1.Construction of an ultrasound dynamic image segmentation model for thyroid nodules
Junpu HU ; Jialu LI ; Mengjie DOU ; Gang WANG ; Keyan LI ; Xiaofang FU ; Hao SUN ; Changqin SUN ; Duo SHI ; Yan LIAO ; Qiong WANG ; Faqin LYU
Chinese Journal of Ultrasonography 2025;34(6):518-524
Objective:To construct a thyroid nodule segmentation model using ultrasound dynamic images and explore its potential for assisting in the screening of thyroid nodules.Methods:A total of 126 patients with thyroid nodules(comprising 150 nodules)who were diagnosed and treated at Xuzhou Cancer Hospital from April 2024 to December 2024 were prospectively enrolled. Two-dimensional ultrasound was performed to capture short-axis and long-axis video images of thyroid nodules,forming a dynamic ultrasound image dataset. The dataset was divided into training,validation,and test sets in a ratio of 6∶1∶3. After the training loss curve converged,the model that performed well on the validation set was selected for testing. Three-fold cross-validation was employed for training and testing. All 300 ultrasound videos were divided into three subsets. In each experiment,two subsets were used as the training set,and one subset was used as the test set to evaluate the model's generalization ability. A collaborative spatiotemporal diffusion model was established based on the dynamic trends and tissue texture details of thyroid nodules. Six widely used segmentation metrics were employed to evaluate the model's application capabilities.Results:The study included 126 patients with 150 thyroid nodules,300 dynamic ultrasound images,and video lengths of 3-4 seconds per nodule,resulting in 12 312 segmented images. The size of the thyroid nodules was(10.7 ± 10.6)mm(transverse diameter)×(8.4 ± 6.3)mm(anteroposterior diameter). Among the nodules,62(41.3%)had clear boundaries,while 88(58.7%)had indistinct boundaries;61(40.7%)exhibited regular shapes,while 89(59.3%)were irregular;66(44.0%)had a taller-than-wide aspect ratio;and 70(46.7%)showed microcalcifications. The collaborative diffusion model based on dynamic ultrasound image segmentation achieved the following scores:a Jaccard score of(69.22 ± 0.03)%,a Dice score of(79.16 ± 0.18)%,a Precision score of(86.70 ± 0.17)%,a Recall score of(77.82 ± 0.04)%,an Sα score of(85.26 ± 0.01)%,and an Eθmn score of(90.58 ± 0.17)%. Compared to other models,this model demonstrated significant improvements across all evaluation metrics,achieving the highest values in each metric with increments of over 8% and 1%,respectively. Conclusions:The collaborative diffusion model with a dynamic controller,constructed based on dynamic ultrasound images of thyroid nodules,demonstrates excellent performance in ultrasound image segmentation. It improves the accuracy of thyroid nodule screening,thereby providing a valuable auxiliary diagnostic tool for clinical practice.
2.Automatic recognition and segmentation of brachial plexus in ultrasonic images based on deep learning
Duo SHI ; Han ZHANG ; Peipei LIU ; Ruichao ZHANG ; Qingyu LIU ; Hao SUN ; Xiaofang FU ; Mengjie DOU ; Junpu HU ; Changqin SUN ; Keyan LI ; Jianqiu HU ; Guangquan ZHOU ; Ligang CUI ; Ping ZHOU ; Faqin LYU
Chinese Journal of Ultrasonography 2025;34(9):737-744
Objective:To propose a deep learning(DL)-based ultrasound imaging auxiliary tool for automatic segmentation and recognition of the brachial plexus(BP),and to enhance the accuracy and safety of clinical procedures.Methods:It was a multicenter study that collected 773 healthy subjects from Peking University Third Hospital and its branch campuses,the Third Medical Center of the Chinese PLA General Hospital,and Shanghai Eighth People's Hospital between August 2024 and February 2025. Brachial plexus(BP)images in the interscalene groove were captured used high-frequency ultrasound by senior sonographers,a dataset comprising 1 289 standardized images were constructed and the improved model(CHA-TransUNet)was trained. The test set was input into 6 different models(CHA-TransUNet,R50-Unet,TransUnet,SegFormer,SwinUnet,MISSFormer)for segmentation. Segmentation accuracy was evaluated using metrics including the Dice similarity coefficient(DSC),95% Hausdorff distance(HD95)and mean intersection over union(mIoU),and was compared with the segmentation results of 3 ultrasound physicians with varying experience levels(junior physicians and senior physicians)to validate the model's segmentation efficacy.Results:The CHA-TransUNet model established based on a dataset of 1 289 standardized images achieved segmentation results for the BP with a DSC of 90.15%,mIoU of 91.02%,and HD95 of 8.08. Its accuracy was higher than other mainstream models(DSC:90.15% vs. 87.60%,87.77%,81.35%,84.78%,84.55%),significantly better than junior physicians(DSC:90.15% vs. 68.73%, Z=-127.76, P<0.001),and approached the level of senior physician(DSC:90.15% vs. 86.15%, Z=-31.33, P=0.549). The model demonstrated superior boundary recognition in complex anatomical structures(e.g.,C6/C7 nerve roots)compared to ultrasound physicians(junior and senior)(HD95:8.08 vs. 26.34,17.44,56.80). Conclusions:This study proposes an analysis model for BP ultrasound images,CHA-TransUNet. This model achieves segmentation and recognition of the BP with relatively complex pathways and structures. The model exhibits high accuracy and stability,outperforming current mainstream network models and junior physicians while approaching the performance level of senior physicians. It assists junior physicians or trainees in more accurately identifying and localizing the BP.
3.The role and mechanism of ERK-mediated Drp1 signaling in exercise-induced skeletal muscle damage
Mengyu LI ; Hao DENG ; Shiqiao ZHENG ; Duo ZHANG ; Tianai YANG ; Ranggui MA ; Zhi XIA ; Huayu SHANG
Chinese Journal of Sports Medicine 2025;44(9):717-728
Objective To observe the effect of a heavy load exercise on the ultrastructure,function and fission of skeletal muscle mitochondria in rats,and to analyze the changes of the phosphorylation expression of mitochondrial fission protein and upstream kinase at different times postexercise,and to explore the effect of acute heavy load exercise on mitochondrial fission in skeletal muscle of rats and its possible mechanism.Methods Forty-eight adult male Sprague-Dawley rats were randomly divided in-to a quiet control group(C,n=8)and an exercise group(E,n=40).Rats in the E group exercised on a treadmill down a 16° decline at 16 m/min for 90 min and were further divided into 0 h,12 h,24 h,48 h,and 72 h postexercise subgroups.Soleus muscle was isolated and mitochondria were ex-tracted at the corresponding time points after exercise.The ultrastructure of mitochondria in the soleus muscle was observed using transmission electron microscopy,and mitochondrial quantity and morphomet-ric analysis were conducted.Moreover,the colocalization and quantity of dynamin-related protein 1(Drp1)and cytochrome C oxidease subunit Ⅳ(COXⅣ)in the soleus muscle were detected using im-munofluorescence double-labeling techniques.Meanwhile,protein levels of soleus musclep-Drp1Ser616,p-Drp1Ser637,p-extracellular regulatory protein kinaseThr202/Tyr204(p-ERKThr202/Tyr204),p-protein kinaseAThr197(p-PKAThr197),and mitochondrial NADH of ubiquinone oxidoreductase subunit B8(NDUFB8)and ubiqui-nol-cytochrome C reductase core protein 2(UQCRC2)were determined by using Western blotting.An-other twenty-four rats were randomly divided into a DMSO group(CD),a U0126 group(CU),an Ex-ercise+DMSO group(ED),and an Exercise+U0126 group(EU).Six mice in each group were giv-en a single intra-bitoneal injection of DMSO or ERK inhibitor U0126 20 min before acute downhill running.Then,their phosphorylated expressions of ERKThr202/Tyr204 and Drp1Ser616 in soleus muscle were de-tected by Western blotting.Results(1)From 0 h to 48 h after exercise,the soleus muscle mitochon-dria showed swelling,rounding,and uneven distribution of mitochondria,among which the degree of mitochondrial damage was the most serious at 12 h and 24 h after exercise.Moreover,the protein ex-pression of NDUFB8 and UQCRC2 in the mitochondria fractions from soleus muscle was significantly lower at 12 h post-exercise(P<0.05).(2)The co-localization of Drp1 and COXⅣ in the skeletal muscle increased significantly at 12 h to 24 h after a heavy load exercise compared with group C and group E0(P<0.01).Moreover,the mitochondrial area,circumference,aspect ratio and Ferret diameter in the skeletal muscle were significantly lower at 12 h to 24 h postexercise(P<0.05).Meanwhile,the number of mitochondria was significantly higher at 24 h after exercise(P<0.01).(3)The phosphoryla-tion of ERKThr202/Tyr204,PKAThr197 and Drp1Ser616 was significantly higher at 24 h after exercise(P<0.01),while that of Drp1Ser637 was significantly lower at 48 h and 72 h post-exercise(P<0.01).However,the phosphorylated expressions of ERKThr202/Tyr204 and Drp1Ser616 were significantly down-regulated by U0126 treatment before exercise.Conclusion A session of heavy load exercise caused mitochondrial structure and function damage and induced mitochondrial fission in the skeletal muscle,and then to maintain the homeostasis of skeletal muscle cells by cleaving damaged mitochondria.The mechanism of promot-ing skeletal muscle repair may be related to the positive and negative regulation of Drp1 activity by the phosphorylation of Drp1Ser616 and Drp1Ser637,respectively.Among them,the activation of ERKThr202/Tyr204 mediates the phosphorylation activation of Drp1Ser616,but PKAThr197 is not an upstream kinase that medi-ates the inactivation of Drp1Ser637 phosphorylation.
4.Screening of pyroptosis-related genes in pulmonary plague and molecular mechanisms based on bioinformatics analysis
Hao XIE ; Lingli AI ; Duo SU ; Lu LI ; Lingfei HU ; Dongsheng ZHOU ; Bo GAO ; Huiying YANG
Military Medical Sciences 2025;49(4):281-288
Objective To explore the biological functions of pyroptosis-related genes in pneumonic plague using bioinformatics methods,and to evaluate their potential applicability as diagnostic markers.Methods The pneumonic plague-related dataset GSE220123 was retrieved from the Gene Expression Omnibus(GEO)database and screened for differentially expressed pyroptosis-related genes(DE-PRGs).The functions of DE-PRGs were studied via Gene Ontology(GO),Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses,and immune infiltration analysis.The hub genes were identified via protein-protein interaction(PPI)network analysis,and further screened for key genes with sustained high expression characteristics based on differential expression analysis.The relative expression levels of the key genes were verified using the reverse transcription real-time quantitative PCR(qPCR)method.Results A total of 17 DE-PRGs were screened,and PPI network analysis revealed 7 Hub genes.Among them,Casp4 continued to be up-regulated during the course of pneumonic plague.The results of reverse transcription qPCR were consistent with the those of bioinformatic analyses.Conclusion DE-PRGs play a crucial role in the immune response of pneumonic plague,especially Casp4,which has significant applications as a diagnostic biomarker and potential therapeutic target for pneumonic plague.
5.Preliminary efficacy observation of 3D printed functional spinal external fixation brace combined with McKenzie therapy in the treatment of lumbar disc herniation.
Ning-Xia WANG ; Ping CHEN ; Hai-Dong WANG ; Jing JI ; Fang-Hong NIAN ; Xin LIU ; Chong-Fei JIN ; Duo-Ming ZHAO ; Hao-Lin LI ; Wei-Gang CHENG ; Gui-Lin LAI ; Guo-Biao WU
China Journal of Orthopaedics and Traumatology 2025;38(10):1047-1054
OBJECTIVE:
To observe the clinical efficacy of 3D printing spinal external fixator combined with McKenzie therapy for patients with lumbar dics herniation (LDH).
METHODS:
Sixty patients with LDH between January 2022 and January 2023 were enrolled. Among them, 30 patients were given McKinsey training. According to different treatment methods, all patients were divided into McKenzie group and McKenzie + 3D printing group, 30 patients in each group. The McKenzie group provided McKenzie therapy. The McKenzie + 3D printing group were treated with 3D printing spinal external fixation brace on the basis of McKenzie therapy. Patients in both groups were between 25 and 60 years of age and had their first illness. In the McKenzie group, there were 19 males and 11 females, with an average age of (48.57±5.86) years old, and the disease duration was (7.03 ±2.39) months. The McKenzie + 3D printing group, there were 21 males and 9 females, with an average age of (48.80±5.92) years old, and the disease duration was(7.30±2.56) months. Pain was evaluated using the visual analogue scale (VAS), and lumbar spine function was assessed using the Oswestry disability index (ODI) and the Japanese Orthopaedic Association (JOA) score. VAS, ODI and JOA scores were compared between two groups before treatment and at 1, 3, 6, 9 and 12 months after treatment.
RESULTS:
All patients were followed up for 12 months. The VAS for the McKenzie combined with 3D printing group before treatment and at 1, 3, 6, 9, and 12 months post-treatment were(6.533±0.860), (5.133±1.008), (3.933±0.868), (2.900±0.759), (2.067±0.640), (1.433±0.504), respectively. In the McKenzie group, the corresponding scores were (6.467±0.860), (5.067±1.048), (4.600±0.968), (3.533±1.008), (2.567±0.728), (1.967±0.809), respectively. The ODI of the McKenzie group before treatment and at 1, 3, 6, 9, and 12 months post-treatment were (41.033±6.810)%, (37.933±6.209)%, (35.467±6.962)%, (27.567±10.081)%, (20.800±7.531)%, (13.533±5.158)%, respectively. For the McKenzie combined with 3D printing group, the corresponding ODI were(38.033±5.605)%, (33.000±6.192)%, (28.767±7.045)%, (22.200±5.517)%, (17.700±4.836)%, (11.900±2.771)%, respectively. The JOA scores of the McKenzie combined with 3D printing group before treatment and at 1, 3, 6, 9, and 12 months post-treatment were(8.900±2.074), (13.133±2.330), (15.700±3.583), (20.400±3.480), (22.267±3.084), (24.833±2.640), respectively. In the McKenzie group, the corresponding scores were(9.200±2.091), (12.267±2.406), (15.333±3.198), (18.467±2.240), (20.133±2.751), (22.467±2.849), respectively. Before the initiation of treatment, no statistically significant differences were observed in the VAS, ODI, and JOA scores between two groups (P>0.05). At 3, 6, 9, and 12 months post-treatment, the VAS in the McKenzie combined with 3D printing group was significantly lower than that in the McKenzie group, and the difference was statistically significant (P<0.05). The comparison of ODI between two groups at 1, 3, 6, 9, and 12 months post-treatment revealed statistically significant differences (P<0.05). At 6, 9, and 12 months post-treatment, the JOA score in the McKenzie combined with 3D printing group was significantly higher than that in the McKenzie-only group, and the difference was statistically significant (P<0.05).
CONCLUSION
The combination of 3D printed functional spinal external fixation brace with McKenzie therapy can significantly improve and maintain lumbar function in patients with LDH.
Humans
;
Male
;
Female
;
Middle Aged
;
Printing, Three-Dimensional
;
Intervertebral Disc Displacement/surgery*
;
External Fixators
;
Lumbar Vertebrae/surgery*
;
Adult
;
Braces
;
Treatment Outcome
6.The role and mechanism of ERK-mediated Drp1 signaling in exercise-induced skeletal muscle damage
Mengyu LI ; Hao DENG ; Shiqiao ZHENG ; Duo ZHANG ; Tianai YANG ; Ranggui MA ; Zhi XIA ; Huayu SHANG
Chinese Journal of Sports Medicine 2025;44(9):717-728
Objective To observe the effect of a heavy load exercise on the ultrastructure,function and fission of skeletal muscle mitochondria in rats,and to analyze the changes of the phosphorylation expression of mitochondrial fission protein and upstream kinase at different times postexercise,and to explore the effect of acute heavy load exercise on mitochondrial fission in skeletal muscle of rats and its possible mechanism.Methods Forty-eight adult male Sprague-Dawley rats were randomly divided in-to a quiet control group(C,n=8)and an exercise group(E,n=40).Rats in the E group exercised on a treadmill down a 16° decline at 16 m/min for 90 min and were further divided into 0 h,12 h,24 h,48 h,and 72 h postexercise subgroups.Soleus muscle was isolated and mitochondria were ex-tracted at the corresponding time points after exercise.The ultrastructure of mitochondria in the soleus muscle was observed using transmission electron microscopy,and mitochondrial quantity and morphomet-ric analysis were conducted.Moreover,the colocalization and quantity of dynamin-related protein 1(Drp1)and cytochrome C oxidease subunit Ⅳ(COXⅣ)in the soleus muscle were detected using im-munofluorescence double-labeling techniques.Meanwhile,protein levels of soleus musclep-Drp1Ser616,p-Drp1Ser637,p-extracellular regulatory protein kinaseThr202/Tyr204(p-ERKThr202/Tyr204),p-protein kinaseAThr197(p-PKAThr197),and mitochondrial NADH of ubiquinone oxidoreductase subunit B8(NDUFB8)and ubiqui-nol-cytochrome C reductase core protein 2(UQCRC2)were determined by using Western blotting.An-other twenty-four rats were randomly divided into a DMSO group(CD),a U0126 group(CU),an Ex-ercise+DMSO group(ED),and an Exercise+U0126 group(EU).Six mice in each group were giv-en a single intra-bitoneal injection of DMSO or ERK inhibitor U0126 20 min before acute downhill running.Then,their phosphorylated expressions of ERKThr202/Tyr204 and Drp1Ser616 in soleus muscle were de-tected by Western blotting.Results(1)From 0 h to 48 h after exercise,the soleus muscle mitochon-dria showed swelling,rounding,and uneven distribution of mitochondria,among which the degree of mitochondrial damage was the most serious at 12 h and 24 h after exercise.Moreover,the protein ex-pression of NDUFB8 and UQCRC2 in the mitochondria fractions from soleus muscle was significantly lower at 12 h post-exercise(P<0.05).(2)The co-localization of Drp1 and COXⅣ in the skeletal muscle increased significantly at 12 h to 24 h after a heavy load exercise compared with group C and group E0(P<0.01).Moreover,the mitochondrial area,circumference,aspect ratio and Ferret diameter in the skeletal muscle were significantly lower at 12 h to 24 h postexercise(P<0.05).Meanwhile,the number of mitochondria was significantly higher at 24 h after exercise(P<0.01).(3)The phosphoryla-tion of ERKThr202/Tyr204,PKAThr197 and Drp1Ser616 was significantly higher at 24 h after exercise(P<0.01),while that of Drp1Ser637 was significantly lower at 48 h and 72 h post-exercise(P<0.01).However,the phosphorylated expressions of ERKThr202/Tyr204 and Drp1Ser616 were significantly down-regulated by U0126 treatment before exercise.Conclusion A session of heavy load exercise caused mitochondrial structure and function damage and induced mitochondrial fission in the skeletal muscle,and then to maintain the homeostasis of skeletal muscle cells by cleaving damaged mitochondria.The mechanism of promot-ing skeletal muscle repair may be related to the positive and negative regulation of Drp1 activity by the phosphorylation of Drp1Ser616 and Drp1Ser637,respectively.Among them,the activation of ERKThr202/Tyr204 mediates the phosphorylation activation of Drp1Ser616,but PKAThr197 is not an upstream kinase that medi-ates the inactivation of Drp1Ser637 phosphorylation.
7.Construction of an ultrasound dynamic image segmentation model for thyroid nodules
Junpu HU ; Jialu LI ; Mengjie DOU ; Gang WANG ; Keyan LI ; Xiaofang FU ; Hao SUN ; Changqin SUN ; Duo SHI ; Yan LIAO ; Qiong WANG ; Faqin LYU
Chinese Journal of Ultrasonography 2025;34(6):518-524
Objective:To construct a thyroid nodule segmentation model using ultrasound dynamic images and explore its potential for assisting in the screening of thyroid nodules.Methods:A total of 126 patients with thyroid nodules(comprising 150 nodules)who were diagnosed and treated at Xuzhou Cancer Hospital from April 2024 to December 2024 were prospectively enrolled. Two-dimensional ultrasound was performed to capture short-axis and long-axis video images of thyroid nodules,forming a dynamic ultrasound image dataset. The dataset was divided into training,validation,and test sets in a ratio of 6∶1∶3. After the training loss curve converged,the model that performed well on the validation set was selected for testing. Three-fold cross-validation was employed for training and testing. All 300 ultrasound videos were divided into three subsets. In each experiment,two subsets were used as the training set,and one subset was used as the test set to evaluate the model's generalization ability. A collaborative spatiotemporal diffusion model was established based on the dynamic trends and tissue texture details of thyroid nodules. Six widely used segmentation metrics were employed to evaluate the model's application capabilities.Results:The study included 126 patients with 150 thyroid nodules,300 dynamic ultrasound images,and video lengths of 3-4 seconds per nodule,resulting in 12 312 segmented images. The size of the thyroid nodules was(10.7 ± 10.6)mm(transverse diameter)×(8.4 ± 6.3)mm(anteroposterior diameter). Among the nodules,62(41.3%)had clear boundaries,while 88(58.7%)had indistinct boundaries;61(40.7%)exhibited regular shapes,while 89(59.3%)were irregular;66(44.0%)had a taller-than-wide aspect ratio;and 70(46.7%)showed microcalcifications. The collaborative diffusion model based on dynamic ultrasound image segmentation achieved the following scores:a Jaccard score of(69.22 ± 0.03)%,a Dice score of(79.16 ± 0.18)%,a Precision score of(86.70 ± 0.17)%,a Recall score of(77.82 ± 0.04)%,an Sα score of(85.26 ± 0.01)%,and an Eθmn score of(90.58 ± 0.17)%. Compared to other models,this model demonstrated significant improvements across all evaluation metrics,achieving the highest values in each metric with increments of over 8% and 1%,respectively. Conclusions:The collaborative diffusion model with a dynamic controller,constructed based on dynamic ultrasound images of thyroid nodules,demonstrates excellent performance in ultrasound image segmentation. It improves the accuracy of thyroid nodule screening,thereby providing a valuable auxiliary diagnostic tool for clinical practice.
8.Automatic recognition and segmentation of brachial plexus in ultrasonic images based on deep learning
Duo SHI ; Han ZHANG ; Peipei LIU ; Ruichao ZHANG ; Qingyu LIU ; Hao SUN ; Xiaofang FU ; Mengjie DOU ; Junpu HU ; Changqin SUN ; Keyan LI ; Jianqiu HU ; Guangquan ZHOU ; Ligang CUI ; Ping ZHOU ; Faqin LYU
Chinese Journal of Ultrasonography 2025;34(9):737-744
Objective:To propose a deep learning(DL)-based ultrasound imaging auxiliary tool for automatic segmentation and recognition of the brachial plexus(BP),and to enhance the accuracy and safety of clinical procedures.Methods:It was a multicenter study that collected 773 healthy subjects from Peking University Third Hospital and its branch campuses,the Third Medical Center of the Chinese PLA General Hospital,and Shanghai Eighth People's Hospital between August 2024 and February 2025. Brachial plexus(BP)images in the interscalene groove were captured used high-frequency ultrasound by senior sonographers,a dataset comprising 1 289 standardized images were constructed and the improved model(CHA-TransUNet)was trained. The test set was input into 6 different models(CHA-TransUNet,R50-Unet,TransUnet,SegFormer,SwinUnet,MISSFormer)for segmentation. Segmentation accuracy was evaluated using metrics including the Dice similarity coefficient(DSC),95% Hausdorff distance(HD95)and mean intersection over union(mIoU),and was compared with the segmentation results of 3 ultrasound physicians with varying experience levels(junior physicians and senior physicians)to validate the model's segmentation efficacy.Results:The CHA-TransUNet model established based on a dataset of 1 289 standardized images achieved segmentation results for the BP with a DSC of 90.15%,mIoU of 91.02%,and HD95 of 8.08. Its accuracy was higher than other mainstream models(DSC:90.15% vs. 87.60%,87.77%,81.35%,84.78%,84.55%),significantly better than junior physicians(DSC:90.15% vs. 68.73%, Z=-127.76, P<0.001),and approached the level of senior physician(DSC:90.15% vs. 86.15%, Z=-31.33, P=0.549). The model demonstrated superior boundary recognition in complex anatomical structures(e.g.,C6/C7 nerve roots)compared to ultrasound physicians(junior and senior)(HD95:8.08 vs. 26.34,17.44,56.80). Conclusions:This study proposes an analysis model for BP ultrasound images,CHA-TransUNet. This model achieves segmentation and recognition of the BP with relatively complex pathways and structures. The model exhibits high accuracy and stability,outperforming current mainstream network models and junior physicians while approaching the performance level of senior physicians. It assists junior physicians or trainees in more accurately identifying and localizing the BP.
9.Genetic analysis and assisted reproductive guidance for two infertile patients with rare small supernumerary marker chromosomes
Duo YI ; Shimin YUAN ; Liang HU ; Fei GONG ; Keli LUO ; Hao HU ; Yueqiu TAN ; Guangxiu LU ; Ge LIN ; Dehua CHENG
Chinese Journal of Medical Genetics 2024;41(5):519-525
Objective:To carry out cytogenetic and molecular genetic analysis for two infertile patients carrying rare small supernumerary marker chromosomes (sSMC).Methods:Two infertile patients who received reproductive and genetic counseling at CITIC Xiangya Reproductive and Genetic Hospital on October 31, 2018 and May 10, 2021, respectively were selected as the study subjects. The origin of sSMCs was determined by conventional G banding, fluorescence in situ hybridization (FISH) and copy number variation sequencing (CNV-seq). Microdissection combined with high-throughput whole genome sequencing (MicroSeq) was carried out to determine the fragment size and genomic information of their sSMCs. Results:For patient 1, G-banded karyotyping and FISH revealed that he has a karyotype of mos47, XY, del(16)(p10p12), + mar[65]/46, XY, del(16)(p10p12)[6]/48, XY, del(16)(p10p12), + 2mar[3].ish mar(Tel 16p-, Tel 16q-, CEP 16-, WCP 16+ ). CNV analysis has yielded a result of arr[GRCh37]16p12.1p11.2(24999364_33597595)×1[0.25]. MicroSeq revealed that his sSMC has contained the region of chromosome 16 between 24979733 and 34023115 (GRCh37). For patient 2, karyotyping and reverse FISH revealed that she has a karyotype of mos 47, XX, + mar[37]/46, XX[23].rev ish CEN5, and CNV analysis has yielded a result of seq[GRCh37]dup(5)(p12q11.2)chr5: g(45120001_56000000)dup[0.8]. MicroSeq results revealed that her sSMC has contained the region of chromosome 5 between 45132364 and 55967870(GRCh37). After genetic counseling, both couples had opted in vitro fertilization (IVF) treatment and preimplantation genetic testing (PGT). Conclusion:For individuals harboring sSMCs, it is vital to delineate the origin and structural characteristics of the sSMCs for their genetic counseling and reproductive guidance. Preimplantation genetic testing after microdissection combined with high-throughput whole genome sequencing (MicroSeq-PGT) can provide an alternative treatment for carrier couples with a high genetic risk.
10.Role of FKBP8-mediated mitophagy in repetitive eccentric exercise-induced skeletal muscle damage
Shiqiao ZHENG ; Hao DENG ; Mengyu LI ; Duo ZHANG ; Tianai YANG ; Ranggui MA ; Zhi XIA ; Huayu SHANG
Chinese Journal of Sports Medicine 2024;43(6):473-483
Objective To explore the effect of 4-week repetitive downhill treadmill running on the mi-tochondrial structure,function,and autophagy in skeletal muscle of rats,so as to analyze the role of FKBP8-mediated mitophagy in exercise-induced mitochondrial damage in their skeletal muscles.Meth-ods Thirty-two male adult Sprague-Dawley rats were randomly divided into a 2-week quiet control group(2C group,n=8),a 4-week quiet control group(4C group,n=8),a 2-week exercise group(2E group,n=8)and a 4-week exercise group(4E group,n=8).Rats in 2E and 4E groups performed dai-ly 90-minute downhill treadmill running(-16°,16 m/min)5 days a week for two and four weeks,re-spectively.Then,they rested for 24 hours and received an exhaustive exercise test.Running distance and blood lactate were measured prior to and at the time of exercise cessation.Moreover,mitochondri-al ultrastructural changes in soleus muscles were observed by using a transmission electron microscope.The protein expression of mitochondrial succinate dehydrogenase subunit B(SDHB),cytochrome C oxi-dase subunit 1(MTCO1),FK506 binding protein 8(FKBP8)and microtubule associated protein 1 light chain 3(LC3)in the soleus muscle were measured using Western blotting.Meanwhile,the co-localiza-tion of FKBP8 with LC3 and cytochrome C oxidase subunit Ⅳ(COXⅣ)with LC3,lysosomal associat-ed membrane protein 2(LAMP2)were detected by the immunofluorescence double labeling technique.Results(1)The running distance of one exhaustive exercise and the blood lactate before and after the test in 2E group were significantly higher than 2C and 4E groups(P<0.05 or P<0.01),and the run-ning distance of 4E group was significantly higher than 4C group(P<0.01).However,there was no sig-nificant difference between 4E and 4C groups in the blood lactate before and after the exhaustive exer-cise test(P>0.05).(2)In both 2E and 4E groups,significant mitochondrial swelling and accumulation under cell membrane,as well as a number of mitophagosomes and mitophagolysosomes were observed,together with a significant reduce in the number of mitochondria(P<0.05),which was more severe in 2E group than 4E group.(3)The protein expression of mitochondrial SDHB and MTCO1 in 2E and 4E groups were lower than 2C and 4C groups,respectively,with significantly greater changes of these proteins in 4E group than 2E group(P<0.05 or P<0.01).(4)The protein expression of mitochondrial FKBP8 and LC3,as well as the co-localization of FKBP8 with LC3 and COXⅣ with LC3,LAMP2 in 2E and 4E groups were higher than 2C and 4C groups,respectively,with significantly greater changes in 4E group than 2E group(P<0.05 or P<0.01).Conclusion After 4-week downhill treadmill running,the structure,quantity and function of mitochondria in skeletal muscle are impaired.FKBP8-mediated mitophagy is activated,but is insufficient to degrade the damaged mitochondria,leading to muscular damage,as well as the increasing and falling down of running capacity.

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