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Copyright of Food & Fermentation Industries is the property of Food & Fermentation Industries and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Journal of Food Safety & Quality is the property of Journal of Food Safety & Quality Editorial Department and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Objective: To establish a method for detecting the binding affinity of peptides to HLA-B*1301 by HLA molecular reconstitution after weak acid treatment, and its practicality was evaluated. Methods: C1R cells overexpressing HLA-B*1301 were treated with citrate buffer of different pH for different time. After neutralization of pH, cells were resuspended in culture medium containing β2m and brefeldin A. Cells were incubated at 37 °C with peptide (peptide group) or without peptide (blank control), after addition of anti-HLA antibody, the cells were detected by flow cytometry. Ratio of fluorescence intensity between peptide group and blank control group was used as an index to measure the level of HLA molecular remodeling, and then represented the interactions between peptide and HLA-B*1301 molecule. Results: HLA-polypeptide complex was denatalized and dissociated after being treated with pH3.0 buffer for 1 min, and only HLA heavy chain molecules were retained on the cell surface. The addition of peptides with binding force could significantly improve the level of HLA molecular remodeling, and the binding force of peptides with HLA-B*1301 could be evaluated by this method. Conclusion: HLA molecular reconstitution assay is a simple and reliable method to detect the binding of peptides to HLA-B*1301, which can also provide reference for the study of other low frequency HLA molecular antigen presentation patterns. [ABSTRACT FROM AUTHOR]

Objective: To establish a method for detecting the binding affinity of peptides to HLA-B*1301 by HLA molecular reconstitution after weak acid treatment, and its practicality was evaluated. Methods: C1R cells overexpressing HLA-B*1301 were treated with citrate buffer of different pH for different time. After neutralization of pH, cells were resuspended in culture medium containing β2m and brefeldin A. Cells were incubated at 37 °C with peptide (peptide group) or without peptide (blank control), after addition of anti-HLA antibody, the cells were detected by flow cytometry. Ratio of fluorescence intensity between peptide group and blank control group was used as an index to measure the level of HLA molecular remodeling, and then represented the interactions between peptide and HLA-B*1301 molecule. Results: HLA-polypeptide complex was denatalized and dissociated after being treated with pH3.0 buffer for 1 min, and only HLA heavy chain molecules were retained on the cell surface. The addition of peptides with binding force could significantly improve the level of HLA molecular remodeling, and the binding force of peptides with HLA-B*1301 could be evaluated by this method. Conclusion: HLA molecular reconstitution assay is a simple and reliable method to detect the binding of peptides to HLA-B*1301, which can also provide reference for the study of other low frequency HLA molecular antigen presentation patterns. [ABSTRACT FROM AUTHOR]

Copyright of Chinese Journal of Cancer Biotherapy is the property of Editorial Office of Chinese Journal of Cancer Biotherapy and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Chinese Journal of Cancer Biotherapy is the property of Editorial Office of Chinese Journal of Cancer Biotherapy and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Chinese Journal of Cancer Biotherapy is the property of Editorial Office of Chinese Journal of Cancer Biotherapy and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Chinese Journal of Cancer Biotherapy is the property of Editorial Office of Chinese Journal of Cancer Biotherapy and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Journal of Hydrobiology is the property of Editorial Department of Journal of Hydrobiology and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

【Objective】Chromosome ploidy identification is an important part of the evaluation of potato germplasm resources. The establishment of a high-throughput sample preparation method for the identification of potato chromosome ploidy based on flow cytometry would lay a foundation for the subsequent large-scale identification of potato ploidy.【Method】The 30 parthenogenetic offspring of potato were used to compare the effects of low-temperature steel-bullet beating, liquid nitrogen grinding and blade shredding to prepare nuclear suspensions. The ploidy of diploid potato IVP101 and tetraploid potato HTJ349-3 was identified.【Result】The suspension of cell nuclei prepared by the low-temperature steel-bullet beating method had obvious fluorescence signal, sufficient cell lysis and few impurities. Due to the simple and easy operation of the low-temperature steel-bullet beating method, the sample preparation time was greatly shortened, and the identification efficiency was 45-60 times higher than that of the liquid nitrogen grinding method, and 105-180 times higher than that of the blade shredding method. The chromosome ploidy of potato IVP101 and HTJ349-3 was accurately detected by this low-temperature steel-bullet beating method. 【Conclusion】The results by the low-temperature steel-bullet beating method are as accurate as that by the liquid nitrogen grinding method and the blade cutting method, which can achieve high-throughput identification of potato chromosome ploidy. [ABSTRACT FROM AUTHOR]

Objective: To develop multiplex cytokine detection reagents to analyze expression levels of cytokines, angiogene sis and bone remodeling proteins in relapse/refractory multiple myeloma (RRMM) . Methods: Multiplex bead-based immunoassay by flow cytometry was used to develop quantitative detection reagents of multiplex cytokines, which were applied to detect serum samples from 55 RRMM patients and 22 healthy controls. Expression levels of cytokines, angiogenesis, and bone remodeling proteins in pa tients, and their correlation with clinical characteristics were analyzed. Results: Detection reagents of 10-plex cytokine immunoassay were successfully developed in this study, with average sensitivity of 7.1 pg/ml, average recovery rate of 97.4%, average intra-assay CV of 4.8%, and average inter-assay CV of 9.0%. In addition, results of RRMM samples found that levels of IL-2, IL-17, DKK1, RANKL and OPG were positively correlated with the level of IgG monoclonal protein, and TIMP1 was positively correlated with levels of IgG and IgA monoclonal protein. Conclusion: In this study, ten kinds of cytokine detection reagents with high sensitivity and speci ficity are developed, and we found that IL-2, IL-17, DKK1, RANKL, OPG and TIMP1 have potential value in tracking disease pro gression in RRMM. The established development process of multiplex cytokine reagents has important reference significance for expanding the development and application of multiplex detection reagents for protein markers in the future. [ABSTRACT FROM AUTHOR]

Objective: To explore the molecular mechanism of dexmedetomidine (Dex) protecting ischemia/reperfusion (I/R) myocardium. Methods: Wild type mice were grouped into control (Control) group, sham operation (Sham) group, WT I/R group, WT Dex group, and Dectin-1 knock out mice were grouped into KO I/R group and KO Dex group in the in vivo study (n=6). TTC staining was used to determine the myocardial infarction area (%) of the above six groups of mice. HE staining and pathological analyze was used to determine the myocardial injury. Serum TNF-α, IL-6 and IL-10 levels in mice were detected by ELISA. Flow cytometry (FCM) was used to count and sort of infiltrating M2 macrophages and neutrophils in myocardium. qPCR assay was used to determine the Dectin-1 mRNA expression in the above sorted cells. Results: TTC results showed that there was no myocardial infarction in the mice of Control group and Sham group. Compared with the WT I/R group, the infarct volume was significantly lower in WT Dex group, KO I/R group and KO Dex group (P<0.05) . Compared with the KO I/R group, the infarct volume was reduced in KO Dex group (P< 0.05). The results of HE staining showed that the myocardial fibers of the WT I/R group of mice were disorderly arranged, with a large number of broken myocardial fibers, while the myocardial fibers of the WT Dex group, KO I/R group and KO Dex group of mice had a little breakage, the structural damage was not significant, and the myocardial arrangement was relatively neat. The degree of myocardi中国免疫学杂志 2024 年第 40 卷 al injury of mice in KO Dex group were less than that in KO I/R group mice. ELISA results showed that compared with Sham group, the serum TNF-α and IL-6 levels of the mice in WT I/R group were significantly increased, and the IL-10 level was significantly decreased. Compared with WT I/R group, serum TNF-α and IL-6 levels of the mice in WT Dex group and KO I/R group were significantly decreased, and IL-10 level was significantly increased. Compared with KO I/R group, the serum TNF-α and IL-6 levels of the mice in KO Dex group were significantly decreased, and the IL-10 level was significantly increased (P<0.05). FCM cell counting results showed that compared with Sham group, a large number of M2 macrophages and neutrophils were infiltrated in the myocardium of WT I/R group of mice (P<0.05) . Compared with WT I/R group, the M2 macrophages and neutrophils infiltrated in the myocardium were significantly decreased in WT Dex group, KO I/R group and KO Dex group of mice (P<0.05) . While there was no significant difference between the KO I/R group and the KO Dex group mice (P>0.05) . qPCR results showed that compared with Sham group, the expression level of Dectin-1 mRNA in the myocardial infiltrated M2 macrophages and neutrophils were significantly up-regulated in WT I/ R group of mice (P<0.05) . While compared with WT I/R group, the expression level of Dectin-1 mRNA in Dex group of mice was significantly lower (P<0.05) . Mice in KO I/R group and KO Dex group did not express Dectin-1. Conclusion: The protective mechanisms of Dex preconditioning on I/R injured myocardium involves reducing the infiltrating number of M2 macrophages and neutrophils in myocardium after I/R injury, which may be achieved by inhibiting the expression of Dectin-1. [ABSTRACT FROM AUTHOR]

Objective: To explore the mechanism of hepatitis B virus (HBV) inhibiting M1 macrophages to promote immune escape, and to provide targets and strategies for antiviral therapy. Methods: The human monocyte cell line THP-1 was induced into M1 macrophages with PMA+LPS+IFN-γ. Cell morphological changes and the expressions of CD68, CD86, HLA-DR and functional molecules IL-1β, IL-6, TNF-α in M1 macrophages were detected by flow cytometry and RT-qPCR to identify M1 macrophages. HBV stable replication cell line HepG2.2.15 were co-cultured with M1 macrophages, and the expression of HBV-DNA was detected by qPCR. The expression of CD68, CD86 and HLA-DR were detected by flow cytometry. The expressions of functional molecules TLR4, NLRP3, Caspase-1, pro-caspase-1, caspase-1 p20, IL-1β and IL-18 in M1 macrophages were determined by RT-qPCR and Western blot. Apoptosis rate was detected by flow cytometry, and the expression of apoptosis related protein cleaved-caspase-3 was determined by Western blot. Results: THP-1 was successfully induced to differentiate into M1 macrophages. M1 macrophages inhibited HBV replication (P<0.05). HBV inhibited the expressions of CD68, CD86 and HLA-DR in M1 macrophages（P<0.01). HBV inhibited the expressions of TLR4, NLRP3, Caspase-1, caspase-1 p20, IL-1β and IL-18 in M1 macrophages (P<0.01). HBV induced M1 macrophage apoptosis (P<0.05). Conclusion: HBV inhibits M1 macrophages and their functional molecules TLR4, NLRP3 and downstream factors, reduces the synthesis and secretion of inflammatory factors, induces apoptosis, and then promotes immune escape, resulting in the persistence and replication of HBV in the body. [ABSTRACT FROM AUTHOR]

Objective: To investigate the expression of group 2 innate lymphoid cells (ILC2s) and myeloid-derived suppressor cells (MDSCs), along with their associated cytokines IL-13 and inducible nitric oxide synthase (iNOS), in cervical cancer (CC), and to construct a nomogram prediction model for assessing the risk of CC based on these factors. Methods: Samples were collected from May 2022 to January 2024 at the First Affiliated Hospital of Xinjiang Medical University, including 40 cases of CC tissue and 100 cases of peripheral blood as the CC group. Concurrently, cervical tissues from 30 cases of uterine fibroids screened negative for CC and peripheral blood from 100 healthy individuals were selected as the control group. Multiple immunofluorescence technology (mIF) and immunohistochemical staining (IHC) were utilized to detect the infiltration of ILC2s and MDSCs in tissue samples from both groups, along with the expression levels of related cytokines IL-13 and iNOS. Flow cytometry (FCM) and ELISA were employed to assess the differences in ILC2s, MDSCs, IL-13, and iNOS expression in peripheral blood samples. Pearson correlation was used to assess their correlations. Univariate and multivariate logistic analyses were performed to determine whether ILC2s, MDSCs, IL-13, and iNOS are independent risk factors for CC. An immune prediction model was established using R software, and the model was evaluated using the area under the ROC curve (AUC value), Hosmer-Lemeshow test, calibration curve, clinical decision curve, and clinical impact curve.Results: The levels of ILC2, MDSC, IL-13 and iNOS were all significantly higher in the CC group compared to the control group (all P < 0.05). Furthermore, they were positively correlated (all P < 0.05). Univariate and multivariate logistic regression analyses indicated that ILC2, MDSC, IL-13, and iNOS are independent risk factors for the development of CC (all P < 0.05). Subsequently, a nomogram based on these risk factors was developed and verified to have practical clinical value. Conclusion: ILC2, MDSC, and their associated cytokines IL-13 and iNOS are highly expressed in CC tissues and peripheral blood. The prediction model incorporating these risk factors has predictive capabilities and clinical utility, providing a valuable and accessible tool for early diagnosis and treatment of cervical cancer. [ABSTRACT FROM AUTHOR]

Objective: To investigate the effects of mesenchymal to epithelial transition factors (MET) knockdown on the proliferation, migration, and sensitivity to 5-FU and cisplatin in human laryngeal squamous cell carcinoma (LSCC) Hep-2 cells. Methods: The expression of MET mRNA in LSCC tissues was analyzed using data from the Gene Expression Omnibus (GEO) and The Cancer and Tumor Gene Atlas (TCGA). Human normal bronchial epithelial cells (16HBE) and human LSCC cells (Hep-2, KBV200 and TU212) were routinely cultured, and the expression levels of MET in these cells were detected by qPCR and WB assay. The MET knockout plasmid (si-Met) and control plasmid (si-NC) were transfected into Hep-2 cells using Lipofectamine™ 3000, and the cells were divided into blank control group, si-NC group and si-Met group. The proliferation, migration, cell cycle distribution, and sensitivity to 5-FU and cisplatin of Hep-2 cells in each group were detected by MTT assay, flow cytometry and scratch assay, respectively. Results: Database analysis showed high expression of MET mRNA in LSCC tissue (P < 0.05). The mRNA and protein expression levels of MET in Hep2 cells, KBV200 cells and TU212 cells were significantly higher than those in 16HBE cells (all P < 0.01). After MET knockdown, the mRNA and protein levels of MET in Hep-2 cells were significantly reduced (P < 0.01 or P < 0.001), cell proliferation activity was significantly decreased (P < 0.000 1), the number of G0/G1 phase cells was significantly increased (P < 0.000 1), and the number of S phase cells was significantly reduced. Additionally, after MET knockdown, the inhibitory rates of cell proliferation by different concentrations of 5-FU or cisplatin were significantly enhanced, and the half maximal inhibitory concentration (IC50) was reduced (all P < 0.000 1). The scratch healing rate and migration ability were significantly reduced (all P < 0.05). Conclusion: MET is highly expressed in human LSCC tissues and cells. Knocking down MET can effectively inhibit the expression of MET in Hep-2 cells, suppress cell proliferation and migration ability, arrest the cell cycle in G1 phase, and enhance the sensitivity of Hep-2 cells to 5-FU and cisplatin. [ABSTRACT FROM AUTHOR]

Objective To isolate and culture neural crest cells (NCCs) from lung tissue of mice and to identify the characteristics of the cells in order to provide a new cell model for studying lung injury and injure repair. Methods The mT/mG dual-fluorescence reporter mice and Wnt1-Cre transgenic mice were hybridized, and mT/mG; Wnt1-Cre transgenic mice were screened to obtain enhanced green fluorescent protein (EGFP) permanently labeled NCCs. Cell suspension of mouse lung tissue was prepared by enzymolysis. EGFP+ cells (namely NCCs) were harvested by flow cytometry. Primary culture was performed with DMEM/F12 culture medium optimized in the laboratory, NCCs was characterized by immunofluorescence microscopy. Then NCCs differentiation was directed by mouse bone marrow mesenchymal stem cells osteogenic induction. Results The mT/mG of EGFP permanently labeled NCCs was successfully obtained by hybridization and high-purity NCCs were isolated from Wnt1-Cre transgenic mice lung tissue. They can be cultured in vitro and with spindle morphology which was,similar to fibroblast adherent proliferation. NCCs expressed the neural crest stem cell marker Sox10 and induced to differentiate into osteoblasts. Conclusions NCCs isolated and cultured from lung tissue of mT/mG;Wnt1-Cre transgenic mice show stable proliferation and have the characteristics of neural crest stem cells, which may function as a potential cell model for research on lung tissue injury and the mechanism of repair.

Copyright of Food & Fermentation Industries is the property of Food & Fermentation Industries and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Background and objective Isoliquiritigenin (ISL) is an important pharmacological constituent of Glycyrrhiza glabra, which possesses a range of physiological and pharmacological activities, as well as significant antitumor activity, and can be used as a potential drug for targeted cancer therapy. LINC01503 is an oncogene, which has been closely associated with the malignant biological processes of many cancers. The aim of this study was to investigate the effects of ISL on the proliferation, apoptosis, invasion and migration of lung squamous carcinoma cells by regulating LINC01503. Methods Plasma was collected from lung squamous carcinoma patients and healthy individuals treated at Tangshan People's Hospital from January 2021 to December 2022. The expression of LINC01503 in lung squamous carcinoma plasma, tissues and cells was detected by real-time quantitative fluorescence polymerase chain reaction (qRT-PCR). Lung squamous carcinoma cells were treated with different concentrations of ISL for 24 h, and LINC01503 expression was detected by qRT-PCR. The cells were treated in groups: si-NC group, si-LINC01503 group, DMSO (0.1% dimethyl sulfone) group, ISL group, pc DNA3.1(+)-NC group, pc DNA3.1(+)-LINC01503 group, ISL+pc DNA3.1(+)-NC group and ISL+pc DNA3.1(+)- LINC01503 groups. CCK-8 assay, clone formation assay, flow cytometry, Transwell assay and scratch assay were used to explore the effect of LINC01503 on the functional phenotype of lung squamous carcinoma cells. Results Fluorescence in situ hybridization results showed that the average fluorescence intensity of LINC01503 in tissue microarrays of lung squamous carcinoma patients was higher than that in paracancerous tissues (P<0.05). The expression of LINC01503 in the plasma of patients with lung squamous carcinoma was higher than that in the plasma of healthy individuals (P<0.05). Knockdown of LINC01503 inhibited the proliferation, invasion and migration of lung squamous carcinoma cells and promoted apoptosis (P<0.05). ISL inhibited the proliferation, invasion, migration and promoted apoptosis of lung squamous carcinoma cells (P<0.05). Overexpression of LINC01503 followed by intervention with ISL reversed the promotional effect of overexpression of LINC01503 on the proliferation, invasion and migration of lung squamous carcinoma cells as well as the inhibitory effect on apoptosis (P<0.05). Conclusion LINC01503 was highly expressed in lung squamous carcinoma, and LINC01503 could promote the proliferation, invasion and migration of lung squamous carcinoma cells and inhibit the apoptosis, ISL could inhibit the proliferation, invasion and migration of lung squamous carcinoma cells and promote apoptosis of lung squamous carcinoma cells by regulating the expression of LINC01503. [ABSTRACT FROM AUTHOR]

Copyright of Chinese Journal of Cancer Biotherapy is the property of Editorial Office of Chinese Journal of Cancer Biotherapy and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Chinese Journal of Cancer Biotherapy is the property of Editorial Office of Chinese Journal of Cancer Biotherapy and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Chinese Journal of Cancer Biotherapy is the property of Editorial Office of Chinese Journal of Cancer Biotherapy and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Objective This study is assigned to explore the expression of CD85d in acute myeloid leukemia and its clinical significance in the diagnosis of monocyte associated acute myeloid leukemia. Methods We used flow cytometry by CD45/SSC to analyze the expression of CD85d, CD14, CD64 in 46 monocyte associated acute myeloid leukemia (M-AML) and 60 non monocyte associated acute myeloid leukemia (NM-AML) patients admitted to the Hematology Department of Hainan General Hospital from March 2022 to December 2023. Results Both normal granulocytes and normal monocytes express CD85d, and monocytes express fluorescence with stronger intensity than granulocytes. CD85d is not expressed in normal lymphocytes. The positive expression rate of CD85d in M-AML was 80.4% (37/46), which was significantly higher than that in NM-AML 3.3% (2/60), and the difference was statistically significant (P<0.01). The sensitivity of diagnosing M-AML was ranked from high to low as CD64 (87.0%)>CD85d (80.4%)>CD14 (34.8%). The specificity of diagnosing M-AML was ranked from high to low as with CD14 (100%)>CD85d (96.7%)>CD64 (56.7%). The sensitivity and specificity of CD85d combined with CD64 in diagnosing M-AML were 89.1% and 56.7%. There was no statistically significant difference between the detection rate of abnormal karyotypes (53.8%), positive rate of the fusion genes(26.9%), the CR rate (78.3%) in CD85d+ AML and that in CD85d- AML (66.0%, 50.0%, 70.0%) (P>0.05). Conclusion CD85d can be used as a new high sensitivity and specific surface marker for immature monocytes, which is helpful to improve the detection rate of M-AML and differential diagnosis with NM-AML subtypes. [ABSTRACT FROM AUTHOR]

The maintenance and variation of ploidy in plants are influenced by environmental factor, and the tissue culture regeneration can often cause changes of chromosome structures and ploidy levels. In order to explore the ploidy variation of Camellia germplasm under tissue culture conditions, the callus induction system of Camellia germplasm was used to analyze the ploidy changes by flow cytometry, and the stability and variation of the ploidy under tissue culture generation were analyzed in combination with colchicine treatment. The results were as follows: (1)Six of ten of Camellia germplasms were diploid, two of them were tetraploid, one hexaploid and one decaploid, and the germplasm materials largely maintained stable ploidy levels during the tissue regeneration processes. (2)The optimum conditions for colchicine treatment were obtained, that is, the callus was cultured in treatment medium containing colchicine at 20 mg·L-1 for 10 d, followed by regeneration in tissue-differentiation culture. (3)The ploidy analyses were carried out on the treated tissues in different differentiation states, and the results showed that most of the independent tissues(including 56 callus and shoots)were found to be aneuploids, 38 tissues had ploidy between 1.5 to 2.5, and 11 tissues produced less than 1.5. This study further explores the ploidy relationship between different Camellia germplasms, and provides a reference for an in-depth study of ploidy regulation and polyploid induction of Camellia. [ABSTRACT FROM AUTHOR]

Objective To investigate the differences of characteristic of bone marrow plasma cells in patients of different plasma cell dyscrasias according to International Myeloma Working Group (IMWG) criteria. Methods We analyzed the serological and bone marrow flow cytometry results of patients with plasma cell dyscrasias diagnosed and treated in Department of Hematology, Zhongshan Hospital (Xiamen Branch), Fudan University from Jun 12, 2019 to Sep 5, 2023 retrospectively. Results A total of 102 patients, 63 males and 39 females, aged 22 to 85 years, were included, including 46 patients with monoclonal gammaglobulinemia of unknown significance, 39 patients with multiple myeloma, 5 patients with smoldering multiple myeloma and 12 patients with light-chain amyloidosis. All patients had M proteinemia, including 58 patients with IgG type and 44 patients with non-IgG type. Plasma cells were detected in the bone marrow of all patients. Clonal plasma cells were detected in the bone marrow of 79 patients. Normal plasma cells were detected in the bone marrow of 63 patients. Both clonal and normal plasma cells were detected in the bone marrow of 40 patients. Clonal plasma cells from bone marrow of 52 patients expressed CD56 and 12 patients expressed CD117. There were no significant differences in gender, age among different disease groups. There were statistical differences in M protein type, the concentration of M protein, serum involved/uninvolved free light chain ratio, the proportion of plasma cells in bone marrow nucleated cells, the proportion of clonal plasma cells in bone marrow nucleated cells, the proportion of clonal plasma cells in all bone marrow plasma cells, and the proportion of normal plasma cells in all bone marrow plasma cells among different disease groups (P<0.05). There was statistical difference in the expression of CD56 in clonal plasma cells among different disease groups (P=0.009), but no statistical difference in the expression of CD117. Conclusion The proportion of clonal plasma cells to all nucleated cells, the proportion of clonal plasma cells to all plasma cells, and the proportion of CD56 expression in abnormal plasma cells in the bone marrow of patients with light amyloidosis were similar to those of monoclonal gammopathy of undetermined significance, but significantly different from those of patients with multiple myeloma. [ABSTRACT FROM AUTHOR]

Objective: To investigate whether galangin (Gal) affects the proliferation, migration, invasion and apoptosis of osteosarcoma MG63 cells by regulating the cGAS/STING signaling pathway. Methods: Human osteosarcoma MG63 cells were cultured in vitro and treated with Gal at concentrations of 0, 5, 15, 25, 50, 100 and 200 μmol/L for 48 hours, and the effect of Gal on cell viability was detected by CCK-8 method. MG63 cells were divided into the control group (untreated cells), the Gal low concentration group (Gal-L group, treated with 50 μmol/L Gal), the Gal high concentration group (Gal-H group, treated with 100 μmol/L Gal), and the Gal-H+STING inhibitor group (Gal-H+H-151 group, treated with 100 μmol/L Gal+8 μmol/L H-151). EdU staining method, scratch healing test, Transwell chamber method and flow cytometry were used to detect cell proliferation, migration, invasion and apoptosis of cells in each group. Western blot was applied to detect the expression levels of cGAS and STING proteins in cells of each group. Results: Compared with the control group, the proliferation activity, migration, and invasion abilities of the cells in the Gal-L and Gal-H groups were significantly decreased (all P<0.05); the apoptosis rate and the expression levels of cGAS and STING proteins were significantly increased (all P<0.05). Compared with the Gal-H group, the proliferation activity, migration, and invasion abilities of the cells in the Gal-H+H-151 group were significantly increased (all P<0.05); the apoptosis rate and the expression levels of cGAS and STING proteins were significantly decreased (all P<0.05). Conclusion: Gal may inhibit the proliferation, migration and invasion and promote the apoptosis of osteosarcoma cells by activating the cGAS/STING signaling pathway. [ABSTRACT FROM AUTHOR]

Objective: To explore the effect of Dickkopf-1 (DKK1) expression on CD4+ T cell polarization in malignant tumors and its potential value as a target for cancer immunotherapy. Methods: Bioinformatics algorithm was used to analyze the expression of DKK1 in multiple types of tumor tissues and adjacent tissues, and the correlation between the expression of DKK1 and the prognosis of cancer patients and immune infiltration of tumor microenvironment. The effect of DKK1 protein on the phenotype of CD4+ T cells was analyzed by flow cytometry in vitro. A melanoma B16F10 cell mouse subcutaneous transplantation tumor model was established to observe the effects of blocking DKK1 on the growth of mouse transplantation tumor and immune cell infiltration and phenotype in transplantation tumor tissues. Results: The mRNA expression levels of DKK1 in many kinds of tumor tissues were significantly higher than those in adjacent tissues. High expression of DKK1 was related to the poor prognosis of most tumor patients. In most types of tumors DKK1 played important negative regulatory role in CD4+ T cell anti-tumor immune response (P<0.05 or P<0.01). The results of flow cytometry in vitro showed that DKK1 protein stimulation significantly decreased the expression levels of T-bet, IFN-γ and CD107a in CD4+ T cells (all P<0.01). In the mouse subcutaneous melanoma model, it was found that blocking DKK1 could significantly inhibit the growth of transplanted tumors (P<0.01) and effectively improve the anti-tumor immune responses including the proportion of Th1 cells (T-bet+CD4+) increasing significantly (P<0.001), the proportion of effector CD8+ T cells (CD44+CD62L-) increasing significantly (P<0.01), and the proportion of Th2 cells (GATA3+CD4+) and Treg cells decreasing significantly (both P<0.01). Conclusion: Blocking DKK1 can effectively promote the phenotypic polarization of CD4+ T cells to Th1. DKK1 has potential value as a target for tumor immunotherapy. [ABSTRACT FROM AUTHOR]

Objective: To explore the effects of UCP2 inhibitor genipin (GEN) on the proliferation and mitochondrial function of human hypopharyngeal carcinoma FaDu cells. Methods: FaDu cells were treated with different concentrations of GEN for 24 hours and divided into the GEN 0 μmol/L (control) group, the 50 μmol/L group, the 100 μmol/L group, the 200 μmol/L group and the 400 μmol/L group. The CCK-8 method was employed to assess cell proliferation, and the DCFH-DA probe and JC-1 flow cytometry were utilized to measure the impact of GEN on reactive oxygen species (ROS) levels and mitochondrial membrane potential in FaDu cells. Laser confocal microscopy was utilized to observe the effect of GEN on the mitochondrial membrane permeability transition pore (MPTP) in FaDu cells. Spectrophotometry was employed to measure lactate levels in cells, and Western blot analysis was conducted to monitor changes in UCP2 protein expression. Results: In comparison with the control group, GEN significantly inhibited the proliferation activity of FaDu cells (P<0.05, P<0.01), reduced the expression of UCP2 protein in cells (P<0.05), decreased the mitochondrial membrane potential (P<0.05, P<0.01) and lactate content (P<0.001), altered mitochondrial membrane permeability, and increased the levels of ROS in cells (P<0.05, P<0.01). Conclusion: GEN modulates the expression of UCP2 in cells, consequently altering their redox potential and mitochondrial function, thus inhibiting the viability and inducing apoptosis in human nasopharyngeal carcinoma FaDu cells. [ABSTRACT FROM AUTHOR]

Objective: To investigate the effects of isoliensinine (Iso) on the proliferation, apoptosis and autophagy of colon cancer SW480 cells through PI3K/Akt/mTOR signaling pathway. Methods: Colon cancer SW480 cells were treated with 10, 20 and 40 μmol/L Iso, and the effects of Iso on the cell proliferation capacity, apoptosis and expressions of autophagy related proteins LC3 I, LC3II and p62 were detected by CCK-8, flow cytometry and Western blot, respectively. Then, SW480 cells were treated respectively with 20 μmol/L Iso and 25 μmol/L PI3K activator 740 Y-P, and the cells were divided into the control group, the 740 Y-P group, the Iso group and the Iso+740 Y-P group. The effects of 740 Y-P on the apoptosis and the expressions of LC3 I, LC3 II, p62, PI3K, p-PI3K, mTOR and p-mTOR proteins in each group were detected by flow cytometry and WB. Results: After treatments with 10, 20 and 40 μmol/L Iso, the proliferation capacity of SW480 cells was decreased significantly (all P<0.05); the apoptosis rate was increased significantly (all P<0.05); the expressions of LC3 II/LC3 I were up-regulated significantly (all P<0.05), and the expression of p26 protein was down-regulated significantly (all P<0.05). After treatments with Iso and 740 Y-P, compared with the control group, the apoptosis rate and LC3 II/LC3 I expression of the 740 Y-P group were decreased significantly (both P<0.05), while the expressions of p26, p-PI3K/PI3K and p-mTOR/mTOR were increased significantly (all P<0.05).The apoptosis rate and LC3 II/LC3 expression in the Iso group were increased (both P<0.05) and the expressions of p26, p-PI3K/PI3K and p-mTOR/mTOR were decreased (all P<0.05). Compared with the 740 Y-P group, the apoptosis rate and LC3 II/LC3 I expression were increased in the Iso+740 Y-P group (P<0.05), while the expressions of p26, p-PI3K/PI3K and p-mTOR/mTOR were decreased (all P<0.05). Compared with the Iso group, the apoptosis rate and LC3 II/LC3 I expression were decreased (both P<0.05), and the expressions of p26, p-PI3K/PI3K, and p-mTOR/mTOR were increased significantly (all P<0.05) in the Iso+740 Y-P group. Conclusion: Iso inhibits the proliferation and induces the apoptosis and autophagy of SW480 cells by inhibiting PI3K/Akt/mTOR signaling pathway. [ABSTRACT FROM AUTHOR]

Objective: To design and construct CD19-targeting CAR-T cells expressing PD-1 shRNA and validate their anti-tumor function in vitro. Methods: The authors designed and constructed CD19 CAR molecule gene expressing PD-1 shRNA, and packaged them into retroviral vector using packaging cells. The viral vector copy number was detected by qPCR, and then human primary T cells were transduced to obtain CAR-T cells, which were divided into three groups: RNAU6-CD19 CAR-T, PD-1 shRNA1-CD19 CAR-T, and PD-1 shRNA2-CD19 CAR-T cells. qPCR was applied to detect the expression levels of PD-1 mRNA in three groups of CAR-T cells. Flow cytometry was used to detect the expression level of PD-1 on CAR-T cells in three groups. The luciferase reporter gene method and flow cytometry were used to detect the killing function of CAR-T cells against target cells (human lymphoma daudi cells) at different efficacy to target ratios. Results: Three groups of CAR molecules, namely RNAU6-CD19 CAR, PD-1 shRNA1-CD19 CAR and PD-1 shRNA2-CD19 CAR, were successfully packaged into retroviral vector, in which all retroviral vector copy numbers were higher than 1x107 copies/mL. CAR-T cells were obtained by transducing human primary T cells. The CAR-T transduction efficiencies of RNAU6-CD19 CAR-T, PD-1 shRNA1- CD19 CAR-T and PD-1 shRNA2-CD19 CAR-T cells were 43.1%, 55.1%, and 41.7% respectively. Compared with RNAU6-CD19 CAR-T cells, PD-1 shRNA1-CD19 CAR-T and PD-1 shRNA2-CD19 CAR-T cells showed a significant decrease in the expression level of PD-1 mRNA (all P<0.01), lower expression level of PD-1 on cell surface s (all P<0.01), and higher killing efficiency against CD19 positive target cells (daudi cells) in vitro (P<0.05 or P<0.01). Conclusion: Successfull construction of CD19-targeting CAR-T cells expressing PD-1 shRNA can improve the killing efficiency against CD19 positive target cells, reduce the expressions of PD-1 mRNA and its translation product PD-1, and slow down the depletion of CAR-T cells. [ABSTRACT FROM AUTHOR]

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Objective : To observe the effect of Yupingfeng powder on ZBP1-mediated PANoptosis of alveolar macrophages so as to elucidate its molecular mechanism of inhibiting airway inflammation in asthma. Methods : Lipopolysaccharide was used to induce alveolar macrophages (MH-S) to construct an asthma cell model, and different concentration of Yupingfeng powder medicated Serum were given for intervention. CCK8 method was used to detect the activity of NH-S cells, ELISA method was used to detect the content of LDH, IL-1β and IL-18, flow cytometry was used to detect cell death, Western blot method was used to detect the expression of PANoptosis-related proteins and ZBP1 protein, and RT-qPCR method was used to detect the expression of ZBP1 gene. Results: Compared with the control group, the LDH release of alveolar macrophages, the content of IL-1β and IL-18 and the proportion of dead cells in the model group were significantly increased. The expressions of pyroptosis proteins (Cleaved caspase-1 and GSDMD-NT), necroptosis proteins (p-RIPK3 and p-MLKL), and apoptosis protein (Cleaved caspase-3) were significantly up-regulated (P<0.01). Compared with the model group, the expression of LDH release, the content of IL-1β and IL-18, and the proportion of dead cells in alveolar macrophages in the Yupingfeng powder group were significantly reduced, and the expression of Cleaved caspase-1, GSDMD-NT, p-RIPK3, p-MLKL, and Cleaved caspase-3 were significantly down-regulated (P<0.01). In addition, the expression of ZBP1 protein and ZBP1 mRNA in alveolar macrophages in the Yupingfeng powder group was significantly reduced compared with the model group, and the differences were statistically significant (P< 0.01). Conclusion: Yupingfeng powder ameliorates airway inflammation by inhibiting the expression of ZBP1 in alveolar macrophages, thereby suppressing the level of PANoptosis. [ABSTRACT FROM AUTHOR]

Copyright of Chinese Journal of Cancer Biotherapy is the property of Editorial Office of Chinese Journal of Cancer Biotherapy and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Chinese Journal of Cancer Biotherapy is the property of Editorial Office of Chinese Journal of Cancer Biotherapy and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Chinese Journal of Cancer Biotherapy is the property of Editorial Office of Chinese Journal of Cancer Biotherapy and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Objective To examine the expression levels of Toll-like receptor 5 (TLR5) on peripheral blood monocytes in patients with rheumatoid arthritis (RA), and to explore its clinical significance. Methods A total of 140 RA patients, along with 70 healthy controls (normal control group) received in Suining Central Hospital from January 2017 to August 2022 were selected. The RA patients were divided into high disease activity group (n=70) and moderate-to-low disease activity group (n=70) based on the disease activity score 28-erythrocyte sedimentation rate (DAS28-ESR). Flow cytometry was used to determine the positive rate of TLR5 on peripheral blood monocytes among three groups. The predictive value of TLR5 expression on monocytes surface for RA was analyzed using receiver operating characteristic (ROC) curves. Results The positive rates of TLR5 on the surface of monocytes in normal control group, moderate-to-low disease activity group and high disease activity group were (46.99±5.79)%, (50.80±5.96)% and (58.86±6.20)%, respectively, and there was a statistically significant difference among the three groups (F=71.550, P＜0.01). The area under the ROC curve for TLR5 in diagnosing RA was 0.79, with a cut-off value of 49.50%, sensitivity of 72.86%, and specificity of 73.57%. Conclusion The TLR5 expressed on peripheral blood monocytes is involved in the pathogenesis and maintenance of disease activity and has predictive value for RA.

Objective: To investigate the effects of wild-type vesicular stomatitis virus strain (VSV-IN) on immunomodulation and tumour metastasis in mouse triple negative breast cancer (TNBC) 4T1 cell transplantation model mice. Methods: After VSV-IN infected 4T1 cells with MOI=1, MOI=10 and MOI=100 for 12, 24 and 48 h, 4T1 cell mortality was detected by CCK-8 method, migration ability by scratch healing assay, and the expressions of E-cadherin, MMP-2 and MMP-9 mRNA by qPCR. The fat pads of female BALB/c mice were inoculated with 0.1 mL of 4T1 cells with a cell density of 1x106 cells/mL to construct a 4T1 cell-loaded mouse model. When the tumour volume reached 200 mm³, the mice were injected intratumorally with PBS, taxol (TAX) (15 mg/kg), and VSV (1x106 pfu) once a week, respectively. After 4 administrations, mice were executed, stripped of intact grafted tumour tissues, and tumour volume and mass were measured. Histopathological sections of the lungs were stained with H-E, and tumour metastatic nodules of the lungs were observed. The proportion of T-cell subpopulations in the spleen was detected by flow cytometry. The levels of serum IL-6 and TNF-α were detected by ELISA. The expression levels of migration-related proteins in mammary gland tumours were analysed by using the online analysis of GEPIA. The expressions of MMP-2, MMP-9 and E-cadherin in mouse tumours were detected by immunohistochemistry, and the affinity of G and M proteins of VSV-IN and ERK2 and E-cadherin was predicted by protein-protein docking technology. Results: The mortality rate of 4T1 cells after 48 hours of VSV treatment at MOI=10 and 100 were significantly higher than that of the control group (P<0.01). Compared with that of the control group, the cell migration rate of the VSV-IN group (MOI=10) was significantly lower (P<0.01), and the relative expression of MMP-9 mRNA was significantly lower (P<0.05). Compared with those in the mice of the control group, transplanted tumours in the mice of the VSV-IN group grew more slowly, and their endpoint volume was significantly reduced (P<0.05). The number of lung metastatic nodules in the mice of the VSV group was significantly less than that of the control group ([12.86±1.86] vs [24±3.67], P<0.01). The proportions of splenic CD4+ T and CD8+ T cells in the VSV group were significantly higher (both P<0.05). The serum TNF-α and IL-6 levels were significantly higher (both P< 0.01). GEPIA tool analysis revealed that the expression levels of E-cadherin and MMP-9 were higher in breast cancer tissues than in paracancerous tissues (P<0.05). The expression of MMP-9 in the tumour cells of the mice in the VSV-IN group was significantly lower than that in the control group (P<0.05). The binding free energies of G and M proteins of VSV-IN to ERK2 were -11.7 kcal/mol and -6.4 kcal/mol, respectively. Conclusion: Wild-type VSV-IN inhibits the growth and metastasis of transplanted tumours in 4T1 tumor-bearing mice, which may be related to its promotion of anti-tumour immunity and modulation of the expression of migration-related proteins. [ABSTRACT FROM AUTHOR]

Objective: To engineer a BCMA mutant resistant to γ-secretase cleavage in order to stabilize wild-type BCMA expression after γ-secretase cleavage and to generate target cells for measuring BCMA CAR-T cell cytotoxicity. Methods: Our study aimed to engineer a mutant BCMA protein (BCMA-CD8α TM) that could resist γ-secretase cleavage by replacing the transmembrane domain of the wild-type BCMA protein with the human CD8α sequence. Four different types of cells in which this mutant gene was expressed excessively were engineered, including U266 (U266BCMA Mut), K562 (K562BCMA Mut), SKOV3 (SKOV3BCMA Mut), and CHO (CHOBCMA Mut) cells. BCMA CAR Jurkat cells, loaded with the NFAT-EGFP reporter gene (BCMA-CAR-Jurkat-Reporter), were engineered and cocultured with U266BCMA Mut cells. The expression level of EGFP was detected by FCM in order to indicate the activation level of NFAT. The cytotoxicity of BCMA CAR-T cells against Luciferase-labeled K562BCMA Mut cells was detected by the luciferase assay. Additionally, real-time cell analysis (RTCA) technique was employed to detect the cytotoxicity of BCMA CAR-T cells against SKOV3BCMA Mut and CHOBCMA Mut cells. Results: Application of γ-secretase inhibitor LY411575 to inhibit γ-secretase activity significantly enhanced the expression level of BCMA on the surface of wild-type U266 cells, and the average fluorescence intensity was increased by more than 10 times. However, the expression level of BCMA gradually decreased after removal of inhibitors (P<0.01). BCMA-CD8α TM mutant could resist the cleavage of γ-secretase and expressed stably on the surface of U266 cells (P>0.05). U266 cells and U266 cells overexpressing BCMA-CD8α TM were co-incubated with BCMA-CAR-Jurkat-Reporter cells, both of which could activate the Reporter system and enhance the expression of EGFP, but the effect was more significant in U266 cells overexpressing BCMA-CD8α TM (P<0.01). BCMA-CD8α TM mutants were successfully overexpressed in 3 BCMA-negative target cells, namely K562, SKOV3 and CHO cells, and the expression level of the mutant was only slightly increased under LY411575 treatment. Luciferase assay results showed that under different target-effect ratios BCMA CAR-T cells could all be specific and efficient in killing K562 cells overexpressing BCMA-CD8α TM. RTCA results showed that under different target ratios BCMA CAR-T cells could all effectively recognize and kill SKOV3 and CHO cells overexpressing BCMA-CD8α TM, but Mock-T cells with the same target ratio had no such effect. Conclusion: The BCMA-CD8α TM mutant engineered in this study can resist γ-secretase cleavage and exhibits stable surface expression on various target cells, thus offering various methods for evaluating the efficacy and specificity of BCMA CAR-T cell cytotoxicity in vitro. [ABSTRACT FROM AUTHOR]

Objective: To study, through bioinformatics analysis and cellular biology experiments, the effects of keratin 6A (KRT6A) on the diagnosis, prognosis and immune microenvironment of pancreatic ductal adenocarcinoma (PDAC) and biological behaviors of PDAC PANC1 cells, such as proliferation and apoptosis. Methods: The GEPIA platform was used to integrate the data from the TCGA (The Cancer Genome Atlas) and the GTEx (Genotype-Tissue) to analyze the KTRT6A expression in PDCA tissues. CIBERSORT software was then used to analyze the correlation between KRT6A expression and immune cell infiltration in PDCA tissues, and GSEA analysis was used to study the signaling pathway related to KRT6A gene expression. Immunohistochemical analysis was performed on cancer and adjacent tissue samples from 60 PDAC patients preserved in the Department of Pathology of Changhai Hospital to verify the expression of KRT6A in tumor tissues. The expression of KRT6A gene in PANC1 cells was inhibited by interfering with siRNA. The effects of KRT6A on the proliferation and apoptosis of PDAC cells were detected by CCK-8 assay and flow cytometry. Results: Data analysis of the TCGA and the GTEx found that KRT6A was highly expressed in human PDAC tissues and was significantly correlated with poor survival period of patients (P=0.015). CIBERSORT software and GSEA analysis showed that the infiltration of M2-type macrophages in PDCA tissues with high KRT6A expression was increased (P=0.034), and there was a significant correlation between high KRT6A expression and the up-regulation of Wnt pathway (NES: 1.7359272, P<0.05), pentose phosphate pathway (PPP) (NES: 1.5613053, P<0.05) and other signaling pathways (P<0.05 or P<0.01). Immunohistochemical results further verified the high expression of KRT6A in PDCA tissues (P<0.001). Proliferation and apoptosis experiments showed that interfering with KRT6A gene could significantly inhibit the proliferation (P<0.05) and apoptosis (P<0.001) of PANC1 cells. Conclusion: KRT6A is highly expressed in human PDAC tissues, and knocking down its expression can inhibit the proliferation and apoptosis of PANC1 cells, and has the potential to be a new target for PDAC diagnosis and prognosis. [ABSTRACT FROM AUTHOR]

Objective: To explore the effect of midchain fatty acid decanoic acid on CD8+ T cell activation and its effect and mechanism in CD8+ T cell-mediated anti-tumor immune response. Methods: Subcutaneous melanoma B16F10 cells tumor-bearing C57BL/6 mouse models were established and randomly divided into the decanoic acid group (10 mg/kg decanoic acid by gavage) and the control group (equal amount of solvent by gavage). The effect of decanoic acid on the growth of mouse tumors and their survival rate were measured. The activation of tumor-infiltrated CD8+ T cells in the tumor microenvironment was detected by flow cytometry. The α-CD8 mAb was used to deplete CD8+ T cells in B16F10 tumor-bearing mice, and the effect on the tumor volume was observed. Mouse primary CD8+ T cells were treated with decanoic acid, and T cell receptor (TCR) activation, effector cytokine production as well as proliferation and metabolism levels were detected by WB, ELISA, qPCR, and flow cytometry. In B16F10 tumor-bearing mouse model, the effects of administration of α-PD-1 mAb combined with decanoic acid on the growth of mouse tumors and mouse survival rate were observed. Results: In the mouse melanoma model, compared with those in the control group the volume of mouse transplanted tumors significantly reduced and mouse survival rate significantly increased in the decanoic acid group. (both P<0.05). The expression levels of IFN-γ and TNF-α in tumor-infiltrating CD8+ T cells were significantly higher in the decanoic acid group than that in the control group (P<0.01). The killing ability of OT-I T cells against B16F10-OVA cells was significantly elevated after treatment with decanoic acid (P<0.01). The suppressive effect of decanoic acid on transplanted tumors was significantly reduced after CD8+ T cells were depleted with α-CD8 mAb in the melanoma mouse model (P<0.000 1). Mouse-derived primary CD8+ T cells treated with decanoic acid showed significantly higher levels of TCR activation, increased production of cytokines IL-2 and IFN-γ, and significantly up-regulated the mitochondrial metabolic level (all P<0.05). In the melanoma mouse model, decanoic acid in combination with α-PD-1 mAb significantly inhibited tumor growth and increased the survival rate (both P<0.05). Conclusion: Decanoic acid can enhance the anti-tumor immune responses by promoting CD8+ T cell activation. [ABSTRACT FROM AUTHOR]

Objective: To investigate the effects of berberine (BBR) combined with XAV939 on the migration and apoptosis of osteosarcoma MG-63 cells and its possible mechanisms. Methods: MG-63 cells were cultured, and 20~120 µmol/L of BBR and 5~60 µmol/L of XAV939 were added, respectively. The cytotoxicity of BBR and XAV939 was determined by CCK-8 assay, and the combined index of the two was clarified using Chou-Talalay analysis to determine the intervention dose for the subsequent experiments. MG-63 cells were randomly divided into the blank control (NC) group, the BBR group, the XAV939 group, and the BBR+XAV939 combined group. The effects of BBR and XAV939 treatment alone or in combination on the migratory ability of MG-63 cells were detected by the scratch healing assay and the Transwell assay. Hoechst 33258 staining, JC-1 staining and Annexin V-FITC/PI double staining flow cytometry were used to detect the effects on cell mitochondrial membrane potential and apoptosis. Immunofluorescence was used to datect the expression of BAX protein, WB assay was used to detect the effects on the expression of MMP-2 and BAX in the cells, and qPCR was used to detect the effects on the expression of the MMP-2 gene. Results: BBR and XAV939 inhibited the proliferation of MG-63 cells in a dose-dependent manner, and 30 µmol/L BBR, 7.5 µmol/L XAV939 were selected for subsequent experiments. Compared with the NC group, 24 h after the cells were treated with BBR (30 µmol/L), XAV939 (7.5 µmol/L) and BBR+ XAV939 combination, the migration ability of MG-63 cells was significantly reduced; apoptotic cells were significantly increased (all P <0.05); the mitochondrial membrane potential was decreased (P<0.01); the gene expression of MMP-2, and MMP-2, BAX protein levels were all reduced (all P<0.05). In addition, the effect of the combination group was significantly stronger than that of the monotherapy group (P<0.05 or P<0.01). Conclusion: BBR and XAV939 alone or in combination can inhibit the migration and promote the apoptosis of MG-63 cells. The mechanism may be related to the decreased expression of migration-related protein MMP-2 and increased level of apoptosis-related protein BAX. [ABSTRACT FROM AUTHOR]

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Extracellular vesicles (EVs) were isolated from the culture supernatant of lung adenocarcinoma A549 cells. The size of EVs was characterized as approximately 100 nm via DLS and TEM analysis, with the EV-specific marker TSG101 detected in the samples. The extracted EV DNA (evDNA) showed a predominantly ~ 12 kb size distribution as assessed by agarose gel electrophoresis. Based on sequencing data, specific evDNA was selected for PCR detection. DNase I or PS nuclease treatment of the EVs confirmed that the evDNA existed on the outside of the vesicle in a linear manner, and poly-dA tails were added to evDNA by TdT enzyme treatment, allowing for their capture by oligo-dT beads and presenting further evidence of their linear surface distribution. Additionally, a TetO-DNA/TetR-GFP visualization system was established to confirm the presence of TetO-DNA in the EVs, which could be visualized by TetR-GFP proteins under fluorescent microscopy. Enrichment of EVs carrying the TetO-DNA was confirmed via flow cytometry and PCR experiments using TetR-GFP protein. These findings collectively suggest that evDNA is distributed linearly on the surface of EVs, providing a basis for further research into their biological functions. [ABSTRACT FROM AUTHOR]

Objective To investigate the expression and clinical significance of lymphocyte-derived microvesicles (LMP) in patients with multiple myeloma (MM). Methods A total of 65 newly diagnosed MM patients were used as the initial diagnosis group, and 30 health examination volunteers were selected as the control group. After 4 courses of chemotherapy, 8 patients in the initial diagnosis group died within 3 months, and the remaining 57 patients were included in the post chemotherapy group. Flow cytometry was used to detect the expression of LMP in peripheral blood of three groups, as well as the immune phenotype expression of peripheral blood lymphocyte subsets and bone marrow MM cells in the initial diagnosis group. Values of LMP between different therapeutic groups after chemotherapy were compared. The receiver operating characteristic (ROC) curve was used to determine the cutoff value for predicting death for each LMP. For the high (H) group with LMP ≥ cut-off value and the low (L) group with LMP＜cut-off value, Kaplan Meier survival analysis was performed. Variables with a P value＜0.05 in Kaplan Meier analysis were included into Cox regression analysis to analyze influencing factors of patient death. The differences in lymphocyte subpopulations and myeloma cell immunophenotypes were compared between different LMP groups. Results The proportions of LMP, CD3+ LMP and CD3+ CD8+ LMP were lower in the initial diagnosis group than those in the control group, while the proportions of NKLMP and CD4+ /CD8+ LMP were higher than those in the control group (P＜0.05). The proportion of CD3+ CD8+ LMP was higher in the post chemotherapy group than that in the initial diagnosis group, while the proportion of CD3+ CD4+ LMP and the ratio of CD4+ /CD8+ LMP were lower than those in the initial diagnosis group (P＜0.05). The proportion of CD3+ LMP and ratio of CD3+ CD8+ LMP were higher in the CR+VGPR group than those in the PR+MR+PD group (P＜0.01), while the proportion of NKLMP and ratio of CD4+ /CD8+ LMP were lower than those in the PR+MR+PD group (P＜0.05). Kaplan Meier analysis showed that the median survival time (OS) was shorter in the LLMP group than that of the HLMP group (P＜0.01). The median OS was shorter in the LNKTLMP group than that in the HNKTLMP group (P＜0.05). Cox regression analysis showed that LLMP and LNKTLMP were independent risk factors for patient mortality, with HR 4.620 and 2.706 (P＜0.05). The proportions of CD3+ CD4+ T and CD4+ /CD8+ T were higher in the LLMP group than those in the HLMP group (P＜0.05). The proportion of CD117+ in MM cells was higher in the LLMP group than that in the HLMP group (P＜0.05). Conclusion Patients with MM have abnormal secretion of LMP. The LMP and NKTLMP are closely correlated with prognosis of MM. Targeted regulation of LMP secretion may improve the survival and prognosis of MM. [ABSTRACT FROM AUTHOR]

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Copyright of Chinese Journal of Cancer Biotherapy is the property of Editorial Office of Chinese Journal of Cancer Biotherapy and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Chinese Journal of Cancer Biotherapy is the property of Editorial Office of Chinese Journal of Cancer Biotherapy and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

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Objective: To investigate the lipidomics differences of CD4+T immune cells in diabetic kidney disease (DKD) mice, and screen out the differential metabolites with biological significance. Methods: CD4 (L3T4) MicroBeads immunomagnetic beads were used to isolate CD4+T immune cells from spleen of BKS. Cg-Dock7m+/+ Leprdb/J mice with spontaneous DKD; the purity of CD4+T cells were identified by flow cytometry. The non-targeted lipidomics of CD4+T cells were detected by LC-MS/MS, and the differential metabolites were analyzed. Results: A total of 463 metabolites were detected by LC-MS. PCA and OPLS-DA analysis showed that the metabolic components were significantly separated; twenty-four differential metabolites were screened out. KEGG and enrichment analysis showed that the differential metabolites involved in the disorder of glycerol phospholipid metabolism. Conclusion: Phospholipid metabolism of CD4+T cells is closely related to the occurrence of DKD. Phospholipid metabolism targeting DKD CD4+T cells in DKD may be a new direction of DKD treatment. [ABSTRACT FROM AUTHOR]

Objective To evaluate the spermatogenesis functions of bilateral testes in rats after unilateral testicular torsion operation. Methods Total of 32 male Sprague-Dawley rats were randomly divided into four groups, and 8 rats in each group. For the rats in Group 1: a sham operation to the left testis was performed. For the rats in Group 2 and Group 3: I/R injury was created by rotating the left testis 720° in a clockwise direction for 2 h or 12 h then detorsion. For the rats in Group 4: the left testis rotated 720° for 12 h was performed through incision respectively. On 30 days after operation, the bilateral testes were collected and weighted, and their normal and apoptotic cells proportion were analyzed by the flow cytometry (FCM). Results Torsion/detorsion can significantly decreased the weight of ipsilateral testis and the percentage of normal cells, but increased the percentage of apoptotic cells and necrotic cells in Group 2 and Group 3. There were significant differences in the above indicators between Group 1 and Group 2/3 (P＜0.05). There were no significant differences in the weight or the percentage of normal cells of contralateral testis between Group 2 and Group 1 (P＞0.05), but significant differences in the percentage of apoptotic cells and necrotic cells (P＜0.05). The weight of contralateral testis was lower in Group 3 than that in Group 1 (P＜0.05). There were also significant changes in the percentage of normal cells, apoptotic cells and necrotic cells between Group 3/4 and Group 1 (P＜0.05). Compared with Group 3, the orchiectomy treatment in Group 4 increased the percentage of normal cells and decreased the percentage of apoptotic cells and necrotic cells (P＜0.05). Conclusion The short-time (2 h) testicular torsion/detorsion induce a continuous spermatogenesis damage for bilateral testis, but the spermatogenesis functions of bilateral testis, especially contralateral testis, can be gradually recovered. Therefore, surgical detorsion can provide even greater benefits. The long-time (12 h) testicular torsion/detorsion perhaps cause an unrecoverable injury to the bilateral testis. Whenever the torsional testis necrosis occurs definitely, the orchiectomy has beneficial effects against contralateral testis injury probably. [ABSTRACT FROM AUTHOR]

Copyright of Chinese Journal of Cancer Biotherapy is the property of Editorial Office of Chinese Journal of Cancer Biotherapy and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)