Objective : To construct a molecular classification system for head and neck squamous cell carcinoma (HNSCC) utilizing hypoxia-related gene (HAG) expression profiles, and to comprehensively examine the clinicopathological significance and biological functions of the hypoxia gene stanniocalcin 2 (STC2) in HNSCC. Methods : Transcriptomic data and clinical information of 546 HNSCC samples were obtained from The Cancer Genome Atlas (TCGA) database, and based on the expression profiles of 200 HRGs, HNSCC was classified subclasses using non-negative matrix factorization (NMF). HNSCC was classified into three subclasses (C1, C2, and C3), and the molecular characteristics and prognostic differences of the subclasses were assessed by comparing the tumor mutation load, functional enrichment analysis, drug sensitivity, and clinical features among the subclasses. LASSO-Cox regression was used to screen prognosis-related genes and construct prognostic models. Using oral squamous cell carcinoma (OSCC)-related data in the TCGA database, we analyzed the expression differences of STC2 in OSCC and control samples, and detected the mRNA and protein expression of STC2 in oral squamous carcinoma samples using qRT-PCR and immunohistochemistry. We knocked down STC2 in CAL-27 cells and verified the knockdown efficiency by qRT-PCR and Western blot. CCK-8 assay and cell scratch assay were used to assess the effect of STC2 on cell proliferation and migration ability. Results: Based on HRGs expression profiles, HNSCC was categorized into three subclasses (C1, C2, and C3). Subclass C1 had moderate hypoxic activity and good prognosis; subclass C2 had the highest hypoxic activity, poor prognosis, and poor sensitivity to CTLA-4 inhibitors (P<0.05); subclass C3 had the lowest hypoxic activity and moderate prognosis, and STC2 belonged to subclass C3. The frequency of cyclin-dependent kinase inhibitor 2A (CDKN2A), phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha (PIK3CA), and tumor protein p53 (TP 53) mutations was higher in HNSCC. C1 genomic gain and deletion burden were significantly higher than C3 subclass (P<0.05) and C2 genomic gain than C3 subclass (P<0.05). The C2 subclass was significantly enriched in hypoxia-associated pathways, such as glycine metabolism and base excision repair (P<0.05). The C1, C2, and C3 subclasses were significantly positively correlated in terms of sex (male) (Cramer’s V=0.15), radiation exposure (Cramer’s V=0.12), medication (Cramer’s V=0.18), and pathological grading (G1/G2) (Cramer’s V=0.25) (P<0.05). Nine prognosis-related genes were screened by LASSO-Cox regression, among which high expression of STC2 was positively correlated with poorer overall survival (OS) in HNSCC patients (P<0.01). Bioinformatics analysis showed that STC2 mRNA expression was higher in OSCC than in normal controls (P<0.05). qRT-PCR and immunohistochemistry confirmed that both mRNA and protein expression of STC2 were significantly upregulated in OSCC tissues and cells (P<0.01). In vitro experiments showed that STC2 expression was knocked down to approximately 80% in CAL-27 cells (P<0.001), and the STC2 knockdown group had a reduced value-added rate (P<0.001) and a reduced percentage of scratch closure (P<0.05) compared with the control group. Conclusion We successfully constructed a molecular typing system for HNSCC based on the expression profiles of HRGs and categorized HNSCC into three subclasses with significant prognostic differences, among which the C2 subclass had the highest hypoxic activity and the poorest prognosis. STC2 was highly expressed in HNSCC and suggested a poor prognosis, demonstrating that it may be a potential target for HNSCC treatment.

Neutrophil extracellular traps(NETs)are fibrous web-like structures composed of decondensed chromatin and granular proteins released by neutrophils,with the ability to capture and kill bacteria.Pathogens,such as bacteria and viruses,can trigger the formation of NETs via NETosis,a type of programmed cell death that has two distinct forms:suicidal NETosis and vital NETosis.Numerous studies have found that NETs interact with immune cells in the tumor microenvironment,where they activate macrophages,promote immunosuppressive effects of myeloid-derived suppressor cells,and coat the tumor surface to prevent cytotoxic effects of CD8+T cells and natural killer cells.Recent research has identified a substantial presence of NETs in oral squamous cell carcinoma(OSCC)tissues,indicating a complex re-lationship between NETs and OSCC development.Depending on the phenotype of neutrophils,NETs may exhibit pro-tu-mor or anti-tumor effects.For instance,NETs derived from N1-type neutrophils may exert anti-tumor effects,while TGF-β-induced NETs derived from N2-type neutrophils may exert pro-carcinogenic activity,thereby contributing to the devel-opment of oral squamous metaplasia.Furthermore,NETs likely play a role in OSCC metastasis by capturing circulating tumor cells and inducing a hypercoagulable state,thereby facilitating tumor-related thrombus formation and hematoge-nous metastasis.The involvement of NETs in the occurrence and progression of OSCC opens new avenues for anti-tumor therapy and prognostication.Inhibiting NET formation can significantly suppress the development of chemotherapy-in-duced drug resistance and reduce the risk of thrombosis in OSCC patients,thereby inhibiting tumor metastasis.Current-ly,multiple prognostic models based on NET-related genes have been constructed and validated for head and neck squa-mous cell carcinoma,indicating the potential clinical value of NETs.However,the association between NETs and OSCC treatment is still unclear,necessitating further research on its underlying mechanisms and feasibility.This article attempts to review the relationship between NETs and OSCC,aiming to provide novel perspectives for OSCC treatment.

Objective To study the appropriate concentration of oral Gadovist solution for improving the image quality of three-dimensional sampling perfection with application optimized contrasts using different flip angle evolutions magnetic resonance cholangiopancreatography(3D-SPACE-MRCP).Methods In vitro experiments,0.05％,0.1％,0.15％and 0.2％Gadovist solution and direct drinking water were put into 100 mL plastic containers respectively for 3D-SPACE-MRCP scanning to measure the signal intensity(SI)of Gadovist solutions in each group.The concentration and the SI of Gadovist solution were analyzed with Spearman correlation,and one-way variance analysis was performed to compare the SI of different concentration Gadovist solutions.In clinical experiments,128 subjects were randomly divided into 4 groups,and then taken orally 300 mL of 0.05％,0.1％,0.15％and 0.2％Gadovist solutions respectively.The 3D-SPACE-MRCP scanning was performed,and the image quality was evaluated by two physicians and compared by one-way variance analysis.Results In vitro experiments,there was an extremely strong negative correlation between the concentration and the SI of Gadovist solution(r=-0.969,P＜0.05),and there were significant differences in the SI among different concentrations of Gadovist solution(P＜0.05).In clinical experiments,among the 26 cases with first-level images,24 cases were distributed in the 0.1％Gadovist solution group.There was a statistically significant difference in image quality of the 3D-SPACE-MRCP with different oral concentrations of Gadovist solution(F=89.57,P＜0.05),however,there was no statistically significant difference between the 0.15％and the 0.2％Gadovist solution groups using the Tukey test(P＞0.05).Conclusion Oral Gadovist solution can significantly improve the images quality of 3D-SPACE-MRCP,and 0.1％solution is the appropriate dilution concentration.

Objective To investigate the effect of magnetic resonance cholangiopancreatography(MRCP)after Gd-BOPTA enhancement to clearly show the pancreaticobiliary duct,and to explore the reasonable time of MRCP examination after enhance-ment.Methods The quality of MRCP images in 124 patients after Gd-BOPTA enhancement was retrospectively evaluated by two physicians.The interval time of MRCP examination from the beginning of arterial phase to MRCP acquisition was obtained from pic-ture archiving and communication system(PACS),and the relationship between the image quality grade of pancreaticobiliary duct and the interval time was analyzed by ANOVA.Then the reasonable examination time of MRCP was calculated.Results After Gd-BOPTA enhancement,there was a significant difference in the interval time between different grades of intrahepatic and extrahepatic bile ducts on MRCP images(P＜0.05).The image quality of bile duct decreased with the increasing interval time,however,there was no sig-nificant difference in the interval time between different grades of pancreatic duct(P＞0.05).The mean interval time of clear grade of intra-hepatic bile duct was(9.9±3.1)min,and the 95％confidence interval(CI)was 9.2-10.6 min.Conclusion The clarity of the bile ducts on MRCP is gradually decreased after Gd-BOPTA enhancement,while no change in that of the pancreatic duct.The MRCP examination should be completed within 10.6 min after the arterial phase beginning,which is of great significance for controlling image quality.

Objective:By activating gamma-aminobutyric acid(GABA)neurons in zona incerta(ZI),we observed the alleviation of motor ability impairment in mice treated with lipopolysaccharide(LPS),and investigate the function of GABA neurons in the process of motor function impairment in mice with neuroinflammation.Methods:The central nervous system infection model of C57BU6 mice was eatablished by intraperitoneal injection of LPS.The motor function were detected by open field test and running wheel test in mice.The expression of c-Fos in ZI neurons and ionized calci-um binding adaptor molecule 1(Iba-1)in glial cells were detected by immunofluorescence staining.rAAV-DIO-hM3Dq-mCherry was bilaterally injected into ZI of adult male VGAT-Cre mice through the stereotaxic technique.After three weeks with the virus sufficiently expressing in the neurons,the neuroinflammation model was made by intrabitoneal injection of LPS,and GABAergic neurons were activated by intrabitoneal injection of N-oxyclozapine(CNO)30 min before behavioral tests.The motor ability in mice were detected by open field test and running wheel test.Results:The results of open field experiment and wheel experiment showed that the activity of mice in LPS group was significantly decreased(P＜0.05)compared with the control group.The expression of ZI c-Fos positive protein in the brain of mice was significantly down-regulated with Iba-1 positive protein up-regulating in the ZI.The chemogenetic results showed that the activity of LPS mice was relieved with ZI GABAergic neurons being activated(P＜0.05).Conclusion:The activity of ZI GABAergic neurons was significantly decreased and the motor ability was impaired in mice with neuroin-flammation in the brain.Chemogenetic activation of ZI GABAergic neurons significantly improved the motor ability of inflammatory mice.

Objective:To investigate the independent risk factors for postoperative severe pulmonary infection (SPI) in patients with severe traumatic brain injury (sTBI) and evaluate their predictive value.Methods:A retrospective cohort study was conducted to analyze the clinical data of 163 sTBI patients admitted to Tongji Hospital, Tongji Medical College of Huazhong University of Science and Technology from April 2021 and March 2023, including 101 males and 62 females, aged 20-80 years [53.0(46.0, 59.0)years]. The surgical procedures involved decompressive craniectomy, subdural hematoma removal, epidural hematoma removal, and intracranial hematoma removal. The patients were divided into SPI group ( n=62) and non-SPI group ( n=101) according to whether they had SPI postoperatively. The following data of the two groups were collected, including gender, age, preoperative Glasgow coma scale (GCS), elevated blood glucose, abnormal liver function, abnormal renal function, hemoglobin level, anemia, albumin level, hypoproteinemia, white blood cell count, neutrophil count, lymphocyte count, platelet count, neutrophil-to-lymphocyte ratio (NLR), derived neutrophil-to-lymphocyte ratio (dNLR), platelet-to-lymphocyte ratio (PLR), prognostic nutritional index (PNI) and serum lactate dehydrogenase (LDH) level. All the hematological tests were performed on venous blood samples collected preoperatively before anti-inflammatory treatment. Independent risk factors for predicting the postoperative occurrence of SPI in sTBI patients were identified through univariate analysis and multivariable stepwise regression analysis. The predictive value of separate indicator or indicators combined was assessed by calculating the area under the curve (AUC) of the receiver operating characteristic (ROC) curve. Results:Univariate analysis demonstrated that preoperative GCS, albumin level, lymphocyte count, NLR, PNI and serum LDH level in both groups were significantly correlated with the postoperative occurrence of SPI ( P<0.05), while gender, age, elevated blood glucose, abnormal liver function, abnormal renal function, hemoglobin level, anemia, hypoproteinemia, white blood cell count, neutrophil count, platelet count, dNLR and PLR were not correlated with the postoperative occurrence of SPI in sTBI patients ( P>0.05). Multivariable stepwise regression analysis revealed that low lymphocyte count (95% CI -0.337, -0.013, P<0.05), high NLR (95% CI -0.023, -0.005, P<0.01), low PNI (95% CI 0.007, 0.026, P<0.01), and high serum LDH (95% CI -0.002, -0.001, P<0.01) were independent risk factors for SPI in sTBI patients ( P<0.05). ROC curve analysis indicated that low lymphocyte count, high NLR, low PNI and high serum LDH level could predict SPI in sTBI patients postoperatively, with the combination of PNI and serum LDH showing the highest predictive ability (AUC=0.78, 95% CI 0.70, 0.85). Conclusion:Low lymphocyte count, high NLR, low PNI, and high serum LDH level are independent risk factors for postoperative SPI in patients with sTBI, and the combination of PNI and serum LDH possesses a high predictive value for postoperative SPI in sTBI patients.

Objective:To develop a predictive model for intrapartum fever related to chorioamnionitis in parturients undergoing epidural labor analgesia.Methods:This was a retrospective study. The parturients with intrapartum fever (axillary temperature ≥38 ℃) who received epidural labor analgesia from January 2020 to December 2022 in Sichuan Maternal and Child Health Hospital were selected as model group, and parturients with intrapartum fever who received epidural labor analgesia from January to October 2023 in Sichuan Maternal and Child Health Hospital were selected as validation group. The parturients in model group were divided into histological chorioamnionitis stage ≥Ⅱ group (HCA≥Ⅱ group) and histological chorioamnionitis stage <Ⅱ group (HCA<Ⅱ group) according to the results of placental histopathological examination. Logistic regression analysis was used to screen the independent risk factors for intrapartum fever related to chorioamnionitis in parturients, and then a nomogram model was established. The discrimination of the model was verified by the area under the the receiver operating characteristic curve. The consistency of the model was verified by the calibration curve, and the clinical effectiveness of the model was determined by the decision curve. The validation dataset was used to further evaluate the model.Results:A total of 308 parturients were finally included in model group and 99 parturients in validation group. Multivariate logistic regression analysis showed that the gestational age, meconium-stained amniotic fluid, c-reactive protein concentration and maximum body temperature were independent risk factors for intrapartum fever related to chorioamnionitis in parturients undergoing epidural labor analgesia ( P<0.05). Based on this, a nomogram risk prediction model was developed. The area under the curve (95% confidence interval) was 0.844 (0.744-0.944) in model group and 0.812 (0.674-0.950) in validation group. The calibration curve showed that the prediction probability of the model had good consistency with the actual probability of diagnosis. The decision curve showed that the threshold probability of the prediction model in model group and validation group was 10%-98% and 10%-78%, respectively. Conclusions:A nomogram prediction model for intrapartum fever related to chorioamnionitis is successfully established based on the gestational age, c-reactive protein concentration, meconium-stained amniotic fluid and maximum body temperature in parturients undergoing epidural labor analgesia. The model has good predictive performance and clinical value.

Objective To compare the efficacy of anterior-posterior and posterior-anterior screw fixation for posterior malleolar fractures surgery.Methods A retrospective analysis of 376 cases of posterior malleolar fractures treated with lag screws from January 2011 to October 2022 with more than 12 months of follow-up period was conducted.The patients were divided into two subgroups based on the thickness of the fracture fragment,with 167 cases in the small fracture subgroup having a fracture fragment thickness＜17 mm(screw thread length)and 209 cases in the large fracture subgroup having a fracture fragment thickness ≥ 17 mm.Each subgroup was further divided into anterior-posterior and posterior-anterior groups based on the direction of screw fixation in the posterior malleolar fracture surgery.In the small fracture subgroups,there were 74 cases in the anterior-posterior group and 93 cases in the posterior-anterior group.In the large fracture subgroup,there were 88 cases in the anterior-posterior group and 121 cases in the posterior-anterior group.The American Orthopaedic Foot and Ankle Society(AOFAS)ankle-hindfoot score was measured at the last follow-up.The displacement of the fracture fragment in the direction of the fracture line(Dn)and perpendicular to the fracture line(Dt)were measured on the first day after surgery and at the last follow-up,and the displacement of the fracture fragment was calculated,which was the difference between Dn+Dt at the last follow-up and Dn+Dt on the first day after surgery.Results On the first day after surgery,X-ray showed no significant difference in Dn and Dt between the anterior-posterior and posterior-anterior groups in both of the small and large fracture subgroups(P＞0.05).The entire group was followed up for 12-85 months,with an average of 19.3 months.In the small fracture subgroup,the displacement of the fracture fragment in the posterior-anterior group[(0.11±0.19)mm]was superior to that in the anterior-posterior group[(0.19±0.21)mm;P=0.011],and the AOFAS score was also superior to that in the anterior-posterior group[(80.2±8.4)points vs.(76.2±8.6)points,P=0.003].In the large fracture subgroup,there was no significant difference in fracture displacement between the posterior-anterior group[(0.11±0.18)mm]and the anterior-posterior group[(0.12±0.19)mm;P=0.630],and there was also no significant difference in AOFAS scores[(84.1±7.8)points vs.(82.8±7.6)points,P=0.246].Conclusions There is no significant difference in the reduction effect between anterior-posterior and posterior-anterior lag screw internal fixation for posterior malleolar fractures in trimalleolar fractures.For patients with fracture thickness＜17 mm,posterior-anterior fixation is superior to anterior-posterior fixation;for patients with fracture thickness ≥17 mm,there is no significant difference in the efficacy between anterior-posterior and posterior-anterior fixation.

Copper plays an important role in maintaining the normal metabolic function of cells and participates in a variety of physiological processes in cells， including cell respiration， neuropeptide processing， and iron transport， and therefore， it is of great significance to maintain copper homeostasis. In the central nervous system， copper homeostasis is involved in the regulation of synaptic function and the formation of myelin sheath， and copper homeostasis disorder is closely associated with the development of various neurodegenerative diseases. Recent studies have shown that copper homeostasis disorder caused by the mutation of ATPase copper transporting α may lead to the development and progression of amyotrophic lateral sclerosis （ALS）. This article reviews the research advances in copper homeostasis disorder in the pathogenesis of ALS and points out that maintaining copper homeostasis may provide a new target for the treatment of ALS.

FRMD6, a member of the 4.1 ezrin-radixin-moesin domain-containing protein family, has been reported to inhibit tumor progression in multiple cancers. Here, we demonstrate the involvement of FRMD6 in lung cancer progression. We find that FRMD6 is overexpressed in lung cancer tissues relative to in normal lung tissues. In addition, the enhanced expression of FRMD6 is associated with poor outcomes in patients with lung squamous cell carcinoma (n = 75, P = 0.0054) and lung adenocarcinoma (n = 94, P = 0.0330). Cell migration and proliferation in vitro and tumor formation in vivo are promoted by FRMD6 but are suppressed by the depletion of FRMD6. Mechanistically, FRMD6 interacts and colocalizes with mTOR and S6K, which are the key molecules of the mTOR signaling pathway. FRMD6 markedly enhances the interaction between mTOR and S6K, subsequently increasing the levels of endogenous pS6K and downstream pS6 in lung cancer cells. Furthermore, knocking out FRMD6 inhibits the activation of the mTOR signaling pathway in Frmd6^-/- gene KO MEFs and mice. Altogether, our results show that FRMD6 contributes to lung cancer progression by activating the mTOR signaling pathway.