1.Research advances of diterpene synthase.
Yunfei HE ; Wei GAO ; Tasi LIU ; Wenyuan LI ; Luqi HUANG
Acta Pharmaceutica Sinica 2011;46(9):1019-25
Diterpenes, an important class of natural compounds, are widely distributed in nature. As the valuable diterpenoids continue to be found, diterpene synthase in the course of diterpene synthesis get as much attention as possible. The multiformity of end-product-diterpenoids were also due to the diversity of diterpene synthase. This paper focuses on the advances in recent biosynthesis pathway of diterpene and types, cloning, catalytic mechanism, synthetic biology application.
2.Investigation of Resources and Varieties of Coptis Crude Drug Original Plant on Chinese Market
Xin LIU ; He HUANG ; Luqi HUANG ; Hezhen WU ; Yanfang YANG
China Pharmacist 2014;(10):1691-1695
Objective::To understand the resources distribution and varieties of crude drug Coptis chinensis Franch. on Chinese market to provide scientific basis for the resource conservation and variety collection and studies of the Chinese medicine. Methods:On the basis of literature survey, field investigation, mandatory survey and interview survey were conducted. Results:Due to the chan-ges in environment for many years and the predatory development in Coptis resources at the end of 20th century, the ecological environ-ment of wild Coptis resources changed significantly with destructed resources and decreased reserves, especially wild Yalian and Yun-lian were on the verge of extinction. The mainstream product on Chinese market was Weilian followed by Yunlian mainly used in small-scale medical clinic and medicine market in the distribution area, while Yalian commodity medicine could not found. Conclusion::The supply and demand on Coptis market should be stabilized to ensure herbalist interests. The production technology of Coptis need further improvement, cultivated area and suitable variety should be confirmed to guarantee the quality of Coptis medicinal materials. State should pay attention to the changes in Coptis resources and strengthen protection for wild Coptis resources to ensure the sustainable development of Coptis resources. Relevant government departments should guide the production of Coptis reasonably.
3.Cloning and induced expression analysis of 4-hydroxy-3-methyl-but-2-enyl diphosphate reductase gene (smHDR) of Salvia miltiorrhiza.
Qiqing CHENG ; Yunfei HE ; Geng LI ; Chao JIANG ; Yuan YUAN ; Wei GAO ; Luqi HUANG
Acta Pharmaceutica Sinica 2013;48(2):236-42
This study reported the obtainment of the full-length cDNA of Salvia miltiorrhiza hairy roots (Abbr: SmHDR, GenBank number: JX233817), via extracting Salvia miltiorrhiza hairy roots total RNA, designing specific primers according to the transcriptome data and using the RACE strategy, and then analyzed it with bioinformatics approaches. On this basis, using the real-time PCR to detect SmHDR gene expression after Ag+ induction, and testing tanshinones contents of corresponding samples by UPLC. SmHDR has 1 647 nucleotides, and an open reading frame (ORF) encoding a protein of 463 amino acid residues. The deduced protein has isoelectric point (pI) of 5.72 and a calculated molecular weight about 51.88 kD. In the secondary structure, the percentage of alpha helix, beta turn and random coil were 35.64%, 20.30% and 44.06%, respectively. Sequence alignment and phylogenetic analysis demonstrated that SmHDR had relative close relationship to the HDR of Picrorhiza kurrooa, similar to HDR from other species of plants. Real time PCR results indicated that elicitor of Ag+ stimulated the increase of mRNA expression of SmHDR. At the same time, results of ultra performance liquid chromatography (UPLC), used to examine the accumulation of diterpenoid tanshinones in hairy roots, showed that the contents of diterpenoid tanshinones in hairy roots of Salvia miltiorrhiza were increased dramatically at 12 h after treated with Ag+, and then decreased significantly. This result showed a positive correlation between the levels of mRNA expression and tanshinones accumulation in Salvia miltiorrhiza stimulated by Ag+. The content of tanshinones was gradually raised, and it had an obvious increase at 120 h. The bioinformatics analysis and gene expression indicated that SmHDR might be involved in tanshinones biosynthesis, which laid the foundation for further study of secondary metabolic regulation mechanism of tanshinones.
4.Investigate of DNA extraction of os cervi.
Jingxue ZHAO ; Min CHEN ; Guanghong CUI ; Shihuan TANG ; Luqi HUANG ; Liqun HE ; Ruixue XIA
China Journal of Chinese Materia Medica 2011;36(3):370-374
OBJECTIVETo establish a convenient, practical and high efficient method of DNA extraction of os cervi, and lay the foundation of identification of animal bones.
METHODThe bones of sika deer, red deer, cattle, dog and pig were used to extract DNA under different decalcification time (24,48,72 h) and decalcification temperature (4,25,37,56,70 degrees C), and extract method.
RESULTIt proved by experiments that demineralization process promotes the cracking of osteocyte. In a broad of decalcification time and temperature, DNA could be extracted from all bone samples successfully while the quantity varied slightly.
CONCLUSIONSamples (about 0.1 g) decalcify with 0. mol x L(-1) EDTA at 4 degrees C for 24 h, then water-bath for 1 h after lysis buffer added, DNA extracted via the method above is of high quality and can be used for PCR.
Animals ; Bone Demineralization Technique ; Bone and Bones ; chemistry ; metabolism ; Cattle ; DNA ; isolation & purification ; Deer ; Dogs ; Polymerase Chain Reaction ; Swine ; Temperature ; Time Factors
5.Effect of elicitors on induction and manipulation of secondary metabolic effective ingredients in Salvia miltiorrhiza.
Wenyuan LI ; Wei GAO ; Aijuan SHAO ; Yunfei HE ; Luqi HUANG
China Journal of Chinese Materia Medica 2011;36(3):258-262
The application of elicitors, which is currently the focus of research, has been considered as one of the most effective methods to improve the synthesis of secondary metabolites in medicinal plants. Biotic and abiotic elicitors can regulate the secondary metabolic pathways of effective ingredients in Salvia miltiorrhiza. This paper has introduced the research progress about the induction and the regulation mechanism of S. miltiorrhiza by elicitors.
Biosynthetic Pathways
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drug effects
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Drugs, Chinese Herbal
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pharmacology
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Salvia miltiorrhiza
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drug effects
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metabolism
6.Study on feasibility of cutting process of fresh Angelica sinensis Radix.
Liying TANG ; Zhuju WANG ; Bingsheng SONG ; Ziqing HE ; Hongwei WU ; Luqi HUANG
China Journal of Chinese Materia Medica 2010;35(23):3147-3150
OBJECTIVETo study on the feasibility of cutting process of fresh Angelica sinensis.
METHODQualitative and quantitative chemical analysis methods were used to evaluate the quality of different cutting processed A. sinensis.
RESULTThe contents of ligustilide and ferulic acid in the fresh cutting processed were both lower than the traditional cutting process, and the similarity of fingerprints of two different cutting processed A. sinensis were basically above 90%.
CONCLUSIONThe method of cutting process of fresh A. sinensis was not suitable apparently, and the effect on clinical application of these two different cutting processed A. sinensis need more study.
4-Butyrolactone ; analogs & derivatives ; analysis ; Angelica sinensis ; chemistry ; Chromatography, High Pressure Liquid ; Coumaric Acids ; analysis ; Drugs, Chinese Herbal ; analysis ; Plant Roots ; chemistry ; Technology, Pharmaceutical ; methods
7.Mechanisms of Intestinal Microecology in Hyperuricemia and Traditional Chinese Medicine Intervention:A Review
Mingyuan FAN ; Jiuzhu YUAN ; Hongyan XIE ; Sai ZHANG ; Qiyuan YAO ; Luqi HE ; Qingqing FU ; Hong GAO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(5):329-338
In recent years, hyperuricemia (HUA) has shown a rapidly increasing incidence and tends to occur in increasingly young people, with a wide range of cardiac, renal, joint, and cancerous hazards and all-cause mortality associations. Western medicine treatment has limitations such as large liver and kidney damage, medication restriction, and easy recurrence. The intestine is the major extra-renal excretion pathway for uric acid (UA), and the intestinal microecology can be regulated to promote UA degradation. It offers great potential to develop UA-lowering strategies that target the intestinal microecology, which are promising to provide safer and more effective therapeutic approaches. Traditional Chinese medicine (TCM) can treat HUA via multiple targets and multiple pathways from a holistic view, with low toxicity and side effects. Studies have shown that intestinal microecology is a crucial target for TCM in the treatment of HUA. However, its specific mechanism of action has not been fully elucidated. Focusing on the key role of intestinal microecology in HUA, this review explores the relationship between intestinal microecology and HUA in terms of intestinal flora, intestinal metabolites, intestinal UA transporters, and intestinal barriers. Furthermore, we summarize the research progress in TCM treatment of HUA by targeting the intestinal microecology, with the aim of providing references for the development of TCM intervention strategies for HUA and the direction of future research.
8.Analysis on Quality of Sojae Semen Praeparatum Based on Traditional Quality Evaluation
Yihan WANG ; Wangmin LIN ; Shuili ZHANG ; Bing YU ; Tiegui NAN ; Liping KANG ; Guofeng LI ; Xiyuan HE ; Zhilai ZHAN ; Luqi HUANG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(1):31-42
ObjectiveBased on the quality evaluation experience of "it is better to have a fragrant and strong aroma" summarized by materia medica of past dynasties, the chemical components of Sojae Semen Nigrum(SSN) and Sojae Semen Praeparatum(SSP) were systematically compared and analyzed, and the main fermentation products in different fermentation time were quantitatively analyzed, so as to clarify the transformation law of internal components in the processing process and provide scientific basis for the modern quality control of SSP. MethodUltra performance liquid chromatography-quadrupole tandem time-of-flight mass spectrometry(UPLC-Q-TOF-MS) was used for the structural identification of the chemical constituents of SSN and SSP, and with the aid of Progenesis QI v2.3 software, the negative ion mode was employed for principal component analysis(PCA) pattern recognition, and the data were analyzed with the aid of orthogonal partial least squares-discriminant analysis(OPLS-DA) for two-dimensional data to obtain S-plot, and components with |P|>0.1 were selected as the differential constituents. The contents of isoflavonoids in SSP during fermentation was determined by UPLC, and the samples were taken every 8 h in the pre-fermentation period and every 2 d in the post-fermentation period, and the dynamic changes of isoflavonoid contents in different fermentation stages were analyzed. The contents of amino acids and nucleosides in SSP and SSN from different fermentation stages were quantitatively analyzed by phenyl isothiocyanate(PITC) pre-column derivatization and high performance liquid chromatography(HPLC) gradient elution, and the contribution of flavor substances to the "delicious" taste of SSP was discussed by taste intensity value(TAV). ResultA total of 19 kinds of differential components were screened out, mainly soybean saponins and isoflavones, and their contents decreased significantly or even disappeared after fermentation. In the pre-fermentation process of SSP, glycoside bond hydrolysis mainly occurred, and isoflavone glycosides in SSN were degraded and converted into the corresponding aglycones, the content of flavor substances such as amino acids increased gradually. In the post-fermentation process, protein degradation mainly occurred, after 8 d of post-fermentation, the content of isoflavones was basically stable, while the total content of amino acids increased by 8-40 times on average. Different amino acids form the special flavor of SSP, such as the TAV of glutamate is always ahead of other flavor substances, and sweet substances such as alanine and valine have made relatively great contributions to SSP. ConclusionBased on the law of constituent transformation, combined with the traditional evaluation index of "fragrant and strong", it is difficult to control the fermentation degree of SSP by the existing standards in the 2020 edition of Chinese Pharmacopoeia. It is suggested that description of the characteristics of SSP be refined and changed to "fragrant, delicious and slightly sweet", and at the same time, the post-fermentation index compounds such as glutamic acid, alanine and valine should be added as the quality control indicators of SSP, so as to standardize the production process and improve the quality of SSP.