Objective To explore the mechanism by which zinc-regulated transporters, iron-regulated transporter-likeprotein 4 (ZIP4) regulates glycolysis and its impact on tumor progression in cholangiocarcinoma (CCA), providing a theoretical basis for targeted therapy of CCA. Methods ZIP4 expression in CCA was analyzed using the GEPIA database. Immuno-histochemistry (IHC) was used to detect ZIP4 expression in 20 paired CCA and adjacent non-tumor tissues. Stable ZIP4-overexpressing CCA cell lines (ZIP4-OE) were established. Gene set enrichment analysis was used to screen differentially expressed genes and pathways in ZIP-OE CCA cells. ZIP4, N-myc proto-oncogene protein (MYCN), and histone-lysine N-methyltransferase 2E (KMT2E) were knocked down using small interfering RNAs (siRNAs). The expression of glycolysis-related gene (glucose transporter 1 [Glut1], hexokinase 2 [HK2], and lactate dehydrogenase A [LDHA]) was measured by real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR). Glycolytic activity was assessed by measuring the extracellular acidification rate (ECAR). Cell proliferation was evaluated using colony formation assays, and cell migration was assessed using Transwell assays. A xenograft mouse model was constructed to examine CCA tumor growth. Protein levels of ZIP4, KMT2E, H3K4me3 (tri-methylation of lysine 4 on histone H3), and MYCN were detected by Western blotting. Results GEPIA database analysis and IHC results confirmed significantly higher ZIP4 expression levels in CCA tissues compared to adjacent non-tumor tissues (P＜0.05). Compared to the control group, the ZIP4-OE group exhibited a significantly increased ECAR, along with significantly enhanced proliferation and migration abilities (P＜0.01). Conversely, knockdown of ZIP4 suppressed CCA cells proliferation and migration. GEPIA analysis indicated that ZIP4 upregulates the transcription of oncogene MYCN, as well as glycolysis-related genes. Knockdown of MYCN abolished the ZIP4 overexpression-induced upregulation of Glut1, HK2, and LDHA gene transcription, reduced glycolysis, and significantly inhibited CCA cell proliferation and migration (P＜0.05). Mechanistic studies demonstrated that ZIP4 increases H3K4me3 level via KMT2E, leading to MYCN transcription. Knockdown of KMT2E in CCA cells suppressed the ZIP4 overexpression-induced enhancement in H3K4me3 modification, resulting in MYCN downregulation and significantly reduced CCA cells proliferation and migration (P＜0.05). Conclusions ZIP4 upregulates H3K4me3 modification through KMT2E, which recruits transcription factors to activate the transcription of MYCN. This subsequently enhances cellular glycolysis and promotes the proliferation and migration of CCA cells.

Objectives To retrospectively analyze the clinical outcomes of lateral malleolus osteotomy combined with cannulated screws and inverted proximal humerus locking plate fixation in the treatment of end-stage ankle osteoarthritis.Methods A clinical study was conducted on 21 patients with end-stage ankle osteoarthritis who were treated at our hospital from June 2018 to May 2022.Preoperative evaluations included weight-bearing X-rays to assess lower limb alignment and joint space,CT to evaluate the degree of bone sclerosis,and MRI to examine the extent of articular cartilage damage.The surgery involved a lateral malleolus osteotomy approach,using the resected lateral malleolus cartilage for grafting at the fusion site,and fixation of the tibiotalar joint with cannulated screws and an inverted proximal humerus locking plate.After the operation,the joint fusion was evaluated by X-ray films followed up in the outpatient department.The clinical efficacy was evaluated by using the Visual Analogue Scale(VAS)for pain and the Ankle and hindfoot Function Score of the American Orthopaedic Foot and Ankle Society(AOFAS).Results Twenty patients were followed up(95.2％follow-up rate)over a period of 14 to 40 months,with an average follow-up duration of 27.8 months.All 21 patients experienced primary wound healing.One patient developed lateral foot numbness,and 19 patients achieved primary osseous fusion.One patient with diabetes required secondary bone grafting with PRP injection to achieve fusion.Postoperative AOFAS scores significantly improved from a preoperative average of 45.7±3.3 to 84.7±3.3,while VAS scores for pain decreased from 7.54±1.83 preoperatively to 1.94±0.33 postoperatively(P＜0.05).Conclusion Lateral malleolus osteotomy combined with cannulated screws and inverted proximal humerus locking plate fixation for the treatment of end-stage ankle osteoarthritis demonstrated a high rate of fusion and low complication rates,The short-term outcomes are favorable,and the procedure eliminates the need for iliac bone grafting,indicating its potential for broader clinical application.

Hypoxic-ischemic (HI) brain damage poses a high risk of death or lifelong disability, yet effective treatments remain elusive. Here, we demonstrated that miR-100-5p levels in the lesioned cortex increased after HI insult in neonatal mice. Knockdown of miR-100-5p expression in the brain attenuated brain injury and promoted functional recovery, through inhibiting the cleaved-caspase-3 level, microglia activation, and the release of proinflammation cytokines following HI injury. Engineered extracellular vesicles (EVs) containing neuron-targeting rabies virus glycoprotein (RVG) and miR-100-5p antagonists (RVG-EVs-Antagomir) selectively targeted brain lesions and reduced miR-100-5p levels after intranasal delivery. Both pre- and post-HI administration showed therapeutic benefits. Mechanistically, we identified protein phosphatase 3 catalytic subunit alpha (Ppp3ca) as a novel candidate target gene of miR-100-5p, inhibiting c-Fos expression and neuronal apoptosis following HI insult. In conclusion, our non-invasive method using engineered EVs to deliver miR-100-5p antagomirs to the brain significantly improves functional recovery after HI injury by targeting Ppp3ca to suppress neuronal apoptosis.
Animals ; MicroRNAs/metabolism* ; Extracellular Vesicles/metabolism* ; Mice ; Recovery of Function/physiology* ; Hypoxia-Ischemia, Brain/therapy* ; Mice, Inbred C57BL ; Antagomirs/administration & dosage* ; Male ; Animals, Newborn ; Apoptosis/drug effects* ; Brain Injuries/metabolism* ; Glycoproteins ; Peptide Fragments ; Viral Proteins

Fractional flow reserve (FFR), an established modality for functionally assessing coronary artery disease, is increasingly applied to diagnose and manage lower extremity arterial disease. By incorporating functional parameters, FFR enhances revascularization precision by quantifying the hemodynamic impact of stenotic lesions, thereby overcoming limitations of conventional imaging. Key clinical applications in lower extremity disease include functional assessment in moderate intermittent claudication, post-vascular preparation strategy optimization, and predicting revascularization outcomes and complications. Advances in pressure wire and microcatheter systems, alongside non-invasive imaging-derived FFR techniques, are improving its feasibility and applicability. However, widespread adoption is challenged by the complex anatomy of the lower extremity arterial system, frequent severe calcification and diffuse disease, and a current lack of standardized FFR cutoff values. Promoting the standardized use of FFR is crucial for shifting the clinical management paradigm from anatomy-based repair toward functional reconstruction.
Humans ; Lower Extremity/blood supply* ; Peripheral Arterial Disease/diagnosis* ; Fractional Flow Reserve, Myocardial ; Endovascular Procedures/methods* ; Intermittent Claudication/physiopathology*

Objectives To retrospectively analyze the clinical outcomes of lateral malleolus osteotomy combined with cannulated screws and inverted proximal humerus locking plate fixation in the treatment of end-stage ankle osteoarthritis.Methods A clinical study was conducted on 21 patients with end-stage ankle osteoarthritis who were treated at our hospital from June 2018 to May 2022.Preoperative evaluations included weight-bearing X-rays to assess lower limb alignment and joint space,CT to evaluate the degree of bone sclerosis,and MRI to examine the extent of articular cartilage damage.The surgery involved a lateral malleolus osteotomy approach,using the resected lateral malleolus cartilage for grafting at the fusion site,and fixation of the tibiotalar joint with cannulated screws and an inverted proximal humerus locking plate.After the operation,the joint fusion was evaluated by X-ray films followed up in the outpatient department.The clinical efficacy was evaluated by using the Visual Analogue Scale(VAS)for pain and the Ankle and hindfoot Function Score of the American Orthopaedic Foot and Ankle Society(AOFAS).Results Twenty patients were followed up(95.2％follow-up rate)over a period of 14 to 40 months,with an average follow-up duration of 27.8 months.All 21 patients experienced primary wound healing.One patient developed lateral foot numbness,and 19 patients achieved primary osseous fusion.One patient with diabetes required secondary bone grafting with PRP injection to achieve fusion.Postoperative AOFAS scores significantly improved from a preoperative average of 45.7±3.3 to 84.7±3.3,while VAS scores for pain decreased from 7.54±1.83 preoperatively to 1.94±0.33 postoperatively(P＜0.05).Conclusion Lateral malleolus osteotomy combined with cannulated screws and inverted proximal humerus locking plate fixation for the treatment of end-stage ankle osteoarthritis demonstrated a high rate of fusion and low complication rates,The short-term outcomes are favorable,and the procedure eliminates the need for iliac bone grafting,indicating its potential for broader clinical application.

Objective:To analyze the biological characteristics, phylogenic features and clinical significance of SQ219 and SQ220 isolated from clinical sputum and midstream urine specimens.Methods:The culture and biochemical characteristics of the two strains were observed. VITEK2 System, drug sensitivity testing and MALDI-TOF mass spectrometry were used for bacterial identification. Phylogenetic analysis based on 16S rRNA and core genome was performed. The average nucleotide identity (ANI) based on whole genome sequences was calculated.Results:SQ219 and SQ220 were Gram-stain-negative, aerobic, catalase- and oxidase-positive, and non-motile bacteria. Their optimum growth was observed in NaCl-free medium at 30℃ and pH7. Flexirubin-type pigments were produced by SQ220 on Colombia blood agar, but not by SQ219. Both SQ219 and SQ220 were resistant to aztreonam, amikacin, tobramycin and colistin, which was consistent with the drug resistance phenotype of genus Chryseobacterium. The genome sequences of SQ219 and SQ220 were 5.08 Mb and 4.80 Mb in length, and the G+ C contents were 36.72% and 36.36%, respectively. Both strains carried β-lactam resistance gene ( blaCGA). 16S rRNA phylogenetic analysis showed that SQ219 and SQ220 were closely related to Chryseobacterium gambrini DSM18014 T with the similarities of 98.93% and 98.36%, respectively. Core genome phylogenetic analysis revealed that SQ219 and SQ220 were highly homologous to Chryseobacterium gambrini DSM18014 T. However, the ANI values between the two strains and Chryseobacterium gambrini DSM18014 T were 92.49% and 93.27%, respectively, below the threshold for prokaryotic species identification. Conclusions:Based on the phenotypic and phylogenetic data, SQ219 and SQ220 represent a novel species of the genus Chryseobacterium. This study would help promote the understanding of the evolution of Chrysobacterium and provide reference for the identification of new species of Chrysobacterium.

Objective: To investigate the oxidative stress injury of nano zinc oxide nanoparticles(ZnO NPs) on human myocardial cells (AC16) ，and to analyze the mechanism of ZnO NPs from the transcriptome level． Methods: Dynamic light scattering (DLS) was used to characterize and detect ZnO NPs．After AC16 cells were exposed to ZnO NPs at different doses and at different times，the cell survival rate was determined by CCK-8 method．AC16 cells were divided into control group，ZnO NPs (50，100，200 μmol /L) ，after 6 h treatment，the mitochondrial membrane potential (MMP) and reactive oxygen species (ROS) were measured．AC16 cells were divided into control group，50 μmol /L ZnO NPs group and 200 μmol /L ZnO NPs group．After 6 h exposure，total RNA was extracted by TRIzol for transcriptome analysis ，and the differentially expressed genes were enriched by gene body ( GO) ，Kyoto Encyclopedia of Genes and Genomes (KEGG) . Results : The results of DLS showed that the hydrodynamic diameter was ( 192. 2 ± 1. 63 ) nm and the Zeta potential was ( －23. 26 ± 1. 05 ) mV． CCK-8 results showed that the survival rate of AC16 cells decreased with the increase of dose and time of exposure to ZnO NPs. Fluorescence quantification showed that with the increase of ZnO NPs exposure dose，MMP significantly decreased at 100 μmol /L ZnO NPs(P＜0. 05) ，and ROS significantly increased at 50 μmol /L ZnO NPs(P＜0. 05) ．Using the multifunctional microplate reader，it was observed that MMP and ROS were statistically significant at 100 and 50 μmol /L ZnO NPs，respectively，showing a decrease in MMP and an increase in ROS．Transcriptome analysis showed that 1 071 genes were enriched in the 50 μmol /L ZnO NPs group compared with the control group，including 561 up-regulated genes and 510 down-regulated genes．Compared with the control group，7 164 genes were enriched in 200 μmol /L ZnO NPs group，including 4 098 up-regulated genes and 3 066 down-regulated genes．GO and KEGG analysis showed that the differential genes were mainly concentrated in ROS，antioxidant activity，mitochondrial cytochrome C release，apoptosis and other signaling pathways． Conclusion ZnO NPs can decrease the survival rate of AC16 cells and induce mitochondrial damage and oxidative stress，among which ROS-mediated oxi- dative stress and mitochondrial function changes are important toxic mechanisms of ZnO NPs induced AC16 cytotoxicity.

Objective:To analyze the application of thinking visual guidance teaching model in anesthesia clinical practice teaching.Methods:Taking the implementation time (March 2019) of thinking visual guidance teaching model in our hospital as the limit, 56 interns who came to our hospital for anesthesia practice before the implementation were included as control group, and 61 interns enrolled after the implementation (from March 2019 to March 2020) were included in study group. The mastery status of professional skills (duration of anesthesia operation, success rate of one-time anesthesia puncture and success rate of one - time tracheal intubation), professional knowledge assessment results and professional attitude changes were compared between the two groups at 2 months and 6 months of training. Self-evaluation after 6 months of training was compared between the two groups. SPSS 18.0 was used for t test and chi-square test. Results:After 6 months of training, the mastery status of professional skills (duration of anesthesia operation, success rate of one-time anesthesia puncture, and success rate of one-time tracheal intubation) of the two groups of interns were significantly improved compared with those after 2 months of training, and the above indicators of study group were significantly better than those of control group (all P<0.05). After 6 months of training, the scores of assessment results of professional knowledge (basic knowledge, understanding and memory, case analysis, emergency handling) and professional attitude (behavior, language, initiative and adaptability) in the two groups of interns were significantly higher than those after 2 months of training, and the above indicators scores of study group were significantly higher than those of control group (all P<0.05). The scores of self-evaluation (learning efficiency, learning atmosphere, learning ability, self-confidence and satisfaction) of study group at 6 months of training were significantly higher than those of control group at the same period (all P<0.05). Conclusion:Thinking visual guidance teaching model can effectively improve the professional knowledge and operation skills of anesthesia clinical interns, and it has a good application effect.

Rheumatoid arthritis (RA) is an autoimmune disease and is mainly characterized by abnormal proliferation of fibroblast-like synoviocytes (FLS). The up-regulated cellular membrane expression of G protein coupled receptor kinase 2 (GRK2) of FLS plays a critical role in RA progression, the increase of GRK2 translocation activity promotes dysfunctional prostaglandin E4 receptor (EP4) signaling and FLS abnormal proliferation. Recently, although our group found that paeoniflorin-6'-

Objective To study the use of preoperative indocyanine green retention test at 15 minutes (ICG R15) in the prediction of liver functional reserve in patients with hilar cholangiocarcinoma (HCCA).Methods The clinical data of 62 patients with HCCA treated in our department from March 2016 to March 2018 was reviewed.The relationship between preoperative ICG R15 and postoperative hepatic insufficiency was analyzed.The relationship between preoperative ICG R15 and Child-Pugh scoring was also studied.Univariate analysis was used to evaluate the risk factors of postoperative liver dysfunction.Logistic regression was used to assess the independent risk factors of postoperative liver dysfunction.The regression equation between independent risk factors and postoperative liver dysfunction was established.Results Among the 62 patients,ICG R15 was less than 10.0％ in 26 patients,between 10.0％ and 19.0％ in 17 patients,between 20.0％ and 29.0％ in 9 patients,between 30.0％ and 39.0％ in 5 patients,and over 40.0％ in 5 patients.There were 29 patients with a Child-Pugh A grading and 33 patients with a Child-Pugh B grading in the preoperative evaluation of liver function.The Wilcoxon W rank sum test was used to compare the preoperative ICG R15 in patients with Child-Pugh grading A and B separately.The ICG R15 in Child-Pugh grading A patients was significantly lower than those in Child-Pugh B grading patients (P ＜0.05).There were no significant differences in age,gender,history of previous liver diseases,duration of operation,and intraoperative blood loss (P ＞ 0.05) between the normal liver function group and the liver dysfunction group.However,there was a significant difference in the preoperative ICG R15 and preoperative bilirubin levels (P ＜ 0.05) between the two groups.The preoperative ICG R15 and preoperative bilirubin levels were significant risk factors of postoperative hepatic insufficiency.Regression analysis suggested that preoperative ICG R15 level was an independent risk factor of postoperative hepatic insufficiency (P ＜ 0.05).A regression equation:logit(P) =0.185 × preoperative ICG R15-3.152 could be constructed.Conclusions ICG R15 is an ideal clinical indicator for evaluation of preoperative liver functional reserve in patients with HCCA.It predicted the recovery of postoperative liver function.