Effets de différents antioxydants sur la lipoperoxydation in vitro initiée par le radical °OH

Rationale and ObjectiveThe study of the oxidative stress remains an analytical challenge and a model of lipid peroxidation is described for this purpose: it is based on a hydrogen peroxide- porphyrin mix as an °OH generator and artificial «plasma» as acceptor.Materials and MethodsLipid peroxidation is assessed by the measurement of true malondialdehyde with a specific HPLC-UV method. This model is used for checking the role of four antioxidants, the first one being a hydrophilic compound (Trolox) and the others natural lipophilic molecules (α tocophérol, β carotene and lycopene).ResultsA linear relationship between concentrations and inhibition of MDA production is shown for the four compounds.ConclusionThis model, easy to handle, could therefore be useful for testing in vitro both hydrophilic and lipophilic antioxidants.     

Acquired non hypertrophic pyloric stenosis in children]

Pediatric non hypertrophic pyloric stenosis (NHPS) are uncommon. Their causes and treatments are debated.Material and methodRetrospective review of all cases of NHPS from 3 pediatric surgery services during the period 1984–2002.ResultsSix children, aged 17 months to 15 years, underwent surgery for NHPS. Clinical symptoms, food vomiting and loss of weight, were present for several weeks before the diagnosis of NHPS was made. The diagnosis was peptic stenosis in 3 cases and has not been established in 3 cases. Search for Helicobacter pylori was negative in all cases. Failure of specific medical treatment and endoscopic dilatations led to pyloric resection in 3 cases and pyloroplasty in 3 cases. Post operative course was uneventful with normal oral feeding and normalisation of weight status. Histologic data were aspecific. No recurrence was observed.DiscussionWe discuss the origin of the pyloric stenosis, regarding clinical, operative and pathological data: were the stenosis the cause or consequence of peptic ulcer? Peptic disease is always advocated, but difficult to prove and may be excessively incriminated. Late symptomatic congenital and acquired idiopathic pyloric stenosis should be recalled. In all cases of proved pyloric stenosis, after failure of medical and endoscopic treatment, a simple surgical procedure (pyloroplasty) associated with medical treatment seems to be effective.ConclusionThe diagnosis of NHPS should be suspected in a child with food vomiting and loss of weight if his age is not concordant with hypertrophic pyloric stenosis. Upper gastro-intestinal series and endoscopy are diagnostic. The precise cause of the stenosis is more difficult to asses. When the medical treatment fails, a pyloroplasty is usually curative.     

Region-specific growth properties and trophic requirements of brain- and spinal cord-derived rat embryonic neural precursor cells

To determine whether neural precursor cells have region-specific growth properties, we compared the proliferation, mitogenicity, and differentiation of these cells isolated from the embryonic day 16 rat forebrain and spinal cord. Neural precursor cells isolated from both regions were cultured in growth medium supplemented with epidermal growth factor, basic fibroblast growth factor, or epidermal growth factor+basic fibroblast growth factor. Under all three conditions, both neural precursor cell populations proliferated for multiple passages. While spinal cord-derived neural precursor cells proliferated moderately faster in epidermal growth factor-enriched growth medium, brain-derived cells proliferated much faster in basic fibroblast growth factor-enriched growth medium. When exposed to both epidermal growth factor and basic fibroblast growth factor, the two neural precursor cell populations expanded and proliferated more rapidly than when exposed to a single factor, with brain-derived neural precursor cells expanding significantly faster than spinal cord-derived ones (P<0.0001). Differentiation studies showed that both neural precursor cell populations were multi-potent giving rise to neurons, astrocytes, and oligodendrocytes. However, neuronal differentiation from brain-derived neural precursor cells was greater than spinal cord-derived ones (11.95+/-5.00% vs 1.92+/-1.13%; passage 2). Further, the two neural precursor cell populations differentiated into a similar percentage of oligodendrocytes (brain: 8.66+/-5.85%; spinal cord: 7.69+/-3.91%; passage 2). Immunofluorescence and Western blot studies showed that neural precursor cells derived from both regions expressed receptors for basic fibroblast growth factor and epidermal growth factor. However, brain-derived neural precursor cells expressed higher levels of the two receptors than spinal cord-derived ones in growth medium containing epidermal growth factor+basic fibroblast growth factor. Thus, our results showed that neural precursor cells isolated from the two regions of the CNS have distinct properties and growth requirements. Identifying phenotypic differences between these neural precursor cell populations and their growth requirements should provide new insights into the development of cell therapies for region-specific neurological degenerative diseases.     

CYP17A1基因新突变致17α-羟化酶/17,20-裂解酶部分性缺陷症

目的研究1例17α-羟化酶/17,20-裂解酶部分性联合缺陷症患者CYP17A1基因突变特点,并结合患者的临床表现与基因突变类型初步探讨P450C17酶蛋白的结构与功能的关系。方法收集1例17α-羟化酶/17,20-裂解酶部分性联合缺陷症患者的临床资料及其亲属血标本,提取基因组DNA,设计7对引物扩增CYP17A1基因的8个外显子及外显子与内含子的连接区域,琼脂糖凝胶电泳鉴定PCR产物,产物胶回收后直接做为DNA双链模板测序。DNA双链模板不一致的PCR产物经克隆后测序。测序结果在核苷酸序列数据库进行比较分析。结果患者CYP17A1基因突变检测结果为5994-5995delAT/7541C>T复合杂合子。这两种突变均未见报道。推测5994-5995delAT导致I259H,274X,突变形成的截短蛋白质缺少血红素结合区域,因此是没有功能的;而通过人类P450C17酶计算机模型分析显示7541C>T导致的A398V远离酶的活性中心,推测突变可能使酶的活性减弱,而不是完全地丧失。患者临床表现为有自发不规则月经及轻度高血压、低血钾,结合激素测定结果提示肾上腺和性腺保留部分功能。因而患者的基因型与其临床表型是一致的。结论应进行突变P450C17酶的功能学研究来进一步明确结构改变对功能的影响。     

应用微阵列初步分析髓母细胞瘤的基因表达谱

目的　应用微阵列技术研究髓母细胞瘤的分子发病机理。方法　收集新鲜髓母细胞瘤 4例及正常脑组织 1份的组织标本 ,提取总 RNA,逆转录成 32 P标记的 c DNA探针 ,与 Atlas人肿瘤芯片杂交 ,通过放射自显影获得基因谱 ,应用 Atlas Image TM1.0 1a分析。结果　与正常脑组织相比 ,髓母细胞瘤下调基因 6个 ,上调基因 35个 ;逆转录 -聚合酶链反应技术验证结果与芯片检测结果相符。除少数基因外 ,大部分基因的表达趋势与肿瘤生物学特性相符。结论　髓母细胞瘤是与星形细胞起源胶质瘤具有不同分子发病机理的多基因病变 ,不同基因之间可能存在复杂的相互作用和联系 ,值得进一步研究。     

API2-MALT1融合基因变异体在粘膜相关淋巴组织结外边缘区B细胞淋巴瘤中的分布及凋亡的关系

目的探讨API2-MALT1融合基因变异体在粘膜相关淋巴组织结外边缘区B细胞淋巴瘤(extranodal marginal zone B—cell lymphoma of mucosa—associated lymphoid tissue，MALT)中的分布特点及其转录与肿瘤凋亡的关系。方法将逆转录-聚合酶链反应和巢式聚合酶链式反应结合，检测62例不同部位MALT淋巴瘤中API2-MALT1融合基因的多种变异体；通过TdT介导脱氧核苷酸缺口末端标记技术进行肿瘤细胞的原位凋亡检测；通过逆转录-聚合酶链反应和免疫组化染色检测API2的mRNA和蛋白水平。结果62例MALT淋巴瘤中28例检出API2-MALT1融合基因(45．16％)，为变异体A1446-M1123或A1446-M814，但未检出A1446-M541和A1446-M1150。A1446-M1123(18／28)的检出明显多于A1446-M814(10／28)。融和基因转录在甲状腺MALT淋巴瘤中检出最低，在其它部位的分布无差异。在API2-MALT1^ 组(API2-MALT1mRNA表达阳性组)肿瘤凋亡水平明显高于API2-MALT1^-组(API2-MALT1mRNA表达阴性组)，API2的mRNA和蛋白水平低于阴性组。A1446-M1123^ 与A1446-M814^ 病例之间凋亡和API2的变化无差异。结论MALT淋巴瘤中t(11；18)(q21；q21)的发生有部位差异，A1446-M1123可能是中国人MALT淋巴瘤中API2-MALT1融合基因变异体的主要类型。API2-MALT1融合基因转录与MALT淋巴瘤的凋亡水平和API2的变化有关。     

IL-12p40-overexpressing immature dendritic cells induce T cell hyporesponsiveness in vitro but accelerate allograft rejection in vivo: role of NK cell activation and interferon-gamma production

Infusion of genetically modified dendritic cells (DC) expressing immunosuppressive molecules is a potential therapy for organ rejection. IL-12p70, a cytokine produced mainly by DC and macrophages, consists of two subunits, p40 and p35. IL-12p70 is an activator of T cells, while the IL-12p40 subunit serves as a natural antagonist for IL-12p70 action. The primary aim of this study was to evaluate the effect of IL-12p40 gene-modification on both the T-cell stimulatory activity of immature DC (imDC) and their ability to prolong cardiac allograft survival. IL-12p40 gene-modified imDC (DC-p40) exhibited a phenotype characteristic of imDC and displayed impaired T-cell allostimulatory ability in vitro. However, to our surprise, for murine vascularized heterotopic heart transplantation (HHT), administration of donor-derived DC-p40 7 days prior to transplantation did not prolong allograft survival but instead significantly exacerbated cardiac allograft rejection. Further study showed that DC-p40 augmented NK cell activity both in vitro and in vivo and enhanced interferon-gamma (IFN-gamma) production in vivo, which might be due to the increased IL-23 production by DC-p40. Our data suggested that although IL-12p40 gene-modified immature DC can induce T cell hyporesponsiveness in vitro, their ability to activate NK cells and induce IFN-gamma production counterbalances this, exacerbating cardiac allograft rejection. The unexpected effects of DC-p40 limit their value in promoting allograft survival in vivo and likely reflect the complexity of IL-12p40 biology.     

Client feedback on physiotherapy counselling in primary health care

The extent of rehabilitation services is adequate, but little feedback is available on these services and service users. This paper is based on a client feedback survey of physiotherapy services conducted in Spring 2000. The purpose of the paper is to compare patients' perceptions of the physiotherapy instruction they received in a private outpatient facility and in a public physiotherapy facility. The results of the study showed that physiotherapy clients were satisfied with the therapy and instruction they received. They felt they received an adequate amount of instructions and advice. The rehabilitee's strong commitment to care, high motivation and a support network are important to independent rehabilitation in the future. As for the point of contact, the majority of clients in the private physiotherapy facility were gainfully employed while the health centre had almost as many retired clients. Client instruction was not dependent on the point of contact.