The key of gene therapy is to deliver the functional gene to the target tissue in the body. The safe and efficient gene carrier is particularly important in the targeted delivery. Multifunctional envelope-type nano device (MEND), based on concept "Programmed packaging", is a new type of gene carrier system, with high encapsulation efficiency, favourable stability, high transfection efficiency, easy preparation, etc. MEND is designed to control intracellular trafficking as well as the tissue distribution of encapsulated compounds such as nucleic acids/proteins/peptides, permitting them to function at the appropriate location. In this paper, research progresses in MEND are reviewed in accordance with three types of payloads:the small interfering RNA (siRNA), DNA and proteins/peptides in recent years.

The study was designed to test the estrogen-like effects about allantoin. The activity of the allantoin was investigated by mouse uterine weight gain test and MCF-7 cell proliferation assay. The levels of E2, FSH and LH were also measured. ICI182, 780, MPP, THC and G15 antagonnist assay and Western blot were adopted to explore the mechanism of allantoin. Allantoin increased the uterus index of premature female mice, the levels of E2 and FSH, and the expression of ERα and GPR30, compared with the control group. Allantoin also promoted the proliferation of MCF-7 cells. Co-incubation of MCF-7 cells with estrogen receptor blockers, ICI182, 780, MPP and G15 abolished the inductive effect of the proliferation. These results suggest that allantoin has estrogenic activities, which are mainly mediated by ERα, GPR30.

The Chinese herbal Sophora alopecuroides is widely used to clean intestine and eliminate dampness, and it has good therapeutic effects on treating bacillary dysentery and inflammatory bowel disease, etc. in clinics. However, the mechanism of treatment is not yet well understood. The present study was aimed to explore the mechanism of Sophora alopecuroides treatment of large intestine dampness-heat syndrome (LIDHS). The LIDHS model was performed by the comprehensive factors, including high temperature and humidity environment, high-sugar and high-fat diet, and intraperitoneal injection of Escherichia coli. The blood routine, serum proinflammatory cytokine levels and histopathological changes of intestine were detected and observed. Meanwhile, the serum metabolomic approach was conducted using the method of ultra performance liquid chromatography coupled to quadrupole time-of-flight mass/mass spectrometry (UHPLC-Q/TOF-MS/MS). The results showed that Sophora alopecuroides has good therapeutic effects on the LIDHS rat models. After treatment with Sophora alopecuroides, the abnormality of blood routine indexes as well as proinflammatory cytokines, including IL-1β, IL-2, IL-6 and TNF-α in vivo, tended to be normal, and the histopathological changes of intestine were improved. Through metabolic profiling and protocol analysis, 9 potential metabolic markers may be closely related with the treatment mechanism of Sophora alopecuroides on this disease, including taurine, L-tryptophan, LysoPE, LysoPC, LPA, DG, chenodeoxycholic acid disulfate, traumatic acid and 7-ketodeoxycholic acid, which were involved in taurine and hypotaurine metabolism, glycerophospholipid metabolism, glycerolipid metabolism, tryptophan metabolism and primary bile acid biosynthesis etc. The serum metabolomic approach can be applied to clarify the therapeutic mechanism of Sophora alopecuroides on LIDHS, and provide the theoretical basis for Sophora alopecuroides in clinical practice.

Metabolic transformation in vivo is a critical approach in the study of toxicity, but real-time dynamic observation of the transformation is difficult. We proposed that zebrafish toxicity/metabolism synchronization may be used in the analysis of toxicity of Folium Epimedium (Yinyanghuo for Chinese, YYH) and the toxicity may be reduced by Radix Morindae Officinalis (Bajitian for Chinese, BJT). Healthy zebrafish embryos 1 day post fertilization (1 dpf) were exposed to different concentrations of YYH, total flavonoids of YYH (YTF), representative flavonoids (epimedin C and icariin) and their respective in combination with BJT. Death numbers of the embryos or larvals were counted during 1-5 days after dosing (2-6 dpf); embryonic micro-morphology of zebrafish (3 dpf) was observed and pictures were taken. The blank vehicle (0.4% DMSO) was used in the control group, and LC₅₀ value of 2 to 6 dpf was calculated by SPSS16.0. A relative safe concentration was sampled every day to analyze the dynamic metabolites changes of major flavonoids of YYH. The results showed that epimedin A/B/C (EA/EB/EC) and icariin, the major flavonoids of YYH, were dynamically transformed into major metabolites of sagittatoside C (SC) and baohuoside I (BI) by zebrafish. BI was mainly derived from EA, EB and icariin. Neither original form nor their metabolite BI can cause zebrafish poisoning. SC was mainly derived from EC, and its accumulation was closely related to the toxicity of YYH, YTF and EC. After combination with BJT, the metabolism of EC was slowed down and the toxicity was alleviated. Zebrafish toxicity/metabolism synchronization revealed that the toxicity of EC of YYH was increased after metabolism into SC, which maybe the key potential poisonous factor of YYH, and BJT can reduce the toxicity by slowing down the metabolism rate of EC. The data provides new ideas and methods in the study of toxic substances in Chinese medicine and mechanism of detoxicity by combination.

CRISPR/Cas9 system, consisting of clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated (Cas) proteins, is a prokaryotic immune system that confers resistance to foreign genetic elements such as those present within plasmids and phages. A simple version of the CRISPR/Cas system, type Ⅱ CRISPR, has been modified to edit genomes. By delivering the Cas9 nuclease together with a synthetic guide RNA (sgRNA) into cells, genome can be edited at desired loci site. CRISPR/Cas genome editing techniques have been widely implemented in various species and research areas. In this review, we summarize the several applications of CRISPR/Cas9 in the field of drug discovery and development, which include target gene screening and editing, drug target screening and validation, generation of animal models and treatment of genetic disease, etc. The defects and improvements of CRISPR/Cas9 technology is discussed as well.

This study was designed to explore the mechanism of Coix seed oil (Coix) impact on the drug resistance, bioluminescence imaging (BLI) and the efflux of D-luciferin potassium salt, the substrate of ABC transporters, in doxorubicin-resistant breast cancer cells. Multidrug resistance (MDR) gene and protein expression were analyzed in the cells by q-PCR and Western blot. First, in order to investigate the effect of the efflux function by ABC protein, a cell line with overexpressed luciferase was established in MCF-7 cell line. BLI was used to monitor the efflux kinetics of D-luciferin potassium salt before and after Coix treament. The results showed that the efflux of D-fluorescein potassium from MCF-7/DOX^Fluc was lessened when pretreated with Coix, which means that Coix may decrease the efflux of other chemotherapies using ABC transporters. Both of the results of q-PCR and Western blot showed that gene and protein expression of ABC transporters such as ABCG2, ABCC1 and ABCB1 were down-regulated by Coix treatment. The efficacy of Coix reversing MDR was verified with the chemotherapy medication doxorubicin (DOX). MTT assay showed that Coix increased the inhibitory effect of DOX on proliferation of MCF-7/DOX, and the optimal combination of ratio was 25 times that of DOX. The results suggest that Coix may reverse MDR of the substrate of ABC transporters from two aspects, one is to cut down the ABC protein efflux function, and the other is to decrease the quantity of ABC gene and protein expression.

Honokiol (HK) have extensive pharmacological activities, but its poor solubility and instability restricted its clinical application and efficacy exertion. HK nanosuspensions (HK-NSps) were designed in this study in order to solve the problems. HK-NSps were prepared by antisolvent precipitation method, using poly-vinylpyrrolidone (PVP) and bovine serum albumin (BSA) as a combined stabilizer. The particle size was measured using dynamic light scattering method, the morphology was observed by transmission electron microscopy. The size change and drug content of HK-NSps in various physiological media during the storage at ambient temperature was examined to evaluate their storage stability. Dialysis method was used to study their drug release in vitro. MTT assay was used to assess their in vitro cytotoxicity against 4T1 breast cancer cell line. Anti-tumor effect in vivo was also investigated in 4T1 tumor-bearing mice. HK-NSps were prepared with high drug loading content of 48.62%, nearly spherical shape and good storage stability. The average particle size was (83.40 ±1.042) nm, the polydispersity index (PDI) value was 0.223 ±0.011, the zeta potential was (-42.2 ±1.2) mV. HK-NSps showed sustained in vitro drug release and enhanced cytotoxicity in contrast to free HK against 4T1 cells (IC₅₀, 8.36 μg·mL^-1 vs 37.58 μg·mL^-1, P < 0.05). The in vivo study on 4T1 tumor-bearing mice demonstrated that HK-NSps showed good dose-dependent tumor inhibition rate (TIR). In contrast to 4 mg·kg^-1 of PTX injection (TIR, 47.9%), medium and high dose of HK-NSps displayed improved therapeutic efficacy (TIR, 55.67% for 40 mg·kg^-1, 67.28% for 60 mg·kg^-1, P < 0.05). In contrast, the high dose of HK crude drug (60 mg·kg^-1) had TIR of only 54.13% even administrated every day. In conclusion, HK-NSps were prepared with small size, high drug-loading capacity, and good stability. The improved in vitro and in vivo antitumor efficacy demonstrated that HK can be a promising antitumor drug in combination with nanosuspensions technology.

Currently, the specification grading standard for Astragali Radix can not accurately reflect growth years. The aim of this study is to identify the growth ring number of different parts of 1 to 6 year Hengshan imitative wild culture Astragali Radix, in order to get a different absolute growth years, to classify the accumulation rules of the content of flavonoids and saponins, and to lay the foundation for evaluating quality of Astragali Radix. Observing growth ring numbers of 1-6 years Astragali Radix by means of hand sections and paraffin sections in the study, and analyzing the number of different growth years and different diameter. At the same time, HPLC-UV-ELSD was used to analyze the 12 index components of the samples with absolute growth years of 2 to 6 years. The results indicated that the growth ring number excepting hollow part is consistent with the actual growth period of Astragali Radix and the number of growth rings gradually decreased from the upper to lower. The results of HPLC-UV-ELSD determination showed that the saponins content of 3-year-old Astragali Radix was the highest while the flavonoids content of the 4-year-old reached the maximum. The study provided the basis for foundation of the specification grading standard for Astragali Radix and clinical rational use drug.

Compared with the racemate of chiral drugs, enantiopure chiral drugs have been the hot spot of drug research because of their higher selectivity and lower side-effects. Although remarkable progress of asymmetric synthesis has been achieved in the last decades, chiral resolution is regarded as an important approach to obtain chiral drugs. Recent research advancements in the field of chiral resolution of racemic drugs and intermediates are reviewed here. It is clear that combination of chiral separation and racemization to improve the resolution efficiency has become a trend of chiral resolution. In addition, we also introduce some novel resolution methods, such as chiral extraction, membrane resolution, and resolution using nanoparticles.

A new dihydroflavone:mirabiflavone (1), together with two known compounds were isolated from the ethyl acetate extract of the roots of Mirabilis himalaica by using various chromatographic techniques, such as silica gel column, Sephadex LH-20 column, and semi-preparative HPLC. Their structures were elucidated as syringaresinol (2) and lariciresinol (3) by spectroscopic analysis. Compounds 2 and 3 were isolated from this plant for the first time.