Goblet Cells Research Articles

Birth cohort effects in appendiceal adenocarcinoma incidence across the United States.

90 Background: Appendiceal adenocarcinoma incidence rates are increasing across all age groups, sexes and histological subtypes in the United States. Birth cohort patterns of appendiceal adenocarcinomas can provide us with new, etiologic clues into these rising rates but have not been examined for this rare malignant tumor type. We estimated appendiceal adenocarcinoma incidence rates across birth cohorts in the United States. Methods: Using the National Cancer Institute’s Surveillance, Epidemiology, and End Results (SEER) Program data from eight population-based cancer registries, we identified a total of 4,858 persons aged 20+ years when diagnosed with a pathologically-confirmed primary appendiceal adenocarcinoma (non-mucinous, mucinous, goblet cell, signet ring cell carcinoma) between 1975 and 2019. Birth cohorts (1900-1904 to 1990-1994) were created using five-year age groups and time periods. The ratio of age-specific incidence rates was estimated in each birth cohort relative to the 1945-1949 birth cohort and reported as incidence rate ratios (IRRs) with 95% confidence intervals (CI). Results: Compared to persons born in 1945-1949, appendiceal adenocarcinoma incidence rates more than tripled among persons born in 1980-1984 (IRR 3.41, 95%CI 2.54-4.56) and quadrupled among persons born in 1985-1989 (IRR 4.62, 95%CI 3.12-6.82). For all tumor histological types, age-specific appendiceal adenocarcinoma incidence rates increased across successive birth cohorts after 1945-1949—although to varying degrees reported across histological types (e.g., 1980-1984: mucinous adenocarcinoma: IRR 2.71, 95% CI 1.73-4.26; non-mucinous adenocarcinoma: IRR 2.60, 95% CI 1.46-4.63; goblet cell adenocarcinoma: IRR 4.95, 95% CI 2.77-8.85). Conclusions: There are strong, yet distinct birth cohort effects for appendiceal adenocarcinomas across histological subtypes that remain unexplained. These patterns signal that recent exposure changes may be influencing this increasing disease risk for Generation X and Millennials now entering mid-adulthood. With the trend toward more non-operative management of appendicitis, another significant consideration for providers is to keep occult appendiceal tumors in the differential diagnosis of patients who present in this way. The current absence of appendiceal cancer prevention and screening modalities emphasizes the importance of efforts to support earlier detection in a rare malignancy where clinical trials have been historically very limited. Moreover, as similar trends have been reported in other gastrointestinal cancers, this is also suggestive of potential shared exposures contributing to this rising cancer burden across generations.

Cumulative experience meets modern science: Remarkable effects of TongXieYaoFang formula on facilitating intestinal mucosal healing and secretory function.

TongXieYaoFang (TXYF), a classical formula used in Traditional Chinese Medicine, is renowned for its efficacy in treating chronic abdominal pain and diarrhoea. Modern research suggests that fundamental relief from these symptoms depends on complete intestinal mucosal healing, which normalises gut secretory functions. Consensus between traditional and modern medical theories indicates that TXYF is particularly suitable for treating the remission phase of ulcerative colitis (UC). Unfortunately, its potential in the remission phase has not received sufficient attention, and its use has been largely limited to a supportive role during the acute phase. This study aimed to elucidate the efficacy of TXYF in promoting intestinal mucosal healing and enhancing gut secretory function during the non-acute damage phase, as well as to identify the underlying mechanisms contributing to its effects. A mouse model of dextran sulphate sodium salt (DSS)-induced colitis was optimised to specifically evaluate the effects of TXYF on mucosal healing during the repair phase. The effects of TXYF on murine colon function were assessed by measuring faecal pellet count and water content, and further evaluated through immunohistochemical analyses. The underlying mechanisms of action of TXYF were elucidated using mouse intestinal organoid cultures, intestinal stem cell (ISCs) transplantation, immunofluorescence, and western blotting. Active components of TXYF were identified via LC-MS/MS analysis and integrated with network pharmacology for bioinformatics assessment. TXYF significantly promoted mucosal healing, as reflected by reduced disease activity scores, increased colon length, enhanced epithelial proliferation, and decreased histological damage. Furthermore, TXYF enhanced the recovery of critical intestinal functions, including barrier integrity, absorption, secretion, and motility. Notably, the improvement in the secretory function was particularly pronounced. Mechanistically, these therapeutic effects were mediated by the upregulation of the Atonal homolog 1/SAM pointed domain containing ETS transcription factor/Mucin 2 pathway, which facilitates the differentiation and maturation of ISCs into goblet cells, thereby contributing to both mucosal repair and enhanced secretory function. Our study demonstrated that TXYF significantly promotes intestinal mucosal healing and enhances secretory function. These findings offer a solid basis for exploring the potential applications of TXYF in UC management during the remission phase.

Physiology, leptin gene expression, and intestinal morphology of pinhead and starved milkfish (Chanos chanos).

"Pinhead" is an abnormal condition of farmed fish which is rarely studied, albeit well known among fish culturists, and is characterized by extreme emaciation and anorexia. In this study, the potential impacts of pinhead condition in milkfish were analyzed and compared to fed, healthy, and a group starved for four weeks. The condition factor and hepatosomatic index of pinhead milkfish were significantly lower compared with fed, healthy individuals. Abnormal plasma osmolality and muscle water content in pinhead milkfish indicated an imbalance in their internal water content. The anorexigenic hormone, leptin A was highly expressed in liver of pinhead milkfish, which could be related to their lack of appetite. Meanwhile, the hepatosomatic index, intestinal somatic index, enterocyte height, number of villi and goblet cells, Na+/K+- ATPase activity, and intestinal protein content of the pinhead milkfish were similar to those of the 4-week starved individuals. Taken together, our results characterized key physiological parameters of pinhead milkfish for the first time. Further investigation is required to understand how environmental or artificial stress can lead to the occurrence of pinhead milkfish, and to develop methods for alleviating this condition.

Bacillus coagulans alleviates intestinal barrier injury induced by Klebsiella pneumoniae in rabbits by regulating the TLR4/MyD88/NF-κB signalling pathway.

Probiotics effectively alleviate host diarrhoea, but the specific mechanism is not clear. Therefore, we explored the protective mechanism of Bacillus coagulans (BC) on intestinal barrier injury induced by Klebsiella pneumoniae (K. pneumoniae) in rabbits by HE, immunofluorescence and 16S rRNA. The results showed that BC pretreatment alleviated the changes in average daily gain, average daily feed intake and FCR caused by K. pneumoniae in rabbits. Moreover, BC alleviated the inflammatory cell infiltration, intestinal villus reduction, crypt deepening and goblet cell reduction caused by K. pneumoniae in rabbits. Further research revealed that BC improved the intestinal barrier by improving the mechanical barrier, chemical barrier, immune barrier and microbial barrier. Specifically, BC improved the intestinal mechanical barrier by improving the intestinal structure, increasing the protein expression of PCNA, increasing the number of goblet cells, and altering the expression of occludin, claudin-1 and ZO-1. BC improved the intestinal chemical barrier by regulating the expression of MUC1 and MUC2 and inhibited the TLR4/MyD88/NF-κB signalling pathway by altering the expression levels of the inflammatory factors IL-1β, IL-6 and TNF-α, thus optimizing the intestinal immune barrier. In addition, adding BC to the diet improved the intestinal microbial barrier of rabbits by reducing the abundance of harmful bacteria and increasing the abundance of beneficial bacteria. In summary, BC protects against K. pneumoniae-induced intestinal barrier damage by improving intestinal morphology, mitigating the inflammatory response and regulating the microbial composition. Among the pretreatments, the pretreatment effect of 1 × 106 CFU/g was the best. This study provides a theoretical basis for the use of BC to prevent and treat diarrhoea caused by K. pneumoniae in rabbits.

Protective effects of carnosic acid on growth performance, intestinal barrier, and cecal microbiota in yellow-feathered broilers under lipopolysaccharide challenge.

This research was performed to investigate protective effects of carnosic acid on growth performance, intestinal barrier, and cecal microbiota of lipopolysaccharide-challenged broilers. Three hundred 1-day-old yellow-feathered broilers (male) were allocated randomly into 5 treatments, with 6 replicates per treatment, and 10 birds per replicate cage. Birds in both the control group (CON) and the lipopolysaccharide-challenged group were provided with a basal diet, while others were fed a basal diet supplemented with 20, 40, and 60 mg/kg carnosic acid (CA20, CA40, CA60), respectively. At 17, 19, and 21 days of age, birds were injected intraperitoneally with lipopolysaccharide (500 μg/kg body weight), except those in CON, which were injected with saline. Compared with challenged birds, the CA20, CA40, and CA60 increased (P < 0.05) the final body weight, average daily gain, and average daily feed intake, and the CA40 and CA60 also decreased diarrhea rate. Compared with challenged birds, carnosic acid reduced (P < 0.05) plasmal levels of D-lactic acid and endotoxin, increased (P < 0.05) the villus height to crypt depth ratio, and the number of goblet cells in duodenum. The CA40 and CA60 elevated (P < 0.05) relative expression of cell junction proteins (Claudin-1/-2 and ZO-1/-2/-3) and MUC-2 in duodenum, while decreased (P < 0.05) relative expression of TLR2, TLR4, and the concentrations of IL-6, IL-10, TNF-α, TGF-β1 in duodenum. CA40 also increased (P < 0.05) the α-diversity of the cecal microbiota and boosted (P < 0.05) the relative abundance of beneficial phyla and genera, particularly Firmicutes, Anaerofilum, and Papilibacter. In conclusion, dietary supplementation with carnosic acid showed protective effects on the growth performance and intestinal health in challenged broilers by down-regulating the expression of TLRs (TLR2/4) and inhibiting the production of inflammatory cytokines, strengthening the tight junction in intestinal epithelial cells, and enhancing the diversity of microbiota and the relative abundance of beneficial bacteria. When supplemented to diet of broilers, 40 mg/kg carnosic acid was recommended.

Anti-inflammatory, immunostimulant and antimicrobial properties of tannic acid in the diet of Oreochromis niloticus infected with Aeromonas hydrophila.

The study aimed to assess the impact of dietary supplementation with tannic acid on the growth, health, and survival of Oreochromis niloticus following exposure to Aeromonas hydrophila. A total of 320 fish were divided into 16 tanks and assigned to four treatment groups: feed with 0.2% tannic acid (TA0.2%), 0.4% tannic acid (TA0.4%), 0.8% tannic acid (TA0.8%), or no tannic acid (Control0%), with each treatment replicated four times, over a 50-day period. At the end of the 50-day period, biological samples were collected from the fish, which were then intraperitoneally injected with A. hydrophila. No significant differences in growth performance were detected between treatments. As expected, levels of total leukocytes, lymphocytes, monocytes, hemoglobin, and mean corpuscular hemoglobin concentration (MCHC) were notably higher in the blood of the fish after infection, regardless of the treatment received. During both the pre- and post-infection periods, monocytes were more abundant in the TA0.2% and TA0.8% treatments compared to the TA0.4% treatment. Additionally, there was a significant interaction between the factors affecting thrombocytes, neutrophils, basophils, hemoglobin, and MCHC. Thrombocytosis and neutrophilia were significantly greater in the TA0.8% treatment pre-infection than in the post-infection and control group. Conversely, a higher number of basophils were observed in the post-infection period in the TA0.8% treatment group compared to the pre-infection period. Total plasma protein levels decreased significantly in the post-infection period, regardless of tannic acid supplementation levels, while immunoglobulin levels increased after exposure to A. hydrophila. Histological analyses revealed a significant increase in the perimeter and number of intestinal villi in the TA0.4% treatment group before infection. The number of goblet cells also increased in the control group (0%), TA0.4%, and TA0.8% before infection. In splenic tissue, the TA0.4% treatment resulted in a reduction in eosinophilic and mononuclear infiltrates, as well as decreased congestion and vacuolation. Hemosiderin levels were lower in the TA0.4% and TA0.2% treatment groups. In the liver, lymphocytic infiltrates were reduced in the TA0.2% and TA0.4% treatment groups, and portal vein congestion was decreased in the TA0.2% post-infection and TA0.4% pre-infection groups. Post-infection survival rates were significantly higher (p<0.05) in the TA0.4% treatment group (91%) compared to the TA0.8% treatment group (85%) and the control group (71%). The results of the present study show that tannic acid has a positive effect on the immune system of Nile tilapia. This is supported by improvements in innate immunity in the blood, as well as the antimicrobial and anti-inflammatory effects seen in histological analyses. Therefore, it is suggested to use a 0.4% tannic acid dose for dietary supplementation of Nile tilapia, along with further studies on the potential benefits of this food additive for tilapia.

Curcumin attenuates ulcerative colitis via regulation of Sphingosine kinases 1/NF-κB signaling pathway.

Curcumin, a compound from Curcuma longa L., has significant anti-inflammatory properties. However, the mechanisms underlying its anti-inflammatory activity in dextran sodium sulfate (DSS)-induced ulcerative colitis (UC) remain inadequately understood. This study aimed to further elucidate the molecular mechanisms of curcumin DSS-induced UC mice. Our data showed that curcumin alleviated DSS-induced colitis by reducing intestinal damage and inflammation, increasing goblet cells in colon tissues. Enzyme-linked immunosorbent assay revealed that curcumin reduced the expression of inflammatory cytokines (tumor necrosis factor-alpha, interleukin-1β, and interleukin-8) in serum and myeloperoxidase in colon tissues. A comprehensive analysis integrating network pharmacology and RNA sequencing (RNA-seq) revealed significant enrichment of the nuclear factor kappa B (NF-κB) signaling pathways. Notably, RNA-seq analysis demonstrated that curcumin significantly downregulated the mRNA expression of sphingosine kinase 1 (SphK1). Furthermore, molecular docking analysis showed that curcumin can bind to SphK1 and NF-κB. Additionally, curcumin was found to inhibit the activation of the SphK1/NF-κB signaling pathway in DSS-induced UC colon tissue. This study addresses pharmacologic and mechanistic perspectives of curcumin that ameliorates DSS-induced UC and inflammatory response.

Impact of fermented bamboo powder on the morphology and physiology of the gastrointestinal tract in yellow-feather broiler chickens.

Bamboo powder, a novel ingredient, is gaining recognition for its potential as a dietary supplement in poultry feed. This study aimed to investigate the effects of fermented bamboo powder (FBP) on antioxidant status, gut hormone activities, intestinal digestive enzyme activities, gut morphological structure, gastrointestinal development, and the expression of nutritional transporter genes in dwarf yellow-feather broiler chickens. A total of 600 healthy 1-day-old chicks were allocated randomly into two groups, with 10 replicates per group and 30 chicks in each replicate. The control group was provided with a standard basal diet, whereas the experimental group received the same basal diet supplemented with 1.0, 2.0, 4.0, and 6.0 g/kg of fermented bamboo powder (FBP) in four phases: Phase I (days 1-22), Phase II (days 23-45), Phase III (days 46-60), and Phase IV (days 61-77). Phases I and II were categorized as the pretreatment period (days 0-45), while Phases III and IV represented the experimental period (days 46-77). Tissue samples were collected during Phase IV for further analysis. After 77 days of feeding, results revealed that FBP supplementation significantly enhanced the levels of gastrointestinal hormones (Glucagon-like peptide 1, Peptide YY, Cholecystokinin, and 5-hydroxytryptamine) in the duodenum, jejunum, and ileum. Similarly, the activities of digestive enzymes (protease, chymotrypsin, trypsin, and amylase) were significantly increased in the small intestine. It also improved gut morphology by increasing villus height, crypt depth, and goblet cell counts in the duodenum, jejunum, and ileum. Additionally, antioxidant enzyme activities (superoxide dismutase, glutathione peroxidase, and catalase) were significantly increased, while malondialdehyde content was significantly decreased in the jejunum. Additionally, FBP supplementation significantly enhanced gizzard development. Overall, FBP supplementation modulated gut hormones and enzymes, enhanced gut morphology and promoted antioxidant status and gene expression related to nutrient transport and antioxidant defenses in broiler chickens. These findings suggest that FBP has the potential as a beneficial dietary supplement in poultry feed.

Management trends and outcomes of appendiceal cancers: A National Cancer Database study.

810 Background: Appendiceal tumors encompass a variety of histological types that exhibit different behaviors. The current study aimed to investigate and compare overall survival, predictive factors for overall survival, and management of histological variations of appendiceal tumors. Methods: The National Cancer Data Bank (NCDB) database (2004–2020) was evaluated to include patients with appendiceal tumors histologies including goblet cell adenocarcinoma (GCA), neuroendocrine neoplasm (NEN), non-mucinous adenocarcinoma (NMA), and mucinous adenocarcinoma (MA). Univariate and multivariable analyses were conducted. Results: The GCA, MA, NEN, and NMA groups consist of 6,111, 16,471, 19,199, and 11,065 patients, respectively. The NMA and NEN groups had significantly lowest and higher overall survival compared to other types (p&lt;0.001 for both) (GCC: 143.27 (140.23-146.30), MA: 111.91 (109.996-113.832), NEN: 170.88 (168.56-173.20), and NMA: 101.399 (99.132-103.666) months). The independent predicting factors of worse overall survival were age (HR: 1.03 (1.03-1.04)), black race (HR: 1.24 (1.08-1.43), ref: white race), fourth median income (HR: 0.84 (0.74-0.96), ref: first median), Charleson index comorbidity score (HR: 1.3 (1.22-1.38)), lymphovascular invasion (HR: 1.28 (1.15-1.42)), pathologic stage 2 (HR: 1.32 (1.12-1.57)), stage 3 (HR: 1.99 (1.66-2.40)), stage 4 (HR: 4.20 (3.47-5.07)), ref: stage 1, intraoperative systemic therapy (HR: 0.52 (0.39-0.70)), positive surgical margin (1.55 (1.39-1.73)), number of positive lymph nodes (HR:1.06 (1.05-1.07)), moderately differentiated tumors (HR:1.43 (1.25-1.63)), poorly differentiated (HR: 1.94 (1.69-2.24)), undifferentiated (HR: 1.48 (1.14-1.91)), ref: well differentiated, and NMA type (HR:1.30 (1.10-1.55)). Furthermore, in terms of type of surgery, significantly higher patients of GCA (54.6%) and NMA (55.2%) groups underwent subtotal colectomy compared to other groups. Regarding systemic therapy, significantly higher patients of MA group received intraoperative, and both neoadjuvant and adjuvant therapy compared to other groups. Also, NEN group had significantly higher patients who underwent surgery with laparoscopic approach compared to others. Conclusions: This study reveals that different types of appendiceal tumors have distinct characteristics in regard to overall survival outcomes and treatment approaches. NENs showed the highest survival rates, while NMAs had the lowest. These results underscore the need for personalized treatment strategies for each tumor type to improve patient outcomes.

Histopathological and histochemical analysis of the digestive tract of adult horseshoe crabs under mercury stress.

Mercury (Hg) contamination is a major environmental concern that continuously impacts marine organisms, including the mangrove horseshoe crab (Carcinoscorpius rotundicauda). As a vulnerable species, C. rotundicauda holds significant ecological and evolutionary value, making it particularly susceptible to Hg pollution and deserving of focused conservation efforts. This study presents the first comprehensive histopathological and histochemical analysis of mercury toxicity in the digestive tract of adult C. rotundicauda. Through both qualitative and quantitative analyses of histopathological and histochemical alterations, we evaluated the effects of acute Hg exposure (0.5 mg/L and 5 mg/L) on the esophagus, stomach, and intestines over time. The results revealed significant dose- and time-dependent tissue damage, with the stomach exhibiting the most pronounced alterations, including epithelial detachment, vacuolation caused by fibers degradation in the loose connective tissue, and muscle layer disruption, followed by the esophagus and intestines. Additionally, mucous cell density in the hindgut submucosa decreased by 30 % after 96 h of acute Hg exposure (0.5 and 5 mg/L HgCl2), indicating a weakened protective barrier. The tissue damage of the digestive tract may further have a negative impact on the health of the adult horseshoe crab, thus threatening the survival of C. rotundicauda population and reducing the biodiversity of the mangrove ecosystem. These findings underscore the critical threat posed by Hg pollution to the digestive system of C. rotundicauda and offer novel insights into the toxicological impact on marine invertebrates. Considering the tissue-damaging effects of Hg on adult horseshoe crabs, this study underscores the importance of regular monitoring of Hg contamination in natural habitats. The results offer valuable guidance for future habitat restoration and effective management of existing habitats.

Anti-IL-5 treatment, but not neutrophil interference, attenuates inflammation in a mixed granulocytic asthma mouse model, elicited by air pollution

IntroductionDiesel exhaust particles (DEP) have been proven to aggravate asthma pathogenesis. We previously demonstrated that concurrent exposure to house dust mite (HDM) and DEP in mice increases both eosinophils and neutrophils in bronchoalveolar lavage fluid (BALF) and also results in higher levels of neutrophil-recruiting chemokines and neutrophil extracellular trap (NET) formation compared to sole HDM, sole DEP or saline exposure. We aimed to evaluate whether treatment with anti-IL-5 can alleviate the asthmatic features in this mixed granulocytic asthma model. Moreover, we aimed to unravel whether neutrophils modulate the DEP-aggravated eosinophilic airway inflammation.Material and methodsFemale C57BL6/J mice were intranasally exposed to saline or HDM and DEP for 3 weeks (subacute model). Interference with eosinophils was performed by intraperitoneal administration of anti-IL-5 (TRFK5), which neutralizes IL-5. Interference with neutrophils and neutrophil elastase was performed by intraperitoneal anti-Ly6G and sivelestat administration, respectively. Outcome parameters included eosinophils subsets (homeostatic EOS and inflammatory EOS), proinflammatory cytokines, goblet cell hyperplasia and airway hyperresponsiveness.ResultsThe administration of anti-IL-5 significantly decreased eosinophilic responses, affecting both inflammatory and homeostatic eosinophil subsets, upon subacute HDM + DEP exposure while BAL neutrophils, NET formation and other asthma features remained present. Neutrophils were significantly reduced after anti-Ly6G administration in BALF, lung and blood without affecting the eosinophilic inflammation upon HDM + DEP exposure. Sivelestat treatment tended to decrease BALF inflammation, including eosinophils, upon HDM + DEP exposure, but did not affect lung inflammation.ConclusionInhibition of IL-5 signalling, but not neutrophil interventions, significantly attenuates eosinophilic inflammation in a mouse model of mixed granulocytic asthma, elicited by air pollution exposure.

Assessment of a semiquantitative scoring system for mild-to-moderate gill lesions in Atlantic salmon reared in recirculating aquaculture systems in Norway.

Compromised gill health is a critical cause of forfeited welfare in Atlantic salmon farming. Detecting and quantifying the early onset of gill disease is important to reveal initial inflicting stimuli. We collected gill samples of 45 Atlantic salmon from 2 commercial recirculating aquaculture systems (RASs) spanning fry-to-market-size fish with no clinical signs of gill disease. Gill samples were assessed histologically by 3 independent raters with different levels of experience. Semiquantitative scoring for 7 types of gill changes was carried out for 10 filaments per gill (450 filaments total) over 3 rounds on anonymized samples. Scores were summarized for each type of gill change. The assumed clinical relevance for each change was transformed into a category score, followed by an assessment of agreement within (intra) and between (inter) raters. A generalized linear model estimated the difference in score levels between raters. For each rater, intra-rater agreement was high for 6 gill changes and moderate for 1 gill change. Inter-rater agreement was moderate to almost-perfect, except for 2 gill changes; generalized linear model regression revealed systematic differences in score usage between the raters. Our scoring protocol worked satisfactorily for mucous cell amount, lamellar clubbing, lamellar hypertrophy and/or hyperplasia, and aneurysms, despite different levels of expertise in histologic evaluation. Intra-rater agreement was consistent, but differences existed for interlamellar hypercellularity, lamellar inflammation, and degeneration. Scoring subclinical gill changes is a challenge, and our scoring system for mild-to-moderate lesions may enable early intervention to limit the detrimental effects of poor gill health in RAS farming.

Airway Basal Stem Cells Inflammatory Alterations in COVID-19 and Mitigation by Mesenchymal Stem Cells.

SARS-CoV-2 infection and the resultant COVID-19 pneumonia cause significant damage to the airway and lung epithelium. This damage manifests as mucus hypersecretion, pulmonary inflammation and fibrosis, which often lead to long-term complications collectively referred to as long COVID or post-acute sequelae of COVID-19 (PASC). The airway epithelium, as the first line of defence against respiratory pathogens, depends on airway basal stem cells (BSCs) for regeneration. Alterations in BSCs are associated with impaired epithelial repair and may contribute to the respiratory complications observed in PASC. Given the critical role of BSCs in maintaining epithelial integrity, understanding their alterations in COVID-19 is essential for developing effective therapeutic strategies. This study investigates the intrinsic properties of BSCs derived from COVID-19 patients and evaluates the modulatory effects of mesenchymal stem cells (MSCs). Through a combination of functional assessments and transcriptomic profiling, we identified key phenotypic and molecular deviations in COVID-19 patient-derived BSCs, including goblet cell hyperplasia, inflammation and fibrosis, which may underlie their contribution to PASC. Notably, MSC co-culture significantly mitigated these adverse effects, potentially through modulation of the interferon signalling pathway. This is the first study to isolate BSCs from COVID-19 patients in the Chinese population and establish a COVID-19 BSC-based xenograft model. Our findings reveal critical insights into the role of BSCs in epithelial repair and their inflammatory alterations in COVID-19 pathology, with potential relevance to PASC and virus-induced respiratory sequelae. Additionally, our study highlights MSC-based therapies as a promising strategy to address respiratory sequelae and persistent symptoms.

Migrasomes derived from human umbilical cord mesenchymal stem cells: a new therapeutic agent for ovalbumin-induced asthma in mice

BackgroundAsthma is a prevalent respiratory disease, and its management remains largely unsatisfactory. Mesenchymal stem cells (MSCs) have been demonstrated to be efficacious in reducing airway inflammation in experimental allergic diseases, representing a potential alternative treatment for asthma. Migrasomes are recently identified extracellular vesicles (EVs) generated in migrating cells and facilitate intercellular communication. The objective of this study was to investigate the therapeutic effects of migrasomes obtained from MSC in a model of asthma.MethodsMigrasomes produced by human umbilical cord MSCs (hUCMSCs) were isolated by sequential centrifugation. Characterization of hUCMSC-derived migrasomes were carried out by transmission electron microscopy and western blot analysis. The therapeutic effects of migrasomes on airway inflammation in ovalbumin (OVA)-induced asthmatic mice were evaluated by hematoxylin-eosin (HE) and periodic-acid schiff (PAS) staining, and their mechanism were further testified by immunofluorescent staining, real-time PCR and flow cytometry.ResultsHere, we showed that inhibition of migrasomes’ production dramatically impaired the anti-inflammatory effects of hUCMSCs in OVA animals, as evidenced by a notable increase in both the infiltration of inflammatory cells and the number of epithelial goblet cells. We successfully isolated hUCMSC-migrasomes, which were morphologically intact and positive for the specific migrasomes markers. The administration of hUCMSC-migrasomes was observed to significantly ameliorate the symptoms of airway inflammation and mucus production in asthmatic mice. Additionally, the expression of Th2 cytokines (IL-4, IL-5 and IL-13) were found to be reduced, while the activation of dendritic cells (DCs) was inhibited. HUCMSC-migrasomes could possibly be delivered to lung region after injection, and were able to be taken in by DCs both in vivo and in vitro. Notably, in vitro, migraosmes decreased the capacity of BMDCs to stimulate OVA-specific Th2-cell responses. More importantly, we found that adoptive transfer of hUCMSC-migrasomes-treated BMDCs was sufficient to protect mice from allergic airway inflammation. In addition, we found that hUCMSC-migrasomes inhibited the receptor for advanced glycation end-products (RAGE) signal in OVA-treated BMDCs in vitro and in asthma mice lung in vivo.ConclusionOur results provided the first evidence that hUCMSC-migrasomes possess anti-inflammatory properties in OVA-induced allergic mice, which may provide a novel “MSC-cell free” therapeutic agent for the management of asthma.

Yeast cell wall polysaccharides accelerate yet in-feed antibiotic delays intestinal development and maturation via modulating gut microbiome in chickens

BackgroundIt is important to promote intestinal development and maturation of chicks for feed digestion and utilization, intestinal health, and disease resistance. This study aimed to investigate the effects of dietary yeast cell wall polysaccharides (YCWP) addition on intestinal development and maturation of chickens and its potential action mechanism.Methods180 one-day-old male Arbor Acres broilers were randomly assigned to three groups containing control (basal diets without any antibiotics or anticoccidial drug), bacitracin methylene disalicylate (BMD)-treated group (50 mg/kg) and YCWP-supplemented group (100 mg/kg).ResultsCompared with control group, in-feed antibiotic BMD continuous administration significantly decreased crypt depth (d 21) and villus height (d 42) along with mucosal maltase activity (d 42) in the ileum (P < 0.05). Also, BMD markedly downregulated gene expression levels of β-catenin, lysozyme, occludin and FABP-2 (d 21) and innate immune related genes CD83 and MHC-I mRNA levels (d 42, P < 0.05), and decreased goblet cell counts in the ileum of chickens (d 21 and d 42, P < 0.05). While, TLR-2, TLR-6 and iNOS mRNA abundances were notably upregulated by BMD treatment (d 42, P < 0.05). Nevertheless, dietary YCWP addition significantly increased the ratio of villus height to crypt depth (d 21), villus surface area (d 21 and d 42), ileal alkaline phosphatase and maltase activities as well as goblet cell (d 21 and d 42) and IgA-producing plasma cell numbers as compared to BMD treatment (d 21, P < 0.05). YCWP addition also upregulated gene expression levels of Lgr5, Wnt/β-catenin signaling pathway related gene (Wnt3, β-catenin, d 21; β-catenin, d 42), intestinal cells proliferation marker Ki-67 and barrier function related genes (occludin, d 21 and d 42, P < 0.05). Moreover, YCWP significantly increased antigen presenting cell marker related genes (MHC-II, d 21; CD83 and MHC-I, d 42), TLR-1, TLR-2 and TLR-6 mRNA levels (d 21, P < 0.05). Cecal microbiome analysis showed that YCWP addition obviously improved cecal microbial composition, as indicated by increasing relative abundance of Fournierella, Psychrobacter and Ruminiclostridium on d 21, and Alistipes and Lactobacillus on d 42, which were positively related with gut development and maturation related indexes (P < 0.05).ConclusionCollectively, YCWP promoted yet antibiotic BMD delayed intestinal morphological and immunological development linked with modulating gut microbiome in chickens.

Promising protective treatment potential of endophytic bacterium Rhizobium aegyptiacum for ulcerative colitis in rats

Abstract Ulcerative colitis (UC) is an inflammatory condition of the intestine, resulting from an increase in oxidative stress and pro-inflammatory mediators. In this study, the extract of endophytic bacterium Rhizobium aegyptiacum was prepared for the first time using liquid chromatography-mass spectrometry (LC-MS). In addition, also for the first time, the protective potential of R. aegyptiacum was revealed using an in vivo rat model of UC. The animals were grouped into four categories: normal control (group I), R. aegyptiacum (group II), acetic acid (AA)-induced UC (group III), and R. aegyptiacum-treated AA-induced UC (group IV). In group IV, R. aegyptiacum was administered at 0.2 mg/kg daily for one week before and two weeks after the induction of UC. After sacrificing the rats on the last day of the experiment, colon tissues were collected and subjected to histological, immunohistochemical, and biochemical investigations. There was a remarkable improvement in the histological findings of the colon tissues in group IV, as revealed by hematoxylin and eosin (H&amp;E) staining, Masson’s trichrome staining, and periodic acid-Schiff (PAS) staining. Normal mucosal surfaces covered with a straight, intact, and thin brush border were revealed. Goblet cells appeared magenta in color, and there was a significant decrease in the distribution of collagen fibers in the mucosa and submucosal connective tissues. All these findings were comparable to the respective characteristics of the control group. Regarding cyclooxygenase-2 (COX-2) immunostaining, a weak immune reaction was shown in most cells. Moreover, the colon tissues were examined using a scanning electron microscope, which confirmed the results of histological assessment. A regular polygonal unit pattern was seen with crypt orifices of different sizes and numerous goblet cells. Furthermore, the levels of catalase (CAT), myeloperoxidase (MPO), nitric oxide (NO), interleukin-6 (IL-6), and interlukin-1β (IL-1β) were determined in the colonic tissues of the different groups using colorimetric assay and enzyme-linked immunosorbent assay (ELISA). In comparison with group III, group IV exhibited a significant rise (P&lt;0.05) in the CAT level but a substantial decline (P&lt;0.05) in the NO, MPO, and inflammatory cytokine (IL-6 and IL-1 β) levels. Based on reverse transcription-quantitative polymerase chain reaction (RT-qPCR), the tumor necrosis factor-α (TNF-α) gene expression was upregulated in group III, which was significantly downregulated (P&lt;0.05) by treatment with R. aegyptiacum in group IV. On the contrary, the heme oxygenase-1 (HO-1) gene was substantially upregulated in group IV. Our findings imply that the oral consumption of R. aegyptiacum ameliorates AA-induced UC in rats by restoring and reestablishing the mucosal integrity, in addition to its anti-oxidant and anti-inflammatory effects. Accordingly, R. aegyptiacum is potentially effective and beneficial in human UC therapy, which needs to be further investigated in future work.

Metabolic and Intestinal Morphometric Responses of Nile Tilapia Fed Diets Containing Soybean and Protease

Proteases facilitate the breakdown of components associated with antinutritional factors. This study evaluated the effects of protease supplementation in Nile tilapia diets on metabolic and intestinal responses. Three diets were tested: soybean meal SM1 (fish meal FM:SM = 1:1), SM2 (1:2), and SM3 (1:3), based on the protein content. All the diets were without or with protease (0.44 g/kg). The fish were randomly allocated to 18 tanks/49 days. In SM3 without protease, an increase in the villus height and width, as well as the number of goblet cells, was observed. Protease supplementation increased the villus height compared to diets without the enzyme. The SM1 with protease resulted in a reduction in the digestive somatic index, but significantly increased the hepatosomatic and intestinal quotient indices. In SM2 with protease, reductions in the amino acid (AA) content and aspartate aminotransferase activity in the liver were observed. In the muscles, supplementation increased the AA concentrations. Higher plasma AA and albumin concentrations were detected in the SM2 and SM3 groups with protease. A significant interaction was found, with SM3 without protease showing lower plasma protein and AA concentrations. In SM2 with protease, higher levels of albumin and globulin were recorded. Protease in soybean meal diets improved the intestinal health and metabolism in tilapia.

Label-free quantitative imaging of conjunctival goblet cells.

To introduce and validate quantitative oblique back-illumination microscopy (qOBM) as a label-free, high-contrast imaging technique for visualizing conjunctival goblet cells (GCs) and assessing their functional changes. qOBM was developed in conjunction with moxifloxacin-based fluorescence microscopy (MBFM), which was used for validating GC imaging. Initial validation was conducted with polystyrene beads, followed by testing on normal mouse conjunctiva under both ex-vivo and in-vivo conditions. Longitudinal qOBM imaging was performed on ex-vivo mouse conjunctiva exposed to hyperosmotic stress (induced by 1000 mOsm/kg NaCl solution) and normal saline (300 mOsm/kg balanced salt solution, BSS). Imaging was conducted at baseline and at 15- and 30-minute instillation. Results were compared to those of MBFM and periodic acid-Schiff (PAS) staining. A similar longitudinal study was performed in-vivo, and the outcomes were analyzed. qOBM accurately imaged polystyrene beads, with measured phase delays matching theoretical predictions. In normal mouse conjunctiva, qOBM visualized GCs in high contrast, confirmed by MBFM, and the average phase delay was 0.59 ± 0.25 radians. Under hyperosmotic stress, qOBM detected a significant reduction in GC phase delays, decreasing to levels of the surrounding tissue (-0.07 ± 0.14 radians). In normal conditions, no notable changes were observed in GCs. In-vivo imaging results were consistent with ex-vivo findings. Statistical analysis further characterized these changes. The results were consistent with MBFM and PAS staining. qOBM is a high-contrast, label-free imaging modality that enables the functional assessment of GCs. This technique holds significant potential for advancing research and clinical diagnostics related to ocular surface diseases.

Histopathology of the tongue in a hamster model of COVID-19

ObjectiveWith altered sense of taste being a common symptom of coronavirus disease 2019 (COVID-19), the main objective was to investigate the presence and distribution of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) within the tongue over the course of infection.MethodsGolden Syrian hamsters were inoculated intranasally with SARS-CoV-2 and tongues were collected at 2, 3, 5, 8, 17, 21, 35, and 42 days post-infection (dpi) for analysis. In order to test for gross changes in the tongue, the papillae of the tongue were counted. Paraffin-embedded thin sections of the tongues were labeled for the presence of SARS-CoV-2 antigen.ResultsThere was no difference in fungiform or filiform papillae density throughout the course of infection. SARS-CoV-2 antigen was observed in the vallate papillae taste buds (3–35 dpi) and autonomic ganglia (5–35 dpi), as well as in the serous and mucous salivary glands of the posterior tongue (2–42 dpi).ConclusionThe presence and distribution of SARS-CoV-2 suggest that the virus could cause taste disturbance by infecting the vallate papillae taste buds. This effect could be exacerbated by a diminished secretion of saliva caused by infection of the serous salivary glands and the autonomic ganglia which innervate them.

Diffuse Cyclin D1 and SPINK1 Expression in Gastric Oxyntic Gland Neoplasms: Promising Diagnostic Markers Identified by Spatial Transcriptome Analysis.

Oxyntic gland neoplasms typically arise in Helicobacter pylori-naïve stomachs and are composed predominantly of chief cells, with a smaller component of parietal cells. The pathologic diagnosis can be challenging due to minimal cellular atypia. Especially in biopsy specimens with limited tumor volume or when pathologists have limited experience in diagnosing this neoplasm, distinguishing it from normal oxyntic glands can be difficult, and no reliable diagnostic markers are currently available. In this study, single-cell spatial transcriptome analysis successfully identified significant upregulation of CCND1 and SPINK1 in all six analyzed cases of oxyntic gland neoplasms compared to normal oxyntic glands. Immunohistochemical analysis confirmed this finding in 21 endoscopically resected cases of oxyntic gland neoplasms, demonstrating that cyclin D1 and SPINK1 were diffusely expressed in oxyntic gland neoplasms, whereas their expression was scarcely observed in normal oxyntic glands with a few of them showing weak to moderate staining. Even in biopsy specimens, these two markers highlighted the tumor areas and clearly distinguished neoplastic from normal oxyntic glands. Nonneoplastic foveolar epithelia and mucous neck cells also showed positive staining for both cyclin D1 and SPINK1. Additionally, mild increase in cyclin D1 expression and patchy or mosaic expression of SPINK1 was observed in fundic gland polyps, Helicobacter pylori -associated gastritis and pyloric gland adenomas, while diffuse staining pattern was specific to oxyntic gland neoplasms. These observations suggest that cyclin D1 and SPINK1 are reliable markers in differentiating oxyntic gland neoplasms from nonneoplastic oxyntic glands and pyloric gland adenomas. Cyclin D1 is commonly used for immunostaining in many pathology departments, and due to its higher sensitivity and specificity compared to SPINK1, it is considered the best diagnostic marker.