脱细胞羊皮基质的制备与性能检测

目的:制备一种廉价、安全而且来源丰富的生物敷料,以用于烧伤患者微粒皮移植后和削痂后创面的保护。方法:实验于2003-10/2007-03在积水潭医院烧伤研究所完成。①主要材料:健康山羊5只,纯种新西兰大白兔10只;戊二醛猪皮(清华源兴生物制品公司);冷冻保存的人皮(志愿者捐献)。②实验方法:山羊麻醉处死,去毛后取全层皮肤,通过反复冻融、配合超声震荡和洗脱液处理,在保证胶原支架完整情况下去除导致皮肤排斥反应的细胞成分,获得脱细胞羊皮基质,冷冻保存。③实验评估:利用常规病理检查和电子显微镜检查其有无明显细胞碎片残留;细菌真菌培养,测试其机械强度、亲水性和细胞毒性,通过动物埋藏实验确定其组织相容性,并与戊二醛交联的脱细胞猪皮以及冷冻保存的人皮作对照。结果:制备的脱细胞羊皮基质,常规病理检查、免疫组织化学染色、电子显微镜检查均未发现表皮和真皮细胞残留;细菌真菌培养呈阴性;为亲液固体,质感柔软有弹性,含水量、极限抗拉强度、应力-应变量、应力松弛特征、蠕变性等物理性能均与冷冻人皮相似,明显优于戊二醛猪皮;细胞毒性0级,在国家标准允许范围内;植入兔背部脊柱两侧的肌肉组织内后,能够与组织很好的融合,并诱导细胞和血管长入,组织相容性良好。结论:采用冻融法制备的脱细胞羊皮敷料安全、廉价、来源丰富,达到临床手术应用的保护性生物敷料标准。     

On and off-target effects of telomere uncapping G-quadruplex selective ligands based on pentacyclic acridinium salts

Quadruplexes DNA are present in telomeric DNA as well as in several cancer-related gene promoters and hence affect gene expression and subsequent biological processes. The conformations of G4 provide selective recognition sites for small molecules and thus these structures have become important drug-design targets for cancer treatment.The DNA G-quadruplex binding pentacyclic acridinium salt RHPS4 (1) has many pharmacological attributes of an ideal telomere-targeting agent but has undesirable off-target liabilities. Notably a cardiovascular effect was evident in a guinea pig model, manifested by a marked and sustained increase in QTcB interval. In accordance with this, significant interaction with the human recombinant β2 adrenergic receptor, and M1, M2 and M3 muscarinic receptors was observed, together with a high inhibition of the hERG tail current tested in a patch clamp assay.Two related pentacyclic structures, the acetylamines (2) and (3), both show a modest interaction with β2 adrenergic receptor, and do not significatively inhibit the hERG tail current while demonstrating potent telomere on-target properties comparing closely with 1. Of the two isomers, the 2-acetyl-aminopentacycle (2) more closely mimics the overall biological profile of 1 and this information will be used to guide further synthetic efforts to identify novel variants of this chemotype, to maximize on-target and minimize off-target activities.Consequently, the improvement of toxicological profile of these compounds could therefore lead to the obtainment of suitable molecules for clinical development offering new pharmacological strategies in cancer treatment.     

Near-triploid and near-tetraploid acute lymphoblastic leukemia of childhood

Cytogenetic and DNA flow cytometric analyses of leukemic cells from 1,971 children with newly diagnosed acute lymphoblastic leukemia (ALL) identified stem lines with modal chromosome numbers greater than 65 in 26 patients (1.3%). Near-triploidy (66 to 73 chromosomes) was found in six cases and near-tetraploidy (82 to 94 chromosomes) in 20. A striking morphologic finding was the presence of clumped chromatin with grooved nuclei or Rieder cell formation in 20 cases. Other than a slight excess of the pre-B immunophenotype, the near-triploid cases did not appear to differ substantially from the general ALL population in clinical features. In contrast, near-tetraploid cases were more often associated with a T-cell immunophenotype (47% v 14%, P less than .001) and L2 morphology (30% v 22%, P less than .01), and were older at diagnosis (median age, 8.6 v 4.8 years, P = .01) than cases with other ploidies. Moreover, an unusually high proportion of near-tetraploid cases tested (6 of 15) expressed one or more of the myeloid-associated antigens CD13, CD15, and CD33. Despite the use of contemporary intensive chemotherapy and short follow-up for most patients, 6 of the 20 near- tetraploid cases have relapsed or died. This study suggests that the near-tetraploid subtype differs from other cases of hyperdiploid greater than 50 ALL, which have been associated with a favorable prognosis.     

Platelets express a membrane protein complex immunologically related to the fibroblast fibronectin receptor and distinct from GPIIb/IIIa

We have previously identified and characterized a membrane glycoprotein complex (GP150/135) that functions as fibronectin receptor (FN-R) in fibroblast adhesion. Here we report that an immunologically related protein complex is expressed at the surface of human platelets. Antibodies monospecific for the smaller subunit (GP135) of the fibroblast FN-R in fact specifically stained the platelet surface, as determined by FACS analysis, and reacted with a component of molecular weight (mol wt) 138,000 as shown in western blots of platelet membranes. Moreover, the same antibodies precipitated the 138,000 component together with a 160,000 protein, suggesting that the two molecules are associated in a supramolecular complex. A comparative analysis indicated that this protein complex is distinct from the GPIIb/IIIa complex, known to function as a receptor of wide specificity for fibrinogen, fibronectin, and von Willebrand factor. Differential extraction experiments revealed that the platelet 138,000 component is an integral membrane protein.     

The Catalonian Expert Patient Programme for Chagas Disease: An Approach to Comprehensive Care Involving Affected Individuals

The Catalonian Expert Patient Programme on Chagas disease is a initiative, which is part of the Chronic Disease Programme. It aims to boost responsibility of patients for their own health and to promote self-care. The programme is based on nine sessions conducted by an expert patient. Evaluation was focusing in: habits and lifestyle/self-care, knowledge of disease, perception of health, self-esteem, participant satisfaction, and compliance with medical follow-up visits. Eighteen participants initiated the programme and 15 completed it. The participants were Bolivians. The 66.7 % of them had been diagnosed with chagas disease in Spain. The 100 % mentioned that they would participate in this activity again and would recommend it to family and friends. The knowledge about disease improve after sessions. The method used in the programme could serve as a key strategy in the field of comprehensive care for individuals with this disease.     

Effect of recombinant canine granulocyte-macrophage colony-stimulating factor on hematopoietic recovery after otherwise lethal total body irradiation

Recombinant canine granulocyte-macrophage colony-stimulating factor (rcGM-CSF) was studied in normal dogs and in dogs receiving otherwise lethal total body irradiation (TBI) without marrow transplant. Five normal dogs receiving 25 micrograms/kg of rcGM-CSF by subcutaneous (SC) injection twice daily (BID) for 14 days showed increases in peripheral blood neutrophil counts of three to five times the baseline. Platelet counts decreased during administration of rcGM-CSF to a mean nadir of 52,800. Ten dogs received 400 cGy TBI at 10 cGy/min from two opposing 60Co sources and no marrow graft. Within 2 hours of TBI, rcGM-CSF was begun at a dose of 50 micrograms/kg SC BID for 5 doses and then continued at 25 micrograms/kg SC BID for 21 days. Only 1 of the 10 dogs receiving rcGM-CSF survived with complete and sustained recovery of hematopoiesis. One of 13 historical control dogs survived after 400 cGy with no hematopoietic growth factor or marrow infusion. Results with rcGM-CSF were compared with previous and concurrent data with G-CSF studied in the same model. Of 10 dogs receiving G-CSF, 8 survived with complete and sustained hematopoietic recovery, a significantly better survival than that seen with rcGM-CSF (P = .006). Neutrophil counts were sustained at higher levels after TBI for the first 18 days in the G-CSF group (P < .016) and the neutrophil nadirs were higher. No differences in neutrophil nadirs were noted between the rcGM-CSF and control groups. Dogs treated with rcGM-CSF experienced a more rapid decline of platelet counts than G-CSF-treated or control dogs over the first 18 days (P < .001). The nadir of the platelet count was higher in the control group than in either the G-CSF or rcGM-CSF group and no significant difference was observed between the G-CSF and rcGM-CSF groups. After otherwise lethal TBI (400 cGy) in dogs, rcGM-CSF was not effective in promoting hematopoietic recovery or improving survival.     

Temperature, heart rate, and blood pressure changes associated with clinical MR imaging at 1.5 T

Temperature, heart rate, and blood pressure responses to high-field-strength magnetic resonance (MR) imaging were studied in 50 patients who underwent procedures at exposures to radiofrequency radiation above the present recommended whole-body average specific absorption rate (SAR) of 0.4 W/kg. Body temperature significantly increased an average of 0.2 degrees C. The highest body temperature recorded after MR imaging was 37.5 degrees C. There was no significant correlation between the change (before and after imaging) in body temperature and whole-body average SARs. Changes in skin temperatures were variable, depending on anatomic site. The largest change was 3.5 degrees C, and the highest skin temperature recorded after imaging was 35.1 degrees C. There was a modest correlation between the change in skin temperatures and whole-body average SARs. Average heart rate and average mean blood pressure measured immediately before imaging were not significantly different afterward. High-field-strength MR imaging at the whole-body average SARs of 0.42-1.2 W/kg studied was not associated with any temperature- or hemodynamic-related deleterious effects.