Stomach Tissue Samples Research Articles

Gastric lesions associated with the infection of Anisakidae nematodes in a dwarf sperm whale Kogia sima (Owen, 1866) stranded in the north coast of Brazil

The present study aimed to describe gastric lesions associated with parasitism by different nematodes of the family Anisakidae in a stranded specimen of dwarf sperm whale (Kogia sima) on the northeast coast of the State of Pará, northern Brazil. Specimens of helminths and samples of stomach tissue were collected from a dwarf sperm whale, stranded on Humaitá beach, State of Pará, Brazil. Stomach showed areas of erosion and ulcers, with the mucosa covered by fibrinonecrotic material containing bacteria and inflammatory infiltrate predominantly comprising polymorphonuclear cells. Granulomas were also found in the submucosa, characterized by central areas of necrosis and hemorrhage, and cross sections of nematodes were observed. Fourth-stage larvae of the genus Pseudoterranova, two morphotypes of fourth-stage larvae of the genus Anisakis and adult specimens of Skrjabinisakis paggiae were morphologically identified. Molecular and phylogenetic analyzes confirmed the identity of the partial sequences of the cox2 mtDNA gene for adult specimens of S. paggiae. This study contributes to our understanding of the distribution of different of anisakids in K. sima and about the gastric lesions associated with these nematodes, in addition to expanding the knowledge about the occurrence of this aquatic mammal recorded for the first time in the northern region of Brazil.

Pathogenesis of Experimental Infection of Nile Tilapia (Oreochromis niloticus) with Nucleospora Braziliensis Pathology and Proteomic of Microsporidia.

The recent discovery of disease caused by Nucleospora braziliensis in Nile tilapia (Oreochromis niloticus) is important as it has highlighted the high prevalence of infection and associated mortality in cultured fish. Thus, this study conducted an experimental infection of this microsporidium to evaluate pathological alterations and conduct proteomic analysis. For pathological observation, samples of brain, eyes, gall bladder, gut, heart, kidney, liver, muscle, skin, spleen, and stomach tissue, were collected, and liquid chromatography-mass spectrometry (LC-MS/MS) was performed for proteomic analysis. The most prevalent lesions were brownish color of the liver, gill filament fusion, gut ischemia, hemorrhage of the lips and fins, hepatomegaly, spleen atrophy, splenomegaly, and stomach congestion. The most common microscopic lesions were degeneration, hemorrhage, and inflammation in the brain, gills, gut, kidney, liver, muscle, spleen, and stomach. The digested peptides were identified by LC-MS/MS and the intersection of each group showed that in the spleen there were 121 exclusive proteins in the infected sample and 252 in the control, while in the kidney, 129 proteins were identified in the infected specimen compared to 83 in the control. In conclusion, this study demonstrates the proteome profile of O. niloticus kidney and spleen tissue in response to infection with N. braziliensis.

Abstract 5583: Helicobacter pylori induces the wee1, promoting immune suppressive in human gastric adenocarcinoma

Abstract Background: Gastric cancer (GC) is the fourth most common cause of cancer-related death worldwide. GC has a dismal 5-year survival rate of about 32%. Helicobacter pylori (H. pylori) infection is the strongest risk factor for gastric carcinogenesis. 75-90% of gastric cancer patients exhibit positive H. pylori serology. WEE1 is a nuclear kinase that regulates cell cycle progression by inhibiting cyclin-dependent kinases. Methods & Results: Analysis of non-cancer normal stomach tissue (n=360) and human GC samples (n=1221) identified that WEE1 mRNA was significantly overexpressed in GC. Using immunohistochemistry (IHC) staining in 41 normal and 44 GC human samples, the data indicated that WEE1 protein was significantly overexpressed in GC. Western blot and immunofluorescence (IF) staining confirmed that H. pylori infection induces WEE1 overexpression in GC cells with an unexpected cytosolic localization of WEE1 in GC cells. Tff1 knockout GC mouse model (Tff1-/-) was infected with mouse-adapted H. pylori strain PMSS1. Western blot and immunohistochemistry data demonstrated that H. pylori infection induced WEE1 expression in Tff1 -/- HGD/cancer mouse gastric tissues compared to control Tff1 -/- mouse stomach. To study how H. pylori infection induces WEE1 in GC, analysis from miRTAR, Target Scan, and miRDB demonstrated that WEE1 is predicted to be targeted by miR-497-5p. Using qRT-PCR analysis, we assessed 30 normal and 30 GC human tissue samples, revealing a pronounced downregulation of miR-497-5p in GC specimens. Western blot analysis confirmed that the restoration of miR-497-5p in GC cells markedly reduced WEE1 protein expression. Interestingly, H. pylori infection further decreased miR-497-5p levels in GC cells, upregulating WEE1 protein expression. Since the miR-497-5P host gene promoter region contains CpG islands, we confirmed that treatment with 5 Azacytidine (demethylating agent) or DNMT1 knockdown induced expression of miR-497-5p with decreased WEE1 level in GC cells. Furthermore, our preliminary results indicate that aberrant WEE1 overexpression in GC demonstrates a significant positive correlation with immune-suppression gene signatures, underscoring its predictive value for unfavorable patient survival outcomes in GC. Using the TFF1 knockout mouse GC model, we found that WEE1 inhibition notably decreases the number and activation of CD4+ and CD8+ T cells in neoplastic gastric tissues. These findings also indicate a crucial relationship between WEE1 and the immunosuppressive microenvironment in GC, establishing WEE1 as a promising therapeutic target to enhance therapeutic efficacy and overcome immunotherapy resistance in GC. Conclusions: Our findings demonstrated a novel mechanism in which H. pylori activates the WEE1 in GC, promoting an immune suppressive microenvironment. WEE1 inhibition with immunotherapy could be a promising therapeutic method for GC patients. Citation Format: Md Obaidul Islam, Krishnapriya Thangaretnam, Heng Lu, Dunfa Peng, Nadeem Sidiq Bhat, Mohammed Soutto, Wael El-Rifai, Zheng Chen. Helicobacter pylori induces the wee1, promoting immune suppressive in human gastric adenocarcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 5583.

INVESTIGATING THE THERAPEUTIC POTENTIAL OF BENIDIPINE IN INDO-METHACIN-INDUCED PEPTIC ULCERS: AN EXPERIMENTAL STUDY IN RATS

Objective: Peptic ulcer disease is a prevalent gastrointestinal disorder associated with escalating global mortality rates and a heightened risk of gastric cancer. This study investigates the therapeutic potential of Benidipine, an L-type calcium channel blocker known for its antioxidant properties, in an indomethacin-induced peptic ulcer model in rats. Methods: Thirty-six male albino Wistar rats were divided into six groups and subjected to various drug treatments, including Indomethacin, Famotidine, and Benidipine. Stomach tissue samples were collected for analysis. Results: Macroscopic examination revealed that indomethacin significantly increased ulcer areas in the stomach tissue, while Benidipine and Famotidine treatments effectively reduced these ulcer areas. Biochemical analysis indicated that Indomethacin-induced oxidative stress, as evidenced by elevated levels of malondialdehyde, a marker of lipid peroxidation, and decreased levels of total glutathione and superoxide dismutase, are crucial antioxidants. However, Benidipine treatment mitigated these changes, restoring antioxidant levels and reducing malondialdehyde levels. Conclusion: This study demonstrates that indomethacin induces gastric damage and oxidative stress, leading to ulcer formation, while Benidipine, with its antioxidant properties, effectively counteracts these effects and exhibits comparable efficacy to Famotidine in ulcer reduction. These findings suggest that Benidipine may hold promise as a therapeutic agent for mitigating peptic ulcers induced by nonsteroidal anti-inflammatory drugs like Indomethacin, warranting further exploration in clinical settings.

GC-MS/MS analysis of chlorpyrifos in forensic samples with varied survival time.

Organophosphorus pesticides are extensively used in the agricultural sector to kill insects, worms, and other pests. Many people may be poisoned by chlorpyrifos either accidentally or intentionally, including accidental, suicidal, and homicidal poisoning cases in India. The effect of chlorpyrifos on human health depends on factors such as the time, amount and frequency of exposure, the individual's health, and certain environmental conditions. The main objective of this investigation is to identify the post-mortem biological sample that shows the longest detection window, enabling precise chlorpyrifos detection in cases of acute poisoning with varying survival durations. Our research focuses on the detection and distribution of chlorpyrifos in cases of acute poisoning using a simple liquid-liquid extraction and GC-MS/MS analysis. We validated the method, which proved to be effective and reliable. Upon examining various organs, we detected the presence of chlorpyrifos in the stomach tissue, liver tissue, kidney tissue, and blood samples of individuals who consumed chlorpyrifos and passed away immediately, as well as in those who survived for the first 3days following ingestion. Analysing urine, blood, and liver tissue from individuals who survived for 3days provided more precise results compared to stomach tissue. Additionally, urine samples played a crucial role in detecting chlorpyrifos in individuals who survived for 4 and 5days. A blood sample is the most suitable post-mortem biological sample for detecting chlorpyrifos in individuals who survived for a duration of 2 to 4days. This finding highlights the significance of analysing urine as a valuable sample type, particularly in determining the presence of chlorpyrifos in cases where individuals have survived for a long period of time before their demise. The experimental data and information provided in this study will serve as a valuable resource for forensic toxicologists.

Ameliorative effect of Silk Fibroin against 5-Fluorouracil (5-FU)-induced gastrointestinal damage in rats

Objective: 5-Fluorouracil (5-FU), a widely administered anti-cancer drug, causes gastrointestinal damage through various mechanisms. This study aimed to investigate the ameliorative effect of silk fibroin (SF), which has anti-inflammatory and antioxidant properties, against 5 FU-induced gastrointestinal damage.
 Materials and methods: Wistar albino rats were divided into 3 groups; Control group, 5- FU group (30mg/kg/day on days 0, 2, 4), and 5-FU+SF (30 mg/kg/day 5-FU on days 0, 2, 4 + 600 mg/kg/day SF for 14 days). At the end of the experiment, stomach and intestinal tissue samples were collected and immunohistochemically iNOS, caspase-3, CD4+ and CD8+ were measured. Besides, structural damage was assessed with hematoxylin-eosin and caspase-3 and 9 activities were evaluated using western blot analysis.
 Results and Discussion: It was determined that iNOS, caspase-3, caspase-9 activities, CD4+ and CD8+ levels and structural damage significantly increased in both stomach and intestinal tissues. Furthermore, elevated markers were regressed almost to control levels in SF administrated group.
 Conclusion: SF may have an ameliorative effect, thus reducing the gastrointestinal damage caused by 5 FU. More clinical and scientific research should be conducted as the silk fibroin may be a suitable candidate against the adverse effects of chemotherapeuic agents.
 Bangladesh Journal of Medical Science Vol. 22 No. 04 October’23 Page : 907-915

P53 amyloid pathology is correlated with higher cancer grade irrespective of the mutant or wild-type form.

p53 (also known as TP53) mutation and amyloid formation are long associated with cancer pathogenesis; however, the direct demonstration of the link between p53 amyloid load and cancer progression is lacking. Using multi-disciplinary techniques and 59 tissues (53 oral and stomach cancer tumor tissue samples from Indian individuals with cancer and six non-cancer oral and stomach tissue samples), we showed that p53 amyloid load and cancer grades are highly correlated. Furthermore, next-generation sequencing (NGS) data suggest that not only mutant p53 (e.g. single-nucleotide variants, deletions, and insertions) but wild-type p53 also formed amyloids either in the nucleus (50%) and/or in the cytoplasm in most cancer tissues. Interestingly, in all these cancer tissues, p53 displays a loss of DNA-binding and transcriptional activities, suggesting that the level of amyloid load correlates with the degree of loss and an increase in cancer grades. The p53 amyloids also sequester higher amounts of the related p63 and p73 (also known as TP63 and TP73, respectively) protein in higher-grade tumor tissues. The data suggest p53 misfolding and/or aggregation, and subsequent amyloid formation, lead to loss of the tumor-suppressive function and the gain of oncogenic function, aggravation of which might determine the cancer grade.

Differential expression of KCNQ1 and ATP4A genes according to the sex and age in the stomach

We compared the expression of six genes in stomach tissue samples between healthy men and women in different age groups to study sexually dimorphic gene expression. Real-Time RT-PCR was used to compare gene expression between men and women. Our results showed that the expression of KCNQ1 (p = 0.01) was significantly higher in non-menopausal women compared to post-menopausal women. In addition, the expression level of the ATP4A gene in men under 35 years was significantly higher than in men above 50 (p = 0.026). Sexually and age dimorphic gene expression in some genes throughout life may affect gastric function.

Stomach tissue classification using autofluorescence spectroscopy and machine learning.

Determination of stomach tumor location and invasion depth requires delineation of gastric histological structure, which has hitherto been widely accomplished by histochemical staining. In recent years, alternative histochemical evaluation methods have been pursued to accelerate intraoperative diagnosis, often by bypassing the time-consuming step of dyeing. Owing to strong endogenous signals from coenzymes, metabolites, and proteins, autofluorescence spectroscopy is a favorable candidate technique to achieve this aim. We investigated stomach tissue slices and block specimens using a fast fluorescence imaging scanner. To obtain histological information from broad and structureless fluorescence spectra, we analyzed tens of thousands of spectra with multiple machine-learning algorithms and built a tissue classification model trained with dissected gastric tissues. A machine-learning-based spectro-histological model was built based on the autofluorescence spectra measured from stomach tissue samples with delineated and validated histological structures. The scores from a principal components analysis were employed as input features, and prediction accuracy was confirmed to be 92.0%, 90.1%, and 91.4% for mucosa, submucosa, and muscularis propria, respectively. We investigated the tissue samples in both sliced and block forms using a fast fluorescence imaging scanner. We successfully demonstrated differentiation of multiple tissue layers of well-defined specimens with the guidance of a histologist. Our spectro-histology classification model is applicable to histological prediction for both tissue blocks and slices, even though only sliced samples were trained.

Comparison of distribution of verbascoside in normoxic and hypoxic rats

This study established a high performance liquid chromatography(HPLC) method for the simultaneous determination of verbascoside(VB) and its main metabolite caffeic acid(CA) in rat tissue samples. A low-pressure low-oxygen animal experimental chamber was used to simulate the plateau environment for establishing the hypoxic rat model. After intragastric administration of 300 mg·kg~(-1) VB, the normoxic and hypoxic rats were sacrificed for the collection of heart, liver, spleen, lung, kidney, brain, muscle, large intestine, small intestine, and stomach tissue samples at the time points of 30, 60, and 90 min. VB and CA concentrations in each tissue sample were measured by HPLC, and the distribution of VB and CA in normoxic and hypoxic rats was compared. The results showed that after intragastric administration, VB can be rapidly absorbed and distributed into various tissues including brain in both normoxic and hypoxic rats, indicating that VB can pass through the blood-brain barrier. In the gastrointestinal tract, VB was mainly distributed in small intestine, which suggested that the main absorption site of VB was small intestine. A large amount of VB was detected in muscle and lung, and only a small amount in other tissues. CA was detected in other tissues except brain, heart, and muscle. Small intestine had the most abundant CA, followed by stomach, large intestine, and kidney, and only a small amount of CA was detected in the liver, spleen, and lung(<50 ng·mL~(-1)). The results indicated that VB may be mainly absorbed and metabolized in the gastrointestinal tract to produce CA and was possibly excreted through kidney. Compared with normoxic rats, hypoxic rats had reduced and slow distribution of VB and increased ratio of VB concentration in tissue to plasma, which implied that the relative proportion of VB from systemic circulation to tissues was increased in hypoxic rats. This study provides a basis for the application of VB in anti-hypoxia therapy and for the formulation of anti-hypoxia dosing regimens.

Abstract 5680: Hypoxia-responsive and HIF1α-regulated AAA+ ATPase ATAD2 shows high oncogenic potential in stomach cancer

Abstract The burden of stomach cancer incidence and death is a global challenge. Treating stomach cancer remains immensely difficult due to the lack of available markers to detect the disease at the early stages. Therefore, there is an immense demand for finding out an early detection biomarker for stomach cancer. Recently, ATAD2 is found to be overexpressed in stomach cancer with high prognostic significance and recognised as a promising biomarker for stomach cancer. ATAD2 is a unique cancer/testis antigen (CTA) that belongs to both AAA+ ATPase and bromodomain family proteins. However, nothing much is known about the mechanism of ATAD2-mediated stomach carcinogenesis. Using bioinformatics analyses as well as studies with stomach cancer cells and stomach biopsy tissue samples, we address the regulation and function of ATAD2 expression in stomach cancer. We show that ATAD2 is overexpressed in the most common intestinal type of stomach adenocarcinoma, and enhanced expression of ATAD2 is observed in all stages and grades of stomach cancer. Our immunofluorescence microscopy study with stomach biopsy tissues confirms the high expression of ATAD2 both in stomach adenocarcinoma and metastatic stomach tissue samples. Survival analysis indicates that upregulated ATAD2 expression drastically affects the survival of stomach cancer patients. Since H. pylori infection, and hypoxia are the major contributing factors for stomach carcinogenesis; we further study their role in ATAD2-mediated stomach malignancy. An enhanced expression of ATAD2 is observed in H. pylori-infected stomach cancer cells. We identify ATAD2 as a hypoxia-responsive and HIF1α-regulated gene and elucidate that upregulated expression of ATAD2 enhances proliferation and migration of hypoxic stomach cancer cells. We further recognize the protein-protein interaction (PPI) network of ATAD2 with the top-ranked partners like ESR1 (or ERα), NCOA3 (or ACTR/SRC3/AIB1), SUMO2, HDA11 (HDAC11), SPTN2 (SPTBN2), and MYC. Among them, ESR1, MYC, NCOA3, and HDA11 are oncoproteins. Most strikingly, we report the participation of two druggable targets of ATAD2, i.e., AAA+ ATPase and bromodomain in the ATAD2-PPI network; ESR1, SUMO2, SPTN2, and MYC show a preference for bromodomain, whereas NCOA3 and HDA11 prefer ATPase domain of ATAD2. The importance of such findings in terms of targeting bromodomain-PPI and/or ATPase-PPI interfaces to curtail stomach cancer is elucidated. Citation Format: Anasuya Roychowdhury, Aditi Nayak, Sugandh Kumar, Anshuman Dixit, Asima Bhattacharyya. Hypoxia-responsive and HIF1α-regulated AAA+ ATPase ATAD2 shows high oncogenic potential in stomach cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5680.

Preventive and Regenerative Effect of Glutamine and Probiotics on Gastric Mucosa in an Experimental Model of Alcohol-Induced Injury in Male Holtzman Rats

Background: The purpose of this study was to measure the preventive and regenerative effect of glutamine and probiotics induced by alcohol injury in Holtzman rats. Methods: Analytical, experimental and prospective study. The population consisted of 56 male rats between 300 and 350 g, distributed in three experimental phases: Pre-pilot phase PPP (6 rats), Pilot phase PP (10 rats), and Experimental phase EP (40 rats). In the pilot phase, 10 rats were subjected to damage with 8.5% ethanol, which was given intragastrically. The dosage was calculated for 10 rats in two groups: the first with 7.5 mL/kg in 5 rats and the second with 8.5 mL/kg in 5 rats. The experimental phase was performed in 40 rats divided into 6 groups, the negative control group (healthy), positive control group (injured), preventive experimental group (glutamine and glutamine with probiotic) and regenerative experimental group (glutamine and glutamine with probiotic). At the end of each phase, the rats were sacrificed with sodium pentobarbital (Halathal) and a portion of their stomachs was stored in formol. Results: The evaluation of stomach tissue samples (desquamation, erythema, hyperemia) showed that in the preventive phase, glutamine shows effectiveness in comparison to glutamine with probiotic. In the regenerative phase, glutamine and glutamine with probiotic did not show significant differences. Conclusions: Glutamine and probiotics can potentially serve as a therapy for the treatment for gastritis.

Comparative stomach tissue distribution profiles of four major bio-active components of Radix Astragali in normal and gastric ulcer mice

Numerous studies have demonstrated that Radix Astragali can inhibit gastric ulcers in mice. Anhydrous ethanol (0.01 mL/g) administered to mice by intragastric infusion can induce gastric ulcer injury. This study was performed to compare the stomach tissue distribution profiles of four major bioactive constituents of Radix Astragali(calycosin-7-O-β-d-glucoside, calycosin, ononin and formononetin) after oral administration of extract of Radix Astragali (ERA)in normal and gastric ulcer mice. The abundance of Radix Astragali constituents was determined using an ultra-pressure liquid chromatograph with a photodiode array detector (UPLC-PDA), after which histograms were drawn. In comparison with normal mice, the contents of calycosin- 7-O-β-d-glucoside, calycosin, ononin and formononetin in the stomach tissue samples of gastric ulcer mice showed significant differences at the selected time points (P < 0.05).The abundance of each of the four tested constituents in the normal groups was higher than that of the gastric ulcer groups. This study provides an empirical foundation for future studies focused on developing clinical applications of Radix Astragali.

Targeted MicroRNA Profiling in Gastric Cancer with Clinical Assessement.

Although several microRNAs (miRNAs) have been associated with gastric cancer there is still the need for identification of stable and validated biomarkers. The purpose of this study was to determine the alterations of a specific set of miRNA levels in gastric adenocarcinoma tissues to identify and validate gastric cancer-specific miRNAs using paired normal and tumor samples in an independent patient cohort. Gastric adenocarcinoma and normal stomach tissue samples of 20 patients who underwent surgery for gastric cancer were studied. The miRNA expression profiling was performed for eight miRNAs in a total of 40 tissue samples using quantitative reverse transcription polymerase chain reaction (RT-qPCR). Six out of these eight miRNAs, namely, miR-375-3p, hsamiR-129-5p, miR-196a-5p, miR-376c-3p, miR-34c-5p and miR-767-5p, were significantly underexpressed in malignant tissues of our cohort. Furthermore, the expression of miR-662 although not significantly different between normal and tumor tissues, was inversely associated with age (r = −0.440, p = 0.049). The levels of miR-129-3p and miR34c-5p were correlated with an increase in the number of metastatic lymph nodes (r = 0.470, p = 0.036; r = 0.510, p = 0.020), while and miR-376c-3p levels were negatively associated with smoking (p = 0.043). In addition, we found that the variability of miRNA expression in cancerous tissues was lower than that in normal tissues. Alterations in miRNA expression in gastric adenocarcinoma tissues in comparison to healthy tissues of each individual serves for identification of consistent biomarkers that can be used for development of diagnostic tools for gastric cancer.

Nuclear magnetic resonance-based metabolomics revealed metabolite profiles participate in intervention effect of refined moxibustion in gastric ulcer model rats

To investigate the effect of refined moxibustion on expression of gastric mucosal epidermal growth factor receptor (EGFR) and vascular endothelial growth factor (VEGF), and changes of metabolite profiles in gastric ulcer (GU) rats, so as to analyze its mechanism underlying improvement of GU. Male SD rats were randomized into control, model, acupoint moxibustion groups (n=6 per group). The GU model was induced by cold-restraint stress. The ignited refined moxa was applied to bilateral "Liangmen" (ST21) and "Zusanli" (ST36) for 3 cones/acupoint, once daily for 7 days. Then, we employed 1H NMR-based metabolomics approach to analyze the metabolic profiles of serum and stomach tissue samples. The conventional histopathological changes of the gastric mucosa were observed by H.E. stain and the expressions of EGFR and VEGF in the gastric mucosa were detected by immunohistochemistry. Compared to the control group, the expression levels of EGFR and VEGF were significantly increased in the model group (P<0.01, P<0.05), and further notably up-regulated in the acupoint moxibustion group (P<0.001, P<0.01). Results of H.E. staining showed damage of the folds of gastric mucosa, disordered arrangement of the glands, infiltration of inflammatory cells and unclear structure of gastric mucosa in the model group, which was relatively milder in the acupoint moxibustion group. 1H-NMR technical analysis showed that in comparison with the control group, 11 and 11 metabolites in the stomach extract and plasma were increased, 10 in the gastric tissue and 3 in the plasma were decreased in the GU model group; while in comparison with the model group, 17 differently expressed metabolites in the gastric extract and 10 metabolites in the plasma restored to their levels of control group after the acupoint moxibustion intervention. These metabolites participate in 12 metabolic pathways including glycine, serine and threonine metabolism, glutathione metabolism, glycine metabolism, alanine, aspartic acid and glutamic acid metabolism, purine metabolism, glyoxylic acid and digarboxylic acid metabolism, biosynthesis of aminoacyl-tRNA, amino sugar and nucleotide sugar metabolism, cysteine and methionine metabolism, citrate cycle, pyruvate metabolism, and the mutual conversion of pentose and glucuronate，suggesting their involvement in moxibustion-induced improvement of GU. Refined moxibustion at ST21 and ST36 can up-regulate the expression of EGFR and VEGF in the gastric mucosa and lessen gastric mucosal injury, which may be related to its effects in reducing GU-induced metabolic disorders, including sugar, purine, amino acid, and phospholipid metabolism and antioxidant defense system.

Supplementation of Siberian sturgeon (Acipenser baerii) diet with barberry (Berberis vulgaris) fruit extract: Growth performance, hemato-biochemical parameters, digestive enzyme activity, and growth-related gene expression

In this study, barberry fruit extract (BFE) was used as an herbal feed additive at 0 (control), 150, 300, 600, and 750 mg/kg to evaluate growth performance, physiometabolic responses, and gene expression related to growth and appetite in Siberian sturgeon. The fish (N = 150; 29.7 ± 2.2 g) were fed with the experimental diets for 56 days. Upon the completion of the feeding trial, the fish fed with BFE at 750 mg/kg showed higher growth indices than the control fish and the lowest feed conversion ratio (P < 0.05). The highest levels of red blood cell, hemoglobin, and hematocrit were observed in the group fed with 750 mg/kg BFE (P < 0.05). Different dietary levels of BFE had no significant effect on the levels of alkaline phosphatase and alanine aminotransferase in the serum samples (P > 0.05), while a decreasing trend was seen in aspartate aminotransferase, lactate dehydrogenase, glucose, and triglyceride contents by increasing in the dietary levels of BFE. The activities of lipase, amylase, and protease in the BFE-added groups were higher than the control group (P < 0.05). The highest expression of growth hormone gene was recorded in 600 and 750 mg/kg BFE treatments (P < 0.05). The transcription of insulin-like growth factor-I gene was significantly up-regulated in 750 mg/kg BFE treatment compared to others (P < 0.05). Ghrelin gene expression was significantly elevated in both brain and stomach tissue samples by increasing dietary levels of BFE. Overall, dietary BFE markedly at 750 mg/kg exerts growth-promoting effects in Siberian sturgeon.

Preventive and Regenerative Effect of Glutamine and Probiotics on Gastric Mucosa in an Experimental Model of Alcohol-Induced Injury in Male Holtzman Rats

Aims: The purpose of this study was to measure the preventive and regenerative effect of glutamine and probiotics in holtzman rats induced by alcohol injury. Methods: Analytical, experimental and prospective study. The population consisted of 56 male rats between 300 and 350 g, distributed in three phases: Pre-pilot phase PPP (6 rats), Pilot phase PP (10 rats), and Experimental phase EP (40 rats). In the pilot phase, 10 rats were subjected to damage with 8.5% ethanol, which was given intragastrically (3 ml/kg). The dosage was calculated for 10 rats in two groups: the first with 7.5 ml/kg in 5 rats and the second with 8.5 ml/kg in 5 rats. The experimental phase was performed in 40 rats divided into 6 groups, the negative control group (healthy), positive control group (damage), preventive experimental group (glutamine and glutamine with probiotic) and regenerative experimental group (glutamine and glutamine with probiotic). At the end of each phase the rats were sacrificed with sodium pentobarbital (Halathal) and a portion of their stomachs were stored in formol for macroscopic examination and for its corresponding processing and histopathological analysis. Results: The evaluation of stomach tissue samples (desquamation, erythema, hyperemia) showed that in the preventive phase glutamine shows effectiveness in comparison to glutamine with probiotic. In the regenerative phase, glutamine shows better effectiveness than glutamine plus probiotic. Conclusion: Glutamine and probiotics can potentially serve as a therapy for the treatment for gastritis.

S3002 Dysphagia Leading to Endoscopic Diagnosis of Primary Amyloidosis

INTRODUCTION: Amyloidosis is a rare disorder that is caused by the extracellular deposition of abnormal protein fibrils in organs and tissues. Gastrointestinal (GI) involvement in systemic amyloidosis is rare with a reported prevalence of about 3%. We report a case of a 66-year-old female with complaint of dysphagia diagnosed with primary (AL) amyloidosis based on esophagogastroduodenoscopy (EGD) and biopsy. CASE DESCRIPTION/METHODS: A 66-year-old woman with a past medical history of hypertension, diastolic heart failure, history of stroke, and gastroesophageal reflux presented with symptoms of dysphagia accompanied by dysgeusia, anorexia, and unintentional 20 lb weight loss. On physical examination, the patient exhibited mild epigastric tenderness. Laboratory results notable for alkaline phosphatase 440 IU/L, total bilirubin 1.4 mg/dL, hemoglobin 11.7 g/dL, and INR 1.2 with normal aspartate aminotransferase and alanine aminotransferase, and negative hepatitis serologies. The patient underwent EGD with multiple large non-bleeding angioectasias found in the stomach, which were treated with argon plasma coagulation (APC) prophylactically. Patchy, moderately erythematous mucosa without bleeding was found in the stomach and was biopsied. No gross endoscopic esophageal abnormality was found to explain the patient's dysphagia. Four biopsies were obtained in the proximal and distal esophagus. Random gastric biopsies stained with Congo red demonstrated amyloidosis involving oxyntic mucosa with mild chronic inactive gastritis. Distal esophageal biopsies stained with Congo red demonstrated deposits suspicious for amyloid. DISCUSSION: GI-related symptoms in AL amyloidosis is reported to be 8–60%. However, the prevalence of gastrointestinal amyloidosis with both symptoms and biopsy-proven involvement of tissue is about 3%. Common symptoms of GI amyloidosis include weight loss, diarrhea, abdominal pain, and varying degrees of bleeding. The above patient presented with dysphagia, which is a rarely reported symptom of GI amyloidosis. The gold standard for diagnosing amyloidosis is tissue biopsy with Congo red stain demonstrating green birefringence under polarized light. Both the esophageal and stomach tissue samples demonstrated amyloid deposition. The dysphagia experienced by the patient was likely due to neuromuscular infiltration causing dysmotility. This case demonstrates the unusual presentation of dysphagia leading to the diagnosis of AL amyloidosis with rare biopsy proven infiltration of the esophagus and stomach.Figure 1.: Stomach biopsy sample stained with Congo red demonstrating apple green birefringence under polarized light.Figure 2.: Esophagus biopsy sample stained with Congo red demonstrating apple green birefringence under polarized light.Figure 3.: Stomach biopsy sample stained with Congo red demonstrating congofilic deposits.

Immunohistochemical Studies of Age-Related Changes in Cell Proliferation and Angiogenesis during the Healing of Acetic Acid-Induced Gastric Ulcers in Rats.

Cell proliferation and angiogenesis are of utmost importance for healing to take place. The KI67 and EGFR proteins are markers of cell proliferation, while CD31 and factor VIII are markers of angiogenesis. To elucidate the mechanism responsible for delayed healing of the gastric injury in old age, we analyzed the expression of these markers in rats of different months during the healing of an acetic acid-induced gastric ulcer. Male Wistar rats (aged 3, 6, 12, and 18 months) divided into four groups, according to their ages, formed the experimental animals. Stomach tissue samples were collected on days 3, 7, 14, and 21 after induction for assessment of ulcer healing. The area of gastric mucosa healed was inversely proportional to age. The expression of markers of proliferation (KI67 and EGFR) and angiogenesis (factor VIII and CD31) decreased significantly (p < 0.05) in older rats when compared with younger ones (3 months > six months > 12 months > 18 months) on days 7, 14, and 21 after induction of gastric ulcer. This study revealed that the slower gastric ulcer healing rate in older rats might be due to reduced epithelial cell proliferation and angiogenic activities.

GHSR DNA hypermethylation is a new epigenetic biomarker for gastric adenocarcinoma and beyond.

Aberrations of DNA methylation are early events in the development of tumors. In this study, we investigated the DNA methylation status of growth hormone secretagogue receptor (GHSR), a promising pan-cancer biomarker, in gastric cancer (GC). Initially, data sets from DNA methylation and gene expression studies available at Gene Expression Omnibus (GEO) were analyzed. Confirmation was done on primary tumor specimens and adjacent normal stomach tissue samples. Both analyses showed significant hypermethylation of GHSR. For further validation, The Cancer Genome Atlas data on stomach cancer was used. A receiver operating characteristic curve analysis yielded an area under the curve value of 0.85, corroborating its usefulness as a diagnostic marker. A genome-wide comethylation analysis revealed several correlated genes. CREB1 was found to act as an upstream regulator of this gene network. Furthermore, GHSR methylation was found to be a biomarker in several other tumor entities, namely cancers of the bladder, endometrium, esophagus, head and neck, liver, thyroid, kidney, and ovary. Our findings along with previous reports on other types of cancer suggest a high potential of GHSR gene methylation as a pan-cancer biomarker, which could be considered for liquid biopsy applications.