用抗溴脱氧尿苷单克隆抗体检测小鼠各种器官内S期细胞…

用溴脱氧尿苷（ＢｒｄＵ）腹腔注入正常小鼠（０．１５ｍｇ／ｇ体重），１ｈ后处后，取出多种器官的组织用Ｃａｒｎｏｙ液固定，石蜡包埋切片。用免疫组织化学技术，以抗－ＢｒｄＵ的单克隆抗体检测不同器官内被ＢｒｄＵ标记的Ｓ期细胞。结果证明，ＢｒｄＵ阳性细胞主要分布在细胞增殖活跃的组织，如舌与食管的基底层细胞、胃小凹处的颈粘液细胞，小、大肠的肠腺细胞和睾丸曲细精管内的精原细胞等。ＢｒｄＵ阳性细胞在淋巴器官内主要     

用BrdU标记技术观察大鼠脊髓上胸段灰质细胞的分化发育

目的 观察脊髓灰质细胞的分化发育,寻找脊髓移植时胎鼠脊髓取材的最佳时间。方法 用５－溴－２脱氧尿嘧啶（ＢｒｄＵ）标记ＤＮＡ,ＡＢＣ免疫细胞化学方法观察大鼠上胸段脊髓灰质细胞的增殖分化情况。结果 孕鼠于Ｅ１０ｄｅ腹腔注射ＢｒｄＵ,胎刀脊髓上胸段ＢｒｄＵ免疫反应神经元主要集中在脊髓后角,脊髓侧角的ＢｒｄＵ免疫反应神经元较少,前角仅有少量的ＢｒｄＵ免疫反应神经元;孕鼠于Ｅ１１ｄ腹腔注射ＢｒｄＵ,胎鼠脊髓     

人胎儿胸腺T细胞亚群抗原的表达及其定位—免疫组织化学研究

用免疫组织化学法和多种识别Ｔ细胞亚群的抗体，观察了１５例胎儿胸腺内Ｔ细胞亚群的抗原表达及其分布。结果表明，胸腺皮、髓质Ｔ细胞对Ｌｅｕ４抗体均呈阳性，但髓质部较强。Ｌｅｕ３ａ和Ｌｅｕ２ａ阳性细胞分布于胸腺皮持、髓质，但髓质部阳性数目少于皮质。ＡＩＧ３和ＮＫ阳性细胞均分布于髓质，皮质几乎为阴性，ＩＬ－２Ｒ细胞分布于被膜下、皮质和髓质。ＢｒｄＵ标记的细胞主要存在于皮质，特别是外皮质区。此外胸腺内还有γδ     

CXCL13被证实为滤泡树突细胞肉瘤的一种新的标记物

内环境稳定性趋化因子CXCL13主要产生于B细胞聚集的淋巴滤泡，通过结合表达选择性受体CXCR5的B淋巴细胞的方式在淋巴器官的发育过程中发挥关键作用。滤泡树突细胞（FDCs）已被证实是淋巴样器官内产生该趋化因子的主要细胞群体。最近，全基因组检测方法提示淋巴滤泡内CD4T辅助细胞[T（h）F]是生发中心内另一种产生CXCL13的细胞群体，并且T（h）F的肿瘤性类似物（血管免疫母T细胞淋巴瘤中CD4＋的肿瘤性T细胞）仍保留产生这种趋化因子的能力。     

抗BrdU单克隆抗体的制备及初步鉴定

溴脱氧尿嘧啶核苷（Ｂｒｏｍｏｄｅｏｘｙｕｒｉｄｉｎｅ　ＢｒｄＵ）是ＤＮＡ前体胸腺嘧啶核苷的类似物。通过检测ＢｒｄＵ掺入Ｓ期细胞ＤＮＡ的基因片段,可避免用同位素标记所产生的放射性污染等副作用。本文在过碘酸氧化法偶联抗原的基础上,已制备出具有均一分泌活性...     

神经营养素在小鼠脾的定位研究

为了解神经营养素与免疫系统的关系，用免疫组织化学方法对神经营养素包括神经生长因子（ＮＧＦ）、脑源性神经营养因子（ＢＤＮＦ）、神经营养素３（ＮＴ－３）进行小鼠脾的定位研究。结果表明：３种神经营养素的免疫反应均存在于脾内，但分布特点各不相同。ＮＧＦ主要分布于白髓动脉周围淋巴鞘（ＰＡＬＳ）外层、边缘区（ＭＺ）和红髓（ＲＰ）的巨噬细胞样和淋巴细胞样细胞；ＢＤＮＦ除具有与ＮＧＦ相似的分布特点外，还见于脾淋巴小结生发中心的淋巴细胞样细胞；ＮＴ－３则存在于白、红髓的网状细胞样细胞。这一结果提示，脾的免疫和非免疫细胞均可能产生神经营养素，并提示不同类型的神经营养素对免疫系统有不同的作用。     

BrdU单克隆抗体的制备及鉴定

ＤＮＡ的５－溴，２－脱氧尿嘧啶核苷（ＢｒｄＵ）标记法近年来作为一种新的、简便、敏感特异的检测方法，在肿瘤生物学、遗传学、分子生物学特别是细胞增殖动力学等领域得到广泛应用，在一定程度推动了相关学科的发展。为了满足广泛开展ＢｒｄＵ标记ＤＮＡ检测对Ｂｒｄ...     

人胎儿期肠系膜淋巴结内T,B淋巴细胞分布的免疫组织化学研究

收集１８例第１２－４０周人胎儿肠系膜淋巴结，用免疫组织化学技术，观察了Ｔ、Ｂ淋巴细胞的分布。结果显示：ｓＩｇＭ和Ｋａｐｐａ轻链阳性Ｂ细胞主要分布在淋巴结的皮质部。Ｔ细胞分布较广泛，除融皮质区和皮质浅层外，还大量集中在髓质部。第２９周后，洒巴结、Ｂ细胞在皮质部已形成初级淋巴小结，但无生发中心和浆细胞。淋巴结内的毛细血管后微静脉随胎龄的增长而增多，其意义可能与介导淋巴细胞进入淋巴结内Ｔ、Ｂ细胞的分布区     

支气管扩张症神经内分泌和免疫活性细胞的变化

应用免疫组化方法对３５例支气管扩张症和１０例正常肺组织中肺内分泌细胞和免疫活性细胞进行观测，并对人的支气管相关淋巴组织（ＢＡＬＴ）进行形态学和免疫组化观察。结果显示：支气管扩张症中，支气管上皮降钙素和五羟色胺阳性细胞显著增多，支气管周围ＩｇＧ、ＩｇＡ和ＩｇＭ阳性细胞以及ＵＣＨＬ，阳性Ｔ细胞显著增多，ＢＡＬＴ明显增生，且ＢＡＬＴ增生的区域肺内分泌细胞和免疫活性细胞增多尤为显著，提示支气管扩张症发病中有神经内分泌和免疫系统参与。     

支气管扩张症神经内泌和免疫活性细胞的变化

应用免疫组化方法对３５例支气管扩张症和１０例正常肺组织中肺内分泌细胞和免疫活性细胞进行观察，并对人的支气管相关淋巴组织（ＢＡＬＴ）进行形态学和免疫组化观察。结果显示：支气管扩张症中，支气管上皮降钙素和五羟色胺阳性细胞显著增多，支气管周围ＩｇＧ，ＩｇＡ和ＩｇＭ阳性细胞以及ＵＣＨＬ１阳性Ｔ细胞显著增多，ＢＡＬＴ明显增生，且ＢＡＬＴ增生的区域肺内分泌细胞和免疫活性细胞增多尤为显著，提示支气管扩张症发病中     

Inhibition of biological effects of bromodeoxyuridine by deoxycytidine: Correlation with decreased incorporation of bromodeoxyuridine into DNA

The effects of 5-bromodeoxyuridine (BrdU) on pigmentation, contact inhibition, cell morphology, and tumorogenicity of Syrian hamster melanoma cells are inhibited in the presence of deoxycytidine (dC). The inhibition of these biological effects of BrdU by dC is correlated with a decrease in the incorporation of BrdU into nuclear DNA. The results suggest that the intracellular changes resulting from the addition of dC to cells in the presence of BrdU are comparable to those resulting from a decrease in the concentration of BrdU in the medium.     

The mechanism of the diurnal periodicity of mitosis

Summary Inversion of the alternation of light and darkness during the 24-hour period resulted in a complete inversion of the diurnal rhythm of mitotic activity in the epithelium of the skin, cornea, tongue and esophagus. After removal of the adrenal or thyroid glands the diurnal fluctuations of the number of mitoses were preserved, but their range in the morning and evening hours was somewhat reduced. Diurnal fluctuations of mitotic activity, absent in control animals, were noted in mice fed only at night. When mice were fed only in the morning the diurnal fluctuations were changed in the interstinal and tongue epithelium. The diurnal fluctuation of the RNA and DNA content noted in the cells of different organs corresponded to the mitotic rhythm. When the animals were illuminated at night the time of the maximum nucleic acid content changed from the morning to the evening.Presented by Active Member AMN SSSR V. N. Chernigovskii     

Detection of cellular receptors for Sendai virus in mouse tissue sections

We have established a method for detecting virus receptors in the tissue sections and have been able to clarify the distribution of cellular receptors for Sendai virus in various mouse organs, including trachea, esophagus, large intestine and cerebrum. The virus receptors were detected in ciliated cells of the trachea, as to be expected for a pneumotropic virus. The lamina propria and connective tissue of the mouse esophagus and large intestine, immediately under the epithelium, contained an abundance at virus receptors, but very few were detected in the epithelium of these organs. In mouse cerebrum, the cells lining the ventricles had a large number of virus receptors.     

Effects of thymidine analogs on Syrian hamster melanoma cells: Phenotypes arising from selection for analog resistance

In order to compare the biological effects of different thymidine (dT) analogs, two unusual cell lines (B-4 and HAB) previously isolated from a Syrian hamster melanoma line by selection with 5-bromodeoxyuridine (BrdU) were analyzed for their response to other analogs. B-4 cells require high concentrations of BrdU for optimal growth, and it was seen that the requirement for BrdU could be satisfied partially by 5-chlorodeoxyuridine (CldU) but not by the other dT analogs tested. HAB cells are able to grow with all the dT residues in nuclear DNA replaced by BrdU, and it was found that they could also grow with essentially all the dT residues in nuclear DNA replaced by CldU but not by other analogs. New cell lines resistant to 100 M concentrations of CldU, 5-iododeoxyuridine (IdU), and 5-hydroxymethyldeoxyuridine (HMdU) were isolated from the melanoma line and tested for cross-resistance to the other dT analogs. A high level of cross-resistance was observed only with BrdU and CldU. The ability of the cell lines resistant to BrdU, CldU, and IdU to incorporate these analogs into nuclear DNA also was determined. BrdU and CldU were incorporated efficiently by all of the lines tested, but the IdU-resistant cells seemed to preferentially exclude IdU.     

Relationship of DNA strand breakage produced by bromodeoxyuridine to topoisomerase II activity in Bloom-syndrome fibroblasts

Cells from patients with Bloom syndrome, a cancer-prone disorder with cutaneous photosensitivity and spontaneous chromosome breakage, exhibit an abnormally increased number of sister-chromatid exchanges following treatment with 5-bromodeoxyuridine (BrdU). This effect has been postulated to be mediated by abnormal topoisomerase II activity. We used alkaline elution to measure DNA single-strand breakage following prolonged exposure to BrdU. Five-day exposure to BrdU produced equal numbers of alkali-labile sites in normal and Bloom-syndrome fibroblasts. These breaks were not protein-associated but were produced by alkali. Treatment with topoisomerase II inhibitors induced similar frequencies of DNA single-strand breaks in normal and Bloom-syndrome fibroblasts. These findings imply that BrdU incorporation into cellular DNA induces alkali-labile DNA lesions that are independent of topoisomerase II activity in Bloom and normal cells.     

The assessment of cell proliferation during 9,10-dimethyl-1,2-benzanthracene-induced hamster tongue carcinogenesis by means of histone H3 mRNA in situ hybridization

The aim of this study was to investigate cell kinetics and ultrastructural changes during carcinogenesis using a hamster 9,10-dimethyl-1,2-benzanthracene (DMBA)-induced tongue cancer model. Five squamous cell carcinomas, five dysplastic epithelia, seven hyperplastic epithelia, and four normal epithelia were obtained from 21 hamster tongues by applying 1.0% acetone solution of DMBA on the left lingual mucosa after scratching with a root canal broach. Ultrastructural examination revealed that the number of microvilli increased, whereas that of desmosomes decreased during carcinogenesis. Cell proliferation was analyzed by means of 5-bromodeoxyuridine (BrdU) immunohistochemistry and in situ hybridization (ISH) for histone H3 mRNA. The BrdU and histone H3 mRNA labeling indices (LIs) were lowest for normal epithelium, higher for hyperplastic and dysplastic epithelia, and highest for squamous cell carcinoma. Cytoplasmic histone H3 mRNA and nuclear BrdU were localized in virtually identical areas of serial sections. The correlation coefficient for the relationship between these two LIs was 0.97 (P 0.001). These results suggest that the assessment of cell proliferation using H3 mRNA ISH will be a useful technique for investigating biological behavior during carcinogenesis.     

小鼠晶状体发育过程中细胞增殖及突触素蛋白的表达

目的 探讨小鼠晶状体细胞增殖、细胞周期蛋白（cyclin) D1和突触素（SYN）的表达情况。方法 各日龄小鼠共150只,HE染色和
4’6-二脒基-2-苯基吲哚（DAPI）染色,在光镜下观察小鼠晶状体的一般结构;用抗细胞增殖核抗原（PCNA）免疫荧光法以及5′-溴脱氧尿嘧
啶核苷（BrdU）技术标记晶状体增殖细胞;用免疫荧标记标价法检测细胞周期蛋白cyclin D1和SYN在晶状体的表达。结果 孕8d（E8）左右,
晶状体由最初的晶状体板分化而来,是由单层柱状上皮细胞围成的空泡,空泡再进一步分化,被原始晶状体纤维所替代,逐渐形成晶状体;
BrdU和PCNA的阳性细胞主要分布在晶状体上皮细胞的前囊中央区,但出生10d（P10）以后BrdU阳性细胞已不在晶状体上皮细胞分布,而PCNA持
续表达。Cyclin D1阳性细胞在胚胎期主要在晶状体上皮细胞的赤道部表达,出生早期在晶状体上皮细胞的前囊中央区,出生5d（P5）以后在赤
道前区分布,赤道部有少量阳性细胞;在晶状体发育过程中,SYN可以在晶状体基质中和赤道部胞体中发现。结论 细胞增殖对晶状体的发育和
修复起着重要的作用;cyclin D1可能参与调控赤道部细胞的消失,因此保持晶状体的透明度;SYN在晶状体的表达说明了它对晶状体的透明有
着重要的作用,通过调控钙离子内流入胞体和基质中。     

酪氨酸激酶受体KIT在小鼠结肠黏膜上皮的表达及其功能

目的 探讨酪氨酸激酶受体KIT在小鼠结肠黏膜上皮的表达及作用。 方法 应用Western blotting和RT-PCR技术检测c-kit基因及KIT
蛋白在野生型C57BL/6小鼠结肠黏膜上皮的表达;用免疫荧光染色显示野生型C57BL/6小鼠以及乙基亚硝酸钠（ENU）诱变的c-kit基因点突变纯
合子小鼠（Wads ^m/m）结肠黏膜上皮KIT阳性细胞的部位;腹腔注射5-溴脱氧尿嘧啶核苷（BrdU）（30 mg/kg）观察对比野生型以及Wads m/m
小鼠结肠黏膜上皮BrdU掺入情况及动态变化,分析KIT在结肠黏膜上皮更新过程的作用。 结果 RT-PCR检测结果表明,在野生型小鼠结肠黏膜组
织表达 c-kit基因, Western blotting证明在肠黏膜组织存在KIT蛋白;免疫荧光染色显示,KIT阳性细胞位于结肠黏膜上皮,主要位于肠腺隐
窝基底部,但是Wads ^m/m小鼠KIT阳性细胞数以及KIT蛋白表达量较野生型小鼠明显减少;BrdU掺入实验发现,Wads ^m/m小鼠远端结肠黏膜上皮
BrdU摄入和更新速度较野生型小鼠明显减慢。结论 结肠黏膜上皮表达KIT蛋白与结肠黏膜上皮的更新密切相关。     

The BrdU content of DNA is decreased during reversal of inhibition of myogenesis by deoxycytidine

The mechanism by which 5-bromodeoxyuridine (BrdU) inhibits cell differentiation is unresolved. The ability of deoxycytidine to reverse the inhibition of myogenesis produced by BrdU has been cited as evidence that the inhibition is not a direct result of the incorporation of BrdU into cellular DNA. In contrast to previous work, the present study demonstrates a direct correlation between the effects of deoxycytidine on myogenic cells and a reduction in the substitution of BrdU for thymidine in the DNA. Furthermore, the reversal occurs at the same degree of BrdU substitution (20–30%) as is required to inhibit myogenesis when cells are grown in BrdU alone or with deoxycytidine in a medium that prevents the conversion of deoxycytidine to thymidine. The effects of deoxycytidine thus do not support a mechanism of action of BrdU in myogenic cells independent of its effects on DNA.     

Molecular and cellular analysis of embryonic avian tongue development.

Signalling cascades first described in Drosophila have been found to regulate patterning and outgrowth in a number of structures in higher vertebrates. We sought to determine whether the evolutionarily conserved genes were important during the development of the tongue. In situ hybridisation was used to determine the temporo-spatial expression of a panel of conserved genes. Histological examination and incorporation of BrdU were used to determine the mechanism by which the tongue develops. We show that evolutionarily conserved genes were expressed in distinct dynamic patterns during tongue development. Sonic Hedgehog (Shh) and Patched (Ptc) were found only in the dorsal tongue epithelium. Shh expression was only observed in the suprabasal layers, whereas Ptc was observed in both basal and suprabasal layers. Cell division in the epithelium was concentrated in regions devoid of Shh. Expression of bone morphogenetic protein-7 (BMP) was identical to that of Shh. Shh and Ptc expression were never detected in the mesenchyme. Ectopic expression of Noggin (a potent antagonist of the BMPs) caused severe abnormalities in tongue morphology, including swelling of the mesenchymal component and a thickening of the epithelial layer. Data from this study suggests that the epithelium and mesenchyme express quite different genes during development. However BMP activity acts to inhibit growth in both tissues.