This study aimed to evaluate the antioxidant activities of Scutellaria baicalensis Georgi flowers by measuring the scavenging activity of hydroxyl radical and superoxide anion radical. Aging of rats was induced by D-galactose and protective effects of Scutellaria flower extract (SFE) at different doses were evaluated by locomotor activities. To explore the mechanism of SFE, metabolomic technique combined with multivariate statistical methods was used to analyze the profile of metabolites from rat serum and quantify the levels of glutamine and glutamate in the serum. All research procedures were approved according to the Committee on the Ethics of Animal Experiments of Shanxi University. The results showed that high-dose of SFE had antioxidant activity in vitro, and could ameliorate the aging behavior of rats. A total of 9 potential biomarkers related to aging were found, pointing to the involvement of 7 metabolic pathways.

Carbon monoxide (CO) is an important chemical gas messenger molecule in the body with anti-inflammatory activity. As an active substance in gaseous state, the method for its safe and effective delivery towards the lesion sites remains to be established. Based on the natural affinity of carbon monoxide to hemoglobin, a main component of red blood cells (RBCs), this study proposes a carbon monoxide-red blood cell (CO-RBC) composite system, and tested its therapeutic effect against lung injury in an animal model. The mouse model of septic lung injury was adopted, and the carbon monoxide release molecule (CORM-2) was used as a positive control. CO-RBC system was characterized by CO release, stability, toxicity and in vivo lung targeting. The expression of intercellular adhesion molecule (ICAM-1) and pulmonary surfactant protein-A (SP-A) were evaluated in the animal model and the therapeutic effect of CO-RBC system for sepsis was measured by inflammatory factors tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6), as well as survival time of mice and pathological changes of the lung. Our results show that CO-RBC system exhibited satisfactory stability with negligible CO release during 48 h storage under nitrogen protection, while the CO release was about 70% within 12 h under physiological condition, in contrast to CO burst release from CORM-2. The CO-RBC system showed no significant toxicity in the animal model, and in vivo fluorescence imaging results showed effective accumulation in the lungs, supporting its lung targeting effects. The secretion of TNF-α and IL-6 in the CO-RBC group was significantly lower than that in other groups, the degree of pulmonary interstitial edema was relieved, the white blood cell infiltration was decreased, and the survival rate was significantly improved. Therefore, the CO-RBC system has a significant inhibitory effect on the pulmonary inflammatory response in septic mice compared with CORM-2. This system provides a new hope for therapeutic treatment of sepsis.

To investigate the influences of zwitterionic polymer chain length on mucus permeability and cellular uptake, the nanoparticles (NPs) were coated with poly(sulfobetaine methacrylate) (pSBMA) with different chain lengths. The di-block polymer poly(ε-caprolactone)-block-poly(sulfobetaine methacrylate) (PCL-pSBMA) with different chain lengths were synthesized via atom transfer radical polymerization (ATRP) combining with ring-opening polymerization of ε-caprolactone, and corresponding nanoparticles (pSBMAn NPs) were prepared by nanoprecipitation method. The sizes of different pSBMAn NPs were around 100 nm, and zeta potential were about -7 mV. Mucin interaction or mucus penetration study based on transwell systems were employed to evaluate mucus permeability of NPs. Caco-2 cells and mucus-producing HT-MTX-E12 cells were employed to illustrate the endocytosis efficiency of pSBMAn NPs. The results showed that the permeability coefficient of NPs coated with shorter chain length of pSBMA (pSBMA₁₀ NPs) was only 42.83% of that coated with longer pSBMA (pSBMA₈₀ NPs). On the contrary, the cellular uptake of pSBMA₁₀ NPs was 2.44 fold higher compared to pSBMA₈₀ NPs. Although the cellular uptake of pSBMAn NPs was reduced in the presence of mucus, pSBMA₁₀ NPs still presented the highest cellular uptake. However, the in vivo results indicated that the oral bioavailability of pSBMA₂₀ NPs was higher than that of pSBMA₁₀ NPs. All animal procedures were performed in accordance with the Guidelines of the Sichuan University Animal Care and Use Committee and were approved by the Animal Ethics Committee of Sichuan University. This study provides a reference for oral delivery of zwitterionic nanoparticles.

Melatonin (MLT) is an endogenous chemical that has antitumor effects at high doses. However, it shows low oral bioavailability and short in vivo half-life, leading to drug resistance. Here, liposomal melatonin dry powder inhalers (LMD) were prepared, and were used for treatment of primary rat lung cancer by pulmonary delivery. Liposomal melatonin (LM) was prepared by the ethanol injection method to achieve an entrapment efficiency of 98.89%. LMD was obtained by freeze-drying after LM was mixed with mannitol. LMD appeared as spherical particles under a scanning electron microscope. The rehydrated liposomes had a small size of 65.15 nm and the zeta potential of -14.2 mV without change inentrapment efficiency. LMD had an aerodynamic particle size of 6.73 ±0.012 μm and a fine particle fraction (FPF < 8.06 μm) of 22.2%, suitable for pulmonary delivery. When administered with the same dose, LMD showed much higher inhibition on A549 lung cancer cells than MLT and gemcitabine. LMD of a large dose had no effect on the growth of normal lung epithelial cells (BEAS-2B). Rat lung cancer models were established after 45 days by instilling 3-methylcholanthrene (MCA) and N, N-dimethylnitrosamine (DEN) into the rat lungs once (the experiments had been approved by the ethics committee and carried out in accordance with relevant guidelines and regulations). Decreases of tumor nodules and inflammatory cells in the tumor-bearing rat lungs were observed after treatment of MLT, gemcitabine and LMD by pulmonary delivery compared with the models, wherein LMD was most effective. The efficiencies of inhibition of NF-κB p65, increase of Tunel detection (indicating enhancement of apoptosis), and decrease of malondialdehyde corresponded to LMD being most effective. Therefore, given the fact that LMD can deliver the drug into the tumor tissues of lungs, and it presents as a promising pulmonary inhalable regiment for treatment of lung cancer.

Disulfiram (DSF) is a traditional anti-alcohol drug, but it was recently found that DSF has strong inhibitory effect on the growth of a variety of cancer cells. However, its clinical application is greatly limited due to its poor solubility, instability in gastrointestinal tract and short plasma half-life. In this study, DSF is fabricated into nanosuspensions with the aim of trying to solve these problems. DSF nanosuspensions (DSF-NSps) were prepared by the anti-solvent precipitation method under ultrasonication, and the suitable stabilizer was screened according to the size, polydispersity index (PDI), and zeta potential of the resultant nanosuspensions, along with their particle size change during the storage at room temperature. The particle size, PDI, and zeta potential of DSF-NSps were determined using dynamic light scattering method, while the morphology of DSF-NSps was observed by transmission electronic microscope (TEM). The stability of DSF-NSps in media was examined according to their particle size change in different physiological media. The concentration of DSF was measured by HPLC assay. The in vitro drug release was evaluated on basis of dialysis. MTT assay was employed to evaluate the in vitro cytotoxicity of DSF-NSps against cancer cell lines. The 4T1 tumor-bearing mouse model was used to evaluate the in vivo therapeutic efficacy of DSF-NSps. All the animal experiments were acquired according to the Regulations for Animal Experiments and Guidelines for Ethical as defined by Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences and Peking Union Medical College. As a result, the combinational use of soyabean lecithin (SPC) and D-alpha tocopherol acid polyethyene glycol succinate (TPGS) was determined to best stabilize DSF-NSps when the ratio of DSF-SPC-TPGS was 24:20:4 (weight ratio), with small particle size and good storage stability. The resultant DSF-NSps showed a regular spherical morphology and drug loading content of (45.36 ±2.09)%, with average particle size of 175.00 ±0.75 nm, PDI of 0.24 ±0.07 and zeta potential of -14.3 mV. DSF-NSps displayed good particle size stability in a variety of biological media including phosphate buffer saline, normal saline, 5% glucose, artificial gastric fluid, artificial intestinal fluid and plasma, which would meet the demand of both intravenous and oral administration. The in vitro study demonstrated that nano-encapsulation greatly increased the stability of DSF in aqueous media, DSF-NSps exhibited sustained release of the encapsulated drug and significantly inhibited 4T1 cells compared to free DSF (IC₅₀, 1.07 vs 5.53 μg·mL^-1, P < 0.01). DSF-NSps showed a good dose-response relationship on the 4T1 tumor-bearing mice with the tumor inhibition rates at the three doses being 80.22%, 75.14% and 66.10%, all higher than that of paclitaxel injections (55.01%, P < 0.05). The in vivo biodistribution study displayed that DSF-NSps were mainly distributed into liver, spleen and tumor. In sum, disulfiram nanoparticles could be expected to provide an effective anti-cancer drug for the treatment of breast cancer.

Tropinone reductase Ⅰ (TRI) is a key branch point enzyme in the midstream of tropane alkaloids (TAs) biosynthesis pathway and represents an important target for TAs metabolic engineering, which can lead to metabolic flux of substrate tropinone to TAs. A novel TRI gene was isolated from Datura arborea, a woody resource plant, and designated as DaTRI2 (GenBank accession number is MH705164). The full-length cDNA of DaTRI2 with 1 135 bp exhibits a high sequence homology (96.8%) with DaTRI, and is predicted to encode a protein of 347 amino acids. Deduced DaTRI2 protein contain a conserved TGXXXGXG motif involved in NADPH binding, the catalytic N-S-Y-K tetrad motif and eleven amino acid residues important for binding to its substrate tropinone. The phylogenetic analysis revealed that DaTRI2 and other TRIs from Solanaceous plants belong to the same cluster and DaTRI2 exhibited closest phylogenetic proximity to TRIs from Datura. DaTRI2 was expressed in E. coli and the purified recombinant protein can catalyze both tropinone reduction and tropine oxidation with an optimum pH value of 8.0 and 9.6, respectively. When tropinone was used as the substrate, the K_m and V_max values of DaTRI2 at pH 6.4 were 210.05 μmol·L^-1 and 69.6 nkat·mg^-1 protein respectively, while the K_m and V_max values for tropine as the substrate were 188.03 μmol·L^-1 and 114 nkat·mg^-1 protein respectively, at pH 9.6. DaTRI2 transcript was most abundant in the young leaf, followed by the root. Cloning of DaTRI2 gene and biochemical analysis of recombinant DaTRI2 facilitate further research on the molecular mechanism on TAs biosynthesis in woody plants and provide a more potent candidate for TAs metabolic engineering.

Q-marker is a new concept of quality control for traditional Chinese medicine. Since its introduction in 2016, it has received extensive support and participation from researchers and manufactures of the entire industry. In order to establishing the research level of quality markers, normative research model, we review the proposed core theory and research methods of Chinese medicine quality markers. The definition and scientific connotation of Chinese medicine quality markers are analyzed. The core theories and research methods of quality markers are summarized from five aspects:"effectiveness", "specificity", "transfer and traceability", "compatibility environment" and "measurability" of quality markers. This paper aims to provide useful theoretical and methodological guidance for studying Chinese medicine quality markers.

The quality of traditional Chinese medicine (TCM) is the lifeline for TCM industry. The development of artificial intelligence (AI) has provided new means for the quality management of Chinese medicinal materials (CMM). In this paper, we take the quality marker (Q-marker) as a breakthrough point, focused on the research strategy from chemical markers to Q-markers, picked up the characteristics of the Q-markers from the near infrared spectrum (NIRS), and explored the feasibility of establishing the NIRS assay based on Q-marker. After integrated the biological activity detection and artificial neural network algorithm, we try to establish the relationship between the spectral properties of NIRS and specific efficacy of the CMM. Finally, the bottlenecks will be solved that related to the transmission and traceability of quality attributes in the process of TCM production, quantity change, overall quality management and so on. This system is going to improve TCM quantity scientific and intelligent supervision, and promote the upgrading of traditional TCM industry.

The quality control of traditional Chinese medicine provides the premise of its modernization and globalization. Currently, the dual quality control based on chemical benchmark and effect benchmark has been recognized domestically and internationally. Research efforts have lead to establishment of a series of effective quality control methods based on chemical components, medicinal properties, microscopic characteristics, material constituents and pharmacodynamic targets. In the study of quality control based on chemical benchmarks, fruitful results on fingerprints, DNA barcodes, and quality markers have been achieved. However, due to a variety of factors, such as growth period, origin, growth environment and preparation process of traditional Chinese medicine, the quality control of traditional Chinese medicine based on chemical benchmarks remains difficult to fully reflect the quality of traditional Chinese medicine. At present, there is still a dispute on how to accurately reflect the quality of traditional Chinese medicines based on chemical benchmarks. For example, the index components selected in the Chinese medicine quality standards are difficult to totally reflect all the components of Chinese medicine, and the relevance between the index components versus therapeutic effect is not yet clear. In view of the complex signal network by cascade reaction and crosstalk of multi-signaling pathways within an organism, and the coordinated regulation of multi-components and multi-targets of traditional Chinese medicine, there may be different components regulating the same signal network or situations where the amount of certain chemical components within a range is not sufficient to cause a change in the signal network. Therefore, the quality control of traditional Chinese medicine based on the effect benchmark may be a useful supplement to the quality standard of traditional Chinese medicine. This paper proposes a Q-biomarker research strategy based on the effect benchmark in order to provide a methodological reference for the quality control research of traditional Chinese medicine.

The quality definition of traditional Chinese medicine (TCM) is a hot area in modern research of TCM. In recent years, the characteristics of one herb with multiple effects have been widely accepted and studied. The typical opposite-effect of herbs is considered as a special part of one herb with multiple effects, and was summarized in this paper. Sanqi was used as an example of opposite-effect herbs for developing the strategies and approaches on the Q-markers. The traditional opposite-effect should be studied by modern pharmacological research methods. The correlation of the chemical components with the opposite effects should be established in order to verify the material basis and evaluate the mechanism including targets and pathways. The unique characteristics of chemical components should be analyzed and defined. Finally, the Q-markers of the opposite effect herb will be confirmed. This paper provides a useful reference for the precise quality control of herbal opposite-effects.