人体宫颈上皮细胞的体外培养及鉴定

随着宫颈癌及癌前期疾病发病率的增加,人们对其发病机制的的研究也越来越深入。自对人宫颈癌细胞体外培养获得成功后,学者对不同类型的宫颈细胞的体外培养实验也相继展开研究。通过对宫颈上皮细胞特定角蛋白及免疫标记物的测定来鉴定是否为宫颈细胞,通过观察培养出的各类型宫颈细胞的生物学特性是进一步研究宫颈疾病发生发展规律从而找出治疗宫颈疾病的最佳途径。

激光共焦显微拉曼光谱技术在人舌鳞癌细胞检测中的应用

目的:采用激光共焦显微拉曼光谱技术对人舌鳞癌细胞CAL-27进行检测,并将其与正常人体口腔黏膜上皮细胞(NHOK)进行对比和分析,从而从分子层面区分CAL-27和NHOK。方法:培养CAL-27和NHOK,并用激光共焦显微拉曼光谱对其进行检测,以及采用主成分分析法和线性判别技术对其进行分析。结果:对比CAL-27和NHOK光谱差谱,发现谱宽于735、959、1 334、1 575 cm-1处,CAL-27的峰值比NHOK强,但在1 033 cm-1处,则相对要弱。结合主成分分析和线性判别技术的灵敏度为96.8%,特异性为90.3%,诊断率为93.5%。受试者操作特性曲线下面积为0.971。结论:采用激光共焦显微拉曼技术可区分CAL-27和NHOK。

亚细胞尺度氚非均匀分布下单粒子效应的蒙特卡罗模拟计算

利用蒙特卡罗程序Geant4构建了正常肺上皮细胞体素模型,结合Geant4自建DNA模型,模拟计算了氚在亚细胞尺度不同靶区的物理作用和DNA损伤效应,分析了氚处于不同滞留状态下β单粒子对细胞内环境及DNA的影响。结果表明,氚的滞留状态对不同靶区的作用呈规律性变化,源项为细胞核时,源靶组合N->N与N->C的能量沉积E_(细胞核)/E_(细胞质)比值差异最为显著,微剂量学特征值SN->N最大。β单粒子作用于DNA产生的集簇损伤尺寸一般<3,且多为简单单链断裂(SSSB)类型,尽管产生的集簇损伤尺寸≥3的比例(<5%)很低,但仍有产生集簇损伤尺寸高达6的可能。源项为细胞核时,产生双链断裂(DSB)的产额是单链断裂(SSB)的5%,其中复杂双链断裂(CDSB)约占DSB的20%,相比细胞质、细胞源项,CDSB产额最大。

Parthenolide protects human lens epithelial cells from oxidative stress-induced apoptosis via inhibition of activation of caspase-3 and caspase-9

The apoptosis of lens epithehal cells has been proposed as the common basis of cataract formation, with oxidative stress as the major cause. This study was performed to investigate the protective effect of the herbal constituent parthenolide against oxidative stress-induced apoptosis of human lens epithelial （HLE） cells and the possible molecular mechanisms involved. HLE cells （SRA01-04） were incubated with 50 μM H2O2 in the absence or presence of different doses of parthenolide （10, 20 and 50 μM）. To study apoptosis, the cells were assessed by morphologic examination and Annexin V-propidium iodide double staining flow cytometry; to investigate the underlying molecular mechanisms, the expression of caspase-3 and caspase-9 were assayed by Western blot and quantitative RT-PCR, and the activities of caspase-3 and caspase-9 were measured by a Chemicon caspase colorimetric activity assay kit. Stimulated with H202 for 18 h, a high fraction of riLE cells underwent apoptosis, while in the presence ofparthenolide of different concentrations, dose-dependent blocking of HLE cell apoptosis was observed. The expression of caspase-3 and caspase-9 induced by H202 in HLE cells was significantly reduced by parthenolide both at the protein and mRNA levels, and the activation ofcaspase-3 and caspase-9 was also suppressed by parthenolide in a dose-dependent manner. In conclusion, parthenolide prevents HLE cells from oxidative stress-induced apoptosis through inhibition of the activation ofcaspase-3 and caspase-9, suggesting a potential protective effect against cataract formation.

Effects of recombinant human growth hormone on enterocutaneous fistula patients

AIM： To explore the effects of recombinant human growth hormone （rhGH） on intestinal mucosal epithelial cell proliferation and nutritional status in patients with enterocutaneous fistula. METHODS： Eight patients with enterocutaneous fistulas received recombinant human growth hormone （10 ug/d） for 7 d. Image analysis and immunohistochemical techniques were used to analyse the expression of proliferating cell nuclear antigen （PCNA） in intestinal mucosal epithelial cells in biopsy samples from the patients who had undergone an endoscopic biopsy through the fistula at day 0, 4 and 7. Body weights, nitrogen excretion, serum levels of total proteins, albumin, prealbumin, transferrin and fibronectin were measured at day 0, 4 and 7. RESULTS： Significant improvements occurred in the expression of PCNA in the intestinal mucosal epithelial cells at day 4 and 7 compared to day 0 （24.93 ± 3.41%, 30.46 ± 5.24% vs 12.92 ± 4.20%, p 〈 0.01）. These changes were accompanied by the significant improvement of villus height （500.54 ± 53.79 um, 459.03 ± 88.98um vs 210.94 ± 49.16 um, P 〈 0.01）, serum levels of total proteins （70.52 ± 5.13 g/L, 74.89 ± 5.16 g/L vs 63.51 ± 2.47 g/L, P 〈 0.01）, albumin （39.44 ± 1.18 g/L, 42.39 ± 1.68 g/L vs 35.74 ± 1.75 g/L, P 〈 0.01） and fibronectin （236.3 4- 16.5 mg/L, 275.8± 16.9 mg/L vs 172.5 ± 21.4 mg/L, P 〈 0.01） at day 4 and 7, and prealbumin （286.38 ± 65.61 mg/L vs 180.88 ± 48.28 mg/L, P 〈 0.05）, transferrin （2.61 ± 0.12 g/L vs 2.41 ±0.14 g/L, P 〈 0.05） at day 7. Nitrogen excretion was significantly decreased at day 7 （3.40 ± 1.65 g/d vs 7.25 ± 3.92 g/d, P 〈 0.05）. No change was observed in the body weight. CONCLUSION： Recombinant human growth hormone could promote intestinal mucosal epithelial cell proliferation and protein synthesis in patients with enterocutaneous fistula.

基于柴胡皂苷抗溃疡药效成分的药动药效相关性

目的:明确柴胡皂苷发挥治疗胃溃疡作用的化学成分,开展柴胡皂苷治疗胃溃疡药动药效相关性研究,为探讨其药效物质及作用机制提供前期实验基础。方法:采用高分辨质谱,一、二级质谱解析方式解析柴胡皂苷化学成分,同时取SD大鼠灌胃柴胡皂苷提取液(300 mg·kg^(-1))后不同时间点采血,采用高效液相色谱法对其入血成分柴胡皂苷B_2,柴胡皂苷D进行血药浓度测定;分别对柴胡皂苷B_2,柴胡皂苷D血药浓度与人体胃黏膜上皮细胞体外增殖率,超氧化物歧化酶(SOD),丙二醛(MDA)进行时效相关性分析,评价柴胡皂苷作用特点及可能机制。结果:柴胡皂苷抗溃疡化学成分主要为柴胡皂苷A,D,B_2,I,E,prosaikogenin D,F,G;柴胡皂苷含药血浆与损伤的人体胃黏膜上皮细胞增殖率具有良好的相关性,其吸收也与血中MDA,SOD水平呈一定的时效关系。结论:该研究明确了柴胡皂苷发挥治疗胃溃疡作用的化学成分,探讨了入血成分与药效相关性,其作用机制可能与MDA,SOD蛋白调控有关。

Ureaplasma urealyticum-derived lipid-associated membrane proteins introduce IL-6, IL-8, and TNF-α cytokines into human amniotic epithelial cells via Toll-like receptor 2

Objective： The purpose of this study was to determine the role of Ureaplasma urealyticum-derived lipidassociated membrane proteins （LAMPs） in the host innate immune system, specifically their effect on Toll-like receptors （TLRs）. Methods： LAMPs were derived from U. urea/yticum strains, and human amniotic epithelial cells （HAECs） were isolated from healthy full-term placentas. Cytokine concentrations were determined by enzyme-linked immunosorbent assay （ELISA） and TLR2 mRNA by real-time PCR. Expression of TLR2 was confirmed by Western blotting and immunohistochemistry. Results： LAMPs induced HAECs to produce inflammatory cytokines interleukin （IL）-6, IL-8, and tumor necrosis factor （TNF）-α. Cytokine production was reduced after blocking TLR2 using TLR2 inhibitor （anti-hTLR2-IgA）. Conclusions： LAMPs isolated from U. urealyticum induced TLR2-dependent up-regulation of inflammatory genes and cytokines in HAECs.