肽核酸生物传感器检测系统构建时靶序列农度的影响及线性范围的确定

目的探索肽核酸生物传感器检测系统构建时靶序列浓度值对杂交效应的影响,以及靶序列浓度线性范围的确定.方法石英谐振式生物传感器阵列表面先固定上1.5μM的bis-PNA探针,观察终浓度为1pg/L至100μg/L的HBV DNA与探针进行杂交所引起的频率变化及反应所需时间.并对频率下降值与靶序列浓度做线性回归,确定其线性范围.结果靶序列浓度为1 pg/L时,传感器未能检测出任何频率的改变.随着浓度从10 pg/L增加到100μg/L,杂交反应引起的频率下降值呈先增加后趋于缓和的趋势,以10μg/L为分界线,而杂交平衡时间并没有显现出一定的变化趋势.在10pg/L到10μg/L的浓度范围内,浓度与频率下降值的线性回归方程为lgC=-2.745 5 0.069 1×△F,相关系数r=0.992 3.结论随着靶序列浓度的升高,杂交反应引起的频率下降值呈典型饱和曲线趋势,靶序列浓度的线性检测范围为:10 pg/L ～ 10μg/L.     

缓冲液pH值对肽核酸压电基因传感器阵列杂交效应的影响

目的 探索PBS缓冲液的pH值对肽核酸压电基因传感器阵列杂交效应的影响。方法 压电基因传感器阵列表面先固定上1．5μmol／L的bis-PNA探针，然后分别在pH5.8～8．0的PBS缓冲液中和相应的靶序列杂交，记录频率的下降值及反应所需的时间。结果 随着pH值从5.8升到8．0，杂交导致的频率下降值先高后低，以pH6．8为一个分水岭。杂交平衡时间在pH6.8之前总体上说相差尚不明显，但当pH值从6．8升到8．0时，却骤然增加。结论 bis-PNA和靶序列dsDNA的杂交时适用的pH范围很窄(pH6．6～7．0)。弱酸环境下(pH618时)最为容易，pH超过7．5就不适宜杂交。结果 从另一个侧面证实了在bis-PNA和dsDNA的杂交过程中“Hoogsteen链第一”的反应机制。     

探针浓度对压电传感器基因杂交效应的影响

目的 探讨探针浓度对压电传感器基因杂交效应的影响。方法 9种浓度的探针与同一浓度的靶序列在压电传感器阵列上进行杂交,计算机采集并分析传感器频率数据,比较各探针浓度下固定前后频率差,杂交达到平衡所需时间,杂交前后频率差,同时,根据较低浓度数据得出的杂交动力学参数。计算各浓度下的频率变化理论值。结果 随探针浓度增加,固定引起的频率变化值逐渐增加,杂交时间有缩短趋势,杂交前后的频率差值在探针浓度较低（1nmol/L-0.5μmol/L)时逐渐增大,而探;针浓度较高（1.0-3.5μmol/L)时反而减小,高浓度时杂交前后频率变化理论值与实际值存在明显差异。结论 在石英晶体传感器阵列上,基因杂交效率随探针浓度增加而增大,但探针浓度过高反而抑制杂交,其原因可能主要在于探针之间的空间构象关系和静电排斥。     

离子浓度对肽核酸压电基因传感器杂交效应的影响

目的　探索不同离子浓度的PBS缓冲液对肽核酸压电基因传感器阵列杂交效应的影响。方法　压电基因传感器阵列表面固定上 1 .5 μM的bis PNA探针后 ,分别观察在 1 0、2 0、5 0、1 0 0mM的PBS缓冲液环境中与相应的靶序列杂交 ,记录频率的下降值及反应所需的时间。结果　钠离子浓度从 1 0mM增加至 2 0mM时 ,杂交导致的频率下降值及平衡时间均增加 (P <0 .0 5 ) ;从 2 0mM增加至 5 0、1 0 0mM时 ,杂交导致的频率下降值各组间无显著差异 ,但杂交平衡时间却呈倍数增长。结论　离子浓度极大地影响了杂交反应的时间 ,低盐离子浓度更有利于提高bis PNA和dsDNA的杂交速率 ,高盐离子浓度则会大大降低bis PNA的结合速率。     

"RecA蛋白-互补单链DNA探针"生物信号放大系统用于压电基因传感器的研究

目的利用"RecA蛋白-互补单链DNA探针"与固定的肽核酸(bis-PNA)探针相结合,摸索最适的液相反应条件,以提高压电基因传感器基因传感表面的质量负载,从而提高传感器的灵敏度.方法基因传感器阵列固定上针对乙肝病毒的bis-PNA探针,加入靶DNA与之反应,然后加入预先处理过的不同浓度(0.5～6mg/ml)的"RecA蛋白-互补单链DNA探针".结果当RecA蛋白浓度为3mg/ml时,ATPγS浓度为12.5μM,所引起的频率变化最显著.结论在反应体系中加入"RecA蛋白-互补单链DNA探针",有效的提高了压电基因传感器的灵敏度.     

2种不同类型的核酸探针对压电基因传感器阵列检测特异性的影响

目的探讨用PNA作为探针与普通的DNA探针相比对压电传感器基因杂交的优越性.方法用生物素-亲和素法分别将PNA、DNA两种探针固定在基因传感器的金膜表面,分别与完全匹配、1个碱基错配、2个碱基错配的互补靶序列进行杂交,观察2种不同的探针与靶序列反应所引起的频率变化及所用的时间.结果相对于DNA探针,PNA探针识别碱基错配的能力明显高,且杂交时间更短.结论 PNA与靶序列DNA结合有高度的特异性,选用PNA作探针,可提高压电基因传感器检测的特异性.     

缓冲液离子浓度对新型PNA-LSAW基因传感器杂交效应的影响

目的 探讨PBS缓冲液的离子浓度对构建的肽核酸(PNA)漏声表面波(LSAw)基因传感器杂交效应的影响.方法 将1.0μmol/L的PNA探针固定在LSAW基因传感器表面,分别加入10、20、50、100mmol/L的PBS缓冲液,观察不同离子浓度的PBS缓冲液对PNA-LSAW基因传感器杂交效应的影响,记录分析传感器相位的变化值及反应所需的时间.结果 当钠离子浓度从10mmol/L增加至100mmol/L时,传感器的相位变化呈先升高后降低的趋势,以20mmol/L为分水岭.而杂交平衡时间增加显著.结论 离子浓度极大地影响了杂交反应的时间,低盐离子浓度更有利于提高PNA和靶序列的杂交速率,高盐离子浓度则会大大降低PNA的结合速率.离子浓度为20mmol/L时杂交最为适宜.     

缓冲液pH值对LSAW-bisPNA传感器检测系统的影响

目的 探讨PBS缓冲液pH值对LSAW-bisPNA基因传感器检测系统的影响.方法 LSAW-bisPNA基因传感器表面固定终浓度为1.0 μmol/L的bis-PNA探针,然后分别在pH 5.8～8.0的PBS缓冲液中和相应的靶序列杂交,记录相位变化及反应所需时间.结果 在不同pH值缓冲液中,bis-PNA与靶序列反应引起的相位变化有明显差异(P＜0.01),pH 6.6与pH 6.8时传感器响应信号最大,但两者反应所达到的平衡时间有明显差异(P＜0.05),pH值6.6时检测所需时间更短.结论 选择pH 6.6作为bis-PNA和靶序列dsDNA反应的最适pH值,在该pH值条件下传感器响应信号大,反应达平衡时间短.     

乙型脑炎病毒靶序列浓度对LSAW基因传感器检测系统的影响

目的 探讨乙型脑炎病毒(Japanese encephalitis vires,JEV)靶序列浓度值对漏声表面波(LSAW)基因传感器检测系统的影响.方法 LSAW基因传感器表面固定终浓度为0.5 μmol/L的JEV探针,然后用pH 7.6的10 mmol/L PBS缓冲液(含Na+ 0.3 mol/L)中和终浓度为0.05、0.1、0.5、1.0、1.5、2.0 μmoL/L的JEV靶序列杂交,分别记录相位变化及反应所需时间.结果 与JEV探针反应引起的相位变化是先缓慢上升后趋于缓和的趋势,以1.0 μmol/L靶序列浓度为分界线;杂交平衡时间上除0.05 μmol/L的时间短以外,其他的未见明显变化趋势,当靶序列浓度为1.0 μmol/L,杂交反应平衡时间为33 min.结论 随着靶序列浓度的升高,杂交反应引起的相位变化呈典型的饱和曲线趋势;靶序列浓度为1.0 μmol/L、杂交反应的平衡时间为33 min,是LSAW基因传感器杂交的最适反应条件.     

离子强度对压电传感器基因杂交动力学的影响

目的：探讨盐离子浓度对压电传感器基因杂交动力学的影响。方法：设计并合成金黄色葡萄球菌耐药基因探针及与其完全互补的靶序列，在石英谐振压电传感器阵列上进行杂交，杂交缓冲液中含5种NaCl浓度，计算机采集并分析传感器频率数据得出杂交动力学参数，同时比较各盐离子浓度下杂交达到平衡所需时间，杂交前后的频率差，平衡后频率波动幅度大小。结果：随NaCl浓度增加，结合速率常数，杂交平衡常数，最大杂交量均呈现明显增大趋势。而离解速率常数呈下降趋势，杂交前与杂交平衡后的频率差值增大，同时，杂交平衡后频率波动幅度相应增大，杂交达平衡所需时间延长。结论：一定的盐离子浓度促进基因杂交，但盐离子浓度过高则对晶体振荡性能产生负面影响。     

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…     

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.     

猪肺磷脂注射液联合经鼻持续气道正压通气对呼吸衰竭早产儿的临床疗效及肌酸激酶同工酶活性的影响

目的 探讨猪肺磷脂注射液联合经鼻持续气道正压通气(NCPAP)对呼吸衰竭早产儿的临床疗效及肌酸激酶同工酶活性(CK-MB)的影响.方法 选取呼吸衰竭早产儿80例,分为观察组和对照组各40例.对照组采用NCPAP给氧治疗,观察组给予NCPAP给氧联合猪肺磷脂气管内给药.观察两组患儿治疗前及治疗12h、24 h后PaO2、PaCO2、血氧饱和度(SaO2)、pH的变化情况,检测治疗前及治疗5d后血清CK-MB水平;评估两组患儿的临床治疗效果.结果 两组患儿PaO2、PaCO2、SaO2、pH比较,差异均有统计学意义(P＜0.05),其中观察组治疗后的PaO2、SaO2、pH均高于对照组,PaCO2则低于对照组.两组的PaO2、SaO2、pH均随观察时间延长而升高(P＜0.05),PaCO2均随观察时间的延长而降低(P＜0.05).观察组治疗有效率为87.5％,显著高于对照组的70.0％ (P ＜0.05).治疗5d后两组患儿血清CK-MB水平均较前降低(P＜0.05),且观察组明显低于对照组(P＜0.05).结论 猪肺磷脂注射液气管内给药联合NCPAP可以显著降低呼吸衰竭早产儿CK-MB的含量,提高治疗有效率,起到很好的呼吸循环支持作用.     

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.     

Survey and practice of reporting quality of randomized controlled clinical trials on traditional Chinese medicine

Evidence obtained from randomized controlled trials （RCTs） has been generally accepted as the gold standard in the evaluation of clinical effectiveness. Readers need to understand the trial design, implementation, results, analysis and interpretation, so as to fully Jnderstand the results of RCTs. Thus, the investigators of RCTs have to report these items in a complete, accurate and clear manner. Since 1998, we have conducted several evaluations on the reporting quality of RCTs published in Chinese journals on traditional Chinese medicine （TCM） and results have shown that there is an urgent need for higher quality RCTs on TCM.