Repetitive Electrical Stimulation of Peripheral Nerve and Spinal Cord Activity

Abstract

Spinal cord neuron and dorsal column fiber responses to electrical stimulation of the sciatic nerve in anesthetized cats were recorded before, during, and after periods of repetitive electrical stimulation of the sciatic nerve through an implantable nerve cuff stimulator device of the type and method used in human patients for pain relief. In previous publications from this laboratory using similar experimental conditions, we reported that repetitive electrical stimulation of the peripheral nerve suppressed all components of the compound action potential of nerves. The present study corrfmns that 5 percent of the spinal cord units studied showed facilitated discharge, 46 percent showed inhibited or depressed discharge, 36 percent underwent no change, and 13 percent showed equivocal responses to repetitive electrical stimulation. Inhibition of dorsal column fiber activity following repetitive electrical stimulation of peripheral nerve is not consistent with the Melzack-Wall gate hypothesis in which suppression of small fiber nociceptive input is mediated by large fiber activity. Our work suggests that the most commonly observed effect 9f electroanalgesia is to cause a more diffuse depreSSIon of nociceptive as well as nonnociceptive spinal cord activity.     

An immuno-precipitation assay for determining specific interactions between antibodies and phage selected from random peptide expression libraries

Libraries of random peptides displayed by bacteriophage can be screened to select phage expressing peptides that specifically bind antibodies, so that the peptide sequence motifs expressed by the phage can help to define the epitopes of the antibodies. It is often desirable to screen antibody-selected phage for binding of the selecting antibody in an immunoassay in order to verify the specificity of the interaction. Enzyme-linked immunosorbent assays (ELISAs) are commonly used for this purpose. However, for many antibodies, the best techniques for measuring specific, high affinity interactions are immuno-precipitation assays. Immuno-precipitation was therefore investigated as a means of measuring interactions between antibodies and phage clones selected from random peptide display libraries. Three mouse monoclonal antibodies specific for glutamic acid decarboxylase were used to select peptides as 9-mers on T7 phage, linear 12-mers on pIII of M13 phage, or constrained 15-mers on pVIII of M13 phage. Following the cloning and sequencing of selected phage, mixtures of antibody and phage were incubated in solution and the immune complexes were precipitated with Protein G bound to Sepharose beads. In order to detect and quantitate the phage that had formed immune complexes and been precipitated, advantage was taken of the biological properties of the phage by inducing infection of Escherichia coli by the precipitated phage. The aim was to quantitate the phage precipitated by determining the number of plaques produced, which would therefore be proportional to the degree of interaction between the phage and the antibody in solution. The results presented here indicate that this method of measuring monoclonal antibody interactions with phage selected for expression of peptides recognised by the monoclonal antibody is highly specific and sensitive.     

小鼠Wnt-3a基因的克隆、测序与真核表达载体的构建

1 材料和方法 1.1 材料 KM小鼠,孕13.5 d,安徽省实验动物中心提供. 大肠杆菌JM109由安徽医科大学分子生物学教研室提供. Trizol试剂盒、pSecTag2/Hygro B质粒(Invitrogen). 质粒小量提取试剂盒、 RT-PCR试剂盒(Promega);Ex Taq DNA聚合酶、T4 DNA连接酶,DNA Marker, pMD18-T Vector,限制性内切酶(TaKaRa).     

Vacuum-assisted stereotactic breast biopsy: histologic underestimation of malignant lesions

HYPOTHESIS: The histopathologic correlation between stereotactic core needle biopsy and subsequent surgical excision of mammographically detected nonpalpable breast abnormalities is improved with a larger-core (11-gauge) device. DESIGN: Retrospective medical record and histopathologic review. SETTING: University-based academic practice setting. PATIENTS: Two hundred one patients who underwent surgical excision of mammographic abnormalities that had undergone biopsy with an 11-gauge vacuum-assisted stereotactic core biopsy device. MAIN OUTCOME MEASURE: Correlation between stereotactic biopsy histologic results and the histologic results of subsequent surgical specimens. RESULTS: Results of stereotactic biopsy performed on 851 patients revealed atypical hyperplasia in 46 lesions, ductal carcinoma in situ (DCIS) in 89 lesions, and invasive cancer in 73 mammographic abnormalities. Subsequent surgical excision of the 46 atypical lesions revealed 2 cases of DCIS (4.3%) and 4 cases of invasive carcinoma (8.7%). Lesions diagnosed as DCIS on stereotactic biopsy proved to be invasive carcinoma in 10 (11.2%) of 89 patients on subsequent excision. Stereotactic biopsy completely removed 21 (23.6%) of 89 DCIS lesions and 20 (27.4%) of 73 invasive carcinomas. CONCLUSIONS: In summary, 11-gauge vacuum-assisted core breast biopsy accurately predicts the degree of disease in the majority of malignant lesions; however, understaging still occurs in 11% to 13% of lesions showing atypical hyperplasia or DCIS.     

Low temperature incorporation of bovine serum albumin into a bead formed macroporous hydrophilic polymer matrix with potential for sustained release.

A process of freeze-thaw polymerization involving the low temperature photopolymerization of a mixed solution of monomers and bovine serum albumin around frozen ice crystals has been used to generate a bead formed macroporous hydrophilic matrix with potential for sustained release. Beads over the size range 100-3000 microns were fabricated with surface and internal pores of between 0.7-2.6 microns whose diameter could be controlled by manipulation of the monomers to solvent ratio. Increasing both the proportion of monomers in the monomer solution and the percentage of BSA incorporated reduced the EWC of beads. The BSA release profile was characterized by an initial burst followed by a lower but sustained release lasting up to 1 month. The total cumulative release of BSA and the proportion of the incorporated BSA load subsequently released were both reduced in physiological saline compared with distilled water but enhanced by freeze drying, mild agitation and incubation at 37 degrees C.