Discrete RNA-DNA hybrid cleavage by the EXD2 exonuclease pinpoints two rate-limiting steps.

EXD2 is a recently identified exonuclease that cleaves RNA and DNA in double-stranded (ds) forms. It thus serves as a model system for investigating the similarities and discrepancies between exoribonuclease and exodeoxyribonuclease activities and for understanding the nucleic acid (NA) unwinding-degradation coordination of an exonuclease. Here, using a single-molecule fluorescence resonance energy transfer (smFRET) approach, we show that despite stable binding to both substrates, EXD2 barely cleaves dsDNA and yet displays both exoribonuclease and exodeoxyribonuclease activities toward RNA-DNA hybrids with a cleavage preference for RNA. Unexpectedly, EXD2-mediated hybrid cleavage proceeds in a discrete stepwise pattern, wherein a sudden 4-bp duplex unwinding increment and the subsequent dwell constitute a complete hydrolysis cycle. The relatively weak exodeoxyribonuclease activity of EXD2 partially originates from frequent hybrid rewinding. Importantly, kinetic analysis and comparison of the dwell times under varied conditions reveal two rate-limiting steps of hybrid unwinding and nucleotide excision. Overall, our findings help better understand the cellular functions of EXD2, and the cyclic coupling between duplex unwinding and exonucleolytic degradation may be generalizable to other exonucleases.

Phase-separated ParB enforces diverse DNA compaction modes and stabilizes the parS-centered partition complex.

The tripartite ParABS system mediates chromosome segregation in the majority of bacterial species. Typically, DNA-bound ParB proteins around the parS sites condense the chromosomal DNA into a higher-order multimeric nucleoprotein complex for the ParA-driven partition. Despite extensive studies, the molecular mechanism underlying the dynamic assembly of the partition complex remains unclear. Herein, we demonstrate that Bacillus subtilis ParB (Spo0J), through the multimerization of its N-terminal domain, forms phase-separated condensates along a single DNA molecule, leading to the concurrent organization of DNA into a compact structure. Specifically, in addition to the co-condensation of ParB dimers with DNA, the engagement of well-established ParB condensates with DNA allows for the compression of adjacent DNA and the looping of distant DNA. Notably, the presence of CTP promotes the formation of condensates by a low amount of ParB at parS sites, triggering two-step DNA condensation. Remarkably, parS-centered ParB-DNA co-condensate constitutes a robust nucleoprotein architecture capable of withstanding disruptive forces of tens of piconewton. Overall, our findings unveil diverse modes of DNA compaction enabled by phase-separated ParB and offer new insights into the dynamic assembly and maintenance of the bacterial partition complex.

Application of TiLOOP bra in implant-based breast reconstruction is associated with decreased complication risk compared with other meshes: A systematic review and meta-analysis.

BACKGROUND: TiLOOP bra has been used for over 15 years, however, evidence regarding its safety in implant-based breast reconstruction (IBBR) for patients with breast cancer after mastectomy is still limited. We performed this meta-analysis to evaluate its risks and benefits in IBBR comparing with other meshes. METHODS: Electronic databases were searched to identify relevant studies comparing postoperative complications between TiLOOP bra and other reconstruction techniques in IBBR with or without meshes. We also compared patient satisfaction in physical well-being between two groups. Risk ratios (RRs) and mean differences with 95% confidence interval (CI) were calculated. RESULTS: Seven studies representing 1203 cases were analyzed. Compared with other meshes, the use of TiLOOP bra significantly reduced the risk of infection (RR = 0.53, 95% CI, 0.32-0.86), seroma (RR = 0.21, 95% CI, 0.07-0.61), red breast syndrome (RR = 0.10, 95% CI, 0.02-0.45), and capsular contracture (RR = 0.20, 95% CI, 0.05-0.75). Patient satisfaction in physical well-being was comparable between two groups. CONCLUSIONS: TiLOOP bra in IBBR has a favored safety profile over other meshes, which significantly reduced postoperative complication risk and did not affect patient satisfaction. Although prospective well-designed controlled studies are still warranted, TiLOOP bra is safe and reliable at present.

Stochastically multimerized ParB orchestrates DNA assembly as unveiled by single-molecule analysis.

The tripartite ParABS system mediates chromosome segregation in a wide range of bacteria. Dimeric ParB was proposed to nucleate on parS sites and spread to neighboring DNA. However, how properly distributed ParB dimers further compact chromosomal DNA into a higher-order nucleoprotein complex for partitioning remains poorly understood. Here, using a single-molecule approach, we show that tens of Bacillus subtilis ParB (Spo0J) proteins can stochastically multimerize on and stably bind to nonspecific DNA. The introduction of CTP promotes the formation and diffusion of the multimeric ParB along DNA, offering an opportunity for ParB proteins to further forgather and cluster. Intriguingly, ParB multimers can recognize parS motifs and are more inclined to remain immobile on them. Importantly, the ParB multimer features distinct capabilities of not only bridging two independent DNA molecules but also mediating their transportation, both of which are enhanced by the presence of either CTP or parS in the DNA. These findings shed new light on ParB dynamics in self-multimerization and DNA organization and help to better comprehend the assembly of the ParB-DNA partition complex.

Higher Dietary Inflammatory Index and Systemic Immune-Inflammation Index Score are Associated With Higher Risk of Chronic Kidney Disease: Analysis of the National Health and Nutrition Examination Survey From 1999 to 2018.

OBJECTIVE: Chronic kidney disease (CKD) is characterized by a gradual decline in kidney function over time. The role of dietary inflammatory index (DII) and systemic immune-inflammation index (SII) in individuals with CKD remains uncertain. We aimed to explore the potential correlation between DII and SII with the prevalence of CKD in adult Americans. METHODS: This cross-sectional study used data from the National Health and Nutrition Examination Study between 1999 and 2018. The DII was calculated based on the 24-hour dietary history interview, while the SII was calculated as the product of platelet count multiplied by neutrophil count and divided by lymphocyte count. CKD was diagnosed based on impaired glomerular filtration rate (<60 mL/min per 1.73 m2) or urinary albumin-creatinine ratio ≥30 mg/g. Multivariable logistic regression analyses and subgroup analyses were performed to examine the association between DII/SII and CKD. RESULTS: In total, this study included 40,388 participants, of whom 7443 (18.4%) had CKD. The prevalence of CKD changed from 14.84% (95% confidence interval (CI): 13.20-16.48%) in 1999-2000 to 12.76% (95% CI: 11.10-14.43%) in 2017-2018. According to adjusted multivariate logistic regression models, individuals with higher DII scores had a higher likelihood of having CKD (odds ratio = 1.24; 95% CI: 1.12-1.37). Similarly, higher SII scores were associated with a higher risk of CKD (odds ratio = 1.37; 95% CI: 1.25-1.50). Subgroup analyses further demonstrated relatively stronger associations between DII/SII and CKD among individuals with other factors such as sex, age, body mass index, smoking status, drinking status, hypertension, and diabetes. CONCLUSIONS: The DII and SII scores were significantly positively associated with higher risks of CKD. Anti-inflammatory diet might have the potential to prevent CKD. The SII may serve as a cost-effective and straightforward approach for detecting CKD. Further prospective longitudinal studies are needed to verify the causality.

Rh(III)-Catalyzed C-H Annulation of N-Nitrosoanilines with Iodonium Ylides for the Synthesis of N-Alkyl Indoles.

A novel protocol for synthesizing N-alkyl indoles from readily available N-nitrosoanilines and iodonium ylides through the rhodium(III)-catalyzed C-H bond activation/intramolecular cyclization reaction has been described. This strategy employs nitroso as a traceless directing group. The transformation features powerful reactivity, tolerates various functional groups, and proceeds with moderate yields under mild reaction conditions, providing a straightforward approach to access structurally diverse and valuable N-alkyl indole derivatives.

Structural evolution and mechanical stabilities of head-to-side nanowelding of Cu-Ag bimetallic nanowires via atomistic simulations.

Nanowelding, self-healing and mechanical stabilities of conductive networks of Cu-Ag core-shell nanowires are of vital importance for their extensive applications. In this study, atomistic simulations are used to reveal the head-to-side cold welding behavior, ranging from the welding mechanism, mechanical stabilities of the obtained junction and effects of various conditions. The results show that head-to-side cold welding of Cu-Ag bimetallic nanowires can be excellently completed via atomic interaction and diffusion of atoms. Initial deformation in the junction induced in the welding process and welding temperature are proven to exert a significant influence on the mechanical stabilities of the obtained junction. Three different deformation mechanisms are proposed due to various motivations of dislocations. During the uniaxial tensile test of the junction, the plastic deformation map of initial deformation and welding temperature are expounded in detail. It is revealed that for all the involved welding temperatures explored in our study, the highest tensile strength always belongs to the T-junction with no initial deformation. Otherwise, the intersection will become a serious obstacle to a further process of plastic deformation and lead to abnormally larger elongation and lower strength. These findings are expected to provide an in-depth understanding of the deformation mechanism of bimetallic nanowires and provide valuable theoretical guidance for engineering applications.

Dynamics of Staphylococcus aureus Cas9 in DNA target Association and Dissociation.

Staphylococcus aureus Cas9 (SaCas9) is an RNA-guided endonuclease that targets complementary DNA adjacent to a protospacer adjacent motif (PAM) for cleavage. Its small size facilitates in vivo delivery for genome editing in various organisms. Herein, using single-molecule and ensemble approaches, we systemically study the mechanism of SaCas9 underlying its interplay with DNA. We find that the DNA binding and cleavage of SaCas9 require complementarities of 6- and 18-bp of PAM-proximal DNA with guide RNA, respectively. These activities are mediated by two steady interactions among the ternary complex, one of which is located approximately 6 bp from the PAM and beyond the apparent footprint of SaCas9 on DNA. Notably, the other interaction within the protospacer is significantly strong and thus poses DNA-bound SaCas9 a persistent block to DNA-tracking motors. Intriguingly, after cleavage, SaCas9 autonomously releases the PAM-distal DNA while retaining binding to the PAM. This partial DNA release immediately abolishes its strong interaction with the protospacer DNA and consequently promotes its subsequent dissociation from the PAM. Overall, these data provide a dynamic understanding of SaCas9 and instruct its effective applications.

3-Aminophenylboronic acid-functionalized molybdenum disulfide quantum dots for fluorescent determination of hypochlorite.

A simple method is reported for hypochlorite determination based on fluorescence 3-aminophenylboronic acid-functionalized molybdenum disulfide quantum dots (B-MoS2 QDs). B-MoS2 QDs with strong fluorescence at 380 nm have been successfully synthesized by the amidation reaction between APBA and hydrothermal MoS2 QDs. Hypochlorite sensing was proposed utilizing the fluorescent quenching effect of 3,3',5,5'-tetramethylbenzidine dihydrochloride (TMB) on B-MoS2 QDs and the fast redox reaction between hypochlorite and TMB. The fluorescent quenching effect of TMB to B-MoS2 QDs was proved to be caused by static dynamic quenching and inner filter effect. A good linear relationship was obtained in the hypochlorite concentration range from 1 to 20 µM, and the limit of detection (LOD) was 36.8 nM. The proposed fluorescent detection assay was simple and fast, taking only 5 min at room temperature. Satisfactory results were obtained in the standard spike recovery tests on tap water and milk samples, which indicate high potential in constructing fluorescent bio-detection assays.

Congenital absence of left atrial appendage combined with type A Wolff-Parkinson-White syndrome diagnosed by multimodal imaging.

The absence of left atrial appendage (LAA) is relatively rare, especially with type A Wolff-Parkinson-White syndrome. Secondly, we diagnosed it by multimodal imaging including two-dimensional (2D) and three-dimensional (3D) transesophageal echocardiography (TEE), CT, electrophysiological examination, and 3D electro anatomical mapping system, which is more comprehensive.

Tumor-Derived Small Extracellular Vesicles Involved in Breast Cancer Progression and Drug Resistance.

Breast cancer is one of the most serious and terrifying threats to the health of women. Recent studies have demonstrated that interaction among cancer cells themselves and those with other cells, including immune cells, in a tumor microenvironment potentially and intrinsically regulate and determine cancer progression and metastasis. Small extracellular vesicles (sEVs), a type of lipid-bilayer particles derived from cells, with a size of less than 200 nm, are recognized as one form of important mediators in cell-to-cell communication. sEVs can transport a variety of bioactive substances, including proteins, RNAs, and lipids. Accumulating evidence has revealed that sEVs play a crucial role in cancer development and progression, with a significant impact on proliferation, invasion, and metastasis. In addition, sEVs systematically coordinate physiological and pathological processes, such as coagulation, vascular leakage, and stromal cell reprogramming, to bring about premetastatic niche formation and to determine metastatic organ tropism. There are a variety of oncogenic factors in tumor-derived sEVs that mediate cellular communication between local stromal cells and distal microenvironment, both of which are important in cancer progression and metastasis. Tumor-derived sEVs contain substances that are similar to parental tumor cells, and as such, sEVs could be biomarkers in cancer progression and potential therapeutic targets, particularly for predicting and preventing future metastatic development. Here, we review the mechanisms underlying the regulation by tumor-derived sEVs on cancer development and progression, including proliferation, metastasis, drug resistance, and immunosuppression, which coordinately shape the pro-metastatic microenvironment. In addition, we describe the application of sEVs to the development of cancer biomarkers and potential therapeutic modalities and discuss how they can be engineered and translated into clinical practice.

Bloom Syndrome Helicase Compresses Single-Stranded DNA into Phase-Separated Condensates.

Bloom syndrome protein (BLM) is a conserved RecQ family helicase involved in the maintenance of genome stability. BLM has been widely recognized as a genome "caretaker" that processes structured DNA. In contrast, our knowledge of how BLM behaves on single-stranded (ss) DNA is still limited. Here, we demonstrate that BLM possesses the intrinsic ability for phase separation and can co-phase separate with ssDNA to form dynamically arrested protein/ssDNA co-condensates. The introduction of ATP potentiates the capability of BLM to condense on ssDNA, which further promotes the compression of ssDNA against a resistive force of up to 60 piconewtons. Moreover, BLM is also capable of condensing replication protein A (RPA)- or RAD51-coated ssDNA, before which it generates naked ssDNA by dismantling these ssDNA-binding proteins. Overall, our findings identify an unexpected characteristic of a DNA helicase and provide a new angle of protein/ssDNA co-condensation for understanding the genomic instability caused by BLM overexpression under diseased conditions.

The regulatory effect of acetylation of HMGN2 and H3K27 on pyocyanin-induced autophagy in macrophages by affecting Ulk1 transcription.

Pyocyanin (PYO) is a major virulence factor secreted by Pseudomonas aeruginosa, and autophagy is a crucial homeostatic mechanism for the interaction between the pathogens and the host. It remains unknown whether PYO leads to autophagy in macrophages by regulating histone acetylation. The high mobility group nucleosomal binding domain 2 (HMGN2) has been reported to regulate the PYO-induced autophagy and oxidative stress in the epithelial cells; however, the underlying molecular mechanism has not been fully elucidated. In this study, PYO was found to induce autophagy in macrophages, and the mechanism might be correlated with the up-regulation of HMGN2 acetylation (HMGN2ac) and the down-regulation of H3K27 acetylation (H3K27ac) by modulation of the activities of acetyltransferases and deacetylases. Moreover, we further demonstrated that the up-regulated HMGN2ac enhances its recruitment to the Ulk1 promoter, while the down-regulation of H3K27ac reduces its recruitment to the Ulk1 promoter, thereby promoting or inhibiting the transcription of Ulk1. In conclusion, HMGN2ac and H3K27ac play regulatory roles in the PYO-induced autophagy in macrophages.

Dynamic Changes in Plasma Urotensin II and Its Correlation With Plaque Stability.

ABSTRACT: Urotensin II (UII) is involved in the formation of atherosclerosis, but its role in the stability of atherosclerotic plaques is unknown. The purpose of this study was to observe the dynamic changes in plasma UII and analyze its relationship to the stability of atherosclerotic plaques. One hundred thirty-five consecutive patients with acute coronary syndrome (ACS) were enrolled. The plasma UII levels were measured immediately after admission and during three-month follow-up. A vulnerable plaque model was established using local transfection of a recombinant P53 adenovirus into plaques in rabbits fed with a high-cholesterol diet and subjected to balloon arterial injury. The levels of plasma UII were measured weekly. The changes in plasma UII during the formation of atherosclerotic plaques and before and after plaque transfection were observed. The morphology of the plaques and the expression, distribution, and quantitative expression of UII in the plaques also were observed. Our results showed that the levels of plasma UII in patients with ACS at admission were lower than levels observed at the three-month follow-up. UII dynamic changes and its correlation with plaque stabilities were further verified in rabbits with atherosclerotic vulnerable plaques. The UII levels in rabbits were significantly decreased immediately after the P53 gene transfection, which led to plaque instability and rupture. These results suggested that UII expression was down-regulated in ACS, which may be related to its ability to modulate mechanisms involved in plaque stability and instability.

Glutaredoxin 1 up-regulates deglutathionylation of α4 integrin and thereby restricts neutrophil mobilization from bone marrow.

α4 integrin plays a crucial role in retention and release of neutrophils from bone marrow. Although α4 integrin is known to be a potential target of reactive oxygen species (ROS)-induced cysteine glutathionylation, the physiological significance and underlying regulatory mechanism of this event remain elusive. Here, using in vitro and in vivo biochemical and cell biology approaches, we show that physiological ROS-induced glutathionylation of α4 integrin in neutrophils increases the binding of neutrophil-associated α4 integrin to vascular cell adhesion molecule 1 (VCAM-1) on human endothelial cells. This enhanced binding was reversed by extracellular glutaredoxin 1 (Grx1), a thiol disulfide oxidoreductase promoting protein deglutathionylation. Furthermore, in a murine inflammation model, Grx1 disruption dramatically elevated α4 glutathionylation and subsequently enhanced neutrophil egress from the bone marrow. Corroborating this observation, intravenous injection of recombinant Grx1 into mice inhibited α4 glutathionylation and thereby suppressed inflammation-induced neutrophil mobilization from the bone marrow. Taken together, our results establish ROS-elicited glutathionylation and its modulation by Grx1 as pivotal regulatory mechanisms controlling α4 integrin affinity and neutrophil mobilization from the bone marrow under physiological conditions.

Relationship between GDF15 level and bone metabolism in postmenopausal Chinese women.

Background: Growth differentiation factor 15 (GDF15), is a newly identified member of the transforming growth factor-beta (TGF-ß) family. It circulates as a 24.5-kDa homodimer. However, the function of GDF15 in bone metabolism remains unclear. In this study, we investigated the function of GDF15 in postmenopausal Chinese women.Methods: We measured serum GDF15 levels, bone mineral density (BMD), and bone turnover markers in 201 postmenopausal Chinese women ranging in age from 47 to 80 years.Results: The concentration of serum GDF15 increased with age. Growth differentiation factor 15 levels displayed a negative correlation with lumbar spine, femoral neck, and total hip BMD. After adjusting for age, this association still existed and was significant. We identified age, GDF15, body mass index (BMI), and estradiol to be associated with BMD. Furthermore, we found that GDF15 levels had a significant negative relationship with bone alkaline phosphatase (BAP) levels; this relationship remained significant after adjustment. However, there was no significant correlation between levels of GDF15 and N-terminal telopeptide of type I collagen (NTX).Conclusions: For postmenopausal Chinese women, GDF15 is a negative predictor of BMD and has a negative correlation with bone formation biomarker BAP. In other words, GDF15 exerts negative regulation on bone mass by inhibiting bone formation.

Scopulariopsis/Microascus isolation in lung transplant recipients: A report of three cases and a review of the literature.

The current knowledge of invasive Scopulariopsis/Microascus infection in lung transplantation has been derived from only four case reports. Although these fungi are uncommon compared with Aspergillus, they are highly resistant to the current antifungal agents, and the mortality is extremely high. To explore the risk factors, clinical manifestations, notable diagnostic characteristics and outcomes of positive Scopulariopsis/Microascus isolation in lung transplantation patients. We included all cases with positive Scopulariopsis/Microascus isolation from lower respiratory tracts or bronchial mucosa biopsies in our lung transplantation centre. Proven cases from the literature were added. Positive isolation occurred in 2% (3/157) in our centre. Four cases from the literature were added. The mortality could be considered as high as 80%, once the two cases of colonisation were excluded. The average interval between transplantation and positive isolation was 106 (19-131) days. A total of 57.1% of patients had experienced a combination of infection with Aspergillus or other fungi as well as long-term azole antifungal agent treatment before the positive isolation, which may be possible risk factors. The combination of micafungin, posaconazole and terbinafine may be an effective treatment. The peak time of positive isolation was consistent with that of some opportunistic pathogens, and the possible risk factors were the infection of other fungi as well as prior long-term azole antifungal administration. In addition to its high mortality, Scopulariopsis/Microascus was also highly resistant to common antifungal agents and the combination of two or three drugs for therapy was recommended.

Effect of citral on mouse hepatic cytochrome P450 enzymes.

CONTEXT: Citral is used as a potential natural treatment for various infectious diseases. OBJECTIVE: To examine the effect of citral on the mRNA expression and activities of cytochrome P450 (CYP450) enzymes and establish the relationship between citral-induced liver injury and oxidative stress. MATERIALS AND METHODS: ICR mice were randomly divided into citral (20, 200, and 2000 mg/kglow), Tween-80, and control groups (0.9% saline), 10 mice in each group. The citral-treated groups were intragastrically administered citral for 3 d, control groups treated with 0.5% Tween-80 and 0.9% saline in the same way. Liver injury and CYP450 enzymes were analyzed by analyzing the histopathological changes and the changes of related enzymes. RESULTS: Citral treatment (2000 mg/kg) for 3 d increased serum glutamic pyruvic transaminase and glutamic oxaloacetic transaminase levels, as well as glutathione, gydroxyl radicals, malonaldehyde and total superoxide dismutase contents, but decreased the content of total antioxidant capacity. In doses of 20 and 200 mg/kg groups mice, the contents of NO were decreased significantly and other changes were similar to the 2000 mg/kg group mice, but the liver damage was most severe in the 2000 mg/kg group. Citral induced the mRNA expression and activities of CYP450 1A2, 2D22, and 2E1 in the liver of mice at doses of 20 and 200 mg/kg. There were no changes in testing indexes in Tween-80 treated group mice. Due to its toxic effects, the CYP induction effect of citral negatively correlated with its dose. Although the mRNA expression of CYP450 3A11 was induced by citral, its activity was not affected by low and moderate doses of citral. CYP450 3A11 activity was significantly decreased by high-dose citral. CONCLUSIONS: Citral is hepatotoxic and induced oxidative stress in higher dose, which has a negative effect on CYP450 enzymes. These data suggest caution needs to be taken in order to avoid citral-drug interactions in human beings.

Microencapsulation of low-passage poorly-differentiated human mucoepidermoid carcinoma cells by alginate microcapsules: in vitro profiling of angiogenesis-related molecules.

BACKGROUND: Human mucoepidermoid carcinoma (MEC) is regarded as the most common primary salivary malignancy. High-grade MEC has a high risk of recurrence and poor prognosis. Tumor angiogenesis, induced by poorly differentiated cancer cells of high-grade MEC, contributes to tumor growth and metastasis. Therefore, elucidating molecular mechanisms underlying the pro-angiogenic ability of poorly differentiated MEC cells is critical for the understanding of high-grade MEC progression. It is well known that three-dimensional (3D) cell culture, in contrast with conventional two-dimensional (2D) culture, provides a better approach to in vitro recapitulation of in vivo characteristics of cancer cells and their surrounding microenvironment. The purpose of this study was to model a 3D environment for in vitro gene expression profiling of key molecules in poorly differentiated MEC cells for cancer neovascularization and compared them with traditional 2D cell culture. METHODS: Low-passage poorly differentiated MEC cells, derived from human patient samples of high-grade MEC, were microencapsulated in sodium alginate gel microcapsules (3D culture) and compared with cells grown in 2D culture. Cancer cell proliferation was determined by MTT assays for 1 week, and gene expression of VEGF-A, bFGF and TSP-1 was analyzed by western blotting or ELISA. The hypoxic environment in 3D versus 2D culture were assessed by western blotting or immunofluorescence for HIF1α, and the effect of hypoxia on VEGF-A gene expression in 3D cultured cancer cells was assessed by western blotting with the use of the HIF1α inhibitor, 2-methoxyestradiol (2-MeOE2). RESULTS: When encapsulated in alginate gel microcapsules, low-passage poorly differentiated human MEC cells grew in blocks and demonstrated stronger and relatively unlimited proliferation activities. Moreover, significant differences were found in gene expression, with 3D-grown cancer cells a significant increment of VEGF-A and bFGF and a drastic reduction of TSP-1. Consistently, 3D-grown cancer cells secreted significantly more VEGF-A than 2D culture cancer cells. Furthermore, 3D-grown cancer cells showed significantly higher expression of HIF1α, a molecular indicator of hypoxia; the increased expression of VEGF-A in 3D cultured cancer cells was shown to be dependent on the HIF1α activities. CONCLUSIONS: The present work shows the effects of 3D culture model by alginate microencapsulation on the proangiogenic potentials of low-passage poorly differentiated human MEC cells. Cancer cells in this 3D system demonstrate significant intensification of key molecular processes for tumor angiogenesis. This is due to a better modeling of the hypoxic tumor microenvironment during 3D culture.

Specific imaging characteristic of solitary necrotic nodule of the liver: Marked peripheral rim-like enhancement with internal hypointensity on longer delayed MRI.

OBJECTIVES: To obtain specific imaging findings of solitary necrotic nodule of the liver (SNNL) using longer delayed contrast-enhanced MRI and compare them with those of three mimic hepatic diseases. METHODS: Sixteen patients with SNNL underwent plain and contrast-enhanced triphasic CT and multiphasic MRI with delayed time prolonged to 2 h after contrast bolus injection. Twenty-three patients with mimic lesions including seven with eight HCCs, five with five iCCs and 11 with metastatic lesions served as the control group. Those patients also received plain and multiphasic contrast-enhanced MRI. Imaging features of lesions such as peripheral wash-out time were evaluated. RESULTS: Among the 16 SNNLs, with a prolonged delayed MRI time, the enhancement degree of tumour periphery increased gradually. When it was up to 1 h, all lesions represented moderate/marked peripheral enhancement with internal hypointensity. However, the peripheral wash-out in seven HCCs (87.5%) and all metastatic lesions except three appeared at 10 or 15 min, one iCC (20%) at 30 min and the other lesions at 1 h. CONCLUSIONS: Longer MRI with a delayed time of 1-2 h may be useful in diagnosis SNNL, revealing the specific imaging characteristic of SNNL as pronounced peripheral enhancement with internal hypointensity. KEY POINTS: • Longer delayed MRI plays an important role in the diagnosis of SNNL. • Characteristic imaging feature of SNNL is pronounced peripheral enhancement with internal hypointensity. • Periphery wash-out time can differentiate SNNL from mimic diseases. • Imaging findings of SNNL on routine CT and MRI are unspecific.