醛糖还原酶的分离纯化和性质分析

利用离心、盐析及ＤＥＡＥ－纤维素、羟基磷灰石、葡聚糖－Ｇ１００多步层析法将牛睾丸ＡＲ分离纯化，醛糖还原酶（ａｌｄｏｓｅｒｅｄｕｃｔａｓｅ，ＡＲ），并利用体外酶动力学方法测定其底物特异性，为寻找有效的醛糖还原酶抑制剂提供线索。结果：纯化的酶经ＳＤＳ－ｐＡＧＥ电泳鉴定示一单一蛋白条带，牛睾丸ＡＲＭｒ约为（４００００±１０００）ｕ，ＡＲ底物特异性，按１／ｋｍ大小依次排列为：Ｐ－硝基苯醛，ＤＬ－甘油醛，Ｄ－葡萄糖醛酸，Ｄ－木糖，Ｄ－半乳糖，Ｄ－葡萄糖。     

醛糖还原酶的分离纯化和性质分析

利用离心，盐析及ＤＥＡＥ－纤维素，羟基磷灰石，葡聚糖－Ｇ１００多步层析法将牛睾丸ＡＲ分离纯化，醛糖还原酶（ａｌｄｏｓｅｒｅｄｕｃａｓｅ，ＡＲ），并利用体外酶动力学方法测定其底物特异性，为寻长有效的醛糖还原酶抑制剂提供线索，结果；纯化的酶经ＳＤＳ－ＰＡＧＥ电泳鉴定示一单一蛋白条带，牛睾丸ＡＲ，Ｍｒ约为（４００００±１０００）ｕ，ＡＲ底物特异性，按１／Ｋｍ大小依次排列为：Ｐ－硝基苯醛；ＤＬ－甘油醛，Ｄ     

云南产螺旋藻多糖的分离纯化与分析

云南程海湖工业化养殖的螺旋藻（干品）经热水提醇沉，ｓｅｖａｇ法脱蛋白，透析，再经ＤＥＡＥ－ｓｅｐｈａｄｅｘ－Ａ２５与ｓｅｐｈａｄｅｘＧ－２００柱层析纯化，得螺旋藻多糖（ＰＳＰ）。用纸层析，柱层析及电泳检查纯度，表明ＰＳＰ为单一组分。ＰＳＰ经ＵＶ－３００扫描，无核酸（２６０ｎｍ）及蛋白质（２８０ｎｍ）特征吸收峰。ＩＲ－４００扫描，出现典型多糖吸收峰。经纸层析及气相色谱分析表明ＰＳＰ组成单糖为Ｌ－岩藻糖、Ｄ－甘露糖、Ｄ－半乳糖、Ｄ－葡萄糖和葡萄糖醛酸。ＰＳＰ平均分子量约为１６６００。     

受体相关蛋白在大鼠各组织中的分布

用ＳＤ大鼠肾皮质总ＲＮＡ克隆的ＲＡＰ－ｃＤＮＡ，经原核融合蛋白表达系统获得纯化的受体相关蛋白（ＲＡＰ）由此制备特异的免疫抗血清，分别提取大鼠各组织蛋白，经蛋白质聚丙酰胺凝胶电泳（ＳＤＳ－ＰＡＧＥ）分离后，转移至硝酸纤维素膜，用抗ＲＡＰ抗血清进行蛋白质印迹分析（ＷｅｓｔｅｒｎＢｌｏｔＡｎａｌｙｓｉｓ）结果显示，ＲＡＰ在大鼠肝，胰，肾皮质，肾上腺，大网膜，大脑，小脑，骨骼肌等组织内匀在不同程度的表达，     

高血糖对人红细胞醛糖还原酶活性的影响

目的研究高浓度葡萄糖对人红细胞醛糖还原酶（ａｌｄｏｓｅｒｅｄｕｃｔａｓｅ，ＡＲ）活性的影响。方法利用ＤＥＡＥ－纤维素（ＤＥ－５２）柱层析法，将正常人、糖尿病患者及体外高浓度葡萄糖孵育的红细胞醛糖还原酶部分纯化，测定其活性，并对其性质作进一步分析。结果①高浓度葡萄糖孵育的红细胞醛糖还原酶Ｋｍ值下降约６～７倍；②糖尿病红细胞醛糖还原酶活性明显高于正常人，酶活性升高程度与血糖值正相关；③高糖时醛糖还原酶不仅对多种底物的Ｋｍ值（包括葡萄糖）显著下降，而且对醛糖还原酶抑制剂（ａｌｄｏｓｅｒｅｄｕｃｔａｓｅｉｎｈｉｂｉｔｏｒｓ，ＡＲＩｓ）的抑制不敏感。结论①高血糖时ＡＲＩｓ的用量需加大，才能取得满意的疗效；②（ＤＥ－５２）柱层析法测定红细胞ＡＲ活性准确、方便，可以作为判断糖尿病慢性并发症病情及评估醛糖还原酶抑制剂疗效的一项重要指标     

螺旋藻酸性杂多糖的分离纯化和分析

螺旋藻藻粉经热水提提,乙醇沉淀,Ｓｅｖａｇ法去蛋白质,十六烷基三甲基溴化铵（ＣＴＡＢ）沉淀得酸性杂多糖,再经ＤＥＡＥ－纤维素柱层析分经纯化两次得多糖ＰＳＰ１和ＰＳＰ２。ＰＳＰ１和ＰＳＰ２经ＨＰＬＣ的Ｃａｒｂｏｈｙｄｒａｔｅ Ａｎａｌｙｓｉｓ柱层析为单一对称峰,证明为均一多糖。纸层析和硫酸－咔唑反应分析表明：ＰＳＰ１主要由Ｄ－半乳糖、Ｄ－甘露糖、葡萄糖醛酸及Ｄ－葡萄糖四种残基组成,比例为２．１：３．     

高血糖对人红细胞醛糠还原酶活性的影响

目的 研究高浓度葡萄糖对人红细胞醛糖学原酶（ａｌｄｏｓｅｒｅｄｕｃｔａｓｅＡＲ）活性的影响。方法 利用ＤＥＡＥ－纤维素（ＤＦ－５２）柱层析法，将正常人，糖尿病患者及体外高浓度葡萄糖孵育的红细胞醛糖还原酶部分纯化，测定其活性，并对其性质作进一步分析，结果 （１）高浓度葡萄糖孵育的红细胞醛糖还原酶Ｋｍ值下降约为６～７倍；（２）糖尿病红细胞醛糖还原酶活性明显高于正常人，酶活性升高程度与血糖值正相关；（３     

NON-PERFUSION COLD STORAGE METHOD FOR TOTAL THYROID-PARATHYROID PRESERVATION

ＮＯＮ－ＰＥＲＦＵＳＩＯＮＣＯＬＤＳＴＯＲＡＧＥＭＥＴＨＯＤＦＯＲＴＯＴＡＬＴＨＹＲＯＩＤ－ＰＡＲＡＴＨＹＲＯＩＤＰＲＥＳＥＲＶＡＴＩＯＮ￥ＣｈｅｎＹｕｘｉｎ（陈雨信）（Ｄｅｐｔ．ｏｆＳｕｒｇｅｒｙ，ＴａｉｓｈａｎＭｅｄｉｃａｌＣｏｌｌｅｇｅ）Ａｂｓ...     

ITP患儿血清可溶性IL—2受体水平和T淋巴细胞亚群变化

探讨特发性血小板减少性紫癜（ＩＴＰ）患儿免疫功能变化。方法采用双抗体夹心ＥＬＩＳＡ法对ＩＴＰ患儿进行了血清可溶性白细胞介素２受体（ｓＩＬ－２Ｒ）测定和ＡＰＡＡＰ法检测Ｔ淋巴细胞亚群。结果ＩＴＰ患儿血清ｓＩＬ－２Ｒ、ＣＤ８显著升高，ＣＤ４／ＣＤ８显著下降（Ｐ＜０．０１），且ｓＩＬ－２Ｒ和ＣＤ４／ＣＤ８呈负相关（ｒ＝－０．５８１，Ｐ＜０．０１）；缓解期ｓＩＬ－２Ｒ明显下降，基本接近正常水平，而ＣＤ８、ＣＤ４／ＣＤ８比值较急性期变化不明显。结论提示ｓＩＬ－２Ｒ和Ｔ淋巴细胞亚群的检测对ＩＴＰ患儿病情的监测有重要意义     

日本血吸虫多坐分子疫苗的制备及其抗原性的研究

为了制备足量、高纯度的供血吸虫保护性免疫力研究的多价分子疫苗。通过制备型ＳＤＳ－ＰＡＧＥ和电渗方法从日本血吸虫成虫分离纯化出２８ＫＤ，３１／３２ＫＤ及９７ＫＤ蛋白。经ＢＣＡ法检测蛋白含量，ＳＤＳ－ＰＡＧＥ检测蛋白纯度和分子量，Ｗｅｓｔｒｎ－ｂｌｏｔ及ＥＬＩＳＡ检测其抗原活性。结果日本血吸虫成虫２８ＫＤ，３１／３２ＫＤ及９７ＫＤ三种纯化蛋白经ＳＤＳ－ＰＡＧＥ检测均为单一条带并与所分离的蛋白分子量相符     

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.