Internet网上人类基因组研究信息资源的利用

介绍INTERNET网上人类基因组信息资源的开发利用。     

利用生物信息学方法筛选一组胃癌相关EST特征序列

以生物信息学为研究方法，利用互联网基因组资源，对一组胃癌相关差异显示逆转录聚合酶链式反应方法获得的差异表达片段进行序列分析。方法：使用BLAST软件，将所得到的6条序列与EST数据库和基因组数据库进行比对分析，以期获得序列的特征信息，并筛选出有意义的EST序列。结果：显示有3条序列与非编码区长散布核元件LINE区域匹配；1条序列与CD9的外显子同源；1条为新的EST序列，已提交EST数据库并获得序列登陆号；1条序列尚须通过基因识别的方法进一步证实其特性。结论：通过生物信息学方法快速筛选获得胃癌相关已知EST序列1条，获得1条新EST序列，排除3条DNA污染序列。     

LRP16基因启动子的亚克隆及表达调控载体的构建

目的：为深入研究LRP16基因的表达调控机制，克隆及亚克隆了LRP16基因的启动子序列，构建LRP16基因启动子表达调控载体。方法：在NCBI的人类基因组数据库中截取并下载LRPl6基因转录起始位点5′侧翼区约3kb的基因组序列设计PCR扩增引物，从健康外周血中扩增获得该片段，以此序列为基础进行亚克隆，分别获得6条5’端不等、3’端平齐的片段，最后插入用于表达调控研究的pGL3—Basic载体。结果：获得了7条长度依次差别约为400bp的LRP16启动子克隆，分别构建了调控荧光素两报告基因的真核表达载体。结论：上述载体的成功构建为信息资源与实验手段的有效结合克隆启动子序列提出了一种模式，并为LRPl6表达克隆及启动子的活性分析奠定了基础。     

铜绿假单胞菌噬菌体PaP3整合位点的鉴定

目的：确定铜绿假单胞菌噬菌体PaP3在宿主菌基因组中的整合位点.方法：利用在噬菌体PaP3基因组中无酶切位点的Pst Ⅰ对溶原性细菌全基因组进行酶切,获得含有噬菌体PaP3全基因组及部分宿主菌DNA序列的大片段;然后进行第二次酶切,获得噬菌体基因组与宿主菌基因组的交界点序列,并进行克隆、测序,鉴定出杂合位点attR.根据λ噬菌体的整合机制,利用PCR反应扩增出另一杂合位点attL,根据测序结果及Blast检索确定attP与attB的位置和DNA序列.结果：经序列比对发现,attP及attB的核心位点分别位于噬菌体PaP3基因组中的t-RNAPro基因及铜绿假单胞菌PA3基因组中的t-RNALys基因中,其核心位点的DNA序列为21 bp（5＇-GGTCGTAGGT-TCGAATCCTAC-3.）.在核心位点的两侧存在正向重复序列及反向重复序列.结论：该研究为铜绿假单胞菌噬菌体的整合酶及整合机制的研究奠定了基础.     

豚鼠基因组26个多态性微卫星标记的筛选

目的：筛选豚鼠基因组的多态性微卫星标记，为豚鼠遗传质量控制及基因定位等工作奠定基础。方法：采用磁珠富集法和豚鼠基因组数据库筛选法获取微卫星位点序列，通过分析和初步筛选，挑选部分候选位点，根据其序列设计引物，对5种不同来源的豚鼠基因组DNA标本进行PCR扩增，以期获得多态性分子标记。结果：本实验采用磁珠法共获得微卫星序列304个，设计引物125对，最终获得多态性位点1个，暂未发现多态性的特异性位点17个；用数据库筛选法共获得微卫星序列292个，设计并合成相应引物178对，最终发现多态性位点25个，暂未发现多态性的特异性位点28个。结论：本实验共获得26个多态性微卫星标记，45个潜在的候选标记，为微卫星标记在豚鼠遗传质量监测及突变基因定位等工作的应用奠定了基础。     

人3p24-p25 478 kb完全基因组序列的注释

目的对我国承担的人类基因组测序项目中的3p24-p25478kb完全基因组序列进行注释。方法采用从头预测、数据库相似性比较和全长或部分mRNA序列与基因组序列的比对等手段,识别基因组序列中的编码蛋白质的基因,并采用EMBOSS软件包分析基因组序列的组分特征。结果识别出该区域中的两个编码蛋白质的已知基因,即SLC6A1和SLC6A11(其中后者在基因组草图序列中未被定位);对这段基因组序列中组分特征预测分析结果显示,该基因组序列的平均GC含量为47%,并存在3个假想的CpG岛,其中两个CpG岛分别位于130685～131516bp及307090～307870bp,另一个位于415585～416308bp。结论采用上述方法对基因组序列3p24-p25478kb进行了正确的注释,揭示了基因组序列中的有关的基因结构、GC含量、CpG岛等信息。     

中药三七根核糖体18SrRNA基因的序列分析

目的：研究中药三七根核糖体18SrRNA基因的序列特征。方法：根据模式植物拟南芥的18SrRNA的基因序列设计引物,对三七根的18SrRNA基因序列进行基因克隆、测序,并与模式植物拟南芥Arabidopsis thaliana以及人参属植物喜马拉雅人参Panax pseudoginseng Wall subsp.himalaicus var.angustifolius的相应序列进行比较。结果：通过对产于广西靖西的三七Panax pseudoginseng Wall.var.notoginseng(Bukill)Hoo et Tseng根的18SrRNA基因进行克隆测序,获得了三七根的18SrRNA基因的部分序列特征。结论：利用已完成全基因组序列测定的模式植物拟南芥的分子生物学信息来研究中药植物基源的基因组序列,将有助于加快中药植物基源的分子生物学研究进程。     

腹膜间皮瘤组织体细胞突变的分析

［摘要］目的：探讨临床标本全基因组测序中体细胞突变测定技术的应用。方法：对1例手术切除的腹膜间皮瘤肿瘤组织和其癌旁组织进行了第二代全基因组测序,并将结果与人类基因组数据库的参考序列进行比对。结果：发现639 717个肿瘤组织和癌旁组织细胞共有的单核苷酸变异（single nucleotide variation,SNV）,20 302个肿瘤组织独有的SNV和2 185个癌旁组织独有的SNV,还有223个位点,在肿瘤组织和癌旁组织以及人类基因组参考序列三者之间均不相同。结论：仅仅对个体2种组织样本和人类基因组参考序列比对,尚不能准确定位病变细胞的所有体细胞突变。因此,建立个体出生时的全基因组序列或预留出生时的DNA样本有助于排除正常组织体细胞突变的干扰。     

我国科学家首次提出“人类泛基因组”

本刊讯近日,我国科研人员在人类基因组研究中获得新的重大进展,发现人类基因组中存在着种群特异甚至个体独有的DNA序列和功能基因。科研人员还首次提出了＂人类泛基因组＂的概念。     

安徽省人乳头瘤病毒16型全基因组的结构特征

目的克隆安徽省人乳头状瘤病毒16型( HPV16)全基因组,分析HPV16全基因组序列并了解其结构特征。方法收集5例安徽省宫颈癌病理组织标本并提取总DNA,设计4对特异性引物分段克隆HPV16全基因组,测序后进行序列拼接及核苷酸序列分析。结果检测到1个宫颈癌病理样本中含有 HPV16并获得全基因组序列,序列全长7906 nts ( GenBank登录号：KC935953)。序列比对显示,安徽HPV16( HPV16-Anhui )与泰国HPV16和日本HPV16的全基因组核苷酸序列相似性最高,达99.5%。系统关系树分析显示,HPV16-Anhui与其他7个HPV16型聚成1个单独的分支。结论获得安徽省首例HPV16全基因组核苷酸序列,HPV16-Anhui与不同地区 HPV16亲缘关系均较近且基因组变异性较小。     

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.