五氧化二钒对大鼠胚胎肢芽细胞增殖和分化的影响

本研究采用胚胎肢芽细胞微团培养法，通过体外直接染毒及经体内染毒后在体外观察两种方式，观察了Ｖ２Ｏ５对大鼠胚胎肢芽细胞增殖和分化的影响。结果显示，Ｖ２Ｏ５对体外肢芽细胞的增殖和分化均有明显抑制作用，并有较明显的剂量—反应关系，其ＩＣ５０分别为１３．６４和４．７７μｍｏｌ／Ｌ。体内／体外结合试验结果，Ｖ２Ｏ５对细胞增殖无明显影响，但对细胞分化有明显抑制作用。结果表明，Ｖ２Ｏ５对大鼠胚胎肢芽细胞有特异分化抑制作用。     

利用全胚胎和微团培养模型评价硝酸铈的发育毒性

目的：利用大鼠植入后全胚胎培养模型和微团培养模型模拟胚胎发育早期和中、晚期过程,评价硝酸铈的发育毒性,以期为进一步开展体内实验研究及合理制定人群稀土接触的健康指导值提供科学依据。方法：取孕9.5 d的SD大鼠胚胎,分别在含0.00、0.50、0.75、1.00 mmol/L硝酸铈的大鼠即刻离心血清中37℃旋转培养,48 h后根据Brown's评分法对胚胎的生长发育及形态功能进行评分,结合BALB/c 3T3细胞毒性结果,利用欧洲替代方法验证中心（ECVAM）全胚胎预测模型评价硝酸铈胚胎早期发育毒性。分离孕13 d的SD大鼠胚胎肢芽原代细胞进行微团培养,分别用0.03、0.06、0.13、0.25、0.50、1.00、2.00和3.00 mmol/L硝酸铈进行染毒,培养5 d后利用中性红活细胞摄取染色法测定50%细胞增殖受抑制时的浓度（IC50）,利用阿利新蓝染色法测定50%细胞分化受抑制时的浓度（ID50）,根据ECVAM微团培养预测模型评价硝酸铈胚胎中、晚期发育毒性。结果：全胚胎培养模型中硝酸铈对胚胎生长的无可见有害作用水平（NOAEL）为0.50 mmol/L,0.75 mmol/L以上浓度硝酸铈可显著降低胚胎卵黄囊直径和顶臀长（P < 0.05）,抑制胚胎生长,并可致胚胎体节数目减少（P < 0.05）,胚胎发育畸形。微团培养模型中硝酸铈对肢芽细胞增殖活性的NOAEL为0.25 mmol/L,对肢芽细胞分化为软骨细胞的NOAEL为0.12 mmol/L,其IC₅₀和ID₅₀分别为1.23和0.76 mmol/L。结论：经全胚胎培养预测模型和微团培养模型评价硝酸铈为弱发育毒性化学物。     

利用全胚胎和微团培养模型评价硝酸镧的发育毒性

目的： 利用大鼠植入后全胚胎培养模型和微团培养模型评价硝酸镧潜在的胚胎早期和中、晚期发育毒性。方法： 将大鼠胚胎随机分为5组后培养于含不同剂量硝酸镧（0、0.12、0.23、0.46及1 mmol/L）的即刻离心血清中，48 h后测量胚胎卵黄囊直径（YSD）、顶臀长（CRL）和头长（HL），计数体节，并根据Brown's评分法对胚胎进行总形态学评分（TMS）。同时，对BALB/c 3T3细胞进行细胞毒性测试。根据欧洲替代方法验证中心（ECVAM）的预测模型对硝酸镧的胚胎早期发育毒性进行评价。分离大鼠胚胎肢芽细胞，制成单细胞悬液，进行微团培养。分别用0.03、0.06、0.13、0.25、0.5、1、2及3 mmol/L硝酸镧进行染毒，利用中性红活细胞摄取染色法测定50%细胞增殖受抑制时的浓度（IC₅₀）以反映细胞增殖情况，利用阿利新蓝染色法测定50%细胞分化受抑制时的浓度（ID₅₀）以反映细胞分化情况。根据ECVAM预测模型对硝酸镧胚胎中晚期发育毒性进行评价。结果： 硝酸镧对胚胎生长的无可见有害作用水平（NOAEL）为0.12 mmol/L，0.46 mmol/L硝酸镧可明显降低胚胎YSD、CRL和HL（P < 0.05），对胚胎生长有明显的抑制效应。0.46 mmol/L硝酸镧可诱导胚胎产生发育畸形，减少胚胎体节形成数目（P < 0.05）。硝酸镧对肢芽细胞增殖活性的NOAEL为1 mmol/L，对肢芽细胞分化为软骨细胞的NOAEL为0.25 mmol/L，其IC₅₀和ID₅₀分别为1.57 mmol/L（510.25 μg/mL）和0.99 mmol/L（321.75 μg/mL）。结论： 经全胚胎培养预测模型评价硝酸镧为弱胚胎发育毒性化学物；经微团培养模型评价硝酸镧为无胚胎发育毒性化学物。     

采用微团培养模型探讨染料木黄酮的发育毒性

目的：利用大鼠胚胎肢芽和中脑细胞微团培养模型研究染料木黄酮（genistein,GEN）的发育毒性,并探讨其发育毒性机制。方法：实验分为不同浓度的GEN（0、0.94、1.875、3.75、7.5、15.0μg/ml）染毒组,受试物分别作用于培养大鼠胚胎肢芽细胞和中脑细胞微团,采用中性红摄取法检测GEN对细胞增殖的影响,采用阿利新蓝染色方法检测GEN对肢芽细胞分化的影响,采用图像处理分析方法检测GEN对中脑细胞分化的影响。计算细胞50%增殖抑制浓度（IC50-P）和50%分化抑制浓度（IC50-D）。用不同浓度的雌激素受体拮抗剂ICI182780（0.1,0.5,1μmol/L）分别预处理细胞后再加入GEN（7.5μg/ml）,观察雌激素受体途径在GEN诱导的发育毒性中的作用。结果：GEN对肢芽细胞的IC5-0P和IC5-0D分别为5.4μg/ml和4.9μg/ml,GEN对中脑细胞的IC5-0P为6.2μg/ml,IC5-0D分别为7.1μg/ml（集落分化个数）或5.3μg/ml（集落分化面积）。IC5-0P/IC50-D的比值均接近1。在GEN7.5μg/ml染毒组,用ICI182780预处理细胞,不能改变GEN诱导的发育毒性作用。结论：根据Flint＇s和欧洲替代方法验证中心ECVAM致畸物判别标准,并结合人体实际可能接触水平,认为GEN为强致畸物,并且对人类存在可能的风险。其致畸作用可能主要通过细胞毒性发挥作用,不依赖于雌激素受体途径。     

利用全胚胎和微团培养模型评价硝酸镧的发育毒性

目的： 利用大鼠植入后全胚胎培养模型和微团培养模型评价硝酸镧潜在的胚胎早期和中、晚期发育毒性。方法： 将大鼠胚胎随机分为5组后培养于含不同剂量硝酸镧（0、0.12、0.23、0.46及1 mmol/L）的即刻离心血清中,48 h后测量胚胎卵黄囊直径（YSD）、顶臀长（CRL）和头长（HL）,计数体节,并根据Brown's评分法对胚胎进行总形态学评分（TMS）。同时,对BALB/c 3T3细胞进行细胞毒性测试。根据欧洲替代方法验证中心（ECVAM）的预测模型对硝酸镧的胚胎早期发育毒性进行评价。分离大鼠胚胎肢芽细胞,制成单细胞悬液,进行微团培养。分别用0.03、0.06、0.13、0.25、0.5、1、2及3 mmol/L硝酸镧进行染毒,利用中性红活细胞摄取染色法测定50%细胞增殖受抑制时的浓度（IC₅₀）以反映细胞增殖情况,利用阿利新蓝染色法测定50%细胞分化受抑制时的浓度（ID₅₀）以反映细胞分化情况。根据ECVAM预测模型对硝酸镧胚胎中晚期发育毒性进行评价。结果： 硝酸镧对胚胎生长的无可见有害作用水平（NOAEL）为0.12 mmol/L,0.46 mmol/L硝酸镧可明显降低胚胎YSD、CRL和HL（P < 0.05）,对胚胎生长有明显的抑制效应。0.46 mmol/L硝酸镧可诱导胚胎产生发育畸形,减少胚胎体节形成数目（P < 0.05）。硝酸镧对肢芽细胞增殖活性的NOAEL为1 mmol/L,对肢芽细胞分化为软骨细胞的NOAEL为0.25 mmol/L,其IC₅₀和ID₅₀分别为1.57 mmol/L（510.25 μg/mL）和0.99 mmol/L（321.75 μg/mL）。结论： 经全胚胎培养预测模型评价硝酸镧为弱胚胎发育毒性化学物;经微团培养模型评价硝酸镧为无胚胎发育毒性化学物。     

羟基脲对大鼠胚胎中脑细胞增殖和分化的影响

背景与目的:为探讨羟基脲(HU)对胚胎发育的作用,采用胚胎中脑细胞微团培养研究HU对细胞增殖和分化的影响.材料与方法:从13 d龄的鼠胚胎中分离中脑细胞,加入不同浓度的HU体外连续培养5 d,观察细胞分化和细胞增殖情况.结果:对照组原单个分散的细胞形成明显的细胞集落,集落间有丰富的神经纤维,并连结成网状,而不同HU剂量组细胞集落数目减少,集落间的神经纤维束减少,并呈现出剂量-效应关系,显示HU可明显抑制中脑细胞的增殖和分化,阻止神经形成和集落形成过程,HU半数细胞增殖抑制浓度(IcV50)为0.078 mmol/L(5.9 μg/ml),半数细胞分化抑制浓度(IcD50)为0.018 mmol/L(1.37μg/ml),其IcV50与IcD50比值为4.33.结论:HU是一种非特异性细胞增殖和分化抑制剂,具有体外细胞致畸作用.     

体外培养技术及其在头颈部鳞状细胞癌研究中的应用

 头颈肿瘤占全身肿瘤的5 % ,其中90 %是恶性肿瘤。头颈肿瘤具有独特的特征,如癌前病变易于识别;上气消化道(upperaerodigestivetract)肿瘤常位于体表,较易监测;流行病学资料显示能从肿瘤发生的部位和分期来预测局部和远处转移情况;原发肿瘤易复发并有继发其他部位肿瘤的危险性;另外,头颈部肿瘤是用化学药物预防肿瘤复发的理想模型。头颈肿瘤的这些特点决定了其在人体肿瘤研究中所具有的重要地位。当前,尽管头颈部鳞状细胞癌研究已取得重大进展,但随着研究的广泛和深入,出现愈来愈多待解决的问题。较多证据表明,体外培养技术的应用可望为头颈部鳞状细胞癌发生发展机制的阐明和防治带来新契机。本文将介绍这一领域的进展。1　体外培养技术概述体外培养(InVitroculture)是指从体内取出组织或器官摹拟体内生理环境,在无菌、适当温度和一定营养条件下,使之生存和生长并维持其结构和功能的方法。包括细胞培养(CellCulture)、组织培养(TissueCulture)和器官培养(OrganCulture)。组织培养和细胞培养并无严格区别。培养过程中,不论培养物是组织还是细胞都要生活在人工环境中,长期的培养易导...     

三维培养技术的研究进展及其在骨肿瘤研究中的应用

三维培养技术是相对传统二维平面培养的一种培养技术,通过提供相应的细胞基质,构建细胞、组织或器官的模拟三维微环境,从而实现细胞与组织基质细胞相互作用的培养模式.传统的二维平面培养最主要的缺陷是缺少细胞基质,不能形成有效的空间结构,无法模拟正常物质代谢运输及内分泌作用.三维培养技术在准确再现肿瘤立体结构的同时,将为肿瘤研究者进行肿瘤靶向治疗研究提供更精确的研究对象,促进研究者对肿瘤细胞相关的基质细胞信号通路及基因表达模型深入研究.     

肿瘤细胞培养过程中的吸收光谱研究

目的　寻找到一种标志癌症转移情况的有效方法 ,为转移学的研究提供证据。方法　用吸收光谱的方法分别采集了相同培养天数的CNE细胞和HeLa细胞。结果　发现两者的吸光度在 2 0 0～ 3 0 0毫微米处均有特征性的谱线 ,另外发现它们的吸光度有差异 ,且它们各自的吸光度与细胞的培养天数无正比关系。结论　不同的肿瘤细胞谱线模型不同 ,预示不同癌症的转移进程存在差异。     

小鼠胎盘周细胞的原代培养及其在胎盘血管毒物筛查中的初步应用

目的:建立小鼠胎盘周细胞的分离、培养方法并应用于胎盘血管毒物筛查研究。方法:将C57小鼠雌雄合笼受孕后,在小鼠妊娠的第18.5天,取胎盘,通过机械分离和组织消化法分离、培养小鼠胎盘周细胞;采用免疫荧光法鉴定细胞来源与纯度;采用MTS比色法检测胎盘周细胞的生长曲线。将胎盘周细胞分别暴露于0、6.25、12.50、25.00、50.00和100.00 μmol/L的醋酸铅24 h后,用MTS法检测染毒后的细胞活力。结果:利用机械分离和组织消化法分离、培养后,小鼠胎盘周细胞可贴壁生长,呈多边形,表达特异性标志α-平滑肌肌动蛋白(α-SMA)。在暴露于6.25~100.00 μmol/L醋酸铅24 h后,随着染毒剂量的增加,细胞的存活率逐渐下降,且各个剂量组之间的差异具有统计学意义(P<0.05)。结论:采用机械分离和组织块消化法可以简单、快捷地获得高纯度的小鼠胎盘周细胞,并可以应用于铅等胎盘血管毒物的检测。     

五氧化二砷对大鼠胚胎肢芽细胞增殖和分化的影响

The micromass culture was used to determine the effects of vanadium pentoxide(v2O5)on the proliferation and differentiation of limb bud cells of rat．In the in vitro test,the results showed that V2O had obvious inhibiting effects on both proliferation and difierentiation of limb bud cells with a dose-dependent response,its proliferating and differentiating IC50 being 13．64 and 4．77 Vmol／L,respectively． In the in vivo/in vitro test,the results showed that V2O5 had no obvious effect on cell proliferation but had obvious inhibiting effect on cell differentiation．These results indicated that V205 might have a specific inhihifing effec t on the differentiation oflim b bud cells．     

A New Rat Colon Cancer Cell Line Metastasizes Spontaneously: Biologic Characteristics and Chemotherapeutic Response

A new cell line (RCN-9) was established in culture from a transplantable rat colon adenocarcinoma, which was induced in the colon of a male Fischer F344 rat by subcutaneous administration of 1,2-dimethylhydrazine. When RCN-9 cells were injected subcutaneously or into the cecal subserosa of syngeneic rats, carcinomas with progressive growth were obtained and the development of lung (63.6%) and liver (40.0%) metastases, respectively, ensued. Antitumor effects of 5-fluorouracil (5-FU), adriamycin (ADM) and mitomycin C (MMC) against RCN-9 were examined in vivo and in vitro. 5-FU and ADM had antitumor effects both in vivo and in vitro; MMC had antitumor effects in vitro. These results show that the RCN-9 cell line can be used both as a model to study mechanisms of metastasis from colon carcinoma and as a model in chemotherapeutic studies of metastatic disease from colon carcinoma.     

Monoclonal Antibody against Rat Renal Cell Tumor-associated Antigen as a New Tool for the Analysis of Renal Tumorigenesis

A monoclonal antibody, NR-2 MAb, against one of the rat renal cell tumor-associated antigens was developed. NR-2 MAb belonged to IgM class and recognized a polypeptide of 81,000 daltons designated as NR-2 antigen, which is of non-glycoprotein nature with a pI of 4.6. NR-2 MAb was employed to probe the histogenesis of renal cell tumors in rats treated with N-ethyl-N-hydroxyethylnitrosamine followed by trisodium nitrilotriacetate monohydrate. Immunohistochemical analysis indicated that NR-2 antigen was expressed in simple hyperplasia, adenomatous hyperplasia and renal cell tumors. Both clear cells and basophilic cells of the simple hyperplasia showed equally strong positive reactions with NR-2 MAb, whereas the vacnolated epithelium was negative. Furthermore, the proximal tubules in nontumorous areas also expressed NR-2 antigen, suggesting that the hyperplastic lesions which eventually lead to renal cell tumors may derive from cpithelia of proximal tubules and not directly from vacuolated epithelium. Such NR-2 antigen-positive epithelia of proximal tubules seem to be initiated cells. NR-2 MAb also cross-reacted with preneoplastic liver lesions.     

人上颌窦癌体外细胞培养初步报告

我们将11例人上颌窦低分化鳞状细胞癌进行体外细胞培养,除1例未生长外,其余10例均见生长。培养人癌细胞存活时间。最长者达18天零3小时,最短苦为8天。其中1例在作直接培养的同时,取部分组织先经裸小鼠体内移植生长后,再作体外细胞培养,培养细胞最长存活时间达27天,最短存活时间为10天。实验表明,体外培养人上颌窦癌细胞在一定时间内生长良好,可望建立单纯稳定的细胞系,并为进一步探讨上颌窦癌的生物学特性及进行药物筛选研究提供有利条件。     

骨水泥在肱骨近端骨巨细胞瘤术后骨缺损修复中的应用

目的 探讨骨水泥塑型骨段修复重建肱骨近端骨巨细胞瘤术后骨缺损的临床应用价值.方法 对5例原发骨巨细胞瘤患者的治疗情况进行回顾性分析.结果 全部患者随访24～84个月,平均44个月,均无复发和转移;根据Ermeking的骨骼肌肉肿瘤术后功能评分,本组5例患者术后患侧上肢功能评分20～28分,平均26分,其中评分在23分以...     

Coexpression of Multiple Sertoli Cell and Leydig Cell Marker Genes in the Spontaneous Testicular Tumor of F344 Rat: Evidence for Phenotypical Bifurcation of the Interstitial Cell Tumor

The development of testicular tumor has been frequently observed in some laboratory rat strains. In the present study, we have further characterized the testicular tumor that spontaneously develops in the F344 rat (F344/Jcl). Tumor cells first appeared in the interstitium and developed into multifocal nodular lesions. In the later stage, the whole testes were occupied by tumor cells that consisted of three different types of cells in morphological appearance: large clear type, small eosinophilic type and intermediate type. To determine the character of these cells, we examined the expression of marker genes for Sertoli cells (e.g., transferrin) and Leydig cells (e.g., 3β–hydroxysteroid dehydrogenase 1 (3β–HSD 1)). Transferrin and 3β–HSD 1 mRNAs were found in all 8 tumor samples analyzed hy northern blotting. By in situ hybridization, we observed a substantial amount of 3β–HSD 1 mRNA and little or no transferrin mRNA in the large clear cells. In contrast, the small eosinophilic cells showed little or no 3β–HSD 1 mRNA and a large amount of transferrin mRNA, suggesting that the tumor was a mixture of at least two types of cells. Other Sertoli cell marker genes, such as cyclic protein 2 and sulfated glycoprotein 2, were expressed in all 8 tumors analyzed, and testin and steel factor (SLF), the c–kit receptor ligand, were also expressed in some of the tumors (testin, 75%; SLF, 25%), while other Leydig cell markers, LH receptor and c– kit , were expressed in 87% and 80% of the tumors, respectively. These results indicate that the spontaneous testicular tumor of F344 rat is of interstitium origin, showing phenotypical bifurcation possibly via transdifferentiation.     

Induction of DNA Replication and Cell Growth in Rat Liver by Obstructive Jaundice

Obstructive jaundice, produced by ligating the common bile duct, induced a transient DNA replication followed by cell proliferation in rat liver. At 48 h after the operation, DNA polymerase α activity started to increase and reached its maximum level (more than twice the control) at day 4. At day 7, the enzyme level had decreased to the control level. Pulse-labeling experiment using radioactive thymidine showed that the rate of DNA synthesis increased approximately 2.5-fold in the same pattern as that of DNA polymerase α The mitotic index in hepatocytes also increased 10-fold at day 4 and then decreased. The proliferation of liver cells induced by obstructive jaundice mimics the regeneration of partially hepatectomized liver, although the response was slightly delayed and the proliferation was transient.