骨髓基质细胞促进神经干细胞增殖分化

目的：探讨骨髓基质细胞（BMSCs）对神经干细胞（NSCs）增殖分化的影响。 方法:在体外比较NSCs在单独培养和在BMSCs条件培养液中培养下的分化和增殖情况。 结果:应用BMSCs条件培养液培养NSCs，分化的神经元比例较显著高于NSCs单独培养（41.1%±3.2% vs 23.3%±16.5%，P<0.05），而分化的星形胶质细胞所占比例显著降低（33.8%±4.9% vs 65.0%±10.4%，P<0.01），同时增殖细胞所占比例也显著增高（74.7％±4.7％ vs 51.4％±12.3％，P<0.01）。 结论:BMSCs对NSCs有促进其增殖和向神经元分化的作用。NSCs与BMSCs联合移植可能会增强NSCs移植的抗脑损伤作用。     

B群人格障碍的人格五因素特征

目的:了解B群人格障碍的人格五因素特征。方法:先用人格障碍诊断问卷第四版(PDQ 4)在被试中筛查,再用个性障碍晤谈手册(PDI-IV)做半定式查询确定B群人格障碍患者,用NEO个性问卷(NEO-PI-R)做问卷测查。结果:66例B群人格障碍患者表现为高神经质(61.97±10.57)、低顺同性(38.39±11.46)和低严谨性(38.40±11.19),外向性(45.88±11.52)和开放性(45.00±9.30)得分接近低限。212例被试的PDI-Ⅵ和NEO-PI-R得分的多元相关分析显示,反社会型与顺同性负相关(r=-0.25,P<0.01),边缘型与神经质正相关(r=0.32,P<0.01),与顺同性和严谨性负相关(r=-0.22,P<0.01;r=-0.22,P<0.01),表演型与顺同性和严谨性负相关,(r=-0.16,P<0.05;r=-0.15,P<0.05),自恋型与顺同性负相关(r=-0.31,P<0.01)。结论:B群人格障碍患者的五因素人格特征较正常成年人有较大偏差,有较多的负性情绪体验,冲动,人际关系冷漠,缺乏自信等。本研究中B群人格障碍的五因素特征不典型。     

GMP法玻璃化冷冻人胚胎的效果观察

目的探索GMP法玻璃化冷人胚胎的可行性。方法选用IVF中多精受精的4-10细胞阶段的胚胎为研究对象，先后在玻璃化1液（10％乙二醇＋10％二甲亚砜）和玻璃化2液（20％乙二醇＋20％二甲亚砜＋0．3M蔗糖）中分别平衡1mim和0．5mim，然后用GMP管虹吸胚胎直接浸入液氮。其结果与常规的程序化慢冻进行比较。结果GMP法的存活率、胚胎完整率、卵裂球存活率比程序化慢冻法更高（分别为94．4％对73．9％，P〈0．01；79．6％对33．9％，P〈0．01；91．6％对67．8％；P〈0．01），但卵裂球完全死亡率更低（0％对10、4％；P〈0．01）。结论GMP法玻璃化冷冻人胚胎是可行的。     

Enhanced responses of the anterior cingulate cortex neurones to colonic distension in viscerally hypersensitive rats

The anterior cingulate cortex (ACC) is critically involved in processing the affective component of pain sensation. Visceral hypersensitivity is a characteristic of irritable bowel syndrome. Electrophysiological activity of the ACC with regard to visceral sensitization has not been characterized. Single ACC neuronal activities in response to colorectal distension (CRD) were recorded in control, sham-treated rats and viscerally hypersensitive (EA) rats (induced by chicken egg albumin injection, i.p ). The ACC neurones of controls failed to respond to 10 or 30 mmHg CRD; only 22% were activated by 50 mmHg CRD. Among the latter, 16.4% exhibited an excitatory response to CRD and were labelled 'CRD-excited' neurones. In contrast, CRD (10, 30 and 50 mmHg) markedly increased ACC neuronal responses of EA rats (10%, 28% and 47%, respectively). CRD produced greater pressure-dependent increases in ACC spike firing rates in EA rats compared with controls. Splanchnicectomy combined with pelvic nerve section abolished ACC responses to CRD in EA rats. Spontaneous activity in CRD-excited ACC neurones was significantly higher in EA rats than in controls. CRD-excited ACC neurones in control and EA rats (7 of 16 (42%) and 8 of 20 (40%), respectively) were activated by transcutaneous electrical and thermal stimuli. However, ACC neuronal activity evoked by noxious cutaneous stimuli did not change significantly in EA rats. This study identifies CRD-responsive neurones in the ACC and establishes for the first time that persistence of a heightened visceral afferent nociceptive input to the ACC induces ACC sensitization, characterized by increased spontaneous activity of CRD-excited neurones, decreased CRD pressure threshold, and increased response magnitude. Enhanced ACC nociceptive transmission in viscerally hypersensitive rats is restricted to visceral afferent input.     

流行性乙型脑炎病毒活疫苗株SA14-14-2基因稳定性研究

目的 通过研究流行性乙型脑炎活疫苗减毒株基因稳定性，从分子水平证实流行性乙型脑炎活疫苗的遗传稳定性。方法分析流行性乙型脑炎活疫苗主种子、工作种子及其相应的疫苗病毒E蛋白基因核苷酸和氨基酸序列，并与其强毒株和基因库中乙脑病毒减毒株(AF15119)比较。结果乙脑活疫苗主种子、工作种子及其相应的疫苗病毒的E蛋白基因核苷酸序列完全相同。这些病毒E蛋白的氨基酸序列与基因库中乙脑病毒弱毒株(AF315119)比较显示第E447位点氨基酸有差异。结论乙脑病毒活疫苗减毒株遗传学特性稳定。     

重组MHCⅠ类分子K^b-EGFP分子的构建及表达

目的 构建绿色荧光蛋白标记的MHCⅠ类分子K^b(K^b-EGFP融合蛋白)，并在哺乳动物细胞中获得功能性表达。方法 RT-PCR的方法获得小鼠MHCⅠ类分子K^b cDNA，克隆入真核表达载体pEGFP-N1中EGFP分子的上游，脂质体转染COS-7细胞，G418加压筛选获得抗性重组细胞。荧光显微镜下观测重组分子的表达及胞内定位情况。结果 分子克隆的方法获得了预期的重组质粒，质粒经DNA序列测定证实阅读框无误，转染COS-7细胞后的抗性克隆胞浆内可以观察到明亮的绿色荧光，特征性地分布于细胞质膜，及核周、膜下的囊泡结构中。结论 所构建的K^b-EGFP融合蛋白在细胞内表达后具有MHCⅠ类分子的特征性分布，提示该重组分子具有野生型MHCⅠ类分子的生物学特性和胞内分布特征，为进一步深入研究MHCⅠ类分子胞内动态分布及分子功能提供了良好的细胞和分子模型。     

EIAV减毒疫苗诱导马外周血单个核细胞Th1型细胞因子的转录

目的：探讨马传染性贫血病毒驴白细胞减毒疫苗免疫马后,外周血单个核细胞中Th1型细胞因子转录水平与免疫保护应答的关系,揭示DLV的免疫保护机制。方法：应用分子克隆及实时定量RT-PCR技术,建立了马PBMC中IFNγ-、IL-2、IL-12 mRNA转录水平的定量检测方法,定期观察4组（疫苗免疫组、阴性对照组、强毒株阳性对照组、自然感染组）12匹马外周血PBMC中细胞因子的转录水平及分布特征,同时监测体温变化等指标。疫苗株免疫动物8个月后,用EIAV强毒株攻击,观察攻击前后细胞因子转录水平的变化。结果：DLV免疫马,在免疫后3周外周血PBMC中IFN-γ、IL-2转录量显著高于阴性对照组及自然感染组（P〈0.01）;免疫后用EIAV强毒株攻击,IFNγ-、IL-2和IL-12转录的量明显升高,免疫马获得完全保护;强毒株攻击阳性对照马IFN-γ、IL-2转录量随疾病进展波动,发热期下降。结论：本研究首次证明EIAV减毒疫苗可诱导马外周血PBMC中IFN-γ、IL-2、IL-12基因高效转录,其转录水平与DLV的免疫保护密切相关,此结果在分子水平为阐明DLV的免疫保护机制提供了新的实验依据。     

强力霉素影响基质金属蛋白酶活性和血管重构的实验研究

目的：观察强力霉素对损伤动脉组织中基质金属蛋白酶（MMP）活性的抑制作用，并探讨强力霉素对血管平滑肌细胞增殖、动脉内膜增生、管腔重构的影响。方法:球囊导管扩张动脉的方法建立大鼠颈总动脉损伤模型。治疗组用强力霉素30 mg·kg^-1·d^-1干预。明胶酶谱法测定损伤动脉组织中MMPs的活性。用HE染色、VVG染色、免疫组化标记α-actin和增殖细胞核抗原的方法观察损伤动脉内膜厚度、管腔重构及平滑肌细胞增殖的情况。结果:①强力霉素治疗组MMP-9活性在术后24 h、3 d分别比对照组低26.3％、34.5％（P<0.01）；MMP-2活性在术后7 d比对照组低40.0％（P<0.01）。②强力霉素治疗使术后7 d内膜平滑肌细胞增殖率（43.23％±1.06％）显著低于对照组（62.76％±1.02％）（P<0.01）；使术后14 d、28 d新生内膜厚度比对照组分别少32.0％、38.8％（P<0.01），而管腔面积比对照组多58.0％、90.4％（P<0.01） 。结论：强力霉素可以显著降低血管损伤后MMPs活性，抑制内膜平滑肌细胞的增殖、新生内膜增生以及管腔重构，提示它可能具有防治PTCA术后再狭窄的作用。     

多房性囊性肾细胞癌的临床病理学特征和分子遗传学观察

目的 探讨多房性囊性肾细胞癌(MCRCC)的临床病理及分子遗传学改变特征.方法 对1990年1月至2006年12月间第二军医大学附属长海医院病理科病理诊断为肾细胞癌(706例)中的21例MCRCC进行了光镜、免疫表型观察和微卫星不稳定性(MSI)及杂合性缺失(LOH)检测.结果 21例MCRCC患者年龄34～72岁(平均年龄50岁),男19例,女2例.肉眼或镜下血尿3例,18例因体检而偶然发现.10例肿瘤位于左肾,11例位于右肾.T1期18例,T2期2例,T3期1例.其中1例侵犯肾包膜,均无淋巴结转移.随访20例,无一例死于肾癌,未发现肾癌转移或复发病例.组织学观察,肿块最大直径0.3～10.0 cm(平均3.6 cm).切面呈多房性囊性,囊腔大小不等,囊腔间隔厚薄不均(0.1～0.5 cm.平均0.2 cm),内含淡黄色液体或胶冻样或血性液体.囊腔内衬单层或多层上皮细胞,胞质透明或淡嗜酸性.囊腔间隔内也可见呈巢状或腺泡状排列的透明细胞.Fuhrman 1级16例,Fuhrman 2级5例.免疫组织化学观察,瘤细胞波形蛋白、ABC、CAM5.2、上皮细胞膜抗原(EMA)均呈阳性表达,CD10阳性表达6例.神经元特异性烯醇化酶(NSE)阳性表达16例.分子遗传学观察,21例标本中,PCR扩增成功的有11例,5例发现微卫星变异,3例有LOH,其中D3S1560基因座2例,D14S617基因座1例;2例有MSI,分别为D9S168和D14S617各1例.结论 MCRCC是一种少见的肾囊性肿瘤,具有独特的临床和病理学特征,肿瘤预后良好,可通过手术切除而治愈.     

Bb2Bb3 regulation of murine Lyme arthritis is distinct from Ncf1 and independent of the phagocyte nicotinamide adenine dinucleotide phosphate oxidase

Several quantitative trait loci regulating murine Lyme arthritis severity have been mapped, including a highly significant linkage found on chromosome 5, termed Bb2Bb3. Within this region, the Ncf1 gene of the phagocyte nicotinamide adenine dinucleotide phosphate (NADPH) oxidase has recently been identified as a major regulator of arthritis severity in rodent models of rheumatoid arthritis, an effect attributed to protective properties of reactive oxygen species. To assess the role of Ncf1 in Lyme arthritis, we introgressed Bb2Bb3 from severely arthritic C3H/He mice onto mildly arthritic C57BL/6 mice. This increased Lyme arthritis severity, whereas the reciprocal transfer conferred protection from disease. A single nucleotide polymorphism was identified in the Ncf1 gene that did not influence the protein sequence or expression of Ncf1. Although polymorphonuclear leukocytes from C57BL/6 mice generated a greater oxidative burst than polymorphonuclear leukocytes from C3H/He mice, studies with the Bb2Bb3 congenic mice demonstrated this difference was not linked to Ncf1 alleles. Furthermore, Lyme arthritis severity was not altered in mice lacking either the Ncf1 or Gp91phox subunits of the NADPH oxidase complex. Together, these results argue that Ncf1 is not a candidate gene for regulation of Lyme arthritis and reveal Lyme arthritis to be independent of NADPH oxidase activity, distinguishing it from other models of rheumatoid arthritis.