Study on Fluorimetry for Determination of Phenylalanine Using Enzyme Substituted by the UV-Light

在碱性介质中， 光子可以取代苯丙氨酸氧化酶使苯丙氨酸分解并生成H2 O2 ， 然后用氯化血红素作为模拟酶检测H2O2 ， 从而建立间接测定苯丙氨酸的新方法.In alkaline medium,the photon can substitute phenylalanine oxydase,and cause phenylalanine to decompose and produce H 2 O 2 ,which are detected by using HPA to be coupled and produce strong fluorescent substance in present of mimetic peroxydase-hemin.Thus,a new fluorimetry for the determination of phenylalanine was developed.福建省自然科学基金!（B96003

p38MAPK induces apoptosis of glioma cell

目的　研究 p38MAPK基因转染大鼠胶质瘤细胞系C6后对其生物学特性的影响 .方法　利用脂质体介导法将p38MAPK基因导入大鼠胶质瘤细胞系 C6中 ,用免疫细胞化学染色检测其在细胞转染前后的表达情况 ,用 HE染色、流式细胞仪等方法研究其对细胞形态、粘着状况和生长周期的影响 .结果　转染 p CMV5 - p38MAPK质粒组 p38MAPK蛋白表达阳性 ,细胞形态发生变化 ,贴壁性降低 ,出现大量凋亡细胞 .结论　转染 p38MAPK基因可诱导胶质瘤细胞凋

Regulation of Cell Cycle of Glioma C6 Cells by Regulator of G Protein Signaling 16

目的　探讨G蛋白调节子 16(RGS16)对胶质瘤C6细胞周期的影响。方法　利用脂质体介导法将RGS16基因导入C6细胞中 ,在倒置显微镜下观察细胞形态变化和贴壁生长情况 ;免疫细胞化学法检测转染前后RGS16蛋白的表达情况 ;流式细胞仪检测转染pCMV 5 RGS16和 pCMV 5质粒后每隔 12h后的细胞周期变化。 结果　转染pCMV 5 RGS16质粒 2 4h后 3 0 .0 %细胞贴壁性降低 ,突起收缩 ,细胞变圆 ,72h之后细胞又恢复正常 ;RGS16蛋白的表达呈时相性 ,3 6h时表达率最高 (阳性率为13 .0 % ) ,72h表达终止 ;C6细胞的各期细胞比例变化与RGS16蛋白表达对应 ,在 3 6h时G1期比例从转染前的 70 .5 %降低到60 .2 % ,S期比例从 2 0 .9%增加到 3 4.9% ;在 48h时G1期增加到 76.2 % ,S期减少到 11.4% ;72h各期恢复到正常比例。对照组细胞转染前后形态变化不明显 ,RGS16蛋白表达阴性 ,细胞周期变化不明显。结论　RGS16能促进C6细胞周期的运行 【英文摘要】 Objective To study the effect of regulator of G protein signaling 16(RGS16) on the cell cycle of glioma C6 cell.Methods pCMV5 RGS16 was transfected into C6 cells by lipofectin.The morphological and adhesive changes of the cells were observed under an inverted microscope.Expression of RGS16 was examined by immunocytochemical method both before and after the transfection.Flow Cytometry was adopted to measure the fraction number changes of the cell cycle phase every 12 h.Results 24 hours after the transfec..

Promotion of glioma C6 cells proliferation by overexpressed RGS16

目的　探讨 G蛋白调节子 16 (RGS16 )对大鼠胶质瘤C6细胞的生物学特性的影响 .方法　利用脂质体介导法将RGS16基因导入 C6细胞中 ,在倒置显微镜下观察细胞形态变化和贴壁情况 ;3H- Td R法检测 C6细胞在转染不同梯度p CMV5 - RGS16和 p CMV 5质粒后的增殖情况 ;免疫细胞化学法检测转染前后 RGS16蛋白的表达情况 ;流式细胞仪检测转染 p CMV5 - RGS16和 p CMV5质粒 36 h后细胞周期变化和细胞是否有凋亡发生 .结果　转染 p CMV5 - RGS16质粒 2 4 h后 30 %细胞贴壁性降低 ,突起收缩 ,细胞变圆 ;RGS16蛋白表达阳性 ;3H- Td R法检测显示 C6细胞增殖速度与转染p CMV5 - RGS16的量呈正相关 ;细胞周期结果显示 G1期细胞百分数减少 10 % ,而 S期细胞百分数增多 14 % ;未发现RGS16与凋亡有直接关系 .结论　 RGS16可能促进 C6细胞的增殖 . 【英文摘要】 s: AIM To study the effect of RGS16 on the biological characteristics of glioma C6 cells. METHODS pCMV5 RGS16 was transfected into C6 cells by lipofectin. The morphological and adhesive changes of the cells were observed under an inverted microscope. Proliferation of C6 cells was measured by 3H thymidine ( 3H TdR) assay after gradient transfections of pCMV5 RGS16 and pCMV5. Expression of RGS16 was examined by immunocytochemical method both before and after the transfection. Flow cytometry ...高等学校骨干教师计划资

In-situ Photochemical Fluorimetry of Tyrosine and Its Determination

在氨性缓冲溶液中，酪氨酸经２７５ｎｍ紫外光照射后，产生荧光二聚体。据此，建立了现场光化学荧光法测定酪氨酸的新方法。线性范围为 ２．０ × １０－７～ ２．０ × １０－５ ｍｏｌ／Ｌ。检测限及精密度分别为 ３．２ × １０－９ｍｏｌ／Ｌ－１和 １．７％。方法简便、快捷，对其它氨基酸有较好的选择性，可用于啤酒及氨基酸注射液的测定。文章对光化学反应机理作了初步探讨。Tyrosine can produce fluorescent dimers under irradiation with 275nm UV-light in NH3-NH4CI buffer. Besed on this fact, a new photochemical fluorimetry in-situ was developed for determination of tyrosine. The linear relationship is over the range of 2. 0 x 10-7 - 2. 0 x 10-5 mol/L tyrosine. The detection limit is 3 .2 x 10-9 mol/L and relative standard deviation is 1 .7%. This method is simple and rapid, and has better selectivity for other amino-acids. It has been applied to the determination of tyrosine in beer and amino-acid injections. Mechanism of the photochemical reaction was discussed.福建省自然科学基金资助项目

A Study on Light as a Substitute for Glutaminic Acid Oxidase in the Enzymatic Reaction of Glutaminic Acid

在弱碱性 (p H=10 .6 )介质中 ,紫外光的照射引起谷氨酸的光化学反应 ,产生 H2 O2 和 NH3 ,与谷氨酸氧化酶存在下的酶促反应等效 .以萘斯勒试剂使产物中的 NH3 显色并用分光光度法测定 ,以及以氯化血红素 (hemin)作为过氧化物酶的替代物 ,催化产物中的 H2 O2 氧化、偶联对羟基苯乙酸(HPA)的荧光反应并进行荧光检测 .据此分别建立了间接测定谷氨酸的新方法 .In week alkaline medium(pH=10.6),photochemical reaction of glutaminic acid can occur under the irradiation of UV light and produces NH 3 and H 2O 2.This reaction is equal in value to enzymatic reaction of glutaminic acid in the presence of glutaminic acid oxidase.NH 3 produced by photochemical reaction is coloured by Nessler′s reagent and the absorbance is measured.However, using hemin as a substitute of peroxidase,H 2O 2 produced by photochemical reaction oxidazes p hydroxyphenyl acetic acid(HPA) and couses the coupled fluorescence reaction.Accodingly,the spectrophotometry and fluorimetry have been developed for indirect determination of glutaminic acid,respectively.福建省自然科学基金资助项目!(B 96 0 0 3

Light as a Substitute for Glutaminic Acid Oxidase and Fluorimentric Determination of Glutaminic acid

在弱碱性介质中，紫外光的照射引起谷氨酸的光化学反应，产生Ｈ２Ｏ２和ＮＨ３，与谷氨酸氧化酶存在下的酶促反应等效。利用氯化血红素（ｈｅｍｉｎ）作为过氧化酶的替代物，催化Ｈ２Ｏ２与对羟基苯乙酸（ＨＰＡ）的偶合荧光反应，建立了间接荧光法测定谷氨酸的方法。Under the irradiation of UV-light, photochemical reaction of glutaminic acid occur and produces H2O2 and HN3 in weak alkaline medium with acetone. This reaction is equal in value for enzymatic reaction of glutaminic acid in the presence of glutaminic acid oxidase. Then, using hemin as catalyst, H2O2 produced by photochemical reaction oxidase p-hydroxyphenyl acetic acid (HPA) and causes fluorescence reaction coupled of it. Based on this result, a new fluorimetric method was developed for indirect determination of glutaminic acid.福建省自然科学基金资助项

In-situ photochemical fluorimetry of tyrosine and its determination

Tyrosine can produce fluorescent dimers under irradiation with 275nm UV-light in NH3-NH4Cl buffer. Besed on this fact, a new photochemical fluorimetry in-situ was developed for determination of tyrosine. The linear relationship is over the range of 2.0 x 10(-7) similar to 2.0 x 10(-5) mol/L tyrosine. The detection limit is 3.2 x 10(-9) mol/L and relative standard deviation is 1.7%. This method is simple and rapid, and has better selectivity for other amino-acids. It has been applied to the determination of tyrosine in beer and amino-acid injections. Mechanism of the photochemical reaction was discussed

Light as a substitute for glutaminic acid oxidase and fluorimetric determination of glutaminic acid

Under the irradiation of UV-light, photochemical reaction of glutaminic acid occur and produces H2O2 and NH3 in weak alkaline medium with acetone. This reaction is equal in value for enzymatic reaction of glutaminic acid in the presence of glutaminic acid oxidase. Then, using hemin as catalyst, H2O2 produced by photochemical reaction oxidazes p-hydroxyphenyl acetic acid (HPA) and causes fluorescence reaction coupled of it. Based on this result, a new fluorimetric method was developed for indirect determination of glutaminic acid