激光照射对角膜蛋白质二级结构影响的光谱分析

为了研究激光照射后角膜蛋白质化学组成的改变,探讨激光角膜损伤的发生机制,将日本大耳白兔随机分为正常对照组和激光损伤组,选取角膜上皮层和基质层作为研究对象,通过傅里叶变换红外光谱技术(Fou-rier transform infrared spectroscopy,FT-IR)对角膜组织酰胺I带中蛋白质二级结构各吸收峰进行定量分析,观察激光照射后蛋白质分子结构的改变情况。结果显示,激光照射后出现蛋白质二级结构吸收峰的位移和积分百分比的改变。激光照射可使角膜上皮层和基质层蛋白质构象发生改变,从而导致蛋白质结构稳定性下降和蛋白质生物功能的破坏。     

不同能量激光照射后中华大蟾蜍早期胚胎发育及其蛋白质谱变化的研究

应用YAG倍频激光器选择不同能量密度激光微束对中华大蟾蜍4细胞期胚胎进行照射,观察胚胎的生长、发育状况,统计出膜率、畸胎率和死亡率。实验结果表明：低能量激光照射明显地提高了胚胎的孵化速度,且使畸变率降低;而高能量激光对胚胎发育有明显抑制作用,而且使畸变率增高。不同能量密度激光微束照射中华大蟾蜍早囊胚,0．5h后制备样品进行双向电泳,凝胶成像后用PDQuest软件进行分析。结果显示激光照射对蛋白质点数及其分布有影响,影响程度与照射激光密度有关。结论：激光照射可以影响胚胎发育和蛋白质的合成。     

不同激光照射方法对周围神经再生的影响

为研究不同半导体激光照射方法对周围神经损伤的影响,将96只家兔随机分为3周,6周,9周,12周4个观察期组,每个观察期组又随机分为不同照射方法的治疗组和对照组。建立动物模型后,各照射组在术后1d开始照射治疗,激光功率为10mw,每次照射10rain,每天一次,连续照射10d。照射治疗A组对准损伤神经吻合部位进行照射,照射治疗B组照射家兔L5、L6脊髓节段,照射治疗c组在对准吻合处进行照射同时还要照射L5、L6脊髓节段,对照组激光输出功率为零。实验结果表明低能量半导体激光照射能促进轴突再生,改善再生神经功能,以同时照射损伤周围神经部位和相应脊髓节段效果最为显著。     

Nd：YAG激光对提高根管治疗术效果的作用机理探讨

用波长1.06μm,输出功率20W的Nd:YAG激光照射根管壁后,常规根管糊剂＋牙胶尖侧压法充填,对照组,直接根管填。通过测量酚醛树脂在根管模截面中所占面积比值和染料渗入长度,发现实验组和对照组之间无明显差异（P＞0.05）。认为单纯激光照射对根尖封闭能力的影响并不明显。激光照射在根管治疗术中的作用可能是通过其它途径。     

激光联合环磷酰胺对荷瘤鼠脾重和外周免疫细胞数量影响的试验研究

目的:试验以小鼠S180腹水瘤为动物实验模型,对He-Ne激光照射与化疗药物联合应用对S180荷瘤鼠脾脏重量和免疫细胞数量影响作了系统性研究。方法:应用11.00J/cm2,14.67J/cm2和22.00J/cm2三种剂量He-Ne激光照射荷瘤鼠内眼角,同时联合应用化疗药物环磷酰胺,以观察其对荷瘤鼠脾脏重量和免疫细胞数量的影响。结果:健康小鼠脾脏随日龄增加呈持续上升趋势,肿瘤对照组及CYT对照组在第4d.显著升高。此后便持续下降,尤其是CYT对照组,第8d.荷瘤鼠脾脏萎缩极为严重。而CYT/He-Ne激光联合应用组小鼠脾脏在第4d.轻微下降后,便呈持续上升趋势;CYT对外周血白细胞总数、淋巴细胞数和中性幼稚细胞数有着显著的抑制作用(P<0.01,P<0.05)。CYT/He-Ne激光联合应用组白细胞总数和中性幼稚细胞数的变化较单纯CYT应用组为轻微。结论:He-Ne激光对CYT及肿瘤本身所致免疫抑制具有明显的改善效应。     

Nd∶YAG激光对提高根管治疗术效果的作用机理探讨

用波长1.06μm,输出功率20W的Nd∶YAG激光照射根管壁后,常规根管糊剂+牙胶尖侧压法充填。对照组,直接根管充填。通过测量酚醛树脂在根管横截面中所占面积比值和染料渗入长度,发现实验组和对照组之间无明显差异(P>0.05)。认为单纯激光照射对根尖封闭能力的影响并不明显。激光照射在根管治疗术中的作用可能是通过其它途径。     

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.     

Er,Cr:YSGG激光照射对牙周病牙根面的影响

目的:观察不同能量输出的Er,C r:YSGG激光照射对牙周病牙根面的影响,探讨Er,C r:YSGG激光用于牙周治疗的可行性和有效性。方法:选择因牙周病松动Ⅲo而拔除牙12颗,随机分3组,每组4颗。分别以1.5 W、2.0 W、2.5 W激光照射。扫描电镜(SEM)观察各组牙根表面和纵剖面。结果:低倍镜下各组根面改变无明显差异,表面干净,粗糙程度均匀,未见明显弹坑状痕迹;高倍镜下1.5 W激光照射组显示根面粗糙程度更均匀一致,无弹坑状改变,无熔融、炭化迹象,未见牙本质小管暴露。结论:Er,C r:YSGG激光能有效去除牙周病牙根面玷污层及病变牙骨质,根面无熔融、炭化等改变。1.5 W激光照射组优于2.0 W组和2.5 W组。     

810nm弱激光照射对大鼠急性脊髓损伤后巨噬细胞极化的作用研究

本研究构建急性大鼠脊髓夹伤模型,并将大鼠随机分为单纯脊髓损伤对照组及脊髓损伤联合弱激光照射组。照射组应用810 nm波长,150 m W照射功率,照射光斑0.3 cm^2的弱激光对脊髓损伤区进行经皮照射,连续照射3天,7天或14天。应用免疫荧光、免疫印迹实验方法,测定脊髓损伤区巨噬细胞及小胶质细胞的极化表达。应用酶联免疫吸附法测定脊髓损伤区白细胞介素4的表达情况。应用坚牢蓝髓鞘染色测定两组损伤脊髓中髓鞘保留的差异。采用BBB评分对两组大鼠后肢运动功能的恢复进行评估。结果表明,810 nm弱激光对脊髓损伤区连续照射3天,7天后,可显著减少M1型巨噬细胞及其标志物诱导型一氧化氮合酶的表达,在7天时间增加M2型巨噬细胞及其标志物精氨酸酶1的表达。弱激光照射组白细胞介素4的表达明显增加。损伤后14天,弱激光照射组脊髓损伤区髓鞘保留面积比值明显提高。损伤后7天及14天时,弱激光照射组大鼠的BBB评分明显升高。该实验结果表明,810 nm弱激光经皮照射,可增加大鼠急性脊髓损伤区M2型巨噬细胞及小胶质细胞的表达,并减少脊髓损伤后的髓鞘脱失,促进脊髓损伤大鼠运动功能的恢复。     

手外伤激光照射康复护理观察研究

目的：探讨激光照射对手外伤患者的康复效果。方法：随机选取87例手外伤患者为观察组,采用激光照射与作业疗法,支具、心理疗法相结合;对照组63例,只采用作业疗法、支具及心理疗法。观测伤部疼痛减轻及消退情况,肿胀消退时间,康复治疗2月进行手功能评定。结果：观察组疼痛消失时间,肿胀消退时间,手功能康复均优于对照组,两组比较,差异均具有显著意义。结论：手外伤术后患者激光照射治疗能明显缩短手康复时间,提高康复质量。     

Changes induced by oxygen in rat liver proteins identified by high-resolution two-dimensional gel electrophoresis.

Molecular oxygen (O2) regulates the expression of a variety of genes. Several of the proteins that respond to changes in oxygen concentration have been identified in a variety of cell lines. We extend these previous studies by analyzing the effect of oxygen on the entire protein expression profile of an intact organ using high-resolution two-dimensional gel electrophoresis. To this end, we used an isolated, in vitro perfused organ preparation to produce two groups of rat livers perfused with high (95% O2, 5% CO2) or low (95% N2, 5% CO2) oxygen concentrations. Using two-dimensional gel electrophoresis we compared the protein expression profiles of both groups of livers. Computer analysis of the files obtained after laser densitometry of the two-dimensional gels revealed two spots that were strongly up-regulated in high PO2 perfused livers compared with low PO2 perfused livers. These spots were analyzed by peptide mass fingerprinting analysis. These spots were identified as arginase 1 (liver-type arginase; EC 3.5.3.1) and mitochondrial enoyl-CoA hydratase 1 (EC 4.2.1.17). The possible role of these proteins in its new context of oxygen availability is discussed.     

Gel-based proteomics of unilateral irradiated striatum after gamma knife surgery

Gamma knife surgery (GKS) is used for the treatment of various brain disorders. The biological effects of focal gamma ray irradiation on targeted or surrounding areas in the brain are not well-known. In the present study, we evaluated protein expression changes in the unilateral irradiated (60 Gy) striatum in rat. Striata of irradiated and control brains were dissected 16 h post-irradiation for analysis by large-format two-dimensional gel electrophoresis (2-DGE). In parallel, we also examined the un-targeted contralateral striatum over the control for potential changes in proteins patterns that may have occurred due to the effects of irradiation to the unilateral striatum. A total of 17 reproducible and differentially expressed silver nitrate-stained protein spots in the irradiated striatum was detected on 2-D gel. Their subsequent analysis by tandem mass spectrometry (nESI-LC-MS/MS) resulted in the identification of 13 nonredundant proteins. Interestingly, out of these 13 changed proteins, 2 proteins were also detected in the contralateral striatum. Some of the significantly changed proteins identified were creatine kinase, protein disulfide isomerase A3 precursor (PDA3), and peroxiredoxin 2 (Prx2). Western analysis with anti-PDA3 and anti-Prx2 antibodies revealed 4 and 2 cross-reacting protein spots on 2-D gel blots. Interestingly, after GKS, in the irradiated and un-irradiated striata, these spots showed a shift toward the acidic side, suggesting post-translational modifications. Taken together, these results indicate that unilateral irradiation during GKS triggers molecular changes in the bilateral striata.     

Fluorescence two-dimensional difference gel electrophoresis and mass spectrometry based proteomic analysis of Escherichia coli

Separation and relative quantitation of complex protein mixtures remain two of the most challenging aspects of proteomics. Here an advanced technique called fluorescence difference 2-D gel electrophoresis technology (2D-DIGE) has been applied to a model system study of the Escherichia coli proteome after benzoic acid treatment. The molecular weight and charge matched cyanine dyes enable pre-electrophoretic labelling of control and treated samples which are then mixed and run in the same gel. Pooled control and treated samples labelled with Cy trade mark 3 were used as an internal standard for both Cy5 labelled control and treated E. coli samples. Together with DeCyder trade mark imaging analysis software, more accurate quantitative analysis than conventional two-dimensional polyacrylamide gel electrophoresis was achieved. Using matrix-assisted laser desorption/ionization-time of flight and quadrupole-time of flight mass spectrometry a total of 179 differentially expressed protein spots were identified. These included enzymes, stress related and substrate (e.g. amino acids, maltose, ribose and TRP repressor) binding proteins. Of the spots analysed, 77% contained only one protein species per spot, hence the change in protein expression measured was solely attributed to the identified protein. Many membrane proteins and protein isoforms were identified indicating both adequate solubilization of E. coli samples and potential post-translational modification. The results indicate that the regulatory mechanisms following benzoic acid treatment of E. coli are far more complicated than hitherto expected.     

胶质芽胞杆菌对磷矿石风化作用时细菌蛋白质的双向电泳分析

本文研究对磷矿石有风化作用的胶质芽胞杆菌(Bacillus muscilaginosus)在风化磷矿石过程中菌体蛋白质组的变化及差异, 以此了解该风化作用的分子生物学机制。将该菌接入液体培养基中, 加磷矿粉为处理组, 不加磷矿粉为对照组, 静置培养。培养10 d 后, 取上清液离心后的固体物和部分上清液中的外分泌蛋白用作菌体蛋白质组学分析。菌体蛋白质组二维电泳图谱进行比较和软件分析后, 处理组可见蛋白点数为(1134±98)个, 对照组可见蛋白点数为(729±53)个, 两者比较具有统计学差异(P<0.0     

失血性休克大鼠肝脏蛋白质的差异分析

 应用蛋白质双向凝胶电泳 (two-dimensional polyacrylamide gel electrophoresis, 2-DE) 技术,分析在急性重度失血性休克 (refractory hemorrhagic shock, RHS) 条件下,大鼠肝脏蛋白质组表达的差异.16 只雄性 Wistar 大鼠随机分成正常对照组 (sham hemorrhage shock, SHS) 和 RHS 模型组,每组 8 只.采用股动脉放血的方法制备模型,在规定时间内处死大鼠并分离肝脏,提取肝脏总蛋白质后进行 2-DE.运用 Image Master 2D Platinum v 5.0 凝胶图像分析软件对 2-DE 凝胶图像进行差异表达分析.有意义的差异蛋白质点用基质辅助激光解析电离飞行时间质谱进行肽质量指纹图谱分析,借助 Swiss-prot 数据库进行蛋白质搜索和鉴定.SHS 组和 RHS 组肝脏的 2-DE 图谱,分别平均识别到 698±11 和 700±13 个蛋白质点,SHS 组和 RHS 组肝脏间平均匹配率达88%～92 %.共发现 10 个差异有意义的蛋白质点,鉴定出了肿瘤抑制性抗原gp96、葡萄糖调节蛋白58、过氧还蛋白Ⅰ、细胞色素b5、谷胱甘肽转移酶、ATP合酶β亚单位、二磷酸果糖酶 B、三磷酸甘油醛脱氢酶等8种蛋白质.结果表明,以 2-DE 技术得到重复性和分辨率都较好的 2-DE图谱,并初步鉴定急性重度失血性休克后大鼠肝脏的差异表达蛋白质,为深入研究失血性休克的生理病理机制及寻找失血性休克预防和治疗的生物标志物提供了依据.     

A proteomic analysis of allograft rejection in rats after liver transplantation

In order to understand the allograft rejection in orthotopic liver transplantation (OLT), an allograft rejection rat model was established and studied by proteomic approach. The protein expression profiles of liver tissues were acquired by fluorescence two-dimensional difference gel electrophoresis (2D DIGE) that incorporated a pooled internal standard and reverse fluorescent labeling method. The expression levels of 27 protein spots showed significant changes in acute rejection rats. Among these spots, 19 were identified with peptide mass fingerprinting using matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF MS) after tryptic in-gel digestion. The results of the present paper could be helpful for our better understanding of allograft rejection in organ transplantation.     

Proteomic signatures for histological types of lung cancer

We performed proteomic studies on lung cancer cells to elucidate the mechanisms that determine histological phenotype. Thirty lung cancer cell lines with three different histological backgrounds (squamous cell carcinoma, small cell lung carcinoma and adenocarcinoma) were subjected to two-dimensional difference gel electrophoresis (2-D DIGE) and grouped by multivariate analyses on the basis of their protein expression profiles. 2-D DIGE achieves more accurate quantification of protein expression by using highly sensitive fluorescence dyes to label the cysteine residues of proteins prior to two-dimensional polyacrylamide gel electrophoresis. We found that hierarchical clustering analysis and principal component analysis divided the cell lines according to their original histology. Spot ranking analysis using a support vector machine algorithm and unsupervised classification methods identified 32 protein spots essential for the classification. The proteins corresponding to the spots were identified by mass spectrometry. Next, lung cancer cells isolated from tumor tissue by laser microdissection were classified on the basis of the expression pattern of these 32 protein spots. Based on the expression profile of the 32 spots, the isolated cancer cells were categorized into three histological groups: the squamous cell carcinoma group, the adenocarcinoma group, and a group of carcinomas with other histological types. In conclusion, our results demonstrate the utility of quantitative proteomic analysis for molecular diagnosis and classification of lung cancer cells.     

Analysis of the proteome in the human pituitary

The pituitary is the master endocrine gland responsible for the regulation of various physiologic and metabolic processes. Proteomics offers an efficient means for a comprehensive analysis of pituitary protein expression. This paper reports on the application of proteomics for the mapping of major proteins in a normal (control) pituitary. Pituitary proteins were separated by two-dimensional gel electrophoresis with immobilized pH 3-10 gradient strips. Major protein spots that were visualized in the two-dimensional gel by silver staining were excised, and the proteins in these spots were digested with trypsin. The tryptic digests were analyzed by mass spectrometry, and the mass spectrometric data were used to identify the proteins through searches of the SWISS-PROT or NCBInr protein sequence databases. The majority of the proteins were identified on the basis of peptide mass fingerprinting data obtained by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Several proteins were also characterized based on product-ion spectra measured by post-source decay analysis and/or liquid chromatography-electrospray-quadrupole ion trap mass spectrometry. To date, 62 prominent protein spots, corresponding to 38 different proteins, were identified. The identified proteins include important pituitary hormones, structural proteins, enzymes, and other proteins. The protein identification data were used to establish a two-dimensional reference database of the human pituitary, which can be accessed over the Internet (http://www.utmem.edu/proteomics). This database will serve as a tool for further proteomics studies of pituitary protein expression in health and disease.     

595nm 激光对兔耳瘢痕成纤维细胞增殖活性的影响

目的：探讨兔耳增生性瘢痕动物模型形成过程中成纤维细胞增殖活性的动态变化及595nm Vbeam激光照射的影响。方法：利用兔耳腹侧面建立增生性瘢痕模型,按不同时间段取材,采用免疫组织化学方法观察瘢痕形成不同时期增殖细胞核抗原（PCNA）蛋白的表达,对比研究在兔耳增生性瘢痕形成过程中595nm Vbeam激光照射对成纤维细胞增殖活性的影响。结果：上皮化后增殖细胞核抗原阳性细胞反应率随时间推移逐渐增高,至上皮化后4周达高峰。从上皮化后1周开始即可见到595nm Vbeam激光照射后增殖细胞核抗原蛋白表达较对照组明显减弱,成纤维细胞增殖活性受到明显抑制,两组差异具有极显著性（P〈0．01）。结论：595nm Vbeam激光照射可明显抑制兔耳增生性瘢痕成纤维细胞的增殖活性,对兔耳腹侧面增生性瘢痕有明显的防治作用。