The APSES Transcription Factor SsStuA Regulating Cell Wall Integrity Is Essential for Sclerotia Formation and Pathogenicity in Sclerotinia sclerotiorum.

APSES (Asm1p, Phd1p, Sok2p, Efg1p, and StuAp) family transcription factors play crucial roles in various biological processes of fungi, however, their functional characterization in phytopathogenic fungi is limited. In this study, we explored the role of SsStuA, a typical APSES transcription factor, in the regulation of cell wall integrity (CWI), sclerotia formation and pathogenicity of Sclerotinia sclerotiorum, which is a globally important plant pathogenic fungus. A deficiency of SsStuA led to abnormal phosphorylation level of SsSmk3, the key gene SsAGM1 for UDP-GlcNAc synthesis was unable to respond to cell wall stress, and decreased tolerance to tebuconazole. In addition, ΔSsStuA was unable to form sclerotia but produced more compound appressoria. Nevertheless, the virulence of ΔSsStuA was significantly reduced due to the deficiency of the invasive hyphal growth and increased susceptibility to hydrogen peroxide. We also revealed that SsStuA could bind to the promoter of catalase family genes which regulate the expression of catalase genes. Furthermore, the level of reactive oxygen species (ROS) accumulation was found to be increased in ΔSsStuA. In summary, SsStuA, as a core transcription factor involved in the CWI pathway and ROS response, is required for vegetative growth, sclerotia formation, fungicide tolerance and the full virulence of S. sclerotiorum.

The study on mechanical model considering optimal self-adaption in the bottleneck area.

It aims to solve the problem that the evacuation state of pedestrians depicted by the traditional social force model in a crowded multiexit scenario has a relatively large difference with the actual state, especially the 'optimal path' considered by the self-driving force is the problem of shortest path, and the multiexit evacuation mode depicted by the 'herd behavior' is the local optimum problem. Through in-depth analysis of actual evacuation data of pedestrians and causes of problem, a new crowd evacuation optimization model is established in order to effectively improve the simulation accuracy of crowd evacuation in a multi-exit environment. The model obtains the direction of motion of pedestrians using a field model, fully considers the factors such as exit distance, distribution of pedestrians and regional crowding degree, makes a global optimization for the self-driving force in the social force model using a centralized and distributed network model, and makes a local optimization for it using an elephant herding algorithm, so as to establish a new evacuation optimization method for optimal self-adaption in the bottleneck area. The performance status is compared between the improved social force model and the new model by experiments, and the key factors that affect the new model are analyzed in an in-depth manner. The results show that the new model can optimize the optimal path choice at the early stage of evacuation and improve the evacuation efficiency of pedestrians at the late stage, so as to ensure relatively even distribution of pedestrians at each exit, and also make the simulated evacuation process be more real; and the improvement in overall evacuation efficiency is greater when the number of pedestrians to be evacuated is larger. Therefore, the new model provides a method to solve the phenomenon of disorder in overall pedestrian evacuation due to excessive crowd density during the process of multi-exit evacuation.

Kj-mhpC Enzyme in Klebsiella jilinsis 2N3 Is Involved in the Degradation of Chlorimuron-Ethyl via De-Esterification.

Microbial degradation is a highly efficient and reliable approach for mitigating the contamination of sulfonylurea herbicides, such as chlorimuron-ethyl, in soil and water. In this study, we aimed to assess whether Kj-mhpC plays a pivotal role in the degradation of chlorimuron-ethyl. Kj-mhpC enzyme purified via prokaryotic expression exhibited the highest catalytic activity for chlorimuron-ethyl at 35 °C and pH 7. Bioinformatic analysis and three-dimensional homologous modeling of Kj-mhpC were conducted. Additionally, the presence of Mg+ and Cu2+ ions partially inhibited but Pb2+ ions completely inhibited the enzymatic activity of Kj-mhpC. LC/MS revealed that Kj-mhpC hydrolyzes the ester bond of chlorimuron-ethyl, resulting in the formation of 2-(4-chloro-6-methoxypyrimidine-2-amidoformamidesulfonyl) benzoic acid. Furthermore, the point mutation of serine at position 67 (Ser67) confirmed that it is the key amino acid at the active site for degrading chlorimuron-ethyl. This study enhanced the understanding of how chlorimuron-ethyl is degraded by microorganisms and provided a reference for bioremediation of the environment polluted with chlorimuron-ethyl.

Experimental study of precursory features of CO2 blasting-induced coal rock fracture based on grayscale and texture analysis.

CO2 blasting has been identified as a potent method for enhancing the permeability of coal seams and improving gas drainage efficiency. This study is focused on elucidating the deformation and fracture mechanisms of coal and rock during CO2 blasting and on identifying the precursor characteristics of these processes. To this end, a CO2 blasting-induced coal rock fracture pressure model and a gas pressure distribution model were developed. The research utilized a self-developed CO2 blasting test platform along with a non-contact full-strain field measurement analysis system. Briquette samples were subjected to CO2 blasting tests under controlled experimental conditions, which included an axial pressure of 1.0 MPa and variable gas pressures of 0.5, 1.0, and 1.5 MPa. This methodology enabled the capture of the principal strain field on the surface of the samples. The Gray Level Co-occurrence Matrix (GLCM) was employed to extract and analyze the grayscale and texture features of the strain cloud maps, facilitating a quantitative assessment of their evolution. The aim was to pinpoint the precursor characteristics associated with coal rock cracking and crack propagation. The results revealed that: (1) During the cracking and subsequent propagation of samples, the strain field's grayscale histogram underwent a transformation from a "broad and low" to a "narrow and high" configuration, with a consistent increase in peak frequency. Specifically, at 3 ms, a primary crack was observed in the sample, evidenced by a grayscale peak frequency of 0.0846. By 9 ms, as the crack propagated, the grayscale peak frequency escalated to 0.1626. (2) The texture feature parameters experienced their initial abrupt change at 3ms. Correlation with the gas pressure distribution model indicated that this was the crack initiation moment in the sample. (3) A secondary abrupt shift in the texture feature parameters occurred at 9ms, in conjunction with experimental phenomena, was identified as the crack propagation phase. Monitoring the grayscale and texture features of the principal strain field on the coal rock surface proved effective in recognizing the precursor characteristics of crack initiation and propagation. This research has the potential to reduce blasting costs in coal mines, optimize blasting effects, and provided theoretical guidance for enhancing gas extraction efficiency from deep and low permeability coal seams.

Molecular Mechanism Underlying Pathogenicity Inhibition by Chitosan in Cochliobolus heterostrophus.

Chitosan, as a natural nontoxic biomaterial, has been demonstrated to inhibit fungal growth and enhance plant defense against pathogen infection. However, the antifungal pattern and mechanism of how chitosan application evokes plant defense are poorly elucidated. Herein, we provide evidence that chitosan exposure is fungicidal to C. heterostrophus. Chitosan application impairs conidia germination and appressorium formation of C. heterostrophus and has a pronounced effect on reactive oxygen species production, thereby preventing infection in maize. In addition, the toxicity of chitosan to C. heterostrophus requires Mkk1 and Mps1, two key components in the cell wall integrity pathway. The Δmkk1 and Δmps1 mutants were more tolerant to chitosan than the wild-type. To dissect chitosan-mediated plant defense response to C. heterostrophus, we conducted a metabolomic analysis, and several antifungal compounds were upregulated in maize upon chitosan treatment. Taken together, our findings provide a comprehensive understanding of the mechanism of chitosan-alleviated infection of C. heterostrophus, which would promote the application of chitosan in plant protection in agriculture.

Transcriptomic analysis and knockout experiments reveal the role of suhB in the biocontrol effects of Pantoea jilinensis D25 on Botrytis cinerea.

Tomato gray mold, caused by Botrytis cinerea, is an important disease in tomato. Pantoea jilinensis D25, isolated form tomato rhizosphere soil, can prevent B. cinerea infection in tomato. To determine the underlying biocontrol mechanism, the transcriptome of P. jilinensis D25 was assessed. Differential expression analysis revealed that 941 genes were upregulated and 997 genes were downregulated. Through transcriptome analysis, the suhB gene was knocked out. ΔPj-suhB exhibited lower swimming motility and colonization abilities than strain D25. After 4 days of co-cultivation, ΔPj-suhB could reduce the colony diameter, mycelial weight, and spore production of B. cinerea with the inhibitory rates of 31.72 %, 39.62 %, and 47.42 %, respectively, compared with control. However, the inhibitory rates of strain D25 were 52.91 %, 60.09 %, and 76.85 %, respectively, compared with control. Strain D25 could significantly downregulate pathogenesis-related genes in B. cinerea, whereas the expression level of these genes in B. cinerea was higher after treatment with ΔPj-suhB than after that with strain D25. In vitro experiments revealed that the lesion area and disease control efficacy were 1.520 and 0.038 cm2 and 68.7 % and 99.0 %, respectively, after ΔPj-suhB and strain D25 treatments. Pot experiments revealed that ΔPj-suhB and strain D25 could prevent tomato plants from B. cinerea infection with the disease reduction rate of 37.5 % and 75.0 %, respectively. Though the activities of defense-related enzymes and expression level of defense related genes in tomato plants were increased under ΔPj-suhB treatment, these effects were higher after strain D25 treatment. Thus, these results demonstrated that suhB was the key gene in strain D25 underlying its biocontrol effect and mobility.

Spinal infection caused by Aspergillus flavus in a diabetic: a case report and literature review.

Spinal infections, notably those induced by Aspergillus flavus (A. flavus), represent a complex and uncommon clinical challenge. In individuals with diabetes mellitus, the risk is exacerbated due to a compromised immune response and a heightened vulnerability to non-standard pathogens. This case report chronicles the intricate diagnostic and treatment journey of a 59-year-old diabetic patient grappling with a spinal infection attributed to A. flavus. The diagnosis was delayed due to non-specific symptoms and unclear radiological signs. The administration of voriconazole, a targeted antifungal treatment, resulted in a significant clinical and radiological improvement, underscoring its effectiveness in treating such unusual fungal spinal infections; meanwhile, we found that terbinafine hydrochloride also has a similar effect in treating fungal spinal infections. This case underscores the importance of considering fungal causes in spinal infections among diabetic patients and highlights prompt diagnosis and individualized targeted antifungal therapy.

Infection-specific transcriptional patterns of the maize pathogen Cochliobolus heterostrophus unravel genes involved in asexual development and virulence.

Southern corn leaf blight (SCLB) caused by Cochliobolus heterostrophus is a destructive disease that threatens global maize (Zea mays) production. Despite many studies being conducted, very little is known about molecular processes employed by the pathogen during infection. There is a need to understand the fungal arms strategy and identify novel functional genes as targets for fungicide development. Transcriptome analysis based on RNA sequencing was carried out across conidia germination and host infection by C. heterostrophus. The present study revealed major changes in C. heterostrophus gene expression during host infection. Several differentially expressed genes (DEGs) induced during C. heterostrophus infection could be involved in the biosynthesis of secondary metabolites, peroxisome, energy metabolism, amino acid degradation and oxidative phosphorylation. In addition, histone acetyltransferase, secreted proteins, peroxisomal proteins, NADPH oxidase and transcription factors were selected for further functional validation. Here, we demonstrated that histone acetyltransferases (Hat2 and Rtt109), secreted proteins (Cel61A and Mep1), peroxisomal proteins (Pex11A and Pex14), NADPH oxidases (NoxA, NoxD and NoxR) and transcription factors (Crz1 and MtfA) play essential roles in C. heterostrophus conidiation, stress adaption and virulence. Taken together, our study revealed major changes in gene expression associated with C. heterostrophus infection and identified a diverse repertoire of genes critical for successful infection.

The histidine kinases regulate allyl-isothiocyanate sensitivity in Cochliobolus heterostrophus.

BACKGROUD: Two-component histidine kinase (HK) phosphorelay signaling systems play important roles in differentiation, virulence, secondary metabolite production and response to environmental signals. Allyl isothiocyanate (A-ITC) is a hydrolysis product of glucosinolates with excellent antifungal activity. Our previous study indicated that the mycelial growth of Cochliobolus heterostrophus was significantly hindered by A-ITC. However, the function of HK in regulating A-ITC sensitivity was not clear in C. heterostrophus, the causal agent of Southern corn leaf blight. RESULTS: In this study, the role of HKs was investigated in C. heterostrophus. Deletion of the HK coding gene ChNIK1 resulted in dramatically increased sensitivity of C. heterostrophus to A-ITC. In addition, ΔChnik1 mutant exhibited significantly decreased conidiation and increased sensitivity to NaCl, KCl, tebuconazole and azoxystrobin, but deletion of the other five HK genes did not affect the A-ITC sensitivity of C. heterostrophus. ChSLN1, ChNIK4, ChNIK8 and ChMAK2 are essential for conidiation and response to H2 O2 and sodium dodecyl sulfate. However, deletion of NIKs had on effect on significant virulence. CONCLUSION: Our findings demonstrate that the HKs play different roles in A-ITC sensitivity in C. heterostrophus. © 2023 Society of Chemical Industry.

Pantoea jilinensis D25 enhances tomato salt tolerance via altering antioxidant responses and soil microbial community structure.

As a major challenge to global food security, soil salinity is an important abiotic stress factor that seriously affects the crop growth and yield. In this study, the mechanism of salt resistance of Pantoea jilinensis D25 and its improving effect on salt tolerance of tomato were explored with salt resistance-related genes identified in strain D25 by genomic sequencing. The results showed that in comparison with the treatment of NaCl, strain D25 significantly increased the fresh weight, shoot length, root length, and chlorophyll content of tomato under salt stress by 46.7%, 20%, 42.4%, and 44.2%, respectively, with increased absorptions of various macronutrients and micronutrients and decreased accumulation of Na+. The activities of defense enzymes (peroxidase, catalase, superoxide dismutase, phenylalanine ammonia-lyase, and polyphenol oxidase) were enhanced, while the content of malondialdehyde was decreased. The results of quantitative real-time PCR analysis showed that the expressions of genes (SlSOS1, SlNHX1, SlHKT1.1, SlSOD1, SlAPX2, SlAOS, SlPin II, Solyc08g066270.1, Solyc03g083420.2 and SlGA20ox1) related to ion transporters, antioxidant machinery, key defense, serine/threonine protein kinase synthesis, and gibberellin (GA) signal protein were up-regulated and were the highest in the treatment of both NaCl and strain D25. The activities of enzymes (dehydrogenase, urease, invertase, and catalase activities) related to soil fertility were enhanced. The results of 16S rRNA sequencing showed that soil microbial diversity and the abundance of probiotics (e.g., Acidibacter, Limnobacter, and Romboutsia) were significantly increased. Our study provided strong experimental evidence to support the agricultural application of strain D25 in the promotion of growth in crops.

The Snf5-Hsf1 transcription module synergistically regulates stress responses and pathogenicity by maintaining ROS homeostasis in Sclerotinia sclerotiorum.

The SWI/SNF complex is guided to the promoters of designated genes by its co-operator to activate transcription in a timely and appropriate manner to govern development, pathogenesis, and stress responses in fungi. Nevertheless, knowledge of the complexes and their co-operator in phytopathogenic fungi is still fragmented. We demonstrate that the heat shock transcription factor SsHsf1 guides the SWI/SNF complex to promoters of heat shock protein (hsp) genes and antioxidant enzyme genes using biochemistry and pharmacology. This is accomplished through direct interaction with the complex subunit SsSnf5 under heat shock and oxidative stress. This results in the activation of their transcription and mediates histone displacement to maintain reactive oxygen species (ROS) homeostasis. Genetic results demonstrate that the transcription module formed by SsSnf5 and SsHsf1 is responsible for regulating morphogenesis, stress tolerance, and pathogenicity in Sclerotinia sclerotiorum, especially by directly activating the transcription of hsp genes and antioxidant enzyme genes counteracting plant-derived ROS. Furthermore, we show that stress-induced phosphorylation of SsSnf5 is necessary for the formation of the transcription module. This study establishes that the SWI/SNF complex and its co-operator cooperatively regulate the transcription of hsp genes and antioxidant enzyme genes to respond to host and environmental stress in the devastating phytopathogenic fungi.

Advances in Mechanism and Application of Molecular Breeding of Medicinal Mushrooms: A Review.

With the development of molecular biology and genomics technology, mushroom breeding methods have changed from single traditional breeding to molecular breeding. Compared with traditional breeding methods, molecular breeding has the advantages of short time and high efficiency. It breaks through the restrictive factors of conventional breeding and improves the accuracy of breeding. Molecular breeding technology is gradually applied to mushroom breeding. This paper summarizes the concept of molecular breeding and the application progress of various molecular breeding technologies in mushroom breeding, in order to provide reference for future research on mushroom breeding.

Mechanism of Liuwei Dihuang Pills in treating osteoporosis based on network pharmacology.

Osteoporosis is a prevalent age-related disease that poses a significant public health concern as the population continues to age. While current treatments have shown some therapeutic benefits, their long-term clinical efficacy is limited by a lack of stable curative effects and significant adverse effects. Traditional Chinese Medicine has gained attention due to its positive curative effects and fewer side effects. Liuwei Dihuang Pill has been found to enhance bone mineral density in patients with osteoporosis and rats, but the underlying mechanism is not yet clear. To shed more light on this problem, this study aims to explore the pharmacological mechanism of Liuwei Dihuang Pill in treating osteoporosis using network pharmacology and molecular docking. The findings indicate that Liuwei Dihuang Pills treat osteoporosis through various targets and channels. Specifically, it mainly involves TNF, IL17, and HIF-1 signaling pathways and helps regulate biological processes such as angiogenesis, apoptosis, hypoxia, and gene expression. Furthermore, molecular docking demonstrates excellent binding properties between the drug components and key targets. Therefore, this study offers a theoretical foundation for understanding the pharmacological mechanism and clinical application of Liuwei Dihuang Pills in treating osteoporosis more comprehensively.

Antifungal activity of Klebsiella grimontii DR11 against Fusarium oxysporum causing soybean root rot.

AIMS: Soybean root rot, caused by Fusarium oxysporum, leads to significant economic and financial losses to the soybean processing industry globally. In the study, we aimed to explore a biocontrol agent to combat F. oxysporum infection in soybean. METHODS AND RESULTS: From soybean rhizosphere soil, 48 strains were isolated. Among them, the strain DR11 exhibited the highest inhibition rate of 72.27%. Morphological, physiological, biochemical, and 16S rDNA identification revealed that the strain DR11 was Klebsiella grimontii DR11. Strain DR11 could inhibit the growth of F. oxysporum and spore formation and alter the mycelial morphology. At 5.0 × 106 CFU mL-1, pH 7, and 30°C, it exhibited the highest inhibitory rate (72.27%). Moreover, it could decrease the activity of cell-wall-degrading enzymes of F. oxysporum. Simultaneously, the activities of defense-related enzymes and content of malondialdehyde in soybean plants were increased after treatment with strain DR11. In addition, strain DR11 could form aggregates to form biofilm and adsorb on the surface of soybean roots. It inhibited F. oxysporum growth on soybean seedlings, with an inhibitory effect of 62.71%. CONCLUSION: Klebsiella grimontii DR11 had a strong inhibitory effect on F. oxysporum and could be used as a biocontrol agent to combat F. oxysporum infection in soybean.

Crucial Roles of the High-Osmolarity Glycerol Pathway in the Antifungal Activity of Isothiocyanates against Cochliobolus heterostrophus.

Isothiocyanates (ITCs) that are found in Brassicaceae exhibited obvious antifungal activity against Cochliobolus heterostrophus, which is the causal agent of southern corn leaf blight. However, the underlying antifungal mechanism of allyl-ITCs (A-ITCs) against C. heterostrophus remains largely unknown. Here, we used transcriptomic analysis to find that the high osmolarity pathway was upregulated significantly when treated with A-ITCs. To investigate the roles of the high osmolarity pathway in adaption to A-ITCs, we constructed Δssk2, Δpbs2, and Δhog1 mutant strains. Deletion of three genes (ChSSK2, ChPBS2, and ChHOG1) involved in the high osmolarity pathway resulted in significantly increased sensitivity of C. heterostrophus to ITCs. In addition, the phosphorylation level of ChHog1 was induced by A-ITC and was dependent on the presence of ChSsk2 and ChPbs2. Moreover, Δssk2, Δpbs2, and Δhog1 mutants exhibited a dramatically decreased virulence on maize leaves. Our findings demonstrated that the high osmolarity pathway played a positive role in ITC tolerance and virulence, which may provide novel insights into developing ITCs as a new fungicide against C. heterostrophus.

IL-1ß-mediated inflammatory responses in intervertebral disc degeneration: Mechanisms, signaling pathways, and therapeutic potential.

Intervertebral disc degeneration (IDD) has been widely recognized as the primary cause of low back pain and is one of the major chronic diseases imposing a severe socioeconomic burden worldwide. IDD is a degenerative process characterized by inflammatory responses, and its underlying pathological mechanisms remain complex. Genetic, developmental, biochemical, and biomechanical factors contribute to the development of IDD. There is a pressing need for an effective non-surgical treatment, mainly due to the lack of comprehensive understanding of the specific mechanisms involved and the effective therapeutic targets for IDD. Recently, interleukin (IL)-1ß has been recognized as an essential inflammatory factor and a key mediator of the inflammatory process in IDD. Current studies have found that IL-1ß is mainly involved in IDD by affecting the metabolism of the extracellular matrix and regulating cell death (RCD), such as apoptosis, pyroptosis, and ferroptosis (a new form of RCD). Although analysis of clinical samples from different laboratories confirmed how IL-1ß is induced in IDD, its specific signal transduction pathway, and the inflammatory role mediated in IDD remains unclear. This review describes the molecules and mechanisms involved in IL-1ß-mediated inflammatory responses, and their roles in resolving the inflammatory process in IDD. Understanding the signaling pathways involved in IL-1ß may lead to a new class of targets that promote remission for IDD patients. This review aims to provide a framework for the treatment of IDD by analyzing the signaling mechanism and function related to IL-1ß, especially in terms of inflammation, matrix metabolism, and cell death regulation.

From fetal tendon regeneration to adult therapeutic modalities: TGF-ß3 in scarless healing.

Tendon injuries are common disorders that can significantly impact people's lives. Unfortunately, the limited regenerative ability of tendons results in tissue healing in a scar-mediated manner. The current therapeutic strategies fail to fully recover the functions of the injured tendons, and as such, the conception of 'scarless healing' has gained prominent attention in the field of regenerative medicine. Interestingly, injured fetal tendons possess the capability to heal through regeneration, which builds an ideal blueprint for adult tendon regeneration. Studies have shown that fetal biochemical cues have the potential to improve adult tendon healing. Here we review the biological factors that contribute to fetal tendon regeneration and how manipulation of these biochemical cues in the adult tendon healing process could achieve regeneration.

We reviewed the biological factors that contribute to fetal tendon regeneration and how manipulation of these biochemical cues in the adult tendon healing process could achieve regeneration. The results showed that inflammation and TGF-ß level are the main elements involved in fetal tendon regeneration. Experimental manipulation of these biochemical cues in the adult tendon healing process demonstrated that although the blockade of TGF-ß1, TGF-ß2 and inflammation reduced scar tissue in adult tendon healing, this inhibition also destroyed the mechanical properties of the tendons. An effective alternative is regulating the specific downstream profibrotic effectors of both TGF-ß1 and inflammation, which is preferable to those that completely inhibit these factors. Finally, TGF-ß3 is a master regulator allowing a shift from adult scar healing to scarless healing, and the administration of TGF-ß3 is a viable strategy to promote adult regenerative healing. In terms of mechanisms, TGF-ß3 can activate Smad7 and inhibit the JNK/c-Jun signaling pathway to promote tendon regenerative healing.

Strain Field Characteristics of Coal-like Material under CO2 Fracturing.

Considering the problems related to hydrofracturing, CO2 fracturing is a more effective way to enhance the permeability of coal seams. Conventional research has focused on the dynamic behavior of coal and rock under CO2 fracturing. There is a lack of quantitative descriptions of the fracturing stages. Therefore, CO2 fracturing experiments were carried out with coal-like material at several pressures. The maximum principal strain field was obtained by analyzing the surface images of the specimens. Nine characteristic parameters of typical grayscale and texture images were calculated from the strain field. The correlation between the maximum principal strain and the characteristic parameters was examined using gray relational analysis, and the effects of pressure on those parameters were investigated. We obtained some interesting results, for example, the standard deviation increased from 50 to 100, and kurtosis decreased from -1.0 to -1.5 with the fracturing pressure. The thresholds of the characteristic function F at different fracturing stages were determined, too. This research provides a basis for designing the drilling hole location and determining the millisecond delay of multihole fracturing, as well as for automatically dividing the fracturing stages with artificial intelligence based on the maximum principal strain field.

Preoperative Discrimination of CDKN2A/B Homozygous Deletion Status in Isocitrate Dehydrogenase-Mutant Astrocytoma: A Deep Learning-Based Radiomics Model Using MRI.

BACKGROUND: Cyclin-dependent kinase inhibitor 2A/B (CDKN2A/B) homozygous deletion has been verified as an independent and critical biomarker of negative prognosis and short survival in isocitrate dehydrogenase (IDH)-mutant astrocytoma. Therefore, noninvasive and accurate discrimination of CDKN2A/B homozygous deletion status is essential for the clinical management of IDH-mutant astrocytoma patients. PURPOSE: To develop a noninvasive, robust preoperative model based on MR image features for discriminating CDKN2A/B homozygous deletion status of IDH-mutant astrocytoma. STUDY TYPE: Retrospective. POPULATION: Two hundred fifty-one patients: 107 patients with CDKN2A/B homozygous deletion and 144 patients without CDKN2A/B homozygous deletion. FIELD STRENGTH/SEQUENCE: 3.0 T/1.5 T: Contrast-enhanced T1-weighted spin-echo inversion recovery sequence (CE-T1WI) and T2-weighted fluid-attenuation spin-echo inversion recovery sequence (T2FLAIR). ASSESSMENT: A total of 1106 radiomics and 1000 deep learning features extracted from CE-T1WI and T2FLAIR were used to develop models to discriminate the CDKN2A/B homozygous deletion status. Radiomics models, deep learning-based radiomics (DLR) models and the final integrated model combining radiomics features with deep learning features were developed and compared their preoperative discrimination performance. STATISTICAL TESTING: Pearson chi-square test and Mann Whitney U test were used for assessing the statistical differences in patients' clinical characteristics. The Delong test compared the statistical differences of receiver operating characteristic (ROC) curves and area under the curve (AUC) of different models. The significance threshold is P < 0.05. RESULTS: The final combined model (training AUC = 0.966; validation AUC = 0.935; test group: AUC = 0.943) outperformed the optimal models based on only radiomics or DLR features (training: AUC = 0.916 and 0.952; validation: AUC = 0.886 and 0.912; test group: AUC = 0.862 and 0.902). DATA CONCLUSION: Whether based on a single sequence or a combination of two sequences, radiomics and DLR models have achieved promising performance in assessing CDKN2A/B homozygous deletion status. However, the final model combining both deep learning and radiomics features from CE-T1WI and T2FLAIR outperformed the optimal radiomics or DLR model. EVIDENCE LEVEL: 4 TECHNICAL EFFICACY: Stage 2.

CDK12 loss inhibits cell proliferation by regulating TBK1 in non-small cell lung cancer cells.

Lung cancer is one of the most common malignant tumors and has a poor prognosis and a low survival rate. Traditional treatments, such as radiotherapy and chemotherapy, still face some challenges because of high drug resistance and toxicity. Therefore, it is necessary to discover a new kind of targeted drug with low toxicity and high efficiency. CDK12 is a cell cycle-dependent kinase whose main function is to activate RNA polymerase II (RNAPII) and promote the transcriptional extension of RNA. However, the role and molecular mechanism of CDK12 in lung cancer are still unclear. In this study, the mutation and RNA-Seq data of CDK12 in lung adenocarcinoma and squamous cell carcinoma were downloaded from The Cancer Genome Atlas (TCGA) database and analyzed with the custom scripts. Cell proliferation was evaluated by Cell Counting Kit-8 (CCK-8) and cell colony formation assays. A subcutaneous tumor experiment in nude mice was used to examine the effects of CDK12 knockdown on the in vivo tumor growth of NSCLC cells. The cell cycle distribution and the apoptosis rate of lung cancer cells were assessed by flow cytometry. Regulation of TANK-binding kinase 1 (TBK1) by CDK12 was evaluated by quantitative PCR, immunoprecipitation and Western blot analysis. In this study we have analyzed the mutation and expression data of The Cancer Genome Atlas (TCGA) database and found that CDK12 is highly expressed in lung cancer tissues. Clinical correlation analysis showed that high expression of CDK12 in NSCLC reduces patient survival, but its high expression is only related to early tumor progression and has no significant correlation with late tumor progression and metastasis. Furthermore, we present evidence that CDK12 depletion in lung cancer cell lines not only leads to the inhibition of cell growth and induces apoptosis but also inhibits tumor growth of NSCLC cells in vivo. CDK12 positively regulates the expression of the oncogene TBK1 in lung cancer cells. These results revealed that CDK12 affects the progression of non-small cell lung cancer through positive regulation of TBK1 expression, suggesting that CDK12 might be a potential molecular target for the treatment of non-small cell lung cancer.