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1.
Chinese Journal of Biotechnology ; (12): 375-383, 2015.
Article in Chinese | WPRIM | ID: wpr-240636

ABSTRACT

Taxa-4(5),11(12)-diene is the precursor for paclitaxel biosynthesis. The diterpenoid paclitaxel (marketed as Taxol), a plant secondary metabolite isolated from yew, is an effective drug widely used in the treatment of numerous cancers. However, further application of taxol has been restricted due to its low yield in plants and the difficulties in extraction. To increase the intact isoprene flux, we constructed the fusion gene plasmid pBgGGTS and individual cassette plasmid pBgGGgTS to enhance the expression levels of geranylgeranyl diphosphate synthase gene (ggpps) and a taxadiene synthase gene (ts) in Coprinopsis cinerea. These two plasmids were separately transformed into C. cinerea LT2 strain, resulting in several putative transformants. Putative transformants were determined by PCR technique, indicating that 5 out of 13 putative transformants transformed by pBgGGTS and 6 out of 13 putative transformants transformed by pBgGGgTS, respectively. Additionally, the Southern blotting analysis of these 10 transformants confirmed that both ggpps and ts gene were stably integrated into the genome of C. cinerea. Crude extracts from each of the transformants were analyzed. There is no difference in the mycelium extracts among the wild-type LT2 and two types of transformants. However, analysis of culture filtrates indicated that an additional GC peak was found at the retention time of 16.762 min which was absent in the wild type control. The mass fragmentation pattern of this peak had the same diagnostic ions with taxa-4(5),11(12)-diene. According to peak area, the amounts of taxa-4(5),11(12)-diene in each fermented broth were 44 ng/L (transformed with pBgGGgTS) and 30 ng/L (transformed with pBgGGTS), respectively. In conclusion, co-expression of the ggpps and ts gene could increase the taxadiene production in C. cinerea.


Subject(s)
Agaricales , Metabolism , Alkenes , Metabolism , Diterpenes , Metabolism , Farnesyltranstransferase , Genetics , Metabolism , Genetic Engineering , Isomerases , Genetics , Metabolism , Paclitaxel , Plasmids
2.
Journal of Practical Stomatology ; (6): 57-62, 2015.
Article in Chinese | WPRIM | ID: wpr-462130

ABSTRACT

Objective:To study the association between non-syndrome cleft lip and palate (NSCL/P)and MAFB (rs6072081, rs6065259,rs17820943,rs11696257)gene polymorphism in NingXia Hui and Han population.Methods:512 cases of NSCL/P,258 case of Hui nationality and 254 of Han,174 cases of complete 3 core family members(patients and their parents),142 cases of single parents families(patients and their single parents),were collected in patient group.505 cases of healthy newborns were collected in control group.The MAFB gene SNPs were determined with TaqMan SNPs genotyping methods and the data were analyzed by case-con-trol analysis,transmission disequilibrium test(TDT)and family based association test(FBAT).Results:Case-control analysis found that,there was a statistical significance of the genotypes and allele frequency between patients and the controls in MAFB of the 4 locus (P 0.05).Conclusion:The 4 locus single nucle-otide polymorphism was associated with NSCL/P in Ningxia population.

3.
West China Journal of Stomatology ; (6): 372-376, 2013.
Article in Chinese | WPRIM | ID: wpr-336316

ABSTRACT

<p><b>OBJECTIVE</b>To provide basis for effects of an early psychological intervention on parents of children with cleft lip/palate, and investigate the effects of an early psychological intervention to them.</p><p><b>METHODS</b>One self-administered questionnaire (SCL-90) was applied in 102 parents of children with cleft lip/palate, compared to 126 parents of healthy individuals on the day of admission. They were given the psychological intervention during hospitalization and 3 months after discharge. The questionnaire (SCL-90) was again applied to them on the day of discharge and 3 months after discharge.</p><p><b>RESULTS</b>Using the questionnaire (SCL-90), the answer scores of somatization, obsessive-compulsive, depression and anxiety etc. were significantly higher than those of the control group (P < 0.05). There were no statistical differences during 3 subgroups in the case group (P > 0.05). There were no statistical differences on the day of admission and on the day of discharge (P > 0.05), but there were statistical difference on the day of admission and 3 months after discharge (P < 0.05).</p><p><b>CONCLUSION</b>In this study, the psychosomatic states of parents of children with cleft lip/palate is poor. It's important and greatly significant that we conduct early psychological intervention to parents of children with cleft lip/palate and to the children's psychosomatic health.</p>


Subject(s)
Child , Female , Humans , Male , Cleft Lip , Cleft Palate , Parents
4.
West China Journal of Stomatology ; (6): 397-402, 2013.
Article in Chinese | WPRIM | ID: wpr-336310

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the association between rs142167, rs7216231 single nucleotide polymorphism (SNP) in Wnt3 and nonsyndromic cleft lip and palate (NSCL/P) in Hui and Han population of Ningxia Autonomous Region.</p><p><b>METHODS</b>The study consisted of 371 NSCL/P patients from Ningxia Hui and Han population (Han population 166, Hui population 205), their parents (196 fathers, 224 mothers, 150 trios) and 258 normal controls (Han population 190, Hui population 68). Polymerase chain reaction-restriction fragment length polymorphisms (PCR-RFLP) was used to identify rs142167, rs7216231 genotypes of the samples. The data was analyzed by case-control analysis, transmission disequilibrium test (TDT) and family based associated test (FBAT).</p><p><b>RESULTS</b>Case-control study showed that no differences in cleft lip, cleft palate, cleft lip and palate, and the total case group compared with the control group at rs142167 and rs7216231 (P > 0.05) in Hui and Han population and in stratified comparison. TDT test showed that rs142167 and rs7216231's allele had not over-transmitted (P > 0.05) in NSCL/P. FBAT test showed that G-G specific haplotypes showed statistically significant (P < 0.05).</p><p><b>CONCLUSION</b>Wnt3 gene polymorphism is not relevant with NSCL/ P in Ningxia Hui and Han population.</p>


Subject(s)
Humans , Brain , Congenital Abnormalities , Case-Control Studies , Cleft Lip , Cleft Palate , Genotype , Polymorphism, Genetic , Polymorphism, Single Nucleotide
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