人肺腺癌细胞在裸小鼠体内脑、骨、肌肉转移模型的建立

我们将人肺隙癌细胞系Anip-9代腹水型细胞，经裸小鼠体内传代和体外培养反复交替传代筛选．获得了Anip系列的亚系．给以代号(815、926．973)．然后对Anip-19代腹水型癌细胞和Anip系列的亚系进行转移能力比较。结果Anip系列亚系中Anip 973转移能力最强，并出现脑、骨、肌肉转移。而AniP 19代腹水型细胞未出现脑．骨．肌肉转移，肺转移率也低于Anip 973．     

人成骨肉瘤高转移性细胞亚群的筛选及生物学特性的观察

目的　从低转移性人成骨肉瘤细胞系 SOSP- 96 0 7中筛选出高转移亚群 ,并且建立人成骨肉瘤裸鼠转移模型 .方法　采用裸鼠腹腔传代的方法 ,获得腹水型自发性肺转移细胞 ,经体外培养和体内再次接种后获得高转移亚群 .应用细胞计数法、染色体显示法、流式细胞术及裸鼠体内实验法研究细胞的生物学特性 .结果　筛选出的高转移亚群 SOSP- M在转移分析实验中达到10 0 %肺转移率 ,除肺转移外还出现其他器官的转移 .其染色体为 5 0余条 ,保持人类染色体核型 .细胞周期中 S期细胞所占比例为 48.5 % . SOSP- M的细胞形态、染色体数目、体外及体内…     

不同剂量H22肝癌细胞株对C57BL/6J小鼠原位肝癌造模的影响研究

目的:比较不同剂量H22传代小鼠腹水细胞构建原位肝癌移植模型小鼠的成瘤率、腹水率及存活率,以期构建和完善该模型。方法:115只C57BL6/J小鼠随机分为三组(10μL组、20μL组、30μL组),选取浓度为1×1013/m L的H22传代小鼠腹水细胞沿肝长轴直接注射接种到肝脏实质内,每组分别于第10、20 d随机选取15只给予麻醉后脱颈椎处死,肝脏称重,测量瘤体体积及重量,观察肝脏外观及病理学改变,每组预留5只观察其存亡天数,计算成瘤率、腹水率及存活率。结果:115只模型小鼠有114只均可见肿瘤,成瘤率99%;10μL组腹水率7%,20μL组腹水率46%,30μL组腹水率63%。结论:使用10μL浓度为1×1013/m L的H22传代小鼠腹水细胞悬液建立小鼠原位肝癌模型,其成瘤率高,腹水率低,存活时间较长,是较为理想的肝癌小鼠模型。     

人源性前列腺癌动物模型的初步建立

目的 建立中国人同一起源肿瘤不同恶性潜能前列腺癌动物模型,为研究前列腺癌转移进展机制及激素抵抗的发生机制提供理想的动物模型.方法 采用组织块外科原位移植法将人前列腺癌组织分别移植到虚拟去势组及去势组雄性裸小鼠前列腺部,成瘤后进行鼠间传代移植,选取虚拟去势组原位移植瘤、移植淋巴结转移瘤、去势组第4代淋巴结转移瘤体外培养建立细胞系,应用Boyden Chamber细胞运动实验检测细胞迁徙转移能力,描绘细胞生长曲线分析细胞增殖及激素依赖性,裸小鼠皮下异种移植瘤模型检测成瘤率、肿瘤湿重及浸润范围验证不同代移植瘤细胞在裸小鼠异种移植的生长状态.结果 第1代虚拟去势组成瘤率30％,无淋巴结转移,取移植瘤反复原位传代移植第3代成瘤率50％,盆腔淋巴结转移率40％.去势组原代移植以及来自虚拟去势组的原位移植瘤反复原位移植均未见移植瘤,源自虚拟去势组淋巴结转移瘤组织反复原位传代移植至第4代,成瘤率33％,其中一只可见盆腔淋巴结转移.三种细胞系细胞生长、迁徙以及体外裸小鼠成瘤率均有显著差别（P＜0.05）.结论 裸小鼠前列腺癌原位移植反复传代可增强肿瘤成瘤率、转移力等恶性指标.以裸小鼠淋巴结转移瘤接种去势裸小鼠可筛选出激素非依赖性异种移植瘤及转移瘤.     

两个不同转移力小鼠肝癌细胞系的细胞遗传学比较

Hca-F25/CL-16A_3(以下简称16A_3)和Hca-F25/CL-A_2(以下简称A_2)是两个来自共同祖先Hca-F25/L的具特异性淋巴道转移能力的小鼠腹水型肝癌细胞系。转移率前者为78.6～89.4％,后者为15～15.4％。通过细胞遗传学某些指标的观察,我们发现两系细胞存在着差异。核型分析发现,A_2系众数为42,标记染色体有四种,各一条;16A_3系众数为81和84,标记染色体也有四种,各双份,显带带型与A_2系之标记染色体完全相同。对银染核仁组织者区的观察发现:问期细胞的Ag-NORs数平均为:A_2 44个、16A_368个;中期细胞Ag-NORs平均数为:A_28.4个、16A_3 13.8个,高转移力的16_3A细胞明显高于低转移力的A_2细胞。结果提示:16A_3细胞可能是在祖先群体中由A_2细胞经核内复制而来;一些基因(如γDNA)的倍增和活性表达与16A_3高转移能力的形成可能有关。     

胰酶和Ⅳ型胶原酶在人胚胎干细胞传代中的作用特点

目的对比人胚胎干细胞(hESC)培养过程中不同的酶消化细胞的特点,探讨适合hESC长期体外培养的消化传代方法。方法将正常生长在小鼠胚胎成纤维细胞(MEF)滋养层上的hESC分别用0.05%胰酶-EDTA和Ⅳ型胶原酶消化,重新接种,了解两种酶消化后的细胞存活率、克隆形成数量、细胞产量、冻存-复苏后细胞的存活率以及长期传代后细胞核型情况。结果用0.05%胰酶-EDTA消化传代后形成的克隆大小均一,每代第3d可获得碱性磷酸酶(AP)阳性克隆数(21.2±3.5)个/10倍视野,每次传代时hES细胞产量可达(4.56±1.29)×105/cm2,传代时细胞存活率高达95.60%,经冻存-复苏后也有81.20%细胞存活率。用Ⅳ型胶原酶消化组每代仅可获得低浓度胰酶消化组AP阳性克隆数的一半,克隆大小不均,细胞产量也比低浓度胰酶消化组少,传代时细胞存活率和复苏后细胞存活率和低浓度胰酶组相似。分别用两种酶处理传代十余代,核型均无异常。结论两种酶消化均适用于hESC培养,二者作用特点不完全相同,分别适用于不同的实验需要。     

小鼠神经干细胞的分离和培养及其鉴定

目的：探讨体外分离和培养及鉴定小鼠神经干细胞,为相关的实验研究奠定基础。方法：利用神经干细胞条件培养基和单细胞克隆技术,从胚胎小鼠大脑皮质分离并体外培养胚胎期小鼠大脑皮质细胞,连续稳定传代20代后,以免疫荧光细胞化学法检测克隆细胞Nestin和分化后细胞中胶质原纤维酸性蛋白、β-tublin和半乳糖苷酶的表达。结果：从小鼠大脑皮层分离的细胞群具有连续形成克隆能力,其Nestin表达阳性。分化后的细胞可表达神经元、星形胶质细胞和寡突胶质细胞的特异性抗原。结论：成功分离并获得了小鼠皮层神经干细胞,该细胞可连续稳定传代,具备多向分化潜能,可用于进一步的实验研究。     

介绍一种使悬浮的肝癌细胞贴壁的方法

肝癌是我国常见的恶性肿瘤之一。在过去几年中,本室凌茂英等建立了615小鼠腹水型肝癌Hca-F25/L淋巴道转移体外培养细胞系,分离出比较稳定的高、低不同转移能力克隆细胞,并对其生物学特性进行了比较研究,在肝癌的病因和     

小鼠腹水型肝癌高淋巴道转移瘤株Hca-F的再克隆技术

[目的 ]探讨小鼠腹水型肝癌高淋巴道转移瘤株Hca -F的再克隆技术。 [方法 ]采用有限稀释法对已使用多年的小鼠腹水型肝癌高淋巴道转移瘤株Hca -F进行再克隆 ;6 15小鼠皮下接种 ,检测再克隆瘤株淋巴结转移能力。 [结果 ]共分离出 3个不同的瘤株 ,Hca -F - 2 - 2 4c6 ,Hca -F -3-d4 ,Hca -F - 3-a4 ,流式细胞仪检测证明 3株瘤细胞均为单克隆细胞。动物实验证明三株瘤细胞淋巴结转移率分别为 95 .5 % ,87% ,4 2 .5 %。 [结论 ]有限稀释法操作简单 ,克隆的形成率高     

人类晶体上皮细胞的体外培养与观察

目的：建立人类晶体上皮细胞培养的简便方法并观察原代和传代细胞的生物学特征和组织学变化。方法：将人工晶体植入术中取下的前囊膜分割成小碎片，吸管转移至培养瓶底部进行原代培养，7—10d后常规方法进行传代培养，采用相差显微镜观察活体细胞的增殖活动和形态学变化。结果：人类晶体上皮细胞在体外培养时可以存活并传代，但是其生存能力与供体年龄有关。传代后期细胞发生纤维化改变。结论：本方法简便，有效。可用于实验研究。     

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.