Computer-assisted decision support for the usage of preventive antibacterial therapy in children with febrile pyelonephritis: A preliminary study.

Urinary tract infection (UTI) is one of the most common infectious diseases among children, but there is controversy regarding the use of preventive antibiotics for children first diagnosed with febrile pyelonephritis. To the best of our knowledge, no studies have addressed this issue by the deep learning technology. Therefore, in the current study, we conducted a study using Tc99m-DMSA renal static imaging data to investigate the need for preventive antibiotics on children first diagnosed with febrile pyelonephritis under 2 years old. The self-collected dataset comprised 64 children who did not require preventive antibiotic treatments and 112 children who did. Using several classic deep learning models, we verified that it is feasible to screen whether the first diagnosed children with febrile pyelonephritis require preventive antibacterial therapy, achieving a graded diagnosis. With the AlexNet model, we obtained accuracy of 84.05%, sensitivity of 81.71% and specificity of 86.70%, respectively. The experimental results indicate that deep learning technology could provide a new avenue to implement computer-assisted decision support for the diagnosis of the febrile pyelonephritis.

99mTc-3PRGD2 SPECT/CT Imaging for Diagnosing Lymph Node Metastasis of Primary Malignant Lung Tumors.

OBJECTIVE: To evaluate 99mtechnetium-three polyethylene glycol spacers-arginine-glycine-aspartic acid (99mTc-3PRGD2) single-photon emission computed tomography (SPECT)/computed tomography (CT) imaging for diagnosing lymph node metastasis of primary malignant lung neoplasms. MATERIALS AND METHODS: We prospectively enrolled 26 patients with primary malignant lung tumors who underwent 99mTc-3PRGD2 SPECT/CT and 18F-fluorodeoxyglucose (18F-FDG) positron emission tomography (PET)/CT imaging. Both imaging methods were analyzed in qualitative (visual dichotomous and 5-point grades for lymph nodes and lung tumors, respectively) and semi-quantitative (maximum tissue-to-background radioactive count) manners for the lymph nodes and lung tumors. The performance of the differentiation of lymph nodes with and without metastasis was determined at the per-lymph node station and per-patient levels using histopathological results as the reference standard. RESULTS: Total 42 stations had metastatic lymph nodes and 136 stations had benign lymph nodes. The differences between metastatic and benign lymph nodes in the visual qualitative and semiquantitative analyses of 99mTc-3PRGD2 SPECT/CT and 18F-FDG PET/CT were statistically significant (all P < 0.001). The area under the receiver operating characteristic curve (AUC) in the semi-quantitative analysis of 99mTc-3PRGD2 SPECT/CT was 0.908 (95% confidence interval [CI], 0.851-0.966), and the sensitivity, specificity, positive predictive value, and negative predictive value were 0.86 (36/42), 0.88 (120/136), 0.69 (36/52), and 0.95 (120/126), respectively. Among the 26 patients (including two patients each with two lung tumors), 15 had pathologically confirmed lymph node metastasis. The difference between primary lung lesions in patients with and without lymph node metastasis was statistically significant only in the semi-quantitative analysis of 99mTc-3PRGD2 SPECT/CT (P = 0.007), with an AUC of 0.807 (95% CI, 0.641-0.974). CONCLUSION: 99mTc-3PRGD2 SPECT/CT imaging may notably perform in the direct diagnosis of lymph node metastasis of primary malignant lung tumors and indirectly predict the presence of lymph node metastasis through uptake in the primary lesions.

Defects in Mitochondrial Biogenesis Drive Mitochondrial Alterations in PINK1-deficient Human Dopamine Neurons.

Mutations and loss of activity in the protein kinase PINK1 play a role in the pathogenesis of Parkinson's disease (PD). PINK1 regulates many aspects of mitochondrial quality control including mitochondrial autophagy (mitophagy), fission, fusion, transport, and biogenesis. Defects in mitophagy are though to play a predominant role in the loss of dopamine (DA) neurons in PD. Here we show that, although there are defects in mitophagy in human DA neurons lacking PINK1, mitochondrial deficits induced by the absence of PINK1 are primarily due to defects in mitochondrial biogenesis. Upregulation of PARIS and the subsequent down regulation of PGC-1a accounts for the mitochondrial biogenesis defects. CRISPR/Cas9 knockdown of PARIS completely restores the mitochondrial biogenesis defects and mitochondrial function without impacting the deficits in mitophagy due to the absence of PINK1. These results highlight the importance mitochondrial biogenesis in the pathogenesis of PD due to inactivation or loss of PINK1 in human DA neurons.

Diagnostic value of dynamic 18F-FDG PET/CT imaging in non-small cell lung cancer and FDG hypermetabolic lymph nodes.

Background: We aimed to investigate the diagnostic value of dynamic 2-deoxy-2-[18F]-fluoro-D-glucose positron emission tomography-computed tomography in non-small cell lung cancer and fluoro-D-glucose hypermetabolic lymph nodes. Methods: Patients who made an active appointment for positron emission tomography-computed tomography were randomly enrolled by referring to previous imaging data and clinical information. Finally, 34 histopathologically confirmed non-small cell lung cancers (18 adenocarcinoma and 16 squamous cell carcinoma cases) were prospectively studied using dynamic and static 2-deoxy-2-[18F]-fluoro-D-glucose positron emission tomography-computed tomography imaging (the diagnostic study has not yet been registered on a clinical trial platform). In dynamic positron emission tomography images, a volume of interest, defined by the thoracic aorta, was selected for estimating the arterial input function. Patlak and irreversible two-tissue compartment model analyses were performed based on the pixel points to obtain first-order characteristic kinetic parameters for each lesion and hypermetabolic lymph node. The first-order characteristic kinetic parameters were obtained based on the basic data of dynamic positron emission tomography images in the corresponding model and the lesion delineation of region-of-interest based on computed tomography images, such as V_Median (the median gray intensity of V), k3_Entropy, VB_Entropy, K1_Uniformity, and ki_Uniformity. The first-order characteristic kinetic parameters were also modeled by logistic regression for the differential diagnosis of non-small cell lung cancer and hypermetabolic lymph nodes. Maximum and mean standard uptake values (SUVmax and SUVmean, respectively) were obtained from static positron emission tomography images. The diagnostic efficacy of the parameters was evaluated using the receiver operating characteristic curve and the DeLong test. Results: There was a significant difference in the V_Median values of adenocarcinoma and squamous cell carcinoma. The regression models for K1, k3, and V provided good predictions of adenocarcinoma and squamous cell carcinoma typology. Significant differences were observed in k3_Entropy, VB_Entropy, K1_Uniformity, and ki_Uniformity between benign and malignant lymph nodes. The regression model of Ki, VB, and k3 could make a good prediction for identifying benign and malignant lymph nodes. Conclusions: Dynamic 2-deoxy-2-[18F]-fluoro-D-glucose positron emission tomography-computed tomography imaging showed high diagnostic value in the staging of non-small cell lung cancer and fluoro-D-glucose hypermetabolic lymph nodes, and can be of great use in non-small cell lung cancer lymph node staging and surgical decision-making.

Emerging Landscape of SARS-CoV-2 Variants and Detection Technologies.

In 2019, a new coronavirus was identified that has caused significant morbidity and mortality worldwide. Like all RNA viruses, severe acute respiratory syndrome coronavirus 2 (SARS-Cov-2) evolves over time through random mutation resulting in genetic variations in the population. Although the currently approved coronavirus disease 2019 vaccines can be given to those over 5 years of age and older in most countries, strikingly, the number of people diagnosed positive for SARS-Cov-2 is still increasing. Therefore, to prevent and control this epidemic, early diagnosis of infected individuals is of great importance. The current detection of SARS-Cov-2 coronavirus variants are mainly based on reverse transcription-polymerase chain reaction. Although the sensitivity of reverse transcription-polymerase chain reaction is high, it has some disadvantages, for example, multiple temperature changes, long detection time, complicated operation, expensive instruments, and the need for professional personnel, which brings considerable inconvenience to the early diagnosis of this virus. This review comprehensively summarizes the development and application of various current detection technologies for novel coronaviruses, including isothermal amplification, CRISPR-Cas detection, serological detection, biosensor, ensemble, and microfluidic technology, along with next-generation sequencing. Those findings offer us a great potential to replace or combine with reverse transcription-polymerase chain reaction detection to achieve the purpose of allowing predictive diagnostics and targeted prevention of SARS-Cov-2 in the future.

A nociceptive neuronal ensemble in the dorsomedial prefrontal cortex underlies pain chronicity.

Pain chronicity involves unpleasant experience in both somatosensory and affective aspects, accompanied with the prefrontal cortex (PFC) neuroplastic alterations. However, whether specific PFC neuronal ensembles underlie pain chronicity remains elusive. Here we identify a nociceptive neuronal ensemble in the dorsomedial prefrontal cortex (dmPFC), which shows prominent reactivity to nociceptive stimuli. We observed that this ensemble shows distinct molecular characteristics and is densely connected to pain-related regions including basolateral amygdala (BLA) and lateral parabrachial nuclei (LPB). Prolonged chemogenetic activation of this nociceptive neuronal ensemble, but not a randomly transfected subset of dmPFC neurons, induces chronic pain-like behaviors in normal mice. By contrast, silencing the nociceptive dmPFC neurons relieves both pain hypersensitivity and anxiety in mice with chronic inflammatory pain. These results suggest the presence of specific dmPFC neuronal ensembles in processing nociceptive information and regulating pain chronicity.

The use of three-dimensional endoscope in transnasal skull base surgery: A single-center experience from China.

Objective: The development of skull base surgery in the past decade has been influenced by advances in visualization techniques; recently, due to such improvements, 3D endoscopes have been widely used. Herein, we address its effect for transnasal endoscopic skull base surgery. Methods: A total of 63 patients who under endoscopic endonasal surgery (EES) with 3-D endoscope were retrospectively reviewed, including pituitary adenomas, craniopharyngiomas, meningiomas, Rathke's cleft cysts, and chordomas. According to different lesions, transsellar approach (24 cases), transsphenoidal-transtuberculum approach (14 cases), transclival approach (6 cases), and transpterygoid approach (19 cases) were selected. Results: Total removal of tumors was achieved in 56 patients (88.9%) and subtotal removal in 7 cases (11.1%). Complications included diabetes insipidus in seven patients (11.1%), cerebrospinal fluid (CSF) leakage in two patients (3.2%), major vascular injury occurred in one patient (1.6%), cranial nerve injury in nine patients (14.3%), and meningitis in two patients (3.2%). There was no mortality in the series. All patients recovered and were back to normal daily life, and no tumor recurrence or delayed CSF leakage was detected during the follow-up (2-13 months, mean 7.59 months). Conclusions: Via 3D EES, it improved depth perception and preserved important neurovascular tissue when tumors were removed, which is important for improving the operative prognosis.

Exploration of the causes of cerebrospinal fluid leakage after endoscopic endonasal surgery for sellar and suprasellar lesions and analysis of risk factors.

Objective: Postoperative cerebrospinal fluid (CSF) leakage following endoscopic endonasal surgery (EES) is a frequent complication. This study aims to identify potential risk factors of postoperative CSF leakage. Methods: A retrospective review of 360 patients who underwent EES was included. The associations between postoperative CSF leakage and patient demographics, medical history, tumor characteristics, and intraoperative repair techniques were analyzed; the diagnosis and repair of postoperative CSF leakage were also introduced. Results: Postoperative CSF leakage occurred in 14 patients (3.9%), 2 of them cured by lumbar cistern drainage, 12 underwent endoscopic repair. Among these 12 cases, 3 were repaired twice, and the rest were cured the first time. During the repair surgery, insufficient embedded fat was detected in one case detected, seven with breached inner artificial dura, three had vascularized pedicle nasoseptal flap (VP-NSF) displacement, two with VP-NSF perforation, two with VP-NSF inactivation, and one with imperfect adherence to VP-NSF to the skull base. Eight cases had intracranial infections. Excluding one case who died of severe intracranial infection, the rest were cured and discharged without obvious sequelae. Multivariate analysis revealed that the suprasellar lesion, subarachnoid invasion, and intraoperative grade 3 flow CSF leakage were the risk factors of CSF leakage after operation, while the bone flap was a protective factor. Conclusion: Bone flap combined with VP-NSF and iodoform gauze for skull base reconstruction is recommended in high-risk patients, while postoperative lumbar cistern drain remains dispensable.

Advances in Clinical Oncology Research on 99mTc-3PRGD2 SPECT Imaging.

The integrin alpha(α)v beta(ß)3 receptor is ubiquitous in malignant tumors and has a certain level of specificity for tumors. Technetium-99m hydrazinonicotinamide-dimeric cyclic arginyl-glycyl-aspartic acid peptide with three polyethylene glycol spacers (99mTc-3PRGD2) can bind specifically to the integrin αvß3 receptor with high selectivity and strong affinity. Thus, it can specifically mark tumors and regions with angiogenesis for tumor detection and be used in single-photon emission computed tomography (SPECT) imaging. This modality has good application value for diagnosing and treating tumor lesions, such as those in the lung, breast, esophagus, head, and neck. This review provides an overview of the current clinical research progress of 99mTc-3PRGD2 SPECT imaging for tumor lesions, including for the diagnosis and differential diagnosis of tumors in different body parts, evaluation of related metastases, and evaluation of efficacy. In addition, the future clinical application prospects and possibilities of 99mTc-3PRGD2 SPECT imaging are further discussed.

AttCRISPR: a spacetime interpretable model for prediction of sgRNA on-target activity.

BACKGROUND: More and more Cas9 variants with higher specificity are developed to avoid the off-target effect, which brings a significant volume of experimental data. Conventional machine learning performs poorly on these datasets, while the methods based on deep learning often lack interpretability, which makes researchers have to trade-off accuracy and interpretability. It is necessary to develop a method that can not only match deep learning-based methods in performance but also with good interpretability that can be comparable to conventional machine learning methods. RESULTS: To overcome these problems, we propose an intrinsically interpretable method called AttCRISPR based on deep learning to predict the on-target activity. The advantage of AttCRISPR lies in using the ensemble learning strategy to stack available encoding-based methods and embedding-based methods with strong interpretability. Comparison with the state-of-the-art methods using WT-SpCas9, eSpCas9(1.1), SpCas9-HF1 datasets, AttCRISPR can achieve an average Spearman value of 0.872, 0.867, 0.867, respectively on several public datasets, which is superior to these methods. Furthermore, benefits from two attention modules-one spatial and one temporal, AttCRISPR has good interpretability. Through these modules, we can understand the decisions made by AttCRISPR at both global and local levels without other post hoc explanations techniques. CONCLUSION: With the trained models, we reveal the preference for each position-dependent nucleotide on the sgRNA (short guide RNA) sequence in each dataset at a global level. And at a local level, we prove that the interpretability of AttCRISPR can be used to guide the researchers to design sgRNA with higher activity.

IDH1 mutation contributes to myeloid dysplasia in mice by disturbing heme biosynthesis and erythropoiesis.

Isocitrate dehydrogenase (IDH) mutations are common genetic alterations in myeloid disorders, including acute myeloid leukemia (AML) and myelodysplastic syndrome (MDS). Epigenetic changes, including abnormal histone and DNA methylation, have been implicated in the pathogenic build-up of hematopoietic progenitors, but it is still unclear whether and how IDH mutations themselves affect hematopoiesis. Here, we show that IDH1-mutant mice develop myeloid dysplasia in that these animals exhibit anemia, ineffective erythropoiesis, and increased immature progenitors and erythroblasts. In erythroid cells of these mice, D-2-hydroxyglutarate, an aberrant metabolite produced by the mutant IDH1 enzyme, inhibits oxoglutarate dehydrogenase activity and diminishes succinyl-coenzyme A (CoA) production. This succinyl-CoA deficiency attenuates heme biosynthesis in IDH1-mutant hematopoietic cells, thus blocking erythroid differentiation at the late erythroblast stage and the erythroid commitment of hematopoietic stem cells, while the exogenous succinyl-CoA or 5-ALA rescues erythropoiesis in IDH1-mutant erythroid cells. Heme deficiency also impairs heme oxygenase-1 expression, which reduces levels of important heme catabolites such as biliverdin and bilirubin. These deficits result in accumulation of excessive reactive oxygen species that induce the cell death of IDH1-mutant erythroid cells. Our results clearly show the essential role of IDH1 in normal erythropoiesis and describe how its mutation leads to myeloid disorders. These data thus have important implications for the devising of new treatments for IDH-mutant tumors.

Isocitrate dehydrogenase 1 mutation enhances 24(S)-hydroxycholesterol production and alters cholesterol homeostasis in glioma.

Isocitrate dehydrogenase (IDH) mutation is the most important initiating event in gliomagenesis, and the increasing evidence shows that IDH mutation is associated with the metabolic reprogramming in the tumor. Dysregulated cholesterol metabolism is a hallmark of tumor cells, but the cholesterol homeostasis in IDH-mutated glioma is still unknown. In this study, we found that astrocyte-specific mutant IDH1(R132H) knockin reduced the cholesterol contents and damaged the structure of myelin in mouse brains. In U87 and U251 cells, the expression of mutant IDH1 consistently reduced the cholesterol levels. Furthermore, we found that IDH1 mutation enhanced the production of 24(S)-hydroxycholesterol (24-OHC), which is not only the metabolite of cholesterol elimination, but also functions as an endogenous ligand for the liver X receptors (LXRs). In IDH1-mutant glioma cells, the elevated 24-OHC activated LXRs, which consequently accelerated the low-density lipoprotein receptor (LDLR) degradation by upregulating the inducible degrader of the LDLR (IDOL). The reduced LDLR expressions in IDH1-mutant glioma cells abated the uptakes of low-density lipoprotein (LDL) to decrease the cholesterol influx. In addition, the activated LXRs also promoted the cholesterol efflux by elevating the ATP-binding cassette transporter A1 (ABCA1), ABCG1, and apolipoprotein E (ApoE) in both IDH1-mutant astrocytes and glioma cells. As a feedback, the reduced cholesterol levels stimulated the cholesterol biosynthesis, which made IDH1-mutated glioma cells more sensitive to atorvastatin, an inhibitor of 3-hydroxy-3-methylglutaryl-CoA reductase. The altered cholesterol homeostasis regulated by mutant IDH provides a pivotal therapeutical strategy for the IDH-mutated gliomas.

Wild-Type IDH1 and Mutant IDH1 Opposingly Regulate Podoplanin Expression in Glioma.

Isocitrate dehydrogenase (IDH) mutations occur frequently in lower-grade gliomas, which result in genome-wide epigenetic alterations. The wild-type IDH1 is reported to participate in lipid biosynthesis and amino acid metabolism, but its role in tumorigenesis is still unclear. In this study, the expressions of IDH1 and podoplanin (Pdpn) were determined in IDH-mutated and IDH-wild-type gliomas, and their relationships in glioma were further analyzed. In addition, the regulation of wild-type IDH1 and mutant IDH1 on Pdpn expression was investigated by luciferase assays and promoter methylation analysis. Our study showed that Pdpn was almost undetectable in IDH-mutated glioma but strongly expressed in higher-grade IDH-wild-type glioma. Pdpn overexpression promoted the migration of glioma cells but had little effect on cell growth. Moreover, Pdpn expression was positively correlated with the increased wild-type IDH1 levels in IDH-wild-type glioma. Consistently, the wild-type IDH1 greatly promoted the transcription and expression of Pdpn, but the mutant IDH1 and D-2-hydroxyglutarate significantly suppressed Pdpn expression in glioma cells. Besides, our results revealed that the methylation of CpG islands in the Pdpn promoter was opposingly regulated by wild-type and mutant IDH1 in glioma. Collectively, our results indicated that wild-type and mutant IDH1 opposingly controlled the Pdpn expression in glioma by regulating its promoter methylation, which provides a basis for understanding the relationship between wild-type and mutant IDH1 in epigenetic regulation and tumorigenesis.

A comparative study on the mechanical and reactive behavior of three fluorine-containing thermites.

Thermite serves as a kind of representative energetic material, which is extensively applied in the civil and military fields. In this paper, PTFE/Al/Fe2O3, PTFE/Al/MnO2 and PTFE/Al/MoO3, solid fluorine-containing thermite with different PTFE content, were successfully fabricated by referring to the traditional thermite and adding PTFE as a binder or matrix. Quasi-static compression tests were performed to investigate the mechanical and reactive behavior of fluorine-containing thermite. SEM and XRD were employed to analyze and characterize the energetic composites and reaction residuals. The results show that all types of fluorine-containing thermite exhibited different mechanical behavior. PTFE/Al/MnO2 exhibited the lowest yield strength and strain hardening modulus, but the highest compressive strength and toughness. With the increase of PTFE content, the strength of fluorine-containing thermite improved. No reaction occurred when the PTFE content was 60 vol%, while fluorine-containing thermite with a PTFE content of 80 vol% experienced a severe exothermic reaction under quasi-static compression. The ignition of PTFE/Al/MoO3 and PTFE/Al/Fe2O3 actually attributed to the reaction of Al and PTFE, and the reaction between Al and Fe2O3 or MoO3 was not excited due to the insufficient input energy. The thermite reaction between Al and MnO2, as well as the reaction of MnO2 and PTFE, was induced because PTFE/Al/MnO2 possessed excellent ductility and absorbed the most energy during compression, accompanied with the production of Mn and MnF2.

Plin5 deficiency exacerbates pressure overload-induced cardiac hypertrophy and heart failure by enhancing myocardial fatty acid oxidation and oxidative stress.

While cardiac hypertrophy and heart failure are accompanied by significant alterations in energy metabolism, more than 50-70% of energy is obtained from fatty acid ß-oxidation (FAO) in adult hearts under physiological conditions. Plin5 is involved in the metabolism of lipid droplets (LDs) and is highly abundant in oxidative tissues including heart, liver and skeletal muscle. Plin5 protects the storage of triglyceride (TG) in LDs by inhibiting lipolysis, thereby suppressing excess FAO and preventing excessive oxidative stress in the heart. In this study, we investigated the roles of Plin5 in cardiac hypertrophy and heart failure in mice treated with transverse aortic constriction (TAC). The results indicated that Plin5 deficiency aggravated myocardial hypertrophy in the TAC-treated mice and exacerbated the TAC-induced heart failure. We also found that Plin5 deficiency reduced the cardiac lipid accumulation and upregulated the levels of PPARα and PGC-1α, which stimulate mitochondrial proliferation. Moreover, Plin5 deficiency aggravated the TAC-induced oxidative stress. We consistently found that Plin5 knockdown disrupted TG storage and elevated FAO and lipolysis in H9C2 rat cardiomyocytes. In addition, Plin5 knockdown also provoked mitochondrial proliferation and lipotoxic injury in H9C2 cells. In conclusion, Plin5 deficiency increases myocardial lipolysis, elevates FAO and oxidative burden, and thereby exacerbates cardiac hypertrophy and heart failure in TAC-treated mice.

The novel gene TRIM44L from orange-spotted grouper negatively regulates the interferon response.

Accumulated evidence suggests that some of the tripartite motif (TRIM) -family proteins function as critical regulators of carcinogenesis, immunity, and antiviral functions. TRIM44 is an atypical TRIM family protein that lacks the entire RING domain and has been demonstrated to play a crucial role in cancer and viral infection. To our knowledge, the role of TRIM44 in fish still remains largely unknown. Here, we cloned and characterized a novel TRIM44-like gene from orange spotted grouper (EcTRIM44L). Sequence analysis indicated that EcTRIM44L encoded a 393 amino acid peptide, which shared 81.44% and 51.02% identity with large yellow croaker (Larimichthys crocea) and zebrafish (Danio rerio), respectively. However, EcTRIM44L only exhibited 24.69% identity with the TRIM44 protein of humans (Homo sapiens). Moreover, EcTRIM44L contained two conserved domains, including a B-Box domain and a coiled-coil domain, but not a RING domain. Using fluorescence microscopy, we observed green fluorescence in the cytoplasm of the EcTRIM44L-EGFP transfected grouper spleen (GS) cells. As the infection proceeded, EcTRIM44L transcription was significantly up-regulated in red-spotted grouper nervous necrosis virus (RGNNV) infection, suggesting that EcTRIM44L might be involved in fish virus infections. The in vitro overexpression of EcTRIM44L significantly enhanced RGNNV replication, as demonstrated by the accelerated cytopathic effect (CPE) progression induced by RGNNV, as well as the increased expression of coat protein (CP) and RNA-dependent RNA polymerase (RdRp). The overexpression of EcTRIM44L significantly decreased the level of interferon (IFN) related signaling molecules and pro-inflammatory cytokine expression, suggesting that EcTRIM44L affected virus replication by negatively regulating the IFN response. In addition, the melanoma differentiation-associated protein 5 (MDA5) and mitochondrial antiviral-signaling protein (MAVS), but not mediator of IRF3 activation (MITA)-evoked IFN response was negatively regulated by EcTRIM44L. Together, for the first time, our results indicate that EcTRIM44L negatively regulates the interferon response against grouper RNA virus infection.

Plin3 protects against alcoholic liver injury by facilitating lipid export from the endoplasmic reticulum.

Hepatic lipid accumulation is the most common pathological characteristic of alcoholic liver disease (ALD). In mammalian cells, excess neutral lipids are stored in lipid droplets (LDs). As a member of perilipin family proteins, Plin3 was recently found to regulate the LD biogenesis. However, the roles and mechanism of Plin3 in ALD progression remain unclear. Herein, we found that alcohol stimulated Plin3 expression in both mouse livers and cultured AML12 mouse hepatic cells, which was accompanied by excess LD accumulation in hepatocytes. The elevations of Plin3 in alcohol-treated hepatocytes paralleled with the levels of both PPARα and γ, and the protein degradation of Plin3 was also reduced after alcohol exposure. Moreover, Plin3 knockdown increased cellular sensitivity to alcohol-induced apoptosis, endoplasmic reticulum (ER) stress, and inflammatory cytokines release, including TNF-α, IL-1, and IL-6ß. Notably, alcohol exacerbated triglycerides (TG) accumulation in the ER and caused ER dilation in Plin3-knockdown AML12 cells. Finally, we observed that Plin3 interacted with dynein subunit Dync1i1 and mediated the colocalization of LDs and microtubules, while high concentration of alcohol disrupted microtubules and caused dispersion of excess small LDs in cytoplasm. Summarily, Plin3 promotes lipid export from the ER and reduces ER lipotoxic stress, thereby, protecting against alcoholic liver injury. Moreover, Plin3 could be an adapter protein mediating LD transport by microtubules. This study explored the roles of Plin3 in alcohol-induced hepatic injury, suggesting Plin3 as a potential target for the prevention of ALD progression.

Perilipin 5 alleviates HCV NS5A-induced lipotoxic injuries in liver.

BACKGROUND: The homeostasis of lipid droplets (LDs) plays a crucial role in maintaining the physical metabolic processes in cells, and is regulated by many LD-associated proteins, including perilipin 5 (Plin5) in liver. As the putative sites of hepatitis C virus (HCV) virion assembly, LDs are vital to viral infection. In addition, the hepatic LD metabolism can be disturbed by non-structural HCV proteins, such as NS5A, but the details are still inexplicit. METHODS: HCV NS5A was overexpressed in the livers and hepatocytes of wild-type and Plin5-null mice. BODIPY 493/503 and oil red O staining were used to detect the lipid content in mouse livers and hepatocytes. The levels of lipids, lipid peroxidation and inflammation biomarkers were further determined. Immunofluorescence assay and co-immunoprecipitation assay were performed to investigate the relationship of Plin5 and NS5A. RESULTS: One week after adenovirus injection, livers expressing NS5A showed more inflammatory cell aggregation and more severe hepatic injuries in Plin5-null mice than in control mice, which was consistent with the increased serum levels of IL-2 and TNF-α (P < 0.05) observed in Plin5-null mice. Moreover, Plin5 deficiency in the liver and hepatocytes aggravated the elevation of MDA and 4-HNE levels induced by NS5A expression (P < 0.01). The triglyceride (TG) content was increased approximately 25% by NS5A expression in the wild-type liver and hepatocytes but was unchanged in the Plin5-null liver and hepatocytes. More importantly, Plin5 deficiency in the liver and hepatocytes exacerbated the elevation of non-esterified fatty acids (NEFAs) stimulated by NS5A expression (P < 0.05 and 0.01 respectively). Using triacsin C to block acyl-CoA biosynthesis, we found that Plin5 deficiency aggravated the NS5A-induced lipolysis of TG. In contrast, Plin5 overexpression in HepG2 cells ameliorated the NS5A-induced lipolysis and lipotoxic injuries. Immunofluorescent staining demonstrated that NS5A expression stimulated the targeting of Plin5 to the surface of the LDs in hepatocytes without altering the protein levels of Plin5. By co-IP, we found that the N-terminal domain (aa 32-128) of Plin5 was pivotal for its binding with NS5A. CONCLUSIONS: Our data highlight a protective role of Plin5 against hepatic lipotoxic injuries induced by HCV NS5A, which is helpful for understanding the steatosis and injuries in liver during HCV infection.

Isocitrate dehydrogenase1 mutation reduces the pericyte coverage of microvessels in astrocytic tumours.

INTRODUCTION: Tumour-associated angiogenesis is associated with the malignancy and poor prognosis of glioma. Isocitrate dehydrogenase (IDH) mutations are present in the majority of lower-grade (WHO grade II and III) and secondary glioblastomas, but their roles in tumour angiogenesis remain unclear. METHODS: Using magnetic resonance imaging (MRI), the cerebral blood flow (CBF) of IDH-mutated glioma was measured and compared with the IDH-wildtype glioma. The densities of microvessels in IDH-mutated and wildtype astrocytoma and glioblastoma were assessed by immunohistochemical (IHC) staining with CD34, and the pericytes were labelled with α-smooth muscle antigen (α-SMA), neural-glial antigen 2 (NG2) and PDGF receptor-ß (PDGFR-ß), respectively. Furthermore, glia-specific mutant IDH1 knock-in mice were generated to evaluate the roles of mutant IDH1 on brain vascular architectures. The transcriptions of the angiogenesis-related genes were assessed in TCGA datasets, including ANGPT1, PDGFB and VEGFA. The expressions of these genes were further determined by western blot in U87-MG cells expressing a mutant IDH1 or treated with 2-HG. RESULTS: The MRI results indicated that CBF was reduced in the IDH-mutated gliomas. The IHC staining showed that the pericyte coverages of microvessels were significantly decreased, but the microvessel densities (MVDs) were only slightly decreased in IDH-mutated glioma. The mutant IDH1 knock-in also impeded the pericyte coverage of brain microvessels in mice. Moreover, the TCGA database showed the mRNA levels of angiogenesis factors, including ANGPT1, PDGFB and VEGFA, were downregulated, and their promoters were also highly hyper-methylated in IDH-mutated gliomas. In addition, both mutant IDH1 and D-2-HG could downregulate the expression of these genes in U87-MG cells. CONCLUSIONS: Our results suggested that IDH mutations could reduce the pericyte coverage of microvessels in astrocytic tumours by inhibiting the expression of angiogenesis factors. As vascular pericytes play an essential role in maintaining functional blood vessels to support tumour growth, our findings imply a potential avenue of therapeutic strategy for IDH-mutated gliomas.

Landslide Susceptibility Assessment Using Integrated Deep Learning Algorithm along the China-Nepal Highway.

The China-Nepal Highway is a vital land route in the Kush-Himalayan region. The occurrence of mountain hazards in this area is a matter of serious concern. Thus, it is of great importance to perform hazard assessments in a more accurate and real-time way. Based on temporal and spatial sensor data, this study tries to use data-driven algorithms to predict landslide susceptibility. Ten landslide instability factors were prepared, including elevation, slope angle, slope aspect, plan curvature, vegetation index, built-up index, stream power, lithology, precipitation intensity, and cumulative precipitation index. Four machine learning algorithms, namely decision tree (DT), support vector machines (SVM), Back Propagation neural network (BPNN), and Long Short Term Memory (LSTM) are implemented, and their final prediction accuracies are compared. The experimental results showed that the prediction accuracies of BPNN, SVM, DT, and LSTM in the test areas are 62.0%, 72.9%, 60.4%, and 81.2%, respectively. LSTM outperformed the other three models due to its capability to learn time series with long temporal dependencies. It indicates that the dynamic change course of geological and geographic parameters is an important indicator in reflecting landslide susceptibility.