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.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 .
3.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.
4.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.
5.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?.
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.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
8.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 .
9.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.
10.Effects of Ephedra on the body mass and levels of blood lipids, blood glucose and hormones in ovarietomized obese rats
Jinchao SI ; Xiaoyu ZHANG ; Meiling LIN ; Yonghui DING ; Wei LI ; Songyi QU ; Tianzhen ZHENG
Chinese Journal of Tissue Engineering Research 2006;10(47):209-212
BACKGROUND: Ephedra, a Chinese medicine, is often used to treat obesity with relatively satisfying results recently. However, the effects of Ephedra on the perimenopausal and postmenopausal obese women remain unclear.OBJECTIVE: To observe the effects of oral Ephedra decoction on body mass and the levels of blood lipids, blood glucose and hormone in ovarietomized obese rats.DESIGN: A completely randomized and controlled experiment.SETTING: Institute of Physiology and Psychology, School of Basic Medical Sciences, Lanzhou University.MATERIALS: The experiment was performed in the Key Laboratory of Pre-clinical Study for New Drugs of Gansu Province and the Laboratory of Institute of Physiology and Psychology, School of Basic Medical Sciences,Lanzhou University from February 2006 to June 2006. Forty-four healthy female SD rats were randomly divided into four groups with 11 rats in each group, namely sham-operated group, ovariectomized group, estrogen replacement therapy group and Ephedra group.METHODS: ① After having been narcotized by cloraminone (110 mg/kg),rats were underwent a bilateral ovariectomy except those in the sham-operated group, which were also operated, but their ovaries were not cut off. ②Rats in the sham-operated group and ovariectomized group were subcutaneously injected with sesame oil (0.2 mL/each rat) every day postoperatively till the end of the experiment. ③ The rats in the estrogen replacement therapy group were given estradiol (1 mg/kg) by subcutaneous injection every day postoperatively till the end of the experiment. ④ The rats in the Ephedra group freely drank 1% water extracts from Ephedra postoperatively, later the concentration of Ephedra gradually increased to 8% on the sixth day, which lasted until the end of the experiment. ⑤ The food intake was monitored daily, and body mass was measured every ten days. ⑥ At the end of the experiment, all the rats were fasted for 12 hours and collected blood samples for the measurement of serum indexes. The body mass and body length were measured to calculate the Lee's index [(g)×103/body length (cm)] at the same time.MAIN OUTCOME MEASURES: ① Body mass and Lee's index at different time points in each group. ② Food intake at different time points in each group. ③ Levels of blood lipids and blood glucose in each group. ④Levels of estrogen, progesterone and insulin in each group.RESULTS: Forty-four rats all entered the analysis of results. ① Result of body mass and Lee's index at different time points: The body masses on the 20th, 30th, 40th and 50th days in the ovariectomized group were (256.4±14.3),(271.3±16.1), (276.4±12.7), (285.7±24.2) g, which were significantly higher than those in the sham-operated group [(226.5±11.5), (241.8±12.6),(243.1±13.5), (251.1±22.4) g, P < 0.05-0.01], and the Lee's index in the ovariectomized group was greater than that in the sham-operated group(317.2±13.5, 280.4±11.2, P < 0.01). The body masses on the 40th and 50th days in the estrogen replacement therapy group were (243.7±14.8) and(246.2±11.9) g, which were significantly lower than those in the ovariectomized group (P < 0.05-0.01), and the Lee's index (289.9±13.5) was lower than that in the ovariectomized group (P < 0.01). The body masses on the 40th and 50th days in the Ephedra group were (245.4 ±14.1) and(252.4±14.9) g, and the Lee's index was 294.4±11.0, which were all lower than those in the ovariectomized group (P < 0.05). ② Result of Food in take at different time points: The food intakes on the 30th, 40th and 50th days in the Ephedra group were (17.8±2.4), (22.3±3.9), (26.1±3.5) g per day,which were decreased as compared with those in the ovariectomized group[(25.9±4.7), (28.5±5.3), (32.8±5.5) g per day, P < 0.05]. ③ Levels of blood lipids and blood glucose: The levels of triglyceride, cholesterol and low density lipoprotein cholesterol (LDL-C) in the ovariectomized group were (1.73±0.32), (1.45±0.50), (0.78±0.19) mmol/L, which were higher than those in the sham-operated group [(0.94±0.29), (1.05±0.30), (0.08±0.11) mmol/L, P < 0.01]. After the estrogen replacement therapy, the levels of triglyceride, cholesterol, LDL-C and blood glucose were (1.10±0.34),(1.14±0.30), (0.17±0.05), (5.88±1.21) mmol/L, which were lower than those in the ovariectomized group (P < 0.05-0.01), but the level of high density lipoprotein cholesterol (HDL-C) was higher than that in the ovariectomized group [(1.11±0.31), (0.88±0.21) mmol/L, P < 0.05]. The levels of triglyceride, cholesterol, LDL-C and HDL-C in the Ephedra group were (0.97±0.16), (1.11±0.20), (0.59±0.07) and (0.45±0.061) mmol/L, which were lower than those in the ovariectomized group (P < 0.05-0.01). ④ The serum levels of estrogen, progesterone and insulin in each group: The serum levels of estrogen and progesterone in the ovariectomized group were lower than those in the sham-operated group [(17.09±9.00), (28.51 ±7.99) μg/L;(58.69±12.11), (62.73±10.93) μg/L, P < 0.01], the serum level of insulin was higher than that in the sham-operated group [(31.74±6.69),(23.75±6.66) mU/L, P < 0.01]. The serum levels of estrogen in the estro gen replacement therapy and Ephedra group were (36.03±8.83) and (30.18±8.61) ng/L, which were higher than those in the ovariectomized group(P < 0.05-0.01), the level of insulin were (21.34±4.57), (24.86±6.20) mU/L,which were lower than those in the ovariectomized group (P < 0.05-0.01).The serum level of progesterone in the Ephedra group [(17.68±6.19) μg/L]was lower than that in the ovariectomized group (P < 0.01).CONCLUSION: Ephedra can promote loss of body mass, reduce levels of the blood lipids and insulin, and increase the serum levels of hormones in ovariectomized obese rats.