一氧化氮合酶在糖尿病大鼠视网膜中表达模式的研究

目的 研究一氧化氮合酶(NOS)在8周糖尿病大鼠视网膜的表达,探讨其与一氧化氮在糖尿病视网膜病变(DR)中可能的分子作用机制.方法 采用限制片段差异显示PCR(RFDD-PCR)技术建立正常和8周糖尿病大鼠视网膜基因表达谱,经生物信息学分析两者差异,初步确定NOS三种亚型-eNOS、nNOS和iNOS为DR相关基因,并以半定量RT-PCR和免疫组化方法进行验证.结果 RFDD-PCR结果显示,糖尿病组eNOS和nNOS表达下调,iNOS表达上调.RT-PCR结果显示,糖尿病组eNOS和nNOS表达比正常组明显降低(eNOS:0.23±0.03,0.32±0.03,P＜0.05;0.25±0.02,0.36±0.02,P＜0.05),iNOS比正常组明显增高(0.27±0.02,0.20±0.03,P＜0.05).免疫组化结果显示,正常组eNOS、nNOS和iNOS阳性细胞均见于内核层(INL)和节细胞层(GCL),eNOS阳性细胞也分布于血管内皮层;糖尿病组eNOS和nNOS阳性细胞较正常组明显减少(eNOS:14.33±3.19,22.13±3.60,P＜0.05;nNOS:21.87±3.62,34.40±7.09,P＜0.05),iNOS较正常组明显增多(17.60±2.58,11.73±2.70,P＜0.05).结论 eNOS、nNOS和iNOS表达变化与DR发生发展有关.     

L-精氨酸对大鼠尾核一氧化氮合酶基因表达的影响

目的 研究尾核中一氧化氮在痛觉调制中的作用机制.方法 大鼠尾核内微量注射L-精氨酸、Nω-硝基-L-精氨酸甲酯、生理盐水,应用逆转录-聚合酶链反应测定4,8,12,24和48 h大鼠尾核nNOS mRNA表达的变化;新生Wistar大鼠尾核神经元原代培养液中加入L-精氨酸、Nω-硝基-L-精氨酸甲酯、生理盐水,观察给药12 h尾核nNOS mRNA表达的变化.结果 大鼠尾核给药24 h内L-精氨酸组nNOS mRNA表达较正常增强(P＜0.05),而Nω-硝基-L-精氨酸甲酯组nNOS mRNA表达较正常减弱(P＜0.05),48 h恢复正常.体外尾核原代神经元培养,nNOS mRNA表达的变化趋势和在体实验相一致.结论 中枢神经系统尤其是尾核中NOS活性是一氧化氮作为神经递质或调质参与中枢痛觉调制的关键因素之一.     

当归芍药散对阿霉素肾病综合征大鼠一氧化氮及其合酶表达影响

目的 :尾静脉注射阿霉素法建立肾病综合征(Nephrotic Syndrome,NS)大鼠,探讨当归芍药散对肾病综合征大鼠一氧化氮合酶表达的影响及其机制研究。方法 :将雄性SD大鼠复制为NS模型,随机分为阳性对照组、模型组和当归芍药散组,另设正常对照组。灌胃治疗后,HE染色观察肾脏组织病理变化;BCA法检测24 h尿蛋白量;分光光度法测定尿液NO含量;WB法检测肾组织内皮型一氧化氮合酶(endothelial NOS,eNOS)、诱导型一氧化氮合酶(inducibleNOS,iNOS)和神经型一氧化氮合酶(neural NOS,nNOS)的表达。结果 :模型组与正常组比较,尿蛋白量显著增加(P0.01),肾脏病理损伤严重;各治疗组与模型组比较尿蛋白含量下降明显,差异具有显著性(P0.01);模型组与正常组比较大鼠尿液NO含量降低显著(P0.01),当归芍药散组与模型组比较NO含量显著增加(P0.01);与正常组比模型组iNOS、nNOS和eNOS蛋白表达均降低,且差异具有显著意义(P0.01);与模型组比较各给药组能显著增加iNOS、nNOS和eNOS蛋白表达,差异具有极显著意义(P0.01)。结论 :当归芍药散可以提高肾病综合征大鼠NOS的表达,增加大鼠体内NO含量,减少尿蛋白含量,延缓肾病综合征病理进程。     

L-硝基精氨酸对大鼠局灶性脑缺血损伤的保护作用及其机制

目的:研究非选择性一氧化氮合酶(NOS)抑制剂L-硝基精氨酸(L-NA)对局灶性脑缺血大鼠脑组织的保护作用及其机制。方法:线栓法复制大鼠大脑中动脉脑缺血模型;采用RT-PCR法检测大鼠缺血0,2,6,12,24 h脑组织中[内皮型NOS(eNOS)、神经型NOS(nNOS)及诱导型NOS(iNOS)]基因表达的变化;TTC染色法测定大鼠脑梗死体积,HPLC法测定纹状体、海马、皮层中氨基酸含量。结果:正常对照组可见eNOS和nNOS表达,eNOS于脑缺血后2 h达到高峰,nNOS于6 h达到高峰;正常对照组未见iNOS表达,但在脑缺血后开始表达,缺血后12 h达到高峰;缺血后12 h给予L-NA治疗3 dL-NA组中梗死体积/全脑体积(Ⅳ%)显著低于相应的缺血组;缺血组天门冬氨酸、谷氨酸、甘氨酸、GABA的含量明显高于假手术组,缺血12 h治疗3 d时,L-NA组中纹状体、海马、皮层中天门冬氨酸、谷氨酸的含量显著低于缺血组,GABA的含量显著高于缺血组,海马中甘氨酸含量显著高于缺血组。结论:缺血后期应用L-NA对脑缺血有治疗作用,可能与兴奋性氨基酸合成、释放相对减少,抑制性氨基酸合成、释放相对增加有关。     

安氟醚对大鼠耳蜗一氧化氮合酶表达的影响

目的:观察安氟醚对大鼠耳蜗一氧化氮合酶(NOS)表达的影响,探讨其对耳蜗影响的可能机制。方法:30只Wistar大鼠随机分为3组(n=10),对照组(C组)持续吸入纯氧;低浓度安氟醚组(E1组)持续吸入1.5%安氟醚+纯氧;高浓度安氟醚(E2组)持续吸入3.0%安氟醚+纯氧,30min后处死大鼠,免疫组织化学方法检测耳蜗螺旋器、血管纹和螺旋神经节诱生型NOS(iNOS)、内皮型NOS(eNOS)和神经元型NOS(nNOS)的表达水平。结果:与C组比较,E1组耳蜗螺旋器、螺旋神经节iNOS、eNOS和nNOS以及血管纹eNOS表达均下调,E2组耳蜗螺旋器、血管纹和螺旋神经节iNOS、eNOS和nNOS表达下调(P<0.05或0.01);E2组耳蜗螺旋神经节eNOS和nNOS表达组较E下调明显(P<0.05)。结论:安氟醚可浓度依赖性地下调大鼠耳蜗NOS表达,从而影响耳蜗功能。     

内皮型一氧化氮合酶基因多态性与脑梗死的研究

一氧化氮合酶(Nitric Oxide Synthase，NOS)的一个重要指征是不仅合成一氧化氮(Nitric Oxide，NO)，也能产生超氧离子。目前已确定的NOS的亚型有三种，根据不同的基因编码分别称为神经元型NOS(neuronal NOS，nNOS)、诱导型NOS(inducibl eNOS，iNOS)、内皮型NOS(endothel ialNOS，eNOS)。三种NOS亚型在血管内皮细胞中均可表达，但在生理情况下，血管内皮细胞中的eNOS合成和释放NO无疑是血管张力的主要调节因子。而nNOS与iNOS产生的NO则主要产生氧化作用，具神经毒性作用。     

甲状腺激素对新生鼠脑NOS基因表达的调节

目的"观察不同日龄新生鼠脑一氧化氮(NO)含量,一氧化氮合酶(NOS)活性的变化及甲状腺激素对nNOS基因表达的调节. 方法"采用丙基硫氧嘧啶(PTU)灌注孕母鼠造成甲减模型.采用NO、NOS的生化测定法以及半定量逆转录聚合酶链反应(RT-PCR). 结果"生化测定法显示,不同年龄段大鼠随着年龄的增加NO含量上升(P＜0.01),NOS的活性上升(P＜0.01);同一年龄段仔鼠,甲减组较正常组NO含量下降(P＜0.01),NOS活性下降(P＜0.01);RT-PCR测定显示,幼鼠nNOS mRNA水平随着年龄的增加而上升;同一年龄组,甲减组较正常组nNOS mRNA表达下降. 结论"提示甲状腺激素对nNOS mRNA表达有上调作用;NO信号系统可能参与鼠脑发育阶段甲状腺激素缺乏所致的脑损害过程.     

雄激素致无排卵大鼠卵巢nNOS、eNOS、ET-1的表达

目的 观察雄激素致无排卵大鼠(ASR)血浆及卵巢血管舒张因子一氧化氮(NO)、一氧化氮合酶(nNOS、eNOS)、环磷酸鸟苷(cGMP)、血管收缩因子内皮素1(ET-1)、血管紧张素Ⅱ(ATⅡ)的变化,探讨卵巢血管舒-缩因子在无排卵发病机制中的作用.方法 建立雄激素致无排卵大鼠模型,分为模型组和正常组.采用放射免疫法测定模型组、正常组大鼠血浆NO、cGMP、ET-1、ATⅡ以及血清雌二醇(E2)、孕酮(P)、睾酮(T)含量;采用免疫组化法检测大鼠卵巢nNOS、eNOS、ET-1表达;采用HE染色观察大鼠卵巢组织形态学变化.结果 与正常组比较,模型组大鼠血浆NO、cGMP含量降低(P<0.01),血浆ET-1、ATⅡ含量升高(P<0.01).血清E2、T含量及E2/P显著升高(P<0.01),血清P含量无统计学差异(P>0.05);与正常组比较,模型组大鼠卵巢颗粒细胞以及髓质nNOS、eNOS表达减弱(P<0.01),ET-1表达增强(P<0.01);正常组大鼠卵巢皮质可见发育期各级卵泡及黄体;模型组大鼠卵巢皮质卵泡囊状扩张,颗粒细胞层极薄.结论 雄激素致无排卵大鼠血浆及卵巢血管舒张因子降低、收缩因子增高,可能使卵巢血液供应障碍、颗粒细胞凋亡,导致卵泡在排卵前闭锁,无排卵的发生.     

内皮型一氧化氮合酶基因在犬内皮细胞的转染和表达

目的探讨转染内皮型一氧化氮合酶(eNOS)基因后内皮细胞(EC)的功能变化.方法采用脂质体法转染eNOS基因于实验犬EC;RT-PCR和免疫组化法检测转染效果;分别采用比色法和酶联免疫法检测细胞培养液一氧化氮合酶(NOS)、一氧化氮(NO)和vWF的浓度;并观察转染细胞生长增殖情况.结果 RT-PCR产物电泳和测序及免疫组化法检测证实转染效果满意;转染后EC培养液NOS和NO浓度在不同时间明显升高(120 h分别为33.53和32.99),与正常组对比差异显著(P<0.05).转染后细胞生长增殖和vWF含量无显著差异.结论通过脂质体法成功地将eNOS基因转染于实验犬EC;转染后eNOS基因在mRNA和蛋白质水平均高效表达;内皮细胞eNOS活性显著增强;转染后细胞生物学功能稳定.     

地塞米松对内毒素休克新生大鼠脑一氧化氮合成酶基因表达的调节

目的：探讨新生大鼠内毒素休克脑损伤时脑一氧化氮合成酶(NO6)三种亚型基因表达的变化及地塞米松(DEX)对其的调控作用。方法：在新生大鼠内毒素休克动物模型基础上，采用逆转录PCR及PCR技术，对脑组织中三型NOS mRNA及caspase—3 mRNA的表达进行半定量分析。结果：正常新生大鼠脑iNOS及eNOS mRNA无明显表达，nNOS mRNA、caspase—3 mRNA有一定程度表达。内毒素脂多糖(LPS)腹腔注射后2h，三种亚型NOS mRNA开始表达，于LPS6h达高峰，并持续至24h。caspase—3 mRNA于LPS腹腔注射后2h后表达逐渐增加，24h达高峰。DEX可抑制nNOS、iNOS及caspase—3 mRNA的表达，且以用药后2h最为明显，并持续至用药后24h。结论：内毒素休克脑损伤时，各型NOS均有表达，NO的产生是内毒素休克脑损伤时重要的病理生理机制之一。DEX通过抑制NOS、caspase—3 mRNA的表达部分实现其神经保护作用．     

Acupuncture Treatment of Vomiting

CASE HISTORY A female patient, 46 years old, head of the foreign affairs department of a certain university in Beijing, paid her first visit on October 9, 2006, with the chief complaint of vomiting for one month. She got vomiting after meals in early September. Before that, she had discomfortable sensation in the stomach due to angry with others, but she didn't pay much attention. Later, it developed into vomiting after eating. After the vomiting, the discomfort would be relieved, but with slight hypodynamia. She was once diagnosed as having 'neurogenic vomiting'. Having taken some western and Chinese drugs, the above symptoms were a little bit improved, but she would have nausea upon eating and with regurgitation. Because of the fear for vomiting, she did not dare to have food intake, with body weight reduction of 6 kilos in one month.     

Qiu Baoguo''''s Experience in Treating Consumptive Syndromes after Radio-Chemotherapies

Radiotherapy and chemotherapy are the important modern medical therapies for malignant tumors,yet they can also bring about serious local and systemic toxic side reactions so to decrease the patient;'s life quality,manifested by a series of consumptive symptoms.Having engaged in the combined work of Chinese and western medicine for nearly 50 years,the research fellow Qiu Baoguo in Henan Provincial Academy of TCM has developed his unique views on the TCM study of consumptive syndromes.The author of this essay had once the fortune tO follow Dr.Qiu in clinic,and specially would like to introduce in the following Dr.Qiu's experience in treating consumptive syndromes after radio-chemotherapies for patients with malignant tumor.     

Insomnia Due to Deficiency of Both the Heart and Spleen Treated by Acupuncture-Moxibustion and Chinese Tuina

OBJECTIVE: To observe therapeutic effects of the comprehensive therapy of acupuncture-moxibustion and Chinese Tuina for treatment of insomnia due to deficiency of both the heart and spleen. METHODS: 92 cases were divided randomly into the treatment group (treated by acupuncture-moxibustion and Chinese Tuina) and the control group (treated by acupuncture-moxibustion). RESULTS: The therapeutic effect of the treatment group was obviously superior to that of the control group (the CHI2 test showed P < 0.01). CONCLUSIONS: The comprehensive therapy of acupuncture-moxibustion and Chinese Tuina can give marked therapeutic effects for treatment of insomnia due to deficiency of both the heart and spleen.     

Analysis of CK-19,PDX-1,Nestin,Ngn3 in different donor islet purity in rats

Objective: To investigate if there are the CK-19, PDX-1, Nestin, Ngn3 positive cells in the donor islets of different purity in rats. Methods: Thirty male adult SD rats were randomly divided into 3 groups. Islets were isolated using digestion by ductal injection of collagenase. Group Ⅰ (n=10): Separating cell preparations were not purified, Group Ⅱ(n=10): Islet sediment was purified with 25% Ficoll400 ,Group Ⅲ (n=10): Islet sediment was purified with 25% and 11% Ficoll-400. The levels of protein of CK-19, PDX-1, Nestin and Ngn3 were detected by immunohistochemistry and the mRNA of CK-19, PDX-1, Nestin, Ngn3 was amplified by RT-PCR. Results: After two different purification methods applied, three islet preparations of different purities were obtained. The difference of islet purity was significant among various groups (P＜0.05). Compared with group Ⅱ and group Ⅲ,the protein and mRNA of CK-19, PDX-1, Nestin,Ngn3 were both higher in group Ⅰ; group Ⅲ was poorly expressed. Conclusions: The three different islet purity donor islet have different CK-19, PDX-1, Nestin, Ngn3 positive cells within them, indicating that there are some islet stem cells in the purified donor islet.     

A preliminary screening study on the associated proteins in human psoriasis vulgaris by serum proteomics technologies

Objective：To investigate the optimum screening conditions of associated proteins in human psoriasis vulgaris by serum proteomics technique, and to screen the different expression proteins related with psoriasis vulgaris. Methods：Serum samples of peripheral blood were collected from newly diagnosed psoriasis vulgaris patients in the clinic, and 20 matched healthy persons. Serum albumin IgG was removed by filtering with ProteoExtract Albumin/IgG. After comparative proteomics analysis the different protein spots were identified using 2-DE and MS. Results ：Electrophoresis figures with high resolution and reproducibility were obtained. Three different expression proteins were found only in the serum from psoriasis vulgaris patients, while nine other different proteins expressing from healthy volunteers. Conclusion：The protein expression was different in the serum between the psoriasis vulgaris patients and healthy volunteers. It was hoped that we could find the biomarkers related to psoriasis vulgaris by using proteomics.     

A CASE OF HUGE GASTRIC DISOPYROBEZOAR

PYTOBEZOARS are common bezoars in gastrontestinal tract, commonly seen in stomach and mall intestine,1but huge disopyrobezoars are     

Acupuncture Treatment of Chest Bi Syndrome

CASE HISTORY A male,a retired officer of 61 years old,paid his first visit on May 12,2006.The patient stated that 2 weeks before he suddenly got paroxysmal precordial choking pain,with irregular attacks.Each attack would last several seconds to several minutes.The attack was accompanied with heavy oppressed sensation in the precordial region,palpitation,lassitude,and in severe cases,sweating,which was once diagnosed by a hospital as'coronary heart disease'.Recalling the case history,the patient said that several days before he got angry with others because of some affairs in work,and he was not so used to the life pattern since he had just retired from his busy working post.At usual times,he was quick tempered and peevish.In recent days,he had poor appetite,and loose stool.     

Instructions to Contributors

The English edition of Chinese Journal of Integrative Medicine is edited and published by the Chinese Journal of Integrated Traditional and Western Medicine Press.Original scientific papers of advanced clinical and experimental medicine in the field of complementary/alternative medicine or integrative medicine are expected.