G protein-coupled receptors (GPCRs), the largest family of transmembrane receptors in the human, contain seven transmembrane helices, and are usually regarded as critical drug targets because of their key roles in multiple diseases. Currently, 30%-40% approved drugs target GPCRs. Nanobodies (also known as single domain antibodies) are important research tools for GPCRs due to their small molecular weight, good biochemical properties and high affinity for "cracks or cavities". In addition, nanobodies have long complementarity determining region 3 (CDR3) loops which can be inserted deeply into GPCRs ligand binding pockets, efficiently binding to the folds. This review summarizes the characteristics of nanobodies and their applications in GPCRs research and briefly introduces the current identification routes of targeted GPCRs nanobodies, which could provide new idea and method for applications of nanobodies in GPCRs research.

Based on the chemical structure of known compound, 12 isatin derivatives palmitoyl transferase inhibitors are designed and synthesized using bioisosterism and molecular docking, while their anti-tumor activities in vitro are determined. The structures of the target compounds are confirmed by ¹H NMR, ¹³C NMR and HR-MS. In vitro anti-tumor assay illustrates that compound 5b exhibits similar anti-tumor activity to the control (IC₅₀ = 8.4 μmol·L^-1), with IC₅₀ value of 12.0 μmol·L^-1 against MCF-7 in which palmitoyl transferase is highly expressed. Compound 4b shows higher inhibitory activity against HeLa (IC₅₀ = 8.1 μmol·L^-1) than cisplatin (IC₅₀ = 40.1 μmol·L^-1). The molecular docking demonstrates that all compounds could completely enter the site of 3'-adenosine monophosphate-5'-diphosphate (PAP). Taken together, isatin derivatives represent promising compounds for the discovery of novel anti-tumor agents.

In this study, a novel nano-drug delivery system, namely hybrid exosome, was constructed via membrane self-assembly of pancreatic cancer cell-derived exosomes with liposomes, which inherits the functionalities of exosomes, including high affinity, good stability and low immunogenicity, but also unites the characteristic of liposomes (e.g., long circulation time, high loading ability) to achieve precise drug navigation and minimum adverse effects. Specifically, two different preparation methods—repeated freeze-thawing and 37 ℃ incubation were used to fabricate hybrid exosomes at laboratory scale. Comparative analysis and characterization of these synthesized samples were performed based upon size, zeta potential and membrane fusion efficiency. The results showed that the highest exosome yield was attainted after culture for 48 h, with the exosome yield of 0.83 ± 0.07 mg/10⁸ cells for HuP-T3 cell line and 0.79 ± 0.10 mg/10⁸ cells for Panc0403 cell line. Hybrid exosomes obtained by freeze-thaw method were shown to have higher membrane fusion rate, lower size and polydispersity index (PDI), higher zeta potential and relative more stable, as compared with that made by incubation at 37 ℃ for 12 h, indicating the former approach is more suitable to construct hybrid exosomes with desirable physicochemical properties. This result may provide a preliminary experimental basis for the subsequent delivery of different anticancer drugs for the treatment of solid tumors such as pancreatic cancer.

To explore the protective effect of protropine in Corydalis humosa Migo. on lipopolysaccharide-induced acute kidney injury in mice (AKI), an approach that used ultra-performance liquid chromatography quadrupole time-of-flight tandem mass spectrometry (UHPLC-Q/TOF-MS) coupled with a multivariate analytical platform was established. The BALB/c mice were divided into normal group (CON), model group (LPS), and protropine group (PRO). Mice were injected intraperitoneally with lipopolysaccharide solution to replicate the AKI model. Three hours after modeling, mice were given the protropine solution by gavage. Protropine was a monomer compound isolated in the laboratory, and protropine solution was prepared by dissolving protropine in sterilized distilled water. Administration was performed twice a day for three days. After modeling and administration, serum samples were collected. UHPLC-Q/TOF-MS was used to generate metabolomics data. Multivariate statistical analysis and online databases were used to screen potential biomarkers and enrich metabolic pathways. The heatmap of relative quantitative biomarker data was generated through Mev software. Animal experiments were approved by the Animal Experimentation Ethics Committee of Henan University of Chinese Medicine (No. SYXK2015-0005). The results show that the metabolic profile of mice in the LPS group was significantly altered by intervention with protropine, and clustered towards the CON group. 70 biomarkers were identified from the CON group vs LPS group (35 in positive source mode, 35 in negative source mode), and 67 biomarkers were identified from the LPS group vs PRO group (37 in positive source mode, 30 in negative source mode). A total of 34 common markers (18 in positive source mode, 16 in negative source mode) were obtained from the two comparison groups. The enrichment of all biomarkers resulted in 8 metabolic pathways including linoleic acid metabolism, D-glutamine and D-glutamate metabolism, arginine and proline metabolism, and arachidonic acid metabolism. The results show that protropine in Corydalis rhizoma ameliorates the kidney damage, insufficient energy supply, and inflammation in AKI mice by regulating amino acid metabolism, energy metabolism, and lipid metabolism in AKI mice.

Baihu-Guizhi Decoction (BHGZD), a prescription from ''Synopsis of the Golden Chamber'', has a definite clinical effect in the treatment of rheumatoid arthritis (RA). However, the research on the mechanism of this prescription mainly focuses on the regulation of inflammatory response and immune function, and its efficacy and mechanism of inhibiting synovial angiogenesis have not been reported. In the current study, transcriptomics data mining, biological network analysis and ''in vivo-in vitro'' experimental verification integrated research strategy to explore the potential and molecular mechanism of BHGZD in RA synovial angiogenesis with hot syndrome. Animal welfare and experimental procedures follow the regulations of the Animal Ethics Committee of China Academy of Chinese Medical Sciences. The results of network analysis showed that the candidate network targets of BHGZD intervention in RA with hot syndrome were significantly involved in multiple angiogenesis regulation related pathways. Among them, vascular endothelial growth factor A-vascular endothelial growth factor receptor 2 (VEGFA-VEGFR2) signaling pathway contains multiple BHGZD candidate network targets, such as VEGF, phosphatidylinositol 3-kinase (PI3K), protein kinase B (AKT), etc. Further experimental results showed that BHGZD could effectively reduce the expression of CD31 in knee synovium, the expression level of VEGF in serum, the activity of endothelial nitric oxide synthase (eNOS), phosphorylated VEGFR2 (p-VEGFR2), p-PI3K and p-AKT in joint tissue of adjuvant-induced arthritis rats with hot syndromes, the migration and invasion activity of HUVEC and MH7A cells, and the lumen formation activity of HUVEC cells and improve the expression level of endostatin in serum. In conclusion, BHGZD has the potential to alleviate excessive synovial angiogenesis in RA with hot syndrome, and its mechanism may be related to the intervention of VEGF/VEGFR2/PI3K/AKT signaling pathway.

The development of the manufacturing process may require considerable time and resources from an economic perspective, which may result from the lack of cost-effective and reliable modeling tools of unit operation development in the pharmaceutical industry, in contrast to other chemical industries. Therefore, it is necessary to apply the modeling tools to the process, not only to overcome the challenges of regulatory and economic aspects but also to develop a more efficient and robust process. In response to this necessity, the modeling of the manufacturing process has been become increasingly important, as it can be applied to equipment design, improving process efficient, scale-up and unit operation development in the pharmaceutical industry. Discrete element method is a numerical method for predicting mechanical dynamics, such as position, velocity and motion of individual particles. First of all, the input parameters related to particle contact should be clearly defined. In this work, a calibration method of discrete element parameters was established and then elucidated the effects of different testing methods on repose angle of microcrystalline cellulose (MCC), from mesoscale angle. This experiment was composed of three parts: ① Angle of repose measured by the lifting cylinder method (θ) was regarded as the response value of the model, and then discrete element simulation parameters were screened and optimized by Plackett-Burman, steepest climb and Box-Behnken test designs; ② The robustness of previous model was assessed by angle of repose measured by the funnel injection method (α) and the shear box method (φ) to obtain the best parameter combination generated from the model; ③ Based on accurate and reliable microscopic parameters, the formation mechanism of angle of repose was comprehensively investigated from the mesoscopic-angle perspective. The calibration results showed a robust and reliable parameter combination. Moreover, the lifting speed of lifting cylinder method and the height of funnel injection method all had a certain impact on the measurement results of angle of repose. Interesting, the evolution of force chains in the process of stacking with different angle of repose revealed a certain law in the perspective of mesoscopic-angle. Thus, the objective of present work is to provide a reference for discrete element simulation parameter calibration of other solid preparations and accurate simulation of materials in the pharmaceutical process such as mixing, transferring and tablet pressing.

Inflammatory bowel disease (IBD) is a disease characterized by chronic and progressive inflammation of the intestinal tract, which seriously affects the quality of life of patients because it is difficult to cure and easy to recur. As the main organ of substance absorption, the changes of intestinal drug transporters will lead to changes in the behavior of endogenous and exogenous substances in vivo. Changes in the expression and function of a variety of drug transporters have also been observed in intestinal inflammatory tissues of patients with IBD. This paper reviews the changes of intestinal drug transporters in IBD and its related mechanisms, which provides a theoretical basis for finding new strategies for the treatment of IBD and clinical rational drug use.

Pyroptosis is a form of inflammatory programmed cell death pathway. In vitro and in vivo studies have shown that pyroptosis contributes to the development of macrovascular complications of diabetes, mainly through activating inflammasomes and caspase-1/4/5/11, cleaving gasdermin D (GSDMD), releasing interleukin-18 (IL-18), IL-1β and other inflammatory cytokines. In recent years, the effect of systemic chronic inflammation caused by pyroptosis through inflammatory cascade reaction on macrovascular complications of diabetes has received long-term attention. This article reviews studies of pyroptosis in macrovascular complications of diabetes and the related drugs to provide promising thought for treating macrovascular complications of diabetes in clinical.

Coronavirus disease 2019 (COVID-19) continues to be prevalent all over the world and mutant strains are constantly appearing, the application of vaccine is still an important method of epidemic prevention and control. Mucosal immunity plays an important role in preventing severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) invasion. The currently marketed injectable COVID-19 vaccine mainly activates humoral immunity, but it is difficult to induce effective mucosal immunity, and it is unable to prevent pathogen invasion in the early stage of virus infection. Compared with injection vaccination, inoculation of the COVID-19 vaccine through mucosal routes such as nasal or oral can closely imitate the natural infection pathway of the virus and induce a comprehensive immune response. It is an ideal choice for rapid and extensive vaccination because it has the advantages of simple and convenient use, easy to achieve self-management of vaccinators, reduced demand for professional medical personnel and so on. In this paper, we summarized and analyzed the products and technical platforms of COVID-19 vaccine inoculated by oral route, in order to provide reference for related research work.

Liver fibrosis is characterized by scarring of liver tissue, which is an intermediate pathological process of chronic liver disease developing into liver cancer. Its mechanism involves multiple signal pathways, and its reversibility is a current research hotspot. Bone marrow mesenchymal stem cells (BMSCs) are adult stem cells with multi-differentiation potential. They have ability to differentiate into liver-like cells in vivo and in vitro to perform normal liver cell functions. Modern pharmacological experimental studies have shown that the use of BMSCs alone or in combination with active factors, Chinese medicine or Chinese medicine monomers, genetic modification and other methods can promote their proliferation, differentiation, and migration, improve the therapeutic effect, and play a role in improving liver fibrosis. By summarizing the existing literature, the therapeutic mechanism of BMSCs in improving liver fibrosis is reviewed from the aspects of the pathogenesis of liver fibrosis, the improvement mechanism of liver fibrosis, the biological characteristics of BMSCs and its improvement mechanism, so as to provide reference for the later development of BMSCs cell therapy.