高压液相色谱法纯化抗人大肠部单克隆抗体

采用Ｂｉｏ－Ｒａｄ公司的ＭＡＰＳ１００单克隆抗体纯化系统，利用羟基磷灰石层新 柱对鼠柱对复腹水中抗人大肠癌单克隆抗体进行分离纯化。结果：还原性ＳＤＳ－ＰＡＧＥ显示所获抗体为单一组分，ＥＬＩＳＡ证实抗体活性满意，所获抗体收率为２３％－５２％，重复进样收率稳定（３８．６３±２．２６）％，ｎ＝３。     

McAb—BA—ELISA及McAb—ELISA检测青霉素特异IgE及其评价

用单克隆抗体及生物素与亲和素，建立了ＭｃＡｂ－ＢＡ－ＥＬＩＳＡ（单克隆抗体－生物素亲和素－酶联免疫吸附试验）和ＭｃＡｂ－ＥＬＩＳＡ（单克隆抗体酶联免吸附试验），用于检测Ⅰ型青霉素过敏患者中，ＢＰＯ－Ｓｐｅｃ－ＩｇＥ阳性率为４５％，ＢＰＯ－Ｓｐｅｃ－ＩｇＥ阴性者中，用ＭｃＡｂ－ＥＬＩＳＡ检测ＢＰＥ－Ｓｐｅｃ－ＩｇＥ与６－ＡＰＡ－Ｓｐｅｃ ＩｇＥ，阳性率分别为１２．５％及３５．０％，总阳性率为９２．…     

SDS—PAGE法分离纯化牛角膜上皮特异性膜结合蛋白质55.3kD

用ＳＤＳ－ＰＡＧＥ法离纯化了牛角膜上皮特异性膜结合蛋白质５５．３ｋＤ，结果显示，ＳＤＳ－ＰＡＧＥ可用于从少量样品中提高纯度的蛋白质，且有效好的收率。     

SDS－PAGE法分离纯化牛角膜上皮特异性膜结合蛋白质55．3kD

用ＳＤＳ－ＰＡＧＥ法离纯化了牛角膜上皮特异性膜结合蛋白质５５．３ｋＤ，结果显示：ＳＤＳＰＡＧＥ可用于从少量样品中提取高纯度的蛋白质，且有较好的收率。     

胆固醇酯转运蛋白的提纯和鉴定

目的：分离纯化人血清胆固醇酯转运蛋白（ＣＥＴＰ），为制备ＣＥＴＰ单克隆抗体和ＣＥＴＰ浓度测定提供物质基础。方法：制备无脂血清，用Ｂｕｔｙｌ－Ｓｅｐｈａｒｏｓｅ４ＦＦ、ＣＭ－ＳｅｐｈａｒｏｓｅＦＦ和ＳｅｐｈａｃｒｙｌＳ－２００三步柱层析分离、纯化ＣＥＴＰ。结果：纯化的ＣＥＴＰ经ＳＤＳ－ＰＡＧＥ与ＣＥＴＰ特异性单克隆抗体免疫反应和活性测定等鉴定证实为纯化ＣＥＴＰ。结论：经三步柱层析获得了纯化ＣＥＴＰ，得率约２５％，活性３０ｎｍｏｌ／（μｇ·ｈ）。     

重组汉坦病毒核壳蛋白的制备及其单克隆抗体的研制

将ＰＣＲ扩增的ＨＴＶ７６－１１８ＲＮＡ小片段的ｃＤＮＡ克隆插入到载体ｐＤＳ５６／ＲＢＳⅡ－（Ｏ）－６Ｈｉｓ中,通过ＩＰＴＧ诱导表达以及镍螯合层析纯化重组ＮＰ,经过ＳＤＳ－ＰＡＧＥ、免疫转印和ＥＬＩＳＡ方法分析其蛋白特性,证实该重组ＮＰ与毒粒ＮＰ相同,均为４９．６ｋｕ,能与肾综合征出血热（ＨＦＲＳ）患者血清产生特异性反应。以此重组ＮＰ作为抗原制备抗ＮＰ单克隆抗体,获得两株持续稳定分泌单克隆抗体的杂交瘤细胞株。用抗ＮＰＭｃＡｂ建立检测ＩｇＭ抗体的ＥＬＩＳＡ捕获法。证实纯化重组ＮＰ作为抗原,避免了组织培养病毒的感染性和非特异性以及难以标准化的缺点。所建立的ＥＬＩＳＡ捕获法检测ＨＦＲＳ患者血清中ＩｇＭ抗体的方法特异性强、敏感性高,可作为ＨＦＲＳ的早期诊断试剂     

PVC薄膜SPA—ELISA诊断包虫病的应用研究

本文以纯化的人肝棘球蚴囊液致敏ＰＶＣ薄膜板，液体的ＨＲＰ－ＳＰＡ为第二抗体，敏感无毒的ＴＭＢ做底物，建立了ＰＶＣ薄膜ＳＰＡ－ＥＬＩＳＡ法，检测７２例手术证实的包虫病患者血清抗体，阳性符合率为９１．１７％（６６／７２），４５例临床确诊的非包虫病人２例呈阳性，６０例健康人均阴性，假阳性率为１．８３％（２／１０５），结果表明该法即保持了经典ＥＬＩＳＡ敏感性高，特异性强的优点，而且操作简单，快速，无毒，省     

阳离子交换色谱一步法制备级纯化单克隆抗体

应用快速蛋白液相色谱系统（ＦＰＬＣ）建立一种一步法制备级纯化单隆抗体方法。方法：将ＭｃＡｂ腹水用ｐＨ６．０，０．０１ｍｏｌ／Ｌ ＰＢ透析或稀释８倍后直接上ＳＰ－４０ＨＲ阳离子交换色谱柱，用ＰＢ－ＮａＣｌ二元线性离子梯度洗脱，即得到纯化的ＭｃＡｂ。结果：每次上样量７０～８０ｍｌ腹水，可获得纯化ＭｃＡｂ５００～５８０ｍｇ，得率７～７．５ｍｇ／ｍｌ腹水，回收率为７０％～７５％，ＳＤＳ－ＰＡＧＥ检测纯度为     

CyP A三类免疫球蛋白自身抗体测定方法的建立与临床应用

目的：利用基因重组技术、大肠杆菌表达人环孢菌素Ａ结合蛋白（ｒｈＣｙＰＡ），建立测定抗ＣｙＰＡ自身抗体的酶联免疫吸附试验（ＥＬＩＳＡ），探讨抗ＣｙＰＡ抗体与临床疾病的关系。方法：重组菌可溶性蛋白经饱和硫酸铵盐析和ＤＥＡＥ－ＳｅｐｈａｒｏｓｅＣＬ－６Ｂ柱层析，获取ｒｈＣｙＰＡ蛋白，用该纯化物作为抗原，方阵滴定，建立ＥＬＩＳＡ技术，对各类自身免疫病进行评估测定。结果：ｒｈＣｙＰＡ经十二烷基硫酸钠－聚丙烯酰胺凝胶电泳（ＳＤＳ－ＰＡＧＥ）分析，分子量为１８０００。在系统性红斑狼疮（ＳＬＥ）患者中，发现有４０．３％的患者检出有抗ＣｙＰＡ自身抗体（５８／１４４），三类免疫球蛋白（Ｉｇ）中，又以ＩｇＧ类抗体检出率最为突出，为２８％（４０／１４４）；而其它自身免疫病患者抗ＣｙＰＡ抗体总体检出率为０％～１８．１％。结论：建立的检测抗ＣｙＰＡ抗体的ＥＬＩＳＡ法，敏感、特异、可半定量测定。抗ＣｙＰＡ抗体的存在似与ＳＬＥ的发病有关。     

Tamm－Horsfall糖蛋白ABC－ELISA测定法的建立

人尿Ｔａｍｍ－Ｈｏｒｓｆａｌｌ糖蛋白（ＴＨＰ）经盐析、ＳｅｐｈａｄｅｘＧ－１００层析获得ＳＤＳ－ＰＡＧＥ均一的蛋白质纯品。后者与福氏完全佐剂充分混合免疫家兔得到抗ＴＨＰ抗体。将纯化了的该抗体与生物素连结，建立了生物素－抗生素系统的ＥＬＩＳＡ（ＡＢＣ－ＥＬＩＳＡ）测定法。该法最低检测限为２ｎｇ，回收率９７％，变异系数４．７的。测定５１名正常人血清中ＴＨＰ水平为２２６．９４±１４５．９０ｎｇ／ｍｌ。并测定了５０名各类肾脏病人血中的ＴＨＰ值。     

Value of D-Dimers in patients with acute aortic dissection

Objective: To evaluatel the value of D-dimers in patients with acute aortic dissection (AAD). Methods: This study consisted of 16 patients with AAD and 27 non-AAD patients. Serum D-dimets were measured by Sta-Liatest D-DI immunoturbidimetric assay. Results: D-dimer level was higher (P ＜ 0.001) in patients with AAD(7.91 ± 5.52 μg/ml) than that in non- AAD group(1.57±1.24 μg/ml). D-dimer was positive (＞0.4 μg/ml) in all patients with AAD and in 10 control group patients (37%). Among patients with acute AAD, D-dimers tended to be higher in Stanford A than in Stanford B (8.67 ± 4.31 μg/ml vs. 3.24±1.27 μg/ml, P ＜0.01). D-dimer values tended to be higher in more extended disease(3.84 ± 1.65 μg/ml, 8.57 ± 3.58 μg/ml and 11.87 ± 5.69 μg/ml in thoracic aorta, thoracic and abdominal aorta, thoracic and abdominal aorta and iliacal arteries, respectively, P ＜ 0.05 for both 8.57 ± 3.58 and 11.87 ± 5.69 vs. 3.84 ± 1.65 ). Including the control group into the analysis, we found a sensitivity of 100%, a negative predictive value of 100%, and a specificity of 66% and a positive predictive value of 64% for D-dimer in diagnosis of AAD in our patients with suspected AAD. Conclusion: D-dimer was elevated in patients with AAD. A negative D-dimer test result could be useful in excluding AAD.     

Research of combined liver-kidney transplantation model in rats

Objective： To set up a simple and reliable rat model of combined liver-kidney transplantation. Methods： SD rats served as both donors and recipients. 4℃ sodium lactate Ringer＇s was infused from portal veins to donated livers,and from abdominal aorta to donated kidneys, respectively. Anastomosis of the portal vein and the inferior vena cava （IVC） inferior to the right kidney between the graft and the recipient was performed by a double cuff method, then the superior hepatic vena cava with suture. A patch of donated renal artery was anastomosed to the recipient abdominal aorta. The urethra and bile duct were reconstructed with a simple inside bracket. Results： Among 65 cases of combined liver-kidney transplantation, the success rate in the late 40 cases was 77.5%. The function of the grafted liver and kidney remained normal. Conclusion： This rat model of combined liver-kidney transplantation can be established in common laboratory conditions with high success rate and meet the needs of renal transplantation experiment.     

APPLICATION OF LORNOXICAM TO PATIENT-CONTROLLED ANALGESIA IN PATIENTS UNDERGOING ABDOMINAL SURGERIES

FOR anesthesiologis s ,treatingpostoperativepainhas alwaysbeen a problem.Althoughopioidshave been provedtobe effective,theirsideeffectscouldnotbeignored.With thedevelopmentofscienceand pharmacology,many drugs with aspectsof satisfactoryanalgesicefficacyand couldbe welltoleratedby patientshave been developed.And lornoxicamisone of them, which isa non-steroidalanti-inflammatorydrug (NSAID ), with analgesic, anti-infl-ammatory,andantipyreticproperties.Itseliminationhalf-time(3 to 5 hours) isle…     

Development on Adjustable Calibration Marker for Shock Wave Focus

Shock wave lithotripsy （SWL） is a treatment of choice for upper urinary stones. However, this procedure is inappropriate for obese patients because the focus is often unable to reach the target owing to the limited focal distance in shock wave source. Although treating such patients in a blast path may increase the application length of shock wave source, it＇s difficult to find this path on the lithotripter monitor. For this reason, we invented an adjustable calibration marker in order to set an effective focus in the shock wave hath.     

Study on the TCM Pathogenesis of Angiopathic Complications of Type II Diabetes

Excess production of reactive oxygen species(ROS)of mitochondrion mediated by hyperglycemia is the common pathogenesis of angiopathic complications of diabetes.TCM holds that the damp from the dysfunction of spleen.kidney and liver is the causative factor of complications of diabetes.This is similar to the mechanism of Ros resulting in angiopathic complications of diabetes.When the angiopathic complications of type II diabetes mellitus(T2DM)are difierentiated as caused by turbid damp in TCM can be explained as ROS.Since the obstruction of pathogenic damp in channels and collaterals is said to be the main pathogenesis,the treating principle should be dissolving the damp to remove the obstruction.     

Leptin, Ghrelin and TNF-α before and after Hypo-caloric Traditional Chinese Diet and Auricular Acupuncture

INTRODUCTION Obesity is a complex emergent problem, which can be possibly solved not only by the diet but also by the life style and promotion of a constant physical exercise. 1, 2 No doubt careful attentions must be given to the nutritional condition of obese people, the dietary habits, the somatic build (i.e. distribution of fat mass) and the organic functions linked to formation of the fat mass. All the parameters should be constantly monitored before, during and after a diet treatment. 3, 4, 5     

Relationship between Dysglycemia and Carotid Atherosclerosis in Tibetan Population

People with dysglycemia are at high risk for atherosclerotic diseases. This study aims at investigating the atherosclerotic vascular damage in dysglycemia and its metabolic origin in Tibetan population.     

Acupuncture Combined with Spinal Tui Na for Treatment of Primary Dysmenorrhea in 30 Cases

Objective: To observe the therapeutic effects in acupunture treatment of primary dysmenorrhea combined with spinal Tui Na, and study its mechanism. Methods: Thirty cases of the treatment group were treated by acupuncture combined with spinal Tui Na, and thirty cases in the control group were treated by routine acupuncture. Results: The total effective rate was 93.3% in the treatment group, and 73.3% in the control group, with a significant difference between the two groups (P<0.05). Conclusions: Acupuncture combined with spinal Tui Na has good prospects for treatment of primary dysmenorrhea.     

Luo Lingjie's Experience in Treating Chronic Nephropathy

In treating chronic nephropathy,Luo Lingjie,a chief physician,pays attention to regulating the balance between yin and yang,treating infection if present,and removing pathogenic factors.He prescribes gentle drugs and uses carefully strongly warming-tonifying ones,emphasizes the importance of persuading the patient to persist in treatment with medication and nurse one's health for recuperation,and is good at combined use of TCM and western medicine therapy and brings the merits of various therapies into full play,with obvious theraoeutic effects.     

Dr.Zhang Ren's Experience in the Acupuncture Treatment of Different Diseases with the Same Therapeutic Principle

Dr.Zhang Ren,the chief physician,is the chairman of Shanghai Acupuncture and Moxibustion Association.Having been engaged in medicine for about 40 years,he is experienced in treating various intractable diseases.In his long years of clinical practice,he advocates taking the TCM differentiation as the basis to seek for the acupuncture method for treatment of modern intractable diseases.The author of this essay had the fortune to follow Dr.Zhang in study.The following is a summary of Dr.Zhang's experience in the acupuncture treatment for different intractable diseases with the same therapeutic principle.