以DCAP—P为底物的酸性磷酸酶连续监测法的建立

目的建立以2,6-二氯-4-乙酰基苯基磷酸盐(DCAP-P)为底物的一种新的酸性磷酸酶(ACP)的测定方法.方法在BTS-310半自动生化分析仪上研究连续监测法,建立实验参数.结果本法测定DCAP吸收峰为326nm;ACP的Km=0.139mmol/L;340nm时DCAP的摩尔消光系数为14440L/(mol×cm);缓冲液浓度0.1mol/L,最适pH5.4,底物浓度2.0mmol/L,延滞期60s,线性反应期15min;线性范围为0～1188u/L;血红蛋白(Hb)＜115.62mg/L对测定结果无影响;黄疸对测定结果无影响;本法总ACP与PAP(前列腺酸磷酶)的参考值范围分别为12.58～19.42u/L、2.52～9.04u/L.结论此方法重复性好,线性反应期长,线性范围广,干扰因素少,适用于自动生化分析.     

血清及高密度蛋白对氧磷酶活性的分光光度测定法

目的 在现有的实验条件下,获得稳定可靠的血清及高密度脂蛋白（high density lipoprotein,HDL) 中对氧磷酶J（paraoxonase,PON）活性的测定方法。方法 以乙酸苯酯为底物,在对氧磷酶的作用下水解,生成产物苯酚,测定5分钟内反应体系中在270nm处苯酚特征性的紫外吸收,换算为酶促反应速度。结果 通过观测底物浓度、pH、激活剂Ca^2 、抑制剂EDTA等对酶促反应的影响,得出最适的测定条件;底物浓度为5mmol/L,最适pH为8．0,必需激活剂Ca^2 浓度为2mmol/L。结论 在该反应条件下,测定不同浓度HDL或血清中该酶的活性,重复性和稳定性均符合要求。     

人白细胞弹性蛋白酶抑制剂体外筛选模型的建立及优化

目的探讨人白细胞弹性蛋白酶(HLE)与其底物反应的最适条件,建立一种高通量的HLE抑制剂体外筛选模型。方法通过测定反应产物———对硝基苯胺(p-NA)的光密度值,研究电解质浓度、反应温度、反应体系pH值、酶和底物作用浓度、反应时间及检测波长等对酶促反应的影响程度,确定酶与底物反应的最佳条件。结果确定的最优反应条件为:HLE(0.2 U/ml)30μl,底物(50 mmol/L)100μl,含1.0 mol/L NaCl的Tris缓冲液(pH 7.5),反应温度37℃,反应时间24 h,检测波长为545 nm。结论建立了一种稳定的HLE抑制剂体外筛选模型,该模型灵敏度高、准确性和重复性强,可用作筛选HLE抑制剂的平台。     

单胺氧化酶抑制药物体外筛选模型的建立

目的：构建一种能用于治疗神经退行性变疾病单胺氧化酶（monoamine oxidase,MAO）抑制药物的体外筛选模型。方法：采用紫外分光光度方法,以苄胺或5-羟色胺（5-hydroxytryptamine,5-HT）分别作为MAO-B或MAO-A的反应底物,测定其活性,并对MAO浓度、底物浓度、缓冲液浓度、pH值、反应时间和温度等进行优化。结果：研究确定的模型反应体系条件为：在温度37 ℃、酶预孵育时间20 min和反应时间60 min下,测定MAO-B活性最佳条件为100 mmol/L磷酸钾缓冲液（pH 7.6）,MAO和底物苄胺的终浓度分别为0.15 mg/ml和2.00 mmol/L;测定MAO-A活性最佳条件为100 mmol/L磷酸钾缓冲液（pH 7.4）,MAO和底物5-HT的终浓度分别为0.40 mg/ml和5.00 mmol/L。结论：建立了一种MAO抑制药物体外筛选的模型,该模型具有成本低、操作简便等特点,适合于大规模筛选MAO-B和MAO-A的抑制药物。     

液体双试剂测定血氨在全自动生化仪上的运用

目的在全自动生化仪上建立液体双试剂连续监测法测定血氨。方法用连续监测法对pH、谷氨酸脱氢酶浓度等反应条件进行实验研究,并进行方法学评价。结果双试剂连续监测方法,试剂Ⅰ:含三乙醇胺缓冲液(pH7.4)0.25mol/L,α-酮戊二酸15mol/L,NADH 0.3mmol/L;试剂Ⅱ:含谷氨酸脱氢酶≥160u/L。线性范围达321μmol/L,批内Cv=3.4%,批间Cv=8.7%。参考值范围(-x±2SD):10.5~44.1μmol/L。结论本方法操作简便,结果准确,方法合理,符合全自动条件,可推广使用。     

酶偶联法测定葡萄糖-6-磷酸脱氢酶方法学的建立

目的 建立葡萄糖-6-磷酸脱氢酶的酶偶联测定法.方法 用葡萄糖-6-磷酸作为底物启动酶促反应,于波长340nm连续监测NADPH生成速率.结果 酶的反应的最适pH 9.0,线性可达1000U/L,批内变异为3%～5%,回收率为98.9%～101.5%,与既往所述的方法具有良好的相关,r=0.99,Y=0.989X+0.035.结论 应用酶偶联法测定葡萄糖-6-磷酸脱氢酶能较好的排除干扰、结果准确,是一种值得广泛推广应用的方法.     

α-HBDH两种底物浓度活性测定的临床应用探讨

我们用13mmol/L和3.3mmol/L两种底物浓度用连续监测法测定α-HBDH的活性及与乳酸脱氢酶(LDH)活性的比值,测定温度30℃共测定体检合格的健康成人26例;心肌梗塞7     

血清及高密度脂蛋白中对氧磷酶活性的分光光度测定法

目的　在现有的实验条件下 ,获得稳定可靠的血清及高密度脂蛋白 (high density lipoprotein,HDL)中对氧磷酶 (paraoxonase,PON)活性的测定方法。方法　以乙酸苯酯为底物 ,在对氧磷酶的作用下水解 ,生成产物苯酚 ,测定 5分钟内反应体系中在 2 70 nm处苯酚特征性的紫外吸收 ,换算为酶促反应速度。结果　通过观测底物浓度、p H、激活剂 Ca2 + 、抑制剂 EDTA等对酶促反应的影响 ,得出最适的测定条件 :底物浓度为 5 m mol/ L ,最适 p H为 8.0 ,必需激活剂 Ca2 +浓度为 2 mmol/ L。结论　在该反应条件下 ,测定不同浓度 HDL 或血清中该酶的活性 ,重复性和稳定性均符合要求     

醛糖还原酶抑制剂筛选模型的建立

目的:建立醛糖还原酶抑制剂筛选模型,提供筛选醛糖还原酶抑制剂的方法。方法:从大鼠晶状体中提取醛糖还原酶,设定10 mmol/L DL-甘油醛为底物,0.15 mmol/L NADPH为辅酶等条件建立初始酶促反应体系,维持其它条件不变改变底物浓度、辅酶浓度、反应温度和反应时间,选择酶活性峰值点建立模型,通过比较槲皮素与芦丁对醛糖还原酶的抑制作用检验此模型。结果:醛糖还原酶酶促反应体系组成:总体积1 ml,各反应液的终浓度分别为:DL-甘油醛10 mmol/L,NADPH0.15 mmol/L,缓冲液PBS的pH=6.2,120 mmol/L,醛糖还原酶提取物100μl,反应温度37℃,反应时间3 min。IR50槲皮素相似文献   

一种诊断用谷氨酸脱氢酶酶学性质研究

目的研究一种国产谷氨酸脱氢酶的热稳定性、酸碱耐受性、最适温度、最适pH和动力学参数等性质。方法以α-酮戊二酸、铵离子及还原型辅酶I（NADH）为底物,在谷氨酸脱氢酶催化作用下,转变为谷氨酸和氧化型辅酶I（NAD）,利用NADH在340nm处有吸收峰,监测其反应速率（-△A Jmin）,计算谷氨酸脱氢酶的活力单位,采用Linewaeaver-Burk作图法（双倒数作图法）对谷氨酸脱氢酶的米氏常数进行测定。结果谷氨酸脱氢酶于50℃保温20min保留活力约为87．5％,而在60℃保温20min后残存活力为54％;最适反应温度为45℃;最适反应pH为7．5;在pH为5～10范围内酸碱耐受性较好,25℃放置20h残留活力为90％以上。结论动力学研究表明该酶与国外知名厂家产品相差不大。     

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.