Intestinal Disease Research Articles

Microbial catabolism of coffee pulp (poly)phenols during in vitro colonic fermentation

Coffee pulp is a by-product characterized by its richness in phenolic compounds. This study examined the catabolism of (poly)phenols in digested coffee pulp flour (CPF) and extract (CPE) during in vitro colonic fermentation. After a simulated gastrointestinal digestion, samples were fermented using human microbiota and (poly)phenol transformations were analyzed by UHPLC-ESI-MS/MS. Digested CPF and CPE contained high amounts of phenolic acids, notably 3′,4′-dihydroxycinnamic (99.7–240.1 μmol 100 g−1) and 3,4-dihydroxybenzoic acid (174.1–491.4 μmol 100 g−1). During the in vitro fecal fermentation, phenylpropanoic acids (1.5- to 2.6-fold), phenyl-γ-valerolactones (1.3- to 23-fold), phenylvaleric acids (1.1- to 2-fold) and benzene derivatives (1.5-fold) increased; while benzoic and cinnamic acids, cinnamoylquinic derivatives, flavonols, benzaldehydes and diphenylpropan-2-ols decreased. The (poly)phenols in CPF were catabolized more slowly than in CPE, suggesting protection of the fibrous matrix against phenolic degradation. Coffee pulp may be a promising food ingredient rich in (poly)phenols contributing to the prevention of intestinal diseases.

324 Use of organoids as in vitro model to study intestinal function

Abstract Organoids are tridimensional structures cultured in the lab from stem cells. Organoids have been developed for many different organs, particularly those with Lgr5+ stem cells. The organs of the digestive system belong to this group of organs and to date, organoids have been established from all the digestive tract tissues and organs including liver, gall bladder and pancreas. Although most protocols to generate organoids have focused on human and mouse organs, organoids from domestic and production animals like dogs, cats, rabbits, pigs, chickens, turkeys, and horses are actively used in research. In this presentation, the principles of developing intestinal organoids will be covered, along with a summary of how they have been used to address intestinal function and disease in our lab and by others. In our lab, we use swine enteroids (intestinal organoids) to study the intestinal epithelial responses to nutrients and pathogens. We have explored differential responses of the intestinal epithelium to organic and inorganic sources of dietary zinc, and we are currently investigating the mechanism of pathogenesis of Lawsonia intracellularis, the causative agent of porcine intestinal adenopathy. We will also discuss methods for modeling intestinal physiology using enteroids, and the advantages and challenges of working with swine enteroids.

Synergistic effect and mechanism of monoacylglycerol lipase inhibitor and Icaritin in the treatment of ulcerative colitis

Ulcerative colitis (UC) is a chronic, relapsing nonspecific intestinal inflammatory disease. It is difficult for a single drug to treat UC effectively and maintain long-term efficacy. There is an urgent need to find new drugs and treatment strategies. MAGL11 is a new kind of single acylglycerol lipase (MAGL) inhibitor. Icaritin (Y003) is the major metabolite of icariin in vivo. Several studies have confirmed the role of MAGL inhibitors and icariin in anti-inflammatory and regulation of intestinal stability. Therefore, this study adopted a new strategy of combining MAGL inhibitor with Icaritin to further explore the role and mechanism of drugs in the treatment of UC. Enzyme-linked immunosorbent assay (ELISA), hematoxylin-eosin staining (HE), immunohistochemical (IHC) and Western blot were used to detect the synergistic protective effects of MAGL11 and Y003 on intestinal pathological injury, intestinal mucosal permeability and inflammation in UC mice. 16S rDNA sequencing was used to detect the synergistic effect of MAGL11 and Y003 on gut microbiota. The effects of MAGL11 and Y003 combined therapy on serum and fecal metabolism of UC mice were analyzed by untargeted metabolomics. Proteomics method was applied to investigate the molecular mechanisms underlying MAGL11 and Y003 synergy in the treatment of UC. The results showed that MAGL11 and Y003 could synergistically improve the clinical symptoms, reduce intestinal inflammation and pathological damage, and improve intestinal mucosal permeability in UC mice. The mechanism study found that MAGL11 and Y003 could synergistically inhibit Toll-like receptors 4 (TLR4) / Myeloid differentiation primary response gene (Myd88)/Nuclear factor kappa-B (NF-κB) pathway and further regulate gut microbiota imbalance and metabolic disorders to treat UC.

30 Microbiota and Companion Animal Health

Abstract The intestinal microbiota has been shown to have an important role in health and disease of animals. It is a complex system that depends on interactions between the luminal environment and specific bacterial taxa and their function (e.g., production of short-chain fatty acids, conversion of bile acids). Dietary substrates are an important factor for microbial metabolism and supplements have the potential to help in maintaining intestinal health. Major changes in the microbiota and their loss of function (dysbiosis) are mainly associated with antibiotic treatment and in chronic intestinal disease likely due to chronic remodeling of the epithelium. Therefore, often these changes persist long-term. Chronic enteropathies (CE) occur very commonly in dogs and cats, and are frustrating to treat due to lack of specific markers. The pathophysiology is complex and includes changes in the immune system (e.g., inflammation), intestinal function, and the intestinal microbiome. The current classification based on treatment response (i.e., food- vs. steroid-responsive) is currently being questioned, as new research findings indicate that CE is a syndrome with many overlapping unspecific features, often with chronic irreversible changes in intestinal function and dysbiosis. The presentation will summarize novel research about the pathophysiology and novel concepts in multi-modal therapy to these conditions. This program will help better understand the role of the intestinal microbiota in health and disease (with focus on acute diarrhea and chronic intestinal inflammation). New diagnostic methods for evaluation of the intestinal microbiota will be covered. Finally, this lecture will review how to best manage the clinical signs of acute and chronic diarrhea. This includes new evidence of the negative impact of antibiotics on the microbiome and the critical role nutrition can play in managing these patients.

The Burden of Psychiatric Manifestations in Inflammatory Bowel Diseases: A Systematic Review With Meta-analysis.

Psychiatric disorders in patients with inflammatory bowel disease (IBD) represent a significant but uncertain facet of the disease, with unsolved questions regarding their overall magnitude, their impact on intestinal disease, and the whole burden of psychiatric manifestations. This systematic review summarizes the evidence on the prevalence and impact of psychiatric disorders, including depression, anxiety, bipolar disorder (BD), and schizophrenia, among patients with IBD. A systematic search across PubMed/MEDLINE, Embase, and Scopus databases from January 2010 to January 2023 was performed to identify relevant studies. The focus was on studies exploring the prevalence of specific psychiatric disorders in IBD patients compared to the general population and that reported specific outcome measures. A subsequent meta-analysis (MA) assessed the strength of the association between IBD and these psychiatric disorders, with data reliability ensured through rigorous extraction and quality assessment. Out of 3,209 articles, 193 met the inclusion criteria and only 26 provided complete data for comprehensive analysis. These studies showed a significantly higher overall prevalence of psychiatric comorbidities in IBD patients compared to the general population. The MA showed a significant association between IBD and depression (pooled OR 1.42, 95% CI = 1.33-1.52, P < .0001) and anxiety (pooled OR 1.3, 95% CI = 1.22-1.44, P < .0001). The association between IBD and BD was significant (pooled OR 1.64, 95% CI = 1.20-2.24, P < .0001) but showed considerable heterogeneity (I2 = 94.01%). Only 3 studies examined the association between schizophrenia and IBD, providing widely heterogeneous results, with an inconclusive OR, estimated at 0.93 (95% CI = 0.62-1.39, P = .73). This MA highlights the high prevalence of psychiatric disorders, particularly depression and anxiety, in IBD patients, which exceeds rates in the general population. BD in IBD is proving to be an important but under-researched area. The sparse and contradictory data on schizophrenia requires further investigation. These findings highlight the need for better understanding, early detection, and tailored mental health interventions in the management of IBD to significantly improve patients' quality of life.

Critical Suitability Evaluation of Caco-2 Cells for Gut-on-a-Chip.

Currently, culturing Caco-2 cells in a Gut-on-a-chip (GOC) is well-accepted for developing intestinal disease models and drug screening. However, Caco-2 cells were found to overexpress surface proteins (e.g., P-gp) compared with the normal intestinal epithelial cells in vivo. To critically evaluate the challenge and suitability of Caco-2 cells, a GOC integrated with a carcinoembryonic antigen (CEA) biosensor was developed. This three-electrode system electrochemical sensor detects CEA by antigen-antibody specific binding, and it exhibits high selectivity, excellent stability, and good reproducibility. Under dynamic culturing in the GOC, Caco-2 cells exhibited an intestinal villus-like structure and maintained tissue barrier integrity. Meanwhile, CEA was discovered to be secreted from 0 to 0.22 ng/mL during the 10-day culturing of Caco-2 cells. Especially, CEA secretion increased significantly with the differentiation of Caco-2 cells after 6 days of culturing. The sustained high-level CEA secretion may induce cells to avoid apoptotic stimuli, which faithfully reflects the efficacy of a new drug and the mechanism of intestinal disease. Different kinds of cell types (e.g., intestinal primary cells, stem cell-induced differentiation) in the GOC should be attempted for drug screening in the future.

Exploration of JI Laixi's academic thought of acupuncture and moxibustion in clinical practice

The paper summarizes Professor JI Laixi's academic thought on acupuncture and moxibustion. Professor JI Laixi inherits and carries forward the theory and technical system of Shanxi "new nine needles", deepens its academic connotation, and promotes clinical practice. He advocates the integration of Chinese and western medicine, and the inclusion of multiple disciplines, e.g. acupuncture and moxibustion of Chinese medicine and modern anatomy. He takes the lead in proposing a new approach to the treatment of meridian diseases of limbs, "treating the neck region for the diseases of head, treating the abdominal region for the diseases of the lumbar region, and treating the lumbar region for the disease of knees". He proposes the acupoint prescription being standardized and simplified, explores the system of acupoint prescription and establishes the prescriptions for gastric diseases, intestinal diseases and antihypertension.

Itaconic Acid Alleviates Perfluorooctanoic Acid-Induced Oxidative Stress and Intestinal Damage by Regulating the Keap1/Nrf2/Ho-1 Pathway and Reshaping the Gut Microbiota.

Itaconic acid (IA) is recognized for its potential application in treating intestinal diseases owing to the anti-inflammatory and antioxidant properties. Perfluorooctanoic acid (PFOA) can accumulate in animals and result in oxidative and inflammatory damages to multi-tissue and organ, particularly in the intestinal tract. This study aimed to explore whether IA could mitigate intestinal damage induced by PFOA exposure in laying hens and elucidate its potential underlying mechanisms. The results showed that IA improved the antioxidant capacity of laying hens and alleviated the oxidative damage induced by PFOA, as evidenced by the elevated activities of T-SOD, GSH-Px, and CAT, and the decreased MDA content in both the jejunum and serum. Furthermore, IA improved the intestinal morphological and structural integrity, notably attenuating PFOA-induced villus shedding, length reduction, and microvillus thinning. IA also upregulated the mRNA expression of ZO-1, Occludin, Claudin-1, and Mucin-2 in the jejunum, thereby restoring intestinal barrier function. Compared with the PF group, IA supplementation downregulated the gene expression of Keap1 and upregulated the HO-1, NQO1, SOD1, and GPX1 expression in the jejunum. Meanwhile, the PF + IA group exhibited lower expressions of inflammation-related genes (NF-κB, IL-1β, IFN-γ, TNF-α, and IL-6) compared to the PF group. Moreover, IA reversed the PFOA-induced imbalance in gut microbiota by reducing the harmful bacteria such as Escherichia-Shigella, Clostridium innocuum, and Ruminococcus torques, while increasing the abundance of beneficial bacteria like Lactobacillus. Correlation analysis further revealed a significant association between gut microbes, inflammatory factors, and the Keap1/Nrf2/HO-1 pathway expression. In conclusion, dietary IA supplementation could alleviate the oxidative and inflammatory damage caused by PFOA exposure in the intestinal tract by reshaping the intestinal microbiota, modulating the Keap1/Nrf2/HO-1 pathway and reducing oxidative stress and inflammatory response, thereby promoting intestinal homeostasis.

Engineering probiotic Escherichia coli Nissle 1917 to block transfer of multiple antibiotic resistance genes by exploiting a type I CRISPR-Cas system.

To reduce the development of antibiotic resistance, probiotics have been considered as a substitute for antibiotics. However, probiotics themselves are reservoirs of antibiotic resistance genes (ARGs). This study introduces a new strategy for limiting the spread of ARGs by engineering the typical probiotic strain Escherichia coli Nissle 1917 (EcN), which has been used for treating intestinal diseases and developed as living therapeutics. We also demonstrate that the type I CRISPR-Cas system imposes a lower growth burden than the type II CRISPR-Cas system, highlighting its promising clinical application potential. Our work not only provides a new strategy for restricting the transfer of ARGs while using probiotics but also enriches the genetic engineering toolbox of EcN, paving the way for the safe use and development of probiotics as living therapeutics.

Penerapan Metode Teorema Bayes untuk Memprediksi Penyakit Tifus

Typhoid is an acute febrile condition caused by infection with Salmonella enterica bacteria, especially the Salmonella typhi variant. Typhoid fever or what we usually know as typhoid fever. However, this disease can also be caused by other types such as Salmonella paratyphi A, Salmonella typhi B, and Salmonella paratyphi C. Typoid fever or typhus abdominalis is an acute infectious disease of the small intestine with symptoms of fever for one week or more accompanied by disorders of the intestinal tract. digestion and with or without impaired consciousness. Bayes' theorem is a theory of probability conditions that takes into account the possibility of an event (hypothesis) depending on other events (evidence). Future events can be predicted if previous events have occurred. Bayes' theorem is a mathematical equation used in probability and statistics to calculate conditional probabilities. In other words, it is used to calculate the probability of an event based on its relationship to other events. Based on the weight value given by the expert to each patient's typhoid symptom data, from the results of the analysis carried out with the diagnosis results from the consultation, the symptoms are High fever (lasting up to two weeks), Headache, Chills, Skin rash, Muscle and joint pain, Extreme fatigue, Dry cough, Confusion or delirium, Nausea and vomiting, Swollen spleen, Abdominal pain with predicted results for Epidemic Typhus with a value of 76.26%.

The association between erythema nodosum and pyoderma gangrenosum and pediatric inflammatory bowel disease.

The objectives of this study is to estimate rates and identify factors associated with erythema nodosum (EN) and pyoderma gangrenosum (PG) in pediatric patients with inflammatory bowel disease (IBD). This cohort study examined longitudinal visits of patients aged ≤ 21 years from the ImproveCareNow (ICN) registry. We evaluated the association of factors at the patient-level (demographics and IBD diagnosis age) and visit-level (IBD severity scores, markers and phenotypes, comorbidities, and treatment) with the presence of EN and PG, using longitudinal logistic regression models adjusted for time and within-patient clustering. A total of 285,913 visits from 32,497 patients aged ≤ 21 years from the ICN registry were analyzed. The occurrence of EN was 1.57% (95% confidence interval [95% CI]: 1.43%-1.71%) and the occurrence of PG was 0.90% (95% CI: 0.80%-1.00%). Co-occurrence of EN and PG was reported in 0.30% (95% CI: 0.25%-0.37%) patients. Both EN and PG were associated (p < 0.0001) with worse intestinal disease, lower remission, higher inflammatory markers, and extraintestinal manifestations (EIMs) arthritis and uveitis. EN and PG were associated with increased disease severity and other noncutaneous EIMs (arthritis and uveitis). A small subset of patients had developed both EN and PG.

Prevalence and genomic characteristics of becAB-carrying Clostridium perfringens strains

Clostridium perfringens, as a foodborne pathogen, can cause various intestinal diseases in both humans and animals according to its repertoire of toxins. In recent years, a multitude of studies have highlighted its threat to infants and young children. C. perfringens carries numerous toxins, with the newly identified BEC toxin confirmed as the second toxin to cause diarrheal illness, after CPE. However, the global dissemination of C. perfringens strains carrying becAB genes, which encode BEC toxins, has not been extensively studied. Following epidemiological surveillance of the prevalence of C. perfringens from different sources in various provinces of China, we identified two becAB-carrying strains and one strain carrying a sequence similar to becAB from distinct provinces and sources. When combined with genomic analysis of other becAB-carrying C. perfringens strains from public databases, we found that becAB was present in strains from different lineages. Our analysis of the plasmid and genetic environment corroborates previous findings on becAB-carrying strains, confirming that it currently achieves horizontal transmission through one type of evolutionarily conserved Pcp plasmid. This study provides a comprehensive analysis of the prevalence and transmission patterns of the newly emerged toxin gene locus, becAB, in C. perfringens. Despite the relatively low identification rate of becAB-carrying strains, their potential impact requires ongoing surveillance and investigation of their features, particularly their antimicrobial resistance.

A planar mounted SQUID full-tensor module for magnetoenterogram denoising detection

Magntoenterogram (MENG) is a functional imaging method with great clinical potential for non-invasive diagnosis of intestinal diseases. In measurements of MENG, the cardiac artifacts are a strong biological source of interference. Based on wire-wound SQUID gradiometers, we constructed a planar mounted full-tensor compensation module for MENG system, which can obtain cardiac artifacts while suppressing ambient noise, so as to denoise MENG signals without increasing external ECG recording channels. The cardiac artifacts are regressed out from the full-tensor compensation module outputs, which can denoise the MENG measurements. Using this module, the SNR is about 70 dB better than the raw data. At last, 36-point array MENG signals were detected successfully and the contour map of jejunal slow-wave frequency (SWF) was consistent with the jejunal area (one part of the small intestine). This method will save costs and avoid the new electromagnetic interferences introduced by ECG equipment. It will lay a good foundation for the rapid clinical application of the MENG system.

Design and Synthesis of an Azo Reductase Responsive Flavonol-Indomethacin Hybrid Used for the Diagnosis and Treatment of Colitis.

Human intestinal bacteria are the primary producers of azo reductase, and the content of azo reductase is closely associated with various intestinal diseases, including ulcerative colitis (UC). The rapid detection of changes in azo reductase levels is crucial for diagnosing and promptly intervening in UC. In this study, a therapeutic agent, FAI, specifically targeting UC, was designed and synthesized. This agent was developed by linking the anti-inflammatory drug indomethacin to flavonols with antioxidant activity via an azo bond (off-on). Breakage of the azo bond breaks results in the release of both fluorophores and drugs, achieving targeted tracing and integrated treatment effects. In vivo and in vitro fluorescence imaging experiments were used to demonstrate the potential of FAI in the diagnosis of UC, together with synergistic therapeutic effects through the release of both fluorophores and anti-inflammatory agents. Therefore, this diagnostic agent shows promise as a potential tool for diagnosing and treating UC.

The relationship between mental health problems and risk of infectious diseases: A Mendelian randomization analysis.

The causal effects of mental health problems on the risk of infectious diseases remain vague. Investigating them via observational study is challenging as it presents possible confounding factors. Therefore, the objective of this study was to utilize Mendelian randomization (MR) techniques to evaluate the causal relationship between mental health problems and the risk of infectious diseases. Multivariable MR analyses were performed using genome-wide association data for sleep disorders (N = 216,700), depression (N = 500,199), anxiety (N = 290,361), nervous feelings (N = 450,700), unspecified mental disorder (N = 218,792), pneumonia (N = 486,484), skin and subcutaneous tissue infection (SSTI; N = 218,792), intestinal infectious diseases (IIDs; N = 218,792), urinary tract infection (N = 463,010), and central nervous system (CNS) infections (N = 218,792) among individuals of European ancestry. Independent genetic variants significantly (P < 10-8) associated with each exposure were considered instruments. The primary analysis used an inverse variance-weighted method, followed by a series of sensitivity analyses. Genetically predicted sleep disorders were associated with an increased risk of SSTI (odds ratio [OR], 1.29 [95% confidence interval (CI), 1.05-1.59]; P = .017). Genetically predicted depression was linked with an increased risk of CNS infections (OR, 1.59 [95% CI, 1.00-2.53]; P = .049) and SSTI (1.24 [95% CI, 1.03-1.49]; P = .024). Genetically predicted anxiety was associated with IIDs (OR, 1.19 [95% CI, 1.03-1.37]; P = .017) and SSTI (OR, 1.21 [95% CI, 1.02-1.43]; P = .029). There was no significant causal evidence for genetic prediction of nervous feelings and unspecified mental disorders in IIDs, CNS infections, SSTI, pneumonia, or urinary tract infection. Sensitivity analyses showed that the above causal association estimates were robust. In this MR study, we demonstrated a causal relationship between sleep disorders, depression, anxiety, and the risk of infectious diseases. However, no evidence was found to support causality between nervous feelings, unspecified mental disorders, and the risk of infectious diseases.

Infectious Intestinal Disease Survey During COVID-19: Additional Analysis on Association of Behaviours With Illness

During the COVID-19 pandemic businesses and individuals changed hygiene, eating, travelling and social behaviours. These changes have the potential to play an important role in the likelihood of people contracting diseases, including Infectious Intestinal Disease (IID). Six waves of a nationally representative survey were carried out to gather information about IID among the general population during this period. Questions were also asked about the behaviours of those who got IID and of a subset of individuals without IID. Separate surveys were run for adults and children (with parents responding on behalf of their children). Waves 1 to 4 were run for both adults and children, wave 5 was just for adults and wave 6 was just for children. Logistic regression models were used to examine the association between the behaviours and IID, for individual waves and for a combined all-waves analysis, for adults and children separately. Adults eating food from takeaways or street food vendors was the single behaviour positively associated with IID across all five waves, whereas two other behaviours were positively associated with IID in four of the five waves: buying ready to eat food outside work/school; and eating food from work/school canteen. All three of these behaviours were among those selected in the model using data from all five waves. For the children’s analysis there were few behaviours selected in any of the individual models and those that were selected only featured in one of the five models. For the children’s all-waves model the strongest effects were: use of public transport more than weekly; anybody leaving the house; and children eating ready to eat food outside of schools. In the hypothetical scenario of entirely removing eating food from takeaways or street food vendors for adults, the expected reduction in IID cases is 9-24%.

Survey of Infectious Intestinal Disease in the UK: Effect of COVID-19 Response and Associated Measures on IID in the UK

Survey of Infectious Intestinal Disease in the UK: Effect of COVID-19 Response and Associated Measures on IID in the UK

Advances in Research on the Relationship between Diet, Gut Microbes, and Health

In recent years, as research on gut microbiota has deepened, the relationship between diet, gut microbiota, and health has garnered widespread attention. As the largest micro-ecosystem in the human body, gut microbiota not only participates in physiological processes such as food digestion and nutrient absorption but also exerts a profound impact on human health through its metabolic products and interactions with the host. This paper reviews the impact of diet on gut microbiota, and the relationship between gut microbiota and health, aiming to provide new strategies and ideas for promoting human health. Firstly, the article summarizes the effects of different dietary components and dietary patterns on the structure and function of gut microbiota. Then, it analyzes the role of gut microbiota in maintaining human health and preventing diseases and further explores the relationship between gut microbiota and intestinal diseases and metabolic syndrome. The analysis reveals that diet affects human health by influencing the structure and function of gut microbiota, while gut microbiota, as a complex ecosystem within the human body, plays an indispensable role in maintaining human health.

Pediatric Chronic Intestinal Failure: Something Moving?

Pediatric chronic intestinal failure (PIF) is a rare and heterogeneous condition characterized by the inability of the patient's intestine to adequately absorb the required fluids and/or nutrients for growth and homeostasis. As a result, patients will become dependent on home parenteral nutrition (HPN). A MEDLINE search was performed in May 2024 with keywords "intestinal failure", "parenteral nutrition" and "pediatric". Different underlying conditions which may result in PIF include short bowel syndrome, intestinal neuromuscular motility disorders and congenital enteropathies. Most common complications associated with HPN are catheter-related bloodstream infections, catheter-related thrombosis, intestinal failure-associated liver disease, small intestinal bacterial overgrowth, metabolic bone disease and renal impairment. Treatment for children with PIF has markedly improved with a great reduction in morbidity and mortality. Centralization of care in specialist centers and international collaboration between centers is paramount to further improve care for this vulnerable patient group. A recently promising medical therapy has become available for children with short bowel syndrome which includes glucagon-like peptide 2, a naturally occurring hormone which is known to delay gastric emptying and induce epithelial proliferation. Despite advances in curative and supportive treatment, further research is necessary to improve nutritional, pharmacological and surgical care and prevention of complications associated with parenteral nutrition use.

Novel Role of the ALPI Gene Associated with Constipation Caused by Complement Component 3 Deficiency.

Complement component 3 (C3) deficiency has recently been reported as one of the novel causes of constipation. To identify a unique gene specific to constipation caused by C3 deficiency, the total RNA extracted from the mid colon of C3 knockout (C3 KO) mice was hybridized to oligonucleotide microarrays, and the function of the candidate gene was verified in in vitro and in vivo models. C3 KO mice used for microarrays showed definite phenotypes of constipation. Overall, compared to the wild type (WT), 1237 genes were upregulated, and 1292 genes were downregulated in the C3 KO mice. Of these, the major genes included were lysine (K)-specific demethylase 5D (KDM5D), olfactory receptor 870 (Olfr870), pancreatic lipase (PNLIP), and alkaline phosphatase intestinal (ALPI). Specifically, the ALPI gene was selected as a novel gene candidate based on alterations during loperamide (Lop)-induced constipation and intestinal bowel disease (IBD). The upregulation of ALPI expression treated with acetate recovered the expression level of mucin-related genes in primary epithelial cells of C3 KO mice as well as most phenotypes of constipation in C3 KO mice. These results indicate that ALPI plays an important role as the novel gene associated with C3 deficiency-induced constipation.