运动训练对糖尿病大鼠糖代谢和下丘脑神经肽Y mRNA的影响

目的:探讨运动训练对糖尿病大鼠糖代谢及其下丘脑神经肽Y(NPY)mRNA表达的影响。方法:30只SD大鼠随机分为对照组、糖尿病组(由链脲佐菌素诱导)和糖尿病 运动训练组(糖尿病大鼠进行无负重游泳运动,第1天游泳10min,以后每天递增10min,直到每天60min,每周5天,共10周)。10周后检测各组大鼠安静时血糖、血清胰岛素含量及下丘脑NPY mRNA表达。结果:与对照组相比,糖尿病组大鼠血糖显著升高,血清胰岛素水平显著降低;与糖尿病组相比,糖尿病 运动训练组大鼠血糖显著下降,血清胰岛素水平显著上升。糖尿病组大鼠下丘脑NPY mRNA表达显著高于对照组,糖尿病 运动训练组大鼠下丘脑NPY mRNA表达显著低于糖尿病组。结论:适宜的运动训练可能通过下调下丘脑NPY mRNA表达来改善糖尿病大鼠糖代谢异常。     

麻杏二三汤对哮喘小鼠气道黏蛋白5ac mRNA影响的实验研究

目的观察麻杏二三汤对哮喘小鼠肺组织气道黏蛋白(MUC)5ac m RNA的影响,探讨其抑制哮喘小鼠气道黏液高分泌的作用机制。方法雌性BALB/c小鼠60只,随机分成空白对照组、哮喘模型组、强的松组及麻杏二三汤高、中、低剂量组,每组10只。除空白对照组,另五组以腹腔注射0.1%卵白蛋白(OVA)与10%氢氧化铝混悬溶液致敏,以1%OVA雾化液雾化攻击制作哮喘小鼠模型。分别给予生理盐水、生理盐水、强的松(10.0mg/kg)、麻杏二三汤高剂量(30g/kg)、中剂量(15g/kg)、低剂量(7.5g/kg)灌胃,R-T PCR测定各组MUC5ac m RNA表达。结果麻杏二三汤高、中、低剂量组MUC5ac m RNA荧光定量△Ct值分别为(12.80±0.84)、(12.15±0.49)、(11.41±1.49),均高于哮喘模型组的(9.82±0.84)(P0.01)。结论麻杏二三汤能够有效抑制哮喘小鼠气道黏液高分泌,其机制可能与其下调支气管肺组织中MUC5ac m RNA表达有关。     

肝癌病人外周血及骨髓AFP mRNA的检测及意义

目的 通过检测AFP mRNA在肝细胞性肝癌（hepatocellular carcinoma,HCC)病人外周血和骨髓中的表达，探讨AFP mRNA作为HCC微转移指标的可能性。方法 建立敏感的巢式逆转录聚合酶链反应体系（Nested-RT-PCR)，检测71例HCC、36例肝硬化、9例肝血管瘤、11例肝转移瘤、30例正常对照外周血AFP mRNA和19例HCC、15例血液病骨髓AFP mRNA。结果 71例HCC外周血中38例AFP mRNA表达阳性（53．5％），肝硬化、肝血管瘤、肝转移瘤和正常对照均为阴性。HCC外周血AFP mRNA检出率与血清AFP值无明显相关（P＞0．05），而与肿瘤大小、门静脉癌栓、肝内转移、肝外转移等临床参数有明显相关（P＜0．05）。19例HCC骨髓中9例AFP mRNA表达阳性（47．4％），15例血液病为阴性。骨髓AFP mRNA检出率与血清AFP值、肝内转移无明显相关（P＞0．05），与肿瘤大小、门静脉癌栓、肝外转移明显相关（P＜0．05）。外周血和骨髓AFP mRNA的表达呈一致性（P＜0．05）。结论 血循环AFP mRNA可作为CC微转移的早期检测指标。HCC病人外周血和骨髓AFP mRNA的表达与复发转移显著相关，且HCC病人外周血和骨髓AFP mRNA的表达呈一致性，联合检测有助于提高结论的可靠性。     

用SYBR Green I实时逆转录-聚合酶链反应定量检测人、小鼠精子中CatSper1 mRNA

目的:建立并优化SYBR GreenI实时RT-PCR体系,定量检测人、小鼠成熟精子中的CatSper1 mRNA。方法:用TRIzol分别提取人、小鼠成熟精子中的总RNA,逆转录后用SYBR GreenI实时PCR定量检测CatSper1 mRNA。SYBR GreenI实时PCR采用普通PCR试剂,加入SYBR GreenI染料,优化退火温度、Mg2+浓度及上、下游引物比例,并在PCR循环时采用四步法以消除引物二聚体的影响。优化完成后用不同浓度的精子cDNA为模板做标准曲线,以检测SYBR GreenI实时PCR的扩增效率。结果:定量检测CatSper1 mRNA的SYBR GreenI实时PCR体系适宜退火温度、Mg2+浓度及上、下游引物比例分别为63℃、3.0mmol/L和1∶1,四步法中采集荧光的温度为88℃。优化后用人和小鼠精子cDNA为模板做标准曲线分别为Y=-3.402log(X)+25.99和Y=-3.409log(X)+24.09,扩增效率分别为96.8%和96.5%,可定量检测人、小鼠成熟精子中的CatSper1 mRNA。结论:用普通的逆转录及PCR系统和试剂,建立了一种方便、廉价、可靠的SYBR GreenI实时荧光定量RT-PCR系统,可用于人、小鼠精子中CatSper1 mRNA定量检测。     

Wnt-1诱导分泌蛋白1在直肠癌中的表达及其临床意义

目的定量检测Wnt-1诱导分泌蛋白1（Wnt-1 induced secreted protein-1，WISP-1）基因在人直肠癌及远端正常直肠组织中的表达，为进一步研究该基因功能提供线索。方法提取人直肠癌及癌旁正常直肠组织总RNA，采用实时RT—PCR分析WISP-1的表达，再对PCR结果进行分析。结果WISP-1 mRNA在直肠癌组织中的表达水平显著高于癌旁正常直肠组织，差异有统计学意义（P〈0．05）。WISP-1的表达与肿瘤的分化程度、Dukes分期及淋巴结转移等因素有相关性。结论WISP-1基因的异常表达可能与直肠癌的发生、发展有关，有望成为直肠癌诊断预后的标志物和治疗的新靶点。     

Age-related effects of estrogen on the expression of estrogen receptor (ER) α and β mRNA in the ovariectomized (OVX) monkey hypothalamus

In the present study, we reported distribution of ERα and ERβ mRNAs in the hypothalamus of young and old ovariectomized (OVX) rhesus macaques. The ERα were detected in all six major vestiblular nuclei which included arcuate nucleus (ARC) , paraventricularis nucleus (PVN) , periventricular nucleus (PeriV) , supraoptic nucleus (SON) , medial prioptic nucleus (MPN) and lateral hypothalamus area (LHA). However, the ERβ mRNA can also detected in those nuclei excerpt SON, but the signals of ERβ mRNA were weaker than those of ERα mRNA. We observed that the degree of expression of ERs mRNA were different in most nucleus of old and young monkeys. The ERα mRNAs were highly expressed in ARC and SON in young monkeys compared with old monkeys. Moderate amount of ERα mRNAs hybridization signals and weak signals were observed in LHA, and MPN both in young and old monkeys. In contrast, only lower level of ERα hybridization signal were observed in PVN and PeriV in young monkeys, and the signals of ERα were very low in those nucleus of old monkeys. In general, the expression of ERβ mRNA were weaker than that of ERα mRNA in above nucleus excerpt LHA. The relatively higher density of ERβ hybridization signals have been observed in the LHA in young monkey compared with old monkeys. Low amount of ERβ mRNA hybridization signals were observed in the ARC, PVN and MPN, and no age differences were seen in PVN and MPN of those monkeys. In PeriV, we observed some signals in young monkey and a few signals in old monkeys. It was different from the rodent in which we did not found ERβ hybridization signal in SON. This study showed that both of the two estrogen receptors not only had the same pattern of expression but also had many different patterns of expression. The different expression of ERα and ERβ mRNAs in the young and old monkey brain may imply diverse functions in different regions of the monkey brain.     

神经生长因子和脑源性神经营养因子在脑胶质瘤内的表达

目的研究NGF(NeverGrowth Factor,神经生长因子)、BDNF(BrainDerivedNeurotrophicFactor,脑源性神经营养因子)在脑胶质瘤内的表达,探讨它们与肿瘤病理分级、实体肿瘤部位的关系,以及它们在肿瘤发生过程中的作用。方法通过免疫组织化学SP染色方法检测70例胶质瘤和5例正常脑组织中NGF和BDNF的表达。结果NGF和BDNF阳性细胞表达率明显高于正常脑组织内的表达,有统计学意义(P<0.05);在不同病理级别胶质瘤之间阳性细胞表达率也有差异,分别是:I级16.56%,11.24%;II级32.45%,18.23%;III级40.91%,21.44%;IV级24.71%,15.39%,正常脑组织内的表达率为7.06%,5.98%,且有统计学意义(P<0.05)。NGF和BDNF阳性细胞表达率在不同部位胶质瘤内各不相同,分别为:颞叶42.57%,24.19%;顶叶35.62%,20.09%;小脑28.67%,16.17%;额叶21.45%,12.42%,差异有显著性(P<0.05)。结论NGF和BDNF在脑胶质瘤内高表达;NGF和BDNF阳性细胞表达与胶质瘤的病理级别、肿瘤部位有关;NGF和BDNF可能参与了胶质瘤的发生。     

谷氨酰胺对2型糖尿病大鼠血糖和胰岛素抵抗的影响

目的 观察L-谷氨酰胺对2型糖尿病大鼠血糖变化和胰岛素抵抗的影响并探讨可能机制.方法 SD雄性大鼠47只,按随机数字表法分为4组:空白对照组(C组)10只、谷氨酰胺(glutamine,Gln)组(Gln组)10只、2型糖尿病组(DM组)12只、2型糖尿病+谷氨酰胺组(DM+Gln组)15只.实验动物分组饲养,2型糖尿...     

AFP mRNA在监测肝癌转移复发上的研究现况

手术切除是目前治疗肝癌的最佳途径,但术后肿瘤的转移复发严重制约了手术的疗效.肝癌的转移和复发不但严重威胁着人们的健康,而且给患者家庭以及社会都带来了沉重的负担.当前公认肝癌的诊断标志物为AFP,但其特异性不高,且AFP的生物学特性使其不能作为监测肝癌转移复发的标志物.随着基因技术的发展,人们发现AFP的前体AFP mR...