阳离子交换色谱—步法制备级纯化单克隆抗体

目的：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克隆抗体方法．方法：将ＭｃＡｂ腹水用ｐＨ６．０，０．０ｌｍｏ１／ＬＰＢ透析或稀释８倍后直接上ＳＰ－４０ＨＲ阳离于交换色谱柱，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检测纯度为大于９２％，ＡＢＣ染色法测定活性为１：８００００（７．８×１０－１１ｍｏ１／Ｌ），一次纯化周期仅需５０ｍｉｎ。结论：本方法操作简单，既具有常压色谱的制备量，又具有ＨＰＬＣ的纯化速度，是获得高纯度、高活性ＩｇＧ类ＭｃＡｂ的实用纯化方法。     

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     

SPECT快速法测定肾小球滤过率的研究

受检者静脉注射９９ｍＴｃ－ＤＴＰＡ（３～８ｍｃｉ），使用ｒ相机，采集６分钟，利用Ｇａｔｅｓ法计算肾小球滤过率（ＧＦＲ）．重复性实验ｌ０例ＧＦＲ（１１～１０１．９ｍｌ／ｍｉｎ），结果两均数分别是６３．３５ｍｌ／ｍｉｎ，６２．５５ｍｌ／ｍｉｎ，Ｐ＝０．４７，两均数间均无差别，相关性很好，ｒ＝０．９８６，１５例ＧＦＲ与肌酐清除率（ＣＣｒ）比较，ＧＦＲ（８．１～１０１．７ｍｌ／ｍｉｎ）．ＣＣｒ（５．２～９７．４ｍｌ／ｍｉｎ）回归方程是ＣＣｒ＝－４．６３８＋０．９５ＧＦＲ，相关性很好，ｒ＝０．９８３     

N－单甲基左旋精氨酸对犬冠脉内皮依赖性舒张功能及内皮素－1含量的影响

作者在开胸麻醉犬心脏上，观察冠脉内灌注Ｎ－单甲基左旋精氨酸（Ｌ－ＮＭＭＡ）５ｍｇ／ｋｇ，前后冠脉血流动力学、冠脉血流储备（以阻断冠脉１５ｓ再灌注所致的反应性充血血流峰值表示）以及冠脉对不同浓度乙酰胆硷（Ａｃｈ）反应的变化，同时用放射免疫法（ＲＩＡ）测定冠脉前降支（ＬＡＤ）伴行静脉血中的内皮素－１（ＥＴ－１）含量．结果证明，Ｌ－ＮＭＭＡ灌注后心率下降，基础冠脉血流量（ＣＢＦ）下降（χ±ｓ，从２７±６ｍｌ／ｍｉｎ下降至２０±８ｍｌ／ｍｉｎ，Ｐ＜０．０５），冠脉储备下降（χ±ｓ，从９１±１９ｍｌ／ｍｉｎ下降至５０±１０ｍｌ／ｍｉｎ，Ｐ＜０．０１），平均主动脉压升高，ＥＴ－１含量明显升高（χ±ｓ，从６．５±１．０ｎｇ／Ｌ升至１５．５±３．０ｎｇ／Ｌ，Ｐ＜０．０１），Ａｃｈ引起的ＣＢＦ增加减弱（Ｐ＜０．０１）．实验结果提示，生理条件下犬冠脉一氧化氮（ＮＯ）形成对ＣＢＦ，冠脉储备有重要调节作用，同时可抑制冠脉ＥＴ－１释放．     

蝎毒素Ⅳ分离、纯化研究

取东亚钳蝎毒４ｍｌ，用０．０５ｍｏｌ／Ｌ磷酸钠缓冲液（ｐＨ７．４）稀释，在４℃条件下，１５０００ｒ／ｍｉｎ离心１５ｍｉｎ，取上清上柱（ＳｅｐｈａｄｅｘＧ－５０，长１８０ｃｍ，流速１～１．５ｍｌ／ｍｉｎ）。蝎毒经分离可得１１个组分，取ＳＶＣ－Ⅳ再次柱层析（ＳｅｐｈａｄｅｘＧ－２５，长４０ｃｍ，流速２～２．５ｍｌ／ｍｉｎ）以脱盐，经真空冷冻干燥，置－１０℃以下贮藏，生物活性可保持５ａ以上。蝎毒在聚丙烯酰胺凝胶电泳中显示１２条带以上，而ＳＶＣ－Ⅳ呈现出单一条带，相对分子质量为８５００～９０００。     

SepharoseFF凝胶柱分离参数变化对蝎毒抗癌多肽纯化的影响

目的：优选毒抗癌多肽组分Ⅲ的分离参数,方法：Ｅ 同长度的ＳｅｐｈａｒｏｓｅＦＦ阳离子交换凝胶柱及不同样品洗脱速度,比较ＡＰ－Ⅲ的分离效果。结果：当柱长为２５ｃｍ,直径为５ｃｍ,流速为１．０ｍｌ／ｍｉｎ时,可分离,ＡＰ－Ⅲ纯度为８９％;分别０．８,０．５和０．２ｍｌ／ｍｉｎ流速进行层析,可分别同６,４,３个峰,ＡＰ－Ⅲ相应的纯度为８９％,８５％,８３％。当柱长为１２．５ｃｍ,而其他条件相同时,得到     

反相高效液相色谱法测定Ca~（2＋），Mg~（2＋）－ATPase活力的方法

应用反相ＨＰＬＣ方法测定了大鼠心肌Ｃａ２＋，Ｍｇ２＋－ＡＴＰａｓｅ活力。高效液相色谱条件：Ｗａｔｅｒｓ－４８０高效液相色谱仪，ＷａｔｅｒｓＣ－１８高效反相色谱柱，流动相为０．０６６７ｍｏｌ／ＬＫＨ２ＰＯ４／Ｋ２ＨＰＯ４缓冲液（ｐＨ５．９２），流速１．５ｍｌ／ｍｉｎ，柱压３０００Ｐａ，检测波长２５４ｎｍ。方法简便，快速，灵敏度高，重复性好。     

血β_2－微球蛋白在慢性肺心病缓解期判定肾功能减退的价值

对３０例慢性肺心病缓解期患者及５０例正常人采用ＲＩＡ法测定了血清的β_２－微球蛋白（β_２－ＭＧ），结果显示：慢性肺心病缓解期患者血清β_２－ＭＧ值为３．５±０．５８ｍｇ／Ｌ，较正常人血清β_２－ＭＧ值３．２４±０．４５ｍｇ／Ｌ为高（ｔ检验，Ｐ＜０．０１），肺心病缓解期患者中有１４例（Ａ组）血清β_２－ＭＧ值为４．２９±０．３８ｍｇ／Ｌ，超过正常上限，且血ＢＵＮ６．８ｍｍｏｌ／Ｌ，Ｃｒ１３２μｍｏｌ／Ｌ，Ｃｃｒ６４ｍｌ／ｍｉｎ；另１６例（Ｂ组）血清β_２－ＭＧ值（３．１２±０．３４ｍｇ／Ｌ）正常，血ＢＵＮ５．４ｍｍｏｌ／Ｌ，Ｃｒ１１４μｍｏｌ／Ｌ，Ｃｃｒ１０２ｍｌ／ｍｉｎ，两组β_２－ＭＧ及Ｃｃｒ比较有显著差异（ｔ检验，Ｐ＜０．０１）。提示血清β_２－ＭＧ测定在慢性肺心病缓解期患者中较ＢＵＮ、Ｃｒ能更早地发现肾功能减退。     

应用血浆代用品行血液稀释的实验研究

应用改良明胶液（ＭＦＧ）和羟乙基淀粉（ＨＥＳ）两种血浆代用品，对犬进行血液稀释，比较两者对犬的血流动力不学及氧输送的影响。结果显示：当放血量１０～２０ｍｌ／ｋｇ时，ＭＦＧ的ＣＯ、ＣＩ、ＤＯ２，ＶＯ２上升（Ｐ〈０．０５），ＨＥＳ的ＣＯ，ＣＩ增加，但ＤＯ２，ＶＯ２下降（Ｐ〈０．０１）。当放血量为２０～３０ｍｌ／ｋｇ时，ＭＦＧ的ＣＯ、ＣＩ仍增加（Ｐ〈０．０５），而ＨＥＳ的ＣＯ，ＣＩ已明显下降（Ｐ〈０．０     

核素显像法测定肾小球滤过率及其临床应用的价值

用９９ｍ锝－二乙三胺五乙酸（９９ｍＴｃ－ＤＴＰＡ）显像法测定了８４例受检者的肾小球滤过率（ＧＦＲ）。１５例正常人ＧＦＲ为１１４．５±１２．９ｍｌ·ｍｉｎ－１／１．７３ｍ２；泌尿系结石组及慢性肾炎组病人ＧＦＲ分别为６７．６±３７．２ｍｌ·ｍｉｎ－１／１．７３ｍ２及５３．２±３６．５ｍｌ·ｍｉｎ－１／１．７３ｍ２，与正常组比较差异有极显著意义（Ｐ＜０．００１）；原发性高血压组病人ＧＦＲ为８８．１±４０．３ｍｌ·ｍｉｎ－１／１．７３ｍ２，较正常组有显著差异（Ｐ＜０．０５）。５３例病人的自身对比研究证实，ＧＦＲ明显比尿素氮和肌酐灵敏（Ｐ＜０．００１），且该方法在测定总肾功能的同时，可显示分肾功能及肾脏形态学方面的资料。讨论了该法测定ＧＦＲ的注意事项。     

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.     

BLOOD PRESSURE CHANGE WITH AGE IN SALT-SENSITIVE TEENAGERS

Objective To observe blood pressure change with age in salt-sensitive teenagers whose salt sensitivity were determined by repeated testing.Methods Salt sensitivity was determined through intravenous infusion of normal saline combined with volume-depletion by oral diuretic furosemide in 55 teenagers. After five years, salt sensitivity was re-examined and subject blood pressure was followed up. Blood pressure changes in salt-sensitive teenagers were compared to that of non-salt sensitive teenagers over five years.Results After 5 years, the repetition rate of salt sensitivity determined by intravenous saline loading is 92.7%. In teenagers with salt sensitivity on the baseline, both the systolic blood pressure increments and increment rates were much higher than non-salt sensitive teenagers (12.7±12.1 mmHg vs. 2.8±5.2 mmHg, P＜ 0.01; 12.2%± 12.0% vs. 2.5% ±4.4%, P＜ 0.001,respectively). There was a similar trend for diastolic blood pressure (8.4 ± 6.4 mmHg vs. 3.7 ± 6.4 mmHg, P = 0.052; 13.2% ±10.6 % vs. 6.8%± 10.1%, P = 0.053, respectively).Conclusions Salt sensitivity determined by intravenous saline loading showed good reproducibility. Blood pressure increments with age were much higher in salt-sensitive teenagers than non-salt sensitive teenagers, especially in terms of systolic blood pressure.     

安心颗粒对急诊经皮冠状动脉介入术后生活质量影响的研究

目的：评价使用安心颗粒对急诊经皮冠状动脉介入术（PPCI）术后生活质量的影响.方法：将160例接受PPCI的急性ST段抬高型心肌梗死患者随机分为安心颗粒组（术前顿服安心颗粒8.8g,术后安心颗粒4.4 g/次,每日2次）和对照组（仅接受基础药物治疗）.所有患者均服用阿司匹林、氯吡格雷和阿托伐他汀.分别在入院时、出院前1d、出院后180 d时,应用心肌梗死多维度量表（MIDAS）、中文版SF-36评价量表对患者生活质量评分.并观察术后30 d以内的出血并发症、血小板减少症发生情况.结果：入院时和出院前1d,两组患者的心肌梗死MIDAS、SF-36量表评分比较无差异（P＞0.05）;出院后180 d时,与对照组比较,安心颗粒组MIDAS、SF-36评分明显减低（P＜0.05）;组内与入院时比较,两组出院前1d、出院后180 d时,MIDAS、SF-36评分均降低（P＜0.05）.两组患者在随访期间均无大量出血、少量出血、重度和极重度血小板减少症发生,安心颗粒组有4例、对照组有7例发生不明显出血（P＞0.05）.两组发生轻度血小板减少症的患者数比较无差异（P＞0.05）.结论：PPCI使用安心颗粒,能改善急性ST段抬高型心肌梗死患者的生活质量,且不增加出血风险.     

The comparative studies of the influences of Urapidil and Nicardipine on sino-atrial node function, atrio-ventricular node function and hemodynamics

Objective:To investigate the influences of urapidil and nicardipine on rabbit sinus function,atrio-ventricular node function and hemodynamics.Methods:Thirty-two Angora's rabbits were selected and randomly divided into four groups.U1 group:urapidil 0.25 mg/kg;U2 group:urapidil 0.5 mg/kg;N1 group:nicardipine 10 μg/kg;N2 group:nicardipine 20 μg/kg.All these medicine were administrated within 30 seconds.Measurements were taken before and after the administration of urapidil or nicardipine for the following data:mean blood pressure(MAP),heart rate(HR),sino-atrial conduction time(SACT),maximal sinoatrial recovery time(SNRTmax)corrected sinus node recovery time(CSNRT),index of sinus node recovery time(SNRTI),Wenckebach A-V conduction frequency (WB),and P-R interval.Results:Significant MAP and HR changes were identified in all of the four groups before and after administration of both urapidil and nicardipine.No significant changes could be found in the rest of the parameters.Intergroup analysis showed that SACT and CSNRT of N1 and N2 groups were shorter than those of the U2 group(P＜0.01);the MAP decreased(P＜0.01)and the HR increased drastically(P＜0.01).Conclusions:Neither urapidil(0.25 mg/kg,0.5 mg/kg)nor nicardipine(10μg/kg,20μg/kg)has any significant influence on rabbit sinus function or rabbit atrio-ventricular node function.Nicardipine could be a better choice than urapidil for parafunctional sinus node patients.     

Expression of OPGmRNA and ODFmRNA in the patients of hip fracture due to osteoporosis

Objective：To investigate the gene expression of osteoprotegerin（OPG） and osteoclast differentiation factor（ODF） in the bone tissue of patients with hip fracture due to osteoporosis. Methods：OPGmRNA and ODFmRNA in the bone tissue in 50 cases of osteoporosis sufferers（over 50 years old） with hip fracture（Observer Group） and 30 cases of hip facture sufferers with no osteoporosis（Control group） were analyzed with the Semi-Quantitative RT-PCR method. Results：The mRNA expressed of ODF, OPG were both high in the patients with hip fracture. In the control group, the expression of OPG mRNA was observed, while the expression of ODF mRNA was very slight. Conclusion：Aged patients contained all signals including OPG, ODF that are essential for inducing osteoclastogenesis and promoting bone resorption.     

Dynamic Changes in Serum Estradiol and Progesterone levels in Patients of Premenstrual Syndrome with Adverse Flow of Liver-qi

Objective:To probe into the influence of changes of ovarian hormones on the pathogenesis of the specific sub-type premenstrual syndrome(PMS)and reveal partial microcosmic mechanisms of adverse flow of liver-qi.Methods:Estradiol(E2)and progesterone(P)levels in serum were determined at different phases of menstrual cycle by radioimmunoassay.Results:In the group of PMS with adverse flow of liver-qi.the secretive peak value Of E2 and P at the follicular phase significantly decreased,and the secretive peak value at the luteal phase did not come into being.Conclusions:Low E2 and P secretive peak at the follicular phase and absence of secretive peak at the luteal phase is one of the microcosmic mechanisms of PMS with adverse flow of liver-qi.One of the pathophysiologic mechanisms of specific sub-type PMS is probably the continuous low level of E2and P.     

Real-time Three-dimensional Echocardiography in Assessment of Left Ventricular and Right Ventricular Volumes

Real-time three-dimensional echocardiography (RT3DE)is a new ultrasound technique that enables dynamic threedimensional visualization and quantification of the heart in real time. Investigation of feasibility and methodology of RT3DE in determining left ventricular (LV) and right ventricular (RV) volumes, RT3DE was performed in 35 normal adults using Philips SONOS 7500 system with a 2-4 MHz matrix array transducer. The 60°×60° "pyramid" volume database was obtained and analyzed on a TomTec echo workstation. Both LV and RV volumes were calculated with four 3DE methods (i.e. apical 2, 4, 8, and 16-plane) through manually tracing ventricular endocardial borders in end diastole and end systole. Stroke volumes were then calculated. LV volume was also measured by 2DE Simpson's rule using GE VIVID 7 ultrasound machine.     

SCREENING FOR A 21-CHROMOSOME ABNORMALITY IN PREIMPLANTED EMBRYOS OF ELDERLY WOMEN

Increasing maternal age is the only etiological factor unequivocally linked to Down's syndrome in humans. The occurrence rate of newborns with Down's syndrome is about 1/220 in women over 35 years old. However, the occurrence rate in embryos fertilized in vitro, of the elder woman is unclear. Using FISH we screened the number of chromosome 21 in preimplanted embryos of 5 elderly women (average age, 38.4 years) to study the feasibility and necessity of screening trisomy 21 in embryos in patients over 35 years old at the in vitro fertilization (IVF) center.     

Scientific principles and rigorous processes should be followed in developing clinical guidelines for therapeutic interventions of integrative medicine

A clinical guideline for the therapeutic interventions of integrative medicine may be defined as a written document which states a series of recommendations on therapeutic interventions of integrative medicine for a special disease or condition. The guideline may provide assistance to medical professionals in making clinical decisions aimed at improving the clinical outcome of patients and reducing the costs of medical care（~＇4~. Recommendations issued by a guideline should be based on the best available evidence in both Western and Chinese medicine. For fulfilling this purpose, the development of clinical guidelines for therapeutic interventions in the field of integrative medicine should follow scientific principles and undergo a rigorous processes.