单细胞显微切割术在霍奇金淋巴瘤中的应用

目的 :应用单细胞显微切割技术分离和研究霍奇金淋巴瘤 (Hodgkin slymphoma ,HL)瘤细胞 ,并检测其是否存在人巨细胞病毒 (humancytomegalovirus,HCMV)感染。 方法 :采用单细胞显微切割技术从HL组织切片中获得其瘤细胞Reed Stern berg/Variant (RS/V)细胞 ,提取DNA后 ,利用PCR扩增HCMV立即早期基因片段。结果 :单细胞显微切割可将RS/V细胞从HL淋巴组织中独立分离出来 ,收集切割的瘤细胞可获得用于PCR扩增的模板DNA ,在 13例HL石蜡组织切片中有 4例经PCR扩增出 149bp的目的片段。 结论 :单细胞显微切割技术适用于HL瘤细胞研究 ;

人类染色体显微切割技术及其应用

人类染色体显微切割技术可从特定区域切取所需的染色体片段，结合PCR、分子克隆、DNA测序和FISH等方法，用于染色体特定区、带的DNA文库构建、特异性探针的制备、疾病遗传学特征的分析和有关基因的鉴定、分离、克隆和定位等。而且，该技术有助于人类基因组物理图谱的绘制。

基于多模块组合的生物显微图像降噪方法研究

在利用细胞显微切割术进行显微成像时,由于存在仪器热噪声、样本表面特性和样本染色制备情况等因素干扰,显微成像质量容易受损。针对这个问题,提出一种基于多模块组合的去噪网络模型(DNMMC),该网络模型使用通道关联模块、多尺度去噪模块和融合压缩模块对输入图像进行处理并在最后输出降噪图像。将所提方法与目前主流的去噪方法在定量测量和视觉质量感知方面进行比较,实验结果表明DNMMC在对生物显微图像降噪上具有更高的鲁棒性。可见,该方法可以显著提高生物显微镜的成像质量。

Laser Capture Microdissection Combined with RNA in vitro Linear Amplification Detecting the Relevant Genes of Bladder Cancer

Objective： Laser capture microdisection has become indispensable to the analysis of the difference of gene expression between human bladder transitional cell and bladder transitional cell carcinoma （BTCC）. However, to obtain sufficient RNA from laser-capture microdissected cells is quite difficult. The study was designed to determinc a feasible technical routine to isolate transitional cells from bladder membrane, separate carcinoma cclls from stromal cells and to amplify the RNA isolated from laser-capture microdissected cells. Methods： Bladder transitional cell were obtained from frozen sections of bladder membrane applying LCM, by the same token, BTCC cells from frozen sections of BTCC tissue. Then RNA was extracted and linearly amplified in vitro. The expression levels of β-actin in primary total RNA and amplified RNA were detected using RT-PCR. Results： That RNA integrity was good after LCM was confirmed by control experiment Ⅰ; By control experiment Ⅱ, the correlation between the number of LCM-shooting and RNA quantity undcr arranged conditions was preliminarily confirmed. About 0.5-2.5kb RNA fragments were obtained after RNA amplification and β-actin levels were integral. Conclusion： Laser capture microdissection combined with RNA linear amplification in vitro can be successfully applied to obtain pure objective cells for research. The integrity of the amplified RNA is good and can be employed in further research.

重组人角质细胞生长因子异构体抑制兔角膜基质细胞增殖分化的研究

目的观察重组人角质细胞生长因子异构体(recombinant human keratinocyte growth factor variant,K102)对实验性兔角膜损伤的治疗作用。方法108只家兔均在显微镜下造成板层切割损伤模型,并分成K102药物大剂量(0.5 mg/ml)、中剂量(0.25 mg/ml)、小剂量(0.125 mg/ml)治疗组、贝复舒对照组(贝复舒滴眼液治疗,2 400 AU/ml)、地塞米松对照组(地塞米松磷酸钠滴眼液治疗,0.25 mg/ml)和空白组(K102滴眼液的溶媒滴眼治疗)。通过裂隙灯观察角膜上皮下雾状浑浊(Haze)差别。于术后30、90 d处死动物取角膜行光镜及电镜检查。结果术后30、90 d,K102药物治疗的各组Haze评分及前50μm角膜基质层内平均角膜细胞数均明显低于阳性对照组及空白组(P<0.05),光镜及电镜检查结果显示K102大剂量组角膜成纤维细胞活化及增殖程度明显轻于贝复舒对照组、空白组。结论K102滴眼液可抑制角膜损伤后的角膜基质细胞过度增殖、活化,减少早期Haze形成。