OBJECTIVE:To explore the role of multi-sequence magnetic resonance imaging (MRI) images in preoperative prediction of lymph node metastasis in laryngeal squamous cell carcinoma (LSCC). METHODS:Patients with LSCC undergoing open surgery and lymph node dissection were enrolled (n = 224 training, n = 96 testing). Radiomic features (n = 2394) were extracted from T1-enhanced and T2-weighted images. Features were screened using least absolute shrinkage and selection operator (LASSO) regression, and the best-performing classification model was identified among Logistic Regression, Random Forest, Extreme Gradient Boosting, and Light Gradient Boosting Machine. An imaging biomarker-based nomogram integrating radiomic and clinical features was developed via logistic regression. RESULTS:LASSO regression identified 14 stable features (6 from T1-enhanced images, 8 from T2-weighted). The Random Forest model showed the best radiomics-only performance (area under the receiver operating characteristic curve [AUC]: 0.877 training; 0.875 testing). The combined clinical - radiomics nomogram achieved higher discrimination (AUC: 0.942 training; 0.908 testing), outperforming standalone clinical or radiomic models. CONCLUSION:The radiomic-clinical nomogram enhances preoperative prediction of cervical lymph node metastasis in LSCC, offering the potential to optimize clinical decision-making.

Marine-derived fungi are prolific producers of structurally diverse secondary metabolites with significant pharmaceutical potential. The discovery of natural products has been unprecedentedly accelerated by the prediction of biosynthetic gene clusters (BGCs) within the whole-genome context. This study presents the whole-genome sequencing, comprehensive annotation, and biosynthetic potential predictions of the sponge-associated fungus Aspergillus puulaauensis Hmp-F48. Genome sequencing of A. puulaauensis Hmp-F48 generated a high-quality draft assembly of 35.86 Mb. Structural annotation revealed a complex genomic architecture, comprising 10,611 protein-coding genes, 210 non-coding RNAs, and 155 tRNAs. Functional annotation using NR, Swiss-Prot, GO, KEGG, and eggNOG databases highlighted significant enrichment in biosynthetic, metabolic, and transport processes. AntiSMASH analysis identified 78 putative BGCs, including 16 type I polyketide synthase (T1PKS), 27 nonribosomal peptide synthetase (NRPS), 7 hybrid PKS-NRPS, 9 terpene-related clusters, 6 RiPP-related clusters and 13 clusters associated with other secondary metabolites. Several clusters exhibited high homology to known BGCs responsible for bioactive secondary metabolites, including asperthecin, sterigmatocystin, calbistrins, F-9775 A/B, nidulanin A, aspercryptins, fellutamide B, acetylaranotin, burnettramic acid A, equisetin, and pyranonigrin E. Experimental isolation confirmed the presence of PKS-derived metabolites, including sterigmatocystin, averantin, and decumbenones — one of which represents a previously undescribed congener. This study highlights the extensive biosynthetic potential of A. puulaauensis Hmp-F48, offering valuable insights into its capacity for secondary metabolite production.

In the treatment of malignant tumors, dual-target inhibitors can effectively avoid the compensatory activation of other signaling pathways caused by single-target administration, as well as the pharmacokinetic changes and poor patient compliance associated with co-administration. For this study, we selected phosphatidylinositol 3-kinase (PI3K) and histone deacetylase (HDAC), two critical targets in tumor cell development, and designed dual inhibitors displaying various types and lengths of linkage chains. A quinoline-linked sulfonamide-pyridine fragment as the pharmacophore for PI3K inhibition, and an o-aminobenzamide fragment as the pharmacophore for the Zn2+-binding group (ZBG) of HDAC have been chosen in order to maximize the pharmacokinetic properties of both targets. Using these linkage chains, we designed 41 novel quinoline-linked sulfonamide-pyridine PI3K/HDAC dual-target inhibitors with unique structures. Most of these compounds exhibited strong antiproliferative effects on Jurkat, K562, MCF-7, and PC9R cells. Specifically, SJY26 not only demonstrated potent antiproliferative activity against tumor cells but also exhibited outstanding inhibitory activity against PI3Kα and HDAC1. Moreover, SJY26 significantly inhibited the migration of PC9R cells at 1.25 μM, reduced AKT phosphorylation, and decreased histone H3 deacetylation. In summary, this research highlights the promising therapeutic potential of novel quinoline-linked sulfonamide-pyridine derivatives as PI3K/HDAC dual-target inhibitors, warranting further investigation.

BACKGROUND:Tracheoesophageal fistula (TEF) is a life-threatening complication of advanced esophageal squamous cell carcinoma (ESCC). Cervical ESCC is rare and frequently diagnosed at an advanced stage. Managing cervical esophageal cancer (CEC) is challenging, requiring intervention by a multidisciplinary team (MDT) and innovative surgical management. CASE SUMMARY:Here, we present a 59-year-old male patient with a 5-month history of CEC and difficulty eating for over 20 days, who developed TEF secondary to recurrent ESCC after chemoradiotherapy. He underwent total pharyngolaryngoesophagectomy, left thyroidectomy, and lymphadenectomy. Gastric pull-up was performed to restore gastrointestinal continuity, and a 7 cm × 5 cm supraclavicular artery island flap (SCAIF) was used to reconstruct the lower tracheal defect. Despite severe postoperative complications, he recovered by successful management by a MDT. A 7 cm × 6 cm pectoralis major myocutaneous flap was successfully used to repair the necrotic gastric conduit defect. The patient recovered, regaining the ability to eat and breathe effectively. At the 27-month follow-up, he was alive without recurrence or metastasis. CONCLUSION:This study highlights the efficacy of gastric pull-up and SCAIF reconstruction in managing TEF secondary to recurrent ESCC.

BackgroundGastric cancer (GC) is one of the most common malignant tumors with poor survival. Although cisplatin is a first-line chemotherapy drug for GC, it still has the potential to develop drug resistance and side effects. Miltirone, extracted from Chinese herb Salvia miltiorrhiza Bunge, has been reported to significantly inhibit some types of cancer. However, its effects on GC have not been studied, the possible anti-tumor effects of miltirone in combination with cisplatin in GC patients have not been explored.Materials and methodsHuman GC cell lines AGS, HGC27, MKN45 and MGC803 cells were treated with miltirone and cisplatin individually or combinatorially. Cell proliferation assay, flow cytometric assay, colony formation assay and Western blot were employed to evaluate the cytotoxic effects under these treatments. Wound healing and transwell assays were used to examine the effects of miltirone and/or cisplatin on GC cell migration and invasion. RNA-seq analysis was used to determine miltirone’s potential target genes in AGS cells. GO analysis and molecular docking assay were used to determine the pathways affected by miltirone. Next, we examined changes in the selected pathway proteins. The in vivo animal model was verified the results of the in vitro experiments.ResultsMiltirone inhibited cell growth, migration, and invasion, as well as induced apoptosis in GC cells. In combinatorial treatments, miltirone synergistically enhanced cytotoxicity of cisplatin in GC cells. Moreover, the expression levels of 606 genes appeared to be significantly modulated by miltirone via RNA-seq analyses, and PI3K/AKT signaling pathway was found to refer to miltirone activity. Furthermore, miltirone together with cisplatin treatment significantly reduced the expression levels of p-PI3K, p-Akt, p-mTOR, while the total levels of PI3K and Akt remained unchanged. In addition, compared with the control group, the tumors growth was significantly suppressed in groups treated with the two agents alone or in combination, and even more so in the combination group in vivo.DiscussionMiltirone inhibited the proliferation of GC cells and significantly potentiates the anticancer activities of cisplatin by downregulating the PI3K/AKT signaling pathway. Combination therapy of miltirone and cisplatin represents a novel potential treatment of gastric cancer.

PURPOSE:Vocal fold leukoplakia (VFL) is the most common precancerous lesion of laryngeal carcinoma, with variable malignant potential. The pathogenesis of VFL remains incompletely understood. Increasing evidence indicates that laryngopharyngeal reflux, especially bile acid reflux, may drive epithelial transformation. METHODS:ATP4A and ATP4B expression were assessed by immunohistochemistry in 58 VFL and 15 laryngeal squamous cell carcinoma (LSCC) specimens. Primary VFL epithelial cells were treated with graded concentrations of deoxycholic acid (DCA, 50-400 μM). Small interfering RNAs targeting ATP4A and TGR5, as well as the JAK2/STAT3 inhibitor AG490, were employed to delineate signaling mechanisms. Proliferation and migration were measured by cell counting kit (CCK)-8 and Transwell assays, respectively, while mRNA and protein levels were analyzed using quantitative real-time-PCR and Western blotting. RESULTS:ATP4A positivity was significantly higher in LSCC than in VFL (47.57 ± 15.75% vs 33.47 ± 21.22%, (P = 0.018), whereas ATP4B showed no significant difference. In vitro, DCA at 0-200 μM dose-dependently increased proliferation, migration, ATP4A expression, and TGR5/JAK2/STAT3 phosphorylation, while higher concentrations (300-400 μM) suppressed cell activity. ATP4A knockdown reversed DCA-induced proliferation and migration. Similarly, TGR5 silencing or AG490 treatment abrogated DCA-mediated upregulation of ATP4A and functional enhancement. CONCLUSIONS:DCA activates the TGR5/JAK2/STAT3 axis to upregulate ATP4A, thereby promoting VFL epithelial cell proliferation and migration. These findings provide novel mechanistic insight linking bile acid reflux to VFL progression and identify TGR5/ATP4A as potential therapeutic targets.

Acute lung injury (ALI), characterized by uncontrolled inflammatory responses, is a critical condition with limited clinical treatment options. Activation of the NLRP3 inflammasome represents a pivotal event in ALI pathogenesis. Although targeting NLRP3 (The NOD-like receptor (NLR) family member containing a pyrin domain 3) inflammasome has been considered to be a potential therapy, the underlying mechanism through which pathway the pulmonary inflammation is modulated remains controversial. Here, we demonstrate that 475A, a highly selective ROCK2 (Rho-associated coiled-coil forming protein kinases 2) inhibitor, effectively attenuates pulmonary inflammation and alleviates lung injury. Mechanistically, 475A promotes NLRP3 protein ubiquitination and subsequent proteasomal degradation by disrupting YAP (Yes-associated protein)-NLRP3 protein interactions, thereby suppressing inflammatory cytokine production in macrophages. This study provides the first evidence linking ROCK2 inhibition to enhanced NLRP3 degradation in ALI treatment, paving the way for developing 475A as a therapeutic agent and establishing the ROCK2/YAP/NLRP3 axis as a novel strategy for treating NLRP3-driven inflammatory diseases.

PURPOSE:The expression of MAGE-A9 in vocal fold leukoplakia (VFL) tissues and the mechanism underlying its role in the malignant transformation of VFL remain unclear. This study investigated the role of MAGE-A9 in the proliferation, migration, and invasion of VFL epithelial cells as well as the underlying regulatory mechanism. METHODS:MAGE-A9 expression in VFL tissues was detected by immunohistochemistry. The CCK-8 assay, flow cytometry, and transwell assays were performed to determine the viability, cell cycle distribution and apoptosis, as well as migration and invasion, respectively, of primary cultured VFL epithelial cells. Ki67, cell cycle proteins, and proteins in the NF-kB-MMP-2/9 pathway were assessed by Western blotting. RESULTS:MAGE-A9 expression was detected in 54.5% (18/33) of vocal fold polyps, 48.9% (44/90) of VFLs, and 84.8% (28/33) of laryngeal cancers. Significantly higher levels of expression were found in laryngeal cancer than in vocal fold polyps and VFL tissues (P < 0.001). The expression of MAGE-A9 tended to increase with the severity of VFL dysplasia. In VFL epithelial cells, the overexpression of MAGE-A9 significantly increased the viability, proliferation, migration, and invasion of the cells and reduced the level of apoptosis. The cell cycle effects of MAGE-A9 overexpression included an increased proportion of cells in the G2 and S phases and a decreased proportion of those in the G1 phase (P = 0.002), leading to an altered G1/S phase transition. MAGE-A9 overexpression also significantly increased p-IKBα, p-p65, MMP2, and MMP9 levels while decreasing those of IKBα. All of the effects of MAGE-A9 were inhibited by treating the cells with the IκBα phosphorylation inhibitor BAY 11-7082. CONCLUSION:MAGE-A9 expression tended to increase with the severity of dysplasia in VFL and was significantly higher in laryngeal cancer than in VFL. MAGE-A9 was shown to promote the proliferation, migration, and invasion of VFL epithelial cells via the NF-kB pathway and downstream targets such as MMP-2/9.

Wound healing remains a significant global health challenge, affecting millions annually and imposing substantial economic burdens. Most commercially available biomaterials for wound management primarily address external symptoms, including hemostasis, exudation, scarring, and infection. Advanced biomaterials derived from endogenous molecules aim to better replicate the native wound microenvironment, promoting enhanced repair. Since wounds frequently occur on exposed skin, which is vulnerable to UVA radiation and requires protective yet invisible materials, traditional wound care products often lack these essential features. Inspired by natural UV protection mechanisms, a novel bioscaffold is developed using recombinant human collagen XVII (rhCOL17) crosslinked with porphyra-334 (P334) to improve wound healing under UVA exposure. The resulting rhCOL17-P334 conjugate integrates extracellular matrix (ECM)-like cues with UVA-shielding properties provided by P334. This conjugate is used to construct a transparent, injectable hydrogel combining gelatin methacryloyl (GelMA) and rhCOL17-P334 (GCP). GCP significantly inhibits UVA-induced fibroblast senescence and improves wound healing outcomes by targeting integrin α6β4 through rhCOL17. Its transparency facilitates convenient wound monitoring while also addressing the aesthetic requirement for invisibility. By combining UVA shielding with wound repair capabilities, GCP presents a promising platform for advanced wound management.

All raw data for the analyses and visualizations in this study.

FMS-like tyrosine kinase 3 (FLT3) is the most frequently mutated gene in acute myeloid leukemia (AML), and is associated with poor prognosis and a high relapse rate. Gilteritinib, a second-generation FLT3 inhibitor, is an important target drug for treating patients with FLT3-internal tandem duplication (ITD) AML, is approved for the treatment of relapsed/refractory FLT3-mutant acute myeloid leukemia, although challenges such as drug resistance and reduced potency remain. Herein, mitoxantrone-liposomes sensitized FLT3-ITD AML cells to gitretinib both in vivo and in vitro. RNA-sequencing revealed that combination treatment resulted in specific changes in gene expression as well as predicted the mechanism. Primary AML cells harvested from patients with FLT3-ITD AML showed a significant response to combination treatment in vitro. Our data suggests a novel and promising therapeutic strategy for patients with FLT3-ITD AML and relapsed/refractory FTL3-ITD AML.

This study aimed to analyse the relationship of the blood lipid profile and interleukin-6 (IL-6) with osteoporosis and osteopenia and to explore the predictive value of the combined application of these biomarkers in osteoporosis and osteopenia. Data from 276 patients treated in the orthopaedics department were retrospectively analysed. Their general information was collected, and the relationships among the blood lipid profile, IL-6 with bone turnover markers, and bone mineral density (BMD) were analysed. Patients were categorized based on their T scores for intergroup comparisons. Finally, the diagnostic efficiency of lipid metabolism markers and IL-6 for osteoporosis and osteopenia was assessed using receiver operating characteristic (ROC) curves. (1) In both males and females, a negative relationship was observed between BMD and several biomarkers, including total cholesterol (TC), apolipoprotein B (ApoB), low-density lipoprotein cholesterol (LDL-C), free fatty acids (FFAs), and IL-6. Additionally, apolipoprotein A1 (ApoA1) was negatively correlated with BMD only in females, and the ApoA1/ApoB ratio was positively correlated with BMD only in males. (2) FFAs and IL-6 were positively correlated with β-CrossLaps peptide in males. However, for females, TC, ApoB, LDL-C, and IL-6 were negatively correlated with 25-hydroxyvitamin D. FFAs, IL-6, and age were negatively correlated with osteocalcin in males and females. (3) According to the T scores for the lumbar spine, the TC, ApoA1, ApoB, HDL-C, LDL-C, FFA, and IL-6 levels in the osteoporosis group and the TC, ApoB, LDL-C, and FFA levels in the osteopenia group were significantly greater than those in the normal bone mass group. Additionally, the osteoporosis group presented substantially higher levels of ApoA1, FFAs, and IL-6 than the osteopenia group. (4) IL-6 was positively correlated with FFAs, while a negative correlation was observed with TC, ApoA1, ApoB, HDL-C, and LDL-C. (5) The ROC curve revealed that the areas under the curve (AUCs) of TC, FFAs, IL-6, ApoA1, and the ApoA1/ApoB ratio for predicting osteoporosis or osteopenia were 0.634, 0.713, 0.670, 0.628, and 0.516, respectively, whereas the AUC of the combination of TC, FFAs, IL-6, and ApoA1 was 0.846, and the AUC of the combination of TC, FFAs, IL-6, and the ApoA1/ApoB ratio was 0.842. In the sex stratification analysis, in males, the AUCs of TC, FFAs, IL-6, and the ApoA1/ApoB ratio for the prediction of osteoporosis or osteopenia were 0.596, 0.688, 0.739, and 0.539, respectively. In contrast, the AUC of the combination of TC, FFAs, IL-6, and the ApoA1/ApoB ratio was 0.838. In females, the AUCs of TC, FFAs, IL-6, ApoA1, and the ApoA1/ApoB ratio for predicting osteoporosis or osteopenia were 0.620, 0.728, 0.653, 0.611, and 0.502, respectively, whereas the AUC of the combination of TC, FFAs, IL-6, and ApoA1 was 0.841, and the AUC of the combination of TC, FFAs, IL-6, and the ApoA1/ApoB ratio was 0.828. The levels of TC, FFAs, IL-6, ApoA1, and ApoB could contribute to changes in bone metabolism, moreover, FFAs could induce an increase in IL-6 further aggravating bone mass loss and leading to osteoporosis. Based on the comparison of the AUC results, the combination of TC, FFAs, and IL-6 with ApoA1 or the ApoA1/ApoB ratio can better predict osteoporosis or osteopenia in patients, and the diagnostic efficiency is significantly better than that of any individual indicator. The regulation of blood lipid levels should become a new target for clinicians to treat osteoporosis and osteopenia.

PURPOSE:The mechanisms driving the progression of vocal cord leukoplakia (VCL) to laryngeal squamous cell carcinoma (LSCC) remain unclear, posing a significant barrier to the effective prevention, early diagnosis, and targeted treatment of LSCC. Therefore, it is essential to characterize the cellular microenvironmental differences between VCL and LSCC at single-cell resolution. EXPERIMENTAL DESIGN:In the study, we conducted single-cell RNA sequencing and spatial transcriptomics on surgical tissue specimens obtained from 36 patients diagnosed with vocal cord polyps (VCP), VCL, and LSCC. RESULTS:Our study generated the first single-cell atlas of VCP and VCL while expanding the cancer cell atlas of LSCC. This dataset comprises 318,907 cells and 12,679 spatial transcriptomic spots, allowing the identification of distinct cellular subclusters. We observed that VCL, as a transitional lesion between benign and malignant states, exhibits a hybrid microenvironment that mirrors VCP and LSCC, with early signs of immunosuppressive activity. Immunoregulatory cell populations demonstrate significant gene expression and functional pathway differences between VCL and LSCC. CONCLUSIONS:Our single-cell RNA sequencing and spatial transcriptomics analyses revealed the cellular heterogeneity underlying benign, precancerous, and malignant laryngeal lesions. We identified epithelial subclusters in VCL with malignant potential and observed shared immunosuppressive features with LSCC, suggesting their role in disease progression. These findings provide valuable insights into the molecular transition from VCL to LSCC and emphasize potential targets for early diagnosis and therapeutic intervention.

The scarcity of ATP-binding cassette subfamily B member 1 (ABCB1, also known as P-glycoprotein, P-gp) inhibitors suitable for clinical application in improving multidrug resistance (MDR) promotes the development of drugs aimed at reversing MDR. In this work, we reported a comprehensive study for the first time about the reversal activity of β-carboline derivatives on ABCB1-mediated MDR. Among 48 synthesized derivatives, compound K27 significantly increased the sensitivity of ABCB1-mediated MDR SW620/AD300 cells to paclitaxel (PTX) (IC50 = 15.33 ± 5.4 nM, RF = 171.2) and hardly showed toxicity even at a high concentration of 20 μM when used alone. The in vitro studies indicated that compound K27 distinctly enhanced the arresting effect of PTX on the SW620/AD300 cell cycle, thereby inhibiting their proliferation. Mechanistically, compound K27 was confirmed to directly bind to ABCB1 to inhibit efflux function, reducing cellular efflux and ensuring stable intracellular concentration of PTX without affecting ABCB1's normal expression. Importantly, the combination of compound K27 and PTX exhibited potent tumor suppression in vivo without generating toxicity. These results demonstrated that β-carboline compounds represented by compound K27 may be potent ABCB1 inhibitors with considerable potential in effectively reversing ABCB1-mediated MDR, showing promising prospects.

Twenty-four cardiac glycosides - including six previously undescribed compounds (1-6) and eighteen known analogues (7-24) - were isolated from the whole herb of Adonis amurensis. Their structures were elucidated through comprehensive spectroscopic analysis (1D/2D NMR, HRMS) and acid hydrolysis. All compounds exhibited potent in vitro cytotoxicity against diverse cancer cell lines, with IC₅₀ values ranging from 0.02 to 2.89 μM. Structure-activity relationship (SAR) studies identified critical structural determinants of potency: C-11 hydroxylation, C-10 substituents, glycosylation at C-3, and oligosaccharide chain length. In vitro, compound 5 induced apoptosis in HCT-116 cells via caspase-3/PARP activation. In vivo, compound 5 (2 mg/kg) significantly (41.5 %) inhibited tumor growth in an HCT-116 xenograft mouse model. Immunofluorescence analysis of tumor tissues revealed decreased Ki67 expression (proliferation) alongside elevated TUNEL, cleaved caspase-3, and BAX levels (apoptosis).

The importance of vocal cord leukoplakia (VCL) in the etiology and progression of laryngeal carcinoma has gained increasing recognition. However, research into the mechanisms of laryngeal precancerous lesions such as VCL, has been hampered by the small size of VCL epithelial cells and their limited culture lifespan. In this study, we enhanced the primary culture protocol for VCL epithelial cells and introduced simian virus 40 Large T to establish an immortalized cell line, designated hVCL-MSDEP01. We confirmed that hVCL-MSDEP01 expresses epithelial-specific genes and proteins; it also demonstrates distinct cell cycle dynamics and apoptosis rates compared with primary cells. In conclusion, hVCL-MSDEP01 serves as an ideal in vitro model for studying VCL. This cell line will substantially advance research into the etiology and progression of laryngeal carcinoma.

Triple-negative breast cancer (TNBC) represents a highly aggressive and heterogeneous malignancy. Currently, multitarget drug approaches present a promising therapeutic approach for TNBC. Utilizing a combinatorial chemistry strategy to construct a virtual screening database, dual ROCK/HDAC-targeting benzothiophene compounds were identified. Notably, compound 10h effectively inhibits ROCK1/2 and HDAC1/2/3/6/8 while demonstrating potent antiproliferative activity against breast cancer cells. In an orthotopic mouse model of breast cancer, 10h significantly suppressed tumor growth without apparent toxicity. Importantly, 10h induced immunogenic cell death (ICD), promoted dendritic cells (DCs) maturation, and activated T cells, thereby initiating antitumor immunity. In conclusion, compound 10h is a novel dual-target ROCK/HDAC inhibitor that represents a promising treatment strategy for TNBC.

OBJECTIVE:We assessed the expression of glucose transporter (GLUT)-1 and hexokinase (HK)-Ⅱ in vocal cord leukoplakia (VCL) and investigated its clinical significance. METHODS:We collected 127 archival pathological specimens from patients with VCL, 29 from vocal cord polyps, and 31 with laryngeal carcinoma at our hospital between December 2018 and December 2022. GLUT-1 and HK-II expression was detected using immunohistochemistry. RESULTS:Significant differences were observed between the VCL and vocal cord polyp groups according to sex (P < 0.0001), smoking status (P = 0.001), and alcohol consumption (P = 0.007). GLUT-1 expression was observed in 36.2%, 20.7%, and 90.3% of patients with VCL, vocal cord polyps, and laryngeal carcinoma, respectively. GLUT-1 was highly expressed in severe dysplasia or carcinoma in situ (CIS) compared with vocal cord polyps (P < 0.001). GLUT-1 expression was lower in the VCL group than in the laryngeal carcinoma group (P < 0.001). HK-II expression was observed in 42.5%, 34.5%, and 80.6% of patients with VCL, vocal cord polyps, and laryngeal carcinoma, respectively. HK-II expression was significantly higher in severe dysplasia or CIS compared with vocal cord polyps (P = 0.004). HK-II expression was lower in the VCL group than in the laryngeal carcinoma group (P < 0.001). Both GLUT-1 expression (P < 0.001) and HK-II expression (P = 0.03) increased significantly with the degree of dysplasia. A significant correlation was observed between GLUT-1 and HK-II expression (r = 0.313, P < 0.001). CONCLUSION:GLUT-1 and HK-II are significantly overexpressed in VCL patients. In VCL patients, GLUT-1 and HK-II expression levels increase proportionally with the degree of dysplasia.