肝外胆管癌患者癌组织与血清中PLK1、Aurora A水平的变化及其临床意义

目的:探讨有丝分裂调控酶polo样激酶1(PLK1)和Aurora A在肝外胆管癌患者癌组织及血清中的水平以及两者的临床意义。方法:用免疫组化法测定54例肝外胆管癌组织及20例癌旁胆管组织中PLK1和Aurora A的表达,用ELISA法检测25例肝外胆管癌患者与15例健康人血清中PLK1和Aurora A的浓度。分析两者表达与患者临床病理因素的关系,以及患者手术前后两者水平的变化。结果:肝外胆管癌患者中,癌组织中PLK1和Aurora A的阳性表达率均明显高于癌旁胆管组织(66.7%vs.25.0%;63.0%vs.15.0%,均P0.05),且PLK1和Aurora A的表达与肿瘤的分化程度、TNM分期和淋巴结转移密切有关(均P0.05);术前血清中PLK1和Aurora A的浓度均明显高于健康人群(434.85 pg/mL vs.256.00 pg/m L;644.64 pg/m L vs.375.73 pg/m L,均P0.05),术后两者水平均明显降低(均P0.05);PLK1与Aurora A在肝外胆管癌患者癌组织中的表达量与血清中浓度具有一致性(r=0.55;r=0.64,均P0.05);无论癌组织或血清中,PLK1与Aurora A水平均呈正相关(癌组织:r=0.47,P0.01;血清:r=0.71,P0.01)。结论:PLK1与Aurora A水平在肝外胆管癌患者癌组织与血清均升高,且两者水平的升高与肝外胆管癌的恶性进展密切相关;两者联合检测对肝外胆管癌早期诊断、治疗效果及预后的判断有一定价值。     

50例胃肠道平滑肌肿瘤临床病理研究

 本文报告50例胃肠道平滑肌肿瘤，胃部20例、十二指肠5例、空肠10例、回肠6例、结肠9例。良性29例、恶性21例。临床上本瘤诊断率低，当出现消化道出血、腹部肿块和腹痛时应高度怀疑本病。提出病理论断平滑肌肉瘤标准：1.瘤细胞核分裂≥1/10个高倍视野（HPF）、胃为3/10HPF.2.幼稚瘤细胞。3.瘤体直径大于7cm.4.瘤细胞对周围组织浸润。5.瘤细胞凝固性坏死。有第1项，有或无其他项；或后4项有其中两项即可考虑为平滑肌肉瘤，项目越多恶性程度越高。     

Postnatal maturation of the rabbit cortical collecting duct

The mature, fully differentiated cortical collecting duct plays a major role in the final renal regulation of Na⁺, K⁺ and H⁺ transport. To characterize the growth of this segment, we measured the outer diameter and the dry weight of cortical collecting ducts isolated from newborn, 1-month-old, and adult rabbits. During the 1st month of life no significant changes were observed; however, there was a 60% increase in both parameters after the 4th week of life. Growth-related accretion of K⁺ was demonstrated by showing tubular K⁺ content to increase by 60% with maturation. Concomitant with the increase in tubular size, total cell number per millimeter of tubular length rose by 30%. Approximately 50% of the observed increment in tubular size could be accounted for by cell hyperplasia, with the remaining increase resulting from cell hypertrophy. Hypertrophy of principal cells was confirmed by scanning electron microscopy, which demonstrated a doubling of the circumferential width without any change in longitudinal length. Hyperplasia was confirmed, using a fluorescent chromatin stain, by our finding of a mitotic frequency of 3/1000 cells in the neonatal mid-cortical collecting duct; the observed number of mitoses was 10-fold higher at the most cortical end (ampulla). The number of intercalated cells per millimeter of tubule length, identified by bright green fluorescence after cortical collecting ducts were stained with 6-carboxyfluorescein diacetate, was found to double during maturation, the increase being significant only after the 4th postnatal week. We conclude that maturation of the mid-cortical collecting duct results from both cellular hyperplasia and hypertrophy. It is unlikely that this segment plays a major role in regulating Na⁺, K⁺, and H⁺ transport in the neonatal kidney.     

Patterns of glial proliferation during formation of olfactory glomeruli in an insect

Partitioning of the first-order olfactory neuropil into glomeruli in the developing brain of the moth Manduca sexta occurs only in the presence of olfactory sensory axons and appears to be mediated by changes in glial cells (Oland et al.: J. Neurosci., 8:353-367, 1988). The arrival of sensory axons in the brain triggers changes in glial shape and position that lead to the formation of a glial scaffolding for the developing glomeruli. The presence of mitotic figures in glial cells at stages before glomeruli emerge (Oland and Tolbert: J. Comp. Neurol., 255:196-207, 1987) suggested that glial proliferation might also contribute to the formation of the glomerular envelopes. To determine whether glial proliferation is induced by olfactory axons, we have used 3H-thymidine to label dividing cells before, during, and after the formation of glomeruli and have compared the patterns of proliferation in normal and chronically unafferented olfactory neuropils. We found significant differences in mitotic indices only after glomerular walls had been established, indicating that the sensory axons induce the formation of glomerular envelopes primarily via the changes in glial morphology and distribution, not by stimulating glial proliferation.     

Fibrinogen/fibrin in atherogenesis

Fibrin is a major component of many atherosclerotic plaques. Within the intima there is continuous formation of fibrin, and continuous fibrinolysis. In aortic lesions, a lipoprotein bound to fibrin can be released by incubation with plasmin. Most of this lipoprotein is accounted for by Lp(a). The atherogenicity of Lp(a) may be more associated with lipid deposition than with inhibition of fibrinolysis. Fibrin degradation products may be chemotactic to monocyte-macrophages and stimulate smooth muscle cell proliferation.Corresponding author     

Induction by cyproterone acetate of DNA synthesis and mitosis in primary cultures of adult rat hepatocytes in serum free medium

The aim of this study was to elucidate whether serum-free conditions could be found in primary hepatocyte cultures under which the growth inducing properties of xenobiotics and hormones could be tested. Cyproterone acetate (CPA), a steroid with anti-androgenic and progestogenic activity, was chosen as a model compound because of its known strong mitogenic properties in rat liver in vivo. EGF served as a positive control. Induction of DNA synthesis was studied by [³H]-thymidine labeling and autoradiography. Mitoses were counted in hematoxylin stained specimens. The main steps which led to an efficient stimulation of DNA synthesis by CPA were (i) reduction of hormone concentrations to levels approaching (approx. 10 x) physiological concentrations better than the previously used pharmacological ones (up to 2500 x); (ii) supplementation with glucocorticoid (most effective at 10–100 nM dexamethasone); (iii) selection of the interval for cumulative labeling with thymidine at 44–68 h; (iv) lowering of cell density at seeding to 50 000 cells/cm² (subconfluency); (v) treatment with concentrations of 10–100 M CPA. With these conditions CPA labelling was 13–20% (increase 4-to 9-fold). Mitotic incidence was 0.56% (CPA) versus 0.08% in controls. From a dose response study (0.1–100 M) a no-effect-level for induction of DNA synthesis was found in the range of 0.1–1 M. None of the high concentrations of CPA did cause cytotoxicity as estimated by morphological observations or release of lactate dehydrogenase into the medium. This work demonstrates that CPA under appropriate, defined culture conditions induces DNA synthesis and mitosis. The findings suggest that CPA action on the liver in vivo is at least in part a direct one. The extent of induction was in the same range as found in vivo. The effective concentrations were almost equivalent to CPA liver concentrations after treatment in vivo. Thus the present culture conditions appear to simulate quite well in vivo liver growth control mechanisms. This stresses the versatility of primary hepatocyte cultures as a tool in toxicology for the study of normal and drug-induced growth responses.Abbreviations CPA cyproterone acetate - DEX dexamethasone - EGF epidermal growth factor - LI labeling index     

Synergism between inhibitors of Aurora A and KIF11 overcomes KIF15‐dependent drug resistance

The mitotic kinesin KIF11 (also called Eg5) plays critical roles in spindle functions. Although a number of small‐molecule inhibitors of KIF11 are currently in clinical development, drug‐resistance could be developed through compensation by another kinesin called KIF15. Using a newly developed infrared‐based cell system, we discovered that the effectiveness of one of the latest generations of KIF11 inhibitor (SB743921) could be enhanced with several inhibitors of Aurora A kinase. Evidence including live‐cell imaging and isobologram analysis indicated that targeting KIF11 and Aurora A together promoted monoastral spindle formation and mitotic catastrophe synergistically, supporting a model of parallel pathways of centrosome regulation by Aurora A and KIF11. We also developed a KIF15‐dependent SB743921‐resistance cell model. Significantly, the drug‐resistance could also be overcome with Aurora A inhibitors. These results provide a molecular basis for increasing the effectiveness of Aurora A and KIF11 inhibitors and tackling problems of drug resistance.     

Mutations in the human kinesin Eg5 that confer resistance to monastrol and S-trityl-l-cysteine in tumor derived cell lines

The kinesin Eg5 plays an essential role in bipolar spindle formation. A variety of structurally diverse inhibitors of the human kinesin Eg5, including monastrol and STLC, share the same binding pocket on Eg5, composed by helix α2/loop L5, and helix α3 of the Eg5 motor domain. Previous biochemical analysis in the inhibitor binding pocket of Eg5 identified key residues in the inhibitor binding pocket of Eg5 that in the presence of either monastrol or STLC exhibited ATPase activities similar to the untreated wild type Eg5. Here we evaluated the ability of full-length human Eg5 carrying point mutations in the drug binding pocket to confer resistance in HeLa and U2OS cells to either monastrol or STLC, as measured by the formation of bipolar spindles. Both transfected cells expressing wild type Eg5 and untransfected cells were equally sensitive to both inhibitors. Expression of Eg5 single point mutants R119A, D130A, L132A, I136A, L214A and E215A conferred significant resistance to monastrol. Certain mutations inducing monastrol resistance such as R119A, D130A and L214A also conferred significant resistance to STLC. For the first time at a cellular level, the propensity of selected Eg5 point mutants to confer drug resistance confirms the target specificity of monastrol and STLC for Eg5. These data also suggest a possible mechanism by which drug resistance may occur in tumors treated with agents targeting Eg5.     

十二指肠间质瘤临床分析30例

目的：分析十二指肠间质瘤的临床特征并探讨其外科治疗及预后因素．方法：回顾性分析我科1993-2006年30例十二指肠间质瘤的临床资料．运用SPSS统计软件进行生存分析,采用Kaplan Meier法和log-rank检验进行单因素生存预后分析,多因素分析采用Cox模型．结果：十二指肠间质瘤最常见症状为右上腹隐痛不适、消化道出血,发病部位以十二指肠降段多见．全组1,3,5 a生存率分别为92．95％,84．87％,69．15％．单因素分析Fletcher风险分级(X~2=10．570,P=0．0143),肿瘤大小(X~2=7．883,P=0．0485),核分裂象(＞5,≤5：X~2=14．155,P=0．0002),是否伴发远处转移(X~2=22．519,P=0．0000)具有显著性,是否行根治术(X~2= 17．8180,P=0．0000)比较差异具有显著性,多因素分析表明核分裂象是独立的预后影响因素．结论：十二指肠间质瘤宜采用局部切除为主的根治性手术,本组病例核分裂象是判断预后的独立因素．     

Update to the College of American Pathologists Reporting on Thyroid Carcinomas

Background The reporting of thyroid carcinomas follows the recommendations of the College of American Pathologists (CAP) protocols and includes papillary carcinoma, follicular carcinoma, anaplastic carcinoma and medullary carcinoma. Despite past and recent efforts, there are a number of controversial issues in the classification and diagnosis of thyroid carcinomas (TC) that, potentially impact on therapy and prognosis of patients with TC. Discussion The most updated version of the CAP thyroid cancer protocol incorporates recent changes in histologic classification as well as changes in the staging of thyroid cancers as per the updated American Joint Commission on Cancer staging manual. Among the more contentious issues in the pathology of thyroid carcinoma include the defining criteria for tumor invasiveness. While there are defined criteria for invasion, there is not universal agreement in what constitutes capsular invasion, angioinvasion and extrathyroidal invasion. Irrespective of the discrepant views on invasion, pathologists should report on the presence and extent (focal, widely) of capsular invasion, angioinvasion and extrathyroidal extension. These findings assist clinicians in their assessment of the recurrence risk and potential for metastatic disease. It is beyond the scope of this paper to detail the entire CAP protocol for thyroid carcinomas; rather, this paper addresses some of the more problematic issues confronting pathologists in their assessment and reporting of thyroid carcinomas. Conclusion The new CAP protocol for reporting of thyroid carcinomas is a step toward improving the clinical value of the histopathologic reporting of TC. Large meticulous clinico-pathologic and molecular studies with long term follow up are still needed in order to increase the impact of microscopic examination on the prognosis and management of TC.