微流控免疫芯片快速检测法的建立及其检测TSH和T4的效果

目的 在已制备的微流控芯片系统上建立基于顺磁微球的免疫检测方法，并将此方法用于检测促甲状腺激素（TSH）和甲状腺素（T4）。方法 采用数字激光刻蚀技术制备微流控芯片，以luminol-H2O2为发光体系，用双抗夹心法测定TSH，用免疫竞争法测定T4。结果 该检测系统可以在25min内完成检测，所需的标本和抗体试剂量为10山，TSH检测范围为0～50μIU／ml，批内CV平均为4．9％，批间CV平均为4．9％，平均回收率为93．74％；T4检测范围为9．375～300ng／ml，批内CV平均为4．9％，批间CV平均为4．7％，平均回收率为91．27％。结论 本方法检测TSH和T4具有操作简便、节约时间、节省试剂、稳定性好、准确度高的优点。     

微流控芯片用于甲胎蛋白化学发光免疫分析的研究

目的：建立一种可用于免疫分析的微流控芯片系统。方法：采用激光直接加工法制备微流控芯片，建立基于超顺磁珠的化学发光微流控免疫分析系统，用于甲胎蛋白（AFP）的测定。结果：芯片系统可在20min内完成AFP的分析，所需要的标本量和试剂量为5μl，线性范围为1～800ng／ml，批内变异系数（CV）平均为6，3％，批间CV平均为7．8％，回收率为94．3％。结论：微流控芯片系统是一种快速、耗费小、可重复使用的AFP测定方法。     

总T3快速检测微流控芯片的制作及应用研究

目的 建立一种可以用于免疫分析的微流控芯片系统。方法 采用激光直接加工法制备微流控芯片，建立由流体控制系统、免疫反应系统和信号处理系统构成的芯片系统，用免疫竞争法测定总T3。结果 芯片系统可以在20min内完成总T3的分析，所需要的标本量和试剂量为5μl。线性范围为1～10ng／ml，批内CV平均为7．2％。批间CV平均为10．7％，回收率为108．5％。结论 微流控芯片系统用于检测总T3具有快速、灵敏、稳定、可重复使用的特点。     

微流控芯片用于AFP快速检测的研究

目的建立一种可以用于免疫分析的微流控芯片系统。方法采用激光直接加工法制备微流控芯片,建立由流体拉制系统、免疫反应系统和信号处理系统构成的芯片系统,用于甲胎蛋白的测定。结果芯片系统可以在20min内完成AFP的分析,所需要的标本量和试荆量为5μl,线性范围为1-800ng／ml,批内CV平均为4．8％,批间CV平均为7．0％,回收率为96．3％。结论微流控芯片系统是一种快速、耗费小、可重复使用的AFP测定方法。     

毛细管电泳与微流控芯片电泳快速检测粪便中腺病毒的方法建立

目的 建立粪便中腺病毒的PCR-毛细管电泳-激光诱导荧光和PCR-微流控芯片电泳-激光诱导荧光快速检测方法。方法 采用试剂盒法提取粪便中腺病毒DNA,选择腺病毒六邻体基因保守区域进行PCR扩增反应。PCR产物分别用高灵敏度的SYBR Gold和SYBR Orange核酸荧光染料进行标记,在优化的毛细管电泳和微流控芯片电泳条件下分离,采用激光诱导荧光检测特异性PCR扩增产物。结果 腺病毒扩增产物在优化的毛细管电泳和微流控芯片电泳条件下可分别于9 min和6 min内完成检测。将PCR扩增产物测序,并与NCBI基因库中核酸序列比对,结果表明所选用的扩增序列特异性良好。商品化DNA Marker毛细管电泳和微流控芯片电泳对目标DNA片段bp值求得的日内精密度分别为1.14%～1.34%和1.18%～1.48%,日间精密度分别为1.27%～2.76%和2.85%～4.06%。毛细管电泳-激光诱导荧光法和微流控芯片电泳-激光诱导荧光法检测腺病毒的灵敏度分别为2.33×10² copies/mL和2.33×10³ copies/mL。两种方法用于实际粪便样品测定,准确度高。结论 本研究建立的PCR-毛细管电泳-激光诱导荧光和PCR-微流控芯片电泳-激光诱导荧光检测方法能够快速、灵敏、准确地检测出粪便中的腺病毒。     

一种基于微流控免疫磁珠快速检测登革热抗原的方法

目的建立一种灵敏、快速、方便的检测登革热抗原新方法,为登革热的临床诊断提供帮助。方法建立了一种基于微流控免疫磁珠快速检测登革热抗原检测方法。使用Solidwork软件设计微流控芯片,采用机械加工法和化学封接方式制备微流控芯片;利用化学偶联反应制备登革热抗体免疫磁珠;在微流控芯片载体上,以HRPTMB-H_2O_2为显色体系,利用双抗体夹心法原理检测登革热NS1抗原;最后,同时应用该方法与传统ELISA试剂盒对57例临床样品进行检测,分析该方法的正确度,并对两种方法的优缺点进行比较。结果微流控免疫磁珠方法整个检测过程仅需20 min,反应体系只需磁珠量10μg和样品量10μL;验证实验中,该方法可以将阴性和阳性明显区分开,阳性样本有显色,阴性、空白样本均无显色;性能上,该新型ELISA方法与传统ELISA方法结果阴阳性符合率达到100%。结论该新型ELISA检测方法具有简便、快速、节省试剂和所需样本少、灵敏度高、稳定性好、准确性高等优点,可以用于登革热抗原检测。     

微流控芯片电泳免疫法检测β_2微球蛋白

目的:建立一种基于微流控芯片电泳技术的快速微量蛋白检测方法。方法:用过量FITC-β2-MG抗体,与分析体系中β2-MG发生非竞争性免疫反应,FITC-β2-MG抗体和免疫反应形成的荧光免疫复合物通过微流控芯片电泳实现分离,激光诱导荧光检测荧光信号。结果:分别考察了电泳缓冲体系、pH值等对芯片电泳行为的影响,结果表明pH8.33、45mmol/LTBE缓冲液分离效果较好,在此基础上实现了β2-MG标准品的分析,建立了微流控芯片电泳免疫分析方法。结论:微流控芯片电泳免疫分析方法可在2min内完成一次样品的电泳分析,且试剂和样品消耗少,灵敏度高,改善了常规免疫分析的不足,显示出较好的应用前景。     

血清促甲状腺素免疫检测技术

血清促甲状腺素（TSH）是反映甲状腺功能最敏感的指标。血清TSH测定方法有多种,从最初的放射免疫分析法和免疫放射分析法到临床逐渐普及的免疫化学发光分析法、电化学发光免疫法和时间分辨荧光免疫法等,乃至近年出现的微流控芯片技术,血清TSH检测的灵敏度显著提高。文章就临床较为常用的血清TSH检测技术及存在的问题作一综述。     

血清促甲状腺素免疫检测技术

血清促甲状腺素(TSH)是反映甲状腺功能最敏感的指标.血清TSH测定方法 有多种,从最初的放射免疫分析法和免疫放射分析法到临床逐渐普及的免疫化学发光分析法、电化学发光免疫法和时间分辨荧光免疫法等,乃至近年出现的微流控芯片技术,血清TSH检测的灵敏度显著提高.文章就临床较为常用的血清TSH检测技术及存在的问题作一综述.     

RGD多肽功能化的微流控芯片识别/捕获肿瘤细胞的研究

目的 探讨经RGD多肽功能化后的微流控芯片系统识别/捕获肿瘤细胞的效果.方法 采用化学共价连接的方法固定RGD多肽,建立由流体控制系统、免疫反应系统构成的细胞识别芯片系统,用于细胞的筛选/捕获;选用不同密度的细胞进行进样试验,考察不同细胞密度对通道内细胞数量的影响,并讨论不同的孵育时间对细胞捕获的影响.结果 随着进样细胞密度的增加,进入通道内的细胞明显增多;此功能化后的芯片系统在15 min内对A549的捕获率为90%,在20min内可完全将细胞捕获于管壁.结论 经RGD多肽修饰的微流控芯片系统可在短时间内对A549细胞进行有效的筛选,且在细胞数量很低时,也能对其进行捕获.     

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.     

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…     

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.     

Acupuncture Combined with Spinal Tui Na for Treatment of Primary Dysmenorrhea in 30 Cases

Objective: To observe the therapeutic effects in acupunture treatment of primary dysmenorrhea combined with spinal Tui Na, and study its mechanism. Methods: Thirty cases of the treatment group were treated by acupuncture combined with spinal Tui Na, and thirty cases in the control group were treated by routine acupuncture. Results: The total effective rate was 93.3% in the treatment group, and 73.3% in the control group, with a significant difference between the two groups (P<0.05). Conclusions: Acupuncture combined with spinal Tui Na has good prospects for treatment of primary dysmenorrhea.     

Luo Lingjie's Experience in Treating Chronic Nephropathy

In treating chronic nephropathy,Luo Lingjie,a chief physician,pays attention to regulating the balance between yin and yang,treating infection if present,and removing pathogenic factors.He prescribes gentle drugs and uses carefully strongly warming-tonifying ones,emphasizes the importance of persuading the patient to persist in treatment with medication and nurse one's health for recuperation,and is good at combined use of TCM and western medicine therapy and brings the merits of various therapies into full play,with obvious theraoeutic effects.