Drug resistance mechanisms and treatment strategies mediated by Ubiquitin-Specific Proteases (USPs) in cancers: new directions and therapeutic options.

Drug resistance represents a significant obstacle in cancer treatment, underscoring the need for the discovery of novel therapeutic targets. Ubiquitin-specific proteases (USPs), a subclass of deubiquitinating enzymes, play a pivotal role in protein deubiquitination. As scientific research advances, USPs have been recognized as key regulators of drug resistance across a spectrum of treatment modalities, including chemotherapy, targeted therapy, immunotherapy, and radiotherapy. This comprehensive review examines the complex relationship between USPs and drug resistance mechanisms, focusing on specific treatment strategies and highlighting the influence of USPs on DNA damage repair, apoptosis, characteristics of cancer stem cells, immune evasion, and other crucial biological functions. Additionally, the review highlights the potential clinical significance of USP inhibitors as a means to counter drug resistance in cancer treatment. By inhibiting particular USP, cancer cells can become more susceptible to a variety of anti-cancer drugs. The integration of USP inhibitors with current anti-cancer therapies offers a promising strategy to circumvent drug resistance. Therefore, this review emphasizes the importance of USPs as viable therapeutic targets and offers insight into fruitful directions for future research and drug development. Targeting USPs presents an effective method to combat drug resistance across various cancer types, leading to enhanced treatment strategies and better patient outcomes.

Cellular functions, molecular signalings and therapeutic applications: Translational potential of deubiquitylating enzyme USP9X as a drug target in cancer treatment.

Protein ubiquitination, one of the most significant post-translational modifications, plays an important role in controlling the proteins activity in diverse cellular processes. The reversible process of protein ubiquitination, known as deubiquitination, has emerged as a critical mechanism for maintaining cellular homeostasis. The deubiquitinases (DUBs), which participate in deubiquitination process are increasingly recognized as potential candidates for drug discovery. Among these DUBs, ubiquitin-specific protease 9× (USP9X), a highly conserved member of the USP family, exhibits versatile functions in various cellular processes, including the regulation of cell cycle, protein endocytosis, apoptosis, cell polarity, immunological microenvironment, and stem cell characteristics. The dysregulation and abnormal activities of USP9X are influenced by intricate cellular signaling pathway crosstalk and upstream non-coding RNAs. The complex expression patterns and controversial clinical significance of USP9X in cancers suggest its potential as a prognostic biomarker. Furthermore, USP9X inhibitors has shown promising antitumor activity and holds the potential to overcome therapeutic resistance in preclinical models. However, a comprehensive summary of the role and molecular functions of USP9X in cancer progression is currently lacking. In this review, we provide a comprehensive delineation of USP9X participation in numerous critical cellular processes, complicated signaling pathways within the tumor microenvironment, and its potential translational applications to combat therapeutic resistance. By systematically summarizing the updated molecular mechanisms of USP9X in cancer biology, this review aims to contribute to the advancement of cancer therapeutics and provide essential insights for specialists and clinicians in the development of improved cancer treatment strategies.

MAGP2 promotes osteogenic differentiation during fracture healing through its crosstalk with the ß-catenin pathway.

Osteogenic differentiation is important for fracture healing. Microfibrial-associated glycoprotein 2 (MAGP2) is found to function as a proangiogenic regulator in bone formation; however, its role in osteogenic differentiation during bone repair is not clear. Here, a mouse model of critical-sized femur fracture was constructed, and the adenovirus expressing MAGP2 was delivered into the fracture site. Mice with MAGP2 overexpression exhibited increased bone mineral density and bone volume fraction (BV/TV) at Day 14 postfracture. Within 7 days postfracture, overexpression of MAGP2 increased collagen I and II expression at the fracture callus, with increasing chondrogenesis. MAGP2 inhibited collagen II level but elevated collagen I by 14 days following fracture, accompanied by increased endochondral bone formation. In mouse osteoblast precursor MC3T3-E1 cells, MAGP2 treatment elevated the expression of osteoblastic factors (osterix, BGLAP and collagen I) and enhanced ALP activity and mineralization through activating ß-catenin signaling after osteogenic induction. Besides, MAGP2 could interact with lipoprotein receptor-related protein 5 (LRP5) and upregulated its expression. Promotion of osteogenic differentiation and ß-catenin activation mediated by MAGP2 was partially reversed by LRP5 knockdown. Interestingly, ß-catenin/transcription factor 4 (TCF4) increased MAGP2 expression probably by binding to MAGP2 promoter. These findings suggest that MAGP2 may interact with ß-catenin/TCF4 to enhance ß-catenin/TCF4's function and activate LRP5-activated ß-catenin signaling pathway, thus promoting osteogenic differentiation for fracture repair. mRNA sequencing identified the potential targets of MAGP2, providing novel insights into MAGP2 function and the directions for future research.

[Research progress of pubic symphysis diastasis].

Objective: To review the research progress of pubic symphysis diastasis and provide effective reference for orthopedic surgeons in the diagnosis and treatment of pubic symphysis diastasis. Methods: The anatomy, injury mechanism, treatment, and other aspects of pubic symphysis diastasis were summarized and analyzed by reviewing the relevant research literature at domestically and internationally in recent years. Results: The incidence of pubic symphysis diastasis is high in pelvic fractures, which is caused by the injury of the ligaments and fibrocartilage disc around the pubic symphysis by external force. The treatment plan should be individualized according to the pelvic stability and the needs of patients, aiming to restore the stability and integrity of the pelvis and improve the quality of life of patients after surgery. Conclusion: At present, the research on pubic symphysis diastasis still needs to be improved. In the future, high-quality, multi-center, and large-sample studies are of great significance for the selection of treatment methods and the evaluation of effectiveness for patients with pubic symphysis diastasis.

Disordered Convolution Region of P(VDF-TrFE) Piezoelectric Nanoparticles: The Core of Sono-Piezo Dynamic Therapy.

The recent focus on P(VDF-TrFE) material in biomedical engineering stems from its outstanding mechanical properties and biocompatibility. However, its application in sono-piezo dynamic therapy (SPDT) has been relatively unexplored. In this study, we developed composite piezoelectric nanoparticles (rPGd NPs@RGD) based on recrystallized P(VDF-TrFE) particles, which offer dual capabilities of MRI imaging and targeted treatment for brain gliomas. SEM observations of P(VDF-TrFE) particles in the disordered convolution region (DCR) revealed recrystallization, representing the polymer chain structure and particle polarity. In comparison to nonrecrystallized nanoparticles, rPGd NPs@RGD exhibited remarkable stability and biocompatibility. Under ultrasound excitation, they generated significantly higher levels of reactive oxygen species, effectively inhibiting tumor cell proliferation, invasion, and migration. rPGd NPs@RGD demonstrated excellent MRI imaging capabilities and antitumor activity in U87 tumor-bearing mice. This study highlights the remarkable SPDT abilities of the developed nanoparticles, attributed to the microscopic morphological changes in the DCR that increase the nanoparticle's polarity and thus boost its potential for SPDT. This research opens new possibilities for utilizing P(VDF-TrFE) materials in advanced biomedical applications.

Direct Knockdown of PDZ and LIM Domain 1 Using an Adenoviral Delivery System Accelerates Osteogenesis and Fracture Healing in Mice.

Substantial advances have been made in understanding the role of partial PDZ and LIM domain family's proteins in skeletal-related diseases. Yet, little is known about the effect of PDZ and LIM Domain 1 (Pdlim1) on osteogenesis and fracture repair. This study aimed to investigate whether direct gene delivery using an adenovirus vector carrying Pdlim1 (Ad-oePdlim1) or encoding shRNA-Pdlim1 (Ad-shPdlim1) could affect the osteogenic activity of preosteoblastic MC3T3-E1 cells in vitro, and influence the fracture healing of mice in vivo. We found that Ad-shPdlim1 transfection contributed to the calcified nodule formation in MC3T3-E1 cells. Downregulation of Pdlim1 enhanced the alkaline phosphatase activity and increased the expression of osteogenic markers (Runt-related transcription factor 2 [Runx2], collagen type I alpha 1 chain [Col1A1], osteocalcin [OCN], and osteopontin [OPN]). Further analysis indicated that Pdlim1 knockdown could activate ß-catenin signaling, as evidenced by the accumulation of ß-catenin in the nucleus and the increase levels of downstream regulators such as Lef1/Tcf7, axis inhibition protein 2, cyclin D1, and SRY-box transcription factor 9. By contrast, Pdlim1 overexpression resulted in inhibition of the osteogenic activity of MC3T3-E1 cells. In vivo, at day 3 after fracture,Ad-shPdlim1 adenovirus particles were injected into the fracture site of the femur of mice, and the process of fracture healing was evaluated by X-ray, micro-computed tomography and histological examination. Local injection of Ad-shPdlim1 promoted the early cartilage callus formation, restored bone mineral density, and accelerated cartilaginous ossification, with the upregulation of osteogenic gene (Runx2, Col1A1, OCN, and OPN) expression and activation of ß-catenin signaling. Thus, we concluded that inhibition of Pdlim1 contributed to osteogenesis and fracture healing by activating the ß-catenin signaling pathway.

Microfibril-Associated Glycoprotein-2 Promoted Fracture Healing via Integrin αvß3/PTK2/AKT Signaling.

Fracture healing is a complex physiological process in which angiogenesis plays an essential role. Microfibril-associated glycoprotein-2 (MAGP2) has been reported to possess a proangiogenic activity via integrin αvß3, yet its role in bone repair is unexplored. In this study, a critical-sized femoral defect (2 mm) was created in mice, followed by the delivery of an adenovirus-based MAGP2 overexpression vector or its negative control at the fracture site. At days 7, 14, 21, and 28 postfracture, bone fracture healing was evaluated by radiography, micro-computed tomography, and histopathologic analysis. Adenovirus-based MAGP2 overexpression vector-treated mice exhibited increased bone mineral density and bone volume fraction. MAGP2 overexpression contributed to an advanced stage of endochondral ossification and induced cartilage callus into the bony callus. Further analysis indicated that MAGP2 was associated with enhanced angiogenesis, as evidenced by marked MAGP2 and integrin αvß3 costaining and increased endothelial cell markers such as endomucin and CD31 levls, as well as elevated phosphorylation of protein tyrosine kinase 2 (PTK2) and AKT serine/threonine kinase 1 (AKT) in the callus. In vitro, recombinant human MAGP2 treatment enhanced the viability, migration, and tube formation ability of human microvascular endothelial cells, which was partially reversed by integrin αvß3 inhibition or MK-2206, a specific AKT inhibitor. Inhibition of integrin αvß3 abolished MAGP2-induced PTK2 and AKT activation. Taken together, our data provide the first evidence that MAGP2 promotes angiogenesis and bone formation by activating the integrin αvß3/PTK2/AKT signaling pathway.

Transcranial sonography with clinical and demographic characteristics to predict cognitive impairment in PD: a longitudinal study.

BACKGROUND: Parkinson's disease (PD) is a neurodegenerative disease and is clinically characterized by a series of motor symptoms (MS) and nonmotor symptoms (NMS). NMS often appear before MS, while cognitive impairment mostly occurs within a few years after the diagnosis of PD. Therefore, we aimed to predict the risk factors for cognitive impairment (CI) in PD patients based on transcranial sonography, clinical symptoms, and demographic characteristics. METHODS: Based on the occurrence time of CI, a total of 172 PD patients were divided into non-CI (N-CI, n = 48), CI at the first treatment (F-CI, n = 58), and CI at the last treatment (L-CI, n = 66) groups. Clinical data (including MS and NMS) and ultrasonic data of all patients at the first treatment and the last treatment were collected retrospectively. Independent samples t tests were used to compare continuous data, and chi-square tests were used to compare categorical data. The risk factors for CI and Parkinson's disease dementia were identified by logistic regression analysis, and an ROC curve was established to explore the diagnostic efficacy. RESULTS: 1) The age of onset, first treatment and smoking history of CI patients were significantly different from those of N-CI patients. When age of first treatment ≥61 years was considered the boundary value to diagnose CI, the sensitivity and specificity were 77.40 and 66.70%, respectively. 2) The severity of depression was significantly different between F-CI and N-CI patients at the first treatment, while the cumulative and new or aggravated memory deficit was significantly different between the L-CI and N-CI patients at the last treatment. 3) There was a significant difference in TCS grading between the first and last treatment in L-CI patients. 4) Depression, sexual dysfunction, and olfactory dysfunction in NMS were independent risk factors for CI during the last treatment. 5) The sensitivity and specificity of predicting CI in PD patients were 81.80 and 64.60%, respectively. CONCLUSIONS: PD patients with CI were older, and most of them had a history of smoking. Furthermore, there was good diagnostic efficiency for predicting CI in PD via TCS combined with clinical characteristics (especially NMS).

Ultrasound-guided thermal ablation for papillary thyroid microcarcinoma: A systematic review.

OBJECTIVE: Thyroidectomy is the first-line treatment for papillary thyroid microcarcinoma (PTMC), but often involves aggressive overtreatment. Thermal ablation (TA) has been gradually used for the treatment of recurrent PTMC. However, it is not recommended for the treatment of primary PTMC according to the Korean and Italian guidelines. Therefore, this systematic review aimed to analyse the indications, efficacy, and safety of TA in the treatment of PTMC. DESIGN: Systematic review. PATIENTS AND MEASUREMENTS: A search strategy was developed in accordance with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses statement. A total of 27 articles were included in this study until January 2022. RESULTS: According to current guidelines and studies, we divided the indications of TA for PTMC into six primary and three secondary indications. Laser ablation (LA) has the advantages of a small needle, accurate output energy and precision ablation, and it is safe to important organs around the lesion. The patients recover quickly after radiofrequency ablation (RFA), with no major complications, recurrence, or lymph node metastasis. The volume reduction rate after RFA was the highest, followed by microwave ablation and LA, and the improvement in patient quality of life after TA was significantly better than after thyroidectomy. CONCLUSIONS: TA is an effective alternative method for surgery in the treatment of low-risk PTMC and has the advantages of being minimally invasive, economical, having less bleeding and having a high postoperative quality of life.

Diagnostic performance of simplified TI-RADS for malignant thyroid nodules: comparison with 2017 ACR-TI-RADS and 2020 C-TI-RADS.

BACKGROUND: The aim of this study is to propose a new TI-RADS and compare it with the American College of Radiology (2017 ACR)-TI-RADS and the 2020 Chinese (2020 C)-TI-RADS. METHODS: A retrospective analysis of 749 thyroid nodules was performed. Based on the calculated odds ratio of ultrasonic signs between benign and malignant nodules, a new thyroid nodule score and malignancy rate were calculated. A receiver operating characteristic curve was drawn to analyze the new system's effectiveness in the differential diagnosis of benign and malignant thyroid nodules and was compared with the 2020 C-TI-RADS and 2017 ACR-TI-RADS. Five ultrasound physicians with different qualifications graded another 123 thyroid nodules according to the 2017ACR-TI-RADS, 2020 C-TI-RADS, and the newly proposed TI-RADS. Intergroup and intragroup consistency was evaluated using the Kappa test and intraclass correlation coefficient (ICC) test. RESULTS: 1) The new thyroid nodule score was divided into 0, 1, 2, 3, 4, and 5 points, with malignancy rates of 1.52%, 7.69%, 38.24%, 76.00%, 90.75%, and 93.75%, respectively. Using 3 points as the cutoff value to diagnose benign and malignant thyroid nodules, the sensitivity and specificity were 94.03% and 67.39%, respectively, which were higher than those of the 2017 ACR-TI-RADS and 2020 C-TI-RADS. The simplified TI-RADS, namely, sTI-RADS, was established as follows: sTI-RADS 3 (0 points), malignancy rate < 2%; sTI-RADS 4a (1 point), malignancy rate 2-10%; sTI-RADS 4b (2 points), malignancy rate 10-50%; sTI-RADS 4 (3 points), malignancy rate 50-90%; and sTI-RADS 5 (4 and 5 points), malignancy rate > 90%. 2) Five ultrasound doctors graded thyroid nodules by the 2017 ACR-TI-RADS, 2020C-TI-RADS and sTI-RADS. Intragroup consistency was good among all tests; ICC were 0.86 (0.82-0.90), 0.84 (0.78-0.88), and 0.88 (0.84-0.91), respectively, while only sTI-RADS had good intergroup consistency. CONCLUSION: In summary, we proposed a new TI-RADS, namely, sTI-RADS, which was obtained using a simple assignment method with higher specificity, accuracy, positive predictive value, and Youden index than the 2017 ACR-TI-RADS and 2020 C-TI-RADS.

Noninvasive assessment of intracranial pressure using subharmonic-aided pressure estimation: An experimental study in canines.

BACKGROUND: Intracranial hypertension is a common clinicopathological syndrome in neurosurgery, and a timely understanding of the intracranial pressure (ICP) may help guide clinical treatment. We aimed to investigate the correlation between subharmonic contrast-enhanced ultrasound (SHCEUS) parameters and ICP in experimental canines. METHODS: A dynamic model of ICP change from 11 mm Hg to 50 mm Hg was established in experimental canines by placing a latex balloon into the epidural space and injecting saline into the balloon. In addition, a pressure sensor was placed in the brain parenchyma to record the changes in ICP. When the ICP stabilized after each increase, subharmonic-aided pressure estimation (SHAPE) technology was performed to obtain the SHCEUS parameters, including the basal venous and adjacent intracranial arterial subharmonic amplitude and SHAPE gradient (subharmonic amplitude in the intracranial artery minus that in the basal vein). The correlation between these parameters and ICP was analyzed. RESULTS: The subharmonic amplitude of the basal vein was negatively correlated with the ICP (r = -0.798), and the SHAPE gradient was positively correlated with the ICP (r = 0.628). According to the guidelines for ICP monitoring in patients with traumatic brain injury, we defined 20 mm Hg, 25 mm Hg, and 30 mm Hg as the cutoff ICP levels. The area under the receiver operating characteristic curve of the basal venous subharmonic amplitude for diagnosing intracranial hypertension ≥20 mm Hg, ≥25 mm Hg, and ≥30 mm Hg was 0.867 (95% confidence interval [CI], 0.750-0.943), 0.884 (95% CI, 0.770-0.954), and 0.875 (95% CI, 0.759-0.948), respectively. The area under the receiver operating characteristic curve of the SHAPE gradient for diagnosing intracranial hypertension ≥20 mm Hg, ≥25 mm Hg, and ≥30 mm Hg was 0.839 (95% CI, 0.716-0.924), 0.842 (95% CI, 0.720-0.926), and 0.794 (95% CI, 0.665-0.890), respectively. CONCLUSION: SHCEUS parameters are correlated with ICP. The SHAPE technique can assist in evaluating ICP changes in canines, which provides a new idea and method for evaluating ICP.

Efficacy and Safety of Thermal Ablation for Treating Lymph Node Metastasis From Papillary Thyroid Carcinoma: A Systematic Review and Meta-Analysis.

Objective: To evaluate the efficacy and safety of thermal ablation, including radiofrequency ablation (RFA), microwave ablation (MVA), and laser ablation (LA), for treating lymph node metastasis (LNM) from papillary thyroid carcinoma (PTC). Design and Methods: PubMed and EMBASE were searched for studies reporting the efficacy and safety of thermal ablation for treating LNM in PTC. After selecting the relevant literature (including 11 papers, 208 patients, 412 lymph nodes), the QUADAS-2 tool was used to evaluate its quality. Then, both the fixed-effects and random-effects models combined with subgroup analysis were used to calculate data on volume changes in metastatic lymph nodes and changes in serum thyroglobulin (Tg) levels. We pooled the proportion of major and overall complication rates and complete disappearance rates and used subgroup forest plots and funnel plots for visual representation. Because of publication bias, we also performed a trim-and-filled model for correction. The rate of recurrence and distant metastasis with ablated details were pooled. Results: In the 11 articles (208 patients and 412 diseased lymph nodes), all thermal ablation methods showed effectiveness in reducing lymph node volume (P = 0.02) and serum Tg levels (P < 0.01) which showed no between-group difference. The pooled proportion of major complications was 0%(95% CI: -0.14; 0.15, P = 1) and the overall complication rate was 5% (95% CI: -0.09; 0.20, P = 1), which revealed no significant difference among modalities. The pooled proportion of the complete disappearance rate was 82% (95% CI: 0.43; 0.96, P < 0.01) and the data with statistical significance which contains RFA and LA showed complete disappearance rate was 59% and 81% respectively. Conclusion: All thermal ablation methods, including RFA, MWA, and LA, were effective and safe for treating LNM in PTC and were especially suitable for nonsurgical patients. Besides, subgroup analysis showed no significant difference, except for LA is better than RFA in complete disappearance rate.

Ultrasound-guided thermal ablation for hyperparathyroidism: current status and prospects.

BACKGROUND: Hyperparathyroidism (HPT) is classified into primary HPT (PHPT), secondary HPT (SHPT), tertiary HPT (THPT), and pseudohyperparathyroidism. Parathyroid surgery is generally reserved for patients with symptomatic PHPT and asymptomatic patients who meet the surgical guideline criteria. However, the risk of complications and mortality after parathyroid gland surgery increases with increasing patient age. AIM: This study aimed to review existing research on laser ablation, radiofrequency ablation, microwave ablation, and high-intensity focused ultrasound in the treatment of HPT and analyze its application prospects. CONCLUSIONS: Thermal ablation is a good alternative treatment for patients with parathyroid hyperplasia who do not meet the criteria or decline surgery. Being a type of minimally invasive treatment, ultrasound-guided thermal ablation has the advantages of easy operation, rapid recovery, and reusability and is used widely.

Effects of the p16/cyclin D1/CDK4/Rb/E2F1 pathway on aberrant lung fibroblast proliferation in neonatal rats exposed to hyperoxia.

p16INK4a (p16) inhibits the vital G1 to S phase transition during cell cycle progression through the p16/cyclin D1/CDK4/retinoblastoma(Rb)/E2F1 pathway. Hyperoxia can suppress the G1/S checkpoint and induce more lung fibroblasts (LFs) to transition from the G1 phase to the S phase and undergo cell proliferation. The present study investigated the rate of p16 gene promoter methylation and the protein expression levels of p16, cyclin D1, CDK4, Rb and E2F1 in LFs from the lungs of rats exposed to hyperoxia and normoxia on postnatal days 3, 7 and 14. In the hyperoxia-exposed group, the methylation rate was 50 and 80% on days 7 and 14, respectively. Cyclin D1 and CDK4 overexpression was associated with p16 loss and Rb inactivation by phosphorylation. Rb phosphorylation induced E2F1 release in the G1 phase, which promoted cell proliferation. No methylation was observed in the normoxia-exposed group. These observations suggested that p16 loss may stimulate aberrant LF proliferation via the p16/cyclin D1/CDK4/Rb/E2F1 pathway.

Grass carp (Ctenopharyngodon idella) stefin A: Systematic research on its cloning, expression, characterization and tissue distribution.

To fully understand the properties of piscine stefins (family I cystatins), the 294-bp stefinA gene from grass carp (Ctenopharyngodon idella, Ci) was cloned and expressed in E. coli BL21 (DE3). After purification by Ni2+-NTA agarose affinity chromatography, the CiStefin A protein was tested to have a molecular weight of 11.48 kDa and an isoelectric point of 8.7. The typical motif of the cystatins superfamily was characterized from CiStefin A (QVVQG). CiStefin A specifically inhibited the activity of papain and cathepsin B/L. The Ki value of CiStefin A against papain was 6.5 × 10-11 M. CiStefin A showed excellent heat and acid-base tolerance. StefinA gene transcription occurred in all tested tissues of grass carp, with the highest level in the hepatopancreas. Immunolocalization staining with an anti-CiStefinA antibody revealed the CiStefinA protein distribution in all tested tissues at various levels. Overall, these results clarified the physical and biochemical properties of grass carp stefin A.

XBP1 negatively regulates CENPF expression via recruiting ATF6α to the promoter during ER stress.

BACKGROUND: Centromere protein F (CENPF) is a key component of the kinetochore complex involved in mitosis, cell differentiation and cellular response to stresses. However, the alteration of CENPF in response to endoplasmic reticulum (ER) stress has not been well described. In the present study, we investigate CENPF regulation in response to ER stress. METHODS: Quantitative real-time polymerase chain reaction and western blotting were used to determine CENPF expression under ER stress. Luciferase activity analysis was performed to investigate the promoter regions contributing to CENPF transcription in response to TG. Chromatin immunoprecipitation (ChIP) and ChIP Re-IP assays were used to determine if X-box binding protein 1 (XBP1) and/or activating transcription factor 6α (ATF6α) bind in the CENPF promoter region. Cell apoptosis and proliferation were analyzed using TUNEL, cell growth and clonogenic assays. RESULTS: CENPF expression is dramatically reduced under ER stress induced by thapsigargin (TG), brefeldin A (BFA), or tunicamycin (TM) and this downregulation of CENPF expression was dependent on XBP1 and ATF6α. Luciferase activity analysis of the truncated CENPF promoter indicates that regions from bases - 679 to - 488 and from - 241 to - 78 in the CENPF promoter were sensitive to TG treatment. Additionally, ChIP and ChIP Re-IP assays reveal that XBP1 and ATF6α were assembled on the same regions of CENPF promoter. Notably, we identify two XBP1 binding sequences at positions - 567 and - 192, to which XBP1 binding was enhanced by TG. Finally, CENPF overexpression inhibits cell apoptosis and promotes cell proliferation in response to ER stress. CONCLUSION: In summary, these results demonstrate that ER stress plays a crucial role in CENPF expression, and XBP1 may up-regulate DNA-binding affinities after TG treatment to the promoter of CENPF. These findings may contribute to the understanding of the molecular mechanism of CENPF regulation.

Greater Omentum Imaging-Reporting and Data System: establishing the grade of benign and malignant lesions of the greater omentum using ultrasonography.

OBJECTIVE: To establish Greater Omentum Imaging-Reporting and Data System (GOI-RADS) to evaluate the possibility of omental diseases being malignant. METHOD: A retrospective analysis was made of 883 patients who had undergone biopsy of the greater omentum in our center from October 2009 to October 2019. Twelve parameters of ultrasonographic images were evaluated, and the odds ratio of each group calculated. We assigned scores for the direct signs (omental echo, omental structure, and omental nodules) and indirect signs (separation of ascites, echo of ascites, mesenteric lymph nodes, and thickening of parietal peritoneum) of omental lesions. We created an omental score (OS) for each patient and receiver operating characteristic (ROC) curve to analyze its effectiveness in the differential diagnosis of benign and malignant omental diseases. RESULTS: The OS was divided into ≤5, 6, 7, 8, 9, 10, 11, 12, 13, and ≥ 14 points, and the malignant rate was 0, 1.85, 5.56, 30.36, 37.25, 87.72, 96.72, 98.28, 99.08, and 100%, respectively. The area under the ROC curve (AUC) was 0.976. When taking 10 points as the cutoff value to diagnose benign and malignant omental diseases, the sensitivity and specificity was 93.85 and 98.21%, respectively. A grading system was established: grade 1: omental score ≤ 5, malignant rate 0%; grade 2: omental score 6-7, malignant rate ≤ 5.56%; grade 3: omental score 8--9, malignant rate ≤ 37.25%; grade 4: omental score ≥ 10, malignant rate ≥ 87.72. CONCLUSION: GOI-RADS had high sensitivity and specificity in the differential diagnosis of benign and malignant omental lesions. We believe that GOI-RADS will aid the diagnosis of omental diseases based on objective and accurate interpretation of ultrasound features, and also to promote the ultrasonography of omental diseases in clinical application.

Activating transcription factor 6 (ATF6) negatively regulates Polo-like kinase 4 expression via recruiting C/EBPß to the upstream-promoter during ER stress.

Polo-like kinase 4 (PLK4) is a member of the serine/threonine protein kinase family involved in cell-cycle regulation and cellular response to stresses. However, the alteration of PLK4 in response to endoplasmic reticulum (ER) stress has not been well described. In the present study, we focused on the regulation of PLK4 regulation in response to ER stress. PLK4 expression was dramatically reduced under ER stress induced by brefeldin A (BFA), tunicamycin (TM), or thapsigargin (TG) and down regulation of PLK4 expression was dependent on activating transcription factor 6 (ATF6) and CCAAT/enhancer-binding protein ß (C/EBPß). Luciferase activity analysis of the truncated PLK4 promoter indicated that region from -1343 to -1250 of the PLK4 promoter was sensitive to BFA or TG. Additionally, ChIP and ChIP Re-IP assays showed that ATF6 and C/EBPß were assembled on the same region of Plk4 promoter. Notably, we identified one C/EBPß responsive element at position -1284, to which ATF6 or C/EBPß binding was enhanced by BFA or TG under in vitro and in vivo conditions. Finally, overexpression of PLK4 inhibits apoptosis and promotes cell proliferation in response to ER stress. In summary, these results demonstrated that ER stress plays a crucial role in PLK4 expression. ATF6 may upregulate DNA-binding affinities after BFA treatment, via recruiting C/EBPß to the upstream promoter of PLK4. These findings may contribute to the understanding of the molecular mechanism of PLK4 regulation.

Renal cell carcinoma: predicting RUNX3 methylation level and its consequences on survival with CT features.

PURPOSE: To investigate associations between CT imaging features, RUNX3 methylation level, and survival in clear cell renal cell carcinoma (ccRCC). MATERIALS AND METHODS: Patients were divided into high RUNX3 methylation and low RUNX3 methylation groups according to RUNX3 methylation levels (the threshold was identified by using X-tile). The CT scanning data from 106 ccRCC patients were retrospectively analyzed. The relationship between RUNX3 methylation level and overall survivals was evaluated using the Kaplan-Meyer analysis and Cox regression analysis (univariate and multivariate). The relationship between RUNX3 methylation level and CT features was evaluated using chi-square test and logistic regression analysis (univariate and multivariate). RESULTS: ß value cutoff of 0.53 to distinguish high methylation (N = 44) from low methylation tumors (N = 62). Patients with lower levels of methylation had longer median overall survival (49.3 vs. 28.4) months (low vs. high, adjusted hazard ratio [HR] 4.933, 95% CI 2.054-11.852, p < 0.001). On univariate logistic regression analysis, four risk factors (margin, side, long diameter, and intratumoral vascularity) were associated with RUNX3 methylation level (all p < 0.05). Multivariate logistic regression analysis found that three risk factors (side: left vs. right, odds ratio [OR] 2.696; p = 0.024; 95% CI 1.138-6.386; margin: ill-defined vs. well-defined, OR 2.685; p = 0.038; 95% CI 1.057-6.820; and intratumoral vascularity: yes vs. no, OR 3.286; p = 0.008; 95% CI 1.367-7.898) were significant independent predictors of high methylation tumors. This model had an area under the receiver operating characteristic curve (AUC) of 0.725 (95% CI 0.623-0.827). CONCLUSIONS: Higher levels of RUNX3 methylation are associated with shorter survival in ccRCC patients. And presence of intratumoral vascularity, ill-defined margin, and left side tumor were significant independent predictors of high methylation level of RUNX3 gene. KEY POINTS: • RUNX3 methylation level is negatively associated with overall survival in ccRCC patients. • Presence of intratumoral vascularity, ill-defined margin, and left side tumor were significant independent predictors of high methylation level of RUNX3 gene.