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.

Objective:To study the influence of methyltransferases like 3(METTL3)on the radiosensitivity of oral squamous cell carcinoma cells(OSCCs).Methods:The apoptosis level of OSCCs CAL27,SCC9 and SCC15 treated with X-ray radiation doses of 2,4 and 8 Gy respectively was compared by flow cytometry,the expression of methylated gene RNA and protein in the cells were examined by qRT-PCR and Western blot.m6A in the cells was quantified by LC/LC-MS method.qRT-PCR and Western blot were used to investigate the expression of methylated gene RNA and protein in the cells.Flow cytometry was used to examine the cell apoptosis level of CAL27 and SCC15 cells treated with METTL3 overexpression and knockdown respectively.The clone forma-tion of CAL27 and SCC15 cells after knockdown and overexpression of target genes followed by radiation treatment was observed by clonogenic assays.Results:The apoptosis rate of all the cell lines increased with the increase dose of radiation at each dose,CAL27 cells showed the highest and SCC15 showed the lowerst apoptosis rate.The RNA and protein expression levels of METTL3 in CAL27 were significantly lower than those of SCC15.m6A quantification showed that the methylation modification in CAL27 cells was lower than that in SCC15.The expression of METTL3 was increased in CAL27 and SCC15 cells after radiation treatment.Knockdown of METTL3 increaced the apoptosis rate and decreased the clonogenesity,overession of METTL3 the decreaced the ap optosis rate and increased the clonogenecity of the cells.Conclusion:Regulation of METTL3 can affect the radiotherapy sensitivity of OSCCs,METTL3 may become a new target for radiosensitization of OSCCs.

Objective To establish a 103-plex InDel-SNP composite marker amplification system based on Ion Torrent PGM/S5 sequencing platform targeting fragment length less than 175bp,providing an efficient analytical tool for forensic individual identification and paternity identification.Methods Using the human genome GRCh38 as the reference sequence,InDel-SNP composite marker loci suitable for Chinese population were screened from 22 pairs of autosomes and sex chromosomes by using dbSNP database with East Asian population as the target population.Population genetic data of 267 unrelated individuals of Han nationality in Shandong province were investigated,and genetic relationship was verified using 16 real triplet families.Results The 103-plex InDel-SNP composite marker detection system developed in this study,based on the second-generation sequencing platform,exhibits notable features including high sensitivity,excellent stability,strong specificity,and wide applicability to conventional test materials.It effectively enables the determination of typing results for degraded and mixed samples.Conclusion All 16 parent triplet pairs demonstrated adherence to Mendelian inheritance law without any observed allelic dropout or mutation.This detection system holds significant value for forensic individual identification and paternity testing.

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.

BackgroundPreventing and intervening in adolescent gaming disorder is of significant practical importance. Gaming motivation is strongly linked to gaming addiction and serves a key function in comprehending and addressing addictive gaming behaviors. However， the relationship between components of gaming motivation and symptoms of gaming disorder remains unclear. ObjectiveTo explore the relationship between components of gaming motivation and symptoms of gaming disorder among adolescents， so as to provide references for the prevention and intervention of gaming disorder in this population. MethodsFrom January to February 2024， a cluster sampling method was employed to select 1 414 adolescents from four middle schools in Sichuan Province and Chongqing Municipality as participants in the study. Online Game Motivation Scale （OGMS） and Gaming Disorder Scale for Adolescents （GADIS-A） were administered. Network analysis methods were utilized to investigate the relationships between components of gaming motivation and symptoms of gaming disorder. ResultsThe network edge weights revealed that achievement motivation was positively correlated with impaired game control ability， continued gaming despite negative consequences and the frequency of symptom occurrence （r=0.115， 0.050， 0.076， P<0.05）. Social motivation was negatively correlated with negative consequences （r=-0.054， P<0.05），while immersion motivation was positively correlated with continued gaming despite negative consequences （r=0.032， P<0.05）. Achievement motivation exhibited the highest strength centrality （1.157） among the three components of gaming motivation. ConclusionThe connections between components of gaming motivation and symptoms of gaming disorder exhibit distinct patterns， with each motivational component influencing gaming disorder through specific symptom pathway. Among these components， achievement motivation plays the most critical role in the interplay between gaming motivation and symptoms of gaming disorder. ［Funded by Chongqing Science and Health Joint Medical Science and Technology Innovation Projects General Projects （number， 2023MSXM133）］

Objective:To study the influence of methyltransferases like 3(METTL3)on the radiosensitivity of oral squamous cell carcinoma cells(OSCCs).Methods:The apoptosis level of OSCCs CAL27,SCC9 and SCC15 treated with X-ray radiation doses of 2,4 and 8 Gy respectively was compared by flow cytometry,the expression of methylated gene RNA and protein in the cells were examined by qRT-PCR and Western blot.m6A in the cells was quantified by LC/LC-MS method.qRT-PCR and Western blot were used to investigate the expression of methylated gene RNA and protein in the cells.Flow cytometry was used to examine the cell apoptosis level of CAL27 and SCC15 cells treated with METTL3 overexpression and knockdown respectively.The clone forma-tion of CAL27 and SCC15 cells after knockdown and overexpression of target genes followed by radiation treatment was observed by clonogenic assays.Results:The apoptosis rate of all the cell lines increased with the increase dose of radiation at each dose,CAL27 cells showed the highest and SCC15 showed the lowerst apoptosis rate.The RNA and protein expression levels of METTL3 in CAL27 were significantly lower than those of SCC15.m6A quantification showed that the methylation modification in CAL27 cells was lower than that in SCC15.The expression of METTL3 was increased in CAL27 and SCC15 cells after radiation treatment.Knockdown of METTL3 increaced the apoptosis rate and decreased the clonogenesity,overession of METTL3 the decreaced the ap optosis rate and increased the clonogenecity of the cells.Conclusion:Regulation of METTL3 can affect the radiotherapy sensitivity of OSCCs,METTL3 may become a new target for radiosensitization of OSCCs.

Objective To establish a 103-plex InDel-SNP composite marker amplification system based on Ion Torrent PGM/S5 sequencing platform targeting fragment length less than 175bp,providing an efficient analytical tool for forensic individual identification and paternity identification.Methods Using the human genome GRCh38 as the reference sequence,InDel-SNP composite marker loci suitable for Chinese population were screened from 22 pairs of autosomes and sex chromosomes by using dbSNP database with East Asian population as the target population.Population genetic data of 267 unrelated individuals of Han nationality in Shandong province were investigated,and genetic relationship was verified using 16 real triplet families.Results The 103-plex InDel-SNP composite marker detection system developed in this study,based on the second-generation sequencing platform,exhibits notable features including high sensitivity,excellent stability,strong specificity,and wide applicability to conventional test materials.It effectively enables the determination of typing results for degraded and mixed samples.Conclusion All 16 parent triplet pairs demonstrated adherence to Mendelian inheritance law without any observed allelic dropout or mutation.This detection system holds significant value for forensic individual identification and paternity testing.

Objective To investigate the regulatory role of nuclear factor erythroid 2-related factor 2(Nrf2)in the activation and polarization of microglia induced by methamphetamine(METH).Methods BV2 and HMC3 cells were studied in vitro and wild-type mice and Nrf2-knockout mice were studied in vivo.In vivo and in vitro toxicity models induced by METH were established,respectively.The activation and polarization of microglia in each group were examined using immunofluorescence and Western blot,respectively.Results METH treatment significantly increased the fluorescence level of inducible nitric oxide synthase(iNOS)in BV2 and HMC3 cells(P＜0.001),and significantly decreased the fluorescence level of Arginase1(Arg1)(P＜0.05,P＜0.01).METH exposure activated microglia in the cortex,increased expression levels of ionized calcium binding adaptor molecule-1(IBA1)and iNOS(P＜0.001,P＜0.05),and decreased the expression of Arg1(P＜0.01).The number of activated microglia was significantly increased after Nrf2 gene knockout(P＜0.01),compared with the WT METH group,while the expression levels of IBA1 and iNOS were also increased(P＜0.001,P＜0.01)and the expression level of Argl was decreased(P＜0.01).Conclusions Nrf2 plays an important role in regulating the activation and polarization of microglia induced by METH.Nrf2 may thus be a potential target for the treatment of neuroinflammation induced by METH.

Objective To investigate the effects of intraoperative fraction of inspiration oxygen(FiO2)30%and 80%on pulmonary function after laparoscopic radical prostatectomy for prostate cancer in elderly patients.Methods Sixty elderly patients,aged≥65 years,BMI 18-30 kg/m2,ASA physical sta-tus Ⅱ or Ⅲ,underwent elective laparoscopic radical prostatectomy for prostate cancer were selected.The patients were divided into two groups:the FiO2 30%group(group L)and the FiO2 80%group(group H),30 patients in each group.After tracheal intubation,the patients were manoeuvred,and inhaled oxygen con-centration was adjusted by 30%until the removal of the tracheal tube in group L.In group H,inhaled oxy-gen concentration was adjusted by 80%until the removal of the tracheal tube.HR and MAP were recorded 5 minutes after admission(T0),1 hour after the start of surgery(T1),2 hours after the start of surgery(T2),and 30 minutes after extubation(T3),arterial blood gas analysis was performed to record PaO2 and PaCO2,and the oxygenation index(OI)was calculated.Electrical impedance tomography(EIT)monitoring was performed at T0 and T3 to assess the pulmonary ventilation function,and the percentage of the area of the central ventilation zone(CoV),dependent static zone(DSS),and non-dependent static zone(NSS)were recorded.Exertional expiratory volume in the first second(forecd expirtory volume in the first second,FEV1),exertional lung capacity(forecdvital cipitory,FVC),and 1-second rate(FEV1/FVC)were meas-ured on preoperative day 1,postoperative day 1,postoperative day 3 and postoperative day 5.The occur-rence of pulmonary complications such as atelectasis,respiratory infection,and pleural effusion within 5 days postoperatively were recorded.Results Compared with group H,the percentage of DSS area at T3 was significantly decreased(P＜0.05),PaO2 and OI were significantly increased(P＜0.05),FVC,FEV1,and FEV1/FVC on postoperative day 1 were significantly increased(P＜0.05),FEV1/FVC on postopera-tive day 3 was significantly increased(P＜0.05),the incidence of cumulative pulmonary atelectasis was significantly decreased in group L within 5 days postoperatively(P＜0.05).Conclusion Compared with FiO2 80%,FiO2 30%intraoperatively significantly improves pulmonary ventilation and oxygenation 30 mi-nutes after laparoscopic radical prostatectomy with early postoperative lung function in elderly patients,and reduces postoperative pulmonary atelectasis.

Objective To explore the mediating effect of facial emotion recognition on prosocial behavior of medical students in mask-obscured scenes.Methods Fifty-three medical students from a medical university in Chongqing were enrolled from July to September 2023 to complete the facial emotion recognition task,the dictator gaming task and the state empathy test.Spearman correlation analysis was used to examine the correlation between mask wearing and state empathy,trait empathy and prosocial behaviours,and the PROCESS procedure was used to test the mediation of state empathy and the moderating effect of mask wearing or not.Results ①mask wearing,state empathy and prosocial behaviour were significantly correlated(P＜0.01);② State empathy exerted mediated effect between facial emotion recognition and prosocial behavior,with the largest effect size(47％)for the relative mediating effect of sadness;③The interaction terms of facial emotion recognition and mask wearing had a significant effect on state empathy(P＜0.05).Conclusion Facial emotion recognition can influence prosocial behavior directly and also exert indirect effect on prosocial behavior through state empathy.Compared to the condition without mask wearing,mask wearing can significantly facilitate the effect of happy,sad and neutral emotions on state empathy.