乙酰肝素酶反义寡核苷酸对裸鼠人肺癌细胞皮下移植瘤的抑制作用

目的 探讨乙酰肝素酶反义寡核苷酸(HPSE ASODN)对人肺癌A549细胞裸鼠皮下移植瘤的抑制作用.方法 设计合成互补于HPSE mRNA起始密码区的HPSE ASODN,以阳离子脂质体Lipofectamine包埋后转染人肺癌A549细胞进行培养.实验分ASODN组、脂质体组和对照组,采用Western-blotting检测各组A549细胞HPSE蛋白的表达;将反义寡核苷酸(ASODN)转染成功的人肺癌A549细胞注射于裸鼠皮下复制移植瘤模型(ASODN组),绘制移植瘤生长曲线,以免疫组织化学方法检测移植瘤肿瘤微血管密度(MVD),比较3组间肿瘤体积和MVD的差异.结果 与对照组和脂质体组比较,ASODN组A549细胞HPSE蛋白的表达受到明显抑制,差异有统计学意义(P ＜0.01);ASODN组裸鼠成瘤时间晚,移植瘤体积明显小于脂质体组和对照组,差异有统计学意义(P＜0.01).ASODN组、脂质体组和对照组移植瘤的MVD计数分别为(13.5±1.8)、(24.3±2.5)和(24.7±2.6),与脂质体组、对照组比较,ASODN组MVD计数明显降低,差异有统计学意义(P＜0.01).结论 HPSE ASODN明显抑制棵鼠人肺癌移植瘤的生长和血管生成,有希望成为一种有效的肺癌基因治疗方法.     

MMP-9ASODN对肺腺癌A549细胞增殖与凋亡的影响及其机制初步探讨

目的 探讨基质金属蛋白酶-9反义寡核苷酸(MMP-9ASODN)对肺腺癌A549细胞增殖与凋亡的影响,并初步探讨其机制.方法 MTT法检测MMP-9ASODN转染对A549细胞生长增殖的影响;流式细胞术检测MMP-9ASODN转染对A549细胞周期和凋亡比率的影响;RT-PCR法与Western blot法分别检测MMP-9ASODN转染对A549细胞内MMP-9mRNA及蛋白表达的影响.结果 在一定范围内,MMP-9ASODN对A549细胞生长抑制呈浓度和时间依赖性,反叉寡核苷酸浓度为600 nmo/L作用48 h时其抑制作用最明显;MMP-9ASODN转染A549细胞48 h后G1期细胞数明显增多,S期细胞数明显减少,G2期细胞数则无明显变化,其凋亡百分比明显升高;与对照组比较差异有统计学意义(P＜0.01);细胞内MMP-9mRNA及其蛋白的相对表达量明显低于对照组(P＜0.01).结论 MMP-9ASODN转染能够有效能够抑制肺腺癌A549细胞的增殖同时诱导其凋亡,其机制可能是通过下调MMP-9mRNA及蛋白的表达.     

MMP-2反义寡核苷酸对膀胱癌荷瘤裸鼠的治疗作用

目的 探讨基质金属蛋白酶2(MMP-2)反义寡核苷酸对人膀胱癌裸鼠异体移植瘤生长的抑制作用.方法 制备膀胱癌裸鼠移植瘤模型18只,随机分为3组:MMP-2反义寡核苷酸(ASODN)治疗组、MMP-2正义寡核苷酸(SODN)治疗组及对照组,每组6只.待瘤结节直径≥5mm后,分别在肿瘤细胞接种部位周围及中心皮下注射反义寡核苷酸/阳离子脂质体复合物、正义寡核苷酸/阳离子脂质体复合物和生理盐水.每周2次,连续4周,断颈处死裸鼠,计算肿瘤体积,称量瘤重.常规切片,HE染色,观察组织学形态并进行病理学评估.结果 与对照组瘤重(7.49±0.53)g比较,ASODN组瘤重(4.18±0.53)g明显降低( P<0.01);ASODN组、正义寡核苷酸(SODN)组抑瘤率分别为44.19%、8.41%( P<0.01); ASODN组瘤体病理学特征改善.结论 MMP-2反义寡核苷酸能抑制裸鼠体内移植瘤生长,通过反义寡核苷酸下调MMP-2的表达,有效逆转肿瘤的恶性表型,为膀胱癌的基因治疗提供了实验依据.     

Effects of Survivin antisense oligonucleotide on molecular mechanism of inducing human gastric transplantable tumor apoptosis

目的 研究survivin反义寡核苷酸(antisense oligodeoxynucleotide,ASODN)对人胃癌细胞裸鼠移植瘤生长的影响,并探讨其诱导人胃癌移植瘤凋亡的分子生物学机制.方法 建立SGC-7901胃癌细胞裸鼠皮下移植瘤模型,随机分为4组,即survivin-ASODN+脂质体组、survivin-ASODN组、脂质体组和空白对照组(蒸馏水).瘤体内隔日给药,停药后48小时处死裸鼠,剥离瘤组织,称取瘤重,计算抑瘤率.western-blot法检测各组凋亡相关蛋白survivin、Caspase-3蛋白的表达.结果 survivin- ASODN+脂质体组的移植瘤生长速度明显减慢,其抑瘤率为:33.19％.survivin-ASODN+脂质体组的survivin蛋白表达量明显低于对照组(P＜0.05),Caspase-3蛋白表达高于对照组(P＜0.05).结论 survivin反义寡核苷酸转染胃癌移植瘤能够明显抑制肿瘤生长,其主要机制与降低survivin蛋白的表达,诱导Caspase-3高表达,从而促进肿瘤细胞凋亡有关.     

ILK反义寡核苷酸对人上皮性卵巢癌裸鼠皮下移植瘤的治疗作用

目的 探讨整合素连接激酶(integrin linked kinase,ILK)反义寡核苷酸(antisense oligonucleotide,ASODN)对人卵巢癌裸鼠皮下移植瘤的治疗作用.方法 将人卵巢癌H0-8910细胞株接种于裸鼠皮下,建立人卵巢癌裸鼠模型,并将裸鼠随机分为对照组和治疗组,每次腹腔注射治疗后称量裸鼠体重、肿瘤体积及重量,并计算抑瘤率.同时,分别用脂质体和脂质体-ILK-ASODN转染H0-8910细胞,用WST-1法检测转染后两组细胞体外的增殖情况,用流式细胞术检测细胞凋亡情况.结果 治疗组裸鼠肿瘤体积及重量增长速度受到明显抑制,与对照组比较差异有统计学意义(P＜0.01).转染了ILK-ASODN的治疗组细胞增殖受到显著抑制,凋亡率明显增高,与对照组比较差异均具有极显著意义(P＜0.01).结论 ILK-ASODN在体外能抑制细胞生长、促进细胞凋亡;在体内能明显地抑制肿瘤生长,对人卵巢癌裸鼠移植瘤具有一定的治疗作用.     

人端粒酶RNA反义寡核苷酸靶向治疗对裸鼠食管癌移植瘤的作用

目的:观察人端粒酶RNA(hTR)硫代修饰性反义寡核苷酸(ASODN)对裸鼠食管癌移植瘤的治疗作用.方法:10只裸鼠随机等分为ASODN组和对照组.2组裸鼠均给予EC9706细胞悬液皮下注射,当裸鼠食管癌移植瘤长至50 mm3时,ASODN组进行每24 h 1次、连续7 d的ASODN治疗,对照组给予PBS.分别于治疗后4 d、8 d测量2组肿瘤体积,第8 d取裸鼠肿瘤组织常规病理切片观察形态学改变,TUNEL染色观察癌细胞凋亡情况.结果:治疗后第4 d,ASODN组及对照组食管癌移植瘤体积(V/mm3)分别为55.8±12.0和98.5±18.2(P＜0.01),第8 d分别为64.8±21.6和173.0±21.2(P＜0.01).TUNEL染色显示,ASODN组肿瘤组织中可见大量呈片状分布的凋亡细胞,对照组仅见少量散在的凋亡细胞.对照组肿瘤细胞异型性较ASODN组明显.结论:hTR反义寡核苷酸可有效抑制裸鼠食管癌移植瘤的生长及恶性表型的产生.     

Survivin反义寡核苷酸治疗耐顺铂人肺腺癌细胞裸鼠移植瘤的实验研究

目的:探讨以存活素基因(survivin)为靶点反义基因治疗耐顺铂(CDDP)人肺腺癌移植瘤的可行性。方法:常规体外培养A549/CDDP细胞,采用皮下注射法建立移植瘤裸鼠动物模型。以脂质体包裹的survivin反义寡核苷酸(ASODN)直接移植瘤内注射,观察肿瘤生长情况,计算抑瘤率和肿瘤生长指数。采用逆转录-聚合酶链反应(RT-PCR)和免疫组织化学分别检测survivinmRNA和蛋白的表达。结果:单用ASODN组的抑瘤率和肿瘤生长指数分别为35.4%和4.230±0.886,和空白对照组比较,抑瘤率较高,肿瘤生长减缓(P<0.05)。而ASODN联合CDDP组的抑瘤率和肿瘤生长指数则分别为63.7%和1.700±0.436,与CDDP组、Lip与CDDP联用组及单用ASODN组比较均有统计学差异(P<0.05)。RT-PCR半定量和免疫组织化学显示,ASODN组和ASODN联合CDDP组肿瘤组织中survivinmRNA及其蛋白表达明显下调(P<0.05)。结论:Survivin反义寡核苷酸可以抑制耐顺铂人肺腺癌移植瘤的生长,可能主要与其特异性下调survivin基因表达有关。特异性靶向survivin反义寡核苷酸可用于肺癌顺铂耐药的辅助治疗,其临床实际应用有待进一步研究。     

超声微泡介导VEGF反义寡核苷酸对裸鼠膀胱癌移植瘤生长的抑制作用

目的:探讨利用超声辐照微泡造影剂介导血管内皮生长因子(Vascular endotheliar growth factor,VEGF)反义寡核苷酸(Antisense oligodeoxynucleotides,ASODN)对人膀胱癌裸鼠移植瘤的抑制效应.方法:人膀胱癌裸鼠皮下移植瘤模型建立后,将成瘤后的裸鼠随机分为3组:UM+ASODN组每只裸鼠尾静脉注射微泡造影剂与VEGF-ASODN混合物,并对移植瘤体用超声波进行辐照;UM组每只裸鼠尾静脉注射微泡造影剂并以同样的条件进行体外超声辐照;单纯对照组每只裸鼠尾静脉注射生理盐水,各组处理每隔2d进行1次,共计5次.观察裸鼠移植瘤生长情况,免疫组化SP法测Ki67、CD34、VEGF的表达,计算微血管密度、细胞增殖指数,TUNEL法测细胞凋亡指数.结果:UM+ASODN组裸鼠皮下肿瘤生长速度较对照组明显减慢,肿瘤组织VEGF的表达及微血管密度较对照组降低(P<0.05);与对照组相比,UM+ASODN组肿瘤细胞的凋亡增加(P<0.01),而细胞增殖减少(P<0.05).结论:超声微泡介导的VECF反义寡核苷酸转染能有效抑制膀胱癌裸鼠皮下移植瘤的生长.     

c-myc反义寡核苷酸联合5-Fu对胃癌的体外及裸鼠体内抗肿瘤作用

目的:探讨c-myc反义寡核苷酸(ASODN)联合5-Fu对胃癌MKN-45细胞株的体内、外抗肿瘤作用.方法:采用脂质体介导c-myc反义寡核苷酸转染人胃癌细胞系,用四甲基偶氮唑盐法(MTT法)评价c-myc ASODN联合5-Fu对胃癌细胞增殖的影响;流式细胞仪(FCM)检测细胞凋亡率和细胞周期;RT-PCR及免疫组化方法检测c-myc基因表达变化;建立荷瘤小鼠模型,通过皮下瘤体内注射药物观察肿瘤体积和抑瘤率的变化.结果:c-myc反义寡核苷酸联合5-Fu可显著抑制细胞增殖,细胞凋亡率(23.4±1.9)%,较单用c-myc反义寡核苷酸(10.6±1.1)%和5-Fu(12.5±1.0)%的抑制作用更为显著(P＜0.01);RT-PCR及免疫组化显示ASODN和5-Fu均使c-myc mRNA和蛋白表达下降;体内实验显示c-myc反义寡核苷酸联合5-Fu可明显减少肿瘤体积,抑制肿瘤生长,抑瘤率达46.7%;较单一治疗组明显(P＜0.05).结论:c-myc基因反义寡核苷酸联合5-Fu能明显抑制胃癌MKN-45细胞增殖、诱导细胞凋亡和下调c-myc蛋白水平;且可有效抑制人胃癌裸鼠皮下肿瘤的生长.     

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.     

APPLICATION OF LORNOXICAM TO PATIENT-CONTROLLED ANALGESIA IN PATIENTS UNDERGOING ABDOMINAL SURGERIES

FOR anesthesiologis s ,treatingpostoperativepainhas alwaysbeen a problem.Althoughopioidshave been provedtobe effective,theirsideeffectscouldnotbeignored.With thedevelopmentofscienceand pharmacology,many drugs with aspectsof satisfactoryanalgesicefficacyand couldbe welltoleratedby patientshave been developed.And lornoxicamisone of them, which isa non-steroidalanti-inflammatorydrug (NSAID ), with analgesic, anti-infl-ammatory,andantipyreticproperties.Itseliminationhalf-time(3 to 5 hours) isle…     

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.     

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.