Loss of bcl-2 expression in ductal carcinoma in situ of the breast relates to poor histological differentiation and to expression of p53 and c-erbB-2 proteins

Aim : This study (1) investigates the incidence of bcl-2 protein expression in a series of 108 cases of ductal carcinoma in situ (DCIS), including 25 with early invasive carcinoma, and (2) evaluates the relationship of bcl-2 expression to the histological grade of DCIS and to the expression of oestrogen receptor (ER), c-erbB-2 and p53 proteins.  

Methods and results

The expression of bcl-2, oestrogen receptor (ER), c-erbB-2 and p53 proteins was determined immunohistochemically. Cases were regarded as positive for individual antibodies when at least 10% of the DCIS cells showed positive staining. DCIS was graded histologically as well ( n  = 9), intermediately ( n  = 24), or poorly differentiated ( n  = 75). bcl-2 expression was documented in 57 cases (53%) and was strongly associated with the histological grade of DCIS ( P  < 0.0001). All cases of well-differentiated DCIS were bcl-2 positive and loss of bcl-2 expression was almost exclusively confined to poorly differentiated DCIS lesions. bcl-2 expression was also closely associated with positive ER status ( P  < 0.0001). Forty-seven of 57 (82%) bcl-2 positive cases were ER positive while 49/51 (96%) bcl-2 negative cases were ER negative. There was a significant inverse correlation between bcl-2 expression and both p53 protein expression ( P  = 0.0004) and c-erbB-2 expression ( P  < 0.0001). Nineteen of 24 (79%) p53 positive cases and 38/45 (84%) c-erbB-2 positive cases showed loss of bcl-2.  

Conclusions

Loss of bcl-2 expression occurs in poorly differentiated DCIS and is related to negative ER status and to positive p53 and c-erbB-2 status. This pattern of bcl-2 expression and its association with other biological markers in DCIS is similar to that reported in invasive breast carcinoma.     

Commentary on the safety of red cells preserved in extended-storage media for neonatal transfusions

Red cells preserved in extended-storage media are the standard product dispensed by many regional blood centers. When the red cells are intended for neonatal transfusion, concern exists about the safety of the relatively high quantities of additives present in these media. Definitive studies to address these concerns are not available. Therefore, to estimate the effects of additives and to delineate circumstances in which they might be harmful, the quantities transfused in defined clinical settings were calculated, and the following recommendations are offered for transfusing infants less than 4 months of age. First, red cells preserved in extended-storage media should present no substantive risks when used for small-volume (approximately 10 mL/kg) transfusions of premature infants and can be used without additional processing. Second, the risks of the most premature neonatal patients or those with severe renal and/or hepatic insufficiency cannot be defined clearly, and, because data are not available to ensure safety for these infants, removal of the additive medium and resuspension of the red cells in saline or albumin solution immediately before transfusion are recommended. Third, following a similar rationale, it seems prudent to avoid using entire units of red cells preserved in extended-storage media in massive transfusion settings (e.g., exchange transfusion, cardiac surgery, and extracorporeal membrane oxygenation). In these settings, the preservative medium should be removed and the red cells resuspended in the fluid that is most appropriate for the procedure that is planned. It must be emphasized that these recommendations are based on calculations and hypothetical settings, not actual data. Accordingly, they are tentative and should be altered as definitive information becomes available.     

骨髓间充质干细胞的分化潜能及临床应用价值

目的:观察人的骨髓间充质干细胞多分化潜能及在糖尿病治疗领域的价值。方法:实验于2005-07/2006-07在青岛大学医学院附属医院内分泌科完成。骨髓来源于非造血系统疾病的16岁儿童胸骨骨髓血(查体以排除造血系统疾病,结果显示为健康体质),经得家属同意。Percoll淋巴细胞分层液分离骨髓间充质干细胞,取第3代细胞,等密度接种于培养瓶中,经CD44抗体、CD45抗体、CD34抗体鉴定。取其第4代细胞,诱导其向脂肪细胞及神经细胞分化,利用碱性成纤维细胞生长因子预处理先获得巢蛋白阳性细胞,分别用两种方法诱导其向胰岛祖细胞的转化:化学物质诱导和共培养法诱导,免疫荧光检测胰岛祖细胞标志-胰十二指肠同源异型盒基因(蛋白的表达,电化学发光法检测其是否表达胰岛素)。胶原酶消化法获取胰腺间充质干细胞,鉴定,用添加碱性成纤维细胞生长因子的无血清低糖DMEM使其增殖。将胰腺间充质干细胞接种于底层已接种骨髓间充质干细胞的6孔板共培养,共培养法诱导骨髓间充质干细胞向胰岛祖细胞的初步转化。结果:成脂诱导及成神经诱导可获得油红O染色阳性细胞以及巢蛋白阳性细胞。化学法向胰岛祖细胞诱导后可检测到PDX-1免疫反应阳性细胞。共培养法诱导也可获得PDX-1免疫反应阳性细胞。新生儿胰腺具有巢蛋白、CK-19阳性的胰腺间充质干细胞,体外高糖诱导可形成胰岛样细胞团。结论:骨髓间充质干细胞在体外具有诱导分化为脂肪细胞、神经元样细胞及胰岛祖细胞的潜能。新生儿胰腺间充质干细胞向胰岛细胞分化过程中所分泌的一些物质对骨髓间充质干细胞向胰岛祖细胞的转化具有促进作用。     

Pronounced cytostatic activity and bystander effect of a novel series of fluorescent tricyclic acyclovir and ganciclovir derivatives in herpes simplex virus thymidine kinase gene-transduced tumor cell lines

A number of tricyclic acyclovir (ACV) and ganciclovir (GCV) derivatives substituted with bulky lipophilic groups have been identified as potent and highly selective cytostatic agents against herpes simplex virus type 1 (HSV-1)-thymidine kinase (TK) gene-transduced human osteosarcoma and murine mammary carcinoma tumor cells. Although their affinity for HSV-1 TK was inferior to that of ACV or GCV, their cytostatic potency and selectivity was at least as high as observed for the parental ACV and GCV compounds. The tricyclic ACV and GCV derivatives were also endowed with a very pronounced bystander effect in cell culture, albeit at relatively high drug concentrations. Unlike ACV and GCV, the tricyclic purine derivatives are endowed with intrinsically strong fluorescent properties, which allow simple and sensitive monitoring of drug concentrations in biological fluids and tissues. Also, the lipophilicity of the tricyclic purine derivatives is much higher than that of ACV and GCV, and this may allow better uptake of these derivatives from the blood into the central nervous system.     

Akutversorgung und Spätfolgen bei kindlichen Verletzungen der Halswirbelsäule

Pediatric injuries in the region of the cervical spine are rare. Timely diagnostics and a rapid and consistent treatment are essential. This is probably the best initial situation to avoid delayed sequelae as these would mean lifelong impairment for the child. Therefore, this article deals with the acute treatment and delayed sequelae of pediatric injuries of the cervical spine.     

Inhibitory effects of diltiazem on platelet activation caused by ionophore A23187 plus ADP or epinephrine in subthreshold concentrations

We examined the effects of the slow channel Ca++ blocker diltiazem on human platelet aggregation and TXA2 generation. Diltiazem inhibited platelet aggregation induced by 2 microM ADP or 5.5 microM epinephrine alone at 5 and 50 micrograms/ml (11.1 and 111 microM), respectively, and that induced by threshold concentrations of ADP or epinephrine at 0.2 to 1.0 micrograms/ml (0.4 to 2.2 microM). Platelet TXA2 generation stimulated by either ADP or epinephrine alone was inhibited by diltiazem in concentrations above the clinically achieved range (0.05 to 0.2 micrograms/ml, 0.1 to 0.4 microM). When PRP was stimulated with subthreshold concentrations of the Ca++ ionophore A23187 followed by subthreshold concentrations of ADP or epinephrine, a marked potentiation of platelet aggregation and TXA2 generation was observed. Incubation of PRP with diltiazem in a pharmacologic range resulted in marked reduction in ionophore A23187-induced potentiation of platelet activity caused by ADP or epinephrine. In other experiments, diltiazem was found to have no effects on PGI2-induced platelet aggregation inhibition. On the basis of these data, we conclude that (1) Ca++ flux across the platelet membrane stimulates platelet activity of subthreshold concentrations of ADP or epinephrine and (2) diltiazem in therapeutic concentrations reduces platelet activation induced by ionophore A23187 plus ADP or epinephrine, most likely by inhibiting Ca++ flux. These effects of diltiazem may not be observed in the therapeutic range if aggregatory concentrations of ADP or epinephrine alone are used.     

Loss of fibroblast HIF-1α accelerates tumorigenesis

Solid tumors consist of malignant cells and associated stromal components, including fibroblastic cells that contribute to tumor growth and progression. Although tumor fibrosis and aberrant vascularization contribute to the hypoxia often found in advanced tumors, the contribution of hypoxic signaling within tumor-associated fibroblasts to tumorigenesis remains unknown. In this study, we used a fibroblast-specific promoter to create mice in which key hypoxia regulatory genes, including VHL, HIF-1α, HIF-2α, and VEGF-A, were knocked out specifically in tumor stromal fibroblasts. We found that loss of HIF-1α and its target gene VEGF-A accelerated tumor growth in murine model of mammary cancer. HIF-1α and VEGF-A loss also led to a reduction in vascular density and myeloid cell infiltration, which correlated with improved tumor perfusion. Together, our findings indicate that the fibroblast HIF-1α response is a critical component of tumor vascularization.