CDK9与肾透明细胞癌临床病理特征及肿瘤侵袭的关系

目的探讨肾透明细胞癌组织CDK9的表达水平,分析其与肾透明细胞癌临床特征的关系,研究CDK9对肾细胞癌细胞迁移和侵袭能力的影响。方法选取2013年1月至2016年12月在河南科技大学第一附属医院接受手术切除治疗的肾透明细胞癌患者组织标本102例和癌旁组织标本30例作为研究对象,应用实时荧光定量PCR和Western blot分别检测CDK9的表达水平,收集102例患者性别、年龄、病理分级等临床病理资料,并分析其与CDK9表达的关系。体外应用肾细胞癌细胞(ACHN、786-O和OSRC-2)研究CDK9对肾细胞癌细胞迁移和侵袭能力的影响。结果与癌旁组织比较,CDK9 mRNA和蛋白在肾透明细胞癌组织中表达显著上调(P<0.01)。CDK9 mRNA表达水平以中位数为界分为低表达组和高表达组,两组患者的病理T分期、TNM分期和远隔转移比较,差异有统计学意义(P<0.05)。肿瘤细胞系中CDK9表达显著高于人正常肾小管上皮细胞HK-2,其中786-O及ACHN细胞中CDK9表达显著升高(P均<0.01)。体外沉默CDK9显著抑制786-O及ACHN的细胞增殖、迁移和侵袭(P<0.01)。结论 CDK9表达与肾透明细胞癌的恶性特征、癌细胞增殖、迁移和侵袭能力密切相关,有望成为肾透明细胞癌的潜在治疗靶点。     

PLOD2基因在肾透明细胞癌中的作用及与预后的关系

目的探讨PLOD2基因在肾透明细胞癌(ccRCC)组织中的表达与患者的诊断率、生存率的关系及其对细胞增殖和迁移的影响。方法收集ccRCC及对应癌旁肾组织各12例,qRT-PCR和免疫组织化学方法分别检测配对组织中PLOD2基因mRNA和蛋白的表达,分析其与患者诊断率和生存率的关系;采用siRNA技术敲减786-O和ACHN细胞中PLOD2的表达; MTT和克隆形成实验检测786-O和ACHN细胞的增殖; Transwell和流式细胞技术分别检测786-O和ACHN细胞的迁移和周期分布。结果在ccRCC组织中,PLOD2基因mRNA表达高于癌旁肾组织(P<0.01); PLOD2蛋白的阳性表达率高于癌旁肾组织[91.7%(11/12)比8.3%(1/12),P<0.001]。PLOD2高表达的ccRCC患者比低表达的生存率低(P=0.026),且具有更好的诊断率(AUC=0.636 6)。786-O和ACHN细胞中,与各自对照组相比,PLOD2敲减组的吸光度值和克隆形成数均明显减少; 786-O细胞敲减组G0/G1期细胞百分率高于其对照组[(50±4)%比(27±5)%,P<0.01]; ACHN细胞敲减组G0/G1期细胞百分率高于其对照组[(67±8)%和(59±8)%,P<0.05]; 786-O细胞敲减组迁移细胞数少于其对照组[(56±8)个比(100±12)个,P<0.01]; ACHN细胞敲减组迁移细胞数少于其对照组[(52±5)个和(143±6)个,P<0.001]。结论PLOD2基因在ccRCC组织呈高表达,与患者的诊断和预后相关; PLOD2基因下调能抑制ccRCC细胞的增殖和迁移,提示其可能在ccRCC的发生发展中具有重要作用。     

绒癌细胞靶向微泡造影剂结合特性研究

目的 探索体外靶向SonoVue微泡与绒癌细胞的结合特性、结合率及靶向微泡的稳定性,为临床进行滋养细胞肿瘤的超声定位提供依据.方法 用SonoVue微泡与兔抗人HCG抗体作用制成靶向造影剂,然后分别与绒癌细胞及子宫内膜间质细胞作用,比较各自的结合率及冲洗前后的结合率.结果 SonoVue微泡与兔抗人HCG抗体的结合率为77.6%;绒癌细胞与靶向SonoVue微泡的花环形成率为(87.8±6.3)%,明显高于子宫内膜间质细胞花环形成率的(9.4±1.7)%(P＜0.05);靶向SonoVue微泡与贴壁生长的绒癌细胞结合率为(85.4±4.7)%,冲洗后结合率为(83.1±3.8)%(P＞0.05);流式细胞仪检测绒癌细胞与靶向SonoVue微泡的结合率为81.0%.结论 在体外靶向SonoVue微泡与绒癌细胞可特异性结合,其结合力具有一定的强度,可望能抵抗毛细血管内血流生理性冲洗的破坏,实现靶向显影.     

载肝癌细胞GPC3抗体的纳米级脂质超声微泡制备及体外寻靶研究

目的：探索制备载肝癌细胞磷酯酰肌醇蛋白多糖-3（glypican-3,GPC3）抗体的新型纳米级脂质超声微泡造影剂的方法,并评价其体外对肝癌细胞的靶向结合效果。方法：以机械振荡法制备纳米级脂质微泡,并通过光学显微镜及电子显微镜检测纳米微泡大小形态、粒径分布;生物素-亲和素桥接肝癌细胞GPC3抗体与纳米级脂质超声微泡,流式细胞仪检测微泡与抗体结合效率;免疫组化检测常见3种肝癌细胞的GPC3阳性表达;微泡体外与肝癌细胞靶向结合,激光共聚焦显微镜检测寻靶效果。结果：纳米脂质微泡呈圆形,分布均匀,无明显聚集,粒径范围约350~450 nm。微泡与抗体结合效率达85.05%,人SMMC-7721、HepG2及Huh7肝癌细胞GPC3表达阳性,激光共聚焦显微镜显示载GPC3抗体纳米级脂质微泡可与肝癌细胞特异性结合。结论：本研究成功制备了具备靶向性的新型纳米级脂质超声微泡,生物素-亲和素桥接GPC3抗体可特异性结合肝癌细胞,达到靶向性识别效果,为微泡的靶向诊断及治疗研究奠定了基础。     

人前列腺癌靶向超声造影剂的制备及其体外实验研究

目的制备人前列腺细胞癌靶向超声造影剂,并对其进行鉴定,探讨其体外寻靶能力.方法静电吸附法制备免疫脂质体微泡造影剂;免疫荧光染色试验证明抗体与脂质体微泡的结合,普通光镜和荧光显微下观察微泡与前列腺癌细胞的结合情况,探讨靶向脂质体微泡对癌细胞的体外寻靶能力;同时以普通微泡作为对照.结果免疫脂质体微泡的荧光免疫染色试验为阳性;体外寻靶试验显示携带PSM(C-15)抗体的靶向脂质体微泡能较好地粘附于癌细胞周边;而对照组未见微泡与靶细胞的结合.结论采用静电吸附法成功制备了具有良好免疫活性的的人前列腺癌靶向脂质体造影剂,该造影剂在体外能高效特异性地与人前列腺癌细胞结合.     

携载CAⅨ多肽的靶向纳米泡动态监测肾癌移植瘤形成过程的实验研究

目的 应用靶向纳米泡动态监测裸鼠肾癌移植瘤的形成过程,阐明靶向纳米泡在肿瘤形成过程中的成像特征和规律,探索靶向纳米泡诊断微小肿瘤的能力.方法 建立人肾透明细胞癌皮下移植瘤裸鼠模型,于种植后第1天行二维超声检查,于第7天及第14天分别行二维、彩色多普勒血流显像(color doppler flow imaging,CDFI)和超声造影检查,定量分析移植瘤组织和癌旁组织中靶向纳米泡、空白纳米泡和微米泡的显像特点.应用免疫组织化学染色鉴定移植瘤组织和癌旁组织中碳酸酐酶Ⅸ(carbonic anhydrase Ⅸ,CA Ⅸ)的表达情况.结果 种植后第7天,二维超声和超声造影显示,种植部位形成环状移植瘤组织,中间为未吸收的细胞悬液.种植后第14天,种植部位基本形成实性皮下移植瘤组织,细胞悬液完全吸收.二维超声结合超声造影可发现直径＜1.5 mm的微小肿瘤,且靶向纳米泡在显像峰值强度、峰值降半时间、峰值降半时曲线下面积均优于空白纳米泡和微米泡(P＜0.05),免疫组织化学染色证实移植瘤组织内高表达CAⅨ,而癌旁组织未表达CAⅨ.结论 超声造影能有效监测移植瘤形成的过程,靶向纳米泡能使裸鼠肾癌皮下移植瘤组织特异性显像,有利于肾癌的早期诊断.     

BTG1在肾透明细胞癌组织中的表达及对786-O细胞株增殖和凋亡的影响

目的:探讨B细胞易位基因1(BTG1)在肾透明细胞癌组织中的表达及对肾透明细胞癌细胞株786-O增殖和凋亡的影响.方法:采用免疫组化法检测20例肾透明细胞癌及其癌旁组织中BTG1蛋白的表达.构建BTG1表达质粒及合成BTG1的干扰RNA分别转染786-O细胞,通过MTT法实验观察BTG1对786-O细胞体外增殖的影响,...     

表达抗G250人源性抗体腺病毒对肾癌细胞的影响

目的检测腺病毒Ad-G250表达的G250全长人源性抗体的生物学活性及其对肾癌细胞的影响。方法扩增纯化腺病毒Ad—G250、Ad—EGFP（对照病毒）。病毒感染LO-2细胞,ELISA法检测细胞培养液中抗体的表达量。收集病毒感染后的细胞上清液进行纯化,Westernblotting检测纯化抗体的相对分子质量及其与细胞表面G250抗原的结合能力。免疫组化法检测Ad—G250表达的抗体是否具有生物学活性。CCK-8法检测Ad—G250对肾癌Ketr-3及ACHN细胞增殖的影响。AnnexinV—PE／7-AAD法检测Ad—G250对‘肾癌Ketr-3及ACHN细胞凋亡的影响。结果病毒Ad—G250感染LO-2细胞后G250抗体的表达量随着感染天数的增加而增加。Westernblotting实验显示Ad—G250能够表达G250抗体的轻链和重链,电泳图上该抗体能使G250抗原阳性的Ketr-3和786-O细胞在相对分子质量58×10’附近出现反应条带,而G250抗原阴性的ACHN和HK-2细胞在相应位置无反应条带。细胞免疫组化实验显示Ketr-3细胞膜被染成棕黄色,而ACHN细胞未见着色。CCK-8实验结果显示,Ad—G250对Ketr-3细胞有抑制作用,而对ACHN细胞的抑制作用不明显;Ad—G250抑制Ketr-3细胞增殖存在浓度及时问依赖性。凋亡实验显示,Ad—G250诱导Ketr-3细胞凋亡作用和ACHN细胞相比差异具有统计学意义,Ad—G250诱导Kert-3凋亡作用和Ad—EGFP相比差异有统计学意义。结论腺病毒Ad—G250成功表达出具有生物学活性的人源性G250抗体,且Ad—G250可抑制表达G250抗原的肾癌细胞增殖,诱导其凋亡。     

HER2靶向聚合物超声造影微泡的制备及其体外实验

目的：制备以HER2受体为靶点的靶向聚合物微泡超声造影剂,并观测其理化特性、体外肿瘤靶向性及体外超声显像效果。方法：利用生物素-亲和素法构建HER2靶向聚合物微泡,测定其粒径、电位,评价其稳定性。在体外肿瘤靶向性实验中观察其与HER2（+）的乳腺癌细胞BT-474以及HER2（-）的乳腺癌细胞MDA-MB-231细胞结合情况。并观察靶向聚合物微泡体外超声显影效果。结果：HER2靶向聚合物微泡平均粒径为（1.78±0.37）μm,平均表面Zeta电位为（-37.40±5.74）mV。12 h内靶向聚合物微泡粒径变化差异无统计学意义（P＞0.05）。HER2靶向聚合物微泡在体外对HER2（+）肿瘤细胞具有特异靶向性。体外超声显像显示靶向聚合物超声造影剂呈细腻均匀的点状密集高回声。结论：HER2靶向聚合物微泡稳定性好,靶向能力较强,具有良好的体外超声显影效果,为进一步进行体内肿瘤靶向超声造影奠定了基础。     

SOX7在肾细胞癌中的表达及其对肾癌细胞生物学行为的影响

目的:研究Y性别决定基因7(sex determining region Y-box7,SOX7)在肾细胞癌的表达以及对肾癌细胞生物学行为的影响.方法:qRT-PCR和甲基化特异性PCR(MSP)检测SOX7在肾细胞癌组织和肾癌细胞(A498、ACHN、786-O)中的mRNA表达及甲基化状态;免疫组织化学法(immu...     

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.