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Previous studies have demonstrated that glucocorticoid receptor β (GRβ) functions as an oncoprotein, regulating the malignant phenotypes and stem-like cell maintaining in human glioblastoma (GBM). Of the glucocorticoid receptor (GR) isoforms, GRβ and GRα are highly homologous, though the mechanism underlying the distinct functions of these two isoforms in GBM has not been clarified. Here by establishing a carboxyl-terminal (COOH-terminal) deletion mutant, we determined that GRβ can be ubiquitinated. We also found that its COOH terminal is essential for this ubiquitination. The mutation of a lysine to arginine at residue 733 (K733R) blocked the ubiquitination of GRβ, indicating that K733 is a key site for ubiquitination. Using K733R to establish nonubiquitinated GRβ, we demonstrated that ubiquitination not only regulates the stability and nuclear translocation of GRβ, but is also a vital mechanism for its oncogenic functions in vitro and in vivo. Protein interaction assay further indicated that ubiquitin-specific protease 49 (USP49) is a GRβ-binding protein and the interaction depends on GRβ ubiquitination. USP49 knockdown resulted in a decrease of cell proliferation, invasion, and an increase of cell apoptosis. More importantly, USP49 knockdown increased ubiquitination and amplified the oncogenic effects of GRβ, confirming the decisive role of ubiquitination on GRβ carcinogenicity. Taken together, these findings established that ubiquitination is a vial process for GRβ the execution of oncogenic functions in GBM and that the K733 site is crucial for ubiquitination of GRβ. Implications: This work is the first identify of the activation GRβ by a single lysine point-mediated ubiquitination and proteasome degradation, which determines its oncogenic functions in GBM.

Tetrandrine (TET) is a potent calcium channel blocker used to treat hypertension and inflammation. Currently, TET is predominantly used to treat a variety of human diseases, and there is little information regarding the use of TET against plant pathogens. In this study, we explored the antifungal activity of TET on a plant pathogen, Botrytis cinerea. We show that administration of low concentrations of TET effectively inhibited hyphal growth of fungus grown on potato dextrose agarose and decreased the virulence of B. cinerea in tomato plants. Real-time PCR revealed that the expression of drug efflux pump-related genes (alcohol dehydrogenase 1, multidrug/pheromone exporter, pleiotropic drug resistance protein 1, and synaptic vesicle transporter) were downregulated in the presence of TET. Finally, we show that TET acts synergistically with iprodione, resulting in increased inhibition of B. cinerea both in vitro and in vivo. These results indicate that TET might act as an effective antifungal agent in reducing gray mold disease.

A secondary metabolites investigation on Biscogniauxia sp. 71-10-1-1 was carried out, which led to the obtention of nine new diisoprenyl-cyclohexene/ane-type meroterpenoids (1-9) and two new isoprenylbenzoic acid-type meroterpeniods (10-11). The structures of these isolates were established on the basis of multispectroscopic analyses, ECD, and 13C chemical shifts calculations, and single-crystal X-ray diffraction. Among them, biscognin A (1) is the first diisoprenyl-cyclohexene-type meroterpenoid with a unique 2-isopropyl-6'-methyloctahydro-1'H-spiro[cyclopropane-1,2'-naphthalene] skeleton. Biscognienyne F (5) is the first diisoprenyl-cyclohexene-type meroterpenoid with a cyclic carbonate. The anti-inflammatory assays of the majority of compounds were evaluated, which exhibited that compounds 3 and 5 can obviously inhibit pro-inflammatory cytokines TNF-α and IL-6 productions. This is the first report for diisoprenyl-cyclohexene-type meroterpenoids with anti-inflammatory activity. Moreover, the possible biogenetic pathways of the majority of compounds (1-5) are proposed.

Squamous cell carcinoma (SqCC) is the most common malignancy of the anal canal, where it is strongly associated with HPV infection. Characteristic genomic alterations have been identified in anal SqCC, but their clinical significance and correlation with HPV status, pathologic features, and immunohistochemical markers are not well established. We examined the molecular and clinicopathologic features of 96 HPV-positive and 20 HPV-negative anal SqCC. HPV types included 89 with HPV16, 2 combined HPV16/HPV18, and 5 HPV33. HPV-positive cases demonstrated frequent mutations or amplifications in PIK3CA (30%; p = 0.027) or FBXW7 mutations (10%). HPV-negativity was associated with frequent TP53 (53%; p = 0.00001) and CDKN2A (21%; p = 0.0045) mutations. P16 immunohistochemistry was positive in all HPV-positive cases and 3/20 HPV-negative cases (p < 0.0001; sensitivity: 100%; specificity: 85%) and was associated with basaloid morphology (p = 0.0031). Aberrant p53 immunohistochemical staining was 100% sensitive and specific for TP53 mutation (p < 0.0001). By the Kaplan-Meier method, HPV-negativity, aberrant p53 staining, and TP53 mutation were associated with inferior overall survival (OS) (p < 0.0001, p = 0.0103, p = 0.0103, respectively) and inferior recurrence-free survival (p = 0.133, p = 0.0064, and p = 0.0064, respectively). TP53/p53 status stratified survival probability by HPV status (p = 0.013), with HPV-negative/aberrant p53 staining associated with the worst OS, HPV-positive/wild-type p53 with best OS, and HPV-positive/aberrant p53 or HPV-negative/wild-type p53 with intermediate OS. On multivariate analysis HPV status (p = 0.0063), patient age (p = 0.0054), T stage (p = 0.039), and lymph node involvement (p = 0.044) were independently associated with OS. PD-L1 expression (CPS ≥ 1) was seen in 30% of HPV-positive and 40% of HPV-negative cases, and PD-L1 positivity was associated with a trend toward inferior OS within the HPV-negative group (p = 0.064). Our findings suggest that anal SqCC can be subclassified into clinically, pathologically, and molecularly distinct groups based on HPV and TP53 mutation status, and p16 and p53 immunohistochemistry represent a clinically useful method of predicting these prognostic groups.

Bacteria infection and laryngopharyngeal reflux (LPR) were believed the important pathogenesis of chronic otitis media with effusion (COME). But no study researched the relationship between them on COME.To confirm bacterial could arrive middle ear through LPR and produced acid metabolites to activate the pepsinogen of LPR causing COME.Children (65) diagnosed COME with 122 middle ear effusions were included in COME group. Children (22) with congenital/acquired profound deafness with 22 middle ear lavage were included in CI group. Pepsin A concentration in the effusion and lavage fluid were measured. The DNA of the bacteria, IL-8 and TNF-α in the effusion were detected.The average concentration of pepsin A in the effusions and lavage were 176.65 ± 242.09 and 19 ng/ml. Bacterial infection rates were 75.76% and 24.24% in the pepsin A(+) and pepsin A(-) patients. In the bacterial (+), the patients of pepsin A(+) was 4.33 times higher than those of pepsin A(-). TNF-α in pepsin A(+) was higher than that in pepsin A(-). TNF-α and IL-8 were higher in bacteria(+) than those of bacteria(-).Bacterial infection and LPR might act in synergy in the pathogenesis of COME.First time to propose LPR and bacterial infection might work synergistically to cause COME.

Cavernous malformations affecting the CNS occur in ∼0.16–0.4% of the general population. The majority (85%) of cavernous malformations are in a sporadic form, but the genetic background of sporadic cavernous malformations remains enigmatic. Of the 81 patients, 73 (90.1%) patients were detected carrying somatic missense variants in two genes: MAP3K3 and PIK3CA by whole-exome sequencing. The mutation spectrum correlated with lesion size (P = 0.001), anatomical distribution (P

Background Tracheobronchial foreign body aspiration (TFBA) is a critical disease in children and is extremely dangerous, even life-threatening. The factors affecting the occurrence and prognosis of TFBA are complex. The purpose of this study is to examine the external and intrinsic factors affecting clinical features of TFBA in West China and propose potential effective intervention measures. Methods We retrospectively analyzed the clinical data of pediatric patients diagnosed with TFBA with foreign bodies (FBs) removed by rigid bronchoscopy under general anesthesia at the otolaryngology department from December 2017 to November 2018. The data included age, sex, clinical symptoms, type and location of FB, guardians, prehospital duration and residence of these pediatric patients. Results The ratio of males (72) to females (53) was 1.4:1. Children aged from 1 to 3 years accounted for 76% (95/125) of patients. Cough, continuous fever and dyspnea were the primary symptoms. The right primary bronchus was the most common location of FB detection by rigid bronchoscopy (67 cases, 53.6%). Organic FBs were most common in our study. Guardians of patients significantly differed in the rural (parents 16, grandparents 31) and urban (parents 52, grandparents 26) groups (χ2 = 12.583, p = 0.000). More children in the rural group than in the urban group had a treatment delay longer than 72 h. More children in the group with no history of FB aspiration (12, 25%) than in the group with prior FB aspiration had a treatment delay longer than 72 h. Conclusion Pediatric TFBA is a common emergency in otolaryngology. Age, sex, tracheobronchial anatomy and other physiological elements were defined as intrinsic factors, while guardians, residence, FB species and prehospital time were defined as external factors of TFBA. External and intrinsic factors both influence the occurrence and progression of TFBA. It is extremely important to take effective measures to control external factors, which can decrease morbidity and mortality.

Introduction Chronic rhinosinusitis is a highly heterogeneous chronic inflammation of the upper respiratory tract caused by immune dysfunction in human beings. However, the underlying etiology of this disease has not yet been well established. Several trials have revealed that serum vitamin D level abnormality might play a role in the pathophysiology of chronic rhinosinusitis. Objective The aim of present study was to perform a meta-analysis of studies to compare the serum vitamin D levels between patients with chronic rhinosinusitis and healthy controls and to evaluate the potential associations of serum vitamin D level with the occurrence of chronic rhinosinusitis. Methods Following the PRISMA guidelines, relevant databases incorporating Pubmed, Web of Science, Embase and Cochrane Library were searched from inception to December 1, 2018. Funnel plot symmetry and Egger’ test were used to detect publication biases. The outcomes were presented as Weighted Mean Difference combined with 95% Confidence Intervals to estimate the difference of serum vitamin D levels between patients with chronic rhinosinusitis and controls. Higgins I2 value was used to test for heterogeneity between the included studies. Results Out of 176 studies, only eight studies involving 337 chronic rhinosinusitis patients and 179 healthy controls met the criteria and were included in the meta-analysis. In a pooled analysis of the studies, chronic rhinosinusitis patients showed decreased serum vitamin D status (WMD = −7.80, 95% CI −13.28 to −2.31, p = 0.000). Subgroups analyses based on study location (USA vs. Non-USA), types of biomarkers (25(OH)D3 vs. 25(OH)D) and the study design methods (retrospective vs. prospective) did not reveal heterogeneity. However, phenotypes of chronic rhinosinusitis, with versus without polyposis might account for some degree of heterogeneity. Meanwhile, a lower serum vitamin D level was observed in chronic rhinosinusitis with nasal polyps patients. Conclusion Our findings indicate that serum vitamin D level might be associated with patients with chronic rhinosinusitis as we detected a significant association between lower serum vitamin D status and chronic rhinosinusitis, especially in chronic rhinosinusitis with nasal polyps patients. However, further comprehensive studies are warranted in order to draw firm conclusions.

Tian Shen,1,* Jing Wang,1,* Wen Yang,1 Linke Li,1 Yixin Qiao,1 Xiaohong Yan,1 Min Chen,2 Xiangdong Tang,3 Jian Zou,1 Yu Zhao1 1Department of Otorhinolaryngology Head and Neck Surgery, West China Hospital, Sichuan University, Chengdu, Sichuan, People’s Republic of China; 2Department of Otorhinolaryngology Head and Neck Surgery, Chengdu Shangjin Nanfu Hospital, West China Hospital, Sichuan University, Chengdu, Sichuan, People’s Republic of China; 3Sleep Medicine Center, West China Hospital, Sichuan University, Chengdu, Sichuan, People’s Republic of China*These authors contributed equally to this workCorrespondence: Yu Zhao; Jian ZouDepartment of Otolaryngology Head and Neck Surgery, West China Hospital, Sichuan University, Chengdu, Sichuan, People’s Republic of ChinaTel +86-18980601410; +86-18980605883Email yutzhao@163.com; zoujian926@163.comObjective: To investigate the possible correlation between obesity and hematological parameters in children with obstructive sleep apnea (OSA).Methods: A total of 460 children was initially included in our study, which were divided into children with OSA and children without OSA. Multiple analysis was performed for obesity by adjusting confounding factors such as age and gender in 460 children. Further, to minimize the influence of confounding factors and selective bias, propensity score matching (PSM) was performed in children with OSA. Hematological parameters such as inflammatory and coagulable parameters were compared between the normal weight children with OSA and the obese children with OSA following PSM.Results: OSA (OR = 3.061; P< 0.001; 95% CI, 1.772– 5.288) represented an independent risk factor for obesity. Besides, the obese children with OSA had higher levels of white blood cell (WBC) (P< 0.001), neutrophil (NEUT) (P< 0.001), neutrophil-lymphocyte ratio (NLR) (P=0.006), fibrinogen (FIB) (P=0.033), while had a lower level of activated partial thromboplastin time (APTT) (P=0.048). No significant differences were observed in other hematological parameters. In linear regression, the results indicated that the levels of WBC (R2 = 0.123, Beta = 0.289, P< 0.001), NEUT (R2 = 0.124, Beta = 0.282, P< 0.001), NLR (R2 = 0.105, Beta = 0.184, P=0.026) and FIB (R2 = 0.086, Beta = 0.246, P=0.003) were positively correlated with BMI, while the level of APTT (R2 = 0.057, Beta = − 0.171, P=0.044) was significantly negatively correlated with BMI.Conclusion: OSA was an independent risk factor contributing to obesity. WBC, NEUT, NLR, FIB and APTT are correlated with obesity in children with OSA (aged from 2 to 14 years). These indicators could be used to estimate the status of inflammation and hypercoagulation in the obese children with OSA.Keywords: obstructive sleep apnea, obesity, propensity score matching, hematological parameter, inflammation, hypercoagulation

Alcohol use disorder (AUD) pathology features pro‐inflammatory gene induction and microglial activation. The underlying cellular processes that promote this activation remain unclear. Previously considered cellular debris, extracellular vesicles (EVs) have emerged as mediators of inflammatory signaling in several disease states. We investigated the role of microvesicles (MVs, 50 nm–100 µm diameter EVs) in pro‐inflammatory and microglial functional gene expression using primary organotypic brain slice culture (OBSC). Ethanol caused a unique immune gene signature that featured: temporal induction of pro‐inflammatory TNF‐α and IL‐1β, reduction of homeostatic microglia state gene Tmem119, progressive increases in purinergic receptor P2RY12 and the microglial inhibitory fractalkine receptor CX3CR1, an increase in the microglial presynaptic gene C1q, and a reduction in the phagocytic gene TREM2. MV signaling was implicated in this response as reduction of MV secretion by imipramine blocked pro‐inflammatory TNF‐α and IL‐1β induction by ethanol, and ethanol‐conditioned MVs (EtOH‐MVs) reproduced the ethanol‐associated immune gene signature in naïve OBSC slices. Depletion of microglia prior to ethanol treatment prevented pro‐inflammatory activity of EtOH‐MVs, as did incubation of EtOH‐MVs with the HMGB1 inhibitor glycyrrhizin. Ethanol caused HMGB1 secretion from cultured BV2 microglia in MVs through activation of PI3 kinase. In summary, these studies find MVs modulate pro‐inflammatory gene induction and microglial activation changes associated with ethanol. Thus, MVs may represent a novel therapeutic target to reduce neuroinflammation in the setting of alcohol abuse or other diseases that feature a neuroimmune component. [Correction added on 5 April 2021, after first online publication: The copyright line was changed.], Ethanol causes the secretion of pro‐inflammatory microvesicles (MVs) from brain tissue. Ethanol‐induced MVs reproduce the pro‐inflammatory gene induction and microglial activation seen with ethanol alone, and inhibition of their secretion blocks ethanol‐induced neuroinflammation. Microglia are required for the generation of pro‐inflammatory MVs, with MV secretion involving activation of microglial PI3K.

Macroautophagy/autophagy is known to be important for intracellular quality control in the lens. GJA8 is a major gap junction protein in vertebrate lenses. Mutations in GJA8 cause cataracts in humans. The well-known cataractogenesis mechanism is that mutated GJA8 leads to abnormal assembly of gap junctions, resulting in defects in intercellular communication among lens cells. In this study, we observed that ablation of Gja8b (a homolog of mammalian GJA8) in zebrafish led to severe defects in organelle degradation, an important cause of cataractogenesis in developing lens. The role of autophagy in organelle degradation in lens remains disputable. Intriguingly, we also observed that ablation of Gja8b induced deficient autophagy in the lens. More importantly, in vivo treatment of zebrafish with rapamycin, an autophagy activator that inhibits MAPK/JNK and MTORC1 signaling, stimulated autophagy in the lens and relieved the defects in organelle degradation, resulting in the mitigation of cataracts in gja8b mutant zebrafish. Conversely, inhibition of autophagy by treatment with the chemical reagent 3-MA blocked these recovery effects, suggesting the important roles of autophagy in organelle degradation in the lens in gja8b mutant zebrafish. Further studies in HLE cells revealed that GJA8 interacted with ATG proteins. Overexpression of GJA8 stimulated autophagy in HLE cells. These data suggest an unrecognized cataractogenesis mechanism caused by ablation of Gja8b and a potential treatment for cataracts by stimulating autophagy in the lens. Abbreviations: 3-MA: 3-methyladenine; ATG: autophagy related; AV: autophagic vacuoles; Dpf: days post fertilization; GJA1: gap junction protein alpha 1; GJA3: gap junction protein alpha 3; GJA8: gap junction protein alpha 8; Hpf: hours post fertilization; MAP1LC3/LC3: microtubule associated protein 1 light chain 3; MTOR: mechanistic target of rapamycin kinase; PtdIns3K: class III phosphatidylinositol 3-kinase; WT: wild type.

Circular RNAs (circRNAs) function as efficient microRNA (miRNA) sponges that regulate gene expression in the pathogenesis of many human malignancies. However, their roles in cervical adenocarcinoma remain largely unknown. In this study, we aimed to seek novel circRNAs that regulate cervical adenocarcinoma carcinogenesis and to explore their regulatory mechanisms as well as clinical significance. We identified that 24 circRNAs were differentially expressed in cervical adenocarcinoma tissues by RNA sequencing. Among them, circEYA1 was the most significantly downregulated circRNA in cervical adenocarcinoma. In cervical adenocarcinoma cells, circEYA1 overexpression led to suppression of cell viability and colony formation, promotion of apoptosis, and a decrease of the xenograft tumor growth. The mechanism underlying these observations is that circEYA1 functioned as a sponge of miR-582-3p and abrogated its suppression of CXCL14 expression. Consistently, miR-582-3p inhibition phenocopied the biological effects of circEYA1 overexpression in cervical adenocarcinoma cells. Moreover, miR-582-3p overexpression reversed the suppressive behaviors of circEYA1 in vitro and in vivo. In addition, the expression, correlation, and clinical diagnostic value of circEYA1/miR-582-3p/CXCL14 were confirmed in 198 clinical cervical tissue samples. In summary, our findings highlight a novel tumor suppressive role of circEYA1 in cervical adenocarcinoma tumorigenesis and may provide a potential diagnostic marker and therapeutic target for patients with cervical adenocarcinoma., Graphical Abstract, Xu et al. explored the expression profile of circRNAs in cervical adenocarcinoma. circEYA1 functions as a sponge of miR-582-3p to abolish the suppressive effect of miR-582-3p on CXCL14 and then inhibits cell viability. The findings may provide a potential diagnostic marker and therapeutic target for patients with cervical adenocarcinoma.

Despite the fact that the incidence of cervical squamous cell carcinoma has decreased, there is an increase in the incidence of cervical adenocarcinoma. However, our knowledge on cervical adenocarcinoma is largely unclear. Transcriptome sequencing was conducted to compare 4 cervical adenocarcinoma tissue samples with 4 normal cervical tissue samples. mRNA, lncRNA, and miRNA signatures were identified to discriminate cervical adenocarcinoma from normal cervix. The expression of VIL1, HNF1A-AS1, MIR194-2HG, SSTR5-AS1, miR-192-5p, and miR-194-5p in adenocarcinoma were statistically significantly higher than that in normal control samples. The Receiver Operating Characteristic (ROC) curve analysis indicated that combination of miR-192-5p, HNF1A-AS1, and VIL1 yielded a better performance (AUC = 0.911) than any single molecule -and could serve as potential biomarkers for cervical adenocarcinoma. Of note, the combination model also gave better performance than TCT test for cervical adenocarcinoma diagnosis. However, there was no correlation between miR-192-5p or HNF1A-AS1 and HPV16/18 E6 or E7. VIL1 was weakly correlated with HPV18 E7 expression. In summary, our study has identified miR-192-5p/HNF1A-AS1/VIL1 panel that accurately discriminates adenocarcinoma from normal cervix. Detection of this panel may provide considerable clinical value in the diagnosis of cervical adenocarcinoma.

Background Images from magnetic resonance imaging (MRI) are crucial unstructured data for prognostic evaluation in nasopharyngeal carcinoma (NPC). We developed and validated a prognostic system based on the MRI features and clinical data of locoregionally advanced NPC (LA-NPC) patients to distinguish low-risk patients with LA-NPC for whom concurrent chemoradiotherapy (CCRT) is sufficient. Methods This multicenter, retrospective study included 3444 patients with LA-NPC from January 1, 2010, to January 31, 2017. A 3-dimensional convolutional neural network was used to learn the image features from pretreatment MRI images. An eXtreme Gradient Boosting model was trained with the MRI features and clinical data to assign an overall score to each patient. Comprehensive evaluations were implemented to assess the performance of the predictive system. We applied the overall score to distinguish high-risk patients from low-risk patients. The clinical benefit of induction chemotherapy (IC) was analyzed in each risk group by survival curves. Results We constructed a prognostic system displaying a concordance index of 0.776 (95% confidence interval [CI] = 0.746 to 0.806) for the internal validation cohort and 0.757 (95% CI = 0.695 to 0.819), 0.719 (95% CI = 0.650 to 0.789), and 0.746 (95% CI = 0.699 to 0.793) for the 3 external validation cohorts, which presented a statistically significant improvement compared with the conventional TNM staging system. In the high-risk group, patients who received induction chemotherapy plus CCRT had better outcomes than patients who received CCRT alone, whereas there was no statistically significant difference in the low-risk group. Conclusions The proposed framework can capture more complex and heterogeneous information to predict the prognosis of patients with LA-NPC and potentially contribute to clinical decision making.

A number of studies have reported that diabetic retinopathy (DR) is the major cause of blindness. Berberine (BBR) is a bioactive constituent that displays effects on blood glucose; however, the mechanism underlying the role of BBR during the development of DR is not completely understood. In the present study, a rat model of DR was successfully established. The eye tissues were removed and subsequently assessed by hematoxylin and eosin staining and the TUNEL assay. The catalase, malondialdehyde, reactive oxygen species, glutathione and superoxide dismutase contents of the eye tissues were measured. Muller cells were chosen for further in vitro experiments. Cell apoptosis was examined by Annexin V‑FITC apoptosis detection and Hoechst staining, and the mitochondrial membrane potential was assessed by JC‑1 mitochondrial membrane potential detection. BBR decreased ganglion cell layer, cell apoptosis, reduced diabetic‑induced oxidative stress and deactivated the NF‑κB signaling pathway in the rat model of DR. High glucose enhanced oxidative stress and induced mitochondria‑dependent cell apoptosis in Muller cells by activating the NF‑κB signaling pathway. BBR reversed the high glucose‑induced effects by decreasing the phosphorylation of IκB, inhibiting NF‑κB nuclear translocation and deactivating the NF‑κB signaling pathway. The results suggested that BBR protected against DR by inhibiting oxidative stress and cell apoptosis via deactivation of the NF‑κB signaling pathway; therefore, suggesting that BBR may serve as a promising therapeutic agent for DR.

Background and purposeNuclear grades of clear cell renal cell carcinoma (ccRCC) are usually confirmed by invasive methods. Radiomics is a quantitative tool that uses non-invasive medical imaging for tumor diagnosis and prognosis. In this study, a radiomics approach was proposed to analyze the association between preoperative computed tomography (CT) images and nuclear grades of ccRCC.MethodsOur dataset included 320 ccRCC patients from two centers and was divided into a training set (n = 124), an internal test set (n = 123), and an external test set (n = 73). A radiomic feature set was extracted from unenhanced, corticomedullary phase, and nephrographic phase CT images. The maximizing independent classification information criteria function and recursive feature elimination with cross-validation were used to select effective features. Random forests were used to build a final model for predicting nuclear grades, and area under the receiver operating characteristic curve (AUC) was used to evaluate the performance of radiomic features and models.ResultsThe radiomic features from the three CT phases could effectively distinguished the four nuclear grades. A combined model, merging radiomic features and clinical characteristics, obtained good predictive performances in the internal test set (AUC 0.77, 0.75, 0.79, and 0.85 for the four grades, respectively), and performance was further confirmed in the external test set, with AUCs of 0.75, 0.68, and 0.73 (no fourth-level data).ConclusionThe combination of CT radiomic features and clinical characteristics could discriminate the nuclear grades in ccRCC, which may help in assisting treatment decision making.

Background Laparoscopic Roux-en-Y gastric bypass is a leading metabolic surgery for obese patients with type 2 diabetes mellitus. However, postoperative nutritional deficiencies and abnormal bone metabolism are possible. In this study, we investigated the mid-term impact on bone metabolism of laparoscopic Roux-en-Y gastric bypass. Methods This retrospective study involved 238 patients with type-2 diabetes mellitus and obesity who underwent laparoscopic Roux-en-Y gastric bypass in our institution; patients were followed for 3 years. Patients received a calcium supplement (1,200 mg per day), alfacalcidol (0.5 µg per day), and a multivitamin (1 tablet per day), postoperatively. We recorded and analyzed patients’ body mass index, and blood levels of 25-hydroxy-vitamin D, calcium, and parathyroid hormone. Data for 16 patients 6 months postoperatively were selected randomly to evaluate bone-specific alkaline phosphatase and the bone resorption marker, C-terminal crosslinking telopeptide of type I collagen (CTX-1). Results Of the 238 patients, 111 were women, and 127 were men (mean age: 46.91±12.03 years; mean body mass index: 31.37 kg/m2). 25-hydroxy vitamin D3 levels did not change (P>0.05) compared with preoperative levels during the 3-year follow-up. Parathyroid hormone (PTH) levels increased (P0.05). However, CTX-1 levels increased in the first 12 months postoperatively, then decreased in the second year; bone alkaline phosphatase levels increased gradually. Conclusions Laparoscopic Roux-en-Y gastric bypass is effective for weight loss, but also has an impact on bone metabolism, which increases the risk of fracture. Our results highlight the importance of regular follow-ups and full-dose calcium and vitamin supplementation, postoperatively.

Supplemental Digital Content is available in the text, Background Hyperuricemia and gout have become public health concerns; many important guidelines have recommended xanthine oxidase inhibitors (XOIs) as the first-line urate-lowering therapies (ULTs) to treat chronic gout with hyperuricemia. However, whether treating hyperuricemia and gout with ULTs modifies cardiovascular risks remains controversial. The aim of this study was to assess the incident risk of cardiovascular (CV) events (CVE) in hyperuricemia population, assess the cardiovascular benefit-risk of ULTs in hyperuricemia patients with or without gout in diverse cardiovascular risk sub-groups, and specify the safety of different ULTs. Methods We searched PubMed, Embase, the Cochrane Library, Wanfang, Chongqing VIP (CQVIP, en.cqvip.com), and China National Knowledge Infrastructure Database for prospective cohort studies and randomized controlled trials (RCTs) in English and Chinese. Potential medications included XOIs, and uricosurics. RCTs were divided into sub-groups analysis based on blinding status and patients’ history of CV diseases. Risk ratios (RRs) were calculated and were reported with corresponding 95% confidence intervals (CIs) by fixed-effects or random-effects model. Results Seven prospective cohort studies and 17 RCT studies were included. The risks of both major adverse cardiovascular events (MACE) (RR = 1.72, 95% CI 1.28–2.33) and CVE (RR = 1.35, 95% CI 1.12–1.62) were higher in the hyperuricemia population than non-hyperuricemia one. In seven RCT studies where XOIs were compared with no-treatment or placebo, the results of five low CV risk studies showed that XOIs lowered the risks of both MACE (RR = 0.35, 95% CI 0.20–0.62) and CVE (RR = 0.61, 95% CI 0.44–0.85); whereas two high CV risk studies showed that XOIs lowered the risk of CVE (RR = 0.69, 95% CI 0.54–0.88) rather than MACE (RR = 0.62, 95% CI 0.29–1.35). In nine RCT studies where the cardiovascular safety between febuxostat and allopurinol were compared, no statistical difference was found in the risk of MACE or CVE. Conclusions The hyperuricemia population does have a higher incidence of CVE, and the results suggested that XOIs might reduce the incidence of MACE and total CVE. In addition, from the perspective of cardiovascular safety, febuxostat equaled allopurinol in our meta-analysis.

BACKGROUND Gleason score (GS) is a histologic prognostic factor and the basis of treatment decision-making for prostate cancer (PCa). Treatment regimens between lower-grade (GS ≤7) and high-grade (GS >7) PCa differ largely and have great effects on cancer progression. PURPOSE To investigate the use of different sequences in biparametric MRI (bpMRI) of the prostate gland for noninvasively distinguishing high-grade PCa. STUDY TYPE Retrospective. POPULATION In all, 489 patients (training cohort: N = 326; test cohort: N = 163) with PCa between June 2008 and January 2018. FIELD STRENGTH/SEQUENCE 3.0T, pelvic phased-array coils, bpMRI including T2 -weighted imaging (T2 WI) and diffusion-weighted imaging (DWI); apparent diffusion coefficient map extracted from DWI. ASSESSMENT The whole prostate gland was delineated. Radiomic features were extracted and selected using the Kruskal-Wallis test, the minimum redundancy-maximum relevance, and the sequential backward elimination algorithm. Two single-sequence radiomic (T2 WI, DWI) and two combined (T2 WI-DWI, T2 WI-DWI-Clinic) models were respectively constructed and validated via logistic regression. STATISTICAL TESTS The Kruskal-Wallis test and chi-squared test were utilized to evaluate the differences among variable groups. P

Age-related macular degeneration (AMD), a progressive chronic disease of the central retina, is a leading cause of blindness worldwide. Activated macrophages recruited to the injured eyes greatly contribute to the pathogenesis of choroidal neovascularization (CNV) in exudative AMD (wet AMD). This study describes the effects of cyclooxygenase-2 (COX2)/prostaglandin E2 (PGE2) signalling on the M2 macrophage recruitment and CNV formation of wet AMD. In a mouse model of laser-induced wet AMD, the mice received an intravitreal injection of celecoxib (a selective COX2 inhibitor). Optical coherence tomography (OCT), fundus fluorescein angiography (FFA), choroidal histology of the CNV lesions, and biochemical markers were assessed. The level of PGE2 expression was high in the laser-induced CNV lesions. M2 polarization and CNV development were significantly less after celecoxib treatment. E-prostanoid1 receptor (EP1R)/protein kinase C (PKC) signalling was involved in M2 polarization and interleukin-10 (IL-10) production of bone marrow-derived macrophages (BMDMs) in vitro. In addition, IL-10 was found to induce the proliferation and migration of human choroidal microvascular endothelial cells (HCECs). Thus, the PGE2/EP1R signalling network serves as a potential therapeutic target for CNV of the wet-type AMD.

Gastrointestinal stromal tumors (GISTs) are the most common mesenchymal tumors of the gastrointestinal tract. As the main GIST drivers, gain-of-function mutations inKITorPDGFRAare closely associated with not only tumor development and progression but also therapeutic response. In addition to the status of KIT and PDGFRA, little is known about other potential GIST-related genes. In this study, we identified the mutation profiles in 49 KIT-mutated GIST tumors using the whole exome sequencing (WES) method. Furthermore, some representative mutations were further validated in an independent GIST cohort using the SNaPshot SNP assay. We identified extensive and diverse mutations of KIT in GIST, including many undescribed variants. In addition, we revealed some new tumor-related gene mutations with unknown pathogenicity. By enrichment analyses of gene function and protein-protein interaction network construction, we showed that these genes were enriched in several important cancer- or metabolism-related signaling pathways, including PI3K-AKT,RTK-RAS, Notch, Wnt, Hippo, mTOR, AMPK, and insulin signaling. In particular, DNA repair-related genes, includingMLH1,MSH6,BRCA1,BRCA2, andPOLE, are frequently mutated in GISTs, suggesting that immune checkpoint blockade may have promising clinical applications for these GIST subpopulations. In conclusion, in addition to extensive and diverse mutations ofKIT, some genes related to DNA-repair and cell metabolism may play important roles in the development, progression and therapeutic response of GIST.

Description, HSPA13 is a novel checkpoint regulator in the TNFα signaling pathway controlling cellular physiological outcomes., RIP1 has emerged as a master regulator in TNFα signaling that controls two distinct cellular fates: cell survival versus programmed cell death. Because the default response of most cells to TNFα is NF-κB–mediated inflammation and survival, a specific mechanism must exist to control the divergence of signaling outcome. Here, we identify HSPA13 as a transcription-independent checkpoint to modulate the role of RIP1 in TNFα signaling. Through specific binding to TNFR1 and RIP1, HSPA13 enhances TNFα-induced recruitment of RIP1 to TNFR1, and consequently promotes downstream NF-κB transcriptional responses. Meanwhile, HSPA13 attenuates the participation of RIP1 in cytosolic complex II and prevents cells from programmed death. Loss of HSPA13 shifts the transition of RIP1 from complex I to complex II and promotes both apoptosis and necroptosis. Thus, our study provides compelling evidence for the cellular protective function of HSPA13 in fine-tuning TNFα responses.

Retinal pigment epithelium (RPE) cell damage is implicated in the pathogenesis of age-related macular degeneration (AMD). An increase of interferon-γ (IFN-γ) levels was observed in patients with AMD, but whether inflammatory factors are causally related to AMD progression is unclear. Here, we demonstrate a direct causal relationship between IFN-γ and RPE cell death. IFN-γ induced human retinal pigment epithelial cell (ARPE-19) death accompanied by increases in Fe2+ , reactive oxygen species, lipid peroxidation, and glutathione (GSH) depletion, which are main characteristics of ferroptosis. Mechanistically, IFN-γ upregulates the level of intracellular Fe2+ through inhibiting Fe2+ efflux protein SLC40A1 and induces GSH depletion by blocking cystine/glutamate antiporter, System xc-. At the same time, treatment with IFN-γ decreases the level of glutathione peroxidase 4 (GPx4), rendering the cells more sensitive to ferroptosis. JAK1/2 and STAT1 inhibitors could reverse the reduction of SLC7A11, GPx4 and GSH expression induced by IFN-γ, indicating IFN-γ induces ARPE-19 cell ferroptosis via activation of the JAK1-2/STAT1/SLC7A11 signaling pathway. The above results were largely confirmed in IFN-γ-treated mice in vivo. Finally, we used sodium iodate (NaIO3 )-induced retinal degeneration to further explore the role of ferroptosis in AMD in vivo. Consistent with the role of IFN-γ, treatment with NaIO3 decreased SLC7A11, GPx4 and SLC40A1 expressions. NaIO3 -induced RPE damage was accompanied by increased iron, lipid peroxidation products (4-hydroxynonenal, malondialdehyde), and GSH depletion, and ferroptosis inhibitors could reverse the above phenomenon. Taken together, our findings suggest that inhibiting ferroptosis or reducing IFN-γ may serve as a promising target for AMD.

Neutrophils are rapidly recruited from the peripheral blood to the inflammatory site to initiate inflammatory response against pathogenic infections. The process to recruit neutrophils must be properly regulated since the abnormal accumulation of neutrophils can cause organ damage and dysfunction. The acute respiratory distress syndrome (ARDS)/acute lung injury (ALI) is a common cause of respiratory failure that is characterized by the infiltration of neutrophils and epithelial integrity disruption. Indeed, recent studies suggest a pathogenic role of neutrophils in the clinic severity of the coronavirus disease 2019 (COVID-19) ARDS. The chemokine CXCL1, which is rapidly induced by inflammatory stimuli, plays a key role in neutrophil influx during lung inflammation. The molecular basis ofCxcl1induction is not fully understood. Here we report that TET1, a member of the ten eleven translocation (TET) methylcytosine dioxygenase protein family, displays a striking specificity in the regulation of gene expression in macrophages. RNA sequencing (RNA-seq) analysis showed thatTet1disruption significantly altered the expression of only 48 genes that includeCxcl1and several other genes known to be important for cell migration and trafficking in bone marrow derived macrophages (BMDMs) in response to LPS stimulation. TET1 regulates the induction ofCxcl1by facilitating the DNA demethylation of theCxcl1promoter. InTet1−/−mice, the induction ofCxcl1was suppressed, resulting in defective neutrophil recruitment to the lung during LPS-induced acute lung injury. Our results identify a novel epigenetic mechanism that selectively controlsCxcl1induction and neutrophil recruitment during acute lung injury.Key PointsTET1 has a striking specificity in macrophage gene regulation and controlsCxcl1induction by inflammatory stimuli via DNA demethylationNeutrophil recruitment is defective inTet1deficient mice during acute lung injury

The relationship between normal thyroid function and type 2 diabetes mellitus (T2DM) has been a particular focus for concern. The present study determined the relationship between thyroid hormone levels and the prevalence of diabetic retinopathy (DR) in T2DM patients. A cross-sectional study (n = 633) was performed in Xi’an, Shaanxi Province, China. Subjects were evaluated for anthropometric measurements, thyroid function, and diabetic retinopathy. Logistic regression models were used to assess the relationships between thyroid hormones and DR. Of 633 patients, 243 (38.4%) patients suffered from DR. The prevalence of DR showed a significantly decreasing trend across the quartiles based on free triiodothyronine (FT3) (FT3 quartile 1 group [FT3-Q1] P<0.001). In comparison with all participants categorized in FT3-Q1, the multivariable adjusted odds ratios (95% confidence interval) of DR in FT3-Q2, FT3-Q3, and FT3-Q4 were 0.587 (0.340–1.012), 0.458 (0.258–0.813), and 0.368 (0.201–0.673), (P=0.055, P=0.008, P=0.001), respectively. FT3 levels within the normal range are negatively associated with DR in euthyroid patients with type 2 diabetes. Further studies should be aimed at clarifying the relationship between thyroid hormones and T2DM.

Background: Diabetic retinopathy is a common microvascular complication of diabetes mellitus. Abnormal energy metabolism in microvascular endothelium is involved in the progression of diabetic retinopathy. Bile Acid G-Protein-Coupled Membrane Receptor (TGR5) has emerged as a novel regulator of metabolic disorders. However, the role of TGR5 in diabetes mellitus-induced microvascular dysfunction in retinas is largely unknown. Methods: ELISA was used for analyzing bile acid profiles in diabetic rat retinas and retinal microvascular endothelial cells (RMECs) cultured in high glucose medium. The effects of TGR5 agonist on streptozotocin-induced diabetic retinopathy were evaluated by HE staining, TUNEL staining, retinal trypsin digestion and vascular permeability assay. A pharmacological inhibitor of RhoA was used to study the role of TGR5 on the regulation of Rho/Rho-associated coiled-coil containing protein kinase (ROCK) and western blot, immunofluorescence and siRNA silencing were performed to study the related signaling pathways. Findings: A decrease in bile acids was observed during DR progression in the diabetic rat retinas and RMECs cultured in high glucose medium. The TGR5 agonist obviously ameliorated diabetes-induced retinal microvascular dysfunction in vivo, and inhibited the effect of TNF-α on endothelial cell proliferation, migration, and permeability in vitro. On the contrary, knockdown of TGR5 by siRNA aggravated TNF-α-induced actin polymerization and endothelial permeability. Mechanistically, the effects of TGR5 on the improvement of endothelial function was due to its regulatory role on the ROCK signaling pathway. An inhibitor of RhoA significantly reversed the loss of tight junction protein under TNF-α stimulation. Interpretation: Our findings suggest that insufficient bile acid signaling plays an important pathogenic role in the development of DR. Up-regulation or activation of TGR5 may inhibit RhoA/ROCK-dependent actin remodeling and represent an important therapeutic intervention for DR. Funding Statement: This project was supported by the Natural Science Foundation of China (81770941, 81800845), Nanjing Medical University Science and Technology Development Fund (NMUB2018363) and Natural Science Foundation of Jiangsu Province (BK20190149, BK20180170). Declaration of Interests: The authors declare that they have no competing interests. Ethics Approval Statement: All animal experiments were performed according to the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health, and the procedures were approved by the Animal Care and Use Committee of Nanjing Medical University.

BackgroundMicroglia are critical mediators of neuroimmune pathology across multiple neurologic disorders. Microglia can be persistently activated or “primed” by Toll-like receptor (TLR) activation, ethanol, stress, and other insults. Thus, strategies to prevent or reverse microglial priming may be beneficial for conditions that involve progressively increasing microglial activation. Microglial depletion with repopulation is emerging as a potential therapy to normalize chronic immune activation. Primary organotypic hippocampal slice culture (OHSC) allows for the study of neuroimmune activation as well as microglial depletion and repopulation without involvement of peripheral immune activation. OHSC undergoes functional maturation and retains cytoarchitecture similar toin vivo.MethodsOHSC underwent microglial depletion with the CSF1R antagonist PLX3397 with or without repopulation after removal of PLX3397. Immune, trophic, and synaptic gene changes in response to agonists of TLRs 2, 3, 4, 7, and 9 as well as ethanol were assessed in the settings of microglial depletion and repopulation. Gi-DREADD inhibition of microglia was used to confirm select findings seen with depletion. The ability of microglial repopulation to prevent progressive proinflammatory gene induction by chronic ethanol was also investigated.ResultsMicroglia were depleted (> 90%) by PLX3397 in OHSC. Microglial depletion blunted proinflammatory responses to several TLR agonists as well as ethanol, which was mimicked by Gi-DREADD inhibition of OHSC microglia. Removal of PLX3397 was followed by complete repopulation of microglia. OHSCs with repopulated microglia showed increased baseline expression of anti-inflammatory cytokines (e.g., IL-10), microglial inhibitory signals (e.g., CX3CL1), and growth factors (e.g., BDNF). This was associated with blunted induction (~ 50%) of TNFα and IL-1β in response to agonists to TLR4 and TLR7. Further, chronic cycled ethanol from 4 days in vitro (DIV) to 16DIV caused immediate 2-fold inductions of TNFα and IL-1β that grew to ~4-fold of age-matched control slices by 40DIV. This persistent inflammatory gene expression was completely reversed by microglial depletion and repopulation after chronic ethanol.ConclusionsMicroglia in OHSCs mediate proinflammatory responses to TLR agonists and ethanol. Microglial repopulation promoted an anti-inflammatory, trophic neuroenvironment and normalized proinflammatory gene expression. This supports the possibility of microglial depletion with repopulation as a strategy to reverse chronic neuroimmune activation.

Numerous natural products originated from Chinese herbal medicine exhibit anti-cancer activities, including anti-proliferative, pro-apoptotic, anti-metastatic, anti-angiogenic effects, as well as regulate autophagy, reverse multidrug resistance, balance immunity, and enhance chemotherapy in vitro and in vivo. To provide new insights into the critical path ahead, we systemically reviewed the most recent advances (reported since 2011) on the key compounds with anti-cancer effects derived from Chinese herbal medicine (curcumin, epigallocatechin gallate, berberine, artemisinin, ginsenoside Rg3, ursolic acid, silibinin, emodin, triptolide, cucurbitacin B, tanshinone I, oridonin, shikonin, gambogic acid, artesunate, wogonin, β-elemene, and cepharanthine) in scientific databases (PubMed, Web of Science, Medline, Scopus, and Clinical Trials). With a broader perspective, we focused on their recently discovered and/or investigated pharmacological effects, novel mechanism of action, relevant clinical studies, and their innovative applications in combined therapy and immunomodulation. In addition, the present review has extended to describe other promising compounds including dihydroartemisinin, ginsenoside Rh2, compound K, cucurbitacins D, E, I, tanshinone IIA and cryptotanshinone in view of their potentials in cancer therapy. Up to now, the evidence about the immunomodulatory effects and clinical trials of natural anti-cancer compounds from Chinese herbal medicine is very limited, and further research is needed to monitor their immunoregulatory effects and explore their mechanisms of action as modulators of immune checkpoints.

In order to investigate the impact of continuous dry heating treatment (CDHT) and repeated dry heating treatment (RDHT) on the structural and physicochemical properties of the waxy corn starch, the starch was prepared at 140 °C for continuous 20 h or for 5 cycles of heating, respectively. Each cycle lasted 4 h and the interval between each cycle was 1 h of cooling at room temperature. It turned out that RDHT and CDHT caused the aggregation of the starch into big lumps. The crystal type of CDHT and RDHT starches remained A-type, while the relative crystallinity and the short-range molecular order of starch characterized with 1040 cm−1/1018 cm−1 increased during first 2 circulations of RDHT and first 8 h of CDHT afterwards decreased upon the increased duration. After the heat treatment the water absorption index, swelling volume, viscosity and thermal parameters of CDHT and RDHT starches decreased, but the solubility, paste clarity and paste temperature of starches increased compared with native starch. Taken together RDHT has a greater impact on decreasing transmittance of starch paste, weakening the resistance to shear force and increasing retrogradation degree of starch paste compared to CDHT for the same duration.