1.Multimodal ultrasound of the testis in differentiating subtypes of spermatogenic dysfunction:a preliminary study
Tingting LEI ; Gaoxiang FAN ; Penglin ZOU ; Chao JIA ; Hongmei LIANG ; Jun LIN ; Rong WU ; Lianfang DU ; Zheng LI ; Qiusheng SHI
Chinese Journal of Ultrasonography 2025;34(8):678-685
Objective:To analyze the imaging features of testicular conventional ultrasonography(US),shear wave elastography(SWE),and contrast-enhanced ultrasonography(CEUS)in subtypes of spermatogenic dysfunction(focal,arrest,and exhausted),and to evaluate the diagnostic efficacy of testicular multimodal ultrasound.Methods:A prospective study enrolled 310 spermatogenic dysfunction patients(focal type group: n=77,arrest type group: n=20,exhaustive type group: n=213)and 30 healthy volunteers(control group)at Shanghai General Hospital between October 2023 and December 2024. All patients underwent preoperative testicular US,SWE,and CEUS examinations,followed by microdissection testicular sperm extraction. Ultrasound parameters were compared among groups,and receiver operating characteristic(ROC)curves were plotted to assess the diagnostic efficacy of multimodal ultrasound for different subtypes. Results:①Testicular volume:The arrest group exhibited significantly greater volume compared to the focal group and the exhausted group(all P<0.05). No statistically significant difference was observed between the arrest group and the normal group( P>0.05). ②Mean Young's modulus(Emean):The arrest group exhibited significantly lower Emean compared with both the focal and exhausted groups(all P<0.05),but showed no significant difference versus the normal group( P>0.05). ③CEUS quantitative parameters:The arrival time(AT)and time to peak(TTP)in the exhausted group were higher than those in the normal group,while the peak intensity(PI)and area under the time-intensity curve(AUC)were lower(all P<0.05). No significant differences were found in AT,TTP,or AUC among the three spermatogenic dysfunction subgroups(all P>0.05). ④CEUS perfusion patterns:The focal group predominantly exhibited a mottled pattern(70.13%,54/77). Both the arrest group(90.00%,18/20)and the normal group(93.33%,28/30)predominantly exhibited an rich pattern. The exhausted group predominantly exhibited a sparse pattern(56.34%,120/213). ⑤ROC curve analysis showed that for differentiating the arrest group from the focal type group and the exhaustive type group,the combination of Emean,testicular volume,and CEUS perfusion pattern yielded an AUC of 0.931,with a sensitivity of 95.00% and specificity of 85.86%. For differentiating the focal type group from the exhaustive type group,the combination of testicular echogenicity,CEUS perfusion pattern,and Emean yielded an AUC of 0.833,with a sensitivity of 81.82% and specificity of 75.12%. Conclusions:Multimodal ultrasonography can comprehensively characterize the imaging features of different subtypes of spermatogenic dysfunction,holding promise as a noninvasive predictive tool.
2.Construction and evaluation of gastrointestinal bleeding nomogram after laparoscopic pancreaticoduodenectomy for patients with periampullary space occupying lesion
Shuai WANG ; Dongrui LI ; Jianhua LIU ; Chengxu DU ; Qiusheng LI ; Jianzhang QIN ; Haitao LYU
Chinese Journal of Hepatobiliary Surgery 2025;31(3):182-187
Objective:To construct a nomogram model for predicting the risk factors of gastrointestinal bleeding following laparoscopic pancreaticoduodenectomy (LPD) based on relevant risk factors and evaluate its predictive value.Methods:A retrospective analysis was conducted on 466 patients with periampullary space occupying lesion who underwent LPD at the Department of Hepatobiliary Surgery, the Second Hospital of Hebei Medical University, from January 2021 to December 2024. Among them, there were 284 males and 182 females, aged (59.9±10.7) years. Patients were randomly divided into a training cohort ( n=326) and a validation cohort ( n=140) using a random number table (7: 3 ratio). Based on whether patients suffered gastrointestinal bleeding, the training cohort was further stratified into a gastrointestinal bleeding group ( n=23) and control group ( n=303). Univariate and multivariate logistic regression analyses were performed to identify independent risk factors for gastrointestinal bleeding. A nomogram was constructed based on multivariate results, and its predictive performance was evaluated by receiver operating characteristic (ROC) curves, calibration curves, and decision curve analysis (DCA). Results:Compared to the control group, the gastrointestinal bleeding group exhibited significantly higher age, higher rates of postoperative pancreatic fistula (POPF) and intra-abdominal infection, along with lower body mass index, and lower levels of fibrinogen and albumin (all P<0.05). Multivariate analysis identified age ( OR=1.065, 95% CI: 1.002-1.132), fibrinogen ( OR=0.486, 95% CI: 0.243-0.969), albumin ( OR=0.840, 95% CI: 0.741-0.953), POPF ( OR=4.299, 95% CI: 1.348-13.716), and postoperative intra-abdominal infection ( OR=6.352, 95% CI: 1.476-27.341) as independent predictors of gastrointestinal bleeding (all P<0.05). The nomogram demonstrated robust discrimination, with an AUC of 0.861 (95% CI: 0.784-0.939), sensitivity of 82.6%, and specificity of 82.2% in the training cohort. In the validation cohort, the AUC was 0.824 (95% CI: 0.675-0.973), with sensitivity and specificity of 80.0% and 83.8%, respectively. Calibration curves indicated excellent agreement between predicted and observed outcomes. DCA revealed superior net clinical benefit of the nomogram over " treat-all" or " treat-none" strategies within threshold probabilities of 0-0.9 (training) and 0-0.75 (validation). Conclusion:The nomogram based on age, fibrinogen, albumin, POPF, and intra-abdominal infection provides accurate prediction of gastrointestinal bleeding after LPD and demonstrates high clinical utility for risk stratification and decision-making in periampullary space occupying lesion patients.
3.Telocytes-Mediated Effects and Mechanisms of Anointing and Massage Therapy Using Oligopeptide-Herbal Medicine Composite Against Obesity in Rats
Yifan LUO ; Zhenwei ZHANG ; Lu MEI ; Yeping SHI ; Yitong XING ; Zeqi ZHANG ; Chuxin LI ; Chunxia HAN ; Pingshun YANG ; Qiusheng CHEN
Laboratory Animal and Comparative Medicine 2025;45(5):551-560
Objective To investigate the weight loss efficacy and mechanisms of oligopeptide-herbal medicine composite,and to provide new approaches for obesity treatment.Methods Twenty-three SPF female SD rats were randomly divided into control group(n=3)and modeling group(n=20).The control group was fed an ordinary diet for 6 consecutive weeks,and the modeling group was fed a high-fat diet for 6 consecutive weeks to establish a simple obesity rat model.After successful modeling,the modeling group was randomly divided into model group(n=10)and treatment group(n=10).The treatment group started anoint-and-massage therapy with oligopeptide-herbal medicine composite(3 g per rat per time,once a day,20 min each time),which was recorded as day 1.The control group and model group were not treated with oligopeptide-herbal medicine composite but received the same massage,and continued to be fed ordinary feed and high-fat feed respectively.Ten rats in the model group and ten in the treatment group were each divided into 3 groups,with 3,3,and 4 rats in each group.On days 3,6,and 9 after treatment,the rats in the three groups were weighed and cardiac blood collection was performed after isoflurane respiratory anesthesia.After euthanasia by cardiac bloodletting,abdominal subcutaneous adipose tissue(aSAT)and perirenal white adipose tissue(pWAT)samples were collected and weighed.Serum triglycerides(TG)and high-density lipoprotein cholesterol(HDL-C)levels were measured.Image J software was used to measure aSAT thickness and the diameter and area of perirenal white adipocytes and abdominal subcutaneous adipocytes.Immunofluorescence technique was used to observe the number of telocytes(TCs),cell junctions and exosomes per unit area of aSAT.Transmission electron microscopy was used to measure the length of telopodes(Tps)of dermal and subcutaneous TCs in abdominal skin,and to observe the distribution of exosome vesicles,rough endoplasmic reticulum,mitochondria,and cell junctions.Results Compared with the control group,the body weight of rats in the modeling group increased significantly(P<0.05).Compared with the control group,rats in the model group showed a significant increase in serum TG level,a significant decrease in HDL-C level,and a significant increase in pWAT mass(all P<0.05).Compared with the model group,the treatment group showed a significantly increased rate of body weight reduction,significantly decreased pWAT mass,significantly decreased serum TG level,and significantly increased HDL-C level(all P<0.05).Compared with the model group,the treatment group showed significant reductions in aSAT thickness,as well as cell diameter and area in both aSAT and pWAT(all P<0.05).Compared with the model group,the treatment group showed significantly increased number of TCs per unit area of aSAT,number of exosomes from TCs in aSAT,and Tps length(all P<0.05).The treatment group showed an increasing trend in the numbers of mitochondria,rough endoplasmic reticulum,and cell junctions in TCs.Conclusion Oligopeptide-herbal medicine composite applied via anoint-and-massage therapy effectively improves obesity-related symptoms such as dyslipidemia and fat accumulation in obese rats by regulating TCs and their intercellular communication.
4.Telocytes-Mediated Effects and Mechanisms of Anointing and Massage Therapy Using Oligopeptide-Herbal Medicine Composite Against Obesity in Rats
Yifan LUO ; Zhenwei ZHANG ; Lu MEI ; Yeping SHI ; Yitong XING ; Zeqi ZHANG ; Chuxin LI ; Chunxia HAN ; Pingshun YANG ; Qiusheng CHEN
Laboratory Animal and Comparative Medicine 2025;45(5):551-560
Objective To investigate the weight loss efficacy and mechanisms of oligopeptide-herbal medicine composite,and to provide new approaches for obesity treatment.Methods Twenty-three SPF female SD rats were randomly divided into control group(n=3)and modeling group(n=20).The control group was fed an ordinary diet for 6 consecutive weeks,and the modeling group was fed a high-fat diet for 6 consecutive weeks to establish a simple obesity rat model.After successful modeling,the modeling group was randomly divided into model group(n=10)and treatment group(n=10).The treatment group started anoint-and-massage therapy with oligopeptide-herbal medicine composite(3 g per rat per time,once a day,20 min each time),which was recorded as day 1.The control group and model group were not treated with oligopeptide-herbal medicine composite but received the same massage,and continued to be fed ordinary feed and high-fat feed respectively.Ten rats in the model group and ten in the treatment group were each divided into 3 groups,with 3,3,and 4 rats in each group.On days 3,6,and 9 after treatment,the rats in the three groups were weighed and cardiac blood collection was performed after isoflurane respiratory anesthesia.After euthanasia by cardiac bloodletting,abdominal subcutaneous adipose tissue(aSAT)and perirenal white adipose tissue(pWAT)samples were collected and weighed.Serum triglycerides(TG)and high-density lipoprotein cholesterol(HDL-C)levels were measured.Image J software was used to measure aSAT thickness and the diameter and area of perirenal white adipocytes and abdominal subcutaneous adipocytes.Immunofluorescence technique was used to observe the number of telocytes(TCs),cell junctions and exosomes per unit area of aSAT.Transmission electron microscopy was used to measure the length of telopodes(Tps)of dermal and subcutaneous TCs in abdominal skin,and to observe the distribution of exosome vesicles,rough endoplasmic reticulum,mitochondria,and cell junctions.Results Compared with the control group,the body weight of rats in the modeling group increased significantly(P<0.05).Compared with the control group,rats in the model group showed a significant increase in serum TG level,a significant decrease in HDL-C level,and a significant increase in pWAT mass(all P<0.05).Compared with the model group,the treatment group showed a significantly increased rate of body weight reduction,significantly decreased pWAT mass,significantly decreased serum TG level,and significantly increased HDL-C level(all P<0.05).Compared with the model group,the treatment group showed significant reductions in aSAT thickness,as well as cell diameter and area in both aSAT and pWAT(all P<0.05).Compared with the model group,the treatment group showed significantly increased number of TCs per unit area of aSAT,number of exosomes from TCs in aSAT,and Tps length(all P<0.05).The treatment group showed an increasing trend in the numbers of mitochondria,rough endoplasmic reticulum,and cell junctions in TCs.Conclusion Oligopeptide-herbal medicine composite applied via anoint-and-massage therapy effectively improves obesity-related symptoms such as dyslipidemia and fat accumulation in obese rats by regulating TCs and their intercellular communication.
5.Multimodal ultrasound of the testis in differentiating subtypes of spermatogenic dysfunction:a preliminary study
Tingting LEI ; Gaoxiang FAN ; Penglin ZOU ; Chao JIA ; Hongmei LIANG ; Jun LIN ; Rong WU ; Lianfang DU ; Zheng LI ; Qiusheng SHI
Chinese Journal of Ultrasonography 2025;34(8):678-685
Objective:To analyze the imaging features of testicular conventional ultrasonography(US),shear wave elastography(SWE),and contrast-enhanced ultrasonography(CEUS)in subtypes of spermatogenic dysfunction(focal,arrest,and exhausted),and to evaluate the diagnostic efficacy of testicular multimodal ultrasound.Methods:A prospective study enrolled 310 spermatogenic dysfunction patients(focal type group: n=77,arrest type group: n=20,exhaustive type group: n=213)and 30 healthy volunteers(control group)at Shanghai General Hospital between October 2023 and December 2024. All patients underwent preoperative testicular US,SWE,and CEUS examinations,followed by microdissection testicular sperm extraction. Ultrasound parameters were compared among groups,and receiver operating characteristic(ROC)curves were plotted to assess the diagnostic efficacy of multimodal ultrasound for different subtypes. Results:①Testicular volume:The arrest group exhibited significantly greater volume compared to the focal group and the exhausted group(all P<0.05). No statistically significant difference was observed between the arrest group and the normal group( P>0.05). ②Mean Young's modulus(Emean):The arrest group exhibited significantly lower Emean compared with both the focal and exhausted groups(all P<0.05),but showed no significant difference versus the normal group( P>0.05). ③CEUS quantitative parameters:The arrival time(AT)and time to peak(TTP)in the exhausted group were higher than those in the normal group,while the peak intensity(PI)and area under the time-intensity curve(AUC)were lower(all P<0.05). No significant differences were found in AT,TTP,or AUC among the three spermatogenic dysfunction subgroups(all P>0.05). ④CEUS perfusion patterns:The focal group predominantly exhibited a mottled pattern(70.13%,54/77). Both the arrest group(90.00%,18/20)and the normal group(93.33%,28/30)predominantly exhibited an rich pattern. The exhausted group predominantly exhibited a sparse pattern(56.34%,120/213). ⑤ROC curve analysis showed that for differentiating the arrest group from the focal type group and the exhaustive type group,the combination of Emean,testicular volume,and CEUS perfusion pattern yielded an AUC of 0.931,with a sensitivity of 95.00% and specificity of 85.86%. For differentiating the focal type group from the exhaustive type group,the combination of testicular echogenicity,CEUS perfusion pattern,and Emean yielded an AUC of 0.833,with a sensitivity of 81.82% and specificity of 75.12%. Conclusions:Multimodal ultrasonography can comprehensively characterize the imaging features of different subtypes of spermatogenic dysfunction,holding promise as a noninvasive predictive tool.
6.Construction and evaluation of gastrointestinal bleeding nomogram after laparoscopic pancreaticoduodenectomy for patients with periampullary space occupying lesion
Shuai WANG ; Dongrui LI ; Jianhua LIU ; Chengxu DU ; Qiusheng LI ; Jianzhang QIN ; Haitao LYU
Chinese Journal of Hepatobiliary Surgery 2025;31(3):182-187
Objective:To construct a nomogram model for predicting the risk factors of gastrointestinal bleeding following laparoscopic pancreaticoduodenectomy (LPD) based on relevant risk factors and evaluate its predictive value.Methods:A retrospective analysis was conducted on 466 patients with periampullary space occupying lesion who underwent LPD at the Department of Hepatobiliary Surgery, the Second Hospital of Hebei Medical University, from January 2021 to December 2024. Among them, there were 284 males and 182 females, aged (59.9±10.7) years. Patients were randomly divided into a training cohort ( n=326) and a validation cohort ( n=140) using a random number table (7: 3 ratio). Based on whether patients suffered gastrointestinal bleeding, the training cohort was further stratified into a gastrointestinal bleeding group ( n=23) and control group ( n=303). Univariate and multivariate logistic regression analyses were performed to identify independent risk factors for gastrointestinal bleeding. A nomogram was constructed based on multivariate results, and its predictive performance was evaluated by receiver operating characteristic (ROC) curves, calibration curves, and decision curve analysis (DCA). Results:Compared to the control group, the gastrointestinal bleeding group exhibited significantly higher age, higher rates of postoperative pancreatic fistula (POPF) and intra-abdominal infection, along with lower body mass index, and lower levels of fibrinogen and albumin (all P<0.05). Multivariate analysis identified age ( OR=1.065, 95% CI: 1.002-1.132), fibrinogen ( OR=0.486, 95% CI: 0.243-0.969), albumin ( OR=0.840, 95% CI: 0.741-0.953), POPF ( OR=4.299, 95% CI: 1.348-13.716), and postoperative intra-abdominal infection ( OR=6.352, 95% CI: 1.476-27.341) as independent predictors of gastrointestinal bleeding (all P<0.05). The nomogram demonstrated robust discrimination, with an AUC of 0.861 (95% CI: 0.784-0.939), sensitivity of 82.6%, and specificity of 82.2% in the training cohort. In the validation cohort, the AUC was 0.824 (95% CI: 0.675-0.973), with sensitivity and specificity of 80.0% and 83.8%, respectively. Calibration curves indicated excellent agreement between predicted and observed outcomes. DCA revealed superior net clinical benefit of the nomogram over " treat-all" or " treat-none" strategies within threshold probabilities of 0-0.9 (training) and 0-0.75 (validation). Conclusion:The nomogram based on age, fibrinogen, albumin, POPF, and intra-abdominal infection provides accurate prediction of gastrointestinal bleeding after LPD and demonstrates high clinical utility for risk stratification and decision-making in periampullary space occupying lesion patients.
7.Handling health emergency in public hospitals
Zhijun LUO ; Dayue LIU ; Jiang CHEN ; Liangcheng XIAO ; Li'an LI ; Yuyao WANG ; Qingliang ZENG ; Qiusheng LIU
Modern Hospital 2024;24(2):293-296
Public hospitals play a dominant role in providing medical services.Meanwhile,they are also critical for un-dertaking missions to handle health emergencies.This paper analyzes the necessity,current situation,and existing weaknesses of the health emergency capacity of public hospitals.It also conducts a comparative study of the emergency response mechanisms of foreign medical institutions.This paper aims to explore a well-developed design for health emergency in public hospitals that is suitable for China's medical conditions and to provide a feasible model for promoting high-quality health emergency management.
8.Clinical characteristics and management strategies of late bleeding after laparoscopic pancreaticoduodenectomy
Qiusheng LI ; Zejia ZHANG ; Zhongqiang XING ; Wei HE ; Weihong ZHAO ; Pengxiang LIU ; Ruibin LIU ; Jiansheng ZHANG ; Wenyan LU ; Jianhua LIU
Chinese Journal of Hepatobiliary Surgery 2023;29(8):589-593
Objective:To study the clinical characteristics and management strategies of late bleeding after laparoscopic pancreaticoduodenectomy (LPD).Methods:The clinical data of 58 patients with post-pancreaticoduodenectomy hemorrhage (PPH) admitted to the Department of Hepatobiliary Surgery of the Second Hospital of Hebei Medical University from March 2018 to March 2022 were retrospectively analyzed, including 42 males and 16 females, aged (61.88±11.02) years old. According to the occurrence of intra-abdominal erosion factors (e.g., pancreatic fistula, biliary fistula, gastrointestinal anastomotic fistula, intra-abdominal abscess), patients were divided into the erosion group ( n=42) and non-erosion group ( n=16). All patients underwent standard lymphadenectomy. Clinical data including the PPH time-point, occurrence of rebleeding, and treatment outcomes were accessed. The management strategies of PPH in the two groups of patients were analyzed. Results:The PPH time-point in the erosion group and non-erosion patients was 8.00 (5.00, 19.25) d and 21.50 (12.75, 26.75) d, respectively ( P=0.001). PPH can occurred within one month after surgery in both erosion and non-erosion groups. In the erosion group, 31 cases (73.81%, 31/42) were treated by re-operation, two (4.76%, 2/42) by interventional radiology and nine (21.43%, 9/42) with conservative protocol, respectively. In the non-erosion group, five cases (31.25%, 5/16) were treated by re-operation, seven (43.75%, 7/16) by interventional radiology and four (25.00%, 4/16) with conservative protocol, respectively. The incidence of re-bleeding is higher in the erosion group [47.6% (20/42) vs 12.5% (2/16), P<0.05]. Clinical manifestations, sites and severity of bleeding, and treatment outcomes were also different in the erosion and non-erosion groups (all P<0.05). Conclusions:The occurrence of intra-abdominal erosion factors can affect the clinical characteristics and treatment strategy of late bleeding after laparoscopic pancreaticoduodenectomy. Surgery remains the treatment of choice for post-pancreaticoduodenectomy hemorrhage either as an urgent or last resort.
9.FJX1 overexpression is associated with poor prognosis and promotes gastric cancer proliferation via the PI3K/AKT signaling pathway.
Hao ZHANG ; Zhen ZHANG ; Qiusheng WANG ; Lian WANG ; Zi YANG ; Zhijun GENG ; Yueyue WANG ; Jing LI ; Lugen ZUO
Journal of Southern Medical University 2023;43(6):975-984
OBJECTIVE:
To investigate the expression of four-jointed box kinase 1 (FJX1) in gastric cancer (GC), its correlation with survival outcomes of the patients, and its role in GC progression.
METHODS:
The expression level of FJX1 in GC tissues and normal gastric mucosal tissues and its correlation with the survival outcomes of GC patients were analyzed using TCGA and GEO database GC cohort. Immunohistochemistry was used to detect FJX1 expression level in clinical specimens of GC tissue, and its correlations with the patients' clinicopathological parameters and prognosis were analyzed. Bioinformatic analysis was performed to identify the potential pathways of FJX1 in GC. The effects of FJX1 overexpression or FJX1 silencing on GC cell proliferation and expressions of proliferation-related proteins, PI3K, AKT, p-PI3K, and p-AKT were evaluated using CCK-8 assay and Western blotting. The effect of FJX1 overexpression on GC cell tumorigenicity was evaluated in nude mice.
RESULTS:
GC tissues showed significantly higher expressions of FJX1 mRNA and protein compared with normal gastric mucosa tissues (P < 0.05). The high expression of FJX1 was associated with poor prognosis of GC patients (P < 0.05) and served as an independent risk factor for poor survival outcomes in GC (P < 0.05). FJX1 was expressed mainly in the cytoplasm of GC cells in positive correlation with Ki67 expression (R=0.34, P < 0.05), and was correlated with CA199 levels, depth of tumor infiltration and lymph node metastasis of GC (P < 0.05). In the cell experiment, FJX1 level was shown to regulate the expressions of Ki67 and PCNA and GC cell proliferation (P < 0.05). Gene set enrichment analysis indicated that the PI3K/AKT pathway potentially mediated the effect of FJX1, which regulated the expressions of PI3K and AKT and their phosphorylated proteins. In nude mice, FJX1 overexpression in GC cells significantly promoted the growth of the transplanted tumors (P < 0.05).
CONCLUSION
FJX1 is highly expressed in GC tissues and is correlated with poor prognosis of GC patients. FJX1 overexpression promotes GC cell proliferation through the PI3K/AKT signaling pathway, and may serve as a potential prognostic biomarker and therapeutic target for GC.
Animals
;
Mice
;
Cell Proliferation
;
Ki-67 Antigen
;
Mice, Nude
;
Phosphatidylinositol 3-Kinases
;
Proto-Oncogene Proteins c-akt
;
Signal Transduction
;
Stomach Neoplasms/pathology*
;
Humans
;
Intercellular Signaling Peptides and Proteins/genetics*
10.Norcantharidin induces apoptosis through autophagosome accumulation in breast cancer MDA-MB-231 cells
Yuan XIA ; Qingling JIANG ; Xiaoting WANG ; Minjing LI ; Qiusheng ZHENG ; Defang LI
Journal of China Pharmaceutical University 2023;54(6):757-768
The aim of this study was to investigate the effect of norcantharidin (NCTD) on the proliferation and apoptosis of triple-negative breast cancer cell line MDA-MB-231.Western blot was used to detect the effect of NCTD on the expression levels of apoptosis-related proteins Bax/Bcl-2, cleaved-PARP/PARP/PARP, cleved-caspase-9, cleaved-caspase-3 and MCL-1 in MDA-MB-231 cells.Also, the expression levels of autophagy-related proteins LC3-II/LC3-I, Parkin and PINK1 in MDA-MB-231 cells were measured by Western blot.Flow cytometry was used to measure the effect of NCTD on the changes of mitochondrial membrane potential and mitochondrial reactive oxygen species (ROS).The effect of NCTD on autophagy flow in cells expressing mCherry-EGFP-LC3 was detected by a confocal microscope.Moreover, the effects of NCTD combined with chloroquine (CQ) or 3-methyladenine (3-MA) on the apoptosis of MDA-MB-231 cells were detected by flow cytometry.The results showed that NCTD significantly increased the expression levels of Bax/Bcl-2, cleaved-PARP/PARP, cleaved-caspase-9, cleasved-caspase-3 and LC3-II/LC3-I proteins, and promoted the mitochondrial translocation of Parkin, and blocked the autophagic flow in MDA-MB-231 cells. Moreover, NCTD combined with CQ accelerated apoptosis, while NCTD combined with 3-MA decreased apoptosis.These results suggest that NCTD can induce autophagy accumulation and lead to apoptosis of MDA-MB-231 cells.

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