A novel approach to improving colonoscopy learning efficiency through a colonoscope roaming system: randomized controlled trial.

Background: Colonoscopy is indispensable in the diagnosis and treatment of lower digestive tract (LDT) diseases. Skilled colonoscopists are in great demand, but it takes considerable time for beginners to become experts. In addition, patients may refuse to permit primary learners to practise colonoscopy on them. Thus, improving the instructional programmes and models for primary learners is a key issue in endoscopy training. Convenience and a self-paced, learner-centred approach make e-learning an excellent instructional prospect. Therefore, we created the Colonoscope Roaming System (CRS) to assist in colonoscopy teaching procedures. We aimed to develop the e-learning software, test it with beginner colonoscopists and evaluate its effectiveness via subjective and objective methods. Methods: Through a randomized controlled trial, participants were randomly allocated to an e-learning group (EG) or a control group (CG) after a pretest evaluation. The CG learned through the traditional colonoscopy teaching mode, while the EG used CRS in addition to the traditional teaching mode. Subsequent to the training, the participants completed a posttest and colonoscopy examination. The EG also completed a satisfaction questionnaire. Results: Of the 84 participants, 81 (96%) finished the colonoscopy learning and evaluation modules of the CRS. No conspicuous differences in the pretest scores were found between the EG and CG (p > 0.05). Two months later, the posttest scores for the EG were higher than those of the CG (p < 0.001), and the EG had better performance on the colonoscopy examination (p < 0.01). Overall, 86.25% of questions raised in Q1-Q20 were satisfied with the CRS and considered it successful. Conclusions: The use of CRS may be an effective approach to educate beginner colonoscopists to attain skills.

Construction of a TTN Mutation-Based Prognostic Model for Evaluating Immune Microenvironment, Cancer Stemness, and Outcomes of Colorectal Cancer Patients.

Background: Colorectal cancer (CRC) is one of the commonest cancers worldwide. As conventional biomarkers cannot clearly define the heterogeneity of CRC, it is essential to establish novel prognostic models. Methods: For the training set, data pertaining to mutations, gene expression profiles, and clinical parameters were obtained from the Cancer Genome Atlas. Consensus clustering analysis was used to identify the CRC immune subtypes. CIBERSORT was used to analyze the immune heterogeneity across different CRC subgroups. Least absolute shrinkage and selection operator regression was used to identify the genes for constructing the immune feature-based prognostic model and to determine their coefficients. Result: A gene prognostic model was then constructed to predict patient outcomes; the model was then externally validated using data from the Gene Expression Omnibus. As a high-frequency somatic mutation, the titin (TTN) mutation has been identified as a risk factor for CRC. Our results demonstrated that TTN mutations have the potential to modulate the tumor microenvironment, converting it into the immunosuppressive type. In this study, we identified the immune subtypes of CRC. Based on the identified subtypes, 25 genes were selected for prognostic model construction; a prediction model was also constructed, and its prediction accuracy was tested using the validation dataset. The potential of the model in predicting immunotherapy responsiveness was then explored. Conclusion: TTN-mutant and TTN-wild-type CRC demonstrated different microenvironment features and prognosis. Our model provides a robust immune-related gene prognostic tool and a series of gene signatures for evaluating the immune features, cancer stemness, and prognosis of CRC.

A clinical report on high­dose cytarabine therapy for children with acute myeloid leukemia.

Despite improvement in the long-term survival rate following pediatric acute myeloid leukemia (AML), the rate remains low, even with optimal treatment. The present study reports the long-term outcome of a small patient group treated with a single drug, high-dose chemotherapy (HDCT) with cytarabine, including consolidation and maintenance therapy. RT-PCR was conducted to assess 43 fusion genes, and after treatment, all cases have been followed up for 20 years (June 2002-December 2020). With an 80% 5-year survival rate, the results of this study highlight the possibility that pediatric AML can be reasonably effectively treated with relatively simple chemotherapy when necessary. HDCT is clinically safe, effective and relatively inexpensive. We propose that in the context of limited resources, HDCT should be considered as an alternative therapy for pediatric AML.

From Hair to Colon: Hair Follicle-Derived MSCs Alleviate Pyroptosis in DSS-Induced Ulcerative Colitis by Releasing Exosomes in a Paracrine Manner.

Ulcerative colitis (UC) has attracted intense attention due to its high recurrence rate and the difficulty of treatment. Pyroptosis has been suggested to be crucial in the development of UC. Although mesenchymal stem cells (MSCs) are broadly used for UC therapy, they have rarely been studied in the context of UC pyroptosis. Hair follicle-derived MSCs (HFMSCs) are especially understudied with regard to UC and pyroptosis. In this study, we aimed to discover the effects and potential mechanisms of HFMSCs in UC. We administered HFMSCs to dextran sulfate sodium- (DSS-) treated mice and found that the HFMSCs significantly inhibited pyroptosis to alleviate DSS-induced UC. A transwell system and GW4869, an exosome inhibitor, were used to prove the paracrine mechanism of HFMSCs. HFMSC supernatant reduced pyroptosis-related protein expression and promoted cell viability, but these effects were attenuated by GW4869, suggesting a role for HFMSC-released exosomes (Exos) in pyroptosis. Next, Exos were extracted and administered in vitro and in vivo to explore their roles in pyroptosis and UC. In addition, the biodistribution of Exos in mice was tracked using an imaging system and immunofluorescence. The results suggested that Exos not only improved DSS-induced pyroptosis and UC but also were internalized into the injured colon. Furthermore, the therapeutic efficacy of Exos was dose dependent. Among the Exo treatments, administration of 400 µg of Exos per mouse twice a week exhibited the highest efficacy. The differentially expressed miRNAs (DEmiRNAs) between MSCs and MSC-released Exos suggested that Exos might inhibit pyroptosis through tumour necrosis factor-related apoptosis-inducing ligand (TRAIL) signalling and interferon- (IFN-) gamma pathways. Our study reveals that HFMSCs can alleviate pyroptosis in UC by releasing DEmiRNA-containing Exos in a paracrine manner. This finding may lead to new treatments for UC.

Network Pharmacology-Based Study on the Mechanism of Gegen Qinlian Decoction against Colorectal Cancer.

PURPOSE: Gegen Qinlian decoction (GQD) has been used to treat gastrointestinal diseases, such as diarrhea and ulcerative colitis (UC). A recent study demonstrated that GQD enhanced the effect of PD-1 blockade in colorectal cancer (CRC). This study used network pharmacology analysis to investigate the mechanisms of GQD as a potential therapeutic approach against CRC. MATERIALS AND METHODS: Bioactive chemical ingredients (BCIs) of GQD were collected from the Traditional Chinese Medicine Systems Pharmacology (TCMSP) database. CRC-specific genes were obtained using the gene expression profile GSE110224 from the Gene Expression Omnibus (GEO) database. Target genes related to BCIs of GQD were then screened out. The GQD-CRC ingredient-target pharmacology network was constructed and visualized using Cytoscape software. A protein-protein interaction (PPI) network was subsequently constructed and analyzed with BisoGenet and CytoNCA plug-in in Cytoscape. Gene Ontology (GO) functional and the Kyoto Encyclopaedia of Genes and Genomes (KEGG) pathway enrichment analysis for target genes were then performed using the R package of clusterProfiler. RESULTS: One hundred and eighteen BCIs were determined to be effective on CRC, including quercetin, wogonin, and baicalein. Twenty corresponding target genes were screened out including PTGS2, CCNB1, and SPP1. Among these genes, CCNB1 and SPP1 were identified as crucial to the PPI network. A total of 212 GO terms and 6 KEGG pathways were enriched for target genes. Functional analysis indicated that these targets were closely related to pathophysiological processes and pathways such as biosynthetic and metabolic processes of prostaglandins and prostanoids, cytokine and chemokine activities, and the IL-17, TNF, Toll-like receptor, and nuclear factor-kappa B (NF-κB) signaling pathways. CONCLUSION: The study elucidated the "multiingredient, multitarget, and multipathway" mechanisms of GQD against CRC from a systemic perspective, indicating GQD to be a candidate therapy for CRC treatment.

Risks of Postendoscopic Retrograde Cholangiopancreatography Pancreatitis and Hyperamylasemia After Endoscopic Papillary Balloon Dilation: A Retrospective Analysis.

It is currently unclear whether endoscopic papillary balloon dilation (EPBD) is associated with increased severe postendoscopic retrograde cholangiopancreatography pancreatitis (PEP)-related morbidity owing to conflicting reports. This study aimed to investigate whether EPBD increases the risk of PEP and hyperamylasemia. Clinical data of patients with choledocholithiasis, treated at the Second Affiliated Hospital of Harbin Medical University from January 2015 to December 2016 were analyzed. Patients were divided into the EPBD group and endoscopic sphincterotomy (EST)+EPBD group, and their characteristics and PEP and hyperamylasemia incidences were compared. Incidences related to dilated balloon diameter were also analyzed. There were no significant differences in patient characteristics and the incidences of PEP (2.6% vs. 0%; P=0.257) and hyperamylasemia (4.4% vs. 5.6%; P=0.954) between the 2 groups. Results were similar even with different balloon dilatations. EPBD without endoscopic sphincterotomy did not increase the risk of PEP and hyperamylasemia. It is a safe option for choledocholithiasis patients.

Bortezomib protects against dextran sulfate sodium­induced ulcerative colitis in mice.

Bortezomib, a first­in­class proteasome inhibitor, is a standard method of treatment in multiple myeloma. In the present study, the therapeutic effect of bortezomib was evaluated in an ulcerative colitis model induced by dextran sulfate sodium (DSS) in mice, and the mechanism of action was also investigated. Mice were administered with 3% DSS drinking water for 7 consecutive days and then they were intraperitoneally treated with bortezomib (0.2, 0.6 or 1 mg/kg) for 1, 3 or 7 days. Mice in the control group and the DSS group were provided the same volume of PBS, respectively. Body weight, stool characteristics and hematochezia were observed. Serum levels of tumor necrosis factor­α (TNF­α), C­reactive protein (CRP), albumin (ALB) and colonic activity of superoxide dismutase (SOD) were evaluated using specific kits. The expression of the transcription factor nuclear factor­κB (NF­κB) p65 gene and the DNA­binding activity of NF­κB protein were also evaluated. Administration of bortezomib attenuates colonic inflammation in mice. After 3 or 7 days of treatment, Disease Activity Index (DAI) as well as histological scores and NF­κB p65 protein expression were significantly reduced in mice treated with bortezomib at a dose of 0.6 or 1 mg/kg/day. Furthermore, it was also revealed that bortezomib was able to reduce serum levels of CRP and TNF­α caused by DSS and increase the level of ALB in serum and the activity of SOD in colonic tissues. These results demonstrated that bortezomib exerts a protective effect on DSS­induced colitis, and its underlying mechanisms are associated with the NF­κB gene inhibition that mitigates colon inflammatory responses in intestinal epithelial cells.

Upregulation of ID2 antagonizes arsenic trioxide-induced antitumor effects in cancer cells.

AIMS AND BACKGROUND: Arsenic trioxide (ATO) strongly induces apoptosis and differentiation in acute promyelocytic leukemia, and induces cell cycle arrest in most solid tumors. Although many signaling pathways are involved in its antitumor mechanism, a detailed investigation of the transforming growth factor beta-bone morphogenetic protein signaling pathway has not been performed. METHODS AND STUDY DESIGN: A microarray containing 113 genes associated with the pathway was used to screen important molecules that participate in the antitumor effects of ATO. The expression levels of the inhibitors of DNA binding-2 (ID2) in 4 different types of cancer cells were determined by quantitative reverse transcription PCR and Western blotting. Human esophageal squamous cell carcinoma cell line Eca109 and pancreatic carcinoma cell line BxPC3 cells were transfected with siRNAs targeting ID2 and scrambled control siRNA. Cell proliferation was evaluated by methyl thiazolyl tetrazolium assay. RESULTS: Eighteen upregulated and 12 downregulated genes were identified. After verification at the transcriptional and translational levels in 4 different cancer cells, ID2 was identified as an ATO antitumor-associated protein. In addition, specific silencing of ID2 could enhance ATO-induced cell proliferation inhibition in cancer cells. CONCLUSIONS: A combination of ATO and ID2-targeted agents may have considerable therapeutic benefits in cancers.

Chicken skin mucosa surrounding adult colorectal adenomas is a risk factor for carcinogenesis.

OBJECTIVES: Transformation of normal mucosa to colorectal adenoma could occur over a span of 5 to 20 years, whereas transformation of colorectal adenoma to colorectal cancer could take an additional 5 to 15 years. This study aims to investigate whether chicken skin mucosa (CSM) surrounding adult colorectal adenomas may be a risk factor for carcinogenesis. METHODS: Patients with colorectal mucosa, colorectal adenomas without CSM, or colorectal adenomas with CSM were enrolled in this study. Immunohistochemistry and enzyme-linked immunosorbent assays were used to determine the expression levels of proliferation markers (ki-67 and COX-2) and apoptosis factors (survivin and caspase-3) in tissues. RESULTS: The expression of ki-67 was significantly higher in colorectal adenomas with CSM compared with colorectal adenoma tissues (P < 0.01). Colorectal adenocarcinoma showed significantly higher levels of COX-2 protein compared with normal colorectal mucosa and colorectal adenoma tissues (P < 0.001). COX-2 expression was significantly higher in adenomas with CSM compared with normal colorectal mucosa (P < 0.001). Adenomas with CSM and adenocarcinomas exhibited significantly higher levels of survivin when compared with colorectal adenoma without CSM and normal tissues (P < 0.001). Although we found no significant difference in caspase-3 levels between adenocarcinomas and adenomas with CSM, caspase-3 expression was significantly lower in these tissues when compared with colorectal adenomas without CSM and normal mucosa (P < 0.001). CONCLUSIONS: The biological characteristics of colorectal adenomas with CSM were different from those of colorectal adenomas without CSM. Colorectal adenomas with CSM exhibited active cell proliferation and inhibition of apoptotic pathways, suggesting an increased risk of carcinogenesis in these adenomas.

[Effects of acute lymphoblastic leukemia children bone marrow mesenchymal stem cells on drug resistance of K562/A02 cell line].

The aim of study was to investigate the effect of acute lymphoblastic leukemia (ALL) children bone marrow mesenchymal stem cells (MSC) on resistance of K562/A02 cells and its mechanism. MSC obtained from bone marrow of AL children were cultured and identified. The co-culture of MSC and K562/A02 and the culture of K562/A02 cell suspension alone was performed, of which 2 kinds of cells were treated with same concentration of adriamycin (ADM), and the rate of apoptosis was detected by flow cytometry, bcl-2 and bax of K562/A02 were detected by RT-PCR, while mdr1 gene level was detected by FQ-PCR. The results indicated that the MSC separation and proliferation were viable and steady. The apoptosis rate of the K562/A02 cells co-cultured with MSC was 1.97 ± 0.11%, while apoptosis rate of the K562/A02 cells cultured alone was 8.38 ± 0.29%, there was significant difference (p < 0.05). As compared with the K562/A02 cells cultured alone, the bcl-2 gene expression in K562/A02 cells co-cultured with MSC obviously increased; ratio of bcl-2/bax was obviously enhanced. The mdr1 gene level in K562/A02 co-cultured with MSC was no statistical different from K562/A02 cultured alone (p > 0.05), which suggested that adhesion co-cultured with MSC did not induce mdr1 expression higher than the culture of suspension. It is concluded that the MSC of ALL children can escape the leukemia cells from proapoptotic effect of drugs, the resistance of K562/A02 to ADM may be involved in enhancement of bcl-2 gene expression of K562/A02 cells co-cultured with MSC, but not in relation to mdr1 gene in K562/A02 cells themselves.

[Change of CD4(+) CD25(+) regulatory T cells and NK Cells in peripheral blood of children with acute leukemia and its possible significance in tumor immunity].

This study was purposed to investigate the changes of CD4(+) CD25(+) regulatory T cells and NK cells in peripheral blood of acute leukemia children at different stages, the function of immune system and the possible roles of the CD4(+) CD25(+) regulatory T cells as well as NK cells in leukemia immunity. The number and proportion of CD4(+) CD25(+) regulatory T cells and NK cells were detected by flow cytometry in the peripheral blood of 53 acute leukemia children, including 25 patients in new diagnosis and 28 patients in continuous complete remission (CCR), and were compared with that of 20 normal children. The results indicated that the mean proportion of CD4(+) CD25(+) CD127(+) in CD4(+) T cells of peripheral blood in newly diagnosed patients, patients with CCR and normal children were (9.55 +/- 2.41)%, (8.54 +/- 2.51)% and (6.25 +/- 0.85)% respectively, the mean proportions of CD4(+)CD25(+)CD127(+) in newly diagnosed patients and patients with CCR were higher than that in normal children, the mean proportion of CD4(+)CD25(+)CD127(+) in newly diagnosed patients were higher than that in patients with CCR (p < 0.05). At the same time, the NK cell count in patients with acute leukaemia decreased as compared with normal control, while after achieving CCR, the NK cell count in patients were also less than that in normal control (4.11 +/- 3.87% and 10.41 +/- 7.20% vs 14.06 +/- 5.95%, p < 0.05). It is concluded that the application of CD4(+), CD25(+) and CD127(+) to detect regulatory T cells is a simple, reproductive and accurate method, and the CD4(+) CD25(+) CD127(+) T cells can better reflect the proportion of CD4(+)CD25(+) regulatory T cells. The increase of regulatory T cells and decrease of NK cells in pediatric patients with acute leukemia indicate that the function of NK cells may be depressed. Treg T cells play a role in occurrence and development of leukemia, and are involved in down-regulating NK cell function.

[Expression of human ermap gene in umbilical cord blood mononuclear cells during differentiation and development towards erythroid lineage].

The aim of study was to explore the potential of human erythroid membrane associated protein (ERMAP) gene in erythroid cell differentiation and development, mononuclear cells (MNCs) were isolated from umbilical cord blood and induced to erythroid cell differentiation by SCF, IL-3 and EPO. The cell morphology was observed by using optical microscopy, the positive rate of cells was counted by biphenylamine staining and the ratios of CD36+/CD235a-, CD36+/CD235a+, CD36-/CD235a+ cells were detected by flow cytometry, the change of human ermap gene expression level was analyzed by using fluorescent quantitative PCR (FQ-PCR). The results showed that the ermap gene expression level increased while MNCs were induced to erythroid lineage after treatment with SCF, IL-3 and EPO. It is concluded that the human ermap gene plays an important role in differentiation and development of erythroid cells.

[A study on the apoptosis of gastric carcinoma cells induced by arsenic trioxide combined with Ad-IkappaBalphaM].

OBJECTIVE: To study the activation of nuclear factor-kappaB (NF-kappaB) in gastric carcinoma SGC-7901 cells after treating with As(2)O(3) and the enhancement of the therapeutic effect of As(2)O(3) with recombinant adenovirus IkappaBalphaM. METHODS: Culture of gastric carcinoma SGC-7901 cells was carried out. The cells both uninfected and infected with Ad-IkappaBalpha but not treated with As(2)O(3) were used as control. Electrophoretic mobility shift assay (EMSA) and immunohistochemistry were used to detect the activation of NF-kappaB in the cells after treatment of As(2)O(3) and the combination with Ad-IkappaBalphaM. MTT, Hoechst staining and TUNEL were used to assay the change of apoptosis induced by As(2)O(3) after infection with Ad-IkappaBalphaM. RESULTS: The results of EMSA and immunohistochemical method showed that after the treatment of As(2)O(3) the cells showed high activity of NF-kappaB. Simultaneous infection with Ad-IkappaBalphaM can inhibit the activation of NF-kappaB; MTT method indicated that after the treatment of As(2)O(3) infected with Ad-IkappaBalphaM apoptosis rate of the cells (59.2 +/- 2.5)% was higher than that of the cells treated with As(2)O(3) and infected with Ad-IkappaBalpha but not treated with As(2)O(3) (47.5 +/- 2.3)% and these neither infected nor treated (40.0 +/- 1.2%), P < 0.01. The result of Hoechst staining method indicated that, in the group of cells treated with As(2)O(3) and infected with Ad-IkappaBalphaM, apoptosis rate is (27.7 +/- 2.6)%, which was higher than the that of the cells infected with Ad-IkappaBalpha (18.3 +/- 1.5)% but not treated with As(2)O(3) and these neither infected nor treated (11.0 +/- 1.7%), P < 0.05. Hoechst staining method was in accordance with TUNEL technique; it was shown that in the group of cells treated with As(2)O(3) and infected with Ad-IkappaBalphaM, apoptosis rate was (31.1 +/- 2.5)%, being still higher than that of cells infected with Ad-IkappaBalpha but not treated with As(2)O(3) (20.7 +/- 2.1)% and these neither infected nor treated (13.0 +/- 1.7)%, P < 0.01. Therefore, infection with Ad-IkappaBalphaM can significantly increase the apoptosis induced by As(2)O(3). CONCLUSIONS: Gastric carcinoma cells treated with As(2)O(3) show activity of NF-kappaB. It is indicated that the activity of NF-kappaB may be the mechanism of the antagonism of gastric carcinoma cells against the apoptosis induced by As(2)O(3). Infection with Ad-IkappaBalphaM can effectively inhibit the activation of NF-kappaB in gastric carcinoma cells and increase the cell apoptosis induced by As(2)O(3).

[Preliminary study on the safety and pharmacodynamic action of low dose L-asparaginase].

OBJECTIVE: To investigate the safety and therapeutic effect of low dose (1000 U/m(2)) L-asparaginase (L-Asp) in the treatment of children with acute lymphoblastic leukemia (ALL). METHODS: Six patients were treated with low dose L-Asp after previously suffered severe side effects from standard dose L-Asp (5000 - 10,000 U/m(2)). Twenty-eight blood samples were obtained randomly from 5 of them. Plasma asparagine concentration was detected by reverse phase-high performance liquid chromatography (RP-HPLC). RESULTS: All the patients treated with low dose L-Asp showed no any toxic symptoms. The plasma asparagine levels in the patients were all above 5 micromol/L except case 4 (4.91 micromol/L) before receiving L-Asp, and were all decreased below 0.5 micromol/L five days after receiving low dose L-Asp, except case 3 (3.70 micromol/L), the results being like that of receiving standard dose L-Asp. CONCLUSION: Low dose L-Asp has definite efficacy for childhood ALL, while avoids serious side effects from standard dose L-Asp.

[Research on the pharmacokinetics and pharmacodynamics of L-asparaginase during its treatment of childhood acute lymphoblastic leukemia].

OBJECTIVE: To investigate the changes in the activity of Escherichia coli asparaginase (L-asp) and the concentration of asparagines (ASN) in the plasma of the acute lymphoblastic leukemia (ALL) children receiving L-asp containing chemotherapeutic protocol to explore more reasonable usage of L-asp in the treatment of childhood ALL. METHODS: L-asp containing hemotherapy regimen of VDLP was used, in which L-asp (10,000 U/m(2)) was administered intravenously every other day for 10 doses in 15 children with ALL. A total of 340 peripheral blood samples were collected at scheduled time points during the therapy and plasma L-asp activity (by spectrophotometric assay) and asparagines concentration (by RP-HPLC) were measured. RESULTS: During the administration of L-asp, the plasma L-asp activity was increasing gradually peaked after eight doses and then decreased gradually, while the plasma concentration of asparagines maintained in complete or nearly complete depletion status. After the therapy courses finished, a plasma L-asp activity above 100 U/L with asparagines almost complete depletion status was lasting for about seven days. CONCLUSION: The current L-asp containing chemotherapeutic protocols in which L-asp was administered in a dose of 10 000/m(2) intravenously every other day, are efficient enough for the depletion of plasma ASN.