人子宫内膜基质细胞和腺体细胞的分离培养及鉴定

目的 人子宫内膜分离培养并得到纯度较高的内膜基质细胞和腺体细胞。 方法 采用二次滤网过滤法进行分离并通过光镜观察和免疫细胞化学染色对其进行鉴别。 结果 基质细胞在接种后 0 5h开始贴壁 ,可见梭形和多角形两种形态的细胞。免疫细胞化学染色显示这两种细胞均具有波形蛋白免疫反应性 ,反应率可达95 %以上 ,而细胞角蛋白无免疫反应性 ,提示它们为来源于内膜基质的基质细胞。大部分腺体细胞在接种 2 4h后贴壁 ,培养后 4d左右腺体细胞呈旋涡状排列 ,单个细胞为多角形 ,核大而圆。腺细胞呈细胞角蛋白免疫反应性 ,反应率在 90 %以上。

结合密集连接块和自注意力机制的腺体细胞分割方法

针对目前常用的细胞分割方法在对腺体细胞进行分割时易出现误分割和分割不精细的问题,提出一种以U-Net为基本框架,结合密集连接块和自注意力机制的腺体细胞分割模型.首先将U-Net结构中卷积层组合构建成密集连接块,以不同尺度从图像中提取信息;然后在解码端引入自注意力机制,通过对局部特征建立丰富的上下文依赖模型,抑制不必要的特征传播,提高腺体细胞分割的精度.在2015MICCAI腺体分割挑战赛数据集上的实验结果表明,与U-Net等其他模型相比,在增加少量参数的情况下,该模型在F1值、MeanDice和Hausdorff距离评价指标上均具有较大的提升.

胃溃疡旁腺体细胞增殖动力和粘液分泌状况的实验观察

为探讨胃癌发病与胃溃疡之间的关系,我们对大白鼠实验性胃溃疡进行组织病理学、~3H—TdR标记和粘液组化染色,观察了胃溃疡旁粘膜腺体细胞的增殖动力学变化和粘液分泌状况,结果表明:大鼠胃粘膜形成溃疡之后溃疡旁胃腺细胞增生十分活跃,其机能和结构呈现各种异常。胃粘膜在机能上的肠化现象可先于形态学变化。

三维培养的鼻粘膜腺体细胞中粘蛋白基因表达

粘液过量分泌是慢性呼吸道疾病的主要特征之一,慢性鼻窦炎的鼻粘液主要来源于鼻腔及鼻窦的粘膜下腺体细胞,而粘蛋白是鼻粘液的主要成分。目的:研究培养的鼻粘膜腺体细胞中粘蛋白基因的表达。方法:采用逆转录PCR(RT-PCR)技术,检测粘蛋白基因MUC2,MUC5AC,MUC5B的表达。结果:三维培养在Ⅰ型胶原蛋白凝胶内的人鼻粘膜腺体细胞(HNG)中,MUC2,MUC5AC和MUC5B均有表达,在培养液中加入大肠杆菌内毒素(LPS)8小时(100ng/ml)后,MUC2,MUC5AC表达明显增强,而MUC5B表达无变化。结论:实验结果提示,体外培养的HNG细胞可以作为探讨慢性呼吸道疾病粘液过量分泌的研究模型,LPS可以促进部分粘蛋白基因的表达。

基于改进U-net网络的腺体细胞图像分割算法应用研究

针对医学领域中腺体细胞分割问题,提出基于改进U-net网络的腺体细胞图像分割算法,提供一种高可用性图像处理模型。该模型能够增强腺体细胞特征,减少信息丢失,借用OpenCV对腺体细胞轮廓进行颜色处理。采用U-net网络结合空洞残差模块,提高细胞分割精确度。文章方法在ISBI数据集评测,设计多组实验对比,验证可行性,实验结果Dice系数达0.925,表明对腺体细胞图像分割算法存在较高应用价值。

宫颈管不典型腺体细胞临床意义探讨

目的 探讨宫颈管中不典型腺体细胞 (AGCUS)的临床意义。方法 1998年 3月至 2 0 0 1年 8月对门诊16 6 4例患者采用瑞典欧立宫毛刷法施行宫颈管细胞学筛查 ,117例宫颈管细胞中出现不典型腺体细胞 (占7 0 % ) ,为明确此种细胞的性质及来源 ,对AGCUS患者进行子宫分段诊刮及组织病理学检查。结果 病理学检查为正常子宫内膜组织 6 1例 (占 5 2 1% ) ,子宫内膜良性病变 35例 (占 2 9 9% ) ,子宫内膜不典增生 14例 (占11 9% ) ,子宫内膜腺癌 7例 (占 5 9% )。 14例子宫内膜不典型增生中 ,3例行手术治疗 ,术后有 1例诊断为子宫内膜腺癌。结论 不典型腺体细胞在宫颈管细胞学中出现率较低 ,但常是子宫严重疾病的标志 ,对这类患者应作进一步的检查和跟踪随访。

胃肠道细胞生物学的中西医结合研究进展

近年来,消化系统疾病在中西医结合的基础和临床研究中取得长足进步,就基础研究而言,细胞生物学的研究,值得关注.本文就国内外对胃肠道细胞生物学的中西医结合研究进展作一综述.

子宫腺肌病的经阴道超声诊断

子宫腺肌病即子宫肌层内子宫内膜异位症，是由具有功能的子宫内膜腺体细胞及间质细胞向肌层侵蚀，伴随子宫平滑肌细胞增生而引起的一种良性病变。其临床诊断较困难，常规腹部超声检查时，由于盆腔脏器位置深，探头频率低，且受患者腹壁脂肪厚度、膀胱充盈程度及子宫位置等影响，声像图往往显示不满意。

干燥综合征患者泌尿系统阳性超声表现探讨

干燥综合征(Sj觟gren’s syndrome,SS)是一种累及外分泌腺体的慢性炎症性自身免疫病,主要表现为淋巴细胞和浆细胞对外分泌腺体的进行性侵犯,使腺体细胞破坏,功能丧失。由于泪腺、涎腺的受累,临床多表现为口眼干燥,此外,尚有其它外分泌腺及腺体外其它器官的受累而出现多系统损害症状。泌尿系统是常见受累器官之一,但有关泌尿系统损害的超声声像图研究甚少。

埃及用禽粪加VC和鲜酵母养殖罗非鱼

埃及水产科研人员用禽粪添加生素C(VC)和鲜酵母进行养殖尼罗罗非鱼试验。试验饲料中禽粪所占比例分为0％、10％、20％,30％、40％,添加维生素C3g/kg,添加鲜酵母10％。试验用尼罗罗非鱼平均体重42.44g,体长13.24cm,共210尾。投饵试验12周。结果表明。

Characterization of Two Human Lung Adenocarcinoma Cell Lines by Reciprocal Chromosome Painting

Lung cancer is a leading cause of cancer death worldwide. Some lung cancer patients correlate with a gas of radon besides smoking. To search for common chromosomal aberrations in lung cancer cell lines established from patients induced by different factors, a combined approach of chromosome sorting, forward and reverse chromosome painting was used to characterize karyotypes of two lung adenocarcinoma cell lines： A549 and GLC-82 with the latter line derived from a patient who has suffered long-term exposure to environmental radon gas pollution. The chromosome painting results revealed that complex chromosomal rearrangements occurred in these two lung adenocarcinoma cell lines. Thirteen and twenty-four abnormal chromosomes were identified An A549 and GLC-82 cell lines, respectively. Almost half of abnormal chromosomes in these two cell lines were formed by non-reciprocal translocations, the others were derived from deletions and duplication/or amplification in some chromosomal regions. Furthermore, two apparently common breakpoints, HSA8q24 and 12q14 were found in these two lung cancer cell lines.

Persistence of side population cells with high drug efflux capacity in pancreatic cancer

AIM: To investigate the persistence of side population (SP) cells in pancreatic cancer and their role and mechanism in the drug resistance. METHODS: The presentation of side population cells in pancreatic cancer cell line PANC-1 and its proportion change when cultured with Gemcitabine, was detected by Hoechst 33342 staining and FACS analysis. The expression of ABCB1 and ABCG2 was detected by real- time PCR in either SP cells or non-SP cells. RESULTS: SP cells do exist in PANC-1, with a median of 3.3% and a range of 2.1-8.7%. After cultured with Gemcitabine for 3 d, the proportion of SP cells increased significantly (3.8% ± 1.9%, 10.7% ± 3.7%, t = 4.616, P = 0.001 < 0.05). ABCB1 and ABCG2 expressed at higher concentrations in SP as compared with non-SP cells (ABCB1: 1.15 ± 0.72, 5.82 ± 1.16, t = 10.839, P = 0.000 < 0.05; ABCG2: 1.16 ± 0.75, 5.48 ± 0.94, t = 11.305, P = 0.000 < 0.05), which may contribute to the efflux of fluorescent staining and drug resistance. CONCLUSION: SP cells with inherently high resistance to chemotherapeutic agents do exist in pancreatic cancers, which may be candidate cancer stem cells contributing to the relapse of the tumor.

Effects of angiotensin Ⅱ receptor antagonist, Losartan on the apoptosis, proliferation and migration of the human pancreatic stellate cells

AIM： To investigate the effects of AT, （Type 1 angiotensin Ⅱ receptor） antagonist （Losartan） on the apoptosis, proliferation and migration of the human pancreatic stellate cells （hPSCs）. METHODS： hPSCs were isolated from pancreatic sample of patients with pancreatic carcinoma using radioimmunoassay （RIA） technique to detect the concentration of AngⅡ in culture media and cell homogenate. Immunocytochemistry （ICC） and in situ hybridization （ISH） methods were utilized to test AT1 expression in hPSCs. Effects of Losartan on hPSCs proliferation, apoptosis and migration were investigated using BrdU incorporation, TUNEL, flow cytometry （FCM）, and phase-contrast microscope separately when cells treated with Losartan. Immunofluorescence and Western blot were applied to quantify the expression of type Ⅰ collagen in hPSCs. RESULTS： There exists AT1 expression in hPSCs, while no AngⅡ was detected in culture media and cell homogenate. Losartan induces cell apoptosis in a doseand time-dependent manner （apparently at 10^-5 mol/L）, no pro-proliferative effect was observed in the same condition. Corresponding dosage of Losartan can also alleviate the motion capability and type Ⅰ collagen content of hPSCs compared with AngⅡ treatment and non-treatment control groups. CONCLUSION： These findings suggest that paracrine not autocrine functions of AngⅡ may have effects on hPSCs, which was mediated by AT1 expressed on cells, while Losartan may exert anti-fibrotic effects by inhibiting hPSCs motion and partly by inducing apoptosis.

Mechanism for Src activation by the CCK2 receptor:Patho-physiological functions of this receptor in pancreas

AIM： TO investigate in vivo, whether CCK2 receptors （CCK2R） regulate proteins known to play a crucial role in cell proliferation and cancer development and analyse in vitro the molecular mechanisms that lead to Src activation; in particular, to identify the domains within the CCK2R sequence that are implicated in this activation. METHODS： The expression and activation of Src and ERK were studied in vivo using immunofluorescence and western-blot techniques. We used pancreatic tissues derived from wild type or Elas-CCK2 mice that expressed the CCK2R in pancreatic acini, displayed an increased pancreatic growth and developed preneoplastic lesions. The pancreatic tumor cell line AR4-2J expressing the endogenous CCK2R or COS-7 cells transiently transfected with wild type or mutant CCK2R were used as in vitro models to study the mechanism of Src activation. Src activation was measured by in vitro kinase assays, ERK activation by western blot using antiphospho-ERK antibodies and the involvement of Src in gastrin-induced cell proliferation by MTT test. RESULTS： We showed in vivo that the targeted CCK2R expression in the pancreas of Elas-CCK2 mice, led to the activation of Src and the ERK pathway. Src was activated upstream of the ERK pathway by the CCK2R in pancreatic tumoral cells and contributed to the proliferative effects mediated by this receptor. In vitro results demonstrated that activation of the Src/ERK pathway by the CCK2R required the NPXXY motif, located within the CCK2R sequence at the end of the 7^th transmembrane domain, and suggested the putative role of Gq in this mechanism. CONCLUSION： Deregulation of the Src/ERK pathway by the CCK2R might represent an early step that contributes to cell proliferation, formation of preneoplastic lesions and pancreatic tumor development.

Thyroid hormone regulation of apoptotic tissue remodeling during anuran metamorphosis

Anuran metamorphosis involves systematic transformations of individual organs in a thyroid hormone (TH)-dependent manner. Morphological and cellular studies have shown that the removal of larval or- gans/tissues such the tail and the tadpole intestinal epithelium is through programmed cell death or apop- tosis. Recent molecular investigations suggest that TH regulates metamorphosis by regulating target gene expression through thyroid hormone receptors (TRs), which are DNA-binding transcription factors. Cloning and characterization of TH response genes show that diverse groups of early response genes are induced by TH. The products of these TH response genes are believed to directly or indirectly affect the expression and/or functions of cell death genes, which are conserved at both sequence and function levels in different animal species. A major challenge for future research lies at determining the signaling pathways leading to the activation of apoptotic processes and whether different death genes are involved in the regulation of apoptosis in different tissues/organs to effect tissue-specific transformations.

c-Fos enhances the survival of thymocytes during positive selection by upregulating Bcl-2

T cells are derived from progenitor thymocytes, of which only a minority receive the appropriate TCR signal, undergo positive selection and mature. Owing to the very short lifespan of thymocytes, the prerequisite for posi- tive selection is survival. TCR signal-induced Bcl-2 expression is believed to play a dominant role in the survival of positively selecting thymocytes, but how Bcl-2 is directly regulated is unknown. Here we report that the immediate early gene （IEG） c-Fos can stimulate the expression of Bcl-2, depending on a specific AP-l-binding site in the Bcl-2 promoter. In c-Fos transgenic （Fos-Tg） mice, c-Fos binds to this site and promotes the expression of Bcl-2. As a result, Fos-Tg thymocytes exhibited enhanced survival, and more mature single-positive （SP） thymocytes were generated, even on a unique TCR background. The TCR repertoire remained normal in Fos-Tg mice. Our results identified c-Fos as the mediator of the stimulatory effect of TCR signaling on Bcl-2 expression. Therefore, c-Fos, as an IEG, because of its early response ability, can quickly rescue the survival of short-lived thymocytes during positive selection. Our results provide novel insight into the mechanism regulating the survival of positively selecting thymocytes.