Latest ArticlesA simple and efficient method for extracting Cistanche polysaccharides was obtained and used to extract four Cistanche polysaccharides. The molecular weight, distribution and polysaccharide mass fraction of the four Cistanche polysaccharides were measured by High performance size exclusion chromatography-multiangle laser light scatter-refractive index detector (HPSEC-MALLS-RID) and differences between the four Cistanche polysaccharides were determined. The results of a single factor test and an orthogonal test showed that the best process parameters for extracting Cistanche polysaccharides used a mixture of enzyme (4%), a ratio of complex enzyme (cellulase∶pectinase) 1∶1, ultrasonic power at 350 W (40 kHz), for 20 min, enzymolysis temperature of 50 ℃, and pH 6. The average yield of three repeated extractions was 5.14%, the mass fraction was 90.78%, the comprehensive index was 47.96%, and the RSD = 1.51%, indicating good reproducibility. The four Bossica Cistanche polysaccharides are all spherical structures but their molecular weights and distributions differ. The optimized compounded enzyme combined with ultrasonic extraction increases the yield of Cistanche polysaccharides. It is a simple, convenient, economical and environmentally friendly extraction method, which lays the foundation for future polysaccharide research.
The study was undertaken to clarify the differences in metabolite groups between Aster yunnanensis and Pulicaria insignis to establish plant origin and identify plant resources. Non-target metabolomics of A. yunnanensis and P. insignis was undertaken with sample derivatives and gas chromatography-mass spectrometry (GC-MS). Principal component analysis (PCA), partial least-squares discriminant analysis (PLS-DA) and orthogonal partial least squares-discriminant analysis (OPLS-DA) were used to characterize the differing metabolites of A. yunnanensis and P. insignis. Correlation analysis of these metabolites and metabolic pathway analysis allowed us to identify metabolic pathway changes between samples. Characterization of the antibacterial activity of the essential oil from A. yunnanensis and P. insignis showed that 384 metabolites were identified in the two species, including amino acids and peptides, phenylpropanoids, aromatics, glycoside, nucleotide, flavonoid, alkaloids, saccharides, vitamin, organic acid, lipids, esters, alcohol. A total of 92 differential metabolites with significant differences in content (P < 0.05, VIP > 1) were identified. In conclusion, a new method based on pre-column derivatives and GC-MS metabolomics was used to distinguish and compare A. yunnanensis and P. insignis metabolites.
This article intended to build a component-target network to screen the effective components of Compound Suanzaoren Decoction, and to establish an efficient and rapid method for the determination of effective components. 103 compounds in the Compound Suanzaoren Decoction were identified by HPLC-Q-TOF MS/MS. 27 potential effective substances were analyzed by network pharmacology. The first six components with the largest degree were selected as effective components, including neomangiferin, mangiferin, ferulic acid, senkyunolide Ⅰ, spinosin, and 6'''-feruloylspinosin. A UHPLC method was established for determination. The chromatographic separation was performed on a Waters CORTECS T3 column (2.1 mm×150 mm, 1.6 μm) with the mobile phase of 0.1% phosphoric acid solution and acetonitrile. The results showed that the specificity was good with no interference for the chromatographic peaks. The linear was excellent within the scope of investigation, with the values of r higher than 0.999 0 for all analytes. The method had been verified to have good sensitivity, precision, accuracy, durability and stability. Therefore, the results showed that the method established was suitable for the screening and determination of effective components in Compound Suanzaoren Decoction, which provided a basis for the quality evaluation of traditional Chinese medicine.
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.
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.
Hesperidin nanosuspensions (HDN-NS) were prepared with tea saponin (TS) as stabilizer. The feasibility of TS as a natural stabilizer and the mechanism of TS stabilized nanosuspensions were investigated to provide reference for the development of green nano-preparations of hesperidin. HDN-NS was prepared by high-speed shearing combined with high-pressure homogenization. Using average particle size and polydispersity index as evaluation indexes, the effects of drug concentration, shearing speed, shearing time, homogeneous pressure and homogeneous cycles on HDN-NS were investigated by single factor experiment. The results of single factor investigation were as follows: drug concentration was 8.0 mg·mL-1, shearing speed was 16 000 r·min-1, shearing time was 2 min, the homogeneous cycles were 6 cycles at 35 MPa and 12 cycles at 100 MPa. The pH, ionic strength, zeta potential and critical micelle concentration of TS were investigated to determine the role of electrostatic repulsion in the mechanism of TS stabilized nanosuspensions. The results showed that electrostatic repulsion is involved in the mechanism of TS stabilized HDN-NS. In conclusion, at low concentration, TS can significantly reduce the particle size of HDN-NS, which indicates that TS has the potential to stabilize nanosuspension. Electrostatic repulsion is one of the mechanisms of TS stabilizing nanosuspension.
Clarithromycin (CLA) belongs to the second generation macrolide antibiotic. The commercial crystal form of CLA is form Ⅱ, and it exhibits the poor compressibility during the tablet pressing process. Since the crystal form of drugs has a significant effect on their mechanical properties, from the perspective of crystallography, this study investigated the difference of the compressibility between CLA form Ⅰ and form Ⅱ, and analyzed the mechanism that led to such difference. On the other hand, from the perspective of pharmaceutics, we also studied the effect of filler on the compressibility of form Ⅱ. It was found that CLA form Ⅰ had improved plastic deformation than form Ⅱ because of the slip planes in its crystal structure leading to the better compressibility. Moreover, microcrystalline cellulose, pre-gelatinized starch and lactose monohydrate could improve the compressibility of form Ⅱ, and microcrystalline cellulose showed the best effect. We improve the compressibility of CLA from crystal form and filler, and also lay a foundation for the development of CLA solid preparations.
The MIKC-type MADS-box gene family plays an important role in flower development in plants. This study identifies members of the MIKC-type gene family in Cannabis sativa L. at the genome level, with the chromosomal location and linkage, evolutionary relationships, and identification of conserved motifs determined using bioinformatics tools. The results show that C. sativa contains 39 members of the MIKC-type MADS-box gene family (named CsMADS1-CsMADS39) unevenly distributed on nine chromosomes. The encoded proteins range in length from 146 to 503 amino acids, and the theoretical isoelectric points range from 5.19 to 10.12. Molecular weights range from 16 739.35 to 57 070.56 Da. The subcellular location of CsMADS genes is mainly in the nucleus. The result of conserved domain analysis showed that all genes contain the MADS conserved domain. The analysis of GO showed that all genes were annotated to 331 GO terms, which were clustered by molecular function. A phylogenetic tree showed that CsMADS genes could be divided into 14 subclasses, according to ABCDE homologous gene model; CsMADS genes are clustered with SQUA/AP1, AP3, PI, GGM13, AG, SEP/AGL2 subfamilies in Arabidopsis. There are abundant cis-acting elements in the upstream promoter region of CsMADS genes, mainly light regulatory elements. The results of real-time fluorescent quantitative PCR showed that some CsMADS genes are highly expressed in flowers and bracts, with tissue-specific expression. This study identified and analyzed the MIKC-type MADS-box gene family in C. sativa at the genome level, and provides a theoretical basis for further exploration of the function of MIKC-type genes and their role in regulating the growth and development of medicinally important hemp.
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.
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.
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