abstract:
Objective To construct Csf1rCreRosa26YFPmice,and to detect the efficiency of Csf1rCre-mediated yellow fluorescent protein(YFP) labeling monocytes and tissue-resident macrophages in different tissues. Methods The Rosa26YFPmutant mice had a loxP-flanked STOP sequence,followed by a yellow fluorescent protein gene(YFP)inserted into the Rosa26 locus. When crossed with mice expressing the Csf1rCrerecombinase,the STOP sequence was deleted,and yellow fluorescent protein(YFP) expression was observed in the tissues of double-mutant offspring(Csf1rCreRosa26YFP). Csf1rCremice were mated with Rosa26YFPmice,and genotype of Csf1rCreRosa26YFPmice were identified by PCR. Blood and bone marrow monocytes,liver macrophages,kidney macrophages,alveolar macrophages and spleen macrophages of adult Csf1rCreRosa26YFPmice were isolated,labeled with flow cytometry antibodies,and the recombination efficiency of Csf1rCre-mediated YFP labeling tissue macrophages was analyzed by flow cytometry. Results In Csf1rCreRosa26YFPreporter mice,the median percentage of YFP + was 93. 25% in renal macrophages,92. 45% in liver macrophages,and 91. 10% in spleen macrophages. The percentage of YFP +was 94. 70% in alveolar macrophages,98. 20% in blood monocytes,and 93. 90% in bone marrow monocytes.Conclusion Csf1rCrecan be used to trace tissue-resident macrophages as well as bone marrow and blood monocytes. At the same time,Csf1rCrecan target these cells to prepare conditional gene knockout mice.

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To investigate the mechanism by which icariin ( ICA) inhibits the invasion and metastasis of human triple-negative breast cancer ( TNBC) cells via downregulation of the transforming growth factor-β/ Smad ( TGF-β/ Smad) signaling pathway．Methods TNBC cells ( MDA-MB-231 and MDA-MB-468) were cultured in vitro and divided into four groups: an experimental group treated with 15 μmol / L ICA; a model group treated with 10 μmol / L TGF-β receptor inhibitor LY2109761; a combination group ( LY2109761 + ICA) treated with both 15 μmol / L ICA and 10 μmol / L LY2109761; and a control group．Cell proliferation，migration，and invasion were as- sessed using CCK-8，colony formation，5-ethynyl-2 '-deoxyuridine ( EdU) ，wound healing，and Transwell assays． The expression levels of epithelial-mesenchymal transition ( EMT) -related proteins，as well as TGF-β1，Smad2， and phosphorylated Smad2 ( P-Smad2) were detected by immunofluorescence and Western blot．Results CCK-8 results showed that cell proliferation decreased gradually with increasing concentrations of ICA ( P＜0. 05) ．Colony formation and EdU assays indicated significantly inhibited proliferation in the ICA-treated group compared to the control ( P＜0. 05) ．Wound healing and Transwell assays demonstrated reduced migration and invasion capabilities in the experimental group relative to the control ( P＜0. 05) ．Compared to the model group，the LY2109761 + ICA group exhibited further suppression of invasion ( P＜0. 05) ．Immunofluorescence revealed decreased Vimentin ex- pression in the experimental group ( P＜0. 05) ，with an even more pronounced reduction in the LY2109761 + ICA group ( P＜0. 01) ．Western blot analysis showed that the protein levels of N-cadherin，matrix metalloproteinase-9( MMP9) ，Vimentin，TGF-β1，Smad2，and P-Smad2 were downregulated in the experimental group compared to the control ( P＜0. 05) ．These proteins were further suppressed in the LY2109761 + ICA group compared to the model group ( P＜0. 05) ．Conclusion ICA inhibits TNBC cells proliferation，invasion，metastasis，and EMT by downregulating the TGF-β/ Smad signaling pathway．

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Objective To perform transient transfection of recombinant human Factor X(rhFX) into HEK293 cells,to optimize transfection parameters,to develop a high-yield in vitro expression system for rhFX production,and to assess the biological activity of expressed rhFX. Methods The eukaryotic expression vector pcDNA3. 1-EGFP-FX was constructed by inserting the F10 gene into pcDNA3. 1-EGFP and subsequently transfected into HEK293cells to validate the transfection system efficiency. The recombinant expression vector pcDNA3. 1-FX was generated through ligation of the F10 gene fragment with pcDNA3. 1, followed by liposome-mediated transfection into HEK293 cells. The expression of rhFX in the cell supernatant was analyzed by Western blot and sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE). Transfection conditions were systematically optimized,and rhFX concentration was quantified by enzyme-linked immunosorbent assay(ELISA). The coagulation bioactivity of rhFX was evaluated through prothrombin time(PT) assay and chromogenic substrate method. Results rhFX was successfully expressed in the supernatant of HEK293 cells. rhFX was successfully expressed in the supernatant of HEK293 cells. The highest expression level of rhFX was achieved on the third day after transfection when the cell density was 80% and the ratio of plasmid DNA to polyethyleneimine(PEI) transfection reagent was 1 ∶ 2.Triplicate ELISA measurements demonstrated a maximum rhFX concentration of 5. 20 ng/mL in the supernatant.The prothrombin time(PT) of rhFX-containing supernatant was significantly shorter(P 50) of etoxaban was determined to be 1. 449 nmol/L using the chromogenic substrate method based on rhFX,which was in the same order of magnitude as the reported 0. 78 nmol/L in the literature. Conclusion HEK293cells successfully express biologically active recombinant human Factor X(rhFX) protein,laying a foundation for advancing the development of rhFX-based therapeutics.

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To establish and identify hepatocyte-specific transmembrane protein 121 ( Tmem121 ) knockout mice．Methods The hepatocyte-specific Tmem121 knockout mice ( Tmem121flox / flox / Cre，Tmem121ΔHep) were obtained by crossbreeding of Tmem121flox / + / Cre and Tmem121flox / flox mice，which were generated using the CRISPR / Cas9 and Cre / Loxp systems．The genotype was verified by PCR using genomic DNA extracted from mouse tails as template．The growth，reproduction and organ development of both control and knockout mice were ob- served and analyzed．PCR and Western blot methods were performed to assess the knockout efficiency of Tmem121 in mouse primary hepatocytes．CellMaskTM Deep Red plasma membrane staining was employed to compare the mor- phological differences in primary hepatocytes between control and knockout mice．Results Tmem121flox / flox / Cre mice were successfully obtained according to genotype identification analysis，and there were no significant differ- ences between control and knockout mice in body mass，reproductive ability，growth and development of liver．The specific knockout of Tmem121 gene in primary hepatocytes did not significantly affect the morphological structure or pathological characteristics of liver tissue．However，compared to the control group，the levels of Tmem121 mRNA and protein in the primary hepatocytes of the knockout group were significantly reduced ( P ＜0. 01) ．CellMaskTM Deep Red plasma membrane staining indicated that the proportion of binucleated hepatocytes in Tmem121-deficient mice significantly increased ( P＜0. 05) ，while the cell area was significantly reduced ( P＜0. 001) ．Conclusion Hepatocyte-specific Tmem121 knockout mice are successfully constructed，which provides an animal model for further exploration of the function and mechanism of Tmem121 gene in liver diseases．

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To investigate the therapeutic effect of bone marrow mesenchymal stem cells ( BMSCs) mod- ified by bone morphogenetic protein ( BMP) -2 gene combined with nano-hydroxyapatite ( nHA) / polyamide 66 ( PA66) on femoral nonunion in rats．Methods Rat BMSCs were isolated and divided into the stent + BMSCs group，stent + BMSCs + rhBMP-2 group，and stent + BMSCs + Ad-BMP-2 group; human recombinant BMP-2 ( rh- BMP-2) or adenovirus-infected BMP-2 ( Ad-BMP-2) vector was transfected into BMSCs and loaded onto nHA / PA66 stent materials．Flow cytometry was used to detect that BMSCs expressed specific surface markers．Cell pro- liferation was detected by MTT assay，related bioactive factors［platelet-derived growth factor ( PDGF) ，transfor- ming growth factor-β ( TGF-β) ，vascular endothelial growth factor ( VEGF) ，fibroblast growth factor ( FGF) ，os- teocalcin ( OCN) ，osteonectin ( ON) ］were detected by ELISA，alkaline phosphatase ( ALP) activity was detec- ted，and cell growth and adhesion were observed by scanning electron microscopy ( SEM) ．In the atrophic nonun- ion model of SD rats，the stent + BMSCs + rhBMP-2 or stent + BMSCs + Ad-BMP-2 complex was implanted into the defect area，which was divided into the rhBMP-2 group and the Ad-BMP-2 group，and the therapeutic effect was e- valuated by X-ray and MicroCT．Results The surface of the nHA / PA66 stent was smooth and porous，and BMSCs adhered well．Flow cytometry showed high expression of CD29 and CD90 and low expression of CD45 and CD34 in BMSCs．MTT showed that the cells proliferated rapidly after 72 h．Compared with the stent + BMSCs group and the stent + BMSCs + rhBMP-2 group，the ALP activity，the expressions of PDGF，TGF-β , VEGF，FGF，OCN，and ON in the stent + BMSCs + Ad-BMP-2 group were significantly up-regulated ( P＜0. 05) ．12 weeks after surgery， the stent complex in the Ad-BMP-2 group of rats was completely wrapped by newly formed cortical bone，and the surface was smooth and intact．X-ray showed that the complex in the Ad-BMP-2 group was completely wrapped by newly formed cortical bone，and the recovery degree was better than that in the rhBMP-2 group．Micro-CT results showed that compared with the rhBMP-2 group，the bone volume fraction，trabecular thickness，trabecular num- ber，bone mineral density，and bone mineral content in the Ad-BMP-2 group all increased 12 weeks after surgery ( P＜0. 05) ，and the trabecular separation level decreased ( P＜0. 05) ．Conclusion Ad-BMP-2-transfected BM- SCs combined with nHA / PA66 stent material can express bioactivity more effectively，provide a continuous and good microenvironment for the proliferation and differentiation of BMSCs，which is beneficial to promoting local os- teogenic activity and bone defect repair．

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Objective To investigate the impacts of remifentanil on the proliferation,apoptosis and Wnt/β-catenin pathway of breast cancer MDA-MB-231 cells. Methods Different concentrations of remifentanil(0,0. 5,2. 5,5. 0,10. 0,20. 0,40. 0 μg/mL) were used to treat human normal breast MCF-10A cells and breast cancer MDAMB-231 cells to screen for experimental concentrations of remifentanil. MDA-MB-231 cells were divided into control group,remifentanil low(5. 0 μg/mL),medium(10. 0 μg/mL),and high(20. 0 μg/mL) concentration groups,and remifentanil + SKL2001(Wnt/β-catenin pathway agonist) group. MTT assay and colony formation assay were applied to detect cell proliferation ability. Flow cytometry was applied to detect cell apoptosis and cell cycle changes. Western blot was applied to detect protein expression related to cell proliferation,apoptosis,and the Wnt/β-catenin signaling pathway. Results Remifentanil at concentrations of 5. 0,10. 0,and 20. 0 μg/mL could reduce the viability of MDA-MB-231 cells and had no prominent toxicity to MCF-10A cells. Compared with the control group,the optical density value of cell proliferation,colony formation number,proportion of S-phase cells,and the protein levels of cellular myelocytomatosis(c-Myc),Cyclin D1,B lymphoblastoma 2(Bcl-2),precursor of cysteine aspartate protease-3(pro-caspase-3) and β-catenin were lower in the low,medium,and high concentration remifentanil groups(P 0/G1 phase cells,apoptosis rate,early apoptosis rate,the protein levels of Bcl-2 associated X protein(Bax) and cleaved cysteine aspartate protease-3(Cleaved caspase-3),and glycogen synthase kinase-3β(GSK-3β) were higher( P ＜0. 05) ． SKL2001 could weaken the effects of remifentanil on the proliferation and apoptosis of MDA-MB-231 cells．Conclusion Remifentanil inhibits the proliferation of MDA-MB-231 cells，induces apoptosis and cell cycle arrest，and its mechanism may be related to the inhibition of Wnt / β-catenin signaling pathway activation．

abstract:
Objective To explore the mechanism of hesperidin on hindlimb motor function in mice with spinal cord injury(SCI). Methods Oxygen glucose deprivation(OGD) was used to induce SCI in PC12 neuronal cells.CCK-8 assay determined the optimal hesperidin concentration(0,10,20,30,40,50,60 mmol/L) for subsequent experiments after 24 h treatment. PC12 cells were divided into Control group,OGD group and Hesperidin(30 mmol/L) + OGD groups. Hoechst staining assessed apoptosis; Western blot(WB),qPCR and immunofluorescence(IF) detected the expression of i NOS,CD206 and TNF-α. Flow cytometry measured apoptosis rates,and WB examined the impact of hesperidin on PI3K,p-PI3K,AKT,p-AKT,and NF-κB expression. Forty-five healthy male C57BL/6 mice were randomly divided into Sham group,Sci group and Hesperidin groups. Sham and Sci groups received 0. 1 mL PBS,while the Hesperidin group received 0. 1 mL hesperidin. After modeling,i NOS,CD206,and TNF-α expression levels,spinal cord cavitation,and hindlimb motor function were evaluated. Results Hesperidin at 10-30 mmol/L for 24 h did not significantly affect PC12 cell activity. Hoechst staining showed reduced apoptosis in PC12 cells after 24 h of 30 mmol/L hesperidin treatment. Compared to the OGD group,Hesperidin + OGD group had decreased mRNA and protein expression of inflammatory factors(i NOS and TNF-α) and increased expression of the anti-in ammatory factor(CD206). Hesperidin treatment inhibited PI3K/AKT/NF-κB pathway activation,with similar results in mouse experiments. Compared to Sci group,Hesperidin group had reduced spinal cord cavitation area and improved hindlimb motor function. Conclusion Hesperidin may improve hindlimb motor function in mice with SCI by downregulating the phosphorylation level of the PI3K/AKT/NF-κB pathway,inhibiting in ammatory factor release,promoting anti-inflammatory factor release,regulating the inflammatory microenvironment after SCI,and suppressing the acute inflammatory response.

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Objective To investigate the causes of false negative results in the detection of Streptococcus agalactiae using fluorescent quantitative PCR(qPCR) targeting the CAMP factor gene(cfb),and to perform a comprehensive analysis of the associated molecular mechanisms. Methods A total of 76 vaginal secretion samples were evaluated using both qPCR based on cfb gene and bacterial culture methods. Four suspicious strains exhibiting negative qPCR results but positive culture findings were identified using matrix-assisted laser desorption ionization time-of-flight mass spectrometry(MALDI-TOF MS),latex agglutination antigen detection,and the CAMP test. Whole genome analysis was conducted utilizing the MGI DNBSEQ-T7 and Nanopore-PromethION 48 sequencing platforms. Phylogenetic and circular evolutionary trees were constructed using the 16S rRNA gene for strain verification. Multilocus sequence typing(MLST) was conducted,and cfb sequences were aligned and analyzed based on spliced sequences and original data. Specific primers targeting the cfb gene were designed for full-length amplification,followed by verification through agarose gel electrophoresis. Results The four strains identified as suspicious were classified as S. agalactiae through MALDI-TOF MS,antigen detection,and 16S rRNA gene analysis,with MLST typing indicating ST-862. Phenotypic analysis revealed a negative CAMP test. Whole genome sequence alignment failed to detect the cfb gene or any homologous sequences,and molecular testing confirmed the absence of cfb gene PCR amplification products,thereby confirming its complete deletion. Conclusion This deletion is identified as the molecular mechanism responsible for the false negative qPCR detection of S. agalactiae when targeting this specific gene.It is recommended that the qPCR detection targeting a single cfb gene has limitation,and this may impact clinical diagnosis and treatment decisions. This limitation warrants carefulconsideration.

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Objective To isolate a lytic phage targeting K. pneumoniae type K39 from untreated sewage and systematically analyze its biological characteristics and genomic information. Methods The bacteriophage of K. pneumoniae K39 Kp 1000 was identified based on the sequence of the capsular polysaccharide gene. It was isolated and purified using the double agar plate method. The morphology of the phage was observed through negative staining and transmission electron microscopy,and its bacteriophage spectrum was evaluated by phagocytosis. The biological characteristics of the bacteriophage were assessed by determining the optimal multiplicity of infection(MOI),constructing a one-step growth curve,and conducting temperature and pH tolerance tests. Finally,the phage DNA was extracted,and its whole genome was sequenced using the Illumina Hi Seq2000 sequencing platform. The sequencing results were then annotated and analyzed. Results A lytic phage specifically targeting K. pneumoniae type K39was successfully isolated and named IME330. Transmission electron microscopy showed that the head diameter of the phage was(75 ± 1) nm and the tail length was(185 ± 1) nm. The lytic spectrum analysis revealed that IME330 lysed K. pneumoniae type K39,demonstrating a narrow host range and high specificity. The optimal multiplicity of infection(MOI) was 0. 1,The lysis amount reached 168 PFU/cell. Physicochemical experiments indicated that IME330 exhibited strong tolerance to high temperatures and a broad p H range(pH 4-10). Genomic analysis demonstrated that the IME330 genome had a length of 144,245 bp,a molecular weight of 46,463 MDa,G +C content of 44. 8%,and encoded 320 open reading frames(ORFs). Conclusion IME330 is a novel lytic K.pneumoniae phage belonging to the order Caudovirales. This phage exhibits strong tolerance to physicochemical factors(e. g.,temperature,acidic/alkaline conditions) and demonstrates high lytic activity,while its lytic spectrum remains narrow with strict host specificity.

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Objective To investigate the ameliorative effect of vitamin D on sciatic nerve chronic compression injury(CCI) rats and its regulatory effect on nuclear factor κB(NF-κB)/cystathione-β-synthase(CBS)-hydrogen sulfide(H2S) system. Methods Fifty rats were randomly divided into five groups: control group,model group,vitamin D group,activator group,and activator + vitamin D group,with 10 rats in each group. Except for the control group,the rats in the other groups underwent sciatic nerve ligation to establish the CCI model. The vitamin D group received intraperitoneal injection of 500 mg/kg vitamin D,the activator group received intraperitoneal injection of10 mg/kg NF-κB activator lipopolysaccharide,and the activator + vitamin D group received intraperitoneal injection of 500 mg/kg vitamin D along with 10 mg/kg lipopolysaccharide. The control and model groups received intraperitoneal injection of an equal volume of saline,once per day for 2 weeks. The following parameters were compared among the groups: paw withdraw thermal latency(PWTL),mechanical withdrawal threshold(MWT),serum 25-hydroxyvitamin D3[25(OH)D3]levels,H2S content,calcitonin gene-related peptide(CGRP),prostaglandin E2(PGE2) levels,and the expression of vitamin D receptor(VDR),NF-κBp65,and CBS proteins in the L4-L6 dorsal root ganglia of rats. Results Compared with the control group,the model group showed decreased PWTL,MWT,25(OH)D3 levels and VDR protein expression,while H2S content,CGRP and PGE2 levels,and NF-κBp65 and CBS protein expression significantly increased(P 3 levels and VDR protein expression(P 2S content,CGRP and PGE2 levels,and NF-κBp65 and CBS protein expression significantly decreased(P3 levels and VDR protein expression,and decreased H2S content,CGRP and PGE2 levels and decreased NF-κBp65,CBS protein expression(P 3 levels and VDR protein expression,while H2S content,CGRP and PGE2 levels,and NF-κBp65 and CBS protein expression significantly increased(P 2S signaling pathway.

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To explore the effects and mechanisms of Kobophenol A ( KPA) on lipopolysaccharide ( LPS) -induced M1 macrophage polarization，and to provide a theoretical basis for the treatment of inflammatory immune diseases and the development of new drugs． Methods The M1 macrophage polarization model of RAW264. 7 was established by LPS induction，and the peroxiredoxin 6 ( Prdx6) knockdown model was constructed using the Prdx6 inhibitor MJ33 and Prdx6-siRNA．RAW264. 7 cells，a mouse macrophage cell line，were treated with various concentrations of KPA． Cell viability was assessed using the CCK-8 assay．The expression levels of Prdx6 and M1 macrophage polarization-related proteins，including inducible nitric oxide synthase ( iNOS) and cy- clooxygenase-2 ( COX-2) ，were detected by Western blot and immunofluorescence staining．The expression levels of Prdx6 and M1 macrophage polarization-related genes iNOS，interleukin-6 ( IL-6) ，and tumor necrosis factor α ( TNF-α) ，were measured by RT-qPCR. Flow cytometry was employed to detect the expression of cluster of differ- entiation 86 ( CD86) ，a marker of M1 macrophages．Results Compared with the LPS-induced M1 macrophage po- larization model ， KPA significantly reversed the morphological changes of M1 macrophage polarization in RAW264. 7 macrophages and decreased the expression of M1 macrophage polarization-related proteins iNOS，COX- 2，CD86 and related genes iNOS，IL-6，TNF-α ( all P＜0. 05) ．In addition，LPS significantly downregulated the expression of Prdx6 in RAW264. 7 macrophages，while KPA upregulated the expression of Prdx6．Moreover，treat- ment with the Prdx6 inhibitor MJ33 significantly upregulated the expression of iNOS，a marker of M1 macrophage polarization，in RAW264. 7 macrophages，whereas treatment with KPA significantly downregulated the expression of iNOS ( all P＜0. 05) ．Conclusion KPA inhibits LPS-induced M1 polarization of RAW264. 7 macrophages by up- regulating the expression of Prdx6．

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To investigate the effect of loss of function of lanosterol synthase( LSS) gene on intestinal function in a mouse model of non-alcoholic fatty liver disease( NAFLD) induced by a high-fat diet．Methods LSS gene heterozygous knockout C57 mice ( LSS + / －) were established using the CRISRP / Cas9 system．After being fed a high-fat diet with 60% fat content for 6 months，the fat deposition in liver tissues was detected by HE and Oil red O staining，the morphological changes of small intestine tissue were detected by HE staining．The changes in total cholesterol content in intestinal tissue were detected by kits．The gastrointestinal motility function of mice was de- tected by phenol red paste．The intestinal permeability was detected by Evans blue staining，and the expression of LSS，tight junction protein ( Claudin) -1，Claudin-5，cluster of differentiation 36 ( CD36) ，and Niemann-Pick type C1-like 1 protein ( NPC1L1) proteins in small intestinal tissues were detected by Western blot．Results The re- sults of HE and Oil red O staining of liver tissues showed that liver fat deposition in LSS gene heterozygous knockout mice was lower than that in wild-type mice in the high-fat diet group．The total cholesterol content in intestinal tis- sue of LSS gene heterozygous knockout mice decreased ( P ＜0. 01) ，but no morphological differences were ob- served between the two groups of mice by HE staining of intestinal tissues．The gastrointestinal motility function of LSS gene heterozygous knockout mice did not show significant changes．The intestinal permeability of LSS gene het- erozygous knockout mice in the high-fat diet group decreased as detected by Evans blue ( P＜0. 05) ．The expres- sion levels of Claudin-5 protein in the intestinal tissue of LSS gene heterozygous knockout mice in the high-fat diet group increased ( P ＜0. 05 ) ，while the expression of LSS protein in the intestinal tissues of LSS heterozygous knockout mice decreased ( P ＜0. 05) ．Conclusion In the NAFLD model induced by a high-fat diet，LSS gene heterozygous knockout reduces liver fat deposition induced by a high-fat diet and improves intestinal barrier function by regulating cholesterol metabolism in intestinal tissues and up-regulating the expression of Claudin-5．

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To investigate the impact of mandibular retrognathism on brain function in rats and its poten- tial mechanisms．Methods Male SD rats aged 3 to 4 weeks ( w) were randomly divided into control group ( n = 10) and mandible retrusion ( MR) group ( n = 30) ，with the MR group further divided into three subgroups: 1，2， or 4 weeks，with 10 rats in each subgroup．The MR group used modified metal tubes，which were bonded to the lingual side of the maxillary incisors in rats，forcing the mandible to retract by 2－2. 5 mm，thereby achieving the goal of constructing a mandibular retrognathism model; the control group received no interventions．The effects of mandibular retraction on the spatial learning ability and memory ability of rats were explored by behavioral experi- ments such as Morris water maze，Y maze and new object recognition 1，2，and 4 weeks after modeling，respective- ly．Hematoxylin-eosin ( HE) and Nissl staining were employed to observe the histomorphological changes in CA1， CA3，and DG regions of hippocampal tissues．Immunohistochemistry was employed to detect the protein expression levels of tropomyosin receptor kinase B ( TrkB) in the CA1，CA3，and dentate gyrus ( DG) regions of the hippo- campus．Western blot was used to assess the expression changes in the TrkB / phosphoinositide 3-kinase ( PI3K) / protein kinase B ( AKT) pathway in hippocampal tissue．Quantitative analysis of apoptosis-related proteins in the hippocampus，including Bcl-2-associated X protein ( Bax) ，B-cell lymphoma-2 protein ( Bcl-2) ，and Caspase-3， was conducted using qRT-PCR and Western blot．Results Behavioral experiments showed that rats in the MR group，especially the MR 4 w group，were significantly weaker in learning and spatial memory than the control group，MR 1 w，and MR 2 w( P＜0. 05) ．HE staining showed that pyramidal cells in the control group were ar- ranged orderly with regular morphology．In contrast，the MR group exhibited a reduction in the number of cells in the hippocampal CA1，CA3，and dentate gyrus ( DG) regions．The cells were sparsely and irregularly arranged， with nuclear condensation and enlarged cellular spaces． Nissl staining further demonstrated damage to the Nissl bodies in the MR group．Meanwhile，the TrkB / PI3K / AKT signaling pathway was activated in the MR group，and the expression of p-TrkB，PI3K，and p-AKT proteins was up-regulated ( P ＜0. 05) ．Western blot and qRT-PCR indicated that MR up-regulated the expression of pro-apoptotic protein Bax，apoptotic effector protein Caspase-3 ( P＜0. 05) ，and inhibited the expression of anti-apoptotic protein Bcl-2 ( P ＜0. 05) ．Conclusion Mandibular retraction impairs brain function and affects cognition and learning memory in rats．This may be achieved by regula- ting the TrkB / PI3K / AKT signaling pathway to induce apoptosis in hippocampal neurons．

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To investigate the role of STAT3 phosphorylation in ferroptosis regulation and its impact on cisplatin resistance mechanisms in human osteosarcoma cells．Methods Human osteosarcoma HOS cells and cispl- atin-resistant HOS / DDP cells were treated with cisplatin ( 0. 5，1，2，4，8，16，32 mg / L) ，the ferroptosis inducer Erastin ( Era) ( 2 μmol / L) ，and / or the ferroptosis inhibitor Ferrostatin-1 ( Fer1) ( 10 μmol / L) ．Cell viability and proliferation were assessed using the Cell Counting Kit-8 ( CCK-8) and colony formation assays，and cell migration was evaluated via a scratch assay．Reactive oxygen species ( ROS) ，intracellular ferrous iron levels，mitochondrial membrane potential，mitochondrial function，malondialdehyde ( MDA) levels，and the reduced glutathione / oxi- dized glutathione ( GSH / GSSG) ratio were measured using commercial kits．The mRNA expression of ferroptosis- related genes was analyzed by quantitative reverse transcription polymerase chain reaction ( RT-qPCR) ．The protein levels of glutathione peroxidase 4 ( GPX4) ，solute carrier family 7 member 11 ( SLC7A11) ，phosphorylated STAT3 ( p-STAT3) ，and total STAT3 were determined by Western blot．Results With cisplatin treatment，HOS cells ex- hibited decreased cell viability，mitochondrial membrane potential，and GSH / GSSG ratio ( P＜0. 01) ，along with elevated levels of ROS，ferrous ion，and MDA content ( P ＜0. 01) ． The protein levels of GPX4 ( P ＜0. 01) ， SLC7A11，and p-STAT3 also decreased ( P ＜0. 05) ．Coadministration with the ferroptosis inhibitor Ferrostatin-1 ( Fer-1) reversed these aforementioned effects ( P ＜0. 05) ．In HOS / DDP cells，the mRNA levels of ferroptosis- suppressive genes ( GPX4，FTH1，SLC7A11，and AIFM2) were significantly higher than those in HOS cells ( P < 0. 05) ，whereas the expression of ferroptosis-promoting genes ( ACSL4 and PTGS2) was significantly lower ( P < 0. 05) ．The cisplatin-induced reductions in cell viability and mitochondrial membrane potential，as well as the in- creases in ROS，ferrous ion，and MDA levels，were less pronounced in HOS / DDP cells than in HOS cells．The SLC7A11 protein level showed no significant change． However，combined treatment with the ferroptosis inducer Erastin ( Era) resulted in significant decreases in viability，mitochondrial membrane potential，and the GSH / GSSG ratio in HOS / DDP cells ( P＜0. 05) ．Furthermore，the protein levels of p-STAT3，GPX4，and SLC7A11 were also markedly reduced ( P＜0. 05) ．Conclusion The activation of ferroptosis mediated by p-STAT3 enhances cisplatin sensitivity in HOS / DDP cells．

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To explore the effect of interferon regulatory factor 2 binding protein 2 ( IRF2BP2) on the proliferation of acute myeloid leukemia ( AML) cells and its molecular mechanism．Methods The CRISPR-Cas9 gene editing technology was used to knock out IRF2BP2 in human AML cell lines Kasumi-1 and U937，and West- ern blot was performed to detect the knockout efficiency of IRF2BP2 protein．Cell morphology was observed using a microscope．Cell phenotypes were analyzed by CCK-8 assay，colony formation experiments，and flow cytometry． RNA-Seq was performed to identify differentially expressed genes between the IRF2BP2 knockout group and the control group in the U937 cell line．Gene Set Enrichment Analysis ( GSEA) was conducted to explore the down- stream molecular mechanisms．Western blot was used to detect the expression of downstream differentially expressed genes．The Cleavage Under Targets and Tagmentation ( CUT＆Tag) technique was applied to identify the direct tar- gets of the IRF2BP2 protein，and the corresponding binding signals were visualized using the Integrated Genomics Viewer ( IGV) ．Results Compared with the control group，after knocking out IRF2BP2，the CCK-8 experiment showed that AML cell proliferation was inhibited ( P ＜0. 05) ; the number of colonies in the IRF2BP2 knockout group decreased ( P＜0. 05) ，and the proportion of G1 phase was prolonged ( P＜0. 05) ; in U937 cell lines，knoc- king out IRF2BP2 resulted in significant enrichment of differential genes in myelocytomatosis oncogene ( MYC) -re- lated signaling pathways，and the protein expression levels of pathway molecules MYC，cyclin-dependent kinase 4 ( CDK4) ，and cyclin － dependent kinase 2 ( CDK2 ) decreased with the downregulation of IRF2BP2; using IRF2BP2 antibodies in U937 cell lines for CUT＆Tag experiments，IGV visualization analysis showed a significant increase in signal peaks in the MYC promoter region．Conclusion IRF2BP2 protein affects the cell cycle and pro- liferation of AML cells by targeting and regulating MYC．

abstract:
To identify autophagy-related genes involved in pulmonary infection caused by the highly drug-resistant and virulent methicillin-resistant Staphylococcus aureus strain USA300 ( USA300) ，and to explore the underlying molecular mechanisms ， thereby providing potential targets for immunotherapy． Methods The GSE220943 dataset of a USA300-induced pulmonary infection mouse model was obtained from the GEO database． Differentially expressed genes ( DEGs ) were identified using the DESeq2 package． Autophagy-related genes ( ARGs) were retrieved from the MSigDB and Autophagy databases．Weighted gene co-expression network analysis ( WGCNA) was performed to construct gene co-expression modules．Genes overlapping among DEGs，ARGs，and WGCNA modules were identified as autophagy-related DEGs．Gene Ontology ( GO) enrichment analysis was con- ducted using the clusterProfiler R package，while Kyoto Encyclopedia of Genes and Genomes ( KEGG) pathway en- richment analysis was performed via the Metascape platform．Immune cell infiltration was analyzed using the Immu- CellAI-mouse website．A protein - protein interaction ( PPI) network was constructed using the STRING database， and hub genes were identified through topological analysis in Cytoscape． Receiver operating characteristic curve ( ROC) curves were plotted via the website https: / /www．bioinformatics．com．cn． Finally，key gene expression was validated in mouse lung tissues by real-time quantitative reverse transcription PCR ( RT-qPCR) ．Results A total of 6 135，4 075，3 680，and 2 342 differentially expressed genes ( DEGs) were identified at 12，24，48，and 96 hours post-infection，respectively．By integrating DEGs，autophagy-related genes ( ARGs) ，and WGCNA mod- ules，19 autophagy-related DEGs were identified． GO and KEGG enrichment analyses indicated that these genes were mainly involved in CD4 + T cell activation and regulation，innate immune responses，and autophagosome mem- brane formation．Immune infiltration analysis revealed that innate immune cells such as neutrophils and dendritic cells predominated during the early phase of infection，while γδ T cells and M2 macrophages became more promi- nent in the later stages．PPI network analysis identified 12 hub autophagy-related genes，among which three upreg- ulated key genes ( Eif2ak2，Ikbke，and Nfkbiz) were further confirmed．The area under the ROC curve for all three genes was 1. 000．RT-qPCR validation demonstrated significantly elevated expression of these three genes in lung tissues at 24 hours post-infection ( all P＜0. 05) ．Conclusion Eif2ak2，Ikbke，and Nfkbiz may be involved in the pulmonary infection caused by USA300 by promoting autophagy and hold promise as potential targets for immuno- therapy．

abstract:
Objective To explore the therapeutic effect of extracellular vesicles(EVs) of Aspergillus flavus(A. flavus) on A. flavus infection. Methods The EVs of A. flavus were isolated using ultracentrifugation and detected/identified by nanoparticle tracking analysis(NTA). Quantitative real-time PCR(qRT-PCR) and enzyme-linked immunosorbent assay(ELISA) kits were used to assess the effect of A. flavus EVs on the polarization of bone marrowderived macrophages(BMDMs) and the expression levels of several cytokines,including tumor necrosis factor-α(TNF-α),interferon-γ(IFN-γ),interleukin-6(IL-6),and interleukin-10(IL-10). The Galleria mellonella(G.mellonella) infection model was constructed. Three concentration groups of A. flavus EVs(0. 2,2,and 20 μg/larvae) were set as the experimental groups,and PBS was used as the control group for the infection model.Meanwhile,three forms of negative control groups were established,including the PBS group,the pierced controlgroup, and the negative control group. The therapeutic effect of A. flavus EVs on A. flavus infection was evaluated by the survival rate of the G. mellonella infection models. Results The particle size of A. flavus EVs ranged from 20 to550 nm. A. flavus EVs could polarize BMDMs into both M1 and M2 phenotypes and induce the production of cytokines,including TNF-α,IFN-γ,IL-6,and IL-10. The results of the G. mellonella infection model showed that A.flavus EVs could improve the survival rate of G. mellonella after A. flavus infection. Conclusion The EVs produced by A. flavus can promote the expression of both pro-inflammatory and anti-inflammatory cytokines in BMDMs,induce M1 polarization and M2 polarization of BMDMs,and increase the survival rate of G. mellonella after A. flavus infection.

abstract:
To investigate the effect of receptor of activated protein kinase C 1 ( RACK1) on lipopo- lysaccharide ( LPS) -induced barrier function in rat pulmonary microvascular endothelial cells ( RPMVEC) and its interplay with the Sonic hedgehog ( SHH) signaling pathway．Methods RPMVEC were cultured in vitro and ran- domly divided into si-NC，si-NC + LPS，si-RACK1，si-RACK1 + LPS，si-RACK1 + LPS + Vismodegib and Vis- modegib + SAG groups．The RACK1 of RPMVEC was silenced by small interfering RNA ( siRNA) technology and the cells were treated with LPS ( 10 mg / L) ，SHH signaling pathway inhibitor ( Vismodegib) ( 20 μmol / L) and SHH signaling pathway agonist ( SAG) ( 1 μmol / L) ．Following the intervention，the expressions of RACK1 and caveolin-1 in RPMVEC were detected by immunofluorescence ，while the transendothelial electrical resistance ( TEER) was evaluated using the method of Transwell，and the expression levels of RACK1，glioma-associated on- cogene homolog 1 ( Gli-1) and caveolin-1 were detected by Western blot．Results Silencing RACK1 increased the TEER value of RPMVEC induced by LPS ( P＜0. 05) ．The expression of caveolin-1 decreased ( P＜0. 05) ，and the expression of Gli-1 increased ( P ＜0. 05) ．Inhibiting the SHH signaling pathway could reverse the increased TEER value of LPS-induced RPMVEC caused by silencing RACK1 ( P ＜0. 05) ，and the expressions of RACK1 and caveolin-1 increased ( P＜0. 05) ．Activation of the SHH signaling pathway increased the TEER value of LPS- induced RPMVEC caused by silencing RACK1 ( P ＜0. 05) ，and the expressions of RACK1 and caveolin-1 de- creased ( P＜0. 05) ．Conclusion RACK1 plays a role in LPS-induced hyperpermeability of RPMVEC，and its effect may be achieved by modulating the SHH signaling pathway and caveolin-1．

abstract:
Objective To investigate the regulatory effect of corticosterone(CORT) pretreatment on the activation of microglia by lipopolysaccharide(LPS) and the inhibitory effect of Biochanin A(Bio A) on microglia activation.Methods The MTT method was used to select the optimal concentrations for LPS,CORT and Bio A on BV2 cells;BV2 cells were divided into 5 groups: Control group,CORT(50 nmol/L) group,LPS(1 μg/mL) group,LPS(1μg/mL) + CORT(50 nmol/L) group,and LPS(1 μg/mL) + CORT(50 nmol/L) + Bio A(5 μmol/L) group;Except for the control group,each group was first incubated with CORT(50 nmol/L) for 2 h,and then each group was co-incubated with the corresponding concentrations of LPS(1 μg/mL) and Bio A(5 μmol/L) for 36 h;DCFH-DA probe method was used to detect reactive oxygen species(ROS) content; Western blot was used to detect the protein expression levels of inflammatory cytokines,5-hydroxytryptamine(5-HTT),glucocorticoid receptor(GR),mineralocorticoid receptor(MR),NOD-like receptor family pyrin domain containing 3(NLRP3),apoptosis-associated speck-like protein containing a CARD(ASC) and cysteine-aspartic protease 1(Caspase-1). Results Compared with the CORT(50 nmol/L) and LPS(1 μg/mL) groups,cells in the CORT(50 nmol/L) + LPS(1 μg/mL) group showed increased cell viability,higher levels of ROS,and increased levels of inflammatory cytokines,NLRP3,ASC,and Caspase-1 protein expression( P＜0. 05 ) ．Conclusion A low dose of CORT pretreatment reinforces LPS- induced BV2 cell activation; Bio A inhibits CORT-pretreated LPS-induced BV2 cell activation．The mechanism of which may be related to the inhibition of NLRP3 inflammasome activation．

abstract:
To investigate the high-risk human papillomavirus ( HR-HPV) infection and its relationship with vaginal infectious diseases among women and to provide a basis for the prevention and control of cervical canc- er among women in this region．Methods Uterine exfoliative cells and vaginal secretions were collected from women aged 25－64 years in the resident population ( ≥6 months of residence) who participated in cervical cancer screening and human papillomavirus typing and vaginal microecology tests were performed to compare the status of HPV infections in different age groups．The study subjects were divided into HR-HPV-positive and HR-HPV-nega- tive groups according to whether HR-HPV infection was detected or not，and multifactorial Logistic regression anal- ysis was used to explore the relationship between HR-HPV and vaginal infectious diseases．Results The HR-HPV infection rate of women in this study was 10. 43% ，and the HR-HPV infection rate of each age group was 14. 34% from 25 to 34 years old，10. 16% from 35 to 44 years old，8. 70% from 45 to 54 years old，and 11. 89% from 55 to 64 years old，and there were differences in the infection rate of HR-HPV among different age groups ( P < 0. 05) ．Among HR-HPV，HPV52，HPV16，HPV58，HPV51，HPV68 were the most common，with 76. 69% of sin- gle infections，18. 02% of dual infections，4. 46% of triple infections，and 0. 83% of quadruple infections．HR- HPV single infections and multiple infections were prevalent in the age group of 25－34 years old and 55－64 years old．HR-HPV mono-infection and multi-infection rates were statistically different between age groups ( P＜0. 05) ． The infection rates of bacterial vaginosis ( BV) and trichomoniasis vaginitis ( TV) were higher in HR-HPV-positive patients ( P ＜0. 05) ． Logistic regression analysis showed that the infection rates of BV ( OR = 1. 560，95% CI: 1. 195－2. 035 ) ，TV ( OR = 2. 208，95% CI: 1. 221 －3. 993 ) were risk factors for HR-HPV infection ( P < 0. 05) ．Conclusion In the screened population of the Fourth Division of Xinjiang Production and Construction Corps，HR-HPV mono-infection is the most prevalent type．Women aged 25－34 and 55－64 years old show higher rates of HR-HPV infection．The most common HR-HPV genotypes，in descending order，are 52，16，58，51，and 68．BV and TV are the two major risk factors closely associated with HR-HPV infection．

abstract:
Objective To evaluate the clinical efficacy of preimplantation genetic testing for monogenic disorders(PGT-M) in families with hereditary epilepsy. Methods Whole-exome sequencing(WES) and familial co-segregation analysis were performed to validate the pathogenicity of variants(PCDH19 c. 1031C > G and LGI1 c. 856T >G) in two monogenic epilepsy families. A clinical PGT-M pathway was implemented,and reproductive outcomes were tracked. Results In Family 1(PCDH19 likely pathogenic variant),13 blastocysts were biopsied over two ovarian stimulation cycles,yielding 3 unaffected euploid embryos(23. 1%). After the third frozen embryo transfer,a healthy male infant was successfully delivered. Prenatal diagnosis confirmed that the fetus did not carry the pathogenic variant PCDH19. Family 2(LGI1 variant of uncertain significance,VUS) screened 14 blastocysts,identifying 2 unaffected euploid embryos(14. 3%),with the first transfer unsuccessful. A clinical pregnancy was currently ongoing following the second frozen-thawed embryo transfer(FET). Conclusion PGT-M can precisely block the vertical transmission of monogenic epileptic pathogenic variants,offering an effective reproductive intervention strategy for families with hereditary epilepsy.

abstract:
To assess the risk factors for symptomatic intracranial hemorrhage ( sICH) after endovascu- lar therapy ( EVT) in patients with acute anterior circulation large vessel occlusion．Methods A retrospective a- nalysis was used to analyze the clinical data of 273 anterior circulation large vessel occlusion ( ACLVO) patients with EVT，of which 158 were male and 115 were female; ages ranged from 33 to 94 years old［71 ( 59，78) years old］，and sICH occurred in 61 cases．The patients' gender，age，history of smoking and alcohol consumption，his- tory of illness，history of oral anticoagulant / anti-antibiotic administration，oral anticoagulant / antiplatelet medication status，intravenous thrombolytic therapy，admission blood pressure，hospitalization days，occlusion location，proce- dure duration，number of times of thrombus retrieval，cause of stroke ( TOAST classification) ，revascularization status ( mTICI ≥2b) ，and admission ASPECT score，admission APACHEII score，and admission NHISS score were collected．Univariate analysis was used to compare the differences in the above indicators between the non- SICH group and the sICH group． Multivariable Logistic regression analysis was used to evaluate the independent factors for the occurrence of sICH after EVT．Results The results of univariate analysis showed that mTICI ≥ 2b， admission ASPECT score，and post-EVT sICH were significantly negatively correlated ( P＜0. 05) ，while occlusion location ( ICA) ，admission APACHE-Ⅱ score，admission NHISS score＞20，and admission mRS score were sig- nificantly positively correlated with post-EVT sICH ( P＜0. 05) ．The results of the multivariable Logistic regression analysis indicated that mTICI≥2b ( OR = 0. 314，95% CI: 0. 106－0. 929) and admission ASPECT score ( OR = 0. 775，95% CI: 0. 604－0. 995) are protective factors for sICH after EVT，while occlusion location ( ICA) ( OR = 2. 047，95% CI: 1. 033－4. 054) and admission APACHE-Ⅱ score ( OR = 1. 236，95% CI: 1. 140 －1. 340) were risk factors for sICH after EVT．Conclusion Higher admission APACHEII score，lower admission ASPECT score，occlusion location ( ICA) ，and mTICI ＜2b are independent risk factors for the development of sICH after EVT in patients with acute ACLVO．

abstract:
Objective To construct and to validate a nomogram prediction model based on Lipoprotein-associated phospholipase A2(Lp-PLA2) and Lipoprotein(a) [LP(a) ]for predicting the risk of major adverse cardiovascular events(MACE) in patients with coronary heart disease(CHD). Methods A retrospective analysis was conducted on the clinical data of 442 patients with coronary heart disease(CHD). Among them,411 patients who completed follow-up were randomly divided into a training set(288 cases) and a validation set(123 cases) at a 7 ∶ 3 ratio.Independent risk factors for major adverse cardiovascular events(MACE) in CHD patients were screened through Lasso regression analysis and Cox regression analysis,and a nomogram prediction model was constructed. The predictive performance of the model was evaluated using time-dependent receiver operating characteristic curves(ROC),calibration curves,and decision curve analysis. Results Variables were screened through Lasso regression and Cox regression analysis. The final model included nine independent predictors,namely age,smoking history,clinical phenotype of CHD,the number of coronary artery lesions,Gensini score,BNP,Lp-PLA2,LP(a), and the history of statin use. The area under the ROC curve in the training set was 0. 897,0. 885,and 0. 909 at 1,2,and 3 years,respectively; The area under the ROC curve in the validation set was 0. 885,0. 881,and 0. 923 at 1,2,and 3 years,respectively. These results demonstrated that the model had excellent discriminatory power. The calibration curves and decision curves demonstrated that the model had high clinical practicality in predicting the occurrence of MACE in CHD patients. Conclusion The nomogram prediction model based on LP-PLA2,LP(a)and other risk factors provides an effective tool for the prognosis assessment of CHD patients,facilitating the early identification of high-risk patients and enabling individualized intervention.

abstract:
To explore the value of nomogram based on DCE-MRI radiomics combined with clinical-ra- diological features in predicting HR status in breast cancer with Her-2 low expression．Methods A total of 198 pa- tients of Her-2 low expression breast cancer who underwent standardized breast MRI in our hospital from January 2019 to February 2025 were retrospectively analyzed．Patients were divided into HR ( + ) group ( n = 137) and HR ( －) group ( n = 61) ．The cases were divided into a training set ( 138 cases) and a testing set ( 60 cases) in a 7 ∶ 3 ratio．Clinical-radiological model was based on clinical and traditional radiological features，radiomics model was based on DCE-MRI，and combined model was constructed，respectively．The nomogram was drawn，and re- ceiver operating characteristic curve was used to compare the performance of different models in predicting HR sta- tus．Results The DCE-MRI radiomics score ( Radscore) between the HR ( + ) group and the HR ( －) group showed statistical differences in both the training and testing sets ( both P＜0. 001) ．The AUC of the clinical-radio- logical model based on lesion mobility，Ki67，TIC type，enhancement pattern and maximum diameter for predicting HR status in the training set and testing set were 0. 643 and 0. 616，respectively．The AUC of the DEC-MRI ra- diomics model in the training set and testing set were 0. 897 and 0. 860，respectively．The nomogram drawn by combining clinical-radiological features and Radscore showed better predictive performance in both the training set ( AUC = 0. 913) and testing set ( AUC = 0. 898) than the clinical-radiological model ( all P＜0. 05) ．Conclusion The nomogram combined by DCE-MRI radiomics and clinical-radiological features can effectively predict HR sta- tus of breast cancer with low Her-2 expression，which is helpful to the building of individualized treatment plan for breast cancer patients．

abstract:
To investigate the diagnostic value of combined detection of folate receptor-positive circulat- ing tumor cells ( FR + -CTCs) and hemostatic function indicators in improving the diagnosis of gastrointestinal tumors ( GITs) metastasis．Methods A retrospective analysis was conducted on the clinical data of 115 patients aged 18 to 80 years who were diagnosed with gastrointestinal tumors via pathology and received treatment，including data on FR + -CTCs，hemostatic function indicators，and pathological staging．The collected data encompassed FR + -CTCs levels，coagulation parameters，and pathological staging．Statistical analysis included t-tests，chi-square tests，fish- er ’s exact test，Logistic regression analysis，and receiver operating characteristic ( ROC) curves to assess the diag- nostic value of combined FR + -CTCs and coagulation parameters in detecting tumor metastasis．Results FR + -CTCs levels and positive rates demonstrated significant associations with clinicopathological characteristics ( gender，histo- logical type，N staging) in GITs patients ( P＜0. 05) ．In patients with metastasis，elevated fibrinogen levels were observed．Adithonallly，platelet counts showed significant increases in N1 －N3 stages ( P＜0. 05) ．Logistic regres- sion analysis showed that PLT and antithrombin Ⅲ ( AT-Ⅲ) were independent risk factors for GITs metastasis ( P＜0. 05) ． The areas under the ROC curves for predicting GITs metastasis were 0. 678 ( 95% CI: 0. 540 - 0. 816) and 0. 664 ( 95% CI: 0. 512 －0. 815) ，respectively．When combining multiple factors，including FR + - CTCs，PLT，AT-Ⅲ , pathological type，FIB，TT，and gender，for the diagnosis of GITs metastasis，the AUC in- creased to 0. 757 ( 95% CI: 0. 621 －0. 893) ，indicating higher sensitivity and specificity compared to using each indicator alone．Conclusion The combined detection of FR + -CTCs and anticoagulation function indicators has a higher diagnostic value for the diagnosis of GITs，providing a valuable basis for the early diagnosis of GITs，espe- cially in metastasis surveillance．

abstract:
To explore the value of combined indicators of prognostic nutritional index ( PNI) ，nutri- tional risk screening 2002 scale ( NRS2002 ) ，body mass index ( BMI) ，monocyte / lymphocyte ratio ( MLR) ， platelet / lymphocyte ratio ( PLR) ，and neutrophil / lymphocyte ratio ( NLR) in evaluating the severity of pulmonary tuberculosis ( PTB) ．Methods A total of 175 patients with pulmonary tuberculosis were selected as the study group，and 175 healthy subjects who underwent physical examination during the same period were selected as the control group．According to the range of lung lesions，the patients were divided into mild to moderate group ( ＜3 lung fields，n = 110) and severe group ( ≥3 lung fields，n = 65) ，PNI，NRS2002，BMI and peripheral blood MLR , PLR and NLR levels were compared between the two groups．Spearman rank correlation was used to analyze their correlation with the severity of the disease．A multivariate logistic regression model was established．A nomo- gram was drawn，and the efficiency of the model was evaluated by receiver operating characteristic ( ROC) curve， calibration curve，decision curve．Results The PNI，BMI，peripheral blood lymphocyte and albumin levels in the study group were lower than those in the control group ( P＜0. 05) ，while the MLR , PLR and NLR levels were sig- nificantly higher than those in the control group ( P＜0. 05) ．The NRS2002，MLR , PLR and NLR levels in the se- vere group were higher than those in the mild and moderate group ( P＜0. 05) ，while the PNI and BMI levels were lower than those in the mild and moderate group ( P＜0. 05) ．NRS2002 and peripheral blood MLR , PLR and NLR levels in patients with pulmonary tuberculosis were positively correlated with the severity of the disease ( r = 0. 250， 0. 509，0. 431 and 0. 488) ．PNI and BMI were negatively correlated with the severity of the disease ( r = －0. 516， - 0. 231) ．Multivariate Logistic regression showed that NRS2002 and NLR were independent risk factors for severe disease，while PNI was a protective factor．The areas under the curve ( AUC) of NRS2002，NLR , PNI，and com- bined detection for evaluating the severity of pulmonary tuberculosis were 0. 692，0. 777，0. 786，and 0. 860，re- spectively，the sensitivity of the combined prediction was 81. 54% and the specificity was 76. 36%．The combined detection had better evaluation efficiency for severe pulmonary tuberculosis than the single indicator ( P ＜0. 05) ． Conclusion NRS2002 and NLR are independent risk factors for severe disease，while PNI is a protective factor． The combined detection model has a good fit，which can improve the evaluation efficiency and has potential for clin- ical application．

abstract:
Ferroptosis is a novel form of cell death characterized by the iron-dependent accumulation of lipid hydroperoxides. Since it was first proposed in 2012,Ferroptosis has gradually attracted attention and developed rapidly. Ferroptosis plays an important role in cardiovascular diseases,malignant tumors and neurological diseases,and has become a research hotspot in the field of life science and medicine. Ferroptosis is closely related to iron overload. Iron overload and the accumulation of lipid peroxidation jointly contributes to the disruption of intervertebral disc homeostasis,leading to intervertebral disc degeneration. However,the specific mechanisms of ferroptosis in regulating intervertebral disc degeneration is not yet clear. This review discusses the relationship between ferroptosis and intervertebral disc degeneration,their molecular regulatory mechanisms,and their potential clinical applications,aiming to provide new therapeutic targets for intervertebral disc degeneration.