A change in vascular smooth muscle cell (VSMC) phenotype, known as converting from a contractile state into a synthetic phenotype, is a crucial event in vascular remodeling, which determines the occurrence, development and prognosis of cardiovascular diseases such as pulmonary hypertension and atherosclerosis. Research shows that the expression level of key signaling molecules, which controls the phenotype change of VSMC, is regulated by microRNA (miRNA), a type of non-encoding RNA. In this article, we provide a review of miRNA in the regulation of VSMC phenotype changes with a focus on the key molecules in MAPK, TGFβ/Smad and PI3K/Akt signaling pathways.

Sab (SH3 domain-binding protein that preferentially associates with Btk) that is also called SH3BP5 (SH3 domain-binding protein 5), is a scaffold protein on mitochondrial outer membrane in the modulation of mitochondrial function. Sab not only combines with the tyrosine kinase Btk (Bruton's tyrosine kinase), but also binds to the serine threonine kinase JNKs (c-Jun amino-terminal kinases) and p38γ. Thus Sab can regulate B cell antigen receptor, mitochondrial JNK and p38γ signaling pathway, which is associated with the critical physiological function, such as B-cell development and differentiation and regulation of mitochondrial signaling transcription. Inhibition or induction on the expression of Sab can ameliorate the diseases arising from the abnormal level of Btk, JNKs and p38γ, such as nervous system diseases and liver injury. Therefore, Sab could be expected as a new target for drug development. In this article, we provide an overview of the structure and functions of Sab and its relationship to diseases of diffuse large B-cell lymphoma, nervous system diseases and liver injury, aiming to provide new ideas and theoretical basis for the development of new drugs.

Histone H3 lysine 79 (H3K79) methyltransferase DOT1L plays an important role in the activation and maintenance of gene transcription; it is also essential for maintenance of embryonic development, as well as the normal function of hematopoietic system, heart and kidney. However, the over expression of DOT1L is associated with the occurring and progress of numerous malignant tumors, so more and more attention has been paid to DOT1L. Therefore, it is of great significance to study and develop inhibitors of DOT1L. The inhibitors could serve as a tool in the investigation of the biological function, and have the potential to be developed into novel anti-cancer agents in the anticancer therapy. This paper mainly describes the structure and function of DOT1L, the relationship between DOT1L and tumors as well as the latest research progress of DOT1L inhibitors; with expect to provide some useful references for the subsequent research.

Apoptosis is an important self-stabilizing mechanism of multicellular organisms, which plays a vital role in the development of normal living organisms and the maintenance of tissue homeostasis. The abnormalities in apoptosis often lead to body lesions including tumors. Studies have shown that Bcl-2 protein with anti-apoptosis activity is an important target in the treatment of cancer. After nearly two decades of efforts, many small molecule Bcl-2 inhibitors have been discovered to induce cell apoptosis. The small-molecule Bcl-2 inhibitor, venetoclax, was developed based on fragment-based drug design strategy and approved by the FDA in 2016 for clinical application. This agent is the first approved small-molecule drug inhibiting Bcl-2 through protein-protein interaction to induce cell apoptosis. The achievement of venetoclax benefits from a combination of drug discovery technologies and represents a milestone in the history of drug discovery. In this review we will introduce the current progress in Bcl-2 inhibitors.

Gene therapy has obvious advantages in the treatment of ocular diseases due to the unique structure of the eye. In recent years, there are more and more therapeutic gene-based drugs for ophthalmic application in clinical trials. Most of the delivery vectors are adeno-associated virus and administered via intraocular injection, which has potential risks. Traditional remedies, such as topical instillationor systemic administration, have limited therapeutic effects on the diseases in the posterior segment of the eye, where the chemical drugs are hard to reach. This makes the research of new strategies for gene drug delivery extremely urgent. For better understanding of the latest hot topics of ocular gene therapy, this article is prepared to introduce application of gene therapy to the typical ocular diseases and the corresponding gene-based medicines. The absorption routes for gene delivery into eyes and existing barriers are summarized. Finally, the gene delivery strategies are highlighted. The clinical application of ocular gene therapy will be boosted by overcoming the absorbing barriers and reducing the potential pitfalls.

Nanocrystals is a hot topic of poorly soluble drug delivery system under development. Nanocrystals is different from the nanoparticles for drug unloading in polymer materials, and nanocrystals is different from solid dispersion system, in which the drug maintains crystals state. Nanocrystals has a simple preparation process, represents an effective technology in the improvement of solubility and bioavailability of poorly soluble drugs, and has a very promising industrialization application and development potential. In this paper, we retrospect the development history of drug nanocrystals technology, review development of the preparation methods of nanocrystals, and analyze the existing problems to provide a reference to the development of drug nanocrystals preparation.

Anthocyanin is a water-soluble flavonoid pigment which is widely found in plants. Studies showed that anthocyanin had protective effect on vision. However, whether anthocyanin has therapeutic effect on cataract remain unclear. In this study, we established the age-related and posterior capsule opacification cataract cell models through inducing oxidative damage of human lens epithelial cells (HLECs) by H₂O₂ and inducing epithelial-mesenchymal transition (EMT) by transforming growth factor β₂ (TGF-β₂). The preventative effects of anthocyanin on markers of oxidative damage and EMT were determined by respective assay kits and PCR analysis. Anthocyanin was beneficial to reduce oxidative stress of HLECs, protecting cells from H₂O₂ induced damage and increasing α-crystallin expression. The potential mechanisms might be that anthocyanin increased the activities of SOD and GSH-Px, which contributes to reduce cellular ROS and MDA level. Besides, anthocyanin inhibited Ca²⁺ overload, which contributes to protection of cell from apoptosis. Meanwhile, anthocyanin had inhibitory effect on EMT, slowed down cell proliferation, migration caused by TGF-β₂ through decreasing mRNA expression levels of EMT markers including COL1A1, COL1A2, COL3, COL4, Fn and α-SMA. The results suggest that anthocyanin could protect HLECs from oxidative damage induced by H₂O₂ and cell proliferation, migration and EMT induced by TGF-β₂, which indicated that anthocyanin may have protective and therapeutic effects on age-related cataract and posterior capsule opacification.

The impact of Rg1 in the disease progress and pathology of amyotrophic lateral sclerosis (ALS) was investigated in mouse model (SOD1 G93A). Body weight and survival rate were monitored to check the course of disease. Rotarod test was used to evaluate the coordination of muscle movement. Toluidine blue staining and immunofluorescence were used to check the effect of Rg1 on motor neuron and microglia. The expression of oxidative stress related protein Nrf2 and the miRNA were tested to investigate the mechanism of Rg1. We found that 20 mg·kg^-1·d^-1 Rg1 significantly postponed the disease onset and process, improved the motor syndrome, reduced the loss of motor neuron and inhibited the activation of microglia cells. Rg1 inhibited the aggregation of miR-153 in the spinal cord of ALS mice, which relieved the inhibition of Nrf2 and contributed to its up-regulation in the activation of HO-1 anti-oxidative signal pathway. Our study confirmed that Rg1 could protect ALS mice from oxidative damage through the up-regulation of miR-153/Nrf2/HO-1, which provides a theoretical foundation for Rg1 application to the ALS treatment.

This study was designed to explore proteins differentially expressed in HER2 positive gastric cancer N87 cells and N87/R cells with an acquired resistance to herceptin based on label-free quantitative proteomics. The extracted proteins were reduced and alkylated, then digested using filter aided sample preparation (FASP); peptides were separated via small manual reversed phase column, analyzed by LC-MS/MS, and identified with protein database 2.1 search engine. Proteins were quantified by intensity based quantification (IBQ) to search for differential proteins by comparison with relatively quantified proteins. The enrichment and network construction in gene ontology (GO) terms, genes-disease and Wikipathway of differential proteins were established through Web Gestalt. A total of 8 509 proteins were detected, among them, 7 163 proteins were further analyzed by bioinformatics, of which 110 proteins were up-regulated and 70 were down-regulated in N87/R cells. The differential proteins showed a significant difference in cellular component, biological process and molecular function in GO terms, respectively. Genes-disease network analysis indicated the association of these differential proteins with neoplasm metastasis, neoplasm invasiveness and inflammation, etc. Wikipathway enrichment analysis revealed the relevance of several signaling pathways to herceptin resistance, which included IL-2, MAPK/ERK, mTOR, aurora A, Ret, NF-κB, immune-regulatory and metabolic pathway. Western blot showed a significant increase of ERK1/2 activities in N87/R cells compared with N87 cells. Correspondingly, SCH772984, a MAPK/ERK inhibitor, preferentially reduced the viability of N87/R cells. Taken together, our data suggested that the MAPK/ERK signaling pathway is one of the key pathways that mediate herceptin resistance. This study provides the basic information for exploring the mechanisms of acquired resistance to herceptin in gastric cancer cells.

Study on the integrated pharmacokinetics/pharmacodynamics (PK/PD) model of rhubarb in rats with yang-deficiency constipation based on the principle of traditional Chinese medicine system. The rat model of yang-deficiency constipation was established using vinegar and ice water containing activated carbon. The blood samples with 0.5 mL were collected from orbital venous plexus at 0, 5, 10, 15, 30, 60, 120, 240, 480, 720, 1 440 min time points after oral administration of rhubarb decoction, and the dosage is equivalent to crude drug 2.5 g·kg^-1. The concentration of aloe-emodin, rhein, emodin and chrysophanol in rat plasma were determined by HPLC, and ELISA method was used to detect the activities of motilin (MTL), gastrin (GT), endothelin (ET) and vasoactive intestinal peptide (VIP) at different time points in serum. SPSS 21.0 software was used for analysis of component correlation and principal component, and WinNonlin 6.30 software was used to fit PK/PD model. Compared with the pharmacokinetic parameters of normal rats, in addition to emodin in the model rats showed characteristics of good absorption and slow to elimination; the content of MTL in model rats was significantly lower than that in normal rats. The composite values of the concentration and effect obtained by principal component analysis were connected by Sigmoid-E_max model. We established the integrated PK/PD model of rhubarb in treating yang-deficiency constipation to provide a new research direction for the material basis and mechanism of rhubarb treatment of yang deficiency constipation.

This study was designed to construct a "drug-core target-pathway" network of Erzhi Pill for hepatic injury treatment in an effort to explore the "multi-components, multi-targets, multi-pathways" mechanism. ADME/T calculation method was used to screen the active components of Erzhi Pill, and then predict the potential targets according to the reverse pharmacophore matching method. Biological information annotation databases (DAVID) was used to analyze the molecular function and biological process of the action targets. The Cytoscape software was used to construct the "ingredient-core target-pathway" network of Erzhi Pill for hepatic injury treatment. It was found that 39 major active ingredients of Erzhi Pill regulated 321 targets (HRAS, DCK, HSD17B1, UCK2, et al) and affected 51 pathways, such as insulin signaling pathway, FoxO signaling pathway, metabolic pathways and glycolysis/gluconeogenesis. The method revealed the action features of traditional Chinese medicine as multi-ingredients, multi-targets, multi-pathways, providing new clues for further basic study on the hepatic injury pharmacological mechanism of Erzhi Pill.

In this study, we used a mathematic-based modeling system to screen the cytokines that are sensitive to Zhuangguguanjie wan (ZGW)-induced idiosyncratic liver injury. The values of 27 cytokines were used as the data source in rat liver of lipopolysaccharide (LPS) + ZGW group. The alanine aminotransferase (ALT) activity value of liver function indexes was used as the outcome evaluation index of liver injury. Cytokines of ZGW-induced idiosyncratic liver injury were screened using Logistic regression, random forest method, LASSO Logistics regression and method of combining rule discovery algorithm with LASSO, and cytokines filtered out were revalued in THP1 macrophage. Susceptible cytokine combinations:interleukin-1β (IL-1β), epidermal growth factor (EGF) and interleukin-18 (IL-18) closely related to ZGW-induced idiosyncratic liver injury were obtained after preliminary screening analysis. The result of revalued in THP1 showed that the ethanolic extract of ZGW (EtZ) combined with IL-1β or IL-18 synergistically enhanced tumor necrosis factor-α (TNF-α) secretion in THP1 macrophage, and EtZ combined with IL-1β significantly enhanced interleukin-6 (IL-6) secretion in THP1 macrophage, but EtZ combined with EGF markedly inhibited IL-6 secretion in THP1 macrophage. The results suggest that the sensitive cytokines that can be characterized in the ZGW-induced idiosyncratic liver injury are IL-1β and IL-18, which provides a basis for screening the ZGW-induced idiosyncratic liver injury patients, and a new experimental evidence for clinical safety medication and risk prevention of ZGW.

Muscimol, cycloserin and ibotenic acid which are extracted from mushroom contain isoxazole pharmacophore structure. By using muscarinic structure as a model compound and molecular recombination method, we synthesized muscarinic analogues compounds 3-(1', 2'-di-O-isopropylidenedioxyethyl)-5-aryl-3a, 6a-dihydro-4, 6-dioxopyrrolino[3', 4'-d]isoxazoline derivatives through 1, 3-dipolar cycloaddition reaction. The structures of the target compounds were confirmed by UV-Vis, ¹H NMR, IR and elemental analysis. The drug activities of obtained compounds were screened in vitro. The pharmacophore in the structure is a potential non-covalent DNA binding, the compounds have anticancer activity and the leukocyte common antigen activity in a different extent. The preliminary results of in vitro anticancer test suggest that the inhibitory rates of compounds 3a-3o to Cdc25A phosphatase in cell division cycle ranged from 56.99%-99.94%; at the test concentration of 20 μg·mL^-1, 3f, 3h, 3i, 3m, 3o were no inhibition activity, and the rest of the compounds shows moderate to good excellent inhibition rate from 66.85% to 99.84%, even at the concentration as low as 5 μg·mL^-1. At the test concentration of 20 μg·mL^-1, except compound 3i, the rest compounds' inhibition activity of against leukocyte common antigen (LCA) CD45 protein tyrosine phosphatase A, are 63.08%-92.09%. These active compounds are potential inhibitors against Cdc25A and CD45 protein tyrosine phosphatase A, which have great application prospects in the treatment of cancers and Inflammatory and immune diseases.

Ten isoprenylated flavonoids were isolated from 95% ethanol extraction of Artocarpus heterophyllus, with a combination of various chromatographic approaches, including ODS, MCI, CHP-20 P, Sephadex LH-20 and high performance liquid chromatography. On the basis of physic-chemical characters and spectroscopic data analysis, these compounds were identified as artoheteroid E (1), cycloheterophyllin (2), artelastoxanthone (3), artoindonesianin Q (4), cudraflavone C (5), 8-(γ, γ-dimethylallyl)-5, 2', 4'-trihydroxy-7-methoxyflavone (6), kuwanon T (7), 6-(3-methylbut-2-enyl) apigenin (8), 5, 7, 2', 4'-tetrahydroxy-6-(3-methylbut-3-enyl) flavone (9), albanin A (10). Among them, compound 1 is a new one, while compounds 2-4 were isolated for the first time from the plant of Artocarpus heterophyllus. All isolated compounds were screened for their inhibitory abilities against cathepsin K. Of them, compounds 3-5, 7 and 10 showed inhibitory effects with the IC₅₀ values of 0.9, 1.6, 4.5, 24.5 and 63.5 μmol·L^-1, respectively.

This study was designed to establish a method to obtain the fingerprint chromatogram for the quantitative determination of Cordyceps sinensis in different sizes, a comparison of Cordyceps sinensis from five places was made to analyze its similarity and the content of main nucleosides (uridine, inoside, guanosine and adenosine). The assay was performed on a Waters XSelect HSS T3 C₁₈ (4.6 mm×250 mm, 5 μm), with a mobile phase consisting of water (A)-acetonitrile (B) at the flow rate of 0.6 mL·min^-1 (0-5 min, 0 B; 5-15 min, 0→10% B; 15-30 min, 10%→20% B; 30-33 min, 20%→50% B; 33-35 min, 50%→0 B; 35-40 min, 0 B). The detection wavelength was 260 nm and the column temperature was set at 30℃, and the injection volume was 5 μL. The results showed that there was no significant difference of the nucleosides in samples from the same place of the different sizes, but contents of the nucleosides variate a lot by production places. More data are required for further research. The method is proved for scientific and specific formulation of the standard in evaluation of circulated Cordyceps sinensis.

The study was designed to establish a 2D UPLC-QTOF method to extrapolate the structure of an unknown substance in carboplatin injection and its relationship with the excipient. By using phenyl-hexyl column (250 mm×4.6 mm, 5 μm) with mobile phase consisting of tetrabutylammonium sulfate buffer (pH 7.5) and acetonitrile in gradient elution mode, an unknown impurity in carboplatin injection was found and quantitatively determined. Then a 2D UPLC-QTOF, HSS T3 column (100 mm×2.1 mm, 1.7 μm) was employed to confirm the molecular weight and the structure of the unknown impurity (electrospray ionization source, positive ion mode, MS^E mode) with mobile phase consisting of 0.1% formic acid and acetonitrile. The relationship among impurities, API and excipient was investigated by accelerated stability test with ICP-MS/MS, ICP-AES. Results showed that disodium edetate in the formulation interacted with carboplatin producing an unknown impurity containing platin, and induced the increase of 1, 1-cyclobutanedicarboxylic acid. The research should be done on the rationality of the addition of disodium edetate in such injections containing heavy metals.

Naozhenning granule, which is composed of 11 herbal drugs, is mainly used in the treatment of concussion, cerebral post-traumatic syndrome. As the chemical composition of Naozhenning was complex, the ultrahigh performance liquid chromatography coupled with hybrid quadrupole-orbitrap mass spectrometry was applied to identify the chemical constituents rapidly in the Naozhenning granule. By analyzing the retention behaviors, accurate molecular weight, the fragmentation pathway, and comparison with the MS data from the standards and references, as well as the automatic identification by the Compound Discover 2.0, a total of 161 compounds were identified or tentatively deduced, and the sources of constituents were also confirmed. The compounds identified in Naozhenning granule included 9 iridoids, 8 butylphthalides, 26 flavonoids and flavonoid glycosides, 8 phenolic acids, 8 monoterpenoid glycosides and 9 alkaloids, as well as the common compounds in the herbal drugs, such as organic acids, amino acids, and sugars. The chemical composition of Naozhenning was studied for the first time, which provides a scientific basis for the quality control and document of effective materials of Naozhenning granule.

Due to the advantages of polymer micelles and the anticancer activity of doxorubicin (DOX), the polymer micelle of DOX is expected to be used for drug delivery in anticancer applications. As a biocompatible and biodegradable polymer, amphiphilic copolymer heparosan-adipic dihydrazide-vitamin E succinate (KV) can be self-assembled to form micelles with core-shell structure in aqueous phase. In this article, KV conjugates with two different degrees of substitution (DS) were synthesized to load DOX and were characterized by ¹H NMR. The size distribution, morphology, zeta potential and release behavior in vitro of the DOX-loaded micelles were studied. In vitro cytotoxicity was investigated by MTT assay against MGC80-3 and COS7 cells. The cellular uptake of the DOX-loaded micelles was observed by fluorescence microscopy and flow cytometry. The ¹H NMR spectra results confirmed the KV polymers were successfully conjugated and the degree of VES grafted on heparosan polysaccharide were 12% and 25%. Briefly, the micelles with two different DS were expressed as KV₁₂ and KV₂₅. The DOX-loaded micelles could resist serum adsorption because of the negative charge on the surface. The average particle size measured by dynamic light scattering (DLS) method was 140-150 nm and the TEM results indicated that the morphology of DOX-loaded micelles were spherical. The encapsulation efficiency and drug loading were 80% and 10%-15%, respectively. The DOX-loaded micelles had sustained release behavior and the cumulative release of DOX/KV₁₂ was slightly higher than DOX/KV₂₅. Moreover, the viabilities of cells which were co-incubated with blank micelles were greater than 90%. It is clear that the blank micelles almost non-toxic to both cells. The IC₅₀ of drug-loaded micelles against COS7 cells was much higher than that of MGC80-3 cells and the DOX/KV₁₂ exhibited greater cytotoxicity. The cellular uptake of DOX/KV on MGC80-3 was greater than COS7 cells. In this study, KV polymer micelles have a sustained drug release activity and have a good selectivity to tumor cells, so it would be a potential carrier in drug delivery.

The response surface method was used to evaluate the concentration range, property and degree of Scutellaria (HQ) and Coptis (HL) herb pair on improvement of glucose and lipid metabolism in this paper. Type 2 diabetes mellitus (T2DM) was induced in rats by low a dose of streptozotocin (STZ) in combination with high-fat diet feeding. The rats were administered with HQ-HL extract pairs at different ratio (1:0, 3:1, 2:1, 3:2, 1:1, 2:3, 1:2, 1:3, 0:1). To investigate the influence of herb pair extracts on glucose and lipid metabolism of T2DM rats, the multi index synthetic index method was adopted to integrate the indexes, then the response surface method was applied to analyze the integration results. The ratio of HQ and HL from 3:1 to 1:3 showed a synergistic effect (with the value between -0.2 and -0.8). Especially, when the proportion was between 3:2 and 1:3, herb pair extracts possessed stronger synergistic effects (value:-0.8). The high dose of HQ-HL with the ratio of 1:1 exhibited the strongest synergistic effect.

Flavonoids, especially chalcones such as hydroxysafflor yellow A and carthamin are the main active ingredients of safflower. To study the biosynthesis pathway of safflower flavonoids is of great significance for the quality control of safflower. Chalcone synthase (CHS) is an enzyme that plays an important role in regulation of the synthesis of flavonoids. However, for the time being, the role of CHS is not yet clear in the biosynthesis of safflower flavonoids. As a plant signaling regulator, JA/MeJA can activate CHS gene expression in plants. CtCHS1, one of the CHS genes in safflower, was elucidated in our previous work. In our continuous search for CtCHSs functions from this plant, other CHS genes CtCHS2 and CtCHS4 in safflower were examined. The floret was stimulated with methyl jasmonate (MeJA) and the transcriptome expression of CtCHS2 and CtCHS4 was analyzed by qRT-PCR at different time points of 0, 3, 6, and 12 h after stimulation. Further metabolites under stimulation by MeJA were analyzed by UHPLC/Q-TOF-MS. The results showed that the expression of CtCHS4 in response to MeJA significantly increased at 3 and 6 h, while the expression of CtCHS2 showed a trend of decrease after induction. Meanwhile, the accumulation of rutin, hydroxysafflor yellow A, D-phenylalanine, kaempferol-3-O-β-rutinoside and carthamin increased obviously. Especially, accumulation of hydroxysafflor yellow A was increased significantly at 3, 6 and 12 h after induction (P ≥ 0.05 or 0.01), but the change in kaempferol, kaempferol-3-O-β-D-glucoside, luteolin, quercetin-3-β-D-glucoside was not significant. The accumulation of hydroxysafflor yellow A and carthamin was positively correlated with the expression abundance of CtCHS4 with Pearson correlation analysis method (r ≥ 0.8). The data suggest that CtCHS4 may be a key gene for forming hydroxysafflor yellow A and carthamin and plays an important role in the accumulation of safflower chalcones. The CtCHS4-pMAL-C5X recombinant vector was successfully expressed in BL21 (DE3) Plys to express the product naringenin in vitro under the catalytic substrates p-coumaryol-COA and malonyl-CoA. The results of this study provide a new insight into synthetic genes involved in flavonoids biosynthetic pathway to elucidate the biosynthesis pathway of safflower chalcones.

Chinese pharmacopoeia stipulates that the content of liquiritin in licorice slices should be no less than 0.5%. However, there are lots of unqualified licorice slices in the herbal medicine markets. Due to the important role of functional gene polymorphism in secondary metabolism, this study attempts to analyze the influence of chalcone synthase (CHS) gene polymorphism on liquiritin biosynthesis and find out the unique haplotypes in licorice samples with high or low content of liquiritin, and to provide a basis for further analysis of molecular mechanism in flavonoid biosynthetic pathway. The contents of the 4 main flavonoids (liquiritin, isoliquiritin, liquiritigenin, isoliquiritigenin) in 60 licorice samples were assayed by HPLC and the results were analyzed by Spearman and χ² tests. The contents of the 4 main flavonoids were related to each other and obviously different in different original plants. They were highest in Glycyrrhiza uralensis samples and lowest in Glycyrriza inflate samples. Five G. uralensis samples with the highest liquiritin contents and five G. inflate samples with the lowest liquiritin contents were selected to clone the CHS cDNA sequences. 336 CHS cDNA sequences with a full length of 1 175 bp were obtained, 249 variable sites (141 missense mutation sites) were found, and 137 haplotypes were determined. 130 variable sites were found in the 336 CHS amino acid sequences and 102 types were determined. AA-3 is the major type of CHS in licorice, AA-35 is the special major type of CHS in the group with high flavonoids contents and AA-36 is the special major type of CHS in the group with low flavonoids contents. The mutation sites between AA-35 and AA-36 are I/V at 193 and V/T at 229. Discovery Studio 2.5 analysis of the three-dimensional structure of the CHS protein shows that the valine at site 229 of AA-35 is combined with malonyl-CoA. Homology analysis indicates that the homology of CHS among different species is low. This study is significant for identification of the unique haplotypes in licorices with high or low content of liquiritin and guiding the further molecular breeding of high-quantity licorice.