1.Circular BANP knockdown inhibits the malignant progression of residual hepatocellular carcinoma after insufficient radiofrequency ablation
Guoming LI ; Jian KONG ; Shuying DONG ; Haigang NIU ; Shilun WU ; Wenbing SUN
Chinese Medical Journal 2022;135(13):1578-1587
		                        		
		                        			
		                        			Background::Circular RNAs (circRNAs) are endogenous non-coding RNAs, some of which have pathological roles. The current study aimed to explore the role of circRNA BTG3-associated nuclear protein (circ-BANP) binding with let-7f-5p and its regulation of the toll-like receptor 4 (TLR4)/signal transducer and activator of transcription 3 (STAT3) signaling pathway in residual hepatocellular carcinoma (HCC) after insufficient radiofrequency ablation (RFA).Methods::Circ-BANP, let-7f-5p, and TLR4 expressions in HCC samples were assessed using reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and Western blotting. Bioinformatics prediction, RNA pull-down assay, and dual luciferase reporter gene assay were used to analyze the relationships among circ-BANP, let-7f-5p, and TLR4. Huh7 cells were used to generate an in vitro model of residual HCC, defined as Huh7-H cells, which were transfected with either a plasmid or the sequence of circ-BANP, let-7f-5p, or TLR4. Expression of circ-BANP, let-7f-5p, and TLR4 mRNA was determined by RT-qPCR. TLR4, STAT3, p-STAT3, vascular endothelial growth factor A, vascular endothelial growth factor receptor-2, and epithelial-mesenchymal transformation (EMT)-related factors proteins were determined by Western blotting. Cell proliferation was determined by cell counting kit-8 and 5-Ethynyl-2’-deoxyuridine (EdU) assay and cell migration and invasion by Transwell assay. Animal studies were performed by inducing xenograft tumors in nude mice. Results::Circ-BANP and TLR4 mRNAs were upregulated in HCC tissues (the fold change for circ-BANP was 1.958 and that for TLR4 was 1.736 relative to para-tumors) and expression further increased following insufficient RFA (fold change for circ-BANP was 2.407 and that of TLR4 was 2.224 relative to para-tumors). Expression of let-7f-5p showed an opposite tendency (fold change for let-7f-5p in HCC tissues was 0.491 and that in tumors after insufficient RFA was 0.300 relative to para-tumors). Competitive binding of circ-BANP to let-7f-5p was demonstrated and TLR4 was identified as a target of let-7f-5p (P < 0.01). Knockdown of circ-BANP or elevation of let-7f-5p expression inhibited the TLR4/STAT3 signaling pathway, proliferation, invasion, migration, angiogenesis, and EMT in Huh7 and Huh7-H cells ( P < 0.01). The effects induced by circ-BANP knockdown were reversed by let-7f-5p inhibition. Overexpression of TLR4 reversed the impact of let-7f-5p upregulation on the cells ( P < 0.01). Silencing of circ-BANP inhibited the in vivo growth of residual HCC cells after insufficient RFA ( P < 0.01). Conclusions::Knockdown of circ-BANP upregulated let-7f-5p to inhibit proliferation, migration, and EMT formation in residual HCC remaining after insufficient RFA. Effects occur via regulation of the TLR4/STAT3 signaling pathway.
		                        		
		                        		
		                        		
		                        	
2.Endoscopic submucosal dissection for colorectal precancerous lesions and early cancer in the elderly over 80 years old
Enpan XU ; Bing LI ; Pinghong ZHOU ; Liqing YAO ; Qiang SHI ; Shilun CAI ; Zhipeng QI ; Di SUN ; Yunshi ZHONG
Chinese Journal of Digestive Endoscopy 2021;38(12):985-990
		                        		
		                        			
		                        			Objective:To evaluate the safety, efficacy and long-term survival of endoscopic submucosal dissection (ESD) for colorectal precancerous lesions and early cancer in the elderly over 80 years old.Methods:Clinical data of colorectal precancerous lesions and early cancer treated with ESD from January 2007 to December 2014 at Endoscopy Center of Zhongshan Hospital, Fudan University were retrospectively analyzed. A total of 721 patients with 778 lesions were included in this study. These patients were stratified by age: the super-elderly group (≥80 years old, 55 patients, 7.6%) and the non-super-elderly group (<80 years old, 666 patients, 92.4%). The outcomes of ESD, complication incidences, pathological characteristics, and long-term survival were compared between the two groups.Results:Except that the incidence of chronic concomitant diseases in the super-elderly group was significantly higher than that in the non-super-elderly group [54.5% (30/55) VS 31.5% (210/666), P<0.001], other baseline characteristics were not significantly different ( P>0.05). There were no significant differences in the complete resection rate [93.1%(54/58) VS 95.3%(686/720)], the R0 resection rate [89.7% (52/58) VS 93.2% (671/720)], the curative resection rate [84.5% (49/58) VS 90.3% (650/720)], the complication incidence [5.5% (3/55) VS 2.7%(18/666)], or the median hospitalization (2.98 days VS 2.54 days) between the two groups (all P>0.05). The three-year overall survival rates of the super-elderly group and non-super-elderly group were 95.8% and 98.0%, respectively, and the five-year overall survival rates were 85.1% and 97.4%, respectively. Conclusion:Colorectal ESD is safe and effective for elderly patients (age ≥80 years old) despite a significantly higher incidence of chronic concomitant diseases than that in the non-super-elderly patients.
		                        		
		                        		
		                        		
		                        	
3.Is drainage necessary in pelvic fracture patients with modified Stoppa approach?
Zhongzheng WANG ; Ao LI ; Ruipeng ZHANG ; Yingchao YIN ; Shilun LI ; Zheming GUO ; Zhiyong HOU ; Yingze ZHANG
Chinese Journal of Orthopaedics 2021;41(19):1412-1418
		                        		
		                        			
		                        			Objective:To investigate the effect of the placement of a drainage tube on the prognosis of patients with pelvic fractures treated by modified Stoppa approach.Methods:The medical records of patients with pelvic fractures treated with modified Stoppa approach from August 2012 to August 2017 were retrospectively analyzed. A total of 43 patients including 32 males and 11 females (mean age 47.6 years, range from16 to 69) were included in the study. According to Young-Burgess classification, there were 12 cases of Lateral Compression type LC-I type; 20 cases of Anterior and Posterior Compression type APC-I type and 11 cases of APC-II type. All patients were treated with modified Stoppa approach to reduce the fracture and fix with plate and screw. According to whether a drainage tube was placed during the operation, 22 cases were placed with a drainage tube (drainage group), and 21 cases were not placed with a drainage tube (non-drainage group). The main observation indicators were the intraoperative conditions, antibiotic application, incision suture removal time, postoperative body temperature change, hospital stay and clinical function (Harris score).Results:Wound infection was not observed in two groups. The duration of antibiotic use in the drainage group was 5.0 d (2.0, 8.0) d, and the non-drainage group was 4.0 d (2.0, 5.0) d, the difference was not statistically significant ( Z=1.161, P=0.924). The hospital stays of the two groups were 18.5 d (15.0, 24.3) d and 19.0 d (13.0, 26.0) d, respectively, and the difference was not statistically significant ( Z=0.542, P=0.591). The operation time was 150.2±52.4 min in the drainage group and 138.8±41.2 min in the non-drainage group, and the difference was not statistically significant ( t=0.791, P=0.433). The blood loss in the drainage group was 604.6±387.3 ml, and the non-drainage group was 581.0±275.0 ml. The difference was not statistically significant ( t=0.276, P=0.784). The postoperative body temperature changes of patients in the drainage group and non-drainage group were on day 1 (37.5±0.5 ℃ vs. 37.4±0.4 ℃, t=0.322, P>0.05), day 3 (37.1±0.4 ℃ vs. 37.0±0.4 ℃, t=0.286, P>0.05), day 5 (37.0± 0.3 ℃ vs. 36.8±0.2 ℃, t=2.127, P>0.05), on the 7th day (36.8±0.2 ℃ vs. 36.7±0.4 ℃, t=0.491, P>0.05), the difference was not statistically significant. The time for suture removal of surgical incision was 14.1±0.6 d in the drainage group and 13.9±0.6 d in the non-drainage group, and the difference was not statistically significant ( t=1.072, P=0.329). The Harris scores of the two groups were 96 (91, 100) points for the drainage group and 96 (93, 97) points for the non-drainage group, and the difference was not statistically significant ( Z=0.107, P=0.607). Conclusion:There is no significant influence of the application of drainage on recovery of wound or function for patients with pelvic surgery.
		                        		
		                        		
		                        		
		                        	
4.Experimental research on the trochlear dysplasia with bony and soft tissue model of patellar dislocation
Huijun KANG ; Yike DAI ; Shilun LI ; Jiangfeng LU ; Faquan LI ; Gang JI ; Yingzhen NIU ; Fei WANG
Chinese Journal of Orthopaedics 2020;40(6):372-380
		                        		
		                        			
		                        			Objective:To establish a model of patellar dislocation by femoral osteotomy or surgical release of medial retinaculum in immature rabbits, and observe morphological and trabecular microarchitectural changes in the trochlea.Methods:Forty rabbits at 3 months of age were included. All right knees underwent surgery, 20 knees were treated with femoral osteotomy and internal rotation of distal femur to increase femoral anteversion angle (Osteotomy group, OS group), and another 20 knees were treated with surgical release of medial retinaculum and overlap suture of lateral retinaculum (Soft tissue group, ST group). All left knees were acting as internal controls. Micro-CT scans for distal femur were acquired after 4 months post surgery. the height of Medial, central, and lateral trochlear, sulcus angle, and lateral and medial trochlear slope were measured to describe the trochlear morphology, and bone volume fraction (BV/TV), trabecular thickness (Tb.Th), trabecular number (Tb.N), trabecular spacing (Tb.Sp), and bone mineral density (BMD) were calculated to evaluate the microarchitectural structure. All parameters were compared between groups.Results:In OS group, one rabbit sustained a hip dislocation without patellar dislocation. Three knees developed complete patellar dislocation in daily flexion position, and the remaining 16 patellae were dislocated when the knee was placed in the maximal extension position. In ST group, 15 knees were complete patellar dislocation in daily flexion position, and 5 knees were without dislocation. A local boss was formed proximal to the entrance of the groove and the articular cartilage was smooth, and no obvious osteoarthritis was observed in OS group. In ST group no boss was formed, while obvious cartilage degeneration and defect was seen. Compared to the control group, the central trochlear height and sulcus angle were greater in both groups, but without significant difference between the two groups. The Tb.Th was increased in both medial and lateral condyle, and Tb.N was decreased in medial condyle compared with its control knees in OS group. The BV/TV, Tb.Th, Tb.N and BMD were decreased and Tb.Sp was increased in both medial and lateral condyle compared with its control knees in ST group. Compared to the OS group, the BV/TV, Tb.Th, Tb.N and BMD were smaller and Tb.Sp was greater in both medial and lateral condyle in ST group, with significant differences.Conclusion:The model of patellar dislocation could be successfully achieved by femoral rotational osteotomy to increase femoral anteversion or surgical release of medial retinaculum and overlap suture of lateral retinaculum, and subsequent morphological and trabecular microarchitectural changes in the trochlea are different. Different bony and soft tissue factors should be addressed for different patients with patellar dislocation in clinical practice.
		                        		
		                        		
		                        		
		                        	
5. The 474th case: anemia, ostealgia, proteinuria
Linyue WANG ; Xin LI ; Man SHEN ; Shilun CHEN ; Zhongxia HUANG
Chinese Journal of Internal Medicine 2020;59(2):161-164
		                        		
		                        			
		                        			 A 49-year-old woman was admitted to hospital with intermittent dizziness and fatigue for 7 years. The symptoms were aggravated and accompanied by bone pain for more than 4 months. She was referred to our hospital. Laboratory tests and imaging findings suggested that acquired Fanconi Syndrome (FS) was associated with smoldering multiple myeloma (MM). Renal biopsy and electron microscopy confirmed the diagnosis of proximal light chain tubular disease (LCPT). LCPT causes proximal tubular dysfunction, which is characterized by the cytoplasmic crystal deposition usually kappa monoclonal light chain in the proximal tubule. MM with FS and LCPT is less common in clinical practice because it is difficult to diagnose. This is a typical case focusing on the differential diagnosis of monoclonal gammopathy of renal significance(MGRS) such as LCPT and plasma cells diseases. 
		                        		
		                        		
		                        		
		                        	
6. Anatomic quadrilateral surface plate for the treatment of both-column acetabular fractures through Stoppa combined with iliac fossa approach
Ruipeng ZHANG ; Yingchao YIN ; Shilun LI ; Jialiang GUO ; Lin JIN ; Zhiyong HOU ; Yingze ZHANG
Chinese Journal of Orthopaedics 2019;39(13):781-788
		                        		
		                        			 Objective:
		                        			To explore the clinical outcome an anatomic quadrilateral surface plate for both-column acetabular fractures through the Stoppa combined with iliac fossa approach.
		                        		
		                        			Methods:
		                        			A retrospective study of the patients with both-column acetabular fractures treated with an anatomic quadrilateral surface plate through the Stoppa combined with iliac fossa approach from November 2015 to June 2017 was performed. Nineteen patients including 16 males and 3 females met the inclusion criteria. The mean age of included patients was 49.11 years (range, 25-68 years). There were 8 cases of fall injury, 7 cases of car accident injury and 4 cases of crushing injury. There were 11 cases of C1 type injury, 7 cases of C2 type, and 1 case of C3 type according to Tile classification. Patients included in this study were both-column acetabular fractures according to Letournel-Judet classification. There were six cases associated with posterior wall detachment. Minimally Stoppa combined with iliac fossa approach was performed to manage the displaced fragments. First, the anteromedially displaced iliopubic fragment was managed with a reconstruction plate through iliac fossa approach. Then, the medially displaced ischiadic fragment could be fixed with an anatomic quadrilateral surface plate through the Stoppa approach. The reduction quality was assessed by postoperative radiographs and CT scans according to the criteria proposed by Matta. The function of hip joint was assessed by the Merle d'Aubigné-Posteal score modified by Matta.
		                        		
		                        			Results:
		                        			The average operation time and blood loss were 115.26 min (range, 90-160 min) and 534.21 ml (range, 300-1000 ml), respectively. The mean length surgical incision was 18.82 cm (range, 16-20 cm). Average follow-up time was 19.84 months (range, 12-28 months). According to Matta criteria of reduction quality, anatomic reduction was obtained in 14 cases, and satisfactory reduction was gained in 3 cases, while unsatisfactory reduction was found in 2 cases (satisfactory rate=89.47%). All fractures healed well and mean healing time was 3.32 months (range, 3-5 months). The mean Merle d’Aubigné-Posteal score modified by Matta was 16.95 (range, 13-18), including 12 cases of excellent, 4 cases of good, and 3 cases of fair (satisfactory rate=84.21%). Intraoperative injury of lateral femoral cutaneous nerve developed in a case and obturator nerve damage occurred in another patient, respectively. Relevant symptoms were totally disappeared after two months’ conservative treatment (including neurotrophic therapy combined with adduction exercise of the affected thigh). Iliac fossa hematoma occurred in a case and relevant symptom was eliminated after puncture.
		                        		
		                        			Conclusion
		                        			Satisfactory clinical outcome of both-column acetabular fractures could be obtained by the anatomic quadrilateral surface plate through the Stoppa combined with iliac fossa approach. 
		                        		
		                        		
		                        		
		                        	
7. Application of dental floss traction-assisted endoscopic submucosa dissection to rectal neuroendocrine neoplasm
Qiang SHI ; Di SUN ; Yunshi ZHONG ; Meidong XU ; Bing LI ; Shilun CAI ; Zhipeng QI ; Zhong REN ; Hao ZHANG ; Yuanyuan YONG ; Liqing YAO ; Pinghong ZHOU
Chinese Journal of Gastrointestinal Surgery 2019;22(4):377-382
		                        		
		                        			 Objective:
		                        			To evaluate the safety and efficacy of dental floss traction-assisted endoscopic submucosal dissection (DFS-ESD) for rectal neuroendocrine neoplasm (NEN).
		                        		
		                        			Methods:
		                        			A retrospective cohort study was performed. Clinical data of rectal NEN patients undergoing ESD at Endoscopy Center of Zhongshan Hospital, Fudan University from January 2016 to December 2017 were retrospectively analyzed. Inclusion criteria: 1) age of 18 to 80 years old; 2) maximal diameter of lesions <1.5 cm; 3) tumor locating in the submucosa without invasion into the muscularis propria; 4) no enlarged lymph nodes around bowel and in abdominal cavity; 5) ESD requested actively by patients. A total of 37 patients were enrolled, including 23 male and 14 female cases with mean age of (56.0±11.3) years. All the lesions were single tumor of stage T1, and the mean size was 0.8±0.2(0.5-1.2) cm. Postoperative pathology revealed all samples as neuroendocrine tumors (NET). Seventeen patients received DFS-ESD treatment (DFS-ESD group) and 20 patient received conventional ESD treatment (conventional ESD group). In DFS-ESD group, after the mucosa was partly incised along the marker dots, the endoscopy was extracted, and the dental floss was tied to one arm of the metallic clip. When the endoscope was reinserted, the hemoclip was attached onto the incised mucosa; another hemoclip was attached onto normal mucosa opposite to the lesion in the same way. The submucosa was clearly exposed with the traction of dental floss and the resection could proceed. The conventional ESD group received the traditional ESD operation procedure. The operation time, modified operation time (remaining time after excluding the assembly time of dental floss traction in DFS-ESD group), en bloc resection rate, R0 resection rate, morbidity of operative complication, recurrence and metastasis were compared between two groups.
		                        		
		                        			Results:
		                        			The average tumor size was (0.8±0.2) cm in DFS-ESD group and (0.7±0.2) cm in conventional ESD group (
		                        		
		                        	
8.Application of artificial intelligence assisted diagnosis based on deep learning for early esophageal cancer
Shilun CAI ; Ayimukedisi YALIKONG ; Ran LI ; Bo YAN ; Liqing YAO ; Pinghong ZHOU ; Yunshi ZHONG
Chinese Journal of Digestive Endoscopy 2019;36(4):246-250
		                        		
		                        			
		                        			Objective To improve the detection rate of early esophageal cancer during endoscopy by construction of artificial intelligence assistant diagnosis system. Methods A total of 2400 esophageal images were collected from Zhongshan Hospital of Fudan University from January 2016 to December 2017, including 1200 images of early esophageal cancer and 1200 images of normal esophageal mucosa. The lesions in pictures were marked with rectangular box by using computer program. Among them, 2000 pictures were divided into the training set and 400 pictures into the test set. An assistant diagnostic model of early esophageal cancer was established by back propagation algorithm in computer deep learning. The training model was tested and the sensitivity and specificity of the system at different cut-off points in the test set was calculated. Receiver operating characteristic ( ROC) curve was used to evaluate the performance of the diagnostic model. Results The area under ROC curve ( AUC) of the auxiliary diagnostic model was 0. 9961. The sensitivity and specificity were satisfactory. Conclusion The deep learning model constructed in this study has good specificity, sensitivity and AUC value in the diagnosis of early esophageal cancer, and can assist endoscopists in real-time diagnosis in clinical examination.
		                        		
		                        		
		                        		
		                        	
9.Effects of endoscopic balloon dilatation on treatment of benign stenosis of colorectal anastomosis: a retrospective study
Bing LI ; Pinghong ZHOU ; Liqing YAO ; Meidong XU ; Zhong REN ; Qiang SHI ; Tao CHEN ; Shilun CAI ; Zhipeng QI ; Yunshi ZHONG
Chinese Journal of Digestive Endoscopy 2019;36(7):479-482
		                        		
		                        			
		                        			Objective To evaluate the efficacy and safety of endoscopic balloon dilation (EBD) on treatment of benign colorectal anastomotic stenosis.Methods Data of 36 patients with benign colorectal anastomotic stenosis undergoing EBD at Zhongshan Hospital from 2011 to 2015 were reviewed retrospectively.The number of dilation,complications rate,short-term effects and recurrence rate of stenosis were analyzed.Results Thirty-six patients had post-surgery anastomotic stenosis within 2-49 months (median 6 months),including 10 (27.8%) patients of grade 1 stenosis,15 (41.7%) of grade 2 stenosis,and 11 (30.5%) of grade 3 stenosis.The anastomotic distance from anus was 3-24 cm (median 6 cm).The 36 patients underwent 80 times of EBD with mean time of 2.22.Among them,69.4% (25/36) cases received 1-2 times and 30.6% (11/36) received 3 times or more.During the EBD operation,14 (17.5%) patients had minor bleeding,and intraoperative or postoperative perforation did not appeared.Postoperative obstructive symptoms were relieved in all patients.The anastomotic diameter was greater than 20 mm and EBD treatment was successful.Postoperative follow-up was 22-76 months (median 44 months).Four (11.1%) patients had recurrence of anastomotic benign stenosis at 7,11,18,and 63 months after the last time of EBD,respectively,and the symptoms were improved after the second treatment.Conclusion EBD is safe and effective in treating benign colorectal anastomotic stenosis,with better short-term and long-term outcomes.
		                        		
		                        		
		                        		
		                        	
10.Application of dental floss traction ? assisted endoscopic submucosa dissection to rectal neuroendocrine neoplasm
Qiang SHI ; Di SUN ; Yunshi ZHONG ; Meidong XU ; Bing LI ; Shilun CAI ; Zhipeng QI ; Zhong REN ; Hao ZHANG ; Yuanyuan YONG ; Liqing YAO ; Pinghong ZHOU
Chinese Journal of Gastrointestinal Surgery 2019;22(4):377-382
		                        		
		                        			
		                        			Objective To evaluate the safety and efficacy of dental floss traction?assisted endoscopic submucosal dissection (DFS?ESD) for rectal neuroendocrine neoplasm (NEN). Methods A retrospective cohort study was performed. Clinical data of rectal NEN patients undergoing ESD at Endoscopy Center of Zhongshan Hospital ,Fudan University from January 2016 to December 2017 were retrospectively analyzed. Inclusion criteria: 1) age of 18 to 80 years old; 2) maximal diameter of lesions <1.5 cm; 3) tumor locating in the submucosa without invasion into the muscularis propria; 4) no enlarged lymph nodes around bowel and in abdominal cavity; 5) ESD requested actively by patients. A total of 37 patients were enrolled,including 23 male and 14 female cases with mean age of (56.0±11.3) years. All the lesions were single tumor of stage T1,and the mean size was 0.8±0.2(0.5?1.2) cm. Postoperative pathology revealed all samples as neuroendocrine tumors (NET). Seventeen patients received DFS?ESD treatment (DFS?ESD group) and 20 patient received conventional ESD treatment (conventional ESD group). In DFS?ESD group,after the mucosa was partly incised along the marker dots ,the endoscopy was extracted,and the dental floss was tied to one arm of the metallic clip. When the endoscope was reinserted,the hemoclip was attached onto the incised mucosa; another hemoclip was attached onto normal mucosa opposite to the lesion in the same way. The submucosa was clearly exposed with the traction of dental floss and the resection could proceed. The conventional ESD group received the traditional ESD operation procedure. The operation time,modified operation time (remaining time after excluding the assembly time of dental floss traction in DFS?ESD group),en bloc resection rate,R0 resection rate, morbidity of operative complication,recurrence and metastasis were compared between two groups. Results The average tumor size was (0.8±0.2) cm in DFS?ESD group and (0.7±0.2) cm in conventional ESD group (t=0.425,P=0.673). According to postoperative pathological grading of rectal neuroendocrine neoplasm,13 were G1 and 4 were G2 in DFS?ESD group,while 17 cases were G1 and 3 cases were G2 in conventional ESD group without significant difference (P=0.680). There were no significant differences in baseline data between in the two groups (all P>0.05). All the basal resection margins were negative,the en bloc resection rate was 100% and the R0 resection rate was 100%. Pathological results showed tumor tissue close to the burning margin in 5 cases of conventional ESD group and in 2 cases of DFS?ESD group (P=0.416). The operation time was (17.9±6.6) minutes in conventional ESD group and (14.7±3.3) minutes in DFS?ESD group (t=1.776,P=0.084). The modified operation time of DFS?ESD group was (11.9±2.8) minutes,which was significantly shorter than (17.9±6.6) minutes in conventional ESD group (t=3.425, P=0.002). The hospital stay was (2.3 ± 0.6) days and (2.0 ± 0.5) days in conventional ESD group and DFS?ESD group,respectively,without significant difference (t=1.436,P=0.160). No patient was transferred to surgery,and no delayed bleeding or perforation occurred in either group. There was no recurrence or primary tumor?related death,and all the patients recovered well during a follow?up period of 14(1?24) months. Conclusion Dental floss traction?assisted ESD for rectal neuroendocrine neoplasm can simplify operation and ensure negative basal margin.
		                        		
		                        		
		                        		
		                        	
            
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