1.The Diversity of Filamentous Morphologies and Magnetic Sensitivity Modulated by Diverse MagR Expression in Bacteria
Ya-Fei CHANG ; Jing ZHANG ; Peng ZHANG ; Xiu-Juan ZHOU ; Meng-Ke WEI ; Tian-Tian CAI ; Pei-Qi HE ; Jun-Feng WANG ; Can XIE
Progress in Biochemistry and Biophysics 2026;53(5):1439-1456
Objective Magnetoreception, the remarkable ability of diverse animals to sense and utilize the geomagnetic field for orientation and navigation, remains a molecularly unresolved mystery in sensory biology. The putative magnetoreceptor (MagR, previously known as IscA1) is a highly conserved iron-sulfur protein implicated in both magnetoreception and iron metabolism; however, the functional diversity among its cross-species homologs remains poorly understood. Cellular morphology is a key genetically determined trait that can be altered through genetic or environmental modifications—a process known as cell morphology engineering. Constructing engineered cells with specific morphological features and magnetic sensitivity to achieve remote, non-invasive magnetic modulation represents a crucial goal in this field with significant application potential. Therefore, this study aims to systematically investigate the effects of MagR heterologous expression on bacterial morphology and magnetic sensing capabilities, screen for MagR-based magnetically sensitive morphology engineering pathways, and reveal the underlying molecular mechanisms. Methods We systematically screened 28 MagR homologous genes from diverse prokaryotic and animal taxa to evaluate their expression and corresponding phenotypic effects in Escherichia coli (E. coli). To compare the differential magnetic responses among bacteria expressing various recombinant MagR proteins, we utilized high-throughput automated bright-field microscopic imaging and scanning electron microscopy (SEM). Furthermore, comprehensive biochemical and biophysical characterizations of iron and iron-sulfur cluster binding were performed using Ferrozine colorimetric assays, electron paramagnetic resonance (EPR) spectroscopy, ultraviolet-visible (UV-Vis) absorption, and circular dichroism (CD) spectroscopy. Additionally, 100 mT static magnetic field (SMF) exposure experiments were conducted to assess magnetically tunable phenotypes, while the intrinsic magnetic properties of purified MagR proteins were directly measured using a superconducting quantum interference device (SQUID) magnetometer. Results Our results demonstrated that the heterologous expression of MagR homologs induced varying degrees of bacterial filamentation. From this comprehensive screen, two distinct morphological patterns were identified: hydra (Hydra vulgaris) MagR (hyMagR) promoted uniform cell elongation and filamentation, exhibiting robust magnetic sensitivity manifested as significantly enhanced filamentation under the 100 mT SMF. In contrast, pigeon (Columba livia) MagR (clMagR) induced only low-frequency, extreme filamentation (sporadically exceeding 80 μm) with a relatively weaker magnetic morphological response. Mechanistically, our data unambiguously proved that these phenotypic differences are primarily driven by distinct iron redox preferences rather than total cellular iron accumulation. Specifically, hyMagR preferentially binds ferrous iron (Fe2+), whereas clMagR favors ferric iron (Fe3+) and forms more stable iron-sulfur clusters. Intriguingly, although SQUID magnetometry showed that purified clMagR exhibited approximately five-fold higher mass magnetic susceptibility than hyMagR, its cellular magnetic response was weaker. We hypothesize that the Fe2+-preferred intracellular environment associated with hyMagR overexpression primes the cell for enhanced generation of reactive oxygen species (ROS) via the Fenton reaction. Exposure to an SMF synergizes with this primed redox state, triggering the bacterial SOS response and upregulating cell division inhibitors to efficiently induce uniform filamentation. Conclusion Our findings identify the Fe2+/Fe3+ redox state as a critical determinant of MagR-mediated morphological remodeling and magnetic responsiveness. This discovery suggests a potential strategy for engineering magnetically responsive cellular systems for synthetic biology applications, and provides a plausible framework, which potentially combines intrinsic protein magnetism with redox-state modulation, for further investigating the evolutionary mechanisms of MagR-mediated magnetoreception.
2.The Diversity of Filamentous Morphologies and Magnetic Sensitivity Modulated by Diverse MagR Expression in Bacteria
Ya-Fei CHANG ; Jing ZHANG ; Peng ZHANG ; Xiu-Juan ZHOU ; Meng-Ke WEI ; Tian-Tian CAI ; Pei-Qi HE ; Jun-Feng WANG ; Can XIE
Progress in Biochemistry and Biophysics 2026;53(5):1439-1456
Objective Magnetoreception, the remarkable ability of diverse animals to sense and utilize the geomagnetic field for orientation and navigation, remains a molecularly unresolved mystery in sensory biology. The putative magnetoreceptor (MagR, previously known as IscA1) is a highly conserved iron-sulfur protein implicated in both magnetoreception and iron metabolism; however, the functional diversity among its cross-species homologs remains poorly understood. Cellular morphology is a key genetically determined trait that can be altered through genetic or environmental modifications—a process known as cell morphology engineering. Constructing engineered cells with specific morphological features and magnetic sensitivity to achieve remote, non-invasive magnetic modulation represents a crucial goal in this field with significant application potential. Therefore, this study aims to systematically investigate the effects of MagR heterologous expression on bacterial morphology and magnetic sensing capabilities, screen for MagR-based magnetically sensitive morphology engineering pathways, and reveal the underlying molecular mechanisms. Methods We systematically screened 28 MagR homologous genes from diverse prokaryotic and animal taxa to evaluate their expression and corresponding phenotypic effects in Escherichia coli (E. coli). To compare the differential magnetic responses among bacteria expressing various recombinant MagR proteins, we utilized high-throughput automated bright-field microscopic imaging and scanning electron microscopy (SEM). Furthermore, comprehensive biochemical and biophysical characterizations of iron and iron-sulfur cluster binding were performed using Ferrozine colorimetric assays, electron paramagnetic resonance (EPR) spectroscopy, ultraviolet-visible (UV-Vis) absorption, and circular dichroism (CD) spectroscopy. Additionally, 100 mT static magnetic field (SMF) exposure experiments were conducted to assess magnetically tunable phenotypes, while the intrinsic magnetic properties of purified MagR proteins were directly measured using a superconducting quantum interference device (SQUID) magnetometer. Results Our results demonstrated that the heterologous expression of MagR homologs induced varying degrees of bacterial filamentation. From this comprehensive screen, two distinct morphological patterns were identified: hydra (Hydra vulgaris) MagR (hyMagR) promoted uniform cell elongation and filamentation, exhibiting robust magnetic sensitivity manifested as significantly enhanced filamentation under the 100 mT SMF. In contrast, pigeon (Columba livia) MagR (clMagR) induced only low-frequency, extreme filamentation (sporadically exceeding 80 μm) with a relatively weaker magnetic morphological response. Mechanistically, our data unambiguously proved that these phenotypic differences are primarily driven by distinct iron redox preferences rather than total cellular iron accumulation. Specifically, hyMagR preferentially binds ferrous iron (Fe2+), whereas clMagR favors ferric iron (Fe3+) and forms more stable iron-sulfur clusters. Intriguingly, although SQUID magnetometry showed that purified clMagR exhibited approximately five-fold higher mass magnetic susceptibility than hyMagR, its cellular magnetic response was weaker. We hypothesize that the Fe2+-preferred intracellular environment associated with hyMagR overexpression primes the cell for enhanced generation of reactive oxygen species (ROS) via the Fenton reaction. Exposure to an SMF synergizes with this primed redox state, triggering the bacterial SOS response and upregulating cell division inhibitors to efficiently induce uniform filamentation. Conclusion Our findings identify the Fe2+/Fe3+ redox state as a critical determinant of MagR-mediated morphological remodeling and magnetic responsiveness. This discovery suggests a potential strategy for engineering magnetically responsive cellular systems for synthetic biology applications, and provides a plausible framework, which potentially combines intrinsic protein magnetism with redox-state modulation, for further investigating the evolutionary mechanisms of MagR-mediated magnetoreception.
3.Reflections on Status Quo and Development Pathways of Traditional Chinese Medicine Technology Transfer in Context of Digital-intelligent Transformation
Jie ZHANG ; Jing XU ; Guangwei ZHENG ; Huayu ZHANG ; Chang LIU ; Xiaoxiao WEN ; Xishui PAN ; Bin WANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(4):235-240
As a distinctive resource of Chinese civilization, traditional Chinese medicine (TCM) technology transfer faces significant opportunities under the background of digital and intelligent transformation, while also being constrained by unique challenges such as the complexity of its theoretical system, lengthy industrial chains, and multidimensional policy restrictions, resulting in a "high-value-high-threshold" paradox. At present, TCM technology transfer is deeply trapped in a "threefold reluctance" dilemma, i.e., unwillingness to transfer, inability to transfer, and lack of capacity to transfer. Specifically, the disconnection between scientific research evaluation systems and market demand leads to low conversion rates of research achievements, unclear ownership and compliance risks suppress innovation incentives, and the absence of professional services intensifies supply-demand mismatches. This article systematically analyzes the specific characteristics of TCM technology transfer and proposes a breakthrough pathway centered on full-chain digital and intelligent transformation. By integrating technologies such as intelligent sorting systems, blockchain-based traceability, and AI diagnostic models, the TCM ecosystem spanning "cultivation-production-service" can be reconstructed. In terms of standardization, promoting the progression from "experience-based data conversion" to "data standardization" and further to "intelligent standardization" is advocated to resolve quality control challenges. For example, a "three-no-one-full" certification system can strengthen quality trust. Policy coordination should focus on optimizing mechanisms for the transformation of scientific and technological achievements, while exploring intellectual property securitization and risk-sharing models to stimulate research momentum. In terms of internationalization, reliance on the Belt and Road Initiative platform to promote the export of geo-authentic medicinal material brands and standards is recommended to build a dual-driven model of "technology plus culture". Looking ahead, through the construction of national-level databases, the cultivation of interdisciplinary talent, and the mutual recognition of international standards, a new paradigm of "scientific intelligent manufacturing" can be formed, providing systematic solutions for the modernization of TCM and global health governance.
4.Effect of fatty liver on cardiac structure and function: a cross-sectional study based on health examination
Peiwen CHEN ; Xingxing REN ; Hongmei YAN ; Xinxia CHANG ; Jing ZHANG ; Hailuan ZENG ; Jingjing JIANG
Chinese Journal of Clinical Medicine 2026;33(3):434-444
Objective To investigate the cross-sectional associations between different fatty liver classifications and cardiac structure and function in people undergo health examination. Methods A total of 6 545 adults who underwent health examinations at the Health Management Center of Zhongshan Hospital, Fudan University between January 1 and December 31, 2017, were retrospectively included. Demographic characteristics and laboratory data were collected. The hepatic steatosis was graded by ultrasonography. And patients with fatty liver were further stratified according to alanine aminotransferase (ALT) or aspartate aminotransferase (AST) levels, as well as the degree of liver fibrosis. Cardiac morphology and function were assessed by transthoracic echocardiography, and left ventricular geometric patterns were classified accordingly. Multivariate logistic regression analysis was performed to evaluate the associations between fatty liver classifications and cardiac abnormalities. Results There were 2 795 patients (42.7%) with fatty liver, of whom 832 (29.8%) had significant cardiac structural alterations and 1 500 (53.7%) had diastolic dysfunction. Severe fatty liver was risk factor for concentric remodeling and increased relative wall thickness (RWT), with odds ratios (ORs) of 1.27 (P=0.012) and 1.27 (P=0.009), respectively. Fatty liver accompanied by elevated ALT or AST was risk factor for concentric remodeling (OR=1.45,1.56; P=0.001, 0.001), increased RWT (OR=1.48,1.57; P<0.001, <0.001), and diastolic dysfunction (OR=1.27, 1.32; P=0.035, 0.040), respectively. Fatty liver with liver fibrosis was risk factor for concentric remodeling (OR=1.83, P=0.046) and diastolic dysfunction (OR=2.64, P=0.034). Conclusions Advanced fatty liver, including severe hepatic steatosis, accompanied by elevated liver enzymes or liver fibrosis, could increase risks of cardiac remodeling and diastolic dysfunction, while systolic function is preserved. For patients with fatty liver, it is recommended to undergo regular ultrasonography examination.
5.Correlation between walking exercise guided by walking test and long-term prognosis of acute coronary syndrome in the elderly
Yi MA ; Jing HAN ; Wenhong CHANG ; Shumei ZHENG ; Jianxiu DONG ; Hongxin ZHANG ; Lili HU ; Jianhui WANG ; Xuebin GENG
Chinese Journal of Geriatric Heart Brain and Vessel Diseases 2025;27(6):693-697
Objective To explore the association between walking exercise guided by 6 minute walking test(6MWT)and the incidences of 3-year major adverse cardiovascular event(MACE)in elderly patients with acute coronary syndrome(ACS)after percutaneous coronary intervention(PCI).Methods A total of 628 elderly ACS patients who undergoing PCI and obtaining success-ful coronary revascularization in our department from November 2018 to April 2019 were enrolled,and divided into 6MWT group(n=147)and control group(n=481)based on participa-ting in walking exercise guided by 6MWT or not.All of them were followed up for 3 years.The incidences of MACE[including coronary target vascular restenosis,acute myocardial infarction,heart failure,ischemic or hemorrhagic stroke]and all-cause death were observed.Univariate and multivariate Cox proportional analyses and Kaplan-Meier survival curve analysis were employed for data statistical analyses.Results At the end of follow-up,the incidences of target vascular restenosis(6.9%vs 2.0%,P=0.028),heart failure(3.7%vs 0%,P=0.036),stroke(3.7%vs 0%,P=0.036),and total MACE incidence(15.0%vs 4.1%,P=0.000)were statistically higher in the control group than the 6MWT group.Kaplan-Meier survival curve analysis showed that the cumulative incidence of MACE was significantly lower in the 6MWT group than the control group(Plog rank=0.001).Multivariate Cox regression analysis showed that not participating in walking exercise guided by 6MWT was an independent risk factor for occurrence of 3-year MACE(HR=3.102,95%CI:1.327-7.250,P=0.009).Conclusion Walking exercise guided by 6MWT reduces the incidence of 3-year MACE and improves the long-term prognosis of elderly ACS patients after PCI.
6.Study on Mechanism of Xuefu Zhuyu Decoction in Interfering Oxidative Stress Injury in Rats with Heart Blood Stasis Syndrome of Coronary Heart Disease Based on Keap1/Nrf2 Signaling Pathway
Huifang KUANG ; Jing LI ; Peng TIAN ; Chang SU ; Yi LIU ; Mingyun WANG ; Qiuyan ZHANG
Chinese Journal of Information on Traditional Chinese Medicine 2025;32(7):104-111
Objective To investigate the effects and mechanism of Xuefu Zhuyu Decoction in oxidative stress in coronary heart disease model rats with heart blood stasis syndrome based on Keap1/Nrf2 signaling pathway.Methods The rats were divided into normal group,sham-operation group,model group,Xuefu Zhuyu Decoction group and trimetazidine group.The rat model of coronary heart disease with heart blood stasis syndrome was established by ligation of the left anterior descending branch of the coronary artery.Xuefu Zhuyu Decoction group and trimetazidine group were administrated with the corresponding drugs at the dosages of 14.04 g/kg and 5.4 mg/kg,respectively,and normal group,sham-operation group and model group were administrated with the same volume of normal saline for 14 days.The general state of rats was observed,body mass was recorded and electrocardiogram was collected.Echocardiography was used to examine cardiac functions(LVEF,LVFS,LVIDd,LVIDs);the morphology of myocardial tissue was observed by HE staining,serum malondialdehyde(MDA),superoxide dismutase(SOD),glutathione peroxidase(GSH-Px)and total antioxidant capacity(T-AOC)were detected by ELISA,the positive expressions of Keap1,Nrf2,HO-1 and NQO1 in myocardial tissue were detected by immunohistochemistry.Results Compared with the normal group and sham-operation group,the rats in the model group showed signs of mental fatigue,reduced activity,dull fur,purple claws,and a significant decrease in body mass(P<0.01);the ST segment in lead Ⅱ of the electrocardiogram was significantly elevated,LVEF and LVFS were significantly reduced,and LVIDd and LVIDs significantly increased(P<0.01),with severe degeneration and necrosis of myocardial cells,disappearance of striated structures,disordered arrangement of myocardial fibers,infiltration of inflammatory cells;the serum MDA content significantly increased,while the activities of SOD,GSH-Px and T-AOC significantly decreased(P<0.01);the positive expressions of Keap1 and Nrf2 in myocardial tissue significantly increased,while the positive expression of HO-1 and NQO1 significantly decreased(P<0.01).Compared with the model group,the rats in Xuefu Zhuyu Decoction group and trimetazidine group showed improvement in their mental state,increased activity,shiny fur,rosy nails,and significantly increased body mass(P<0.01);the ST segment of the electrocardiogram decreased to varying degrees,with significant increases in LVEF and LVFS,and significant decreases in LVIDd and LVIDs(P<0.01);a large number of myocardial cells survived,the arrangement of myocardial fibers was relatively regular,and the infiltration of inflammatory cells was significantly reduced;the serum MDA content was significantly reduced,while the activities of SOD,GSH-Px and T-AOC significantly increased(P<0.01);the positive expression of Keap1 in myocardial tissue significantly decreased,while the positive expressions of Nrf2,HO-1 and NQO1 significantly increased(P<0.01).Conclusion Xuefu Zhuyu Decoction may inhibit oxidative stress by activating Keap1/Nrf2 signaling pathway to improve the pathological morphology and structural damage of myocardial tissue and promote the recovery of cardiac functions in rats with heart blood stasis syndrome of coronary heart disease.
7.Correlation between walking exercise guided by walking test and long-term prognosis of acute coronary syndrome in the elderly
Yi MA ; Jing HAN ; Wenhong CHANG ; Shumei ZHENG ; Jianxiu DONG ; Hongxin ZHANG ; Lili HU ; Jianhui WANG ; Xuebin GENG
Chinese Journal of Geriatric Heart Brain and Vessel Diseases 2025;27(6):693-697
Objective To explore the association between walking exercise guided by 6 minute walking test(6MWT)and the incidences of 3-year major adverse cardiovascular event(MACE)in elderly patients with acute coronary syndrome(ACS)after percutaneous coronary intervention(PCI).Methods A total of 628 elderly ACS patients who undergoing PCI and obtaining success-ful coronary revascularization in our department from November 2018 to April 2019 were enrolled,and divided into 6MWT group(n=147)and control group(n=481)based on participa-ting in walking exercise guided by 6MWT or not.All of them were followed up for 3 years.The incidences of MACE[including coronary target vascular restenosis,acute myocardial infarction,heart failure,ischemic or hemorrhagic stroke]and all-cause death were observed.Univariate and multivariate Cox proportional analyses and Kaplan-Meier survival curve analysis were employed for data statistical analyses.Results At the end of follow-up,the incidences of target vascular restenosis(6.9%vs 2.0%,P=0.028),heart failure(3.7%vs 0%,P=0.036),stroke(3.7%vs 0%,P=0.036),and total MACE incidence(15.0%vs 4.1%,P=0.000)were statistically higher in the control group than the 6MWT group.Kaplan-Meier survival curve analysis showed that the cumulative incidence of MACE was significantly lower in the 6MWT group than the control group(Plog rank=0.001).Multivariate Cox regression analysis showed that not participating in walking exercise guided by 6MWT was an independent risk factor for occurrence of 3-year MACE(HR=3.102,95%CI:1.327-7.250,P=0.009).Conclusion Walking exercise guided by 6MWT reduces the incidence of 3-year MACE and improves the long-term prognosis of elderly ACS patients after PCI.
8.Study on Mechanism of Xuefu Zhuyu Decoction in Interfering Oxidative Stress Injury in Rats with Heart Blood Stasis Syndrome of Coronary Heart Disease Based on Keap1/Nrf2 Signaling Pathway
Huifang KUANG ; Jing LI ; Peng TIAN ; Chang SU ; Yi LIU ; Mingyun WANG ; Qiuyan ZHANG
Chinese Journal of Information on Traditional Chinese Medicine 2025;32(7):104-111
Objective To investigate the effects and mechanism of Xuefu Zhuyu Decoction in oxidative stress in coronary heart disease model rats with heart blood stasis syndrome based on Keap1/Nrf2 signaling pathway.Methods The rats were divided into normal group,sham-operation group,model group,Xuefu Zhuyu Decoction group and trimetazidine group.The rat model of coronary heart disease with heart blood stasis syndrome was established by ligation of the left anterior descending branch of the coronary artery.Xuefu Zhuyu Decoction group and trimetazidine group were administrated with the corresponding drugs at the dosages of 14.04 g/kg and 5.4 mg/kg,respectively,and normal group,sham-operation group and model group were administrated with the same volume of normal saline for 14 days.The general state of rats was observed,body mass was recorded and electrocardiogram was collected.Echocardiography was used to examine cardiac functions(LVEF,LVFS,LVIDd,LVIDs);the morphology of myocardial tissue was observed by HE staining,serum malondialdehyde(MDA),superoxide dismutase(SOD),glutathione peroxidase(GSH-Px)and total antioxidant capacity(T-AOC)were detected by ELISA,the positive expressions of Keap1,Nrf2,HO-1 and NQO1 in myocardial tissue were detected by immunohistochemistry.Results Compared with the normal group and sham-operation group,the rats in the model group showed signs of mental fatigue,reduced activity,dull fur,purple claws,and a significant decrease in body mass(P<0.01);the ST segment in lead Ⅱ of the electrocardiogram was significantly elevated,LVEF and LVFS were significantly reduced,and LVIDd and LVIDs significantly increased(P<0.01),with severe degeneration and necrosis of myocardial cells,disappearance of striated structures,disordered arrangement of myocardial fibers,infiltration of inflammatory cells;the serum MDA content significantly increased,while the activities of SOD,GSH-Px and T-AOC significantly decreased(P<0.01);the positive expressions of Keap1 and Nrf2 in myocardial tissue significantly increased,while the positive expression of HO-1 and NQO1 significantly decreased(P<0.01).Compared with the model group,the rats in Xuefu Zhuyu Decoction group and trimetazidine group showed improvement in their mental state,increased activity,shiny fur,rosy nails,and significantly increased body mass(P<0.01);the ST segment of the electrocardiogram decreased to varying degrees,with significant increases in LVEF and LVFS,and significant decreases in LVIDd and LVIDs(P<0.01);a large number of myocardial cells survived,the arrangement of myocardial fibers was relatively regular,and the infiltration of inflammatory cells was significantly reduced;the serum MDA content was significantly reduced,while the activities of SOD,GSH-Px and T-AOC significantly increased(P<0.01);the positive expression of Keap1 in myocardial tissue significantly decreased,while the positive expressions of Nrf2,HO-1 and NQO1 significantly increased(P<0.01).Conclusion Xuefu Zhuyu Decoction may inhibit oxidative stress by activating Keap1/Nrf2 signaling pathway to improve the pathological morphology and structural damage of myocardial tissue and promote the recovery of cardiac functions in rats with heart blood stasis syndrome of coronary heart disease.
9.Research Progress of 223-Ra in the Treatment of Bone Metastases from Desmoplasia-resistant Prostate Cancer
Chang LU ; Ran ZHANG ; Li ZHANG ; Jiaxin DING ; Yue SUN ; Zhuoling RAN ; Yuxuan ZHENG ; Lin YU ; Xu GAO ; Jing XIE ; Huan ZHOU ; Jian GONG
Herald of Medicine 2025;44(3):446-451
Prostate cancer is one of the most common male urological malignancies,in which bone metastasis of desmo-plasia-resistant prostate cancer is an important stage in the progression of the disease,which seriously affects the quality of life and survival of patients.With the development of nuclide therapy technology in recent years,223-Ra,as a new type of alpha-targeted therapy,has shown good efficacy in the treatment of desmoplasia-resistant prostate cancer bone metastasis.The purpose of this pa-per is to review the characteristics,mechanism of action,treatment,and the main research results of its treatment of desmoplasia-resistant prostate cancer bone metastasis,and provide a comprehensive review of the clinical application of 223-Ra in the treatment of desmoplasia-resistant prostate cancer bone metastasis for the clinical application of 223-Ra in prostate cancer bone metastasis.
10.Study on the repair of ovarian tissue injury in mice in simulated plateau environment by human umbilical cord mesenchymal stem cell transplantation combined with high-pressure medical treatment
Jing DU ; Ruiya CHANG ; Zongcui ZHANG ; Ling WANG
Chinese Journal of Reproduction and Contraception 2025;45(2):183-195
Objective:To investigate the reparative effects and underlying mechanisms of human umbilical cord mesenchymal stem cells (hUCMSCs) in conjunction with hyperbaric oxygen (HBO) on ovarian dysfunction in mice exposed to simulated high-altitude conditions.Methods:A total of 64 six-week-old female C57BL/6 mice were randomly allocated according to random number table, assigned to control group, model group, hUCMSCs group and hUCMSCs+HBO group, with 16 mice in each group. Mice in model group, hUCMSCs group and hUCMSCs+HBO group were placed in a hypobaric oxygen chamber to simulate a high-altitude environment at an elevation of 6 500 m for 15 d, thereby establishing a model of ovarian dysfunction. Beginning on the first day following model establishment, the control and model groups received intravenous injections of 0.2 mL saline via the tail vein. In contrast, the hUCMSCs group and the hUCMSCs+HBO group received 0.2 mL hUCMSCs (1×10 6 cells) per mouse through tail vein injection, administered once a week for three weeks, with continuous intervention lasting 15 d. Furthermore, the hUCMSCs+HBO group were subjected to daily hyperbaric oxygen treatment. The recorded variables included general health status, body weight, estrous cycle changes, serum hormone levels, bilateral ovarian wet weight, ovarian index, follicular development assessment, pathological alterations of ovarian tissue, ultrastructural changes, and the expression levels of transforming growth factor beta 1 (TGF-β1) and Smad family member 3 (Smad3) detected by Western blotting in ovarian tissue. Additionally, litter size and offspring body weight were measured across all groups to evaluate the reproductive capacity of the mice. Results:1) Compared with the hUCMSCs group, the hUCMSCs+HBO group exhibited no statistically significant differences in estrous cycle, body weight, bilateral ovarian wet weight, or ovarian index (all P>0.05). Furthermore, serum levels of anti-Müllerian hormone (AMH), estradiol, progesterone, follicle-stimulating hormone (FSH), and luteinizing hormone (LH), as well as counts of various stages of follicles in hUCMSCs+HBO group, did not demonstrate statistically significant differences compared with hUCMSCs group (all P>0.05). Notably, in the hUCMSCs+HBO group, the integrity of the nuclear membrane in granulosa cells of the ovarian tissue was preserved, with only mild swelling observed in individual mitochondria, while no expansion of the rough endoplasmic reticulum or swelling of the Golgi apparatus were noted. Additionally, expression levels of TGF-β1 and Smad3 proteins in the hUCMSCs+HBO group were significantly elevated compared with the hUCMSCs group ( P=0.010, P<0.001). The conditions of the offspring and litter size showed no statistically significant differences between the two groups (all P>0.05). 2)Compared with control group, the hUCMSCs+HBO group had slightly lower values for body weight, bilateral ovarian wet weight, ovarian index, serum levels of AMH, estradiol, and progesterone, while had slightly higher serum levels of FSH and LH without statistically significant differences (all P>0.05). Importantly, the ultrastructural characteristics of granulosa cells in the ovarian tissue of the hUCMSCs+HBO group closely resembled those of control group, displaying intact structures of the nuclear membrane, mitochondria, rough endoplasmic reticulum, and Golgi apparatus. There was no significant difference in TGF-β1 protein level between hUCMSCs+HBO group and control group ( P=0.253), but Smad3 protein level in hUCMSCs+HBO group was higher than that in control group ( P<0.001). No statistically significant differences were detected in offspring body weight, litter size, or behavioral tendencies (all P>0.05). Conclusion:Both hUCMSCs and the combination of hUCMSCs with HBO intervention demonstrated a safe and effective promotion of functional repair in damaged ovarian tissue under hypoxic conditions. Notably, the combination treatment of hUCMSCs with HBO exhibited a synergistic effect compared with hUCMSCs alone. The potential mechanism underlying this enhanced functional repair may involve the upregulation of the TGF-β1/Smad3 signaling pathway, which could ultimately improve fertility outcomes in mice subjected to hypoxic environments.

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