胞含量明显增多(P<0.05);退变的椎间盘组织中IL-17和RORγt m RNA的相对表达量较对照组增加,有统计学差异(P<0.001),且两者之间呈显著正相关(r=0.6919,P<0.001);退变的椎间盘组织中IL-17的含量较对照组明显增加(P<0.01),且与椎间盘退变的严重程度呈正显著相关(r=0.4714,P<0.01)。结论:IL-17含量增加参与了腰椎间盘退变的病理过程,并且可能对椎间盘退变起促进作用。     

Cathepsin L与MMP-3在兔椎间盘退变中相关性研究

目的:探讨组织蛋白醇L(Cathepsin L)与基质金属蛋白酶-3(MMP-3)在兔椎间盘退变中的相关性.方法:健康大白兔60只随机分为实验组、阴性对照组、空白对照组,实验组分为三个亚组,椎间盘内注射3种不同浓度的Cathepsin L,阴性对照组中在椎间盘内注射生理盐水,空白对照组不做任何处理.采用HE粢色组织学观察椎间盘的退变特征,免疫组织化学S-P法检测椎间盘组织中MMP-3的表达.结果:HE染色组织学观察6周时在实验组椎间盘中,组织学观察到不同程度的椎间盘退变特征:□核脊索细胞逐渐减少,并逐渐被纤维软骨组织替代,纤维环与髓核之间失去明显的界限.免疫组织化学染色结果,3周、6周时实验组MMP-3的阳性率均高于空白对照组及阴性对照组(P＜0.05).结论:Cathepsin L与MMP-3共同参与了兔椎间盘的退变,Cathepsin L可能促进MMP-3在椎间盘内的生成,从而导致椎间盘的退变.     

Glycolipid Degradation in Leaves of the Thermophilic Cucumis sativus as Affected by Light and Low-Temperature Treatment

Treatment of Cucumis leaf discs with light and low temperature (1°C) resulted in degradation of the total lipids. In addition, a decrease of the linolenic acid content of the glycolipids of leaves and leaf discs took place while at the same time an increase in the content of unidentified degradation products from the glycolipids was observed. The decrease of the linolenic acid content was not due to galactolipase activity, since no monogalactosyl diglyceride acyl hydrolase activity was found. Dark and low temperature did not alter the fatty acid composition. Blue light and low temperature resulted in a decrease of the linolenic acid content, while yellow-, red- and far red light in combination with low temperature were ineffective.     

Is the Transport of a Gadolinium-Based Contrast Agent Decreased in a Degenerated or Aged Disc? A Post Contrast MRI Study

A post contrast magnetic resonance imaging study has been performed in a wide population of low back pain patients to investigate which radiological and phenotypic characteristics influence the penetration of the contrast agent in lumbar discs in vivo. 37 patients affected by different pathologies (disc herniation, spondylolisthesis, foraminal stenosis, central canal stenosis) were enrolled in the study. The selected population included 26 male and 11 female subjects, with a mean age of 42.4±9.3 years (range 18–60). Magnetic resonance images of the lumbar spine were obtained with a 1.5 T scanner (Avanto, Siemens, Erlangen, Germany) with a phased-array back coil. A paramagnetic non–ionic contrast agent was injected with a dose of 0.4 ml/kg. T1-weighted magnetic resonance images were subsequently acquired at 5 time points, 5 and 10 minutes, 2, 4 and 6 hours after injection. Endplates presented clear enhancement already 5 minutes after injection, and showed an increase in the next 2 hours followed by a decrease. At 5 and 10 minutes, virtually no contrast medium was present inside the intervertebral disc; afterwards, enhancement significantly increased. Highly degenerated discs showed higher enhancement in comparison with low and medium degenerated discs. Discs classified as Pfirrmann 5 showed a statistically significant higher enhancement than Pfirrmann 1, 2 and 3 at all time points but the first one, possibly due to vascularization. Disc height collapse and Modic changes significantly increased enhancement. Presence of endplate defects did not show any significant influence on post contrast enhancement, but the lack of a clear classification of endplate defects as seen on magnetic resonance scans may be shadowing some effects. In conclusion, disc height, high level of degeneration and presence of Modic changes are factors which increase post contrast enhancement in the intervertebral disc. The effect of age could not be demonstrated.     

Autoinhibition of Ethylene Production in Citrus Peel Discs : SUPPRESSION OF 1-AMINOCYCLOPROPANE-1-CARBOXYLIC ACID SYNTHESIS

Wound ethylene formation induced in flavede tissue of citrus fruit (Citrus paradisi MacFad. cv. Ruby Red) by slicing was almost completely inhibited by exogenous ethylene. The inhibition lasted for at least 6 hours after removal of exogenous ethylene and was then gradually relieved. The extent of inhibition was dependent upon the concentration of ethylene (1 to 10 microliters/liter) and the duration of treatment. The increase in wound ethylene production in control discs was paralleled by an increase in 1-aminocyclopropane-1-carboxylic acid (AAC) content, whereas in ethylene-treated discs there was little increase in ACC content. Application of ACC completely restored ethylene production in ethylene-pretreated discs, indicating that the conversion of ACC to ethylene is not impaired by the presence of ethylene. Thus, autoinhibition of ethylene synthesis was exerted by reducing the availability of ACC. Ethylene treatment resulted in a decrease in extractable ACC synthase activity, but this decrease was too small to account for the marked inhibition of ACC formation. The data indicate that autoinhibition of ethylene production in citrus flavede discs results from suppression of ACC formation through repression of the synthesis of ACC synthase and inhibition of its activity.     

Physiological Loading Can Restore the Proteoglycan Content in a Model of Early IVD Degeneration

A hallmark of early IVD degeneration is a decrease in proteoglycan content. Progression will eventually lead to matrix degradation, a decrease in weight bearing capacity and loss of disc height. In the final stages of IVD degradation, fissures appear in the annular ring allowing extrusion of the NP. It is crucial to understand the interplay between mechanobiology, disc composition and metabolism to be able to provide exercise recommendations to patients with early signs of disc degeneration. This study evaluates the effect of physiological loading compared to no loading on matrix homeostasis in bovine discs with induced degeneration. Bovine discs with trypsin-induced degeneration were cultured for 14 days in a bioreactor under dynamic loading with maintained metabolic activity. Chondroadherin abundance and structure was used to confirm that a functional matrix was preserved in the chosen loading environment. No change was observed in chondroadherin integrity and a non-significant increase in abundance was detected in trypsin-treated loaded discs compared to unloaded discs. The proteoglycan concentration in loaded trypsin-treated discs was significantly higher than in unloaded disc and the newly synthesised proteoglycans were of the same size range as those found in control samples. The proteoglycan showed an even distribution throughout the NP region, similar to that of control discs. Significantly more newly synthesised type II collagen was detected in trypsin-treated loaded discs compared to unloaded discs, demonstrating that physiological load not only stimulates aggrecan production, but also that of type II collagen. Taken together, this study shows that dynamic physiological load has the ability to repair the extracellular matrix depletion typical of early disc degeneration.     

ca2 +对苹果果实细胞膜透性 保护酶活性和保护物质含量的影响

CaCl2处理“金冠”苹果果肉圆片,明显地增加Ca2+含量,降低K+外渗。不同浓度Ca2+程度不同地提高圆片超氧化物歧化酶、过氧化氢酶活性和谷胱甘肽、抗坏血酸含量,降低H2O2含量和相对电导率。本文认为：Ca2+能提高保护酶活性和保护物质含量,有利于保护细胞膜结构。     

Deferral of leaf senescence with calcium

In view of the possibility that senescence may be a consequence of the deterioration of membrane compartments in the cells of leaves, calcium was studied as a possible agent which might defer senescence. The senescence of corn leaf discs was deferred by added calcium, and the effect was additive to the cytokinin deferral of senescence. Likewise, the senescence of Rumex leaf discs was deferred by added calcium, and the effect was additive to the gibberellin deferral of senescence. Detailed experiments with corn leaf discs established that the increase in apparent free space associated with senescence was completely prevented by calcium. An increase in hydraulic permeability during senescence was likewise demonstrated, and this increase was deferred by calcium; calcium plus benzyladenine was even more effective. Each of the measured functions of leaf senescence (chlorophyll content, protein decrease, apparent free space increase, and hydraulic permeability increase) was suppressed by calcium, and the interpretation is offered that the effects are a consequence of the calcium function in maintaining cellular membranes.     

Increase of nerve growth factor levels in the human herniated intervertebral disc: can annular rupture trigger discogenic back pain?

Introduction

Nerve growth factor (NGF) has an important role in the generation of discogenic pain. We hypothesized that annular rupture is a trigger for discogenic pain through the action of NGF. In this study, the protein levels of NGF in discs from patients with disc herniation were examined and compared with those from discs of patients with other lumbar degenerative disc diseases.

Methods

Patients (n = 55) with lumbar degenerative disc disease treated by surgery were included. Nucleus pulposus tissue (or herniated disc tissue) was surgically removed and homogenized; protein levels were quantified using an enzyme-linked immunosorbent assay (ELISA) for NGF. Levels of NGF in the discs were compared between 1) patients with herniated discs (herniated group) and those with other lumbar degenerative disc diseases (non-herniated group), and 2) low-grade and high-grade degenerated discs. Patient’s symptoms were assessed using a visual analog scale (VAS) and the Oswestry disability index (ODI); the influence of NGF levels on pre- and post-operative symptoms was examined.

Results

Mean levels of NGF in discs of patients were significantly higher in herniated discs (83.4 pg/mg total protein) than those in non-herniated discs (68.4 pg/mg).No significant differences in levels of NGF were found between low-grade and high-grade degenerated discs. Multivariate analysis, adjusted for age and sex, also showed significant correlation between the presence of disc herniation and NGF levels, though no significant correlation was found between disc degeneration and NGF levels. In both herniated and non-herniated groups, pre-operative symptoms were not related to NGF levels. In the herniated group, post-operative lower extremity pain and low back pain (LBP) in motion were greater in patients with low levels of NGF; no significant differences were found in the non-herniated group.

Conclusions

This study reports that NGF increased in herniated discs, and may play an important role in the generation of discogenic pain. Analysis of patient symptoms revealed that pre-operative NGF levels were related to post-operative residual lower extremity pain and LBP in motion. The results suggest that NGF in the disc is related to pain generation, however, the impact of NGF on generation of LBP varies in individual patients.

Electronic supplementary material

The online version of this article (doi:10.1186/ar4674) contains supplementary material, which is available to authorized users.     

高温胁迫对桃和抽细胞膜透性和光合色素的影响

研究了桃和袖离休叶片在高温胁迫下细胞膜透性（电解质渗漏和Pi渗漏）和光合色素的变化及其与抗热性的关系。结果表明。①高温处理下,无论是黑暗条件下浸水24h,还是光照条件下浸水24h,两种果树的电解质和Pi渗漏都显著增加,光照有利于膜的修复;②高温引起Chl含量下降,Chla／b比值上升;③上述这些生理反应与果树的抗热性有关,可用于果树的抗热性鉴定。     

1982-2018年中国植被覆盖变化非线性趋势及其格局分析

探究植被覆盖变化是评估陆地生态系统环境变化的重要手段,但现有研究多采用线性趋势来表达植被覆盖的变化情况而忽略了趋势的非线性。本文使用GLASS FVC数据,利用BFAST方法和格局分析,探讨了1982-2018年我国植被覆盖变化的非线性趋势及其分布格局。结果表明：（1）与线性趋势方法的对比发现,BFAST的检测结果揭示了四川盆地、黄土高原等地的植被覆盖显著增加趋势其实存在中断,青海和东北等地植被覆盖经历了由退化到改善的过程而并非简单的线性增加,而青藏高原中东部等地则由原先的改善趋势变为了退化趋势。（2）将非线性趋势结果进行分类,其中单调型增加类型占比最多,达到33.58%,主要分布在内蒙古、陕西及河南等地;单调型减少占比1.82%,主要分布在东南沿海地区;中断型增加占比22.91%,主要分布在四川盆地东部和华北地区;中断型减少占比2.68%,主要分布在青藏高原东南部;由增到减占比4.20%,主要分布在青海等地;由减到增占比14.62%,主要分布在吉林等地。大范围的植被覆盖增加趋势充分反映了我国过去几十年植被的改善,但同时存在的减少趋势表明潜在的植被退化风险仍不可忽视。（3）不同趋势类型发生改变的时间有所差异,总体上1988-1999年间发生的改变较少,而2000-2011年间发生的改变较多,我国21世纪以来实施的大规模生态保护和恢复工程对植被的改善过程有重要影响。（4）分布格局上,植被覆盖改善趋势类型（单调型增加,中断型增加,由减到增）呈现大聚集,小分散的特点,具有复杂的形状;退化趋势类型（单调型减少,中断型减少,由增到减）的面积均较小,分布也相对离散。全国尺度上趋势空间格局呈现一定规律但分布的异质性较大,区域尺度上植被覆盖经受的干扰显著,变化过程实际也是较为复杂的。本研究表明,使用非线性趋势方法和格局分析,可以更准确地评估植被覆盖的时空变化,从而为生态环境相关工作的开展提供科学的参考。     

辣椒开花结果期对干旱胁迫的形态与生理响应

在遮雨网室选用抗旱性较强的农城椒二号和抗旱性较弱的陕蔬2001,研究辣椒在轻度、中度和重度干旱胁迫下不同时间的生长、产量、渗透调节物质、保护酶活性的变化规律及其生理调节机制。结果表明:随干旱胁迫时间的延长,辣椒的株高、分枝数、叶面积、单位面积产量、叶绿素含量和叶片相对含水量的抗旱系数呈下降趋势,下降速率与干旱胁迫程度呈正相关,与品种的抗旱性呈负相关;脯氨酸、丙二醛含量和细胞膜透性相对值随干旱胁迫时间的延长呈上升趋势;POD、SOD、CAT活性和可溶性蛋白相对值随着干旱胁迫时间的延长先升高后下降,抗旱性强的材料增加幅度低于抗旱性弱的材料;可溶性糖含量的相对值在轻度和中度干旱胁迫下呈上升趋势,在重度干旱胁迫下呈上升—下降趋势,且抗旱性强的材料上升速度大于抗旱性弱的材料。相关分析表明,干旱胁迫下,产量与株高、分枝数、叶片叶绿素含量、叶面积、叶片相对含水量抗旱系数呈显著正相关;与细胞膜透性、CAT活性和可溶性蛋白含量抗旱系数呈显著负相关。主成分分析表明,用作辣椒抗旱性鉴定的主要指标是单株产量、株高、叶面积、分枝数、可溶性蛋白、可溶性糖、MDA、叶绿素含量和细胞膜透性及叶片相对含水量,叶片POD、SOD、CAT活性、脯氨酸含量可做为辣椒抗旱性鉴定的次要鉴选指标。