Bioinformatics Analysis and Experimental Verification of Prognostic and Biological Significance of Autophagy-Related Long Non-Coding RNAs in Gastric Carcinoma

Background:Long non-coding RNAs(lncRNAs)play a vital role in autophagy modulation and tumor progression.However,the key lncRNAs and their functions in gastric cancer(GC)remain largely unknown.Methods:A bioinformatic analysis of GC patients’gene expression profiling data from the Cancer Genome Atlas database was performed to identify autophagy-related lncRNAs that are associated with predictive risk.Through Cox regression and Lasso regression analyses,the autophagy-related lncRNAs that are associated with prognosis were identified,and a novel prognostic model for GC was established.The model was then used to evaluate the clinical features and predictive risk of individuals with GC.By using two datasets,GSE 62254(n=300)and GSE 15459(n=192),from Gene Expression Omnibus,its effectiveness was verified.Gene set enrichment analysis according to hallmark and Kyoto Encyclopedia of Genes and Genomes were used to determine the possible biological roles of these lncRNAs.Furthermore,the HOXD antisense growth-associated long non-coding RNA(HAGLR)mechanism in GC was discovered through in vitro and in vivo experiments.Results:Six lncRNAs associated with autophagy in GC were identified,and a new prognostic risk model based on these lncRNAs was established.The six-lncRNA signature was significantly associated with adverse clinicopathological features and found to be an independent GC prognostic factor.The model was proven to be effective and robust by GSE62254 and GSE15459.According to gene set enrichment analysis,the six lncRNAs appeared to be tightly linked to autophagy-related and cancer-related mechanisms.HAGLR was also found to promote tumor growth by enhancing autophagy signaling in GC.Conclusion:A novel prognostic model integrating HAGLR that can effectively evaluate and predict the prognostic risk of GC patients was established.The results indicated that HAGLR promotes gastric cancer progression by enhancing autophagy and is anticipated to be a potential new target for the treatment of gastric cancer.

Risk factor profiles for gastric cancer prediction with respect to Helicobacter pylori:A study of a tertiary care hospital in Pakistan

BACKGROUND Gastric cancer(GC)is the fourth leading cause of cancer-related deaths worldwide.Diagnosis relies on histopathology and the number of endoscopies is increasing.Helicobacter pylori(H.pylori)infection is a major risk factor.AIM To develop an in-silico GC prediction model to reduce the number of diagnostic surgical procedures.The meta-data of patients with gastroduodenal symptoms,risk factors associated with GC,and H.pylori infection status from Holy Family Hospital Rawalpindi,Pakistan,were used with machine learning.METHODS A cohort of 341 patients was divided into three groups[normal gastric mucosa(NGM),gastroduodenal diseases(GDD),and GC].Information associated with socioeconomic and demographic conditions and GC risk factors was collected using a questionnaire.H.pylori infection status was determined based on urea breath test.The association of these factors and histopathological grades was assessed statistically.K-Nearest Neighbors and Random Forest(RF)machine learning models were tested.RESULTS This study reported an overall frequency of 64.2%(219/341)of H.pylori infection among enrolled subjects.It was higher in GC(74.2%,23/31)as compared to NGM and GDD and higher in males(54.3%,119/219)as compared to females.More abdominal pain(72.4%,247/341)was observed than other clinical symptoms including vomiting,bloating,acid reflux and heartburn.The majority of the GC patients experienced symptoms of vomiting(91%,20/22)with abdominal pain(100%,22/22).The multinomial logistic regression model was statistically significant and correctly classified 80%of the GDD/GC cases.Age,income level,vomiting,bloating and medication had significant association with GDD and GC.A dynamic RF GC-predictive model was developed,which achieved>80%test accuracy.CONCLUSION GC risk factors were incorporated into a computer model to predict the likelihood of developing GC with high sensitivity and specificity.The model is dynamic and will be further improved and validated by including new data in future research studies.Its use may reduce unnecessary endoscopic procedures.It is freely available.

Real-world efficacy and safety of tofacitinib treatment in Asian patients with ulcerative colitis

BACKGROUND Real-world data on tofacitinib(TOF)covering a period of more than 1 year for a sufficient number of Asian patients with ulcerative colitis(UC)are scarce.AIM To investigate the long-term efficacy and safety of TOF treatment for UC,including clinical issues.METHODS We performed a retrospective single-center observational analysis of 111 UC patients administered TOF at Hyogo Medical University as a tertiary inflammatory bowel disease center.All consecutive UC patients who received TOF between May 2018 and February 2020 were enrolled.Patients were followed up until August 2020.The primary outcome was the clinical response rate at week 8.Secondary outcomes included clinical remission at week 8,cumulative persistence rate of TOF administration,colectomy-free survival,relapse after tapering of TOF and predictors of clinical response at week 8 and week 48.RESULTS The clinical response and remission rates were 66.3%and 50.5%at week 8,and 47.1%and 43.5%at week 48,respectively.The overall cumulative clinical remission rate was 61.7%at week 48 and history of anti-tumor necrosis factor-alpha(TNF-α)agents use had no influence(P=0.25).The cumulative TOF persistence rate at week 48 was significantly lower in patients without clinical remission than in those with remission at week 8(30.9%vs 88.1%;P<0.001).Baseline partial Mayo Score was significantly lower in responders vs non-responders at week 8(odds ratio:0.61,95%confidence interval:0.45-0.82,P=0.001).Relapse occurred in 45.7%of patients after TOF tapering,and 85.7%of patients responded within 4 wk after re-increase.All 6 patients with herpes zoster(HZ)developed the infection after achieving remission by TOF.CONCLUSION TOF was more effective in UC patients with mild activity at baseline and its efficacy was not affected by previous treatment with anti-TNF-αagents.Most relapsed patients responded again after re-increase of TOF and nearly half relapsed after tapering off TOF.Special attention is needed for tapering and HZ.

Meta analysis of the efficacy of western medicine combined with Qiliqiangxin capsule versus western medicine alone in the treatment of chronic heart failure

Objective: To compare the clinical efficacy of conventional Western medicine combined with Qiliqiangxin capsule and western medicine alone in the treatment of chronic heart failure, and to prove that Qiliqiangxin capsule combined treatment has more advantages, providing reference for clinical decision-making in the treatment of chronic heart failure. Methods: Randomized controlled trials (RCTs) of conventional Western medicine treatment and Western medicine combined with Qiliqiangxin capsule in the treatment of chronic heart failure were searched in databases such as PubMed, Embase, Webofscience, CNKI, WanFang, VIP, and CBM. The bias risk assessment was conducted using the RCT tool recommended by Cochrane, and then the meta-analysis was performed using RevMan5.4 and Stata17 software. Compare the efficacy evaluation of cardiac function, left ventricular ejection fraction (LVEF), left ventricular end diastolic diameter (LVEDD), cardiac stroke output (SV), 6-minute walking test (6MWT), and N-terminal proBNP in the conventional western medicine combined with Qiliqiangxin capsule group (hereinafter referred to as the treatment group) and the conventional western medicine group (hereinafter referred to as the control group). Results: A total of 20 RCTs meeting the criteria were included, including 2953 patients, including 1508 in the treatment group and 1445 in the control group. The results of meta-analysis showed that the treatment group had significantly better cardiac function evaluation, LVEF, LVEDD, SV, 6MWT, and NT-proBNP improvement than the control group. Its central functional efficacy evaluation (OR=2.09,95% CI: 1.71-2.55, P<0.001), LVEF (WMD=7.05,95% CI: 5.30-8.79, P<0.00001), LVEDD (WMD=6.73, 95% CI: 3.18-10.29, P=0.0002), SV (WMD=6.73, 95% CI: 3.18-10.29, P=0.0002), 6MWT (SMD=0.70,95% CI: 0.54-0.87, P<0.00001), NT-proBNP (SMD=-1.95,95% CI: -2.5 2 to 1.38 (P<0.0001), with statistically significant differences. Conclusion: Conventional western medicine combined with Qiliqiangxin capsule can significantly improve the clinical efficacy of heart failure, improve LVEF, LVEDD, SV, and NT-proBNP index, and improve exercise tolerance. It is worth using for reference in the treatment.

Clinical characteristics and prognosis of non-small cell lung cancer patients with liver metastasis:A population-based study

BACKGROUND The presence of liver metastasis(LM) is an independent prognostic factor for shorter survival in non-small cell lung cancer(NSCLC) patients.The median overall survival of patients with involvement of the liver is less than 5 mo.At present,identifying prognostic factors and constructing survival prediction nomogram for NSCLC patients with LM(NSCLC-LM) are highly desirable.AIM To build a forecasting model to predict the survival time of NSCLC-LM patients.METHODS Data on NSCLC-LM patients were collected from the Surveillance,Epidemiology,and End Results database between 2010 and 2018.Joinpoint analysis was used to estimate the incidence trend of NSCLC-LM.Kaplan-Meier curves were constructed to assess survival time.Cox regression was applied to select the independent prognostic predictors of cancer-specific survival(CSS).A nomogram was established and its prognostic performance was evaluated.RESULTS The age-adjusted incidence of NSCLC-LM increased from 22.7 per 1000000 in 2010to 25.2 in 2013,and then declined to 22.1 in 2018.According to the multivariable Cox regression analysis of the training set,age,marital status,sex,race,histological type,T stage,metastatic pattern,and whether the patient received chemotherapy or not were identified as independent prognostic factors for CSS(P < 0.05) and were further used to construct a nomogram.The C-indices of the training and validation sets were 0.726 and 0.722,respectively.The results of decision curve analyses(DCAs) and calibration curves showed that the nomogram was well-discriminated and had great clinical utility.CONCLUSION We designed a nomogram model and further constructed a novel risk classification system based on easily accessible clinical factors which demonstrated excellent performance to predict the individual CSS of NSCLC-LM patients.

Early serum albumin changes in patients with ulcerative colitis treated with tacrolimus will predict clinical outcome

BACKGROUND Oral tacrolimus is a therapeutic agent for moderate to severe steroid-dependent or resistant ulcerative colitis(UC),but remission induction is difficult,and it is necessary to treat the patient while considering the next treatment.AIM To examine serum albumin(Alb)level as a prognostic factor for the therapeutic effect of tacrolimus in clinical practice.METHODS Forty-seven patients with UC treated with tacrolimus at our institution were divided into remission and failure groups(colectomy or switch to biologics),and the biological data at the start of observation and at weeks 1 and 2 were retrospectively examined.Kaplan-Meier and multivariate analyses were performed using Alb as a prognostic factor in UC treatment.RESULTS During the three months observed,17(36.2%)patients failed treatment with tacrolimus.A comparison between the failure and remission groups showed a significant difference only in Alb in week 2,and in the week 2/week 0 Alb ratio,which showed the rate of change in Alb.The cut-off value of the week 2/week 0 Alb ratio that predicted failure was 1,and its area under the curve was 0.751(95%CI:0.604-0.898).In the Kaplan-Meier analysis,a week 2/week 0 Alb ratio≤1 had a significantly higher failure rate than that of>1;Cox proportional hazard regression analysis also showed that a week 2/week 0 Alb ratio≤1 was an independent prognostic factor for failure within 3 mo after the start of tacrolimus treatment.CONCLUSION A week 2/week 0 Alb ratio≤1 predicts failure within 3 mo of tacrolimus administration for UC.High failure risk exists with week 2 Alb values≤1 on admission.

Evaluating mucosal healing using colon capsule endoscopy predicts outcome in patients with ulcerative colitis in clinical remission

AIM To examine whether second generation of colon capsule endoscopy(CCE-2) is acceptable for assessing the severity of mucosal inflammation and evaluating mucosal healing using CCE-2 is able to predict outcome in ulcerative colitis(UC) patients, especially in clinical remission.METHODS A total of 30 consecutive UC patients in clinical remission were enrolled to undergo CCE-2. Clinical remission was defined as clinical activity index(CAI) ≤ 4 according to Rachmilewitz index. The rate of total colon observation and colon cleansing level were evaluated. Severity of mucosal inflammation in UC was assessed according to the Mayo endoscopic subscore(MES) and Ulcerative Colitis Endoscopic Index of Severity(UCEIS). Relapsefree survival was assessed. Acceptability of CCE-2 was assessed using a questionnaire survey.RESULTS The rate of total colon observation within its battery life was 93.3%. The proportion of "excellent" plus "good" cleansing level was 73.3%. The rate of mucosal healing(MES 0, 1) assessed by CCE-2 was 77.0%. The relapse-free survival rate was significantly higher in MES 0, 1 than in MES 2, 3(P = 0.0435), and in UCEIS 0-3 than in UCEIS 4-8(P = 0.0211), whereas there was no significant difference between CAI 0 and CAI 1-4 groups. A questionnaire survey revealed an overall acceptability of CCE.CONCLUSION CCE-2 is acceptable for assessing the severity of mucosal inflammation in UC patients, especially in clinical remission. Evaluating mucosal healing using CCE-2 was able to predict outcome.

Novel multiplex stool-based assay for the detection of early-stage colon cancer in a Chinese population

BACKGROUND Stool DNA(sDNA)methylation analysis is a promising,noninvasive approach for colorectal cancer screening;however,reliable biomarkers for detecting early-stage colon cancer(ECC)are lacking,particularly in the Chinese population.AIM To identify a novel stool-based assay that can improve the effectiveness of ECC screening.METHODS A blinded case-control study was performed using archived stool samples from 125 ECC patients,and 125 control subjects with normal colonoscopy.The cohort was randomly divided into training and test sets at a 1.5:1 ratio.Targeted bisulfite sequencing(TBSeq)was conducted on five pairs of preoperative and postoperative sDNA samples from ECC patients to identify DNA methylation biomarkers,which were validated using pyrosequencing.By logistic regression analysis,a multiplex stool-based assay was developed in the training set,and the detection performance was further assessed in the test set and combined set.Theχ^(2)test was used to investigate the association of detection sensitivity with clinico-pathological features.RESULTS Following TBSeq,three hypermethylated cytosine-guanine sites were selected as biomarkers,including paired box 8,Ras-association domain family 1 and secreted frizzled-related protein 2,which differed between the groups and were involved in important cancer pathways.An sDNA panel containing the three biomarkers was constructed with a logistic model.Receiver operating characteristic(ROC)analysis revealed that this panel was superior to the fecal immunochemical test(FIT)or serum carcinoembryonic antigen for the detection of ECC.We further found that the combination of the sDNA panel with FIT could improve the screening effectiveness.In the combined set,the sensitivity,specificity and area under the ROC curve for this multiplex assay were 80.0%,93.6%and 0.918,respectively,and the performance remained excellent in the subgroup analysis by tumor stage.In addition,the detection sensitivity did not differ with tumor site,tumor stage,histological differentiation,age or sex,but was significantly higher in T4 than in T1-3 stage tumors(P=0.041).CONCLUSION We identified a novel multiplex stool-based assay combining sDNA methylation biomarkers and FIT,which could detect ECC with high sensitivity and specificity throughout the colon,showing a promising application perspective.

Maintenance time of sedative effects after an intravenous infusion of diazepam: A guide for endoscopy using diazepam

AIM: To examine whether the sedative effects assessed by psychomotor tests would depend on the cytochrome P450 (CYP ) 2C19 genotypes after an infusion regimen of diazepam commonly used forgastrointestinal endoscopy in Japan. METHODS: Fifteen healthy Japanese volunteers consisting of three different CYP2C19 genotype groups underwent a critical ? icker fusion test, an eye movement analysis and a postural sway test as a test for physical sedative effects, and a visual analog scale (VAS) symptom assessment method as a test for mental sedative effects during the 336 h period after the intravenous infusion of diazepam (5 mg). RESULTS: The physical sedative effects assessed by the critical flicker test continued for 1 h (t values of 5 min, 30 min and 60 min later: 4.35, 5.00 and 3.19, respectively) and those by the moving radial area of a postural sway test continued for 3 h (t values of 5 h, 30 h, 60 min and 3 h later: -4.05, -3.42, -2.17 and -2.58, respectively), which changed significantly compared with the baseline level before infusion (P < 0.05). On the other hand, the mental sedative effects by the VAS method improved within 1 h. The CYP2C19 genotype-dependent differences in the postinfusion sedative effects were not observed in any of the four psychomotor function tests. CONCLUSION: With the psychomotor tests, the objective sedative effects of diazepam continued for 1 h to 3 h irrespective of CYP2C19 genotype status and the subjective sedative symptoms improved within 1 h. Up to 3 h of clinical care appears to be required after the infusion of diazepam, although patients feel subjectively improved.

Low-dose radiation on spinal cord might be a new therapy for intractable chronic cancer and non-cancer pain

Intractable chronic pain is a great challenge in clinic.Central sensitization based on the positive changes of dendritic spines is the main mechanism of intractable chronic pain.And low-dose radiation has been proved to regulate the changes of dendritic spines negatively.Hence,we make a hypothesis that low-dose radiation could relieve cancer and noncancer pain through negatively regulating the shape and reducing the number and density of dendritic spines in the spinal cord.This method is supposed to be a new therapy for intractable chronic pain by expanding indication to non-cancer pain,translocating radiation site from where the tumor exists to special segments of spinal cord and keeping radiation dose at a low level.This therapy would be reliable for relieving non-cancer pain and supply more choices for relieving cancer pain.

Influence of interleukin polymorphisms on development of gastric cancer and peptic ulcer

Pro-inflammatory cytokines are produced in the gastric mucosa by inflammatory cells activated by chronic Helicobacter pylori (H. pylori) infection. Polymorphisms of these cytokine genes are associated with individual differences in gastric mucosal cytokine mRNA level, which result in differences in gastric mucosal inflammation, acid inhibition and gastroduodenal disease risk in response to H. pylori infection. Although polymorphisms of interleukin (IL)-1B, IL-1RN and TNF-A have been reported to relate well with gastric cancer and peptic ulcer risk, those of IL-2, IL-4, IL-6 and IL-8 genes are unclear. In combined analyses using data from previous studies, we found that the risk of gastric non-cardia cancer development was significantly associated with IL-4-168 C allele (OR: 0.81, 95% CI: 0.69-1.00) and IL-4-590 T allele carrier status (0.61, 0.53-0.73), and IL-6-174 G/G genotype (2<Abstract>Pro-inflammatory cytokines are produced in the gastric mucosa by inflammatory cells activated by chronic Helicobacter pylori (H. pylori) infection. Polymorphisms of these cytokine genes are associated with individual differences in gastric mucosal cytokine mRNA level, which result in differences in gastric mucosal inflammation, acid inhibition and gastroduodenal disease risk in response to H. pylori infection. Although polymorphisms of interleukin (IL)-1B, IL-1RN and TNF-A have been reported to relate well with gastric cancer and peptic ulcer risk, those of IL-2, IL-4, IL-6 and IL-8 genes are unclear. In combined analyses using data from previous stud- ies, we found that the risk of gastric non-cardia cancer development was significantly associated with IL-4-168 C allele (OR: 0.81, 95% CI: 0.69-1.00) and IL-4-590 T allele carrier status (0.61, 0.53-0.73), and IL-6-174 G/G genotype (2.02, 1.31-3.10). In peptic ulcer development, IL-2-330 G and IL-4-590 T allele carriers had a significantly decreased risk (0.37, 0.27-0.50 and 0.58, 0.34-0.99, respectively). Moreover, IL-2, IL-4, IL-6 and IL-8 gene genotypes prevalence differs among popula- tions. The inflammatory cytokine gene polymorphisms (e.g. IL-4 -590 and IL-6 -572 for gastric cancer, and IL-4-590, IL-6-572 and IL-8-251 for peptic ulcer) have a more potent influence on development of gastroduo- denal diseases in Western than East Asian populations. These cytokine gene polymorphisms, as well as those of IL-1B, IL-1RN and TNF-A, may be used to identify groups at higher risk of gastric cancer and peptic ulcer, and those suitable for their prevention by H. pylori eradication therapy in Western populations..02, 1.31-3.10). In peptic ulcer development, IL-2-330 G and IL-4-590 T allele carriers had a significantly decreased risk (0.37, 0.27-0.50 and 0.58, 0.34-0.99, respectively). Moreover, IL-2, IL-4, IL-6 and IL-8 gene genotypes prevalence differs among populations. The inflammatory cytokine gene polymorphisms (e.g. IL-4 -590 and IL-6 -572 for gastric cancer, and IL-4-590, IL-6-572 and IL-8-251 for peptic ulcer) have a more potent influence on development of gastroduo-denal diseases in Western than East Asian populations. These cytokine gene polymorphisms, as well as those of IL-1B, IL-1RN and TNF-A, may be used to identify groups at higher risk of gastric cancer and peptic ulcer, and those suitable for their prevention by H. pylori eradication therapy in Western populations.

Efficacy of tailored Helicobacter pylori eradication therapy based on antibiotic susceptibility and CYP2C19 genotype

The cure rates of Helicobacter pylori(H.pylori)eradication therapy using a proton pump inhibitor(PPI)and antimicrobial agents such as amoxicillin,clarithromycin,and metronidazole are mainly influenced by bacterial susceptibility to antimicrobial agents and the magnitude of the inhibition of acid secretion.Annual cure rates have gradually decreased because of the increased prevalence of H.pylori strains resistant to antimicrobial agents,especially to clarithromycin.Alternative regimens have therefore been developed incorporating different antimicrobial agents.Further,standard PPI therapy(twice-daily dosing)often fails to induce a long-term increase in intragastric pH>4.0.Increasing the eradication rate requires more frequent and higher doses of PPIs.Therapeutic efficacy related to acid secretion is influenced by genetic factors such as variants of the genes encoding drug-metabolizing enzymes(e.g.,cytochrome P450 2C19,CYP2C19),drug transporters(e.g.,multidrug resistance protein-1;ABCB1),and inflammatory cytokines(e.g.,interleukin-1β).For example,quadruple daily administration of PPI therapy potently inhibits acid secretion within 24 h,irrespective of CYP2C19 genotype.Therefore,tailored H.pylori eradication regimens that address acid secretion and employ optimal antimicrobial agents based on results of antimicrobial agent-susceptibility testing may prove effective in attaining higher eradication rates.

Characteristics of non-erosive gastroesophageal reflux disease refractory to proton pump inhibitor therapy

AIM:To investigate whether potent acid inhibition is effective in non-erosive reflux disease (NERD) refractory to standard rabeprazole (RPZ) treatment. METHODS:We treated 10 Japanese patients with NERD resistant to standard dosages of RPZ:10 mg or 20 mg od,20 mg bid,or 10 mg qid for 14 d. All patients completed a frequency scale for symptoms of gastroesophageal reflux disease questionnaire frequency scale for the symptoms of GERD (FSSG); and underwent 24 h pH monitoring on day 14. RESULTS:With increased dosages and frequency of administration of RPZ,median intragastric pH significantly increased,and FSSG scores significantly decreased. With RPZ 10 mg qid,potent acid inhibition was attained throughout 24 h. However,five subjects were refractory to RPZ 10 mg qid,although the median intragastric pH in these subjects (6.6,range:6.2-7.1) was similar to that in the remaining five responsive subjects (6.5,range:5.3-7.3). With baseline RPZ 10 mg od,FSSG scores in responsive patients improved by > 30%,whereas there was no significant decrease in the resistant group. CONCLUSION:NERD patients whose FSSG score fails to decrease by > 30% after treatment with RPZ 10 mg od for 14 d are refractory to higher dosage.

Association between endotoxemia and histological features of nonalcoholic fatty liver disease

AIM To assess whether surrogate biomarkers of endotoxemia were correlated with the histological features ofnonalcoholic fatty liver disease(NAFLD).METHODS One hundred twenty-six NAFLD patients who had undergone percutaneous liver biopsy were enrolled. Serum lipopolysaccharide(LPS)-binding protein(LBP) and anti-endotoxin core immunoglobulin G(Endo Cab Ig G) antibody concentrations at the time of liver biopsy were measured using the enzyme-linked immunosorbent assays to examine for relationships between biomarker levels and histological scores. RESULTS Serum LBP concentration was significantly increased in nonalcoholic steatohepatitis(NASH) patients as compared with nonalcoholic fatty liver(NAFL) subjects and was correlated with steatosis(r = 0.38, P < 0.0001) and ballooning scores(r = 0.23, P = 0.01), but not with the severity of lobular inflammation or fibrosis. Multivariate linear regression analysis revealed that LBP was associated with steatosis score and circulating C-reactive protein, aspartate aminotransferase, and fibrinogen levels. Serum Endo Cab Ig G concentration was comparable between NASH and NAFL patients. No meaningful correlations were detected between Endo Cab Ig G and histological findings. CONCLUSION LBP/Endo Cab Ig G were not correlated with lobular inflammation or fibrosis. More accurate LPS biomarkers are required to stringently assess the contribution of endotoxemia to conventional NASH.

Analysis of the genetic interactions between Cyclin A1, Atm and p53 during spermatogenesis

Aim： To analyze the functional interactions of Cyclin with p53 and Atm in spermatogenesis and DNA double- strand break repair. Methods： Two lines of double knockout mice were generated. Spermatogenesis and double strand break repair mechanisms were analyzed in Cyclin A1 （Ccnal）; p53- and Ccnal; Atm-double knockout mice. Results： The block in spermatogenesis observed in Cyclin A1-/- （Ccnal-/-） testes at the mid-diplotene stage is associated with polynucleated giant cells. We found that Ccnal-deficient testes and especially the giant cells accumulate unrepaired DNA double-strand breaks, as detected by immunohistochemistry for phosphorylated H2AX. In addition, the giant cells escape from apoptosis. The development of giant cells occurred in meiotic prophase I, because testes lacking ATM, which are known to develop spermatogenic arrest earlier than prophase I, do not develop giant cells in the absence of cyclin A1. Cyclin A1 interacted with p53 and phosphorylated p53 in complex with CDK2. Interestingly, p53-deficiency significantly increased the number of giant cells in Ccnal-deficient testes. Gene expression analyses of a panel of DNA repair genes in the mutant testes revealed that none of the genes examined were consistently misregulated in the absence of cyclin A1. Conclusion： Ccnal-deficiency in spermatogenesis is associated with defects in DNA double-strand break repair, which is enhanced by loss of p53.

A case of small bowel adenocarcinoma in a patient with Crohn’s disease detected by PET/CT and double-balloon enteroscopy

Small bowel adenocarcinoma (SBA) in patients with Crohn’s disease (CD) is quite rare, difficult to diagnose without surgery, and has a poor prognosis. Here, we report a 48-year-old man with SBA and a 21-year history of CD who was diagnosed by a combination of positron emission tomography/computed tomography (PET/CT) and double-balloon enteroscopy (DBE). Since the age of 27 years, the patient had been treated for ileal CD and was referred to our hospital with persistent melena. Multiple hepatic tumors were found by CT. PET/CT detected an accumulation spot in the small bowel. DBE revealed an ulcerative tumor in the ileum about 100 cm from the ileocecal valve. An endoscopic forceps biopsy specimen showed poorly differentiated adenocarcinoma. There were some longitudinal ulcer scars near the tumor, and the chronic inflammation inthe small bowel appeared to be associated with the cancer development. Previous reports suggest the risk of SBA in patients with CD is higher than in the overall population. Since early diagnosis is extremely difficult in these cases, novel techniques, such as PET/CT and DBE, may be expected to help in making a preoperative diagnosis of the development of SBA in CD.

Inverted Meckel's diverticulum preoperatively diagnosed using double-balloon enteroscopy

An inverted Meckel's diverticulum is a rare gastrointestinal congenital anomaly that is difficult to diagnose prior to surgery and presents with anemia, abdominal pain, or intussusception. Here, we report the case of 57-year-old men with an inverted Meckel's diverticulum, who was preoperatively diagnosed using doubleballoon enteroscopy. He had repeatedly experienced epigastric pain for 2 mo. Ultrasonography and computed tomography showed intestinal wall thickening in the pelvis. Double-balloon enteroscopy via the anal route was performed for further examination, which demonstrated an approximately 8-cm, sausage-shaped, submucosal tumor located approximately 80 cm proximal to the ileocecal valve. A small depressed erosion was observed at the tip of this lesion. Forceps biopsy revealed heterotopic gastric mucosa. Thus, the patient was diagnosed with an inverted Meckel's diverticulum, and single-incision laparoscopic surgery was performed. This case suggests that an inverted Meckel's diverticulum should be considered as a differential diagnosis for a submucosal tumor in the ileum. Balloon-assisted enteroscopy with forceps biopsy facilitate a precise diagnosis of this condition.

Post-cholecystectomy symptoms were caused by persistence of a functional gastrointestinal disorder

AIM:To classify gallstone disease as a basis for assessment of post-cholecystectomy symptoms.METHODS:One hundred and fifty three patients with a clinical and ultrasonographic diagnosis of gallstones filled out a structured questionnaire on abdominal pain symptoms and functional gastrointestinal disorder (FGID) before and at six months after cholecystectomy.Symptom frequency groups (SFG) were categorized according to frequency of pain attacks.According to certain pain characteristics in gallstone patients,a gallstone symptom score was accorded on a scale from one to ten.A visual analogue scale was used to quantify pain.Operative specimens were examined for size and magnitude of stone contents as well as presence of bacteria.Follow-up took place after six months with either a consultation or via a mailed questionnaire.Results were compared with those obtained pre-operatively to describe and analyze symptomatic outcome.RESULTS:SFG groups were categorized as severe (24.2%),moderate (38.6%) and mild (22.2%) attack frequency,and a chronic pain condition (15%).Pain was cured or improved in about 90% of patients and two-thirds of patients obtained complete symptom relief.Patients with the most frequent pain episodes were less likely to obtain symptom relief.FGID was present in 88% of patients pre-operatively and in 57% postoperatively (P=0.244).Those that became asymptomatic or improved with regard to pain also had most relief from FGID (P=0.001).No pre-operative FGID meant almost complete cure.CONCLUSION:Only one third of patients with FGID experienced postoperative relief,indicating that FGID was a dominant cause of post-cholecystectomy symptoms.

Tuberous sclerosis patient with neuroendocrine carcinoma of the esophagogastric junction:A case report

BACKGROUND Tuberous sclerosis complex(TSC)is a rare inherited disease with non-cancerous tumor growths in the skin,brain,kidneys,heart,and lungs.The co-occurrence of neuroendocrine neoplasm(NEN)with TSC is even rarer.There have been few reports on the relationship between TSC and neuroendocrine tumors(NETs),and fewer on the relationship between TSC and neuroendocrine carcinoma(NEC),a subtype of NEN.This is the first reported case of NEC occurring at the esophagogastric junction in a patient with TSC.CASE SUMMARY A 46-year-old woman visiting our hospital for the treatment of TSC was admitted to the emergency department with tarry stools and dizziness.Computed tomography scans revealed thickness of the gastric cardia,multiple metastatic lesions of the liver,and enlarged lymph nodes near the lesser curvature of the stomach.Esophagogastroduodenoscopy revealed a type 3 tumor located from the esophagogastric junction to the fundus,and the pathological diagnosis by biopsy was NEC.The patient was treated with seven courses of cisplatin+irinotecan,followed by eight courses of ramucirumab+nab-paclitaxel,one course of nivolumab,and two courses of S-1+oxaliplatin.Twenty-three months after the first treatment,the patient died because of disease progression and deterioration of the general condition.CONCLUSION This case of NEC occurring in a patient with TSC indicates a difference in the occurrence of NETs and NECs.

Neuroprotective effects of naturally sourced bioactive polysaccharides: an update

Polysaccharides are macromolecular complexes that have various biological activities.In vivo and in vitro studies have shown that polysaccharides play neuroprotective roles through multiple mechanisms;consequently,they have potential in the prevention and treatment of neurodegenerative diseases.This paper summarizes related research published during 2015-2020 and reviews advances in the understanding of the neuroprotective effects of bioactive polysaccharides.This review focuses on 15 bioactive polysaccharides from plants and fungi that have neuroprotective properties against oxidative stress,apoptosis,neuroinflammation,and excitatory amino acid toxicity mainly through the regulation of nuclear factor kappa-B,phosphatidylinositol-3-kinase/protein kinase B,mitogen-activated protein kinase,nuclear factor-E2-related factor 2/hemeoxygenase-1,c-jun N-terminal kinase,protein kinase B-mammalian target of rapamycin,and reactive oxygen species-nucleotide-binding oligomerization domain,leucine-rich repeat and pyrin domain-containing 3 signaling pathways.Natural bioactive polysaccharides have potential in the prevention and treatment of neurodegenerative diseases because of their advantageous characteristics,including multi-targeting,low toxicity,and synergistic effects.However,most of the recent related research has focused on cell and animal models.Future randomized clinical trials involving large sample sizes are needed to validate the therapeutic benefits of these neuroprotective polysaccharides in patients having neurodegenerative diseases.