大鼠肝癌细胞与Ⅰ型胶原的黏附特性及其与细胞周期的关系

肿瘤细胞和胞外基质之间的黏附特性与肿瘤的侵袭转移有密切关系。作者从细胞周期的角度，采用微管吸吮技术和细胞同步技术研究了不同细胞周期肝癌细胞与Ⅰ型胶原裱衬表面的黏附力学特性．结果表明；胸腺嘧啶脱氧核苷、秋水仙碱顺序阻断和胸腺嘧啶脱氧核苷双阻断后释放培养的方法可分别获得G1期和S期肝癌细胞，平均同步率分别为74．09％和90．39％；在研究剂量和时间范围内，肝癌细胞与Ⅰ型胶原的黏附力具有浓度和时间依赖性；S期肝癌细胞和Ⅰ型胶原的黏附力值与G1期和未同步组（对照组）相应值比较明显降低．结果提示；肝癌细胞经血道转移的侵蚀细胞间质阶段，G1期细胞可能起更重要的作用。这一研究对全面认识肝癌的转移机理有重要意义。     

肺癌细胞粘弹特性与超微结构的相关性研究

目的：肺癌是高恶性和高死亡率的肿瘤之一,作者力图从生物力学角度揭示肺 某些生物学特性。材料与方法：本文结合超微结构观察,采用微管吸吮技术定量测定体外培养的低转移人肺腺癌（ＰＡａ）细胞和高转移人肺巨细胞癌（ＰＧ）细胞在长春新碱（ＶＣＲ）作用下的是特性。结果：肺癌细胞粘弹性,在未使用ＶＣＲ前为ＰＡａ细胞大于ＰＧ细胞;随ＶＣＲ的使用和剂量增加,其在ＰＡａ细胞呈浓度依赖性下降,在ＰＧ细胞则表现为低ＶＣＲ浓     

肝细胞粘弹特性及其与细胞骨架结构的相关性研究

用微管吸吮技术研究了正常原代胎肝细胞和肝实质细胞癌细胞的粘弹特性。以此为基础，进一步研究细胞松驰素Ｄ和秋水仙素等两种细胞骨架干扰剂对ＨＦＨ粘弹特性的影响。作者对这一结果进行了简要讨论。     

Ⅲ型前胶原诊断肝硬化和肝癌的临床价值

Ⅲ型前胶原诊断肝硬化和肝癌的临床价值陈泮藻，戴皓洁，王录焕，李振甲肝脏发生纤维化时，纤维增生，产生大量胶原，胶原代谢过程中，形成Ⅲ型前胶原（ＰＣⅢ）或Ⅲ型前胶原肽（ＰⅢＰ）。检测血液中ＰＣⅢ已成为判断肝硬化和肝癌的一个灵敏的指标。本文观察１６１名肝脏...     

人原发性肝癌间质胶原及其mRNA的表达

用ＡＢＣ方法对２９例人肝细胞癌和２例胆管细胞癌组织及癌旁肝细胞Ｉ型胶原、Ⅲ型前胶原进行免疫组化定位。结果显示两种胶原在细胞外分布是一致的。作者将胶原在肝细胞癌中的分布类型分为两大类：血管样分布和非血管样分布。癌组织内胶原呈血管样分布者，肿瘤易浸润生长及门静脉转移，其肝内转移灶发生率也显著高于非血管分布者，预后较差，推测胶原的分布形式可作为判断肝癌患者预后的一个指标，我们还用ｃＤＮＡ－ｍＲＮＡ原位分     

人原发性肝癌Ⅰ型胶原基因的表达

用DNA-m RNA原位分子杂交技术检测了2例正常肝及4例原发性肝癌、癌周肝组织中Ⅰ型胶原基因的表达。结果提示人原发性肝癌组织中肿瘤细胞及间质细胞均可参与Ⅰ型胶原的合成。胶原的合成可能与细胞的活跃增殖有关,其意义尚需进一步探讨。     

整合素beta1亚单位对肝癌细胞与IV型胶原黏附与趋化行为的影响

采用微吸管实验技术 ,测定肝细胞癌 (Hepatocellular carcinom a,HCC)细胞与 IV型胶原裱衬表面的黏附力 ;进一步加入针对整合素 beta1亚单位 (CD2 9)的单克隆抗体 (Anti- CD2 9)处理 HCC细胞 ,观察 Anti- CD2 9对细胞与 IV型胶原裱衬表面的黏附力的影响。采用双微吸管实验法进行 HCC细胞趋化实验 ,在两侧微管内加入相同浓度 (6 0 0μg/ ml )的 IV型胶原 ,并引导微管尖端与同一细胞紧密接触 ,动态观察细胞两侧伪足形成过程 ;在一侧微吸管内加入 2 0μg/ ml Anti- CD2 9,考察 beta1整合素亚单位阻断对 HCC细胞伪足形成的影响。利用流式细胞仪对 HCC细胞表面整合素 beta1亚单位的表达进行分析。结果表明 ,HCC细胞与 5μg/ ml IV型胶原裱衬表面之间的黏附力为 932± 134(× 10 - 1 0 N ,n=6 0 ) ,加入 5μg/ ml Anti- CD2 9黏附力减小到 4 4 9± 119(× 10 - 1 0 N,n=6 0 ) ;加入 10 μg/ ml Anti- CD2 9时黏附力减小到 2 2 0± 78(× 10 - 1 0 N,n=5 5 )。双微吸管趋化实验表明 :两侧微吸管加入相同浓度 IV型胶原 ,细胞向两侧微吸管均有伪足形成 ;在此基础上 ,微吸管加入 Anti- CD2 9的一侧 ,HCC细胞伪足生长曲线呈现明显的抑制 ,而未加入 Anti- CD2 9的微吸管一侧与加入的对侧相比 ,该侧细胞的伪足明显增     

Ⅳ型胶原和Laminin在人体慢性肝病和肝癌中的病理学研究

目的：观察基膜中Ⅳ型胶原（ＣｏⅣ）及Ｌａｍｉｎｉｎ（ＬＭ）在人体慢性肝病和肝细胞癌（ＨＣＣ）的病理形态学改变,并探讨其临床病理意义。方法：对１１例慢性肝炎、２６例肝硬化和３０例人体ＨＣＣ组织进行了ＣｏⅣ和ＬＭ的免疫组化检测,系统地观察基膜成分在正常、慢性肝病和肝癌中的形态和数量的改变,行半定量计数,对其结果进行临床病理分析。结果：正常肝窦无ＬＭ的表达,ＣｏⅣ沿窦壁呈断续弱阳性表达,慢性肝病时可有少     

核心蛋白聚糖对鼠肌腱细胞Ⅰ型胶原及其mRNA的影响

目的：探讨核心蛋白聚糖（Decorin，DCN）对大鼠肌腱细胞Ⅰ型胶原的影响，为进一步临床应用提供理论依据。方法：50μg/μL的DCN作用大鼠肌腱细胞12h后，采用免疫细胞化学，免疫荧光方法观察DCN对大鼠肌腱细胞Ⅰ型胶原蛋白的影响，采用半定量反转录聚合酶链反应（RT-PCR）方法检测DCN对大鼠肌腱细胞Ⅰ型胶原mRNA表达的影响。结果：DCN在早期能够明显减少肌腱细胞的Ⅰ型胶原蛋白的合成，免疫荧光定量分析实验组和对照组相差显著；DCN早期还能够显著下调肌腱细胞Ⅰ型胶原蛋白mRNA水平的表达。结论：DCN在早期对大鼠肌腱细胞Ⅰ型胶原的形成起着抑制作用。     

肺癌细胞与胞外基质选择裱衬表面粘附力学的研究

肿瘤细胞与细胞外基质的粘附特性与肿瘤的侵袭转移密切相关,作者力图揭示人肺癌细胞相应的生物力学和生物流变学特征,采用微管吸吮技术定量测定体外培养的低转移人肺腺癌（PAa_)细胞和高转移人肺巨细胞癌（PG）细胞与细胞外基质重要组份胶原蛋白IV和层粘连蛋白（LN）的粘附力学特性。结果表明：与胶原蛋白IV裱衬在的粘附力,在PA,aPG细胞均较裱衬前增加,但在较低胶原浓度（1．00ug/ml,2.00ug/ml)时PAa细胞的粘附力增加幅度大于PG细胞,与LN和固定浓度（2．00ug/ml)胶原蛋白IV的复合裱衬面的粘附力为PAa细胞大于PG细胞,在较低LN浓度,PAa(0.625ug/ml),PG(0.625ug/ml,1.25ug/ml)细胞粘附力相对降低,尤以PG细胞中的降低程度和幅度为大,因此,癌细胞与细胞外基质的粘附和去粘附行为主要通过膜受体介导,从而影响癌细胞生物学特性及侵润转移能力。     

烧伤大鼠粒细胞与内皮细胞粘附力学特性的研究

本文以大鼠作为烧伤的实验动物模型，以微管吸吮技术研究了烧伤情况下粒细胸（ＰＭＮ）与肺毛细血管内皮细胞（ＰＣＥＣ）的粘附力学特性．作者首先测定了ＰＭＮ与ＰＣＥＣ间的粘附力Ｆａ，进而在实验条件下定义了相对粘附应力Ｓ１．实验结果表明：在烧伤后２４小时内，粒细胞与肺毛细血管内皮细胞间的粘附力及相对粘附应力均显著升高，且在烧伤后１小时达到最大值。结合烧伤早期的病理变化，作者对实验结果进行了一定的分析和讨论．     

大鼠肺微血管内皮细胞培养及其粘弹性研究

为了建立肺微血管内皮细胞培养方法 ,研究肺微血管内皮细胞粘弹性。我们取大鼠肺周边组织 (宽度不应大于 1.5 mm) ,将组织剪成 1.5 mm× 1mm× 1mm的组织块 ,贴入无菌的 2 5 cm3培养瓶 ,每瓶 10～ 15块 ,同时加入含 2 0胎牛血清、肝素 90 U/ml、L-谷氨酰胺 4mmol、青霉素 10 0 U/ml和链霉素 10 0 μg/ml的 DMEM培养基 3ml,放入 37℃二氧化碳培养箱中静置培养 ;8h后翻转培养瓶 ,6 0 h后取出肺组织块 ,接着继续培养 2～ 4d后进行传代。最后消化分离肺微血管内皮细胞 ,用微管吸吮系统研究肺微血管内皮细胞粘弹性。结果显示 :肺微血管内皮细胞通过倒置相差显微镜观察 ,细胞呈鹅卵石镶嵌状排列 ,状如梭形或多角形 ,大小均匀 ,胞核清晰 ,呈卵圆形 ,胞浆丰富 ; 因子相关抗原免疫荧光染色呈阳性 ;肺微血管内皮细胞弹性模量 K1 =49.3± 9.2 Pa、K2 =73.2±2 4.8Pa、粘性系数 μ=19.2± 7.2 Pa.s。这些结果表明用组织块法培养肺微血管内皮细胞是可行的 ,肺微血管内皮细胞表现出较大的刚性     

Keratinocyte Differentiation and Proliferation are Regulated by Adhesion to the Three-Dimensional Meshwork Structure of Type IV Collagen

We examined the behavior of human foreskin keratinocytes (HFKs) on reconstituted type IV collagen gel. HFKs survived for several days and the upper layer cells expressed a differentiation marker, involucrin. Apoptosis was induced after involucrin expression while cell proliferation was suppressed. On molecular type IV collagen, integrins shifted from α2β1 to α3β1 during HFK culture. On type IV collagen gel, HFKs initially expressed integrin α2β1, and later expressed integrin α3β1 in the presence of α2β1 did not disappear. Using synthetic peptides, we examined integrin α2-mediated adhesion to type IV collagen gel. Addition of synthetic peptide dose-dependently inhibited cell adhesion both on type IV collagen gel and on molecular type IV collagen. On type IV collagen gel, weaker phosphorylation of focal adhesion kinase, paxillin, and Akt was observed compared with the molecular forms. Based on these observations, we think type IV collagen gel is a novel culture substrate that mimics the physiological environment for HFKs.     

Zonal Uniformity in Mechanical Properties of the Chondrocyte Pericellular Matrix: Micropipette Aspiration of Canine Chondrons Isolated by Cartilage Homogenization

The pericellular matrix (PCM) is a region of tissue that surrounds chondrocytes in articular cartilage and together with the enclosed cells is termed the chondron. Previous studies suggest that the mechanical properties of the PCM, relative to those of the chondrocyte and the extracellular matrix (ECM), may significantly influence the stress–strain, physicochemical, and fluid-flow environments of the cell. The aim of this study was to measure the biomechanical properties of the PCM of mechanically isolated chondrons and to test the hypothesis that the Young's modulus of the PCM varies with zone of origin in articular cartilage (surface vs. middle/deep). Chondrons were extracted from articular cartilage of the canine knee using mechanical homogenization, and the elastic properties of the PCM were determined using micropipette aspiration in combination with theoretical models of the chondron as an elastic incompressible half-space, an elastic compressible bilayer, or an elastic compressible shell. The Young's modulus of the PCM was significantly higher than that reported for isolated chondrocytes but over an order of magnitude lower than that of the cartilage ECM. No significant differences were observed in the Young's modulus of the PCM between surface zone (24.0 ± 8.9 kPa) and middle/deep zone cartilage (23.2 ± 7.1 kPa). In combination with previous theoretical biomechanical models of the chondron, these findings suggest that the PCM significantly influences the mechanical environment of the chondrocyte in articular cartilage and therefore may play a role in modulating cellular responses to micromechanical factors.     

125I粒子对食管癌Eca-109细胞凋亡及细胞周期的影响

研究,125I粒子诱导体外培养的人食管癌Eca-109细胞凋亡及对其细胞周期的影响.设置A组:对照(不加粒子),B组:低剂量(7.4×106Bq 125I粒子1枚),C组:中剂量(14.8×106Bq 125I粒子1枚),D组:高剂量(29.6×106Bq 125I粒子1枚),细胞培养1周后收集细胞,流式细胞术检测细胞凋亡及细胞周期.4组凋亡细胞阳性指数(AI)分别为0.17%、1.17%、4.35%、4.72%,B、C和D组与A组之间均有非常显著性差异(P<0.01);C组与B组,D组与B组比较,均有非常显著性差异(P<0.01),D组与C组比较无显著性差异(P>0.05).随粒子剂量增加,G2/M期细胞所占比例增高,各组之间有非常显著性差异(P<0.01).125I粒子可以诱导体外培养的人食管癌Eca-109细胞凋亡,其诱导凋亡的能力可能是通过把细胞阻滞在G2/M期实现.     

Cell Death and Proliferation and Its Relation to Collagen Degradation in Uterine Involution of Rat

Collagen concentration, procollagenase localization, and their association with cell proliferation and apoptosis during postpartum involution, were investigated biochemically and histochemically in postpartum day 1, 3, 5, and 7 rat uterine tissues. In control animals, uterine wet weight, soluble protein, and collagen decreased rapidly during days 1 to 3 postpartum, and the DNA concentration in the uterine horn rapidly decreased, as noted by others. Simultaneously, both apoptosis and cell proliferation were observed in these tissues. These processes were highest in smooth muscle cells on day 3 postpartum. Procollagenase was found in the cell cytoplasm through days 1 to 3 postpartum, was highest on the third day postpartum, and appeared to gradually diminish by day 5 postpartum. Disorganization of collagen fibers was observed, under polarized microscopy by a strong birefringence of collagen fibers of the circular smooth muscle cell layers. However, this disorganization of the uterine collagen diminished progressively from day 3 to day 7. Treatment with estradiol or a combination of estradiol and progesterone suppressed cellular turnover and attenuated the changes in DNA, total amino acids, and collagen on day 3 postpartum. In this study, cellular turnover and biochemical and morphological changes appeared to be closely associated. Gonadal steroid hormones appear to influence these changes and retard uterine involution. This study suggests that a dynamic turnover of the cellular population takes place during uterine involution. It is possible that other factors, in addition to steroid hormones, contribute to uterine involution. It is to be postulated that these factors either are themselves decreased or, alternatively, may increase the inhibition of other unknown factors by an indirect mechanism.     

131I治疗Graves病的转归与三种细胞因子的关系

探讨Graves病131I治疗后的转归与细胞因子IL-2、IL-10、TNF-α之间的关系.158例Graves病首次接受131I治疗的患者在治疗前和治疗后5个月分别抽取静脉血,检测IL-2、IL-10、TNF-α和T3、T4、FT3、FT4和s-TSH.根据治疗后5个月T3、T4、FT3、FT4和s-TSH含量分为甲亢未控制组、甲亢控制组和早发性甲减组.结果显示,IL-2、IL-10各组患者治疗前、后均明显高于对照组(P＜0.01),TNF-α早发性甲减组治疗前、后均高于对照组(P＜0.05).IL-2治疗前后组间比较,甲亢未控制组最高,甲亢控制组次之,早发性甲减组最低,三组治疗后均高于治疗前(P＜0.05);IL-10治疗后甲亢未控制组明显高于其余两组(P＜0.01);THF-α治疗前后组间比较,早发性甲减组高于其余两组(P＜0.01).IL-2/IL-10比值治疗前后组间比较,甲亢未控制组最高、甲亢控制组其次,早发性甲减组最低.作者认为,Graves病患者机体免疫状态处于一个较高的水平,以Th1细胞因子优势表达为主.IL-2/IL 10比值越高,越难控制,IL-2/IL-10比值越低越易产生甲减.Graves病患者治疗前、后TNF-α含量高的患者容易发生早发性甲减.     

mB7-1转染大鼠卵巢癌细胞诱导细胞免疫应答及体内致瘤性研究

为探讨转染mB7 1基因大鼠卵巢癌细胞系后体外诱导细胞免疫应答及其致瘤性 ,我们以逆转录病毒为载体将mB7 1基因转染入大鼠低分化卵巢上皮癌细胞株NuTu 19中 ,流式细胞仪检测mB7 1的表达。用同源淋巴细胞肿瘤细胞混合培养实验 (MTLCs)测定淋巴细胞增殖指数 ,MTT法检测细胞毒淋巴细胞的增殖及其杀瘤活性。将NuTu 19/Neu、NuTu 19/mB7 1细胞分别接种于同源大鼠Fischer344皮下 ,观察mB7 1修饰的肿瘤细胞在动物体内的致瘤性。结果显示 ,mB7 1基因成功地转染入大鼠低分化卵巢上皮癌细胞株NuTu 19,流式细胞仪测定mB7 1基因呈阳性表达。NuTu 19/mB7 1体外刺激脾淋巴细胞增殖能力明显高于对照组细胞 (P <0 0 5 )。NuTu 19/mB7 1体外诱导的CTL较对照组对NuTu 19细胞的杀伤率显著增高 (P<0 0 1)。两种细胞体外增殖能力无明显改变 ,NuTu 19/mB7 1组在Fischer344大鼠体内成瘤时间明显晚于对照组 (P <0 0 5 )。由此可见 ,mB7 1基因修饰的鼠卵巢癌细胞可体外诱导细胞免疫应答 ,接种动物后 ,实验瘤苗的致瘤性下降