Gastroenterology and Hepatology from Bed to Bench

Indirect immunofluorescence on HEp-2 cell-line (HEp-2-IIF) remains the “gold standard” method for detection of antinuclear antibodies (ANA) (1). ANA is an operative definition, showing the possibility of autoantibodies (Aab) to bind nucleus and cytoplasmic antigen. One of the major examples is represent by anti-mitochondrial antibodies (AMA), which target proteins of the inner and outer mitochondrial membranes, located in the cytoplasm. In routinely lab-life the AC-21 pattern and the presence of related AMA can be confirm by commercial line-immunoblotting, ELISA, CLIA/FEIA assays and the standard IIF on rodent kidney/stomach/liver tissue sections. Several studies showed the use of other commercial assays and home made tests to detect AMA as immunoprecipitation and immunoblotting. However the use of IP or IP-WB in a routinely laboratory is difficult to apply, because of numerous cases to process and the related troubles.

The routinely lab experience also teach that commercial kits can not always detected and define specific AMA. Where find confirmation of AC-21 pattern if line-immunoblot and other routinely methods (ELISA, CLIA/FEIA assays) fail? We review AC-21 AMA-like sera from our patinets (year 2022) and purpose a revised diagnostic algorithm based on the combined use of IIF on Hep-2 cells, line immunoblot and IIF on rodent tissue as third line method. We showed that the use of IFI on rodent tissues became diriment to confirm AMAs presence, expecially when second level assay failed.

Aim: A systematic review and meta-analysis were performed to investigate posterior tibial nerve electrical stimulation application methods in patients with chronic constipation.

Background: Posterior tibial nerve electrical stimulation is a management procedure for chronic constipation.

Methods: A comprehensive search was conducted on Ovid, PubMed, Scopus, ProQuest, Web of Science, and The Cochrane Library based on the PICO formation of the study. All randomized controlled trials and quasi-experimental studies in which patients with chronic constipation were treated with transcutaneous tibial nerve stimulation (TTNS) or percutaneous tibial nerve stimulation (PTNS) were included in this study. Two independent reviewers screened all titles, abstracts, and full texts. The selected studies' quality was assessed critically using the Joanna Briggs Institute checklists. The data synthesis was conducted using Review Manager software.

Results: Out of 1016 records, 11 studies were included in this study. The results showed that TTNS was effective in improving constipation symptoms (SMD: -1.52, CI 95%: -2.81 to -0.22, p< 0.0001) and reducing defecation time of patients with chronic constipation (SMD: -0.86, CI 95%: -1.60 to -0.13, p= 0.17). Additionally, PTNS was found to improve the quality of life of these patients (SMD: -1.32, CI 95%: -2.05 to -0.59, p< 0.00001).

Conclusion: Both TTNS and PTNS can be effective interventions for chronic constipation. To suggest a definitive and standard treatment plan, further research is needed to determine optimal parameters for TTNS and PTNS applications.

Weight gain post-liver transplant can lead to adverse patient outcomes in the post-transplant period. Pharmacotherapy and other measures can be utilised to reduce the burden and occurrence of weight gain in this population. We explored the mechanism of action, safety, and efficacy of these medications, specifically GLP-1 receptor agonists and metformin, focusing on liver transplant patients.

This scoping review was conducted in line with the scoping review structure as outlined by the PRISMA guidelines.  Metformin and GLP-1 receptor agonists have been observed to be safe and effective in liver transplant patients. Experimental models have found liver-centric weight loss mechanisms in this drug cohort.  There is a paucity of evidence about the use of antihyperglycemics in a post-transplant population for weight loss purposes. However, some small studies have shown strong safety and efficacy data. The evidence in relation to using these medications in patients with metabolic syndrome for weight loss warrants further study in a transplant population. 

Aim: Clomipramine (tricyclic antidepressant), Risperidone (a non-typical antidepressant), and Escitalopram (selective serotonin reuptake inhibitor antidepressant) might be good candidates for investigating the anti-colitis activity.

Background: The incidence of depression with ulcerative colitis in patients has led to the use of antidepressants in their treatment. In addition to the antidepressant effect of these drugs, anti-inflammatory effects have also been reported.

Methods: In this study, 36 rats were used 2 ml of 3% acetic acid solution rectally to show the colitis. Then, Clomipramine (25 mg/kg), Escitalopram (10 mg/kg), Prednisolone (5 mg/kg), Risperidone (2 mg/kg), and normal saline as the control was administered orally for six days. The levels of Tumor Necrosis factor-alpha (TNF-α), Interleukin-6 (IL-6), and myeloperoxidase (MPO) were measured by Enzyme-linked immune sorbent assay (ELISA), and changes in the tissue pathology were investigated.

Results: IL-6 level was significantly reduced after the administration of clomipramine and Prednisolone (p=0.025). Risperidone has significantly reduced MPO activity in colonic tissue (P=0.006). We did find no statistical decrease in MPO activity and TNF-α and IL-6 levels after consumption of Escitalopram (p>0.05).

Conclusion: Clomipramine showed the best anti-inflammatory effect compared to Escitalopram and Risperidone. Therefore, clomipramine showed the best relieving effect on inflammation of ulcerative colitis in rats.

Aim: We aim to investigate the relation of these two circular RNAs with clinicopathological data of colorectal cancer patients.

Background: Circular RNAs (circRNAs) are implicated in the progression and development of Colorectal cancer (CRC). CircRNAs have been recognized as diagnostic and prognostic biomarkers and opening up a new window in order to comprehend the molecular basis of CRC. Given the significance of circRNAs and the G protein subunit b1 (GNB1) gene in malignancies, the goal of the current investigation was to determine the expression levels of GNB1 derivative circular RNAs circGNB1 (hsa_circ_0009361 and hsa_circ_0009362) in CRC and nearby control tissues.

Methods: The expression levels of the GNB1 derivative circular RNAs hsa_circ_0009361 and hsa_circ_0009362 in 45 CRC tissues and nearby control tissues were evaluated using the quantitative real-time PCR (qRT-PCR) method. All other data analyzed by GraphPad Prism software (La Jolla, CA).

 Furthermore, we analyzed the diagnostic power of the mentioned circRNAs by plotting the receiver operating characteristic (ROC) curve. The association between the expression levels of hsa_circ_0009361 and hsa_circ_0009362 was evaluated using correlation analysis.

Result: Our results revealed that the expression levels of hsa_circ_0009361 and hsa_circ_0009362 were significantly down-regulated in CRC tissues compared to the adjacent control group. Analysis of patients’clinicopathological features shows that, expressions hsa_circ_0009361 and hsa_circ_0009362 were differently related with lymph vascular invasion (P < 0.001). ROC curve results have shown that these circRNAs are good candidate diagnostic biomarkers in CRCs. Pearson’s correlation test showed a positive correlation between hsa_circ_0009361 and hsa_circ_0009362 expression levels (P < 0.0001).

Conclusion: These results revealed that hsa_circ_0009361 and hsa_circ_0009362 expression levels may be used as possible diagnostic biomarkers for CRC.

Aim: The potency of Adenovector expressing Mda7-tLyp1 (Ad-Mda7-tLyp1) for death induction was evaluated on the breast (MCF7), liver (HepG2), and gastric (MKN45) cancer cell lines.

Background: Mda-7 could be a possible complementary to traditional cancer therapy, and tethering to tumor-homing peptides (THPs) might improve its therapeutic efficacy.

Methods: After the preparation of recombinant Ad-Mda7-tLyp1 and Ad-Mda7, the expression of recombinant proteins was analyzed by ELISA. Adenovectors were transduced (MOI=2-5) into Hep-G2, MCF7, MKN45, and normal skin fibroblast, then tumor-killing effect was measured by cytopathic effect (CPE) monitoring, MTT viability test, BAX gene expression analysis, and Caspase3/7 assay.

Results: ELISA assay revealed a sustained level of recombinant protein secretion following Adenovector transduction. In CPE microscopy, all cancer cell lines showed a significant reduction (≥50%) in their normal phenotype after receiving Ad-Mda7-tLyp1 and Ad-Mda7. The viability was significantly lower compared to the control, indicating an anti-proliferating effect. In parallel, the viability test showed that Ad-Mda7 and Ad-Mda7-tLyp1 have a significant killing effect (≥50%) on MCF-7, Hep-G2, and MKN45 compared to normal fibroblast (P≤0.05). BAX gene expression analysis showed that both Ad-Mda7-tLyp1 and Ad-Mda7 vectors induced >2-fold increase of apoptosis (P<0.05), particularly in MCF7. Similarly, caspase3/7 activity showed a significant increase (P<0.05) following Ad-Mda7, and Ad-Mda7-tLyp1 transduction into cancer cell lines, but not in normal fibroblasts.

Conclusion: The newly constructed Ad-Mda-tlyp1 showed a suitable tumor cell killing activity and enough specificity on studied cell lines.

Aim: In this study, we intend to evaluate the occurrence of small fiber neuropathy in patients with irritable bowel syndrome (IBS).

Background: Small fiber neuropathy (SFN) is a sensory neuropathy that results from the degeneration of small Aδ and unmyelinated C fibers. SFN manifests positive symptoms, such as tingling, burning, prickling, and aching, and negative symptoms, including numbness, tightness, and coldness. The SFN coexistence with other comorbidities (e.g., fibromyalgia, inflammatory bowel disease, celiac disease) has been reported in previous studies.

Patients and Methods: We conducted a cross-sectional study to assess the coexistence of SFN and IBS. Forty-two IBS patients and forty-three healthy individuals were asked to fill out the Michigan Neuropathy Screening Instrument (MNSI) questionnaire. Results greater than three (>3) were considered positive. Participants with positive MNSI questionnaire results were examined for any neuropathy signs according to the Utah Early Neuropathy Scale (UENS) examination. The participants with positive results for the questionnaire and examination were checked for the sural and the superficial peroneal nerve conduction study (NCS). Normal NCS represented intact large fibers and the diagnosis of SFN.

Results: Ten participants, 7 (16.7 %) in the IBS group and 3 (6.9 %) in the healthy group, had positive results for the questionnaire. Four participants were positive for the examination, with normal NCS, and they were classified as SFN positive participants. All four SFN diagnoses were from the IBS group. No one in the healthy group was diagnosed with SFN. We could find a significant statistical difference (p < 0.05) between the IBS group and the healthy group in terms of the prevalence of SFN diagnosis.

Conclusion: The correlation between SFN and IBS suggests the possibility of a generalized neuropathy syndrome characterized by widespread neuronal impairment. Thus, any peripheral neuropathy symptom in IBS patients (and potentially other chronic pain disorders) should be evaluated for SFN since timely diagnosis and proper treatment result in a better quality of life for the patients.

Aim: This study is done to investigate the hypolipidemic and hepatoprotective effects of corn silk extract in nicotine-administered male mice.

Background: Nicotine can induce pathophysiological effects in the liver tissue through oxidative stress and damage cells. Corn silk can improve liver function with its antioxidant effects.

Methods: In this experimental study, 30 male NMRI mice (25-30 gr) were divided into 5 groups: controls, sham, nicotine 2.5 mg/kg, nicotine+aqueous extract of corn silk 400 mg/kg, and nicotine+methanolic extract of corn silk 400 mg/kg for 1 month. One day after the last nicotine and extracts consumption, the serum samples were performed for biochemical measurement, and the supernatant of the homogenized liver was administered for antioxidant variables assessment.

Results: There was no significant difference in the body weight of different groups. Liver weight and GSH decreased in the nicotine group compared to the control group (P<0.05). Triglycerides, total cholesterol, HDL-C, LDL-C, liver enzymes, and MDA increased in the nicotine group compared to the control group (P<0.05). Also, the expansion of sinusoids, the presence of inflammatory cells, and necrosis of liver cells were observed in the nicotine group compared to the control group. Using aqueous and methanolic extracts of corn silk in mice receiving nicotine led to the improvement of the mentioned variables (P<0.05).

Conclusion: The results of this study showed that the use of nicotine can lead to the induction of hepatotoxicity. The use of aqueous and methanolic extracts of corn silk improved them through its antioxidant activity.

Aim: Due to the capabilities of the mobile application in the self-care of patients, the present study was conducted to design and evaluate a mobile-based self-care application for patients with liver cirrhosis.

Background: Liver cirrhosis is a progressive and chronic disease that, if left untreated, leads to liver cancer and, finally, the death of the patient.

Methods: This study was conducted in six phases, including determining and confirming the validity of the minimum data set and capabilities for the application, designing a conceptual and logical model and determining the technical capabilities, designing the application, evaluating the prototype usability in a laboratory environment by technical experts, evaluation of the application usability in a real environment by 30 patients with QUIS (Questionnaire of User Interface Satisfaction) questionnaire.

Results: The designed application has capabilities such as calculating the patient's MELD score (Model for End-Stage Liver Disease), medication reminder, location in emergency, and conversation with the physician. The results showed that the patients evaluated the application with a score of 7.94 (out of 9 points) at a good level. 

Conclusion: The self-care application can help patients with liver cirrhosis and their families access the necessary information related to the special care of the patient at any time and place; it also helps better manage the patient's life, improve the quality of life, and monitor the patient. These applications can effectively manage chronic diseases by reducing the burden of referrals and costs.

Aim: The current study aimed to introduce the key proteins involved in liver ischemia/reperfusion (I/R) injury through protein-protein interaction (PPI) analysis.

Background: Liver transplantation (LT) is a well-known treatment for liver diseases that threaten patients with mortality. LT is a complex operation, and several risks, including liver I/R injury, affect its success. Improving LT requires detection of its molecular mechanism. Experiments have revealed that high throughput methods such as proteomics in combination with bioinformatics are useful tools for analyzing the molecular mechanism of disease.

Methods: The differentially expressed proteins (DEPs) involved in liver I/R injury were extracted from the literature. The queried DEPs plus the first 100 neighbors were included in a network through STRING database using Cytoscape software. Degree, betweenness centrality, closeness centrality, and stress were considered to determine the central nodes. The queried DEPs were assessed by action map analysis using the CluePedia application of Cytoscape software. The key proteins were identified by comparing network analysis and action map evaluation results.

Results: Six proteins, namely ALB, INS, GAPDH, CAT, IL6, and TNF, among the added first neighbors were determined as the central first neighbors. MPO, CRP, MMP9, and HMOX1 were selected as central DEPs among the queried proteins. Action map analysis confirmed the PPI findings. The final evaluation revealed that MMP9 in combination with CRP and HMOX1 plays a critical role in liver I/R injury.

Conclusion: The significant role of MMP9 in liver I/R injury was detected in this study. Two central proteins (CRP and HMOX1) were shown to have a regulatory effect on MMP9; CRP activated MMP9, while HMXO1 downregulated it.

Hepatocellular carcinoma (HCC) typically presents with a primary hepatic mass. Nevertheless, on rare occasions, the initial presentation can be exclusively related to extrahepatic metastases and the most common sites of metastases are the lungs, lymph nodes, bones, and adrenal glands. While, bone metastases are generally accompanied by multiple metastatic spreads elsewhere in the body   or previously diagnosed HCC, cases of solitary bone metastases with no liver lesion at imaging have been reported. Indeed, two rare entities of HCC have been reported in the literature which are the ectopic hepatocellular carcinoma and the infiltrative type of hepatocellular carcinoma with a very challenging radiologic diagnosis and poor prognosis. In this article, we present a case of extrahepatic costal metastases of hepatocellular carcinoma, which was diagnosed through a bone biopsy, with no focal lesion on liver imaging including ultrasound, multiphase MRI, and CT scan except for the presence of a portal vein thrombosis. It is important to consider the possibility of HCC metastases when evaluating rapidly growing extrahepatic lesions in patients with chronic liver disease and to consider the tumor characteristics and imaging findings as well as limitations to make accurate and timely diagnosis leading to improved patient management. Our patient had probably an infiltrating HCC because of two prominent factors: the presence of portal vein thrombosis and a markedly elevated alpha-fetoprotein (AFP). A liver biopsy was crucial in order to confirm the diagnosis but unfortunately it could not be performed because of the unexpected death of the patient due to hemorrhagic shock. It is also worth noting in this case, that the elevated level of AFP raised the suspicion on an underlying HCC and contributed to more elaborate diagnostic tests.

Meckel diverticulum is the most common congenital anomaly of the gastrointestinal tract which is located in the small bowel within 2 feet of the ileocecal valve. However, an inverted Meckel's diverticulum is a rare disorder that seems to be caused by abnormal peristalsis in that region. This article demonstrates signs, symptoms and possible clinical presentations using CARE guideline in a case of inverted Meckel's diverticulum and review other possible features lastly, definitive treatment, results and case follow-up was shown to refresh and raise surgeons' awareness of this rare disorder.