Endoscopic resection vs. endoscopic resection plus chemoradiation for T1 stage colorectal cancer: a real-world retrospective cohort study.

Background: Early-stage colorectal cancer (CRC) patients treated with either endoscopic resection (ER) alone or combined ER with chemoradiotherapy (CRT) have unknown survival rates. A national descriptive epidemiological study was conducted to compare the long-term survival of patients with T1 stage CRC with or without the two different treatment options. Methods: Our study identified the records of patients with T1-stage CRC between 2010 and 2018 by searching the Surveillance, Epidemiology, and End Results (SEER) database. Long-term survival was compared using Kaplan-Meier methods and Cox proportional hazard models based on patient demographic and cancer parameters. Results: After propensity score matching (PSM), 825 T1-stage CRC patients were finally enrolled in this study, with 718 patients treated with ER and 107 patients treated with ER + CRT. The overall survival (OS) and cancer specific survival (CSS) rates were similar between the two treatment options (OS: P=0.47; CSS: P=0.28). According to subgroup analysis, older patients and patients with rectal tumor locations exhibited significantly higher OS and CSS rates in the ER + CRT group than in the ER group (OS: P<0.0001; CSS: P<0.0001). Conclusions: The findings from the SEER database showed that OS and CSS rates were similar between the ER and ER + CRT treated groups. Older patients and patients with rectal cancer benefited the most from ER + CRT treatment.

Comparison of statins with steroids and botulinum toxin A in the prevention of benign strictures after esophageal endoscopic submucosal dissection: a retrospective cohort study.

BACKGROUND: Preventing benign strictures following esophageal endoscopic submucosal dissection (ESD) remains difficult, and finding a safe, effective, and simple management method is vital. We previously reported that rosuvastatin significantly reduced the incidence and severity of strictures in a rabbit model of esophageal stricture. Accordingly, in this study, we compared the effects of statins, steroids, and botulinum toxin A (BTX-A) on stricture prevention after ESD involving more than three-fourths of the luminal circumference. METHODS: Of the 1019 ESD cases treated between January 2015 and December 2020, 246 met the inclusion criteria, with 21 cases excluded due to loss to follow-up, tumor recurrence, death, or need for additional surgery, radiotherapy, and/or chemotherapy. Of the 225 included cases, 145 received no intervention, while the remaining 80 were treated: 16 with oral steroids, 20 with topical triamcinolone acetonide (TA) injection, 21 with topical BTX-A injection, and 23 with statins. RESULTS: The occurrence stricture rate in the statins group (17.4%, 4/23) was significantly lower than in the non-intervention (75.2%, 109/145, P = 0.000), oral steroids (56.3%, 9/16, P = 0.011) and TA injection (50%, 10/20, P = 0.023) groups, but comparable to in the BTX-A injection (38.1%, 8/21, P = 0.124) group. The dysphagia score was lower in the statin than non-intervention group (P = 0.000). Although there was no significant difference in the number of required dilations between groups, the maximum number of dilations in the statins group was only six. CONCLUSIONS: Statins may be a potential treatment to prevent esophageal strictures after extensive ESD; however, clinical trials should be conducted to validate this.

A Multimodal Multipath Artificial Intelligence System for Diagnosing Gastric Protruded Lesions on Endoscopy and Endoscopic Ultrasonography Images.

INTRODUCTION: The aim of this study was to develop a novel artificial intelligence (AI) system that can automatically detect and classify protruded gastric lesions and help address the challenges of diagnostic accuracy and inter-reader variability encountered in routine diagnostic workflow. METHODS: We analyzed data from 1,366 participants who underwent gastroscopy at Jiangsu Provincial People's Hospital and Yangzhou First People's Hospital between December 2010 and December 2020. These patients were diagnosed with submucosal tumors (SMTs) including gastric stromal tumors (GISTs), gastric leiomyomas (GILs), and gastric ectopic pancreas (GEP). We trained and validated a multimodal, multipath AI system (MMP-AI) using the data set. We assessed the diagnostic performance of the proposed AI system using the area under the receiver-operating characteristic curve (AUC) and compared its performance with that of endoscopists with more than 5 years of experience in endoscopic diagnosis. RESULTS: In the ternary classification task among subtypes of SMTs using modality images, MMP-AI achieved the highest AUCs of 0.896, 0.890, and 0.999 for classifying GIST, GIL, and GEP, respectively. The performance of the model was verified using both external and internal longitudinal data sets. Compared with endoscopists, MMP-AI achieved higher recognition accuracy for SMTs. DISCUSSION: We developed a system called MMP-AI to identify protruding benign gastric lesions. This system can be used not only for white-light endoscope image recognition but also for endoscopic ultrasonography image analysis.

"M1/M2" Muscularis Macrophages Are Associated with Reduction of Interstitial Cells of Cajal and Glial Cells in Achalasia.

BACKGROUND AND AIMS: Several studies showed muscularis macrophages (MMφ) are associated with GI motility disorders. The purpose of this study was to preliminary explore the association between MMφ and achalasia. METHODS: Tissue samples of the lower esophageal sphincter (LES) high-pressure zone were obtained from 27 achalasia patients and 10 controls. Immunohistochemistry for MMφ, interstitial cells of Cajal (ICC), neuronal nitric oxide synthase (nNOS), and glial cells were conducted. Histological characteristics were compared between groups, and correlation analysis was performed. RESULTS: Fewer ICC was found in achalasia compared with controls (P = 0.018), and the level of M1 macrophages was higher than that in controls no matter in terms of the number or the proportion of M1(P = 0.026 for M1 and 0.037 for M1/MMφ). Statistical differences were found between two groups in terms of proportion of M2 and ratio of M1 to M2 (P = 0.048 for M2/ MMφ and < 0.001 for M1/M2). For the correlation analysis, significant correlations were detected between levels of nNOS, ICC, and glial cells in patients with achalasia (P = 0.026 for nNOS and ICC, 0.001 for nNOS and glial cells, 0.019 for ICC and glial cells). There were significant correlations between M2/MMφ and levels of ICC (P = 0.019), glial cells (P = 0.004), and nNOS (P = 0.135). CONCLUSION: Patients with achalasia had a higher level of M1/M2 ratio in LES and significant correlations were found between M2/MMφ and numbers of ICC and glial cells, which suggested that MMφ were probably associated with occurrence and development of achalasia.

The Long-Term Outcomes of Surgery Versus Endoscopic Treatment in Patients With Siewert Type II T1M0N0 Adenocarcinoma of the Esophagogastric Junction.

OBJECTIVE: We compared the long-term prognosis of surgery and endoscopic treatment (ET) in patients diagnosed with Siewert Type II pT1N0M0 adenocarcinoma of the esophagogastric junction (AEG). METHODS: Using the Surveillance, Epidemiology, and End Results (SEER) database, we performed a real-world retrospective cohort study and enrolled patients with Siewert Type II pT1N0M0 AEG who underwent surgery or endoscopic treatment (ET) from 2010 to 2018. Matched cohorts were generated using propensity score matching Competing-risk analysis was applied. The cumulative incidence function was used to calculate cancer-specific death and other causes of death (OCD) at different time points. Univariate and multivariate analyses were performed to identify significant prognostic factors by using the subdistribution hazard ratio. RESULTS: We enrolled 725 patients: 462 underwent surgery and 263 received ET. The 5 year cumulative CSD incidence significantly differed between surgery and ET cohorts (16.87% vs 11.08%, P = .01). Following PSM, 2 balanced groups (n = 219 patients each) were analyzed. No significant difference in the 5 year cumulative incidences of CSD was noted between cohorts (17.61% vs. 12.16%, P = .14). In multivariable analysis, the CSD incidence was high among patients with aged ≥65 (SHR 2.29, 95%CI 0.99-5.33, P = .05) and T1b-stage (SHR 1.92, 95%CI 1.03-3.57, P = .04); treatment (surgery or ET) was not significantly associated with cancer survival (SHR 1.51, 95% CI 0.81-2.81, P = .20). CONCLUSION: Long-term survival did not significantly differ among patients with Siewert Type II pT1N0M0 AEG adenocarcinoma undergoing surgery or ET. ET may be considered in patients >65 years old or those with submucosal (T1b-stage) cancer of AEG.

Establishment of a model for predicting delayed post-polypectomy bleeding: A real-world retrospective study.

Background: Delayed post-polypectomy bleeding (DPPB) is the most common complication which occurs within 30 days after polypectomy, it has become rather common with the widespread of colorectal cancer screening. It is important to clarified predictors of DPPB and identify patients at high risk. Materials and methods: This was a real-world retrospective study based on medical records from The First Affiliated Hospital of Nanjing Medical University. Cases of patients who underwent colonoscopic polypectomy between January 2016 and December 2020 were reviewed to identify risk factors of DPPB. We use the LASSO-Logistic regression analysis model to identify independent predictors and create a predictive model. The model finally got visualized by developing a nomogram. Results: Colonoscopic polypectomy was done on 16,925 patients in our study. DPPB occurred in 125 (0.74%) of these instances. In multivariate analysis, age, sex, hypertension, polyp location, polyp size, and operative modality were found to be independent risk factors and were integrated for the construction of a nomogram. The model's C-index is 0.801 (95%CI: 0.761-0.846). We also found polyps located at the right semicolon and polyp ≥ 1 cm associated with active bleeding under the therapeutic colonoscopy. Conclusion: Young age, male, hypertension, polyp ≥ 1 cm, proximal colon location and operative modality were finally identified as significant predictors of DPPB. We developed and validated a nomogram which performs well in predicting the incidence of DPPB, the model we established can be used as a valuable screening tool to identify patients who are at high risk of bleeding.

Risk factors for the failure of endoscopic resection of gastric submucosal tumors: a long-term retrospective case-control study.

OBJECTIVE: Endoscopic resection (ER) is an effective treatment method for gastric submucosal tumors (G-SMTs), but endoscopic resection failure requires emergency surgery. The purpose of this study was to assess potential risk factors for endoscopic resection failure. METHODS: A total of 1041 patients with G-SMT undergoing endoscopic resection were enrolled. Twenty-five patients in whom endoscopic resection failed, requiring a transition to surgery midway through the operation, were included in the failed group, and 1016 patients who received successful endoscopic resection were included in the successful endoscopic resection group. Baseline and lesion characteristics were recorded, and the differences in tumor characteristics and risk factors for resection failure of G-SMT were analyzed. Sensitivity analysis was performed to detect the stability of the indicator. RESULTS: Of the 1041cases included, there were 25 cases (2.4%) of failed endoscopic resection. Binary logistic analysis showed that the independent risk factors included tumors originating from deep muscularis propria(OR = 14.42, 95% CI 4.47-46.52), size > 3 cm (OR = 7.75, 95% CI 2.64-22.70), exophytic growth pattern (OR = 4.98, 95% CI 1.62-15.29), endoscopist with less experience (OR = 5.99, 95% CI 1.07-12.19), and irregular borders (OR = 4.13, 95% CI 1.40-12.19). The stable risk factors were tumors size, tumor origin and growth pattern according to sensitivity analysis. CONCLUSIONS: Tumors originating from the deep muscularis propria, tumor size > 3 cm, endoscopists with less experience, an exophytic growth pattern, and irregular boundaries were found to be independent risk factors for endoscopic resection failure. To reduce the risk of endoscopic resection failure, physicians should carefully evaluate G-SMT characteristics preoperative.

Treatment Options for T1 Stage Adenocarcinoma of Esophagogastric Junction: A Real-World Retrospective Cohort Study.

BACKGROUND: The number of patients diagnosed with T1 stage adenocarcinoma of esophagogastric junction (AEGJ) has been increasing. This study was conducted to investigate the effect of different treatment options (surgery, chemoradiation, and surgery+chemoradiation) on long-term survival in patients with T1-stage AEGJ. METHODS: We searched the Surveillance, Epidemiology, and End Results (SEER) database to identify the records of patients with T1-stage AEGJ between 2010 and 2018. Patient demographics and cancer parameters were compared among the three groups. The Kaplan-Meier method and Cox proportional hazard modeling were used to compare long-term survival. RESULTS: Data from 925 T1 stage AEGJ patients (surgery: n=516, surgery+chemoradiation: n=206, chemoradiation: n=203) were collected. We found that the OS and CSS rates of three treatment options had significant difference. Besides, positive nodal status also showed lower OS and CSS rat. Multivariate Cox regression analysis showed that surgery group has much lower risk of death compared with chemoradiation group and similar risk of death compared with surgery+chemoradiation group. Subgroup analysis suggested that in patients with N1-N3 status had higher OS and CSS rates in surgery+chemoradiation group. CONCLUSION: Using SEER data, we identified a significant survival advantage with the use of surgery compared to chemoradiation in patients with T1-stage AEGJ while the long-term survival of patients after surgery+chemoradiation group was not significantly different and low risk of death in positive nodal status.

Comparison of endoscopic injection of botulinum toxin and steroids immediately after endoscopic submucosal dissection to prevent esophageal stricture: a prospective cohort study.

Background: Widespread endoscopic submucosal dissection (ESD) in early esophageal cancer patients is closely associated with esophageal stricture, which dramatically reduces patients' quality of life and increases huge medical burdens. Endoscopic injection of steroid was proved as a protective method for post-ESD strictures. Other materials such as botulinum toxin type A (BTX-A) may be potential candidates. We conducted this prospective cohort study to compare the efficacy and feasibility of endoscopic injection of BTX-A and triamcinolone acetonide (TA) for the prevention of esophageal stricture. Methods: Seventy-eight patients with esophageal mucosal defects of more than two thirds of the circumference were successively enrolled and divided into 3 groups: BTX-A group (group A, n=26), TA group (group B, n=16) and control group (group C, n=36). Patients in group A were immediately injected with BTX-A after ESD, in group B were immediately injected with TA and in group C received ESD only. Endoscopy was performed when patients reported dysphagia symptoms and at 6 and 12 weeks post-ESD in patients without symptoms. Patients who experienced post-ESD esophageal strictures in all groups received bougie dilation. All patients were followed up for one year. Results: The proportion of patients developing stricture in BTX-A group was 30.00% (intention to treat analysis, 9/30) and 26.92% (per protocol analysis, 7/26), in TA group was 40.90% (intention to treat analysis, 9/22) and 43.75% (per protocol analysis, 7/16), and in control group was 84.21% (intention to treat analysis, 32/38) and 83.33% (per protocol analysis, 30/36) (p<0.001). When further comparing between each of the two groups, the incidence of esophageal stricture was lower in BTX-A group than that in control group (p<0.001), and lower in TA group than that in control group (p=0.004). Furthermore, in entire circumference mucosal defect subgroup, the esophageal stricture was significantly lower in BTX-A group than that in TA group (33.3% vs 100%, p=0.0454). Conclusions: Endoscopic injection of BTX-A and TA were effective in preventing post-ESD esophageal strictures and BTX-A injection was particularly effective in entire circumference mucosal defect patients. Multi-centered, randomized prospective study with larger sample size should be conducted. (Clinical trial registration number: ChiCTR2100042970, registered 1 February 2021, retrospectively registered, http://www.chictr.org.cn/listbycreater.aspx).

Predictive Value of Routine Blood Test in patients with Early Esophageal Cancer: A Matched Case-Control Study.

Aims: The present study was to evaluate the diagnostic value of routine blood test as potential inflammatory markers in early esophageal cancer (EEC) patients. Methods: A matched case-control study was conducted by recruiting 314 patients who were pathologically diagnosed with EEC and then underwent Endoscopic Submucosal Dissection (ESD) from July 2015 to July 2019 in First Affiliated Hospital of Nanjing Medical University. Each EEC patient was matched against one healthy control on the criteria of gender, and age (±2 years). Additionally, a total of 40 subjects (20 cases and 20 controls) were also included in the validation set. Statistical analysis of selected hematological parameters was performed between the two groups. The correlation between preoperative blood indexes and clinicopathological characteristics after ESD in EEC patients were further assessed. Results: Mono-factor analysis showed that the index of monocyte (p<0.001), MCV (p=0.018), MCH (p=0.01), MPV (p=0.022), PT (p=0.003), PT-INR (p=0.003), PDW (p<0.001) and MLR (p<0.001) were statistically significant in EEC patients when compared with those in healthy controls. Multivariate logistic regression analysis further identified that PDW and MLR was independently associated with the risk of early esophageal cancer (both p<0.001). The higher level of NLR (P=0.007) and MLR (P=0.015) were statistically significant with submucosal invasion in EEC patients and the level of MLR were significantly associated with larger tumor size (P=0.030). The results of the validation group were in consistence with the primary group. Conclusions: Hematological parameters of MLR and PDW can be used as an adjuvant tool for the diagnosis of EEC. Moreover, the value of MLR can reflect the invasion depth index.

Differential diagnosis for esophageal protruded lesions using a deep convolution neural network in endoscopic images.

BACKGROUND AND AIMS: Recent advances in deep convolutional neural networks (CNNs) have led to remarkable results in digestive endoscopy. In this study, we aimed to develop CNN-based models for the differential diagnosis of benign esophageal protruded lesions using endoscopic images acquired during real clinical settings. METHODS: We retrospectively reviewed the images from 1217 patients who underwent white-light endoscopy (WLE) and EUS between January 2015 and April 2020. Three deep CNN models were developed to accomplish the following tasks: (1) identification of esophageal benign lesions from healthy controls using WLE images; (2) differentiation of 3 subtypes of esophageal protruded lesions (including esophageal leiomyoma [EL], esophageal cyst (EC], and esophageal papilloma [EP]) using WLE images; and (3) discrimination between EL and EC using EUS images. Six endoscopists blinded to the patients' clinical status were enrolled to interpret all images independently. Their diagnostic performances were evaluated and compared with the CNN models using the area under the receiver operating characteristic curve (AUC). RESULTS: For task 1, the CNN model achieved an AUC of 0.751 (95% confidence interval [CI], 0.652-0.850) in identifying benign esophageal lesions. For task 2, the proposed model using WLE images for differentiation of esophageal protruded lesions achieved an AUC of 0.907 (95% CI, 0.835-0.979), 0.897 (95% CI, 0.841-0.953), and 0.868 (95% CI, 0.769-0.968) for EP, EL, and EC, respectively. The CNN model achieved equivalent or higher identification accuracy for EL and EC compared with skilled endoscopists. In the task of discriminating EL from EC (task 3), the proposed CNN model had AUC values of 0.739 (EL, 95% CI, 0.600-0.878) and 0.724 (EC, 95% CI, 0.567-0.881), which outperformed seniors and novices. Attempts to combine the CNN and endoscopist predictions led to significantly improved diagnostic accuracy compared with endoscopists interpretations alone. CONCLUSIONS: Our team established CNN-based methodologies to recognize benign esophageal protruded lesions using routinely obtained WLE and EUS images. Preliminary results combining the results from the models and the endoscopists underscored the potential of ensemble models for improved differentiation of lesions in real endoscopic settings.

Lesion size and circumferential range identified as independent risk factors for esophageal stricture after endoscopic submucosal dissection.

BACKGROUND AND AIM: Endoscopic submucosal dissection (ESD) is used to treat early esophageal cancer and precancerous lesions. Patients undergoing ESD are prone to esophageal stenosis, which impairs therapeutic efficacy and quality of life. This retrospective study aimed to investigate the potential association between patient demographics and esophageal lesion characteristics with the risk of esophageal stenosis following ESD. METHODS: For this retrospective study 190 consecutive patients who underwent ESD between January 2013 and January 2015 were recruited. Data on patient demographics, esophageal lesion-related factors, operation details, esophageal stenosis occurrence and measures taken to prevent or treat stricture were collected, and the normality of distribution of each indicator was assessed with a Kolmogorov-Smirnov test. Stenosis risk factors were then identified using univariate and multivariate logistic regression. RESULTS: Post-ESD esophageal stenosis occurred in 51 cases. Multivariate logistic regression analysis was performed to identify independent risk factors. A history of EMR/ESD (OR = 4.185, 95% CI: 1.511-11.589), resection circumferential diameter (OR = 1.721, 95% CI: 1.135-2.610), non-en bloc resection (OR = 7.413, 95% CI: 2.398-22.921), submucosal infiltration (OR = 3.449, 95% CI: 1.014-11.734) and circumferential resection range (OR = 57.493, 95% CI: 17.236-191.782) were identified as independent risk factors for post-ESD esophageal stenosis. Spraying porcine fibrin adhesive on the resection bed reduced neither the incidence of postoperative stenosis nor the extent of postoperative dilation. CONCLUSION: Post-ESD esophageal stenosis is significantly related to size and circumferential range of lesion resection. EMR/ESD history, non-en bloc resection and submucosal infiltration may be additional risk factors.

Modified biliary-enteric anastomosis for congenital choledochal cyst: clinical and prognostic analysis of 91 cases.

PURPOSE: To report our experience with a modified biliary-enteric anastomosis procedure for the surgical treatment of congenital choledochal cysts. METHODS: Between January 2009 and December 2013, 91 children (19 boys, 72 girls; ages, 6-145 months) with congenital choledochal cysts were treated with our modified surgical procedure in our hospital. Of these patients, 69 had type I cysts, and 22 had type IV B cysts. The main parameters analyzed mainly included the operative time, duration of bowel recovery, resumption of diet, postoperative hospital stay, liver-function tests, postoperative complications, and prognosis. RESULTS: The average operation duration was 129.34 ± 23.50 min. The time until first flatus and resumption of oral diet were 26.51 ± 4.13 h and 5.47 ± 0.77 day, respectively. The mean postoperative hospital stay was 11.84 ± 2.58 day. Postoperative complications occurred in six patients: intestinal obstruction (1 patient), postoperative bleeding (1 patient), postoperative pancreatitis (1 patient), and bile leakage (3 patients). During a follow-up of 2-7 years, four cases of occasional abdominal pain were found. Contrast agent reflux was detected on upper gastrointestinal imaging in three children. All children had good nutrition. CONCLUSION: The modified biliary-enteric anastomosis is a safe, simple, and reliable technique. However, longer follow-up and a larger sample size are necessary to prove its efficacy in the treatment of congenital choledochal cysts.

A severe course of Cronkhite-Canada syndrome and the review of clinical features and therapy in 49 Chinese patients.

We describe a severe case of Cronkhite-Canada syndrome and review the clinical features and therapy in 49 Chinese patients. A 67-year-old man who underwent severe chronic diarrhea had typical clinical manifestations of hyperpigmentation, hair loss, and dystrophic changes in the fingernails. Although sufficient nutrition support and other therapies reported in the literature were provided, the patient died of systemic failure one year later. Cronkhite-Canada syndrome is characterized by generalized gastrointestinal polyps associated with hyperpigmentation, hair loss, and onycholysis. Anemia, positive stool occult blood, serum electrolyte disturbances, and low serum proteins are the main clinical features of patients with Cronkhite-Canada syndrome. Most patients were diagnosed by esophagogastroduodenoscopy and/or colonoscopy, and polyps were found in the entire alimentary tract, except the notable exception of the esophagus. The polyp-like samples of mucosa, hyperplasia, and adenoma were characterized by acute/chronic inflammation. Four cases were complicated with cancer. The treatment of Cronkhite-Canada syndrome includes symptomatic and support therapy, administration of corticosteroids, antibiotics and acid inhibitors, therapeutic endoscopy, and surgery. While the mortality rate was reported as 47.3% (9/19), some patients may live a long life with controlled symptoms.

NF-kappaB related abnormal hyper-expression of iNOS and NO correlates with the inflammation procedure in biliary atresia livers.

OBJECTIVE: To study the expression of inducible nitric oxide synthase (iNOS) and its related regulators in biliary atresia (BA) livers and exlpore their relationships with the inflammation pathway in BA livers. METHOD: The iNOS expression in livers of 38 cases of BA children, 15 cases of neonatal cholestasis (NC) children, and 18 cases of normal control were, respectively, examined and total nitric oxide (NO) metabolites concentration of all samples were calculated by a colorimetric method based on Griess reaction. The TdT-mediated dupt biotin nick end labeling (TUNEL) method was used to label the apoptotic bile duct epithelial cells and hepatocytes. The western blotting and immunohistochemistry methods to semi-quantitatively analyze the nuclear factor-kappaB (NF-kappaB) expression of each group were also used. RESULTS: The iNOS expression intensity and total NO metabolites concentration of BA group (0.30 +/- 0.08, 90.40 +/- 12.46 micromol/L) were significantly higher than those of NC and normal control groups, P < 0.01. Correlation analysis showed a strong positive correlation between the serum AST level (152.76 +/- 29.59 U/L) and total NO metabolites concentration in BA group. Compared with the NC (32.47 +/- 5.55) and normal control (20.72 +/- 5.63) groups, a significantly higher apoptosis rate of intrahepatic bile duct epithelial cells was found in BA group (54.00 +/- 11.67) that tightly correlated with the iNOS intensity (r = 0.99, P < 0.01). The NF-kappaB intensity of BA group was also significantly higher than that of NC and normal control groups and had a strong positive correlation with the iNOS intensity (r = 0.97, P < 0.01). CONCLUSION: The abnormal hyper-expression of iNOS may play an important role in mediating the inflammation procedure in BA livers. This change perhaps has some relationship with the high expression of NF-kappaB and NO. These regulators can up-regulate the apoptosis of bile duct epithelial cells in BA livers and cause the damage to liver tissues.

Inhibitory effects of somatostatin on cholecystokinin octapeptide induced bile regurgitation under stress: ionic and molecular mechanisms.

OBJECTIVE: To investigate the possible effects and related ionic and molecular mechanisms of changes of plasma cholecystokinin octapeptide and somatostatin on stress-induced bile regurgitation in rats. METHODS: In forty healthy adult rats, changes of plasma cholecystokinin octapeptide, somatostatin and intragastric bile concentration under stressful condition were respectively measured by specific radioimmunoassay methods. Contractile responses of gastric antral smooth strips isolated from healthy adult rats were recorded by polyphysiograph. Immunoprecipitation was used to determine the regulatory effect of protein kinase C on regulating the phosphorylation of type 3 inositol 1,4,5-triphosphate receptor (InsP3R3) in gastric smooth muscle cells. Changes of intracellular calcium fluorescence intensity of smooth muscle cells presented as intracellular calcium ([Ca2+]i) were analyzed under laser scanning confocal microscopy and L-type voltage-dependent calcium currents of smooth muscle cells were recorded by patch-clamp techniques. RESULTS: Compared with the normal control group, plasma cholecystokinin octapeptide and gastric bile concentration of each stress group significantly increased during the stress, while adverse effect was obtained in plasma somatostatin, which decreased from the beginning of the stress and attained the minimum nearly at the same time when the plasma cholecystokinin octapeptide concentration reached the maximum. Respective addition of cholecystokinin octapeptide and somatostatin with increasing concentrations caused rapid, sustained, concentration-dependent increase and decrease in muscle contraction of gastric antral strips, and cholecystokinin octapeptide that increased the contractile response could be blocked by respective administration of nifedipine and somatostatin significantly. Similar results were obtained in the changes of calcium fluorescence intensity and calcium currents of smooth muscle cells. Pretreatment with somatostatin significantly increased cholecystokinin octapeptide-increased phosphorylation of InsP3R3 in smooth muscle cells. CONCLUSIONS: Gastric mucosal damage induced by bile regurgitation is closely connected with gastric antral dysmotility evoked by the changes of cholecystokinin octapeptide and somatostatin under stressful condition. Cholecystokinin octapeptide-intensified contraction depends on the release of intracellular calcium stores and the influx of extracellular calcium via L-type voltage-dependent calcium channels, while this excitatory effect of cholecystokinin octapeptide could be blocked by somatostatin, suggesting that both of the two peptides play important roles in the regulation of gastric motility.

[The role of protein kinase C-mediated phosphorylation of type III inositol 1, 4, 5-triphosphate receptor in cholecystokinin octapeptide induced calcium mobilization in gastric antral smooth muscle cells].

OBJECTIVE: To study the effects of sulfated cholecystokinin octapeptide (sCCK-8S) on intracellular calcium release and extracellular calcium influx in gastric antral smooth muscle cells (SMC) and the mechanism thereof. METHODS: (1) Longitudinal muscle (LM) and circular muscle (CM) strips of gastric antrum and pylorus were isolated from SD rats and suspended in a tissue chamber to record the contractile responses by polyphysiography. (2) Immunoprecipitation, electrophoresis, and immunoblotting were used to detect the phosphorylation of type III inositol 1, 4, 5-triphosphate receptor (InsP(3)R(3)) in the SMCs. (3) The responsiveness of gastric SMC to CCK-8S was examined by using fura-2-loaded microfluorimetric measurement of intracellular calcium concentration ([Ca(2+)]i). (4) The current of L-type calcium channels (ICaL) was recorded by using patch-clamp techniques. RESULTS: (1) Significant changes to CCK-8S were found in the mean contractile amplitude of the CM and frequency of LM of gastric antrum and could be suppressed by CCK-A receptor (CCK-AR) antagonist and ATPase inhibitors. (2) CCK-8S stimulation of SMC resulted in PKC-dependent phosphorylation of the InsP(3)R(3). (3) CCK-8S-evoked significant increase in [Ca(2+)]i [from (69 +/- 7) mol/L to (472 +/- 36) nmol/L, P < 0.01] could be suppressed by CCK-AR antagonist, ATPase inhibitors and protein kinase C (PKC) activator; whereas on condition that extracellular calcium was removed or L-type calcium inhibitor nifedipine was added a small but significant increase of [Ca(2+)]i could be still elicited by CCK-8S. (4) CCK-8S-intensified calcium current [from (-56 +/- 7) pA to (-89 +/- 6) pA, P < 0.01] could be apparently inhibited by respective administration of nifedipine, ATPase inhibitors, and calcium dependent chloride channel (I(Cl-Ca)) blocker (all P < 0.01). CONCLUSION: CCK-8S-evoked [Ca(2+)]i increase in gastric antral SMCs depends on the release of intracellular calcium stores, which is regulated by PKC mediated phosphorylation of InsP(3)R(3). The released intracellular calcium in turn activates the L-type voltage-dependent calcium channels (VDCC) through the activation of calcium dependent chloride channels, and ultimately results in the occurrence of contraction response of smooth muscles.

Reversal effect of thalidomide on established hepatic cirrhosis in rats via inhibition of nuclear factor-kappaB/inhibitor of nuclear factor-kappaB pathway.

BACKGROUND: Suppression of nuclear factor-kappaB (NF-kappaB)/inhibitor of nuclear factor-kappaB (IkappaB) signaling pathway is a potential property of thalidomide. This study was designed to investigate the effects of thalidomide on expressions of NF-kappaB, IkappaB and intercellular adhesion molecule-1 (ICAM-1), and vascular cell adhesion molecule (VCAM-1) in established rat liver cirrhosis. METHODS: Rat liver cirrhosis was achieved by IP injection of carbon tetrachloride (CCl4) three times weekly for 8 weeks. CCl4 was then discontinued and thalidomide (100 mg/kg) or its vehicle was administered daily by gavage for 6 weeks. Hydroxyproline (HYP) content in liver was detected by biochemical assay. NF-kappaBp65, ICAM-1, VCAM-1 and alpha-smooth muscle actin (alpha-SMA) protein in the liver, IkappaBalpha protein in cytoplasm and NF-kappaBp65 protein in nucleus and ICAM-1, VCAM-1 mRNA levels in the liver were studied using immunohistochemistry, Western blot, and reverse transcriptase polymerase chain reaction, respectively. RESULTS: Compared with the spontaneous recovery of cirrhosis, the histopathology of liver of rats given thalidomide was significantly improved. HYP content in liver, the expressions of ICAM-1, VCAM-1 mRNA and protein, NF-kappaBp65 and alpha-SMA protein were decreased significantly and IkappaBalpha protein in liver was elevated significantly in this group. CONCLUSIONS: Thalidomide may exert its effect on downregulation of NF-kappaB-induced adhesion molecules and activation of hepatic stellate cell via inhibition of degradation of IkappaB to reverse established rat hepatic cirrhosis.

Thalidomide prevents rat liver cirrhosis via inhibition of oxidative stress.

This study investigated the effect of thalidomide on oxidative stress in rat liver cirrhosis. The cirrhosis of rat was induced by intraperitoneal injection of carbon tetrachloride thrice weekly; meanwhile, thalidomide (10mg/kg or 100mg/kg) was given daily by intragastric administration for 8 weeks. The content of oxidative stress parameters, including superoxide dismutase, glutathione peroxidase, and malondialdehyde, in the liver was detected by biochemical assay. Immunohistochemistry revealed alpha-smooth muscle actin (alpha-SMA), desmin, and tissue inhibitor of metalloproteinase-1 (TIMP-1) protein in the liver. Nuclear factor kappa B p65 (NF-kappaBp65) protein in nucleus and transforming growth factor beta1 (TGF-beta1) protein in cytoplasm were detected by Western blot. NF-kappaBp65, TGF-beta1, and TIMP-1 mRNA levels in the liver were studied using reverse transcriptase polymerase chain reaction. Liver histopathology was significantly improved in rats given high doses of thalidomide. The content of oxidative stress parameters and the expressions of NF-kappaBp65, TGF-beta1 and TIMP-1 protein, and mRNA were significantly decreased in these animals. The expressions of alpha-SMA and Desmin protein were also significantly decreased in them. Thalidomide might exert an effect on the inhibition of oxidative stress via downregulation of NF-kappaB signaling pathway to prevent the progression of liver cirrhosis.