新生隐球菌的磷脂酶活力检测

目的 :评价不同孵育条件、来源和血清型的新生隐球菌分泌磷脂酶的活力情况。方法 :分别在 30℃和 37℃下用蛋黄平板法培养不同来源的 4 0株新生隐球菌菌株 ,测量其产生沉淀圈的大小 ,计算 PZ值来比较其磷脂酶活力的不同。 结果 :4 0株菌中有 39株能在其菌落周围产生明显沉淀圈 ,PZ值在 30℃和 37℃培养条件下分别为 (0 .5 2 9± 0 .12 1)和 (0 .5 2 3± 0 .14 3) ,两者差异无统计学意义 (P>0 .0 5 ) ;在 37℃培养条件下 ,血清 A、B、AD和 D型菌株的 PZ值分别为 (0 .5 4 1± 0 .116 )、(0 .5 18± 0 .0 36 )和 (0 .4 72± 0 .0 5 1) ,3组间差异均无统计学意义 (P>0 .0 5 ) ;临床株和环境株 PZ值分别为 (0 .5 0 1± 0 .0 4 9)和 (0 .5 6 5± 0 .131) ,差异有显著性意义 (P<0 .0 5 )。结论 :温度对新生隐球菌磷脂酶活力没有影响 ;不同来源的新生隐球菌菌株的磷脂酶活力差异较大 ,其中临床分离株比环境分离株磷脂酶活力强 ;而不同血清型菌株的磷脂酶活力无差别。     

细胞芯片检测新生隐球菌菌株丝氨酸蛋白酶的表达

目的:利用细胞芯片技术检测新生隐球菌菌株中丝氨酸蛋白酶的表达,初步探讨丝氨酸蛋白酶在新生隐球菌致病过程中的作用.方法:不同来源和血清型的新生隐球菌菌株36株,应用组织芯片构建仪制备菌株芯片,利用菌株细胞芯片和免疫组化技术对菌株中丝氨酸蛋白酶的表达情况进行检测.结果:丝氨酸蛋白酶在所有菌株细胞中强阳性表达率为67.0%(25/36),强阳性表达率在血清A型、血清B型、血清D/AD型分别为46.2%(6/13)、92.3%(12/13)、66.7%(4/6),在环境分离株、临床分离株及荚膜缺陷株中分别为55.6%(5/9)、82.6%(19/23)、25%(1/4).不同血清型及不同来源菌株中,以血清B型、临床分离株中的丝氨酸蛋白酶阳性表达率高(P<0.05).结论:菌株细胞芯片是检测致病性真菌样本成分的新技术;致病力强的新生隐球菌临床分离株中的丝氨酸蛋白酶表达最高,提示丝氨酸蛋白酶在临床菌株致病过程中起到主要作用.     

产黑素培养基在新生隐球菌病诊断和预后评估中的临床意义

目的:寻找新生隐球菌病的临床快速诊断方法和临床预后的评估指标.方法:用咖啡酸玉米吐温和多巴(CT)两种产黑素培养基培养128株已被明确鉴定的新生隐球菌,并以白念珠菌等作对照,25℃生长,了解显色结果.结果:非新生隐球菌及一部分新生隐球菌在两种培养基中均不显色,有12株新生隐球菌病仅在一种培养基中显色,2株临床分离株在两种培养基上均不显色.结论:产黑素培养基在新生隐球菌病的快速诊断和临床预后评估中均可起重要的作用.     

热休克预处理对化疗环境下骨髓间充质干细胞存活率的影响及机制

目的:探索热休克预处理(heat shock pretreatment,HSP)对化疗微环境下骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)的影响及其潜在机制.方法:探索HSP最佳条件,往热休克预处理BMSCs(heat shock pretreated bone marrow mesenchymal stem cells,HS-MSCs)培养基中加入顺铂,通过CCK-8检测细胞增殖率,Hoechst33342/PI测定存活率,流式细胞仪检测凋亡率.使用Western blot检测不同时间点HS-MSCs热休克蛋白70(heat shock protein 70,Hsp70)、Hsp90、自噬相关蛋白Beclin1和LC3B的表达.通过透射电镜观察自噬小体数量.结果:加入顺铂后,热休克模型组凋亡率明显低于模型组.此外,热休克模型组增殖速率较模型组增加,而表达亮蓝色/暗红色荧光的比例则低于模型组.HS-MSCs内Hsp70和Hsp90的表达随时间推移逐渐上调,并在48 h后达峰值.同时,细胞内自噬小体减少、Beclin1表达降低、LC3B Ⅱ/LC3B Ⅰ比值下降.结论:HSP能够提高化疗微环境下BMSCs的存活与抗凋亡能力,其作用可能与HSP增强Hsp70及Hsp90的表达、减弱自噬水平有关.     

两性霉素B和两性霉素B脂质体体外抗真菌的实验研究

本实验室于 1999年 1月至 2月用美国国家临床试验标准化委员会 (NCCL S)方案对上海 4家医院提供的 91株致病真菌进行了两性霉素 B(c Am B)和两性霉素 B脂质体 (Am Bi-some)的体外药敏试验 ,现将结果报告如下。1　材料和方法1.1　实验菌株　长海医院提供 :新生隐球菌 3株 ;长征医院提供 :新生隐球菌 15株 ,白念珠菌 19株 ,黄曲霉 2株 ,烟曲霉 2株 ,青霉菌 2株 ,总状毛霉 1株 ;瑞金医院提供 :新生隐球菌 4株 ,白念珠菌 17株 ,热带念珠菌 8株 ,类星型念珠菌 2株 ,黑曲霉 1株 ,克柔念珠菌 2株 ,链互隔菌 2株 ,总状毛霉 1株 ,土生念珠菌 1株…     

新生隐球菌的胞外蛋白水解酶及丝氨酸蛋白酶活性检测

目的:测定不同孵育条件、来源和血清型的新生隐球菌分泌的胞外蛋白水解酶及丝氨酸蛋白酶的活性.方法:30℃和37℃下用含有偶氮白蛋白的琼脂培养板培养不同来源及血清型的36株新生隐球菌菌株,测量其产生的廓清晕环的大小,计算CH值[菌落半径/(菌落半径 廓清晕环半径)]来比较其胞外蛋白水解酶及丝氨酸蛋白酶的活性强弱;用丝氨酸蛋白酶的特异性抑制剂观察两种酶活性改变;酶谱法观察菌株浓缩上清液中胞外蛋白水解酶活性和相对分子质量.结果:36株菌株在30℃和37℃培养条件下它们的平均CH值分别为0.558±0.170和0.575±0.169,两者间无显著差异;血清A(n=13)、B(n=13)、D/AD(n=6)型菌株各自的平均CH值分别为0.564±0.144、0.515±0.078和0.482±0.072,各组间无显著差异;临床分离株(n=23)、环境分离株(n=9)和荚膜缺陷株(n=4)各自的平均CH值分别为0.570±0.177、0.513±0.069和0.942±0.075(P＜0.05).对照组(不含抑肽酶)和抑肽酶不同浓度组(1.2、1.6 mU)的平均CH值分别为0.459±0.188, 0.975±0.287、0.733±0.252(P＜0.01).菌株浓缩液中含有复杂的胞外蛋白酶类成分.结论:新生隐球菌胞外蛋白水解酶及丝氨酸蛋白酶的活性在30℃、37℃下无显著差异;在临床分离株中比环境分离株和荚膜缺陷株强;在不同血清型菌株间无显著差别;可被丝氨酸蛋白酶抑制剂抑制.     

热休克蛋白70及其抑制剂的研究进展

热休克蛋白70(heat shock protein 70,Hsp70)是一种在应激条件下诱导表达的伴侣蛋白,能帮助细胞内新生蛋白折叠,蛋白细胞内运输、蛋自装配和降解,并能在应激状态下维持蛋白质构象.研究表明,Hsp70与许多疾病有关,如癌症、神经退行性疾病、异源移植的排斥、感染等,有望成为新的药物作用靶点.本文综述了...     

多巴培养基快速鉴定新生隐球菌产黑的研究

黑素被认为是新生隐球菌重要的毒性因子 [1 ] ,在现有的致病酵母中 ,只有新生隐球菌才能催化合成该产物 [2 ] 。对新生隐球菌黑素的研究主要集中于其致病机制以及利用其特性鉴定该菌种 ,但是否所有菌株都能产生黑素 ,或是多大程度上能产生黑素 ,以及如何更好的利用该特性为临床服务都是迫切需要解决的问题。本研究利用改良的多巴培养基对本室保存的 15 7株新生隐球菌进行显色培养 ,以期对该方法的临床意义作一客观的评价。1　材料和方法1.1　菌株　 15 7株菌株均为已明确鉴定好的新生隐球菌 ,其中 A型 35株 ,B型 2 6株 ,C型 4株 ,D型 9株 …     

热休克蛋白70对苯致小鼠骨髓细胞毒作用的研究

目的分析热应激蛋白70(heat stress or stress proteins 70,Hsp70)在苯对小鼠骨髓细胞毒性中的作用,为进一步阐明Hsp70的生物学意义和苯的防治提供科学依据.方法经过原代细胞培养后,细胞分组为: 染毒组 , 以0、2、10、50 mmol/ L浓度的苯染毒2 h ; 热应激染毒组,细胞经40℃受热1 h , 37℃恢复1 h ,再以0、2、10、50 mmol/ L浓度的苯染毒2 h.检测Hsp70表达时的光密度,使用流式细胞术观察Hsp70表达水平和小鼠骨髓细胞凋亡程度的关系.结果小鼠骨髓细胞受热应激30 min后, Hsp70轻度增加, 而应激60,90 min后, Hsp70明显升高.热应激后Hsp70表达增加;热应激组细胞除0剂量染毒组外, 其他剂量组细胞发生凋亡百分比均高于染毒组.热应激后Hsp70表达增加.结论 Hsp70的增加可以提供一定程度的毒物耐受能力或细胞保护功能,热应激后细胞Hsp70的增加不能抑制2,10 mmol/L浓度苯所致小鼠骨髓细胞凋亡的发生.     

热休克蛋白10和热休克蛋白60的表达、纯化及与线粒体DNA转录因子A相互结合

目的:表达、纯化热休克蛋白10(Hsp10)和热休克蛋白60(Hsp60),研究人类线粒体DNA转录因子A(TFAM)在体外与Hsp10,Hsp60间的相互交联作用.方法:使用Escherichia.Coli(E.coli)诱导表达含有6个组氨酸标记的Hsp10和Hsp60,镍离子鳌合树脂分离纯化后,体外分别与纯化TFAM共育,抗-TFAM抗体免疫沉淀,SDSPAGE分离蛋白,CBB探测分析.结果:Hsp10可在体外与TFAM发生免疫共沉淀,Hsp60不能.结论:Hsp10可能为TFAM-复合体组分.     

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.