In order to explore the genetic diversity and structure of Bupleurum chinense, we used 18 pairs of SSR molecular markers to analyze the genetic diversity of 619 individuals in 62 cultivated and wild populations of Bupleurum chinense from Shanxi and the surrounding provinces. The results show that the 62 Bupleurum chinense populations have high genetic diversity, with that of the wild Bupleurum chinense populations greater than that of cultivated populations. AMOVA analysis indicated that genetic variation within populations was greater than between populations. Principal coordinate analysis (PCoA) divided the Bupleurum chinense populations into 3 groups, the first group containing wild Bupleurum chinense populations from all parts of Shanxi, the second group consisting of cultivated Bupleurum chinense populations from Shanxi, Hebei, Shaanxi and Liaoning, and the third group consisting of cultivated Bupleurum chinense from Shanxi and Gansu. STRUCTURE software cluster genetic structure analysis grouped the 62 Bupleurum chinense into two populations: the first group composition was the same as the population classified as the third category in the PCoA analysis, while the second group includes the populations from the first and second categories of the PCoA. PCoA, cluster genetic structure analysis, and NJ tree cluster all gather wild Bupleurum chinense population into a single category, distinguishing it from the cultivated populations. This study provides a theoretical basis for the utilization of germplasm resources, genetic variation and the development of quality germplasm resources for Bupleurum chinense.

This study investigated the effect of morin, an active ingredient of the family Moraceae, on collagen-induced arthritis (CIA) in mice, and explored the underlying mechanism from the perspective of recovering immune balance. The collagen was used to induce model of CIA in mice, morin was administered by gavage, and arthritis index (AI) score, imaging and histopathological changes of the paws and ankle joints, and the levels of proinflammatory factors, proinflammatory mediators as well as the IgG class antibodies in serum were detected. In addition, the frequencies of T helper 17 (Th17) and regulatory T (Treg) cells and the levels of relevant transcription factors and functional factors in lymph nodes/spleen as well as the levels of interleukin-17A (IL-17A) and IL-10 in serum were determined. The results showed that oral administration of morin significantly reduced the AI score, improved joint swelling and bone damage, reduced the pathological score, and down-regulated the levels of proinflammatory factors [tumor necrosis factor-‍α (TNF-‍α), IL-6 and IL-1β], proinflammatory mediators [prostaglandin E₂ (PGE₂), matrix metalloproteinase-13 (MMP-13) and nitric oxide (NO)] and IgG class antibodies (IgG and IgG2a) in serum. Moreover, the percentage of Th17 cells, the expressions of Th17-specific transcription factor retinoic acid-related orphan receptor γt (RORγt) and functional factors IL-17A, IL-21 and IL-22 in lymph nodes/spleen, as well as the level of IL-17A in serum were down-regulated, while the percentage of Treg cells, the expressions of Treg-specific transcription factor forkhead box P3 (Foxp3) and functional factors IL-10 and transforming growth factor-‍β (TGF-‍β) in lymph nodes/spleen, as well as the level of IL-10 in serum were up-regulated. All animal treatments were approved by the Animal Ethics Committee of China Pharmaceutical University and strictly followed the welfare regulations of laboratory animals of China Pharmaceutical University. This study indicates the therapeutic effect of morin on mice with CIA, and the mechanism is associated with the improvement of Th17/Treg imbalance, which provides a theoretical basis for the clinical application of morin.

α3β4 nicotinic acetylcholine receptors (nAChRs) are potential therapeutic targets in diseases such as addiction, cancer, and obesity. In this study, by replacing three amino acids of the α3 subunit with the corresponding positions of the rα6 subunit simultaneously, an α3[K152E, E184D, Q195T] subunit mutant was constructed by PCR-mediated site-directed mutagenesis and its cRNA was obtained by in vitro transcription. The cRNA of mutant subunits mixed in equal molar ratios with β4 subunits were microinjected into Xenopus oocytes. The pharmacological activity and function of α3[K152E, E184D, Q195T]β4 nAChR was evaluated by a two-electrode voltage clamp electrophysiological technique. Acetylcholine, nicotine, and cytisine were used as agonists to evaluate the magnitude of ligand-gated currents and gating characteristics of wild-type and mutant α3β4 nAChRs. The half-maximal effective concentrations (EC₅₀) of acetylcholine, nicotine, and cytisine on wild-type α3β4 nAChRs were 277.5, 34.02 and 23.05 µmol·L^-1, respectively, while their EC₅₀ values with α3[K152E, E184D, Q195T]β4 nAChR were 170.5, 26.6, and 98.45 µmol·L^-1, respectively. Thus these EC₅₀ values for the three agonists towards the mutant receptor were changed 0.6-fold, 0.8-fold, and 4.3-fold, respectively, compared with the wild-type receptor; cytisine was most strongly affected, with a 77% decrease in potency. However, the maximum agonistic efficiency (E_max) of cytisine on wild-type and mutant α3β4 nAChRs was increased from 94.12% to 155.08% relative to the peak current amplitude induced by 1 mmol·L^-1 acetylcholine. Thus, although the α3[K152E, E184D, Q195T]β4 nAChR had significantly reduced sensitivity to cytisine, the maximum current amplitude induced by cytisine was clearly increased. This mutant had slightly increased sensitivity to acetylcholine and nicotine. The results indicate that these three amino acids of the α3 subunit have important and varying effects on ligand binding to the α3β4 nAChR, providing a basis for further structure-functional research on α3β4 nAChR, as well as the pathology of related diseases.

Using Lonicera japonica genomic DNA as a template, we cloned Lonicera japonica U6 promoters. Four LjU6 promoters, 336, 708, 359 and 602 bp in length, were cloned by PCR from Lonicera japonica genomic DNA. PlantCARE analysis found that the four promoters contained typical promoter cis-elements, such as a TATA box and CAAT box, and contained regulatory elements related to light response and stress response. After the cloning products were sequenced, the LjU6 promoter was ligated to the pBI121 vector carrying the β-glucuronidase (GUS) gene to construct four LjU6-pBI121 fusion expression vectors. Nicotiana tabacum leaves were transformed by the Agrobacterium transient transformation method and GUS histochemical staining was performed on the leaves. The staining results showed that LjU61-F1 had the highest transcriptional activity. This study thus identified a U6 promoter with high transcriptional activity, providing a basis for the establishment of CRISPR/Cas9 genome editing technology in Lonicera japonica.

The purpose of the study was to investigate the thermal expansion characteristics of brivaracetam form Ⅰ, and explore the influence mechanism of the crystal structure on its thermal expansion behavior. The crystal structure of brivaracetam form Ⅰ was characterized by X-ray single crystal diffraction and variable temperature X-ray powder diffraction at different temperatures. The interaction energy of brivaracetam molecule calculated by B3LYP/6-31G(d, p) wave function with the aid of CrystalExplorer 21.5 software. The results show that brivaracetam form Ⅰ exhibits significant reversible anisotropic thermal expansion under the temperature range of 123-323 K. The principal expansion X₁, X₂, X₃ axes are approximately aligned with the a, b and c axes of the unit cell, and the thermal expansion coefficients of the principal expansion axes are -127.61×10^-6, 95.96×10^-6, 233.80×10^-6 K^-1, respectively. The a-axis exhibits negative expansion characteristics. The volumetric thermal expansion coefficient is 202.17×10^-6 K^-1. The energy framework of the crystal is obvious layered, and the interaction energy between layers is weak, which leads to a significant linear positive expansion in the c-axis direction of the unit cell. Through a combination of experimental and theoretical methods, the thermal expansion characteristics of brivaracetam form Ⅰ are systematically analyzed, and the influence mechanism of the crystal structure on its thermal expansion behavior is explored, which has certain guiding significance for the production process of tablet preparations in practice.

Lornoxicam (LOR) is a nonsteroidal anti-inflammatory drug with analgesic, anti-inflammatory and antipyretic effects. As a biopharmaceutics classification system (BCS) class Ⅱ drug, it has poor aqueous solubility and then low bioavailability after oral administration. In addition, the tabletability of LOR itself is also poor and could not form the tablet after compression, which seriously limits the development of its oral solid dosage. The current study aims to improve dissolution and tabletability of LOR by cocrystallization technique with small molecule puerarin (PUE). LOR cocrystal with the co-former PUE was prepared via the solvent-evaporation method and characterized by powder X-ray diffraction, differential scanning calorimetry, Fourier transform infrared spectroscopy and thermo-gravimetric analyzer. The dissolution behavior, tabletability and stability of the prepared cocrystal were also further investigated. In comparison to pure LOR, LOR-PUE cocrystal showed higher apparent and intrinsic dissolution rate. Moreover, after cocrystallization, the solubility of LOR and PUE showed 4.0-fold and 1.5-fold increase compared to the raw ones in water, respectively. LOR-PUE cocrystal showed significantly improved tabletability compared to LOR alone under a wide compression range of 75-375 MPa. In addition, such cocrystal exhibited superior chemical stability with no change of drug contents for at least 60 days under the conditions of 40 ℃ and 25 ℃/75% RH.

To expand the structural diversity of Matijin-Su (MTS) derivatives and explore novel anti-HBV activity compounds, a series of fluorinated dipeptidomimetics of MTS were designed and synthesized by using trifluoromethyl substituted methylamine unit as bioisostere to replace the amide bond of the MTS derivatives. The structures of all target compounds were confirmed by ¹H NMR, ¹³C NMR, ¹⁹F NMR, HRMS, or ESI-MS, and the crystal structure of 10' was determined by X-ray single crystal diffraction. Their inhibitory activity against hepatitis B virus (HBV) in vitro were evaluated using HepG2 2.2.15 cell model. The results showed that all target compounds had inhibitory effect on HBV DNA replication, the IC₅₀ of 14e, 14f, and 14k were 0.37, 0.29, and 0.79 μmol·L^-1, respectively.

Histone deacetylases (HDACs) are a class of key enzymes that regulate epigenetics. There are 5 small-molecule HDACs inhibitors having been approved for anti-cancer therapy on the market. In recent years, there have been more and more studies on the antiviral aspects of HDACs inhibitors. This article classifies viruses into human immunodeficiency virus 1 (HIV-1), new coronavirus (SARS-CoV-2), Epstein-Barr virus (EBV) and other viruses, systematically summarizes the recent advances of antiviral effects of the HDACs inhibitors from the perspective of medicinal chemistry. This review aims to provide the researchers the convenience of accessing the latest advances of the antiviral effects of HDACs inhibitors, and to analyze the challenges and prospects of this field in future drug discovery.

The rational medication in pregnant women is a clinical issue that clinicians and pharmacists must take seriously. Most tissues and organs undergo anatomical and physiological changes during pregnancy that affect the absorption, distribution, metabolism, and excretion of drugs in vivo, which ultimately lead to changes in bioavailability. In order to achieve an effective therapeutic concentration, dose adjustment might be required during this period. In the past ten years, the application of modeling and simulation methods in the field of drug development and clinical therapy has continued to expand, for instance, using population pharmacokinetic (PPK) and physiologically based pharmacokinetic (PBPK) modeling to adjust dosage regimen in special populations. Rigorously designed and validated models will effectively make up for the deficiencies of clinical trials, provide valuable references for the design of clinical research, and even replace part of them. This article will introduce the physiological changes that affect the pharmacokinetic properties of the drug during pregnancy and review the progress in the application of PBPK modeling in pharmacokinetic studies in pregnant women.

We analyzed the anticancer effect and mechanism of the novel indoleamine 2, ‍3-dioxygenase 1 (IDO1) inhibitor NLG-919 combined with temozolomide (TMZ) on human glioma cell lines. The anti-tumor activity of NLG-919 and temozolomide after single and combined treatments was detected by MTT assay. Colony formation assay, invasion assay and migration assays were used to detect the effects of NLG-919 and temozolomide alone or in combination on proliferation, invasion and migration of human glioma cells. A flow cytometry assay was used to detect cell apoptosis, cell cycle arrest, reactive oxygen species (ROS) production and mitochondrial membrane potential damage (JC-1). An immunofluorescence assay was used to detect the expression level of IDO1 and HPLC was used to detect the expression level of L-kynurenine (Kyn) to explore the anti-tumor mechanism of NLG-919 and temozolomide. The results show that NLG-919 had a weak in vitro inhibitory effect compared to that of temozolomide. The IC₅₀ of NLG-919 on U251 cells and U87 after 72 h was 26.9 and 30.7 μmol·L^-1, respectively. However, when NLG-919 was used in combination with temozolomide, its anti-glioma activity was significantly increased. Compared with the single treatment, the combination treatment had a potent ability to inhibit proliferation, invasion and migration of glioma cells. Combination treatment improved the capacity of temozolomide to induce cell cycle arrest and inhibit the growth of glioma cells. NLG-919 significantly down-regulated the expression and activity of IDO1 in glioma cells, and the inhibitory effect was improved after combination with temozolomide, and effectively blocked the production of Kyn through the metabolism of L-tryptophan (Trp). In conclusion, the IDO1 inhibitor NLG-919 and temozolomide showed synergistic effects in the anticancer therapy of human glioma cell lines.