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Background Immune checkpoint molecules, especially programmed death 1 (PD-1) and its ligand, programmed death ligand 1 (PD-L1), protect tumor cells from T cell-mediated killing. Immune checkpoint inhibitors, designed to restore the antitumor immunosurveillance, have exhibited significant clinical benefits for patients with certain cancer types. Nevertheless, the relatively low response rate and acquisition of resistance greatly limit their clinical applications. A deeper understanding of the regulatory mechanisms of PD-L1 protein expression and activity will help to develop more effective therapeutic strategies.Methods The effects of AR-A014418 and THZ531 on PD-L1 expression were detected by western blot, reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and flow cytometry. In vitro kinase assays with recombinant proteins were performed to confirm that AR-A014418 functioned as a CDK12 and CDK13 dual inhibitor. The roles of CDK12 and CDK13 in intronic polyadenylation (IPA) and transcription of PD-L1 were determined via RNA interference or protein overexpression. T-cell cytotoxicity assays were used to validate the activation of antitumor immunity by AR-A014418 and THZ531.Results AR-A014418 inhibits CDK12 to enhance the IPA, and inhibits CDK13 to repress the transcription of PD-L1. IPA generates a secreted PD-L1 isoform (PD-L1-v4). The extent of IPA was not enough to reduce full-length PD-L1 expression obviously. Only the superposition of enhancing IPA and repressing transcription (dual inhibition of CDK12 and CDK13) dramatically suppresses full-length PD-L1 induction by interferon-γ. AR-A014418 and THZ531 could potentiate T-cell cytotoxicity against tumor cells.Conclusions Our work identifies a new regulatory pathway for PD-L1 expression and discovers CDK12 and CDK13 as promising drug targets for immune modulation and combined therapeutic strategies.

Abstract Background This study aimed to identify genes related to the degree of immune cell infiltration in head and neck squamous cell carcinoma (HNSCC), explore their new biological functions, and evaluate their diagnostic and prognostic value in HNSCC. Methods Transcriptomic data from The Cancer Genome Atlas (TCGA) HNSCC dataset was used to screen differentially expressed genes between tumors and normal tissues, followed by weighted correlation network analysis (WGCNA) to identify immune‐related modules. Differential gene expression, immune cell infiltration, and survival analyses were performed to screen key genes. The expression of these key genes was validated in Oncomine and gene expression omnibus (GEO) datasets and by immunohistochemistry (IHC). Results 1869 and 1578 genes were significantly upregulated and downregulated in HNSCC. WGCNA showed that the brown module was associated with the most significant number of immune‐related genes. PPI network analysis demonstrated that PPL, SCEL, KRT4, KRT24, KRT78, KRT13, SPRR3, TGM3, CRCT1, and CRNN were key components in the brown module. Furthermore, the expression levels of KRT4, KRT78, KRT13, and SPRR3 in HNSCC correlated with infiltration levels of CD8+ T cells and macrophages. Survival analyses revealed that the expression of KRT78, KRT13, and SPRR3 in HNSCC correlated with overall survival (OS). The IHC assay indicated that KRT13 (p = .042), KRT78 (p

[Objectives] To determine the optimum conditions and antioxidant aclivily of Myrica ruba leaf pigment, and provide reference for further development and use of Myrica ruba leaves. Methods] Pigment of Myrica rulxi leaves was extracted by ethanol extraction method. Ab-sorbance was measured at wavelength of 3(H) nra. Effects of ethanol volume fraction, solid-to-liquid ratio, temperature, and extraction lime were studied. L9(33) orthogonal experiment was carried out to optimize the extraction process, and DPPH scavenging ability was measured. Results] The optimum extraction process of Myrica ruba leaf pigment was as follows; the ethanol volume fraction was 50% ; the solid-to-liq-uid ratio was 1 ;60 ( w/v), extraction temperature °C; extraction time was 60 min; after the concentration of purified Myrica ruba leaf pigment reached 0.4455 mg/mL. the DPPH scavenging ability was basically the same as the same concentration Vc. Conclusions] The experiment indicates that the optimized extraction process has a high yield rale, and Myrica rulxi leaf pigment has high anli-oxidant activity. [ABSTRACT FROM AUTHOR]

[Objectives] To optimize the extraction process of Herba Lycopi polysaccharide by cellulase, and study its antioxidant activity. [Methods] The singe factor test and orthogonal test were conducted to study the effect of pH, enzyme dosage, enzymolysis temperature and extraction time on the extraction of Herba Lycopi polysaccharide. And the test was done about its scavenging effect on DPPH radicals and superoxide anions, and the antioxidant activity was evaluated. [Results] The optimum extraction process by ceHulase method was as foUows : pH value of 7; enzyme dosage of 4 ml; enzymolysis temperature of 40 °C; extraction time of 2.5 h. The average extraction rate of Herba Lycopi polysaccharide was 10. 70%, and RSD was 0.17%. The antioxidant activity test showed that Herba Lycopi polysaccharide had a significant scavenging effect on radicals. [Conclusions] Herba Lycopi polysaccharides had strong antioxidant activity, and could provide a scientific basis for the development and utilization of Herba Lycopi medicinal resources. [ABSTRACT FROM AUTHOR]

Traditional extraction methods of total saponins of Panax notoginseng include cold soaking method, water decoction method, alcohol reflux method, percolation method, macroporous resin adsorption method, and accelerated solvent extraction (ASE) method. Modern extraction methods include ultrasonic extraction, microwave asisted extraction, supercritical C02 extraction, microbial fermentation assisted extraction, neural network model optimized extraction method, and multi-stage countercurrent extraction method. This paper discussed principles of these methods and compared their advantages and disadvantages. [ABSTRACT FROM AUTHOR]

[Objectives] To compare different extraction methods for total alkaloids from Abrus Cantoniensis Hance taking chloroform a solvent. [Methods] The ethanol extraction method, ultrasonic extraction method, acid water extraction method, and Soxhlet extraction method were used to extract total alkaloids, and the total alkaloids of Abrus Cantoniensis Hance were measured at 238 nm wavelength. [Results] The extraction rate of ethanol extraction method was 14. 76 mg/100 g; the extraction rate of ultrasonic extraction method was 56. 85 mg/100 g; the extraction rate of acid water extraction method was 56. 63 mg/100 g; the extraction rate of Soxhlet extraction method was 58. 19 mg/100 g. [Conclusions] Results indicate that Soxhlet extraction method can obtain the highest extraction rate. [ABSTRACT FROM AUTHOR]