ABSTRACT
Objective To prepare monoclonal antibody(McAb) against human tissue kallikrein (HK) and develop an ELISA kit allows for the in vitro quantitative determination of human tissue kallikrein in urine. Methods To generate a monoclonal antibody specific for TK, the synthetic TK peptide consisting of 12 amine acids(12P), was fused to keyhole limpet hemocyanin(KLH) and used for immunization. Using hybridoma screening, monoclonal secreting cell lines were identified and used to generate ascites in BALB/c mouse. Antibody was purified by affinity column chromatography. 12% SDS-PAGE and Western blot were used to visualize the purified antibody. This kit employs indirect competitive ELISA technique and BiotinAvidin System. 12P was fused to bovine serum albumin(BSA) and has been pre-coated onto a microplate at first. Standards and samples were added to the appropriate microplate wells with a biotin-conjugated McAb croplate well. A TMB substrate solution is added to each well. The enzyme-substrate reaction is terminating by the addition of a sulphuric acid solution and the color change is measured spectrotometrically at a wavelength of 450 nm. The concentration of tissue kallikrein in the samples is then determined by comparing the O.D. of the samples to the standard curve. Results 8 hybridoma cell lines secreting mAbs special to HK,SDS-PAGE and Western blot demonstrated successful preparing and purification of McAb( 100% ). The linearity of this ELISA kit is demonstrated(r =0. 990). The range of detection of the assay is 0.008 μg/ml to 0. 5 μg/ml. The assay remained stable, with no change in the values measured, over five cycles of freezing and thawing. Conclusion 8 McAbs against HK have been prepared successfully and possess high titer and specificity. The development of an ELISA kit for detecting HK can meet the needs of detection of HK in urine samples.
ABSTRACT
@#ObjectiveTo evaluate the assistant effct of electroacupuncture on acute incomplete spinal cord injury treated with routine therapy.Methods9 cases who accepted routine therapy, including operation and medicine, were in control group, while other 7 cases who accepted electroacupuncture other than routine therapy were in treating group.ResultsThere were 2 cases were effective, 4 cases were improved and 3 cases were not improved in control group, while 6 cases were clinical recovery, 1 case was effective in treating group.ConclusionElectroacupuncture can improve the effect of routine therapy on acute incomplete spinal cord injury.
ABSTRACT
@# 目的观察神经外科与针刺联合治疗急性脊髓不完全损伤的疗效。方法神经外科治疗组根据指征确定是否进行手术,药物治疗均给予激素、脱水、抗去甲肾上腺素类药物、神经营养药物,高压氧。神经外科与针刺联合治疗组除上述神经外科治疗外,采用针刺治疗,针刺处方:电针刺激夹脊穴,根据不同辨证配合远端取穴。结果神经外科治疗组9例,持续治疗 3—6个月,出院后随访6个月,6例好转,肌力有所改善,最好者可达到肌力IV—IV+级,3例无效,甚至退步,瘫痪进一步加重。神经外科与针刺联合治疗组病人5例,经针刺及电针治疗10—80天时间后,4例临床治愈,1例经针刺治疗好转后,因转入骨科手术治疗腰椎压缩性骨折时脱离针刺治疗。结论神经外科与针刺联合治疗急性脊髓损伤效果好,并能抑制脊髓损伤的进一步恶化。