High Proliferation Research Articles

Mechanosensitive Ion Channels: Their Physiological Importance and Potential Key Role in Cancer.

Mechanosensitive ion channels comprise a broad group of proteins that sense mechanical extracellular and intracellular changes, translating them into cation influx to adapt and respond to these physical cues. All cells in the organism are mechanosensitive, and these physical cues have proven to have an important role in regulating proliferation, cell fate and differentiation, migration and cellular stress, among other processes. Indeed, the mechanical properties of the extracellular matrix in cancer change drastically due to high cell proliferation and modification of extracellular protein secretion, suggesting an important contribution to tumor cell regulation. In this review, we describe the physiological significance of mechanosensitive ion channels, emphasizing their role in cancer and immunity, and providing compelling proof of the importance of continuing to explore their potential as new therapeutic targets in cancer research.

Metformin suppresses cardiac fibroblast proliferation under high-glucose conditions via regulating the mitochondrial complex I protein Grim-19 involved in the Sirt1/Stat3 signaling pathway.

Hyperglycemia associated with myocardial oxidative stress and fibrosis is the main cause of diabetic cardiomyopathy. Currently, no approved drug is available for preventing or treating diabetes-induced cardiac fibrosis. Metformin has been reported to improve glycemic control and ameliorate diabetic cardiomyopathy. This study aimed to investigate the effects and mechanism of metformin on diabetes-induced cardiac fibrosis and high glucose-induced proliferation of cardiac fibroblasts (CFs). In this study, db/db mice were treated with metformin [250mg/kg⋅d, gavage]. CFs were cultured in high-glucose medium to mimic an in vitro diabetes model and then subjected to treatment with or without metformin. Cardiac fibrosis was analyzed using immunohistochemistry, Masson's trichrome staining, and Western blot analysis. Cell Counting Kit-8 (CCK-8) assays and cell colony formation assays were used to examine cell proliferation capacity. Transwell and scratch-wound assays were used to detect the migration ability of CFs. Retinoid-interferon-induced mortality-19 (Grim-19), sirtuin1 (Sirt1), and signal transducer and activator of transcription 3 (Stat3) were detected using Western blot analysis. The genes downstream of the Stat3 pathway were detected using quantitative reverse transcription PCR (qRT‒PCR). Metformin treatment markedly attenuated cardiac fibrosis in db/db mice and the proliferation and migration of CFs under high-glucose conditions. Mechanistically, we found an intersection between metformin and Grim-19 using bioinformatics. Metformin was found to suppress the expression of p-Stat3 and elevate the expression of mitochondrial complex I protein Grim-19 and Sirt1, thus inhibiting the proliferation and migration of CFs under high-glucose conditions. Our data suggested that metformin inhibited the proliferation and migration of CFs by regulating the expression of mitochondrial complex I Grim-19 protein involved in the Sirt1/Stat3 signaling pathway under high-glucose conditions, thus providing new ideas for treating diabetes-induced cardiac fibrosis.

Kedilerin Yassı Hücreli Karsinomlarında İntermedier Filamentler, p53 Geni, Hücre Proliferasyonu, Metastaz ve Apoptozis

In this study, intermediate filaments, Pan Cytokeratin (Pan CK), Vimentin, Desmin and S-100 were evaluated to reveal the cellular origin of feline SCCs. Alpha-Smooth Muscle Actin (α-SMA) for cancer-related fibroblasts (CAFs) in the tumor microenvironment, and p53, a tumor suppressor gene, were investigated. Proliferating Cell Nuclear Antigen (PCNA) expression was evaluated for the cell proliferation index. Matrix Metalloproteinase-9 (MMP-9) immunoreactivity was evaluated for the metastasis and invasion capacity. In addition, it is aimed to reveal the expressions of proapoptotic Bax gene, antiapoptotic Bcl-2 gene, caspase-dependent pathway Caspase-3 and caspase-independent pathway Apoptosis Inducing Factor (AIF) for apoptosis mechanism. Biopsy samples taken from 7 cats brought to Department of Pathology for routine histopathological examination were used in this study. Tumor tissue samples were fixed in 10% formaldehyde solution. Serial sections of 5 μm thickness were taken from the paraffin blocks prepared after routine tissue follow-up procedures. Hematoxylin & Eosin (H&E) staining was performed on the sections. Avidin-Biotin Peroxidase-Technique (ABC) was used as immunohistochemical staining. It was determined that the tumors had epithelial-mesenchymal transition, exhibited a very high proliferation index, had p53 mutation, and showed low metastasis/high invasion capacity. It was revealed that Bax/Bcl-2 ratio increased in favor of proapoptotic Bax, and caspase-independent apoptosis was more dominant than caspase-dependent apoptosis.

Mesenchymal Stem Cells Derived from Human Periapical Cysts and Their Implications in Regenerative Medicine.

Mesenchymal stem cells currently play an important role in the tissue engineering field in developing new regenerative approaches. The oral cavity is a rich source of mesenchymal stem cells, and introducing the use of dental stem cells, characterized by a multilineage differentiation potential, immunomodulatory activity and repair capacity, offers a good perspective for clinical dentistry. Human periapical cyst mesenchymal stem cells (hPCy-MSCs) represent a new category of dental stem cells, being collected from pathological tissue and exhibiting MSCs-like properties. As studies have described, these new identified cells possess the same characteristics as those described in MSCs, exhibiting plasticity, a high proliferation rate and the potential to differentiate into osteogenic, adipogenic and neural lineages. Reusing the biological tissue that is considered pathologic offers a new perspective for the development of further clinical applications. The identification and characterization of MSCs in the human periapical cysts allows for a better understanding of the molecular interactions, the potential healing capacity and the mechanisms of inducing the local osteogenic process, integrated in the microenvironment. Although their involvement in regenerative medicine research is recent, they exhibit important properties that refer them for the development of clinical applications in dentistry.

Development of a low-cost quad-extrusion 3D bioprinting system for multi-material tissue constructs

This study presents the development and characterization of a low-cost bioprinting system with a compact low-profile quad-extrusion bioprinting head for producing multi-material tissue constructs. The system, created by modifying an off-the-shelf three-dimensional (3D) printer, enables larger print volumes compared to extant systems. Incorporating gelatin methacrylate (GelMA) as a bioink model, the bioprinting system was systematically tested with two different printing techniques, namely the traditional in-air printing (IAP) mode along with an emerging support bath printing (SBP) paradigm. Structural fidelity was assessed by comparing printed structures under different conditions to the computer-aided design (CAD) model. To evaluate biological functionality, a placental model was created using HTR-8 trophoblasts known for their invasive phenotype. Biological assays of cell viability and invasion revealed that the cells achieved high cell proliferation rates and had over 93% cell viability for a 3-day incubation period. The multi-compartmental 3D-bioprinted in vitro placenta model demonstrates the potential for studying native cell phenotypes and specialized functional outcomes enabled by the multi-material capability of the quad-extrusion bioprinter (QEB). This work represents a significant advancement in bioprinting technology, allowing for the printing of complex and highly organized tissue structures at scale. Moreover, the system’s total build cost is only US$ 297, making it an affordable resource for researchers.

Identification of a novel carboxylesterase dominantly expressed in the booklouse, Liposcelis bostrychophila

Booklice are tiny insect pests commonly found indoors and characterized by high proliferation ability. Booklice contamination in stored foods causes profound food loss and possibly allergic reactions after ingestion; thus, booklice are known as stored-food pests. Therefore, the safe extermination of booklice would solve food loss and allergic concerns. Organophosphates (OPs) are used as harmless insecticides against various pests; nonetheless, cases of OP poisoning are reported worldwide. Therefore, considering protecting stored-food products, OP doses should be minimal. It is generally known that carboxylesterase-mediated inactivation of OPs decreases the efficacy of OPs, suggesting that an inhibitor of carboxylesterase could be a synergist in reducing the effective dose of OP. Herein, I report the molecular cloning of a novel carboxylesterase dominantly expressed in a representative species of indoor booklice, Liposcelis bostrychophila, designated Liposcelis bostrychophila carboxylesterase 1 (LBCE1). The recombinant protein of LBCE1 was shown to have an elastase activity. Furthermore, using the 3D-structural model of LBCE1, its possible binding to OP insecticides was predicted in silico. These results suggest that a specific LBCE1 inhibitor may serve as a synergist to better control booklice.

Comparison of Ki67 Proliferation Index in Gastrointestinal Non-Hodgkin Large B-Cell Lymphomas: The Conventional Method of Evaluation or AI Evaluation?

The most common NHL subtype in SEEU is DLBCL (39%), and it manifests with a variety of cellular morphologies and a high proliferation index. Also, the GI tract is the most common site of extranodal NHLs, and most NHLs involving the GI tract are of B-cell lineage, of which diffuse large B-cell lymphoma is the most common subtype, irrespective of location. The last few years have seen digital pathology as a vital technology that has a positive impact on diagnostics, but studies on the use of DP for lymphoma identification, however, are still restricted to only determining whether a tumor is present or absent. Using the example of cases of malignant NHL, we aim to investigate the diagnostic utility of DP using QuPath software in evaluating the proliferation index and the prognostic significance and to show that improved visualization and analysis contribute to the convergence of these complementary diagnostic modalities for lymphomas. The average proliferation index (Ki67) was 58.33% with values between 10% and 85%. After the stratification of the cases, an increased proliferation index was observed in the majority of cases (53.33%), and this aspect was associated with the advanced age of the patients (p = 0.045). Visual assessment provides lower Ki67 values than automated digital image analysis. However, the agreement coefficient between the conventional method and the AI method indicates an excellent level of reliability (ICC1-0.970, ICC2-0.990). The multivariate analysis revealed that in the cases where the proliferation index Ki67 is high (˃70%), the IPI score represents an important risk factor predicting mortality (HR = 10.597, p = 0.033).

Differences in clinical outcomes between HER2-negative and HER2-positive luminal B breast cancer.

The clinical features and prognosis of breast cancer can vary widely, depending on the molecular subtype. Luminal B breast cancers are usually either estrogen receptor-positive and/or progesterone receptor-positive with high proliferation of Ki67 index, or HER2 positive (HER2+). The authors compared the clinicopathologic factors and survival rates of different subtypes of luminal B breast cancer according to HER2 status. Between 2009 and 2013, 1131 cases of breast cancer were reviewed and characterized as 1 of 4 different molecular subtypes based on their immunohistochemical results: luminal A, luminal B, HER2+, or triple-negative breast cancer. From these, luminal B breast cancers were extracted and the clinical features and prognosis of the HER2- and the HER2 + subtypes were compared. Survival differed significantly based on the molecular subtype regardless of whether or not the patient received treatment with neoadjuvant chemotherapy. While patients with HER2- luminal B breast cancer who received neoadjuvant chemotherapy had better prognoses, patients with HER2 + luminal B breast cancer who did not receive neoadjuvant chemotherapy had better prognoses. Luminal B breast cancers showed different clinical outcomes and survival rates according to HER2 gene overexpression type. Physicians should consider these results when they establish a treatment strategy.

Proteomics-based clustering of lung adenocarcinoma identifies three subtypes with significantly different clinical and molecular features.

Lung adenocarcinoma (LUAD) is a predominant subtype of lung cancer. Although molecular classification of LUAD has been widely explored, proteomics-based subtyping of LUAD remains scarce. We proposed a subtyping method for LUAD based on the expression profiles of 500 proteins with the largest expression variability across LUAD. Furthermore, we comprehensively compared molecular and clinical features among the LUAD subtypes. Consensus clustering identified three subtypes of LUAD, namely MtE, DrE, and StE. We demonstrated this subtyping method to be reproducible by analyzing two independent LUAD cohorts. MtE was characterized by high enrichment of metabolic pathways, high EGFR mutation rate, low stemness, proliferation, invasion, metastasis and inflammation signatures, favorable prognosis; DrE was characterized by high enrichment of DNA repair pathways, high TP53 mutation rate, and high levels of genomic instability, stemness, proliferation, and intratumor heterogeneity (ITH); and StE was characterized by high enrichment of stroma-related pathways, high KRAS mutation rate, and low levels of genomic instability. The proteomics-based clustering analysis identified three LUAD subtypes with significantly different molecular and clinical properties. The novel subtyping method offers new perspectives on the cancer biology and holds promise in improving the clinical management of LUAD.

Peculiarities of neoplasms appeared after total body irradiation and homeostasis parameters in rats

Background: Tissue damage and disruption of metabolic processes as a result of total body irradiation (TBI) could lead to tumorigenesis. Methods: Female rats (25 of 32) were X-irradiated with a 6-Gy dose. On month 12±1 animals were sacrificed. The alkaline phosphatase (ALP), alanine aminotransferase (ALT), aspartate aminotransferase (AST), amylase, lactate dehydrogenase (LDH), Ca 2+, creatinine, glucose, phosphorus, urea, uric acid, total protein, pO2, pCO2, pH, and blood cell count were evaluated in blood. Tumors were examinated histologically. Results: On 12±1 months after TBI, 76% of rats had visually detected tumors, histologically classified as benign fibro adenomas. Metabolic, hematological changes versus healthy control indicated disturbances in the homeostasis system. The blood lactate level was typically higher in animals with tumors than without. The ratio of tumor weight to lactate (or LDH) level in blood was 1±0.5 in the 63% of rats and histological analysis revealed the signs of biphasic hyperplasia of glandular lobes and connective tissue stroma, associated with secretory and proliferative activities in tumor. In animals with high values of this ratio (≥1.5) neoplasms were represented by fibrous and glandular tissues presenting a predominant stromal fibrous component, associated with the prevalence of high proliferation in tumor. While in 20% of tumor bearing rats with the low ratio (<0.5) predominated an epithelial structure with homogeneous basophilic content in the glandular lumens, suggesting the domination of secretory activity in tumor. Conclusions: TBI promoted the alterations of hematological and biochemical parameters of homeostasis in rats and provoked the appearance of benign tumors one year after. The ratio of tumor mass to lactate (or LDH) level in blood seems to be an informative indicator of the histological particularities of tumors, suggesting the prevalence of proliferative or secretory activity, or the balance between them.

Clinical Significance of Carnitine in the Treatment of Cancer: From Traffic to the Regulation.

Metabolic reprogramming is a common hallmark of cancer cells. Cancer cells exhibit metabolic flexibility to maintain high proliferation and survival rates. In other words, adaptation of cellular demand is essential for tumorigenesis, since a diverse supply of nutrients is required to accommodate tumor growth and progression. Diversity of carbon substrates fueling cancer cells indicate metabolic heterogeneity, even in tumors sharing the same clinical diagnosis. In addition to the alteration of glucose and amino acid metabolism in cancer cells, there is evidence that cancer cells can alter lipid metabolism. Some tumors rely on fatty acid oxidation (FAO) as the primary energy source; hence, cancer cells overexpress the enzymes involved in FAO. Carnitine is an essential cofactor in the lipid metabolic pathways. It is crucial in facilitating the transport of long-chain fatty acids into the mitochondria for β-oxidation. This role and others played by carnitine, especially its antioxidant function in cellular processes, emphasize the fine regulation of carnitine traffic within tissues and subcellular compartments. The biological activity of carnitine is orchestrated by specific membrane transporters that mediate the transfer of carnitine and its derivatives across the cell membrane. The concerted function of carnitine transporters creates a collaborative network that is relevant to metabolic reprogramming in cancer cells. Here, the molecular mechanisms relevant to the role and expression of carnitine transporters are discussed, providing insights into cancer treatment.

The Use of Wheat Starch as Gelling Agent for In Vitro Proliferation of Blackberry (Rubus fruticosus L.) Cultivars and the Evaluation of Genetic Fidelity after Repeated Subcultures

Micropropagation has an important role in the large-scale production of blackberry plant material, given the high proliferation rates of this species. The aim of the present study was to evaluate the proliferative capacity of blackberry grown in vitro on wheat starch-gelled culture medium compared to classical agar-gelled medium and to assess the genetic fidelity between the proliferated shoots in starch-gelled culture medium and their mother plants. Six blackberry varieties (‘Čačanska Bestrna’, ‘Chester Thornless’, ‘Driscoll’s Victoria’, ‘Loch Ness’, ‘Polar’, and ‘Karaka Black’) were tested. For the in vitro shoots proliferation, Murashige and Skoog (MS) medium supplemented with 0.5 mg dm−3 6-benzyladenine (BA) was used. The conventional medium was gelled with 0.5% plant agar, and wheat starch was used as an alternative gelling agent in a concentration of 5%. The results showed that for all blackberry cultivars, the highest number of shoots/inoculum was obtained in wheat starch-gelled culture medium, with a maximum value of 54.42 ± 4.18 presented by ‘Karaka Black’. Considering the length of the proliferated shoots, all tested cultivars presented outstanding results on the culture medium gelled with 5% wheat starch. The highest values regarding shoots length were observed on the ‘Chester Thornless’ followed by ‘Čačanska Bestrna’, and ‘Loch Ness’ with values of 5.55 ± 0.04 cm, 5.46 ± 0.06 cm, and 5.37 ± 0.09 cm, respectively. The genetic uniformity of the micropropagated shoots in relation to their mother plants was confirmed by sequence-related amplified polymorphism (SRAP) and start codon targeted (SCoT) molecular markers.

Dynamic matrices with DNA-encoded viscoelasticity for cell and organoid culture

Three-dimensional cell and organoid cultures rely on the mechanical support of viscoelastic matrices. However, commonly used matrix materials lack control over key cell-instructive properties. Here we report on fully synthetic hydrogels based on DNA libraries that self-assemble with ultrahigh-molecular-weight polymers, forming a dynamic DNA-crosslinked matrix (DyNAtrix). DyNAtrix enables computationally predictable and systematic control over its viscoelasticity, thermodynamic and kinetic parameters by changing DNA sequence information. Adjustable heat activation allows homogeneous embedding of mammalian cells. Intriguingly, stress-relaxation times can be tuned over four orders of magnitude, recapitulating mechanical characteristics of living tissues. DyNAtrix is self-healing, printable, exhibits high stability, cyto- and haemocompatibility, and controllable degradation. DyNAtrix-based cultures of human mesenchymal stromal cells, pluripotent stem cells, canine kidney cysts and human trophoblast organoids show high viability, proliferation and morphogenesis. DyNAtrix thus represents a programmable and versatile precision matrix for advanced approaches to biomechanics, biophysics and tissue engineering.

CRISPR-Cas9 based knockout of S100A8 in mammary epithelial cells enhances cell proliferation and triggers oncogenic transformation via the PI3K-Akt pathway: Insights from a deep proteomic analysis

S100A8 is a calcium-binding protein with multiple functions, including being a chemoattractant for phagocytes and playing a key role in the inflammatory response. Its expression has been shown to influence epithelial-mesenchymal transition (EMT) and metastasis in colorectal cancer. However, the role of S100A8 in cell proliferation and differentiation remains unknown. In this study, we used the CRISPR-Cas9 system to knock out S100A8 in healthy mammary epithelial cells and investigated the resulting changes in proteome profiling and signaling pathways. Our results showed that S100A8 knockout led to an increase in cell proliferation and migration, reduced cell-cell adhesion, and increased apoptosis compared to wildtype cells. Proteomics data indicated that S100A8 significantly affects cell cycle progression, cell proliferation, and cell survival through the PI3K-Akt pathway. Furthermore, our findings suggest that S100A8 function is associated with Pten expression, a negative regulator of the PI3K-Akt pathway. These results indicate that S100A8 dysregulation in healthy cells can lead to altered cellular physiology and higher proliferation, similar to cancerous growth. Therefore, maintaining S100A8 expression is critical for preserving healthy cell physiology. This study provides novel insights into the role of S100A8 in cell proliferation and differentiation and its potential relevance to cancer biology. SignificanceThe study suggests that maintaining S100A8 expression is critical for preserving healthy cell physiology, and dysregulation of S100A8 in healthy cells can lead to altered cellular physiology and higher proliferation, similar to cancerous growth. Therefore, targeting the PI3K-Akt pathway or regulating Pten expression, a negative regulator of the PI3K-Akt pathway, may be potential strategies for cancer treatment by controlling S100A8 dysregulation. Additionally, S100A8 and S100A9 have been shown to promote metastasis of breast carcinoma by forming a metastatic milieu. However, the differential expression of S100A8 in tumors and its dual effects of antitumor and protumor make the relationship between S100A8 and tumors complicated. Currently, most research focuses on the function of S100A8 as a secretory protein in the microenvironment of tumors, and its function inside healthy cells without forming dimers remains unclear. Furthermore, the study provides insight into the role of S100A8 in cell proliferation and differentiation, which may have implications for other diseases beyond cancer. The functional role of S100A8 in normal mammary epithelial cells remains completely uncertain. Therefore, the objective of this study is to investigate the function of S100A8 on proliferation in mammary epithelial cells after its deletion and to elucidate the underlying proteins involved in downstream signaling. Our findings indicate that the deletion of S100A8 leads to excessive proliferation in normal mammary epithelial cells, reduces apoptosis, and affects cell-cell adhesion molecules required for cellular communication, resulting in a cancer-like phenotype.

Tumor-Targeted Nonablative Radiation Promotes Solid Tumor CAR T-cell Therapy Efficacy.

Infiltration of tumor by T cells is a prerequisite for successful immunotherapy of solid tumors. In this study, we investigate the influence of tumor-targeted radiation on chimeric antigen receptor (CAR) T-cell therapy tumor infiltration, accumulation, and efficacy in clinically relevant models of pleural mesothelioma and non-small cell lung cancers. We use a nonablative dose of tumor-targeted radiation prior to systemic administration of mesothelin-targeted CAR T cells to assess infiltration, proliferation, antitumor efficacy, and functional persistence of CAR T cells at primary and distant sites of tumor. A tumor-targeted, nonablative dose of radiation promotes early and high infiltration, proliferation, and functional persistence of CAR T cells. Tumor-targeted radiation promotes tumor-chemokine expression and chemokine-receptor expression in infiltrating T cells and results in a subpopulation of higher-intensity CAR-expressing T cells with high coexpression of chemokine receptors that further infiltrate distant sites of disease, enhancing CAR T-cell antitumor efficacy. Enhanced CAR T-cell efficacy is evident in models of both high-mesothelin-expressing mesothelioma and mixed-mesothelin-expressing lung cancer-two thoracic cancers for which radiotherapy is part of the standard of care. Our results strongly suggest that the use of tumor-targeted radiation prior to systemic administration of CAR T cells may substantially improve CAR T-cell therapy efficacy for solid tumors. Building on our observations, we describe a translational strategy of "sandwich" cell therapy for solid tumors that combines sequential metastatic site-targeted radiation and CAR T cells-a regional solution to overcome barriers to systemic delivery of CAR T cells.

Decoding the prognostic significance of integrator complex subunit 9 (INTS9) in glioma: links to TP53 mutations, E2F signaling, and inflammatory microenvironments

IntroductionGliomas, a type of brain neoplasm, are prevalent and often fatal. Molecular diagnostics have improved understanding, but treatment options are limited. This study investigates the role of INTS9 in processing small nuclear RNA (snRNA), which is crucial to generating mature messenger RNA (mRNA). We aim to employ advanced bioinformatics analyses with large-scale databases and conduct functional experiments to elucidate its potential role in glioma therapeutics.Materials and methodsWe collected genomic, proteomic, and Whole-Exon-Sequencing data from The Cancer Genome Atlas (TCGA) and Chinese Glioma Genome Atlas (CGGA) for bioinformatic analyses. Then, we validated INTS9 protein expression through immunohistochemistry and assessed its correlation with P53 and KI67 protein expression. Gene Set Enrichment Analysis (GSEA) was performed to identify altered signaling pathways, and functional experiments were conducted on three cell lines treated with siINTS9. Then, we also investigate the impacts of tumor heterogeneity on INTS9 expression by integrating single-cell sequencing, 12-cell state prediction, and CIBERSORT analyses. Finally, we also observed longitudinal changes in INTS9 using the Glioma Longitudinal Analysis (GLASS) dataset.ResultsOur findings showed increased INTS9 levels in tumor tissue compared to non-neoplastic components, correlating with high tumor grading and proliferation index. TP53 mutation was the most notable factor associated with upregulated INTS9, along with other potential contributors, such as combined chromosome 7 gain/10 loss, TERT promoter mutation, and increased Tumor Mutational Burden (TMB). In GSEA analyses, we also linked INTS9 with enhanced cell proliferation and inflammation signaling. Downregulating INTS9 impacted cellular proliferation and cell cycle regulation during the function validation. In the context of the 12 cell states, INTS9 correlated with tumor-stem and tumor-proliferative-stem cells. CIBERSORT analyses revealed increased INTS9 associated with increased macrophage M0 and M2 but depletion of monocytes. Longitudinally, we also noticed that the INTS9 expression declined during recurrence in IDH wildtype.ConclusionThis study assessed the role of INTS9 protein in glioma development and its potential as a therapeutic target. Results indicated elevated INTS9 levels were linked to increased proliferation capacity, higher tumor grading, and poorer prognosis, potentially resulting from TP53 mutations. This research highlights the potential of INTS9 as a promising target for glioma treatment.

Effects of Tricalcium Silicate Cement on Corneal Cell Proliferation and Its Relationship with Vascular Endothelial Growth Factor Level and Receptor Typing

This research analyzed the effects of tricalcium silicate (C3S) cement and hypoxia on proliferation of human corneal epithelial cells (HCEpiCs) and the levels of vascular endothelial growth factor (VEGF) and its receptors (VEGFRs). Nanoscale C3S was prepared using a combustion method and characterized using X-ray diffraction (XRD), scanning electron microscopy (SEM), and laser particle size (LPS) analyzer. HCEpiCs were cultured, and influences of C3S with changed concentrations on proliferation of (HCEpiCs were analyzed. The cells were treated with hypoxia or with low-concentration (0.5 mg/mL, LC-C3S), medium-concentration (5 mg/mL, MC-C3S), or high-concentration (50 mg/mL, HC-C3S) of C3S. Meanwhile, normal HCEpiCs were undertaken as controls (Ctrl group). Cell proliferation, apoptosis, and the expression of target genes were detected using CCK-8, Annexin V-FITC/PI, fluorescent quantitative polymerase chain reaction (fqPCR), and Western blotting (WB). The results suggested that nanoscale C3S had multiple morphologies and an average particle size (APS) of (231.5±8.3) nm. With increasing nanoscale C3S concentration, proliferation of HCEpiCs increased (P < 0.05), and the highest proliferation was visualized at 5 mg/mL. Based on the conditions in the Ctrl group, the hypoxia group exhibited a decreased proliferation rate (PR), an increased apoptosis rate (AR), downshifted VEGF, VEGFR-2, and VEGFR-3, and elevated VEGFR-1 (P < 0.05). Based on the hypoxia group, the LCC3S, MC-C3S, and HC-C3S groups presented increased cell PRs, decreased APs, upshifted VEGF, VEGFR-2, and VEGFR-3, and downregulated VEGFR-1 (P < 0.05). The MC-C3S group showed an increased cell PR, a decreased AP, upregulated VEGF, VEGFR-2, and VEGFR-3, and downregulated VEGFR-1 to the LC-C3S group (P < 0.05). Additionally, the HC-C3S group had a decreased cell PR, an increased AP, upregulated VEGF, VEGFR-2, and VEGFR-3, and a downshifted VEGFR-1 to the MC-C3S group (P < 0.05). Therefore, C3S promoted proliferation of HCEpiCs, upregulated VEGF, VEGFR-2, and VEGFR-3, and downregulated VEGFR-1.

Skip metastasis at the esophageal resection margin in radical gastrectomy: clinical characteristics of 30 cases

Objective: To summarize the clinical characteristics of patients with skip metastasis at esophageal resection margin during radical gastrectomy. Methods: This is a descriptive study of case series. Relevant data from 2006 to 2022 were collected from two major gastric cancer consultation and treatment centers: Nanjing Drum Tower Hospital and Jinling Hospital.Characteristics, surgical approach, number of dissected lymph nodes, immunohistochemical staining, and pathological staging were summarized and analyzed. The distribution of residual tumor cells at the esophageal margins was further analyzed at the cellular and tissue levels. Skip metastasis at the esophageal resection margin was defined as a negative esophageal margin with a positive margin in the cephalad donut. Results: Thirty (0.33%, 30/8972) eligible patients, 24 (80.0%) of whom were male, were identified in the two centers. The mean age was 63.9±11.0 years. Seventeen (56.7%) of these patients had papillary or tubular adenocarcinomas, including 13 (43.3%) poorly- and four (13.3%) moderately-differentiated tumors; four (13.3%) had signet-ring cell carcinomas; four (13.3%) mucinous adenocarcinomas; three (10.0%) mixed adenocarcinomas, including two with poorly-differentiated tubular adenocarcinomas mixed with signet-ring cell carcinoma and mucinous adenocarcinoma; and one had a poorly differentiated tubular adenocarcinoma mixed with signet-ring cell carcinoma. Two patients (6.7%) had other types of cancer, namely adenosquamous carcinoma in one patient and undifferentiated carcinoma in the other one. The predominant tumor sites were the lesser curvature (n=26, 86.7%) and the cardia (n=24, 80.0%). The mean tumor diameter was 6.6 cm, mean distance between tumor and esophageal resection margin was 1.5 cm, and proportions of tumor invasion into the dentate line, nerves, and vessels were 80.0% (24/30), 86.7%(26/30), and 93.3% (28/30), respectively. The mean number of lymph nodes resected was 20.4±8.9. The pathological stage was mainly T4 (n=18, 60.0%) and N3 (n=21, 70.0%), the median Ki67 was 52.7%, and the rates of positivity for HER2, EGFR, VEGFR, E-cadherin and PD-L1 were 40.0% (12/30), 46.7% (14/30), 80.0% (24/30), 86.7% (26/30) and 16.7% (5/30), respectively. At the cellular level, cancer cells were mainly distributed in small focal areas, as cell masses, or as tumor thrombi; large numbers of widely distributed atypic cells were seldom observed. At the tissue level, cancer cells were located in the mucosal layer in seven patients (23.3%), in the submucosal layer in 18 (60.0%), and in the muscular layer in five (16.7%); no cancer cells were identified in the outer membrane. Five of the seven tumors were located in the lamina propria, two in the muscularis mucosae, and none in the mucosal epithelium. Conclusion: Patients with skip metastasis at the esophageal resection margin at radical gastrectomy have unfavorable tumor biology and a high proliferation index, are at a late pathological stage, and the residual cancer is mostly located in the submucosa.

Leishmania major drives host phagocyte death and cell-to-cell transfer depending on intracellular pathogen proliferation rate.

The virulence of intracellular pathogens relies largely on the ability to survive and replicate within phagocytes but also on release and transfer into new host cells. Such cell-to-cell transfer could represent a target for counteracting microbial pathogenesis. However, our understanding of the underlying cellular and molecular processes remains woefully insufficient. Using intravital 2-photon microscopy of caspase-3 activation in the Leishmania major-infected (L. major-infected) live skin, we showed increased apoptosis in cells infected by the parasite. Also, transfer of the parasite to new host cells occurred directly without a detectable extracellular state and was associated with concomitant uptake of cellular material from the original host cell. These in vivo findings were fully recapitulated in infections of isolated human phagocytes. Furthermore, we observed that high pathogen proliferation increased cell death in infected cells, and long-term residency within an infected host cell was only possible for slowly proliferating parasites. Our results therefore suggest that L. major drives its own dissemination to new phagocytes by inducing host cell death in a proliferation-dependent manner.

Piperine suppresses inflammatory fibroblast-like synoviocytes derived from rheumatoid arthritis patients Via NF-κB inhibition

Rheumatoid Arthritis (RA) is a common autoimmune disease recognized by hyperplasia of synoviocytes and chronic joint inflammation. Activation of fibroblast-like synoviocytes (FLSs) is one of the main features of RA which can trigger inflammation leading to articular cartilage and joint destruction. Aberrant activation of NF-κB signaling cascade was found to be responsible for the high proliferation and defective apoptosis of FLSs and subsequent inflammation in RA. Piperine is a principal constituent of piper species frequently used as antitumor and anti-inflammatory natural compound. In this study we aimed to assess the anti-inflammatory effect of piperine on RA-FLS through NF-κB inhibition.FLSs were isolated from 68 RA patients and 30 healthy controls and were exposed to piperine. The main assays were MTT assay, flow cytometric analysis, PI staining, reverse transcription-PCR (RT-PCR), and ELISA.Results showed that piperine can induce the apoptosis and reduce the proliferation of RA-FLSs in vitro. Moreover, piperine directly reduced the phosphorylation of NF-kB and the expression of NF-κB target genes related to RA-FLSs proliferation (c-Myc and Cycline D1), apoptosis inhibition (Bcl2 and Bcl-xl) and inflammation (COX2, IL-1β, TNF-α,IL-6, CCL5 and CXCL10) while increasing the expression of apoptosis related ones (Bax) in vitro. Piperine also reduced the protein levels of cytokines and chemokines secreted by FLSs as a result of NF-κB inhibition.In conclusion, our results provide evidence for the anti-inflammatory capacity of piperine through inhibition of NF-κB pathway in FLSs proposing this compound as a suitable alternative for chemical treatment of RA.