1.Development of microsatellites of Dendrobium officinale and its application in purity identification of germplasm
Minglu XIE ; Beiwei HOU ; Li HAN ; Yanhong MA ; Xiaoyu DING
Acta Pharmaceutica Sinica 2010;45(5):667-72
Simple sequence repeat (SSR) was used to investigate the genetic diversity and structure of Dendrobium officinale. A total of 15 primer pairs with stable and repeatable polymorphism were screened out from 60 SSR primer pairs developed by the method of microsatellite enrichment by magnetic beads. Forty-eight samples of Dendrobium officinale were analyzed in genetic polymorphism. These loci were polymorphic and displayed 3 to 9 alleles per locus with a mean number of 6.1. The observed and expected heterozygosities ranged from 0.60 to 0.85 and from 0.49 to 0.85 respectively. The polymorphic information content (PIC) of each SSR locus varied from 0.437 to 0.829 with an average of 0.702. Fifteen primer pairs were used in Dendrobium cross-species amplification and totally 13 primer pairs were proved to have the transferability in D. officinale related species. In addition, 500 tissue culture plantlets of D. officinale were tested for purity identification by means of PCR amplification with four SSR primer pairs. The results showed that SSR technique is a feasible, simple and inexpensive method for determining adulterants in germplasm identification.
2.Sequence analysis of LEAFY homologous gene from Dendrobium moniliforme and application for identification of medicinal Dendrobium.
Wenrui XING ; Beiwei HOU ; Jingjiao GUAN ; Jing LUO ; Xiaoyu DING
Acta Pharmaceutica Sinica 2013;48(4):597-603
The LEAFY (LFY) homologous gene of Dendrobium moniliforme (L.) Sw. was cloned by new primers which were designed based on the conservative region of known sequences of orchid LEAFY gene. Partial LFY homologous gene was cloned by common PCR, then we got the complete LFY homologous gene Den LFY by Tail-PCR. The complete sequence of DenLFY gene was 3 575 bp which contained three exons and two introns. Using BLAST method, comparison analysis among the exon of LFY homologous gene indicted that the DenLFY gene had high identity with orchids LFY homologous, including the related fragment of PhalLFY (84%) in Phalaenopsis hybrid cultivar, LFY homologous gene in Oncidium (90%) and in other orchid (over 80%). Using MP analysis, Dendrobium is found to be the sister to Oncidium and Phalaenopsis. Homologous analysis demonstrated that the C-terminal amino acids were highly conserved. When the exons and introns were separately considered, exons and the sequence of amino acid were good markers for the function research of DenLFY gene. The second intron can be used in authentication research of Dendrobium based on the length polymorphism between Dendrobium moniliforme and Dendrobium officinale.
3.Studies on chemical constituents from root of Isatis indigotica Ⅰ
Yunhai LIU ; Guowei QIN ; Shuiping DING ; Xiaoyu WU
Chinese Traditional and Herbal Drugs 1994;0(12):-
Object To extract and separate the chemical constituents from the root of Isatis indigotica Fort (Cruciferae) Methods The root of I. indigotica was percolated with 95% ethyl alcohol, partitioned in solvents of different polarities and finally isolated on silica gel and macroporous resin columns The purified compounds obtained were identified and structurally elucidated by their physicochemical properties and spectral analysis Results Two compounds were obtained and named as isaindigotidione (Ⅰ) and (E)-3-(3′, 5′-dimethoxy-4′-hydroxybenzylidene)-2-indolinone Conclusion The two compounds were new
4.Consideration on Establishment of Physiochemical Quality Standard for Health Food
Weiqing ZHANG ; Jin CAO ; Hong DING ; Xiaoyu LI
China Pharmacist 2016;19(7):1352-1354
Under the enforcement background of the newly issued Food Safety Law and combined with the work on registration and record of health food , the article put forward the problems and challenges of health food regulation according to the physiochemical quality standard for health food .The paper provided methods and suggestions for the establishment of physical and chemical indices for health food , and discussed how to improve the compositions of physical and chemical testing indicators for health food and improve the specificity of the standard , which can provide basis and guarantee for the post-marketing supervision of health food .
5.Diagnosis significance of medical thoracoscopy in tuberculous pleural effusion
Xiaoyu ZHOU ; Gengyun SUN ; Wei LI ; Qiang DING ; Yuqing CHEN
Chongqing Medicine 2015;(22):3045-3047
Objective To assess the accuracy and safety of medical thoracoscopy(MT)in the diagnosis of tuberculous pleural effusion.Methods We evaluated 52 patients who were suspected tuberculous pleural effusion.The diagnosis rate and complications of medical thoracoscopy was assessed.Results About 33 of 52 patients were tuberculous pleural effusion.Twenty-nine cases were diagnosed by medical thoracoscopy,and the diagnostic rate was 88%.Under the thoracoscope,clinical manifestations of these pa-tients with tuberculous pleuritic were miliary nodules in 23 cases (70%),fiber cord-like adhesions in 12 cases (36%),extensive wrapped with fiber deposition in 7 cases(21%),and white scar in 5 cases (1 5%).All complication was relived or caused,and 1 case of gas embolism was the most serious one.Conclusion Medical thoracoscopy was a method with high diagnostic value and safety in the diagnosis of tuberculous pleural effusion.
6.Application of Quality Metrics in Food Inspection Laboratory Management
Huan HE ; Jin CAO ; Hong DING ; Yong LU ; Xiaoyu LI
China Pharmacist 2017;20(8):1457-1460
Because of the particularity and the complexity of integrated management of food, government regulators must perform the whole process of quality control and trace to evaluate and judge the merits of food instead of only depending on product standards.Quality metrics is a new concept for the food industry, and can be regarded as the comprehensive evaluation and measurement for product quality and the standard for judging the quality of food products.Request for Quality Metrics Guidance for Industry was issued, which not only established a new model for the quality control of pharmaceutical industry, but also provided a new way of quality management for the inspection organization.By introducing the contents of Request for Quality Metrics Guidance for Industry issued by the US FDA, on the basis of summarization and thought, the quality management and evaluation of pharmaceutical industry was compared, and the feasibility and applicability of quality metrics management in inspection and testing field, especially in food testing field was evaluated.In this paper, the concept of quality metrics was extended to the level of food inspection laboratory management with hope to provide ideas for the future development of food quality management model.
7.Identification of Trapa L. plants along middle-low reaches of Changjiang River by analyzing their DNA sequences
Shulin BAO ; Xiaoyu DING ; Jun CHANG ; Jie SHEN ; Feng TANG
Chinese Traditional and Herbal Drugs 1994;0(08):-
Object To analyze the rDNA ITS sequences between wi ld plants and cultivars of Trapa L. and study the utility in p hylogenesis and identification of these two groups. Methods The ITS gene fragments were PCR amplified and sequenced. The rDNA ITS regions w ere analyzed by means of the software of Clustal and Mega 2.0. Result s The rDNA sequences of 234-236 bp ITS1, 220-221 bp ITS2 gene fragment , and 5.8 S rDNA for 164 bp evenly were obtained from ten populations of Trapa L. The intraspecific substitution varies from 0.22% to 2. 94%. The variable sites are 16 while informative sites are six. The phylogenet ic tree based on ITS data was set up by NJ method. Conclusion ITS sequence is a pretty good molecular marker which can identify wild plants of Trapa L. from their cultivars. Diversity of ITS in differen t populations is less at intraspecific level. It is infered that the plants of Trapa L. may be derived from the same population of one species .
8.The diagnostic significance of CT tumor angioimaging on the peripheral lung cancer
Yanqing HUA ; Guozhen ZHANG ; Qiyong DING ; Guoxing NI ; Xiaoyu LU
Chinese Journal of Radiology 2001;0(07):-
2 cm).Immunohistochemistrical stain slice of operative specimen showed a lots of vascular endothelial cells and small vessels,which correlated with the tumor angioimaging area on CT images.The sensitivity,specificity,and positive predictive valve of tumor angioimaging in the lung cancers were 47.2%,91.3%,and 95.5%,respectively.Conclusion The “tumor angioimaging” sign can be regarded as another specific feature in peripheral lung cancer with contrast CT.
9.RNA interference inhibits the secretion of IL-1? in mice spleen lymphocytes
Ningning ZHENG ; Yanqiu YU ; Xiaoyu SONG ; Xudong DING ; Haipeng ZHANG
Chinese Journal of Pathophysiology 1986;0(03):-
AIM: To decide the effect that selected siRNA degrades mRNA of IL-1? specifically and suppression of its expression after connected with target site with homology complementary sequence. METHODS: Synthesized DNA expression box aimed directly at target site through PCR reaction in vivo was purified, and transfected into lymphocytes stimulated by LPS. siRNA was transcribed by cellular endogenous RNA polymerase Ⅲ and then evoke the degradation of target mRNA. After 48 hours of transfection, the cell culture supernatant was collected and the concentration of IL-1? was assayed using ELISA. RESULTS: Compared with blank-control and negative-control, selected sequence decreased the expression of IL-1?. Rate of the suppression was about 15%. CONCLUSION: RNAi technology produces specific interference effect in mouse spleen lymphocytes in original culture and inhibits the excretion of IL-1?.
10.Evaluation of genetic diversity of Dendrobium officinale wild populations based on RAMP markers.
Jie SHEN ; Huijun XU ; Yinghui YUAN ; Li HAN ; Beiwei HOU ; Xiaoyu DING
Acta Pharmaceutica Sinica 2011;46(9):1156-60
Random amplified microsatellite polymorphism (RAMP) markers were used to access the genetic diversity among 112 samples of nine populations of Dendrobium officinale Kimura et Migo. Using 16 informative primers, 123 bands were amplified and 86 (69.92%) were polymorphic. The polymorphic bands from three to eight could be detected for each RAMP primer, with a mean of 5, indicating abundant genetic diversity among populations. Genetic similarity coefficients ranged from 0.250 to 0.813. UPGMA dendrogram illustrated 9 populations clustered into 3 groups, and the cluster pattern showed correlation with the locations of the D. officinale populations. These results were supported by the previous conclusions that were achieved by other molecular markers, and RAMP is proved to be effective for evaluating the genetic diversity of wild populations of Dendrobium officinale.