小细胞肺癌细胞系H446中肿瘤干细胞的分选与鉴定

 目的 明确能否以表面黏附分子CD133为干细胞标志物,使用免疫磁珠分选（magnetic cell separation,MACS）法分离小细胞肺癌细胞系H446细胞中的肿瘤干细胞。方法 以CD133为特异性分子标志物,使用MACS方法分选H446细胞,比较CD133阳性（CD133+）细胞与CD133阴性（CD133-）细胞在集落形成、自我更新、增殖分化、侵袭性、耐药性和成瘤性方面的不同。结果 CD133+细胞和CD133-细胞在集落形成能力、自我更新能力、增殖能力、分化能力、侵袭能力和耐药性方面基本相同。集落形成和成瘤性实验结果显示：CD133+或CD133-细胞中40%以上的细胞都能够形成含有100个细胞以上的大集落,增殖成为与未分选细胞相同的细胞群,并能在裸鼠身上成瘤。结论 表面黏附分子CD133不能作为分选小细胞肺癌H446细胞系中肿瘤干细胞的分子标志物,分选后的CD133+与CD133-细胞中含有相同的肿瘤干细胞。     

浅谈小细胞肺癌细胞系H446中肿瘤干细胞的分选与鉴定

目的:通过实验验证表面黏附分子CD133是否可以作为干细胞标志物。并通过这个试验过程浅谈小细胞肺癌细胞系H446中肿瘤干细胞的分选与鉴定。方法:在分选H446细胞(MACS方法)过程中得到CD133阳性(CD133+)细胞与CD133阴性(CD133-)细胞。对这两种细胞进行集落形成、自我更新、增殖分化、侵袭性、耐药性和成瘤性方面等项目的鉴定。结果:各种办法鉴定,发现两种细胞不存在差异。结论:分选小细胞肺癌H446细胞系中肿瘤干细胞方法中不能以表面黏附分子CD133为的标志物。     

胃癌干细胞标志物的研究进展

肿瘤干细胞(CSCs)具有类似成体组织干细胞的特性,可以自我更新并具有多向分化潜能。目前CSCs研究的难点主要是特异性标志物的筛选,CD44、CD24、上皮细胞黏附分子等胃癌干细胞表面标志物及侧群细胞功能性标志物的发现为胃癌干细胞的分离和纯化提供了特异性的分子标记,也为深入研究其与胃癌发生、发展、治疗、转移和预后提供了可行性依据。现就胃癌干细胞理论、表面标志物、功能性标志物及其标志物与胃癌的治疗、转移和预后的关系进行综述。     

CD90作为卵巢癌干细胞标志物的相关研究

目的 研究 CD90 作为分选卵巢癌干细胞标志物的可行性。方法 选择笔者实验室制备的人卵巢癌顺铂耐药细胞株 SKOV3-MD3,采用 CD90 单抗染色,应用流式细胞仪分选获得 CD90⁺ 细胞及 CD90-细胞。体外实验检测两个不同亚群细胞的体外悬浮球形成能力(自我更新能力)、分化潜能、表达干细胞标志物的差异,体内动物实验检测不同亚群细胞皮下成瘤能力是否有差异。结果 人卵巢癌细胞 SKOV3-MD3 中存在具有干性特征的 CD90⁺ 细胞,CD90⁺ 细胞比CD90- 细胞具有更高的体外悬浮球形成能力(自我更新能力)及分化潜能,表达更高水平的干细胞标志物,以及具有更强的皮下成瘤能力,差异有统计学意义。结论 CD90 可能是分选卵巢癌干细胞的分子学标志物。     

人肝癌细胞系Huh-7中CD133+细胞的分离及鉴定

目的 通过表面标志分选法富集人肝癌Huh-7细胞中CD133+细胞,并初步鉴定其特性.方法 采用流式细胞分选技术从人肝癌细胞系Huh-7中分选出CD133+细胞,并进行干细胞比例分析;通过对CD133+细胞体外成球能力及增殖能力检测,考察CD133+细胞的自我更新能力;观察CD133+细胞在非肥胖性糖尿病/重度联合免疫缺陷小鼠(NOD/SCID)体内的成瘤情况.结果 分选获得的CD133+细胞经无血清培养后阳性比例达90％以上;CD133+细胞体外无血清培养3d即可成球且生长速度较CD133-细胞快;CD133+细胞在NOD/SCID小鼠体内21 d左右即可形成异种移植瘤.结论 CD133+表面标志物分选方法可以高纯度富集CD133+细胞,利用CD133抗体分选获得的CD133+细胞具有肿瘤干细胞样特性.     

桑辛素对喉癌干细胞干性表型调控的作用研究

  目的  探索桑辛素对喉癌干细胞干性表型调控的影响。  方法  流式细胞仪分选并检测CD133+喉癌干细胞比例;肿瘤球形成实验检测CD133+喉癌干细胞的自我更新能力;Transwell实验检测CD133+喉癌干细胞的迁移能力;改良MTT实验检测化疗药物对CD133+喉癌干细胞的细胞毒性作用;免疫荧光染色、实时荧光定量PCR（RT-qPCR）及Western blot检测CD133+喉癌干细胞的干细胞标志物表达;在不同浓度的桑辛素处理CD133+喉癌干细胞后（以桑辛素0 μmol/L为对照）,通过肿瘤球形成实验、Transwell实验、改良MTT实验及Western blot,分别检测CD133+喉癌干细胞的自我更新能力、迁移能力、化疗药物的细胞毒性作用及干细胞标志物表达变化。  结果  流式细胞仪分选结果显示,CD133+喉癌干细胞占喉癌细胞的比例为（3.50±0.34）%;经过培养富集后,其比例可达（93.20±5.23）%。肿瘤球形成实验结果显示,与喉癌细胞相比,CD133+喉癌干细胞具有增强的自我更新能力 (P<0.001);Transwell实验显示,与喉癌细胞相比,CD133+喉癌干细胞的迁移能力增强（P<0.05）;改良MTT实验结果显示,与喉癌细胞相比,CD133+喉癌干细胞抵抗化疗药物（5-氟尿嘧啶及顺铂）的细胞毒性作用（P<0.05）;免疫荧光染色、RT-qPCR及Western blot结果显示,干细胞标志物（CD133、ALDH1、Sox2、ABCG2及N-cadherin）在CD133+喉癌干细胞中呈高表达水平。通过不同浓度的桑辛素处理CD133+喉癌干细胞,其自我更新能力降低（P<0.05）;迁移能力亦下降（P<0.05）;此外,桑辛素处理的CD133+喉癌干细胞对化疗药物的细胞毒性作用更加敏感（P<0.05）;Western blot结果显示,不同浓度的桑辛素处理的CD133+喉癌干细胞,其上述的干细胞标志物表达水平下调（P<0.05）。  结论  CD133+喉癌干细胞具有干性表型特征;桑辛素可减弱喉癌干细胞的干性表型,可能与下调其干细胞标志物表达相关。     

胃癌干细胞表面标志物的研究进展

胃癌干细胞（GCSC）在胃癌的发生、发展、侵袭、转移中发挥了重要的作用,因此探索灵敏度高、特异性强的GCSC表面标志物极其重要。本文阐述了常用及最新的GCSC标志物,其中CD44+ GCSC是最早发现具有启动肿瘤生长、维持肿瘤自我更新能力的独特亚群,是目前研究较为成熟的标志物之一。重复含G蛋白偶联受体5同样具有明确的细胞干性,但其作为GCSC标志物不是特异的,而CD133+细胞是否具有细胞干性,还需进一步探索其机制。SOX2、醛脱氢酶及其同工酶、NME/NM23核苷二磷酸激酶2具有干细胞潜能,为GCSC表面标志物提供了更多的可能及选择。     

片仔癀对肝癌干细胞增殖及凋亡的影响

目的利用克隆球培养法富集肝癌干细胞,研究片仔癀对肝癌干细胞增殖及凋亡的影响。方法采用克隆球培养法对肝癌干细胞进行富集,采用RT-PCR检测肝癌细胞及肝癌干细胞中自我更新基因Oct-4m RNA的表达;利用流式细胞仪检测肝癌细胞及肝癌干细胞表面标记蛋白CD133、CD90表达;采用台盼蓝计数方法检测片仔癀对肝癌干细胞增殖的影响;利用Hoechst33342染色法检测片仔癀对肝癌干细胞凋亡的影响。结果采用克隆球培养法可富集成球生长的肝癌干细胞;与肝癌细胞比较,肝癌干细胞Oct-4、CD133、CD90高表达（P<0.05）;片仔癀给药后可显著抑制肝癌干细胞的增殖（P<0.05）,并增加肝癌干细胞中凋亡的细胞。结论片仔癀具有抑制肝癌干细胞增殖及诱导其凋亡的作用。     

大鼠关节软骨干细胞的分离培养及其特性的鉴定

目的分离培养大鼠关节软骨干细胞(ACSCs)并鉴定其特性。方法运用纤连蛋白黏附法从大鼠关节软骨细胞中分选出ACSCs,流式细胞技术分别鉴定干细胞阳性表面抗原(阳性标志物CD90与CD44)和阴性表面抗原(阴性标志物CD45、CD31与CD34)在ACSCs中的表达水平,单克隆形成实验鉴定ACSCs的单克隆形成能力,成骨、成脂及成软骨诱导鉴定ACSCs的多向分化潜能。结果纤连蛋白可以特异性地分选出ACSCs。ACSCs高表达CD90与CD44,几乎不表达CD45、CD31与CD34。经过7d培养,单个ACSC可以形成大于32个细胞的单克隆细胞团。ACSCs具备很强的成骨、成脂和成软骨分化能力。结论大鼠关节软骨内存在ACSCs,ACSCs具有比较典型的干细胞特征。     

CD133的研究现状及其与神经胶质瘤的相关性

CD133(prominin-1)是5次跨膜(5-TM)糖蛋白家族中的第1个成员,这种蛋白局限性分布于胞质膜的突出处。人CD133分子存在2种亚型:CD133-1和CD133-2。作为肿瘤干细胞表面标志之一的CD133分子,它可以联合其他组织特异性、细胞特异性的分子进行肿瘤干细胞的筛选,其抗原位点可能会成为肿瘤治疗的靶位点。CD133为寻找神经胶质瘤干细胞特异标志物提供重要线索,是现在分离、纯化神经胶质瘤干细胞最重要的标志物,为肿瘤的发生、发展和预后的判断提供重要线索。但是,CD133作为神经胶质瘤干细胞特异性标志物的可靠性仍颇受争议。本文就近年来CD133的研究现状及其与神经胶质瘤相关性予以综述。     

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.     

Development on Adjustable Calibration Marker for Shock Wave Focus

Shock wave lithotripsy （SWL） is a treatment of choice for upper urinary stones. However, this procedure is inappropriate for obese patients because the focus is often unable to reach the target owing to the limited focal distance in shock wave source. Although treating such patients in a blast path may increase the application length of shock wave source, it＇s difficult to find this path on the lithotripter monitor. For this reason, we invented an adjustable calibration marker in order to set an effective focus in the shock wave hath.     

Study on the TCM Pathogenesis of Angiopathic Complications of Type II Diabetes

Excess production of reactive oxygen species(ROS)of mitochondrion mediated by hyperglycemia is the common pathogenesis of angiopathic complications of diabetes.TCM holds that the damp from the dysfunction of spleen.kidney and liver is the causative factor of complications of diabetes.This is similar to the mechanism of Ros resulting in angiopathic complications of diabetes.When the angiopathic complications of type II diabetes mellitus(T2DM)are difierentiated as caused by turbid damp in TCM can be explained as ROS.Since the obstruction of pathogenic damp in channels and collaterals is said to be the main pathogenesis,the treating principle should be dissolving the damp to remove the obstruction.     

Leptin, Ghrelin and TNF-α before and after Hypo-caloric Traditional Chinese Diet and Auricular Acupuncture

INTRODUCTION Obesity is a complex emergent problem, which can be possibly solved not only by the diet but also by the life style and promotion of a constant physical exercise. 1, 2 No doubt careful attentions must be given to the nutritional condition of obese people, the dietary habits, the somatic build (i.e. distribution of fat mass) and the organic functions linked to formation of the fat mass. All the parameters should be constantly monitored before, during and after a diet treatment. 3, 4, 5     

Relationship between Dysglycemia and Carotid Atherosclerosis in Tibetan Population

People with dysglycemia are at high risk for atherosclerotic diseases. This study aims at investigating the atherosclerotic vascular damage in dysglycemia and its metabolic origin in Tibetan population.     

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.     

CLINICAL SIGNIFICANCE OF SERUM CYTOKINES IL-1β,sIL-2R,IL-6,TNF-α,AND IFN-v IN ACUTE CORONARY SYNDROME

Objectives To explore serum cytokines levels (including IL-1 β, sIL-2R, IL-6, TNF-α, and IFN-v) and their significance in patients with acute coronary syndrome (ACS) and the subsequent follow-ups, with attempt to estimate the role of various serum inflammatory markers in the diagnosis and assessment of ACS.Methods The study population include 40 patients with acute myocardial infarction (AMI), 40 patients with unstable angina pectoris (UAP), and 40 controls. Among the 80 patients, 60 patients attended a follow up 4 months later. Serum inflammatory markers including IL-1 β, sIL-2R, IL-6, TNF-α, and IFN-v were measured by enzyme linked immunosorbent assay.Results Serum IL- 1 β, sIL-2R, IL-6, TNF-α were significantly higher in AMI group or UAP group compared to the control group and became significantly lower 4 months later in the follow-up patients. Serum levels of IFN-v shows no significant difference between AMI group or UAP group and controls, also showing no significant change when measured in follow up patients. There was no correlation between serum creatine kinase-MB isoenzyme levels and serum inflammatory markers either in UAP or AMI group. Furthermore, when divided into two subgroups using Wagner's QRS scoring system in the AMI group, there is no difference of each serum inflammatory marker between ≤ 6 scores group and ＞ 6 scores group.Conclusion Serum levels of certain inflammatory markers may have some diagnostic value for ACS, and can be a useful marker reflecting disease stability.