Hypoxia and aberrant activation of epidermal growth factor receptor (EGFR) are considered important features of various malignancies. However, whether hypoxia can directly trigger EGFR activation and its clinical implications remain unclear. In this study, we demonstrated that in oral cancer, a typical hypoxic tumor, hypoxia can induce chronic but constitutive phosphorylation of wild-type EGFR in the absence of ligands. Oral cancer cell lines exhibit different EGFR phosphorylation responses to hypoxia. In hypoxic HN4 and HN6 cells, ubiquitination-mediated endocytosis, lysosomal sorting, and degradation lead to low levels of EGFR phosphorylation. However, in CAL-27 and HN30 cells, a novel HIF-1α-induced long noncoding RNA (lncRNA), EUDAL, can compete with the E3 ligase/adaptor complex c-Cbl/Grb2 for binding to EGFR, stabilizing phosphorylated EGFR (pEGFR) and resulting in sustained activation of EGFR and its downstream STAT3/BNIP3 signaling. STAT3/BNIP3-mediated autophagy leads to antitumor drug resistance. A high EUDAL/EGFR/STAT3/autophagy pathway activation predicts poor response to chemotherapy in oral cancer patients. Collectively, hypoxia can induce noncanonical ligand-independent EGFR phosphorylation. High EUDAL expression facilitates sustained EGFR phosphorylation in hypoxic tumor cells and leads to autophagy-related drug resistance.
Humans ; ErbB Receptors/metabolism* ; Mouth Neoplasms/pathology* ; RNA, Long Noncoding/genetics* ; Drug Resistance, Neoplasm/genetics* ; Cell Line, Tumor ; Phosphorylation ; Signal Transduction ; STAT3 Transcription Factor/metabolism* ; Cell Hypoxia ; Autophagy ; Proto-Oncogene Proteins c-cbl/metabolism*

Objective:To evaluate the predictive efficacy of sinus CT radiomics for treatment outcomes in nasal polyp patients undergoing endoscopic sinus surgery.Methods:A retrospective cohort study was conducted at the First Affiliated Hospital of Sun Yat-sen University, including 194 patients with nasal polyps treated between January 2015 and December 2019. The cohort comprised 132 males and 62 females, aged 16 to 75 years. Patients were divided into a training set ( n=135) and an internal validation set ( n=59). An external validation set ( n=34), consisting of 22 males and 12 females aged 16 to 59 years, was included from January 2020 to December 2021. Disease control was evaluated using the criteria from the European Position Paper on Rhinosinusitis and Nasal Polyps 2020 (EPOS 2020). Radiomic features were extracted from sinus CT images and analyzed using the least absolute shrinkage and selection operator (LASSO) regression. Models combining radiomic and clinical features were developed to predict treatment efficacy. Results:The radiomics and combined models, based on four selected features, outperformed the clinical feature model in the training set, with AUC values of 0.901 and 0.915, versus 0.874, respectively. In the internal validation set, AUCs were 0.839, 0.832, and 0.716. Despite reduced AUCs in the external set, the radiomics model maintained good generalizability (0.748, 0.764, 0.620). Decision curve analysis showed significant clinical benefits in both radiomics and combined models.Conclusion:The CT-based radiomics model demonstrates significant predictive power in identifying refractory nasal polyps, suggesting its potential for clinical application in treatment outcome prediction.

Objective:To explore the clinical phenotype and genetic characteristics of a child with Hypotrichosis 14.Methods:A child who had presented at the Henan Provincial People′s Hospital on May 4, 2020 due to hair thinning was selected as the study subject. Clinical data of the child was collected. Peripheral venous blood samples were collected from the child and her parents. Genomic DNA was extracted and subjected to whole exome sequencing. Candidate variants were validated by Sanger sequencing and bioinformatic analysis.Results:The child, a 5-year-old female, had presented with thin, soft lanugo-like hair which was easy to fall off. The child was found to harbor compound heterozygous missense variants of the LSS gene, namely c. 1609G>A (p.V537M) in exon 17 and c. 802T>G (p.F268V) in exon 8, which were respectively inherited from her father and mother. Both variant sites were highly conserved, though based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), both variants were rated as variants of unknown significance (PM2_Supporting+ PP3+ PP4). Conclusion:The c. 1609G>A (p.V537M) and c. 802T>G (p.F268V) compound heterozygous variants of the LSS gene probably underlay the clinical phenotype in this patient.

Objective To explore the value of multi-parameter spectral CT for differentiating grade G2-3 pancreatic neuroendocrine tumor(pNET)and pancreatic neuroendocrine carcinoma(pNEC).Methods Preoperative double-layer detector spectral CT(DLCT)data of 35 patients with pNET(pNET group,including 25 cases of G2 grade and 10 cases of G3 grade)and 17 patients with pNEC(pNEC group)were retrospectively analyzed.Conventional CT and spectral CT parameters were compared between groups,and those being significant different between groups according to univariate analysis were respectively incorporated into multivariate logistic regression to select the independent predictors for identifying grade G2-3 pNET and pNEC.Conventional CT model and spectral CT model were constructed,and the combined model was constructed based on the two.The efficacy of each model for distinguishing grade G2-3 pNET and pNEC was evaluated.Results CT values of lesions during venous phase(OR=0.939,P=0.025)and vascular invasion(OR=5.049,P=0.027)shown on conventional CT were both independent predictors,and conventional CT model was constructed,its area under the curve(AUC)for distinguishing grade G2-3 pNET and pNEC was 0.808.Normalized iodine concentration during venous phase(OR=0.603)and normalized effective atomic number during venous phase(OR=0.847)on spectral CT were both independent predictors(both P＜0.05),and spectral CT model was constructed.The AUC of spectral CT model was 0.894,higher than that of conventional CT model(Z=2.127,P=0.033).The AUC of combined model was 0.924,higher than that of conventional CT model(Z=2.302,P=0.021)but not significantly different with that of spectral CT model(Z=0.827,P=0.408).Conclusion Multi-parameter spectral CT could effectively differentiate grade G2-G3 grade pNET and pNEC.

Objective:To evaluate the effect of sodium sivelestat on the expression of specialized pro-resolving mediators (SPMs) synthesis enzymes in mice with lipopolysaccharide (LPS)-induced acute lung injury (ALI).Methods:Eighteen SPF healthy male C57BL/6 mice, aged 6-8 weeks, weighing 20-25 g, were divided into 3 groups ( n=6 each) using a random number table method: control group (C group), LPS-induced ALI group (ALI group), and LPS-induced ALI + sodium sivelestat group (ALI+ SV group). ALI was induced by intravenous injection of LPS 15 mg/kg through the tail vein. Sodium sivelestat 50 mg/kg was intraperitoneally injected at 1 h after LPS administration. At 12 h after LPS administration, blood samples were collected from the eyeballs for routine blood tests, and the remaining blood was processed for serum extraction. The mice were sacrificed after anesthesia, and lung tissues were collected to determine the wet/dry weight (W/D) ratio, serum concentrations of interleukin-1beta (IL-1β) and IL-10 (by enzyme-linked immunosorbent assay), expression of neutrophil elastase (NE) and SPMs synthesis enzymes 5-lipoxygenase (5-LOX), 12-lipoxygenase (12-LOX), and 15-lipoxygenase (15-LOX) in lung tissues (by Western blot) and to examine the pathological changes of lung tissues which were scored. Results:Compared with C group, the lung injury scores, W/D ratio, white blood cell counts, percentage of neutrophil, and serum IL-1β and IL-10 concentrations were significantly increased, the expression of NE was up-regulated, and the expression of 5-LOX, 12-LOX and 15-LOX was down-regulated in ALI group ( P<0.05). Compared with ALI group, the lung injury scores, W/D ratio, white blood cell counts, percentage of neutrophil, and serum IL-1β concentration were significantly decreased, the serum IL-10 concentration was increased, the expression of NE was down-regulated, and the expression of 5-LOX, 12-LOX and 15-LOX was up-regulated in ALI+ SV group ( P<0.05). Conclusions:The mechanism by which sodium sivelestat alleviates LPS-induced ALI may be related to up-regulating the expression of SPMs synthesis enzyme and promoting the resolution of pulmonary inflammation in mice.

Objective:To investigate the effect of nuclear factor E2-related factor 2 (Nrf2) on the cellular tight junction protein Claudin-18 in endotoxin-induced acute lung injury (ALI).Methods:Eighteen healthy male C57BL/6 mice were divided into control group, endotoxin-induced ALI model group (ALI group) and Nrf2 activator tert-butylhydroquinone (tBHQ) pretreatment group (tBHQ+ALI group) according to random number table method, with 6 mice in each group. Mice endotoxin-induced ALI model was reproduced by intraperitoneal injection of lipopolysaccharide (LPS, 15 mg/kg), and the mice in the control group was injected with an equal amount of phosphate buffer solution (PBS). The mice in the tBHQ+ALI group received three intraperitoneal injections of tBHQ (a total of 50 mg/kg) at an interval of 1 hour before molding. The last injection of tBHQ was accompanied by LPS of 15 mg/kg. The mice in the control group and model group were given equal amounts of PBS, and PBS or LPS was given at the last injection. The mice were sacrificed at 12 hours after LPS injection to take lung tissues. After the lung tissue was stained with hematoxylin-eosin (HE) staining, the pathological changes were observed under light microscopy, and the lung injury score was calculated. The lung wet/dry ratio (W/D) was determined. Nrf2 protein expression in the lung tissue was detected by Western blotting. Positive expression of Claudin-18 in the lung tissue was determined by immunohistochemistry.Results:The lung tissue showed normal structure, without significant pathological change in the control group. Compared with the control group, the alveolar septum widened accompanied by inflammatory cell infiltration, capillary hyperemia and tissue edema in the ALI group, the lung injury score and lung W/D ratio were significantly increased (lung injury score: 6.50±1.05 vs. 1.83±0.75, lung W/D ratio: 3.79±0.22 vs. 3.20±0.14, both P < 0.01), and the Nrf2 protein expression and Claudin-18 positive expression in the lung tissue were significantly lowered [Nrf2 protein (Nrf2/β-actin): 0.41±0.33 vs. 1.22±0.33, Claudin-18 ( A value): 0.28±0.07 vs. 0.44±0.10, both P < 0.05]. After tBHQ pretreatment, the degree of lung histopathological injury was significantly reduced compared with the ALI group, the alveolar space slightly abnormal, inflammatory cell infiltration and tissue edema reduced, the lung injury score and lung W/D ratio were significantly decreased (lung injury score: 3.00±0.89 vs. 6.50±1.05, lung W/D ratio: 3.28±0.19 vs. 3.79±0.22, both P < 0.01), and Nrf2 protein expression and Claudin-18 positive expression in the lung tissue were significantly increased [Nrf2 protein (Nrf2/β-actin): 1.26±0.09 vs. 0.41±0.33, Claudin-18 ( A valure): 0.45±0.04 vs. 0.28±0.07, both P < 0.05]. Conclusion:Nrf2 alleviated pulmonary edema and improved endotoxin-induced ALI by up-regulating Claudin-18 expression.

Objective:To investigate the current situation of the use of transjugular intrahepatic portosystemic shunt (TIPS) for portal hypertension, which should aid the development of TIPS in China.Methods:The China Portal Hypertension Alliance (CHESS) initiated this study that comprehensively investigated the basic situation of TIPS for portal hypertension in China through network research. The survey included the following: the number of surgical cases, main indications, the development of Early-TIPS, TIPS for portal vein cavernous transformation, collateral circulation embolization, intraoperative portal pressure gradient measurement, commonly used stent types, conventional anticoagulation and time, postoperative follow-up, obstacles, and the application of domestic instruments.Results:According to the survey, a total of 13 527 TIPS operations were carried out in 545 hospitals participating in the survey in 2021, and 94.1% of the hospital had the habit of routine follow-up after TIPS. Most hospitals believed that the main indications of TIPS were the control of acute bleeding (42.6%) and the prevention of rebleeding (40.7%). 48.1% of the teams carried out early or priority TIPS, 53.0% of the teams carried out TIPS for the cavernous transformation of the portal vein, and 81.0% chose routine embolization of collateral circulation during operation. Most of them used coils and biological glue as embolic materials, and 78.5% of the team routinely performed intraoperative portal pressure gradient measurements. In selecting TIPS stents, 57.1% of the hospitals woulel choose Viator-specific stents, 57.2% woulel choose conventional anticoagulation after TIPS, and the duration of anticoagulation was between 3-6 months (55.4%). The limitation of TIPS surgery was mainly due to cost (72.3%) and insufficient understanding of doctors in related departments (77.4%). Most teams accepted the domestic instruments used in TIPS (92.7%).Conclusions:This survey shows that TIPS treatment is an essential part of treating portal hypertension in China. The total number of TIPS cases is far from that of patients with portal hypertension. In the future, it is still necessary to popularize TIPS technology and further standardize surgical indications, routine operations, and instrument application.

Background/Aims: To evaluate the causal correlation between complement components and non-viral liver diseases and their potential use as druggable targets. Methods: We conducted Mendelian randomization (MR) to assess the causal role of circulating complements in the risk of non-viral liver diseases. A complement-centric protein interaction network was constructed to explore biological functions and identify potential therapeutic options. Results: In the MR analysis, genetically predicted levels of complement C1q C chain (C1QC) were positively associated with the risk of autoimmune hepatitis (odds ratio 1.125, 95% confidence interval 1.018–1.244), while complement factor H-related protein 5 (CFHR5) was positively associated with the risk of primary sclerosing cholangitis (PSC;1.193, 1.048– 1.357). On the other hand, CFHR1 (0.621, 0.497–0.776) and CFHR2 (0.824, 0.703–0.965) were inversely associated with the risk of alcohol-related cirrhosis. There were also significant inverse associations between C8 gamma chain (C8G) and PSC (0.832, 0.707–0.979), as well as the risk of metabolic dysfunction-associated steatotic liver disease (1.167, 1.036–1.314). Additionally, C1S (0.111, 0.018–0.672), C7 (1.631, 1.190–2.236), and CFHR2 (1.279, 1.059–1.546) were significantly associated with the risk of hepatocellular carcinoma. Proteins from the complement regulatory networks and various liver diseaserelated proteins share common biological processes. Furthermore, potential therapeutic drugs for various liver diseases were identified through drug repurposing based on the complement regulatory network. Conclusions Our study suggests that certain complement components, including C1S, C1QC, CFHR1, CFHR2, CFHR5, C7, and C8G, might play a role in non-viral liver diseases and could be potential targets for drug development.

Objective:To evaluate the role of heme oxygenase-1 (HO-1) in endotoxin-induced acute lung injury (ALI) and the relationship with the regulation of mitochondrial quality control in mice.Methods:Clean-grade healthy male adult C57BL/6 mice, aged 6-8 weeks, weighing 20-25 g, were selected.HO-1 inducible gene knockout mice (HO-1 -/-) were prepared based on CRISPER/Cas9-mediated EGE system, and HO-1 gene overexpression mice (HO-1 + /+ ) were prepared by transfection of HO-1 overexpressed adenovirus vector.The mice were divided into 2 groups ( n=6 each) using a random number table method: control group (group WT, group HO-1 -/-, group HO-1 + /+ ) and endotoxin-induced ALI group (group ALI, group HO-1 -/-+ ALI, group HO-1 + /+ + ALI). Lipopolysaccharide 15 mg/kg was injected through the tail vein to develop the model of endotoxin-induced ALI, and the equal volume of normal saline was given instead in each control group.The mice were sacrificed by bloodletting at 12 h after lipopolysaccharide or normal saline administration.The lung tissues were harvested for microscopic examination of the pathological changes which were scored, for determination of GSH and GSSG contents, for observation of the ultrastructure of mitochondria (with a transmission electron microscope) and survival within 12 h, for measurement of mitochondrial membrane potential (MMP) levels, and for determination of the expression of mitochondrial quality control-related proteins mitochondrial fusion protein 2 (Mfn2) and dynamin-related protein 1 (Drp1), peroxisome proliferator-activated receptor gamma coactivator 1α (PGC-1α), nuclear respiratory factor 1 (NRF1), mitophagy marker protein PTEN-induced kinase 1 (PINK1) and E3 ubiquitin-protein ligase Parkin.The ratio of GSH/GSSG was calculated. Results:Compared with control group (group WT, group HO-1 + /+ and group HO-1 -/-), the 12-h survival rate and MMP were significantly decreased, the lung injury score was increased, GSH content and GSH/GSSG ratio were decreased, and the content of GSSG was increased in endotoxin-induced ALI groups (group ALI, group HO-1 + /+ + ALI and group HO-1 -/-+ ALI) ( P<0.05). Compared with group ALI, the 12-h survival rate and MMP were significantly decreased, the lung injury score was increased, the GSH content and GSH/GSSG ratio were decreased, the GSSG content was increased, and the expression of HO-1, Mfn2, PGC-1α, NRF1, PINK1 and Parkin was down-regulated, and Drp1 expression was up-regulated in group HO-1 -/-+ ALI, and 12-h survival rate and MMP were significantly increased, lung injury score was decreased, GSH content and GSH/GSSG ratio were increased, GSSG content was decreased, the expression of HO-1, Mfn2, PGC-1α, NRF1, PINK1 and Parkin was up-regulated, and the expression of Drp1 was down-regulated in group HO-1 + /+ + ALI ( P<0.05). Conclusions:HO-1 is involved in the process of endotoxin-induced ALI in mice, which is related to the regulation of mitochondrial quality control.

Objective:To evaluate the effect of transcutaneous electrical acupoint stimulation (TEAS) on mitochondrial quality control during endotoxin-induced acute lung injury (ALI) in mice.Methods:Twenty-four clean-grade healthy male C57BL/6J mice, aged 4-6 weeks, weighing 15-20 g, were divided into 4 groups ( n=6 each) according to the random number table method: control group (group C), endotoxin-induced ALI group (group L-ALI), endotoxin-induced ALI plus acupoint electroacupuncture group (group L-ALI+ EA), and endotoxin-induced ALI plus non-acupoint electroacupuncture group (group L-ALI+ SEA). Lipopolysaccharide (LPS) 15 mg/kg was injected via the caudal vein to develop the model of endotoxin-induced ALI in anesthetized mice.In group L-ALI+ EA, at 5 days before LPS injection, bilateral Zusanli and Feishu acupoints were stimulated with an electric stimulator for 30 min each time at a voltage of 1-2 mA and a frequency of 2/15 Hz until the end of the experiment.In group L-ALI+ SEA, stimulation was performed at the points 0.5 cm lateral to the acupoints of bilateral Zusanli and Feishu non-meridian and non-acupoint sites using the shallow puncture method, and the other treatment methods were the same as those previously described in group EA.Group C received no treatment.The mice were sacrificed by euthanasia at 12 h after LPS administration, and lung tissues were obtained for microscopic examination of the pathological changes (with a light microscope) and structure and morphology of mitochondria (with a transmission electron microscope) and for determination of the levels of reactive oxygen species (ROS) and mitochondrial DNA (mtDNA) and contents of glutathione (GSH) and glutathione oxidized (GSSG). The GSH/GSSG ratio was calculated.The expression of mitochondrial fusion proteins mitofusin 1 (Mfn1), Mfn2, optic atrophy1 (OPA1), dynamin-related protein 1 (Drp1), fission protein 1 (Fis1), peroxisome-proliferator-activated receptor γ coactivator-1α (PGC-1α), nuclear respiratory factor-1 (NRF1), NRF2, PTEN-induced putative protein kinase 1 (PINK1) and the E3 ubiquitin ligase (Parkin) was determined by Western blot. Results:Compared with group C, the level of ROS and contents of GSSG and mtDNA were significantly increased, GSH content and GSH/GSSG ratio were decreased, the expression of Mfn1, Mfn2, OPA1, NRF1, NRF2 and PGC-1α was down-regulated, and the expression of Drp1, Fis1, PINK1 and Parkin was up-regulated in L-ALI, L-ALI+ EA and L-ALI+ SEA groups ( P<0.05). Compared with group L-ALI, the level of ROS and contents of GSSG and mtDNA were significantly decreased, GSH content and GSH/GSSG ratio were increased, the expression of Mfn1, Mfn2, OPA1, NRF1, NRF2 and PGC-1α was up-regulated, and the expression of Drp1, Fis1, PINK1 and Parkin was down-regulated in group L-ALI+ EA ( P<0.05). Compared with group L-ALI+ EA, the level of ROS and contents of GSSG and mtDNA were significantly increased, GSH content and GSH/GSSG ratio were decreased, the expression of Mfn1, Mfn2, OPA1, NRF1, NRF2 and PGC-1α was down-regulated, and the expression of Drp1, Fis1, PINK1 and Parkin was up-regulated in group L-ALI+ SEA ( P<0.05). Conclusions:TEAS can reduce endotoxin-induced ALI probably through regulating mitochondrial quality control in mice.