小鼠RAPD-PCR反应体系及扩增程序的筛选

目的:筛选优化小鼠RAPD-PCR反应体系及扩增程序.方法:以40条引物对9个品系的小鼠基因组DNA在各种不同条件下进行RAPD-PCR,分析比较扩增结果.结果:在下列反应体系及扩增程序下结果较好:在25 μl的反应体系中含有10×buffer 2.5 μl,dNTP 200μmol/L, Mg2+ 2.5 mmol/L,引物0.2 μmol/L,Taq酶1.5 U,模板DNA 20 ng,扩增程序为94℃变性1 min,38℃退火1 min,72℃延伸2 min,40个循环后接10 min延伸.结论:以上述反应体系及扩增程序进行小鼠RAPD-PCR,扩增结果稳定.     

药用百合DNA提取和RAPD条件的优化

目的确定药用百合最佳随机扩增多态DNA(RAPD)分析方法。方法以药用百合新鲜鳞叶为材料,研究DNA的提取方法,并对影响RAPD反应的各因素进行优化。结果药用百合RAPD反应体系及程序为:在40μL反应体系中,含40 ng模板DNA,0.3μmol/L随机引物,0.2 mmol/L dNTPs,1.7 mmol/L Mg2 ,3μL 10×PCRBuffer,1.5 U Taq酶;扩增程序为:第一步94℃预变性2 min,第二步94℃变性1 min,第三步36℃退火45 s,第四步72℃延伸90 s,返回至第二步重复40个循环,第六步72℃延伸10 min,最后4℃保存以备电泳。结论建立了药用百合RAPD的优化反应体系及程序。     

人源细粒棘球蚴的RAPD分析体系建立

目的建立PCR—RAPD反应体系，提升细粒棘球蚴随机扩增多态性DNA（RAPD）遗传多态性研究的稳定性，为研究细粒棘球蚴的分类学以及包虫病的分子流行病学奠定基础。方法采用改进的酚-氯仿抽提法提取细粒棘球蚴基因组DNA，用单因素筛选方法建立PCR—RAPD分析体系。结果采用改进的酚／氯仿抽提方法提取细粒棘球蚴基因组DNA质量高，适用于PCR—RAPD扩增分析；在25μL体系中，RAPD—PCR扩增反应各成分的最佳浓度为：dNTP0．2—0．3mmol／L、Taq酶2．5U、随机引物0．8—1．4μmol／L，模板为60ng。PCR扩增最佳条件为：95℃预变性5min；94℃变性30s、32℃退火45s、72℃延伸1min、共35个循环；最后72℃延伸10min；初步从20条引物中筛选出11条带型丰富的随机引物。结论酚-氯仿适用于提取细粒棘球蚴基因组DNA，优化的RAPD反应体系和筛选出的随机引物适合进行细粒棘球蚴RAPD遗传多态性分析。     

凋亡相关基因bcl-2和Bax在大鼠应激性溃疡中的表达及作用

目的筛选优化小鼠RAPD-PCR反应体系及扩增程序。方法以2条引物对4个品系的小鼠基因组DNA在各种不同条件下进行RAPD-PCR,分析比较扩增结果。结果扩增结果较好的反应体系及扩增程序如下:在25 ml的反应体系中含有10×buffer2．5μl,dNTP 200 mmol／L,Mg2+ 2．5mmol／L,引物0．2 mmol/L,Taq酶1．25 U,扩增程序为94℃变性1 min,40℃退火1 min,72℃延伸90 s,40个循环后72℃延伸5 min。结论这一反应体系和扩增程序在我们实验室得到了较好的扩增效果。     

两面针ISSR-PCR反应体系的建立及优化

【目的】建立适合两面针简单重复序列—聚合酶链反应(ISSR-PCR)体系和扩增程序。【方法】采用改良的高盐低pH法提取两面针基因组DNA,通过单因素试验和正交试验相结合对ISSR反应体系中的主要成分(Mg2+、Taq DNA聚合酶、引物和模板)进行筛选和优化,并考察退火温度对扩增结果的影响。【结果】最佳的PCR反应体系为:总体积为25μL,含1.50 mmol/L Mg2+、150μmol/L dNTPs、0.04 U/μL Taq DNA聚合酶、0.60μmol/L引物、2.40 ng/μL DNA模板。扩增程序为:94℃预变性5 min;94℃变性30 s,51.4℃退火45 s,72℃延伸2 min;35个循环;72℃再延伸10 min。【结论】优化了两面针ISSR-PCR反应体系和扩增程序,为两面针ISSR遗传多样性分析打下基础。     

白芷ISSR-PCR反应体系的建立和优化

目的:建立白芷的ISSR最佳反应体系及PCR扩增参数,为今后利用ISSR标记技术进行白芷鉴定及种质遗传多样性分析提供一个标准化程序.方法:以白芷基因组DNA为模板,分析了ISSR反应体系中模板DNA、Mg2 、dNTPs、引物浓度,TaqDNA聚合酶用量及退火温度对ISSR-PCR扩增的影响.结果:建立了重复性好,分辨率高的ISSR反应体系,即在25μL反应体系中,模板DNA浓度为10～30 ng,2.5μ10×PCR buffer(Mg2 free),MgCl2 2.0 mmol/L,dNTPs 0.2 mmol/L,Taq酶1.0U,引物0.6 pμmol/L;扩增程序为:94℃预变性5 min;94℃变性30 s,引物在筛选后的最佳退火温度退火45 s,72℃延伸2min,共计39个循环,循环结束后在72℃延伸5 min.结论:建立了适用于白芷的ISSR反应体系.为今后利用ISSR标记技术开展白芷遗传变异分析和构建遗传图谱奠定了基础.     

黄花蒿ISSR-PCR反应体系的优化

目的:建立黄花蒿(Artemisia annuaL.)的ISSR最佳反应体系及PCR扩增参数,为今后利用ISSR标记技术开展黄花蒿种间遗传多样性分析提供一个标准化程序。方法:以黄花蒿DNA为材料,分析了ISSR反应体系中的DNA、Mg2 、dNTPs浓度,TaqDNA聚合酶用量以及退火温度对ISSR-PCR扩增的影响。结果:建立了重复性好,分辨率高的ISSR反应体系,即在25μL反应体系中,模板DNA浓度为15~45 ng,2.5μL的10倍Buffer(Mg2 free)缓冲液,MgCl22.0 mmol/L,dNTPs为0.15 mmol/L,Taq酶1.0 U,引物1.0μmol/L;扩增程序为:94℃预变性5 min;94℃变性30 s,引物在筛选后的最佳退火温度退火45 s,72℃延伸2 min,共计40个循环,循环结束后在72℃延伸5 min。结论:建立了适用于黄花蒿的ISSR反应体系,为今后利用ISSR标记技术开展黄花蒿种间遗传变异分析和构建遗传图谱奠定了基础。     

香加皮ISSR—PCR条件体系的优化

采用正交设计法、均匀设计法,对影响香加皮IssR—PCR43体系的引物、Mg2＋、dNTP、TaqDNA聚合酶浓度进行优化,通过两种方法的比较建立了适合香加皮ISSR-PCR的反应体系：在20μL反应体系中,含Mg2＋1mmol／L、dNTP0．15mmol／L、引物1．0μmol／L、TaqDNA聚合酶1U、buffer2．5μL和模板DNA约50ng。在此基础上对扩增程序中的退火温度进行筛选,通过比较将退火温度定为50℃,最终扩增程序定为：94℃预变性4min接着进行40个循环：94℃变性30S,50~C退火1min,72℃2延伸90s;循环结束后72℃延伸10min。     

黄花蒿SRAP-PCR反应体系的建立与优化

目的用序列相关扩增多态性(SRAP)这一新的分子标记技术建立稳定可重复的黄花蒿Artemisia annua的SRAP最佳反应体系。方法以黄花蒿DNA为模板,分析了SRAP反应体系中的一些重要参数对PCR扩增的影响。结果建立了稳定可重复的SRAP反应体系;25μL反应体系中,模板DNA为30～105ng,MgCl2为2.0mmol/L,dNTP为0.25mmol/L,Taq酶为1.0U,引物为2.0μmol/L;扩增程序:94℃预变性3min,94℃变性1min,35℃复性1min,72℃延伸1min,5个循环;94℃变性1min,50℃复性1min,72℃延伸1min,35个循环,72℃延伸7min。结论本研究建立的黄花蒿SRAP体系重复性好,稳定性强。     

红景天属植物DALP反应体系的建立

目的 利用直接扩增片段长度多态(DALP)这一新分子标记技术建立一套稳定的红景天属植物的DALP反应体系。方法 以红景天基因组DNA为模板,对DALP反应程序的一些重要参数进行摸索和优化试验。结果 建立了一套稳定性强、可靠性高的DALP应用反应体系;经过大量重复性实验,反应体系为20μL,Mg2 浓度为2.5 mmol/L,dNTPs浓度为1.25 mmol/L,模板DNA的量为60 ng,5 pmol/L选择性引物1μL,5 pmol/L反向引物3 μL,引物浓度比为1:3,Taq酶2 U。反应过程为:95℃预变性5 min、94℃变性30 s、50℃退火30 s、72℃延伸1 min;30个循环,再72℃延伸10 min,完成整个PCR反应。结论 该体系可有效地应用于红景天属药材的分类鉴定和地道性鉴定。     

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.     

A preliminary screening study on the associated proteins in human psoriasis vulgaris by serum proteomics technologies

Objective：To investigate the optimum screening conditions of associated proteins in human psoriasis vulgaris by serum proteomics technique, and to screen the different expression proteins related with psoriasis vulgaris. Methods：Serum samples of peripheral blood were collected from newly diagnosed psoriasis vulgaris patients in the clinic, and 20 matched healthy persons. Serum albumin IgG was removed by filtering with ProteoExtract Albumin/IgG. After comparative proteomics analysis the different protein spots were identified using 2-DE and MS. Results ：Electrophoresis figures with high resolution and reproducibility were obtained. Three different expression proteins were found only in the serum from psoriasis vulgaris patients, while nine other different proteins expressing from healthy volunteers. Conclusion：The protein expression was different in the serum between the psoriasis vulgaris patients and healthy volunteers. It was hoped that we could find the biomarkers related to psoriasis vulgaris by using proteomics.     

A CASE OF HUGE GASTRIC DISOPYROBEZOAR

PYTOBEZOARS are common bezoars in gastrontestinal tract, commonly seen in stomach and mall intestine,1but huge disopyrobezoars are     

Acupuncture Treatment of Chest Bi Syndrome

CASE HISTORY A male,a retired officer of 61 years old,paid his first visit on May 12,2006.The patient stated that 2 weeks before he suddenly got paroxysmal precordial choking pain,with irregular attacks.Each attack would last several seconds to several minutes.The attack was accompanied with heavy oppressed sensation in the precordial region,palpitation,lassitude,and in severe cases,sweating,which was once diagnosed by a hospital as'coronary heart disease'.Recalling the case history,the patient said that several days before he got angry with others because of some affairs in work,and he was not so used to the life pattern since he had just retired from his busy working post.At usual times,he was quick tempered and peevish.In recent days,he had poor appetite,and loose stool.     

Instructions to Contributors

The English edition of Chinese Journal of Integrative Medicine is edited and published by the Chinese Journal of Integrated Traditional and Western Medicine Press.Original scientific papers of advanced clinical and experimental medicine in the field of complementary/alternative medicine or integrative medicine are expected.