Drug-induced cardiotoxicity is recently a major concern. Cardiotoxicity is the leading cause of drug withdrawal from the market. Long-QT syndrome is one of the most important manifestations of cardiotoxicity. hERG potassium channel is an important target of drug-induced arrhythmia and antiarrhythmia drugs. Traditional Chinese medicine is a traditional medicine in China with a long history and a wide range of clinical use. However, the multi-organ toxicity caused by traditional Chinese medicine is still a problem to be solved. Some traditional Chinese medicines already in clinical use have been withdrawn from the market because of their potential cardiotoxicity or severe arrhythmias. The cardiac toxicity of more than 50 kinds of traditional Chinese medicines causing arrhythmia was reported, while more than 20 of them are induced by affecting on the hERG potassium channels. Therefore, finding out the mechanism of drug-induced long-QT syndrome and the regulatory target of drug intervention is the key research goal in today's medical field. In this paper, we summarized the mechanisms of long-QT syndrome induced by traditional Chinese medicine with Ikr/hERG potassium channel as the main target. It provides a theoretical basis for the rational use of related traditional Chinese medicine in clinical practice, the avoidance of cardiac toxicity and the development of regulatory targets for drug intervention.

Cardiotoxicity is one of the main causes of failure in new drug development or drug withdrawal from the market. However, current methods for evaluation of drug cardiotoxicity suffer the shortcomings such as low clinical relevance, low reproducibility and lack of high throughput screening capacity. Therefore, there is an urgent need for establishing more accurate and reliable methods for cardiotoxicity evaluation of drugs. As a new generation of drug cardiotoxicity evaluation, cardiac organs in culture retain the biological characteristics and functions of heart cells in the body, and can realistically and accurately respond to the effects of drugs. This article reviews recent progress of in vitro culture of cardiac organs and 3D-cell models, with focuses on application and development potential of cardiac organs for evaluation of cardiotoxicity of traditional Chinese medicine. The advantage and future prospective of such cell- and organ-based models for unique challenges in evaluation of cardiotoxicity of traditional Chinese medicine have been discussed.

Drug-induced cardiotoxicity is a serious concern in recent years, and acquired long QT syndrome (LQTS) is an important manifestation of cardiotoxicity. hERG gene encodes the α subunit of the rapidly activated delayed rectifier potassium channel (Ikr), which plays an important role in action potential phase 3 repolarization. Drug inhibition of Ikr/hERG channel leads to prolonged QT interval, accompanied by Tdp malignant arrhythmia, which can cause sudden death. We studied the effect of berberines on the hERG K⁺ channels after combination with rosuvastatin and glibenclamide, and evaluated the cardiac safety of these drugs in combination. Whole cell patch clamp technique was used to detect the effect of the combinations of these drugs on hERG current on HEK293 cells stably expressing hERG gene. The results showed that the inhibitory effects of berberine or dihydroberberberine combined with rosuvastatin on hERG current were higher than single drug (P < 0.05), but the combination had no effect on the kinetics of hERG channel. Berberine or dihydroberberberine combined with glibenclamide had higher inhibitory effects on hERG current than the application of single drug (P < 0.05) while the time constant of hERG channel inactivation was shortened after the combination (P < 0.05). In addition, the combination of berberine and glibenclamide inhibited hERG channel activation (P < 0.05). In conclusion, our results demonstrated that the combination of berberine with rosuvastatin or glibenclamide significantly inhibited hERG current and the inhibition effects were higher than the application alone. Therefore, when the two drugs that have inhibitory effects on the hERG channel are combined, the risk of inducing prolonged QT interval is significantly increased, and therefore reducing cardiac safety.

The aim of the present study was to investigate the effect of Sophoral flavones on proliferation of cardiac fibroblasts (CFb) induced by high glucose and its underlying mechanism. Cardiac fibroblasts were exposed to different concentration of D-glucose (15, 25 and 35 mmol·L^-1) at different time point (24, 48 and 72 h) in order to determine cell proliferation, and the model group was established by culturing CFb with 25 mmol·L^-1 D-glucose for 48 h. Sophoral flavones (12.5, 25 and 50 mg·L^-1) were employed for intervention. The cell viability was measured by MTT assay, and the levels of transforming growth factor-β₁ (TGF-β₁), matrix metalloproteinase-2 (MMP-2), collagen Ⅰ and collagen Ⅲ were measured by ELISA. In addition, flow cytometry was employed to detect the cell cycle; while the protein expression of prohibitin (PHB) was observed via immunocytochemistry and Western blot. This animal experiment had been approved by Jilin Medical University Experiment Animal Ethics Review Committee. The results showed that 25 mmol·L^-1 glucose could promote the proliferation of CFb; and the contents of TGF-β₁, MMP-2, collagen Ⅰ and collagen Ⅲ in the model group were higher than that of control (P < 0.05). The number of cells in S and G₂ phase increased under high glucose condition. In the model group, PHB translocation occurred at 6 h and protein expression decreased at 48 h (P < 0.01). Compared with the model group, 12.5-50 mg·L^-1 Sophoral flavones reduced the contents of TGF-β₁, MMP-2, collagen Ⅰ and collagen Ⅲ, increased the number of G₁ phase cells, and increased the expression of PHB protein at 48 h (P < 0.05), with no effect on the nuclear translocation of PHB. These results indicated that Sophoral flavones could prevent the proliferation of CFb induced by high glucose, the mechanism of which may be related to increasing the expression of PHB protein.

We evaluated the effects of Danggui-Chuanxiong (GX) herb pair with different proportions (1:0, 3:2, 1:1, 2:3, 0:1) and preparation methods (water extract W, alcohol extract A, and water-alcohol extracts WA) on vasoactive substances and endothelial cell adhesion molecules in the serum of acute blood stasis in rats. An acute blood stasis model was co-replicated by ice water bath and subcutaneous injection of epinephrine hydrochloride in rats. The expressions of vasoactive substances (arachidonic acid metabolites, coagulation-fibrin system index) and adhesion molecules in the serum were detected by enzyme linked immunosorbent assay method; the Spearman method was used to analyze the correlation of those detection indicators; the partial least squares-discriminant analysis and multi-attribute comprehensive index method were used to comprehensively evaluate the total effect of GX herb pair samples with different proportions and preparation methods on vasoactive substances and adhesion molecules. The experimental scheme was approved by the Animal Experimental Ethics Committee of the First Affiliated Hospital of Henan University of Chinese Medicine. The results showed that GX 1:1_WA had the strongest effect on the improvement of vasoactive substances and adhesion molecules in the serum of acute blood stasis in rats (the total effect value was 6.96). When extraction method was same, the overall effect of GX 1:1 had better effect than that of other proportions; when the proportion of GX was same, the total effects of GX_WA and GX_A were better than GX_W. The combination of Danggui and Chuanxiong can significantly improve the expressions of vasoactive substances and adhesion molecules in the serum of blood stasis in rats. But the action strength of GX herb pairs was different when the proportions and preparations of GX herb pair were different. These findings provide a basis for clinical rational application of GX herb pair, and lay the foundation for in-depth research on GX herb pair for treatment of blood stasis related diseases.

Long non-coding RNAs (LncRNAs), defined as transcripts which are hundreds of nucleotides with little or non-protein coding potential. Recently, LncRNAs have caught much more attentions, instead of considering as noises of genome transcripts, and indeed they have been found to play important roles associated with some biological processes, such as tumorigenesis, immunology dysfunction, metabolism adjustment, and so on. The incidence of chronic kidney disease (CKD) in different regions of the world is about 10% to 15%, with high growth rate and high unawareness, including the diabetic nephropathy, membranous nephropathy, etc. Previous publications also suggest that LncRNAs have a close relationship with the kidneys, and it may become new therapeutic targets or new biomarkers to diagnose diseases. In this review, we will summarize LncRNAs' functions with chronic kidney diseases, and discuss the prospects of the clinical applications of LncRNAs in the treatment of CKD treatment.

Provirus Integration in Maloney murine leukemia virus (PIM) represents a novel class of unique Ser/Thr kinase, which has been identified to be over-expressed in multiple hematological malignancies and some solid tumors, and the expression quantity correlates with malignant grade and poor prognosis in patients with cancer. PIM kinase plays important roles in regulation of cell proliferation and differentiation through the phosphorylation of its protein substrates, and it has become the emerging target for cancer treatment. A large number of highly active PIM kinase inhibitors have been reported by domestic and foreign research institutions, and the research progress will be summarized according to affiliations in this review.

Benzoxaborole is a series of compounds with five member ring and boron atom. Since the approval of crisaborole and tavaborole by FDA, benzoxaborole gained lots of research interests and become widely used in current drug discovery. Specially, benzoxaborole derivatives were found to exhibit anti-bacterial, anti-fungal, anti-protozoal, anti-tumor and anti-inflammatory activities. Here, we will review the properties of benzoxaborole, structure activity relationships as well as the recent progress in the biological activity of benzoxaborole derivatives.

Tumor, especially malignant tumor has become one of the major diseases, a serious threat to the health of people around the world. Modern clinical practice shows that the natural active products extracted from traditional Chinese medicine, marine medicine and other natural drugs, such as terpenes, alkaloids, polysaccharides, volatile oils and peptides, can effectively inhibit the growth of tumor cells. In this paper, the active components of natural antitumor products in recent years were summarized and their related mechanism was elucidated, so as to provide theoretical basis for the further development of natural antitumor drugs.

Chiral amino acid analysis is a sensitive, efficient and economical method for controlling racemic peptide impurities, especially for synthetic polypeptide drugs with complex composition of amino acids. Unexpected amino acid enantiomers in racemic peptides can be measured by chiral amino acid analysis coupled with mass spectrometry. The position of amino acid isomerization in the peptide segment can be accurately mapped by mass spectrometry, which lays a solid foundation for screening of racemic peptide impurities and rapid identification or quantification of trace racemic peptide impurities. Combination of the two techniques is vital for quality control of the synthetic polypeptide drugs and for research of polypeptide drugs based on chemical synthesis. The strategies of peptide hydrolysis have been summarized in this review. The latest chiral amino acid analysis based on mass spectrometry is briefly reviewed. Based on our knowledge, we have pointed to the direction of research and control of racemic peptide impurities in synthetic polypeptide drugs.

The in vivo fate is a crucial factor that governs the successful translation of nanoformulations. However, one of the current biggest challenges is with the real-time monitoring of the body of the nanoparticles themselves. Conventional radioactive or fluorescent probes give signals even after they are disassociated from the particle matrix, generating interference to bioimaging and leading to misjudgment of results. Environment-responsive fluorescent dyes are regarded as promising tools due to signal switching in response to the changes in the environment. Currently, there are three categories of dyes in bioimaging of nanoparticles based on Förster resonance energy transfer (FRET), aggregation-induced emission (AIE) and aggregation-caused quenching (ACQ). They have similar characteristics that strong fluorescence is emitted when they are embedded in the matrix of nanocarriers, whereas the fluorescence quenches upon release from the matrix due to dissociation of nanocarriers. The fluorescence switching reflects the existing status of the nanocarriers and therefore helps to interpret the in vivo behaviors. FRET and AIE probes have been widely used in elucidating the interactions between nanoparticles and cell models. However, they show intrinsic defects in studying in vivo fate of nanoparticles. ACQ-based dyes are sensitive to water, a universal factor in the biological environment. Therefore, with the help of bioimaging equipment, the in vivo trafficking process of nanoparticles can be unraveled. This review article tends to provide an overview on the rationale, pros and cons and applications of the three categories of environment-responsive fluorescent dyes in the investigation of the in vivo fate of nanocarriers.

We studied the protective effect and mechanism of isorhamnetin (ISO) on 1-methyl-4-phenylpyridiniumion (MPP⁺)-induced SH-SY5Y cells injury. MPP⁺-induced SH-SY5Y cell injury model was established, and cell viability was measured by MTT and LDH methods. The activity of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in cells were determined to investigate the level of oxidative stress. DCFH-DA and MitoSOX fluorescence probes were used to detect the levels of intracellular reactive oxygen species (ROS) and mitochondria superoxide, respectively. JC-1 fluorescence probe was used to detect the changes of mitochondrial membrane potential. Western blot and immunofluorescence methods were used to determine the expressions of Sirt1 and PGC-1 proteins, as well as the expression levels of apoptosis-related proteins Bax and Bcl-2. MPP⁺ at the dose of 500 μmol·L^-1 significantly reduced SH-SY5Y cells viability to 52.46% and increased LDH release to 417.63%. ISO at 5 and 15 μmol·L^-1 significantly increased the expression of Sirt1 and PGC-1α, inhibited LDH release, reduced intracellular ROS and mitochondria superoxide, inhibited the decline of mitochondrial membrane potential and increased cell viability to 61.61% and 67.55%. In addition, ISO could downregulate the expression of Bax and upregulate the expression of Bcl-2 to reduce cell apoptosis. ISO-mediated inhibition of apoptosis could be reversed by Sirt1 specific inhibitor Sirtinol. Through activating Sirt1/PGC-1α signaling pathway, ISO could reduce oxidative stress injury and inhibit cell apoptosis to protect cells from MPP⁺ injury.

Seven main components in eleutheroside were used as research objects, and the mechanism of action of total eleutheroside for treatment of diabetes mellitus type 2 was investigated by network pharmacology. The SwissTargetPrediction, GeneCard, and String platforms were used to predict the 35 potential targets of these 7 components that are related to diabetes mellitus type 2. Then we used cytoscape 3.6.1 to build a " component-target" network map and used the Networkanalyzer tool for topology analysis. Gene ontology (GO) enrichment analysis and KEGG pathway enrichment analysis were performed on the DAVID6.8 platform, and the " component-target-path" network map was constructed based on the enrichment results. Those components mainly used in diabetes mellitus type 2 were screened as core components, and the core components were docked with key disease target proteins to verify the potential mechanism of the total eleutheroside. After screening, 8 important pathways associated with diabetes mellitus type 2 were identified. This study showed that eleutheroside A, eleutheroside D, eleutheroside E and sesamin played key roles in insulin resistance, apoptosis and inflammation pathways. The total eleutheroside may ameliorate type 2 diabetes mainly through regulating signal transducer and activator of transcription factors (STATs), non-receptor protein tyrosine phosphatase (PTPN) 1, PTPN2, c-Jun N-terminal kinase (JNK), and p38 mitogen activate protein kinase. These components worked together through multiple signaling pathway. Based on our data, eleutheroside is proposed as a novel therapeutic strategy for treatment of type 2 diabetes.

Platelet adhesion is a key process in thrombosis. Anti-platelet adhesion effect of some Chinese medicines for promoting blood circulation and removing blood stasis (PBCRBS) has been reported, but their relative efficacies as a whole and specific targets remained unclear. This paper combined activity screening, drug compatibility analysis, pathway clustering, target prediction, and molecular docking to explore the mechanism of anti-platelet adhesion by PBCRBS Chinese medicine. Screening the activity of anti-platelet adhesion of 58 commercially available PBCRBS Chinese patent medicines showed that about 50.0% significantly inhibit ADP-induced platelet adhesion in vitro, and about 96.6% significantly inhibit thrombin-induced platelet adhesion in vitro. The animal experiment involved was approved by the Animal Ethics Committee of Tianjin International Biomedical Research Institute. Combined with the auxiliary platform for TCM (V2.0) inheritance showed that the compatibility of Danshen-Chuanxiong was used most frequently among the top 20 active proprietary Chinese patent medicines. IPA network analysis revealed that IL-1, APP and CCL2 might be the key targets for anti-platelet adhesion function of Danshen-Chuanxiong against atherosclerosis, neuroinflammation and chemokine signaling pathways as the main mechanisms. Molecular docking analysis confirmed the interaction between one of the active compounds shared by Danshen and Chuanxiong, i.e. chlorogenic acid, with its target CCL2. This study provides TCM theory guidance and experimental support for targeting platelet adhesion in anti-thrombosis therapy by Chinese medicine for promoting blood circulation and removing blood stasis.

Paeoniflorin (PAE), the major active compounds of Chinese herbs Radix Paeoniae Alba and Chinese patent drug " Total Glucosides of Paeony Capsules" , which is effective in the treatment of rheumatoid arthritis (RA), exerted multi-pharmacological activities, such as anti-inflammatory, immune-regulatory, etc. However, its potential action mechanisms remain unclear. Herein, we predicted the putative targets of Radix Paeoniae Alba and constructed an interaction network of putative targets of Radix Paeoniae Alba and known RA-related genes. A list of key putative targets was identified by calculating their topological features (degree, node betweenness and closeness) in the above pharmacological network. Importantly, pathway enrichment analysis revealed that these key putative targets were significantly enriched in several RA-related pathways, including cartilage damage-related IL1B-TNF-TLR2-JUN-MMP1-MMP3 signaling pathway. Further molecular docking simulation showed that PAE, the major active compounds of Radix Paeoniae Alba, has strong binding affinity with MMP1 and MMP3 proteins. Next, in vivo experiments based on the adjuvant-induced arthritis (AIA) animal models showed that PAE significantly alleviated the disease severity and the syndromes of severe redness or swelling in hind limbs of AIA rats, including decreasing the arthritis score, the diameter of the limbs, and elevating body weight and pain thresholds (all P < 0.05). ELISA assay indicated that PAE obviously suppressed the abnormal up-regulation of serum inflammatory factors including IL-1β, TNF-α, IL-6, IL-17 and IFN-γ in AIA rats (all P < 0.001). Western blot analysis found that PAE simultaneously modulated the abnormal up-regulation of MMP1 and MMP3 proteins in the ankle tissues of AIA rats (all P < 0.001) (all procedures in the current study were performed in accordance with the ethical standards of the Center for Laboratory Animal Care, China Academy of Chinese Medical Sciences). In conclusion, PAE alleviated the cartilage damage and disease severity in the progressive process of RA via regulating the IL1B-TNF-TLR2-JUN-MMP1-MMP3 pathway. This study provided the theoretical basis of the PAE for its immune-regulatory effects, and as well provided references for the action mechanism study of extract compounds of Chinese herbs.

The Lewis lung carcinoma (LLC) metastatic mouse model was used to investigate the effects of gefitinib and Sijunzi Tang (SJZ) on pre-metastatic niche. The experimental protocol was approved by the Ethics Committee which belongs to Cancer Hospital, Chinese Academy of Medical Sciences. To generate spontaneous lung metastatic models, 1×10⁶ luciferase-labeled LLC cells were injected subcutaneously in the shaved right flank of mice. One day after LLC inoculation, the mice were randomly divided into model (saline), gefitinib (50 mg·kg^-1) treatment, SJZ treatment (25.74 g·kg^-1), and co-treatment gefitinib with SJZ groups, with intragastrical administration. After 14 days of continuous administration, tumor size was detected by IVIS^® Spectrum system. The number of monocytes and neutrophils and the expression levels of chemokine receptors (CXCR1, CCR2) and carcinogenic gene (c-Kit), in peripheral blood, spleen and lung tissues of mice were determined by flow cytometry. The contents of interleukin-IL-1α (IL-1α) and interleukin-6 (IL-6) were detected by the enzyme linked immunosorbent assay (ELISA). After 21 days of treatment, tumors were surgically removed, weighed and the tumor volume was measured with vernier caliper and the antitumor effect of co-administration was evaluated. After 45 days of administration, the survival of mice was recorded. The results of flow cytometry showed that the percentage of neutrophils in gefitinib group, SJZ group, and co-treatment group was significantly decreased in the lung tissue compared to the model group (P < 0.05 or P < 0.01), but there was no significant difference between three treatment groups (P > 0.05). In the mouse peripheral blood and lung tissue, compared with the model group, the expression levels of CXCR1, CCR2 and c-Kit on the surface of neutrophils and monocytes in SJZ group and co-treatment group decreased or decreased significantly (P < 0.01 or P < 0.05). However, there was a significant increase in the expression level of c-Kit on the surface of monocytes (P < 0.05). In the mouse spleen tissue, the expression levels of CXCR1, CCR2 and c-Kit in the gefitinib group increased significantly (P < 0.05), while decreased significantly in SJZ or co-treatment group (P < 0.05). The results of ELISA showed that the content of IL-1α in SJZ group decreased significantly in the plasma of the mice compared with the model group (P < 0.01) and the content of IL-6 in co-treatment group decreased significantly (P < 0.05). Compared with the gefitinib group, the content of IL-1 in the co-treatment group decreased significantly (P < 0.05). In the tumor tissues of mice, compared with the model group, the content of IL-1α in the co-treatment group decreased significantly (P < 0.05). Furthermore, the content of IL-1α in co-administrated group and IL-6 in SJZ or co-treatment group decreased significantly compared with the gefitinib group (P < 0.05). After 21 days of continuous administration, the tumor inhibition rates of gefitinib group, SJZ group and co-administrated group were 45.7%, 38.4%, and 84.8%, respectively. After 45 days of administration, the survival rate of the model group was 0%, whereas the gefitinib, SJZ or co-treatment group has a survival rate of 40%, 60%, or 60%, respectively. In summary, our study illustrated that Sijunzi Tang could improve the anti-tumor effect of gefitinib by regulating pre-metastatic niche.

Cichorium glandulosum has been used to treat non-alcoholic fatty liver disease (NAFLD) and type 2 diabetes mellitus (T2DM) in Uyghur folk medicine. The mechanism of Cichorium glandulosum (CG) on type 2 diabetes mellitus accompanied with non-alcoholic fatty liver disease (T2DM-NAFLD) remains unclear. The effect of CG extraction on T2DM-NAFLD was determined in animal experiments here (all the experiments here were approved by the Animal Care Committee of the First Affiliated Hospital of the Medical College, Shihezi University). The mechanism of CG for treatment of T2DM-NAFLD was predicted and verified based on systems pharmacology. Based on the active compounds of CG on T2DM-NAFLD, T2DM and NAFLD-related targets, pathways and diseases were screened and predicted. Active compounds-targets, compounds-targets-pathways and compounds-targets-diseases were constructed and analyzed. The results of animal experiments showed that CG extraction can reduce the levels of blood glucose and blood lipid in T2DM-NAFLD rats. In addition, it can improve the glucose tolerance and relieve liver injury. Total 29 active compounds and 198 targets were screened by systems pharmacology, of which 106 targets were involved in T2DM, 88 were involved in NAFLD, and 56 targets were common between T2DM and NAFLD, mainly related to insulin resistance and inflammation. These 198 targets include those in metabolic pathways, calcium pathway, PI3K/Akt pathway, cAMP pathway, and MAPK pathway. Our study confirmed that CG can be potential phytomedicine for treatment of T2DM-NAFLD. This work provides a reference for studying the treatment of multiple diseases using multiple-targets phytomedicine in systems pharmacology.

Polymyxin B and polymyxin E (colistin) are increasingly used as last-resort drugs for treatment of infections caused by multidrug-resistant gram-negative pathogens. Unfortunately, the application was limited due to the serious side effects, especially nephrotoxicity. Very recently, the need for developing more tolerated and more effective polymyxin analogues has grown. This study details the design, synthesis, and evaluation of two classes of polymyxin B analogues with varying hydrophobicity and bulkiness at the N-terminal fatty acyl chain or position 6 amino acid. 20 polymyxin B analogues were synthesized and the chemical structures of the analogues were confirmed by HR-MS and ¹H NMR spectra. Compounds 7e (MIC:0.5-4 μg·mL^-1) and 7l (MIC:0.25-2 μg·mL^-1) showed similar or better antimicrobial activity against both susceptible and resistant strains of Escherichia coli, Klebsiella pneumoniae, Acinetobacter baumannii, and Pseudomonas aeruginosa compared to polymyxin B (MIC:0.5-2 μg·mL^-1). Besides, compound 7l (CC₅₀:217.1±13.2 μg·mL^-1) displayed noticeably decreased renal cytotoxicity compared to polymyxin B (CC₅₀:120.3±6.0 μg·mL^-1). This work establishes the base of further study on the structure-activity relationship of polymyxin B.

Indoleamine 2, 3-dioxygenase 1 (IDO1) is a key enzyme in the human tryptophan metabolism pathway, which can mediate tumor immune response. An IDO1 inhibitor would be a potential cancer immunotherapy drug. Based on the recently reported crystal of an IDO1 protein-inhibitor complex (PDBID:6AZV), the structure of reported inhibitor, and by analyzing the interaction mode between the inhibitor and IDO1, new inhibitor molecules were designed and synthesized. All structures were confirmed by spectral data. Preliminary activity studies showed that compounds containing an azabiphenyl tetrazole structure (B1 and B2) and biphenyl compounds containing a sulfonamide structure (D1, D2 and D3) had excellent inhibitory activity of IDO1 at the enzyme and cell level, and were comparable or even better than the control drug INCB24360.

The chemical constituents from Brassica rapa were identified by various chromatographic techniques including silica gel, reversed-phase silica gel, macroporous resin and Sephadex LH-20 column chromatography. Eight compounds were isolated from this plant. The isolated compounds were elucidated by physicochemical properties and spectroscopic methods, including extensive 1D, 2D-NMR, and HR-ESI-MS techniques. Compounds 1, 2 are new triterpenoids and 3-7 were isolated for the first time from Brassica rapa. The cytotoxic effect of compounds 1 and 2 were tested by MTT assay against five cancer cell lines. The result showed that all compounds exhibit growth inhibition for the cancer lines. Compound 1 has an IC₅₀ value of 5.87 μmol·L^-1 for growth inhibition of leukemia cell line HL-60, and IC₅₀ value for compound 2 was 10.32 μmol·L^-1.

The chemical constituents of the aerial parts of Lespedeza cuneata (Dum. Cour.) G. Don were investigated using chromatographic techniques, such as silica gel, reversed phase MPLC and preparative HPLC. Five compounds were isolated and their structures were elucidated by spectroscopic data and physicochemical properties, which were identified as 7-O-glucosyllaburnetin (1), kaempferol-3-O-β-D-galactopyranoside (2), kaempferol-3-O-α-L-rhamnoside (3), vitexin (4), and isovitexin (5). Among those, compound 1 is a new compound, compounds 2-3 were isolated from this plant for the first time. Compounds 1-5 were tested for their anti-ulcerative colitis activity by dual luciferase report gene assay targeting xbp1. Compared with control group, compound 1 showed a certain activity on activating the transcription of xbp1, with its relative activating ratio being 1.80 times.

An HPLC fingerprint and multi-component determination method of Leonurus japonicus was established for comprehensive evaluation and quality control of Leonurus japonicus. The sample was incubated in 70% ethanol in a water bath for 2 h, and the extract was analyzed by HPLC using Kromasil C18 column (250 mm×4.6 mm, 5 μm). The mobile phase consisted of acetonitrile-0.1% formic acid with gradient elution. The flow rate was 1.0 mL·min^-1. The temperature of column was 30℃. The detection wavelength was 280 nm. HPLC fingerprint of characteristic components of Leonurus japonicus was established. There were 12 common peaks among 25 batches of samples, and 5 of them were identified and determined. Syringic acid, leonurine hydrochloride, rutin, hyperoside or isoquercitrin showed a good linearity in the ranges of 0.426 1-85.22 ng (r=0.999 9), 7.948-1 590 ng (r=0.999 3), 10.20-2 040 ng (r=1.000 0), 2.018-403.6 ng (r=0.999 9), or 8.704-1 741 ng (r=0.999 9), respectively. The average recoveries were 99.0%, 97.6%, 97.4%, 96.9% and 98.5% with RSD of 1.1%, 1.8%, 1.4%, 1.5% and 1.3%, respectively. The HPLC characteristic fingerprint of Leonurus japonicus was specific, and this method can simultaneously determine the content of 5 components.

Vinpocetine (VP) has been widely used to treat cerebrovascular disorders and nerve injury. Borneol (BN), as an important traditional Chinese medicine, is commonly used to promote the absorption and distribution of central nervous system drugs. In this work, a LC-MS/MS method was developed to determine the level of VP in rat plasma and tissues, and to evaluate the effect of co-administration of BN with VP by oral gavage on the absorption and tissue distribution of VP in rats. Rats were divided into VP (10 mg·kg^-1), VP (10 mg·kg^-1) + BN (75 mg·kg^-1) and VP (10 mg·kg^-1) + BN (150 mg·kg^-1) groups for pharmacokinetic study, and divided into VP (10 mg·kg^-1) and VP (10 mg·kg^-1) + BN (150 mg·kg^-1) groups for tissue distribution study. The animal experiment was approved by Ethics Committee of Hubei University, and complied with the guideline for caring and using of laboratory animals. Compared to VP group, the AUC_0-∞, MRT_0-∞ and t_1/2z of VP + BN (150 mg·kg^-1) group increased significantly, by 1.98-, 1.22- and 1.42-fold respectively, and the exposure in plasma, liver, kidney and brain increased by 2-, 1.5-, 1.5- and 1.3-fold respectively. The pharmacokinetic results suggested that co-administration of BN with VP is beneficial for overcoming the undesirable pharmacokinetic characteristics of VP, such as short residence time, low oral bioavailability and brain exposure in clinical usage.

The preparation of polymorphic forms of rivaroxaban was carried out using a recrystallization method based on that for crystal form-Ⅰ. Preparation methods were developed for two crystal forms-Ⅱ (medicinal crystal form) and five crystal forms-Ⅳ and the crystals were then characterized. The crystalline form was identified by applying modern analytical means including X-ray powder diffraction (PXRD), differential scanning calorimetry (DSC), element analysis (EA), mass spectrometry (MS), and infrared spectrum (IR), and the morphology of different crystal forms was observed by scanning electron microscopy (SEM). The results show that the PXRD and DSC characteristics of prepared crystal forms-Ⅱ and Ⅳ are consistent with those described in patents at home and abroad, and the test results with EA, MS and IR are in accordance with the chemical structure of rivaroxaban. The crystal form-Ⅰ is lamellar, the crystal form-Ⅱ is linear and crystalline form-Ⅳ is striped as determined by SEM. In summary, the methods for preparing crystal form-Ⅱ and form-Ⅳ are reliable, the required reagents are easily available, the experimental conditions are easy to implement and the preparation process is simple. Our study provides a new reference for the production and application of rivaroxaban.

We determined a component-target-disease network for Carthamus tinctorius L. and the key compounds, identified by topological analysis, were related to vasculitis, coronary heart and cerebrovascular disease. Based on these compounds, the chromatographic fingerprint of Carthamus tinctorius L. was established. Firstly, 132 compounds were obtained from TCMID and TCMSP databases. Their targets were predicted in the PharmMapp and HemMapper databases. CardioGenBase, Therapeutic Target Database and DisGeNET databases were used to collect targets of vasculitis, coronary heart disease and cerebrovascular disease. The corresponding relationships between component and target protein were established by mapping. Finally, the " component-target-disease" network was built with Cytoscape software. The core network and key nodes were analyzed with the Cytohubba plug-in. The results showed that the 24 key compounds were alpha-tocopherol, adenosine, quinone chalcone pigments such as hydroxysafflor yellow A, safflower yellow, quercetin, kaempferol and other flavonoids, organic acids such as stearic acid, linolenic acid, coumaric acid and cinnamic acid. This resulting chromatographic fingerprint of Carthamus tinctorius L. showed good consistency, and the core chemical compounds obtained by topological analysis of the network of " component-target-disease" , could be used as quality control markers. Our research provides a new approach for the identification of quality control indicators in Chinese medicinal materials.

The morphological identification, chemical component analysis, and DNA barcode determination were investigated on Genus Mentha (including Mentha haplocalyx Briq., Mentha piperita Linn., Mentha spicata Linn. and Mentha cultivated) in order to reveal the origin of Herba Mentha as a drug, and ensure the accuracy in clinic application. The morphological characters, chemical composition analysis by GC-MS/MS and DNA content measure by polymerase chain reaction (PCR) were reported in this study for inter- or intra- species divergence. Based on the morphology, axillary verticillasters was recognized as the typical character for Mentha haplocalyx Briq. Carvone was used as an index component for chemical composition analysis of Mentha spicata Linn. Gene clustering analysis divided 22 batches of samples into two molecular groups. Mentha haplocalyx Briq. is distinguishably different from Mentha spicata Linn. Mentha piperita Linn. and other cultivated plants were distributed between these two species. The results obtained by morphological identification, chemical composition analysis, and DNA barcode determination show good correlations, but each identification method has its limit. In view of the fact that hybridization of the plants in Genus Mentha is common, identification relying on only one method is not recommended.

The methods for determination of freezing point include cooling curve cryoscopy and air humidity cryoscopy. These methods are usually influenced by many factors, such as instrumentation, environment and operators. Despite the numerous experimental methods, precise freezing point values are challenging to obtain due to time-consuming procedures, limited sample size and extensive manual work. In this study, a semi-empirical hydration model (SEHM) was developed to calculate freezing point of NaH₂PO₄-K₂HPO₄ mixed electrolyte solution. According to SEHM, the water activity of mixed electrolyte solution was calculated by molality of solutes. Then the freezing point of solution was calculated. The calculated results were compared with those obtained by cooling curve cryoscopy and air humidity cryoscopy. The results indicate that the SEHM calculations are comparable to the measurements of cooling curve cryoscopy and air humidity cryoscopy.

Bioadhesive preparation can be attached to specific sites to control drug release rate, increase drug concentration and increase efficacy, which is based on natural or synthetic polymer material. In this paper, based on the physical properties of wet mass, a method for screening adhesion formulation was proposed, which was different from conventional way of screening optimal formulation, and astragalosides loaded bioadhesive pellets were prepared by extrusion-spheronization method (extrusion speed 30 r·min^-1, spheronization speed 808 r·min^-1, spheronization time 7.5 min) based on this formulation screening method, small living animal imaging technology and mucin from porcine stomach model were used to evaluate the in vivo and in vitro adhesiveness behaviour of the pellets. According to the relationship between the physical properties of wet mass and the formability and adhesiveness of bioadhesive pellets, five key physical properties hardness (Ha), adhesiveness (Ad), springiness (Sp), cohesiveness (Co), chewiness (Ch) were selected as the index of screening optimal formulation, therefore a comprehensive evaluation model was established, which based on principal component analysis, to did digital ranking for these proposed adhesion formulation, the optimal formulation was determined:microcrystalline cellulose:(chitosan:Carbomer 940=2:1), the adhesive material dosage accounted for 20% of the excipient dosage, and the ratio of drugs to excipients was 1:4. All animal experiments have been approved by Ethics Committee of Shanghai University of Traditional Chinese Medicine. The in vivo and in vitro adhesive evaluation results showed the pellets had a clear advantage in intestinal adhesion over normal pellets, its also proved the scientificity and reliability of the method of screening bioadhesive formulation.

The structural composition of the surface fungal community of commercially Platycladi semen was analyzed to reveal the surface fungal biodiversity and structural differences. Platycladi semen was collected from Henan, Shandong and Hong Kong, their DNA was extracted, ITS fragments in DNA were amplified by PCR. Miseq was sequenced on Illumina Hiseq 2500 platform after the PCR products were qualified for quality inspection. The sequence OTU cluster was obtained and bioinformatics analysis was carried out. Microbial communities were not observed in the eyes of the Platycladi semen in the three regions. Sequencing results showed that the surface microbial community had high biodiversity, but there were significant differences in species composition. Seven samples o Platycladi semen obtained 345 947 valid sequences, which were divided into 267 OTUs, 3 phylums. 18 classes, 40 orders, 82 families, 120 genus, 191 species fungi. At the genus level, Aspergillus is the dominant species, accounting for the highest proportion, reaching (93.36 ±6.01)%. Seven samples were contaminated by Aspergillus flavus, and the pollution levels were 14.58%, 15.98%, 17.64%, 16.44%, 0.97%, 23.39% and 18.86%. Except sample No. 5, Aspergillus cibarius was the most abundant, the other six samples were Aspergillus niger, Aspergillus fumigatus and Aspergillus flavus as the core microflora. By analyzing the diversity of fungi distribution in different habitats, we can fully understand the fungi on the surface of Platycladi semen, lay a foundation for early risk warning of Aspergillus flavus contamination and its aflatoxin contamination, and provide a theoretical basis for the quality and safety of Platycladi semen.

In order to explore the chloroplast genome characteristics of Sophora flavescens and the phylogenetic relationship of the genus, this study used high-throughput sequencing technology to sequence and functionally annotate the chloroplast genome of Sophora flavescens. The results showed that the full length 154 165 bp of Sophora flavescens chloroplast genome showed a typical four-stage structure. The chloroplast genome contains 123 genes, including 77 protein-coding genes, 38 tRNA genes and 8 rRNA genes. Sequence analysis revealed 104 SSR loci, most of which consisted of A and T. In addition, the chloroplast genome codon preference is weak, and the coding region is biased towards the use of A and T bases. A comparative analysis of two different regions of Sophora flavescens chloroplast genome revealed four differential genes. Based on the maximum likelihood method (ML) for phylogenetic analysis of Sophora flavescens and 16 other leguminous, it was found that the relationship between Sophora flavescens and the genus Sophora alopecuroides is the closest. This study provides an important theoretical basis for the genetic variation, breeding and phylogenetic analysis of Sophora flavescens, and has certain reference value.

Sequence analysis of DNA, mRNA and protein is an essential component of biologics or bioprocess development. Analysis of sequences at the DNA, mRNA, and protein levels after the transfer of the gene of interest into a host cell is an important part of quality control. This article reviews the application of new technologies such as next-generation sequencing and LC-MS/MS in biological drug development such as monoclonal antibodies. These techniques have different requirements in term of cost, handling time and expertise. Selecting an appropriate technique with a sound rationale at different stages of drug development will add to the success rate of research and development, and ensure product quality, thus ensuring the clinical efficacy and safety.

Recently, biosimilar antibodies have become a mainstream component of the biopharmaceutical industry in China. The principle requirements for the development and evaluation of biosimilars are based on proving similarity in product quality (analytical similarity) between a proposed biosimilar candidate and the originator reference drug. However, because the quality of a reference drug often varies during the life cycle and not all reference drug samples are able to collected by a biosimilar sponsor, it has not been practical to accurately determine the critical quality attributes as well as an accurate control range through the characterization of the limited number of reference drug lots that are typically collected. Therefore, the development and evaluation of biosimilars has been challenging both for industry and regulatory agencies. In this article, The Chemistry, Manufacturing and Control (CMC) dossier of the rituximab originator company and the dossiers of 18 biosimilar companieswere retrospectively analyzed. Furthermore, the assessment criteria to determine quality similarity of rituximab biosimilar candidates have been proposed, which criteria have been used by reviewing the physicochemical and biological properties data obtained from 123 lots of the reference drug. Moreover, some case studies have been provided that illustrate the application of the proposed analytical similarity criteria in the practice of drug evaluation.

The marketing authorization application is a milestone of drug life cycle, which indicates a candidate has potential to become a commercial drug. As of now, there are only 12 domestic therapeutic antibodies approved in China. The chemistry, manufacturing and controls (CMC) development and evaluation of monoclonal antibody were more challenging for both industry and authority agency. As the result of domestic biopharmaceutical industry development and implement of priority review system, the marketing authorization application of domestic antibody biosimilar and imported antibodies had dramatic increased in recent years. Thus, the CMC evaluation of monoclonal antibody become the important task of biological product's marketing authorization registration management. In the article, the CMC regulatory considerations for marketing authorization application based on author's review experience was proposed, in order to accelerate development and registration of commercial antibody in China.