Proliferative State Research Articles

High correlation between Ki-67 expression and a novel perfusion MRI biomarker diffusion-derived vessel density (DDVD) in endometrial carcinoma.

This study aimed to investigate the feasibility of diffusion-derived vessel density (DDVD) in characterizing tumor microvasculature in endometrial carcinoma (EC), and to explore the correlations with Ki-67 proliferation status and histological type based on DDVD values. There were in total 81 EC patients. There were 64 cases of non-aggressive histological type, and 17 cases of aggressive histological type. Ki-67 labeling index was low (<50%) in 35 cases and high (≥50%) in 46 cases. DDVD(b0b20) is calculated according to: DDVD(b0b20)=Sb0/ROIarea0 - Sb20/ROIarea20, where Sb0 and Sb20 refer to the tissue signal when b is 0 or 20s/mm2. Intraclass correlation coefficient (ICC); two-tailed independent samples t-test and Mann-Whitney U test, and Receiver operating characteristic area under the curve (AUC) were applied for statistical analysis. Endometrial carcinoma showed lower DDVD(b0b20) values (34.9±21.2, au/pixel) compared with myometrium (65.3±37.4, P<0.001). Tumors with Ki-67 high-proliferation or aggressive histological type had higher DDVD values than those with Ki-67 low-proliferation (44.17 (median) vs. 16.08, P<0.001]] or non-aggressive histological type (47.92 vs. 30.77, P=0.002). DDVD(b0b20) ROC curve analysis shows AUC of 0.842 for distinguishing between Ki-67 low- and high-expression, and AUC of 0.771 for distinguishing between non-aggressive and aggressive histological types. DDVD(b0b20)>32.9 and DDVD(b0b20)>50.1 provided a specificity of 85% for identifying Ki67 high expression (sensitivity 78.3%) and histological aggressive type (sensitivity 47.1%), respectively. DDVD can act as an imaging marker reflecting Ki-67 proliferation and histological aggressiveness of EC, thus helping pretreatment risk assessment in EC.

The mitochondrial and cytoplasmic superoxide anion imbalance trigger the expression of certain cellular aging markers in HaCaT keratinocytes.

In cells, the term "cellular aging" represents a collection of biological changes that can precede the proliferative senescence states. Cells more resistant to proliferative senescence, such as the ones found in the basal layer of the epidermis, may also exhibit these aging patterns. Therefore, cellular aging events could be induced by endogenous signals named here as cellular aging triggers (CATs) components. The superoxide anion (O2⁻) could be a prime candidate for a CATs, as it is continuously produced by eukaryotic cells. To test this hypothesis, mitochondrial and cytoplasmic O2⁻ imbalances were induced in HaCaT keratinocytes using rotenone (ROT, 30µM), which inhibits mitochondrial complex I and paraquat (PQT, 30µM), which increases O2⁻ levels via redox cycling. ROT and PQT reduced cellular proliferation rate and elevated β-Galactosidase and transforming growth factor beta (TGF-β) levels. Furthermore, they increased the frequency of larger cells with nuclear alterations, the levels of oxidative markers, and interleukin 1β, a marker of the Senescence-Associated Secretory Phenotype (SASP). However, the mitochondrial O2⁻ imbalance caused by ROT led to more pronounced alterations compared to PQT. These findings support the hypothesis that the existence of CAT components, such as the O2⁻ anion, plays a significant role in cellular aging.

PEAR1 Promotes Myoblast Proliferation Through Notch Signaling Pathway.

PEAR1, also known as platelet endothelial aggregation receptor 1, is known to play a crucial role in the migration and differentiation of muscle satellite cells (MuSCs). However, its specific effects on skeletal muscle development and regeneration require further exploration. In this study, the expression of PEAR1; the proliferation marker proteins of Pax7, CCNB1, and PCNA; and the key molecules of N1-ICD, N2-ICD, and Hes1 were all increased gradually during the process of C2C12 cell proliferation. Furthermore, Western blotting and EdU results showed that when PEAR1 was over-expressed or inhibited, the proliferation status of C2C12 cell was increased or reduced respectively. This implied that PEAR1 could regulate myoblast proliferation and might be relate to Notch cell signaling pathway. A subsequent immunoprecipitation experiment result showed that the interaction between PEAR1 and Notch1 or Notch2, respectively. Then Western blotting and EdU results showed that the proliferation of C2C12 cell was inhibited under the treatment of Notch signaling pathway inhibitor RIN1. Meanwhile, the proliferation capacity of C2C12 cell could not be improved by treatment with RIN1 even though PEAR1 was over-expressed. These results showed that PEAR1 may regulated C2C12 cell proliferation though Notch signaling pathway. Additionally, a mouse model of muscle injury repair injected with bupivacaine hydrochloride was established in this study. Immunohistochemistry results exhibited that PEAR1 may regulate skeletal muscle post-injury regeneration relevant to Notch1 and Notch2 in different patterns. These findings provide valuable insights into the potential involvement of PEAR1 in skeletal muscle development and post-injury regeneration.

Theabrownin/whey protein isolate complex coacervate strengthens C2C12 cell proliferation via modulation of energy metabolism and mitochondrial apoptosis.

Theabrownin (TB)-whey protein isolate (WPI) complex coacervates (TW) were firstly prepared to investigate the regulatory effects on skeletal muscle. The binding of TB to WPI reached saturation with the strongest electrostatic interaction at the ratio of 10:1. The formation of TW was driven by electrostatic interactions with the aid of hydrogen bonding and hydrophobic interactions, and the digestion behavior of TW was investigated based on in vitro gastrointestinal and CaCO2 cell models. The regulatory effect of TW on muscle cells was investigated by C2C12 cell assay. Cell cycle analysis showed that TW promoted the transition of skeletal muscle cells from proliferative state to differentiated state. Immunofluorescence and gene expression revealed that TW positively regulated myogenic regulatory factors, contributing to myofiber formation. Moreover, TW activated the intracellular TCA cycling and oxidative phosphorylation, providing energy for skeletal muscle regeneration and repair. Mechanistically, TW inhibited the release of cytochrome C from mitochondria to cytoplasm through the Bcl-2/Cytochrome C/Cleaved-Caspase-3 pathway, exhibiting a protective effect on skeletal muscle cells. In the future, the molecular mechanism of TW enhancing skeletal muscle function should be validated through aging animal models and clinical trials and expand its therapeutic application for muscle health in functional food and dietary supplements.

Preoperative prediction of Ki-67 expression in hepatocellular carcinoma by spectral imaging on dual-energy computed tomography (DECT).

The Ki-67 expression level, which represents the proliferation status of cells, serves as a prognostic marker for hepatocellular carcinoma (HCC); however, the immunochemistry method currently used to evaluate Ki-67 is invasive and is not suitable for patients who have lost the chance for surgery, such as those with advanced tumors or those with other serious organic diseases. This study aimed to investigate the value of quantitative dual-energy computed tomography (DECT) parameters in predicting the expression level of Ki-67 in HCC. This study analyzed the data of 123 consecutive patients with HCC who underwent both Ki-67 immunohistochemistry analysis and two-phase contrast-enhanced DECT imaging. The patients were divided into the following two groups based on the positive rate of Ki-67 (Ki-67%): the high-expression group (Ki-67% >20%, n=74); and the low-expression group (Ki-67% ≤20%, n=49). The computed tomography (CT) values in the 130 and 140 keV monochromatic energy images (HU130-140keV), normalized effective atomic number (NeffZ), fat density (Dfat), and water density (Dwater) were measured and calculated at the arterial phase (AP) and portal venous phase (PVP). A Spearman correlation coefficient analysis, a comparison of parameters between groups, a receiver operating characteristic (ROC) curve analysis for evaluating predictive efficacy, and a multivariable logistic regression analysis were conducted. The NeffZ-AP, HU130 keV-PVP, HU140 keV-PVP, Dfat-PVP, and Dwater-PVP were positively correlated with the Ki-67% (all P<0.05), and the DECT parameter values in the high-expression group were significantly higher than those in the low-expression group (all P<0.05). The Dwater-PVP [odds ratio (OR) =1.353, 95% confidence interval (CI): 1.204-1.521, P<0.001] and tumor diameter (OR =1.258, 95% CI: 1.08-1.465, P=0.003) were independent predictive factors for high Ki-67 expression. Dwater-PVP had the highest predictive efficacy with an area under the curve (AUC) of 0.810. The multivariable analysis combining the DECT parameters and morphological characteristics improved the predictive efficacy of the model in predicting high Ki-67 expression (AUC =0.857). DECT provides a non-invasive method to evaluate the proliferation status of HCC cells, and the predictive efficacy of high Ki-67 expression could be improved by combining DECT parameters and morphologic features.

Dysregulated RNA splicing induces regeneration failure in alcohol-associated liver disease.

Individuals with progressive liver failure are at a high risk of mortality without liver transplantation. However, our understanding of derailed regenerative responses in failing livers is limited. Here, we performed comprehensive multi-omic profiling of healthy and diseased human livers using bulk and single-nucleus RNA-plus ATAC-seq. We report that hepatic immune milieu alterations in alcohol-associated liver disease (ALD) prevent hepatocytes from transitioning to a proliferative progenitor-like state, trapping them into an unproductive intermediate state. We discovered striking changes in RNA binding protein (RBP) expression, particularly ESRP, PTBP, and SR families, that cause misregulation of developmentally controlled RNA splicing in ALD. Our data pinpoint ESRP2 as a pivotal disease-sensitive RBP and support a causal role of its deficiency in ALD pathogenesis. Notably, splicing defects in ESRP2-targets Tcf4 and Slk , amongst others, directly alter their nuclear localization and activities, disrupting WNT and Hippo signaling pathways, which are critical for normal liver regeneration. We demonstrate that changes in stromal cell populations enrich failing ALD livers with TGF-β, which suppresses ESRP2-driven epithelial splicing program and replaces functional parenchyma with quasi-progenitor-like cells lacking liver-specific functions. This unprecedented account of transcriptional and post-transcriptional dysregulation in ALD suggests that targeting misspliced RNAs could improve recovery and serve as biomarkers for poor ALD outcomes.

Assessment of in vivo and in vitro anti-tumoral effects of Lycium barbarum extract on Ehrlich ascites tumor cells: histopathology, DNA damage and AgNOR.

Natural product research has an exciting and glorious past that spans over millennia. Accordingly, natural products mediated inhibition of carcinogenesis by mechanistic modulation of deregulated signaling pathways has revolutionized the field of translational oncology. Lycium barbarum has antioxidant and anticarcinogenic effects. The antioxidant activity of the extract and its effect on Ehrlich ascites tumor (EAT) were investigated using in vivo and in vitro techniques. EAT cells were injected into Balb/C mice to create stock mice. EAT cells withdrawn from stock mice were used in equal volumes in the studies. The in vivo study consisted of control and treatment groups (200 mg/kg fractions above and below 50 kDa of extracts). The liver tissues were evaluated for histopathological (H&E), DNA damage (Comet assay), and proliferation (AgNOR staining) status. The in vitro study consisted of control and treatment groups (1500 and 2000 µg/ml of extracts). Cell viability and apoptosis were evaluated. As a result, a decrease in the adhesion of EAT cells, and decreased DNA damage were observed in mice intraperitoneally administered with the fractions of Lycium barbarum. The extracts both below and above 50 kDa increased apoptotic death in cancer cells. The extract above 50 kDa was more active than those below 50 kDa. Lycium barbarum consumption may be effectual in preventing cancer formation and slowing the progression of cancer.

Unveiling dynamic hepatocyte plasticity in HepaRG cells with a dual CYP reporter system.

Primary hepatocytes are widely utilized for investigating drug efficacy and toxicity, yet variations between batches and limited proliferation capacity present significant challenges. HepaRG cells are versatile cells, capable of maintaining an undifferentiated state and differentiating through dimethyl sulfoxide treatment, allowing for molecular analysis of hepatocyte plasticity. To elucidate the underlying molecular mechanisms of HepaRG cell plasticity, we used CYP3A4G/7R HepaRG cells engineered to express DsRed under the control of the fetus-specific CYP3A7 gene and EGFP under the adult-specific CYP3A4 gene promoter. In time-lapse imaging of CYP3A4G/7R HepaRG cells, we observed CYP3A7-DsRed expression transitioning from negative to positive during proliferation period and CYP3A4-GFP expression activating during differentiation. The de-differentiation potency of differentiated CYP3A4G/7R HepaRG cells was assessed using inhibitors and cytokines. It was found that Y-27632 (Y), A-83-01 (A), and CHIR99021 (C) (collectively referred to as YAC), which are known to promote liver regeneration in mice, did not induce CYP3A7-DsRed expression. Instead, these inhibitors increased CYP3A4-GFP expressing population. Furthermore, CHIR99021 alone increased CYP3A4-GFP-positive cells, while Wnt3a treatment increased CYP3A7-DsRed-positive cells, suggesting that Wnt signaling plays distinct roles in HepaRG cells. It was apparent that de-differentiated cells had increased CYP3A4 activity after a second round of differentiation, compared to differentiated cells after the first round. Transcriptomic analysis of HepaRG cells revealed distinct profiles between proliferative, differentiated, and de-differentiated states, highlighting their robust plasticity. Notably, hepatoblastic cells de-differentiated by YAC or C displayed transcriptome patterns similar to undifferentiated cells, whereas CYP3A7-DsRed and CYP3A4-GFP exhibited expression patterns different from those of undifferentiated cells. These findings underscore the dynamic nature of HepaRG cells while cautioning against solely relying on CYP3 family gene expression as a marker of differentiation.

From the genome's perspective: Bearing somatic retrotransposition to leverage the regulatory potential of L1 RNAs.

Transposable elements (TEs) are mobile genomic elements constituting a big fraction of eukaryotic genomes. They ignite an evolutionary arms race with host genomes, which in turn evolve strategies to restrict their activity. Despite being tightly repressed, TEs display precisely regulated expression patterns during specific stages of mammalian development, suggesting potential benefits for the host. Among TEs, the long interspersed nuclear element (LINE-1 or L1) has been found to be active in neurons. This activity prompted extensive research into its possible role in cognition. So far, no specific cause-effect relationship between L1 retrotransposition and brain functions has been conclusively identified. Nevertheless, accumulating evidence suggests that interactions between L1 RNAs and RNA/DNA binding proteins encode specific messages that cells utilize to activate or repress entire transcriptional programs. We summarize recent findings highlighting the activity of L1 RNAs at the non-coding level during early embryonic and brain development. We propose a hypothesis suggesting a mutualistic relationship between L1 mRNAs and the host cell. In this scenario, cells tolerate a certain rate of retrotransposition to leverage the regulatory effects of L1s as non-coding RNAs on potentiating their mitotic potential. In turn, L1s benefit from the cell's proliferative state to increase their chance to mobilize.

RNA/DNA Binding Protein TDP43 Regulates DNA Mismatch Repair Genes with Implications for Genome Stability.

TAR DNA-binding protein 43 (TDP43) is increasingly recognized for its involvement in neurodegenerative diseases, particularly amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). TDP43 proteinopathy, characterized by dysregulated nuclear export and cytoplasmic aggregation, is present in most ALS/FTD cases and is associated with a loss of nuclear function and genomic instability in neurons. Building on prior evidence linking TDP43 pathology to DNA double-strand breaks (DSBs), this study identifies a novel regulatory role for TDP43 in the DNA mismatch repair (MMR) pathway. We demonstrate that depletion or overexpression of TDP43 affects the expression of key MMR genes, including MLH1, MSH6, MSH2, MSH3, and PMS2. Specifically, TDP43 modulates the expression of MLH1 and MSH6 proteins through alternative splicing and transcript stability. These findings are validated in ALS mice models, patient-derived neural progenitor cells and autopsied brain tissues from ALS patients. Furthermore, MMR depletion showed a partial rescue of TDP43-induced DNA damage in neuronal cells. Bioinformatics analysis of TCGA cancer database reveals significant correlations between TDP43 and MMR gene expressions and mutational burden across various cancer subtypes. These results collectively establish TDP43 as a critical regulator of the MMR pathway, with broad implications for understanding the genomic instability underlying neurodegenerative and neoplastic diseases.

Evaluation of the immunomodulatory effects of a novel synbiotic made of combined use of probiotic-prebiotic-Chinese traditional herbs

BackgroundRecent studies have discussed the preferred activity of synbiotics, which are a combination of probiotics and prebiotics. This study formulates a novel synbiotic that includes Chinese traditional herbs and evaluates its immunomodulatory function. PurposeThis study aims to systematically investigate the effects of synbiotics on immune function using a cyclophosphamide-induced immunosuppressed mouse model. Study designA literature-based approach was used to selectively identify medicinal herbs with immunomodulatory properties. Commercially available probiotics were used to investigate the immunomodulatory functions and potential mechanisms associated with the combination of Chinese medicinal herbs and prebiotics as carbon substrates. MethodsThe immunomodulatory effect of the synbiotic will be assessed by measuring indicators such as body weight, organ index, immunoglobulins, cytokines, splenic lymphocyte proliferation status, NK cell cytotoxicity and gut microbiota. ResultsOur results demonstrated that the synbiotic significantly restored the decreased thymus index and the increased spleen index induced by immune deficiency in mice. Among them, the formula of the synbiotics significantly improves the thymus organ index of immune organs from 0.068 to 0.126, which is also significantly superior compared to other single components. Additionally, it significantly enhanced the proliferative capacity of splenic lymphocytes and the cytotoxic activity of natural killer (NK) cells in immune-deficient mice. Furthermore, the synbiotics enhanced the richness and diversity of the gut microbiota, rectifying the dysbiosis in gut microbial composition caused by immune deficiency. It decreased the relative abundance of Firmicutes from 67.41 % to 44.06 %, while increasing the relative abundance of Bacteroidota from 30.42 % to 53.77 %. It restored the disrupted gut microbial structure induced by cyclophosphamide damage, thereby ameliorating the disrupted intestinal microbial community associated with immune deficiency. ConclusionThe study demonstrated that the synbiotic was capable of modulating immune function, as evidenced by an increase in organ index, cellular immunity, and the ability to modulate intestinal flora disorders. Furthermore, the synbiotic demonstrated superior performance compared to other single components.

AI-powered precision: breast carcinoma diagnosis through digital proliferation index (Ki-67) assessment in pathological anatomy

PurposeThe primary objective of the study is to enhance the accuracy and efficiency of assessing the proliferation index in cancer cells, specifically focusing on the role of Ki-67. The purpose is to address the limitations of traditional visual assessments conducted by pathologists by integrating AI technologies, particularly deep learning. By accurately computing the percentage of Ki-67-labeled cells, the research aims to streamline the diagnostic process, reduce subjectivity and contribute to the advancement of diagnostic precision in pathological anatomy.Design/methodology/approachThe research employs a methodological approach that integrates Ki-67, a non-histone nuclear protein, as a vital biomarker for assessing the proliferative status of cancer cells. Given the challenges associated with traditional visual assessments by pathologists, including inter- and intra-observer variability and time-consuming efforts, the study adopts a novel methodology leveraging artificial intelligence (AI) solutions. Deep learning is applied to precisely calculate the percentage of Ki-67-labeled cells. The process involves pathologists delineating the tumor area at x40 magnification, enabling the segmentation of various cell types (positive, negative and tumor-infiltrating lymphocytes). The subsequent percentage calculation enhances efficiency and minimizes subjectivity in the diagnostic process.FindingsDespite inherent errors, the research findings indicate that the model surpasses existing benchmarks, showcasing superior accuracy in terms of average error measurement. The comparison with diverse datasets and benchmarking against pathologists’ diagnoses contributes empirical evidence to support the effectiveness of the AI-based model in accurately computing the percentage of Ki-67-labeled cells. These findings signify a noteworthy advancement in diagnostic methodologies and reinforce the potential of AI technologies in improving the precision of cancer diagnostics within the realm of pathological anatomy.Originality/valueThe research contributes to the field by introducing an innovative approach that combines Ki-67 as a biomarker and AI technologies for improved diagnostic precision. The originality lies in the utilization of deep learning to calculate the percentage of labeled cells, mitigating the challenges associated with manual assessments. The validation of the model against diverse datasets and benchmarking against pathologists’ diagnoses demonstrates its superior accuracy, highlighting the value of integrating AI in pathological anatomy for enhanced diagnostic outcomes. The study represents a significant stride in original research, offering novel insights and methodologies in the pursuit of more precise cancer diagnostics.

Oral N-acetylcysteine ameliorates liver fibrosis and enhances regenerative responses in Mdr2 knockout mice

Cholangiopathies are poorly understood disorders with no effective therapy. The extrahepatic biliary tree phenotype is less studied compared to the intrahepatic biliary injury in both human disease and Mdr2−/− mice, the established cholestatic mouse model. This study aimed to characterize the extra hepatic biliary tree of Mdr2−/− mice at various ages and to determine if injury can be repaired with the antioxidant and glutathione precursor N-acetyl-L-Cysteine treatment (NAC). We characterized extra hepatic bile ducts (EHBD)s at various ages from 2 to 40 weeks old FVB/N and Mdr2−/− mice. We examined the therapeutic potential of local NAC ex vivo using EHBD explants at early and late stages of injury; and systematic therapy by in vivo oral administration for 3 weeks. EHBD and liver sections were assessed by histology and immunofluorescent stains. Serum liver enzyme activities were analyzed, and liver spatial protein expression analysis was performed. Mdr2−/− mice developed progressive EHBD injury, similar to extrahepatic PSC. NAC treatment of ex vivo EHBD explants led to improved duct morphology. In vivo, oral administration of NAC improved liver fibrosis, and decreased liver enzyme activities. Spatial protein analysis revealed cell-type specific differential response to NAC, collectively indicating a transition from pro-apoptotic into proliferative state. NAC treatment should be further investigated as a potential therapeutic option for human cholangiopathies.

PH-sensing GPR68 inhibits vascular smooth muscle cell proliferation through Rap1A.

Phenotypic transformation of vascular smooth muscle (VSM) from a contractile state to a synthetic, proliferative state is a hallmark of cardiovascular disease (CVD). In CVD, diseased tissue often becomes acidic from altered cellular metabolism secondary to compromised blood flow, yet the contribution of local acid/base imbalance to the disease process has been historically overlooked. In this study, we examined the regulatory impact of the pH-sensing G protein-coupled receptor GPR68 on vascular smooth muscle (VSM) proliferation in vivo and in vitro in wild-type (WT) and GPR68 knockout (KO) male and female mice. Arterial injury reduced GPR68 expression in WT vessels and exaggerated medial wall remodeling in GPR68 KO vessels. In vitro, KO VSM cells showed increased cell-cycle progression and proliferation compared with WT VSM cells, and GPR68-inducing acidic exposure reduced proliferation in WT cells. mRNA and protein expression analyses revealed increased Rap1A in KO cells compared with WT cells, and RNA silencing of Rap1A reduced KO VSM cell proliferation. In sum, these findings support a growth-inhibitory capacity of pH-sensing GPR68 and suggest a mechanistic role for the small GTPase Rap1A in GPR68-mediated VSM growth control. These results shed light on GPR68 and its effector Rap1A as potential targets to combat pathological phenotypic switching and proliferation in VSM.NEW & NOTEWORTHY Extracellular acidosis remains an understudied feature of many pathologies. We examined a potential regulatory role for pH-sensing GPR68 in vascular smooth muscle (VSM) growth in the context of CVD. With in vivo and in vitro growth models with GPR68-deficient mice and GPR68 induction strategies, novel findings revealed capacity of GPR68 to attenuate growth through the small GTPase Rap1A. These observations highlight GPR68 and its effector Rap1A as possible therapeutic targets to combat pathological VSM growth.

High-parametric protein maps reveal the spatial organization in early-developing human lung

The respiratory system, including the lungs, is essential for terrestrial life. While recent research has advanced our understanding of lung development, much still relies on animal models and transcriptome analyses. In this study conducted within the Human Developmental Cell Atlas (HDCA) initiative, we describe the protein-level spatiotemporal organization of the lung during the first trimester of human gestation. Using high-parametric tissue imaging with a 30-plex antibody panel, we analyzed human lung samples from 6 to 13 post-conception weeks, generating data from over 2 million cells across five developmental timepoints. We present a resource detailing spatially resolved cell type composition of the developing human lung, including proliferative states, immune cell patterns, spatial arrangement traits, and their temporal evolution. This represents an extensive single-cell resolved protein-level examination of the developing human lung and provides a valuable resource for further research into the developmental roots of human respiratory health and disease.

The value of multiple diffusion metrics based on whole-lesion histogram analysis in evaluating the subtypes and proliferation status of non-small cell lung cancer.

Limited studies have explored the utility of whole-lesion histogram analysis in discerning the subtypes and proliferation status of non-small cell lung cancer (NSCLC), despite its potential to provide comprehensive tissue assessment through the computation of additional quantitative metrics. This study sought to assess the significance of intravoxel incoherent motion (IVIM) and diffusion kurtosis imaging (DKI) histogram parameters in discriminating between squamous cell carcinoma (SCC) and adenocarcinoma (AC), and to examine the correlation of each parameter with the proliferative marker Ki-67. Patients with space-occupying lesions detected by chest CT examination and with further routine MRI, DKI and IVIM functional sequence scans were enrolled. Based on the pathological results, seventy patients with NSCLC were selected and divided into AC and SCC groups. Histogram parameters of IVIM (D, D*, f) and DKI (Dapp, Kapp) were calculated, and the Mann-Whitney U test or independent samples t test was used to analyze the differences in each histogram parameter of the SCC and AC groups. Receiver operating characteristic (ROC) curves were used to evaluate the diagnostic performance of the histogram parameters. The correlation coefficient between histogram parameters and Ki-67 was calculated using Spearman's or Pearson's methods. The D 10th percentile, D 90th percentile, D mean, D median, Dapp 10th percentile, Dapp 90th percentile, Dapp mean, Dapp median, Dapp skewness, Dapp SD of the AC groups were significantly higher than those of the SCC groups, while the Kapp entropy and Kapp SD of the SCC groups were significantly higher than those of the AC groups. All the above differences were statistically significant (all P < 0.05). ROC curve analysis revealed that Dapp mean showed the best performance for differentiating AC from SCC lesions, with an area under the ROC curve of 0.832 (95% confidence interval [CI]: 0.707-0.919). But there was no statistically significant difference in diagnostic efficacy compared to other histogram parameters (all P>0.05). Dapp 90thpercentile, Dapp mean, Kapp skewnes showed a slight negative correlation with Ki-67 expression (r value -0.340, -0.287, -0.344, respectively; P< 0.05), while the other histogram parameters showed no significant correlation with Ki-67 (all P > 0.05). Our study demonstrates the utility of IVIM and DKI histogram analyses in differentiating NSCLC subtypes, particularly AC and SCC. Correlations with the Ki-67 index suggest that Dapp mean, Dapp 90th percentile, and Kapp skewness may serve as markers of tumor aggressiveness, supporting their use in NSCLC diagnosis and treatment planning.

Non-genetic differences underlie variability in proliferation among esophageal epithelial clones.

Individual cells grown in culture exhibit remarkable differences in their growth, with some cells capable of forming large clusters, while others are limited or fail to grow at all. While these differences have been observed across cell lines and human samples, the growth dynamics and associated cell states remain poorly understood. In this study, we performed clonal tracing through imaging and cellular barcoding of an in vitro model of esophageal epithelial cells (EPC2-hTERT). We found that about 10% of clones grow exponentially, while the remaining have cells that become non-proliferative leading to a halt in the growth rate. Using mathematical models, we demonstrate two distinct growth behaviors: exponential and logistic. Further, we discovered that the propensity to grow exponentially is largely heritable through four doublings and that the less proliferative clones can become highly proliferative through increasing plating density. Combining barcoding with single-cell RNA-sequencing (scRNA-seq), we identified the cellular states associated with the highly proliferative clones, which include genes in the WNT and PI3K pathways. Finally, we identified an enrichment of cells resembling the highly proliferative cell state in the proliferating healthy human esophageal epithelium.

Accelerated Wound Healing of Tetrahedral-Framework Nucleic Acid Nanozymes with High Penetration and Antioxidant Capacity.

The wound repair process usually leads to a non-functioning mass of fibrotic tissue because of the oxidative imbalance of deep tissue layers. However, how to improve the penetration of active ingredients into deeper layers and regulate oxidative imbalances to create a regenerative microenvironment still remains a challenge. In this study, we designed a novel tetrahedral-framework nucleic acid (tFNA) nanozyme that could penetrate the skin/mucosa barrier as deep as 450 μm within 24 h. We also demonstrated the protective role of tFNAs on the mitochondrial structural and functional integrity and inhibition of reactive oxygen species production to repair oxidative imbalances through ERK1/2-Nrf2-HO-1 during repair processes. It was found that the proliferative state and the migration ability of postburn cells in vitro were accelerated, and the early closure of wounds in vivo was significantly promoted. This study therefore provides a promising strategy to efficiently regulate the oxidative imbalances in the deep layers of the skin during wound healing.

Sinonasal adenoid cystic carcinoma: preoperative apparent diffusion coefficient histogram analysis in prediction of prognosis and Ki-67 proliferation status.

To investigate the value of preoperative apparent diffusion coefficient (ADC) histogram analysis in predicting the prognosis of patients with sinonasal adenoid cystic carcinoma (ACC) and the correlation between ADC histogram parameters and Ki-67 labeling index (LI). The study enrolled 66 patients with sinonasal ACC who were surgically resected and confirmed by histopathology. The disease-free survival (DFS) was evaluated with clinical-pathologic and radiologic characteristics using the Cox proportion hazard model. Spearman correlation analysis was used to evaluate the correlation between ADC histogram parameters and Ki-67 LI. The predictive performance of ADC histogram parameters for Ki-67 LI was assessed using the receiver operating characteristic (ROC) curve. Multivariable analysis showed Ki-67 LI (hazard ratio: 9.279; 95% confidence interval 1.099-78.338; P = 0.041) and ADCskewness (hazard ratio: 5.942; 95% confidence interval 1.832-19.268; P = 0.003) were significant independent predictors of DFS. The combination of these two variables achieved the predictive ability with a C-index of 0.717 (95% confidence interval 0.607-0.826). ADCmean and all ADC percentiles (10th, 50th, and 90th) significantly and inversely correlated with Ki-67 LI of ACC (Correlation coefficients = -0.574 to -0.591, Ps < 0.001). Among the ADC histogram parameters, the ADC50th showed superior performance for the differentiation of the high from low Ki-67 LI groups with an area under the curve (AUC) of 0.834 and an accuracy of 80.30%. ADC histogram analysis had predictive value for DFS and Ki-67 LI, which may be a valuable biomarker for prognosis and proliferation status for ACC in clinical practice.

Exploring the Role of Sigma Receptors in the Treatment of Cancer: A Narrative Review.

This study investigated the association of sigma receptors (SRs) and their selective ligands (because the molecular characteristics of the same SRs, particularly sigma-2 receptor {S2R}, are not completely clear) in carcinogenesis, their potential use as antitumor agents, and their great utility in tumor imaging.The ion channels and transporters enhance the cell's ability to adapt to the metabolic conditions encountered in the tumor tissue. The high expression of SRs in the proliferating cells compared with those at rest indicates that this is a significant clinical biomarker for determining the proliferative status of solid tumors using functional PET imaging techniques. The association of SRs in the pathophysiology of cancer cells is a result of the high concentration of S1R and S2R binding sites observed in various tumor cell lines and tissues. It would also be remarkable to determine if SRs are involved in metastasis and other metastatic cell behaviorssuch as adhesion, secretion, motility, and penetration. An absolute challenge for research in this field is to develop an integrated model that describes the molecular mechanisms of sigma receptors, incorporating their known biological and pathophysiological roles.