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。     

TSH的快速免疫检测在微流控芯片系统中的应用

目的建立一种可将免疫反应原理与化学发光技术相结合的微流控芯片系统。方法采用激光直接加工法制备微流控芯片,用双抗夹心法测定TSH。结果芯片系统可以在25min内完成TSH的检测流程,所需的标本和抗体试剂量为10μl,线性范围为0-50μIU/ml,批内CV平均为4.7%,批间CV平均为4.6%,平均回收率为95.3%。结论用于检测TSH的微流控芯片系统灵敏、快速、操作简便、可重复使用。     

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。     

CO2激光用于微流控免疫分析芯片加工的初步研究

目的建立微流控芯片的CO2激光直接微加工方法。方法构建激光加工装置，设置激光微加工程序，并对激光加工过程中一些影响因素进行分析，最后将加工后的芯片用于免疫分析。结果CO2激光可用于微流控芯片的加工，微流控芯片通道的深度主要受激光加工强度和激光加工次数的影响。对微通道表面的修饰可以改善通道的光滑度，采用该方法加工后的芯片可以用于免疫分析。结论CO2激光微加工法是一种操作方便、所需设备简单、加工周期短的微流控芯片加工方法。     

四种方法检测甲胎蛋白的比较

目的：比较时间分辨荧光免疫法（TRFIA）、微粒子化学发光法（CLA）、放射免疫法（RIA）和酶免疫法（EIA）四种方法检测甲胎蛋白（AFP）的线性、重复性、准确性和对比试验。方法：用不同浓度AFP分别进行了线性、精密度和回收试验。结果：TRFIA、CLA、RIA、和EIA四种方法的线性范围分别为1～1000ng／ml，1～1000ng／ml，10～400ng／ml和10—300ng；平均批内CV分别为3．75％、2．88％、5．08％和6．78％：平均批间CV分别为4．56％、3．45％、5．88％和7．72％；平均回收率分别为96．8％、97．1％、95．4％和92．6％。对比试验为TRFIA与CLA的相关系数r=0．991．TRFIA与RIA的相关系数r=0．963、TRFIA与EIA的相关系数r=0．843。结论：TR—FIA法和CLA法是目前检测AFP理想的定量方法。     

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。     

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。     

微流控芯片技术在生物学中的应用

分析装置的集成化、自动化及微型化现已成为物质分析和临床检测的研究热点。微流控芯片（microfluidic chip）作为一种新的微型分析平台建立于20世纪90年代，它通过微细加工技术在芯片上构建由储液池、微反应室、微管道等微功能元件构成的微流路系统，加载生物样品和反应液后，在压力泵或者电场作用下形成微流路，于芯片上进行一种或连续多种的反应，达到对样品的高通量快速分析的目的。微流控分析芯片由于具有高度集成性，可在一张芯片上完成采样、稀释、加试剂、反应、分离、检测等多种功能，     

基于微流控芯片的细胞迁移模型及其在黄芩苷药效学研究上的应用

基于微流控芯片技术建立一种体外细胞迁移模型,并将其用于黄芩苷药效的研究。设计与制作一种集成有微阀的微流控芯片;LIVE/DEAD染色试剂盒检测芯片中细胞活力;通过荧光素钠对芯片进行药物浓度梯度表征;DiI、DiO对细胞示踪染色观察微阀对细胞的隔离作用;每12小时显微镜拍照观察细胞的迁移状态并计算细胞迁移速率;ELISA法测定芯片细胞上清液中MMP-2、MMP-9和E-cadherin蛋白含量变化。实验结果表明,芯片中细胞活力接近100%;芯片可产生稳定的浓度梯度,且微阀对细胞有良好的隔离作用;黄芩苷对肝癌细胞HepG2的迁移具有显著抑制作用,并可显著降低MMP-2和MMP-9蛋白的分泌量。该研究为微流控芯片技术在中医药领域中的应用提供了一种新思路和新方法。     

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…     

Acupuncture Treatment of Vomiting

CASE HISTORY A female patient, 46 years old, head of the foreign affairs department of a certain university in Beijing, paid her first visit on October 9, 2006, with the chief complaint of vomiting for one month. She got vomiting after meals in early September. Before that, she had discomfortable sensation in the stomach due to angry with others, but she didn't pay much attention. Later, it developed into vomiting after eating. After the vomiting, the discomfort would be relieved, but with slight hypodynamia. She was once diagnosed as having 'neurogenic vomiting'. Having taken some western and Chinese drugs, the above symptoms were a little bit improved, but she would have nausea upon eating and with regurgitation. Because of the fear for vomiting, she did not dare to have food intake, with body weight reduction of 6 kilos in one month.     

Qiu Baoguo''''s Experience in Treating Consumptive Syndromes after Radio-Chemotherapies

Radiotherapy and chemotherapy are the important modern medical therapies for malignant tumors,yet they can also bring about serious local and systemic toxic side reactions so to decrease the patient;'s life quality,manifested by a series of consumptive symptoms.Having engaged in the combined work of Chinese and western medicine for nearly 50 years,the research fellow Qiu Baoguo in Henan Provincial Academy of TCM has developed his unique views on the TCM study of consumptive syndromes.The author of this essay had once the fortune tO follow Dr.Qiu in clinic,and specially would like to introduce in the following Dr.Qiu's experience in treating consumptive syndromes after radio-chemotherapies for patients with malignant tumor.     

Insomnia Due to Deficiency of Both the Heart and Spleen Treated by Acupuncture-Moxibustion and Chinese Tuina

OBJECTIVE: To observe therapeutic effects of the comprehensive therapy of acupuncture-moxibustion and Chinese Tuina for treatment of insomnia due to deficiency of both the heart and spleen. METHODS: 92 cases were divided randomly into the treatment group (treated by acupuncture-moxibustion and Chinese Tuina) and the control group (treated by acupuncture-moxibustion). RESULTS: The therapeutic effect of the treatment group was obviously superior to that of the control group (the CHI2 test showed P < 0.01). CONCLUSIONS: The comprehensive therapy of acupuncture-moxibustion and Chinese Tuina can give marked therapeutic effects for treatment of insomnia due to deficiency of both the heart and spleen.     

Analysis of CK-19,PDX-1,Nestin,Ngn3 in different donor islet purity in rats

Objective: To investigate if there are the CK-19, PDX-1, Nestin, Ngn3 positive cells in the donor islets of different purity in rats. Methods: Thirty male adult SD rats were randomly divided into 3 groups. Islets were isolated using digestion by ductal injection of collagenase. Group Ⅰ (n=10): Separating cell preparations were not purified, Group Ⅱ(n=10): Islet sediment was purified with 25% Ficoll400 ,Group Ⅲ (n=10): Islet sediment was purified with 25% and 11% Ficoll-400. The levels of protein of CK-19, PDX-1, Nestin and Ngn3 were detected by immunohistochemistry and the mRNA of CK-19, PDX-1, Nestin, Ngn3 was amplified by RT-PCR. Results: After two different purification methods applied, three islet preparations of different purities were obtained. The difference of islet purity was significant among various groups (P＜0.05). Compared with group Ⅱ and group Ⅲ,the protein and mRNA of CK-19, PDX-1, Nestin,Ngn3 were both higher in group Ⅰ; group Ⅲ was poorly expressed. Conclusions: The three different islet purity donor islet have different CK-19, PDX-1, Nestin, Ngn3 positive cells within them, indicating that there are some islet stem cells in the purified donor islet.     

Dr.Zhang Ren''''s Experience in the Acupuncture Treatment of Different Diseases with the Same Therapeutic Principle

Dr.Zhang Ren,the chief physician,is the chairman of Shanghai Acupuncture and Moxibustion Association.Having been engaged in medicine for about 40 years,he is experienced in treating various intractable diseases.In his long years of clinical practice,he advocates taking the TCM differentiation as the basis to seek for the acupuncture method for treatment of modern intractable diseases.The author of this essay had the fortune to follow Dr.Zhang in study.The following is a summary of Dr.Zhang's experience in the acupuncture treatment for different intractable diseases with the same therapeutic principle.     

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.     

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.