瘢痕疙瘩家系Fas基因的突变：2个家系10份标本分析

目的:观察瘢痕疙瘩家系样本中Fas基因有无突变,探讨Fas基因突变在瘢痕疙瘩形成中的意义。方法:实验于2005-01/05在上海基康公司完成。①标本来自南方医科大学南方医院整形外科2005年收集的A和B两个瘢痕疙瘩家系,所有参与观察的家系成员均签署知情同意书。②采用聚合酶链反应及基因测序技术,分别以A家系两例患者的瘢痕疙瘩组织为观察对象,以其周围正常皮肤及外周静脉血作为自身对照;其配偶的外周静脉血作为正常对照。并以B家系中两例患者的外周静脉血作为不同家系间的对照。共取10份样本,4份组织样本,6份静脉血标本。检测10份样本中Fas基因外显子1～9的基因序列。结果:①基因测序发现所检测的10个瘢痕疙瘩家系标本Fas基因的1～8外显子均未发现突变。②2份瘢痕疙瘩组织标本在第9外显子编码区的11bp,53bp两个位点上存在单个碱基的基因突变或多态性改变。结论:瘢痕疙瘩Fas基因外显子9区段的基因结构异常极有可能造成Fas蛋白的功能改变,从而导致身体局部瘢痕疙瘩的形成。     

H-2-restricted cytotoxic effectors generated in vitro by the addition of trinitrophenyl-conjugated soluble proteins

Murine spleen cells from normal donors were cultured in vitro with trinitrobenzene sulfonate (TNBS)-conjugated soluble proteins, i.e., bovine gamma globulin (TNP-BGG) or bovine serum albumin (TNP-BSA). Addition of 100 μg of any of these TNP-proteins to the spleen cell cultures led to the generation of cytotoxic T-cell effectors which were H-2-restricted and TNP- specific. The lytic potential of such effectors was comparable to that generated by sensitization with TNBS-modified syngeneic cells, and was restricted to haplotypes shared at the K or K plus I-A, or the D regions of the H-2 complex. Greater effecter cell activity was generated by addition of TNP-BGG against TNBS-modified targets which shared K plus I-A than against modified targets which shared the D region with the responding cells, which suggests that the same immune response genes are involved when the response is generated by the addition of TNP-conjugated soluble proteins or of TNBS- modified cells. H-2-restricted, TNP-specific effecter cells were generated by culturing mouse spleen cells with syngeneic cells which had been preincubated with TNP- BGG or TNP-BSA for 1.5 h. The addition of unconjugated soluble proteins to the cultures did not result in cytotoxic effectors detectable on H-2-matched targets, whether the targets were prepared by modification with TNBS, or by incubation with either the unconjugated or TNP-conjugated proteins. Depletion of phagocytic cells in the tumor preparation by Sephadex G-10 column fractionation before incubation with TNP-BSA had no effect on their lysis by the relevant effector cells. Immunofluorescent staining of tumor target cells with anti-TNP antibodies indicated that TNP could be detected on the tumor cells within 10 rain of incubation with TNP-BSA. The cytotoxic response generated by addition of the TNP-proteins to spleen cell cultures was found to be T-cell dependent at the effector phase, as shown by the sensitivity of the lytic phase to absorbed RAMB and complement. Furthermore, the response did not appear to be attributable to antibody-dependent cellular cytotoxicity. Three mechanisms were considered which could account for the generation of H-2-restricted, TNP-specific, cytotoxic T-cell effectors by the addition of soluble TNP-proteins. These include covalent linkage of activated TNP groups from the soluble proteins to cell surface components, macrophage processing of the soluble conjugates and presentation to the responding lymphocytes in association with H-2-coded self structures, or hydrophobic interaction of the TNP-proteins to cell surfaces. Results obtained from sodium dodecyl sulfate gel patterns indicating that cell-bound TNP was still linked to BSA, and the observation that phagocytic-depleted cells could interact with the soluble TNP-proteins and function as H-2-restricted targets, appear not to favor the first two proposed mechanisms.     

跟骨定量超声骨质测量参数与骨密度及骨组织形态计量学指标的关系

目的:分析跟骨定量超声骨质测量中各参数与骨密度及形态计量学指标的相关性。方法:选择2004-01/2005-12广州市第六人民医院和中山大学三院骨科小腿以上截肢患者38例,将其跟骨定量超声测定的超声振幅衰减平均值与健康青年人骨峰值进行比较,>-2.5 SD者为骨量正常组(12例),<-2.5 SD者为骨质疏松组(26例)。分别进行跟骨定量超声、双能X线骨密度测量仪及骨形态计量学测量,应用直线相关分析法分析跟骨定量超声测定中各参数与骨密度及骨组织形态计量学各指标的相关性。结果:38例全部进入结果分析。①骨质疏松组跟骨超声振幅衰减平均值和骨硬度指数值均小于骨量正常组(P<0.01)。②骨量正常组跟骨骨密度值显著高于骨质疏松组[(352±16),(233±14)mg/cm2,P<0.01]。③骨量正常组跟骨平均骨小梁间距或弥散度低于骨质疏松组而松质骨体积高于骨质疏松组(P<0.05)。④超声振幅衰减平均值和骨硬度指数与骨密度呈直线正相关(r=0.814,0.326,P<0.01,0.05)。⑤超声传播速度与骨小梁游离末端、平均骨小梁间距呈直线负相关(r=-0.688,-0.712,P<0.01),与小梁间连点数、松质骨体积呈直线正相关(r=0.672,0.794,P<0.01);骨硬度指数与平均骨小梁间距呈直线负相关(r=-0.358,P<0.05),与松质骨体积呈直线正相关(r=0.513,P<0.01)。结论:跟骨定量超声测量中,超声振幅衰减平均值能较好地反映骨的密度,超声传播速度能较好地反映骨的质量,而骨硬度指数能较综合地反映骨强度的改变。     

The Diurnal Variation of the Serum Iron Concentration

Observations made in an investigation of the diurnal variation of the serum iron concentration suggested that the diurnal variations of serum iron can be explained as a phenomenon secondary to the diurnal variation of the hemoglobin metabolism. This hypothesis is supported by parallel changes in the bilirubin and serum iron values.     

Erg?nzung und Unterstützung zu konventionellen Tumortherapien Misteltherapie

Die anthroposophische Misteltherapie kann etablierte onkologische Standardtherapien im Sinne einer umfassenderen und ganzheitlichen Versorgung erg?nzen.     

In vitro characterization of the human recombinant soluble granulocyte- macrophage colony-stimulating factor receptor

We have cloned, expressed, and partially purified a naturally occurring, truncated, soluble form of the human granulocyte-macrophage colony-stimulating factor (GM-CSF) receptor alpha subunit to investigate its biochemical and biologic properties. The soluble receptor species lacks the transmembrane and cytoplasmic domains that are presumably removed from the intact receptor cDNA by a mechanism of alternative splicing. The resulting soluble 55- to 60-kD glycosylated receptor species binds GM-CSF with a dissociation constant (kd) of 3.8 nmol/L. The soluble GM-CSF receptor successfully competes for GM-CSF binding not only with the transmembrane-anchored GM-CSF receptor alpha subunit but also with the native oligomeric high-affinity receptor complex. In addition, in human bone marrow colony-forming assays, the soluble GM-CSF receptor species can antagonize the activity of GM-CSF. Our data suggest that the soluble GM-CSF receptor may be capable of acting in vivo as a modulator of the biologic activity of GM-CSF.     

Corticotropin-releasing factor concentrations exhibit an apparent diurnal rhythm in hypothalamic and extrahypothalamic brain regions: differential sensitivity to corticosterone

Regional brain concentrations of corticotropin-releasing factor (CRF) exhibited marked differences in a number of regions depending upon the time of day rats were sacrificed. When compared with animals killed at 09.00 h, CRF concentrations at 15.30 h were elevated in the median eminence, hypothalamus minus median eminence, preoptic area/suprachiasmatic nucleus, bed nucleus of the stria terminalis, septum, frontal/parietal cortex, cerebellum, cingulate cortex, locus ceruleus and the nucleus of the solitary tract. Animals that received 7 days of continuous corticosterone supplementation via osmotic minipump either did not exhibit an afternoon rise in CRF concentrations (median eminence, cingulate cortex and locus ceruleus) or exhibited significantly attenuated afternoon increases in CRF concentrations (hypothalamus minus median eminence). In rats killed at 09.00 h, corticosterone reduced CRF concentrations in the median eminence. Chronic corticosterone exposure did not affect CRF concentrations in any of the extrahypothalamic brain regions studied in rats killed at 09.00 h. These results suggest that a number of CRF-containing neurons in both hypothalamic and extrahypothalamic brain regions presumably undergo diurnal changes in their activity as evidenced by changes in CRF concentrations. Furthermore, chronic corticosterone supplementation can alter these changes, particularly those in the endocrine hypothalamus.     

A longitudinal immunologic evaluation of hemophiliac patients

Over an average span of one year, we performed a prospective clinical and immunologic evaluation of 30 patients with hemophilia. No patient developed life-threatening opportunistic infection or malignancy; however, the immunologic abnormalities and lymphadenopathy initially present in nine patients (lymphadenopathy group) persisted. In addition, five patients, representing 24% of the initial group without lymphadenopathy, developed generalized lymphadenopathy (converter group). One episode of idiopathic thrombocytopenia (ITP) and one episode of staphylococcal sepsis occurred in this "converter" group; one episode of ITP also occurred in the lymphadenopathy group. Sixteen patients remained asymptomatic. At the time of the follow-up evaluation, those differences in mononuclear cell (MNC) percentages and numbers noted initially among the three hemophiliac groups were no longer present. Natural killer cell function alone or in the presence of biologic response modifiers was not different among hemophiliac and control groups. Before developing lymphadenopathy, the converter group of patients had significantly better lymphocyte mitogenic function than did the other two groups of patients with hemophilia. However, lymphocyte mitogenic responses of all groups of patients with hemophilia significantly deteriorated over the course of the study. The abnormal mitogenic responses noted in these patients was explained in part by higher levels of spontaneous suppressor cell activity in mononuclear cell preparations from patients with hemophilia. We conclude that long-term immunologic studies of this patient population requires both quantitative and qualitative evaluations. Our data show that patients with hemophilia have progressive dysfunction of cell- mediated immunity.     

Marked reduction in gonadal steroid hormone levels in rats treated neonatally with monosodium L-glutamate: further evidence for disruption of hypothalamic-pituitary-gonadal axis regulation

Serum levels of luteinizing hormone (LH), follicle stimulating hormone (FSH), prolactin, estradiol-17 beta and testosterone were determined in adult rats that were treated in the neonatal period with monosodium L-glutamate (MSG) which has previously been shown to reliably produce destruction of arcuate nucleus perikarya. MSG-treated males had significantly smaller accessory sexual organs (seminal vesicles and ventral prostate) and tests and had significantly lower serum concentrations of FSH and testosterone than sex-matched controls. MSG-treated females had significantly lower serum concentrations of LH, FSH and estradiol-17 beta. Prolactin levels of MSG-treated rats were no different than sex-matched controls. This marked reduction in gonadal steroid levels (decreases 68%) and inappropriately low gonadotropin levels further characterizes the deficit of feedback regulation in the hypothalamic-pituitary-gonadal axis in MSG-treated rats.