rHuG-CSF increases the platelet-neutrophil complex formation and neutrophil adhesion molecule expression in volunteer granulocyte and stem cell apheresis donors

Several reports have shown that granulocyte colony-stimulating factor (G-CSF) administration induces a transient, mild hypercoagulable state, which might predispose certain donors to thrombotic complications. In the present study, changes in the expression of neutrophil adhesion molecules (CD11b/CD18, CD62L) and platelet-neutrophil complex formation following rHuG-CSF administration were investigated in normal granulocyte and stem cell donors. For granulocyte apheresis (N = 10), rHuG-CSF (5 microg/kg) was given subcutaneously every 12 h three times and apheresis was carried out two hours after the last dose. For stem cell apheresis (N = 8), rHuG-CSF (10 microg/kg/day) was given subcutaneously for 5 days and apheresis was carried out when peripheral CD34+ cell counts exceeded 20 cell/microL. Expression of neutrophil adhesion molecules (CD11b/CD18, CD62L) and platelet-neutrophil complex formation following rHuG-CSF administration were investigated in donors by a flow cytometric method. A significant increase in neutrophil counts (P < 0.001), and decreases in platelet counts (P < 0.01) and hemoglobin levels (P < 0.01) occurred following G-CSF administration. The expression of CD11b/CD18 significantly increased (P < 0.001) over pretreatment values with G-CSF administration and returned to baseline 1 week after stopping the drug. In contrast, CD62L expression was decreased (P < 0.01) with G-CSF and returned to normal after cessation of the drug. rHuG-CSF caused more than a two-fold increase (from 0.3 to 7.0 x 10(9)/L) in circulating platelet-neutrophil complexes (P < 0.01), which returned to normal after 1 week. Although clinical significance of these laboratory changes is not clear, the occurrence of neutrophil activation and increased platelet-neutrophil complex formation might predispose certain donors or patients to thrombotic complications following G-CSF administration.     

转录因子FOXO1和干细胞标志物CD133与神经母细胞瘤临床病理因素相关性

目的探究叉头框转录因子O亚族(FOXO)1与干细胞标志物CD133在神经母细胞瘤(NB)中的表达及其相关性分析。方法使用免疫组织化学方法检测53例NB组织(病例组)和27例癌旁正常组织(对照组)中FOXO1和CD133蛋白表达情况;应用Western印迹检测3例NB组织及3例癌旁组织FOXO1和CD133蛋白表达情况。结果 FOXO1在病例组中阳性表达率显著低于对照组;CD133在病例组中阳性表达率显著高于对照组;FOXO1、CD133与临床分期、组织类型和病理分型存在相关性(P<0.05);FOXO1在NB组织显著低于癌旁组织,而CD133在NB组织显著高于癌旁组织;FOXO1和CD133在蛋白表达水平上有明显相关性(P<0.05)。结论 FOXO1可能通过负性调节CD133表达来调节NB干细胞特性,是NB发生发展的重要机制。     

肝富集转录因子对HBV基因转录和复制的调控作用

目的建立乙型肝炎病毒（HBV）在非肝源细胞的复制体系,探讨肝富集转录因子对HBV基因转录和复制的调控作用。方法用复制型HBV重组质粒pHBV4.1与肝富集转录因子HNF3、HNF4和RXRα/PPARα的表达质粒分别共转染非肝源细胞株NIH3T3。用Northern吸印杂交检测HBV 3.5 kb、2.4/2.1 kb、0.7 kb mRNA的转录情况,Southern吸印杂交检测HBV DNA复制中间体水平。结果复制型HBV重组质粒pHBV4.1在肝癌细胞中能检测到3.5 kb HBV RNA转录产物和HBV DNA复制中间体;用pHBV4.1转染NIH3T3细胞后,在没有肝富集转录因子表达质粒共转染时未能检出3.5 kb HBV RNA转录产物,也无HBV DNA复制中间体;当用肝富集转录因子共转染时,HNF4和RXRα/PPARα的表达能够激活3.5 kb HBV RNA转录和HBV DNA复制,而HNF3未能激活HBV复制,但在HNF4或RXRα/PPARα介导的病毒复制体系中,均见3.5 kb HBV mRNA的转录和HBV DNA复制水平下降。结论利用肝富集转录因子成功地建立HBV在非肝源细胞中的转录与复制。其中,HNF4和RXRα/PPARα可支持HBV在非肝源细胞中转录与复制;HNF3抑制HBV肝特异性基因转录与复制。     

Comparison of lung morphology in COPD secondary to cigarette and biomass smoke

OBJECTIVE: To compare lung morphology in chronic obstructive pulmonary disease (COPD) secondary to cigarette smoke (CS) and biomass smoke (BS). METHODS: Necropsies of women with COPD diagnosis by lung pathology and unique exposure to BS (n = 27) or CS (n = 21) matched by age and place of origin. Lungs were macroscopically and microscopically examined to evaluate the extent of emphysema, pigment deposition, and abnormalities in pulmonary arteries, large airways (including the Reid index) and small airways (SAWs) by a semiquantitative method. RESULTS: Both groups had variable degrees of emphysema and SAWs disease. Patients exposed to BS had more lung fibrosis and pigment deposition and thicker pulmonary arterial intima than smokers, who had more emphysema and epithelial damage (goblet cell metaplasia). The Reid index was similar in both groups. CONCLUSION: Lengthy exposure to BS can produce emphysema and other lesions typically observed in cigarette smokers, but with a slightly different distribution. Whether the differences observed are the consequence of severity of exposure or smoke composition, or both, remains to be clarified.     

Role of transcription factors CREB and CREM in cAMP-regulated transcription during spermatogenesis.

The cAMP response element binding protein (CREB) and the cAMP-responsive element modulator (CREM) are cyclically expressed at high levels during spermatogenesis. Cyclical expression of CREB and CREM in germ and somatic Sertoli cells correlates with the fluctuations in cAMP signaling induced by the pituitary gonadotropic hormones FSH and LH both during sexual maturation of the testis and during the 12-day cycles of spermatogenesis that occur in the adult testis. CREB and CREM are expressed at different times during the spermatogenic cycle, undergo programmed sequential switches from activator to repressor isoforms by mechanisms of alternative exon splicing and promoter usage, and are autoregulated by cAMP signaling in opposing directions. cAMP response elements located in the promoter of the CREB gene upregulate the expression of activator CREBs, whereas cAMP autoregulatory response elements in the internal promoter of the CREM gene induce expression of repressor CREM isoforms. The complex mechanisms for the regulation of the expression of CREB and CREM in the testis appear to reflect critical adaptations for regulating key target genes essential for the development of germ cells.     

Regulation of neutrophil and eosinophil secondary granule gene expression by transcription factors C/EBP epsilon and PU.1

In the bone marrow of C/EBP epsilon(-/-) mice, expression of neutrophil secondary and tertiary granule mRNAs is absent for lactoferrin (LF), neutrophil gelatinase (NG), murine cathelin-like protein (MCLP), and the cathelin B9; it is severely reduced for neutrophil collagenase (NC) and neutrophil gelatinase-associated lipocalin (NGAL). In addition, the expression of eosinophil granule genes, major basic protein (MBP), and eosinophil peroxidase (EPX) is absent. These mice express C/EBP alpha, C/EBP beta, and C/EBP delta in the bone marrow at levels similar to those of their wild-type counterparts, suggesting a lack of functional redundancy among the family in vivo. Stable inducible expression of C/EBP epsilon and C/EBP alpha in the murine fibroblast cell line NIH 3T3 activated expression of mRNAs for B9, MCLP, NC, and NGAL but not for LF. In transient transfections of C/EBP epsilon and C/EBP alpha, B9 was strongly induced with weaker induction of the other genes. C/EBP beta and C/EBP delta proteins weakly induced B9 expression, but C/EBP delta induced NC expression more efficiently than the other C/EBPs. The expression of MBP was inefficiently induced by C/EBP epsilon alone and weakly induced with C/EBP epsilon and GATA-1, but the addition of PU.1 resulted in a striking cooperative induction of MBP in NIH 3T3 cells. Mutation of a predicted PU.1 site in the human MBP promoter-luciferase reporter construct abrogated the response to PU.1. Gel-shift analysis demonstrated binding of PU.1 to this site. MBP and EPX mRNAs were absent in a PU.1-null myeloid cell line established from the embryonic liver of PU.1(-/-) mice. Restitution of PU.1 protein expression restored MBP and EPX protein expression. This study demonstrates that C/EBP epsilon is essential and sufficient for the expression of a particular subset of neutrophil secondary granule genes. Furthermore, it indicates the importance of PU.1 in the cooperative activation of eosinophil granule genes.     

Erythroid colony formation in primary proliferative polycythaemia,idiopathic erythrocytosis and secondary polycythaemia: sensitivity to erythropoietic stimulating factors

Summary This is a study of in vitro production of erythropoietic colonies from peripheral blood and bone marrow of normal subjects and patients with different polycythaemic conditions. Proliferative stimuli included: 1 Fetal calf serum (FCS) only. 2 FCS plus a source of erythropoietic-stimulating activity (ESA). 3 FCS + ESA + erythropoietin (Ep). It was found that normal subjects and patients with secondary polycythaemia (SP) exhibited full colony growth only in the presence of both ESA and Ep, while patients with primary proliferative polycythaemia (PPP) showed colony production with FCS alone, further enhanced in the presence of ESA and Ep. A group of patients with idiopathic erythrocytosis (IE), namely with an increase of red cell mass not accompanied by other signs of myeloproliferative disorder, and without underlying cause, showed a heterogeneous response to ESA which in some patients was significantly greater than in normal subjects or in SP patients. It appears therefore that sensitivity of erythropoietic colony formation to Ep and ESA may be helpful in differentiating among various forms of polycythaemia; this study also establishes the heterogenicity of the IE group.