1.Safety of high-carbohydrate fluid diet 2 h versus overnight fasting before non-emergency endoscopic retrograde cholangiopancreatography: A single-blind, multicenter, randomized controlled trial
Wenbo MENG ; W. Joseph LEUNG ; Zhenyu WANG ; Qiyong LI ; Leida ZHANG ; Kai ZHANG ; Xuefeng WANG ; Meng WANG ; Qi WANG ; Yingmei SHAO ; Jijun ZHANG ; Ping YUE ; Lei ZHANG ; Kexiang ZHU ; Xiaoliang ZHU ; Hui ZHANG ; Senlin HOU ; Kailin CAI ; Hao SUN ; Ping XUE ; Wei LIU ; Haiping WANG ; Li ZHANG ; Songming DING ; Zhiqing YANG ; Ming ZHANG ; Hao WENG ; Qingyuan WU ; Bendong CHEN ; Tiemin JIANG ; Yingkai WANG ; Lichao ZHANG ; Ke WU ; Xue YANG ; Zilong WEN ; Chun LIU ; Long MIAO ; Zhengfeng WANG ; Jiajia LI ; Xiaowen YAN ; Fangzhao WANG ; Lingen ZHANG ; Mingzhen BAI ; Ningning MI ; Xianzhuo ZHANG ; Wence ZHOU ; Jinqiu YUAN ; Azumi SUZUKI ; Kiyohito TANAKA ; Jiankang LIU ; Ula NUR ; Elisabete WEIDERPASS ; Xun LI
Chinese Medical Journal 2024;137(12):1437-1446
		                        		
		                        			
		                        			Background::Although overnight fasting is recommended prior to endoscopic retrograde cholangiopancreatography (ERCP), the benefits and safety of high-carbohydrate fluid diet (CFD) intake 2 h before ERCP remain unclear. This study aimed to analyze whether high-CFD intake 2 h before ERCP can be safe and accelerate patients’ recovery.Methods::This prospective, multicenter, randomized controlled trial involved 15 tertiary ERCP centers. A total of 1330 patients were randomized into CFD group ( n = 665) and fasting group ( n = 665). The CFD group received 400 mL of maltodextrin orally 2 h before ERCP, while the control group abstained from food/water overnight (>6 h) before ERCP. All ERCP procedures were performed using deep sedation with intravenous propofol. The investigators were blinded but not the patients. The primary outcomes included postoperative fatigue and abdominal pain score, and the secondary outcomes included complications and changes in metabolic indicators. The outcomes were analyzed according to a modified intention-to-treat principle. Results::The post-ERCP fatigue scores were significantly lower at 4 h (4.1 ± 2.6 vs. 4.8 ± 2.8, t = 4.23, P <0.001) and 20 h (2.4 ± 2.1 vs. 3.4 ± 2.4, t= 7.94, P <0.001) in the CFD group, with least-squares mean differences of 0.48 (95% confidence interval [CI]: 0.26–0.71, P <0.001) and 0.76 (95% CI: 0.57–0.95, P <0.001), respectively. The 4-h pain scores (2.1 ± 1.7 vs. 2.2 ± 1.7, t = 2.60, P = 0.009, with a least-squares mean difference of 0.21 [95% CI: 0.05–0.37]) and positive urine ketone levels (7.7% [39/509] vs. 15.4% [82/533], χ2 = 15.13, P <0.001) were lower in the CFD group. The CFD group had significantly less cholangitis (2.1% [13/634] vs. 4.0% [26/658], χ2 = 3.99, P = 0.046) but not pancreatitis (5.5% [35/634] vs. 6.5% [43/658], χ2 = 0.59, P = 0.444). Subgroup analysis revealed that CFD reduced the incidence of complications in patients with native papilla (odds ratio [OR]: 0.61, 95% CI: 0.39–0.95, P = 0.028) in the multivariable models. Conclusion::Ingesting 400 mL of CFD 2 h before ERCP is safe, with a reduction in post-ERCP fatigue, abdominal pain, and cholangitis during recovery.Trail Registration::ClinicalTrials.gov, No. NCT03075280.
		                        		
		                        		
		                        		
		                        	
2.Clinical characteristics of choledocholithiasis combined with periampullary diverticulum and influencing factor analysis for difficult cannulation of endoscopic retrograde cholangiopan-creatography: a report of 1 920 cases
Ping YUE ; Zhenyu WANG ; Leida ZHANG ; Hao SUN ; Ping XUE ; Wei LIU ; Qi WANG ; Jijun ZHANG ; Xuefeng WANG ; Meng WANG ; Yingmei SHAO ; Kailin CAI ; Senlin HOU ; Kai ZHANG ; Qiyong LI ; Lei ZHANG ; Kexiang ZHU ; Haiping WANG ; Ming ZHANG ; Xiangyu SUN ; Zhiqing YANG ; Jie TAO ; Zilong WEN ; Qunwei WANG ; Bendong CHEN ; Yingkai WANG ; Mingning ZHAO ; Ruoyan ZHANG ; Tiemin JIANG ; Ke LIU ; Lichao ZHANG ; Kangjie CHEN ; Xiaoliang ZHU ; Hui ZHANG ; Long MIAO ; Zhengfeng WANG ; Jiajia LI ; Xiaowen YAN ; Ling'en ZHANG ; Fangzhao WANG ; Wence ZHOU ; Wenbo MENG ; Xun LI
Chinese Journal of Digestive Surgery 2023;22(1):113-121
		                        		
		                        			
		                        			Objective:To investigate the clinical characteristics of choledocholithiasis com-bined with periampullary diverticulum and influencing factor for difficult cannulation of endoscopic retrograde cholangiopancreatography (ERCP).Methods:The retrospective case-control study was conducted. The clinical data of 1 920 patients who underwent ERCP for choledocholithiasis in 15 medical centers, including the First Hospital of Lanzhou University, et al, from July 2015 to December 2017 were collected. There were 915 males and 1 005 females, aged (63±16)years. Of 1 920 patients, there were 228 cases with periampullary diverticulum and 1 692 cases without periampullary diverticulum. Observation indicators: (1) clinical characteristics of patients with choledocholithiasis; (2) intraoperative and postoperative situations of patients undergoing ERCP for choledocholithiasis; (3) influencing factor analysis for difficult cannulation in patients undergoing ERCP for choledocholithiasis. Measurement data with normal distribution were represented as Mean±SD, and comparison between groups was conducted using the independent sample t test. Measurement data with skewed distribution were represented as M(range) or M( Q1, Q3), and com-parison between groups was conducted using the Wilcoxon rank sum test. Count data were described as absolute numbers or percentages, and comparison between groups was conducted using the chi-square test or Fisher exact probability. The Logistic regression model was used for univariate and multivariate analyses. Results:(1) Clinical characteristics of patients with choledocholithiasis. Age, body mass index, cases with complications as chronic obstructive pulmonary disease, diameter of common bile duct, cases with diameter of common bile duct as <8 mm, 8?12 mm, >12 mm, diameter of stone, cases with number of stones as single and multiple were (69±12)years, (23.3±3.0)kg/m 2, 16, (14±4)mm, 11, 95, 122, (12±4)mm, 89, 139 in patients with choledocholithiasis combined with periampullary diverticulum, versus (62±16)years, (23.8±2.8)kg/m 2, 67, (12±4)mm, 159, 892, 641, (10±4)mm, 817, 875 in patients with choledocholithiasis not combined with periampullary diver-ticulum, showing significant differences in the above indicators between the two groups ( t=?7.55, 2.45, χ2=4.54, t=?4.92, Z=4.66, t=?7.31, χ2=6.90, P<0.05). (2) Intraoperative and postoperative situations of patients undergoing ERCP for choledocholithiasis. The balloon expansion diameter, cases with intraoperative bleeding, cases with hemorrhage management of submucosal injection, hemostatic clip, spray hemostasis, electrocoagulation hemostasis and other treatment, cases with endoscopic plastic stent placement, cases with endoscopic nasal bile duct drainage, cases with mechanical lithotripsy, cases with stone complete clearing, cases with difficult cannulation, cases with delayed intubation, cases undergoing >5 times of cannulation attempts, cannulation time, X-ray exposure time, operation time were 10.0(range, 8.5?12.0)mm, 56, 6, 5, 43, 1, 1, 52, 177, 67, 201, 74, 38, 74, (7.4±3.1)minutes, (6±3)minutes, (46±19)minutes in patients with choledocholithiasis combined with periampullary diverticulum, versus 9.0(range, 8.0?11.0)mm, 243, 35, 14, 109, 73, 12, 230, 1 457, 167, 1 565, 395, 171, 395, (6.6±2.9)minutes, (6±5)minutes, (41±17)minutes in patients with choledocholithiasis not combined with periampullary diverticulum, showing significant differences in the above indicators between the two groups ( Z=6.31, χ2=15.90, 26.02, 13.61, 11.40, 71.51, 5.12, 9.04, 8.92, 9.04, t=?3.89, 2.67, ?3.61, P<0.05). (3) Influencing factor analysis for difficult cannulation in patients undergoing ERCP for choledocholithiasis. Results of multivariate analysis showed total bilirubin >30 umol/L, number of stones >1, combined with periampullary diverticulum were indepen-dent risk factors for difficult cannulation in patients with periampullary diverticulum who underwent ERCP for choledocholithiasis ( odds ratio=1.31, 1.48, 1.44, 95% confidence interval as 1.06?1.61, 1.20?1.84, 1.06?1.95, P<0.05). Results of further analysis showed that, of 1 920 patients undergoing ERCP for choledocholithiasis, the incidence of postoperative pancreatitis was 17.271%(81/469) and 8.132%(118/1 451) in the 469 cases with difficult cannulation and 1 451 cases without difficult cannula-tion, respectively, showing a significant difference between them ( χ2=31.86, P<0.05). In the 1 692 patients with choledocholithiasis not combined with periampullary diverticulum, the incidence of postopera-tive pancreatitis was 17.722%(70/395) and 8.250%(107/1 297) in 395 cases with difficult cannula-tion and 1 297 cases without difficult cannulation, respectively, showing a significant difference between them ( χ2=29.00, P<0.05). In the 228 patients with choledocholithiasis combined with peri-ampullary diverticulum, the incidence of postoperative pancreatitis was 14.865%(11/74) and 7.143%(11/154) in 74 cases with difficult cannulation and 154 cases without difficult cannulation, respectively, showing no significant difference between them ( χ2=3.42, P>0.05). Conclusions:Compared with patients with choledocholithiasis not combined with periampullary divertioulum, periampullary divertioulum often occurs in choledocholithiasis patients of elderly and low body mass index. The proportion of chronic obstructive pulmonary disease is high in choledocholithiasis patients with periampullary diverticulum, and the diameter of stone is large, the number of stone is more in these patients. Combined with periampullary diverticulum will increase the difficult of cannulation and the ratio of patient with mechanical lithotripsy, and reduce the ratio of patient with stone complete clearing without increasing postoperative complications of choledocholithiasis patients undergoing ERCP. Total bilirubin >30 μmol/L, number of stones >1, combined with periampullary diverticulum are independent risk factors for difficult cannulation in patients of periampullary diverticulum who underwent ERCP for choledocholithiasis.
		                        		
		                        		
		                        		
		                        	
3.Effect of pulmonary rehabilitation training based on GOLD classification in patients with acute exacerbation of chronic obstructive pulmonary disease
Ronghui JIN ; Wei FANG ; Jianjun ZHAO ; Yingmei QI
Chinese Journal of Modern Nursing 2022;28(35):4915-4920
		                        		
		                        			
		                        			Objective:To explore the effect of pulmonary rehabilitation training based on Chronic Obstructive Pulmonary Disease (GOLD) classification on self-efficacy and rehabilitation effect in patients with acute exacerbation of chronic obstructive pulmonary disease.Methods:From May 2019 to April 2021, a total of 85 patients with acute exacerbation of chronic obstructive pulmonary disease who were admitted to China-Japan Union Hospital of Jilin University were selected and divided into the observation group (43 cases) and the control group (42 cases) using the random number table method. The control group received routine pulmonary rehabilitation training, and the observation group conducted pulmonary rehabilitation training based on GOLD classification. Two weeks after intervention, the self-efficacy, pulmonary function, 6-Minute Walking Test (6MWT), quality of life, and the condition of adverse reactions were compared between the two groups.Results:Two weeks after intervention, the scores of dyspnea management, emotion, physical activity, safety behavior, and total score of self-efficacy of the observation group were higher than those of the control group, and the differences were statistically significant ( P<0.05). The forced exhalation volume in one second (FEV1), forced vital capacity (FVC), FEV1/FVC and 6MWT of the observation group were higher than those of the control group, and the differences were statistically significant ( P<0.05). The scores of respiratory symptoms, activity limitation, disease impact on life and total score of St. George's Respiratory Questionnaire (SGRQ) of the observation group were lower than those of the control group, and the differences were statistically significant ( P<0.05). There were no serious adverse reactions in two groups. Conclusions:Pulmonary rehabilitation training based on GOLD classification can help patients' promote the recovery of lung function, improve their self-efficacy and the quality of life, which has good security.
		                        		
		                        		
		                        		
		                        	
4.Expression and clinical significance of the NK cell-activating receptor NKG2D and its ligand in liver tissue of patients with primary biliary cholangitis
Haiyan YU ; Jinxiang WEI ; Haiyan FU ; Yina YANG ; Rongfang TU ; Yingmei TANG
Journal of Clinical Hepatology 2020;36(11):2456-2461
		                        		
		                        			
		                        			 ObjectiveTo investigate the association of the expression of the NK cell-activating receptor NKG2D, its ligand major histocompatibility complex class I chain-related gene A (MICA), and related cytokines [interferon-γ (IFN-γ), interleukin-10 (IL-10), and interleukin-15 (IL-15)] with intrahepatic inflammation in primary biliary cholangitis (PBC). MethodsLiver biopsy specimens were collected from 30 patients with PBC (PBC group), 15 patients with chronic hepatitis B (CHB group), and 10 patients with nonalcoholic fatty liver disease (NAFLD group), who were hospitalized in The Second Affiliated Hospital of Kunming Medical University from August 2014 to June 2015. The degree of liver inflammation (G) and fibrosis degree (S) of the liver specimens were determined, and immunohistochemistry was used to measure the expression of NKG2D, MICA, IFN-γ, IL-10, and IL-15 in liver tissue (the scores were determined based on the number of cells stained and the degree of staining to evaluate the expression of each marker). A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the t-test was used for comparison between two groups; a Spearman correlation analysis was used to investigate correlation. ResultsIn the PBC group, the expression of NKG2D increased with the degree of inflammation, and the patients with G3-4 inflammation had significantly higher expression than those with G1-2 inflammation (G1 vs G2 vs G3 vs G4: 1.4±0.05 vs 1.56±0.05 vs 1.86±0.11 vs 2.60±0.17, F=150.8, P<0.05); the expression of NKG2D decreased with fibrosis degree (S3 vs S4: 2.30±0.17 vs 1.56±0.05, t=-1.52, P<0.05). In the PBC group, there was no significant difference in MICA between G3 and G4 (0.11±0.01 vs 0.20±0.03, t=-2.20, P>0.05) and between S3 and S4 (0.12±0.02 vs 0.18±0.03, t=-2.64, P>0.05). In the PBC group, there was a significant difference in the expression of IL-15 between the patients with different degrees of inflammation (G1 vs G2 vs G3 vs G4: 0.70±0.10 vs 1.50±0.10 vs 1.93±0.11 vs 2.60±0.17, F=251.3, P<0.05), while there was no significant difference between the patients with different fibrosis degrees (S3 vs S4: 2.00±0.05 vs 2.40±0.30, t=-1.62, P>0.05). In the CHB group, there was a significant difference in the expression of IL-15 between the patients with different degrees of inflammation (G1 vs G2 vs G3: 0.73±0.15 vs 1.96±0.15 vs 2.50±0.17, F=150, P<0.05) and between the patients with different fibrosis degrees (S1 vs S2 vs S3: 0.70±0.10 vs 21.96±0.15 vs 2.50±0.17, F=158.7, P<0.05). In the PBC group, the expression of IL-10 was only observed in the patients with G1 inflammation (0.16±0.01), and in the CHB group, the expression of IL-10 was observed in the patients with G1 and G2 inflammation, with no significant difference (G1 vs G2: 0.19±0.01 vs 0.13±0.01, t=-1.522, P>0.05). In the patients with PBC, the expression of IL-15 in liver tissue was positively correlated with the levels of alkaline phosphatase (ALP) and gamma-glutamyl transpeptidase (GGT) (r=0.241 and 0.407, P=0.014 and 0.045). ConclusionThe NK cell-activating receptor NKG2D affects the degree of intrahepatic inflammation in PBC, and the NKG2D ligand MICA is expressed in the advanced stage of PBC and can downregulate NKG2D. The expression of IL-15 increases with the degree of inflammation in PBC and is positively correlated with the levels of ALP and GGT, suggesting that the activation of NK cells and abnormal secretion of cytokines are involved in the development and progression of PBC and IL-15 may be used as an auxiliary index for the diagnosis of PBC. 
		                        		
		                        		
		                        		
		                        	
5.Application of perioperative nutrition management strategy in patients with hepatic cystic echinococcosis
Zhi LU ; Jiezheng HU ; Huanhuan WEI ; Yingmei SHAO ; Meixin WANG
Chinese Journal of Practical Nursing 2020;36(9):641-646
		                        		
		                        			
		                        			Objective:To explore the application of perioperative nutrition management strategy in patients with hepatic cystic echinococcosis.Methods:From October 2017 to February 2019, 53 patients with hepatic cystic echinococcosis who met the inclusion criteria were selected as the control group. Another 53 patients from June 2018 to January 2019 were collected as the experimental group, the nutritional index, liver function, complication rate and postoperative recovery were compared between the two groups.Results:The levels of retinol-binding protein, prealbumin and transferrin were (24.32 ± 3.76) μg/L, (167.00 ± 24.12) mg/L, (2.08 ± 0.43) μg/L on the day of admission in the experimental group, and one day before the operation were (27.78± 4.98) μg/L, (245.00 ± 22.02) mg/L,(2.47 ± 0.54)μ g/L, there was no significant difference ( t=0.576-3.552, P < 0.05). The incidence of infection, biliary leakage and hemorrhage in the experimental group were 0, 1.89% (1/53) and 0 respectively, which were lower than 9.43% (5/53), 13.21% (7/53), 11.32% (6/53) in the control group ( χ2 value was 4.867, P < 0.05). The aspartate aminotransferase and alanine aminotransferase of the experimental group on the 7th day after operation were (51.50 ± 6.30), (29.54 ± 2.03) U/L, which were significantly different from (69.53 ± 7.07), (43.72±3.67) U/L of the control group ( t value was 2.032, 2.015, P<0.05). Anal exhaust, defecation time, hospitalization time and expenses in the experimental group were (15.89±8.34) h, (49.12±10.56) h, (8.69 ± 1.69) d, (2.84±1.37) thousand yuan, which were significantly different from (34.13±7.13) h, (63.45±11.03) h, (11.51±4.18) d, (3.76±1.53) thousand yuan in the control group. There was statistical significance ( t values were 3.372-12.592, P<0.05). Conclusions:The application of perioperative nutrition management strategy in patients with hepatic cystic echinococcosis can improve the nutritional status of patients during perioperative period, promote the recovery of liver function and reduce the occurrence of postoperative complications.
		                        		
		                        		
		                        		
		                        	
6.Recommendations on management of gynecological malignancies during the COVID-19 pandemic: perspectives from Chinese gynecological oncologists
Yingmei WANG ; Shiqian ZHANG ; Lihui WEI ; Zhongqiu LIN ; Xinyu WANG ; Jianliu WANG ; Keqin HUA ; Manhua CUI ; Jiandong WANG ; Shixuan WANG ; Wen DI ; Yudong WANG ; Ruifang AN ; Mingrong XI ; Ruixia GUO ; Qi ZHOU ; Xing XIE ; Fengxia XUE
Journal of Gynecologic Oncology 2020;31(4):e68-
		                        		
		                        			
		                        			 The outbreak of coronavirus disease 2019 (COVID-19) caused by the severe acute respiratory syndrome coronavirus 2 has rapidly spread globally. Cancer patients are at a higher risk of being infected with the coronavirus and are more likely to develop severe complications, as compared to the general population. The increasing spread of COVID-19 presents challenges for the clinical care of patients with gynecological malignancies. Concerted efforts should be put into managing gynecological malignancies in an orderly manner by strictly implementing the measures that are specifically developed for controlling the spread of COVID-19. We have drafted Recommendations on Management of Gynecological Malignancies during the COVID-19 Pandemic based on our experience on controlling COVID-19 pandemic in China. We recommend that patients with gynecological malignancies should be managed in hierarchical and individualized manners in combination with local conditions related to COVID-19. Medical care decision should be balanced between controlling COVID-19 pandemic spread and timely diagnosis and treatment for gynecologic oncology patients. 
		                        		
		                        		
		                        		
		                        	
7. Downregulation of Herg1 suppresses osteosarcoma proliferation and invasion by targeting Hippo signaling pathway
Zhida CHEN ; Wenbin YE ; Wenrong ZENG ; Chao SONG ; Wei SONG ; Yingmei ZHANG ; Jin WU
Chinese Journal of Oncology 2019;41(5):338-345
		                        		
		                        			 Objective:
		                        			To detect the effect and regulatory mechanism of human ether à go-go related gene 1 (Herg 1) knockdown on the proliferation and invasion of osteosarcoma (OS).
		                        		
		                        			Methods:
		                        			We constructed a recombinant adenovirus vector (Ad5-Herg1-shRNA) expressing short hair RNA (shRNA) against Herg1 and tested the knockdown efficiency. Then, the effects of Herg 1 knockdown on the proliferation, growth and invasion of osteosarcoma were measured by using cell counting kit-8 (CCK-8), wound healing assay, Transwell assay and xenograft model of nude mice, respectively. Tandem affinity purification, mass spectrometry and dual luciferase reporter assay were used to find out the molecules interacted with Herg1. Western blot was used to detect the expressions of large tumor suppressor gene (LATS1), p-LATS1, Yes-associated protein (YAP) and p-YAP in cells after infection of Ad5-Herg1-shRNA.
		                        		
		                        			Results:
		                        			Compared to Ad5-control-shRNA, Ad5-Herg1-shRNA dramatically inhibited the expression of Herg1 in OS cells. The result of CCK8 array demonstrated that 143B cell vitalities of Ad5-Herg1-shRNA1 and Ad5-Herg1-shRNA2 group were (65.47±3.90)% and (79.90±1.52)%, significantly lower than (100.00±6.14)% of Ad5-control-shRNA group. Meanwhile, U2OS cell vitality of Ad5-Herg1-shRNA1 and Ad5-Herg1-shRNA2 group were (69.69±1.36)% and (76.72±2.75)%, significantly lower than (100.00±3.01)% of Ad5-control-shRNA group (all 
		                        		
		                        	
8.Downregulation of Herg1 suppresses osteosarcoma proliferation and invasion by targeting Hippo signaling pathway
Zhida CHEN ; Wenbin YE ; Wenrong ZENG ; Chao SONG ; Wei SONG ; Yingmei ZHANG ; Jin WU
Chinese Journal of Oncology 2019;41(5):338-345
		                        		
		                        			
		                        			Objective To detect the effect and regulatory mechanism of human ether à go?go related gene 1 ( Herg 1) knockdown on the proliferation and invasion of osteosarcoma ( OS). Methods We constructed a recombinant adenovirus vector ( Ad5?Herg1?shRNA) expressing short hair RNA ( shRNA) against Herg1 and tested the knockdown efficiency. Then, the effects of Herg 1 knockdown on the proliferation, growth and invasion of osteosarcoma were measured by using cell counting kit?8 (CCK?8), wound healing assay, Transwell assay and xenograft model of nude mice, respectively. Tandem affinity purification, mass spectrometry and dual luciferase reporter assay were used to find out the molecules interacted with Herg1. Western blot was used to detect the expressions of large tumor suppressor gene (LATS1), p?LATS1, Yes?associated protein ( YAP ) and p?YAP in cells after infection of Ad5?Herg1?shRNA. Results Compared to Ad5?control?shRNA, Ad5?Herg1?shRNA dramatically inhibited the expression of Herg1 in OS cells. The result of CCK8 array demonstrated that 143B cell vitalities of Ad5?Herg1?shRNA1 and Ad5?Herg1?shRNA2 group were ( 65.47 ± 3.90)% and ( 79.90 ± 1.52)%, significantly lower than (100.00±6.14)% of Ad5?control?shRNA group. Meanwhile, U2OS cell vitality of Ad5?Herg1?shRNA1 and Ad5?Herg1?shRNA2 group were (69.69±1.36)% and (76.72±2.75)%, significantly lower than (100.00± 3.01)% of Ad5?control?shRNA group (all P<0.001). The results of wound healing array showed that 143B cell migration rates of Ad5?Herg1?shRNA1 and Ad5?Herg1?shRNA2 group were (33.03± 2.88)% and (36.47±4.16)%, significantly lower than (97.78±2.28)% of Ad5?control?shRNA group. Meanwhile, U2OS cell migration rates of Ad5?Herg1?shRNA1 and Ad5?Herg1?shRNA2 group were ( 68.07 ± 0.90 )% and (73.97±1.25)%, significantly lower than (96.50± 1.12)% of Ad5?control?shRNA group ( all P<0.001). The results of Transwell showed that 143B cell invasion numbers of Ad5?Herg1?shRNA1 and Ad5?Herg1?shRNA2 group were 36.50±12.15 and 44.83±7.62, significantly lower than 195.33±19.68 of Ad5?control?shRNA group. Meanwhile, U2OS cell migration rates of Ad5?Herg1?shRNA1 and Ad5?Herg1?shRNA2 group were 21.83±7.99 and 22.85±7.08, significantly lower than 83.33±12.36 of Ad5?control?shRNA group ( all P<0.001). The results of xenograft model of OS showed that the tumor volume and weight of Ad5?Herg1?shRNA group were significantly smaller than of Ad5?control?shRNA group after 14 days and 5 weeks of inoculation, respectively (P<0.001).Moreover, knockdown of Herg1 inhibited the metastasis of OS cells.In mechanism, Herg1 protein interacted with NF2 protein. Knockdown of Herg1 significantly suppressed the expression levels of LATS1 and YAP protein, and promoted the phosphorylation of LATS1 and YAP in OS cells ( all P<0.001). Conclusion Our findings suggest that Herg1 participates in the proliferation and motility of OS cells and may serve as a potential therapeutic target for osteosarcoma patients.
		                        		
		                        		
		                        		
		                        	
9.Downregulation of Herg1 suppresses osteosarcoma proliferation and invasion by targeting Hippo signaling pathway
Zhida CHEN ; Wenbin YE ; Wenrong ZENG ; Chao SONG ; Wei SONG ; Yingmei ZHANG ; Jin WU
Chinese Journal of Oncology 2019;41(5):338-345
		                        		
		                        			
		                        			Objective To detect the effect and regulatory mechanism of human ether à go?go related gene 1 ( Herg 1) knockdown on the proliferation and invasion of osteosarcoma ( OS). Methods We constructed a recombinant adenovirus vector ( Ad5?Herg1?shRNA) expressing short hair RNA ( shRNA) against Herg1 and tested the knockdown efficiency. Then, the effects of Herg 1 knockdown on the proliferation, growth and invasion of osteosarcoma were measured by using cell counting kit?8 (CCK?8), wound healing assay, Transwell assay and xenograft model of nude mice, respectively. Tandem affinity purification, mass spectrometry and dual luciferase reporter assay were used to find out the molecules interacted with Herg1. Western blot was used to detect the expressions of large tumor suppressor gene (LATS1), p?LATS1, Yes?associated protein ( YAP ) and p?YAP in cells after infection of Ad5?Herg1?shRNA. Results Compared to Ad5?control?shRNA, Ad5?Herg1?shRNA dramatically inhibited the expression of Herg1 in OS cells. The result of CCK8 array demonstrated that 143B cell vitalities of Ad5?Herg1?shRNA1 and Ad5?Herg1?shRNA2 group were ( 65.47 ± 3.90)% and ( 79.90 ± 1.52)%, significantly lower than (100.00±6.14)% of Ad5?control?shRNA group. Meanwhile, U2OS cell vitality of Ad5?Herg1?shRNA1 and Ad5?Herg1?shRNA2 group were (69.69±1.36)% and (76.72±2.75)%, significantly lower than (100.00± 3.01)% of Ad5?control?shRNA group (all P<0.001). The results of wound healing array showed that 143B cell migration rates of Ad5?Herg1?shRNA1 and Ad5?Herg1?shRNA2 group were (33.03± 2.88)% and (36.47±4.16)%, significantly lower than (97.78±2.28)% of Ad5?control?shRNA group. Meanwhile, U2OS cell migration rates of Ad5?Herg1?shRNA1 and Ad5?Herg1?shRNA2 group were ( 68.07 ± 0.90 )% and (73.97±1.25)%, significantly lower than (96.50± 1.12)% of Ad5?control?shRNA group ( all P<0.001). The results of Transwell showed that 143B cell invasion numbers of Ad5?Herg1?shRNA1 and Ad5?Herg1?shRNA2 group were 36.50±12.15 and 44.83±7.62, significantly lower than 195.33±19.68 of Ad5?control?shRNA group. Meanwhile, U2OS cell migration rates of Ad5?Herg1?shRNA1 and Ad5?Herg1?shRNA2 group were 21.83±7.99 and 22.85±7.08, significantly lower than 83.33±12.36 of Ad5?control?shRNA group ( all P<0.001). The results of xenograft model of OS showed that the tumor volume and weight of Ad5?Herg1?shRNA group were significantly smaller than of Ad5?control?shRNA group after 14 days and 5 weeks of inoculation, respectively (P<0.001).Moreover, knockdown of Herg1 inhibited the metastasis of OS cells.In mechanism, Herg1 protein interacted with NF2 protein. Knockdown of Herg1 significantly suppressed the expression levels of LATS1 and YAP protein, and promoted the phosphorylation of LATS1 and YAP in OS cells ( all P<0.001). Conclusion Our findings suggest that Herg1 participates in the proliferation and motility of OS cells and may serve as a potential therapeutic target for osteosarcoma patients.
		                        		
		                        		
		                        		
		                        	
10.2-deoxyglucose inhibits angiogenesis of rheumatoid arthritis via activating AMPK pathway.
Ying WANG ; Yingmei WEI ; Xiu CHENG ; Xiaojin SUN ; Linyan MA ; Yining SONG ; Jing ZHOU ; Fang WEI ; Hao LIU
Journal of Southern Medical University 2018;38(8):962-968
OBJECTIVETo observe the effects of 2-deoxyglucose inhibiting synovial pannus of adjuvant arthritis rats and to explore its potential mechanism of inhibiting angiogenesis by investigating proliferation, migration and matrigel tube formation assay .
METHODSThe effect of 2-DG on synovial pannus was evaluated by histopathology of HE staining; HUVEC proliferation was determined by CCK-8 method; migration of FLS were determined by transwell; matrigel tube formation assay was made for assessing tube number of HUVEC; p-AMPK and Bcl-2 were detected by Western blot assay; AMPK signaling pathway in HUVEC was inhibited by compound C, which is an inhibitor of AMPK activation.
RESULTS2-DG (200 mg/kg) obviously decreased appearance of synovial pannus ( < 0.01); , 2-DG (0.5 mmol/L and/or 5 mmol/L) obviously inhibited proliferation, migration and tube number of HUVEC ( < 0.01 or < 0.001), and its effects on HUVEC were reversed by using AMPK antagonist (Compound C); Western blot showed that 2-DG (5 mmol/L) increased expression of p-AMPK and decreased expression of Bcl-2 ( < 0.05).
CONCLUSIONSActivating AMPK pathway and decreasing expression of Bcl-2 may the potential mechanism by which 2-DG contributes to anti-angiogenesis and effects of inhibiting proliferation, migration and tube number of HUVEC.
            
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