1.Effect of Feiyanning Granules on Inducing Ferroptosis in Lung Cancer Cells and Its Regulatory Function onNrf2/SLC7A11/GPX4 Signaling Pathway
Xin LIU ; Wenjie WANG ; Zhenye XU ; Zhan ZHENG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(2):100-107
ObjectiveThis study aims to explore the effect of Feiyanning granules on ferroptosis in lung cancer cells and its regulatory function within the nuclear transcription factor E2-related factor 2 (Nrf2)/mouse solute carrier family 7 member 11 (SLC7A11)/glutathione peroxidase 4 (GPX4) signaling pathway. MethodsThe cell counting kit-8 (CCK-8) method was used to detect the effect of Feiyanning granule on the proliferation of A549 lung cancer cells. A549 lung cancer cells were categorized into a blank group, a ferroptosis inhibitor-1 (Fer-1) group (10 μmol·L-1), a Feiyanning granules (600 mg·L-1) group, and a Feiyanning granules + Fer-1 group. After 48 hours of intervention, the activity and morphology of the cells were observed. The CCK-8 method was employed to measure cell viability. Biochemical assays were carried out to measure the levels of reactive oxygen species (ROS), malondialdehyde (MDA), glutathione (GSH), and ferrous ions (Fe²⁺) in A549 cells. Western blot was utilized to evaluate the expression levels of Kelch-like ECH-associated protein 1 (Keap1), Nrf2, SLC7A11, and GPX4 proteins. A549 lung cancer cells were categorized into a blank group and a Feiyanning Granule group (600 mg·L-1), and mitochondrial morphology was examined via transmission electron microscopy (TEM). ResultsAfter the intervention of Feiyaning granules, the proliferation of A549 cells was significantly inhibited in a concentration-dependent manner compared with that in the blank group (P<0.01). Compared with the blank group, the Feiyanning granules group exerted an significantly inhibitory effect on the viability of lung cancer cells (P<0.01). Compared with that in the Feiyanning granules group, the cell viability in the Feiyanning granules +Fer-1 group was obviously restored (P<0.05). Compared with the blank group, the Feiyanning Granule group showed a significant increase in the levels of ROS, MDA, and Fe²⁺ (P<0.01), a significant decrease in the GSH level (P<0.01), and facilitated ferroptosis. Compared with the blank group, the Feiyanning granules group showed significantly decreased expression of Nrf2, SLC7A11, and GPX4 proteins and enhanced expression of Keap1 (P<0.01). Compared with those in the Feiyanning Granule group, the protein levels of Nrf2, SLC7A11, and GPX4 increased significantly (P<0.01), and the expression of Keap1 decreased significantly in the Feiyanning granules + Fer-1 group (P<0.01). Compared with the blank group, the Feiyaning granules group exhibited reduced mitochondrial size and increased matrix electron density. ConclusionFeiyanning granules can induce ferroptosis in lung cancer cells, and its underlying mechanism might be associated with the inhibition of the Nrf2/SLC7A11/GPX4 signaling pathway.
2.Comparison of Wild and Cultivated Gardeniae Fructus Based on Traditional Quality Evaluation
Yuanjun SHANG ; Bo GENG ; Xin CHEN ; Qi WANG ; Guohua ZHENG ; Chun LI ; Zhilai ZHAN ; Junjie HU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(5):225-234
ObjectiveBased on traditional quality evaluation of Gardeniae Fructus(GF) recorded in historical materia medica, this study systematically compared the quality differences between wild and cultivated GF from morphological characteristics, microscopic features, and contents of primary and secondary metabolites. MethodsVernier calipers and analytical balances were used to measure the length, diameter and individual fruit weight of wild and cultivated GF, and the aspect ratio was calculated. A colorimeter was used to determine the chromaticity value of wild and cultivated GF, and the paraffin sections of them were prepared by safranin-fast green staining and examined under an optical microscope to observe their microstructure. Subsequently, the contents of water-soluble and alcohol-soluble extracts of wild and cultivated GF were detected by hot immersion method under the general rule 2201 in volume Ⅳ of the 2020 edition of the Pharmacopoeia of the People's Republic of China, the starch content was measured by anthrone colorimetric method, the content of total polysaccharides was determined by phenol-sulfuric acid colorimetric method, the sucrose content was determined by high performance liquid chromatography coupled with evaporative light scattering detection(HPLC-ELSD), and the contents of representative components in them were measured by ultra-performance liquid chromatography(UPLC). Finally, correlation analysis was conducted between quality traits and phenotypic traits, combined with multivariate statistical analysis methods such as principal component analysis(PCA) and orthogonal partial least squares-discriminant analysis(OPLS-DA), key differential components between wild and cultivated GF were screened. ResultsIn terms of traits, the wild GF fruits were smaller, exhibiting reddish yellow or brownish red hues with significant variation between batches. While the cultivated GF fruits are larger, displaying deeper orange-red or brownish red. The diameter and individual fruit weight of cultivated GF were significantly greater than those of wild GF, while the blue-yellow value(b*) of wild GF was significantly higher than that of cultivated GF. In the microstructure, the mesocarp of wild GF contained numerous scattered calcium oxalate cluster crystals, while the endocarp contained stone cell class round, polygonal or tangential prolongation, undeveloped seeds were visible within the fruit. In contrast, the mesocarp of cultivated GF contained few calcium oxalate cluster crystals, or some batches exhibited extremely numerous cluster crystals. The stone cells in the endocarp were predominantly round-like, with the innermost layer arranged in a grid pattern. Seeds were basically mature, and only a few immature seeds existed in some batches. Regarding primary metabolite content, wild GF exhibited significantly higher total polysaccharide level than cultivated GF(P<0.01). In category-specific component content, wild GF exhibited significantly higher levels of total flavonoids and total polyphenols compared to cultivated GF(P<0.01). Analysis of 12 secondary metabolites revealed that wild GF exhibited significantly higher levels of Shanzhiside, deacetyl asperulosidic acid methyl ester, gardenoside and chlorogenic acid compared to cultivated GF(P<0.01). Conversely, the contents of genipin 1-gentiobioside, geniposide and genipin were significantly lower in wild GF(P<0.01). ConclusionThere are significant differences between wild and cultivated GF in terms of traits, microstructure, and contents of primary and secondary metabolites. At present, the quality evaluation system of cultivated GF remains incomplete, and this study provides a reference for guiding the production of high-quality GF medicinal materials.
3.Construction of Perimenopausal Depression Animal Models and Mechanism of Action of Traditional Chinese Medicine: A Review
Xiaoting LI ; Shouzhu XU ; Jun KE ; Zhan ZHANG ; Bao XIN
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(4):258-267
Perimenopausal depression (PMD) is an affective disorder that occurs in women during the transition from sexual maturity to old age. It can induce various complications, such as insomnia and cognitive decline. The etiology of PMD is complex. Although multiple hypotheses have been proposed, there is still no unified theory that fully explains its pathogenesis. Research into its mechanisms relies heavily on animal experiments, and establishing reliable animal models is crucial for experimental studies. Appropriate animal models can better simulate human pathophysiological states, rapidly evaluate the efficacy and safety of drugs and intervention methods, grasp the essence of the disease, and uncover its intrinsic connections, thereby exploring more advanced intervention strategies. However, there is a lack of systematic review and summarization of literature related to model construction. Additionally, traditional Chinese medicine (TCM), adhering to the principles of ''syndrome differentiation and treatment'' and ''holistic concept'', has shown significant efficacy in treating PMD. In recent years, research exploring and analyzing its therapeutic mechanisms has been increasing. Therefore, to gain a clearer and more comprehensive understanding of PMD animal modeling methods and the mechanisms of TCM, this paper reviewed Chinese and English literature on PMD animal models and mechanisms of TCM in PMD treatment. It summarized the construction methods of single-factor and multi-factor PMD models, and discussed the advantages and disadvantages of each modeling approach. Furthermore, it delved into the mechanisms of TCM intervention in PMD, revealing that TCM formulas primarily exert their effects by regulating the hypothalamic-pituitary-adrenal axis, hypothalamic-pituitary-ovarian axis, gut-brain axis, cell signaling pathways, neural circuits, hormone levels, and neurotransmitter levels. This review aims to provide a reference for future research in this field. In summary, by summarizing the progress in the methods for PMD animal model construction and the mechanisms of TCM, the paper seeks to offer new insights into the mechanistic research of TCM intervention in PMD.
4.Evaluation of Pulmonary Air-Blood Barrier Damage in Ulcerative Colitis Inflammatory Cancer Transformation Model Mice:Based on the "Lung-Intestine Correlation" Theory
Huiyan XU ; Haimei ZHANG ; Xinyu ZHAN ; Fanwu WU ; Yongsen JIA ; Chenxi WU ; Lingyu KONG ; Xin YAN
Journal of Traditional Chinese Medicine 2026;67(7):776-783
ObjectiveTo dynamically observe and evaluate the damage to the pulmonary air-blood barrier in mice during the inflammatory cancer transformation process of ulcerative colitis (UC) based on the "lung-intestine correlation" theory. MethodsSixty-five C57BL/6 mice were divided into a normal group (n=25) and a model group (n=40) using a random number table. Azoxymethane/dextran sodium sulfate (DSS) method was used to establish a mouse model of UC inflammation cancer transformation in the modeling group. According to the tissue collection time points at 5, 8, 11, 13, and 15 weeks, the normal group mice were randomly divided into the normal 5w, 8w, 11w, 13w, and 15w groups. The model group mice, 10 mice of which died after the first cycle of DSS administration, were randomly divided into model 5w, 8w, 11w, 13w, and 15w groups. During the experiment, the general condition of the mice was observed daily, and their body weight was measured weekly. At the corresponding tissue collection time points, the colon length of each group was measured. Histopathology of mouse lung and colon tissues was examined using HE staining. Immunofluorescence was used to detect changes in the positive expression of tight junction protein (ZO-1), vascular endothelial cadherin (VE-cadherin), and cytoskeletal protein (F-actin) in lung and colon tissues. RT-PCR was used to detect the mRNA expression of apoptosis regulatory proteins B-cell lymphoma-2 (Bcl-2), BCL2-associated X protein (Bax), and Cysteine aspartic acid protease-3 (Caspase-3) in lung tissues. Western Blot was employed to measure protein levels of ZO-1, VE-cadherin, and F-actin in lung tissues. ResultsCompared to the normal group at the same time point, the mice in the model group at each time point generally had poorer conditions, with weight loss and shortened colon length (P<0.05 or P<0.01). In the model 5w group, there was significant inflammatory cell infiltration in the colon tissue; in the model 8w group, there was mild atypical hyperplasia; in the model 11w group, the crypt structure was disordered, and moderate to severe atypical hyperplasia occurred; in the model 13w and 15w groups, tumors appeared. Pulmonary interstitial lesions, inflammation, vasculitis, and fibrosis were observed at all stages of UC inflammation cancer transformation. The protein levels of ZO-1, VE-cadherin, and F-actin, as well as Bcl-2 mRNA expression in lung tissue decreased during the acute inflammatory recovery period, atypical hyperplasia period, and canceration period, while the expressions of Bax and Caspase-3 mRNA increased; the expressions of ZO-1, VE-cadherin, and F-actin proteins in colon tissue decreased during the acute inflammatory recovery period, atypical hyperplasia period, and canceration period (P<0.01 or P<0.05). Compared to the model 5w group, the ZO-1 and F-actin protein levels and Bcl-2 mRNA expression in lung tissue in the other model groups increased in the atypical hyperplasia period and canceration period, while the expressions of Bax and Caspase-3 mRNA decreased; the expression of ZO-1 protein in colon tissue increased in the canceration period, and the expression of VE-cadherin protein decreased in the atypical hyperplasia period (P<0.01 or P<0.05). ConclusionIn the process of "inflammatory response-atypical hyperplasia-carcinogenesis" in UC inflammatory cancer transformation mice, there were damage to air-blood barrier.
5.Clinical analysis of 6 cases of diffuse panbronchiolitis in children.
Li-Xin DENG ; De-Hui CHEN ; Yu-Neng LIN ; Shang-Zhi WU ; Jia-Xing XU ; Zhan-Hang HUANG ; Ying-Ying GU ; Jun-Xiang FENG
Chinese Journal of Contemporary Pediatrics 2025;27(3):334-339
OBJECTIVES:
To analyze the clinical characteristics of diffuse panbronchiolitis (DPB) in children and to enhance the clinical diagnosis and treatment of this disease.
METHODS:
A retrospective analysis was conducted on the clinical data of 6 children diagnosed with DPB who were hospitalized at The First Affiliated Hospital of Guangzhou Medical University from January 2011 to December 2019.
RESULTS:
Among the 6 patients, there were 2 males and 4 females; the age at diagnosis ranged from 7 to 12 years. All patients presented with cough, sputum production, and exertional dyspnea, and all had a history of sinusitis. Two cases showed positive serum cold agglutinin tests, and 5 cases exhibited pathological changes consistent with chronic bronchiolitis. High-resolution chest CT in all patients revealed centrilobular nodules diffusely distributed throughout both lungs with a tree-in-bud appearance. Five patients received low-dose azithromycin maintenance therapy, but 3 showed inadequate treatment response. After empirical anti-tuberculosis treatment, non-tuberculous Mycobacteria were found in the bronchoalveolar lavage fluid. Follow-up over 2 years showed 1 case cured, 3 cases significantly improved, and 2 cases partially improved.
CONCLUSIONS
The clinical presentation of DPB is non-specific and can easily lead to misdiagnosis. In cases where DPB is clinically diagnosed but does not show improvement with low-dose azithromycin treatment, special infections should be considered.
Humans
;
Male
;
Female
;
Bronchiolitis/drug therapy*
;
Retrospective Studies
;
Child
;
Haemophilus Infections/diagnosis*
6.Clinical Applications of Circulating Tumor DNA in Response Evaluation and Relapse Monitoring of Primary Mediastinal Large B-Cell Lymphoma.
Lu PAN ; Xin-Miao JIANG ; Yan TENG ; Ning WANG ; Ling HUANG ; Han-Guo GUO ; Si-Chu LIU ; Xiao-Juan WEI ; Fei-Li CHEN ; Zhan-Li LIANG ; Wen-Yu LI
Journal of Experimental Hematology 2025;33(2):407-415
OBJECTIVE:
To explore the clinical significance of circulating tumor DNA (ctDNA) in response evaluation and relapse monitoring for patients with primary mediastinal large B-cell lymphoma (PMBCL).
METHODS:
The clinical characteristics, efficacy and survival of 38 PMBCL patients in our hospital from January 2010 to April 2020 were retrospectively analyzed. The ctDNA monitoring was conducted by targeted next-generation sequencing (NGS).
RESULTS:
Among the 38 patients, 26 cases were female, and 32 cases were diagnosed with Ann Arbor stage I-II. The 5-year overall survival (OS) rate and progression-free survival (PFS) rate were 74.7% and 61.7%, respectively. Males and those with high aaIPI scores (3 points) had a relatively poor prognosis. The NGS results of 23 patients showed that STAT6 (65.2%), SOCS1 (56.5%), and TNFAIP3 (56.5%) were the most common mutated genes. Patients with stable disease (SD)/progressive disease (PD) exhibited enrichment in cell cycle, FoxO, and TNF signaling pathways. A total of 29 patients underwent end-of-treatment PET/CT (EOT PET/CT), and 16 of them received ctDNA monitoring with 12 negative. Among 6 patients with EOT PET/CT positive (Deauville 4), 4 underwent ctDNA monitoring, and 3 of them were negative, being still in continuous remission without any subsequent anti-tumor therapy.
CONCLUSION
CtDNA may be combined with PET/CT to assess efficacy, monitor relapse, and guide treatment of PMBCL.
Humans
;
Circulating Tumor DNA/blood*
;
Female
;
Mediastinal Neoplasms
;
Male
;
Retrospective Studies
;
High-Throughput Nucleotide Sequencing
;
Prognosis
;
Lymphoma, Large B-Cell, Diffuse/genetics*
;
Middle Aged
;
Adult
;
Aged
;
Neoplasm Recurrence, Local
;
Mutation
7.Effect of Daratumumab, Lenalidomide, and Dexamethasone on Quality of Life in Patients with Newly Diagnosed Multiple Myeloma Ineligible for Stem Cell Transplantation.
Zhi-Hui LI ; Jin-Hui WANG ; Meng-Meng LIU ; Peng-Tao XING ; Yan-Ping ZHANG ; Xin-Rong ZHAN
Journal of Experimental Hematology 2025;33(3):816-821
OBJECTIVE:
To investigate the effect of daratumumab, lenalidomide and dexamethasone on quality of life in transplant-ineligible (TIE) patients with newly diagnosed multiple myeloma (NDMM).
METHODS:
The clinical data of 93 TIE NDMM patients in our hospital from January 2020 to December 2022 were retrospectively analyzed. The patients were divided into D-Rd group (48 cases) and Rd group (45 cases) according to treatment regimen. The patients in Rd group were treated with lenalidomide and dexamethasone, while those in D-Rd group were treated with daratumumab on the basis of Rd group. The QLQ-C30 and EQ-5D VAS scores of the two groups were compared at baseline and after 3, 6 and 12 treatment cycles. The last follow-up date was June 30, 2023, and overall survival (OS) was compared between the two groups.
RESULTS:
The median follow-up period in the D-Rd group was 21 (7-38) months, and the median OS was 34 months, while that in the Rd group was 16 (5-35) months, and the median OS was 28 months. There was significant difference in OS between the two groups ( P <0.05). After 3, 6 and 12 treatment cycles, the QLQ-C30 score and EQ-5D VAS score of the two groups were significantly improved (all P <0.05). After 3 and 12 treatment cycles, the QLQ-C30 score and EQ-5D VAS score of D-Rd group were significantly higher than those of Rd group (all P <0.05). There were no significant differences in the improvement of QLQ-C30 GHS and pain scores between the two groups of patients with age <75 years and ECOG 0-1 score after 3, 6 and 12 treatment cycles (P >0.05). In D-Rd group of patients with age≥75 years, the improvement of QLQ-C30 GHS scores after 3 and 12 treatment cycles and QLQ-C30 pain scores after 3, 6 and 12 treatment cycles was significantly superior to that in Rd group (all P <0.05). In D-Rd group of patients with ECOG 2 scores, the improvement of QLQ-C30 GHS and pain scores after 3, 6 and 12 treatment cycles was significantly superior to that in Rd group (all P <0.05).
CONCLUSION
Daratumumab, lenalidomide, and dexamethasone can significantly improve OS in TIE NDMM patients without decrease of quality of life, especially in those with age≥75 years or ECOG 2 scores.
Humans
;
Multiple Myeloma/drug therapy*
;
Lenalidomide/therapeutic use*
;
Quality of Life
;
Dexamethasone/therapeutic use*
;
Retrospective Studies
;
Antibodies, Monoclonal/therapeutic use*
;
Female
;
Male
;
Middle Aged
;
Aged
;
Stem Cell Transplantation
8.Protective Effect against Helicobacter pylor Gastritis in Mice by Flavonoid Combinations of Alpiniae Officinarum Rhizoma via Inhibition of PI3K/Akt Pathway
Xin LUO ; Wuyinxiao ZHENG ; Jingyu YANG ; Jianting ZHAN ; Haoran MA ; Xiaochuan YE ; Guopin GAN ; Dan LIU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(8):61-68
ObjectiveTo investigate the protective effect and mechanism of action of flavonoid combination of Alpiniae Officinarum Rhizoma (A. officinarum) against Helicobacter pylori (H. pylori) gastritis in mice. MethodsAfter acclimatization for one week, 56 SPF-grade healthy C57BL/6J mice were gavaged with mixed antibiotics for three consecutive days. They were randomly divided into a normal group, model group, positive drug group (triple therapy group), and low- and high-dose groups (100, 200 mg·kg-1) of flavonoid combination of A. officinarum. The H. pylori gastritis mice model was established by gavage with H. pylori bacterial suspension in each group except for the normal group. After successful modeling, mice were administrated with corresponding drugs once a day for two weeks. Hematoxylin-eosin (HE) staining was used to observe the pathological changes in gastric tissue. Rapid urease test paper was used to detect the positive rate of H. pylori. Silver staining was used to observe the H. pylori adherence on the surface of gastric tissue. Immunohistochemistry was used to detect the protein expression of interleukin-8 (IL)-8 and myeloid differentiation factor (MyD88) in gastric tissue. The serum levels of IL-6, tumor necrosis factor-α (TNF-α), IL-8, and IL-1β were detected by enzyme-linked immunosorbent assay (ELISA). The expressions of phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt) protein were detected by Western blot. ResultsCompared with those in the normal group, mice in the model group had lower gastric weight coefficients, higher pH of gastric juice, 100% H. pylori infection rate, and significantly changed gastric histopathology. The expressions of IL-8 and MyD88 proteins in the gastric tissue of mice in the model group were significantly elevated, and the serum levels of inflammatory factors IL-6, TNF-α, IL-8, and IL-1β were significantly up-regulated in mice. Compared with that in the model group, the gastric weight coefficient of mice in each treatment group of the flavonoid combinations of A. officinarum was elevated (P<0.01), and the pH of gastric juice was reduced (P<0.01). The infection rate of H. pylori was reduced. The expressions of IL-8 and MyD88 proteins in the gastric tissue of mice in the treatment groups were significantly reduced (P<0.01), and the serum levels of inflammatory factors IL-6, TNF-α, IL-8, and IL-1β were significantly reduced in a dose-dependent manner (P<0.01). The flavonoid combinations of A. officinarum down-regulated the expression of PI3K and Akt proteins in H. pylori gastritis-infected cells (P<0.01). ConclusionThe protective effect of flavonoid combinations of A. officinarum against H. pylori gastritis is associated with the inhibition of H. pylori infection rate and regulation of PI3K/Akt signaling pathway, resulting in inhibiting the release of inflammatory factors.
9.A questionnaire survey and analysis on the current situation of forensic ethics practice and educational needs
Wenjie LUO ; Tiantian PAN ; Shiyue LI ; Mengjun ZHAN ; Lirong QIU ; Yuchi ZHOU ; Xin CHEN ; Fei FAN ; Zhenhua DENG
Chinese Medical Ethics 2025;38(3):378-384
ObjectiveTo explore the current situation of forensic ethics practice and education by designing a questionnaire on forensic ethics, with a view to exploring the path of forensic ethics education construction. MethodsA total of 667 valid questionnaires were collected using the online survey method, basically covering various regions across the country and all sub-specialties of forensic medicine. Descriptive analysis was used to analyze the relevant data. ResultsMost practitioners had relevant ethical reflections in the process of forensic practice. 69.12% of the respondents indicated that they had studied the relevant rules, but approximately half stated that there were no corresponding ethical norms or standard operating manuals. The specific behaviors violating ethics in different units were diverse. 23.04% of the respondents reported that they had encountered unethical behaviors, but only 4.9% of them reported such violations. In terms of forensic ethics education, 87.75% of the respondents believed that there were issues with the current model of forensic ethics education. Meanwhile, the respondents showed a high degree of recognition for receiving forensic ethics education, with 84.15% of respondents expressing willingness to participate in relevant courses. More than half of respondents were willing to participate in forensic ethics education during undergraduate studies, new employee training, and regular post-employment training. ConclusionCurrently, there is a problem of ethical neglect in forensic work in China. Combining ethics courses with professional courses at the practitioner training stage and providing regular training at the practice stage are effective measures to popularize forensic ethics knowledge, enhance ethical awareness, and improve the quality of practice.
10.Mitochondrial Function and Regulation in Spermatogenesis and Activation of Caenorhabditis elegans
Zhan-Xin CHANG ; Long MIAO ; Peng WANG
Progress in Biochemistry and Biophysics 2025;52(7):1661-1672
Mitochondria play a pivotal role in spermatogenesis and sperm activation in Caenorhabditis elegans, serving as the primary ATP supplier for cell division and differentiation while also acting as a key regulator of zinc ion homeostasis, membrane dynamics, and apoptotic signaling. This review systematically summarizes the essential mitochondrial mechanisms at different stages of sperm development, highlighting their multifaceted contributions beyond energy metabolism. Mitochondria are crucial for maintaining the health and stability of the gonads by regulating key apoptotic execution proteins that facilitate the proper elimination of damaged or unnecessary germ cells. Additionally, mitochondria dynamically adjust their energy supply to meet the metabolic demands of different stages of germline development. During early spermatogenesis, mitochondria provide ATP to fuel mitotic and meiotic divisions, support cellular differentiation, and regulate H+ and Zn2+ exchange to maintain cytoplasmic homeostasis, thereby ensuring the proper maturation and functionality of sperm cells. As spermatogenesis progresses, mitochondria participate in processing and sorting essential sperm proteins, such as major sperm protein (MSP), and contribute to the formation of membranous organelles (MOs), which are critical for subsequent activation events. During sperm activation, mitochondria play a dual role in ensuring a successful transition from immotile spermatids to fully functional spermatozoa. First, they provide ATP to facilitate pseudopod formation, MO fusion, and ion channel regulation, all of which are essential for sperm motility and fertilization potential. Second, mitochondria regulate the quality and quantity of functional mitochondria within sperm cells through mitopherogenesis—a recently discovered process in which mitochondrial vesicles are selectively released, ensuring that only healthy mitochondria are retained. This quality-control mechanism optimizes mitochondrial function, which is crucial for sustaining sperm motility and longevity. Beyond their traditional role in energy metabolism, mitochondria may also contribute to protein synthesis during spermatogenesis and activation. Recent evidence suggests that mitochondrial ribosomes actively translate specific proteins required for sperm function, challenging the long-standing belief that spermatozoa do not engage in de novo protein synthesis after differentiation. This emerging perspective raises important questions about the role of mitochondria in regulating sperm activation at the molecular level, particularly in modulating oxidative phosphorylation (OXPHOS) protein composition to optimize ATP production. In summary, mitochondria serve as both the central energy hub and a crucial regulatory factor in sperm activation, metabolic homeostasis, and reproductive success. Their involvement extends beyond ATP generation to include apoptotic regulation, ion homeostasis, vesicle-mediated mitochondrial quality control, and potential contributions to protein synthesis. Understanding these mitochondrial functions in C. elegans not only deepens our knowledge of nematode reproductive biology, but also provides valuable insights into broader mechanisms governing mitochondrial regulation in germline cells across species. These findings open new avenues for future research into the interplay between mitochondria, energy metabolism, and sperm function, with potential implications for reproductive health and fertility studies.

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