AIM:To explore the effect and mecha-nism of ethanol extract of Angelica sinensis(Oliv.)Diels(EEA)on cell proliferation and apoptosis in B16-F10 melanoma cells.METHODS:Cell viability was analyzed by MTT method.Cell proliferation was detected by colony formation assay.The invert-ed microscope was used to observe the changes of cell growth confluence and morphology.Hoechst 33342 staining was used to detect cell apoptosis.Flow cytometry(FCM)was used to detect cell cycle and apoptosis.Transmission electron microscopy(TEM)was used to observe the changes of cell mi-tochondrial structure.Western blot was used to de-tect the expression levels of cell cycle,apoptosis,mitochondrial biogenesis,and mitochondrial dy-namics-related proteins.RESULTS:Compared with the blank control group,the cell viability of B16-F10 melanoma cells was reduced after EEA(10-400μg/mL)treatment for 24 h and 48 h,respectively(P＜0.05,P＜0.01).The decreased cell growth conflu-ence,morphological changes such as shrinkage,rounding,and reduction in the volume,and apop-totic morphologic changes such as chromatin con-densation were observed after EEA(100 μg/mL and 200 μg/mL)treatment for 24 h.The number of cell clones was decreased after EEA(10-200 μg/mL)treatment for 14 d(P＜0.01).The morphology of mitochondria became more round and shorter,and the inner mitochondrial matrices were either damaged or absent after 200 μg/mL EEA treatment for 24 h.The ratio of cells in G0/G1 phase and the early apoptosis rate of cells were higher than those of the blank control group(P＜0.01)after EEA(20-200 μg/mL)treatment for 24 h.Western blot re-sults showed that compared with the blank control group,the protein expression levels of cleaved cas-pase-9,Bax,DRP1,and FIS1 were up-regulated(P＜0.05,P＜0.01),and the protein expression levels of cyclin D1,cyclin E,CDK2,CDK4,Bcl-2,Bad,Bcl-XL,SIRT1,PGC-1α,NRF1,TFAM,MFN2,and OPA1 were down-regulated(P＜0.05,P＜0.01).CONCLUSION:EEA has an inhibitory effect on the proliferation of B16-F10 melanoma cells,which may be related to the induction of G1/S cell cycle arrest and mito-chondrial apoptotic pathway,and the disruption of mitochondrial biogenesis and mitochondrial dy-namics.

AIM:To explore the effect and mecha-nism of ethanol extract of Angelica sinensis(Oliv.)Diels(EEA)on cell proliferation and apoptosis in B16-F10 melanoma cells.METHODS:Cell viability was analyzed by MTT method.Cell proliferation was detected by colony formation assay.The invert-ed microscope was used to observe the changes of cell growth confluence and morphology.Hoechst 33342 staining was used to detect cell apoptosis.Flow cytometry(FCM)was used to detect cell cycle and apoptosis.Transmission electron microscopy(TEM)was used to observe the changes of cell mi-tochondrial structure.Western blot was used to de-tect the expression levels of cell cycle,apoptosis,mitochondrial biogenesis,and mitochondrial dy-namics-related proteins.RESULTS:Compared with the blank control group,the cell viability of B16-F10 melanoma cells was reduced after EEA(10-400μg/mL)treatment for 24 h and 48 h,respectively(P＜0.05,P＜0.01).The decreased cell growth conflu-ence,morphological changes such as shrinkage,rounding,and reduction in the volume,and apop-totic morphologic changes such as chromatin con-densation were observed after EEA(100 μg/mL and 200 μg/mL)treatment for 24 h.The number of cell clones was decreased after EEA(10-200 μg/mL)treatment for 14 d(P＜0.01).The morphology of mitochondria became more round and shorter,and the inner mitochondrial matrices were either damaged or absent after 200 μg/mL EEA treatment for 24 h.The ratio of cells in G0/G1 phase and the early apoptosis rate of cells were higher than those of the blank control group(P＜0.01)after EEA(20-200 μg/mL)treatment for 24 h.Western blot re-sults showed that compared with the blank control group,the protein expression levels of cleaved cas-pase-9,Bax,DRP1,and FIS1 were up-regulated(P＜0.05,P＜0.01),and the protein expression levels of cyclin D1,cyclin E,CDK2,CDK4,Bcl-2,Bad,Bcl-XL,SIRT1,PGC-1α,NRF1,TFAM,MFN2,and OPA1 were down-regulated(P＜0.05,P＜0.01).CONCLUSION:EEA has an inhibitory effect on the proliferation of B16-F10 melanoma cells,which may be related to the induction of G1/S cell cycle arrest and mito-chondrial apoptotic pathway,and the disruption of mitochondrial biogenesis and mitochondrial dy-namics.

The ageing population has made intestinal ischemia more common in clinical practice.The clinical manifestations of intestinal ischemia caused by various pathological conditions range from mild to non-specific symptoms to life-threatening.Accurate judgement in the early stages of the disease can be life-saving.This article provides an overview of the anatomical and pathophysiological basis for the occurrence of intestinal ischemia, as well as the latest evidence-based medical evidence for the diagnosis and treatment of this condition.The aim is to raise clinicians' awareness of intestinal ischemic injury in the elderly, promote standardized diagnosis and management, and improve clinical outcomes.

The ageing population has made intestinal ischemia more common in clinical practice.The clinical manifestations of intestinal ischemia caused by various pathological conditions range from mild to non-specific symptoms to life-threatening.Accurate judgement in the early stages of the disease can be life-saving.This article provides an overview of the anatomical and pathophysiological basis for the occurrence of intestinal ischemia, as well as the latest evidence-based medical evidence for the diagnosis and treatment of this condition.The aim is to raise clinicians' awareness of intestinal ischemic injury in the elderly, promote standardized diagnosis and management, and improve clinical outcomes.

Objective:To evaluate the clinical efficacy and safety of endoscopic ultrasound (EUS)-guided coil implantation combined with glue injection for preventing rebleeding of tumorous gastric fundal varices.Methods:A retrospective analysis was performed on the clinical data of patients who underwent EUS-guided coil implantation combined with glue injection due to tumorous gastric fundal variceal bleeding from September 2017 to December 2020 in the First Affiliated Hospital, Zhejiang University School of Medicine. The basic characteristics and endoscopic manifestations of patients treated with this technique were analyzed, and the success rate of this technique, the usage of tissue glue and coil, the variceal obliteration rate and the incidence of complications such as pulmonary embolism were analyzed. Patients were followed up over a long period of time to obtain data on rebleeding. At the same time, patients were divided into complete obliteration group and incomplete obliteration group according to the endoscopic performance after the treatment, and the rebleeding rate in the two groups was compared.Results:Among the 41 patients included in this study, 18 cases (43.90%) had single isolated gastric varices type 1, and 23 cases (56.10%) had gastroesophageal varices combined. Fourteen cases had signs of recent bleeding such as thrombi and ulcers under endoscopy, and 13 cases had combined portal hypertensive gastropathy. All patients successfully completed the operation of EUS-guided coil implantation combined with glue injection. The technical success rate for the first attempt was 92.68% (38/41). The one-time pre-installation success rate of coils was 95.12% (39/41). The success rate of target vessel aspiration under EUS guidance was 100.00% (41/41). The release success rate of coil was 97.56% (40/41). The number of coils used was 1.12±0.33, and the amount of tissue glue used after inserting the coils was 1.54±0.46 mL. The target vessel obliteration rate after operation was 97.56% (40/41), and the complete gastric fundal variceal obliteration rate was 51.22% (21/41). There were 2 cases (4.88%) of intraoperative bleeding, 1 case stopped spontaneously, and 1 case successfully stopped bleeding after additional injection of tissue glue under direct endoscopic vision. No patient had symptoms of pulmonary embolism such as decreased oxygen saturation during the operation. Postoperative computed tomography angiography of pulmonary artery or lung computed tomography was performed in 24 cases, of which 1 case (4.17%) showed distal pulmonary artery embolism, but there were no relevant clinical manifestations. The patients were followed up for 21.64±16.86 months (1-50 months) after the surgery. There were 13 cases (31.71%) of recurrent gastrointestinal bleeding after the surgery, no rebleeding within 5 days, 2 cases of rebleeding within 3 months, 5 cases from 3 to 12 months, and 6 cases more than 1 year. Among the 13 cases of recurrent bleeding after the surgery, the recurrence rate of bleeding after operation in the complete obliteration group was 23.81% (5/21), which was lower than that in the incomplete obliteration group with 40.00% (8/20), but there was no statistically significant difference ( χ2=1.240, P=0.265). Conclusion:EUS-guided coil implantation combined with glue injection has good secondary prevention value and safety for tumorous gastric fundal variceal bleeding, and is worthy of clinical promotion and application.

Objective:To investigate the clinical characteristics of adult eosinophilic esophagitis (EoE) and the diagnostic value of endoscopic ultrasonography (EUS).Methods:Data from 12 adult patients with EoE diagnosed at the First Affiliated Hospital of Zhejiang University School of Medicine from 2011 to 2022 were retrospectively analyzed for the clinical manifestations, imaging and endoscopic findings, EUS characteristics, treatment and prognosis.Results:Among the 12 EoE patients, 9 were male and 3 were female, with a mean age at diagnosis of 48.5 years (17-73 years). Two patients had a history of asthma, 1 had urticaria, and 5 had a history of food and/or drug allergy. The time from symptom onset to diagnosis ranged from 2 weeks to 30 years. The most common clinical symptoms were acid regurgitation (8 cases), abdominal pain (8 cases), and obstruction dysphagia (5 cases). Four (4/11) patients had increased peripheral blood eosinophils and 4 (4/10) patients had increased peripheral blood immunoglobulin E. Contrast-enhanced CT of the esophagus showed esophageal wall thickening and enhancement in 7 patients (7/8). One (1/8) patient showed multiple mediastinal lymph nodes enlargement. Under endoscopy, edema (12 cases), linear furrows (5 cases), rings (4 cases), exudates (1 case), and luminal narrowing (1 case) were observed in the mucosa. Eight patients underwent EUS, and all of them showed esophageal wall thickening, and the muscularis propria thickening being the most common (5.0-12.5 mm), with various hierarchical structures and unclear boundaries. Topical corticosteroid therapy led to significant symptom improvement and mucosal healing in 6 patients. Biopsy showed that the infiltration of eosinophils in the esophageal mucosa was significantly reduced or completely relieved. The clinical symptoms recurred in 5 patients after receiving acid suppression therapy, and 2 of them still had eosinophil infiltration in esophageal mucosal.Conclusion:The clinical presentation of EoE in adults can mimic gastroesophageal reflux disease, highlighting the importance of accurate diagnosis. The typical endoscopic findings of EoE are edema, linear furrows, rings, exudates and luminal narrowing. EUS can be used for the diagnosis, differential diagnosis, and objective evaluation of esophageal wall remodeling in patients with EoE. Adult patients with EoE have poor response to antacid therapy alone, while topical corticosteroid therapy can achieve good clinical efficacy and pathological remission.

Objective: To evaluate the efficacy and safety of Oryz-Aspergillus enzyme and pancreatin tablets (Combizym^®) in the treatment of postprandial distress syndrome (PDS) in the elderly, compared with gastrointestinal motility drugs. Methods: A prospective randomized controlled trial was designed and registered in the China Clinical Trials Registry (ChiCTR-IPR-16008185). The elderly patients with PDS were randomly divided into three groups, including Mosapride group with Mosapride citrate tablets 5 mg 3 times per day for 2 weeks; Combizym^® group with Combizym tablets 244 mg 3 times per day for 2 weeks; combined treatment group with both drugs and same doses for 2 weeks. The modified Nepean dyspepsia index (NDSI) score, discomfort intensity score and PDS score were calculated on patients before treatment, at the end of first and second week of treatment, as well as 4 weeks after treatment finished, respectively. Adverse effects were evaluated. Results: A total of 323 patients from 16 tertiary hospitals in China were enrolled in this study. Among them, 105 patients were in Mosapride group, 109 in Combizym^® group and 109 in combined treatment group. There were 148 males (45.8%) and 175 females (54.2%) with median age 71.4±9.0 years (60-100 years). Baseline characteristics of three groups were comparable. After treatment, the NDSI scores in three groups all decreased significantly (P<0.001), while they were similar between groups (P>0.05). The discomfort intensity score and PDS score in three groups showed a significant reduction after treatment (P<0.001), especially in the combined treatment group. Compared with Mosapride group, the scores in Combizym^® group decreased significantly after one or two weeks [discomfort intensity score: after one week, 4.0(2.5, 8.0) vs. 6.0(3.0, 10.0); after two weeks, 3.0(0.0, 5.0) vs. 4.0(2.0, 6.0); all P<0.05. PDS score: after one week, 6.0(3.0, 9.0) vs. 7.0(3.5, 10.5); after two weeks, 3.0(0.0, 5.0) vs. 4.0(2.0, 7.0); all P<0.05]. The efficacy rate in all patients after first week of treatment was over 15.0%. The efficacy rates after two weeks were 55.2%, 68.8% and 73.4% in Mosapride group, Combizym^® group and combined treatment group, respectively. After two week treatment, the efficacy rates in Combizym^® group (P=0.041) and combined group (P=0.006) were higher than that of Mosapride group. The recurrence rate of Mosapride group was 9.5%, which was significantly higher than that of Combizym^® group (1.8%, P<0.05) and combined treatment group (1.8%, P<0.05). There were no serious adverse effects in the three groups. Conclusions The efficacy of Oryz-Aspergillus enzyme and pancreatin tablets is comparable with that of Mosapride in elderly PDS patients, with fewer adverse effects and low recurrence rate. Combination regimen indicates better efficacy than that of Oryz-Aspergillus enzyme and pancreatin tablets or Mosapride alone.

Objective:To explore the diagnostic value of endoscopic ultrasonography (EUS) for duodenal accessory papilla.Methods:Data of 122 cases of duodenal accessory papilla diagnosed by EUS at the endoscopy center of the First Affiliated Hospital of Zhejiang University School of Medicine from February 28, 2006 to February 28, 2018 were analyzed and summarized.Results:Of the 122 duodenal accessory papilla cases, the age was 52.1±12.9, with more males than females. The most common site of duodenal accessory papillae was the descending part above the papilla (88/122, 72.13%), followed by the junction of duodenal bulb and descending part (29/122, 23.77%), and a small proportion of lesions located in the duodenal bulb (5/122, 4.10%). Duodenal accessory papillae were all solitary, whose diameter mostly ranged 0.5-1.0 cm (88/122, 72.13%), a smaller proportion of diameter larger than 1.0 cm (23/122, 18.85%), and only a few with diameter less than 0.5 cm (11/122, 9.02%). Most duodenal accessory papillae were hypoechoic (71/122, 58.20%) or moderate to low echogenic (35/122, 28.68%), and the echoes were mostly homogeneous. The mucosa layer was smooth, with a sphincteroid structure in the submucosa and below. The boundary of the duodenal accessory papillae was mostly clear (121/122, 99.18%) and characteristic lacunar cavity structures were often seen in the center (83/122, 68.03%). The surrounding intestinal wall was normal and no associated enlarged lymph nodes were found around the intestine.Conclusion:EUS can clearly show the structure of duodenal accessory papilla and adjacent organs, and is of high value for the diagnosis of duodenal accessory papilla.

Objective: To screen and identify serum protein biomarkers for the differential diagnosis between ischemic colitis (IC) and ulcerative colitis (UC) by tandem mass tag (TMT) combined with liquid chromatography/tandem mass spectrometry (LC-MS/MS). Methods: From January 2018 to January 2019, at the First Affiliated Hospital of School of Medicine of Zhejiang University, patients with UC or IC, and health controls, each 10 cases, were enrolled into UC group, IC group and normal control (NC) group, respectively. Fasting serum samples of all the subjects were collected. After removal of high-abundance protein, followed by proteolysis, peptide labeling and fractionating, the samples were then processed by mass spectrometry. The protein with TMT data of three groups was obtained and protein with TMT value 0 were removed. Heat map of protein was constructed. The differential protein was defined as the protein fold change over 1.5 or less than 0.67. The Reactome database was used to cluster the pathways of differential proteins among groups. Statistical methods included t test, hypergeometry test and corrected by BH multiple test. Results: A total of 357 serum proteins were identified by proteomic profiling. There were 27 differential proteins between the IC group and the NC group, including six up-regulated proteins and 21 down-regulated proteins. There were 228 differential proteins between the UC group and the NC group, including 75 up-regulated proteins and 153 down-regulated proteins. There were 49 differential proteins between UC group and IC group, including 22 up-regulated proteins and 27 down-regulated proteins. In the comparison of differential proteins between the NC group, IC group and UC group, only the expression of fibrin 3 was statistically significant (the fold change between UC and NC, between UC and IC, between IC and NC were 0.24, 0.46 and 0.53, respectively; t=-5.089, -7.298 and -3.919, all P<0.01). The results of pathway cluster analysis showed that in the comparison of differential proteins between IC group and NC group, only the platelet degranulation pathway was enriched, and 10 proteins were involved in this pathway (P<0.01). In the comparison of differential proteins between UC group and NC group, there were 58 pathways enriched, of which 38 proteins were involved in the platelet degranulation pathway, 16 proteins were involved in the initial complement trigger pathway, 13 proteins were involved in the complement cascade pathway, and 11 proteins were involved in antibody-mediated complement activation pathway (all P<0.01). In the comparison of differential proteins between UC group and IC group, three different pathways were obtained. Among them, nine proteins were involved in the platelet degranulation pathway, seven proteins were involved in the initial complement trigger pathway, and five proteins were involved in the complement cascade pathway (all P<0.01). Conclusions The difference in serum proteome between IC patients and UC patients was significant, and the differential proteins are mainly involved in platelet activation and complement activation. The candidate proteins identified in this study may be used as biomarkers for the differential diagnosis of UC and IC in the future.

Objective To set up a computer-assisted polyp detection system under colonoscopy,and to preliminarily verify its effectiveness.Methods Based on Faster R-CNN algorithm and the open source implementation of the open source framework tensorflow and Faster R-CNN,a computer-assisted polyp detection system under colonoscopy was constructed.According to the size and difficulty of the training set,five test groups were set up:test group one,two,three and four contained 1 000,2 000,4 000 and 6 000 training samples,respectively.Test group five increased the probability of selecting the difficult samples based on 6 000 training samples.In different training sets,the sensitivity,specificity,other classification evaluation parameters,and the evaluation parameters of target detection such as recall and precision of this polyps detection system were calculated.Results Classification evaluation parameters showed that the sensitivities of test group one,two,three,four and five were 90.1％,93.3％,93.3％,93.3 ％ and 93.5 ％,respectively,and the difference was statistically significant (x2 =25.324,P＜0.01).The sensitivities of test group two,three,four and five were all higher than that of test group one,and the differences were statistically significant (x2 =13.964,13.508,13.508 and 13.386,all P＜ 0.006 25).There were no significant differences in specificity and positive predictive value among test groups (both P＞0.05).The negative predictive values of test group one,two,three,four and five were 90.4％,93.3％,93.3％,93.3％ and 93.5％,respectively,and the differences were statistically significant (x2 =21.862,P＜0.01).The negative predictive values of test group two,three,four and five were higher than that of test group one,and the differences were statistically significant (x2=11.447,11.564,11.755,13.760;all P＜0.006 25).As the training sample size increased from 1 000 to 2 000,the area under curve (AUC) increased by 2％,and further increased the sample size to 6 000,AUC increased by less than 1 ％.At this point maintaining the same sample size while increasing the proportion of difficult samples,AUC increased by 0.4％.The results of evaluation parameters of target detection showed that the recall rate of each test group was 73.6％,79.8％,79.5％,79.8％ and 83.3％,respectively,and the differences were statistically significant (x2 =71.936,P＜0.01).Among them,the recall rates of test group two,three and four were higher than that of test group one,and the differences were statistically significant (x2 =25.960,23.492 and 25.960,all P＜0.006 25),and the recall rate of test group five was higher than those of test group one,two,three and four,and the differences were statistically significant (x2=67.361,9.899,11.527 and 9.899;all P＜0.006 25).In addition,the precision rates of test group one,two,three,four and five were 87.9％,85.3％,90.2％,91.4％ and 89.2％,respectively,and the difference was statistically significant (x2=48.194,P＜0.01).The precision rates of test group three and five were higher than that of test group two,and the differences were statistically significant (x2 =24.508 and 15.223,both P＜0.006 25),and the precision rate of test group four was higher than those of test group one and two,and the differences were statistically significant (x2=13.524 and 39.120,both P＜0.006 25).As samples size and training difficulty increased,the values of F1-score and mean average precision increased steadily.Conclusions This study initially constructed a computer-assisted polyp detection system under colonoscopy.Currently the maximum sensitivity reached 93.5％,and the maximum recall rate reached 83.3％.Increasing the training set size may improve the polyp detection result to a certain degree,however it will reach a bottleneck.At this time,increasing the training difficulty can further improve the detection scores,especially the recall rate.