神经组织蛋白质类

摘要： 目的:分析神经元特异性烯醇化酶(neuron specific enolase,NSE)、S100B、胶质纤维酸性蛋白(glial fibrillary acidic protein,GFAP)在结核性脑膜炎(tuberculous meningitis,TBM)患儿血清及脑脊液中表达的动态变化特征及其对患儿预后预测价值。方法:采用前瞻性研究方法,连续纳入2018年1月1日至2021年12月31日于南华大学附属长沙中心医院儿童结核科收治的72例TBM患儿作为TBM组。搜集同期住院,诊断为肺结核同时排除TBM的患儿20例作为肺结核组。TBM组患儿治疗6个月后根据预后情况,分为完全康复组(47例)和预后不良组(25例)。采用酶联免疫吸附试验法测定TBM组入院48 h内及治疗后(1、2、3周)和肺结核组入院48 h内的血清和脑脊液NSE、S100B、GFAP水平,并进行比较。采用受试者工作特征(receiver operating characteristic,ROC)曲线及曲线下面积(area under the curve,AUC)分析TBM患儿入院时脑脊液NSE、S100B、GFAP水平,预测其不良预后的阈值、敏感度及特异度。结果:入院时血清及脑脊液NSE、S100B、GFAP水平[中位数(四分位数)],TBM组分别为15.12(3.22,26.90)μg/L、1.11(0.40,3.20)μg/L、15.34(3.44,45.82)μg/L和37.90(6.50,142.70)μg/L、2.31(1.02,10.20)μg/L、65.31(10.87,252.60)μg/L,均明显高于肺结核组[分别为7.03(2.48,13.23)μg/L、0.25(0.12,0.36)μg/L、10.38(2.41,19.00)μg/L和7.56(2.12,12.79)μg/L、0.35(0.05,0.51)μg/L、7.86(2.41,13.80)μg/L],差异均有统计学意义(血清水平:Z值分别为-5.064、-6.817、-2.693,P值分别为0.000、0.000、0.007;脑脊液水平:Z值分别为-6.465、-6.816、-6.778,P值均为0.000);预后不良组脑脊液NSE、S100B、GFAP水平分别为60.16(24.90,142.70)μg/L、2.59(1.32,10.20)μg/L、118.74(58.83,252.60)μg/L,明显高于完全康复组[分别为29.37(6.50,68.82)μg/L、1.97(1.02,6.10)μg/L、45.39(10.87,84.93)μg/L],差异均有统计学意义(Z值分别为-4.855、-3.212、-6.334,P值分别为0.000、0.001、0.000)。治疗后1、2、3周,预后不良组脑脊液NSE分别为49.58(15.38,87.56)μg/L、41.53(9.60,82.00)μg/L、25.97(5.56,58.49)μg/L,S100B分别为10.15(3.63,15.72)μg/L、1.60(0.41,3.28)μg/L、0.75(0.41,1.60)μg/L,GFAP分别为99.75(65.79,180.84)μg/L、63.94(13.65,120.59)μg/L、38.03(10.87,85.40)μg/L,均明显高于完全康复组[NSE分别为18.49(4.87,36.12)μg/L、14.51(4.87,35.70)μg/L、8.53(2.12,21.70)μg/L;S100B分别为5.34(2.19,10.08)μg/L、0.66(0.19,1.56)μg/L、0.40(0.11,0.74)μg/L;GFAP分别为45.39(10.87,84.93)μg/L、17.77(5.66,38.15)μg/L、12.82(5.04,26.90)μg/L,差异均有统计学意义(NSE:Z值分别为-2.496、-3.815、-4.041,P值分别为0.013、0.000、0.000;S100B:Z值分别为-3.331、-4.745、-1.207,P值分别为0.047、0.000、0.036;GFAP:Z值分别为-4.940、-2.337、-3.745,P值分别为0.000、0.016、0.012)。ROC曲线分析,获得最大约登指数下入院时TBM患儿脑脊液NSE、S100B、GFAP水平对预后不良的预测阈值分别为51.92μg/L、2.75μg/L、77.54μg/L。结论:TBM患儿血清及脑脊液NSE、S100B、GFAP水平明显增高,经治疗后血清NSE、S100B、GFAP水平快速降至正常,而脑脊液NSE、S100B、GFAP水平下降缓慢。TBM患儿入院时脑脊液NSE≥51.92μg/L、S100B≥2.75μg/L或GFAP≥77.54μg/L,且经治疗后脑脊液NSE、S100B、GFAP下降缓慢,提示预后不良的可能。

摘要： 目的 研究富含亮氨酸重复序列的神经元3(LRRN3)在非小细胞肺癌增殖中的作用及其表达调控的可能机制.方法 运用实时定量PCR、免疫组织化学和生物信息检索检测LRRN3在非小细胞肺癌中的表达差异;利用慢病毒过表达技术,建立稳定过表达LRRN3的肺癌细胞系A549-LRRN3;MTT实验检测LRRN3对非小细胞肺癌增殖能力的影响;生物信息学检索LRRN3启动区甲基化的改变,且通过甲基化转移酶抑制剂处理肺癌细胞,检测甲基化对LRRN3表达调控的影响;最后生物信息学检索分析LRRN3与肺癌预后的相关性.结果 qPCR检测发现LRRN3在非小细胞肺癌(12例)临床组织标本中的mRNA表达水平明显低于癌旁正常组织(12例)(P=0.0014);免疫组化结果显示LRRN3在非小细胞肺腺癌中的蛋白表达水平低于正常组织(P=0.001),同时在非小细胞肺鳞癌中的表达也低于正常组织(P=0.003);过表达LRRN3可抑制肿瘤细胞的增殖(P<0.01);并且LRRN3的启动子区存在高甲基化修饰抑制其转录表达,LRRN3表达与肺腺癌的生存预后正相关(P=5.2e-09;HR=0.48).结论 LRRN3的启动区高甲基化抑制LRRN3转录表达,进而促进肺癌细胞的增殖.%Objective To study the role of leucine rich repeat neuronal 3 (LRRN3) in the prolif-eration of non-small cell lung cancer and its possible mechanism of expression regulation. Methods The expression of LRRN3 in non-small cell lung cancer was detected by real-time quantitative polymerase chain reaction ( qPCR) , immunohistochemistry and bioinformatics retrieval;A lung cancer cell line A549-LRRN3 with stable over-expression of LRRN3 was established by lentivirus over-expression technology;The effect of LRRN3 on the proliferation of non-small cell lung cancer was detected by methyl thiazolyl tetrazolium ( MTT) assay;Bioinformatics search for changes in methylation of LRRN3 promoter region and treatment of lung cancer cells by methyltransferase inhibitors to detect the effect of methylation on the regulation of LR-RN3 expression; Finally, bioinformatics search analyzes the correlation between LRRN3 and lung cancer prognosis. Results The mRNA expression of LRRN3 in clinical tissues of non-small cell lung cancer (n=12) was significantly lower than that of adjacent normal tissues (n=12) (P=0. 0014). The results of immunohistochemistry showed that the protein expression level of LRRN3 in non-small cell lung adenocar-cinoma was lower than that in normal tissues (P=0. 001), and the expression in non-small cell lung squa-mous cell carcinoma was also lower than that in normal tissues (P=0. 003). Overexpression of LRRN3 in-hibited the proliferation of tumor cells (P<0. 01), and the hypermethylation of LRRN3 in the promoter re-gion inhibited its transcriptional expression. LRRN3 was positively correlated with the survival prognosis of lung adenocarcinoma (P=5. 2e-09;HR=0. 48). Conclusions Hypermethylation in the promoter region of LRRN3 inhibits its transcriptional expression, thereby promoting the proliferation of lung cancer cells.

摘要： Objective To investigate the protection effects of the activation of NMDAR1(NMDA receptor 1)/ERK1/2 signal pathway on visual cortex nerve cells induced by levodopa in amblyopia rats.Methods SD rats of SPF grade,were randomly divided into for groups of 9,group A.Control,B.MD group,C.L-levodopa(20 mg· kg-1)+MD group,D.H-levodopa (80 mg· kg-1)+MD group,E.H-levodopa+MD+MK801 group,F.H-levodopa+MD+PD98059 group,G.MD+MK801 group,H.MD+PD98059 group,Ⅰ.MD+DMSO group.Amblyopia rats were made by suture of the right eye.Levodopa was used to treatment amblyopia by gavage,and intervened by intracerebroventricular injection of MK801 and PD98059 respectively.The expression of NR1,p-ERK1/2,ERK1/2,NGF and c-FOS were detected by Western blotting.Nissl staining was used to detect morphological changes of neurons.Neuronal apoptosis was detected by TUNEL method,and detected the expression of Caspase-3,NGF and c-FOS by immunohistochemical staining.One/Two way Chi-square analysis was used for data analysis.LSD-t test was used as comparison between every two groups.Results The morphology of Nissl bodies in neurons was complete and clear in A group,and the size of Nissl bodies got smaller,and caused karyopyknosis and loss of neurons in visual cortex of B group.Compared with A group,the apoptosis of visual cortical neurons(23.09±2.00 vs.2.20±0.35,t=12.120,P=-0.000) and the number of Caspase-3 postive cells (22.70± 1.50 vs.3.30 ± 0.54,t=12.120,P=0.000)were significantly increased,the expression of NGF(0.31 ±0.04 vs.0.74±0.09,t=7.674,P=0.000) and c-FOS(0.25± 0.03 vs.0.57±0.07,t=5.919,P=0.000) and the rats of NGF(8.30±0.82 vs.35.18±2.01,t=12.37,P=0.0000) and c-FOS (10.84± 1.02 vs.35.68±2.55,t=9.056,P=0.0001) postive cell were decreased significantly in B group.After treatment with levodopa,the morphology of neurons recovered,the apoptosis of visual cortical neurons relieved,the expression of NR1(0.75±0.09 vs.0.40±0.05,t=8.528,P=0.001) and p-ERK1/2(2.13± 0.26 vs.0.68±0.17,t=3.488,P=-0.008) were increased significantly,and the rats of NGF (18.07±0.87 vs.8.30± 0.82,t=8.18,P=0.0000) and c-FOS (19.78 ±0.91 vs.10.84± 1.02,t=6.543,P=0.0001) postive cells were significantly increased.MK-801 or PD98059 intervention could effectively attenuate the effect of levodopa.It could effectively down-regulated the expression of NR 1 (0.53 ±0.06 vs.0.95 ± 0.12,t=5.647,P=0.005) and p-ERK1/2(1.52±0.18 vs.2.58±0.30,t=3.091,P=0.013) interference with MK801 or PD98059 in MD rats.MK-801 or PD98059 intervention further promote the Nissl body volume reduced,neurons karyopyknosis,the apoptosis of visual cortical neurons and Caspase-3 expression,and restrain the expression of NGF and c-FOS.Conclusion Levodopa played a protective role in visual cortex nerve cells of amblyopia rats at least partially through activation of NMDA-ERK1/2 signal pathway.%目的 探讨左旋多巴活化NMDA受体1(NMDAR1)/ERK1/2(丝裂原活化蛋白激酶)信号对单眼剥夺弱视大鼠视皮质神经细胞的保护作用.方法 实验研究.54只SD大鼠根据随机数字表分为9组:正常对照组、弱视模型(MD)组、低剂量左旋多巴治疗组、高剂量左旋多巴治疗(H-左旋多巴+MD)组、高剂量左旋多巴+MK801组、高剂量左旋多巴+PD98059组、MK801组、PD98059组、二甲基亚砜溶剂对照组,每组6只.通过右眼缝合制备弱视大鼠模型,左旋多巴灌胃治疗,MK801、PD98059通过侧脑室注射,免疫印迹法检测N-甲基-D-天冬氨酸受体(NR1)、磷酸化丝裂原活化蛋白激酶(p-ERKI/2)、丝裂原活化蛋白激酶(ERK1/2)、神经生长因子(NGF)、即刻早期基因c-FOS的表达,Nissl染色检测神经元细胞形态学变化,核苷酸末端转移酶介导的dUTP缺口标记(TUNEL)法检测神经元细胞凋亡,免疫组织化学法检测神经元细胞半胱氨酸天冬氨酸蛋白酶-3(caspase-3)、NGF和c-FOS的表达.采用单因素或双因素方差分析联合LSD-t检验进行组间比较.结果 正常对照组小鼠神经元尼氏小体完整,形态清晰;MD组视皮质区神经元尼氏小体体积缩小,神经元丢失和核固缩.与对照组相比,MD组神经细胞凋亡显著增多(23.09±2.00、2.20±0.35,t=12.120,P=0.000),Caspase-3阳性细胞增多(22.70±1.50、3.30±0.54,t=12.120,P=0.000),NGF (0.31 ±0.04、0.74±0.09,t=7.674,P=0.000)和c-FOS (0.25±0.03、0.57±0.07,t=5.919,P=0.000)表达降低,NGF(8.30±0.82、35.18±2.01,t=12.370,P=0.0000)和c-FOS (10.84±1.02、35.68±2.55,t=9.056,P=0.0001)阳性细胞数减少,视皮质区NR1 (0.40±0.05、0.55±0.07,t=4.533,P=0.001)、p-ERK1/2(0.68±0.17、0.88±0.11,t=0.920,P=0.142)的表达降低;左旋多巴治疗后,神经细胞形态恢复正常,尼氏小体染色清晰;有效缓解神经细胞凋亡(13.07±1.47、23.09±2.00,t=8.698,P=0.000),减少Caspase-3阳性细胞率(17.05±1.11、22.70±1.50,t=3.019,P=0.015);NR1 (0.75±0.09、0.40±0.05,t=8.528,P=0.001)、p-ERK1/2(2.13±0.26、0.68±0.17,t=3.488,P=0.008)的表达显著升高;NGF(18.07±0.87、8.30±0.82,t=8.18,P=0.0000)和c-FOS(19.78±0.91、10.84±1.02,t=6.543,P=0.0001)阳性细胞率增加.MK-801或PD98059干预处理后,有效拮抗左旋多巴治疗作用.MK801、PD98059处理下调NR1(0.53±0.06、0.95±-0.12,t=5.647,P=0.005)及下游p-ERK1/2 (1.52±0.18,2.58±0.30,t=3.091,P=0.013)的表达,并进一步促进MD小鼠尼氏小体体积缩小、细胞核固缩、神经元凋亡、Caspase-3表达,并抑制NGF和c-FOS的表达;二甲基亚砜溶剂未见明显作用.结论 左旋多巴通过活化NMDA-ERK1/2 MAPK信号对大鼠视皮质神经元细胞的保护作用.

摘要： ObjectiveTo determine the optimal timing of antenatal taurine supplementation to improve neuron and neural stem cell proliferation in fetal rats with intrauterine growth restriction.Methods Twenty-five pregnant SD rats were randomly divided into five groups (five rats in each group): group A was the control group, group B to E were the fetal growth restriction (FGR) model groups with low-protein diet during the experiment, group C, D, and E were supplemented with taurine [300 mg/(kg·d)] at day 9, 11 and 15, respectively. The birth weight of newborn rats was measured after natural delivery. The rats with body weight two standard deviations lower than the average weight in group A were diagnosed as FGR. There were five litters of newborn rats in each group, and two were randomly selected from each litter, resulting in ten newborn rats in each group. Proliferating cell nuclear antigen (PCNA) and fatty acid binding protein 7 (FABP7) positive cell expression in newborn rat brain tissues were detected by immunohistochemistry. Single factor analysis of variance, LSD tests were used for statistical analysis.ResultsThe average birth weight of newborn rats in group A, B, C, D and E were (6.61±0.45), (4.65±0.23), (5.37±0.17), (5.74±0.21), and (5.00±0.24) g, respectively. Average birth weight was lower in group B than in group A (t=2.447), higher in group D and E than in group B (t=2.306 and 2.306), higher in group D than in group C and E (t=2.306 and 2.306), and the differences were statistically significant (allP0.05). The IOD in group D was higher than that in group E, and the difference was statistically significant (t=4.182,P<0.05).ConclusionsAntenatal taurine supplementation can promote neuron and neural stem cell proliferation in rats with FGR. The effect is most obvious on the 11th day of pregnancy, and may lead to the promotion of brain development.%目的：探讨孕鼠补充牛磺酸促进胎儿生长受限（fetal growth restriction，FGR）胎鼠神经元及神经干细胞增殖的最佳时机。方法25只Sprague-Dawley孕鼠随机分为5组（每组5只），A组为对照组，B~E组均为FGR组采用全程低蛋白饮食法建立FGR胎鼠模型，且C~E组分别于妊娠第9、11、15天补充牛磺酸300 mg/（kg·d）。孕鼠自然分娩后，称量新生鼠出生体重，低于对照组新生鼠平均出生体重2个标准差，判定为FGR。每组5窝新生鼠，每窝随机抽取2只，每组10只，采用免疫组织化学方法检测新生鼠脑组织增殖细胞核抗原（proliferating cell nuclear antigen，PCNA）、脂肪酸结合蛋白7（fatty acid binding protein 7，FABP7）阳性表达情况。统计学方法采用单因素方差分析、LSD检验。结果 A、B、C、D、E组新生鼠平均出生体重分别为（6.61±0.45）、（4.65±0.23）、（5.37±0.17）、（5.74±0.21）和（5.00±0.24） g，B组低于A组（t=2.447），D和E组均高于B组（t值分别为2.306和2.306），D组高于C和E组（t值分别为2.306和2.306），差异均有统计学意义（P值均＜0.05）。新生鼠脑组织PCNA阳性表达主要位于细胞核内；A、B、C、D、E组每高倍镜视野（×400）PCNA阳性细胞个数分别为（31.03±5.38）、（46.49±4.38）、（59.65±5.37）、（67.76±5.84）和（53.53±6.94）个，B组多于A组（t=2.110），C、D和E组PCNA均多于B组（t值分别为2.110、2.110和2.131），D组多于C和E组（t值分别为2.101和2.110），差异均有统计学意义（P值均＜0.05）。新生鼠脑组织FABP7阳性表达主要位于细胞质中，A、B、C、D、E组的积分吸光度值分别为451733.1±141452.3、207232.2±60525.2、333766.6±68412.1、380647.4±131145.9、278967.1±127630.7，B组低于A组（t=3.165，P=0.000），C、D、E组均高于B组（t值分别为5.272，7.132和2.950，P值均＜0.05），D组高于C组，但差异无统计学意义（t=1.953，P＞0.05），D组高于E组，差异有统计学意义（t=4.182，P＜0.05）。结论孕鼠补充牛磺酸可促进FGR胎鼠神经元和神经干细胞增殖，在妊娠第11天时补充效果最显著，可能发挥促进脑发育的最佳效应。

摘要： 目的：观察美满霉素对阿尔茨海默病（AD）大鼠认知功能和海马脑源性神经营养因子（BDNF）、凋亡相关因子Bcl-2和Bax表达的影响，探讨美满霉素对AD大鼠脑保护作用的机制。方法侧脑室注射Aβ25-35建立AD大鼠模型。30只健康雄性SD大鼠随机分成对照组、模型组和治疗组，每组10只。对照组和模型组腹腔内注射生理盐水1 mL/（kg·d），治疗组腹腔注射美满霉素50 mg/（kg·d），均持续14 d。Morris水迷宫检测行为学变化，蛋白免疫印迹（Western blotting）法和酶联免疫吸附试验（ELISA）法检测海马BDNF、Bcl-2和Bax蛋白的表达，原位末端标记（TUNEL）法检测海马神经元凋亡率。结果美满霉素可以明显提高AD大鼠学习记忆能力，上调AD大鼠海马BDNF、Bcl-2表达，下调Bax表达，减少海马神经元凋亡。结论美满美素可以通过促进神经元的生长、抑制神经元的凋亡发挥脑保护作用。%Objective To investigate the effect of minocycline on the cognition and expressions of brain-derived neurotrophic factor (BDNF), apoptosis related factor Bcl-2 and Bax in hippocampus of rats with Alzheimer’s disease (AD). Methods The rat model was established by microinjection of Aβ25-35 into lateral ventricle. Thirty healthy male SD rats were randomly divided into three groups:control group, model group and minocycline treatment group. Normal saline 1 mL/(kg·d) was intraperitoneally injected in control group and model group. The minocycline treatment group was intraperitoneally injected with minocycline 50 mg/(kg · d) for 14 days. Morris water maze was used to detect the behaviors of animals. The expressions of BDNF, Bcl-2 and Bax in hippocampus were measured by Western blotting and enzyme linked immunosorbent assay (ELISA). The apoptosis of neurons was detected by TdT-mediated dUTP nick-end labeling (TUNEL). Results Minocycline greatly improved the behaviors of AD rats, up-regulated the expressions of BDNF and Bcl-2, and down-regulated the expression of Bax in hippocampus, and reduced cell apoptosis. Conclusion Minocycline plays a protective role in neural function by promoting the growth of neurons and inhibiting the neuronal apoptosis.

摘要： 目的 评价依托咪酯后处理对大鼠缺氧缺糖-复糖复氧皮质神经元线粒体通透性转换孔(mPTP)的影响及其与Robo受体的关系.方法 出生24 h内的SD大鼠,体外培养其皮质神经元并接种于6孔板(2 ml/孔),采用随机数字表法分为4组(n=24):对照组(C组)、缺氧缺糖-复糖复氧组(OGD/R组)、依托咪酯后处理组(E组)和依托咪酯后处理+Robo受体阻断剂组(ER组).采用缺氧缺糖90 min,复氧复糖24 h的方法制备神经元缺氧缺糖-复糖复氧损伤模型.E组和ER组复氧复糖即刻于培养基中加入依托咪酯,终浓度6 μmol/L;ER组缺氧缺糖前6h时于培养基中加入Robo阻断剂RoboN,终浓度为1μg/ml.采用Hoechst/PI双染色法测定神经元凋亡情况,采用MTT法测定神经元存活情况,采用比色法测定LDH漏出情况;提取线粒体,测定线粒体通透性转换孔(mPTP)开放程度.结果 与C组比较,OGD/R组、E组和ER组神经元凋亡率、LDH漏出率和mPTP开放程度升高,神经元存活率降低(P＜0.05);与OGD/R组比较,E组神经元凋亡率、LDH漏出率和mPTP开放程度降低,神经元存活率升高,ER组神经元凋亡率和LDH漏出率降低,神经元存活率升高(P＜0.05);与E组比较,ER组神经元凋亡率、LDH漏出率和mPTP开放程度升高,神经元存活率降低(P＜0.05).结论 依托咪酯后处理通过激活Robo受体,抑制mPTP的开放,减轻大鼠皮质神经元缺氧缺糖-复糖复氧损伤.%Objective To evaluate the role of etomidate post-conditioning on mitochondrial permeability transition pore (mPTP) in the rat cortical neurons subjected to oxygen-glucose deprivation and restoration (OGD/R) and the relationship with Robo receptors.Methods The cortical neurons obtained from Sprague-Dawley rats (＜ 24 h after birth) were cultured in vitro and seeded in 6-well plates (2 ml/well).The neurons were divided into 4 groups (n=24 each) using a random number table:control group (group C),OGD/R group,etomidate post-conditioning group (group E),and etomidate post-conditioning + Robo receptor blocker group (group ER).The neurons were subjected to O2-glucose deprivation for 90 min followed by restoration of O2-glucose supply for 24 h.In E and ER groups,etomidate was added to the culture medium with the final concentration of 6 μmol/L immediately after onset of O2-glucose supply.In group ER,Robo blocker RoboN was added to the culture medium with the final concentration of 1 μg/ml at 6 h before O2-glucose deprivation.The neuronal apoptosis was detected using Hoechst/PI double staining,the viability of neurons was measured by MTT assay,and the amount of lactic dehydrogenase (LDH) released was measured using colorimetric method.The mitochondria were extracted,and mitochondrial permeability transition pore (mPTP) opening was detected.Results Compared with group C,the apoptosis rate,amount of LDH released,and mPTP opening were significantly increased,and the cell survival rate was decreased in OGD/R,E and ER groups (P＜0.05).Compared with group OGD/R,the apoptosis rate,amount of LDH released,and mPTP opening were significantly decreased,and the cell survival rate was increased in group E,and the apoptosis and amount of LDH released were significantly decreased,and the cell survival rate was increased in group ER (P＜0.05).Compared with group E,the apoptosis rate,amount of LDH released,and mPTP opening were significantly increased,and the cell survival rate was decreased in group ER (P＜0.05).Conclusion Etomidate post-conditioning mitigates OGD/R-induced damage to the cortical neurons through activating Robo receptors and inhibiting mPTP opening in rats.

摘要： 目的 探讨慢病毒介导的artemin(ARTN)基因修饰的骨髓间充质干细胞(MSCs)对帕金森病模型大鼠潜在性治疗作用及对脑内相关蛋白表达的影响.方法 分离培养大鼠MSCs,将携带ARTN的慢病毒载体转染MSCs;6-羟多巴胺(6-OHDA)制备帕金森病大鼠模型,按随机数字表法随机分为假手术(Sham)组、帕金森病组、移植MSCs(MSCs)组、移植空病毒转染的MSCs(LV-MSCs)组、移植慢病毒介导的ARTN基因修饰的MSCs (LV-ARTN-MSCs)组,对帕金森病大鼠左侧纹状体进行细胞移植;术后2、4、6、8周,腹腔注射阿扑吗啡,进行行为学检测;采用Western印迹及免疫荧光法检测各组大鼠黑质酪氨酸羟化酶(TH)表达变化,移植ARTN修饰的MSCs在大鼠脑组织内表达;高效液相色谱仪检测各组大鼠纹状体内多巴胺、二羟基苯乙酸、高香草酸含量.结果 LV-ARTN-MSCs组在阿扑吗啡诱发下30 min内旋转圈数(179.33±18.74)明显低于帕金森病组、MSCs组和LV-MSCs组(235.83±18.95、203.67±11.50和206.33±11.86;q=8.828,P＜0.01;q=3.802,P＜0.05;q=4.219,P＜0.05).LV-ARTN-MSCs组大鼠黑质TH阳性细胞百分比(64.05％±5.49％)显著高于帕金森病组(34.18％±3.35％)、MSCs组(52.59％±4.48％)和LV-MSCs组(50.57％±4.41％;q=13.280、5.135、6.028,均P＜0.01),黑质TH蛋白表达也明显提高,同时慢病毒介导ARTN修饰的MSCs可以在帕金森病大鼠脑内存活至少6周,主要集中在移植侧的纹状体区.细胞移植8周后,MSCs组(2.34±0.54)、LV-MSCs组(2.28±0.45)、LV-ARTN-MSCs组(6.54±0.51)与帕金森病组(0.87±0.07)大鼠相比,纹状体多巴胺表达明显提高(q =5.233,P＜0.05;q=5.020,P＜0.01;q=20.190,P＜0.01),以LV-ARTN-MSCs组最明显.结论 纹状体移植慢病毒介导ARTN基因修饰的MSCs对帕金森病模型大鼠有一定的治疗作用.%Objective To investigate the potential therapeutic efficacy of lentivirus-mediated artemin (ARTN) gene modified bone marrow mesenchymal stem cells (MSCs) transplantation on the rat model of Parkinson' s disease (PD) and the effects on expression of brain-related proteins.Methods MSCs were isolated and cultured in vitro,transfected by recombinant lentiviral vectors carrying ARTN gene.The PD rat model established by 6-hydroxydopamine (6-OHDA) was randomly divided into 5 groups:Sham group,PD group,MSCs group,MSCs transfected with empty lentiviral vectors transplanted (LV-MSCs)group and MSCs transfected with recombinant lentiviral vectors carrying ARTN gene transplanted (LVARTN-MSCs) group.The MSCs,LV-MSCs and LV-ARTN-MSCs groups were transplanted into the left striatum of each rat model of PD and ethology tests in every group were made with intraperitoneal injection of apomorphine (APO) 2,4,6,8 weeks after transplantation.The expression of tyrosine hydroxylase (TH) protein in substantia nigra (SN) was measured by Western blotting and immunohistochemistry,and immunofluorescence showed ARTN gene modified MSCs expression in rat brain tissue.The levels of dopamine (DA),dihydroxy-phenylacetic acid and homovanillic acid in striatum of each group were detected by high performance liquid chromatography.Results After injection of APO,rotation frequency decreased in LV-ARTN-MSCs group,i.e.(179.33 ± 10.74) circles/30 min vs (235.83 ± 18.95),(203.67 ±11.50) and (206.33 ± 11.86) circles/30 min in PD,MSCs and LV-MSCs groups (q =8.828,P ＜ 0.01;q =3.802,P ＜ 0.05;q =4.219,P ＜ 0.05).The percentage of TH-positive cells in SN after cell transplantation was increased significantly in LV-ARTN-MSCs group (64.05％ ± 5.49％) when compared with PD group (34.18％ ±3.35％),MSCs group (52.59％ ±4.48％) and LV-MSCs group (50.57％ ± 4.41％),respectively (q =13.280,5.135,6.028,all P ＜0.01).At the same time,TH protein in SN after cell transplantation was also increased obviously in LV-ARTN-MSCs group.ARTN gene modified MSCs can survive for at least 6 weeks in the rat brain of PD,mainly concentrated in the transplantation side of striatum.Eight weeks later,the levels of DA in striatum after cell transplantation were elevated significantly in MSCs group (2.34 ± 0.54),LV-MSCs group (2.28 ± 0.45) and LV-ARTN-MSCs group (2.28 ± 0.45)when compared with PD group (0.87 ± 0.07) (q =5.233,P ＜ 0.05;q =5.020,P ＜ 0.01;q =20.190,P ＜ 0.01),and LV-ARTN-MSCs group showed the most significant improvement.Conclusion ARTN gene modified bone marrow MSCs transplanted into the striatum of brain may have therapeutic effects on rat models of PD.

摘要： Objective To investigate the characteristics and mechanisms of sleep disorders in anti-Ma2 associated encephalitis.Methods Report a case of anti-Ma2 associated encephalitis with excessive sleepiness as the initial and prominent symptom who was diagnosed in our hospital.The characteristics of this case were studied from several aspects such as clinical, imaging, polysomnography and cerebrospinal fluid, etc.Results This case was characterized mainly by the symptom of lethargy, together with memory loss, mental and behavior disorder and walking instability.MRI examination indicated the damage of limbic system, hypothalamus and upper brainstem.The polysomnography showed narcolepsy-like symptoms and rapid eye movement sleep behavior disorder.In addition, the level of hypocretin from cerebrospinal fluid remained normal (186 pg/ml) , the anti-Ma2 antibodies in blood and cerebrospinal fluid showed positive, and the lymphoma was identified by colon histopathology.As a result, this case was diagnosed as lymphoma with anti-Ma2 associated encephalitis.Conclusions The sleep disorders in anti-Ma2 associated encephalitis manifest a variety of forms including excessive sleepiness, narcolepsy-like symptoms and rapid eye movement sleep behavior disorder, which are associated with the structural damage in hypothalamus, brainstem and limbic system.Further, the incidence of such narcolepsy symptoms might involve other neurotransmitter systems other than hypocretin.%目的 分析抗Ma-2抗体相关脑炎患者睡眠特点,探索睡眠障碍的机制.方法 回顾我院确诊的以过度嗜睡为首发症状的抗Ma-2抗体相关脑炎1例,分析其临床、影像、多导睡眠监测、脑脊液、病理检查等特点,结合文献进行讨论.结果 本例以嗜睡为突出症状,伴记忆力减退、精神行为异常、行走不稳等,磁共振成像提示边缘系统、下丘脑、上位脑干损害,多导睡眠监测表现为发作性睡病样特点及快速眼动期睡眠行为障碍,脑脊液下丘脑分泌素水平正常(186 pg/ml),血及脑脊液抗Ma-2抗体阳性,结肠组织病理检查为淋巴瘤,确诊为淋巴瘤合并抗Ma-2抗体相关脑炎.结论 抗Ma-2抗体相关脑炎的睡眠障碍可以表现为过度嗜睡、发作性睡病样症状及快速眼动期睡眠行为障碍等多种形式,与下丘脑、脑干、边缘系统等睡眠相关结构损害有关,下丘脑分泌素之外的递质系统可能参与发作性睡病症状的发生.

摘要： 由下述氟树脂形成了与蛋白质或包含蛋白质的组合物接触的表面的容器及用于制造蛋白质或包含蛋白质的组合物的器材具有显著的低蛋白质吸附性，其中，所述氟树脂为选自四氟乙烯‑六氟丙烯系共聚物及四氟乙烯‑全氟烷基乙烯基醚系共聚物中的至少1种氟树脂，并且，所述氟树脂的熔点为320°C以下，所述氟树脂中的相对于每1×10

摘要： 本发明提供能减小使用了特异性地结合的物质的测定中的由肝型脂肪酸结合蛋白质引起的测定值的变动幅度的肝型脂肪酸结合蛋白质制剂、评价该制剂的方法、制作肝型脂肪酸结合蛋白质的校准曲线的方法、以及定量该蛋白质的方法。一种用使用了用10mM的氧化剂在25°C下进行1小时的氧化处理的肝型脂肪酸结合蛋白质制剂的测定值与使用了未进行上述氧化处理的肝型脂肪酸结合蛋白质制剂的测定值的比表示的氧化变动系数设定为1.4以下的肝型脂肪酸结合蛋白质制剂、用于该制剂的肝型脂肪酸结合蛋白质、编码该蛋白质的DNA、由该DNA转化得到的细胞、该蛋白质的制造方法、评价该制剂的方法、抑制使用该制剂的测定中的测定值的变动幅度的方法、制作该蛋白质的校准曲线的方法、以及定量该蛋白质的方法。

摘要： 本发明提供一类能够携带生物活性蛋白分子穿透细胞膜的非肽类多胍有机小分子或其药学上可接受的盐，所述非肽类多胍有机小分子具有式I所示的结构：

摘要： 本发明涉及一种检测化合物的方法，该化合物能够调节神经元一氧化氮合酶(nNOS)或其中一种变体与神经元一氧化氮合酶抑制蛋白质(PIN)的复合作用，其中：温育含有所述化合物、PIN蛋白质和nNOS蛋白质或其中一种变体的混合物，检测与对照值相比PIN蛋白质与nNOS蛋白质或其中一种变体形成复合物量的显著变化，由此推断当存在如上所述的显著变化时，上述化合物与PIN蛋白质之间或上述化合物与nNOS蛋白质或其中一种变体之间存在键合作用，这导致调节PIN蛋白质与nNOS蛋白质和其中一种变体的复合作用。

摘要： 本发明涉及由序列编号1的氨基酸序列构成的组织纤维化预防或者治疗用重组蛋白质，作为基于乳脂球表皮生长因子8(Milk fat globule‑EGF factor 8)(MFG‑E8)蛋白质重组的蛋白质，以及包含该重组蛋白质的组织纤维化预防或者治疗用组合物。据此，相比于现有的基于乳脂球表皮生长因子8(MFG‑E8)蛋白质重组的蛋白质，本发明可显著改善组织纤维化预防或者治疗功能水平，通过预防以及治疗组织纤维化的恢复水平将非常接近正常组织。

摘要： 本发明提供一种糖化蛋白质测定试剂，其至少包含Trinder’s试剂、4‑氨基安替比林、蛋白酶、蛋白酶的稳定剂以及亚铁氰化物，其中，至少Trinder’s试剂包含在含有Trinder’s试剂的部分组合物中，至少4‑氨基安替比林包含在含有4‑氨基安替比林的部分组合物中，蛋白酶的稳定剂是使亚铁氰化物与该蛋白酶的稳定剂混合后的氧化还原电位大于0.058V的稳定剂，氧化还原电位是包含蛋白酶的稳定剂和亚铁氰化物、不包含糖化蛋白质的反应体系的氧化还原电位。

摘要： 本发明提供能够以优异的安全性将蛋白质导入到细胞中的蛋白质导入方法。通过使目标蛋白质承载在由粘土矿物构成的蛋白质导入用载体上并将其添加到细胞中，可以将所述目标蛋白质导入到所述细胞内中。所述粘土矿物优选为层状粘土矿物，可以使用蒙脱石、蛭石、伊利石等。

摘要： 本发明涉及与粉碎或切断洋葱等植物时发生催泪成分的生成有关的表现出将1－丙烯次磺酸转变为催泪成分的作用的蛋白质或多肽、编码该蛋白质或多肽的DNA(催泪成分合成酶基因)、使用用于属于葱属植物的催泪成分合成酶基因分离的引物以及使用该引物分离该基因的方法、含有上述DNA的重组载体、包含含有上述DNA的重组载体的转化体、对用含有上述DNA的重组载体转化的宿主细胞进行培养制造具有催泪成分合成酶活性的蛋白质或多肽的方法、具有催泪成分合成酶活性的蛋白质或多肽、以及具有抑制具有催泪成分合成酶活性的蛋白质或多肽的mRNA翻译的功能的核酸分子。

摘要： 本发明提供了一种应用iTRAQ技术研究弓形虫慢性感染小鼠脑组织差异表达蛋白质组的方法，本发明还提供了一种与大脑发育调节蛋白相互作用的蛋白质或多肽、及其应用，本发明通过CoIP和Pulldown实验发现与大脑发育调节蛋白相互作用6个弓形虫蛋白，并进一步针对TgRPS14、TgH4基因进行基因敲除实验和神经行为学验证。本发明提供了的弓形虫慢性感染小鼠脑组织差异表达蛋白质组的方法能快速有效地进行蛋白质组学研究，有利于快速定位到具有研究价值或商业应用价值的检测靶标、防治药物靶点。

摘要： 本发明涉及一种与蛋白质类药用活性成分结合使用的配体，所述配体由牛磺胆酸和甘氨胆酸组成。本发明还提供一种口服蛋白质类药物制剂，具体为以所述配体与蛋白质类药用活性成分形成的结合物为中心、外侧包裹纳米脂质体的微粒；所述纳米脂质体包裹层可以增加对核心结构的保护，且当活性成分为胰岛素时可为胰岛素质粒接触受体时产生自磷酸化提供磷离子。本发明提供的以胰岛素为活性成分的口服蛋白质类药物制剂不但能调节血糖、血脂，主要的是能防治II型糖尿病的并发症；其生物利用度高于注射胰岛素，不会引起低血糖，更不会至低血糖引起脑死亡；采用口服方式，给药方便，给药后血糖平稳，对机体的内环境的稳定更有益，具有良好且广泛的应用前景。