1.Society of Critical Care Medicine 2024 Guidelines on Adult ICU Design: An Interpretation
Hui ZHANG ; Jianhua SUN ; Wanchen ZHAO ; Lingli XIE ; Cong MA ; Yifan FANG ; Jing CAI ; Na GUO
Medical Journal of Peking Union Medical College Hospital 2026;17(2):421-428
This article provides a systematic interpretation and review of the
2.Mechanism of Guihuang Formula in Regulating NLRP3 Inflammasome-mediated Pyroptosis in Treatment of Chronic Prostatitis
Qinghe GAO ; Jianhua FU ; Shengjing LIU ; Ziwei ZHAO ; Ming ZHAO ; Boda GUO
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(2):108-116
ObjectiveTo observe the mechanism of Guihuang formula in regulating the activation of NOD-like receptor protein 3 (NLRP3) inflammasome and inhibiting pyroptosis in the treatment of type Ⅲ prostatitis. Methods(1) In an animal experiment, 50 Sprague Dawley (SD) rats were randomly divided into a blank group, a model group, and low-dose, medium-dose, and high-dose groups of Guihuang formula, with 10 rats in each group. Except for the blank group, the type Ⅲ prostatitis rat model was prepared for the other four groups.After the modeling was successful, the blank group and the model group were given normal saline intragastrically, and the low-dose, medium-dose, and high-dose groups of Guihuang formula were given intragastrically with Guihuang formula (4.9, 9.8, 19.6 g·kg-1). After 30 days of intragastrical administration, samples were taken for detection. Inflammatory cell infiltration in prostate tissue was observed by hematoxylin-eosin (HE) staining, and serum IL-1β and IL-18 levels were measured by enzyme-linked immunosorbent assay (ELISA). Serum malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione peroxidase (GSH-Px) levels were determined by biochemistry. NLRP3 expression in prostate tissue was assessed by immunohistochemistry, and the expression of NLRP3, cysteine-aspartic acid protease-1 (Caspase-1), and gasdermin D (GSDMD) in prostate tissue was measured by Western blot. (2) In a cell experiment, human normal prostate epithelial cells (RWPE-1 cells) were divided into a blank group, a model group, a Guihuang formula group, and an NLRP3 inhibitor group (MCC950 group). Except for the blank group, the other three groups were stimulated by 100 μg·L-1 lipopolysaccharide (LPS) for 4 h and 5 mol·L-1 adenosine triphosphate (ATP) for 30 min to prepare the pyroptosis model. After successful modeling, blank serum was given to the blank group and the model group. 6.25 μg·mL-1 Guihuang formula drug-containing serum was added to the Guihuang formula group, and MCC950 was added to the MCC950 group on the basis of the model group. Propidium iodide (PI) uptake and Caspase-1 expression were detected by flow cytometry, and lactate dehydrogenase (LDH) level in the cell supernatant was measured by biochemistry. Interleukin (IL)-1β and IL-18 levels of the cell supernatant were determined by ELISA, and the expression of NLRP3, Caspase-1, and GSDMD was detected in Western blot. Results(1) For the animal experiment, compared with the blank group, the model group showed significant infiltration of inflammatory cells in prostate tissue, while the low-dose, medium-dose, and high-dose groups of Guihuang formula showed reduced infiltration of acinar inflammatory cells, reduced degree of glandular epithelial degeneration and interstitial edema, and significantly reduced degree of damage. Compared with those in the blank group, the levels of IL-1β and IL-18 in the serum of the model group were significantly increased (P<0.01). Compared with the model group, the low-dose, medium-dose, and high-dose groups of Guihuang formula showed a significant decrease in serum IL-1β and IL-18 levels (P<0.01). Compared with that in the blank group, the serum MDA level in the model group significantly increased (P<0.01). Compared with that in the model group, the MDA level in the low-dose, medium-dose, and high-dose groups of Guihuang formula was significantly reduced (P<0.01). Compared with those in the blank group, the levels of SOD and GSH-Px in the serum of the model group significantly decreased (P<0.05). Compared with the model group, the low-dose, medium-dose, and high-dose groups of Guihuang formula showed a significantly increase in SOD (P<0.01). Compared with the model group, the low-dose, medium-dose, and high-dose groups of Guihuang formula showed a significantly increase in GSH-Px (P<0.05). Immunohistochemistry showed that compared with the blank group, the model group had high expression of NLRP3 molecule in prostate tissue. The expression of NLRP3 in the low-dose, medium-dose, and high-dose groups of Guihuang formula was significantly lower than that in the model group. Compared with those in the blank group, the expression levels of NLRP3, Caspase-1, and GSDMD proteins in the prostate tissue of the model group were significantly increased (P<0.01). Compared with those in the model group, the expression levels of NLRP3, Caspase-1, and GSDMD proteins in the low-dose, medium-dose, and high-dose groups of Guihuang formula were significantly inhibited (P<0.01). (2) For the cell experiment, compared with that in the blank group, the PI uptake rate of RWPE-1 cells in the model group significantly increased (P<0.01). Compared with that in the model group, the PI uptake rate of the Guihuang formula group and the inhibitor group significantly decreased (P<0.01). Compared with that in the blank group, the expression of Caspase-1 in the model group was significantly higher (P<0.01). Compared with that in the model group, the Caspase-1 in the Guihuang formula group and the inhibitor group significantly decreased (P<0.01). Compared with the blank group, the model group showed an increase in LDH release (P<0.01). Compared with the model group, the Guihuang formula group and the inhibitor group showed a significantly decrease in LDH release (P<0.01). Compared with those in the blank group, the levels of IL-1β and IL-18 in the supernatant of the model group were significantly increased (P<0.01). Compared with the model group, the Guihuang formula group and the inhibitor group showed a significantly decrease in the levels of IL-1β and IL-18 (P<0.01). Compared with those in the blank group, the expression levels of NLRP3, Caspase-1, and GSDMD proteins significantly increased in the model group (P<0.01). Compared with those in the model group, the protein expression levels of NLRP3, Caspase-1, and GSDMD were significantly reduced in the Guihuang formula group and inhibitor group (P<0.01). ConclusionGuihuang formula can inhibit the activation of Caspase-1, prevent GSDMD cleavation and lysis, and inhibit cell pyrodeath in the treatment of type Ⅲ prostatitis by inhibiting the activation of NLRP3 inflammasome.
3.Study on the effects and mechanisms of Lycium ruthenicum Murr. in improving sleep
Ming QIAO ; Yao ZHAO ; Yi ZHU ; Yexia CAO ; Limei WEN ; Yuehong GONG ; Xiang LI ; Juanchen WANG ; Tao WANG ; Jianhua YANG ; Junping HU
China Pharmacy 2026;37(1):24-29
OBJECTIVE To investigate the effects and mechanisms of Lycium ruthenicum Murr. in improving sleep. METHODS Network pharmacology was employed to identify the active components of L. ruthenicum and their associated disease targets, followed by enrichment analysis. A caffeine‑induced zebrafish model of sleep deprivation was established , and the zebrafish were treated with L. ruthenicum Murr. extract (LRME) at concentrations of 0.1, 0.2 and 0.4 mg/mL, respectively; 24 h later, behavioral changes of zebrafish and pathological alterations in brain neurons were subsequently observed. The levels of inflammatory factors [interleukin-6 (IL-6), IL-1β, IL-10, tumor necrosis factor-α (TNF-α)], oxidative stress markers [superoxide dismutase (SOD), malondialdehyde (MDA), glutathione peroxidase (GSH-Px), catalase (CAT)], and neurotransmitters [5- hydroxytryptamine (5-HT), γ-aminobutyric acid (GABA), glutamic acid (Glu), dopamine (DA), and norepinephrine (NE)] were measured. The protein expression levels of protein kinase B1 (AKT1), phosphorylated AKT1 (p-AKT1), epidermal growth factor receptor (EGFR), B-cell lymphoma 2 (Bcl-2), sarcoma proto-oncogene,non-receptor tyrosine kinase (SRC), and heat shock protein 90α family class A member 1 (HSP90AA1) in the zebrafish were also determined. RESULTS A total of 12 active components and 176 intersecting disease targets were identified through network pharmacology analysis. Among these, apigenin, naringenin and others were recognized as core active compounds, while AKT1, EGFR and others served as key targets; EGFR tyrosine kinase inhibitor resistance signaling pathway was identified as the critical pathway. The sleep improvement rates in zebrafish of LRME low-, medium-, and high-dose groups were 54.60%, 69.03% and 77.97%, 开发。E-mail:hjp_yft@163.com respectively, while the inhibition ratios of locomotor distance were 0.57, 0.83 and 0.95, respectively. Compared with the model group, the number of resting counts, resting time and resting distance were significantly increased/extended in LRME medium- and high-dose groups (P<0.05). Neuronal damage in the brain was alleviated. Additionally, the levels of IL-6, IL-1β, TNF-α, MDA, Glu, DA and NE, as well as the protein expression levels of AKT1, p-AKT1, EGFR, SRC and HSP90AA1, were markedly reduced (P<0.05), while the levels of IL-10, SOD, GSH-Px, CAT, 5-HT and GABA, as well as Bcl-2 protein expression, were significantly elevated (P<0.05). CONCLUSIONS L. ruthenicum Murr. demonstrates sleep-improving effects, and its specific mechanism may be related to the regulation of inflammatory responses, oxidative stress, neurotransmitter balance, and the EGFR tyrosine kinase inhibitor resistance signaling pathway.
4.Development and validation of the rapid health aging assessment scale for the Chinese population
Bingqi YE ; Jialu YANG ; Jianhua LI ; Wunong CHEN ; Jianhua YE ; Xiaotao ZHOU ; Yong WANG ; Siqi LI ; Qi ZHANG ; Wanying ZHAO ; Jiayi SONG ; Chun WANG ; Yan LIU ; Min XIA
Chinese Journal of Preventive Medicine 2025;59(7):1078-1083
Objective:To develop a rapid assessment scale for healthy aging suitable for the Chinese population.Methods:Based on existing healthy aging assessment scales, national standards, and expert consensus, an initial Healthy Aging Rapid Assessment Scale was drafted through two rounds of expert consultation. A pre-survey was conducted with 3 220 subjects recruited from Guangzhou between July 2023 and July 2024. Items were screened through item analysis and exploratory factor analysis to form the final scale. Reliability and validity of the final scale were validated across five cities: Guangzhou, Dongguan, Shenzhen, Baoding, and Chuxiong.Results:The initial version comprised 36 items, while the finalized scale contained 18 items across three dimensions: metabolic health, mental health, and cognitive health. Test-retest reliability ranged from 0.71 to 0.81 across all study sites. The Spearman-Brown coefficient varied between 0.91-0.96, Cronbach′s α between 0.77-0.83, comparative fit index (CFI) between 0.90-0.98, goodness-of-fit index (GFI) between 0.90-0.99, and root-mean-square error of approximation (RMSEA) between 0.03-0.09. For the three dimensions, reliability and validity metrics demonstrated consistency: Spearman-Brown coefficients 0.87-0.99, Cronbach′s α 0.77-0.83, CFI 0.90-0.98, GFI 0.90-0.99, and RMSEA 0.03-0.09 across four regions.Conclusion:The developed Healthy Aging Rapid Assessment Scale for the Chinese population exhibits robust reliability and validity.
5.Analysis of influencing factors for early neurological deterioration in isolated basal ganglia lacunar infarction
Journal of Apoplexy and Nervous Diseases 2025;42(1):42-46
摘要
目的 探讨孤立性基底节区腔隙性脑梗死早期神经功能恶化(END)的相关影响因素。方法 连续性收集2020年1月—2023年12月就诊于郑州大学人民医院的孤立性基底节区腔隙性脑梗死患者236例,临床资料完整,根据是否出现END将患者分为END组59例及非END组177例,比较患者一般资料,使用多因素二元Logistic回归分析基底节区腔隙性脑梗死患者发生END的影响因素。结果 急性孤立性基底节区腔隙性脑梗死END发生率为25%(59/236),END组病灶累及内囊后肢患者比例、入院NIHSS评分、HbA1c水平、收缩压、女性均高于非END组,两组比较差异有统计学意义(P<0.05)。多因素二元Logistic回归模型分析显示病灶累及内囊后肢(OR=3.167,95%CI 1.305~7.690,P=0.011)是END发生的独立危险因素,HbA1c水平(OR=6.368,95%CI 1.555~26.075,P=0.010)、入院NIHSS评分(OR=2.019,95%CI 1.236~3.299,P=0.005)、收缩压(OR=1.626,95%CI 1.373~1.926,P<0.001)是END发生的相关危险因素。结论 孤立性基底节区腔隙性脑梗死END发生率较高,与病灶累及内囊后肢、入院NIHSS评分高、HbA1c水平升高、收缩压高相关。
Abstract
Objective To identify influencing factors for early neurological deterioration (END) in isolated basal ganglia lacunar infarction (iBGLI). Methods Clinical data were continuously collected from 236 patients with iBGLI confirmed by magnetic resonance imaging between January 2020 and December 2023. The patients were divided into END group (n=59) and non-ED group (n=177) according to the presence or absence of END. General patient information was compared between the two groups, and factors influencing the occurrence of END in patients with iBGLI were identified by multivariate binary logistic regression. Results The incidence of END in acute iBGLI was 25% (59/236). The percentage of patients with lesions affecting the posterior limb of the internal capsule, admission NIHSS score, HbA1c level, systolic blood pressure, and number of females were significantly higher in the END group than in the non-END group (all P<0.05). Multivariate binary logistic regression showed that lesions affecting the posterior limb of the internal capsule (odds ratio (OR=3.167,95%CI 1.305~7.690,P=0.011) was an independent risk factor for the development of END,whereas HbA1c level(OR=6.368,95%CI 1.555~26.075,P=0.010), admission NIHSS score(OR=2.019,95%CI 1.236~3.299, P=0.005), and systolic blood pressure(OR=1.626,95%CI 1.373~1.926,P<0.001) were associated risk factors for END. Conclusion The higher incidence of END in iBGLI is associated with lesions affecting the posterior limb of the internal capsule, admission NIHSS score, HbA1c level, and systolic blood pressure.
Prognosis
6.Mining molecular biomarkers regulating the occurrence of kidney renal clear cell carcinoma based on bioinformatics methods
Feng GUO ; Chenyu WANG ; Zhenfeng SHI ; Jianhua ZHAO ; Wenlong FAN ; Kadeer AIHEMAITI ; Zecheng NI
Journal of Modern Urology 2025;30(3):215-222
Objective: To identify biomolecular markers closely related to the occurrence of kidney renal clear cell carcinoma (KIRC) and verify their expression levels in clinical samples. Methods: Stage Ⅰ KIRC mRNA sequencing data were obtained from The Cancer Genome Atlas (TCGA).Principal component analysis (PCA) was used for dimensionality reduction to screen differentially expressed genes (DEGs),which then underwent GO and KEGG analyses.Weighted gene co-expression network analysis (WGCNA) was used to screen genes significantly related to KIRC,and a protein-protein interaction (PPI) network was constructed to screen hub genes.The diagnostic value of hub genes was evaluated with receiver operating characteristic (ROC) curve,and their prognostic value was analyzed using survival curve plots.The correlation between the mRNA expressions of hub genes and the pathological stages of KIRC was analyzed.Clinical samples of 20 patients with stage Ⅰ KIRC treated in our hospital were included,and the expressions of the hub genes in cancerous and adjacent tissues were detected with reverse transcription real-time quantitative polymerase chain reaction (RT-qPCR),Western blotting,and enzyme-linked immunosorbent assay (ELISA). Results: A total of 8223 DEGs were screened out,including 4092 up-regulated ones and 4131 down-regulated ones.GO analysis showed that DEGs were related to bioadhesion,plasma membrane composition,and transporter activity.KEGG analysis showed that DEGs were related to pathways such as cell adhesion molecules,cytokine-cytokine receptor interactions,and interactions between viral proteins and cytokines and cytokine receptors.WGCNA analysis obtained 171 genes that were significantly related to stage Ⅰ KIRC.The hub gene,lymphocyte cytosolic protein 2 (LCP2),screened out by the PPI network,was significantly related to stage Ⅰ KIRC.The area under the ROC curve was 0.96.The expression level was negatively correlated with the overall survival rate of patients.The expression of LCP2 was related to the stage and lymph node metastasis.Clinical verification showed that the mRNA and protein relative expressions of LCP2 in KIRC tissues were significantly higher than those in adjacent tissues (P<0.000 1). Conclusion: LCP2 is significantly up-regulated in stage Ⅰ KIRC tissues and can be used as a potential biomarker for the early diagnosis and treatment of KIRC.
7.Research on the influence of CYP2A6 gene polymorphism on liver function injury in the treatment of hyperthyroidism with methimazole
Zuzhi ZHAO ; Haixia ZHAO ; Pengfei XU ; Yinghao FAN ; Peng SUN ; Suqin ZHANG ; Xinguang QIU ; Jianhua LI
Chinese Journal of Endocrine Surgery 2025;19(4):527-532
Objective:To investigate the influence of cytochrome P450 2A6 (CYP2A6) gene polymorphisms on liver function injury in patients with hyperthyroidism treated with methimazole.Methods:The study selected 90 patients with hyperthyroidism who were treated with methimazole in the Department of Thyroid Surgery at the First Affiliated Hospital of Zhengzhou University from Sept. 2023 to Aug. 2024 as the research subjects. Based on the occurrence of liver injury, they were divided into a liver injury group ( n=36) and a non-liver injury group (n=54). Peripheral blood DNA was extracted from the patients, and the CYP2A6 gene genotypes (rs8192725, rs8192720, and rs28399433) were detected using the polymerase chain reaction (PCR) amplification method. The association between CYP2A6 gene polymorphisms and liver injury induced by methimazole treatment in hyperthyroidism was analyzed. Results:The comparison of genotype distribution frequencies at the rs8192725 locus between the liver injury group and the non-liver injury group showed a statistically significant difference ( P<0.05). The AG and GG genotypes at the rs8192725 locus were protective factors against liver injury in patients with hyperthyroidism (AG vs. AA, OR: 0.21; 95% CI: 0.08-0.57; P<0.05; GG vs. AA, OR: 0.24; 95% CI: 0.06-0.89; P<0.05; AG+GG vs. AA, OR: 0.22; 95% CI: 0.09-0.54; P<0.05). The frequency of the G allele of rs8192725 in the liver injury group was significantly lower than that in the non-liver injury group (G vs. A, OR: 0.36; 95% CI: 0.19-0.70; P<0.05), indicating that it is a protective factor for liver injury in hyperthyroid patients receiving methimazole treatment. Conclusions:The CYP2A6 gene polymorphism at the rs8192725 locus is associated with the occurrence of liver injury in patients with hyperthyroidism treated with methimazole. The G allele may be a protective factor against liver injury in patients with hyperthyroidism, suggesting that individualized treatment plans can be developed based on the patient's genotype.
8.Correlation between serum cold-inducible RNA-binding protein, cystatin C and cognitive impairment in cerebral small vessel disease
Zhiming LI ; Lili ZHU ; Shengqi FU ; Baoyang SHI ; Jianhua ZHAO
Chinese Journal of Behavioral Medicine and Brain Science 2025;34(8):686-691
Objective:To explore the correlation between levels of serum cold-inducible RNA-binding protein(CIRBP), cystatin C(CysC) and cognitive impairment in patients with cerebral small vessel disease(CSVD), as well as the diagnostic value of CI in CSVD.Methods:A total of 90 CSVD patients admitted to the Neurology Department of Zhengzhou People's Hospital from January 2024 to December 2024 were consecutively selected. According to the mini-mental state examination(MMSE) and Montreal cognitive assessment(MoCA), they were divided into non cognitive impairment group(NCI group, n=47) and cognitive impairment group(CI group, n=43). The general clinical data of patients were collected.Fasting venous blood was collected in the morning on the second day of admission to measure serum CIRBP, CysC, homocysteine(Hcy) and high-sentivity C-reactive protein(hs-CRP) levels.A multiple-factor Logistic regression model was applied to identify independent risk factors for CI in CSVD patients, the predictive performance of the model was evaluated by receiver operating characteristic(ROC) curves, and the area under the curve(AUC) value was calculated to quantify the diagnostic accuracy of the model. Results:The levels of Hcy((13.01±4.22)μmol/L vs (11.44±3.00)μmol/L), hs-CRP((2.84±3.01)mmol/L vs (1.81±1.32)mmol/L), CIRBP((2 412.40±967.78)pg/mL vs (1 715.13±971.98)pg/mL), and CysC((1.93±1.08)mg/L vs (1.24±0.87)mg/L) in the CI group were significantly higher than those in the NCI group(all P<0.05). Multiple Logistic regression analysis showed that both CIRBP( OR=1.001, 95% CI=>1.000-1.002, P=0.011) and CysC( OR=1.833, 95% CI=1.056-3.181, P=0.031) were independently influencing factors of the occurrence of CI in CSVD patients(all P<0.05). ROC curve analysis showed that the optimal cutoff values for serum CIRBP and CysC levels to evaluate CI were 1 875.50 pg/mL and 1.42 mg/L, respectively. The AUC (95% CI) were 0.805(95% CI=0.713-0.897, P<0.001) and 0.716(95% CI=0.607-0.825, P<0.001), respectively.The AUC(95% CI) of combined detection of CIRBP+ CysC was 0.820(95% CI=0.733-0.907, P<0.001), with specificity and sensitivity of 89.4% and 67.4%. Conclusion:The serum CIRBP and CysC levels can serve as independent predictors of CI in CSVD patients. Combined testing can improve the accuracy of patient condition assessment and may assist in the diagnosis and prediction of cognitive impairment in CSVD.
9.The correlation between serum sTREM2 levels and cognitive impairment in patients with cerebral small vessel disease
Hanfang CUI ; Fangyuan DING ; Zhixiu XU ; Qing LI ; Yifan ZHANG ; Sen ZHANG ; Mengke GAO ; Yuhui CHEN ; Xiaowen ZHAO ; Jialu ZHAO ; Chengbiao LU ; Shaomin LI ; Jianhua ZHAO
Chinese Journal of Behavioral Medicine and Brain Science 2025;34(11):976-982
Objective:To explore the correlation between serum soluble triggering receptor expressed on myeloid cells 2 (sTREM2) and cognitive impairment in patients with cerebral small vessel disease (CSVD), and the role of deep medullary vein (DMV) score in this process.Methods:A total of 140 patients with CSVD admitted to the Department of Neurology, the First Affiliated Hospital of Henan Medical University from December 2022 to August 2024 were selected as the research objects. The basic data statistics, head magnetic resonance imaging examination, cognitive function assessment, serum sTREM2 detection and DMV score were performed. All data were analyzed by SPSS 29.0 software and GraphPad Prism 10.0 software packages. Logistic regression model was used to explore the influencing factors of cognitive impairment. Structural equation model was used to analyze the mediating effect of DMV score on the association between serum sTREM2 and cognitive impairment. Receiver operating characteristic (ROC) curve was used to evaluate the predictive value of serum sTREM2 level and DMV score for cognitive impairment in CSVD patients.Results:Serum sTREM2 level ( B=0.017, OR=1.017, 95% CI=1.003-1.031), DMV score ( B=0.375, OR=1.455, 95% CI=1.175-1.802) and years of education ( B=-0.248, OR=0.780, 95% CI=0.635-0.958) were risk factors for cognitive impairment (all P<0.05). sTREM2 not only directly affected cognitive function, but also indirectly affected cognitive function through DMV score. The direct effect (effect size=-0.022) and mediating effect (effect size=-0.007) accounted for 75.9% and 24.1% of the total effect (effect size=-0.029), respectively. The areas under the ROC curve of serum sTREM2 level, DMV score, and their combination for predicting cognitive impairment in CSVD patients were 0.880, 0.891, and 0.910, respectively (all P<0.001). Conclusion:Serum sTREM2 not only directly affects the cognitive function of patients with cerebral small vessel disease, but also indirectly affects cognitive function through DMV score. The combination of serum sTREM2 levels and DMV score has high predictive value for the risk of CSVD-related cognitive impairment.
10.Exosome Linc00665 regulates radiotherapy resistance in oral squamous cell carcinoma by regulating T cell immunoreactivity
Huan LI ; Junhong HUANG ; Yating HU ; Yahui LI ; Zihui YANG ; Zhenyan ZHAO ; Xinjie YANG ; Jianhua WEI
Journal of Practical Stomatology 2025;41(6):744-749
Objective:To investigate the function and mechanism of exosome Linc00665 in modulating CD8+T cell immunoreactivity to promote radiotherapy resistance in OSCC.Methods:HOEC,SCC9 and SCC9-RR exosomes were extracted and identified,and the expression of Linc00665 was detected by qRT-PCR in cell lines and exosomes.The expression of TNF-α,IFN-γ,perforin and granzyme B in each treatment group was detected by ELISA(PBS,SCC9 exo,SCC9-RR exo).The killing ability of CD8+T cells against SCC9 cells in each treatment group was detected by CCK-8 assay.The targets of Linc00665 were further bioinformatically ana-lyzed and verified by qRT-PCR and Western blot.The expression of Linc00665,miR-28-5p and PD-1 in CD8+T cells was exogenous-ly regulated,the expression of immunoreactive factors in the supernatants of each treatment group was detected by ELISA(NC,sh-Linc00665,miR-28-5p inhibitor,sh-PD-1),and the killing ability of cells in each group was detected by CCK-8 method.Results:The concentrations of TNF-α,IFN-γ,perforin and granzyme B in the supernatants of cell culture in the SCC9-RR exo/CD8+T group were significantly decreased compared with those in the PBS/CD8+T group and the SCC9 exo/CD8+T group(P<0.05),and the kill-ing ability of the cells in the SCC9-RR exo group was significantly decreased compared with those in the PBS group and the SCC9 exo group(P<0.05),suggesting that SCC9-RR exo could inhibit the tumor killing ability of CD8+T cells.qRT-PCR results suggested that Linc00665 was highly expressed in the SCC9-RR cell line as well as exosome(P<0.05).It was further verified by bioinformat-ics analysis that Linc00665 could regulate PD-1 expression via miR-28-5p,thereby modulating CD8+T cell immunoreactivity to pro-mote OSCC radiotherapy resistance.Conclusion:Exosome Linc00665 regulates CD8+T cell immunoreactivity through miR-28-5p/PD-1 axis to promote OSCC radiotherapy resistance.

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