1.Virtual cutting-based morphological differences in osteoarthritic and healthy knees: Implications for total knee arthroplasty prosthesis design.
Bin YU ; Yu ZHANG ; Dongdong CAO ; Jinchang HAN ; Weiyong WU ; Chao ZHANG ; Aifeng LIU
Chinese Journal of Traumatology 2025;28(6):436-444
PURPOSE:
End-stage knee osteoarthritis (OA) patients are the primary candidates for total knee arthroplasty (TKA). However, most morphological refinements of TKA prosthesis are based on anatomical data from the knees of healthy individuals. This study aimed to determine whether differences exist in key bony morphological characteristics of the distal femur and proximal tibia between osteoarthritic knees and healthy knees.
METHODS:
This was a retrospective cross-sectional observational study with a case-control design. Patients who were aged ≥ 50 years, had no history of trauma, fracture, or surgery in the studied knee, and had no obvious knee flexion contracture were included in this study by CT scans. Patients who met the American College of Rheumatology clinical criteria for knee OA were included in the study group. Kellgren-Lawrence grade III or IV knees were studied (for bilateral cases, the more severely affected knee was chosen). Patients who presented with unilateral knee pain or trauma were included in the control group, with CT scans from the opposite (asymptomatic) knee used for analyzing. The studied knee had a Kellgren-Lawrence grade of 0 or I and showed no abnormalities upon physical examination. Archived knee CT scans from 160 patients were divided into 2 groups: the study group (80 moderate-to-severe OA knees) and the control group (80 healthy knees). After 3-dimensional reconstruction and virtual cutting using a CT workstation, 13 morphological parameters of the distal femur and proximal tibia were compared between the 2 groups using independent-samples t-tests.
RESULTS:
No significant group differences in the femoral anteroposterior dimension (p = 0.797), height of the lateral femoral condyle (p = 0.268), posterior condylar angle (p = 0.240), tibial anteroposterior dimension (p = 0.536), or tibial lateral anteroposterior dimension (p = 0.702) were observed. However, the femoral mediolateral dimension (p = 0.002), distal femoral aspect ratio (femoral mediolateral dimension/femoral anteroposterior dimension) (p < 0.001), height of the femoral trochlear groove (p < 0.001), height of the medial femoral condyle (p < 0.001), tibial mediolateral dimension (p = 0.001), proximal tibial aspect ratio (tibial mediolateral dimension/tibial anteroposterior dimension) (p = 0.004), tibial medial anteroposterior dimension (p = 0.005), and tibial asymmetry ratio (tibial medial anteroposterior dimension/tibial lateral anteroposterior dimension) (p = 0.006) were all significantly greater in the study group.
CONCLUSION
Knees with moderate-to-severe OA are significantly wider than healthy knees, and OA is a risk factor for increased tibial platform asymmetry. When refining the morphological parameters of TKA prostheses, the specific bony morphological characteristics of OA knees should be taken into account to reduce the potential risk of femoral or tibial component underhang and facilitate optimal balance between tibial component fit and rotational alignment.
Humans
;
Osteoarthritis, Knee/pathology*
;
Male
;
Female
;
Cross-Sectional Studies
;
Retrospective Studies
;
Arthroplasty, Replacement, Knee
;
Middle Aged
;
Aged
;
Case-Control Studies
;
Prosthesis Design
;
Knee Prosthesis
;
Femur/anatomy & histology*
;
Tibia/anatomy & histology*
;
Tomography, X-Ray Computed
;
Knee Joint/diagnostic imaging*
2.Choline theophyllinate promotes activation of primordial follicles in mice through PI3K/Akt signaling pathway
Tiantian HAO ; Wenbo ZHANG ; Weiyong WANG ; Meijia ZHANG
Chinese Journal of Pathophysiology 2025;41(6):1145-1152
AIM:To investigate the function and mechanism of the smooth muscle relaxant choline theophylli-nate(CT)in activating primordial follicles in mice.METHODS:Experiments were conducted using in vitro culture of 3-day postpartum(dpp)neonatal SPF-grade female mice,intraperitoneal injection in 3-dpp neonatal mice,and oral adminis-tration in 21-dpp adolescent female mice.The mice were divided into control and CT groups.The ovaries were isolated from 3-dpp neonatal mice for the in vitro culture.Hematoxylin staining was used to count the number of primordial and growing follicles,with 10 mice in each group.qPCR was performed to analyze the expression levels of genes related to fol-licle growth,proliferation,and apoptosis,including growth differentiation factor 9(Gdf9),zona pellucida glycoprotein 3(Zp3),proliferating cell nuclear antigen(PCNA),Ki-67,B-cell lymphoma 2(BCL2),BCL2-associated X protein(BAX),and caspase-3,with 9 mice in each group.Immunofluorescence staining was used to detect the expression of pro-liferation-and apoptosis-related proteins,including PCNA,Ki-67,5-bromo-2'-deoxyuridine(BrdU),cleaved caspase-3,and forkhead box O3a(FOXO3a),with 15 mice in each group.Western blot was performed to measure the expression of DEAD-box helicase 4(DDX4),phosphorylated mammalian target of rapamycin(p-mTOR),and phosphorylated protein kinase B(p-Akt),with 9 mice in each group.Intraperitoneal injection of CT was administered to 3-dpp mice,and follicle counting was performed with 10 mice in each group.Western blotting was used to detect p-mTOR and p-Akt expression,with 9 mice in each group.Immunofluorescence was employed to assess FOXO3a nuclear export,with 15 mice in each group.For the oral administration of CT in drinking water to 21-dpp mice,immunofluorescence and hematoxylin staining was used to count follicles,with 9 mice in each group.RESULTS:Compared with control group,CT treatment signifi-cantly increased the number of growing follicles in mice.The mRNA and/or protein levels of Gdf9,Zp3,Ki-67,PCNA and DDX4 were markedly elevated.Further studies revealed that CT treatment significantly increased p-Akt levels in the ovaries but had no significant effect on p-mTOR levels.The PI3K/Akt inhibitor LY294002 also reversed the choline the-ophyllinate-induced increase in growing follicles.CONCLUSION:Choline theophyllinate promotes the activation of pri-mordial follicles in mice via the PI3K/Akt signaling pathway in oocytes.
3.Isolation,identification and genetic analysis of a three-lineage NADC30 PRRSV recombinant strain
Bingyan CAI ; Yangyang QIAO ; Tianxin LIU ; Weiyong ZUO ; Yongjuan WANG ; Kai ZHANG ; Hui LU ; Haiming WANG
Chinese Journal of Veterinary Science 2025;45(3):420-426
In June 2018,sows at a pig farm in Jiamusi,Heilongjiang Province,suffered a large num-ber of miscarriages,and subsequently weaned piglets at the farm began to show persistent high fe-ver symptoms at around 35 days of age,with some pigs having a fever of more than 41.5 ℃.In or-der to determine the cause of this outbreak,63 clinical samples from this farm were tested.The re-sults showed that 60 out of 63 samples were positive for the porcine reproductive and respiratory syndrome virus(PRRSV)antigen.Subsequently,PRRSV antigen-positive plasmid was transfected into African green monkey embryonic kidney cells(Marc-145),and after three generations of blind transmission,indirect immunofluorescence assay(IFA)was performed.The results showed that one PRRSV strain,named HLJ38,was successfully isolated.Then the whole genome of HLJ38 strain was sequenced and then analyzed in detail by bioinformatics software.Sequence analysis showed that there were three deletions of 131 amino acids(323-433 aa,483 aa and 504-522 aa)in the derived sequence of Nsp2 gene of HLJ38 strain,which was consistent with the molecular ge-netic marker of NADC30 PRRSV.The phylogenetic tree analysis showed that HLJ38 and NADC30 PRRSV in GenBank belong to lineage 1 subgroup,and the nucleotide homology of HLJ38 and NADC30 PRRSV in GenBank was only 85.2%and 84.6%.Recombinant analysis showed that HLJ38 was a recombinant NADC30-like PRRSV,and the recombinant gene fragments were de-rived from multiple strains,among which the fragment of 1-201 nt was provided by VR2332 strain and fragment of 6 641-8 061 nt derived from the HP-PRRSV strain.In summary,the re-sults showed that the outbreak in this pig farm may be caused by the recombination of PRRSV strains among different lineages,and the recombinant circulating strains not only have certain pathogenicity but also suggest that the existing commercial vaccines provide limited cross-protec-tion against them.Recombination between different lineages increases the genetic diversity of PRRSV and aggravates the difficulty of prevention and control of PRRS in pig farms.Therefore,it is necessary to continuously monitor the epidemic dynamics of PRRSV in pig farms and take effec-tive measures in time to curb the spread of PRRS.
4.Efficacy of reduction robot system combined with minimally invasive, microenvironmental protection, micro-stress shielding fixator in the treatment of Sanders types II and III calcaneal fractures
Xiaohui HAO ; Yongqing WANG ; Zhanmin XU ; Xinan ZHANG ; Zhihui ZHAO ; Jingtao SUN ; Zhiqiang YANG ; Meiyue LIU ; Weiyong WU ; Baoxi HAO ; Juwen CHEN
Chinese Journal of Trauma 2025;41(5):463-470
Objective:To evaluate the efficacy of reduction robot system combined with minimally invasive, microenvironmental protection, micro-stress shielding fixator (short for "3M fixator") for Sanders types II and III calcaneal fractures.Methods:A retrospective case series study was conducted to analyze the clinical data of 26 patients (26 feet) with calcaneal fractures admitted to Fourth Affiliated Central Hospital of Tianjin Medical University from June 2022 to June 2024, including 21 males and 5 females, aged 27-69 years [(46.5±2.5)years]. Among them, 10 patients had fractures in the left foot and 16 in the right. According to the Sanders classification, the fractures were classified as type II in 16 patients and type III in 10. All the patients were treated with the close reduction with reduction robot system combined with 3M fixator in a minimally invasive procedure. The surgical waiting time, operative duration, and fracture healing time were recorded. The length, width and height of the calcaneus, B?hler′s angle and Gissane′s angle were compared before operation and at 1, 3 months after operation. The visual analogue scale (VAS) and American Orthopedic Foot and Ankle Society (AOFAS) ankle-hindfoot score before operation and at 1, 3 months after operation and at the last follow-up were detected. The postoperative complications were recorded.Results:All the patients were followed up for 6-12 months [(9.7±1.1)months]. The surgical waiting time was 1-4 days [(2.0±0.8)days]. The operative duration was 36-66 minutes [(53.2±9.5)minutes]. All the fractures achieved primary union with a healing period of 3-4 months [(3.2±0.4)months]. At 1, 3 months after operation, the length of the calcaneus [(83.3±3.7)mm and (83.6±3.6)mm], width of the calcaneus [(44.3±5.8)mm and (44.3±5.7)mm], height of the calcaneus [(50.1±3.8)mm and (50.3±3.6)mm], B?hler′s angle [(29.8±2.9)° and (29.8±3.0)°], and Gissane angle [(121.1±6.7)° and (123.9±5.9)°] were significantly improved compared with those before operation [(79.3±4.5)mm, (53.6±4.1)mm, (46.2±3.7)mm, (18.9±3.8)°, (109.0±7.5)°, respectively] ( P<0.05), with no significant differences between those indicators at 1, 3 months after operation ( P>0.05). The VAS scores were (3.2±0.6)points, (1.9±0.5)points, and (1.6±0.3)points at 1, 3 months after operation and at the last follow-up, which were lower than (7.1±0.5)points preoperatively and decreased with the prolongation of follow-up time ( P<0.05). The AOFAS scores were (73.5±6.9)points, (90.1±4.3)points, and (92.0±3.6)points, which were higher than (32.0±4.6)points preoperatively and increased with the follow-up time ( P<0.05). One patient had lateral calcaneal pain after operation, and was alleviated after rehabilitation. No complications such as infection or nonunion were found after operation. Conclusion:The reduction robot system combined with 3M fixator for Sanders types II and III calcaneal fractures demonstrates significant clinical advantages, such as reduced surgical waiting time and operative duration, promoted fracture healing, early alleviated pain, enhanced ankle joint functional recovery, and decreased complication occurrence.
5.Development and validation of a deep learning-based low-dose cervical spine X-ray segmentation model
Zhenbo CHEN ; Hongxia ZHANG ; Weiyong YU ; Xinying CONG ; Tian ZHANG ; Yang XIE
Journal of Practical Radiology 2025;41(7):1225-1229
Objective To develop and validate a deep learning-based segmentation model for low-dose cervical spine X-ray,aiming to address the insufficient segmentation accuracy in low-dose protocols while balancing radiation protection and diagnostic accuracy.Methods A total of 1 363 patients cervical spine X-ray images data were collected.A dose-attenuation mathematical simulation sys-tem was constructed to generate 14 122 dynamic low-dose cervical spine images incorporating quantum noise,contrast degradation,and blur artifacts.A neural network model was developed for automated segmentation of low-dose cervical spine X-ray using this dataset.Results Within the dose range of 50%to 7.5%,the average reults of automatic segmentation by the neural network model and manual segmentation for each group were as follows:50%dose group,intersection over union(IoU)=0.98 vs 0.93(P=0.707)and Dice coefficient(Dice)=0.99 vs 0.96(P=0.749);10%dose group,IoU=0.97 vs 0.87(P=0.201)and Dice=0.99 vs 0.93(P=0.219);7.5%dose group,IoU=0.97 vs 0.67(P<0.01)and Dice=0.98 vs 0.80(P<0.01).Conclusion The developed deep learning model achieved robust cervical spine segmentation(IoU>0.96,Dice>0.98)below diagnostic dose thresholds[peak signal-to-noise ratio(PSNR)<38 dB,structural similarity index(SSIM)<0.65].Under ultra-low-dose conditions(PSNR=27.710 dB,SSIM=0.274),it demonstrated a 44.78%IoU improvement and 22.5%Dice improvement over manual segmentation.This model enables minimal radia-tion exposure while preserving diagnostic performance,confirming its theoretical feasibility for low-dose X-ray image analysis and clinical research potential.
6.Atypical forms of microglandular hyperplasia of the cervix: a clinicopathological analysis of 29 cases
Jing ZHAO ; Rongkui LUO ; Lihong ZHANG ; Weiyong GU ; Yinping XIAO ; Xianrong ZHOU ; Xiang TAO ; Yan NING
Chinese Journal of Pathology 2025;54(12):1311-1316
Objective:To investigate the clinicopathological characteristics, immunophenotype and differential diagnosis of atypical forms of microglandular hyperplasia of the cervix (AMGH).Methods:A total of 29 cases of AMGH diagnosed at the Obstetrics and Gynecology Hospital of Fudan University, Shanghai, China from January 2010 to December 2024 were analyzed. Relevant clinical and pathological data of the patients were collected using the electronic medical record system and medical records copied from the outside hospitals. The patients were followed up.Results:Among the 29 cases, 28 were consultation cases, 22 (79%) of the 28 cases were considered as glandular neoplastic lesions by the original institutions. The nature of the lesion was uncertain in 1 case, the diagnosis was suspicious for AMGH in another 1 case, and only 4 cases were clearly diagnosed as AMGH. The median age of the 29 patients was 44 (43, 48) years. Eighteen (62%) of the 29 cases presented as cervical polyp. Twelve of the 16 tested cases were negative for human papillomavirus. The pathological presentation was complex and diverse, including solid, trabecular, cribriform, and papillary patterns, forming pseudo-invasive structures. The glandular epithelium and proliferating reserve cells had diverse morphologies, which presented with abundant eosinophilic cytoplasm or clear cytoplasm. Signet-ring or hobnail cells were also seen. The nuclear atypia was mild, with 0-7 mitotic figures per 10 HPF. Immature squamous metaplasia was noted. The stroma showed edema, myxoid change and hyaline degeneration, accompanied by infiltration of acute and chronic inflammatory cells. Immunohistochemistry demonstrated that p16 was negative in 8/16 of the cases or patchy positive in the other 8/16, Ki-67 positive rate was less than 10% in all 16 cases, p53 was wild phenotype (9/9), and carcinoembryonic antigen was negative in 4/5 cases and focally positive in 1/5 cases, while p63 was positive in 6/9 of the tested cases.Conclusions:AMGH is a benign non-neoplastic lesion of the cervical glands. Half of the cases occur in perimenopausal or postmenopausal women, often presenting as polypoid hyperplasia or localized cervical thickening/elevation with a friable, fragile texture. Microscopically, it may show a pseudoinvasive pattern, making it prone to misdiagnosis as a malignant lesion. Thus, differentiation from cervical adenocarcinoma, clear cell carcinoma and microglandular endometrioid carcinoma is required. Integration of clinical history, immunohistochemistry and molecular testing may aid in the differential diagnosis.
7.Study on the Relationship between the Changes of Four Indexes Related to Plasma Ferroptosis and the Prognosis after TACE in Patients with Hepatocellular Carcinoma
Fei YANG ; Jicheng GAO ; Song LIU ; Huixiao ZUO ; Weiyong GONG ; Zhe ZHANG ; Tao PENG
Journal of Modern Laboratory Medicine 2025;40(5):78-81,87
Objective To analyze the relationship between the expression of ferroptosis markers in the tumor microenvironment(TME)and the prognosis of transcatheter arterial chemoembolization(TACE)for hepatocellular carcinoma(HCC).Methods This prospective observational study included 100 HCC patients who received TACE treatment at Langfang Hospital of Traditional Chinese Medicine from March 2019 to June 2021 as the study subjects.The levels of 8-isoprostaglandin F2α(8-iso-PGF2α),4-hydroxy-2-nonenal(4-HNE),8-hydroxy-2'deoxyguanosine(8-OH-dG)and hepcidin in plasma were evaluated by ELISA kit at baseline(1 day before TACE),1 day after TACE and 4~8 weeks.The changes of ferroptosis related markers during TACE treatment were compared.The difference between the level of 8-iso-PGF2α,4-HNE and the baseline 1 day after TACE treatment was recorded as △8-iso-PGF2α,△4-HNE.Results Compared with the baseline,the levels of 8-iso-PGF2α and 4-HNE increased significantly and the level of hepcidin decreased significantly one day after TACE treatment,and the differences were statistically significant(t=8.03,16.29,2.92,all P<0.05).Compared with 1 day after treatment,the levels of 8-iso-PGF2α,4-HNE decreased and the level of 8-OH-dG increased at 4~8 weeks after TACE treatment,and the differences were statistically significant(t=9.12,17.17,2.63,all P<0.05).Multivariate COX analysis showed that △8-iso-PGF2α,△4-HNE and 8-iso-PGF2α 1 day after TACE treatment were independent factors affecting the overall survival after TACE(Wald χ2=5.205,13.801,6.054,all P<0.05).The survival time of patients with △4-HNE>2.01 μg/ml was significantly longer than that of patients with △4-HNE≤2.01 μg/ml(Log-rank=5.718,P=0.017),and that of patients with△8-iso-PGF2α>1.75ng/ml was sig-nificantly longer than that of patients with△8-iso-PGF2≤1.75ng/ml(Log-rank=4.163,P=0.041).Conclusion The prognosis of HCC patients who are in a state of high ferroptosis(4-HNE and 8-iso-PGF2 increased)at 1 day after TACE treatment is better,which indicate that ferroptosis mediated HCC death induced by TACE treatment.
8.Development and validation of a deep learning-based low-dose cervical spine X-ray segmentation model
Zhenbo CHEN ; Hongxia ZHANG ; Weiyong YU ; Xinying CONG ; Tian ZHANG ; Yang XIE
Journal of Practical Radiology 2025;41(7):1225-1229
Objective To develop and validate a deep learning-based segmentation model for low-dose cervical spine X-ray,aiming to address the insufficient segmentation accuracy in low-dose protocols while balancing radiation protection and diagnostic accuracy.Methods A total of 1 363 patients cervical spine X-ray images data were collected.A dose-attenuation mathematical simulation sys-tem was constructed to generate 14 122 dynamic low-dose cervical spine images incorporating quantum noise,contrast degradation,and blur artifacts.A neural network model was developed for automated segmentation of low-dose cervical spine X-ray using this dataset.Results Within the dose range of 50%to 7.5%,the average reults of automatic segmentation by the neural network model and manual segmentation for each group were as follows:50%dose group,intersection over union(IoU)=0.98 vs 0.93(P=0.707)and Dice coefficient(Dice)=0.99 vs 0.96(P=0.749);10%dose group,IoU=0.97 vs 0.87(P=0.201)and Dice=0.99 vs 0.93(P=0.219);7.5%dose group,IoU=0.97 vs 0.67(P<0.01)and Dice=0.98 vs 0.80(P<0.01).Conclusion The developed deep learning model achieved robust cervical spine segmentation(IoU>0.96,Dice>0.98)below diagnostic dose thresholds[peak signal-to-noise ratio(PSNR)<38 dB,structural similarity index(SSIM)<0.65].Under ultra-low-dose conditions(PSNR=27.710 dB,SSIM=0.274),it demonstrated a 44.78%IoU improvement and 22.5%Dice improvement over manual segmentation.This model enables minimal radia-tion exposure while preserving diagnostic performance,confirming its theoretical feasibility for low-dose X-ray image analysis and clinical research potential.
9.Isolation,identification and genetic analysis of a three-lineage NADC30 PRRSV recombinant strain
Bingyan CAI ; Yangyang QIAO ; Tianxin LIU ; Weiyong ZUO ; Yongjuan WANG ; Kai ZHANG ; Hui LU ; Haiming WANG
Chinese Journal of Veterinary Science 2025;45(3):420-426
In June 2018,sows at a pig farm in Jiamusi,Heilongjiang Province,suffered a large num-ber of miscarriages,and subsequently weaned piglets at the farm began to show persistent high fe-ver symptoms at around 35 days of age,with some pigs having a fever of more than 41.5 ℃.In or-der to determine the cause of this outbreak,63 clinical samples from this farm were tested.The re-sults showed that 60 out of 63 samples were positive for the porcine reproductive and respiratory syndrome virus(PRRSV)antigen.Subsequently,PRRSV antigen-positive plasmid was transfected into African green monkey embryonic kidney cells(Marc-145),and after three generations of blind transmission,indirect immunofluorescence assay(IFA)was performed.The results showed that one PRRSV strain,named HLJ38,was successfully isolated.Then the whole genome of HLJ38 strain was sequenced and then analyzed in detail by bioinformatics software.Sequence analysis showed that there were three deletions of 131 amino acids(323-433 aa,483 aa and 504-522 aa)in the derived sequence of Nsp2 gene of HLJ38 strain,which was consistent with the molecular ge-netic marker of NADC30 PRRSV.The phylogenetic tree analysis showed that HLJ38 and NADC30 PRRSV in GenBank belong to lineage 1 subgroup,and the nucleotide homology of HLJ38 and NADC30 PRRSV in GenBank was only 85.2%and 84.6%.Recombinant analysis showed that HLJ38 was a recombinant NADC30-like PRRSV,and the recombinant gene fragments were de-rived from multiple strains,among which the fragment of 1-201 nt was provided by VR2332 strain and fragment of 6 641-8 061 nt derived from the HP-PRRSV strain.In summary,the re-sults showed that the outbreak in this pig farm may be caused by the recombination of PRRSV strains among different lineages,and the recombinant circulating strains not only have certain pathogenicity but also suggest that the existing commercial vaccines provide limited cross-protec-tion against them.Recombination between different lineages increases the genetic diversity of PRRSV and aggravates the difficulty of prevention and control of PRRS in pig farms.Therefore,it is necessary to continuously monitor the epidemic dynamics of PRRSV in pig farms and take effec-tive measures in time to curb the spread of PRRS.
10.Choline theophyllinate promotes activation of primordial follicles in mice through PI3K/Akt signaling pathway
Tiantian HAO ; Wenbo ZHANG ; Weiyong WANG ; Meijia ZHANG
Chinese Journal of Pathophysiology 2025;41(6):1145-1152
AIM:To investigate the function and mechanism of the smooth muscle relaxant choline theophylli-nate(CT)in activating primordial follicles in mice.METHODS:Experiments were conducted using in vitro culture of 3-day postpartum(dpp)neonatal SPF-grade female mice,intraperitoneal injection in 3-dpp neonatal mice,and oral adminis-tration in 21-dpp adolescent female mice.The mice were divided into control and CT groups.The ovaries were isolated from 3-dpp neonatal mice for the in vitro culture.Hematoxylin staining was used to count the number of primordial and growing follicles,with 10 mice in each group.qPCR was performed to analyze the expression levels of genes related to fol-licle growth,proliferation,and apoptosis,including growth differentiation factor 9(Gdf9),zona pellucida glycoprotein 3(Zp3),proliferating cell nuclear antigen(PCNA),Ki-67,B-cell lymphoma 2(BCL2),BCL2-associated X protein(BAX),and caspase-3,with 9 mice in each group.Immunofluorescence staining was used to detect the expression of pro-liferation-and apoptosis-related proteins,including PCNA,Ki-67,5-bromo-2'-deoxyuridine(BrdU),cleaved caspase-3,and forkhead box O3a(FOXO3a),with 15 mice in each group.Western blot was performed to measure the expression of DEAD-box helicase 4(DDX4),phosphorylated mammalian target of rapamycin(p-mTOR),and phosphorylated protein kinase B(p-Akt),with 9 mice in each group.Intraperitoneal injection of CT was administered to 3-dpp mice,and follicle counting was performed with 10 mice in each group.Western blotting was used to detect p-mTOR and p-Akt expression,with 9 mice in each group.Immunofluorescence was employed to assess FOXO3a nuclear export,with 15 mice in each group.For the oral administration of CT in drinking water to 21-dpp mice,immunofluorescence and hematoxylin staining was used to count follicles,with 9 mice in each group.RESULTS:Compared with control group,CT treatment signifi-cantly increased the number of growing follicles in mice.The mRNA and/or protein levels of Gdf9,Zp3,Ki-67,PCNA and DDX4 were markedly elevated.Further studies revealed that CT treatment significantly increased p-Akt levels in the ovaries but had no significant effect on p-mTOR levels.The PI3K/Akt inhibitor LY294002 also reversed the choline the-ophyllinate-induced increase in growing follicles.CONCLUSION:Choline theophyllinate promotes the activation of pri-mordial follicles in mice via the PI3K/Akt signaling pathway in oocytes.

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