左旋多巴对帕金森病大鼠模型的脑保护作用

目的 探讨左旋多巴对不同程度损伤巴金森病（PD）模型大鼠脑的影响。方法 采用左旋多巴与生理盐水处理不同程度损伤PD鼠及PD模型假手术鼠，观察各实验组鼠行为、纹状体区谷胱甘肽的含量、黑质区TH神经元和纹状体区TH神经纤维数的变化。结果 左旋多巴治疗诱发的旋转行为只见于重度损伤PD鼠。左旋多巴对中、重度损伤PD鼠纹状体区的谷胱甘肽水平有上调作用，分别达16．3％和19．6％（P＜0．05），而轻度损伤PD鼠谷胱甘肽变化不明显。左旋多巴增加中、轻度损伤PD鼠纹状体TH阳性神经纤维数分别达93．5％和40．8％（P＜0．05）。结论 左旋多巴对中、重度损伤PD大鼠纹状体区谷胱甘肽水平有上调作用；左旋多巴升高纹状体区谷胱甘肽水平，与PD大鼠的损伤程度不相关；左旋多巴可促进轻、中度损伤PD鼠纹状体区TH阳性神经纤维的恢复。     

针刺对帕金森模型大鼠纹状体多巴胺及代谢产物含量的影响

目的探讨针刺治疗帕金森病（PD）模型大鼠的疗效及作用机制.方法应用偏侧纹状体立体定向注射6-羟基多巴胺的方法制备PD模型大鼠,针刺百会、风府和双侧阳陵泉治疗,观察针刺前后PD模型大鼠阿朴吗啡诱导的行为学及纹状体多巴胺（DA）及代谢产物含量的变化.结果与模型组比较,针刺后PD大鼠行为学明显改善（P〈0.01）,同时针刺组大鼠纹状体DA及代谢产物的含量均显著增加（P〈0.01）.结论针刺可明显改善PD模型大鼠的行为学,并且可以提高纹状体DA及其代谢产物的含量.     

三七总皂甙对神经干细胞诱导分化多巴胺能神经元移植帕金森病大鼠的影响研究

目的 探讨三七总皂甙对神经干细胞体外诱导分化多巴胺能神经元移植帕金森病(PD)大鼠后移植细胞的存活及移植疗效的影响.方法 大鼠胚胎中脑神经干细胞经传代扩增后,在分化液中诱导向多巴胺能神经元分化,应用6-羟基多巴胺制备的PD大鼠模型随机分为多巴胺能神经元组、多巴胺能神经元 三七总皂甙组、三七总皂甙组及手术对照组,每组8只,进行移植手术,移植后检测PD大鼠不对称旋转行为的变化及纹状体移植区酪氨酸羟化酶染色阳性细胞存活的情况.结果 与手术对照组比较,移植后20 d多巴胺能神经元组大鼠不对称旋转圈数开始明显下降(P＜0.01).移植后10～60 d,多巴胺能神经元 三七总皂甙组大鼠的不对称旋转圈数明显低于多巴胺能神经元组(P＜0.01).免疫组织化学染色显示多巴胺能神经元 三七总皂甙组大鼠纹状体移植区酪氨酸羟化酶染色阳性细胞数明显多于多巴胺能神经元组(P＜0.01).结论 三七总皂甙具有提高神经干细胞诱导分化的多巴胺能神经元移植PD大鼠后移植细胞的存活率及移植疗效的作用.     

99mTc-TRODAT-1放射自显影对帕金森病大鼠模型多巴胺转运蛋白的观察

目的　探讨 99m Tc- TRODAT- 1多巴胺转运蛋白 (DAT)显像临床应用价值。　方法　应用 6 -羟基多巴胺 (6 - OHDA)建立完全损毁及部分损毁一侧帕金森病 (PD)大鼠模型 ,以 99m Tc-TRODAT- 1作为配体 ,采用放射自显影观察一侧 PD大鼠模型 DAT分布及其密度 ,高效液相 -电化学方法检测模型大鼠纹状体多巴胺 (DA)及其代谢产物含量 ,免疫组化酪氨酸羟化酶 (TH)染色观察模型大鼠黑质及纹状体 TH阳性细胞及纤维。　结果　 6 - OHDA损毁侧纹状体放射性浓集明显低于未损毁侧 ,完全损毁模型的纹状体放射性浓集最低。纹状体 DA含量部分损毁及完全损毁较未损毁侧分别降低 39%和 98%。TH染色可见损毁侧黑质及纹状体 TH阳性细胞及纤维明显少于对侧。　结论　 PD大鼠模型损毁侧纹状体 DAT密度降低 ,且与损毁程度有关。99m Tc- TRODAT- 1DAT显像研究可能有助于 PD的早期诊断。     

基因治疗帕金森病大鼠脑内纹状体多巴胺含量的检测

目的观察脑源性神经营养因子（ＢＤＮＦ）基因工程成肌细胞脑内纹状体移植对帕金森病（ＰＤ）大鼠脑内纹状体多巴胺含量的影响。方法建立逆转录病毒介导的ＢＤＮＦ表达质粒,并转染成肌细胞进行ＰＤ大鼠脑内纹状体移植。结果细胞移植后第２和第８周,移植组毁损侧纹状体多巴胺含量〔分别为（９５７５３±８８９５）和（１０４０２９±１０４７８）ｐｇ／ｍｇ〕较对照组〔分别为（３３５９８±１０２４８）和（３２７８８±７０２３）ｐｇ／ｍｇ〕明显增加（均为Ｐ＜００１）,并可维持２个月之久。结论脑源性神经营养因子基因工程成肌细胞脑内纹状体移植,可使脑内纹状体多巴胺含量明显增加,可能为帕金森病的治疗提供了一种新的治疗方法。     

^99mTc—TRODAT—1放射自显影对帕金森病大鼠模型多巴胺？…

目的 探讨＾９９ｍＴｃ－ＴＲＯＤＡＴ－１多巴胺转运蛋白（ＤＡＴ）显像临床应用价值。方法 应用６－羟基多巴胺（６－ＯＨＤＡ）建立完全损毁及部分损毁一侧帕金森病（ＰＤ）大鼠模型，以＾９９ｍＴｃ－ＴＲＯＤＡＴ－１作为配体，采用放射自显影观察一侧ＰＤ大鼠模型ＤＡＴ分布及其密度，高效流体相电化学方法鼠黑质及纹状体ＴＨ阳性细胞及纤维。结果 ６－ＯＨＤＡ损毁侧纹状体放射性浓集明显低于本损毁侧，完全损毁模型的纹状     

腺病毒介导的胶质细胞源性神经营养因子基因脑内转移对帕金森病大鼠模型的保护作用

目的 探讨腺病毒介导的胶质细胞源性神经营养因子（GDNF）基因直接转移对帕金森病（PD）的保护作用。方法 实验SD大鼠分重组GDNF腺病毒（Ad－GDNF）实验组、LacZ腺病毒（AdLacZ）对照组和磷酸缓冲液（PBS）对照组。将重组腺病毒及PBS定向注射至一侧黑质附近,1周后于同侧纹状体注射6－羟基多巴胺（6－OHDA）诱发多巴胺（DA）能神经元进行性变性。通过旋转行为观察和中脑酪氨酸羟化酶（TH）免疫组化染色及纹状体单胺类递质高压液相色谱－电化学仪（HPLC－ECD）检测评估其治疗效应;通过RT－PCR、ELISA检测Ad-GDNF在脑内的表达情况。结果 Ad-GDNF组阿扑吗啡诱发的旋转次数明显低于2个对照组;Ad-GDNF组约70％的黑质DA能神经元得以保存,而Ad-LacZ及PBS对照组令有30％左右;Ad－GDNF组纹状体DA含量也显著高于Ad-LacZ及PBS对照组;Ad－GDNF在脑内可有效表达,注射后5周黑质附近GDNF含量达1ng/10mg脑组织湿重,是对照组的16－20倍。结论 腺病毒介导的GDNF基因脑内直接转移可阻止6－OHDA诱发的大鼠DA能神经元进行性变性,提示这一手段在PD保护性治疗方面具有一定的应用价值。     

羊膜上皮细胞移植治疗帕金森病大鼠的实验研究

目的 探讨羊膜上皮细胞纹状体内移植对帕金森病（PD）大鼠的治疗作用。方法 采用RT-PCR方法检测入羊膜上皮细胞株FL表达脑源性神经营养因子（BDNF）、神经营养因子-3（NT-3）mRNA;Hoechest33342标记后微移植方法将其移入PD大鼠模型纹状体内，免疫荧光技术检测移植细胞表达BDNF、NT-3状况。结果 羊膜上皮细胞能够表达BDNF、NT-3，植入纹状体内能够存活，并在第2周内明显改善PD大鼠的行为学症状。结论 羊膜上皮细胞可作为表达神经营养因子治疗PD的移植细胞。     

酪氨酸羟化酶cDNA移植治疗帕金森病的实验研究

目的 评价质粒ＤＮＡ脂质体复合物直接转染纹状体细胞表达酪氨酸羟化酶治疗帕金森病的疗效。 方法 应用６－羟多巴胺制备偏侧帕金森病大鼠模型，首次将表达大鼠酪氨酸羟化酶的质粒ＤＮＡ脂质体复合物植入大鼠损毁侧纹状体。移植后，观察大鼠模型旋转行为的改善程度，并用免疫组化及逆转录聚合酶链反应（ＲＴ－ＰＣＲ）检测酪氨酸羟化酶基因的表达。 结果 移植后２周大鼠模型的旋转行为明显改善（治疗后３、６、９、１２及１５天     

纹状体神经元GluR1Ser831磷酸化对帕金森病异动症发病机制的影响

目的 从纹状体神经元谷氨酸受体1的831位丝氨酸（GluRlSer831）磷酸化角度，探讨长期左旋多巴治疗帕金森病（PD）的异动症发病机制。方法 通过6羟基多巴胺立体定向注射至大鼠前脑内侧束建立PD动物模型，然后左旋多巴甲酯腹腔注射治疗（25mg·kg^-1·d^-1，每天2次）22d，评估左旋多巴处理剂峰旋转行为情况；运用免疫荧光与免疫印迹法（Western blot）观察和检测纹状体区谷氨酸受体GluR1亚细胞分布及GluR1Ser831磷酸化的表达情况。结果 PD大鼠应用左旋多巴长期处理后出现明显的剂峰旋转行为增强，与PD患者异动症具有相似特征。PD大鼠损伤侧纹状体细胞膜上GluR1和GluR1Ser831磷酸化的数量分别减少至（73．0±4．8）％和（51．4±4．3）％；长期左旋多巴处理又使损伤侧纹状体细胞膜上GluR1和GluR1Ser831磷酸化的数量分别增加至（104．2±5．5）％和（93．7±3．1）％；而损伤侧纹状体总蛋白GluR1的数量未发生明显变化。GluR1和GluR1Ser831磷酸化的这些改变独特发生在纹状体小清蛋白阳性中间神经元上。结论 纹状体小清蛋白阳性中间神经元上GluR1及GluR1Ser831磷酸化的改变，可能与PD异动症的发生有关。     

树突状细胞诱导的抗肿瘤免疫诱导移植瘤细胞凋亡并抑制其增殖

目的研究树突状细胞(DC)体外诱导的细胞免疫能否抑制裸鼠移植瘤生长及其机制.方法联合应用粒/巨噬细胞集落刺激因子(GM-CSF)及白介素-4(IL-4)直接从肝癌患者外周血中培养出DC,以源于人肝癌细胞系HepG2肿瘤细胞的肿瘤抗原粗提物刺激DC,DC激活同源的T淋巴细胞产生细胞毒性T淋巴细胞(CTL),建立裸鼠人肝癌细胞系HepG2移植瘤模型.以CTL治疗裸鼠HepG2移植瘤并观察治疗效果,检测移植瘤标本肿瘤细胞凋亡情况.结果DC诱导的CTL通过诱导肿瘤细胞凋亡并抑制其增殖而抑制移植瘤生长.结论经肿瘤抗原激发的DC有可能在肿瘤的治疗中发挥重要作用.     

Attempted enzyme replacement using human amnion membane implantations in mucopolysaccharidoses

Summary Amnion membrane implantation has been proposed as an approach to enzyme replacement in mucopolysaccharidoses. Human amnion membranes have been subcutaneously implanted in the abdominal wall in 19 patients with mucopolysaccharidoses (MPS I, II and III). A protocol was developed for the objective evaluation of experimental treatments of these patients. Systematic evaluation of the clinical status before and 6 months after amnion membrane implantation reveals no change in function except improvement in joint mobility. The sum of all joint movements showed improvement from baseline values to 6 months after implantation by ANOVA followed bypost-hoc analysis (p<0.056). The only specific joint movements to significantly improve after 6 months were shoulder extension (p<0.01) and hip internal rotation (p<0.05). Serial measurements of the deficient lysosomal enzyme activity in serum and white blood cells did not increase in any patient after amnion membrane implantation. Urinary glycosaminoglycan excretion decreased transiently in 2 of 10 patients after implantation, but a second amnion membrane implantation did not result in any change. Biopsy of the implantation site in 10 patients 6 months after amnion membrane implantation revealed a foreign-body reaction with giant cell formation and fibrosis and no recognizable amnion membrane tissue. We conclude that human amnion membrane implantation is not an effective therapy in mucopolysaccharidoses.     

Effect of the suckling stimulus on daily LH surges induced by chronic oestrogen treatment in ovariectomized lactating rats

Effects of the suckling stimulus on the daily LH surge induced by chronic oestrogen treatment were examined in ovariectomized lactating rats. Wistar-Imamichi strain rats were kept under 14 h light:10 h darkness (lights on at 05.00 h). Litter size was adjusted to eight on day 1 (day 0 = day of parturition) and ovariectomy performed on day 2. Lactating rats deprived of their litters on day 0 served as nonlactating controls. Silicone elastomer tubing filled with oestradiol was implanted on day 6 or 15. Blood samples were collected through an indwelling cannula at 10.00 and 17.00 h on each day after implantation to detect daily LH surges. Daily LH surges occurred in the late afternoon in both lactating and non-lactating rats implanted with oestradiol on day 6 or 15. The amplitude of daily LH surges in lactating rats implanted on day 6 declined much more rapidly than in non-lactating rats implanted on day 6, but no significant difference was found in the profile of the LH surge between lactating and non-lactating rats implanted on day 15. Pituitary LH contents just before the daily LH surge (12.00-12.30 h) 4 days after implantation in lactating rats implanted with oestradiol on day 6 were significantly less than those in nonlactating rats implanted with oestradiol on day 6 or 15 and in lactating rats implanted on day 15.(ABSTRACT TRUNCATED AT 250 WORDS)     

Steroid sulfatase activity in amnion tissue and amnion cells maintained in monolayer culture

Phenolic steroid sulfatase activity in amnion tissue, amnion homogenate, subcellular fractions, and amnion epithelial cells in culture was demonstrated with radiolabeled estrone sulfate as the substrate. Sulfatase activity could not be detected in either amnion tissue or cells when evaluated with dehydroisoandrosterone sulfate as the substrate. Phenolic steroid sulfatase activity in amnion tissue was linear with incubation time up to 3 h and with amnion tissue weight up to 800 mg/ml. The rate of estrone sulfate hydrolysis in amnion tissue increased in a linear manner with temperature from 3 to 60 C. The apparent Km of amnion tissue sulfatase for estrone sulfate was 9 microM. The highest specific activity of the enzyme was found in both the mitochondrial-lysosomal and microsomal fractions. In studies with amnion epithelial cells in monolayer culture, phenolic steroid sulfatase activity was linear with incubation time up to 4 h and with cell number up to 2 X 10(5)/ml. The apparent Km of amnion cell sulfatase for estrone sulfate was 5.5 microM. The product of hydrolysis, i.e. estrone, was metabolized in situ to 17 beta-estradiol in both amnion tissue and cells. The hydrolysis of estrone sulfate (and possibly other phenolic steroid sulfates present in amniotic fluid) by amnion cells may be important in providing biologically potent estrogens for in situ action.     

缺氧预处理提高骨髓基质细胞促进血管新生能力

目的 研究缺氧对骨髓基质细胞(bone marrow stromal cells，BMSCs)表达血管内皮细胞生长因子(vascular endothelial growth factor，VEGF)及促进血管新生能力的影响。方法 将BMSCs置于缺氧环境下培养24h后，研究其VEGF表达的变化。将急性心肌梗死大鼠随机分为3组。未处理组在梗死后4w移植未经缺氧处理的BMSCs；缺氧组将经缺氧预处理的BMSCs注射到心肌梗死区；对照组仅注射无血清的培养液。在梗死后6w取各组标本观察梗死区及周围VEGF的表达和血管新生状况。结果 体外培养的BMSCs经缺氧预处理后VEGF的表达明显增加。缺氧组和未处理组梗死区及周围VEGF的表达和毛细血管密度较对照组明显增加。缺氧组VEGF的表达和毛细血管密度又较未处理组明显增加。结论 体外缺氧可诱导BMSCs高表达VEGF。BMSCs移植后促进梗死区及周围VEGF的表达，对血管新生有积极作用。移植前缺氧预处理使BMSCs促进缺血心肌组织表达更多的VEGF从而提高其促进血管新生作用。     

GH3 pituitary adenoma cells can reverse thymic aging in rats.

Thymic size and T-cell function decrease with age, and it has not yet been possible to totally reverse this thymic atrophy and completely restore T-cell-dependent immune functions. In this study, GH3 pituitary adenoma cells, which secrete growth hormone and prolactin, were implanted subcutaneously into 16- and 22-month-old female Wistar-Furth rats and the rats were sacrificed approximately 2 months later. Only thymic remnants were detected in aged, non-implanted rats, but thymus glands were found in both the 18- and the 24-month-old rats that had been implanted with GH3 cells. Thymus glands from the GH3-implanted 18-month-old rats contained distinct cortical thymocytes and medullary epithelial cells. Depending on the concentration of phytohemagglutinin or concanavalin A, T-cell proliferative responses of splenocytes from these implanted rats were 2- to 5-fold greater than those of 18-month-old controls. At the optimal concentration of mitogen, proliferative responses to either lectin could be restored to those levels observed in splenocytes from 3-month-old Wistar-Furth females. Thymus glands from 24-month-old GH3-implanted rats contained more cortical thymocytes and fewer fat vacuoles than controls, but they were not totally reconstituted. No significant lectin-induced T-cell proliferative responses or IL-2 secretion were detected in 24-month-old control rats, but splenocytes from GH3-implanted rats showed augmented T-cell proliferative responses and increased synthesis of IL-2. Fluorescence-activated cell-sorter analysis of thymocytes revealed that 24-month-old rats implanted with GH3 cells had a higher proportion of lymphocytes with the Thy-1.1 and helper-T-cell phenotypes. These data show that it is possible to regenerate normal thymic tissue in situ and reverse the natural loss in cell-mediated immunity that occurs with aging.     

Enhancement of glucocorticoid-induced 11beta-hydroxysteroid dehydrogenase type 1 expression by proinflammatory cytokines in cultured human amnion fibroblasts

Glucocorticoids and proinflammatory cytokines may be involved in parturition by stimulation of prostaglandin production in the fetal membranes. The actions of glucocorticoids on the fetal membranes are amplified by 11beta-hydroxysteroid dehydrogenase type 1 (11beta-HSD1), which converts biologically inactive cortisone into active cortisol. Whether glucocorticoids and proinflammatory cytokines regulate the expression of 11beta-HSD1 in the major prostaglandin-producing tissue, amnion, thus further increasing prostaglandin production, is not known. In this study, we found that term amnion fibroblasts had higher 11beta-HSD1 mRNA and activity per cell than amnion epithelial cells. Both isoforms of glucocorticoid receptor (alpha and beta) were expressed in amnion fibroblasts and epithelial cells. Quantitative real-time PCR showed that dexamethasone (0.01-1 microm) dose-dependently induced 11beta-HSD1 mRNA expression only in amnion fibroblasts but not in amnion epithelial cells. The induction of 11beta-HSD1 mRNA expression by dexamethasone was blocked by glucocorticoid receptor antagonist RU486. Although only a modest increase or no change in 11beta-HSD1 mRNA expression and activity was observed with IL-1beta (10 ng/ml) or TNFalpha (10 ng/ml) treatment, respectively, in amnion fibroblasts, combination of dexamethasone with either IL-1beta or TNFalpha significantly enhanced the induction of 11beta-HSD1 mRNA expression and activity, as compared with dexamethasone treatment alone. With prior induction of 11beta-HSD1 expression by dexamethasone, cortisone caused more prostaglandin E2 production in the amnion fibroblast. This study suggests that glucocorticoids can positively induce 11beta-HSD1 expression in amnion fibroblasts, an effect further strengthened by proinflammatory cytokines.     

磁共振成像示踪观察脑缺血大鼠脑室内移植神经干细胞的实验研究

目的探讨MRI示踪观察脑缺血大鼠脑室内移植的标记神经干细胞的分布和迁移的可行性。方法选取16只雄性SD大鼠制做脑梗死模型,随机分为标记组和未标记组,每组8只,分别于脑梗死对侧侧脑室内立体定向移植超顺磁性氧化铁纳米颗粒标记和未经标记的神经干细胞。移植细胞后,不同时间点分别进行MRI连续成像观察,之后进行大鼠脑组织冷冻切片的普鲁士蓝染色。结果标记组移植后即刻的MRI即可观察到大鼠侧脑室内的移植标记细胞,移植后7天脑梗死皮质下即可见到低信号影,各对应时间点组织切片的普鲁士蓝染色与MRI结果相符。结论MRI能够在活体内连续无创的示踪观察大鼠侧脑室移植的标记神经干细胞的迁移及分布情况。     

内皮祖细胞移植治疗大鼠急性心肌梗死的实验研究

目的探讨大鼠内皮祖细胞(EPCs)移植于梗死心肌的增殖分化情况及对心功能的影响。方法分离培养大鼠EPCs,免疫荧光法检测其CD34+、CD133+和Flk-1+的表达。将SD大鼠冠状动脉左前降支结扎制造急性心肌梗死模型后,在梗死心肌处植入DAPI标记的EPCs(实验组)或M 199培养液(对照组)。移植后1周及4周,心脏超声检查心功能,并对梗死区心肌组织进行移植细胞形态学检查及毛细血管密度测定,逆转录聚合酶链反应及酶联免疫吸附测定梗死周边区血管内皮生长因子(VEGF)的表达。结果培养获得EPCs,其表型为CD34+、CD133+和Flk-1+。植入梗死区的EPCs可以分化为血管内皮细胞,实验组梗死心肌处血管密度较对照组明显增高(P<0.01),并且VEGF基因及蛋白表达在移植后1周均较对照组明显增高(P<0.01)。移植后4周,实验组大鼠左心室射血分数及左心室短轴缩短率较对照组明显提高(P<0.01);而移植后1周,两组大鼠超声检测心功能各指标变化不明显。结论同种异体EPCs移植到梗死心肌大鼠缺血心肌能分化为毛细血管内皮细胞,促进梗死后心肌血管新生,改善心功能。     

白芨多糖对心肌梗死大鼠PI3K/AKT/GRP78信号通路及心肌细胞内质网应激凋亡的影响

目的 探究白芨多糖对心肌梗死(MI)大鼠磷脂酰肌醇-3-激酶(PI3K)/蛋白激酶B(AKT)及葡萄糖调节蛋白(GRP)78信号通路蛋白表达及对心肌细胞内质网(ERS)凋亡的影响.方法 SD大鼠随机分为假手术组、模型组、白芨多糖低(100 mg/kg)、中(200 mg/kg)、高(400 mg/kg)剂量组、阳性对照...