抗日本血吸虫单克隆抗体制备及其靶抗原性质的研究

为建立两株抗日本血吸虫单克隆抗体－ＣＡ－７、ＣＡ－８，（均为ＩｇＭ型）。用单抗ＤＯＴ－ＥＬＩＳＡ方法可检测出血吸虫感染兔血清中的靶抗原。ＣＡ－７的靶抗原存在于虫卵、尾蚴中；ＣＡ－８的靶抗原存在于虫卵、尾蚴和成虫中。用亲和导析的方法分离纯化了这两株单抗的靶抗原，实验结果表明ＣＡ－７和ＣＡ－８的靶抗原是中性糖含量分别为２３．３％和１４．７％，Ｍｒ为２２０×１０＾３和２００×１０＾３的糖蛋白，这两种抗原     

人皮肤成纤维细胞纤连蛋白的纯化及其N-糖链结构

研究人皮肤成纤维细胞纤连蛋白及其片段 Ｎ－糖链结构。用明胶－Ｓｅｐｈａｒｏｓｅ ４Ｂ亲和柱纯化培养的人皮肤成纤维细胞所分泌的纤连蛋白（Ｆｎ），用胰糜蛋白酶限制性水解，获得明胶结合片段和肝素结合片段；用肼化的凝集素与经过碘酸氧化后的辣根过氧化物酶（ＨＲＰ）缩合的方法制备 ４种 ＨＲＰ－凝集素（Ｃｏｎ Ａ， ＤＳＡ，ＷＧＡ，ＬＣＡ）探针，用Ｗｅｓｔｅｒｎ Ｂｌｏｔ法检测明胶结合片段（Ｍｒ４３ × １０３）、肝素结合片段（Ｍｒ３０ ×１０３，２５ ×ｌ０３）在切除唾液酸及平分型 Ｎ－乙酰氨基葡萄糖（ＧＩｃＮＡｃ）前后的 Ｎ－糖链结构。结果： Ｆｎ的 Ｎ－糖链主要存在于 Ｍｒ分别为４３ × １０３、３０ ×１０３、２５ ×１０３的３个片段上，其结构为二、三（主要是Ｃ２Ｃ２６）或四天线的复杂型糖链；糖链末端连有唾液酸（ＳＡ）；都含有核心岩藻糖（ｃ－Ｆｕｃ；平分型 ＧｌｃＮＡｃ只存在于 ４３ × １０３片段所连的糖链中，而且很可能与 Ｃ—Ｆｕｃ共同存在于二天线或Ｃ２Ｃ２６三天线精链上；Ｆｎ的糖链中不含有Ｎ－乙酰乳糖胺重复顺序。结论：在Ｆｎ不同片段上均含有复杂型Ｎ糖链，并存在核心岩藻糖结构，平分型ＧｌｃＮＡｃ只存在于４３ ×１０３片段上。     

合成衣原体属特异LPS决定簇糖结合物的抗原性分析

合成衣原体属特异脂多糖（ＬＰＳ）的四糖链α－２－酮基－３－脱氧辛酸（ＫＤＯ）（２→８）－α－ＫＤＯ（２→４）－α－ＫＤＯ（２→６）－β－ＧｌｃＮＡｃ与牛血清白蛋白（ＢＳＡ）共价结合，制成糖－ＢＳＡ结合物抗原，用其免疫小鼠所制备的单克隆抗体（ＭｃＡｂ）经间接血凝试验、血凝抑制试验、ＥＬＩＳＡ、免疫荧光试验和免疫印渍等试验分析，结果把这些ＭｃＡｂ分为两组：一组与衣原体ＬＰＳ和Ｒｅ型ＬＰＳ有交叉反应；另一组为衣原体特异性抗体，其中ＫＤＯ３特异抗体与衣原体原体ＭｃＡｂ有共同决定簇特异性，但亲和力却高于后者１００多倍。表明合成糖－ＢＳＡ结合物与衣原体ＬＰＳ有相似的抗原性。     

血吸虫重组BCG—Sj26GST疫苗对小鼠肝脏虫卵肉芽肿形成？…

目的 检测血吸虫重组ＢＣＧ－Ｓｊ２６ＧＳＴ疫苗对小鼠肝虫卵肉芽肿细胞内ＴＮＦ－ｍＲＮＡ表达的影响。方法 用１０＾６ＣＦＵ疫苗皮下注射和腹腔注射分别免疫ＢＡＬＢ／Ｃ鼠,免疫后８Ｗ用日本血吸虫尾蚴进行攻击感染,感染后６Ｗ剖杀小鼠,分离肝脏,运用原位杂交技术检测ＴＮＦ－αｍＲＮＡ表达。结果 疫苗免疫,尾蚴攻击后小鼠肝虫卵肉芽肿细胞内ＴＮＦ－αｍＲＮＡ表达显著加强。结论 血吸虫重组ＢＣＧ－Ｓｊ２６ＧＳＴ疫     

血吸虫重组BCG-Sj26GST疫苗对小鼠肝脏虫卵肉芽肿形成过程中TNF-αmRNA表达的影响

目的 检测血吸虫重组ＢＣＧ－Ｓｊ２６ＧＳＴ疫苗对小鼠肝虫卵肉芽肿细胞内ＴＮＦ－ｍＲＮＡ表达的影响。方法 用１０～６ＣＦＵ疫苗皮下注射和腹腔注射分别免疫ＢＡＬＢ／Ｃ鼠,免疫后８Ｗ用日本血吸虫尾蚴进行攻击感染,感染后６Ｗ剖杀小鼠,分离肝脏,运用原位杂交技术检测ＴＮＦ－α ｍＲＮＡ表达。结果 疫苗免疫,尾蚴攻击后小鼠肝虫卵肉芽肿细胞内ＴＮＦ－α ｍＲＮＡ表达显著加强。结论 血吸虫重组ＢＣＧ－Ｓｊ２６ＧＳＴ疫苗能促进宿主Ｔｈ１反应,加强宿主抗血吸虫感染的保护性免疫力。     

血吸虫重组BCG—Sj26GST疫苗接种后保护力的观察

为了探讨血吸虫重组ＢＣＧ－Ｓｊ２６ＧＳＴ疫苗免疫鼠后对尾蚴攻击感染的保护性作用，采用１０＾６ＣＦＵ和１０＾６ＣＦＵ重组ＢＣＧ－Ｓｊ２６ＧＳＴ疫苗皮下接种ＢＡＬＢ／Ｃ鼠，接种后８周用日本血吸虫尾蚴进行攻击感染，感染后６周剖杀小鼠，计算减虫率、减卵率，同时进行虫卵肉芽肿周长和面积测量，以及抗体水平和抗原水平测定，同时设有ＰＢＳ对照组。结果表明重组ＢＣＧＴ－Ｓｊ２６ＧＳＴ疫苗免疫小鼠后，１０＾６ＣＦＵ、     

PAI－1糖基化位点突变体的构建及其在哺乳动物细胞中表达

Ⅰ型纤溶酶原激活物抑制剂（ＰＡＩ－１）是一种Ｍｒ为５０×１０３的单链糖蛋白，有３７９个氨基酸残基，３个Ｎ型糖基化位点。构建ＰＡＩ－１糖基化突变体，以便研究糖链的功能。用寡核苷酸定位突变技术将３个糖基化位点２０９，２６５，３２９位进行突变，把３个糖基化位点都发生了突变的ＰＡＩ－１ｃＤＮＡ组装到真核表达载体ｐＳＶ２中，得到真核表达质粒ｐＺＨ－ｐ１－Ｍ３Ｅ；在二氢叶酸还原酶缺陷型中国仓鼠卵巢细胞（ＣＨＯｄｈｆｒ－）中进行短暂表达，用发色底物法和夹心ＥＬＩＳＡ方法检测培养液中ＰＡＩ－１的活性和含量。结果：糖基化位点突变的ＰＡＩ－１能在ＣＨＯ细胞中表达，但表达水平及活性较低。非糖基化ＰＡＩ－１的活性和抗原分别为４．３４ＩＵ／ｍｌ和３．１５μｇ／Ｌ结论：用寡核苷酸定位突变方法获得了ＰＡＩ－１糖基化突变体，并且在ＣＨＯ细胞中得到表达。     

掌叶半夏凝集素A的分离纯化及分析

首次从中草药掌叶半夏（ＰｔｎｅｌｌｉａｐｅｄａｔｉｓｅｃｔａＳｃｈｏｔｔ）中通过ＳｅｐｈａｄｅｘＧ－１００分子筛层析、ＤＥ－３２离子交换层析和制备电泳分离纯化出一种对兔红血球有血凝活性的凝集素，称之为掌叶半夏凝集素Ａ（ＰｉｎｅｌｌｉａｐｅｄｔａｉｓｅｃｔａｌｅｃｔｉｎＡ，ＰＰＬ－Ａ）。在ＰＡＧＥ、ＳＤＳ－ＰＡＧＥ和等电聚焦电泳上ＰＰＬ－Ａ均显示出一条蛋白质着色带，经ＳＤＳ－ＰＡＧＥ测定其Ｍｒ为１６．３×１０３，用等电聚焦电泳测得其等电点为５．８，经ＤＮＳ－Ｃｌ分析法分析其氨基末端为丙氨酸。对所含的１８种氨基酸作了定量分析。通过Ｓｃｈｉｆｆ试剂电泳染色法鉴定ＰＰＬ－Ａ为糖蛋白，用酶标凝集素结合法分析了糖链的结构，糖链部分含有甘露糖，岩藻糖和乙酰氨基葡萄糖。     

单克隆抗体检测阳性钉螺内血吸虫抗原的初探

在筛选ＭｃＡｂ与受检钉螺样本的基础上，建立了ＭｃＡｂ－ＥＬＩＳＡ，检测钉螺体内日本血吸虫抗原，每只受检钉螺，事先镜检作为对照，感血吸虫钉螺３６只，血吸虫抗原阳性检出率９７．２２％（３５／３６）。合并感其他寄生虫钉螺７只，其阳性检出率为１００％（７／７）。感染其他寄生虫钉螺８只，无感染钉螺２２只，血吸虫抗原阴性符合率为１００％。以上结果表明，ＭｃＡｂ－ＥＬＩＳＡ与镜检法所得的结果是一致的，该方法将为     

6A8 cDNA编码一种α-甘露糖苷酶

目的 探讨６Ａ８ ｃＤＮＡ编码的蛋白质的性质。方法 依据Ｃｏｎ Ａ的配体是基露糖,而α－甘露糖苷酶的作用是剪细胞糖蛋白聚糖中苷露糖的原理,采用细胞免疫荧光染色法以及激光共聚焦显微镜观察经转染６Ａ８ｃＤＮＡ正义或反义重组表达载体后,细胞结合ＣｏｎＡ的强弱,验证６Ａ８ｃＤＮＡ编码的蛋白质是否为一种α－甘露－糖苷酶。结果 ＣＯＳ－６ 人鼻咽癌细胞株ＣＮＥ－２Ｌ２转染ｐＲｃ／ＣＭＶ－正交６Ａ８ｃＤＮＡ后与     

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.     

The clinicopathological and immuohistochemical analysis of solid-pseudopapillary tumor of the pancreas： report of 9 cases

Objective：To investigate the clinical features, pathological characteristics and immunophenotype of solid-pseudopapillary tumor of the pancreas（SPTP）. Methods：Nine surgically treated cases of SPTP were retrospectively reviewed. Hematoxylin and Eosin（HE） staining and immunohistochemical staining were used to analyze all cases, and the general clinical data was collected. Results：Six patients were asymptomatic except for a palpable mass. Two patients complained of vague-epigastric pain. One patient appeared jaundice. The tumor was encapsulated and solid tissues alternately with cystic tissues. Histologically, the histological structure of solid portion was pseudopapillary with a fibrovascular core. Tumor cells were uniform and medium-sized which were arranged in sheets ets or nests or pseudopapillary patterns. Immunohistochemical studies demonstrated that SPTP proved positive in vimentin（9/9 cases）, AAT（9/9 cases）, NSE（9/9 cases）, ACT（7/9 cases）, CK20（2/9 cases）, CgA（1/9 cases）, S-100（3/gcases）, PR（4/gcases）, Syn（3/9 cases） and CD56（5/9cases）, negative in CEA and ER. Conclusion：SPTP is a tumor predominantly occurring in young women frequently without special symptoms. This tumor has various characteristical histological patterns with different immunophenotype.     

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.