1.Mechanism of Buyang Huanwutang in Inhibiting Ferroptosis and Enhancing Neurological Function Recovery After Spinal Cord Injury via GPX4-ACSL4 Axis
Luchun XU ; Guozheng JIANG ; Yukun MA ; Jiawei SONG ; Yushan GAO ; Guanlong WANG ; Jiaojiao FAN ; Yongdong YANG ; Xing YU ; Xiangsheng TANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(5):20-30
ObjectiveTo explore the mechanism by which Buyang Huanwutang regulates the glutathione peroxidase 4 (GPX4)-acyl-CoA synthetase long-chain family member 4 (ACSL4) axis to inhibit ferroptosis and promote neurological functional recovery after spinal cord injury (SCI). MethodsNinety rats were randomly divided into five groups: sham operation group, model group, low-dose Buyang Huanwutang group (12.5 g·kg-1), high-dose Buyang Huanwutang group (25 g·kg-1), and Buyang Huanwutang + inhibitor group (25 g·kg-1 + 5 g·kg-1 RSL3). The SCI model was established by using the allen method. Tissue was collected on the 7th and 28th days after operation. Motor function was assessed by using the Basso-Beattie-Bresnahan (BBB) scale. Hematoxylin-eosin (HE), Nissl, and Luxol fast blue (LFB) staining were performed to observe spinal cord histopathology. Transmission electron microscopy was used to examine mitochondrial ultrastructure. Immunofluorescence staining was used to detect the number of NeuN-positive cells and the fluorescence intensity of myelin basic protein (MBP), GPX4, and ACSL4. Real-time fluorescent quantitative polymerase chain reaction (Real-time PCR) was used to analyze the mRNA expression of GPX4 and ACSL4. Enzyme linked immunosorbent assay (ELISA) was performed to measure the levels of reactive oxygen species (ROS), malondialdehyde (MDA), glutathione (GSH), and superoxide dismutase (SOD). Colorimetric assays were used to determine the iron content in spinal cord tissue. ResultsCompared to the sham operation group, the model group exhibited significantly reduced BBB scores (P<0.01), severe pathological damage in spinal cord tissue, and marked mitochondrial ultrastructural disruption. In addition, the model group showed a decrease in the number of NeuN-positive cells (P<0.01), reduced fluorescence intensity of MBP and GPX4 (P<0.01), lower levels of GSH and SOD (P<0.01), and downregulated mRNA expression of GPX4 (P<0.01). Moreover, compared to the sham operation group, the model group had elevated levels of ROS, MDA, and tissue iron content (P<0.01), along with increased fluorescence intensity and mRNA expression of ACSL4 (P<0.01). Compared with the model group and Buyang Huanwutang + inhibitor group, the Buyang Huanwutang group showed significantly improved BBB scores (P<0.05, P<0.01) and exhibited less severe spinal cord tissue damage, reduced edema and inflammatory cell infiltration, increased neuronal survival, and more intact myelin structures. Additionally, mitochondrial ultrastructure was significantly improved in the Buyang Huanwutang group. Compared to the model group and Buyang Huanwutang + inhibitor group, the Buyang Huanwutang group significantly increased the number of NeuN-positive cells and the fluorescence intensity of MBP (P<0.05, P<0.01). Furthermore, Buyang Huanwutang significantly increased the fluorescence intensity and mRNA expression of GPX4 (P<0.01) and decreased the fluorescence intensity and mRNA expression of ACSL4 (P<0.01) compared to the model group and Buyang Huanwutang + inhibitor group. Finally, the Buyang Huanwutang group significantly decreased ROS, MDA, and tissue iron content (P<0.01) and significantly increased GSH and SOD levels (P<0.01) compared to the model group and Buyang Huanwutang + inhibitor group. ConclusionBuyang Huanwutang inhibits ferroptosis through the GPX4/ACSL4 axis, reduces secondary neuronal and myelin injury and oxidative stress, and ultimately promotes the recovery of neurological function.
2.Bioinformatics analysis and identification of hub genes and their role in immune infiltration in osteoarthritis
Wei CAI ; Yukun ZHU ; Jianzhong XU
Chinese Journal of Tissue Engineering Research 2025;29(18):3747-3757
BACKGROUND:Low-grade,chronic inflammation is thought to play a central role in the pathogenesis of osteoarthritis.However,the specific molecular mechanisms are still unclear.OBJECTIVE:To screen and explore the potential hub genes and immune cell infiltration in osteoarthritis.METHODS:We merged data from the GSE206848 on the GPL570 and the GSE55235 and GSE55457 on the GPL96 to form the row dataset.Outlier samples were removed using weighted gene co-expression network analysis,followed by the identification of differentially expressed genes,and subsequent functional enrichment analysis of differentially expressed genes.Further,a protein-protein interaction network of differentially expressed genes was constructed,and hub genes were identified using two different algorithms in Cytoscape.Additionally,the CIBERSORT algorithm was employed to assess differences in immune cell infiltration proportions between osteoarthritis samples and normal controls.Finally,the diagnostic efficacy of hub genes for osteoarthritis was validated using quantitative reverse transcription polymerase chain reaction experiments conducted on synovial tissue samples collected from patients with osteoarthritis,in conjunction with the GSE12021 dataset from the GPL96 sequencing platform as an independent dataset.RESULTS AND CONCLUSION:After eliminating 5 outlier samples,we identified a total of 340 differentially expressed genes,comprising 159 up-regulated genes and 181 down-regulated genes.Six hub genes were obtained by weighted gene co-expression network analysis and Cytoscape.CIBERSORT analysis revealed a difference in the proportion of multiple types of immune cell infiltration in osteoarthritic tissues compared with normal tissues.Moreover,the expression levels of the six hub genes exhibited strong correlation with the relative proportion of multiple immune cells in osteoarthritis.The results of RT-qPCR indicated that the relative expression levels of the six genes were down-regulated relative to normal tissues.However,there was no significant difference in the expression of NFKBIA and PTGS2(P>0.05).Simultaneously,receiver operator characteristic curves in both the original and external datasets demonstrated that the six hub genes exhibited strong diagnostic capabilities for osteoarthritis(area under the curve>0.8).To conclude,four hub genes,CDKN1A,MYC,C-X-C motif chemokine ligand 2,and vascular endothelial growth factor A,are finally identified and may serve as molecular targets for future treatment by mediating immune response and inflammatory processes.
3.Prediction of Extrathyroidal Extension of Papillary Thyroid Carcinoma via Nomogram Model Based on Conventional Ultrasound and Contrast-Enhanced Ultrasound Features
Yiming CHENG ; Yu LIN ; Xu LI ; Taohua GOU ; Yan ZHANG ; Yukun LUO
Chinese Journal of Medical Imaging 2025;33(8):827-833,847
Purpose To construct a nomogram model based on conventional ultrasound(US)and contrast-enhanced ultrasound(CEUS)features for predicting extrathyroidal extension(ETE)of papillary thyroid carcinoma,and to evaluate its diagnostic performance.Materials and Methods A retrospective analysis of clinical and ultrasound data from 715 papillary thyroid carcinoma patients in the First Medical Center of Chinese PLA General Hospital from January 2017 to December 2022 was conducted.The patients were divided into two groups based on the presence or absence of ETE.Univariate and Multivariate analyses was performed to identify independent risk factors associated with ETE.Three models were established:clinical,clinical+US and clinical+US+CEUS.The nomogram of the best model was constructed and validated.Results The model based on clinical+US+CEUS features performed the best,the area under the curve was 0.885.Multivariate analysis indicated that older age(OR=1.029,95%CI 1.011-1.047),higher body mass index(OR=1.108,95%CI 1.049-1.171),capsular contact<25%(OR=4.716,95%CI 2.079-10.701),capsular contact 25%-50%(OR=21.320,95%CI 8.240-55.160),capsular contact>50%(OR=24.045,95%CI 6.792-85.126),nodules adjacent to the lateral side(OR=4.265,95%CI 1.366-13.318),nodules adjacent to the medial side(OR=6.416,95%CI 2.067-19.920)and interrupted capsular enhancement(OR=6.044,95%CI 3.588-10.180)were independent risk factors for ETE(all P<0.05).Decision curve analysis and clinical impact curve indicated high net benefit and strong clinical utility of the model.Ten-fold cross-validation showed good model stability.Conclusion The nomogram model constructed based on US and CEUS features demonstrates good predictive performance and holds significant clinical utility.
4."Textbook Outcome"and Influencing Factors in Patients with Pancreatic Ductal Adenocarcinoma Following Laparoscopic Pancreaticoduodenectomy:A Retrospective Cohort Study
Yakai YANG ; Shuai XU ; Chunhong ZHAO ; Yukun CAO ; Guangsheng YU ; Jun LIU
Cancer Research on Prevention and Treatment 2025;52(10):827-833
Objective To investigate the short-and long-term prognoses and the risk factors affecting"text-book outcome"(TO)after laparoscopic pancreaticoduodenectomy(LPD)for pancreatic ductal adenocar-cinoma(PDAC).Methods The clinical and follow-up data of patients diagnosed with PDAC and treated with LPD from January 2019 to Dec-ember 2022 were retrospectively anal-yzed.The prognosis was compared bet-ween TO and non-TO groups,and uni-variate and multivariate logistic regre-ssion analyses were used to identify independent prognostic factors for TO.Results A total of 284 patients were enrolled in this study,including 185 cases in the TO group and 99 cases in the non-TO group.The 1-,3-and 5-year overall survival(OS)rates of the TO and non-TO groups with PDAC were 87.3%vs.85.9%,52.5%vs.38.4%,and 18.0%vs.4.5%,respectively(P=0.020);the recurrence-free survival(RFS)rates were 74.1%vs.65.7%,27.1%vs.21.0%,and 10.0%vs.0%,respectively(P=0.042).Multivariate logistic regression analysis showed that operation time>360 min(OR=0.561,95%CI:0.321-0.979,P=0.042),intraoperative blood loss>400 ml(OR=0.392,95%CI:0.175-0.879,P=0.023),hard or tough texture of pancreas(OR=2.240,95%CI:1.247-4.022,P=0.007),and main pancreatic duct dia-meter>3 mm(OR=1.931,95%CI:1.126-3.312,P=0.017)were independent prognostic factors for TO.Conclusion After the learning curve,more than 60%of patients with PDAC can achieve TO after LPD.The chances of achieving TO are significantly reduced when the operation time>360 min,the intraoperative blood loss>400 ml,the texture of pancreas was soft,and the diameter of the main pancreatic duct>3 mm.
5.Analysis of the coding quality of lower extremity arteriosclerosis occlusion with thrombosis and endo-vascular intervention
Wenjia LI ; Deying KONG ; Yukun GOU ; Xinxin ZHANG ; Ying XU
Modern Hospital 2025;25(3):379-381
Objective By analyzing the causes of coding errors of lower extremity arteriosclerosis occlusion with throm-bosis and the endovascular intervention,explore the principle diagnosis and intervention coding rules to improve coding quality.Methods Inpatient medical records with the principle diagnosis codes of I70-I74 and I77 accompanied by endovascular interven-tional from January 1,2023 to December 31,2023 were retrieved from a tertiary hospital.A retrospective study was conducted to analyze the causes of coding errors.Results A total of 924 eligible cases were selected.There were 41 principle diagnosis cod-ing errors,with an error rate of 4.43%,among which the lower limb atherosclerosis coding had the highest error rate,accounting for 1.30%.There were 46 intervention coding errors,with an error rate of 4.98%,among which the percutaneous thrombectomy had the highest error rate.The main reason for coding errors was insufficient reading of medical record.Conclusion The coding error rate of the principle diagnosis of lower extremity arteriosclerosis occlusion and the endovascular intervention were relatively high.Therefore,it is necessary to continuously strengthen the training of clinicians and coders,including the coding rules and clinical knowledge of specialized disciplines,actively communicate between coders and clinicians,establish a quality control and assessment system,so as to improve the coding quality.
6.Antimicrobial resistance surveillance in the bacterial strains isolated from pediatric intensive care units in China:results from 2020 to 2022
Jing LIU ; Huiyuan YAN ; Gangfeng YAN ; Guoping LU ; Pan FU ; Chuanqing WANG ; Danqun JIN ; Wenjia TONG ; Chenyu ZHANG ; Jianli CHEN ; Yi LIN ; Jia LEI ; Yibing CHENG ; Qunqun ZHANG ; Kaijie GAO ; Yuanyuan CHEN ; Shufang XIAO ; Juan HE ; Li JIANG ; Huimin XU ; Yuxia LI ; Hanghai DING ; Hehe CHEN ; Yao ZHENG ; Qunying CHEN ; Ying WANG ; Hong REN ; Chenmei ZHANG ; Zhenjie CHEN ; Mingming ZHOU ; Yucai ZHANG ; Yiping ZHOU ; Zhenjiang BAI ; Saihu HUANG ; Lili HUANG ; Weiguo YANG ; Weike MA ; Qing MENG ; Pengwei ZHU ; Yong LI ; Yan XU ; Yi WANG ; Yanqiang DU ; Huijun CAI ; Bizhen ZHU ; Huixuan SHI ; Shaoxian HONG ; Yukun HUANG ; Meilian HUANG
Chinese Journal of Infection and Chemotherapy 2025;25(3):303-311
Objective This study aimed to investigate the antimicrobial resistance profiles of bacterial strains isolated from pediatric intensive care units(PICU)in China for better antimicrobial therapy.Methods Clinical isolates were collected from 17 institutions,including tertiary care children's hospitals and pediatric department of tertiary general hospitals in China from January 1,2020 to December 31,2022.Antimicrobial susceptibility testing was carried out according to a unified protocol using Kirby-Bauer method or automated systems.Results were interpreted according to the breakpoints released by the Clinical and Laboratory Standards Institute(CLSI)in 2020.Results A total of 10 688 isolates were collected,including gram-positive organisms(39.2%)and gram-negative organisms(60.8%).The top three organisms were S.aureus(13.6%,1 453/10 688),A.baumannii(10.0%,1 067/10 688),and coagulase-negative Staphylococcus(9.9%,1 058/10 688).Multi-drug resistant organisms(MDROs)were very common in children.The prevalence of methicillin-resistant Staphylococcus aureus(MRSA),carbapenem-resistant Enterobacterales(CRE),carbapenem-resistant E.coli,carbapenem-resistant K.pneumoniae(CRKP),carbapenem-resistant A.baumannii(CRAB),and carbapenem-resistant P.aeruginosa(CRPA)was 41.1%,19.4%,8.8%,30.9%,67.4%,and 28.8%,respectively.Overall,more than 50%of Enterobacteriales isolates were resistant to cephalosporins,while nearly 25%of Enterobacteriales isolates were resistant to carbapenems.MDROs were highly resistant to commonly used antibiotics.More than 80%of CRE and CRAB strains were resistant to all beta-lactam antibiotics.CRE and CRAB showed low resistance rates to tigecycline and polymyxin.CRPA showed lower resistance rates to piperacillin,beta-lactamase inhibitor combinations than the resistance rates to third and fourth generation cephalosporins.All of the Staphylococcus and Enterococcus isolates were susceptible to vancomycin and tigecycline.None of PRSP strains isolated from meningitis and nonmeningitis samples were resistant to rifampicin,vancomycin,or linezolid.The prevalence of β-lactamase-negative ampicillin-resistant(BLNAR)strains was 43.3%in Haemophilus influenzae.Conclusions MDROs were prevalent in PICU.It is necessary to establish an effective multidisciplinary team(MDT)to control the antimicrobial resistance.
7.Prediction of Extrathyroidal Extension of Papillary Thyroid Carcinoma via Nomogram Model Based on Conventional Ultrasound and Contrast-Enhanced Ultrasound Features
Yiming CHENG ; Yu LIN ; Xu LI ; Taohua GOU ; Yan ZHANG ; Yukun LUO
Chinese Journal of Medical Imaging 2025;33(8):827-833,847
Purpose To construct a nomogram model based on conventional ultrasound(US)and contrast-enhanced ultrasound(CEUS)features for predicting extrathyroidal extension(ETE)of papillary thyroid carcinoma,and to evaluate its diagnostic performance.Materials and Methods A retrospective analysis of clinical and ultrasound data from 715 papillary thyroid carcinoma patients in the First Medical Center of Chinese PLA General Hospital from January 2017 to December 2022 was conducted.The patients were divided into two groups based on the presence or absence of ETE.Univariate and Multivariate analyses was performed to identify independent risk factors associated with ETE.Three models were established:clinical,clinical+US and clinical+US+CEUS.The nomogram of the best model was constructed and validated.Results The model based on clinical+US+CEUS features performed the best,the area under the curve was 0.885.Multivariate analysis indicated that older age(OR=1.029,95%CI 1.011-1.047),higher body mass index(OR=1.108,95%CI 1.049-1.171),capsular contact<25%(OR=4.716,95%CI 2.079-10.701),capsular contact 25%-50%(OR=21.320,95%CI 8.240-55.160),capsular contact>50%(OR=24.045,95%CI 6.792-85.126),nodules adjacent to the lateral side(OR=4.265,95%CI 1.366-13.318),nodules adjacent to the medial side(OR=6.416,95%CI 2.067-19.920)and interrupted capsular enhancement(OR=6.044,95%CI 3.588-10.180)were independent risk factors for ETE(all P<0.05).Decision curve analysis and clinical impact curve indicated high net benefit and strong clinical utility of the model.Ten-fold cross-validation showed good model stability.Conclusion The nomogram model constructed based on US and CEUS features demonstrates good predictive performance and holds significant clinical utility.
8.Du Meridian electroacupuncture inhibits ferroptosis and promotes neurorepair in rats with acute cervical spinal cord injury
Rongyan SUN ; Luchun XU ; Guozheng JIANG ; Jiawei SONG ; Yukun MA ; Jiaojiao FAN ; Guanlong WANG ; Yongdong YANG ; Xing YU
Chinese Journal of Tissue Engineering Research 2025;29(29):6228-6236
BACKGROUND:Recent studies have shown that Du Meridian electroacupuncture has a unique effect on alleviating spinal cord injury,but the underlying mechanisms require further clarification.OBJECTIVE:To investigate the regulatory effects and the associated action mechanisms of Du Meridian electroacupuncture on ferroptosis after cervical spinal cord injury in rats.METHODS:One hundred SD rats were randomly divided into sham,model,Du Meridian electroacupuncture,RSL3,and Du Meridian electroacupuncture+RSL3 groups.The sham group underwent only laminectomy.The other four groups were subjected to cervical spinal cord injury by the Allen method.The Du Meridian electroacupuncture group received electroacupuncture after cervical spinal cord injury.The RSL3 group received intraperitoneal injections of glutathione peroxidase 4 inhibitor RSL3 after cervical spinal cord injury.The Du Meridian electroacupuncture+RSL3 group received both electroacupuncture and RSL3 intervention after cervical spinal cord injury.Samples were collected on postoperative days 7 and 28 to assess motor function,histological morphology,neuronal survival,glial scar formation,oxidative stress levels,Fe2+content,glutathione peroxidase 4,and long-chain acyl-CoA synthetase 4 expression.RESULTS AND CONCLUSION:(1)Finally,90 rats completed the follow-up experiment,with 18 rats in each group.(2)FLS and BBB scores were significantly higher in the Du Meridian electroacupuncture group compared with the model and Du Meridian electroacupuncture+RSL3 groups(P<0.05).(3)Compared with the model group,Du Meridian electroacupuncture improved cervical spinal cord tissue morphology and mitochondrial ultrastructure,while these effects were inhibited by RSL3.(4)Du Meridian electroacupuncture increased the expression of microtubule-associated protein 2,glutathione peroxidase 4,glutathione,and superoxide dismutase(P<0.05)and reduced the expression of glial fibrillary acidic protein,long-chain acyl-CoA synthetase 4,reactive oxygen species,malondialdehyde,and Fe2+compared with the model group(P<0.05).However,RSL3 reversed the inhibitory effects of Du Meridian electroacupuncture on ferroptosis,lipid peroxidation and oxidative stress.(5)The results suggest that Du Meridian electroacupuncture inhibits ferroptosis by regulating the glutathione peroxidase 4/long-chain acyl-CoA synthetase 4 axis,thereby reducing secondary neuronal damage and glial scar formation after cervical spinal cord injury and improving neurological function.
9.Relationship between traditional Chinese postpartum practices and postpartum depression
Shan CAO ; Jiajun XU ; Yukun KANG ; Peng WANG ; Min JIN
Sichuan Mental Health 2025;38(4):321-326
BackgroundPostpartum depression can affect the physical and mental health of mothers and the quality of parenting. Most Chinese women perform traditional postpartum practices (commonly known as "doing the month") after giving birth, while the existing findings are inconsistent and inconclusive regarding the potential of traditional Chinese postpartum practices to alleviate or exacerbate postpartum depression. ObjectiveTo explore the relationship between traditional Chinese postpartum practices and postpartum depression, so as to provide references for reducing the risk of postpartum depression. MethodsA total of 240 consecutive women who gave birth in the obstetrics department of the Mianyang Central Hospital and the Third Hospital of Mianyang from January to May 2024 were selected. Data were collected using Self-designed General Information Questionnaire, Chinese version of the Edinburgh Postnatal Depression Scale (EPDS), the Social Support Rating Scale (SSRS), the Patient Health Questionnaire-15 (PHQ-15), the Adherence to Doing-the-Month Practices questionnaire (ADP), and the Self-compiled Questionnaire on the Cognition of Doing-the-Month. The absolute value (A value) of the difference between scores of ADP and Cognition of Doing-the-Month Questionnaire was calculated to evaluate the degree of cognitive behavioral conflict of postpartum women. Pearson correlation analysis was performed to examine the correlations of EPDS score with SSRS score, PHQ-15 score, ADP total and dimensional scores, Cognition of Doing-the-Month Questionnaire total and dimensional scores, and A value. Logistic regression analysis was conducted to identify the protective and risk factors for developing postpartum depression. ResultsThe postpartum depression was detected in 22.50% of women. The postpartum women had a EPDS score of (6.21±5.00), ADP score of (70.05±20.57), SSRS score of (41.96±6.96), PHQ-15 score of (4.63±3.77), and Cognition of Doing-the-Month questionnaire score of (40.30±10.13). The A value was (0.65±0.58). Correlation analysis revealed that EPDS score was negatively correlated with the total ADP score and the four dimensional scores of the restrictions on social activities, diet, housework, and personal hygiene (r=-0.228, -0.146, -0.184, -0.275, -0.168, P<0.05 or 0.01), and positively correlated with the A value (r=0.161, P<0.05). Logistic regression analysis indicated that restriction on housework dimension in ADP was entered into the model (OR=0.930, 95% CI: 0.885~0.978). ConclusionThe restriction on housework dimension in traditional Chinese postpartum practices may be a protective factor against postpartum depression.
10.“Textbook Outcome” and Influencing Factors in Patients with Pancreatic Ductal Adenocarcinoma Following Laparoscopic Pancreaticoduodenectomy: A Retrospective Cohort Study
Yakai YANG ; Shuai XU ; Chunhong ZHAO ; Yukun CAO ; Guangsheng YU ; Jun LIU
Cancer Research on Prevention and Treatment 2025;52(10):827-833
Objective To investigate the short- and long-term prognoses and the risk factors affecting “textbook outcome” (TO) after laparoscopic pancreaticoduodenectomy (LPD) for pancreatic ductal adenocarcinoma (PDAC). Methods The clinical and follow-up data of patients diagnosed with PDAC and treated with LPD from January 2019 to December 2022 were retrospectively analyzed. The prognosis was compared between TO and non-TO groups, and univariate and multivariate logistic regression analyses were used to identify independent prognostic factors for TO. Results A total of 284 patients were enrolled in this study, including 185 cases in the TO group and 99 cases in the non-TO group. The 1-, 3- and 5-year overall survival (OS) rates of the TO and non-TO groups with PDAC were 87.3% vs. 85.9%, 52.5% vs. 38.4%, and 18.0% vs. 4.5%, respectively (P=0.020); the recurrence-free survival (RFS) rates were 74.1% vs. 65.7%, 27.1% vs. 21.0%, and 10.0% vs. 0%, respectively (P=0.042). Multivariate logistic regression analysis showed that operation time >360 min (OR=0.561, 95%CI: 0.321-0.979, P=0.042), intraoperative blood loss >400 ml (OR=0.392, 95%CI: 0.175-0.879, P=0.023), hard or tough texture of pancreas (OR=2.240, 95%CI: 1.247-4.022, P=0.007), and main pancreatic duct diameter >3 mm (OR=1.931, 95%CI: 1.126-3.312, P=0.017) were independent prognostic factors for TO. Conclusion After the learning curve, more than 60% of patients with PDAC can achieve TO after LPD. The chances of achieving TO are significantly reduced when the operation time >360 min, the intraoperative blood loss >400 ml, the texture of pancreas was soft, and the diameter of the main pancreatic duct >3 mm.

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