The impact of habitat fragmentation and isola-tion on the genetic diversity of populations has attracted much attention in studies of meta-population and conserva-tion biology. In this work, using the randomly amplifi...The impact of habitat fragmentation and isola-tion on the genetic diversity of populations has attracted much attention in studies of meta-population and conserva-tion biology. In this work, using the randomly amplified po-lymorphic DNA (RAPD) technique, we studied the genetic diversity of central, peripheral and peninsular populations of ratlike hamster, which were collected in five locations of the North China Plain and its surrounding areas, in 1999. The study revealed that, i ) the genetic diversity of central population of Raoyang County 】 the sub-central populations of Gu’an County and Taikang County 】 the peripheral population of Shunyi District 】 the peninsular population of Mentougou District; ii) the genetic diversities of the five populations were positively correlated to the nearest dis-tances to the peripheral line of population distribution; iii) there were significant differences of gene frequencies of some RAPD fragments among the five populations. More RAPD fragments disappeared in展开更多
AIM: To clone the cDNA of UGT1A9 from a Chinese human liver and establish the Chinese hamster lung (CHL) cell line expressing human UGT1A9. METHODS: cDNA of UGT1 A9 was transcripted from mRNA by reverse transcriptase-...AIM: To clone the cDNA of UGT1A9 from a Chinese human liver and establish the Chinese hamster lung (CHL) cell line expressing human UGT1A9. METHODS: cDNA of UGT1 A9 was transcripted from mRNA by reverse transcriptase-ploymerase chain reaction, and was cloned into the pGEM-T vector which was amplified in the host bacteric E.Coli DH5(alpha). The inserted fragment, verified by DNA sequencing, was subcloned into the Hind III /Not I site of a mammalian expression vector pREP9 to construct the plasmid termed pREP9-UGT1A9. CHL cells were transfected with the resultant recombinants, pREP9-UGT1A9, and selected by G418 (400 mg x L(-1)) for one month. The surviving clone (CHL-UGT1A9) was harvested as a pool and sub-cultured in medium containing G418 to obtain samples forUGT1A9 assays. The enzyme activity of CHL-UGT1A9 towards propranolol in S9 protein of the cell was determined by HPLC. RESULTS: The sequence of the cDNA segment cloned, which was 1666 bp in length, was identical to that released by Gene Bank (GenBank accession number: AF056188) in coding region. The recombinant constructed, pREP9-UGT1A9, contains the entire coding region, along with 18 bp of the 5' and 55 bp of the 3' untranslated region of theUGT1A9 cDNA, respectively. The cell lines established expressed the protein of UGT1A9, and the enzyme activity towards propranolol in S9 protein was found to be 101+/- 24 pmol x min(-1) x mg(-1) protein (n=3), but was not detectable in parental CHL cells. CONCLUSION: The cDNA of UGT1A9 was successfully cloned from a Chinese human liver and transfected into CHL cells. The CHL-UGT1 A9 cell lines established efficiently expressed the protein ofUGT1A9 for the further enzyme study of drug glucuronidation.展开更多
phytohemagglutinin(PHA)-induced swelling is widely used to investigate cell-mediated and innate immunity across different vertebrate taxa.However,its physiological mechanism is still an open question due to the comple...phytohemagglutinin(PHA)-induced swelling is widely used to investigate cell-mediated and innate immunity across different vertebrate taxa.However,its physiological mechanism is still an open question due to the complexity of the involved immune components.In the present study,we measured the synchronous variations of PHA response,the proportion of different subtypes of leukocytes,as well as serum bactericidal capacity in circulation blood at 6,12 and 24 h after PHA versus PBS injection in striped hamster,Cricetulus barabensis.First,the results showed that PHA responses reached a peak at 6 h postinjection,then sharply declined at 12 h and 24 h postinjection.Serum bactericidal capacity was higher at 6 h and 12 h than at 24 h.The proportion of different subtypes of leukocytes,as well as the ratio of neutrophils to lymphocytes did not display significant changes across different time points.Second,PHA response was positively correlated with the proportion of neutrophils and serum bactericidal capacity.The proportion of monocytes was negatively correlated with that of eosinophils and neutrophils.The proportion of basophils was negatively correlated with that of lymphocytes.Our results indicate that earlier enhanced PHA response is important for the striped hamster to cope with changing environmental conditions due to its small body mass,and the increased components of innate immunity in circulation blood may contribute to the enhancement of PHA swelling response.展开更多
基金This work was supported by the National NaturalScience Foundation of China (Grant No. 39730090) the Key Projects of the Chinese Academy of Sciences (Grant Nos. KSCX2-1-03 and KZ951-B1-106).
文摘The impact of habitat fragmentation and isola-tion on the genetic diversity of populations has attracted much attention in studies of meta-population and conserva-tion biology. In this work, using the randomly amplified po-lymorphic DNA (RAPD) technique, we studied the genetic diversity of central, peripheral and peninsular populations of ratlike hamster, which were collected in five locations of the North China Plain and its surrounding areas, in 1999. The study revealed that, i ) the genetic diversity of central population of Raoyang County 】 the sub-central populations of Gu’an County and Taikang County 】 the peripheral population of Shunyi District 】 the peninsular population of Mentougou District; ii) the genetic diversities of the five populations were positively correlated to the nearest dis-tances to the peripheral line of population distribution; iii) there were significant differences of gene frequencies of some RAPD fragments among the five populations. More RAPD fragments disappeared in
基金Supported by the National Natural Science Foundation of China(C39370805)Zhejiang Provincial Natural Science Foundation(300487)the Excellent Youth Scientist Fund of Zhejiang Province
文摘AIM: To clone the cDNA of UGT1A9 from a Chinese human liver and establish the Chinese hamster lung (CHL) cell line expressing human UGT1A9. METHODS: cDNA of UGT1 A9 was transcripted from mRNA by reverse transcriptase-ploymerase chain reaction, and was cloned into the pGEM-T vector which was amplified in the host bacteric E.Coli DH5(alpha). The inserted fragment, verified by DNA sequencing, was subcloned into the Hind III /Not I site of a mammalian expression vector pREP9 to construct the plasmid termed pREP9-UGT1A9. CHL cells were transfected with the resultant recombinants, pREP9-UGT1A9, and selected by G418 (400 mg x L(-1)) for one month. The surviving clone (CHL-UGT1A9) was harvested as a pool and sub-cultured in medium containing G418 to obtain samples forUGT1A9 assays. The enzyme activity of CHL-UGT1A9 towards propranolol in S9 protein of the cell was determined by HPLC. RESULTS: The sequence of the cDNA segment cloned, which was 1666 bp in length, was identical to that released by Gene Bank (GenBank accession number: AF056188) in coding region. The recombinant constructed, pREP9-UGT1A9, contains the entire coding region, along with 18 bp of the 5' and 55 bp of the 3' untranslated region of theUGT1A9 cDNA, respectively. The cell lines established expressed the protein of UGT1A9, and the enzyme activity towards propranolol in S9 protein was found to be 101+/- 24 pmol x min(-1) x mg(-1) protein (n=3), but was not detectable in parental CHL cells. CONCLUSION: The cDNA of UGT1A9 was successfully cloned from a Chinese human liver and transfected into CHL cells. The CHL-UGT1 A9 cell lines established efficiently expressed the protein ofUGT1A9 for the further enzyme study of drug glucuronidation.
基金This study was supported by a Key Project of Natural Science of the Anhui Province Department of Education,China(KJ2011A114)supported by the State Key Laboratory of Integrated Management of Pest Insects and Rodents,Institute of Zoology,The Chinese Academy of Sciences(ChineseIPM1001)to ZZQ.
文摘phytohemagglutinin(PHA)-induced swelling is widely used to investigate cell-mediated and innate immunity across different vertebrate taxa.However,its physiological mechanism is still an open question due to the complexity of the involved immune components.In the present study,we measured the synchronous variations of PHA response,the proportion of different subtypes of leukocytes,as well as serum bactericidal capacity in circulation blood at 6,12 and 24 h after PHA versus PBS injection in striped hamster,Cricetulus barabensis.First,the results showed that PHA responses reached a peak at 6 h postinjection,then sharply declined at 12 h and 24 h postinjection.Serum bactericidal capacity was higher at 6 h and 12 h than at 24 h.The proportion of different subtypes of leukocytes,as well as the ratio of neutrophils to lymphocytes did not display significant changes across different time points.Second,PHA response was positively correlated with the proportion of neutrophils and serum bactericidal capacity.The proportion of monocytes was negatively correlated with that of eosinophils and neutrophils.The proportion of basophils was negatively correlated with that of lymphocytes.Our results indicate that earlier enhanced PHA response is important for the striped hamster to cope with changing environmental conditions due to its small body mass,and the increased components of innate immunity in circulation blood may contribute to the enhancement of PHA swelling response.
