羟基喜树碱对人胆管癌QBC939裸鼠移植瘤PS2、COX-2的影响

目的探讨羟基喜树碱对人胆管癌QBC939裸鼠移植瘤的乳癌相关肽(PS2)、环氧合酶-2(COX-2)的影响,分析PS2、COX-2与肿瘤发生、发展的关系。方法建立人胆管癌QBC939细胞株裸鼠皮下移植瘤模型,将30只裸鼠随机分成对照组、羟基喜树碱低剂量组和羟基喜树碱高剂量组,观察移植瘤体生长情况,RT-PCR法检测瘤体内PS2、COX-2 mRNA表达。结果与对照组比,羟基喜树碱低、高剂量组都可使移植瘤体的PS2 mRNA表达增高;COX-2 mRNA表达降低,且瘤体生长减慢,与对照组比较差异有统计学意义(P<0.01)。结论羟基喜树碱可降低移植瘤COX-2 mRNA表达,增高移植瘤体的PS2 mRNA表达,并使人胆管癌裸鼠移植瘤生长减慢。     

降钙素基因相关肽与内皮素受体A在兔实验性蛛网膜下腔出血后基底动脉的表达

目的探讨蛛网膜下腔出血(SAH)后脑血管内皮细胞降钙素基因相关肽(CGRP)及内皮素受体A(ETRA)的表达与迟发性脑血管痉挛(DCVS)之间的关系。方法日本大耳白家兔60只,随机分为SAH模型组25只,盐水组25只,穿刺组5只,正常组5只。采用枕大池二次注血法制作SAH模型,灌注处死后取基底动脉制成切片。进行CGRP及ETRA的免疫组化和HE染色,观察CGRP与ETRA的免疫表达和病理结构变化,并用图像分析方法测量血管周长,使用方差分析进行统计学检验。结果基底动脉周长在SAH后均发生明显缩窄,其中在5d时缩小最强烈。CGRP与ETRA免疫阳性标记颗粒在SAH组的基底动脉组织中均有表达,其动态变化规律为CGRP的表达在1h时最明显,在3、5d依次表达减少,5d达到最低点,7d后开始逐渐升高,10d时表达相对比较明显,但未能达到正常水平,CGRP在穿刺组、正常对照组和盐水组也有散在不规则表达。在正常组、穿刺组和盐水组的基底动脉内ETRA有表达;SAH组基底动脉内ETRA在3d组和5d组表现为高表达,以3d组为强,而其中血管内皮细胞表达最强烈,平滑肌也有表达,10d组的表达仍然较强。结论在SAH后脑血管内皮细胞CGRP和ETRA的表达呈现明显的动态改变,但二者的变化趋势并不同步;CGRP和ETRA在DCVS的发生发展中起重要的作用。     

The role of interleukin-8 produced by tumor induced fibroblasts in the development of cutaneous melanoma

Objective To determine the role of interleukin-8 (IL-8) produced by tumor induced fibroblasts in the development of cutaneous melanoma. Methods B16 melanoma cells induced L929 fibroblasts phenotype was transdifferentiated to myofibroblasts (MF) by co-culture in vitro. MF was monitored by morphology and immunophenotype for a-SMA. The level of IL-8 was detected by ELISA. The effect on B16 cell proliferation rate was estimated using MIT method in vitro. Melanoma implanting model was constructed in C57 mice. Results L929 MF phenotype could be modulated by B16 melanoma cells-derived transforming growth factor-β1 (TGF-β1) and elevated the levels of IL-8. L929 MF did not influence the B16 melanoma cells viability in vitro, but shortened the time of tumor formation and increased the incidence rates of tumors in C57 implanting model mice. Conclusion Fibroblasts can be activated by tumor cells and produce IL-8, which acts as an inflammatory cytokine promoting the development of cutaneous melanoma.     

Study on T lymphocyte subsets and NK cells in patients with Graves＇ disease combined with type 2 diabetes

Objective To investigate changes in T lymphocyte subsets and NK cells in patients with simple Graves' disease (GD)and Graves' disease combined with type 2 diabetes mellitus (GD/T2DM). Methods Fifteen cases of GD/ T2DM were selected from our hospital from November 2001 to November 2004. Before and after therapy thyroid function, thyroglobulin antibody (TGA), thyroid microsomal antibody (TMA) and blood glucose level were measured, and T lymphocyte subsets (CD3, CD4, CD8, CD4/CD8) and NK cells (CD56) were measured by immunofluorescence double labeling monoclonal antibody and flow cytometry, respectively. At the same time, comparison was made with simple GD (15 cases), T2DM (15 cases) and healthy control (20 cases). Results Before therapy, CD4/CD8, CD4 and NK cells in GD/T2DM were less than normal, and there was no significant difference in comparison with simple GD (P<0.05). In T2DM group, only CD4/CD8 and CD4 were less than those of healthy controls (P<0.05). When thyroid function recovered after 1 to 3 months of methimazole treatment in both GD/T2DM and simple GD groups, various indexes recovered, which were more obvious in simple GD. Conclusion Immune hypofunction of GD may be the key to the immune abnormality of GD/T2DM, which is more significant than that of simple GD or T2DM. The recovery of thyroid function and immune abnormality is not consistent, and the recovery of GD is more significant than that of GD/T2DM.     

健胃愈疡颗粒对慢性胃炎伴幽门螺杆菌感染胃黏膜PAF、PS2的表达及对上皮细胞凋亡的影响

目的观察健胃愈疡颗粒(JWYY)对幽门螺杆菌相关性胃炎胃黏膜乳癌相关肽(PS2)和血小板活化因子(plate-let activating factor,PAF)表达的变化,并分析其可能的作用机制。方法80例幽门螺杆菌感染慢性胃炎患者随机分为2组,治疗组42例给予健胃愈疡颗粒治疗,对照组38例给予西药三联药物治疗。应用胃镜下活检胃黏膜组织快速尿素酶和Warthin-Starry染色法检测幽门螺杆菌,应用Western blot检测PS2、PAF表达变化。应用末端转移酶介导的dUTP切口末段标记法(TUNEL)检测细胞凋亡。第4周复查胃镜。结果治疗组凋亡指数明显比对照组减少(P<0.05),治疗组总有效率高于对照组(P<0.05)。Western blot检测显示:JWYY颗粒可上调PS2并下调PAF的表达,而对照组对胃黏膜PS2和PAF的表达影响不大。结论健胃愈疡颗粒治疗幽门螺杆菌(Hp)相关性胃炎可能与杀灭Hp、上调PS2和下调PAF的表达来抑制细胞凋亡有关。     

2型糖尿病患者一级亲属正常糖代谢人群胰岛素抵抗分析

目的探讨2型糖尿病(T2DM)患者糖耐量正常的一级亲属胰岛β细胞功能及胰岛素抵抗情况,并分析胰岛素抵抗的相关因素。方法选取T2DM患者糖耐量正常的一级亲属63例,根据胰岛素抵抗指数分为胰岛素抵抗组及非抵抗组;选取30例健康体检者作为正常对照组。对所选的93例对象进行血压、血糖、胰岛素、血脂的测定,并计算稳态模型胰岛素抵抗指数(HOMA-IR)及胰岛β细胞功能指数(HOMA-β)。结果①糖尿病患者一级亲属组与对照组相比,胰岛素水平、HOMA-IR、HOMA-β明显升高(P<0.05);②胰岛素抵抗组体质指数、舒张压、甘油三酯、餐后血糖、HOMA-β明显高于胰岛素非抵抗组(P<0.05);③相关分析示胰岛素抵抗与体质指数、舒张压、甘油三酯、餐后胰岛素呈正相关(P<0.01,P<0.05),与空腹血糖、餐后血糖、空腹胰岛素、收缩压、胆固醇无显著相关性(P>0.05)。结论T2DM患者一级亲属存在胰岛β细胞代偿性的高分泌,早期已经出现胰岛素敏感性的下降和胰岛素抵抗。胰岛素抵抗与体重增加、舒张压升高及甘油三酯的升高有关。提示对糖尿病患者一级亲属应严格控制体重、血压及血脂,对减少T2DM具有积极意义。     

选择性垂体甲状腺激素抵抗综合征患者的临床特点与THRβ基因突变的研究

目的研究2例选择性垂体甲状腺激素抵抗综合征患者的临床特点及甲状腺激素受体β(THRβ)基因突变,探索该病的发病机制。方法收集2例患者及其父母的临床资料并留取外周血标本,提取基因组DNA,应用PCR-DNA直接测序的方法研究患者及父母的THRβ基因有无突变。结果2例患者及父母的THRβ基因第1-10外显子未发现突变。结论散发性垂体甲状腺激素抵抗综合征患者发病可能与THRβ基因突变无关。     

EGFR、Ku70、NF-κB及Bcl-2在Graves病中的表达及意义

目的研究Graves病患者细针穿刺组织中EGFR、Ku70、NF-κB和Bcl-2基因的表达水平,探讨其在Graves病发生发展中的作用。方法 Graves病组患者59例,平均年龄(44.66+12.94)岁,其中男16例,女43例;甲状腺结节周围正常甲状腺组织标本27例为对照组,平均年龄(44.64+14.27)岁,其中男6例,女21例。采用实时荧光定量逆转录-多聚酶链反应(RT-PCR)法检测EGFR、Ku70、NF-κB和Bcl-2在Graves病及对照组甲状腺组织中的表达,t检验分析基因表达水平差异;Pearson相关法分析基因表达与临床特征及相关血清学指标之间的关系。结果上述基因在对照组及Graves病甲状腺组织中都有不同程度的表达。Graves病患者组织中EGFR及Ku70的表达水平明显高于对照组甲状腺组织(1.752±0.660 vs.0.859±0.125,P<0.05;3.304±0.402 vs.0.768±0.102,P<0.001);NF-κB及Bcl-2的表达水平明显低于对照组甲状腺组织(0.578±0.066 vs.0.884±0.085,P<0.001;0.834±0.086vs.1.235±0.261,P<0.05)。Graves病组织中NF-κB与Bcl-2表达水平呈正相关(r=0.399,P<0.001)。Graves病组织中Ku70的表达水平与血清TgAb水平呈正相关(r=0.263,P<0.05),而其他基因表达与相关血清学指标无关(P均>0.05)。结论 Ku70、EGFR、NF-κB、Bcl-2介导的信号转导通路可能参与了Graves病的发生、发展过程。     

姜黄素促进RAW264.7源性M1巨噬细胞向替代激活M2表型极化

目的观察姜黄素对LPS和IFNγ诱导的RAW264.7巨噬细胞(M1)极化的影响及机制。方法不同浓度姜黄素(6.25、12.5、25μmol/L)干预LPS和IFNγ诱导的RAW264.7巨噬细胞(M1)12h,同时再加入20μmol/L GW9662与25μmol/L姜黄素共同干预LPS和IFNγ诱导的RAW264.7巨噬细胞(M1)12h,采用Real-time PCR、ELISA及Western blot方法检测IL-1β、IL-6和M2表型标志分子KLF4、FIZZ1、MGL1、PPARγ的表达,及阻断PPARγ后KLF4和FIZZ1的表达。结果不同浓度姜黄素均能上调LPS和IFNγ诱导的RAW264.7巨噬细胞(M1)的M2标志分子的表达,并且抑制炎症因子IL-1β和IL-6的分泌;阻断PPARγ后,RAW264.7巨噬细胞源性M1表型巨噬细胞表达M2标志分子下调。结论姜黄素通过活化PPARγ促进LPS和IFNγ诱导的RAW264.7巨噬细胞(M1)向M2表型极化。