1.Uterine artery embolization for adenomyosis:evaluation of efficacy and analysis of predictive factors
Hao ZHANG ; Jinlian MA ; Xingwei SUN ; Mingyao WANG ; Yong JIN
Journal of Interventional Radiology 2025;34(7):766-771
Objective To evaluate the efficacy of uterine artery embolization(UAE)for the treatment of adenomyosis(AM)and to discuss the factors predicting efficacy.Methods The clinical data of 44 patients with AM,who received UAE at The Second Affiliated Hospital of Soochow University from January 2020 to August 2022,were retrospectively analyzed.Based on the MRI findings,serum CA125 levels and follow-up dysmenorrhea scores,the efficacy of UAE and the factors predicting efficacy were analyzed.Results A single UAE treatment was performed in all 44 patients with AM.Follow-up check with MRI at 12 months after treatment showed that the mean uterine volume decreased from preoperative(299.60±182.42)cm3 to postoperative(173.14±104.00)cm3,with a volume reduction of 36.64%(P<0.05).The dysmenorrhea score decreased from preoperative(7.70±2.60)points to postoperative(2.20+2.37)points,with a relief rate of 88.64%(P<0.05),and no severe adverse reactions occurred.Enhanced MRI demonstrated that in 30 patients(68.18%)the AM lesions presented as complete necrosis foci.In the patients whose preoperative lesions were completely necrotic,the mean preoperative serum CA125 level was(101.97±60.30)U/mL,the mean ADC value was(1.012+0.271)mm2/s,which were obviously lower than those in the patients whose preoperative lesions were incompletely necrotic(P<0.05).The mean T2 signal ratio(T2SR)value in the patients whose preoperative lesions were completely necrotic was(0.582±0.198),which was significantly higher than that in the patients whose preoperative lesions were incompletely necrotic(P<0.05).The analysis of receiver operating characteristic(ROC)curve showed that the preoperative area under the curve(AUC),sensitivity,and specificity for preoperative CA125 were 0.844,60%and 80%respectively;the AUC,sensitivity,and specificity for preoperative ADC were 0.760,80%and 65%respectively;and the AUC,sensitivity,and specificity for preoperative T2SR were 0.760,80%and 70%respectively.ADC<1.116 × 10-3 mm2/s,T2SR>0.479,and CA125<109.10 U/mL could be used as the reliable predictive factors for the efficacy of UAE treatment for AM.Conclusion UAE can effectively alleviate dysmenorrhea symptoms in patients with AM,and it is clinically safe and effective.ADC,T2SR,and CA125 can be used as the reliable predictive factors for the efficacy of UAE treatment for AM,which are helpful in predicting the lesion response and selecting the reasonable therapeutic options for patients.
2.Preparation and in vitro evaluation of an erythrocyte-based butyrylcholinesterase delivery system
Zhe WANG ; Changwen NING ; Huaying AN ; Xingwei JIANG ; Jun MA ; Fenghua GAO ; Pengyu LIU ; Yanan SUN ; Ru LI ; Jinlong LI ; Yuanyuan YUAN ; Qun YU
Military Medical Sciences 2025;49(6):458-464
Objective To develop an erythrocyte-based delivery system for butyrylcholinesterase(BChE)that is capable of prophylaxis against organophosphorus nerve agents.Methods Recombinant BChE was produced and analyzed for oligomerization via polyacrylamide gel electrophoresis(PAGE)and Western blotting.A modified hypotonic preswelling method was employed to prepare BChE-loaded erythrocytes.The drug loading capacity and encapsulation efficiency were quantified using enzyme-linked immunosorbent assay(ELISA).Catalytic activity was assessed in vitro with an activity detection kit.The system was characterized via scanning electron microscopy(SEM),flow cytometry and a hematology analyzer.Results Recombinant BChE predominantly existed as dimers(85%dimer,15%monomer).The optimized volume ratio of erythrocytes to hypotonic solution was determined as 1:7.Compared with native and empty erythrocytes,BChE-loaded erythrocytes exhibited significantly higher catalytic activity(P<0.001).The mean corpuscular volume of BChE-loaded erythrocytes increased(P<0.001),while the mean content of corpuscular hemoglobin and hemoglobin in erythrocytes per 100 mL decreased(P<0.001).SEM revealed no morphological differences(biconcave disc shape).Hypotonic preswelling moderately increased erythrocyte apoptosis(P<0.001),but no statistical difference was observed between BChE-loaded and hypotonic-treated erythrocytes(P>0.05).CD47 expression remained unchanged compared to native erythrocytes(P>0.05).Conclusion The modified hypotonic preswelling method can generate BChE-loaded erythrocytes that retain the characteristics of native erythrocytes while conferring catalytic activity,offering a novel strategy for clinical intervention against organophosphorus poisoning.
3.Construction of a nomogram model for personalized prediction of anal fistula occurrence after incision and drainage of perianal abscess
Changlin YAN ; Xingwei SUN ; Lu ZHAO
Journal of Clinical Surgery 2024;32(5):517-520
Objective To explore the risk factors of anal fistula after incision and drainage surgery for perianal abscess,and establish an individualized predictive nomogram model.Methods A retrospective analysis was conducted on the clinical data of 224 patients with perianal abscess who underwent incision and drainage surgery in Affiliated Hospital of Shanxi University of Chinese Medicine from May 2020 to January 2023,according to whether anal fistula occurred within 3 months after surgery,there were 169 cases in the non anal fistula group and 55 cases in the anal fistula group.Single factor method and multivariate Logistic regression analysis were applied to analyze the influencing factors of anal fistula after incision and drainage of perianal abscess,a nomogram risk model was constructed using independent risk factors to predict the occurrence of anal fistula after incision and drainage of perianal abscess,and the consistency and differentiation of the model were verified.Results The proportions of male,diabetes,deep abscesses,intestinal origin of pathogenic bacteria,and abscesses in anal fistula group were higher than those in non anal fistula group(P<0.05).Male,diabetes,deep abscess,intestinal origin of pathogenic bacteria,and history of abscess were independent risk factors for anal fistula after incision and drainage of perianal abscess(P<0.05).The ideal curve of the nomogram model fitted well with the correction curve,indicated that the measured values were basically consistent with the predicted values.The area under the receiver operating characteristic(ROC)curve was 0.946(95%CI=0.914-0.979),indicated that the column plot model has good predictive discrimination.Conclusion The independent risk factors for anal fistula after incision and drainage surgery of perianal abscess include male,diabetes,deep abscess,intestinal origin of pathogenic bacteria,and history of abscess.The construction of related nomogram model can guide clinical screening of high-risk groups to a certain extent.
4.Inguinal intranodal lymphangiography for abdominal and pelvic cavity traumatic lymphatic leakages
Mingqing ZHANG ; Xingwei SUN ; Jian ZHANG ; Xuming BAI ; Yong JIN
Chinese Journal of Interventional Imaging and Therapy 2024;21(5):281-284
Objective To observe the therapeutic effect and safety of inguinal intranodal lymphangiography in patients with abdominal and pelvic cavity traumatic lymphatic leakages.Methods Data of 12 patients with abdominal and pelvic cavity traumatic lymphatic leakages after ineffective conservative treatment and underwent inguinal intranodal lymphangiography were retrospectively analyzed.The clinical manifestations,therapeutic effects and complications were recorded.Results Totally 21 times of inguinal intranodal lymphangiography were performed in 12 patients,including 5 cases received 1 time,6 cases received 2 times and 1 case received 4 times,and the technical success rate of inguinal intranodal lymphangiography was 100%.After inguinal intranodal lymphangiography,7 cases(7/12,58.33%)were cured while 5 cases(5/12,41.67%)were not cured.The incidence of complications of inguinal intranodal lymphangiography was 14.29%(3/21),including chronic diarrhea after 2 times and puncture point pain in 1 case.No serious complication occurred.Conclusion Inguinal intranodal lymphangiography had certain therapeutic effect and high safety for abdominal and pelvic cavity traumatic lymphatic leakages.
5.Association Between the Aggregate Index of Systemic Inflammation and Albuminuria:A Cross-Sectional Study of National Health and Nutrition Examination Survey 2007-2018
Lirong SUN ; Xingwei HUO ; Shanshan JIA ; Xiaoping CHEN
Journal of Sichuan University (Medical Sciences) 2024;55(3):671-679
Objective Prior studies have established a connection between albuminuria and various inflammatory reactions,highlighting that an increase in C-reactive protein by 1 mg/L increases the likelihood of albuminuria by 2%.Recent investigations indicate a positive correlation between the systemic immune-inflammation index(SII)and increased urinary protein excretion.In addition,elevated levels of the systemic inflammatory response index(SIRI)also correlate with a higher prevalence of albuminuria.The aggregate index of systemic inflammation(AISI)offers a more comprehensive indicator of inflammation,providing an extensive assessment of systemic inflammatory status compared to SII and SIRI.Yet,the specific relationship between AISI and albuminuria remains unclear.This study aims to explore this association in U.S.adults.Methods We analyzed data from the National Health and Nutrition Examination Survey(NHANES)for 2007-2018,excluding pregnant women and individuals under 18.Cases with missing data on AISI,urinary albumin concentration,and other covariates were also excluded.AISI was computed using the formula:AISI=(platelet count×neutrophil count×monocyte count)/lymphocyte count.Albuminuria was defined as the urinary albumin-to-creatinine ratio exceeding 30 mg/g.Continuous variables were presented in the form of the mean±standard error,and categorical variables in percentages.We utilized weighted t-tests and chi-square tests for baseline comparisons.We applied weighted multivariable logistic regression and generalized additive models(GAM)to explore the association between AISI and albuminuria and to assess potential nonlinear relationships.Results The study included 32 273 participants,with an average age of(46.75±0.24)years old.The cohort comprised 48.73% males and 51.27% females.The prevalence of albuminuria was 9.64%.The average logarithmic value of log2AISI was 7.95±0.01,and were categorized into tertiles as follows:Quartile 1(Q1)(4.94 to 7.49),Q2(7.49 to 8.29),and Q3(8.29 to 10.85).As log2AISI increased,so did the prevalence of hypertension,diabetes,congestive heart failure,and albuminuria,all showing statistically significant increases(P<0.001).Similarly,the use of antihypertensive,lipid-lowering,and hypoglycemic drugs was also more prevalent(P<0.001).Statistically significant differences were observed across the three groups concerning age,race and ethnicity,formal education,alcohol consumption,smoking status,systolic and diastolic blood pressures,body mass index,estimated glomerular filtration rate,HbA1c,alanine aminotransferase,aspartate aminotransferase,albumin,creatinine,uric acid,and high-density lipoprotein cholesterol(P<0.05).However,no significant differences were noted in the total cholesterol or the sex ratios among the groups.The association between log2AISI and albuminuria was assessed using weighted multivariable logistic regression,and the detailed results are presented in Table 2.In model 1,without adjusting for covariates,each unit increase in log2AISI was associated with a 32% increase in the risk of albuminuria(odds ratio[OR]=1.32,95% confidence interval[CI]:1.27-1.38,P<0.001).Model 2 was adjusted for age,gender,race,and education level,and showed a similar trend,with each unit increase in log2AISI associated with a 31% increased risk(OR=1.31,95% CI:1.26-1.37,P<0.001).Model 3,which was further adjusted for all covariates,revealed that each unit increase in log2AISI was associated with a 20% increase in the risk of albuminuria(OR=1.20,95% CI:1.15-1.26,P<0.001).The study also transformed log2AISI from a continuous to a categorical variable for analysis.Compared with Q1,the risk of albuminuria in Q3,after adjusting for all covariates,significantly increased(OR=1.37,95% CI:1.22-1.55,P<0.001).Q2 also demonstrated a higher risk compared with Q1(OR=1.13,95% CI:1.06-1.36,P=0.004).The trend test indicated a dose-effect relationship between increasing log2AISI and the rising risk of albuminuria.GAM revealed a nonlinear relationship between log2AISI and albuminuria,with distinct trends noted between sexes.Segmented regression based on turning points showed significant effects among women,although the slope difference between the segments was not significant.In men,a significant threshold effect was observed;below the log2AISI of 7.25,increases in log2AISI did not enhance the risk of albuminuria,but above this threshold,the risk significantly increased.As part of a sensitivity analysis,weighted multivariable logistic regression was performed by changing the outcome variable to macroalbuminuria and adjusting for all covariates.The analysis showed that for every unit increase in log2AISI,the risk of developing macroalbuminuria increased by 31% (OR=1.31,95% CI:1.15-1.49,P<0.001).Compared with Q1,the risk of albuminuria in Q3 increased by 69% (OR=1.69,95% CI:1.27-2.25,P<0.001),and in Q2,it increased by 40% (OR=1.40,95% CI:1.03-1.92,P=0.030).Subgroup analysis and interaction results showed that the positive association between AISI and proteinuria risk was stronger in men than in women.Similarly,the association was stronger in people with hypertension compared with those with normal blood pressure,and higher in overweight people compared with those of normal weight.Furthermore,smokers and drinkers showed a stronger positive association between AISI and the risk of proteinuria than non-smokers and non-drinkers do.These results suggest that sex,blood pressure,body mass index,smoking,and alcohol consumption interact with AISI to influence the risk of proteinuria.Conclusion There is a robust positive association between AISI and increased risks of albuminuria in US adults.As log2AISI increases,so does the risk of albuminuria.However,further validation of this conclusion through large-scale prospective studies is warranted.
6.Application of computer-assisted navigation technology in the resection and reconstruction of mandibular ameloblastoma
Min LIU ; Enyi TANG ; Zhe LIU ; Sumeng GE ; Zhuhao WU ; Xingwei ZHANG ; Guowen SUN
STOMATOLOGY 2023;43(1):62-69
Objective:
Using computer-assisted navigation technology to guide the resection and reconstruction of mandibular ameloblastoma, evaluating its treatment effect.
Methods :
Twelve patients were selected from the Affiliated Stomatological Hospital of Nanjing University from January 2017 to May 2022. All 12 patients accepted same surgery which included resection of mandibular ameloblastoma and reconstruction by fibula musculocutaneous flap. Among them, 6 cases were included in the navigation group; 6 cases were in the non-navigation group. Advantages and disadvantages of computer-assisted navigation technology in this operation were evaluated with these cases.
Results:
The 12 operations were performed by the same operator. The average time for fixing the navigation bracket and performing navigation in the navigation group was about 15 minutes. Compared with the non-navigation group, the average operation time in the navigation group was shortened by about 10 minutes. In the navigation group, the mandible resection range matched the fibula musculocutaneous flap well, and the occlusal relationship recovered well.
Conclusion
Using the mandibular reference frame, under the guidance of computer-assisted navigation technology, the resection and reconstruction of mandibular ameloblastoma can be performed quickly and accurately.
7. Baicalein promotes laryngeal cancer cell death and inhibits invasion via miR-125b-5p/IRF4 axis
Jian WANG ; Yongdong SUN ; Xingwei ZHOU ; Lei LIU ; Long CHEN ; Xingke TONG ; Jiali ZHU
Chinese Journal of Clinical Pharmacology and Therapeutics 2023;28(11):1209-1218
AIM: To investigate the mechanism of baicalin-induced apoptosis in human laryngeal cancer cells. METHODS: AMC-HN-8 cells were selected for the study, and baicalin was applied to the cells at different concentrations (0, 10, 30, 100, and 300 μmol/L), and the half-inhibitory concentration (IC50) was measured by the CCK-8 method. Bax, cleaved-caspase-3, Cyto-c, IRF4 protein expression by protein blotting (Western blot); miR-125b-5p and IRF4 expression by RT-qPCR. Dual-luciferase reporter gene validation of Targetscan prediction (binding of miR-125b-5p to IRF4-3'UTR); apoptosis and necrosis inhibitors explore the way baicalein induces death in laryngeal cancer cells. AMC-HN-8 was then divided into blank group, baicalein (IC50), miR-125b-5p inhibitor group, baicalein + inhibitor NC group, baicalein+miR-125b-5p inhibitor group, and cell invasion and clone formation assays to detect cell invasion and proliferation ability, respectively. Apoptosis was detected by flow cytometry. RESULTS: Baicalein inhibited the proliferation of AMC-HN-8 cells in a dose-dependent manner with an IC50 value of 47.31 μmol/L. Compared with the blank group, 47.31 μmol/L baicalin induced apoptosis and inhibited cell invasion, while upregulating the expression of miR-125b-5p and suppressing the mRNA and protein levels of IRF4. The luciferase results showed that the miR-125b-5p mimic was able to inhibit the activity of the IRF4-3'UTR promoter relative to the NC mimic (mimic) group. Baicalein induces laryngeal cancer cell death in an apoptotic manner. In addition, the combination of 47.31 μmol/L baicalin and miR-125b-5p inhibitor affected the behavior of AMC-HN-8 cells, showing that compared with the blank group, the baicalin group showed a decrease in the number of cell clones, weakened invasion ability, and increased apoptosis; the miR - 125b-5p inhibitor group showed an increase in the number of cell clones, enhanced invasion ability and decreased apoptosis. The baicalin+ inhibitor NC group was consistent with baicalin, with no significant effect of inhibitor NC on cell behavior. The cloning, invasion, and apoptosis of cells in the baicalin+miR-125b-5p inhibitor group were intermediate between the baicalin and miR-125b-5p inhibitor groups. CONCLUSION: Baicalin inhibits the proliferation of AMC-HN-8 cells, and the mechanism may be related to miR-125b-5p targeting to inhibit the expression of IRF4, inducing the pro-apoptotic proteins Bax, cleaved-caspase3, and Cyto-c, and inhibiting the apoptosis suppressor protein Bcl-2 thereby inducing apoptosis.
8.Anticarin-β shows a promising anti-osteosarcoma effect by specifically inhibiting CCT4 to impair proteostasis.
Gan WANG ; Min ZHANG ; Ping MENG ; Chengbo LONG ; Xiaodong LUO ; Xingwei YANG ; Yunfei WANG ; Zhiye ZHANG ; James MWANGI ; Peter Muiruri KAMAU ; Zhi DAI ; Zunfu KE ; Yi ZHANG ; Wenlin CHEN ; Xudong ZHAO ; Fei GE ; Qiumin LV ; Mingqiang RONG ; Dongsheng LI ; Yang JIN ; Xia SHENG ; Ren LAI
Acta Pharmaceutica Sinica B 2022;12(5):2268-2279
Unlike healthy, non-transformed cells, the proteostasis network of cancer cells is taxed to produce proteins involved in tumor development. Cancer cells have a higher dependency on molecular chaperones to maintain proteostasis. The chaperonin T-complex protein ring complex (TRiC) contains eight paralogous subunits (CCT1-8), and assists the folding of as many as 10% of cytosolic proteome. TRiC is essential for the progression of some cancers, but the roles of TRiC subunits in osteosarcoma remain to be explored. Here, we show that CCT4/TRiC is significantly correlated in human osteosarcoma, and plays a critical role in osteosarcoma cell survival. We identify a compound anticarin-β that can specifically bind to and inhibit CCT4. Anticarin-β shows higher selectivity in cancer cells than in normal cells. Mechanistically, anticarin-β potently impedes CCT4-mediated STAT3 maturation. Anticarin-β displays remarkable antitumor efficacy in orthotopic and patient-derived xenograft models of osteosarcoma. Collectively, our data uncover a key role of CCT4 in osteosarcoma, and propose a promising treatment strategy for osteosarcoma by disrupting CCT4 and proteostasis.
9.Effect of negative pressure lithotomy on the incidence of infection related complications after percutaneous nephrolithotripsy
Xingwei YU ; Hongao TAN ; Yeqing SUN ; Yunqiu GAO
Chinese Journal of Postgraduates of Medicine 2020;43(8):734-737
Objective:To observe the incidence of infection related complications after percutaneous nephrolithotripsy.Methods:One hundred and forty-two patients with renal calculi who were treated in the First Affiliated Hospital of Zhejiang University of Traditional Chinese Medicine from January 2017 to December 2018 were divided into control group (71 cases) and experimental group (71 cases) by random number method. Among them, the control group was treated with common sheath, the experimental group was treated with negative pressure lithotomy, and the patients were followed up for 1 year after the operation to count the recurrence. The patients in the two groups were compared in terms of perioperative indexes, intraoperative complication rate, postoperative complication rate, recurrence rate in 1 year′s follow-up and quality of life in 1 year′s follow-up.Results:The operation time in two groups had no significant difference ( P>0.05). The amount of bleeding in the experimental group was significantly higher than that in the control group [(12.15 ± 1.06) ml vs. (13.03 ± 1.17) ml], the length of hospitalization was significantly shorter than that in the control group [(5.13 ± 0.67) d vs. (6.02 ± 0.78) d], and the differences were statistically significant ( P<0.01). The incidence of intraoperative complications in two groups had no significant difference ( P>0.05). The incidence of postoperative complications in the experimental group was significantly lower than that in the control group [1.41%(1/71) vs. 11.27%(8/71)], the recurrence rate in the follow-up period of 1 year was significantly lower than that in the control group [1.14%(1/71) vs. 9.86%(7/71)], and the differences were statistically significant ( P<0.05). The scores of postoperative World Health Organization Quality of Life Questionaire BREF (WHOQOL-BREF) of the two groups had no significant difference ( P>0.05). At 1 year′s follow-up, the scores of WHOQOL-BREF in the experimental group were significantly higher than those in the control group ( P<0.01). Conclusions:With the help of vacuum lithotripsy in percutaneous nephrolithotripsy, but the incidence of postoperative complications can be significantly reduced, the length of stay can be shortened, the follow-up recurrence can be reduced, and the quality of life can be improved.
10. Transanal lateral lymph node dissection surgery for 5 cases of mid-low rectal cancer
Ziwei ZENG ; Xingwei ZHANG ; Junji CHEN ; Liang HUANG ; Shuangling LUO ; Liang KANG
Chinese Journal of Gastrointestinal Surgery 2019;22(8):781-785
Objective:
To evaluate the feasibility and safety of transanal lateral lymph node dissection for mid-low rectal cancer.
Methods:
A descriptive case series research method was used. Clinical and pathological data of 5 mid-low rectal cancer patients who underwent transanal lateral lymph node dissection at Department of Colorectal Surgery, the Sixth Affiliated Hospital of Sun Yat-sen University from November 2018 to May 2019 were retrospectively collected and analyzed. Of 5 cases, 4 were male and 1 was female with mean age of (43.2±13.2) years and mean body mass index of (21.2±2.6) kg/m2; the mean diameter of tumor was (3.2±2.4) cm; the mean distance between tumor and anus was (6.3±2.5) cm; 3 received preoperative neoadjuvant chemotherapy. In preoperative TNM staging, 2 cases were T3N1M0, 1 was T3cN2aM0, 1 was T3cN2bM0, and 1 was T2N1M0. All the patients had no intestinal obstruction before operation. Surgical methods: (1) total mesorectal excision: using general transanal and transabdominal methods to mobilize and resect total mesorectum, and dissect No.252, No.253 lymph nodes; (2) transanal lateral lymph node dissection: dissect the adipose lymphoid tissue on the surface of the iliococcygeal muscle, the coccygeal muscle, and the obturator muscle (the No.283 lymph nodes) upward, and dissect No.263d and No.263p lymph nodes with fat tissue sequentially till the bifurcation of the internal and external iliac artery; (3) take out the specimen from anus, and make anastomosis between proximal colon and anal canal. Intraoperative and postoperative variables was observed.
Results:
All the 5 patients completed surgery successfully, and no patient needed to convert to open approach. The mean operative time was (295.6±97.7) minutes, and the median intraoperative blood loss was 70 (50-500) ml. The mean length of specimen was (12.9±3.0) cm, and the mean number of harvested lymph node was 30.4±9.9. The positive lateral lymph nodes were founder in 4 patients. The median distance between tumor and distal resection margin was 1.5 (1.2-8.0) cm. The resection margin in all the patients was negative. The mean time to postoperative flatus was (4.2±1.6) days, the mean postoperative spontaneous urination was (3.0±1.9) days, time to drainage tube removal was (5.6±1.9) days, and the mean postoperative hospital stay was (9.4±2.1) days. The postoperative TNM staging by pathology was 1 case with T1N0M0, 1 with T2N1M0, 1 with T3N2bM0, and 2 with T3N2M0. Five patients were moderately differentiated adenocarcinoma. Only 1 patient developed postoperative abdominal bleeding, who was healed after conservative treatment. The other 4 patients did not develop any perioperative complications, such as incision infection, presacral abscess, pelvic abscess, anastomotic leakage, or anastomotic stricture. Four patients underwent postoperative chemotherapy. All the patients were followed up for 2 to 28 weeks after surgery and they all felt well. The patients with stoma had fluent bowel.
Conclusions
Transanal lateral lymph node dissection is feasible and safe in the treatment of mid-low rectal cancer, which can achieve the purpose of extended radical resection of mid-low movement rectal cancer. Moreover, this procedure is a new method to treat rectal cancer patients with lateral lymph node metastasis.

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