Polymorphism analysis of TCP gene region to intraspecific analysis of Paeonia lactiflora, and application for authentication of Paeoniae Radix

Author(s):  
Yuina Yoshie ◽  
Hirokazu Ando ◽  
Takayuki Tamura ◽  
Kozo Fukuda ◽  
Motoko Igarashi ◽  
...  
2021 ◽  
Vol 2021 ◽  
pp. 1-12
Author(s):  
Menglan Shen ◽  
Qiaoyan Zhang ◽  
Luping Qin ◽  
Binjun Yan

Paeoniae Radix Alba (PRA), an herbal drug produced from the root of Paeonia lactiflora Pall., is widely used in many herbal medicine prescriptions/preparations. Since the pharmacological effects of PRA come from multiple chemical components, it is important to establish a method for the determination of those components in PRA extracts with simple operation and low cost, which is more suitable to evaluate the quality of PRA extracts and optimize the extraction process. This work introduced the quantitative analysis of multicomponents with a single-marker (QAMS) method for the simultaneous determination of eleven bioactive components in PRA extracts, including gallic acid, oxypaeoniflorin, catechin, albiflorin, paeoniflorin, ethyl gallate, galloylpaeoniflorin, pentagalloylglucose, benzoic acid, benzoylpaeoniflorin, and paeonol. In the QAMS method established based on high performance liquid chromatography with diode array detection, only the reference substance of paeoniflorin was needed, and the other ten components were determined based on their relative correction factors (RCFs) to paeoniflorin. Moreover, the repeatability and robustness of the RCFs were studied with different column temperatures, detection wavelengths, flow rates, column types, and instruments. In method validation, good linearity (r > 0.999), stability, repeatability (RSD < 1.9%), and accuracy (recoveries within 96.1%–105.5%) were shown. Sample analyses showed that the QAMS method was consistent with the conventional external standard method. The established method provided a comprehensive, efficient, and low-cost tool for the routine quality evaluation of PRA extracts.


2009 ◽  
Vol 57 (9) ◽  
pp. 971-974 ◽  
Author(s):  
Kazuto Washida ◽  
Yoshiyuki Itoh ◽  
Takashi Iwashita ◽  
Kyosuke Nomoto

1996 ◽  
Vol 75 (06) ◽  
pp. 870-876 ◽  
Author(s):  
José Manuel Soria ◽  
Lutz-Peter Berg ◽  
Jordi Fontcuberta ◽  
Vijay V Kakkar ◽  
Xavier Estivill ◽  
...  

SummaryNonsense mutations, deletions and splice site mutations are a common cause of type I protein C deficiency. Either directly or indirectly by altering the reading frame, these' lesions generate or may generate premature stop codons and could therefore be expected to result in premature termination of translation. In this study, the possibility that such mutations could instead exert their pathological effects at an earlier stage in the expression pathway, through “allelic exclusion” at the RNA level, was investigated. Protein C (PROC) mRNA was analysed in seven Spanish type I protein C deficient patients heterozygous for two nonsense mutations, a 7bp deletion, a 2bp insertion and three splice site mutations. Ectopic RNA transcripts from patient and control lymphocytes were analysed by RT-PCR and direct sequencing of amplified PROC cDNA fragments. The nonsense mutations and the deletion were absent from the cDNAs indicating that only mRNA derived from the normal allele had been expressed. Similarly for the splice site mutations, only normal PROC cDNAs were obtained. In one case, exclusion of the mutated allele could be confirmed by polymorphism analysis. In contrast to these six mutations, the 2 bp insertion was not associated with loss of mRNA from the mutated allele. In this case, cDNA analysis revealed the absence of 19 bases from the PROC mRNA consistent with the generation and utilization of a cryptic splice site 3’ to the site of mutation, which would result in a frameshift and a premature stop codon. It is concluded that allelic exclusion is a common causative mechanism in those cases of type I protein C deficiency which result from mutations that introduce premature stop codons


2020 ◽  
Vol 86 (5) ◽  
pp. 480-485
Author(s):  
Lior Segev ◽  
Ilana Naboishchikov ◽  
Diana Kazanov ◽  
Ezra Bernstein ◽  
Meital Shaked ◽  
...  

Background CD24 is a sialoglycoprotein anchored to the cell surface via glycosylphosphatidylinositol and is involved in intracellular signaling processes. It plays an important role in the early stages of the multistep process of colorectal carcinogenesis. Several single nucleotide polymorphisms in the CD24 gene are reported to exert a diverse effect on cancer risk. We aimed to elucidate whether CD24 TG/del genetic variants are associated with susceptibility to colorectal cancer (CRC). Methods The study included 179 subjects, 36 with CRC (prior to surgery) and 143 healthy control subjects. Deoxyribonucleic acid was purified from peripheral blood leukocytes, and by using restriction fragment length polymorphism analysis, the CD24 gene was genotyped for the specific genetic variant, TG deletion. Additionally, CD24 protein expression levels were determined by Western blotting analysis. Results The incidence of the TG/del was higher among the CRC patients compared with healthy controls, 14% and 10%, respectively ( P = .54). CD24 protein levels were significantly higher among CRC patients. There were no significant differences in CD24 expression between CRC patients at different stages of the disease or between patients who carry the mutation and those who did not. Conclusions CD24 genetic variant might be of clinical value for risk assessment as part of cancer prevention programs. Further study on larger populations is needed to validate the importance of this dinucleotide deletion in CRC development. Overexpression of CD24 protein occurs early along the multistep process of CRC carcinogenesis, and a simple blood sample based on CD24 expression on peripheral blood leukocytes can contribute to early diagnosis.


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