通过大量培养克隆有 HPV11、16、18、51型 DNA 序列的菌株,扩增其质粒,并采用碱变性法提取、纯化质粒 DNA,然后对质粒进行酶切、电泳回收 HPV DNA 片段并进行生物素标记,制备出生物素标记的 HPV11、16、18、51型 DNA 探针.用斑点杂交方...通过大量培养克隆有 HPV11、16、18、51型 DNA 序列的菌株,扩增其质粒,并采用碱变性法提取、纯化质粒 DNA,然后对质粒进行酶切、电泳回收 HPV DNA 片段并进行生物素标记,制备出生物素标记的 HPV11、16、18、51型 DNA 探针.用斑点杂交方法检测探针的灵敏度为 HPV11、16型0.05pg,HPV18 0.25 pg,HPV51 0.1 pg.展开更多
To screen species-specific DNA probes for identification of Fallopia muhiflora, the genomic DNA （gDNA） suppression subtraction hybridization （SSH） between F. muhiflora and F. muhiflora var. ciliinervis was firstly...To screen species-specific DNA probes for identification of Fallopia muhiflora, the genomic DNA （gDNA） suppression subtraction hybridization （SSH） between F. muhiflora and F. muhiflora var. ciliinervis was firstly performed. The obtained differential gDNA fragments by SSH were then hybridized with gDNA ar- rays consisting of multiple whole genomes of several species （adulterants and/or closely related species of F. muhiflora） and four differential fragments were screened uniquely representing F. muhiflora, which could be used as F. muhiflora species-specific probes. The screened DNA probes were tested by reverse dot blot hybridization and the results demonstrated that these probes could be used reliably to identify F, muhiflora. The species-specific DNA probes obtained in this study exhibited broad application prospects in the preparation of gene chips for identifying Chinese traditional medicines and the authentication of germplasm re- sources and crude drugs of F. muhiflora.展开更多
Cancer-related diseases represent the second overall cause of death worldwide.Human papilloma virus(HPV)is an infectious agent which is mainly sexually transmitted and may lead to HPV-associated cancers in both men an...Cancer-related diseases represent the second overall cause of death worldwide.Human papilloma virus(HPV)is an infectious agent which is mainly sexually transmitted and may lead to HPV-associated cancers in both men and women.Almost all cervical cancers are HPV-associated,however,an increasing number of head and neck cancers(HNCs),especially oropharyngeal cancer,can be linked to HPV infection.Moreover,anogenital cancers,including vaginal,vulvar,penial,and anal cancers,represent a subset of HPVrelated cancers.Whereas testing and prevention of cervical cancer have significantly improved over past decades,anogenital cancers remain more difficult to confirm.Current clinical trials including patients with HPV-related cancers focus on finding proper testing for all HPV-associated cancers as well as improve the currently applied treatments.The HPV viral oncoproteins,E6 and E7,lead to degradation of,respectively,p53 and pRb resulting in entering the S phase without G1 arrest.These high-risk HPV viral oncogenes alter numerous cellular processes,including DNA repair,angiogenesis,and/or apoptosis,which eventually result in carcinogenesis.Additionally,a comprehensive analysis of gene expression and alteration among a panel of DNA double strand breaks(DSB)repair genes in HPV-negative and HPV-positive HNC cancers reveals differences pointing to HPV-dependent modifications of DNA repair processes in these cancers.In this review,we discuss the current knowledge regarding HPV-related cancers,current screening,and treatment options as well as DNA damage response-related biological aspects of the HPV infection and clinical trials.展开更多
The serum samples and corresponding cervical swabs were collected from 50 women with genital warts from Tianjin city, China. The neutralizing antibodies against HPV-16, -18, -58, -45, -6 and -11 in serum samples were ...The serum samples and corresponding cervical swabs were collected from 50 women with genital warts from Tianjin city, China. The neutralizing antibodies against HPV-16, -18, -58, -45, -6 and -11 in serum samples were tested by using pseudovirus-based neutralization assays and HPV DNAs in cervical swabs were also tested by using a typing kit that can detect 21 types of HPV. The results revealed that 36% （18/50） of sera were positive for type-specific neutralizing antibodies with a titer range of 160-2560, of which 22%（11/50）, 12%（6/50）, 10%（5/50）, 4%（2/50）, 4%（2/50） and 2%（1/50） were against HPVs -6, -16, -18, -58, -45 and -1 l, respectively. Additionally, 60% （30/50） of samples were HPV DNA-positive, in which the most common types detected were HPV-68（18%）, HPV-16（14%）, HPV-58（12%）, HPV-33（8%） and HPV-6, HPV-11, HPV-18 and HPV-52 （6% each）. The concordance between HPV DNA and corresponding neutralizing antibodies was 56% （28/50） with a significant difference （P〈0.05）. The full-length sequences of five HPV types （HPV -42, -52, -53, -58 and -68） were determined and exhibited 98%-100% identities with their reported genomes. The present data may have utility for investigating the natural history of HPV infection and promote the development of HPV vaccines.展开更多
基金Supported by Fund of Guangdong Provincial Administration of Traditional Chinese Medicine(20111251)
文摘To screen species-specific DNA probes for identification of Fallopia muhiflora, the genomic DNA （gDNA） suppression subtraction hybridization （SSH） between F. muhiflora and F. muhiflora var. ciliinervis was firstly performed. The obtained differential gDNA fragments by SSH were then hybridized with gDNA ar- rays consisting of multiple whole genomes of several species （adulterants and/or closely related species of F. muhiflora） and four differential fragments were screened uniquely representing F. muhiflora, which could be used as F. muhiflora species-specific probes. The screened DNA probes were tested by reverse dot blot hybridization and the results demonstrated that these probes could be used reliably to identify F, muhiflora. The species-specific DNA probes obtained in this study exhibited broad application prospects in the preparation of gene chips for identifying Chinese traditional medicines and the authentication of germplasm re- sources and crude drugs of F. muhiflora.
基金supported by the Pamela and Wayne Garrison Distinguished Chair in Cancer Research to J.C.J.C.also received support from the CPRIT awards(Grant Nos RP160667 and RP180813)NIH grants(Grant Nos P01CA193124,R01CA210929,R01CA216911,and R01CA216437)。
文摘Cancer-related diseases represent the second overall cause of death worldwide.Human papilloma virus(HPV)is an infectious agent which is mainly sexually transmitted and may lead to HPV-associated cancers in both men and women.Almost all cervical cancers are HPV-associated,however,an increasing number of head and neck cancers(HNCs),especially oropharyngeal cancer,can be linked to HPV infection.Moreover,anogenital cancers,including vaginal,vulvar,penial,and anal cancers,represent a subset of HPVrelated cancers.Whereas testing and prevention of cervical cancer have significantly improved over past decades,anogenital cancers remain more difficult to confirm.Current clinical trials including patients with HPV-related cancers focus on finding proper testing for all HPV-associated cancers as well as improve the currently applied treatments.The HPV viral oncoproteins,E6 and E7,lead to degradation of,respectively,p53 and pRb resulting in entering the S phase without G1 arrest.These high-risk HPV viral oncogenes alter numerous cellular processes,including DNA repair,angiogenesis,and/or apoptosis,which eventually result in carcinogenesis.Additionally,a comprehensive analysis of gene expression and alteration among a panel of DNA double strand breaks(DSB)repair genes in HPV-negative and HPV-positive HNC cancers reveals differences pointing to HPV-dependent modifications of DNA repair processes in these cancers.In this review,we discuss the current knowledge regarding HPV-related cancers,current screening,and treatment options as well as DNA damage response-related biological aspects of the HPV infection and clinical trials.
文摘The serum samples and corresponding cervical swabs were collected from 50 women with genital warts from Tianjin city, China. The neutralizing antibodies against HPV-16, -18, -58, -45, -6 and -11 in serum samples were tested by using pseudovirus-based neutralization assays and HPV DNAs in cervical swabs were also tested by using a typing kit that can detect 21 types of HPV. The results revealed that 36% （18/50） of sera were positive for type-specific neutralizing antibodies with a titer range of 160-2560, of which 22%（11/50）, 12%（6/50）, 10%（5/50）, 4%（2/50）, 4%（2/50） and 2%（1/50） were against HPVs -6, -16, -18, -58, -45 and -1 l, respectively. Additionally, 60% （30/50） of samples were HPV DNA-positive, in which the most common types detected were HPV-68（18%）, HPV-16（14%）, HPV-58（12%）, HPV-33（8%） and HPV-6, HPV-11, HPV-18 and HPV-52 （6% each）. The concordance between HPV DNA and corresponding neutralizing antibodies was 56% （28/50） with a significant difference （P〈0.05）. The full-length sequences of five HPV types （HPV -42, -52, -53, -58 and -68） were determined and exhibited 98%-100% identities with their reported genomes. The present data may have utility for investigating the natural history of HPV infection and promote the development of HPV vaccines.
