Common patterns of increased and decreased Fos expression in midbrain and pons evoked by noxious deep somatic and noxious visceral manipulations in the rat

Immunohistochemical detection of the protein product (Fos) of the c-fos immediate early gene was used to study neuronal activation in the rostral pons and midbrain of halothane-anesthetised rats following noxious deep somatic or noxious visceral stimulation. In animals exposed only to halothane anesthesia. Fos-like immunoreactive (IR) neurons were located in the midbrain periaqueductal gray matter, tectum, and parabrachial nucleus. Following noxious stimulation of hindlimb muscle, knee joint, vagal cardiopulmonary, or peritoneal nociceptors, there was, compared to halothane-only animals, a significant increase in the numbers of Fos-like (IR) cells in the caudal ventrolateral periaqueductal gray and the intermediate gray lamina of the superior colliculus. Given the general agreement that increased Fos expression is a consequence of increased neuronal activity, the finding that a range of noxious deep somatic and noxious visceral stimuli evoked increased neuronal activity in a discrete, caudal ventrolateral periaqueductal gray region is consistent with previous suggestions that this region is an integrator of deep noxious evoked reactions. The noxious deep somatic and noxious visceral manipulations also evoked, compared to halothane-only animals, reductions in the numbers of Fos-like IR cells in the stratum opticum of the superior colliculus and the unlaminated portion of the external subnucleus of the inferior colliculus. To our knowledge this is the first report of reductions in Fos-expression in the tectum evoked by noxious stimulation. In separate experiments, the effects of noxious deep somatic and noxious visceral manipulations on arterial pressure and heart rate were measured. The noxious visceral manipulations evoked substantial and sustained falls in arterial pressure (15–45 mmHg), and heart rate (75–100 bpm), whereas the depressor and bradycardiac effects of the noxious deep somatic manipulations were weaker, not as sustained, or entirely absent. As similar distributions and numbers of both increased and decreased Fos-like IR cells were observed after each of the deep noxious manipulations, it follows that the deep noxious evoked increases and decreases in Fos expression were not secondary to the evoked depressor or bradycardiac effects. © 1996 Wiley-Liss, Inc.     

Detection of RET mutations in multiple endocrine neoplasia type 2a and familial medullary thyroid carcinoma by denaturing gradient gel electrophoresis

Germline missense mutations within the coding region of the RET proto-oncogene have recently been described in patients with the dominantly inherited cancer syndromes, multiple endocrine neoplasia type 2a (MEN 2a) and familial medullary thyroid carcinoma (FMTC). To date, the sequence variations occur in RET exons 10 and 11 and alter highly conserved cysteine residues in the proposed extracellular domain at codons 609, 611, 618, 620, and 634. To expedite rapid screening of populations at risk of MEN 2a or FMTC, we developed a PCR-based denaturing gradient gel electrophoresis (DGGE) strategy that detects polymorphisms occurring at all five Cys codons in both RET exons using identical gel conditions. In this report, the screening results from DGGE analysis of 15 distinct MEN 2a and FMTC mutations are shown. Each mutation generated a clearly distinguishable and unique homo- and heteroduplex band pattern. Given the highly efficient, reproducible, and sensitive nature of this approach, DGGE is particularly appropriate for rapid, large-scale screening of patients. Since prior knowledge of the RET mutation is unnecessary for analysis, DGGE is potentially valuable for distinguishing germline from seemingly sporadic medullary thyroid cancer as well as identifying novel sequence changes. © 1996 Wiley-Liss, Inc.     

Alteration of c-fos gene methylation in human gliomas

In an attempt to find a common DNA alteration occurring in human glioma, we examined DNA methylation in 34 gliomas of various pathological grades and compared them with those in normal cerebral subcortex DNA. The total methylated cytosine levels in the genome did not differ appreciably between the tumors and the normal tissues; however, the degree of DNA methylation in several proto-oncogenes and suppressor oncogenes showed some alterations. Among them, the c-fos gene demonstrated deviation from that of normal tissues in all cases examined, suggesting that the alteration of c-fos gene methylation plays a role in the early steps of human glioma development. © 1996 Wiley-Liss, Inc.     

Sporadic type composite pheochromocytoma with neuroblastoma: Clinicomorphologic, DNA content, and ret gene analysis

Composite pheochromocytomas (CP) account for only 3% of all pheochromocytomas. We analyzed the clinical, immunohistochemical, ultrastructural, DNA content, and a 634 ret mutation feature in a 56-yr-old Mexican woman with a CP localized in the right adrenal gland and associated with a blood pressure of 140/90 mmHg. Clinical symptoms were absent after surgery. The tumor showed pheochromocytoma and neuroblastoma components. This dual phenotype was supported by light microscopy and corroborated by immunohistochemistry and ultrastructural findings. Flow cytometric analysis showed that both components were diploid. A genetic mutational analysis of the ret oncogene in exon 11 showed no 634 mutation. This case demonstrates the indolent behavior of neuroblastoma associated with a sporadic-type CP in an adult patient.     

屈洛昔芬对妊娠大鼠黄体细胞凋亡和C-myc、Bax、Bcl-2蛋白表达的影响(英文)

目的:观察屈洛昔芬对妊娠大鼠黄体细胞凋亡的影响,并分析黄体中C-myc,Bax和Bcl-2蛋白表达与屈洛昔芬所诱导的黄体细胞凋亡间的可能联系。方法:大鼠于妊娠第2天口服给予屈洛昔芬20mg·kg~(-1)后,分别于第4天和第8天取卵巢,HE染色和TUNEL法检测黄体中凋亡细胞的存在,免疫组织化学方法观察黄体中C-myc,Bax和Bcl-2蛋白的表达,同时测定卵巢重量、蛋白质含量及血中孕酮水平。结果:大鼠经屈洛昔芬处理后,第4天卵巢黄体中出现明显的凋亡细胞,第8天时更加明显。卵巢重量、蛋白质含量及血清孕酮水平在第8天时显著下降。屈洛昔芬处理组大鼠卵巢黄体中C-myc蛋白表达在第4天即显著增加,Bax蛋白表达的显著增加在第8天可观察到,而Bcl-2蛋白在卵巢黄体中的表达无明显改变。结论:屈洛昔芬可诱导妊娠大鼠着床前黄体细胞凋亡,C-myc蛋白及Bax/Bcl-2蛋白表达的增加可能与该过程有关。     

雷公藤内酯抑制内皮细胞血管内皮生长因子表达与合成

目的:研究雷公藤内酯对血管内皮细胞生长因子(VEGF)mRNA表达及VEGF合成与分泌的影响,进一步探讨雷公藤内酯降低肾小球肾炎患者尿蛋白的作用机制。方法:以人内皮细胞系ECV-304为研究对象,利用半定量逆转录聚合酶链反应(RT-PCR),流式细胞仪,酶联免疫吸附法(ELISA)检测不同剂量雷公藤内酯对佛波脂(TPA)诱导的内皮细胞VEGFmRNA表达及VEGF合成与分泌的影响,用RT-PCR检测雷公藤内酯对内皮细胞c-fos/c-jun mRNA表达的影响。结果:TPA能够明显上调VEGF mRNA表达,蛋白合成与分泌。而雷公藤内酯可以抑制TPA诱导的内皮细胞VEGF mRNA表达及VEGF蛋白合成与分泌,该作用在10μg·L~(-1)时更为明显。同样,雷公藤内酯剂量依赖性地抑制TPA诱导的内皮细胞c-fos/c-jun mRNA的表达。结论:雷公藤内酯通过影响c-fos/c-jun基因转录而抑制内皮细胞VEGFmRNA表达及VEGF合成与分泌是雷公藤内酯降低肾小球肾炎患者尿蛋白的作用机制之一。     

多囊卵巢综合征中卵巢初级小卵泡bcl-2、bax的表达

目的 :探讨凋亡调节蛋白 bcl-2及 bax在多囊卵巢综合征 ( PCOS)患者的卵泡选择、发育的作用。方法 :1 8例 PCOS患者及 1 3名正常人 2 40个小卵泡按直径分为 4个组 ,采用免疫组织化学方法及 Konton Ibas灰度值的定量测定 ,用 SPSS软件统计 ,定量分析凋亡调节蛋白 bcl-2及 bax在 PCOS各级初级小卵泡中的表达。结果 :PCOS组各级窦前卵泡组间 bax蛋白表达有显著差异 ,随着卵泡直径的增加 bax表达明显增多 ,但直径 >1 2 0 μm的卵泡 bax表达减少 ,bcl-2 / bax灰度比值显著变小。 bcl-2蛋白表达相对稳定 :PCOS组直径在 60～ 1 2 0μm的卵泡 bcl-2表达显著高于正常对照组 ( P<0 .0 5 )。结论 :正常人及 PCOS卵巢组织各级窦前卵泡和窦卵泡均表达 bcl-2及 bax,随着卵泡的增大 ,bcl-2的表达增加 ,当卵泡直径 >1 2 0 μm时 ,细胞凋亡减弱 ,卵泡的增长加速。PCOS患者卵巢中各级窦前卵泡和窦卵泡凋亡蛋白 bcl-2及 bax的表达是正常的。     

HER-2/neu基因表达与卵巢上皮性癌的预后

【目的】探讨HER-2／neu基因在卵巢上皮性癌中的表达水平及其与预后的关系。【方法】利用免疫组化检测93例卵巢上皮性癌组织中HER-2／neu基因表达水平,并结合临床病理及随访资料进行分析。【结果】HER-2／neu基因主要为细胞膜及胞浆表达,阳性率为44．1％。其表达与临床分期、病理类型、组织学分级及残余灶直径等无相关性;但与预后相关。HER-2／neu表达阴性组的1,3,5年生存率分别为90．4％、74．0％、54．6％,弱阳性组分别为81．1％、45．4％、27．9％,强阳性组分别为80．0％、20．0％、10．0％。差异有显著性(P=0．0028)。COX模型多因素分析提示：HER-2／neu表达水平是影响卵巢癌预后的独立因素(P=0．005)。【结论】HER-2／neu基因表达水平可作为预测卵巢上皮性癌预后的可靠指标。     

Novel point mutation in exon 10 of the RET proto-oncogene in a family with medullary thyroid carcinoma

Medullary thyroid carcinoma (MTC) may occur sporadically or as part of the autosomal dominant multiple endocrine neoplasia type 2 (MEN 2). Three hereditary forms of MEN 2 have been identified: MEN 2A, MEN 2B, and familial MTC (FMTC). Missense germ-line mutations in the RET proto-oncogene have been identified as cause of these endocrine diseases. Mutations are found in exons 10 and 11 in MEN 2A and FMTC families and in a small number of families in exons 13, 14, and 15. Although a strong correlation between codon mutations and phenotypes has been described, not all the expected cystein codon mutations have been found. Therefore, the more mutations are found, the better it is possible to establish phenotype-genotype correlations. We report on a novel RET mutation at codon 611 in a family with MTC without other clinical manifestations and of rather benign course. Am. J. Med. Genet. 78:271–273, 1998. © 1998 Wiley-Liss, Inc.     

Src蛋白在肺腺癌A549细胞增殖浸润中的作用

目的:研究Src蛋白在肺腺癌A549细胞增殖浸润中的作用。方法:Westernblot检测Src蛋白在A549细胞的表达和磷酸化,MTT和Boydenchamber法分别检测抑制Src蛋白酪氨酸激酶对A549细胞体外增生和游走浸润的影响。结果:抑制Src蛋白酪氨酸激酶磷酸化,能够显著抑制A549细胞体外增生(P<0·01)和游走浸润(P<0·001)。结论:Src蛋白在肺腺癌A549细胞增殖浸润中发挥着重要作用,可能成为治疗肺腺癌的重要靶分子。