Objective:To establish a mouse model of liver cancer resistant to PD-1 monoclonal antibody and analyze the changes in its metabolomics to explore the potential mechanism of drug resistance.Methods:BALB/c mice were randomly divided into control and treatment groups after being loaded with tumor,and a normal group was additionally set up.The normal and control groups were injected with saline,and the treatment group was injected with PD-1 monoclonal antibody,after which the mice in the treatment group were screened for drug resistant and response groups.Observed the drug-resistant situation,body mass,tumor growth and survival rate of mice in each group,calculate the spleen index.The pathological features of tumor tissues were observed by HE staining method.Serum metabolites were detected by non-targeted metabolomics.Finally,a bivariate Pearson correlation analysis was conducted between the differential serum metabolites and tumor size.Results:The tumor-bearing mouse model with PD-1 monoclonal antibody resistance was successfully established,and the drug resistance rate of the mice was 50％.Compared with the normal and response groups,mice in the resistant group showed an increase in body weight,a significant increase in tumor volume,a decrease in survival rate,and a significant increase in splenic index.There was less lymphocyte infiltration in the tumor tissue.Metabolomics analysis showed that the serum levels of glutamic acid and aspartic acid increased and malic acid decreased in the resistant mice compared with the response group,and these changes were closely related to the arginine biosynthesis pathway.Conclusions:The tumor-bearing mouse model with PD-1 monoclonal antibody resistance was successfully established.The changes in its peripheral serum metabolomics mainly involve arginine metabolism and the related changes of aspartate,malate and glutamate.

Objective To construct mice hepatocellular carcinoma models with different tumor immune microenvironments(TIME)and explore the differences.Methods H22 and hepa1-6 were used to construct subcutaneous transplantation tumor model of C57 mice as homologous hepatocellular carcinoma cell lines(denoted as H22 group and hepal-6 group,each n=8),and the differences of TIME were evaluated.Immunohistochemistry was used to detect and quantify the infiltration of T cells,CD4+T cells,CD8+T cells,regulatory T cells and B cells in TIME.Flow cytometry was performed to detect the differences of composition of immune cell subpopulations in peripheral blood and tumor parenchyma.Gene expression profile characteristics of tumor tissue were analyzed based on high-throughput transcriptome sequencing technology,and enrichment analyses of immune-related signaling pathways were evaluated combined with gene ontology(GO)and Kyoto encyclopedia of genes and genomes(KEGG).Results H22 group showed cold and hepa1-6 group showed hot TIME characteristics.The number of T cells,CD4+T cells and CD8+T cells in tumor tissue of H22 group were all lower,while the proportion of T cells,CD4+T cells and CD8+T cells in peripheral blood were all higher than those of hepa1-6 group(all P＜0.05).Compared with H22 group,up-regulated genes of tumor tissue in hepa1-6 group expressed significantly enriched in tumor immune activation-related signaling pathways.Conclusion H22 and hepa1-6 hepatocellular carcinoma models showed distinct TIME characteristics of cold and hot tumors,respectively,and the amount of immune cells in tumor tissue of the former were significantly lower than those in the latter.

Objective:To establish a mouse model of liver cancer resistant to PD-1 monoclonal antibody and analyze the changes in its metabolomics to explore the potential mechanism of drug resistance.Methods:BALB/c mice were randomly divided into control and treatment groups after being loaded with tumor,and a normal group was additionally set up.The normal and control groups were injected with saline,and the treatment group was injected with PD-1 monoclonal antibody,after which the mice in the treatment group were screened for drug resistant and response groups.Observed the drug-resistant situation,body mass,tumor growth and survival rate of mice in each group,calculate the spleen index.The pathological features of tumor tissues were observed by HE staining method.Serum metabolites were detected by non-targeted metabolomics.Finally,a bivariate Pearson correlation analysis was conducted between the differential serum metabolites and tumor size.Results:The tumor-bearing mouse model with PD-1 monoclonal antibody resistance was successfully established,and the drug resistance rate of the mice was 50％.Compared with the normal and response groups,mice in the resistant group showed an increase in body weight,a significant increase in tumor volume,a decrease in survival rate,and a significant increase in splenic index.There was less lymphocyte infiltration in the tumor tissue.Metabolomics analysis showed that the serum levels of glutamic acid and aspartic acid increased and malic acid decreased in the resistant mice compared with the response group,and these changes were closely related to the arginine biosynthesis pathway.Conclusions:The tumor-bearing mouse model with PD-1 monoclonal antibody resistance was successfully established.The changes in its peripheral serum metabolomics mainly involve arginine metabolism and the related changes of aspartate,malate and glutamate.

Programmed cell death plays an important role in the growth and development of organ-ism and homeostasis of tissues and organs.Necroptosis,a new mode of programmed cell death,characterized by necrosis in morphology,can cause cell breakdown and release of a significant num-ber of damage-associated molecular patterns,which play an important role in the occurrence and development of diseases.Viral infection is a serious health risk to both humans and domestic ani-mals.Research has confirmed that necroptosis functions as a cell death pathway in viral infections.In this paper,we review the current research advances on necroptosis related to viral infections,e-lucidate the molecular mechanism of mutual regulation between viral infection and necroptosis,and discuss the progress in the application of necroptosis inhibitors,to provide new ideas for the pre-vention and treatment of viral infections.

Objective:To investigate the predictive value of anti-Müllerian hormone (AMH) on the outcome of microscopic testicular sperm extraction (micro-TESE) in patients with non-mosaic Klinefelter syndrome (KS) of the clinical data and to identify effective predictors for successful micro-TESE.Methods:A retrospective case-control study was conducted on the clinical data of 118 non-mosaic KS patients treated at the Center for Reproductive Medicine of the First Affiliated Hospital of Zhengzhou University between May 2018 and September 2023. Patients were divided into two groups based on whether sperm were successfully retrieved via micro-TESE: the sperm retrieved group ( n=45) and the no sperm retrieved group ( n=73). Differences between the two groups were compared, and multivariate logistic regression analysis was used to identify factors influencing sperm retrieval. Changes in testicular volume and sex hormone levels before and after surgery were also assessed. Results:The sperm retrieval rate was 38.1% (45/118). Patients in the sperm retrieved group were significantly younger [(26.93±3.80) years] than those in the no sperm retrieved group [(28.27±3.92) years, P=0.029], and the AMH level was significantly higher [0.44 (0.18, 1.13) μg/L] than that in the no sperm retrieved group [0.10 (0.03, 0.22) μg/L, P<0.001]. AMH was identified as an independent predictor of micro-TESE outcome in non-mosaic KS patients ( OR=7.867, 95% CI: 2.727-27.242, P=0.001). The area under the receiver operating characteristic curve was 0.802 (95% CI: 0.722-0.883), and the optimal reference threshold for AMH was ≥0.265 μg/L. Postoperatively, testosterone levels decreased significantly by a median of 0.27 μg/L ( P=0.019), while luteinizing hormone levels increased by a median of 2.08 U/L ( P=0.049), with a more significant decline in testosterone levels observed in the no sperm retrieved group by a median of 0.29 μg/L ( P=0.022). Conclusion:AMH can predict successful micro-TESE in non-mosaic KS patients, with higher AMH levels indicating a higher likelihood of success.

Objective To construct mice hepatocellular carcinoma models with different tumor immune microenvironments(TIME)and explore the differences.Methods H22 and hepa1-6 were used to construct subcutaneous transplantation tumor model of C57 mice as homologous hepatocellular carcinoma cell lines(denoted as H22 group and hepal-6 group,each n=8),and the differences of TIME were evaluated.Immunohistochemistry was used to detect and quantify the infiltration of T cells,CD4+T cells,CD8+T cells,regulatory T cells and B cells in TIME.Flow cytometry was performed to detect the differences of composition of immune cell subpopulations in peripheral blood and tumor parenchyma.Gene expression profile characteristics of tumor tissue were analyzed based on high-throughput transcriptome sequencing technology,and enrichment analyses of immune-related signaling pathways were evaluated combined with gene ontology(GO)and Kyoto encyclopedia of genes and genomes(KEGG).Results H22 group showed cold and hepa1-6 group showed hot TIME characteristics.The number of T cells,CD4+T cells and CD8+T cells in tumor tissue of H22 group were all lower,while the proportion of T cells,CD4+T cells and CD8+T cells in peripheral blood were all higher than those of hepa1-6 group(all P＜0.05).Compared with H22 group,up-regulated genes of tumor tissue in hepa1-6 group expressed significantly enriched in tumor immune activation-related signaling pathways.Conclusion H22 and hepa1-6 hepatocellular carcinoma models showed distinct TIME characteristics of cold and hot tumors,respectively,and the amount of immune cells in tumor tissue of the former were significantly lower than those in the latter.

Objective:To investigate the influence of contrast-enhanced computed tomography(CECT)on dose calculation in magnetic resonance imaging(MRI)-guided online adaptive radiotherapy(oART)based on the electron density(ED)assignment method for rectal cancer.Methods:A retrospective analysis was conducted on the medical data of 15 patients with locally advanced rectal cancer at middle-low segments,who admitted to the Chinese PLA General Hospital between December 2023 and April 2025.All patients underwent both plain computed tomography(PCT)and CECT scans during location.The average HU and ED value of all organs that were extracted from PCT and CECT images in the treatment plan system were obtained,and the influences of contrast agent of intake on image characteristics of the structure of each organ(small intestine,femoral head,bladder)were analyzed.PCT was used as referred image to design reference plan(Pref).The synthetic CT(sCT)was simulated and generated on the basis of PCT and CECT,respectively.The beam flow field that was same with Pref was used to recalculate dose on sCT,and then,the online plan(PPCT)based on PCT,and the online plan(PCECT)based on CECT were obtained,respectively,which can simulate the online dose calculation of MRI-guided online adaptive radiotherapy(oART).The Pref was used as reference to compare dosimetric parameters for target region and organ at risk(OAR)through dose volume histogram(DVH)and planed evaluation indicators.Additionally,three dimension(3D)slicer software was used to perform γ analysis for the results of dose distribution,and explore the differences among PPCT,PCECT and Pref on dose distribution.Results:In terms of image characteristics,the HU values of soft-tissue organs(intestine,bladder,spinal cord,soft tissue)and planning target volume(PTV)in CECT were higher than those in PCT,and the differences of them were statistically significant(Zintestines=-2.188,Zbladder=-3.196,tspinal cord=-3.767,tsoft tissue=-10.083,tPTV=-4.693,P<0.05),while its influence was less on bone tissue.The statistical results of ED were consistent with those of HU.Regarding to dosimetric parameters,there was no statistically significant difference in target coverage rate between PPCT and Pref(P>0.05),and the D50%of the PPCT[(2724.25±19.91)cGy]was higher than that of the Pref[(2718.99±21.13)cGy],and the difference was statistically significant(t=-3.679,P=0.002).However,the target coverage rate of PCECT was 94.65(94.04,95.27)%,and the difference of that between PCECT and Pref was statistically significant(Z=-2.158,P=0.031).For OAR,the differences of Dmax value of the small intestine,and the V20 of the left femoral head between PPCT and Pref were significant(Z=-2.556,-2.529,P<0.05).The differences of the Dmax of small intestine,and the D50%of bladder between PCECT and Pref were significant(t=-4.821,2.171,P<0.05).The comparative γ passed rates of PPCT,PCECT and Pref under the standards of 3 mm/3%and 2 mm/2%were all above 95%,and the differences were not significant(P>0.05).Conclusions:The influence of CECT on dose calculation in MRI-guided oART based on ED assignment method for rectal cancer is relatively small,which can be directly used in the design of reference plan,but the maximum dose of radiation-sensitive organs such as the small intestine should be paid attention.

Objective:To explore the effects of Baduanjin and brisk walking on the sleep quality among fe-male college students.Methods:Ninety female college students with poor sleep quality[Pittsburgh Sleep Quality Index(PSQI)≥ 8]were recruited randomly assigned to Baduanjin,brisk walking,and control groups,with 30 par-ticipants in each.The Baduanjin and brisk walking groups participated in 10-week intervention(five 45-minute ses-sions per week),while the control group did not receive any intervention.Baseline and post-intervention assessments were conducted using the PSQI,a lung capacity test,echocardiography,and the Fatigue Scale(FS-14).Results:Af-ter 10 weeks,participants in both the Baduanjin and brisk walking groups got significantly lower PSQI and FS-14 total scores compared to baseline(Ps＜0.001).Cardiopulmonary function indicators,including stroke volume(SV),forced expiratory volume in one second(FEV1.0),the vital capacity-to-body mass index(VC/W),and maximum voluntary ventilation per minute(MVV),also significantly improved(Ps＜0.001).Furthermore,the Baduanjin group had significantly lower PSQI and FS-14 scores than both the brisk walking and control groups(P＜0.001),along with superior improvements in cardiopulmonary function(P＜0.001).Conclusion:This study in-dicates that Baduanjin is particularly effective in improving sleep quality,cardiopulmonary function,and reducing fatigue among female college students,showing advantages over brisk walking.

BACKGROUND:Allogeneic hematopoietic stem cell transplantation is an important treatment for malignant hematological diseases,and delayed postoperative platelet implantation is a common complication that seriously affects the quality of patient survival;however,there are no standard protocols to improve platelet implantation rates and prevent platelet implantation delays. OBJECTIVE:To compare the safety and efficacy of oral Herombopag Olamine versus subcutaneous recombinant human thrombopoietin for promoting platelet implantation in patients with malignant hematological diseases undergoing haploid hematopoietic stem cell transplantation. METHODS:Clinical data of 163 patients with malignant hematological diseases who underwent haploidentical hematopoietic stem cell transplantation from January 2016 to October 2022 were retrospectively analyzed.A total of 72 patients who started to subcutaneously inject recombinant human thrombopoietin at+2 days were categorized into the recombinant human thrombopoietin group;a total of 27 patients who started to orally take Herombopag Olamine at+2 days were categorized into the Herombopag Olamine group;and 64 patients who did not apply Herombopag Olamine or recombinant human thrombopoietin were categorized into the blank control group.The implantation status,incidence of acute graft-versus-host disease of degree II-IV within 100 days,1-year survival rate,1-year recurrence rate,and safety were analyzed in the three groups. RESULTS AND CONCLUSION:(1)The average follow-up time was 52(12-87)months.The implantation time of neutrophils in the blank control group,recombinant human thrombopoietin group,and Herombopag Olamine group was(12.95±3.88)days,(14.04±3.71)days,and(13.89±2.74)days,respectively,with no statistically significant difference(P=0.352);the implantation time of platelets was(15.16±6.27)days,(17.67±6.52)days,and(17.00±4.75)days,with no statistically significant difference(P=0.287).(2)The complete platelet implantation rate on day 60 was 64.06%,90.28%,and 92.59%,respectively,and the difference was statistically significant(P<0.001).The subgroup analysis showed that the difference between the blank control group and the recombinant human thrombopoietin group was statistically significant(P<0.001),and the difference between the blank control group and the Herombopag Olamine group was statistically significant(P=0.004).The difference was not statistically significant between the recombinant human thrombopoietin group and Herombopag Olamine group(P=0.535).(3)100-day II-IV degree acute graft-versus-host disease incidence in the blank control group,recombinant human thrombopoietin group,and Herombopag Olamine group were 25.00%,30.56%,and 25.93%,respectively,and the difference was not statistically significant(P=0.752).(4)The incidence of cytomegalovirus anemia,cytomegalovirus pneumonia,and hepatic function injury had no statistical difference among the three groups(P>0.05).(5)During the follow-up period,there was no thrombotic event in any of the three groups of patients.(6)The results showed that recombinant human thrombopoietin and Herombopag Olamine could improve the platelet implantation rate of malignant hematological disease patients after haploidentical hematopoietic stem cell transplantation,with comparable efficacy and good safety.

Programmed cell death plays an important role in the growth and development of organ-ism and homeostasis of tissues and organs.Necroptosis,a new mode of programmed cell death,characterized by necrosis in morphology,can cause cell breakdown and release of a significant num-ber of damage-associated molecular patterns,which play an important role in the occurrence and development of diseases.Viral infection is a serious health risk to both humans and domestic ani-mals.Research has confirmed that necroptosis functions as a cell death pathway in viral infections.In this paper,we review the current research advances on necroptosis related to viral infections,e-lucidate the molecular mechanism of mutual regulation between viral infection and necroptosis,and discuss the progress in the application of necroptosis inhibitors,to provide new ideas for the pre-vention and treatment of viral infections.