BACKGROUND: Cluster of differentiation 8 positive (CD8 + ) T cells play a crucial role in the response against tumors, including hepatocellular carcinoma (HCC), where their dysfunction is commonly observed. While the association between elevated peroxisome proliferator-activated receptor alpha (PPARα) expression in HCC cells and exosomes and unfavorable prognosis in HCC patients is well-established, the underlying biological mechanisms by which PPARα induces CD8 + T cell exhaustion mediated by HCC exosomes remain poorly understood. METHODS: Bioinformatics analyses and dual-luciferase reporter assays were used to investigate the regulation of microRNA-27b-3p ( miR-27b-3p ) and thymocyte selection-associated high mobility group box ( Tox ) by Pparα . In vitro and in vivo experiments were conducted to validate the effects of HCC-derived exosomes, miR-27b-3p overexpression, and Pparα on T cell function. Exosome characterization was confirmed using transmission electron microscopy, Western blotting, and particle size analysis. Exosome tracing was performed using small animal in vivo imaging and confocal microscopy. The expression levels of miR-27b-3p , Pparα , and T cell exhaustion-related molecules ( Tox , Havcr2 , and Pdcd1 ) were detected using quantitative reverse transcription polymerase chain reaction analysis, Western blotting analysis, immunofluorescence staining, and flow cytometry analysis. RESULTS: Pparα expression was significantly increased in HCC and negatively correlated with prognosis. It showed a positive correlation with Tox and a negative correlation with miR-27b-3p . The overexpressed Pparα from HCC cells was delivered to CD8 + T cells via exosomes, which absorbed miR-27b-3p both in vitro and in vivo , acting as "miRNA sponges". Further experiments demonstrated that Pparα can inhibit the negative regulation of Tox mediated by miR-27b-3p through binding to its 3'untranslated regions. CONCLUSIONS HCC-derived exosomes deliver Pparα to T cells and promote CD8 + T cell exhaustion and malignant progression of HCC via the miR-27b-3p /TOX regulatory axis. The mechanisms underlying T-cell exhaustion in HCC can be utilized for the advancement of anticancer therapies.
MicroRNAs/metabolism* ; PPAR alpha/genetics* ; Carcinoma, Hepatocellular/genetics* ; Humans ; Liver Neoplasms/genetics* ; CD8-Positive T-Lymphocytes/immunology* ; Exosomes/metabolism* ; Animals ; Cell Line, Tumor ; Mice ; High Mobility Group Proteins/genetics* ; Male ; T-Cell Exhaustion

Objective:To evaluate the diagnostic value of intestinal regional oxygen saturation（rSO 2）and fecal calprotectin in the occurrence and severity of necrotizing enterocolitis（NEC）in premature infants. Methods:A prospective observational study was conducted among premature infants admitted to Quanzhou Children's Hospital from October 2019 to December 2022. Intestinal rSO 2 was monitored within two hours of diagnosis of NEC，and fecal calprotectin was measured. Results:A total of 60 patients were included, including 30 cases with NEC and 30 cases without NEC, 14 cases of medical NEC, 16 cases of surgical NEC, and eight infants died due to NEC. Infants with NEC had lower intestinal rSO 2 [49（30，60）% vs. 66（60，69）%] and higher calprotectin levels [479（297，886）μg/g vs. 203（113，275）μg/g] than those in infants without NEC ( P<0.01). The levels of intestinal rSO 2 were lower in surgical NEC than those in medical NEC，and were lower in the death group than that in the survival group ( P<0.01)，but no similar difference was found in the levels of calprotectin. ROC curve analysis showed that intestinal rSO 2 combined with calprotectin had a sensitivity of 73%，a specificity of 100%，and the largest area under curve of 0.91 in the diagnosis of NEC. Intestinal rSO 2 had an optimal cut-off value of 31% in predicting death in infants with NEC，with a sensitivity of 100%，a specificity of 95%，and an area under curve of 0.99. Conclusion:Intestinal rSO 2 and fecal calprotectin can effectively identify the presence of NEC，and their combined detection can improve the diagnostic efficiency. Intestinal rSO 2 is a good predictor of the severity of NEC，but not fecal calprotectin.

OBJECTIVE: To explore the bidirectional regulation of acupuncture based on a subgroup analysis of multicenter randomized controlled trial of acupuncture with METHODS: A total of 519 patients were included in the analysis, including 137 patients with constipation type irritable bowel syndrome (IBS-C) (92 cases in the acupuncture group and 45 cases in the polyethylene glycol [PEG] group), and 382 patients with diarrhea type irritable bowel syndrome (IBS-D) (252 cases in the acupuncture group and 130 cases in the pinaverium group). The patients in the acupuncture group were given acupuncture at Baihui (GV 20), Yintang (GV 29), Tianshu (ST 25), Shangjuxu (ST 37), Zusanli (ST 36), Sanyinjiao (SP 6) and Taichong (LR 3) once every other day, 3 times a week. The patients in the PEG group received polyethylene glycol 4000 powder orally, and the pinaverium group received pinaverium bromide tablets orally. All were treated for 6 weeks. The IBS symptom severity score (IBS-SSS) was assessed at baseline, treatment period (2, 4, 6 weeks of treatment) and 12 weeks of follow-up, and the IBS quality of life (IBS-QOL) score was evaluated at the baseline period, 6 weeks of treatment and 12 weeks of follow-up. RESULTS: The total IBS-SSS scores of the two groups of IBS-C patients at 2, 4, 6 weeks of treatment and follow-up of 12 weeks were lower than those in the baseline period ( CONCLUSION Acupuncture with
Acupuncture Points ; Acupuncture Therapy ; Diarrhea ; Humans ; Irritable Bowel Syndrome/therapy* ; Quality of Life ; Treatment Outcome

Objective To summarize the therapeutic effects of extracorporeal membrane oxygenation (ECMO) on critically ill children with severe cardiopulmonary failure.Methods The pediatric patients supported with ECMO admitted to pediatric intensive care unit (PICU) from December 2015 to August 2017 were enrolled in this study.The data of demographics of patients,diagnosis,indication for ECMO,the procedure of ECMO support,complications,and survival status were analyzed.Results A total of 17 pediatric patients including 9 male and 8 female with severe cardiopulmonary failure treated with ECMO were studied.The median of age was 24 (2,117) months,and the median of body weight was 12 (5,33) kg.The indications for initiation of ECMO were cardiovascular failure with poor response to conventional therapy and severe acute respiratory distress syndrome (ARDS) without any beneficial effect obtained from mechanical ventilation.The percutaneous cannulation was done under ultrasound guidance by a team of trained intensivists through right cervical vein and internal carotid artery resulting in veno-arterial extracorporeal membrane oxygenation support.The mean duration of ECMO support was 212.5 h with median 188.5 (3-924) h.All patients were treated with mechanical ventilation for prevention from pulmonary atelectasis.Of 17 patients,10 were coupled with continuous renal replacement therapy (CRRT) to keep fluid balance.As a result,11 children (64.7％) were successfully weaned from ECMO defined as survival for 24 h after ECMO,and 10 children (58.8％) were alive to discharge from hospital.One of 2 cardiac arrest cases treated with extracorporeal cardiopulmonary resuscitation (ECPR) was alive.During ECMO support,there were 41 adverse events happened including pressure ulcers (64.7％),thrombocytopenia (52.9％),bleeding (35.3％).Conclusions The survival rate at discharge was 58.8％ in pediatric patients with severe cardiopulmonary failure with poor response to conventional therapy.Our data suggest that ECMO support is an important rescue technique for pediatric critical illness.

Objective To explore the influences of the final concentration and adding time of Cytochalasin-B (Cyt-B) on radiation-induced nucleoplasmic bridges (NPB) in cytokinesis-block assay.Methods Hunan peripheral blood samples were divided into 5 final concentration groups (group 2,4,6,8,10 μg/ml) according to different final concentrations of Cyt-B.Moreover,blood samples were divided into 4 adding time groups (group 0,28,40,44 h) according to different adding times of Cyt-B.Blood samples were irradiated with 0 (sham irradiation) and 2 Gy 60Co-rays in vitro,at a dose rate of 1 Gy/min.A cytokinesis-block assay was carried out to prepare NPB samples.The percentages of mononucleated,binucleated and multinucleated cells,as well as the frequencies of NPB and micronucleus (MN) in binucleated cells were analyzed using an optical microscope.Results Nuclear division index (NDI) and the percentages of binucleated cells increased with increased concentration of Cyt-B,and decreased with delayed adding time of Cyt-B (except group 0 h) in both final concentration groups and adding time groups.After exposed to 2 Gy,NPB frequencies were no significant difference (except group 0 h).MN frequencies had the trend of decreased with the increased concentration of Cyt-B,but no significant difference with adding time of Cyt-B.Conclusions In cytokinesis-block assay,different final concentration and adding time of Cyt-B may induce to the variation of NPB frequencies,but there was no significant difference.Appropriate increased final concentration or ahead adding time of Cyt-B can increase the percentage of binucleated cells that help to improve the efficiency of analysis.

Objective To investigate the dose response of S100A4 gene expression in the irradiated lymphoblastoid cells AHH-1 at different time points post irradiation.Methods AHH-1 cells was exposed to different doses(0,1,3,5,8,10,15 and 18 Gy)of 60Co γ-rays,and its mRNA levels of S100A4 was detected by reverse transcription PCR and real-time PCR at 4,8,12,24,48 and 72 h after irradiation.Results Within the range of applied doses,the level of S100A4 gene expression was upregulated with a good dose-response (R2 =0.79-0.93,P ＜ 0.05) and had obvious difference at different time points (F =8.91,P ＜ 0.01).Conclusion S100A4 gene expression at transcriptional level could be detected easily and had optimum dose-responses at certain time points after irradiation,and hence is applicable as a dosimeter.

Objective For developing safe and effective anti-radiation new drugs,the effects of different lipoic acid amino acid salts on radiosensitivity were investigated.Methods The free radical scavenging ability of the above salts was evaluated in Fenton system.CCK-8 assay and flow cytometry assay were performed to evaluate the survival rate of L02 and apoptosis of AHH-1 after γ-ray irradiation,respectively.Results The lipoic acid arginine salt had the best ability of scavenging free radicals in Fenton system with an IC50 of 8.40 μ moL/ml.The survival assay showed that lipoic acid amino acid salts had better stability and equal ability in radioprotection (P ＞ 0.05) compared with lipoic acid.The apoptosis assay indicated that all lipoic acid amino acid salts could inhibit radiation-induced apoptosis,where lipoic acid arginine salt was more effective (t =-6.67,P ＜ 0.01).Conclusions Lipoic acid arginine salt has good radioprotection effect on L02 and AHH-1 cells by scavenging free radicals.

Objective To investigate the dose-effect relationship of mRNA level of sensitive mitochondrial genes in human lymphoblastoid cells induced by ionizing radiation.Methods Seven human sensitive genes,including ND3,Cyt b,COX Ⅰ,COX Ⅱ,COX Ⅲ,ATPase6 and ATPase8 were chosen.Changes of mRNA level of these genes were detected by RT-PCR and Real-Time PCR at 24 h after irradiation in human lymphoblastoid cells,which were exposed to 0 - 15 Gy of 60 Co γ-rays.Results The expression of these 7 genes at mRNA level was up-regulated 24 h after irradiation in human lymphoblastoid cells.The level of gene expression of COX Ⅰ,which belongs to complex Ⅳ of mitochondrial respiratory chain,was most obvious,and the peak occurred after irradiation of 8 Gy,which was 13 times of the control group.A good dose-effect relationship was showed for COX Ⅲ gene expression at dose range of 3 -10 Gy as well as 3 - 15 Gy for other 3 genes including ND3,ATPase6 and ATPase8.Conclusions Gene expression levels of COX Ⅲ,ND3,ATPase6 and ATPase8 24h post-irradiation at certain irradiation dose range could be used for radiation damage biomarkers.

Objective To explore the changes of human mitochondrial COX Ⅰ , COX Ⅱ and COX Ⅲ genes expression induced by ionizing irradiation. Methods Changes of human COX genes expression were detected by RT-PCR and Real-time PCR 8 h after the irradiation in human lymphoblastoid cell lines,which were exposed to 1-10 Gy60Co γ-rays. The protein levels were detected by flow cytometry and the COX activity was measured by colorimetry. The dose-effect relationships between the expression changes of the genes and the doses were established. The changes of these genes expression were also analyzed at different post-radiation time-points between 0. 5 h and 72 h after irradiation of 5 Gy in order to explore the time-effect. Results The expression of 3 genes at mRNA level was up-regulated. A good dose-effect relationship was showed for COXⅠ and COX Ⅲ at dose range of 0-3 Gy and 0-8 Gy for COX Ⅱ ( F COXⅠ=116. 62, FCOXⅡ = 17. 89, FCOXⅢ = 8.20, P ＜ 0. 05). For the time-effect after irradiation, the gene expression levels of COX Ⅱ and COX Ⅲ genes were up-regulated and the peak change occurred at 4 h after irradiation. For COX Ⅰ gene, the mRNA expression levels were down-regulated during 0.5-72 h( FCOXⅠ =31.99, FCOXⅡ = 19.47, FCOXⅢ = 20. 64, P ＜0. 05 ). At the protein level, the levels of COX Ⅰ and COX Ⅱ were lowered in lower doses and enhanced in higher doses, and the levels of COX Ⅲ were decreased at all dose levels (FCOX Ⅰ = 16.96, FCOXⅡ = 32.5, FCOXⅢ = 6. 51, P ＜ 0. 05 ). The protein levels of COX Ⅰ and COX Ⅱ were enhanced during 4-72 h and 8-72 h respectively after 5 Gy irradiation ( FCOX Ⅰ = 14.68,FCOXⅡ = 17. 18, FCOXⅢ =2. 52, P ＜0. 05). The activities of COX were lowered at different dose levels and different time-points. Conclusions Ionizing radiation might induce the changes in mitochondrial COX Ⅰ,COX Ⅱ and COX Ⅲ gene expression, and lead to the reduction of the COX activities.