A model of corrective gene transfer in X-linked ichthyosis

Single gene recessive genetic skin disorders offer attractive prototypes for the development of therapeutic cutaneous gene delivery. We have utilized X-linked ichthyosis (XLI), characterized by loss of function of the steroid sulfatase arylsulfatase C (STS), to develop a model of corrective gene delivery to human skin in vivo. A new retroviral expression vector was produced and utilized to effect STS gene transfer to primary keratinocytes from XLI patients. Transduction was associated with restoration of full-length STS protein expression as well as steroid sulfatase enzymatic activity in proportion to the number of proviral integrations in XLI cells. Transduced and uncorrected XLI keratinocytes, along with normal controls, were then grafted onto immunodeficient mice to regenerate full thickness human epidermis. Unmodified XLI keratinocytes regenerated a hyperkeratotic epidermis lacking STS expression with defective skin barrier function, effectively recapitulating the human disease in vivo. Transduced XLI keratinocytes from the same patients, however, regenerated epidermis histologically indistinguishable from that formed by keratinocytes from patients with normal skin. Transduced XLI epidermis demonstrated STS expression in vivo by immunostaining as well as a normalization of histologic appearance at 5 weeks post-grafting. In addition, transduced XLI epidermis demonstrated a return of barrier function parameters to normal. These findings demonstrate corrective gene delivery in human XLI patient skin tissue at both molecular and functional levels and provide a model of human cutaneous gene therapy.      

新复合纤维蛋白胶可注射性磷酸钙人工骨的理学特性

目的:检测纤维蛋白胶复合β-磷酸三钙/磷酸二氢钙复合人工骨材料的物理学性能,评价纤维蛋白胶对β-磷酸三钙/磷酸二氢钙骨水泥性能的影响,以及其作为注射型复合人工骨用于修复骨缺损的可行性。方法:实验于2006-12/2007-06在南方医科大学珠江医院中心实验室和华南理工大学生物材料实验室完成。①材料:β-磷酸三钙由上海瑞邦生物材料有限公司提供,磷酸二氢钙为东泰化工赠,纤维蛋白胶购自广州倍绣生物技术有限公司。②复合材料制备:将β-磷酸三钙/磷酸二氢钙骨水泥按3∶1的比例充分混合后,与纤维蛋白胶按凝固后的体积2∶1体积比混合,制成复合人工骨材料。③观察指标:测定复合材料的凝固时间,抗压强度,抗稀散性能,并用扫描电镜观察其煅烧前后的显微结构特征,以未加纤维蛋白胶的磷酸钙水泥为对照(CPC组)。结果:复合人工骨材料的平均初凝时间长于CPC组(P<0.004),终凝时间在初凝时间后2～4 min;复合材料的抗压强度为(14.72±1.81)MPa。复合材料较CPC组有良好的抗稀散性能,扫描电镜发现,纤维蛋白胶贯穿于磷酸钙水泥晶体间,并将磷酸钙水泥晶体紧密连接。煅烧后复合材料的孔径有增大,空隙率为57.28%,并且微孔之间有空隙互相贯通。结论:该骨水泥复合材料凝固时间符合临床操作的需要;抗压强度达到松质骨强度的要求;煅烧后磷酸钙水泥的空隙率明显提高,有利于材料的降解。     

薄层扫描法测定黄芪生脉颗粒中黄芪甲甙含量

目的：制订黄芪生脉颗粒中黄芪甲甙含量测定方法。方法：双波长薄层扫描法，经乙酰洗涤、正丁醇提取和Ｄ１０１大乳吸附树脂柱层析法制备样品，以氯仿－甲醇－水（１３：７：２）下层液为展开剂，检测波长为５１０ｎｍ，参比波长为７００ｎｍ。结果：加标回收率平均为９８．７％（ＲＳＤ＝２．０％，ｎ＝６），标准曲线ｒ＝０．９９６６，重复性ＲＳＤ＝１．４％（ｎ＝５），精密度ＲＳＤ＝２．０％（ｎ＝６）。结论：方法稳定、可靠     

新型缩氨基硫脲衍生物——氨基巯三唑Schiff碱的合成与抗菌活性

设计合成了新型缩氨基硫脲衍生物4-氨基-3-(呋喃-2)-5-巯基-1,2,4-三唑及其十种不同醛的Schiff碱衍生物,并进行了抑菌实验。结果表明:Schiff碱化合物结构中的甲亚胺基是该类化合物的活性功能基。苯环上更换不同的取代基对其活性有一定的影响,其中化合物4-(5-硝基亚糠基氨基)-3-(呋喃-2)-5-巯基-1,2,4-三唑(Ik)抗菌活性最高且抗菌谱最广;化合物4-亚水杨基氨基-3-(呋喃-2)-5-巯基-1,2,4-三唑(Ib)具有较强的抑霉活性。     

Rotator cuff lesions of the shoulder: evaluation by direct sagittal CT arthrography

Direct sagittal computed tomographic scanning (DSCT) of the shoulder was performed in 42 symptomatic patients, six healthy volunteers, and two cadaver shoulders. Axial CT scanning and double-contrast arthrography with plain radiographs were performed in 41 patients for comparison. DSCT enabled correct identification of 27 of 29 lesions in 24 patients. Seventeen patients had normal shoulders. Axial CT scanning and DSCT together enabled correct identification of all lesions and were markedly superior to plain-film arthrography. DSCT enabled diagnosis of all cases of complete rotator cuff tear plus three cases of incomplete tear and three of rotator cuff atrophy not identified by the other techniques. Axial CT scanning was better than DSCT for diagnosis of Bankart lesions.     

Negative charge distribution and density on the surface of oxygenated normal and sickle red cells

Negative charges on the external surface of red cells were visualized by colloidal iron hydroxide labelling of 50% of the membrane area after osmotic hemolysis and glutaraldehyde fixation. Counts were made over randomly selected areas on electron micrographs at 350,000 x magnification. Statistical analyses showed that at the 95% level of confidence there was no significant difference between oxygenated normal (AA) and sickle (SS) cells in either the distribution or the density of negative charges.     

Chlorpropamide-induced pure white cell aplasia

We investigated the mechanism for isolated agranulocytosis and marrow pure white cell aplasia in an elderly man receiving 0.5 to 1.0 g per day of chlorpropamide (Chl) without other toxic drug exposure or overt systemic illness. Patient marrow revealed an absence of recognizable granulocytic precursors; megakaryocytes and erythroid precursors were normal. The WBC count was 1800/mm3 on admission with only 2% neutrophils; the absolute neutrophil count first exceeded 500/mm3 on the 17th day following cessation of Chl. A serum Chl level on admission was 100 micrograms/mL (acute phase, AP); no Chl was detected in serum (convalescent phase, CP) assessed on the 22nd hospital day. Antineutrophil antibodies were not detected, and T cell depletion failed to augment patient in vitro granulopoiesis. Patient AP serum produced potent complement-mediated inhibition (87% +/- 7%) of autologous granulocyte progenitors (CFU-GM) with minimal inhibition of erythroid (11% +/- 5%) or multipotent (5% +/- 4%) progenitor cells. Selective inhibition by patient AP serum of CFU-GM (74% +/- 11%) was also seen against two allogeneic marrows. Patient CP serum no longer inhibited (6% +/- 4%) autologous CFU-GM. Addition of Chl (5 to 120 micrograms/mL) to CP serum but not to control serum resulted in potent drug concentration-dependent complement-mediated inhibition of autologous and allogeneic CFU-GM. Inhibition of CFU-GM in the presence of Chl was no longer demonstrable following immunoabsorbent removal of IgG from patient serum. Patient serum in the presence of Chl had limited activity against morphologically recognizable marrow granulocytic precursors in a microimmunofluorescence assay. These results are most consistent with the development of Chl-dependent, selective antibody-mediated immune inhibition of granulopoiesis.     

Lumbar iohexol myelography and diagnostic lumbar puncture. Headache and associated side effects in relation to neurological signs and diagnosis, previous mental symptoms and pain history

Various possible risk factors for postlumbar puncture (and postiohexol-myelographic) headache and associated side effects were analysed. Headache and nausea occurred significantly more often in patients without clinical findings than in those with findings. We found significantly different incidences of severe headache and nausea between diagnostic subgroups after a lumbar puncture. The greatest headache incidence was found in patients without a definite neurological diagnosis, while nausea occurred most frequently in patients with various painful disorders. Following iohexol myelography, nausea occurred most often in patients who had a history of previous mental symptoms and in patients with a history of previous headache disorders. Mental symptoms were more frequently reported in patients who also had experienced mental symptoms previously. The relationship between side effects and negative clinical findings was stronger than the relationship between side effects and previous mental symptoms.