Tissue Samples Research Articles

Ecytonucleospora hepatopenaei (EHP) disease prevalence and mortality in Litopenaeus vannamei: a comparative study from Eastern India shrimp farms

Ecytonucleospora hepatopenaei (EHP), a microsporidian parasite first named and characterized from the Penaeus monodon (black or giant tiger shrimp), causes growth retardation and poses a significant threat to shrimp farming. We observed shrimp farms associated with disease conditions during our fish disease surveillance and health management program in West Bengal, India. Shrimp exhibited growth retardation and increased size variability, particularly in advanced stages, exhibiting soft shells, lethargy, reduced feeding and empty midguts. Floating white feces were observed on the surface of the pond water. Suspecting a microbial infection, the shrimp samples were collected and aseptically brought to the ICAR-CIFRI laboratory for molecular confirmation. A nested PCR was used to screen shrimp tissue, feces, feed and environmental samples for the possible presence of hepatopancreatic microsporidiosis caused by Ecytonucleospora hepatopenaei. The results confirmed that the shrimp samples were positive for EHP. Histopathological investigation revealed mature spores in the HP tubule lumen and epithelial cells along with necrotic tubule in the symptomatic group. Further, the transcription analysis revealed that ProPO, Hsp70 and α2-macroglobulin genes were significantly upregulated, while decreased expression of LGBP, PXN and Integrin ß was observed in shrimp infected with Hepatopancreatic microsporidiosis. Furthermore, compared with the healthy group, significant intestinal bacteria changes were observed in the EHP-infected group. The in vivo survival assay, using crustacean animal model Artemia franciscana, suggests that symptomatic shrimp gut samples harbour pathogenic Vibrio parahaemolyticus, V. harveyi and V. campbellii. These results significantly advance our understanding of the molecular and ecological aspects of EHP pathobiology.

Effect of age and sex differences on the abundances of neuronal, glial, and endothelial cells in non-diseased brain tissue

Neuroinflammatory and neurodegenerative diseases are influenced by the complex interplay of different cell types within the brain, and understanding the proportions and dynamics of neuronal, glial, and endothelial cells is crucial for deciphering the mechanisms of these diseases. Certain risk factors, such as age and sex differences, are thought to play a significant role in the susceptibility, progression, and response to neurological disease. Therefore, investigation of age- and sex-related differences in cell type proportions is needed to elucidate the biological basis of these diseases. Advances in sequencing technology have enabled large-scale transcriptomic studies, such as the Genotype-Tissue Expression (GTEx) project, providing valuable resources for investigating the cellular landscape of the human brain. In this analysis, we used brain sample data from the GTEx project, comprising 1646 samples with an age range of 20–70 years. The relative abundance of excitatory and inhibitory neurons, astrocytes, oligodendrocytes, microglia, and endothelial cells was estimated from the RNA sequencing data using a deconvolution-based analysis. Spearman correlation analysis between the individuals’ calendar ages and cell type proportions revealed a statistically significant decrease in the proportion of neurons with increasing age. In contrast, the proportions of astrocytes and endothelial cells showed a significant increase. Furthermore, endothelial cells exhibited the strongest correlation coefficient, positively associating with age. In addition, the findings indicate sex-based differences in age-related changes to cell type proportions. An age-associated decrease in neuronal proportions was only observed in male donors, while no significant change was found in females. Additionally, an age-associated increase in astrocyte proportions was exclusively seen in males, whereas only females exhibited a significant increase in microglia proportions. Furthermore, we identified sex-based differences in baseline cell type proportions. Male originating samples exhibited higher proportions of excitatory neurons, while female samples showed higher proportions of microglia and endothelial cells. Our results show that both age and sex affect the proportions of cell types in non-diseased brain tissue samples. These findings contribute to our understanding of the effects of age and sex differences on the cellular composition of the brain and shed light on the potential roles of age and sex in neurological diseases.

Development of a novel diagnostic model to monitor the progression of metabolic dysfunction-associated steatotic liver disease to hepatocellular carcinoma in females.

The onset of metabolic dysfunction-associated steatotic liver disease-associated hepatocellular carcinoma (MASLD-HCC) is insidious and exhibits sex-specific variations. Effective methods for monitoring MASLD-HCC progression in females have not yet been developed. Transcriptomic data of female liver tissue samples were obtained from multiple public databases. Differentially expressed genes (DEGs) in MASLD-HCC were identified using differential expression and robust rank aggregation analyses. Diagnostic prediction models for MASLD (DP.MASLD) and HCC (DP.HCC) were developed and validated using elastic net analysis, and diagnostic performance was evaluated using receiver operating characteristic (ROC) curve analysis. Bioinformatics was used to assess the pathogenesis of MASLD-HCC. Seven overlapping DEGs were identified in female patients with MASLD and HCC: AKR1B10, CLEC1B, CYP2C19, FREM2, MT1H, NRG1, and THBS1). The area under the ROC curve (AUC) values for the training and validation groups of the DP.MASLD model were 0.864 and 0.782, 0.932 and 1.000, and 0.920 and 0.969 when differentiating between the steatosis and normal liver, steatohepatitis and steatosis, and steatohepatitis and normal liver groups, respectively. The AUCs for DP.HCC were 0.980 and 0.997 in the training and validation groups, respectively. The oncogenesis of female MASLD-HCC is associated with molecular pathways, including cytochrome P450-associated drug metabolism, tyrosine metabolism, fatty acid degradation, focal adhesion, extracellular matrix receptor interactions, and protein digestion and absorption. A novel and effective method to quantitatively assess the risk of MASLD-HCC progression in female patients was developed, and this method will aid in the generation of precise diagnostic, preventive, and therapeutic strategies.

MiR-484 as an "OncomiR" in Breast Cancer Promotes Tumorigenesis by Suppressing Apoptosis Genes.

Breast cancer (BC) is one of the most common causes of death among females. Cancer cells escape from apoptosis, causing the cells to proliferate uncontrollably. MicroRNAs (miRNAs) are known to regulate apoptosis in cancer cells. This study aimed to determine the change in miR-484 in different BC cells and its relationship with the apoptosis pathway. In the study, tumor and healthy tissue samples adjacent to the tumor were collected from 42 patients (6 benign, 36 malignant). Tissue samples were classified according to tumor type, tumor histological grade, proliferation index, and molecular subtypes. Gene expression levels were determined by quantitative real-time polymerase chain reaction (qRT-PCR), and protein levels were determined using the Western Blot method. The results were analyzed using the delta-delta Ct method. Findings showed that miR-484 expression levels were higher in malignant tumors than in benign tumors, and higher in tumor tissues than healthy tissues. Additionally, it was determined that as Ki-67 levels and histological grade and aggressiveness increased, miR-484 expression levels also increased. In tumor tissue compared with healthy adjacent tissue, there was an increase in BCL2 expression and a decrease in Casp3 and Casp9 expression. Therefore, a positive correlation was found between miR-484 expression and BCL2, and a negative correlation was found between CASP3 and CASP9 expression. Our results show that miR-484 may play a roll as an onco-miR in BC. Increased miR-484 and BCL2, and decreased Casp3, in breast tumor tissues suggest that Casp9 expression may increase uncontrolled cell proliferation by suppressing apoptosis in BC cells and may contribute to tumor progression.

The Impact of Argan Oil and Polyhexanide on Healing Wounds with Tissue Loss Infected by Staphylococcus aureus in Mice

Background: One of the most important complications of the wound is tissue infection. One of the most common causes of tissue infections is microorganisms such as Staphylococcus. This research investigates the impact of argan oil and polyhexanide on the healing process in wounds with tissue loss infected with Staphylococcus aureus (S. aureus). Methods: The study involved 45 male mice, with 10 mm skin incisions made on their backs under general anaesthesia to create wounds. After 24 hours, an S. aureus (ATCC) suspension was applied to the scars. Wound cultures were collected from each mouse 48 hours later and this was repeated every 48 hours until cultures confirmed S. aureus infection. Treatment commenced once S. aureus growth was detected. The mice were randomly assigned to three groups: a control group (wound created, but no treatment applied), an argan group (1 mL of argan oil administered to the wound via syringe), and a polyhexanide group (1 mL of polyhexanide administered similarly). The wound diameters and clinical symptoms were monitored daily. On the 7th and 14th days, tissue samples were taken from the sacrificed mice for histopathological examination. Result: Based on clinical and histopathological observations, both argan oil and polyhexanide were found to be effective in treating S. aureus-infected wounds with tissue loss in mice. However, argan oil demonstrated a superior therapeutic potential compared to polyhexanide.

The Balance of Ketoacids α-Ketoglutarate and α-Ketoglutaramate Reflects the Degree of the Development of Hepatoencephalopathy in Rats

Hepatoencephalopathy (HE) is a liver disease that can lead to brain pathology and the impairment of human cognitive abilities. The objective assessment of HE disease severity is difficult due to the lack of reliable diagnostic markers. This paper examines the background to the emergence of HE markers and provides a brief overview of research results indicating the diagnostic value of potential markers isolated from a wide range of metabolites analyzed. It has been suggested that metabolites of the glutamate–glutamine (Glu-Gln) cycle, α-ketoglutarate (αKG), and α-ketoglutaramate (αKGM) can act as such markers of HE. The informative value of these markers was revealed during a comparative analysis of the distribution of αKG and αKGM in samples of the blood plasma and tissues (liver, kidneys, and brain) of rats exposed to the strong hepatotoxin thioacetamide (TAA). A comparative analysis of the balance of αKG and αKGM, as well as their ratio (αKG/αKGM) in the examined samples of blood plasma and animal tissues in these models, revealed their diagnostic value for assessing the severity of HE and/or monitoring the recovery process.

Effect of pre-farrowing hygiene routine (sub-standard vs. optimal) and creep feeding regime (dry pelleted starter diet vs. liquid mixture of milk replacer and starter diet) on post-weaning intestinal parameters and growth to slaughter in pigs.

The objective was to evaluate the effect of providing dry pelleted starter diet (DPS) or a liquid mixture of milk replacer and starter diet (LMR+S) to suckling pigs housed in farrowing pens of sub-standard or optimal hygiene conditions on pig growth to slaughter, and post-weaning (PW) intestinal parameters. On day (d) 107 of gestation, 87 sows were randomly allocated to one of four treatments in a 2×2 factorial arrangement. The factors were creep feeding (DPS or LMR+S) and pre-farrowing hygiene routine (SUB-STANDARD or OPTIMAL). Pigs were provided with DPS (manually) from d11 to weaning (at d28±1.2 of age) or LMR+S using an automatic liquid feeding system from d4 to weaning. The SUB-STANDARD hygiene routine (pens washed and dried for ~18h, sows not washed or disinfected) and the OPTIMAL hygiene routine (pens pre-soaked, detergent applied, washed, dried for 3 days, chlorocresol-based disinfectant applied, dried for 3 more days and sows washed and disinfected with Virkon) were used to obtain SUB-STANDARD or OPTIMAL hygiene conditions, respectively in farrowing rooms prior to entry of sows. Microbiome analysis was performed on fecal samples from 8 focal pigs/treatment, before weaning and at d21 and d114 PW. On d4 PW, 10 pigs/treatment were euthanized to collect intestinal tissue and digesta samples for histological, enzyme activity and microbiome analysis. Feeding LMR+S to pigs born into the OPTIMAL hygiene increased total dry matter intake compared to all of the other groups (P ≤ 0.05) and increased weaning weight compared to DPS feeding under both OPTIMAL and SUB-STANDARD hygiene conditions (P ≤ 0.05). Pigs from OPTIMAL farrowing pens had lower clinical cases of disease, diarrhoea prevalence and were slaughtered 3.8 days earlier than those from SUB-STANDARD farrowing pens (P ≤ 0.05). Suckling piglet mortality was reduced with LMR+S (P ≤ 0.05). On d4 PW, jejunal and ileal villus height were increased by OPTIMAL hygiene and ileal sucrase activity was increased by LMR+S (P ≤ 0.05). On d4 PW, LMR+S-fed pigs from OPTIMAL farrowing pens had lower relative abundance of Clostridium_P in the jejunum. In conclusion, the OPTIMAL hygiene routine increased pre-weaning LMR+S feed intake, reduced clinical cases of disease, improved intestinal structure and reduced the weaning to slaughter duration, while LMR+S feeding increased weaning weight, intestinal maturity and reduced pre-weaning mortality.

The impact of a short-term high-fat diet on coagulation function in a mouse model and its role in exacerbating concanavalin A-induced liver injury

BackgroundRecently, the number of patients with metabolic dysfunction-associated steatotic liver disease (MASLD) and its more advanced condition, metabolic dysfunction-associated steatohepatitis (MASH), has been increasing. These patients are at a higher risk of cardiovascular events and thromboembolism. However, the direct impact of high-fat diet (HFD), a cause of MASLD, on liver coagulation function is not well understood. Previously, we demonstrated that a short-term, 4-day intake of a HFD exacerbates concanavalin A (Con A)-induced acute liver injury in mice by promoting coagulation and inflammation. This model demonstrates that the liver exposed to a short-term HFD is vulnerable even before disease onset. In this study, using this model, we elucidated the detailed mechanisms by which short-term HFD intake promotes coagulation, considering primary and secondary hemostasis.MethodsC57BL/6 mice normally fed a normal diet (ND) were subjected to a HFD for 4 days. Liver tissue and blood samples were collected before and 4 and 24 h after Con A administration. Histological analysis, flow cytometry for platelet analysis, and blood coagulation tests related to secondary hemostasis were performed.ResultsEven with short-term consumption of a HFD alone, platelet and fibrinogen levels increased in the peripheral blood and liver. Additionally, when Con A was administered to mice on a short-term HFD, an increase in P-selectin expression was observed in the liver, with no upregulation in peripheral blood platelets. Furthermore, in mice subjected to a short-term HFD and treated with Con A, prolonged prothrombin time (PT) and activated partial thromboplastin time (APTT) were observed.ConclusionsConsuming a HFD in short-term can enhance primary and secondary hemostasis, thereby increasing the risk of thrombosis. These conditions are presumed to be a risk factor that exacerbates Con A-induced liver injury. The findings provide insight into early intervention strategies for chronic liver diseases, such as MASLD and MASH.

Challenges in Implementing Comprehensive Precision Medicine Screening for Ovarian Cancer

Precision medicine has revolutionised targeted cancer treatments; however, its implementation in ovarian cancer remains challenging. Diverse tumour biology and extensive heterogeneity in ovarian cancer can limit the translatability of genetic profiling and contribute to a lack of biomarkers of treatment response. This review addresses the barriers in precision medicine for ovarian cancer, including obtaining adequate and representative tissue samples for analysis, developing functional and standardised screening methods, and navigating data infrastructure and management. Ethical concerns related to patient consent, data privacy and health equity are also explored. We highlight the socio-economic complexities for precision medicine and propose strategies to overcome these challenges with an emphasis on accessibility and education amongst patients and health professionals and the development of regulatory frameworks to support clinical integration. Interdisciplinary collaboration is essential to drive progress in precision medicine to improve disease management and ovarian cancer patient outcomes.

The extracellular CIRP as a predictive marker for the endothelial dysfunction in chronic obstructive pulmonary disease combined with pulmonary hypertension

BackgroundPulmonary hypertension (PH) is a serious complication of chronic obstructive pulmonary disease (COPD), distinguished by pulmonary endothelial dysfunction. The extracellular cold-inducible RNA-binding protein (eCIRP) is a damage-associated molecular pattern (DAMP) that triggers inflammation and causes vascular endothelial dysfunction in COPD-PH.MethodsThe expression levels of CIRP were compared in peripheral lung tissues among 40 individuals. Moreover, A prospective analysis was conducted on serum levels of eCIRP, interleukin (IL) 1β, IL-33, endothelin-1 (ET-1), and nitric oxide (NO) in 150 COPD patients and 50 healthy control individuals at Jiangsu Taizhou Peoples Hospital. The study aimed to compare these serum levels and correlations among COPD-PH group, COPD non-PH group and the normal group.ResultsWe found higher CIRP levels in COPD-PH compared to COPD non-PH and the normal in lung tissue samples. A prospective analysis showed higher serum levels of eCIRP, IL-1β, IL-33, and ET 1 in COPD-PH, while a noticeable reduction in NO levels. There exists a correlation between the severity of COPD-PH and elevated levels of eCIRP, proinflammatory cytokines like IL-1β and IL-33, along with indicators of endothelial dysfunction like endothelin-1 ET-1 and NO. Moreover, the serum eCIRP level demonstrated a notable positive correlation with the levels of IL-1β, IL-33, PCT, and ET-1, while displaying a negative correlation with NO and Peripheral Oxygen Saturation (SpO2). Moreover, the serum eCIRP level demonstrated a notable positive correlation with the levels of IL-1β, IL-33, PCT, and ET-1, while displaying a negative correlation with NO and SpO2. Moreover, an assessment of independent risk factors for COPD-PH with ROC curve analysis, gauged the predictive value of serum eCIRP, IL-1β, IL-33, ET-1, and NO levels in diagnosing COPD-PH. Elevated eCIRP, IL-33, and ET-1 levels significantly correlated with COPD-PH, highlighting eCIRP’s strong predictive value for this condition.ConclusioneCIRP levels could serve as a valuable biomarker for predicting endothelial dysfunction in COPD-PH.

LADS: a powerful vaccine platform for cancer immunotherapy and prevention

BackgroundThe intracellular bacterium Listeria monocytogenes is an attractive vector for cancer immunotherapy as it can effectively deliver tumor antigens to antigen-presenting cells, leading to a robust antitumor response.ResultsIn this study, we developed a novel vaccine platform called Listeria-based Live Attenuated Double Substitution (LADS), which involves introducing two amino acid substitutions (N478AV479A) into the virulence factor listeriolysin O (LLO). LADS is a safe vaccine platform, with an attenuation of nearly 7000-fold, while retaining complete immunogenicity due to the absence of deletion of any virulence factors. We developed two LADS-based vaccines, LADS-E7 and LADS-AH1, which deliver the human papillomavirus (HPV) type 16 E7 oncoprotein and murine colon carcinoma immunodominant antigen AH1, respectively. Treatment with LADS-E7 or LADS-AH1 significantly inhibited and regressed established tumors, while also dramatically increasing the populations of tumor-infiltrated antigen-specific CD8+ T cells. RNA-sequencing analysis of tumor tissue samples revealed that LADS-E7 altered the expression of genes related to the immune response. Moreover, intratumoral injection of LADS-based vaccines induced strong antitumor responses, generating systemic antitumor responses to control distant tumor growth. Encouragingly, LADS-E7 or LADS-AH1 immunization effectively prevented tumor formation and growth.ConclusionsOur findings demonstrate that LADS-based vaccines represent a more powerful platform for the development of immunotherapeutic and preventive vaccines against cancers and infectious diseases.

Analysis of Fibroblast Growth Factor 2 Impact and Mechanism on Broncho-Pulmonary Dysplasia.

Epithelial-mesenchymal transition (EMT) of alveolar epithelial cells is an important mechanism for the onset and development of broncho-pulmonary dysplasia (BPD).The role of FGF-2 in BPD is currently unclear. The aim of our study is to investigate the expression of FGF-2 in lung tissue of BPD mice, to further clarify the effect of FGF-2 on EMT in alveolar epithelial cells and to actively search for possible signaling pathways. The BPD model was induced by exposure to hyperoxia. Lung tissue samples were collected and Hematoxylin and eosin (HE) staining was used to determine the modelling effect. Quantitative Real-time Polymerase Chain Reaction (QRT-PCR), immunohistochemistry were used to detect FGF-2 expression in BPD mice. To further investigate the effect of FGF-2 supplementation and deficiency on EMT in alveolar epithelial cells, A549 cells were cryopreserved, resuspended, cultured and passaged. Transforming growth factor-β1 (TGFβ1) was used to induce EMT. FGF-2 small interfering RNA fragments were synthesised and screened. Fbroblast growth factor receptor1 (FGFR1) expression was inhibited by BGJ398. (3-(4, 5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H tetrazolium) (MTS) assay was used to detect the effect of FGF-2 and Infigratinib (BGJ398) on cell proliferation. We used qRT-PCR and Western blot to detect the expression of epithelial cell markers, mesenchymal cell markers and EMT-related signaling pathway proteins. Our results showed that the successful established hyperoxia mice model were characteristic by BPD. Hyperoxia decreased FGF-2 on day 4, upregulated FGF-2 on day 21, which resulted in EMT. In vitro, we found that FGF-2 alone increased the expression of mesenchymal markers, decreased the expression of epithelial markers and activated Phosphatidylinositol 3-kinase/Protein kinase B (PI3K/AKT), Small mother against decapentaplegic (Smad), mitogen-activated protein kinase (P38) and extracellular signal-regulated kinase (ERK) signaling pathways. FGF-2 could not reverse but synergistically promote TGF-β1-induced EMT of alveolar epithelial cells. Silencing FGF-2 increased the expression of epithelial marker E-cadherin, inhibited the PI3K/AKT, Smad, and P38 signaling pathways activated by TGF-β1, but activated ERK signaling. FGF-2 receptor inhibitor BGJ398 reversed TGF-β1-induced EMT, decreased the expression of FGFR1, and inhibited ERK signaling pathway activation. FGF2 was closely associated with EMT in BPD mice. Both high and low levels of FGF2 promoted EMT in A549. The FGF-2 receptor inhibitor BGJ398 reversed TGF-β1-induced EMT in A549 by inhibiting the FGFR1/P-ERK signaling pathway.

Immunohistochemical Evaluation of Basal and Luminal Markers in Bladder Cancer: A Study from a Single Institution

Background: Bladder cancer (BC) presents significant molecular diversity, which affects both prognosis and treatment results. Immunohistochemistry (IHC) facilitates the identification of molecular subtypes and their relationships with clinicopathological features. Methods: We performed an IHC analysis on tissue samples from 107 BC patients, evaluating the expression of markers GATA3, CD44, CK5/6, and CK20. We applied two methods to classify the tumor samples into basal and luminal subtypes. The relationships between these marker expressions, molecular subtypes, clinicopathological characteristics, and TILs were explored. Results: Most samples showed the expression of GATA3 and CD44, with notable correlations found between CD44 and CK5/6 as well as GATA3 and CK20. CD44 and CD20 expression were linked to a poorer prognosis. Additionally, the luminal and basal subtypes had distinct TIL patterns, which influenced overall survival. A poor prognosis was associated with the basal subtype with low TIL infiltration and the luminal subtype with high TIL infiltration. Conclusions: Our study clarifies the molecular characteristics of BC, underlining the prognostic importance of CD44 expression and the role of TILs in influencing subtype-specific outcomes. IHC proves valuable in subtype identification and supports personalized treatment strategies.

Molecular genetic characterization of porcine reproductive and respiratory syndrome virus outbreak in Assam, India and neighbouring regions.

Porcine reproductive and respiratory syndrome (PRRS) is a significant swine disease with no effective vaccine due to high viral mutation rates. This study investigates a natural PRRS outbreak through molecular, pathological, and serological analyses. Nineteen affected pigs were clinically examined, and 10 underwent post-mortem examination. PRRS virus (PRRSV) presence was confirmed in all tissue samples by RT-PCR targeting open reading frame (ORF) 5 and ORF7 genes. Clinical signs, especially in boars and sows, included fever, appetite loss, movement reluctance, erythematous skin patches, vomiting, and abortions in sows. Post-mortem findings highlighted lung consolidation, severe lymph node enlargement, interstitial pneumonia with mononuclear cells, macrophage accumulation and necrotic cells in alveolar spaces. Multifocal myocarditis, lymphoid follicular degeneration, and follicular necrosis were observed in the tonsil, spleen, and lymph nodes. PRRSV-specific antibodies were detected in 32.75% of serum samples, confirming the outbreak. Phylogenetic analysis of the PRRSV-ORF5 and ORF7 genes revealed a close genetic relationship between the outbreak samples from Assam and recent outbreaks in Idukki, Kerala, India (2018), and neighbouring country China, indicating the circulation of Genotype 2 virus in Assam. However, the sequences showed some differences from the isolates of Mizoram, India. In conclusion, this study provides molecular and pathological evidence of a PRRSV outbreak, confirms the presence of PRRSV-specific antibodies and viral RNA, and shed light on the virus's genetic characteristics in India.

EUS-guided fine-needle biopsy versus fine-needle aspiration for histopathological evidence for type 1 autoimmune pancreatitis: A single-center retrospective study in China

ABSTRACT Background and Objectives EUS is recommended for guiding pancreatic tissue acquisition in suspected autoimmune pancreatitis (AIP) cases. However, there is a lack of comparative research on the effectiveness between EUS-guided fine-needle aspiration (EUS-FNA) and EUS-guided fine-needle biopsy (EUS-FNB) for diagnosing AIP in China. This study aimed to evaluate the diagnostic accuracy of EUS-guided tissue acquisition (EUS-TA) specifically for type 1 AIP. Methods Between 2010 and 2023, individuals with AIP who received EUS-TA at Changhai Hospital were included in the study. Results A total of 173 patients diagnosed with AIP who underwent EUS-TA were included in the final analysis. Of these, 104 patients (60.1%) received EUS-FNA, and 69 patients (39.9%) underwent EUS-FNB. Sufficient pancreatic tissue samples (>5 cells/high-power field) were obtained in 164 of 173 patients (94.8%), with success rates of 94.2% for EUS-FNA and 95.7% for EUS-FNB (P > 0.05). EUS-FNB exhibited higher rates of reliable level 1 histopathological findings (40.9% vs. 16.3%, P < 0.001) and reliable level 2 histopathological findings (33.3% vs. 12.2%, P < 0.001) compared with EUS-FNA. Furthermore, a higher occurrence of IgG4-positive plasma cell infiltration (>10 cells/high-power field) was observed with EUS-FNB compared with EUS-FNA (74.2% vs. 27.9%, P < 0.001). The multivariate logistic analysis also revealed that EUS-FNA was less effective in obtaining reliable evidence compared with EUS-FNB, as evident in both level 2 (P = 0.002; odds ratio, 0.21; 95% confidence interval, 0.08–0.56) and level 1 (P = 0.001; odds ratio, 0.19; 95% confidence interval, 0.08–0.49) histopathological evidence. Conclusions EUS-FNB demonstrates higher rates of level 1 and level 2 histopathological findings, as well as more abundant IgG4-positive plasma cell infiltration, compared with EUS-FNA.

Targeting liver cancer stem cells: the prognostic significance of MRPL17 in immunotherapy response

BackgroundLiver hepatocellular carcinoma (LIHC) ranks as the foremost cause of cancer-related deaths worldwide, and its early detection poses considerable challenges. Current prognostic indicators, including alpha-fetoprotein, have notable limitations in their clinical utility, thereby underscoring the necessity for discovering new biomarkers to improve early diagnosis and enable personalized treatment options.MethodThis investigation employed single-cell analysis techniques to identify stem cell-associated genes and assess their prognostic significance for LIHC patients, as well as the efficacy of immunotherapy, utilizing nonnegative matrix factorization (NMF) cluster analysis. A diagnostic model for LIHC was developed and validated through multiple datasets and various machine learning clustering methods. The XGBOOST algorithm identified MRPL17 as the most significant prognostic gene among those associated with stem cells. Additionally, the research explores the relationship between MRPL17 expression and immune cell infiltration. Immunofluorescence staining of LIHC tissue samples was conducted to evaluate the expression and prognostic value of MRPL17, as well as its correlation with KI67.ResultsThrough single-cell analysis, this study identified 14 essential stem cell-related genes, highlighting their significance in the diagnosis, prognostication, and potential treatment strategies for LIHC patients. Various machine learning algorithms indicated that MRPL17 is particularly associated with patient prognosis and responses to immunotherapy. Furthermore, experimental results demonstrate that MRPL17 is upregulated in LIHC and correlates with poor prognosis, as well as positively correlating with KI67.ConclusionCancer stem cells are pivotal in the mechanisms of immune evasion within the tumor microenvironment and have a substantial impact on treatment results. This study experimentally validated MRPL17 as a promising prognostic biomarker, emphasizing the need to target liver cancer stem cells to improve patient prognosis and enhance treatment effectiveness.

Impact of T Cell Exhaustion and Stroma Senescence on Tumor Cell Biology and Clinical Outcome of Head and Neck Squamous Cell Carcinomas

Head and neck squamous cell carcinomas (HNSCC) have an overall poor prognosis, especially in locally advanced and metastatic stages. In most cases, multimodal therapeutic approaches are required and show only limited cure rates with a high risk of tumor recurrence. Anti-PD-1 antibody treatment was recently approved for recurrent and metastatic cases but to date, response rates remain lower than 25%. Therefore, the investigation of the immunological tumor microenvironment and the identification of novel immunotherapeutic targets in HNSCC is of paramount importance. In our study, we used tissue samples of n = 116 HNSCC patients for the immunohistochemical detection of the intratumoral and peritumoral expression of T cell exhaustion markers (PD-1, LAG-3, TIM-3) on tumor infiltration leukocytes (TIL), as well as the expression level of stromal senescence markers (IL-8, MMP-3) on tumor-associated fibroblasts. The clinical parameter of the vitamin D serum status as well as the histopathological HPV infection status of the tumor was correlated with the expression rates of the biomarkers and the overall patient survival. An increased peritumoral and intratumoral expression of the biomarkers PD-1 and TIM-3 significantly correlated with improved overall patient survival. A high peritumoral expression of LAG-3 correlated with better overall survival. A positive HPV tumor status correlated with a significantly elevated expression of PD-1 and TIM-3. Biomarkers of stromal senescence showed no influence on the patient outcome. However, the vitamin D serum status showed no influence on patient outcomes or biomarker expressions. Our study identified PD-1, LAG-3, and TIM-3 as promising targets of a therapeutic strategy targeting the tumor microenvironment in HNSCC, particularly among HPV-positive patients, where a higher expression of these checkpoints correlated with an improved overall survival. These findings support the potential of antibodies targeting these immune checkpoints to enhance treatment efficacy, especially in the context of bispecific targeting.

Biophysical and biochemical study exploring resveratrol protective potential against gamma irradiation effects

BackgroundGamma irradiation causes oxidative stress and disturbs the physiological balance in organisms. Natural antioxidants, like resveratrol (Res), can buffer these effects. The study's goal is to find out whether Res can help reverse changes caused by gamma irradiation in male Wistar rats' total antioxidant capacity (TAC), antioxidant enzyme activity, electrolyte levels, and hemoglobin structure.MethodsTwenty-four rats were divided into four groups: control, Res-treated, irradiated, and Res + irradiated. Blood and tissue samples were collected on the day 16th of the experiment. The levels of antioxidants, catalase (CAT) activity, glutathione (GSH), and glutathione peroxidase (GPx) were estimated along with electrolyte level measurements. UV–FTIR and dielectric spectroscopy were used to assess conformational changes in hemoglobin.ResultsGamma irradiation decreased antioxidant levels and increased markers of oxidative stress. Resveratrol treatment increased the antioxidant capacity and levels of nitric oxide and decreased malondialdehyde levels. Further, Res protected the structure of hemoglobin and decreased the radiation-induced damage.ConclusionResveratrol shows potential as a protective agent against gamma irradiation-induced oxidative stress by enhancing antioxidant defenses and restoring hemoglobin structure.

Parotid and Sublingual Salivary gland as an Indicator of Antioxidant Efficacy in Experimentally Induced Diabetic Rats

The parotid and sublingual glands were considered the most appropriate tissues to study the effects of diabetes mellitus among the major salivary glands. Globally, diabetes mellitus is a major healthcare concern. It affects bodily tissues and organs in a variety of pathological ways, particularly the major salivary glands. Thirty adult, healthy albino rats were divided into three equal groups of ten animals each. Group I was a control group that did not receive any treatment, group II was experimentally induced to have diabetes mellitus using alloxan, a chemical agent that is more or less safe and permanent, and group III was experimentally induced to have diabetes but treated with ascorbic acid (Vit. C), as an antioxidant drug. All rat groups were sacrificed and tissue samples from their parotid and sublingual glands were prepared. The tissue samples studded histologically using Haematoxylin and Eosin (H&E) stain, histopathologically using Periodic acid Schiff's (PAS) reaction to detect carbohydrates, and immunohistochemically using Beta cell lymphoma-2 (BCL2) as an anti-apoptotic marker. Based on the histology, histopathological, and immunohistochemical significant lesions, the obtained results showed common alterations in the parotid and sublingual glands tissues of diabetic rats. However, compared to untreated animals, rats treated with the antioxidant vitamin C showed fewer tissue changes and lesions. According to the study's findings, vitamin C functions as a helpful antioxidant to assist avoid complications from diabetes.

Molecular aspects of predicting recurrence of retroperitoneal extraorgan liposarcomas

Purpose of the study. The aim of the work was to optimize the prognosis of recurrences of retroperitoneal extraorgan liposarcomas of various degrees of differentiation by detecting microRNAs (miRNAs) of tumor cells with differential expression depending on the course of the malignant disease.Materials and methods. The study was conducted on 96 patients: 19 patients with lipomas and 77 patients with retroperitoneal extraorgan liposarcomas. Tissue samples were obtained during the operation. Among patients with malignant diseases, highly differentiated liposarcoma (VDLS) was detected in 51 patients, and dedifferentiated (DDLS) in 26 patients. miRNA expression levels in tumor cells were determined using real-time PCR.Results. During the follow-up period, relapses of malignant disease were detected in 25 (32,5%) people: 13 (25,5%) among patients with VDLS and 12 (46,2%) among patients with DDLS. In DDLS, characterized by a more aggressive course, there was an increased level of expression of miRNA155, -21, -26a2 and a decrease in expression of miRNA15a and -34 compared with patients with VDLS. In patients with recurrent retroperitoneal liposarcomas, the expression of miRNA15a was initially 83% lower in tumor cells of surgical samples (p<0,001), iRNA34 by 79% (p< 0,001), and the expression activity of miRNA155 was 7,54 times higher (p<0,001) and miRNA26a2 by 6,93 times (p<0,001).Conclusion. The determination of the expression of miRNA155, -26a2, -34, -15a in tumor cells during surgical treatment of patients with retroperitoneal liposarcomas is an effective way to assess the risk of recurrence and is relevant for determining the tactics of further treatment of patients.