Three-dimensional (3D) printing technology is a rapid prototyping technology for designing 3D models with special shape and complex internal structure via computer-aided/controlled drawing and preparing. This technology displays the characteristics of flexible processing, rapid shaping, low operating cost and high reliability. 3D printing technology may provide new strategies and approaches for the generation of a variety of new drug delivery systems, which makes its application in pharmaceutics attractive. This review briefly introduces the process and feature of 3D printing technology in preparation field and mainly introduces the research progress in the design and engineering of related preparations in the aspects of rate-controlled drug release, time-controlled drug release and targeted drug release. The prospects and challenges of 3D printing technology in the formulation engineering are analyzed.

Supersaturated drug delivery systems (SDDS) are defined as systems that are able to generate and maintain a sustained drug supersaturation in the gastrointestinal tract, facilitating the oral absorption of drugs with poor water solubility. Supersaturated drug solution is generated from a higher energy form of the drug or rapid dissolution through various formulation options. However, supersaturated solution is a thermodynamically unstable system that can easily lead to drug precipitation, missing the aim of improving the absorption. Therefore, maintenance of the supersaturated state is essential for the development of SDDS. Polymer-based SDDS take polymers as the precipitation inhibitor, which can effectively prevent the precipitation of drugs, generating an excellent effect on maintenance of the stability of supersaturated solution. However, different polymers have distinct anti-precipitation ability, and the mechanisms of such activity supported by the polymer remain unrevealed. In this review, we summarize the research advances in the absorption-enhancing mechanisms and in vitro evaluations of polymers-based SDDS. This review provides a reference for the design of rational SDDS.

As an important drug carrier, liposome has the advantages of high biocompatibility and low immunogenicity. It has been widely used in the field of drug delivery, especially the targeted treatment of tumors. However, traditional liposomes are composed of flowing dynamic phospholipid membranes, which are easy to fuse together, resulting in aggregation and drug leakage. In addition, the lower degree of polyethylene glycol (PEG) modification also limits the targeted delivery performance of the vector in vivo. In view of the problems, a nanoparticle-targeted drug delivery system combining the inorganic carrier calcium phosphate with liposomes was designed, namely lipid calcium phosphate (LCP). Using doxorubicin (DOX) as a model drug, doxorubicin-loaded lipid calcium phosphate nanoparticles (DOX/LCP) were prepared by reverse microemulsion method, and the preparation conditions were investigated. The structure and morphology of calcium phosphate cores were observed by infrared spectroscopy, EDS spectroscopy, and transmission electron microscopy. The particle size, encapsulation efficiency, drug loading, stability and release behavior in vitro of DOX/LCP were investigated. Confocal microscopy and flow cytometry were used to qualitatively and quantitatively evaluate the uptake of DOX in drug-resistant tumor cell line MCF-7/DOX by LCP, respectively, and the thiazolium MTT colorimetric method was used to examine its cytotoxicity. LCP exhibited a typical core-shell structure with good size uniformity and dispersibility. The particle size was in (48.6 ±3.9) nm, the potential was in (-12.1 ±1.2) mV, and the encapsulation efficiency was above 80%. Moreover, it has a good stability in simulated plasma. In vitro release of LCP had a significant pH dependence. When the pH of the environment was 7.4, the cumulative release within 24 hours was less than 20%; as the pH of the release medium decreases, the release rate of DOX/LCP was accelerated gradually. Accumulated release over 24 hours exceeded 90% in the pH 4.5 medium. LCP significantly promoted the uptake and accumulation of DOX by drug-resistant cells, and the inhibition rate of drug-resistant tumors was significantly increased in vitro. The half maximal inhibitory concentrations (IC₅₀) of LCP/DOX and free DOX were 4.6 and 11.8 μg·mL^-1, respectively, and there was a significant difference between the two groups (P < 0.05). In summary, the LCP prepared in this study had a small particle size, high encapsulation efficiency and good stability. It had environmental responsiveness and potential inhibition of tumor drug resistance, which suggests a potential in the clinical application.

The study was aimed to identify the related substances of vortioxetine hydrobromide by hyphenated techniques. The separation of the six related substances was performed on a Phenomenex Luna Phenyl- Hexyl column (150 mm×4.6 mm, 3 μm) by linear gradient elution of acetonitrile and ammonium formate solution. Electrospray and atmospheric pressure chemical ionization were interfaced respectively with high resolution Q-TOF/MS for the determination of the accurate mass and elemental composition of the parent ions of the related substances, and triple quadrupole tandem mass was employed for the product mass spectra determination. The structures of the related substances were identified through elucidation of the fragment ions. Vortioxetine hydrobromide and its related substances were adequately separated under the established HPLC conditions. Six major related substances were detected and identified for the first time. The data provides a reference for optimization of the synthetic process and quality assurance of vortioxetine hydrobromide.

In this study, the effects of honokiol (HN) treatment for 24 h on lipid synthesis was examined in HepG2 cells. The parameters include intracellular lipid droplet and the expression of SREBP-1c and PNPLA3, glucose uptake, and oxidative stress including the expression of CYP2E1 and CYP4A in normal, TO901317 (TO)- and oleic acid (OA)-treated HepG2 cells. The lipid droplets were detected by oil red O staining. The glucose uptake was measured by fluorescence spectrophotometry using[2-(N-(7-nitrobenz-2-oxa-1, 3-diazol-4-yl) amino)-2-deoxyglucose, 2-NBDG] as probe. The expression levels of target genes were detected by quantitative PCR and Western blot. The results showed that:① TO (5 μmol·L^-1) and OA (0.5 mmol·L^-1) treatment increased the levels of intracellular lipid accumulation and the mRNA and protein expression of SREBP-1c and PNPLA3. After HN (10, 20, 40 μmol·L^-1) treatment for 24 h, the lipid accumulation and the expression of SREBP-1c and PNPLA3 were all decreased in the tested cells. ② OA treatment significantly suppressed glucose uptake, while HN treatment dose-dependently increased the glucose uptake in OA-treated cells. ③ Compared with control group, CYP2E1 protein level significantly decreased in the three tested cells, and CYP4A protein level significantly decreased only in OA-treated cells following HN treatment. The above results suggest that HN may attenuate lipid accumulation by suppressing the expression of SREBP-1c and PNPLA3, and reduce lipid peroxidation and insulin resistance by down-regulation of the protein levels of CYP2E1 and CYP4A in HepG2 cells with steatosis.

MCT is an important key enzyme in the terpenoid biosynthesis in MEP pathway. In this study, Gateway technology was used to construct RNAi vector of TwMCT, and a vector fragment with a size of 484 bp was obtained. The TwMCT RNAi vector was transferred into the suspension cells of Tripterygium wilfordii by gene gun. Accumulation of terpenoids was assayed by UPLC, and the result showed that the content of triptolide and celastrol in cells decreased by 23.4% and 42.8%, respectively, compared with the control group pK7GWIWG2D. Moreover, the gene expression of TwMCT and major genes in terpenoid biosynthesis pathway was detected by qRT-PCR, which demonstrated that the expression of TwMCT reduced by 29.2% relative to that of the control group pK7GWIWG2D, and the relative expression of TwDXR, TwGGPS, TwHMGR and TwHMGS diminished by 36.3%, 31.3%, 62.2%, and 29.1%, respectively, but the expression of TwDXS was up-regulated by 114.2%, and there was no significant change in TwFPS. Thus, it was verified in vivo that interference with TwMCT expression significantly inhibited the accumulation of triptolide and celastrol in Tripterygium wilfordii, laying a foundation for further exploring the regulation mechanism of MCT gene on the terpenoid biosynthesis in Tripterygium wilfordii.

The commonly used traditional Chinese medicines Curcumae Rhizome (Ezhu) and Curcumae Radix (Yujin), are representatives of multi-plant sources. The relationship among the original source plant species is intricate. In this study, by using multivariate data analysis, volatile metabolites in rhizomes and radixes of source plants of Curcumae Longae rhizome (Jianghuang), Yujin and Ezhu in traditional Chinese medicine were compared and analyzed. The source plants included Curcuma longa, Curcuma kwangsuensis, Curcuma wenyujing and Curcuma phaeocaulis. The results indicated that:① volatile metabolites were similar in quality but variation in quantity for rhizomes and radixes origin from the same plant species; ② volatile metabolites of C. longa rhizomes showed bigger difference compared with others; ③ although common volatile metabolites were observed in rhizome of C. kwangsuensis, C. wenyujing and C. phaeocaulis, the difference among them were significant; and ④ significant differences were observed for the four kinds of radixes. Results in this study revealed the differences of the four source plants species, and similar metabolites in source plants of Curcumae Rhizome (Ezhu) and Curcumae Radix (Yujin) from the level of volatile metabolites. These results provided a reference for the clinical use of the three kinds of traditional Chinese medicine.

Dammarenediol-Ⅱ is an important precursor in the biosynthesis pathway of ginsenosides which are the main active components of Panax quinquefolius and Panax ginseng. For constructing a dammarenediol- Ⅱ-producing cell factory, the triterpenoid precursors of yeast are improved significantly by the modular pathway engineering strategy on the basis of an MVA optimized strain. The strain overexpressing Salvia miltiorrhiza SmFPS and Arabidopsis thaliana AtSQS2 could yield 67.4 mg·g^-1 squalene, accounting for about 6.74% of cell dry weight. In our further work, an Arabidopsis thaliana 2, 3-oxidosqualene synthase AtSQE2 was found to be able to increase the downstream lanosterol yield by 22-fold, reaching 47.9 mg·g^-1. Then, regulating dammarenediol-Ⅱ synthase gene expression, using anti-sense RNA technology for regulation of ERG7 in the ergosterol pathway, and optimizing fermentation process were successively performed. Finally, the synthesis flux of triterpenes was increased to 10 g·L^-1 for the first time, and we constructed an efficient cell factory that can produce 15 g·L^-1 dammarenediol-Ⅱ, which lays a solid foundation of industrial synthesis of dammarane-type ginsenosides.

Tripterygium wilfordii 3-hydroxy-3-methylglutaryl coenzyme-A reductase (TwHMGR) is an important regulation site in terpenoids metabolic pathway in cytoplasm which is the first speed limit enzyme of MVA pathway. In order to investigate the effects of TwHMGR on the biosynthesis of triptolide and celastrol in Tripterygium wilfordii, the overexpression of TwHMGR (OE-HMGR) was studied in this paper. We cloned the full-length of TwHMGR to construct overexpression vector by Gateway technology then delivered the expression vector into Tripterygium wilfordii suspension cells by gene gun. qRT-PCR was used to detect the expression of TwHMGR:the expression of TwHMGR was increased to 1.75 folds over the control group (empty vector:pH7WG2D) in the overexpression group. The accumulation of triptolide and celastrol in the suspension cells of Tripterygium wilfordii was detected by UPLC, revealing that:the contents of triptolide and celastrol were increased to 163.93% and 190.04% of over the control group in the overexpression group. Based on these findings, the positive effect on the accumulation of active terpenoids, triptolide and celastrol in Tripterygium wilfordii was found and the results laid a foundation of the synthetic biology research on important active terpenoids in Tripterygium wilfordii.

Plant flowering regulation is an important mechanism to response to environmental stress. Heat shock protein 70 family is one of the main molecular chaperones to resist stress; miRNA can be used as a negative regulator to participate in post-transcriptional gene in flowering network. In this paper, we obtained an Hsp70 gene from Lonicera japonica transcriptome and combined with Lonicera japonica miRNA library to obtain a novel miRNA that may target Hsp70 gene through bioinformatics method. Bioinformatics and expression during different flowering stages of the obtained Hsp70 gene and miRNA were analyzed. Phylogenetic tree showed that the obtained Hsp70 gene was clustered with Hsp110 subfamily in Oryza sativa and Arabidopis thaliana. The prediction of miRNA secondary structure showed its stable structure and high reliability. The binding site map showed that there were two base mismatches between sequences of miRNA and Hsp70 gene. The expression analysis showed that the expression of Hsp70 and miRNA in different flowering stages had opposite trends, indicating that miRNA might regulate Hsp70 to participate in the flowering stages of Lonicera japonica. This study provided new ideas for Lonicera japonica flowering regulation and response to environmental stress mechanisms.