The three polymorphs (Ⅰ, Ⅱ, Ⅲ) and one amorphous (Ⅳ) form of imatinib-oxalate new salt were prepared and characterized by powder X-ray diffraction, differential scanning calorimetry, thermogravimetric analysis and infrared spectroscopy, and transformation rules among the four salts were analyzed. The solubility of polymorph Ⅱ was investigated by high performance liquid chromatography. The results show that polymorph Ⅱ was stable but polymorph Ⅰ, Ⅲ and amorphous Ⅳ were unstable at room temperature. Polymorph I transformed into polymorph Ⅱ at room temperature. At relative humidity 90%, polymorph Ⅰ, Ⅱ and amorphous Ⅳ transformed into polymorph Ⅲ, but polymorph Ⅲ transformed into amorphous Ⅳ at 70 ℃. The solubility of imatinib was significantly improved after being salted with oxalic acid, and the equilibrium solubility of polymorph Ⅱ in pH 6.8 buffer solution and pure water was increased by 4.1 and 21.2 fold, respectively, as compared to bulk drug. This research provides guidance for the development and quality control of a new salt form of imatinib.

The plant-specific WRKY transcription factor family is involved in the regulation of the response of plants to various environmental factors and biological stress. To study the role of the BcWRKY70 gene in Bupleurum chinense DC., we cloned the open reading frame (ORF) sequence of the BcWRKY70 gene based on the transcriptome database. Bioinformatics analysis of BcWRKY70 and promoter region analysis was carried out, and real-time qRT-PCR was used to detect the expression pattern of BcWRKY70 in different tissues and under the different hormone treatments. The results show that the BcWRKY70 ORF is 948 bp in length, encoding 316 amino acids with a typical WRKY domain. Phylogenetic analysis showed that BcWRKY70 is closely related to GhWRKY70. Its promoter region contains cis-elements that presumably respond to environmental factors such as drought, methyl jasmonic acid (MeJA), and abscisic acid (ABA). qRT-PCR analyses showed that BcWRKY70 is most highly expressed in roots and can quickly respond to MeJA and ABA induction. This study provides a basis for further research on the molecular mechanism of BcWRKY70 in B. chinense DC. resistance.

A model for quality evaluation was developed by determination of multi-index components in Scutellaria baicalensis Georgi. Eight flavonoids in Scutellaria baicalensis Georgi were quantified by high performance liquid chromatography (HPLC), and a quantitative analysis of multi-components with a single marker (QAMS) method with baicalin as the internal reference substance was established. The 2, 2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS⁺) method was applied to determine the antioxidant activity of different batches of Scutellaria baicalensis Georgi. Grey relevance analysis was conducted to calculate the grey correlation coefficients of each antioxidant in the herb. The holistic quality of 75 batches of collected samples from different product regions was evaluated to distinguish different commercial types of Scutellaria baicalensis Georgi. The relative correction factor (RCF) of each index component with baicalin as the internal reference indicated good applicability, and there was no significant difference in assay results between the QAMS method and the external standard method (ESM). The results of grey correlation analysis showed that the content of eight index components including baicalin correlated strongly with the antioxidant activity of Scutellaria baicalensis Georgi. The calculated grey relational coefficient ranged from 0.772 4 to 0.808 6. The content of oroxylin A-7-O-β-D-glucuronide, norwogonin-7-O-β-D-glucuronide, baicalein, wogonin and oroxylin A showed positive correlations with the antioxidant potency. In contrast, baicalin, wogonoside and viscidulin Ⅲ content exhibited negative correlations. The ratio of the flavonoid glycosides and flavonoid glycoside aglycones were found to be a critical factor to distinguish the different types of commercial herbs. The crude herbs could be characterized as Scutellaria baicalensis pith-decayed with a G/A ratio less than 10, and the ratio in Scutellaria baicalensis pith-not decayed should be more than 10. The QAMS method established in this study is simple and accurate, and can be used for the determination of eight flavonoids in Scutellaria baicalensis Georgi. Combined with ABTS⁺ antioxidant activity and grey correlation analysis, a comprehensive model for Scutellaria baicalensis Georgi quality determination based on biological activity was established, which provides new ideas and methods for the establishment of quality standards consistent with the characteristics of traditional Chinese medicine.

Twenty compounds were isolated and purified from the fresh roots of Huaizhong NO.1 Rehmannia glutinosa by various chromatographic techniques such as Toyopreal HW-40C, Sephadex LH-20, silica gel and semi-preparative HPLC. Their structures were identified by analysis of physicochemical properties and spectral data, and determined as 3-methyl-2-(hydroxymethyl)-4H-pyrone-4-one (1), 3-amino-2-pyridine ethanol (2), (5'S)-2-oxo-N-phenylpyrrolidine-3-carboxamide (3), L-pyroglutamic acid methyl ester (4), indazole (5), uridine (6), adenosine (7), rehmanalkaloid C (8), 1-(3-ethylphenyl)-1, 2-ethanediol (9), (3-ethylphenyl)-1, 2-ethanediol (10), 2-hydroxymethylphenyl-1-O-β-D-glucopyranoside (11), mussaenoside (12), 11-methylforsythide (13), 7-deoxygardoside (14), 1-O-α-L-rhamnopyranosyl(1→6)-β-D-glucopyranoside (15), 6-deoxy-D-mannono-1, 4-lactone (16), monoethylfumarate (17), daphneresinol (18), dihydroquercetin (19), rhamnopyranosyl vaniloyl (20). Compound 1 is a new compound, named as 3-methyl-2-(hydroxymethyl)-4H-pyrone-4-one, compounds 2 and 3 are new natural products, and compounds 4, 5, 9-11, 13, 14 and 16-19 are isolated from Rehmannia for the first time.

In recent years, many biosimilars have been approved marketing authorization in our country. When conducting research and evaluation of biosimilar, the quality similarity between the candidate drugs and the original drugs is the key point of "biosimilar", meanwhile it is the basis for clinical and marketing authorization of biosimilar. However, the industry and regulatory agencies are facing many challenges in establishing quality "similarity assessment criteria", such as: limited batches of original drugs; quality drift caused by manufacture process changes in the life cycle; analytical method of critical quality attributes is different from different manfactures; statistical methods used for establishing assessment criteria are difficult to unify. In the article, the data obtained from 69 lots of chemistry, manufacturing and control (CMC) dossier of trastuzumab originator company and dossiers of 9 biosimilar companies were analyzed. Furthermore, combined with the risk identification of the critical quality attributes of the product, quality "similarity assessment criteria" have been proposed. This standard has been verified by the disclosure of biosimilar quality data, aim to promote the development and evaluation of trastuzumab biosimilar.

Polymeric micelles have become a potential drug carrier. However, some single-copolymer micelles often have some performance defects, such as low drug loading, poor stability and weak response to microenvironment. Mixed micelles self-assembled from two or more dissimilar macromolecular polymers provides a direct and convenient approach to improve drug delivery. This paper attempts to review the internal force, preparation and characterization, performance advantages and biological activity of mixed polymer micelles, aims to show the advantages of polymer mixed micelles, analyzes the problems and shortcomings, and hopes to provide reference for the development of anti-tumor nano-micelles.

We aimed to explore the involvement of Fas-associated death domain protein (FADD) in the inhibitory effects of etoposide (VP16) on the proliferation, migration, and apoptosis of A549 non-small cell lung cancer (NSCLC) cells. FADD knockout (KO) and control A549 cells were constructed using the CRISPR/Cas9 system. The cell counting kit-8 (CCK-8) assay, the scratch wounding assay, and the Annexin V/PI staining-based flow cytometry were used to assess the effect of FADD KO on viability, migration, and apoptosis of A549 cells with or without the presence of etoposide, respectively. The expression pattern of several proteins involved in proliferation[raf proto-oncogene serine/threonine-protein kinase (c-Raf) and extracellular signal-regulated kinase of phosphorylation (p-ERK)], apoptosis[B-cell lymphoma 2 (BCL2), cleaved cysteinyl aspartate specific proteinase 3 (cleaved-caspase-3), and cleaved-caspase-9] and migration[matrix metalloproteinase 2 (MMP2)] was detected by Western blot. We found that FADD KO attenuated proliferation and migration of A549 cells. Consistently, we demonstrated that FADD KO enhanced etoposide-mediated inhibition of proliferation and migration in A549 cells. We further demonstrated that FADD KO obviously enhanced etoposide-mediated apoptosis in A549 cells. For mechanism exploration, we found that etoposide sensitivity enhanced by FADD KO may be partly explained by reduced expression of c-Raf, p-ERK, MMP2, and increased cleavage of caspase-3 and -9. Combined with the Kaplan-Meier (KM) survival curve analysis obtained from the GEPIA database, it is preliminarily judged that patients with high FADD gene levels in lung adenocarcinoma have a poor prognosis. Our study suggests that FADD can be used as a potential biomarker for the treatment of lung adenocarcinoma, providing a personalized treatment plan for the treatment of lung adenocarcinoma.

Twenty-five candidate MLO genes of Lonicera japonica were obtained by whole genome sequencing. Bioinformatics analysis showed that the number of amino acids in the protein family ranged from 137 to 846, the theoretical isoelectric point ranged from 5.02 to 9.50, and it was rich in basic amino acids. One protein did not contain transmembrane domain, and the other proteins ranged from 3 to 10. The results of subcellular localization showed that 21 proteins were located on the cell membrane and 1 protein was located on the chloroplast. Phylogenetic trees were constructed from 133 MLO proteins of Lonicera japonica, Triticum aestivum, Arabidopsis thaliana, Solanum lycopersicum, Nicotiana tabacum, Nicotiana sylvestris and Nicotiana tomentosiformis. The results indicated that MLO family proteins of Lonicera japonica could be divided into five subgroups. Tissue specific analysis showed that the expression of MLO genes in Lonicera japonica had obvious tissue specificity. Among them, 8 genes were highly expressed in leaves, 2 genes in stems and 2 genes in flowers. Four genes were significantly upregulated after inoculation with powdery mildew, among which MLO14 increased the most, which was more than 2 000 times higher than that of the control. In this study, the composition and expression of the MLO gene related to the occurrence of powdery mildew of Lonicera japonica were preliminarily analyzed, which laid a foundation for the further use of MLO as a target gene to develop new germplasm resistant to powdery mildew of Lonicera japonica.

Compared with systemic administration such as oral delivery or injection, inhaled medicines directly locate in the respiratory tract to exert therapeutic effects, offering obvious advantages in the treatment of respiratory diseases. Marketed inhaled medicines are yet difficult to meet the clinical demands, and there are considerable challenges in the discovery and development of novel inhaled medicines due to the lack of experiences- and property-based rules for inhaled compounds. Personalized modification of candidate drugs through prodrug technology to meet the requirements of inhalation therapy is the current alternative approach for inhaled drug development. In this review, we intend to summary the applications of prodrug technology in the research of inhaled medicines over the past 20 years. These studies have shown that esterified prodrugs and macromolecule conjugates could effectively prolong lung retention; mannose modification or acid-sensitive bond connection can achieve targeted drug release in alveolar macrophages; personalized modified prodrugs can obtain suitable physicochemical properties for pulmonary delivery and reduce drug toxicity. In general, the application of prodrug technology can modify the physicochemical and biopharmaceutical properties of drugs and may promote the discovery and development of novel inhaled medicines.

High altitude pulmonary edema (HAPE) is a result of leaking of the fluids from blood vessel to the lung tissue due to pulmonary artery hypertension in the high altitude place, which happens very quickly and shows high mortality. Sildenafil (SIL) can prevent HAPE by expanding pulmonary vessels; however, only oral tablets and injections of SIL are currently available. The formulations have the disadvantages of high doses, inconvenient use, and high systemic side effects. Here, liposomal sildenafil (LS) was prepared for pulmonary delivery and the prevention of HAPE was explored. The ammonium sulfate gradient method was used for the preparation of LS with a high entrapment efficiency of nearly 100%, the particle size of 116.97 nm, and the zeta potential of -30.93 mV. All animal experiments had been approved by the Ethics Committee of Academy of Military Medicine, Academy of Military Sciences, and carried out in accordance with relevant guidelines and regulations. The dose (3 mg·kg^-1 SIL) of sildenafil suspensions, LS or blank liposomes was intragastrically or intratracheally administered to the lungs of mice. The mice with or without treatment were put in the hypobaric oxygen chamber of 5 000 m altitude for 1 h. In the open field text the model mice had the shorter total distances and longer dead time than the healthy mice and they showed the lower percutaneous oxygen saturation (SpO₂), the higher tumor necrosis factor-α (TNF-α) levels of and the lower reduced glutathione (GSH) in the lung tissues. By contrast, oral LS remarkably modified the moving ability of the mice and they had the higher SpO₂ than the model mice and the similar TNF-α and GSH levels to the healthy mice. Sildenafil suspensions and LS with the same dose (3 mg·kg^-1 SIL) were intratracheally administered to mouse lungs, respectively, and the mice of the model group and the treatment groups were put in a hypobaric oxygen chamber for predetermined time. In the rotating rod experiments, the mice in the model group showed shorter drop latency and more drop times than those in the healthy group, indicating that the physical activity of the mice was improved due to treatment. The inflammatory cytokines, TNF-α and interleukin-1β (IL-1β) in the blood of model mice were higher than those from the healthy group and they decreased after treatment, where the LS group maintained the lowest level close to the normal level. IL-1β in the blood of mice in the LS group was lower than that in the SIL group (P < 0.05) 1 hour and 48 hours post-hypoxia. Hypoxia inducible factor-1α (HIF-1α) in the lung tissues of model mice increased but decreased to the normal range after treatment. Moreover, HIF-1α in the LS group was lower than that in the SIL group, where the values were significantly different 48 hours post-hypoxia (P < 0.01). Inhaled liposomal sildenafil is a promising medication for the prevention of HAPE.