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1.
Acta Pharmaceutica Sinica ; (12): 2577-2583, 2021.
Article in Chinese | WPRIM | ID: wpr-886940

ABSTRACT

The molecular identification of Fritillaria taibaiensis and its relatives was studied by real-time PCR with a TaqMan-MGB probe. DNA was extracted from F. taibaiensis and its relatives. According to the sequence of ITS1 region, the mutation sites of F. taipaiensis and its related species were identified by MEGA7.0 software. The specific primers (a pair) and a TaqMan-MGB probe were designed by Primer Premier 6.0 software. In the Roche LightCycler 96 system, the lowest limit of detection for F. taipaiensis DNA template was 0.002 39 ng·μL-1, and the optimal Tm value range was 60 and 61 ℃. Specificity identification showed that the method had good specificity for F. taipaiensis, as it could be distinguished from other 13 different Fritillaria species including F. unibracteata. Since this method could accurately identify F. taipaiensis and its related species, it provides technical support for rational development of F. taipaiensis resources, management of Chinese medicinal market and supervision of raw materials in Chinese medicine manufacturing enterprises.

2.
Article in Chinese | WPRIM | ID: wpr-879004

ABSTRACT

Rhei Radix et Rhizoma is a kind of commonly used Chinese medicinal materials. Due to the overharvesting, the wild resource is endangering. Large market demand caused severely adulterant of commercial Rhei Radix et Rhizoma medicinal materials and decoction pieces. This manuscript reviewed the advances of the original species authentication in the industrial chain of Rhei Radix et Rhizoma during the latest decade, including characteristics and microscopic features, phytochemical analysis on anthraquinones, and molecular authentication based on DNA barcoding. Accordingly, an original species authentication route for the industrial chain of Rhei Radix et Rhizoma was summarized:(1)the identification of seeds and seedlings by DNA barcoding;(2) the selection of high variable sites based on the chloroplast genome;(3)biomonitoring of the Rhei Radix et Rhizoma medicinal materials and decoction pieces by two-dimensional DNA barcode;(4)traceability of Chinese patent medicines by third-generation sequencing. In conclusion, the combination of molecular identification and traditional identification methods provides a new idea for the identification of the original species of Rhei Radix et Rhizoma in the industrial chain and a essential guidance for the research of drug safety and efficacy of Rhei Radix et Rhizoma.


Subject(s)
Animals , Anthraquinones , Drugs, Chinese Herbal , Plant Roots , Rheum , Rhizome
3.
Acta Pharmaceutica Sinica ; (12): 2326-2334, 2019.
Article in Chinese | WPRIM | ID: wpr-780337

ABSTRACT

Based on the ITS2 and psbA-trnHsequences, molecular biological identification and genetic relationship of Fritillaria cirrhosa with its relative species were carried out. In this paper, the PCR-RFLP method specified by the Chinese Pharmacopoeia was performed on all samples at first. Secondly, the ITS2 and psbA-trnH sequences of all samples were amplified. Then, the amplified products were used to analyze the genetic distance, construct the phylogenetic tree, assess the identification efficiency, and evaluate the genetic relationship as well. The result showed that all the samples were divided into two groups by PCR-RFLP method. The samples in the first group, including Fritillaria ussuriensis, Fritillaria thunbergii and Fritillaria pallidiflora, could not be digested by SmaI, while the other samples in the second group, including Fritillaria mellea, Fritillaria sinica, Fritillaria cirrhosa var. ecirrhosa Franch, Fritillaria unibracteata var. longinectarea and Fritillaria cirrhosa, could be digested by SmaI. Then, ITS2 and psbA-trnH sequences of all samples were obtained. The length of various ITS2 sequences were distributed from 235 to 239 bp, and the average intra- and inter-specific genetic distance were 0.001 and 0.022, respectively. NJ tree showed that all samples were separated into "Northern Fritillaria" group (Fritillaria ussuriensis and Fritillaria pallidiflora) and "Southern Fritillaria" group (Fritillaria thunbergii, Fritillaria mellea, Fritillaria sinica, Fritillaria cirrhosa var. ecirrhosa Franch, Fritillaria unibracteata var. longinectarea and Fritillaria cirrhosa). The latter group could be further divided into Fritillaria thunbergii and Fritillaria cirrhosa subgroup, and the species in Fritillaria cirrhosa subgroup had close phylogenetic relationships. The length of psbA-trnH sequences was distributed from 337 to 373 bp, and the intra- and inter-specific genetic distance were 0.263 and 0.329, respectively. The samples in this paper could not be clustered effectively by NJ tree. This indicated that the ITS2 sequences were not only able to identify Fritillaria cirrhosa with its partial relative species quickly and accurately, but also clarify the relationship between different Fritillaria species. Therefore, it provided an important theoretical foundation for the development of molecular markers, effective protection, and rational development and utilization of Fritillaria resources.

4.
The Journal of Practical Medicine ; (24): 1995-1999,2003, 2018.
Article in Chinese | WPRIM | ID: wpr-697873

ABSTRACT

Objective To conduct a prospective phase Ⅱ clinical study to explore the distribution of CYP2D6 gene polymorphism in Chinese population and its relationship with the metabolism of tamoxifen in early-stage hormonal receptor-positive breast cancer. Methods CYP2D6 genotype was tested by Sanger sequencing using the ABI 3500 Genetic Analyzer. Plasma concentrations of tamoxifen and endoxifen were measured using the HPLC-MS/MS(API 2000)assay. We downloaded the data of CYP2D6 allele from the CPIP database. Results In Chi-nese patients,the most common alleles were CYP2D6*1,*2,and *10;the predominant diplotypes were *1/*10 (38.3%)and*10/*10(18.8%). The distribution of metabolic phenotype,plasma concentration of endoxifen,and endoxifen:tamoxifen plasma concentration ratio were inconsistent between the normal metabolic phenotype(EM) and the intermediate phenotype(IM)under different CYP2D6 activity prediction criteria.The differences in the ratios and endoxifen plasma concentrations were statistically significant between the three groups by cluster analysis. Conclusions The CYP2D6 genotype distribution in Chinese population is different from that in the Western popu-lation. There is considerable variation of serum endoxifen concentration in Chinese breast cancer patients possess-ing the phenotype previously known as the intermediate active metabolizers of CYP2D6. Therefore,in the current era of precision medicine,the standard CYP2D6 genotype-phenotype classification system cannot properly stratify the Chinese population with different levels of endoxifen plasma concentration.

5.
Article in Chinese | WPRIM | ID: wpr-616677

ABSTRACT

Objective To evaluate the value of DTI in mild articular cartilage injury in patellofemoral joint.Methods The DTI and arthroscopy data of 82 patients wih routine MRI diagnosed as mild articular cartilage injury were analyzed retrospectively.According to the results of arthroscopy,40 cases of mild articular cartilage injury with Outerbridge classification Ⅰ or Ⅱ were divided into experimental group,and 33 cases with normal patellofemoral articular cartilage were divided into control group.There were 8 articular cartilage injury patients with Outerbridge classification Ⅲ or Ⅳ in patello-femoral join were excluded.The DTI data were analyzed compared with arthroscopy.Results Arthroscopy detected 62 lesions of cartilage injury in experimental group.Totally 49 lesions (49/62,79.03 %) were detected by ADC pseudocolor image and 51 lesions (51/62,82.25 %) were detected by FA pseudocolor image.The DTI pseudocolor images of articular cartilage injury showed uneven levels.The red or pink levels can been observed.Compared with the control group,ADC value increased and FA value decreased significantly in experimental group (both P<0.05).Conclusion DTI can clearly display and detect mild articular cartilage injury in patellofemoral joint,which provide valuable information for early cartilaginous injury.

6.
Tianjin Medical Journal ; (12): 1208-1212, 2017.
Article in Chinese | WPRIM | ID: wpr-667912

ABSTRACT

Drug rash with eosinophilia and systemic symptoms (DRESS) syndrome is a serious adverse drug reaction, characterized with rash, fever, lymphadenectasis, eosinophilia and visceral involvement. This article describes the clinical case of a patient with renal insufficiency after receiving sensitizing drugs,which resulted in limb weakness and cognitive impairment of center nervous system characterized by vasculitis imaging and responded well to glucocorticoid treatment-DRESS syndrome.

7.
Article in Chinese | WPRIM | ID: wpr-577825

ABSTRACT

Objective To clone and sequence the cDNA encoding(S)-N-methylcoclaurine-3'-hydroxylase from Coptis chinensis.Methods The cDNA,encoding(S)-N-methylcoclaurine-3'-hydroxylase,was amplified by RT-PCR with cDNA library of tender leaf as the template.Results The full-length cDNA of(S)-N-methylcoclaurine-3'-hydroxylase(named as CYP80B3) had 1 680 bp with an open reading frame encoding 488 amino acids of protein.The CYP80B3 had 95%,82%,70%,and 68% amino acid sequence homology to the sequence of(S)-N-methylcoclaurine-3'-hydroxylase from C.japonica,Thalictrum flavum,Eschscholzia californica and Papaver somniferum,respectively.The sequence was reported to the GenBank and coded as EF492879.Comparison of sequence with(S)-N-methylcoclaurine-3'-hydroxylase from C.japonica showed CYP80B3 possessed the same functional regions involved with 3'-hydroxylation of(S)-N-methylcoclaurine.Conclusion The cDNA encoding CYP80B3 from C.chinensis was cloned and reported.This work underlays the first step for exploring the pathway of benzylisoquinoline alkaloid biosynthesis and for improving the content of benzylisoquinoline alkaloid in C.chinensis.

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