Abstract:
Objective　To investigate the inhibitory effect of gingerol， an active component of ginger， on Staphylococcus aureus （S. aureus） infection， and to preliminarily explore its mechanism related to extracellular vesicles （EVs）.Methods　S.aureus infection models were established in human umbilical vein endothelial cell （HUVECs）， Vero E6 cells， and C57BL/6 mice. Experimental groups included control， infection， and gingerol-treated groups. Bacterial load and VE-cadherin protein expression were detected using immunofluorescence and Western blot. EVs were isolated by size exclusion chromatography and characterized by transmission electron microscopy and nanoparticle tracking analysis. miRNA sequencing was performed on EVs.Results　Gingerol treatment significantly reduced the bacterial load in both in vitro and in vivo infection models and upregulated VE-cadherin expression. miRNA sequencing of EVs revealed that S. aureus infection upregulated the expression of hsa-miR-320c， while gingerol treatment reversed this abnormal expression. Bioinformatic analysis further predicted that the target genes of hsa-miR-320c were significantly enriched in cell junction-related pathways. Conclusion　Gingerol exhibits clear antibacterial and host-protective effects， by regulating hsa-miR-320c in EVs to maintain endothelial barrier integrity.

Abstract:
Objective　To investigate the effect of protein arginine methyltransferase 5 （PRMT5） on the angiogenesis of colorectal cancer （CRC） through the Janus kinase 2/signal transducer and activator of transcription 3 （JAK2/STAT3） signaling pathway.Methods　The expression and prognostic value of PRMT5 in CRC tissues were analyzed based on the cancer genome atlas （TCGA） and gene expression omnibus （GEO） databases. A total of 105 pairs of cancer tissues and matched adjacent non-tumor tissues were collected from CRC patients， and PRMT5 expression was detected by immunohistochemistry （IHC）； Western blot was used to detect the PRMT5 protein level in normal colonic epithelial cells （FHC） and CRC cell lines （including RKO， SW480）. SW48 and LoVo cells were divided into negative control （NC） group and PRMT5 knockdown interference group. After the stable knockdown cell models were successfully constructed， the effects of conditioned medium on the proliferation， cell cycle， migration and tube formation abilities of human umbilical vein endothelial cells （HUVECs） were detected； the correlation between PRMT5 and vascular endothelial growth factor A （VEGFA） was analyzed by real-time quantitative polymerase chain reaction （RT-qPCR） and enzyme-linked immunosorbent assay （ELISA）. Gene set enrichment analysis （GSEA） and cytoplasm-nucleus separation assay were performed to verify the regulatory pathway.Results　PRMT5 was highly expressed in CRC tissues and cell lines， which was significantly correlated with poor prognosis and pathological characteristics of patients （all P<0.05）； PRMT5 knockdown could significantly inhibit the angiogenesis-related abilities of HUVECs （all P<0.05） and down-regulate the expression of VEGFA （all P<0.01）. GSEA results indicated that PRMT5 expression was significantly associated with the activation of JAK2/STAT3 signaling pathway （all P<0.001）. PRMT5 knockdown could inhibit the activation of JAK2/STAT3， the nuclear translocation of STAT3 and the protein level of phosphorylated STAT3 （p-STAT3） in the nucleus， thus inhibiting the expression of VEGFA.Conclusion　PRMT5 is highly expressed in CRC tissues and indicates poor prognosis of patients. It promotes the nuclear translocation and transcriptional activity of STAT3 by activating the JAK2/STAT3 pathway and drives the expression of VEGFA to promote tumor angiogenesis.

Abstract:
Objective　To investigate whether intestinal deficiency of lanosterol synthase （LSS）， a key enzyme in cholesterol synthesis， influences the progression of metabolic dysfunction-associated steatotic liver disease （MASLD） by regulating intestinal cholesterol absorption and immune response.Methods　LSS heterozygous knockout （LSS+/-） mice and wild-type （WT） controls were generated using CRISPR/Cas9 technology and fed either a high-fat diet （HFD） or regular chow （CHOW）. The model was validated by genotyping. Hepatic steatosis was assessed by HE and oil red O staining. Immunohistochemistry was used to detect the localization and expression of NPC1L1 and CD36 proteins in the intestine. Western blot analysis was performed to measure JNK phosphorylation and TLR4 protein levels in intestinal tissues. Real-time quantitative polymerase chain reaction （qPCR） was employed to examine the mRNA expression of TLR4 and IL-6.Results　LSS+/- mice were successfully validated by genotyping and reduced intestinal LSS protein expression. HE and oil red O staining of liver sections showed that， compared with WT mice fed a CHOW diet， WT mice fed a HFD exhibited a marked increase in hepatic lipid vacuoles. In contrast， compared with HFD-fed WT mice， HFD-fed LSS+/- mice displayed significantly attenuated hepatic lipid deposition and reduced serum ALT levels （P<0.05）.Immunohistochemical analysis revealed that， compared with WT mice， the expression of the cholesterol absorption protein NPC1L1 in the intestinal villi of LSS+/- mice was downregulated under both CHOW and HFD conditions （PHFD<0.001）. Conversely， the expression of the fatty acid transporter CD36 was upregulated in the intestines of LSS+/- mice （PCHOW<0.05， PHFD<0.01）. Western blot analysis demonstrated that， compared with WT mice， TLR4 protein expression in the intestines of LSS+/- mice significantly increased under both CHOW and HFD conditions （both P<0.05）. JNK phosphorylation level was significantly elevated in LSS+/- mice under CHOW condition （both P<0.05）. Under HFD condition， total JNK protein expression increased， but its phosphorylation level showed no significant change. qPCR analysis showed that， compared with WT mice， the mRNA levels of TLR4 （PCHOW<0.01， PHFD<0.000 1） and IL-6 （PCHOW<0.001， PHFD<0.01） were significantly upregulated in the intestines of LSS+/-mice.Conclusion　LSS deficiency counteracts hepatic lipid deposition by orchestrating a synergistic reprogramming involving restricted intestinal cholesterol absorption， enhanced fatty acid utilization， and activation of immune pathways， suggesting intestinal LSS as a potential therapeutic target of MASLD.

Abstract:
Objective　To investigate the effects of 7-dehydrocholesterol reductase （DHCR7） in promoting proliferation， migration， and invasion of lung adenocarcinoma cells and the underlying signaling mechanisms.Methods　The expression level of DHCR7 in lung adenocarcinoma and its prognosis were analyzed by using public databases. DHCR7 protein expression levels in the lung adenocarcinoma cell lines H1299 and A549 were assessed by Western blot， using the normal lung epithelial cell line BEAS-2B as a control. Small interfering RNA （siRNA） was used to knock down DHCR7 expression in H1299 and A549 cells. Assays including colony formation， CCK-8， wound healing， and Transwell experiments were conducted to assess the proliferation， migration， and invasion of the knockdown cells. Next， Western blot was employed to assess the phosphorylation levels of key signaling molecules in the phosphoinositide 3-kinase/protein kinase B （PI3K/AKT）， janus kinase/signal transducer and activator of transcription 3 （JAK/STAT3），mitogen-activated protein kinase/extracellular signal-regulated kinase pathway （MAPK/ERK） （p-AKT， p-STAT3， p-ERK）.Results　Bioinformatics results indicated that both DHCR7 mRNA and protein were highly expressed in lung adenocarcinoma compared with normal tissues adjacent to cancer. Furthermore， higher DHCR7 mRNA level was associated with poor prognosis （P<0.001）. The expression levels of DHCR7 were significantly higher in both H1299 and A549 cells than that in BEAS-2B cells （P<0.05， P<0.01）. Compared with si-control， the proliferation， migration， and invasion abilities of si-DHCR7 H1299 and A549 cells significantly decreased.Among the phosphorylated signaling molecules detected， p-ERK was significantly downregulated （P<0.000 1， P<0.01） whereas p-AKT and p-STAT3 levels were not significantly changed.Conclusion　　DHCR7 has prominent effects in promoting the proliferation， migration， and invasion of lung adenocarcinoma cells， and the underlying mechanisms are related to the MAPK/ERK signaling pathway.

Abstract:
Objective　To investigate the effect of honokiol on proliferation， apoptosis， migration， and invasion of gastric cancer cells and its underlying mechanistic.Methods　Human gastric cancer cell lines HGC-27 and AGS were treated with Honokiol at concentrations of 0， 15， and 25 μmol/L. CCK-8 assays were conducted to determine the half maximal inhibitory concentration （IC50） for both cell lines. Cell viability， proliferation， migration， and invasion capabilities were assessed using CCK-8， colony formation， wound healing， Transwell migration and Transwell invasion assays. Apoptosis rates were measured via flow cytometry. Western blot analysis examined proteins related to proliferation， apoptosis， migration， invasion， and the endoplasmic reticulum stress pathway ATF4-CHOP-TRIB3.Results　Compared with the control group， treatment with 15 and 25 μmol/L Honokiol significantly reduced the proliferation， colony formation， migration， and invasion capabilities of the two gastric cancer cell lines， while significantly increasing the apoptosis rate （P<0.05）. Additionally， compared to the control group， the protein expression levels of neural cadherin（N-cadherin）， Vimentin， proliferating cell nuclear antigen（PCNA）， and B-cell lymphoma/leukemia-2 protein（Bcl-2）decreased in the two gastric cancer cell lines after treatment with 15 and 25 μmol/L Honokiol， while the protein expression levels of epithelial cadherin（E-cadherin）， Bcl-2-associated X protein（Bax）， activating transcription factor 4（ATF4）， endoplasmic reticulum stress-related protein（CHOP）， and tribbles homolog 3（TRIB3）increased（P<0.05）.Conclusion　Honokiol promotes apoptosis and inhibits proliferation， migration， and invasion of gastric cancer HGC-27 and AGS cells by regulating the ERS signaling pathway ATF4/CHOP/TRIB3.

Abstract:
Objective　To investigate the expression changes of the Endophilin A1 （EPA1）-reactive oxygen species （ROS）-NLR family pyrin domain containing 1 （NLRP1） signaling pathway in chronic unpredictable mild stress （CUMS）-induced depression model mice.Methods　50 C57BL/6 mice were randomly divided into control group and CUMS model group（n=25）. The model mice received 42 days of CUMS stress exposure， after which behavioral changes were assessed through monitoring body weight， sucrose preference test， forced swim test， tail suspension test， open field test， and elevated plus-maze test. Hematoxylin-Eosin and Nissl staining were used to observe neuronal damage in hippocampal CA1 and CA3 regions. Calcium ion （Ca²⁺） assay kit was used to detect Ca²⁺ levels in the hippocampus. Immunofluorescence was used to detect colocalization of EPA1 and NLRP1 as well as ROS changes in the hippocampus. Transmission electron microscopy was used to observe mitochondrial structure in hippocampal neurons. The expression levels of calcium-activated neutral protease 1 （Calpain-1）， nicotinamide adenine dinucleotide phosphate oxidase 2 （NOX2）， NLRP1 inflammasome， downstream inflammatory proteins， and synapse-associated proteins in the hippocampus of mice was detected by using Western blot.Results　Compared with the control group， mice in the CUMS model group exhibited depressive-like behavior and hippocampal neuronal damage. The levels of NLRP1 and EPA1 significantly increased in the hippocampus， and both were co-expressed in the cytoplasm and membrane of hippocampal neurons in the CUMS group mice（P<0.01）. Ca²⁺ concentration was elevated（P<0.01）， and the protein levels of Calpain 1 and NOX2 were upregulated（P<0.01）. The average fluorescence intensity of ROS significantly increased， accompanied by structural damage to neuronal mitochondria（P<0.01）. The levels of NLRP1 and its downstream inflammatory proteins significantly increased （P<0.01）， while the expression levels of synapse-associated proteins significantly decreased （P<0.01）.Conclusion　EPA1 exhibits abnormal expression changes in CUMS-induced depression model mice， which may be closely associated with ROS generation， NLRP1 inflammasome activation， and the regulation of synaptic protein expression.

Abstract:
Objective　To evaluate the anti-inflammatory activity of the novel magnolol ester derivative YW and to investigate its effects on chondrocyte senescence and preliminary mechanisms.Methods　Magnolol and p-methylbenzoic acid were used as raw materials to synthesize the magnolol ester derivative YW （Molecular Formula： C26H24O3， Molecular Weight： 384.17， HPLC Purity >96%） via DCC/DMAP-catalyzed esterification. Cytotoxicity was assessed using the CCK-8 assay. A lipopolysaccharide （LPS）-induced RAW264.7 macrophage activation model and an interleukin-1β （IL-1β）-induced rat primary chondrocyte model were established. The release and mRNA expression of inflammatory factors including nitric oxide （NO）， IL-1β， tumor necrosis factor-alpha （TNF-α）， and IL-6 were detected by enzyme-linked immunosorbent assay （ELISA）， Griess reagent method， and quantitative real-time PCR （RT-qPCR）. The expression of senescence markers such as inducible nitric oxide synthase （iNOS）， pro-interleukin-1β （pro-IL-1β）， lysine acetyltransferase 7 （KAT7）， cyclin-dependent kinase inhibitor 1A （p21）， and cyclin-dependent kinase inhibitor 2A （p16）， as well as proteins related to chondrocyte extracellular matrix synthesis and catabolism， were analyzed by Western blot （WB）. Molecular docking was performed using Discovery Studio 2019 to validate target binding.Results　YW exhibited no significant cytotoxicity at concentrations ≤20 μmol/L. YW concentration-dependently inhibited LPS-induced macrophage inflammatory cytokine release， significantly downregulated iNOS， Pro-IL-1β protein， and inflammatory cytokine mRNA expression （P<0.01）. YW stably bound to KAT7 protein （binding energy： -94.2 kcal/mol）； YW downregulated KAT7 and aging marker protein expression in naturally aged and IL-1β-induced chondrocyte models （P<0.01）； YW regulated chondrocyte matrix synthesis and catabolic protein expression in IL-1β-induced chondrocytes （P<0.01）.Conclusion　YW inhibits macrophage activation and inflammatory cytokine release while downregulating KAT7 and senescence marker protein expression in chondrocytes， thereby blocking chondrocyte senescence.

Abstract:
Objective　To explore the expression levels of long non-coding RNA （lncRNA） H19 and microRNA-22-3p （miR-22-3p） in the serum of patients with acute myocardial infarction （AMI） and their diagnostic value for AMI.Methods　A total of 176 AMI patients were selected as the experimental group and were divided into ST-segment elevation myocardial infarction （STEMI） group （n=95） and non-ST-segment elevation myocardial infarction （NSTEMI） group （n=81） based on their medical history and electrocardiogram. Meanwhile， 156 patients with negative angiography during the same period were selected as the control group （CON group）. The relative expression levels of lncRNA H19 and miR-22-3p in the serum of the two groups were detected by real-time fluorescence quantitative polymerase chain reaction. The diagnostic value of miR-22-3p and lncRNA H19 in AMI was evaluated by receiver operating characteristic curve （ROC） analysis. The levels of serum cardiac troponin I （cTnI）， creatine kinase （CK）， creatine kinase MB isoenzyme （CK-MB）， and high-sensitivity C-reactive protein （CRP） were detected in all study subjects. Spearman correlation analyses were used to evaluate the correlations between lncRNA H19 and miR-22-3p and cTnI， CK， CK-MB and CRP.Results　Compared with the control group， the expression level of lncRNA H19 was upregulated and the expression level of miR-22-3p was downregulated in the AMI group （all P<0.01）； compared with the NSTEMI group， the expression level of lncRNA H19 was upregulated and the expression level of miR-22-3p was downregulated in the STEMI group （P<0.01）. The areas under the ROC curves （AUC） for the diagnosis of AMI by miR-22-3p， lncRNA H19 and their combination were 0.555， 0.977， and 0.983， respectively. lncRNA H19 was positively correlated with cTnI， CK， CK-MB and CRP （P<0.05）， and negatively correlated with left ventricular ejection fraction （LVEF） （P<0.001）； miR-22-3p was negatively correlated with cTnI， CK and CRP， and positively correlated with LVEF （P<0.05）.Conclusion　The expression level of lncRNA H19 is elevated and the expression level of miR-22-3p decreased in AMI patients， and their levels may have potential value as an auxiliary biomarker for AMI diagnosis and cardiac function assessment.

Abstract:
Objective To investigate the molecular mechanisms related to angiogenesis during the development and progression of diabetic retinopathy（DR）. Methods Angiogenesis-related genes were obtained from the Gen ‑ card website and intersected with differentially expressed genes from DR datasets（GSE60436 and GSE94019）. Functional enrichment and protein-protein interaction（PPI）networks were then used to screen candidate genes and evaluate their diagnostic value. Gene set enrichment analysis（GSEA）was used to explore potential pathways underlying candidate genes，and immune infiltration analysis revealed associations between candidate genes and immune cells. Cellular experiments were conducted to validate the role of fibronectin 1（FN1）in human retinal mi_ crovascular endothelial cells（HRMECs）under high glucose（HG）conditions. Results A total of 237 differen_ tially expressed genes related to angiogenesis were identified，enriched in pathways such as phosphoinositide 3-ki_ nase/protein kinase B signaling pathway（PI3K-Akt）， tumor suppressor protein 53（P53）， tumor necrosis factor （TNF）， and Janus kinase（JAK）/signal transducer and activator of transcription signaling pathway（STAT）. Among them，collagen type I alpha 1 chain（COL1A1）， COL1A2，FN1，TNF，and tumor protein p53（TP53） were key genes with high diagnostic value. GSEA indicated that these genes were involved in multiple signaling pathways，including P53. CIBERSORTx analysis revealed significant associations with the infiltration of multiple immune cells. HG treatment led to the upregulation of FN1. In HG-induced HRMECs，compared with the si-NC control group，si-FN1 significantly reduced cell proliferation，migration，and tube formation，while P53 protein ex_ pression was increased. Conclusion This study reveals the important role of FN1 in angiogenesis in DR and sug_ gests that it may be a potential diagnostic and therapeutic target.

Abstract:
Objective To investigate morphological and functional neural network alterations in the primary visual cortex（V1）during early-stage diabetes. Methods 32 SPF male C57BL/6J mice were randomly divided into con ‑ trol and diabetic groups（n=16）. Type 1 diabetes was induced by intraperitoneal injection of streptozotocin（STZ）， with model validation via oral glucose tolerance test（OGTT）at week 4. Nissl staining was used to assess morpho ‑ logical changes in brain slices of the two groups. Immunohistochemistry was performed to detect the expression of calcium/calmodulin-dependent protein kinase Ⅱ（CaMK Ⅱ ) and somatostatin（SST）in the V1 area. The co- localization density of SST-positive signals on CaMK Ⅱ + neurons was then quantified by ImageJ software. In vivo electrophysiology was used for studying neuronal firing and functional connectivity. Results Compared with the control mice，nissl staining showed diabetic mice exhibited significant V1 thinning（P=0. 02）， reduced neuronal density（P=0. 01）. Immunohistochemistry found decreased SST integral optical density（P=0. 02）and elevated mean fluorescence intensity（P=0. 01），and lower SST density on CaMK Ⅱ + neurons in the diabetic group than in the control group. Compared with the control mice，in vivo electrophysiology studies indicated that diabetic mice showed reduced action potential peak-to-trough ratio and the absolute values of slope（P<0. 01），as well as impaired functional connectivity，including conditional firing，mutual information，and Granger causality（P<0. 000 1）. Con ⁃ clusion Early diabetes induces V1 structural atrophy accompanied by SSTergic system disruption and multi-level functional degradation from single neurons to neural networks.Objective To investigate morphological and functional neural network alterations in the primary visual cortex（V1）during early-stage diabetes. Methods 32 SPF male C57BL/6J mice were randomly divided into con ‑ trol and diabetic groups（n=16）. Type 1 diabetes was induced by intraperitoneal injection of streptozotocin（STZ）， with model validation via oral glucose tolerance test（OGTT）at week 4. Nissl staining was used to assess morpho ‑ logical changes in brain slices of the two groups. Immunohistochemistry was performed to detect the expression of calcium/calmodulin-dependent protein kinase Ⅱ（CaMK Ⅱ ) and somatostatin（SST）in the V1 area. The co- localization density of SST-positive signals on CaMK Ⅱ + neurons was then quantified by ImageJ software. In vivo electrophysiology was used for studying neuronal firing and functional connectivity. Results Compared with the control mice，nissl staining showed diabetic mice exhibited significant V1 thinning（P=0. 02）， reduced neuronal density（P=0. 01）. Immunohistochemistry found decreased SST integral optical density（P=0. 02）and elevated mean fluorescence intensity（P=0. 01），and lower SST density on CaMK Ⅱ + neurons in the diabetic group than in the control group. Compared with the control mice，in vivo electrophysiology studies indicated that diabetic mice showed reduced action potential peak-to-trough ratio and the absolute values of slope（P<0. 01），as well as impaired functional connectivity，including conditional firing，mutual information，and Granger causality（P<0. 000 1）. Con ⁃ clusion Early diabetes induces V1 structural atrophy accompanied by SSTergic system disruption and multi-level functional degradation from single neurons to neural networks.

Abstract:
Objective To investigate the short-term clinical outcomes and long-term prognostic value of preserving the superior rectal artery（SRA）versus high ligation of the inferior mesenteric artery（IMA）in laparoscopic sig‑ moid colon cancer resection（LSCER）for sigmoid colon cancer（SCC）. Methods A retrospective comparative analysis was performed of 316 SCC patients who underwent LSCER. Patients were divided into the SRA preserva ‑ tion group（Group A，132 cases）and IMA high ligation group（Group B，184 cases）according to the vascular man ‑ agement strategy. Short-term clinical indicators and long-term follow-up outcomes were compared between the two groups. Results There were no significant differences in baseline data between the two groups. The mean opera‑ tion time of Group A was（190. 0 ± 61. 9）min，which was significantly longer than that of Group B［（165. 0 ± 49. 1）min，P<0. 001］. However，Group A demonstrated significantly shorter time to drain removal，time to first flatus，and postoperative hospital stay（P<0. 001）. No significant differences were observed in intraoperative blood loss，number of lymph nodes dissected，or incidence of anastomotic leakage（AL）between the two groups. Long- term follow-up results showed that there were no significant differences in 3-year overall survival rate and 3-year disease-free survival rate between the two groups. Conclusion Laparoscopic sigmoid colon cancer resection with SRA preservation is safe and feasible for the treatment of SCC. Although the operation time is slightly prolonged，it can effectively promote short-term postoperative recovery without affecting the curative effect of tumor resection or long-term prognosis.

Abstract:
Objective To investigate the effects of luteinizing hormone（LH）levels on in vitro fertilization and em ‑ bryo transfer（IVF-ET）outcomes in an infertile population with normal ovarian reserve（NOR）undergoing con‑ trolled ovarian hyperstimulation（COH）with gonadotropin-releasing hormone antagonist protocol，and to explore the reasonable level intervals of LH during the follicular phase. Methods A retrospective analysis was conducted on 911 NOR infertility patients undergoing IVF-ET treated with gonadotropin-releasing hormone antagonist proto ‑ col. Patients were grouped based on baseline LH（bLH）levels，LH levels on the day of antagonist addition，and LH levels on the day of human chorionic gonadotropin（hCG）injection. According to bLH levels，patients were di‑ vided into Group A2（n=391）and Group B2（n=391）. Based on LH levels on the day of antagonist addition，pa‑ tients were divided into Group A3（n=600）and Group B3（n=311）. According to LH levels on the day of hCG in ‑ jection，patients were classified into Group A4（n=225），Group B4（n=227），Group C4（n=231），and Group D4 （n=228）. The clinical characteristics，COH outcomes，and pregnancy outcomes of patients in each group were compared. Results ① The bLH level could be used as a predictor of assisted reproduction outcomes in an infertile population with NOR，and going for assisted reproduction at bLH≥4. 51 IU/L was more favorable for the develop ‑ ment of pregnancy outcomes；② Addition of antagonist at LH≥5 IU/L on the day of antagonist addition improved pregnancy outcome ；③ On the day of hCG injection，there was no statistically significant difference in pregnancy outcomes among the four groups ；④ Analyses using generalized linear models revealed that basal follicle- stimulating hormone（bFSH）， basal estradiol（bE2）， Gn days，Gn volume and 2PN affected the number of good quality embryos. Conclusion In an infertile population with NOR using gonadotropin-releasing hormone antago‑ nist，bLH and gonadotropin-releasing hormone antagonist addition day LH levels may influence IVF outcomes. LH levels should be controlled at 5 IU/L and above on antagonist addition days，and bLH levels should be controlled at 4. 51 IU/L and above.

Abstract:
Objective To explore the risk factors and alterations in functional connectivity of brain networks in end - stage renal disease（ESRD）patients complicated by frailty. Methods A total of 84 patients with ESRD were enrolled and divided into the frailty group（31 cases）and non-frailty group（53 cases）according to the Chinese version of the Til‑ burg Frailty Indicator（TFI）. Cranial magnetic resonance imaging（MRI）data，neuropsychological test results and clinical data were collected for all subjects. Logistic regression analysis was used to explore risk factors for frailty in ESRD patients，and image preprocessing and independent component analysis were performed to compare differences in brain network functional connectivity between the two groups. Results Logistic regression analysis showed that high blood urea nitrogen level，depression，malnutrition，and sleep disturbance were risk factors for frailty in patients with ESRD（P<0. 05），while a higher level of education served as a protective factor（P<0. 05）. Compared with the non- frailty group，the frailty group exhibited significantly weakened functional connectivity between the sensorimotor net ‑ work and the visual network（P<0. 05）. Conclusion High levels of blood urea nitrogen，depression，malnutrition，and sleep disorders can increase the risk of frailty in patients with end-stage renal disease（ESRD），while a higher educa‑ tional level may reduce this risk. Patients with ESRD complicated by frailty exhibit a significant decrease in functional connectivity between the sensorimotor network（SMN）and visual network（VN），which provides novel imaging evi‑ dence for exploring the central neural mechanisms underlying the development of frailty in ESRD patients.

Abstract:
Objective To explore the effects of vascular endothelial growth factor A（VEGFA）rs2010963 and rs3025039 polymorphisms on chemotherapy toxicities and clinical prognosis in children with brain tumors. Meth ⁃ ods A total of 104 pediatric patients with brain tumors receiving standardized chemotherapy were enrolled. Matrix-assisted laser desorption/ionization time of flight mass spectrometry was used for VEGFA rs2010963 and rs3025039 genotyping. The χ² test was applied to analyze the association between genotypes and chemotherapy‑related toxicities. Cox regression was used to evaluate the correlations of clinicopathological charac ‑ teristics and genotypes with the progression‑free survival（PFS）. In addition，bioinformatic analyses were con ‑ ducted to investigate the regulatory factors potentially affected by the two SNP loci. Results The VEGFA expres‑ sion in brain tumors（5. 17± 1. 81）was significantly higher than that in normal tissues（4. 33± 1. 56，P<0. 001）. Patients with high VEGFA expression had significantly worse overall survival than patients with low VEGFA expres‑ sion（P<0. 001）. Among the 104 children with brain tumors included，the rs2010963 CC，CG，and GG genotypes accounted for 14. 42%，55. 77%，and 29. 81%，respectively. The frequencies of C and G alleles were 42. 31% and 57. 69%，respectively. The rs3025039 CC，CT，and TT genotypes accounted for 70. 19%，25. 96%，and 3. 85%，respectively. The frequencies of C and T alleles were 83. 17% and 16. 83%，respectively. The children with the rs3025039 CC genotype had significantly higher incidences of thrombocytopenia（46. 58%）and gastroin‑ testinal toxicity（56. 16%）than CT genotype carriers（22. 22% and 33. 33%，respectively，P<0. 05），and signifi‑ cantly lower incidences of coagulation disorders（4. 11%）than TT genotype carriers（50. 00%，P<0. 05）. The inci‑ dence of hyperlipidemia（2. 74%）was significantly lower than that in the CT genotype carriers（14. 82%，P< 0. 05）. The tumor type and the rs2010963 genotype were significantly associated with PFS（P<0. 05）in univari‑ able and multivariable Cox regression analysis. Bioinformatic analysis indicated that the rs2010963 and rs3025039 polymorphisms regulated VEGFA expression by affecting the binding of transcription factors and miRNAs to their target gene sequences，respectively. Conclusion The VEGFA rs3025039 CC genotype is a risk factor for thrombo ‑ cytopenia and gastrointestinal toxicity，and a protective factor for coagulation disorders and hyperlipidemia. The rs2010963 CG genotype is a protective factor for brain tumor progression.

Abstract:
Objective To investigate the differences in pregnancy outcomes between dichorionic diamniotic （DCDA）twin pregnancies conceived via assisted reproductive technology（ART）and those from spontaneous con‑ ception，in order to provide evidence for the perinatal management of ART-conceived DCDA twins. Methods A retrospective analysis was conducted on the clinical data of 549 DCDA twin parturients. Based on the mode of con‑ ception，they were divided into ART group（423 cases）and spontaneous conception group（126 cases）. The base‑ line characteristics，pregnancy complications and neonatal outcomes were compared between the two groups. Re ⁃ sults Regarding baseline characteristics，the ART group had higher maternal age，number of previous abortions， proportions of primiparas，proportions of advanced maternal age and greater 24-hour postpartum blood loss，but lower parity and proportion of scarred uterus compared to the spontaneous conception group（P<0. 05）. For preg‑ nancy complications，the ART group exhibited higher incidences of hypertensive disorders of pregnancy，placental adhesion，postpartum hemorrhage and a higher application rate of uterine artery ascending branch ligation（P< 0. 05）. There were no statistically significant differences in neonatal-related indicators between the two groups. Af‑ ter adjusting for confounding factors such as age，number of abortions and parities，ART remained an independent risk factor for hypertensive disorders of pregnancy，placental adhesion and postpartum hemorrhage（P<0. 05）， while it did not increase the risk of adverse neonatal outcomes. Further subgroup analysis by gestational age showed that in the late preterm subgroup，the risk of ART-related pregnancy complications remained elevated（P< 0. 05）. Additionally，neonatal outcomes showed no significant difference between the two groups in both the late preterm and term subgroups. Conclusion ART increases the risk of hypertensive disorders of pregnancy，placen‑ tal adhesion and postpartum hemorrhage in DCDA twin parturients. Clinically，enhanced monitoring of blood pres‑ sure during pregnancy and blood loss during delivery and postpartum，along with targeted prevention and manage ‑ ment of these complications，are crucial for improving pregnancy outcomes.

Abstract:
Objective To explore the incidence of major low anterior resection syndrome（LARS）within six months after sphincter-preserving surgery for rectal cancer and to analyze its associated factors. Methods Clinical records，postoperative rehabilitation data and LARS scores were collected from 889 patients at 3 months postopera_ tively and from 844 patients at 6 months postoperatively. Patients were divided into a major LARS group and a non- major LARS group. Multivariable Logistic regression was used to analyze factors associated with major LARS at 3 and 6 months postoperatively，and the predictive value of the models was assessed using the Hosmer-Lemeshow goodness-of-fit test and the receiver operating characteristic （ROC）. Results Among patients undergoing sphincter-preserving surgery for rectal cancer，247 patients（27. 8%）had major LARS at 3 months postoperatively and 181 patients（21. 4%）at 6 months postoperatively. Multivariable Logistic regression showed that Kegel exer_ cises，preoperative radiotherapy，anastomotic leakage，a tumor distance from the anal verge of ≤5 cm，and an anas_ tomotic distance from the anal verge of ≤5 cm were independent factors for major LARS at 3 months postoperatively （P<0. 05）. At 6 months postoperatively，occasional Kegel exercises during 0~3 months，Kegel exercises during 4~ 6 months，hydrotherapy during 4~6 months，preoperative chemotherapy，a tumor distance from the anal verge of ≤ 5 cm，and an anastomotic distance from the anal verge of ≤5 cm were independent factors（P<0. 05）. The models at 3 and 6 months postoperatively showed goodness-of-fit test P values of 0. 986 and 0. 517 and area under the curve（AUCs）of 0. 843 and 0. 870，indicating good predictive value. Conclusion The incidence of major LARS within six months after sphincter-preserving surgery for rectal cancer is relatively high，and greater attention should be paid to patients who receive preoperative chemoradiotherapy，develop anastomotic leakage，or have tumor and anastomotic distances from the anal verge of ≤5 cm. Patients should also be advised that early postoperative Kegel exercises and hydrotherapy may reduce the incidence of major LARS.

Abstract:
Objective　To analyze the effects of the progestin-primed ovarian stimulation （PPOS） protocol on embryo euploidy rate and clinical pregnancy outcomes in infertile patients.Methods　Women who underwent Preimplantation genetic testing for aneuploidy （PGT-A） cycles were selected as study participants （a total of 656 cycles and 3 081 blastocysts）. Participants were divided into two age groups （≤35 years and >35 years） and further stratified by ovarian stimulation protocol： the Progestin-Primed Ovarian Stimulation （PPOS） group （n=48 and 60， respectively）， the Luteal Phase Long Protocol （LP） group （n=160 and 57， respectively）， and the Gonadotropin-Releasing Hormone Antagonist Protocol （AP） group （n=220 and 111， respectively）. Baseline characteristics， ovarian stimulation outcomes， and embryo development parameters were compared among the three protocols within each age group. Additionally， clinical pregnancy outcomes of the first frozen embryo transfer （FET） cycle were compared. The primary outcome measure was the embryo euploidy rate.Results　① In the ≤35 years age group， baseline follicle-stimulating hormone （bFSH） levels were significantly higher in the PPOS group compared to the LP and AP groups， and the bFSH/bLH ratio was significantly higher in the PPOS group than in the AP group （P < 0.05）. In the >35 years age group， bFSH levels in both the PPOS and AP groups were significantly higher than in the LP group （P < 0.05）.② In the ≤35 years group， there were no significant differences in the rates of metaphase II （MII） oocytes， 2-pronuclei （2 PN） zygotes， cleavage， 2 PN cleavage， blastocyst formation， high-quality embryos， or embryo euploidy between the PPOS group and either the LP or AP groups. However， the PPOS group showed significantly lower values for the following parameters compared to the other two groups： total oocytes retrieved， number of MII oocytes， number of 2 PN zygotes， number of cleaved embryos， number of 2 PN cleaved embryos， number of blastocysts formed， number of high-quality embryos， number of high-grade blastocysts， number of transferable embryos， number of cryopreserved embryos， number of biopsied blastocysts， number of euploid embryos， and the rate of obtaining at least one euploid embryo. In the >35 years group， no significant differences were observed among the PPOS， LP， and AP groups in the rates of MII oocytes， 2 PN cleavage， blastocyst formation， or in the number of euploid embryos， euploidy rate， and the rate of obtaining at least one euploid embryo. However， the PPOS group had significantly lower numbers of total oocytes retrieved， MII oocytes， 2 PN zygotes， cleaved embryos， 2 PN cleaved embryos， blastocysts formed， transferable embryos， and cryopreserved embryos compared to the LP group （P<0.05）. The high-quality embryo rate was significantly lower in the PPOS group than in the AP group （P<0.05）. The numbers of high-quality embryos， high-grade blastocysts， and biopsied blastocysts were significantly lower in the PPOS group than in both the LP and AP groups （P<0.05）. Notably， the 2 PN fertilization rate was significantly higher in the PPOS group than in the AP group， and the cleavage rate was significantly higher in the PPOS group than in both the LP and AP groups （P<0.05）.③ No significant differences were found among the three groups in the biochemical pregnancy rate，clinical pregnancy rate，live birth rate， early miscarriage rate，and late miscarriage rate following the first FET cycle. Conclusion Compared to the other two protocols，the PPOS protocol does not significantly affect embryo euploidy or clinical pregnancy rates. How_ ever，in freeze-all cycles with PGT-A，the PPOS protocol does not reduce the rate of chromosomally normal em_ bryos but may decrease the total number of available embryos. It may not be the optimal choice for younger women， but it can be considered as a viable option for women of advanced maternal age.

Abstract:
Objective To investigate the association between endothelin receptor type A gene（EDNRA）and con‑ genital bilateral absence of the vas deferens（CBAVD）. Methods This case-control study consisted of 124 sub‑ jects with CBAVD and 100 fertile controls. Two single nucleotide polymorphisms（SNPs：rs5335 and rs1801708） in the EDNRA gene were genotyped via PCR，PCR-RFLP analysis，and direct sequencing. Results No signifi‑ cant differences existed between EDNRA polymorphisms and CBAVD phenotype（rs1801708：P=0. 220 2，0. 163 2； rs5335：P=0. 805 8，0. 818 6）. However，the rs1801708-rs5335 haplotype AG was notably associated with an in ‑ creased risk of CBAVD（P=0. 008 6，OR=2. 178，95% CI：1. 207-3. 929）. A significantly protective effect of rs1801708-rs5335 haplotype GG on CBAVD（P=0. 038 5，OR= 0. 671，95% CI：0. 460-0. 980）was observed. After Bonferroni correction，the result for haplotype A-G remained significant（P =0. 008 6<0. 012 5），while hap‑ lotype G-G was not significant. Conclusion The rs1801708-rs5335 AG haplotype of EDNRA is a potential risk fac‑ tor for CBAVD development in Han Chinese.

Abstract:
Objective To explore the value of deep learning reconstruction（DLR）in liver magnetic resonance im ‑ aging（MRI）by comparing the single-shot half-fourier rapid spin-echo sequence with DLR（HASTEDLR）and diffusion-weighted imaging sequence with DLR（DWIDLR）against the conventional BLADE and conventional DWI sequences. Methods 70 patients underwent MRI examinations. Two observers independently evaluated the image quality of each sequence（including liver edge，blood vessels，lesion clarity，etc. ）. Additionally，quantitative evaluation was conducted by measuring the signal to noise ratio（SNR）of liver parenchyma and lesions，contrast to noise ratio（CNR）of lesions，as well as apparent diffusion coefficient（ADC）values from conventional DWI and DWIDLR. Intraclass correlation coefficient（ICC）was used to evaluate the consistency between the two observers. Results The inter-observer consistency was high（ICC：0. 84-0. 97）. The scanning time was reduced by 92. 63% for HASTEDLR and 50% for DWIDLR sequences，respectively. The lesion clarity score of the HASTEDLR group was sig‑ nificantly better than that of the BLADE group（P<0. 001）， with artifacts reduced in both DLR sequences（P< 0. 05）. The HASTEDLR group demonstrated higher SNR and CNR，while the DWIDLR group showed higher SNR and ADC values（all P<0. 05）. Conclusion The DLR technology can enhance the efficiency of liver MRI scans，im‑ prove image quality，and reduce artifacts，demonstrating promising application prospects.

Abstract:
Objective To evaluate the application effect of morphology driven preparation technique in indirect restoration of posterior teeth. Methods 84 patients with dental defects were selected and divided into the control group and the experimental group randomly，with 42 patients in each group（a total of 84 teeth）. Traditional prepa_ ration and morphology driven preparation techniques were used to complete dental preparation and adhesive glass ceramic restorations，respectively. The satisfaction，masticatory function，World Dental Federation（FDI）Scores for edge fracture and fixation，edge adaptability index，periodontal bleeding index（BI）and plaque index（PLI）of the two groups of patients were evaluated and various indicator data were recorded and statistically analyzed at 3 and 6 months after restoration completion. Results There was no statistically significant difference in satisfaction between the two groups after 3 and 6 months of repair；Both groups of data showed a significant improvement in masticatory function. There was no statistical difference after 3 and 6 months of repair. However，a statistically sig_ nificant difference in masticatory function was noted after 6 months of repair compared with before and after 3 months of repair. As the repair time increased，the masticatory effect improved significantly. There was no statisti_ cally significant difference in the edge fracture and fixation index between the two groups at 3 months after repair， but there was a statistically significant difference between the two groups at 6 months with the experimental group outperforming the control group（P<0. 05）. The edge adaptability，BI，and PLI index were statistically significant between the two groups after 3 and 6 months of repair and the experimental group had a significant advantage（P< 0. 05）. Conclusion Both types of tooth preparation techniques can improve patient masticatory function and reach high satisfaction after repair. However，morphology driven preparation technique has significant advantages in edge adaptability，edge fracture and fixation index scores and improve periodontal conditions. This technique is an effective method for improving dental preparation in adhesive indirect restoration of posterior teeth.

Abstract:
Cornelia de Lange syndrome（CdLS）is a rare genetic disorder characterized by multisystem develop ‑ mental abnormalities，with its core pathogenic mechanism closely linked to dysfunction of the cohesin complex. In‑ tegrated multi‑omics evidence revealed that cohesin dysfunction disrupted three‑dimensional chromatin architecture and epigenetic homeostasis，triggered genome‑wide transcriptional dysregulation，and perturbed the regulatory net‑ works of signaling pathways such as Wnt/β‑catenin，TGF‑β/BMP，and Sonic Hedgehog（SHH）. These distur‑ bances collectively drove multisystem phenotypes involving the neurological，cardiovascular and skeletal systems， and laid a theoretical foundation for further understanding of the pathological mechanisms of CdLS and optimizing molecular diagnosis. This review summarized the genetic basis（including epigenetic dysregulation mechanisms） and disruption of key developmental signaling pathways in CdLS，and discussed strategies for optimizing molecular diagnosis.

Abstract:
Rheumatoid arthritis（RA）is an autoimmune disease characterized by chronic synovial inflammation and joint destruction，with a complex pathogenesis. Macrophage polarization plays a pivotal role in disease progres_ sion. Recent studies demonstrate that microRNA（miRNA），as critical post-transcriptional regulators，can partici_ pate in the inflammatory response process by regulating the polarization state of macrophages，thereby aggravating RA joint damage. This review summarizes the mechanism of action of various miRNA in regulating macrophage po_ larization，and emphasizes their potential therapeutic value in RA.

Abstract:
Cranial bone defect repair remains a significant challenge in craniomaxillofacial surgery，particularly when defects exceed the critical size defect（CSD）threshold，rendering them incapable of spontaneous healing without external intervention. Mechanical forces—including compression，tension，and shear stress—play a piv_ otal role in cranial development and regeneration. These forces regulate osteoblast differentiation and bone regen_ eration by activating key mechanotransduction pathways，such as the Wnt/β-catenin signaling pathway，the piezo_ electric mechanosensitive ion channel Piezo1，and the transcriptional co-activators Yes-associated protein（Yes- Associated Protein，YAP）and Transcriptional co-activator with PDZ-binding motif（Transcriptional co-activator with PDZ-binding motif，TAZ）. Distinct types of mechanical forces exert specific effects on cellular behavior and the microenvironment. Clinically，applications such as Negative Pressure Wound Therapy（NPWT）have demon_ strated efficacy in promoting angiogenesis-osteogenesis coupling，while tensile forces stimulate the dura mater to se_ crete osteogenic factors. Preliminary studies using artificial periosteum and other biomaterials have further vali_ dated that appropriate mechanical stimulation enhances bone repair. This review summarizes the effects of various mechanical forces and their associated signaling pathways on osteoblasts and their microenvironment，alongside an overview of current technological applications in this field.