抗钩虫特异性卵黄抗体的制备与鉴定

目的 制备并鉴定特异性抗十二指肠钩虫鸡卵黄免疫球蛋白(IgY),为分析抗钩虫IgY抗体的作用奠定基础.方法 制备十二指肠钩虫成虫粗抗原,经皮下和肌肉注射免疫25周产蛋海兰母鸡3次,每只鸡每次的免疫剂量为200 μg,初次免疫后28 d进行加强免疫,加强免疫间隔时间为10 d.水稀释法提取卵黄中的IgY,盐析、透析法纯化IgY,BCA法测定蛋白含量,并进行十二烷基磺酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)和Western blot分析.ELISA法动态检测抗体效价,观察抗体效价变化.结果 免疫后55 d的每个鸡蛋能提取40~55 mg IgY,经SDS-PAGE和Western blot分析,纯化的IgY有2条主带,分子量分别是35 ku和60 ku,即IgY的轻链和重链.初次免疫后10 d左右出现抗体,至免疫后50~55 d为高峰,效价达到1 ∶ 10 000以上.IgY与人血清抗体均可识别30 ku处钩虫抗原.结论 十二指肠钩虫成虫粗抗原免疫的海兰母鸡能产生高浓度、高效价、特异性的IgY抗体,为分析抗钩虫IgY抗体的作用奠定了基础.     

猪囊尾蚴细胞疫苗对仔猪先天性和获得性免疫的作用

目的观察猪囊尾蚴细胞疫苗对妊娠母猪、具有母源抗体的仔猪和正常仔猪的影响.方法通过ELISA方法检测血清抗体,以确定疫苗对不同实验猪的免疫效果.结果妊娠母猪产生的抗体可达到1：20480,并使仔猪获得的母源抗体接近有效抗体滴度1：2560,出生后适时追加免疫,短期内即可产生较高抗体水平.结论 猪囊尾蚴细胞疫苗免疫怀孕母猪后,母猪体内产生的抗体可以传递给仔猪,使仔猪获得先天性免疫力,具备抵抗猪囊尾蚴虫卵的侵袭.免疫正常仔猪产生有效抗体时间较晚.     

抗脑囊虫头节和囊壁鸡卵黄抗体的制备

鸡卵黄免疫球蛋白IgY(Yolk Immunoglobulin)是近几年研究的热点,本文把IgY用于对脑囊虫抗体的制备,为囊虫抗体的获取开辟了新思路.自新鲜病猪肉中摘取猪囊尾蚴,用TBS缓冲液浸洗并吸干水分.然后将头节和囊壁剥离,经离心及饱和硫酸铵溶液处理制成抗原.用分光光度法测定蛋白含量.将抗原与福氏完全佐剂混合制成乳浊液,免疫34周龄美国海兰特产蛋母鸡40只,分为2组,每组20只.分别进行头节和囊壁抗原的免疫,每只鸡以1ml抗原于左右背部和胸部多点注射.     

亚洲带绦虫实验感染家猪的研究

目的:观察贵州省亚洲带绦虫囊尾蚴在猪体内的发育情况。方法:用亚洲带绦虫孕节直接灌喂当地圈养的4d龄健康乳猪8头,并与健康乳猪4头作对照。于感染后40、50、60、75d分别剖检,观察猪体内囊尾蚴的分布、大小、形态及成熟情况。结果:从感染后40d起,在6头猪的肝脏表面和实质内共检获245个囊尾蚴,多分布在肝脏实质内,其大小为1.34-2.78mm×1.22-2.31mm,其余部位来找到囊尾蚴。感染后40d和50d,囊尾蚴经胆汁孵育,头节翻出率分别为78.3%和93.4%。感染后50d开始发现钙化的囊尾蚴。囊尾蚴活体观察可见头节上4个吸盘和活动的顶突,项突周围发现不完整的两圈点状结构。结论:根据囊尾蚴的分布、大小、发育时间及形态特征,进一步认定贵州省都匀市的带绦虫为亚洲带绦虫。家猪是其可能的中间宿主。     

猪囊尾蚴抗原的异质性分析

目的：分析猪囊尾蚴抗原的异质性。方法：采用间接型ELISA法，以正常猪血清（NPS）为抗原，对用猪囊尾蚴抗原（CFA）免疫的BALB／C小鼠杂交瘤细胞的培养上清液进行检测。同时设猪囊尾蚴抗原组。结果：NPS阳性21孔，阳性率24．71％（21／85）。CFA阳性24孔。结论：猪囊尾蚴抗原含有宿主的蛋白质成分。     

二例眼内猪囊尾蚴的手术治疗

目的探讨治疗眼猪囊尾蚴病的手术方法及临床疗效.方法二例猪囊尾蚴分别位于视网膜下和玻璃体后段视网膜前.前者通过眼底镜用导光纤维于囊泡外缘电凝定位,从相应的巩膜面作切口;后者则通过"玻切术"的冷冻法切除.术后辅以砒喹酮治疗.结果二例标本证实为猪囊尾蚴.术后眼部反应均较小;角膜透亮,前房基本清,玻璃体轻度混浊;视网膜仅有局部疤痕及色素沉着,少量小出血点,未见有视网膜脱离的发生.结论必须根据猪囊尾蚴不同的部位,选择不同的手术方法;药物以砒喹酮为广谱、高效;应先摘除眼内猪囊尾蚴,然后投以药物.     

抗哇巴因抗体IgY的制备及其应用

目的 制备高度特异性的抗哇巴因多克隆抗体,并将其应用于组织中内源性哇巴因的免疫学检测.方法 采用哇巴因.牛血清蛋白耦联物作为免疫原免疫母鸡.免疫后1周收集鸡蛋.鸡蛋黄采用PEG-6000方法进行纯化分离,并采用12%SDS-PAGE分析其纯度.酶联免疫吸附法检测IgY抗体效价.然后应用IgY的竞争性酶联免疫吸附法检测哇巴因水平,并将IgY应用于大鼠肾上腺组织中内源性哇巴因的检测.结果 IgY抗体效价在第二次免疫后迅速升高,最高效价达到1:10240,并持续至少4周左右.竞争性酶联免疫吸附法检测哇巴因的批内差异及批间差异分别为2.03%、2.34%.免疫组化结果显示内源性哇巴因主要分布于大鼠肾上腺皮质的网状带,髓质未见明显阳性颗粒.结论 免疫母鸡简单、快速,并且高效价抗体持续时间较长.同时本研究结果提示IgY抗体可用于常规免疫学检测.     

猪囊尾蚴抗原的异质性分析

目的：分析猪囊尾蚴抗原的异质性。方法：采用间接型ＥＬＩＳＡ法，以正常猪血清（ＮＰＳ）为抗原，对用猪囊尾蚴抗原（ＣＦＡ）免疫的ＢＡＬＢ／Ｃ小鼠杂交瘤细胞的培养上清液进行检测。同时设猪囊尾蚴抗原组。结果：ＮＰＳ阳性２１孔，阳性率２４．７１％（２１／８５）。ＣＦＡ阳性２４孔。结论：猪囊尾蚴抗原含有宿主的蛋白质成分。     

猪白细胞介素4的基因克隆及其在囊尾蚴DNA疫苗中的应用

目的：克隆猪白细胞介素4（IL－4）cDNA，观察其在抗囊尾蚴免疫中的疫苗佐剂作用。方法：通过RT－PCR法获得带有5'AUG侧翼翻译优化突变序列的猪IL－4 cDNA，测序后重组入真核表达载体pcDNA，在体外瞬时表达的基础上与囊尾蚴保护性抗原DNA疫苗联合免疫仔猪。结果：获得的猪IL－4cDNA列与献报道的完全一致，在体外瞬时表达中获得了有生物学活性的猪IL－4。其与囊尾蚴保护性抗原DNA疫苗联合应用可有效提高个体体液免疫应答水平。结论：构建了一高效表达猪IL－4的真核表达质粒，初步实验证实了其在抗囊尾蚴DNA疫苗中的佐剂效应。     

抗白念珠菌鸡卵黄抗体的制备及温度对其效价的影响

目的:制备抗白念珠菌鸡卵黄抗体(IgY)并检测其生物学性能.方法:白念珠菌标准菌株10231灭活后作为抗原免疫25周龄产蛋海蓝鸡,从水溶法提取鸡卵黄IgY抗体.用考马斯亮蓝染色法测定IgY抗体提取液的蛋白浓度.用Tris-tricine电泳测定提取蛋白的相对分子质量、纯度,Western blot确定蛋白来源.以免疫组织化学检验抗白念珠菌IgY抗体与白念珠菌结合的特异性.ELISA测定抗白念珠菌IgY抗体的抗菌效价.结果:IgY抗体液中蛋白浓度为7.9 g/L,IgY抗体的纯度达到91.5%.提取的IgY抗体与鸡血清和正常鸡IgY抗体具有同源性.提取的IgY抗体可与白念珠菌发生特异性结合. ELISA测定抗白念珠菌IgY抗体的效价达到1:12 000.结论:研究得到的IgY,产量和纯度高,效价好,体温条件下48 h效价没有降低.在体外能够特异性地与白念珠菌结合.     

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.