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.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.
3.Effect of Glycyrrhizae Radix et Rhizoma-containing Serum on LPS-induced Inflammation in Caco2 Cells Based on Inhibition of Ferroptosis by Nrf2/HO-1 Pathway
Jinrong KONG ; Gaoxiang SHI ; Jing HOU ; Ye FENG ; Qingzhen XIANG ; Yunlai WANG ; Zihua XUAN ; Fan XU
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(16):144-153
ObjectiveTo investigate the effect of Glycyrrhizae Radix et Rhizoma (GR)-containing serum on lipopolysaccharide (LPS)-induced inflammation in human colon epithelial adenocarcinoma cells (Caco2) based on inhibition of ferroptosis by the nuclear factor erythroid 2-related factor 2 (Nrf2)/heme oxygenase-1 (HO-1) pathway. MethodCaco2 cells were divided into a normal group, a model group (LPS, 200 μg·L-1), low-, medium-, and high-dose GR-containing serum groups (5%, 10%, 20%), and a ferroptosis inhibitor group (3-amino-4-cyclohexylamino-benzoic acid ethyl ester, Fer-1, 10 μmol·L-1). The cells in the normal group were cultured normally, while those in other groups underwent the induction of an inflammation model. The cells in the low-, medium-, and high-dose GR-containing serum groups were treated with 5%, 10%, and 20% GR-containing serum for 24 hours, respectively, and the cells in the ferroptosis inhibitor group were treated with Fer-1 for 24 hours. Transmission electron microscopy was used to observe mitochondrial morphology in each group. Flow cytometry was used to detect intracellular Fe2+ levels. Microplate assays were performed to measure superoxide dismutase (SOD) activity, malondialdehyde (MDA) and glutathione peroxidase (GSH-Px) levels. Enzyme-linked immunosorbent assay (ELISA) was used to measure interleukin-1β (IL-1β), IL-6, IL-10, and tumor necrosis factor-α (TNF-α) levels. Western blot was used to measure the expression levels of Nrf2, HO-1, ferritin heavy chain 1 (FTH1), and glutathione peroxidase 4 (GSH-Px4) proteins. Small interfering RNA (siRNA) was used to investigate the role of Nrf2 in ferroptosis regulation. The cells after interference were divided into a negative control (NC) group, a Si-Nrf2 group, a GR-containing serum (20%) + Si-Nrf2 group, and a GR-containing serum (20%) + NC group. Microplate assays were performed to measure MDA, SOD, and GSH-Px levels, and Western blot was used to measure the expression levels of Nrf2, HO-1, FTH1, and GSH-Px4 proteins. ResultCompared with the normal group, the model group showed mitochondrial contraction, increased mitochondrial membrane thickness, and smaller mitochondrial morphology, increased Fe2+ content (P<0.01), blunted SOD activity (P<0.01), decreased GSH-Px expression (P<0.01), increased MDA content (P<0.01), reduced expression levels of Nrf2 and HO-1 (P<0.05), reduced FTH1 expression (P<0.01), and down-regulated GSH-Px4 expression (P<0.01). In the GR-containing serum groups, the medium- and high-dose groups showed a significant decrease in Fe2+ content (P<0.01), potentiated SOD and GSH-Px activities (P<0.01), and decreased MDA levels (P<0.01). The high-dose group showed a significant increase in Nrf2 expression (P<0.05), and the medium-dose group showed increased expression of HO-1 and GSH-Px4 proteins (P<0.05). The expression levels of FTH1 significantly increased in the low-, medium-, and high-dose groups (P<0.01). The study on mechanism revealed that compared with the NC group, the cells transfected with Nrf2 siRNA showed increased MDA content (P<0.01), blunted SOD activity (P<0.01), decreased GSH-Px activity (P<0.01), decreased expression of Nrf2 and HO-1 (P<0.01), and reduced levels of FTH1 and GSH-Px4 proteins (P<0.01). Compared with the Si-Nrf2 group, the cells treated with GR-containing serum showed a decrease in MDA content (P<0.01), an increase in SOD activity (P<0.01), an increase in GSH-Px activity (P<0.01), increased expression of Nrf2 and FTH1 proteins (P<0.05), and higher expression levels of HO-1 and GSH-Px4 proteins (P<0.01). ConclusionGR-containing serum can reduce the inflammatory cytokines and oxidative stress levels in LPS-induced Caco2 cells. Its mechanism is related to the promotion of Nrf2/HO-1 signaling pathway expression, alleviating intracellular lipid peroxidation and inhibiting ferroptosis.

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