A rapid method for determination of growth supporting and toxic hormone levels in plant cell culture

Summary The use of an agar diffusion method for the optimization of the phytohormone concentrations in growth media for plant cell cultures was studied. The method allows a rapid determination of both growth-supporting levels as well as toxic levels of the phytohormones. These levels were determined for aTabernaemontana divaricata cell line and the results could be extrapolated to suspension cultures.     

Characterization of the proteinuria induced by prolonged oral administration of cadmium in female rats

Female Sprague-Dawley rats were given 200 ppm cadmium (Cd) in the drinking water for 11 months. Total proteinuria and the concentrations of Cd in blood, urine, liver and kidney cortex were determined monthly. The proteinuria was characterized by Sephadex G-75 chromatography and by sodium dodecyl sulfate-polyacrylamide gel electrophoresis.

From the 8th month of treatment, the Cd concentration in the kidney cortex levels off at a value of about 250 μg/g wet wt and this phenomenon coincides with the occurrece of proteinuria. The proteinuria was characterized by an increased urinary excretion of high molecular weight (HMW) proteins, particularly γ-globulins. Aminoaciduria also increased which suggests the existence of a slight tubular dysfunction. The renal dysfunction induced by chronic oral administration of Cd seems different from that observed in a previous study in which Cd was administered by the i.p. route (1 mg Cd/kg, 5 times a week for 2 months). For the same level of Cd in the kidney cortex the proteinuria induced by i.p. injection of Cd was usually of mixed type and in some cases of the tubular type. The development of this proteinuria was coincident with the levelling off of Cd concentration in the kidney cortex and in the liver. The saturation of liver by Cd is very likely at the origin of the extensive tubular lesion and of the mixed type proteinuria observed in the i.p. experiment. These results demonstrate the importance of the mode of Cd administration on the nature of the Cd-induced proteinuria in animal. They support also our proposal that both low and HMW proteins should be determined in urine for the early detection of renal damage occurring in man chronically exposed to cadmium.     

Heat stress increases delivery of a unique sub-population of proteins conveyed by fast axonal transport

The effect of heat stress on protein synthesis and fast axonal transport was examined in vitro in bullfrog dorsal root ganglion (DRG) and associated spinal/sciatic nerve. Qualitative and quantitative changes of individual 35S-methionine-labelled proteins were determined following DRG labelling and fast transport in respective nerves via two-dimensional gel electrophoresis/autoradiography. Elevation of temperature from 18 degrees C to 33 degrees C for up to 6 hr resulted in a marked increase in synthesis of five individual DRG species of approximately 74,000 daltons that comigrate with heat shock proteins (HSPs). A quantitative comparison of species within this subset revealed two subgroups differentially affected by stress. The three most basic proteins were induced to approximately 1300% of unstressed controls after 6 hr of stress, while the two most acidic species demonstrated an increase to only 300% of controls over the same period. The relative abundance of 25 additional DRG proteins were uneffected by heat stress. Of 70 35S-labelled fast-transported proteins similarly analyzed, 15, comprising 5 families, were consistently transported at greater than 150% of controls following up to 6 hr of heat stress. Over this period all 15 proteins shared a similar profile of abundance relative to non-induced proteins. Transport was elevated to the greatest extent after 2 hr of stress, declined after 3 hr, and tended to rebound at later times. The remaining 55 fast-transported protein spots analyzed were unaffected. An increased delivery of this unique sub-population of 15 fast-transported proteins suggests a possible involvement in early cellular events that mediate heat stress in the nervous system.     

Use of microcarrier beads to facilitate epithelial outgrowth inside collagen gel

Summary Multicellular aggregates of some epithelial cell types rapidly produce ductlike protuberances when cultured inside a matrix of hydrated collagen gel. However, some epithelioid cells aggregate poorly. These cell types will produce outgrowths rapidly if they are attached to microcarrier beads before embedding them in the collagen matrix. Methods are described for preparing such cultures.     

胃腺癌组织蛋白质双向电泳图谱分析

目的　利用蛋白质组学方法建立分辨率高和重复性好的人胃腺癌组织及其正常胃黏膜组织的双向凝胶电泳图谱 ,并分析其差异表达的蛋白质 ,以期用于早期诊断。方法　采用固相pH梯度(immobilizedpHgradient,IPG)双向凝胶电泳 (two -dimensionalgelelectrophoresis ,2 -DE)分离人胃腺癌组织及正常胃黏膜组织的总蛋白质 ,凝胶经银染显色后 ,ImagingMaster 2D图像分析软件进行比较分析、识别差异表达的蛋白质。结果　 ( 1 )得到了分辨率较高、重复性较好的人胃腺癌组织和正常胃黏膜组织的双向凝胶电泳图谱 ,癌组织和正常胃黏膜组织凝胶的平均蛋白质点数分别为 1 0 4 9± 67和 1 0 97± 73,平均匹配的点数分别为 835± 48和 95 3± 5 6,匹配率分别为 79.6%和 86.7% ;同一组织凝胶在蛋白质点位置上有较好的重复性 ,不同凝胶间蛋白质点在等电聚焦 (isoelectricfocusing ,IEF)方向的偏差是 0 .873±0 .1 2 5mm ,在十二烷基磺酸钠聚丙烯酰胺凝胶电泳 (sodiumdodecylsulfate -polyacrylamidegelelectrophore sis ,SDS -PAGE)方向上的偏差为 1 .0 2 5± 0 .2 1 3mm。 ( 2 )通过比较分析 5例胃腺癌组织及正常胃黏膜组织的双向凝胶电泳图谱 ,得到平均匹配蛋白质点数为 769± 45 ,差异表达蛋白质点数为 81 ,其中 1 7个点仅     

Hip revision surgery in cases with acetabular bone loss: PPR rings, homologous bone and platelet gel

Abstract Hip revision has several objectives: filling the bone defect, restoring the rotational center of the hip, and restoring limb length and hip function. Recently, through tissue engineering, it became possible to consider a fourth objective: to give a graft improved capability to osteointegrate and to restore bone stock as for amount of bone and bone quality (tissue engineering or bioenhancement). Concerning biomechanical and clinical objectives, rings are the most commonly used prosthesis. We used the Partial Pelvic Replacement (PPR) ring and retrospectively analyzed our patients at a mean follow-up of 27.2 months. We found no signs of radiological failure, no radiolucency or osteolysis, nor implant component ruptures. The mean Harris hip score improved significantly from 35.9 preoperatively to 78.1. As for the anatomical objective of hip revision surgery, homologous bone grafts are the most used means to fill a bone defect. We developed a new method to produce platelet gel as a simple and inexpensive way to obtain autologous growth factors, without any discomfort for the patient. We used platelet gel with PPR rings and homologous bone graft; we report our method and describe the first cases treated, with good results.     

硫芥中毒犬外周血IL-2、IL-6含量变化和淋巴细胞DNA损伤

目的 研究硫芥中毒犬外周血IL-2、IL-6变化和淋巴细胞DNA损伤的规律。方法 重庆家犬硫芥中毒(16mg/kg，sc)。中毒后不同时相点取血，分离血清和淋巴细胞。采用放射免疫方法测定血清中IL-2、IL-6含量，以单细胞凝胶电泳技术测淋巴细胞DNA损伤程度。结果 中毒后4h血清IL-2、IL-6含量开始下降，72h达最低值，120h开始恢复。单细胞电泳检测结果提示，硫芥导致明显的淋巴细胞彗星现象。受损细胞率、淋巴细胞DNA片段迁徙度于中毒4h即开始升高，24～72h持续升高。结论 硫芥中毒导致发生早且持续时间较长的犬外周血IL-2、IL-6含量下降和淋巴细胞DNA损伤。     

MGCT与TCT法对新生儿溶血病实验室诊断的分析

目的 :介绍一种新的微柱凝胶抗球蛋白检测技术诊断新生儿溶血病。方法 :采用微柱凝胶技术 (MGCT)和试管抗球蛋白实验 (TCT)同时平行做三项实验进行对比 :①直接抗球蛋白试验 ,②游离IgG抗体试验 ,③放散试验。结果 :MGCT与TCT两种方法检测 98例 (均为Rh阳性 ) ,疑为新生儿溶血病的样本 ,阳性率分别为 6 5 .3% (6 4例 ) ;4 9.9% (4 6例 )。两种方法检测HDN的发生呈显著差异 ,具有统计学意义 (P <0 .0 5 )。结论 :微柱凝胶技术具有敏感性高、操作简便、受人为因素影响少和结果可保存等优点 ,不足之处是价格偏高     

神经系统疾患脑脊液寡克隆IgG区带的检测及其临床意义

聚丙烯酰胺凝胶盘状电泳法检测 CSF 寡克隆 IgG 区带已作为反映中枢神经系统体液免疫异常的定性指标。聚丙烯酰胺凝胶盘状电泳方法作 CSF 寡克隆 IgG 区带分折分辨力高,不需要浓缩 CSF,只要200μg 蛋白的 CSF 就可灵敏地反映出γ-球蛋白质和量的变化.作者应用本方法对124例 CSF 标本检测的结果表明:该法操作简便、灵敏度高,具有一定的临床实用价值。     

Heterogeneity of in vitro growth pattern of megakaryocyte progenitors (CFU-M) in myeloproliferative disorders

In groups of 26 patients with myeloproliferative disorders (MPD), 8 with chronic myelogenous leukaemia (CML); 8 with polycythaemia vera (PV); 10 with essential thrombocythaemia (ET); and 6 patients with reactive thrombocytosis (RT), we studied the growth characteristics of bone marrow CFU-M in agar culture. The bone marrows from all the patients with MPD formed so called endogenous CFU-M colonies, in the absence of PHA-LCM, that increased in a dose-dependent manner with the addition of increasing concentrations of normal human AB-citrated plasma (NH-ABCP), while the bone marrows from all the patients with RT and from healthy controls formed few or no endogenous CFU-M colonies. In MPD, the endogenous CFU-M growth was enhanced by normal T cells in a dose-dependent fashion, and was decreased with the depletion of T cells from the marrow cells. These results suggest that the formation of endogenous CFU-M colonies is caused by hypersensitivity of CFU-M in MPD to NH-ABCP, which may contain a small amount of Meg-CSF, and/or by in vitro T cell stimulation. Among MPD, the endogenous CFU-M growth in ET was significantly lower than that of other MPD patients; however, the total number of ET CFU-M grown in the presence of PHA-LCM was the highest. These data show that the bone marrow CFU-M in MPD are heterogeneous with respect to in vitro growth pattern or sensitivity to exogenous Meg-CSF.