The p21-activated kinase 1 (PAK1) is a member of the P21-activated protein kinase family that plays an important role in the proliferation and on cogenesis of pancreatic cancer. PAK1 is an important target for the treatment of pancreatic cancer. At present, akinase inhibitor targeting PAK1 is still in the preclinical research stage. Therefore, screening for new PAK1 kinase inhibitors is of great significance. In this study the natural compound celastrol was found to have a significant inhibitory effect on PAK1, with an IC₅₀ value of 3.614 μmol·L^-1. Molecular docking results showed that celastrol had good binding to PAK1. An MTT assay indicated that celastrol inhibited the proliferation of pancreatic cancer cells BxPC-3 and PANC-1. Mechanistic studies revealed that the inhibition of pancreatic cancer cells by celastrol was reversed by PAK1 siRNA. Celastrol inhibited PAK1 and the subsequent activation of downstream signaling pathways, thereby activating apoptosis signaling pathways and triggering apoptosis in pancreatic cancer cells. These findings suggested that celastrol induced apoptosis in pancreatic cancer cells by suppressing the PAK1 kinase signaling pathway and has potential value for the treatment of pancreatic cancer.

In this study a D-galactose-induced aging rat model combined with ¹H NMR of serum and liver metabolomics were used to explore the anti-aging effect and the potential metabolic regulatory mechanism of Scutellaria baicalensis Georgi leaves. All procedures involving animal treatment were approved according to the Committee on the Ethics of Animal Experiments of Shanxi University. The results of physical characteristics, an open field test and serum biochemical indexes indicated that Scutellaria baicalensis Georgi leaves had an anti-aging effect that could ameliorate the characteristics of aging rats such as acquired hair loss and slow response, improve the spontaneous activity of aging rats, and decrease lipid peroxidation and glycosylation damage induced by D-galactose. Serum and liver metabolomics further revealed that Scutellaria baicalensis Georgi leaves could decrease serum and liver metabolism disturbances in aging rats, mainly through different metabolites and metabolic pathways. Specifically, 12 differential metabolites including glutamine and glutamate, 11 metabolic pathways including D-glutamine and D-glutamate metabolism and alanine, aspartate and glutamate metabolism in serum were significantly altered after the treatment. Simultaneously, five differential metabolites such as α-glucose and β-glucose, two metabolic pathways that are glycolysis or gluconeogenesis, and starch and sucrose metabolism in the liver were markedly altered.

Notoginseng (Sanqi), the root of Panax notoginseng (Burk.) F. H. Chen (Araliaceae), is one of the most valuable traditional Chinese medicines (TCM). It has been widely used in China with a long history for treatment of haemorrhage, edema, and cardiovascular disorders. Steamed P. notoginseng has been considered to have stronger therapeutic functions than raw P. notoginseng in the treatment of tumors, cardiovascular diseases, etc. Saponins are the principal chemical and pharmacological constituents in P. notoginseng. Thus, it is of great importance to determine the constituent saponins and determine any differences between fresh P. notoginseng and steamed P. notoginseng. We used a rapid and direct analytical method based on liquid extraction surface analysis combined with mass spectrometry (LESA-MS) to identify saponins in the xylem, phloem and cambium of fresh and steamed P. notoginseng root slices. The results revealed that ginsenosides Rg1, Rb1, Re, Rd, notoginsenoside R1 and their malonyl group versions were most abundant in fresh root slices, while in steamed slices ginsenosides Rg5, Rk1 and other minor polar components could be detected, and the relative content of large polar components was lower. The described method is fast, robust and sensitive and the process does not need traditional and cumbersome pretreatment such as crushing, extraction and separation. It is the first non-destructive study on the differences in saponins between fresh and steamed P. notoginseng root slices.

The senescence-associated secretory phenotype (SASP) is a generic term for the secretion of a series of cytokines such as pro-inflammatory factors, chemokines and proteases, and is a key feature of senescent cells. SASP is a double-edged sword that can resist a harmful environment in normal cells, but with the decline of body function, the massive secretion of cytokines, chemokines and proteases accelerates aging while inducing inflammation, leading to the development of various aging-related diseases. This article reviews the composition and physiological functions of SASP, the changes in SASP during aging, the regulatory pathways associated with SASP, and the anti-aging drugs that regulate SASP. This article aims to present a more comprehensive understanding of SASP and lay the foundation for SASP-based anti-aging research and the discovery of new targets for anti-SASP drugs.

A new carbazole alkaloid was isolated from the aqueous extract of the stems of Clausena lansium (Lour.) Skeels by various chromatographic methods, including HPD-100, PRP-512A, silica gel, and reverse phase C18. Its structure was determined by spectroscopic and chemical methods, including UV, IR, HR-ESI-MS, 1D/2DNMR and ECD. Compound 1, named as Claulamine F, showed no antimicrobial activity on Staphylococcus aureus, Escherichia coli or Pseudomonas aeruginosa. In addition, compound 1 exhibited no cytotoxicity on five kinds of cancer cells through MTT methods.

The coagulation Ⅷ factor (FⅧ) contains eight pairs of disulfide bonds, which are involved in maintaining its structure and function. It has been demonstrated that the disulfide bond between Cys¹⁸⁹⁹/Cys¹⁹⁰³ of the A3 domain in the light chain impedes secretion. In our previous work, an engineered inter-chain disulfide in the B domain-deleted FⅧ (BDD-FⅧ) promoted heterodimer assembly and secretion of separately expressed heavy and light chains. In this study, we constructed two BDD-FⅧ variants, one of which contains an engineered inter-chain disulfide bond (F8C) between Met⁶⁶² > Cys and Asp¹⁸²⁸ > Cys mutations and another contains an endogenous A3 domain with a disrupted disulfide bond from F8C (F8CG) by replacement of Cys¹⁸⁹⁹ and Cys¹⁹⁰³ with Gly in F8C. We explored their function and secretion. By transducing F8C and F8CG into HEK293 and COS-7 cells, the formation of disulfide bonds and the secretion and coagulation activity of the two variants in the culture media and their binding affinity for von Willebrand factor (vWF) could be observed. The results show that variants F8C and F8CG are mainly the disulfide bonded heavy and light chain dimer, while the wild type BDD-FⅧ (F8) is dominated by the easily dissociated heavy and light chain dimer. The secretion and activity of F8C was significantly higher than that of F8, while the secretion and activity of F8CG was significantly higher than that of F8C. The vWF binding of the two variants is similar to F8. This indicates that the BDD-FⅧ variant F8CG may be attractive molecule for protein replacement and as a transgene in gene-therapy strategies. These findings are encouraging for future studies targeting disulfide bond elimination for further enhancement of FⅧ secretion.

A pre-column derivatization and ultra-high performance liquid chromatography quadrupole time-of-flight mass spectrometry (UHPLC-QTOF-MS/MS) method was developed for qualitative and quantitative determination of medium-and short-chain fatty acids in mice feces, and was further applied to evaluate variations in the feces of mice before and after antibiotic treatment. This animal experiment had been approved by Animal Experimental Ethics Committee of Jiangsu Province Academy of Traditional Chinese Medicine. By optimizing the derivatization conditions and UHPLC-QTOF-MS/MS parameters a new UHPLC-QTOF-MS/MS method with 3-nitrophenylhydrazine as the derivatization reagent was developed for simultaneous determination of 16 medium-and short-chain fatty acids. Validation studies showed that the linearity of the calibration curves was good (R² > 0.99), the RSD of intra-day and inter-day precision was less than 10%, the repeatability RSD was less than 6%, the recovery rate was between 80%-120% at three spiked levels, and the stability RSD was less than 7% within 36 h. The types and amounts of the detected medium-and short-chain fatty acids in feces significantly changed after the mice were treated with antibiotics. The content of formic acid, acetic acid, propionic acid, butyric acid, isobutyric acid, valeric acid, and lactic acid decreased, whereas that of heptanoic acid and succinic acid increased significantly. All these results suggest that the newly established method is accurate and reliable, and can be used for determination of medium-and short-chain fatty acids in feces.

The whole chloroplast genome of the medicinal plant Paeonia mairei H. Lév. was sequenced using the Illumina HiSeq X Ten platform and then assembled, annotated, and characterized by bioinformatic methods in this study. The complete chloroplast genome of P. mairei is 152 731 bp in length with the typical quadripartite structure, which consists of a large single copy-region (LSC, 84 402 bp), a small single copy-region (SSC, 16 969 bp), and a pair of inverted repeat regions (IRa and IRb, 25 680 bp), with an overall GC content of 38.4%. A total of 136 predicted genes, including 90 protein-coding genes, 38 tRNA genes and eight rRNA genes were identified. Among these, seven protein-coding genes, seven tRNA genes and four rRNA genes were found duplicated in the IR regions. In addition, 28 dispersed repeats, 10 tandem repeats, and 64 simple sequence repeats were detected within the whole chloroplast genome of P. mairei. Comparative analyses between 12 Peaonia species showed that the chloroplast genomes are highly conserved in length, gene content, gene order, and GC content. Meanwhile, the noncoding sequences (intergenic regions and introns) show a higher variation than the protein coding sequences, and sequences from the LSC region and SSC region are more variable than those from the IR regions. P. mairei was inferred forming in a distinct clade with P. lactiflora, P. obovate, and P. anomala subsp. veitchii with a 100% bootstrap value and is phylogenetically closest to P. lactiflora. These results may provide a basis for further genetic studies and the development and utilization of medicinal P. mairei.

We compared the pharmacokinetic and pharmacodynamic profiles of desmopressin acetate after intraocular, intravenous and intragastric administration in rabbits to better understand the systemic delivery of peptide drugs through intraocular administration. Fifteen rabbits were randomly divided into three groups (intraocular administration, 7 μg·kg^-1; intravenous administration, 0.7 μg·kg^-1; and intragastric administration, 7 μg·kg^-1). Blood samples were taken from the heart at predetermined time points after dosing and the plasma desmopressin concentration was analyzed by enzyme-linked immunosorbent assay (ELISA). Another 21 rabbits were randomly divided into three groups (intraocular administration, 7 μg·kg^-1; intravenous administration, 0.7 μg·kg^-1; intragastric administration, 7 μg·kg^-1) for a pharmacodynamics study. Urine was collected at predetermined intervals after dosing. The pharmacokinetic parameters after intravenous administration were as follows:C_max was 143.0 pg·mL^-1; the area under the plasma concentration-time curve for desmopressin (AUC_0-t) was 999.9 pg·h·mL^-1. The pharmacokinetic parameters after intraocular administration were as follows:t_max was 5 min, C_max was 125.6 pg·mL^-1, AUC_0-t was 873.1 pg·h·mL^-1, and absolute bioavailability (F) was 8.7%. The pharmacokinetic parameters after intragastric administration were as follows:t_max was 10 min, C_max was 104.1 pg·mL^-1, AUC_0-t was 451.8 pg·h·mL^-1, and absolute bioavailability was 4.5%. Intraocular administration and intravenous administration of one tenth of the dosage showed a similar effect, and the urine volume remained decreased for 12 h, but urine volume increased significantly in the second collection period after intragastric administration, and there was no decrease in volume 12 h after dosing. This study demonstrates that peptide drugs such as desmopressin can be absorbed more rapidly after intraocular administration than after intragastric administration and can exert systemic therapeutic effects. In this study, the program of animal testing had been approved by the Laboratory Animal Care and Use Committee at Anhui University of Chinese Medicine.

3-Hydroxy-3-methylglutaryl coenzyme A reductase (HMGR) is the first rate-limiting enzyme of terpenoid biosynthesis in the mevalonic acid pathway (MVA) pathway. It is an important regulatory site in terpenoids metabolism pathway in the cytoplasm. According to the transcriptome database of Cinnamomum camphora, two HMGRs named CcHMGR1 (GenBank:MN163055) and CcHMGR2 (GenBank:MN163056) were cloned by cDNA from C. camphora. The ORF of CcHMGR1 and CcHMGR2 is composed of 1 689 bp and 1 683 bp, respectively, encoding 562 and 560 amino acids. The bioinformatics analysis of CcHMGR1 and CcHMGR2 indicated that the molecular weight of the encoded protein is 59.819 kDa and 59.397 kDa, with a theoretically isoelectric point of 8.20 and 8.61, respectively. There are 2 transmembrane structures without signal peptide existing in the encoded amino acid of CcHMGRs. The analysis of sequence alignment and phylogenetic tree showed that the CcHMGRs belonged to the HMGR family. The camphor is divided into five chemitypes, according to the main chemical compounds in C. camphora. The results of the real time PCR indicated that the expression level of CcHMGRs in Cineol type was higher than that in Linalool type, iso-nerolidol type, Camphor type and Borneol type. CcHMGRs expressed highest in roots and lowest in branches. In this study, the cDNA full length of CcHMGRs were cloned from C. camphora for the first time. Our results revealed that the expression level of CcHMGRs were different among five chemical types and different plant tissues, and the research provides foundation for further study of the terpenoids biosynthetic pathway in C. camphora.