Objective To investigate the effect of phenylhexyle isothiocyanate (PHI) on demethylation and activation of transcription gene p15 in acute leukemia cell line Molt-4. Methods DNA sequencing and modified methylation specific PCR (MSP) were used to screen p15-M and p15-U mRNA after Moh-4 cells were treated with PHI. P15 mRNA was measured by RT-PCR. Pl5 protein was detected by Western blotting. Results Hypermethylation of gene pl5 was apparently attenuated and activation of transcription p15 gene was de novo after 5 days exposure to PHI. PHI enhanced both the expression of p15 mRNA and p15 protein in a concentration-dependent manner. The ratio of the gray scale of p15 mRNA strap was 0.17±0.12 in control, 0.29±0.14 in PHI 10 μmol/L, 0.55±0.07 in PHI 20 μmol/L, 0.93±0.13 in PHI 40 μmol/L. Conclusion PHI could active demethylation and transcription of gene p15.

Objective To explore the possibility of heterodimerization between orexin type 2α receptor (OX2αR) and orexin type 2β receptor (OX2βR).Methods Using confocal laser scanning microscope,enzyme linked immunosorbent assay (ELISA),fluorescence resonance energy transfer (FRET) and Bioluminescence resonance energy transfer (BRET) to study the interaction between OX2αR and OX2βR.Result Confocal laser scanning microscope and ELISA showed that OX2αR and OX2βR were both expressed in the cytoplasm.The FRET demonstrated that the signal of the experimental group (OX2αR-YFP+ OX2βR-CFP) was significantly stronger than that of control group (YFP+OX2βR-CFP).The BRET value of the experimental group (OX2αR-YFP+OX2βR-Rluc,mBRET ratio was 65± 15) was higher than that of control group (YFP+ OX2βR-Rluc/OX2αR-YFP+Rluc,mBRET ratio was 10±5) (P＜0.05).Conclusion There are heterodimerization between mOX2αR and mOX2βR.

Voltage-gated sodium channels (VGSCs) in primary sensory neurons play a key role in transmitting pain signals to the central nervous system. BmK I, a site-3 sodium channel-specific toxin from scorpion Buthus martensi Karsch, induces pain behaviors in rats. However, the subtypes of VGSCs targeted by BmK I were not entirely clear. We therefore investigated the effects of BmK I on the current amplitude, gating and kinetic properties of Nav1.8, which is associated with neuronal hyperexcitability in DRG neurons. It was found that BmK I dose-dependently increased Nav1.8 current in small-sized (<25 μm) acutely dissociated DRG neurons, which correlated with its inhibition on both fast and slow inactivation. Moreover, voltage-dependent activation and steady-state inactivation curves of Nav1.8 were shifted in a hyperpolarized direction. Thus, BmK I reduced the threshold of neuronal excitability and increased action potential firing in DRG neurons. In conclusion, our data clearly demonstrated that BmK I modulated Nav1.8 remarkably, suggesting BmK I as a valuable probe for studying Nav1.8. And Nav1.8 is an important target related to BmK I-evoked pain.
Aniline Compounds ; pharmacology ; Animals ; Cell Size ; Cells, Cultured ; Electrophysiological Phenomena ; drug effects ; Furans ; pharmacology ; Ganglia, Spinal ; cytology ; Kinetics ; Male ; NAV1.8 Voltage-Gated Sodium Channel ; metabolism ; Rats ; Rats, Sprague-Dawley ; Scorpion Venoms ; antagonists & inhibitors ; pharmacology ; Scorpions ; Sensory Receptor Cells ; drug effects ; metabolism ; physiology ; Sodium Channel Blockers ; pharmacology ; Voltage-Gated Sodium Channel Agonists ; pharmacology

Objective: To study clinical characteristics and treatment after colonscopic perforation, and to determine risk factors for postoperative complications. Methods: Cases diagnosed as colonoscopic perforation within 7 days after colonoscopy in Peking Union Medical College Hospital between January 2010 and January 2017 were reviewed. Data regarding demography (age, sex), clinical information (comorbidities, medication history of glucocorticoid, length of hospital stay), colonoscopy (whether endoscopic therapy or anesthesia was performed, intestinal cleanliness), perforation (region, diagnosing time) and operation (laparotomy or laparoscopic operation, procedure, post-operational complications) were collected. Single factor analysis and Spearman correlation analysis were employed to determine the risk factors of postoperative complications. Results: A total of 14 colonoscopic perforation cases were identified and included in this study, and the overall perforation rate was 0.03%. Most perforations occurred in rectum (2 cases) and sigmoid colon (8 cases). Twelve perforation patients received operational treatment, of who 6 developed postoperative complications, including 3 cases of incision infection, 2 cases of peritoneal infection, 1 case of catheter-related infection and 1 case of pulmonary embolism. Spearman correlation analysis showed that preoperative medication of glucocorticoid and non-rectosigmoid perforation were positively related to postoperative complications (both correlation coefficients were 0.707, P=0.01), while perforation diagnosed immediately and satisfying intestinal cleanliness were negatively related to it (both correlation coefficients were -0.667, P<0.05). Conclusion Perforations are rare but severe complications of colonoscopy, and surgical interventions are necessary in most cases. Postoperative complications were significantly related to perforation sites, preoperative medication of glucocorticoid, perforation diagnosis time and intestinal cleanliness.

Objective:To explore the endoscopic features of early gastric cancer (EGC) related to non-curative endoscopic resection, and to construct an assessment model to quantify the risk of non-curative resection.Methods:From August 2006 to October 2019, 378 lesions that underwent endoscopic resection and were diagnosed pathological as EGC in the Department of Gastroenterology, Peking Union Medical College Hospital were included in this case-control study.Seventy-eight (20.6%) non-curative resection lesions were included in the observation group, and 234 lesions which selected from 300 lesions of curative resection were included in the control group according to the difference of operation year ±1 with the observation group, and the ratio of 1∶3 of the observation group to the control group. Univariate and multivariate logistic regression analysis were performed to explore the risk factors for non-curative resection. The independent risk factor with the minimum β coefficient was assigned 1 point, and the remaining factors were scored according to the ratio of their β coefficient to the minimum. A predictive model was established to analyze the 378 lesions.The non-curative resection rates of lesions of different scores were calculated. Results:Univariate analysis showed that the lesion diameter, the location, redness, ulcer or ulcer scar, fold interruption, fold entanglement, and invasion depth observed with endoscopic ultrasonography (EUS) were associated with non-curative resection of EGC lesions ( P<0.05), and contact or spontaneous bleeding may be associated with non-curative resection ( P=0.068). Multivariate logistic regression analysis showed that submucosal involvement (VS confined to the mucosa: β=0.901, P=0.011, OR=2.46, 95% CI: 1.23-4.92), lesion diameter of 3-<5 cm (VS <3 cm: β=0.723, P=0.038, OR=2.06, 95% CI: 1.04-4.09), lesion diameter of ≥5 cm (VS <3 cm: β=2.078, P=0.003, OR=7.99, 95% CI: 2.02-31.66), location in the upper 1/3 of the stomach (VS lower 1/3: β=1.540, P<0.001, OR=4.66, 95% CI: 2.30-9.45), and fold interruption ( β=2.287, P=0.008, OR=1.93, 95% CI: 0.95-3.93) were independent risk factors for non-curative resection of EGC lesions. The factor of lesion diameter of 3-<5 cm and submucosal involvement were assigned 1 point respectively, location in the upper 1/3 of the stomach was assigned 2 points, diameter of ≥5 cm and fold interruption were assigned 3 points respectively, and other factors were assigned 0 point. Then the analysis of 378 lesions showed that the probability of non-curative resection at ≥2 points was 41.9% (37/93), 4 times as much as that at 0 [11.5% (25/217)]. Conclusion:EGC lesions with diameter ≥3 cm, located in the upper 1/3 of the stomach, interrupted folds or submucosal involvement are highly related to non-curative resection. The predictive model based on these factors achieves satisfactory efficacy, but it still needs further validation in larger cohorts.

Brain renin-angiotensin system (RAS) is closely associated with many pathophysiological processes of cardiocerebrovascular diseases,including stroke.The activation of the different components in RAS will produce specific biological effects.This article reviews the roles of brain RAS in the pathophysiological processes of ischemic stroke,especially the neuroprotective effect of ACE2/Ang-(1-7)/Mas axis.

Objective To reinforce the awareness of colorectal endometriosis (EM) in colonoscopy examination.Methods Patients diagnosed as colorectal EM at Peking Union Medical College Hospital between February 2002 and February 2017 were enrolled in this study.The clinical characteristics and endoscopic features of EM lesions were summarized and compared between pathologically positive group and negative group.Results A total of 34 cases were included with average age of (38.3± 8.9) years old.All EM lesions located within rectum and sigmoid colon.The endoscopic lesions manifested as protrusion in 21 cases (61.8％) and protrusion-depression in 13 cases (38.2％),local stenosis in 8 cases (23.5％);erosive surface in 33 cases (97.1％) with local spontaneous hemorrhage in 4 cases (11.8％);nodal surface in 23 cases (67.6％),and lymphangiectasis base in 9 cases (26.4％).Endoscopic biopsy specimens were obtained in all cases with average 3 (2,4) pieces.Positive results were found only in 4 patients (11.8％) with 3 endometriosis and one (endometrial) adenosarcoma.Compared with negative group,spontaneous hemorrhage was more frequent in positive group (2/4 vs.2/30,P=0.013).Mean biopsy sample number was significantly larger in positive group (5 vs.3,P=0.004).Conclusions Colorectal endometriosis is mostly located within rectosigmoid region.Endoscopic features mainly include protrusion or protrusion-depression lesions with erosive and nodular surface,or local stenosis.Spontaneous hemorrhage under colonoscopy yields higher positive rate for biopsy,thus increasing biopsy sample numbers may improve pathology results.

Objective:To summarize the clinical characteristics of bleeding during endoscopic submucosal dissection (ESD) and to analyze the risk factors for bleeding.Methods:Data of patients who received gastric ESD in endoscopy center of Peking Union Medical College Hospital from January 2015 to December 2019 were reviewed. The medical history, characteristics of gastric lesions, operation process and prognosis of the patients were analyzed.Results:A total of 437 gastric lesions of 422 patients were included in this study, and 406 lesions were gastric epithelial tumors. The bleeding rate during ESD was 32.3% (141/437), including 2 cases of acute massive hemorrhage. Intraoperative hemorrhage during ESD increased the incidence of myometrial injury and intraoperative perforation, and prolonged the operation time. Multivariate regression analysis showed that risk factors for bleeding during ESD were anatomical adhesion, the lesion location in the upper and middle 2/3 of the stomach, the lesion area ≥ 15 cm 2, male, and non-ESD absolute indications. Conclusion:Bleeding is the speed limiting factor for gastric ESD. For male patients, when the lesion is located in the upper and middle 2/3 of the stomach, large with anatomical adhesion during operation, precaution should be taken for intraoperative hemorrhage.

Objective:To investigate the expression at protein level and diagnostic value of histone acetyltransferase MYST2 in pancreatic cancer.Methods:From December 1st, 2017 to June 30th, 2020, at Peking Union Medical College Hospital, a total of 54 cases of pancreatic cancer tissues and corresponding paracancerous pancreatic tissues (>5 cm from the surgical margin) resected and confirmed by pathology were collected. ASPC1 and BXPC3 pancreatic cancer cell lines were knocked down (ASPC1 and BXPC3 knockdown group), CFPAC1 and SW1990 pancreatic cancer cell lines were overexpressed (CFPAC1 and SW1990 overexpression group), the untreated ASPC1, BXPC3, CFPAC1 and SW1990 were set as blank vector control group. The expression at protein level of MYST2 was detected by Western blotting in patients with different degrees of pathological differentiation, human normal pancreatic duct epithelial cell line HPDE, human pancreatic cancer cell lines ASPC1, BXPC3, CFPAC1 and SW1990, knockdown group, overexpression group and blank vector control group. The cell proliferation, migration, invasion and colony formation ability of the knockdown group, overexpression group and blank vector control group were determined by real-time cellular analysis, Transwell migration and invasion test, and plate colony formation assay. MYST2 immunohistochemical scoring was performed on pancreatic cancer tissues and para cancer tissues. Receiver operating characteristic curve was drawn to analyze the value of different MYST2 protein expression levels in the diagnosis of pancreatic cancer. Independent sample t test and variance analysis were used for statistical analysis. Results:Among the pathological slides of 54 cases of pancreatic cancer, 13 cases were highly differentiated, 24 cases were moderately differentiated, 15 cases were poorly differentiated and 2 cases were undifferentiated, the MYST2 expression at protein level in pancreatic cancer cells was 3.12±1.67, 2.87±1.59, 2.12±1.03 and 1.08±0.34, respectively, and the difference was statistically significant ( F=1.241, P<0.05). The MYST2 expression levels of ASPC1, BXPC3, CFPAC1 and SW1990 were all higher than that of normal pancreatic ductal epithelial cell lines HPDE (1.41±0.47, 1.40±0.93, 1.13±0.62 and 1.71±0.46 vs. 0.82±0.25), and the differences were statistically significant( t=1.625, 1.577, 1.319 and 1.832, all P<0.05). The MYST2 expression level of BXPC3 knockdown group was lower than that of BXPC3 blank vector control group (0.39±0.12 vs. 0.75±0.34); that of ASPC1 knockdown group was lower than that of ASPC1 blank vector control group (0.43±0.22 vs. 0.82±0.48); that of CFPAC1 overexpression group was higher than that of CFPAC1 blank vector control group (1.38±0.45 vs. 0.82±0.37); that of SW1990 overexpression group was higher than that of SW1990 blank vector control group (1.34±0.65 vs. 0.51±0.22), and the differences were statistically significant ( t=1.414, 1.378, 1.319 and 1.934, all P<0.05). The cell proliferation of ASPC1 knockdown group was slower than that of ASPC1 blank vector control group, and the proliferation peak at 80 h was lower than that of blank vector control group (1.02±0.77 vs. 4.31±2.45); the cell proliferation of BXPC3 knockdown group was slower than that of BXPC3 blank vector control group, and the proliferation peak at 80 h was lower than that of blank vector control group (0.91±0.24 vs. 2.84±0.53); the proliferation of pancreatic cancer cells in SW1990 overexpression group was faster than that of SW1990 blank vector control group, and the proliferation peak at 80 h was higher than that of blank vector control group (3.10±0.67 vs. 1.04±0.17); the proliferation of pancreatic cancer cells in CFPAC1 overexpression group was faster than that that of CFPAC1 blank vector control group, and the proliferation peak at 80 h was higher than that of blank vector control group (5.45±1.13 vs. 1.01±0.29), and the differences were statistically significant ( t=1.427, 1.316, 1.292 and 1.501, all P<0.05). In the test of migration ability, the number of cells passed through the Transwell chamber of ASPC1 knockdown group was less than that of ASPC1 blank vector control group (34.08±17.62 vs. 118.76±5.31); that of BXPC3 knockdown group was less than that of BXPC3 blank vector control group (18.62±9.64 vs. 57.90±12.67); that of SW1990 overexpression group was more than that of SW1990 blank vector control group (134.84±24.65 vs. 37.82±6.73); that of CFPAC1 overexpression group was more than that of CFPAC1 blank vector control group (65.79±27.46 vs. 11.68±5.13), and the differences were statistically significant ( t=1.475, 1.322, 1.437 and 1.219, all P<0.05). In the test of invasion ability, the number of cells passed through the Transwell chamber of ASPC1 knockdown group was less than that of ASPC1 blank vector control group (9.79±5.75 vs. 45.76±12.71); that of BXPC3 knockdown group was less than that of BXPC3 blank vector control group (23.46±11.13 vs. 84.92±17.65); that of SW1990 overexpression group was more than that of SW1990 blank vector control group (156.42±34.50 vs. 42.13±22.17); that of CFPAC1 overexpression group was more than that of CFPAC1 blank vector control group (112.64±47.82 vs. 39.09±17.23), and the differences were statistically significant ( t=1.324, 1.635, 1.423 and 1.119, all P<0.05). The number of colony formation of the ASPC1 knockdown group was less than that of ASPC1 blank vector control group (13.15±6.42 vs. 86.79±35.17); that of BXPC3 knockdown group was less than that of BXPC3 blank vector control group (14.93±9.30 vs. 52.93±15.76); that of SW1990 overexpression group was more than that of SW1990 blank vector control group (129.10±57.31 vs. 62.42±37.43); that of CFPAC1 overexpression group was more than that of CFPAC1 blank vector control group (157.98±66.45 vs. 74.35±34.69), and the differences were statistically significant ( t=1.148, 1.290, 1.274 and 1.462, all P<0.05). The MYST2 score of pancreatic cancer tissues was higher than that of adjacent paracancerous pancreatic tissues (3.04±2.23 vs. 1.32 ± 0.70), and the difference was statistically significant ( t=3.479, P<0.05). When the total immunohistochemistry score of MYST2 was 3 point, the area under the curve was the largest (0.888, 95% confidence interval 0.827 to 0.948), and the Youden index was 0.56. Conclusion:MYST2 is associated with the proliferation, invasion and migration of pancreatic cancer cells, and promotes the development of pancreatic cancer.

Objective:To investigate the role of endoscopic ultrasonography (EUS) in differentiating between autoimmune pancreatitis (AIP) and pancreatic cancer (PC).Methods:Data of 133 patients with AIP and 113 patients with PC who underwent EUS because of obstructive jaundice at Peking Union Medical College Hospital from January 2013 to December 2018 were retrospectively analyzed in the study, and were randomly divided into either a derivation sample or a validation sample using 1∶1 allocation according to the random number. In the derivation sample, 10 EUS characteristics were used to construct a prediction model to distinguish between AIP and PC, in which predictors were identified by multivariate stepwise logistic regression analysis and predictive efficacy was evaluated by receiver operating characteristics (ROC) curve analysis. The predictive efficacy was assessed in the validation sample. In view of the subjectivity in the judgment of diffuse/focal hypoechogenicity, 2 prediction models were designed in order to avoid bias.Results:By multivariate stepwise logistic regression analysis, diffuse hypoechogenicity ( OR=591.0, 95% CI: 98.8->999.9, P<0.001) and vessel involvement ( OR=11.9, 95% CI: 1.4-260.2, P=0.023) were identified as statistically significant predictors for distinguishing AIP from PC. EUS characteristics excluding diffuse/focal hypoechogenicity were stepped by logistic regression, which showed that hyperechoic foci/strands ( OR=177.3, 95% CI: 18.7->999.9, P<0.001), pancreatic duct dilation ( OR=60.5, 95% CI: 6.2->999.9, P=0.004), bile duct wall thickening ( OR=35.4, 95% CI: 3.7->999.9, P=0.009), lymphadenopathy ( OR=16.8, 95% CI: 1.7-475.2, P=0.038) and vessel involvement ( OR=22.7, 95% CI: 2.0-725.7, P=0.028) were statistically significant predictors to distinguish the two diseases. Both prediction models were built in the derivation sample, with area under the ROC curve of 0.995 and 0.979 respectively. In the validation sample, sensitivity, specificity, accuracy, positive predictive value and negative predictive value of both prediction models were all >90% by using the optimal cutoff value. Even for discrimination between focal AIP and PC, sensitivity and accuracy of both models were >90%, and specificity, positive predictive value and negative predictive value were all >85%. Conclusion:The 2 prediction models have good differential predictive value, and EUS is a useful tool to differentiate between AIP and PC.