1.Different active ingredients of medicinal plant based on function differentiation of homologous gene.
Yuan YUAN ; Jun YU ; Lu-qi HUANG ; Xu-min WANG
China Journal of Chinese Materia Medica 2015;40(6):1023-1026
In the research field of quality control in Chinese medicinal materials, variation in active ingredients of medicinal plant is always the key and hot issues. With the development of high-throughput sequencing technologies and reducing cost, a large numbers of genes from medicinal plant were cloning and provide a solid foundation for further research of gene structure and its biological function, and also provides conditions for explore active ingredient variation and its quality control from the perspective of molecular pharmacognosy. This paper introduces the concept of homologous gene, gene duplication and classification. We prospect the function of duplicated genes in the role of molecular mechanism research about variation in active ingredients, aiming at providing a new way for medicinal materials quality control.
Drugs, Chinese Herbal
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analysis
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Gene Duplication
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Plant Proteins
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genetics
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Plants, Medicinal
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chemistry
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genetics
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Quality Control
2.Comparison of Several Methods and Their Progresses on Molecular Authentication of Lonicera japonica Thunb
Chao JIANG ; Luqi HUANG ; Yuan YUAN ; Shulin YU ; Min CHEN
World Science and Technology-Modernization of Traditional Chinese Medicine 2014;(8):1831-1839
Authentications of Chinese herbal medicine have a critical effect in Chinese clinical medicine. DNA molecular marker, as an important component for true or false authentication, is more and more widely used in iden-tification of Chinese medicinal materials. At the same time, many new methods for authentication of Chinese medici-nal materials are continuously emerging. But the systematically comparative analysis of these new methods is lack. The present study taking Lonicera japonica as an example, systematically compared principles, characteristics, ex-periment methods, detection time and the application scope of express sequence tag-simple sequence repeat (EST-SSR), polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), allele-specific PCR (AS-PCR), DNA barcoding and loop-mediated isothermal amplification (LAMP), and put forward corresponding improve-ment opinions. This study can help to screen appropriate approach for rapid authentication of L. japonica and offer demonstrating to other Chinese herbal medicines.
3.Stable expression of Hantavirus H8205 strain G1/IL-2 gene and immune protection of the fusion gene.
Ying, XIONG ; Yuan, YUAN ; Min, JIA ; Bing, YU ; Hanju, HUANG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2007;27(2):124-7
To explore the feasibility of stable expression of Hantavirus H8205 strain G1 segment and human IL-2 fusion gene in Vero cells, and to examine the immune protection effects on mice vaccinated with this recombinant eukaryotic expression vector containing Hantavirus G1 gene and IL-2 gene. With the help of lipofectamine, the Vero cells were transfected with pcDNA3.1/HisB-IL-2-G1 and the positive cells were selected by G418. IFAT and SDS-PAGE electrophoresis were used to determine the stable transfection and expression of recombinant protein. Each mouse was inoculated with plasmids intramuscularly (i.m.) three times, 2 boosts were given at 2-week intervals, serum anti-hantavirus antibodies were detected by ELISA and neutralizing antibodies (NAb) were detected by Plaque Reduction Neutralization Test. The fusion protein expressed in Vero cells was 78 kD, corresponding to the estimated molecular size. The neutralizing antibody titers of mice with pcDNA3.1/HisB-IL-2-G1 were 1:20-1:80. IL-2/G1 fusion gene could be transferred in Vero cells and stably express the fusion protein. Specific humeral immune responses in mice can be induced with the recombinant eukaryotic expression vector containing the fusion gene, which lays the foundation for further development of therapeutic HTNV vaccine.
4.Strategies of the phenomenon of college teachers' multimedia dependency
Juan WANG ; Fei JIAO ; Yuan YU ; Xinhua NIU ; Min YU
Chinese Journal of Medical Education Research 2011;10(7):819-821
Currently, multimedia teaching plays a more and more important role in colleges.However, the negative effects of multimedia teaching have gradually revealed because of the neglection of its limitations. In this article, strategies were proposed from three different aspects against disadvantages of multimedia teaching in colleges in order to improve its role more effectively.
5.Observation of extracellular matrix damage in the kidney of rats with multiple organ dysfunction syndrome
Yurong YUAN ; Lin TENG ; Min YU ; Xianzhe LIU
Chinese Journal of Emergency Medicine 2012;21(8):819-824
ObjectiveTo determine the levels of type Ⅳ collagen and matrix metalloproteinase-9 ( MMP-9 ) in the serum and kidney of rats with the multiple organ dysfunction syndrome ( MODS ) and investigate the mechanism of extracellular matrix damage in renal failure of MODS.MethodsForty adult male Sprague- Dawley (SD) rats were randomly (ramdom number) divided into two groups,namely the normal control group ( n =8 ) and the MODS model group ( n =32 ).The rats of model group were further divided into four sub-groups as per different intervals,6 h,12 h,24 h and 48 h,after modeling (n =8 in each).The animal models of MODS were established by two hits,the left eyeball of each model rat was removed to bleed to 2 ml bloocd/100 g body weight and lipopolysaccharide ( LPS,5 mg/kg) was injected into intraperitoneal cavity of model rats four hours later. The same volume of saline instead was injected intraperitoneally into rats of control group.All rats were sacrificed at different intervals.Creatinine (Cr)and blood urea nitrogen (BUN) were determined by using a Hitachi Automatic Biochemical Analyzer.The histological changes in renal tissue were observed under light microscope and transmission electronic microscope.The levels of serum type Ⅳ collagen and MMP - 9 protein were measured by using enzyme linked immunosorbent assay (ELISA).Type Ⅳ collagen and MMP- 9 protein levels in renal tissue were detected by western blot.One - way ANOVA was used for comparison among multiple groups.Results Compared with the control group,Cr and BUN were significantly higher in MODS group ( P <0.05 ).There were no histopathological changes in kidney of rats in control group,and the renal injury was serious in rats with MODS.The remarkable edema of basement membrane and defect of collagen fibers in renal tissue were observed in MODS group.Compared with the control group,the levels of MMP-9 in serum increased 6-48 hours after modeling and peaking at interval of 12 hours after modeling ( P < 0.05 ).The protein levels of type Ⅳ collagen increased not significantly in 6 h group and 12 h group in comparison with control group (P > 0.05 ),while those in 24 h group and 48 h group significantly increased ( P < 0.01 ).The levels of MMP -9 in renal tissue of rats with MODS increased in 6 h group and 12 h group (P<0.05),peaked in 24 h group ( P < 0.05 ),and decreased in 48 hours.However,the level of Ⅳ collagen in serum of rats with MODS decreased significantly 6-48 hours after modeling (P < 0.05 ) Conclusions The injury of extracellular matrix is an important factor to the kidney damage in MODS and it may he used for early diagnose and as a treatment target for kidney injury in MODS.
6.Perioperative management and impact of preoperative renal dysfunction on short-term survival for patients undergoing valve replacement
Jian LIU ; Yizhou YE ; Min YU ; Sheng SHI ; Zhongxiang YUAN
Chinese Journal of Thoracic and Cardiovascular Surgery 2013;(4):200-203
Objective To review the experience of perioperative management and effect of valve replacement combined with renal dysfunction.Methods 536 cases of valve replacement.According to preoperative glomerular filtration rate(GFR),renal dysfunction was classified as normal in 322 cases,mild in 162,moderate in 40 and severe in 12.Logistic regression analysis was used to assess the effect of preoperative renal dysfunction on operative mortality and morbidity.Results Patients with a lower GFR were older and more likely to have hypertention.They also tended to have larger left ventricular end-diastolic diameter and worse left ventricular ejection factor.Operative mortality rose inversely with declining renal function,from 2% for those with normal renal function to 17% for patients with severe renal dysfunction.Renal dysfunction was significantly associated with a large blood transfusions,re-intubation,longer mechanical ventilation,intensive care unit stay and hospital stay.Conclusion Preoperative renal dysfunction is an important factor of postoperative mortality and morbidity for valve replacement.We must pay more attention to perioperative management.
7.The expression pattern of pleiotrophin in colorectal malignancy
Yuan GAO ; Jiren YU ; Shusen ZHENG ; Min ZHANG ;
Chinese Journal of General Surgery 1994;0(05):-
ObjectiveTo study the expression of pleiotrophin (PTN) in colorectal cancer and its impact on the malignancy.MethodsRT PCR and immunoperoxidase staining were applied on the resected samples.ResultsPTN mRNA was expressed in 18 cases of 24 colorectal cancer tissues and 18 cases of 19 adjacent normal tissues. PTN protein was also found expressed in stroma, but the expression rate was higher in colorectal cancer tissues (34%) than in adjacent normal tissues (9%, P
9.EST-SSR identification of Lonicera japonica Thunb.
Chao JIANG ; Yuan YUAN ; Guiming LIU ; Luqi HUANG ; Xumin WANG ; Jun YU ; Min CHEN
Acta Pharmaceutica Sinica 2012;47(6):803-10
Simple and effective methods are needed for the identification of Chinese medicinal material species and their variety. Lonicera japonica Thunb. is one of Chinese herbal medicines widely demanded. A total of 3 705 EST-SSRs of L. japonica and 2 818 EST-SSRs of L. japonica var. chinensis Thunb. were identified from EST database in our lab. In average, there was one EST-SSR per 4.05 kb in L. japonica ESTs and per 7.49 kb in L. japonica var. chinensis ESTs, separately. The identified SSRs in L. japonica were consisted of 51.98% dinucleotide and 34.61% trinucleotide repeats, while SSRs in L. japonica var. chinensis had 57.45% dinucleotide and 30.09% trinucleotide. The results reviewed that the classes AG/TC and GAG/TCT were predominant in the dinucleotide motifs and the trinucleotide motifs, respectively. Total 87 EST-SSRs were identified of significant difference between L. japonica and L. japonica var. chinensis. PCR products were obtained from 52 L. japonica samples in 13 out of 15 SSR markers tested. The polymorphism in L. japonica, L. japonica var. chinensis and other honeysuckles could be distinguished by three markers (jp.ssr4, jp.ssr64 and jp.ssr65) tested.