ObjectiveTo investigate the mechanism of action of remifentanil （RMZL） in alleviating lung ischemia-reperfusion injury （LIRI） in rats by inhibiting pyroptosis through modulating hypoxia inducible factor-1α （HIF-1α）/NOD-like receptor thermal protein domain associated protein 3 （NLRP3） pathway. MethodsRats were stochastically assigned into Control group， LIRI group， RMZL low-dose group， RMZL medium-dose group， RMZL high-dose group， and RMZL high-dose+HIF-1α activator dimethyloxallyl glycine （DMOG） group， with 18 rats in each group. Rats in Control group only had their left pulmonary hilum free and did not undergo ischemia-reperfusion treatment. Except for the Control group， LIRI models were constructed in all other groups. Rats in LIRI group were intraperitoneally injected with an equal amount of physiological saline 15 minutes before constructing LIRI model； rats in Control group were intraperitoneally injected with an equal amount of physiological saline 15 minutes before freeing left pulmonary hilum； rats in other groups were intraperitoneally injected with corresponding dose of drug 15 minutes before constructing LIRI model. The wet/dry weight ratio of lungs was calculated. HE staining was used to study lung tissue pathology. Immunofluorescence staining was used to detect the relative fluorescence intensity of gasdermin D （GSDMD） and NLRP3 double positive cells in lung tissue. ELISA was used to detect interleukin-1β and IL-18 in lung tissue. Western blot was used to detect HIF-1α， NLRP3， cysteine-aspartic protease-1 （Cleaved caspase-1）， and gasdermin D-N （GSDMD-N） proteins in lung tissue. ResultsCompared to the Control group， the LIRI group showed disordered alveolar structure， thickened alveolar septa， and abundant inflammatory cell infiltration in rats. The lung wet/dry weight ratio， relative fluorescence intensity of GSDMD and NLRP3 double positive cells in lung tissue， IL-1β， IL-18 levels， and HIF-1α， NLRP3， Cleaved caspase-1， and GSDMD-N proteins increased （P0.05）. For the LIRI group， rats in the RMZL low， medium， and high-dose groups displayed attenuated alveolar septal thickening and reduced inflammatory cell infiltration. The lung wet/dry weight ratio， relative fluorescence intensity of GSDMD and NLRP3 double positive cells in lung tissue， IL-1β， IL-18 levels， and HIF-1α， NLRP3， Cleaved caspase-1， and GSDMD-N proteins declined， and the RMZL high-dose group showed the most prominent trend （P0.05）. Compared with the RMZL high-dose group， rats in the RMZL high-dose+DMOG group exhibited thickened alveolar septa and more inflammatory cell infiltration， along with increased lung wet/dry weight ratio， relative fluorescence intensity of GSDMD and NLRP3 double positive cells in lung tissue， levels of IL-1β and IL-18， and protein expression of HIF-1α， NLRP3， Cleaved caspase-1， and GSDMD-N （P0.05）. ConclusionRMZL may inhibit pyroptosis in LIRI rats by suppressing HIF-1α/NLRP3 pathway.

The primary role of transfer RNA(tRNA)is to connect a specific amino acid to its 3' end, use its anticodon to match the codon on messenger RNA(mRNA), and deliver the corresponding amino acid to the ribosome for protein synthesis.tRNA exists in two forms: precursor tRNA and mature tRNA.When acted upon by enzymes like Dicer, elaC ribonuclease Z 2(ELAC2), angiopoietin(ANG), and other ribonucleases, tRNA is broken down into tRNA-derived stress-induced RNA(tiRNA)and tRNA-derived fragments(tRF).Recent advancements in RNA sequencing technology have led to increased interest in tiRNA and tRF, shedding light on their roles in various physiological and pathological processes.tRNA-derived small molecules(tsRNA)function similarly to microRNA(miRNA), influencing gene expression and protein synthesis.They show promise as diagnostic markers and potential therapeutic targets for age-related diseases.This review offers a comprehensive analysis of tsRNA classification, biological functions, research advancements, and clinical applications in age-related conditions.

Ursodeoxycholic acid (UDCA) is a naturally occurring, low-toxicity, and hydrophilic bile acid (BA) in the human body that is converted by intestinal flora using primary BA. Solute carrier family 7 member 11 (SLC7A11) functions to uptake extracellular cystine in exchange for glutamate, and is highly expressed in a variety of human cancers. Retroperitoneal liposarcoma (RLPS) refers to liposarcoma originating from the retroperitoneal area. Lipidomics analysis revealed that UDCA was one of the most significantly downregulated metabolites in sera of RLPS patients compared with healthy subjects. The augmentation of UDCA concentration (≥25 μg/mL) demonstrated a suppressive effect on the proliferation of liposarcoma cells. [¹⁵N₂]-cystine and [¹³C₅]-glutamine isotope tracing revealed that UDCA impairs cystine uptake and glutathione (GSH) synthesis. Mechanistically, UDCA binds to the cystine transporter SLC7A11 to inhibit cystine uptake and impair GSH de novo synthesis, leading to reactive oxygen species (ROS) accumulation and mitochondrial oxidative damage. Furthermore, UDCA can promote the anti-cancer effects of ferroptosis inducers (Erastin, RSL3), the murine double minute 2 (MDM2) inhibitors (Nutlin 3a, RG7112), cyclin dependent kinase 4 (CDK4) inhibitor (Abemaciclib), and glutaminase inhibitor (CB839). Together, UDCA functions as a cystine exchange factor that binds to SLC7A11 for antitumor activity, and SLC7A11 is not only a new transporter for BA but also a clinically applicable target for UDCA. More importantly, in combination with other antitumor chemotherapy or physiotherapy treatments, UDCA may provide effective and promising treatment strategies for RLPS or other types of tumors in a ROS-dependent manner.

BACKGROUND:Previously,a SENP1 gene-silenced human alveolar epithelial cell line was successfully constructed in vitro,and the role of SENP1 in hyperoxic lung injury was investigated at the cellular level.OBJECTIVE:To construct a mouse model of alveolar type II epithelial cell-specific knockout of SENP1 gene based on the Cre-loxP recombinase system.METHODS:SENP1flox/-mice were self-crossed to obtain SENP1flox/flox and SENP1flox/-mice;Sftpc-Cre+/+mice were crossed with wild-type mice to obtain more Sftpc-Cre+/-mice.Sftpc-Cre+/+or offspring Sftpc-Cre+/-mice were crossed with SENP1flox/-or offspring SENP1flox/flox mice to obtain SENP1flox/-Sftpc-Cre+/-double heterozygous mice.SENP1flox/-Sftpc-Cre+/-mice were then crossed with SENP1flox/flox mice to obtain SENP1flox/floxSftpc-Cre+/-mice.The genomic DNA was extracted by tail clipping and amplified by PCR.The amplified product was subjected to agarose gel electrophoresis to determine the mouse genotypes.Lung tissues of SENP1flox/flox and SENP1flox/floxSftpc-Cre+/-mice were subjected to immunofluorescence double-labelling and western blot assay to verify the knockdown effect of SENP1 gene.Heart,liver,lung and kidney tissues of SENP1flox/flox and SENP1flox/floxSftpc-Cre+/-mice were stained with hematoxylin-eosin to observe the histomorphology of each organ in the two groups of mice.RESULTS AND CONCLUSION:SENP1flox/floxSftpc-Cre+/-mice were correctly screened by agarose gel electrophoresis.Immunofluorescence double-labeling experiments showed that the mean fluorescence intensity of SENP1 was reduced in lung tissues of SENP1flox/floxSftpc-Cre+/-mice compared with that of SENP1flox/flox mice(P<0.01)and no significant co-localization of SENP1 and Sftpc was observed(P<0.01).Western blot results showed that SENP1 protein expression was reduced in lung tissues of SENP1flox/floxSftpc-Cre+/-mice compared with SENP1flox/flox mice(P<0.001).Hematoxylin-eosin staining showed no significant alterations in the histomorphology of heart,liver,lung and kidney tissues in SENP1flox/flox and SENP1flox/floxSftpc-Cre+/-mice.This study successfully constructed alveolar type II epithelial cell-specific knockout SENP1 gene mice using the Cre-loxP recombinase system,which provides a good tool for the subsequent study of the role of SENP1 gene in lung diseases such as bronchopulmonary dysplasia and idiopathic pulmonary fibrosis,in which alveolar type II epithelial cells are the main damage cells.

Ursodeoxycholic acid(UDCA)is a naturally occurring,low-toxicity,and hydrophilic bile acid(BA)in the human body that is converted by intestinal flora using primary BA.Solute carrier family 7 member 11(SLC7A11)functions to uptake extracellular cystine in exchange for glutamate,and is highly expressed in a variety of human cancers.Retroperitoneal liposarcoma(RLPS)refers to liposarcoma originating from the retroperitoneal area.Lipidomics analysis revealed that UDCA was one of the most significantly down-regulated metabolites in sera of RIPS patients compared with healthy subjects.The augmentation of UDCA concentration(≥25 μg/mL)demonstrated a suppressive effect on the proliferation of liposarcoma cells.[15N2]-cystine and[13Cs]-glutamine isotope tracing revealed that UDCA impairs cystine uptake and glutathione(GSH)synthesis.Mechanistically,UDCA binds to the cystine transporter SLC7A11 to inhibit cystine uptake and impair GSH de novo synthesis,leading to reactive oxygen species(ROS)accumulation and mitochondrial oxidative damage.Furthermore,UDCA can promote the anti-cancer effects of ferroptosis inducers(Erastin,RSL3),the murine double minute 2(MDM2)inhibitors(Nutlin 3a,RG7112),cyclin dependent kinase 4(CDK4)inhibitor(Abemaciclib),and glutaminase inhibitor(CB839).Together,UDCA functions as a cystine exchange factor that binds to SLC7A11 for antitumor activity,and SLC7A11 is not only a new transporter for BA but also a clinically applicable target for UDCA.More importantly,in combination with other antitumor chemotherapy or physiotherapy treatments,UDCA may provide effective and promising treatment strategies for RLPS or other types of tumors in a ROS-dependent manner.

In recent years,adoptive T cell immunotherapy has become a research hotspot in cancer treat-ment,in which the directional homing of T cells to tumor tissues is the core of the anti-tumor immune re-sponse,which is closely related to good clinical treatment outcomes.However,the infiltration of T cells into solid tumors remains challenging due to the complex tumor microenvironment,tumor vasculature bar-riers,and loss of chemotaxis signals.This review systematically outlines the migration pathways of T cell homing,including blood homing after intravenous infusion,transvascular endothelial migration,and its infiltration into targeted solid tumor tissues.On this basis,we also explore the regulatory mechanism of T cell homing,especially the synergistic relationship between the chemokine-receptor axis,and the effects of tumor vascular abnormalities,tumor microenvironment(TME)-shaped infiltration barriers,and tumor stromal barriers on T cell homing.In response to the above obstacles,three main strategies to enhance the homing efficiency of T cells are reviewed.First,chemokine receptors(e.g.,CXCR2,CXCR6)are modified to match tumor chemotaxis signals,or immune checkpoint molecules(PD-1,LAG-3,SHP-1)are knocked out to reverse T cell exhaustion by CRISPR gene editing or lentiviral transduction technolo-gy.Second,targeting the VEGF/VEGFR axis combined with ATCT can promote vascular normalization and improve T cell infiltration.Third,the combination of local therapy(radiotherapy,oncolytic virus)or systemic drugs(chemotherapy,immune checkpoint inhibitors,etc.)can improve the homing of T cells by remodeling tumor TME.These strategies will provide a theoretical basis and research direction for a-doptive T cell immunotherapy in the treatment of solid tumors.

[Purpose]To analyze the trends of cancer incidence and mortality from 1991 to 2020 in Jiashan County of Zhejiang Province,and to predict trends from 2021 to 2035.[Methods]Cancer cases and deaths from 1991 to 2020 in Jiashan County were collected.The crude inci-dence rate and mortality rate,age-standardized rate(ASR)were calculated.The annual percentage change(APC)and average annual percentage change(AAPC)were calculated by Joinpoint model to analyze the trend of cancer incidence and mortality.The age-period-cohort model was used to predict the trends of cancer incidence and mortality from 2021 to 2035 and the estimated annual percentage change(EAPC)was calculated.[Results]The age-standardized incidence rates by Chinese standard population(ASIRC)in Jiashan were 159.06/105,187.25/105,and 254.37/105 during 1991-2000,2001-2010,and 2011-2020,respectively.The trends in young and middle-aged women and people above 65 years old were more predominant.Lung,stomach,colorectal,liver and female breast cancers were major incident cancers for the period from 1991 to 2020.The cancer incidence showed an increasing trend from 1991 to 2020(AAPC=2.17％,P＜0.05)and is expect-ed to continue to increase over 2021 to 2035(EAPC=3.03％,P＜0.05).The age-standardized mor-tality rates by Chinese standard population(ASMRC)in Jiashan were 119.74/105,117.79/105,and 100.11/105 during 1991-2000,2001-2010,and 2011-2020,respectively.Lung,liver,stomach,colorectal,esophageal,and pancreatic cancers were leading causes of cancer deaths from 1991 to 2020.The cancer mortality showed a decreasing trend from 1991 to 2020(AAPC=-1.00％,P＜0.05)and is expected to continue to decline over 2021 to 2035(EAPC=-1.67％,P＜0.05).[Conclusion]The mortality rate from cancers has been gradually declined in Jiashan County,while the incidence rate has increased significantly and will continue to increase in the next 15 years.High incidence of lung cancer,liver cancer,gastric cancer and colorectal cancer makes it crucial to pay close attention to key groups and strengthen the control of risk factors of high inci-dence cancer.

In recent years,adoptive T cell immunotherapy has become a research hotspot in cancer treat-ment,in which the directional homing of T cells to tumor tissues is the core of the anti-tumor immune re-sponse,which is closely related to good clinical treatment outcomes.However,the infiltration of T cells into solid tumors remains challenging due to the complex tumor microenvironment,tumor vasculature bar-riers,and loss of chemotaxis signals.This review systematically outlines the migration pathways of T cell homing,including blood homing after intravenous infusion,transvascular endothelial migration,and its infiltration into targeted solid tumor tissues.On this basis,we also explore the regulatory mechanism of T cell homing,especially the synergistic relationship between the chemokine-receptor axis,and the effects of tumor vascular abnormalities,tumor microenvironment(TME)-shaped infiltration barriers,and tumor stromal barriers on T cell homing.In response to the above obstacles,three main strategies to enhance the homing efficiency of T cells are reviewed.First,chemokine receptors(e.g.,CXCR2,CXCR6)are modified to match tumor chemotaxis signals,or immune checkpoint molecules(PD-1,LAG-3,SHP-1)are knocked out to reverse T cell exhaustion by CRISPR gene editing or lentiviral transduction technolo-gy.Second,targeting the VEGF/VEGFR axis combined with ATCT can promote vascular normalization and improve T cell infiltration.Third,the combination of local therapy(radiotherapy,oncolytic virus)or systemic drugs(chemotherapy,immune checkpoint inhibitors,etc.)can improve the homing of T cells by remodeling tumor TME.These strategies will provide a theoretical basis and research direction for a-doptive T cell immunotherapy in the treatment of solid tumors.

BACKGROUND:Previously,a SENP1 gene-silenced human alveolar epithelial cell line was successfully constructed in vitro,and the role of SENP1 in hyperoxic lung injury was investigated at the cellular level.OBJECTIVE:To construct a mouse model of alveolar type II epithelial cell-specific knockout of SENP1 gene based on the Cre-loxP recombinase system.METHODS:SENP1flox/-mice were self-crossed to obtain SENP1flox/flox and SENP1flox/-mice;Sftpc-Cre+/+mice were crossed with wild-type mice to obtain more Sftpc-Cre+/-mice.Sftpc-Cre+/+or offspring Sftpc-Cre+/-mice were crossed with SENP1flox/-or offspring SENP1flox/flox mice to obtain SENP1flox/-Sftpc-Cre+/-double heterozygous mice.SENP1flox/-Sftpc-Cre+/-mice were then crossed with SENP1flox/flox mice to obtain SENP1flox/floxSftpc-Cre+/-mice.The genomic DNA was extracted by tail clipping and amplified by PCR.The amplified product was subjected to agarose gel electrophoresis to determine the mouse genotypes.Lung tissues of SENP1flox/flox and SENP1flox/floxSftpc-Cre+/-mice were subjected to immunofluorescence double-labelling and western blot assay to verify the knockdown effect of SENP1 gene.Heart,liver,lung and kidney tissues of SENP1flox/flox and SENP1flox/floxSftpc-Cre+/-mice were stained with hematoxylin-eosin to observe the histomorphology of each organ in the two groups of mice.RESULTS AND CONCLUSION:SENP1flox/floxSftpc-Cre+/-mice were correctly screened by agarose gel electrophoresis.Immunofluorescence double-labeling experiments showed that the mean fluorescence intensity of SENP1 was reduced in lung tissues of SENP1flox/floxSftpc-Cre+/-mice compared with that of SENP1flox/flox mice(P<0.01)and no significant co-localization of SENP1 and Sftpc was observed(P<0.01).Western blot results showed that SENP1 protein expression was reduced in lung tissues of SENP1flox/floxSftpc-Cre+/-mice compared with SENP1flox/flox mice(P<0.001).Hematoxylin-eosin staining showed no significant alterations in the histomorphology of heart,liver,lung and kidney tissues in SENP1flox/flox and SENP1flox/floxSftpc-Cre+/-mice.This study successfully constructed alveolar type II epithelial cell-specific knockout SENP1 gene mice using the Cre-loxP recombinase system,which provides a good tool for the subsequent study of the role of SENP1 gene in lung diseases such as bronchopulmonary dysplasia and idiopathic pulmonary fibrosis,in which alveolar type II epithelial cells are the main damage cells.

The primary role of transfer RNA(tRNA)is to connect a specific amino acid to its 3' end, use its anticodon to match the codon on messenger RNA(mRNA), and deliver the corresponding amino acid to the ribosome for protein synthesis.tRNA exists in two forms: precursor tRNA and mature tRNA.When acted upon by enzymes like Dicer, elaC ribonuclease Z 2(ELAC2), angiopoietin(ANG), and other ribonucleases, tRNA is broken down into tRNA-derived stress-induced RNA(tiRNA)and tRNA-derived fragments(tRF).Recent advancements in RNA sequencing technology have led to increased interest in tiRNA and tRF, shedding light on their roles in various physiological and pathological processes.tRNA-derived small molecules(tsRNA)function similarly to microRNA(miRNA), influencing gene expression and protein synthesis.They show promise as diagnostic markers and potential therapeutic targets for age-related diseases.This review offers a comprehensive analysis of tsRNA classification, biological functions, research advancements, and clinical applications in age-related conditions.