甲蓝

The neural complex of ascidian Styela clava was observed by two histological methods, and the methionine-enkephalin in the neural complex of Styela clava was analyzed by the immunohistochemical S-ABC (strept avidin-biotin complex) technique,respectively.

采用甲苯胺蓝染色和Golgi镀银法对柄海鞘神经复合体的结构进行显微观察，并通过免疫组织化学方法对甲硫氨酸脑啡肽在神经复合体中的分布进行了研究。

In the article, distributions of mast cells in spleen, cecum tonsilla and cecum of francolin were observed by improved toluidine dye method,alcian blue-Safranine dye method,long time toluidine dye method.

应用阿尔新蓝-藏花红染色法、改良甲苯胺蓝染色法、长效甲苯胺蓝染色法等三种不同染色方法观察鹧鸪脾脏、盲肠、盲肠扁桃体肥大细胞分布情况，结果表明：1。

Results Results: After intensie mucus remoal, high-quality images were obtained using methylene blue and toluidine blue.

结果 去除染色液之后，用亚甲基蓝和甲苯胺蓝染色的组织获得了高质量的图像。

The screening was carried out by single strand conformational polymorphorsim : the PCR product precipitation was dissolved in 20μl denaturing and loading buffer (95%+20mMEDTA+ 0.05% bromophenol blue+0.05% xylene cyanol), heat-denatured in 95 ℃ to two single DNA strands.

通过单链构象多态性分析筛查RDS基因突变；沉淀回收的PCR产物以20μl变性上样液（95%甲酰胺+20mMEDTA+0.05%溴酚蓝+0.05%二甲苯蓝）溶解后热变性为两条DNA单链后，上样于8%非变性聚丙烯酰胺凝胶（含5%甘油），700伏电压、4℃电泳16小时。

Phycobiliprotein are important light-harvesting pigment proteins in Cyanophyceae, Rhodophyceae, Cryptophyceae and a little Pyrrophyceae.

中文题名极大螺旋藻藻蓝蛋白基因的克隆及其表达研究副题名外文题名 Cloning and sequencing of the phycocyanin gene from Spirulina maxima and its over-expression in Pichia pastoris 论文作者于平导师岑沛霖学科专业生物化工研究领域\研究方向学位级别博士学位授予单位浙江大学学位授予日期2003 论文页码总数110页关键词藻胆蛋白基因克隆藻蓝蛋白基因螺旋藻馆藏号BSLW /2003 /Q949 /32 藻胆蛋白是蓝藻、红藻、隐藻和一些少数甲藻中的一种重要的捕光色素蛋白。

Primary cultured chondrocytes are poly-angle, cytoplasm-rich, and their nuclei are either round or oval with clear necleole. Metachromatic and alcian blue positive staining in primary cultured chondrocytes was observed. Intercellular matrix was anti-collagen type Ⅱ staining but not anti-collagen type Ⅰ staining by IHC assay.

原代培养软骨细胞呈多角型，胞质丰富，胞核成圆形或椭圆型，核仁清楚，甲苯胺蓝呈异染性，阿尔新蓝8Gx 染色阳性，细胞外基质Ⅱ型胶原免疫组化染色阳性，Ⅰ型胶原染色阴性。

Discipline, animal husbandry, clothing sets some magic back to the Blue A, BUFF steal the scene to see a range of control is very important, as far as possible do not eat even a Blast Firelight, beat ice ring flash disappear silent dining, or wake up the sheep out to see who did not blue, but not killed, just not to be killed and would not lose on win sooner or later.

戒律牧，穿一些回蓝套魔甲，偷BUFF看情况定，射程控制很重要，尽量不吃震爆心火一套连，打不过冰环沉默闪现消失吃喝，或者羊掉唤醒，看谁先没蓝，不求打死，只求不被打死，不会输，就早晚能赢。

Study in vitro The chondrocytes of SD rat were isolated from epiphyses by collagenase Ⅱ and the cultured chondrocytes were confirmed by HE, toluidine blue, alcian blue staining, and by IHC with anti-collagen type Ⅰ and Ⅱ.

体外实验 SD大鼠骺软骨细胞分离培养，HE、甲苯胺蓝、标准阿尔新蓝染色及Ⅰ、Ⅱ免疫组化染色鉴定，绘制细胞生长曲线。

The effects and mechanism of GABAergic neurons, NOergic neurons, opioid peptide and cyclic adenosine monophosphate in the nucleus reticularis thalami on sleep-wakefulness cycle of rats and the effects and mechanism of the 5-HTergic nerve fibers project from the nucleus raphes dorsalis to RT on sleep-wakefulness cycle of rats were investigated with the methods of brain stereotaxic, nucleus spile, microinjection and polysomngraphy.1. The effects of GABAergic neurons in RT on sleep-wakefulness cycle of rats1.1 Microinjection of 3-mercaptopropionic acid (3-MP, a kind of glutamate decarboxylase inhibitor) into RT. On the day of microinjection, sleep only decreased a litter. On the second day, sleep marked decreased and wakefulness marked increased. On the third and fourth day, sleep and wakefulness stages resumed to normal.1.2 Microinjection of gamma-amino butyric acid (GABA 1.0μg) into RT enhanced sleep and reduced wakefulness compared with control; while microinjection of L-glutamate (L-Glu, 0.2μg) decreased sleep and increased wakefulness; microinjection of bicuculline (BIC, 1.0μg), a GABAA receptor antagonist, enhanced wakefulness and reduced sleep; microinjection of baclofen (BAC, 1.0μg), GABAB receptor agonist, had the same effects as GABA.2. The effects of NOergic neurons in RT on sleep-wakefulness cycle of rats2.1 Microinjection of L-arginine (L-Arg, 0.5μg) into RT decreased sleep compared with control, but there were on statistaical difference between L-Arg group and control; while microinjection of sodium nitroprusside (SNP, 0.2μg), a NO donor into RT, sleep marked decreased and wakefulness marked increased. Microinjection of nitric oxide synthase inhibitor, N-nitro-L-arginine (L-NNA, 2.0μg) into RT enhanced sleep and reduced wakefulness.2.2 After simultaneous microinjection of L-NNA (2.0μg) and SNP (0.2μg) into RT, SNP abolished the sleep-promoting effect of L-NNA compared with L-NNA group; after simultaneous microinjection of L-NNA (2.0μg) and L-Arg(0.5μg) into RT, we found that L-NNA could not blocked the wakefulness-promoting effect of L-Arg.3. The effects of opioid peptide in RT on sleep-wakefulness cycle of rats3.1 Microinjection of morphine sulfate (MOR, 1.0μg) into RT increased wakefulness and decreased sleep compared with control; while microinjection of naloxone hydrochloride (NAL, 1.0μg), the antagonist of opiate receptors, into RT, enhanced sleep and reduced wakefulness.3.2 After simultaneous microinjection of MOR (1.0μg) and NAL (1.0μg) into RT, the wakefulness-promoting effect of MOR and the sleep-promoting effect of NAL were not observed compared with control.4. The effects of cAMP in RT on sleep-wakefulness cycle of rats Microinjection of cAMP (1.0μg) into RT increased sleep and decreased wakefulness compared with control; microinjection of methylene blue (MB,1.0μg) into RT enhanced sleep and reduced wakefulness compared with control.5. The effects of the 5-HTergic nerve fibers project from DRN to RT on sleep-wakefulness cycle of rats5.1 When L-Glu (0.2μg) was microinjected into DRN and normal sodium (NS,1.0μg) was microinjected into bilateral RT. We found that sleep was decreased and wakefulness was increased compared with control; when L-Glu (0.2μg) was microinjected into DRN and methysergide (MS,1.0μg), a non-selective 5-HT antagonist, was microinjected into bilateral RT, We found that sleep was enhanced and wakefulness was reduced compared with L-Glu group.5.2 When p-chlorophenylalanine (PCPA, 10μg) was microinjected into DRN and NS (1.0μg) was microinjected into bilateral RT, We found that sleep was increased and wakefulness was decreased compared with control; microinjection of 5-hydroxytryptaphan (5-HTP, 1.0μg), which can convert to 5-HT by the enzyme tryptophane hydroxylase and enhance 5-HT into bilateral RT, could block the effect of microinjection of PCPA into DRN on sleep-wakefulness cycle.

本研究采用脑立体定位、核团插管、微量注射、多导睡眠描记等方法，研究丘脑网状核(nucleus reticularis thalami，RT)中γ-氨基丁酸(gamma-amino butyric acid ，GABA)能神经元、一氧化氮(nitrogen monoxidum，NO)能神经元、阿片肽类神经递质、环一磷酸腺苷(cyclic adenosine monophosphate，cAMP)及中缝背核(nucleus raphes dorsalis，DRN)至RT的5-羟色胺(5-hydroxytryptamine，5-HT)能神经纤维投射对大鼠睡眠-觉醒周期的影响及其作用机制。1 RT内GABA能神经元对大鼠睡眠-觉醒周期的影响1.1大鼠RT内微量注射GABA合成关键酶抑制剂3-巯基丙酸(3-MP，5μg)，注射当天睡眠时间略有减少，第二日睡眠时间显著减少，觉醒时间明显增多，第三、四日睡眠和觉醒时间逐渐恢复至正常。1.2大鼠RT内微量注射GABA受体激动剂GABA( 1.0μg)后，与生理盐水组比较，睡眠时间增加，觉醒时间减少；而RT内微量注射L-谷氨酸(glutamic acid， L-Glu， 0.2μg)后，睡眠时间减少，觉醒时间增加；RT内微量注射GABAA受体阻断剂荷包牡丹碱(bicuculline，BIC，1.0μg)后，睡眠时间减少，觉醒时间增加；RT内微量注射GABAB受体激动剂氯苯氨丁酸(baclofen，BAC，1.0μg)后，产生了与GABA相似的促睡眠效果。2 RT内NO能神经元对大鼠睡眠-觉醒周期的影响2.1大鼠RT内微量注射NO的前体L-精氨酸(L-Arg，0.5μg)后，与生理盐水组对比，睡眠时间略有减少，但无显著性意义；而RT内微量注射NO的供体硝普钠(Sodium Nitroprusside，SNP，0.2μg)后可明显增加觉醒时间，缩短睡眠时间；微量注射一氧化氮合酶抑制剂L-硝基精氨酸(L-arginine，L-NNA，2.0μg)后，引起睡眠时间增多，觉醒时间减少。2.2大鼠RT内同时微量注射L-NNA(2.0μg)和SNP(0.2μg)后与L-NNA组比较发现SNP逆转了L-NNA的促睡眠作用；RT内同时微量注射L-NNA(2.0μg)和L-Arg(0.5μg)后，与L-NNA(2.0μg)组比较发现L-Arg可以增加觉醒而缩短睡眠，其促觉醒作用未能被NOS的抑制剂L-NNA所逆转。3 RT内阿片肽对大鼠睡眠-觉醒周期的影响3.1大鼠RT内微量注射硫酸吗啡(morphine sulfate，MOR，1.0μg)后与生理盐水组对比，睡眠时间减少而觉醒时间增加； RT内微量注射阿片肽受体拮抗剂盐酸纳洛酮(naloxone hydrochloride，NAL，1.0μg)后与生理盐水组比较，睡眠时间增加而觉醒时间减少。3.2大鼠RT内同时微量注射MOR(1.0μg)和NAL(1.0μg)后，与生理盐水组对比，原有的MOR促觉醒效果和NAL的促睡眠效果都没有表现。4 RT内环一磷酸腺苷信使对大鼠睡眠-觉醒周期的影响大鼠RT内微量注射cAMP(1.0μg)后与NS(1.0μg)组比较，睡眠时间增多而觉醒时间减少；RT内微量注射亚甲蓝(methylene blue，MB，1.0μg)后，与NS组比较，睡眠时间增多而觉醒时间减少。5中缝背核投射到丘脑网状核的5-羟色胺能神经纤维对大鼠睡眠-觉醒周期的影响5.1大鼠DRN内微量注射L-Glu(0.2μg)，同时在双侧RT内微量注射NS (1.0μg)后，与对照组(DRN和双侧RT注射NS， 0.2μg)比较，睡眠时间减少，觉醒时间增多；大鼠DRN内微量注射L-Glu(0.2μg)，同时在双侧RT内微量注射二甲基麦角新碱(methysergide， MS， 1.0μg )后，与对照组(DRN注射L-Glu 0.2μg，双侧RT注射NS 1.0μg)比较，睡眠时间增多，觉醒时间减少。5.2大鼠DRN内微量注射对氯苯丙氨酸(p-chlorophenylalanine，PCPA，10μg)，同时在双侧RT内微量注射NS (1.0μg)后，与对照组(DRN和双侧RT注射NS， 1.0μg)比较，睡眠时间增多，觉醒时间减少；大鼠DRN内微量注射PCPA(10μg)，产生睡眠增多效应后，在双侧RT内微量注射5-羟色胺酸(5-hydroxytryptaphan ， 5-HTP， 1.0μg )后，与对照组(DRN注射PCPA 10μg，双侧RT注射NS 1.0μg)比较，睡眠时间减少，觉醒时间增多。

objective to investigate staining methods for two-dimensional gel(2-de)electrophoresis in multidrug resistance of gastric cancer.methods cultured vincristine-resistant human gastric cancer cell line sgc7901/vcr and its parental cell line sgc7901.variant amount protein of those cells were separated by 2-de.gels were stained with silver nitrate or colloidal coomassie brilliant blue,and scanned by image scanner.results well-resolved,reproducible 2-de patterns of sgc7901/vcr and sgc7901 were established.silver staining was better when protein sample amount was low,overloaded protein will interfere resolution of the maps.gels stained with colloidal coomassie brilliant blue had more protein spots numbers and abundance without apparent trails when increased loading protein sample.conclusion two staining methods were influenced largely by the sum of protein samples,properly selection may be helpful for further study with proteomics in multidrug resistance of gastric cancer.

低甲氧基果胶的胶凝机理及防止预凝胶。。。他扎罗汀凝胶与克林霉素凝胶治疗痤疮。。。注射隆乳后经乳晕切口取出聚丙烯酰胺。。。目的分析利用蛋白质组学方法研究胃癌耐药相关蛋白质中双向电泳凝胶的染色显示。方法培养胃癌细胞sgc7901和长春新碱诱导的耐药胃癌细胞sgc7901/vcr，用双向凝胶电泳技术分离总蛋白，银染及胶体考马斯亮蓝染色，image scanner扫描仪扫描凝胶。结果获得了背景清晰、重复性好的双向凝胶电泳图谱，两种染色凝胶相比，硝酸银染色在样品少时显示更佳，过量则影响图像质量，而胶体考马斯亮蓝染色在上样量增加时的凝胶蛋白质点数目及丰度均增加，并无明显拖尾。结论两种显色方法受样品量影响较大，恰当选用有利于通过蛋白质组学研究胃癌耐药机制工作的进一步开展。

The split between the two groups can hardly be papered over.

这两个团体间的分歧难以掩饰。

This approach not only encourages a greater number of responses, but minimizes the likelihood of stale groupthink.

这种做法不仅鼓励了更多的反应，而且减少跟风的可能性。