Objective:To explore the role of tumor necrosis factor receptor-associated factor 4 (TRAF4) in promoting the abnormal activation of epidermal growth factor receptor (EGFR) and its effect on lung cancer cell proliferation, migration and invasion.Methods:Tumor tissues from patients who underwent lung adenocarcinoma resection at The First Affiliated Hospital of Second Military Medical University, from January 2015 to May 2017 were collected, and the expressions of TRAF4 and Ki-67 in lung cancer tissues were detected by immunohistochemistry, the mRNA levels of Cyclin D and Vimentin were detected by real-time fluorescence quantitative PCR (qRT-PCR). The effect of TRAF4 on the tumor growth ability of lung cancer A549 cells was investigated by the xenograft model, the effect of TRAF4 or EGFR on the tumor proliferation ability was detected by using cell counting kit 8 (CCK8) and BrdU assay, and the migration and invasion abilities of tumor cells were detected by Transwell assay. Different structural domain deletion expression vectors of TRAF4 and EGFR were constructed to transfect cells, and the interaction mode of TRAF4 and EGFR was investigated by immunoprecipitation assay.Results:The expression of TRAF4 in non-small cell lung cancer (NSCLC) tissues was positively correlated with the expressions of Ki-67, cyclin D, and vimentin ( r2: 0.438, 0.695, and 0.736, respectively, all P＜0.01). Immunohistochemical assay of tumor tissues from NSCLC patients showed that tissues with high expression of TRAF4 were also high in Ki-67. Patients with high TRAF4 expression (TRAF4 positivity >30%) had a shorter progression-free survival (PFS) time than that of patients with low TRAF4 expression (TRAF4 positivity ≤30%) (median PFS of 12 and 19 months, respectively; P=0.034). Traf4 -/- cells had a weakened proliferative capacity than traf4+/+ cells and formed tumors with smaller size ( P＜0.05). The expression level of Ki-67 in the tumor tissues formed by traf4 -/- cells [(45.6±8.7)%] was lower than that in the tumor tissues formed by traf4+/+ cells [(62.3±10.3)%, P=0.015], the mRNA levels of cyclin D (1.01±0.15) and vimentin (1.01±0.12) in the traf4 -/- cells were lower than those of the traf4+/+ cells (3.41±0.32 and 3.12±0.18, respectively, both P＜0.05).The western blot results showed that, with the elevated intracellular expression level of TRAF4, phosphorylation level of EGFR was significantly increased in both wild-type EGFR and activation mutant EGFR-expression cells. The capacities of proliferation, migration and invasion of A549 cells was weakened after EGFR knockdown (all P＜0.01). Immunoprecipitation experiments showed that TRAF4 binds to the peptide segment of the near-membrane region of EGFR through the TRAF structural domain, and the mutual binding between EGFR molecules was enhanced under TRAF4 overexpression conditions. Increasing TRAF4 expression promoted EGFR molecular phosphorylation and activation of downstream signaling. Conclusions:TRAF4 expression is elevated in NSCLC tissues and tumor cells, which promotes tumor proliferation, migration and invasion. TRAF4 directly binds to EGFR molecules, enhances its own phosphorylation and activates the downstream signaling pathway by promoting the interaction between EGFR molecules.

Objective:To explore the role of tumor necrosis factor receptor-associated factor 4 (TRAF4) in promoting the abnormal activation of epidermal growth factor receptor (EGFR) and its effect on lung cancer cell proliferation, migration and invasion.Methods:Tumor tissues from patients who underwent lung adenocarcinoma resection at The First Affiliated Hospital of Second Military Medical University, from January 2015 to May 2017 were collected, and the expressions of TRAF4 and Ki-67 in lung cancer tissues were detected by immunohistochemistry, the mRNA levels of Cyclin D and Vimentin were detected by real-time fluorescence quantitative PCR (qRT-PCR). The effect of TRAF4 on the tumor growth ability of lung cancer A549 cells was investigated by the xenograft model, the effect of TRAF4 or EGFR on the tumor proliferation ability was detected by using cell counting kit 8 (CCK8) and BrdU assay, and the migration and invasion abilities of tumor cells were detected by Transwell assay. Different structural domain deletion expression vectors of TRAF4 and EGFR were constructed to transfect cells, and the interaction mode of TRAF4 and EGFR was investigated by immunoprecipitation assay.Results:The expression of TRAF4 in non-small cell lung cancer (NSCLC) tissues was positively correlated with the expressions of Ki-67, cyclin D, and vimentin ( r2: 0.438, 0.695, and 0.736, respectively, all P＜0.01). Immunohistochemical assay of tumor tissues from NSCLC patients showed that tissues with high expression of TRAF4 were also high in Ki-67. Patients with high TRAF4 expression (TRAF4 positivity >30%) had a shorter progression-free survival (PFS) time than that of patients with low TRAF4 expression (TRAF4 positivity ≤30%) (median PFS of 12 and 19 months, respectively; P=0.034). Traf4 -/- cells had a weakened proliferative capacity than traf4+/+ cells and formed tumors with smaller size ( P＜0.05). The expression level of Ki-67 in the tumor tissues formed by traf4 -/- cells [(45.6±8.7)%] was lower than that in the tumor tissues formed by traf4+/+ cells [(62.3±10.3)%, P=0.015], the mRNA levels of cyclin D (1.01±0.15) and vimentin (1.01±0.12) in the traf4 -/- cells were lower than those of the traf4+/+ cells (3.41±0.32 and 3.12±0.18, respectively, both P＜0.05).The western blot results showed that, with the elevated intracellular expression level of TRAF4, phosphorylation level of EGFR was significantly increased in both wild-type EGFR and activation mutant EGFR-expression cells. The capacities of proliferation, migration and invasion of A549 cells was weakened after EGFR knockdown (all P＜0.01). Immunoprecipitation experiments showed that TRAF4 binds to the peptide segment of the near-membrane region of EGFR through the TRAF structural domain, and the mutual binding between EGFR molecules was enhanced under TRAF4 overexpression conditions. Increasing TRAF4 expression promoted EGFR molecular phosphorylation and activation of downstream signaling. Conclusions:TRAF4 expression is elevated in NSCLC tissues and tumor cells, which promotes tumor proliferation, migration and invasion. TRAF4 directly binds to EGFR molecules, enhances its own phosphorylation and activates the downstream signaling pathway by promoting the interaction between EGFR molecules.

Traumatic supraorbital fissure syndrome (TSOFS) is a symptom complex caused by nerve entrapment in the supraorbital fissure after skull base trauma. If the compressed cranial nerve in the supraorbital fissure is not decompressed surgically, ptosis, diplopia and eye movement disorder may exist for a long time and seriously affect the patients′ quality of life. Since its overall incidence is not high, it is not familiarized with the majority of neurosurgeons and some TSOFS may be complicated with skull base vascular injury. If the supraorbital fissure surgery is performed without treatment of vascular injury, it may cause massive hemorrhage, and disability and even life-threatening in severe cases. At present, there is no consensus or guideline on the diagnosis and treatment of TSOFS that can be referred to both domestically and internationally. To improve the understanding of TSOFS among clinical physicians and establish standardized diagnosis and treatment plans, the Skull Base Trauma Group of the Neurorepair Professional Committee of the Chinese Medical Doctor Association, Neurotrauma Group of the Neurosurgery Branch of the Chinese Medical Association, Neurotrauma Group of the Traumatology Branch of the Chinese Medical Association, and Editorial Committee of Chinese Journal of Trauma organized relevant experts to formulate Chinese expert consensus on the diagnosis and treatment of traumatic supraorbital fissure syndrome ( version 2024) based on evidence of evidence-based medicine and clinical experience of diagnosis and treatment. This consensus puts forward 12 recommendations on the diagnosis, classification, treatment, efficacy evaluation and follow-up of TSOFS, aiming to provide references for neurosurgeons from hospitals of all levels to standardize the diagnosis and treatment of TSOFS.

Objective To investigate the effects of B cell lymphoma factor-3(Bcl-3)on estrogen receptor α(ERα)signaling pathway and cell proliferation in breast cancer.Methods MCF-7 cells at the logarithmic growth stage were randomly divided into the siControl group and the siBcl-3#1 group.The cells were added with ControlsiRNA and siBcl-3#1siRNA,respectively,and transfected with LipofectamineTM 2000 for 5 hours.MCF-7 cells in the siControl group and siBcl-3#1 group were obtained after stable transfection for 48 hours.The expressions of Bcl-3,growth regulation by estrogen in breast cancer 1(GREB1),PDZ domain protein 1(PDZK1),and Erα target gene trefoil factor 1(TFF1)mRNA in MCF-7 cells were detected by quantitative real-time polymerase chain reaction.The expressions of Bcl-3 and ERα protein in MCF-7 cells were detected by Western blot assay,and the proliferation of MCF-7 cells was detected by cell counting kit-8.HEK293T cells were randomly divided into the Input group(positive control group),IP group(co-immunoprecipitation group),and IgG group(negative control group).After co-transfected with Bcl-3 and ERα plasmids for 48 hours,the cells were lysed with 100 μL co-immunoprecipitation(Co-IP)cell lysate on ice,and the supernatant was separated.In the Input group,90 μL supernatant was taken and added with 30 μL 4 × Loading buffer.In the IP group,450μL supernatant was taken and added with 40 μL protein A/G and 0.5 μL green fluorescent protein antibody.In the IgG group,450 μL supernatant was taken and added with 0.5 μL IgG and 30 μL protein A/G magnetic agarose beads.The binding effect of Bcl-3 and ERα was detected by Co-IP assay.MCF-7 cells at the logarithmic growth stage were selected,and the co-localization of Bcl-3 and ERα was detected by an immunofluorescence co-localization experiment.Results The relative expression levels of Bcl-3,GREB1,TFF1 and PDZK1 mRNA in MCF-7 cells in the si-Bcl-3#1 group were significantly lower than those in the siControl group(P＜0.05).The relative expression levels of Bcl-3 and ERα protein in MCF-7 cells in the si-Bcl-3#1 group were significantly lower than those in the siControl group(P＜0.05).There was no significant difference in the proliferation ability of MCF-7 cells between the si-Bcl-3#1 group and the siControl group at 0 h and 24 h after culture(P＞0.05).The proliferation ability of MCF-7 cells in the si-Bcl-3#1 group was significantly lower than that in the siControl group at 48 h and 72 h after culture(P＜0.05).The results of the Co-IP assay showed that there was interaction between Bcl-3 and ERα.The results of the immunofluorescence co-localization experiment showed that the co-localization of Bcl-3 and ERα existed.Conclusion Bcl-3 is highly expressed in breast cancer cells,which can enhance the conduction activity of the ERα signaling pathway and promote the proliferation of ERα positive breast cancer cells.

Objective To develop an auto-segmentation model based on no new U-net for delineating high-risk clinical target volume(HR-CTV)and organs-at-risk(OAR)in CT-guided brachytherapy of cervical cancer,and to explore its clinical value.Methods The CT images of 63 patients with locally advanced cervical cancer who had completed image-guided brachytherapy were collected.The HR-CTV and OAR including bladder,rectum and sigmoid colon were delineated manually by a senior oncologist,and the results were taken as the gold standard.The automatic and manual segmentation results were compared,and Dice similarity coefficient was used to evaluate HR-CTV and OAR auto-segmentation accuracies.Results The Dice similarity coefficients of HR-CTV,bladder,rectum,and sigmoid colon were 0.903±0.015,0.948±0.011,0.903±0.008,and 0.803±0.024,respectively.Conclusion The established model can realize the accurate segmentations of HR-CTV,bladder,rectum and sigmoid colon,but the oncologist still needs to scrupulously check the results.

Objective:To establish a new type of small intestinal organoids with injury-related regenerative capacity, and to simulate the process of intestinal injury, regeneration, and repair in vitro. Methods:The crypt structures of ileal mucosa from 6 to 8 weeks old, 18 to 24 g specific pathogen-free C57BL/6 mice were isolated. The ENR, ENR+ tumor necrosis factor-α(TNF-α) and 8C culture systems were designed to establish small intestinal organoids under conditions of intestinal homeostasis, inflammatory injury and injury-related regeneration, and the morphology of intestinal organoids were observed. The cell types and spatial arrangements of intestinal organoids, and the expression of genes clusterin( Clu), annexin A1( Anxa1), stem cell antigen-1( Sca1) and regenerating islet-derived protein 3-beta( Reg3 b) at protein levels were detected by immunofluorescence staining. The expression of genes Clu, Anxa1, Sca1 and Reg3 b at mRNA levels were detected by quantitative real-time polymerase chain reaction (qRT-PCR). Independent sample- t test was used for statistical analysis. Results:In ENR and 8C culture system, both intestinal organoids contained intestinal stem cells, goblet cells, Paneth cells and intestinal endocrine cells, and the spatial arrangement of cells was similar to the intestinal epithelium. In the 8C culture system, the amplification capacity of the new small intestinal organoids was significantly enhanced, the growth rate was faster, and the structure was larger and more complex than those of small intestinal organoids in ENR and ENR+ TNF-α culture systems. The results of qRT-PCR showed that, the relative mRNA expression levels of novel small intestinal organoid regeneration genes Clu, Anxa1, and Sca1 in the 8C culture system were higher than those in the ENR and ENR+ TNF-α culture systems (0.68±0.31 vs.0.20±0.07 and 0.36±0.19, 0.48±0.13 vs. 0.07±0.02 and 0.18±0.11, 0.56±0.20 vs. 0.02±0.01 and 0.08±0.04), and the differences were statistically significant ( t=4.82 and 2.77, 8.62 and 4.89, and 8.58 and 7.50; all P<0.05). The results of immunofluorescence staining indicated that, the expression levels of novel small intestinal organoid regeneration genes Clu, Anxa1, Sca1 and Reg3 b at protein level in the 8C culture system were higher than those in the ENR and ENR+ TNF-α culture systems (31.62±1.69 vs. 9.73±2.39 and 15.11±2.16, 42.65±1.85 vs. 19.70±1.18 and 24.97±2.82, 63.80±2.73 vs. 37.10±1.59 and 43.27±2.53, 53.26±1.84 vs. 27.75±3.78 and 33.16±3.50), and the differences were statistically significant( t=12.95 and 10.41, 18.13 and 9.09, 14.63 and 9.56, and 10.51 and 8.80; all P<0.001). Conclusion:The small intestinal organoids established in the novel culture system have the characteristics of injury-related regeneration, and provide a novel in vitro model for studying the regeneration of epithelial tissues and organs.

Objective:To investigate the value of nomogram predictive model established by the risk factors of upper extremity venous thrombosis risk associated with peripherally venous inserted central catheter (PICC) in cancer patients.Methods:A total of 1 032 patients who underwent PICC insertion between January 2016 and March 2017 in Zhejiang Cancer Hospital were selected by using prospective cohort study and convenience sampling. Risk factors of upper extremity venous thrombosis risk associated with PICC in cancer patients were evaluated by using Cox regression model. The nomogram predictive model of upper extremity venous thrombosis risk associated with PICC insertion was constructed. Bootstrap method was used to complete the inside check, and figure calibration was used to verify the nomogram.Results:A multivariate Cox regression analysis showed that trombosis history ( HR = 27.82, 95% CI 8.17-94.88, P < 0.01) and hyperlipidemia ( HR = 3.01, 95% CI 1.31-6.93, P = 0.009) were independent risk factors for upper extremity venous thrombosis associated with PICC. The nomogram model C-index was 0.71 (95% CI 0.63-0.80) based on the above risk factors, which indicated that the nomogram had a good differentiation. The calibration curve for predicting the probability of upper extremity venous thrombosis risk associated with PICC within one week, two weeks and one month deviated slightly from the standard curve, suggesting that the model might overestimate the risk of upper extremity venous thrombosis associated with PICC in cancer patients. Conclusions:The nomogram model has a good predictive value and strong operability, which can be used to predict the probability of upper extremity venous thrombosis associated with PICC in cancer patients after PICC insertion. It can provide a reference for identifying the high-risk cancer patients and formulating proper therapeutic strategies.

Objective:To explore the incidence and risk factors of peripherally inserted central catheter (PICC) related upper extremity venous thrombosis (UEVT) in patients with head and neck neoplasm so as to provide a basis for preventing thrombosis.Methods:This study used the design of prospective cohort study. From January 2016 to March 2018, UEVT follow-up examination by B ultrasound was carried out for 1 137 head and neck neoplasm patients with PICC selected by convenience sampling. Single factor and multivariate Cox regression were used to determine the risk factors of PICC related UEVT.Results:There were 3.6% (41/1 137) of patients with PICC related UEVT. Multivariate Cox regression showed that the independent risk factors of PICC related UEVT included the older patients ( RR=1.04, 95% CI: 1.01-1.07, P=0.013) , being with a history of PICC catheterization ( RR=3.22, 95% CI: 1.53-6.77, P=0.002) and high frequency of catheter delivery ( RR=1.98, 95% CI: 1.30-3.00, P=0.001) . Conclusions:Patients with head and neck neoplasm have the low incidence of PICC related UEVT. The independent risk factors of PICC related UEVT in patients with head and neck neoplasm include the older ages, history of PICC catheterization and high frequency of catheter delivery. Positive intervention should be carried out for those patients which may reduce the incidence of PICC related thrombosis.

Temporomandibular disorders (TMD) is one of the most common diseases in the orofacial region. The occurrence, development and outcome of TMD are affected by many factors. Among various risk factors, the psychological factors, especially anxiety, depression and somatic symptoms, are getting more and more attention in the etiology, diagnosis and treatment of TMD. Psychological factors are associated with the occurrence of TMD, and the accurate diagnostic criteria is conducive to the assessment of the patient′s psychological state. If necessary, an appropriate psychological treatment according to a patient′s psychological status can effectively improve the effect of clinical treatment. This article, based on domestic and international literatures, reviews the research progress of the correlation between the psychological factors and the etiology, diagnosis and treatment of TMD, in order to provide new ideas for clinicians to diagnose and treat TMD.

Objective: To understand the etiology, genotype and molecular characteristics of acute viral gastroenteritis in Quanzhou from 2014 to 2017. Methods: Specimens from 15 outbreaks of acute viral gastroenteritis in Quanzhou area from 2014 to 2017 were collected and real-time fluorescence quantitative PCR was used to detect norovirus GI and GII, sapovirus, astrovirus and rotavirus, and the result were statistically analyzed. Furthermore, specimens positive for norovirus was further subjected to the amplification and sequencing of polymerase and VP1 genes of norovirus, and sequences were analyzed using DNAstar and MEGA7.0 software. Results: In this study, 96 specimens from 15 outbreaks of acute viral gastroenteritis were collected, and norovirus was detected in 30 specimens with a positive rate of 31.25%, among which 23 specimens were genotype GII and 7 specimens genotype GI. Meanwhile, 10 specimens were randomly selected for nucleic acid sequence analysis. The result showed that 9 of them were GII.P16/GII.2 and 1 was GI.6. The phylogenetic analysis showed that the new recombinant norovirus subtype GII.P16/GII.2 was highly homologous to the same subtype detected in outbreaks home and abroad recently. Conclusions The main pathogens caused the outbreak of acute viral gastroenteritis in Quanzhou from 2014 to 2017 were norovirus belonging to subtype GII.P16/GII.2 and subtype GI.6, and subtype GII.P16/GII.2 was the predominant strain which was found for the first time in Quanzhou.