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Objective::To study the effect of Yupingfeng granule on the degranulation of skin mast cells in chronic urticaria (CU) rats and the intervention mechanism of interleukin-23(IL-23), interleukin-17(IL-17) inflammation axis. Method::Totally 60 SPF SD rats were selected and randomly divided into normal group (normal saline), model group (normal saline), and loratadine group (0.9 mg·kg-1·d-1), high-dose Yupingfeng granules group (4.05 g·kg-1·d-1), middle-dose group (2.7 g·kg-1·d-1), low-dose group (1.35 g·kg-1·d-1). The CU rat model was reproduced through intraperitoneal injection of ovalbumin with aluminum hydroxide suspension and DTP vaccine. Histopathological changes of rat skin were observed by hematoxylin-eosin (HE) staining. Degranulation of mast cells in rat skin was determined by toluidine blue staining. IL-23 and IL-17 protein expressions in skin tissue were determined by immunohistochemistry (IHC). IL-23 and IL-17 mRNA transcription levels in skin tissue were determined by real-time fluorescence quantitative polymerase chain reaction (Real-time PCR). Result::Yupingfeng granules can significantly alleviate the pathological manifestations of dermal edema, collagen beam distance, inflammatory cell infiltration of CU rats, and reduce the degranulation reaction of skin tissue mast cells in CU rats. The IL-23, IL-17 mRNA and protein expressions of the skin of model group were significantly increased compared with the normal group (P<0.05, P<0.01). Compared with the model group, Yupingfeng granules can significantly down-regulate IL-23 mRNA and protein expressions of CU rats (P<0.05, P<0.01). Yupingfeng granules had no significant regulatory effect on IL-17. Conclusion::Yupingfeng granule can significantly reduce the degranulation of mast cells in skin tissue of CU rats, and improve the pathological manifestations, such as dermal edema, serous exudation and inflammatory cell infiltration. The mechanism may be related to inhibiting the secretion of IL-23 pro-inflammatory cytokines and improving CU lesions.
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Objective:To investigate the effect of Danggui Yinzi on allergic reaction in chronic urticaria (CU) mice model and the mechanism of autophagy intervention. Method:The SPF BALB/c mice were used to replicate the CU mice model by intraperitoneal injection of ovalbumin and aluminum hydroxide suspension. The animals were randomly allocated into six groups: a normal group (normal saline 20 mL·kg-1·d-1), a model group (normal saline 20 mL·kg-1·d-1), a loratadine group(0.001 3 g·kg-1·d-1), a Danggui Yinzi high，medium and low-dose group（39.3，19.6，9.8 g·kg-1·d-1）. The pathological changes of skin tissues were observed by hematoxylin-eosin (HE) staining. Morphological changes of autophagy in skin tissues epithelial cells were observed by transmission electron microscope. The mRNA levels of microtubule-associated protein 1 light chain 3B(LC3B) and ubiquitin-binding protein p62 mRNA in skin tissues were detected by real-time quantitative polymerase chain reaction (Real-time PCR). The expressions of LC3B and p62 in skin tissues were detected by immunohistochemistry (IHC). Result:Danggui Yinzi can significantly improve the pathological manifestations of dermal edema, collagen bundles separation, telangiectasia in CU mice， it can also improve autophagosomes formation and abnormal cell ultrastructure such as nuclear chromatin condensation, mitochondrial swelling, endoplasmic reticulum expansion, etc. Compared with the normal group, the protein expressions of LC3B in skin tissues of the model group was significantly increased (P<0.01), LC3B mRNA level was increased too, while p62 mRNA levels and its protein expressions were decreased-regulated (P<0.01). Compared with the model group, levels of LC3B mRNA and protein expressions of the Danggui Yinzi groups were significantly increased (P<0.05,P<0.01), while p62 mRNA levels and its protein expressions were significantly decreased-regulated (P<0.05,P<0.01). Conclusion:Danggui Yinzi can regulate the expression of LC3B, p62 mRNA and protein expressions, enhance the level of autophagy, and improve the pathological state of CU mice.
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@#嵌合抗原受体T（chimeric antigen receptor T, CAR-T）细胞是通过基因工程技术将T细胞改造成针对肿瘤特异性抗原 的新型杀伤细胞，具有特异性强、效率高、 非MHC限制等优点，在复发/难治性血液系统肿瘤和部分实体瘤中取得良好的治疗效 果。但目前CAR-T细胞治疗仍面临着缺乏“现货供应”的通用型CAR-T细胞、抑制性免疫微环境和T细胞耗竭等问题。近年来， 锌指核酸酶(zinc finger nucleases, ZFNs)、转录激活子样效应因子核酸酶(transcription activator like effector nucleases,TALENs)、 规 律性重复短回文序列簇[clustered regularly interspaced short palindromic repeats/CRISPR-associated(Cas9)，CRISPR/Cas9]等新型基 因编辑技术被广泛应用于细胞免疫治疗，为解决上述问题带来了希望。本文综述了目前CAR-T细胞治疗的研究进展及存在问 题，并探讨了3种主要的基因编辑技术改良CAR-T细胞治疗的策略， 为CAR-T细胞的基础研究和临床治疗提供参考。
ABSTRACT
DNA repair processes play a role in the development of drug resistance which represents a huge obstacle to leukemia chemotherapy.Histone H2AX phosphorylation (ser139) (γH2AX) occurs rapidly at the onset of DNA double strand break (DSB) and is critical to the regulation of DSB repair.If DNA repair is successful,cells exposed to anti-neoplastic drugs will keep entering the cycle and develop resistance to the drugs.In this study,we investigated whether γH2AX can be used as an indicator of tumor chemosensitivity and a potential target for enhancing chemotherapy.K562 and multi-drug resistant cell line K562/A02 were exposed to adriamycin (ADR) and γH2AX formed.Flow cytometry revealed that percentage of cells expressing γH2AX was increased in a dose-dependent manner and the percentage of K562/A02 cells was lower than that of K562 cells when treated with the same concentration of ADR.In order to test the potential of γH2AX to reverse drug resistance,K562/A02 cells were treated with PI3K inhibitor LY294002.It was found that LY249002 decreased ADR-induced γH2AX expression and increased the sensitivity of K562/A02 cells to ADR.Additionally,the single-cell gel electrophoresis assay and the Western blotting showed that LY249002 enhanced DSBs and decreased the expression of repair factor BRCAl.These results illustrate chemosensitivity can partly be measured by detecting γH2AX and drug resistance can be reversed by inhibiting γH2AX.
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This study examined the association of a common polymorphic allele (25G) of the low-density lipoprotein receptor-related proteinl (LRP1) gene with myocardial infarction (MI).The genotypes of LRP1 25CG (rs35282763) were determined in 347 MI patients and 347 age-and sex-frequency-matched controls from an unrelated Chinese Han population.Factor Ⅷ (FⅧ) levels were measured in the MI patients and controls by chromogenic assay and enzyme-linked immunosorbent assay (ELISA).The results showed that LRP1 25CG (rs35282763) genotype distribution did not differ significantly between patients (n=206 for 25CC,n=122 for 25CG) and controls (n=191 for 25CC,n=126 for 25CG;P＞0.05).The 25G allele was not associated with a reduced risk of MI (P ＞0.05).Further stratifications for age,sex,and other cardiovascular risk factors did not affect the negative findings.It was concluded that the presence of the G allele at the 25CG (rs35282763) polymorphism of the LRP1is not associated with a reduced risk of MI,and genotyping for LRP1 25CG (rs35282763) polymorphism is not useful in assessing the individual risk of MI.
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The binding function of EGF1 domain peptide with tissue factor(TF)and its ability of triggering coagulation were explored.The TF expression model in vitro was established by lipopolysaccharide induction.The affinity of EGFP-EGF1 and TF expressing cells was analyzed by fluorescence microscopy and flow cytometry(FCM).The affinity of EGFP-EGF1 and rat soluble TF was quantitated by surface plasmon resonance(SPR).The ability of EGFP-EGF1 in triggering coagulation was tested by prothrombin time assay.The FCM results showed recombinant factor Ⅶ(rFⅦ)could definitely depress the integration of EGFP-EGF1 with recombinant TF(rTF)(68.65%±3.86% vs 57.98%±4.71%,P<0.01).The SPR results indicated the association constant ka of EGFP-EGF1 proteins was higher than rFⅦ(8.29±1.39 vs 3.75±0.32,P<0.01).However,the EGFP-EGF1 protein lost the activity of triggering coagulation as compared with blood plasma of normal SD rats(56.8±3.2 s vs 17.8±3.4 s,P<0.01).It was concluded that the rat EGF1 peptide could specifically bind to TF without the ability of triggering coagulation.EGF1 peptide may be a good target head for delivering drugs to TF in anticoagulation therapy.