1.Establishment of an artificial intelligence assisted diagnosis model based on deep learning for recognizing gastric lesions and their locations under gastroscopy in real time
Xian GUO ; Ying-Yang WU ; Ai-Rui JIANG ; Chao-Qiang FAN ; Xue PENG ; Xu-Biao NIE ; Hui LIN ; Jian-Ying BAI
Journal of Regional Anatomy and Operative Surgery 2024;33(10):849-854
Objective To construct an artificial intelligence assisted diagnosis model based on deep learning for dynamically recognizing gastric lesions and their locations under gastroscopy in real time,and to evaluate its ability to detect and recognize gastric lesions and their locations.Methods The gastroscopy videos of 104 patients in our hospital was retrospectively analyzed,and the video frames were manually annotated.The annotated picture frames of lesion category were divided into the training set and the validation set according to the ratio of 8∶2,and the annotated picture frames of location category were divided into the training set and the validation set according to the patient sources at the ratio of 8∶2.These sets were utilized for training and validating the respective models.YoloV4 model was used for the training of lesion recognition,and ResNet152 model was used for the training of location recognition.The accuracy,sensitivity,specificity,positive predictive value,negative predictive value and location recognition accuracy of the auxiliary diagnostic model were evaluated.Results A total of 68 351 image frames were annotated,with 54 872 frames used as the training set,including 41 692 frames for lesion categories and 13 180 frames for location categories.The validation set consisted of 13 479 frames,comprising 10 422 frames for lesion categories and 3 057 frames for location categories.The lesion recognition model achieved an overall accuracy of 98.8%,with a sensitivity of 96.6%,specificity of 99.3%,positive predictive value of 96.3%,and negative predictive value of 99.3% in validation set.Meanwhile,the location recognition model demonstrated an top-5 accuracy of 87.1% .Conclusion The artificial intelligence assisted diagnosis model based on deep learning for real-time dynamic recognition of gastric lesions and their locations under gastroscopy has good ability in lesion detection and location recognition,and has great clinical application prospects.
2.Hemichorea associated with non-ketotic hyperglycemia and cerebral blood flow hypoperfusion
Xiaoxiao NIE ; Linlin CHAO ; Mengmeng CAI ; Jingjing SHI ; Jianhua ZHAO
Chinese Journal of Geriatric Heart Brain and Vessel Diseases 2024;26(9):1069-1072
Objective To analyze the clinical and imaging features of hemichorea associated with non-ketotic hyperglycemia(HC-NH)and to explore the perfusion of cerebral blood flow in the patients.Methods A retrospective study was conducted on 23 HC-NH patients diagnosed in Henan Provincial People's Hospital from January 2018 to December 2023.The clinical manifesta-tions,imaging features and prognosis were collected and analyzed,and the correlation with cere-bral blood flow hypoperfusion was investigated.Results The symptoms were all lateral involun-tary movements,of which 4 cases presented only single upper limb(1 case was left upper limb,the other 3 cases were right upper limb),and 19 cases had both upper and lower limbs involved(10 cases were left limb,and 9 cases were right limb).After the onset of the symptoms,the blood glucose level was 19.72±4.72 mmol/L,glycated hemoglobin level was(13.60±3.68)%,but all of patients were negative to urine ketone bodies.Hyperdense lesions in the contralateral basal ganglia region on CT images were observed in 6 cases.Strip or patchy hyperintensity was seen on T1-weighted MR images.All patients had ipsilateral stenosis of the vessels and regional hypoperfu-sion of cerebral blood flow as shown by MR perfusion-weighted imaging.All symptoms were re-lieved after actively controlling blood glucose,improving blood circulation,and symptomatic man-agement.Conclusion HC-NH is quite rare in clinical practice,and its occurrence may be related to cerebral blood flow hypoperfusion triggered by basal nucleus degeneration.
3.Expression of FBXW12 in pancreatic cancer tissues and its effect on the invasion and migration of pancreatic cancer cells
Jinming ZHANG ; Yongfeng XIE ; Jiao NIE ; Lin LU ; Chao DU
China Modern Doctor 2024;62(21):80-85
Objective To investigate the expression of FBXW12 in pancreatic cancer and elucidate its impact on cancer cell migration and invasion.Methods The present study utilized the GEPIA 2 database to analyze the differential expression of FBXW12 between pancreatic cancer tissues and normal tissues.Clinical data from 31 pancreatic cancer patients who underwent radical resection at Linyi People's Hospital from June 2016 to December 2022 were collected.Immunohistochemical staining was conducted to assess FBXW12 expression in both cancerous and adjacent normal tissues obtained during surgery,with subsequent follow-up for survival prognosis.Western blot and polymerase chain reaction(PCR)techniques were employed to determine FBXW12 protein expression levels in pancreatic cancer cell lines.The impact of FBXW12 on cancer cell invasion and migration was evaluated using Transwell cell invasion and scratch test.Results The results from the analysis of the GEPIA 2 database revealed a significant downregulation of FBXW12 mRNA expression in pancreatic cancer tissues compared to normal pancreatic tissues(P<0.05).Immunohistochemical analysis demonstrated a positive expression rate of FBXW12 protein in pancreatic cancer tissues at 75.19%(23/31),whereas adjacent normal tissues exhibited a higher positive expression rate at 93.55%(29/31),indicating a statistically significant difference in FBXW12 expression between pancreatic cancer and adjacent normal tissues(P<0.05).Additionally,the expression level of FBXW12 in pancreatic cancer tissues was found to be closely associated with lymph node metastasis(P<0.05),patients with low expression of FBXW12 have a worse prognosis(P<0.05).Furthermore,transfection with FBXW12 overexpression plasmid resulted in a significant decrease in the invasion and migration abilities of pancreatic cancer cells.Conclusion FBXW12 is low expressed in pancreatic cancer and is associated with the occurrence and development of pancreatic cancer.
4.Application of miniprobe endoscopic ultrasound in endoscopic surgery of small-diameter and low-grade rectum neuroendocrine neoplasm
Jian-Jun LI ; Chao-Qiang FAN ; Xin YANG ; Xue PENG ; Hao LIN ; Xu-Biao NIE ; Shi-Ming YANG ; Qiu-Jian QIAO ; Jian-Ying BAI
Journal of Regional Anatomy and Operative Surgery 2024;33(1):59-62
Objective To evaluate the value of miniprobe endoscopic ultrasound(EUS)in guiding endoscopic treatment of small-diameter(maximum diameter less than 1 cm)and low-grade(G1 grade)rectum neuroendocrine neoplasm(R-NEN),and to provide evidence and clues for its clinical application and further research.Methods The clinical data of 85 cases of low-grade(G1 grade)R-NEN with a maximum diameter of less than 1 cm who underwent endoscopic treatment in our center from January 2014 to December 2020 were retrospectively analyzed.The patients were divided into the EUS group(37 cases)and control group(48 cases)according to whether EUS was performed before endoscopic treatment.The positive rate of incision margin,the incidence of complications,the recurrence rate,the hospital stay,the cost of hospitalization and endoscopic therapy were compared between the two groups.Results The positive rate of incision margin in the EUS group was significantly lower than that in control group(P<0.05).There was no significant difference in the incidence of complications,tumor recurrence rate,hospital stay or hospital costs between the two groups(P>0.05).There was statistically significant difference in the endoscopic therapy between the two groups(P<0.05).Conclusion Evaluating the lesion depth of small-diameter and low-grade(G1 grade)R-NEN before surgery by miniprobe EUS and selecting endoscopic surgery according to its results of can significantly reduce the residual risk of resection margin tumors.
5.Effects of diosgenin on autophagy of human osteosarcoma cells
Chao NIE ; Hua-Ming HUANG ; Bao-Quan HOU ; Jie ZHOU ; Lei ZHANG
Chinese Traditional Patent Medicine 2024;46(1):100-106
AIM To investigate the effects of diosgenin on autophagy of human osteosarcoma cells.METHODS Human osteosarcoma MG63 and U2OS cells with or without exposure to diosgenin had their proliferation detected by MTT assay,their ultrastructure observed by transmission electron microscopy,their expression of autophagy protein Beclin1 observed by immunofluorescence staining,and their expressions of autophagy molecular markers LC3,Beclin1 and PI3K/Akt/mTOR signaling pathway related proteins detected by Western blot.The MG63 and U2OS cells cotreated with diosgenin and PI3K pathway inhibitor LY294002 had the expression of Beclin1 mRNA detected by RT-qPCR.The MG63 and U2OS cells cotreated with autophagy inhibitor 3-methyladenine(3-MA)had their inhibition rate of proliferation detected by MTT assay,their expression of cleaved-caspase3 protein detected by Western blot,and their expression of caspase3 mRNA detected by RT-qPCR.RESULTS Upon osteosarcoma MG63 and U2OS cells,diosgenin inhibited their proliferation,promoted the generation of autophagosomes,increased the protein expression of LC3 Ⅱ and Beclin1(P<0.05,P<0.01),reduced the protein expression of LC3 I(P<0.01),and inhibited the protein phosphorylation level of PI3K/Akt/mTOR pathway(P<0.05,P<0.01),whose effects were offset by the intervention with autophagy inhibitors in terms of the reduced proliferation inhibition and down-regulated expressions of caspase3 mRNA and cleaved-caspase3 protein(P<0.01).CONCLUSION Diosgenin can inhibit the proliferation of osteosarcoma cells and induce their autophagy leading to their death and autophagy apoptosis,which may be related to the activation of PI3K/Akt/mTOR signaling pathway and up-regulation of the expression of LC3 Ⅱ and Beclin1 proteins.
6.Development of a High-throughput Sequencing Platform for Detection of Viral Encephalitis Pathogens Based on Amplicon Sequencing
Li Ya ZHANG ; Zhe Wen SU ; Chen Rui WANG ; Yan LI ; Feng Jun ZHANG ; Hui Sheng LIU ; He Dan HU ; Xiao Chong XU ; Yu Jia YIN ; Kai Qi YIN ; Ying HE ; Fan LI ; Hong Shi FU ; Kai NIE ; Dong Guo LIANG ; Yong TAO ; Tao Song XU ; Feng Chao MA ; Yu Huan WANG
Biomedical and Environmental Sciences 2024;37(3):294-302
Objective Viral encephalitis is an infectious disease severely affecting human health.It is caused by a wide variety of viral pathogens,including herpes viruses,flaviviruses,enteroviruses,and other viruses.The laboratory diagnosis of viral encephalitis is a worldwide challenge.Recently,high-throughput sequencing technology has provided new tools for diagnosing central nervous system infections.Thus,In this study,we established a multipathogen detection platform for viral encephalitis based on amplicon sequencing. Methods We designed nine pairs of specific polymerase chain reaction(PCR)primers for the 12 viruses by reviewing the relevant literature.The detection ability of the primers was verified by software simulation and the detection of known positive samples.Amplicon sequencing was used to validate the samples,and consistency was compared with Sanger sequencing. Results The results showed that the target sequences of various pathogens were obtained at a coverage depth level greater than 20×,and the sequence lengths were consistent with the sizes of the predicted amplicons.The sequences were verified using the National Center for Biotechnology Information BLAST,and all results were consistent with the results of Sanger sequencing. Conclusion Amplicon-based high-throughput sequencing technology is feasible as a supplementary method for the pathogenic detection of viral encephalitis.It is also a useful tool for the high-volume screening of clinical samples.
7.Clinical and imaging characteristics of central pontine myelinolysis without hyponatremia
Linlin CHAO ; Mengmeng CAI ; Jingjing SHI ; Xiaoxiao NIE ; Jianhua ZHAO
Chinese Journal of Neuromedicine 2023;22(12):1260-1263
Objective:To investigate the clinical and imaging characteristics of central pontine myelinolysis (CPM) without hyponatremia and explore its pathogenesis.Methods:A retrospective analysis was performed. Six CPM patients without hyponatremia, admitted to Department of Neurology, He'nan Provincial People's Hospital from March 2021 to March 2023 were chosen. Demographic information, causes, medical history, clinical presentations, and MRI features at onset, and 1 and 3 months after onset were analyzed. The prognoses were evaluated by modified Rankin Scale (mRS) scores 3 months after onset: mRS scores≤2 was classified as good prognosis, and mRS scores>2 as poor prognosis.Results:In these 6 CPM patients without hyponatremia, 4 were males and 2 females; 4 patients had dizziness, 3 headache, 4 limb weakness, 2 cognitive decline, and 2 slow reaction. Four CPM patients had a history of hypertension, 5 had a history of diabetes, and 1 had a history of alcoholic cirrhosis. Hormone therapy, nutritional support and symptomatic treatment were given; 5 patients had obvious improvement, and 1 had poor prognosis 3 months after onset. MRI showed asymmetrical abnormal signal in the basal pons and bilateral brachium pontis, with T1WI hypointensity, T2WI hyperintensity, T2-FLAIR hyperintensity, DWI hyperintensity and clear boundary, and without obvious mass effect or enhancement. DWI sequence enjoyed good diagnostic sensitivity in early stage of CPM: high signal changes could occur within 24 h of clinical symptoms, and isointensity 3 months after onset.Conclusion:Causes of CPM without hyponatremia are mostly hypokalemia, diabetes, malnutrition, and chronic alcoholism; its characteristic MRI manifestations are "pig nose sign", "bat wing sign" and "trident sign".
8.Risk factors and prediction model of perioperative esophagogastric anastomotic leakage after esophageal cancer surgery
Hongxin NIE ; Sihao YANG ; Honggang LIU ; Gaoping CAI ; Dong CHAO ; Hui MENG
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2023;30(04):586-592
Objective To analyze the risk factors for esophagogastric anastomotic leakage (EGAL) after esophageal cancer surgery, and to establish a risk prediction model for early prevention and treatment. Methods Clinical data of patients undergoing esophagectomy in our hospital from January 2013 to October 2020 were retrospectively analyzed. The independent risk factors for postoperative EGAL were analyzed by univariate and multivariate logistic regression analyses, and a clinical nomogram prediction model was established. According to whether EGAL occurred after operation, the patients were divided into an anastomotic fistula group and a non-anastomotic fistula group. Results A total of 303 patiens were enrolled, including 267 males and 36 females with a mean age of 62.30±7.36 years. The incidence rate of postoperative EGAL was 15.2% (46/303). The multivariate logistic regression analysis showed that high blood pressure, chronic bronchitis, peptic ulcer, operation way, the number of lymph node dissected, anastomotic way, the number of intraoperative chest drainage tube, tumor location, no-supplementing albumin in the first three days after operation, postoperative pulmonary infection, postoperative use of bronchoscope were the independent risk factors for EGAL after esophageal cancer surgery (P<0.05). A prognostic nomogram model was established based on these factors with the area under the receiver operating characteristic curve of 0.954 (95%CI 0.924-0.975), indicating a high predictive value. Conclusion The clinical prediction model based on 11 perioperative risk factors in the study has a good evaluation efficacy and can promote the early detection, diagnosis and treatment of EGAL.
9.Capacity of decision-making under ambiguity in adult patients with moyamoya disease
Pan WU ; Jiajia NIE ; Yuxing HAO ; Chao LI ; Kai WANG ; Ling WEI
Acta Universitatis Medicinalis Anhui 2023;58(12):2124-2128
Objective To explore whether the capacity of decision-making under ambiguity in adult patients with moyamoya disease(MMD)is impaired and its association with executive function.Methods Neuropsychological scales were used to measure the executive function of forty-one adult patients with MMD and forty-one healthy con-trols(HC),The Iowa Gambling Task(IGT)was utilized to study the capacity of decision-making under ambiguity of the two groups.Correlation analysis was performed between the two types of data.Results ① The total number of favorable choices in the MMD group was significantly lower than that in the HC group,with a statistically signifi-cant difference(Z=-2.782,P<0.01),and the total number of unfavorable choices was significantly higher than that of the controls,with a statistically significant difference(Z=-2.782,P<0.01).The results of ANOVA showed that the net residual value of Block4 and Block5 in MMD group was lower than that of Block4 and Block5 in HC group,respectively,and the differences were statistically significant(t=-3.433,P=0.001;t=-5.271,P<0.001),the net residual value of Block 1 was higher than that of Block 1 in HC group,the difference was statisti-cally significant(t=2.715,P=0.008).② Spearman correlation analysis revealed no association between the net score of IGT,the total number of favorable choices,the total number of unfavorable choices,and executive func-tion.Conclusion The capacity of decision-making under ambiguity in adult patients with MMD is impaired possi-bly,which is not correlated with executive cognition.
10.Serum exosome-mediated lncRNA HOTTIP regulates cisplatin resistance in gastric cancer cells through the miR-138-5p/TJP1 axis
Mingyang HAN ; Junwei BAI ; Jiewei NIE ; Yuanyuan LI ; Chao ZHANG
Chinese Journal of Digestive Surgery 2023;22(12):1467-1475
Objective:To investigate the effects of serum exosomes of patients with gastric cancer on cisplatin resistance, clonal formation, migration, invasion and apoptosis of the AGS gastric cancer cells, and the corresponding molecular mechanisms.Methods:The experimental study was conducted. The exosomes of patients with gastric cancer was separated from their serum, and the expression of lncRNA HOTTIP was analyzed using the quantitative real time polymerase chain reaction (qRT-PCR). Normal gastric epithelial cell line GES1, gastric cancer cell line AGS and human embryonic kidney cell 293T were cultured in vitro. AGS cells were incubated with exosomes (Exo),with phos-phate buffered saline (PBS) treatment as control, and transfected with si-NC or si-HOTTIP-3, named as Exo group, PBS group, si-NC+Exo group, and si-HOTTIP-3+Exo group. The AGS cells were trans-fected with si-NC, si-HOTTIP-1, si-HOTTIP-2, si-HOTTIP-3, oe-HOTTIP, vector, oe-HOTTIP+miR-138-5p mimic, oe-HOTTIP+mimic NC, miR-138-5p inhibitor, inhibitor NC, miR-138-5p inhibitor+si-TJP1 and miR-138-5p inhibitor+si-NC. They were recorded as si-NC group, si-HOTTIP-1 group, si-HOTTIP-2 group, si-HOTTIP-3 group, oe-HOTTIP group, vector group, oe-HOTTIP+miR-138-5p mimic group, oe-HOTTIP+mimic NC group, miR-138-5p inhibitor group, inhibitor NC group, miR-138-5p inhibitor+si-TJP1 group and miR-138-5p inhibitor+si-NC group. The 293T cells transfected with mimic NC+HOTTIP wt, miR-138-5p mimic+HOTTIP wt, mimic NC+HOTTIP mut, miR-138-5p mimic+HOTTIP mut, mimic NC+TJP1 3'UTR wt, miR-138-5p mimic+TJP1 3'UTR wt, mimic NC+TJP1 3'UTR mut, miR-138-5p mimic+TJP1 3'UTR mut were recorded as the mimic NC+HOTTIP wt group, miR-138-5p mimic+HOTTIP wt group, mimic NC+HOTTIP mut group, miR-138-5p mimic+HOTTIP mut group, mimic NC+TJP1 3'UTR wt group, miR-138-5p mimic+TJP1 3'UTR wt group, mimic NC+TJP1 3'UTR mut group, miR-138-5p mimic+TJP1 3'UTR mut group. The cell counting kit-8 (CCK8) was used to analyze the cisplatin sensitivity of gastric cancer cells. The colony formation experiment was used to analyze the colony formation of gastric cancer cells. The Transwell experiment was used to analyzed cell migration and invasion of gastric cancer cells. The flow cytometry experiment was used to analyze cell apoptosis of gastric cancer cells. The Western bolt assay was used to analyze the expression of exosome marker proteins, including the CD63 and CD81, and the protein of TJP1, the drug-resistance related proteins, including the P-gp and MCL-1. The dual-luciferase assay was used to analyze the targeted relationships among lncRNA HOTTIP, miR-138-5p and TJP1. Observation indicators: (1) expression of lncRNA HOTTIP; (2) resistance of gastric cancer cells to cisplatin regulated by exosome-mediated lncRNA HOTTIP; (3) regulation of cisplatin resistance in gastric cancer cells mediated by miR-138-5p through lncRNA HOTTIP overexpression; (4) targeting of TJP1 gene 3′-untranslated region (UTR) by miR-138-5p; (5) regulation of cisplatin resistance in gastric cancer cells by TJP1 through miR-138-5p inhibition. Measurement data with normal distribution were represented as Mean± SD, and comparison between groups was conducted using the t test. The one-way ANOVA was used for comparison for multiple groups and the Tukey′s test was used for further pairwise compari-son. Count data were described as absolute numbers, and the chi-square test was used for comparison. Correlation analysis was conducted using the Pearson′s test. Results:(1) Expression of lncRNA HOTTIP. The expression of lncRNA HOTTIP in the serum exosome of patients with gastric cancer was higher than that in healthy volunteers, showing a significant difference ( P<0.05). Results of transmi-ssion electron microscopy examination showed that the serum exosomes were circular or oval in shape. Results of Western bolt assay showed the expression of marker proteins of CD63 and CD81 in serum exosomes. (2) Resistance of gastric cancer cells to cisplatin regulated by exosome-mediated lncRNA HOTTIP. Compared with the PBS group, the biochemical half maximal inhibitory concentra-tion (IC50), the number of clone formation, the number of invasive cell, the number of migratory cell, expression of P-gp protein, expression of MCL-1 protein in the Exo group increased, while the cell apoptosis rate decreased, showing significant differences between them ( P<0.05). Compared with the si-NC+Exo group, the IC50, the number of clone formation, the number of invasive cell, the number of migratory cell, expression of P-gp protein, expression of MCL-1 protein in the si-HOTTIP-3+Exo group decreased, while the cell apoptosis rate increased, showing significant differences between them ( P<0.05). (3) Regulation of cisplatin resistance in gastric cancer cells mediated by miR-138-5p through lncRNA HOTTIP overexpression. Compared with the vector group, the IC50, the number of clone formation, the number of invasive cell, the number of migratory cell, expression of P-gp protein, expression of MCL-1 protein in the oe-HOTTIP group increased, while the cell apoptosis rate decreased, showing significant differences between them ( P<0.05). Compared with the oe-HOTTIP+mimic NC group, the IC50, the number of clone formation, the number of invasive cell, the number of migratory cell, expression of P-gp protein, expression of MCL-1 protein in the oe-HOTTIP+miR-138-5p mimic group increased, while the cell apoptosis rate decreased, showing significant differences between them ( P<0.05). (4) Targeting of TJP1 gene 3′-UTR by miR-138-5p. Results of dual-luciferase assay showed that the luciferase activity in 293T cells treatment with mimics of control+vectors of wild type of TJP1 gene 3′-UTR and 293T cells treatment with mimics of miR-138-5p+vectors of wild type of TJP1 gene 3′-UTR was 1.00±0.09 and 0.21±0.03, respectively, showing a significant difference between them ( t=15.02, P<0.05). (5) Regulation of cisplatin resistance in gastric cancer cells by TJP1 through miR-138-5p inhibition. Compared with the inhibitor group, the IC50, the number of clone formation, the number of invasive cell, the number of migratory cell, expression of P-gp protein, expression of MCL-1 protein in the miR-138-5p inhibitor group increased, while the cell apoptosis rate decreased, showing significant differences between them ( P<0.05). Compared with the miR-138-5p inhibitor+si-NC group, the IC50, the number of clone formation, the number of invasive cell, the number of migratory cell, expression of P-gp protein, expression of MCL-1 protein in the miR-138-5p inhibitor+si-TJP1 group decreased, while the cell apoptosis rate increased, showing significant differences between them ( P<0.05). Conclusion:Serum exosomes-mediated lncRNA HOTTIP can promote cisplatin resistance, clonal formation, migration, invasion and apoptosis of gastric cancer cells and inhibit cell apoptosis of gastric cancer cells through regulating the expression of miR-138-5p/TJP1.

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