1.Effect of integrin α5 on NLRP3 expression in periodontal ligament fibroblasts within an inflammatory microenvironment
DAI Jingyi ; CAI Hongxuan ; SI Weixing ; ZHANG Zan ; WANG Zhurui ; LI Mengsen ; TIAN Ya guang
Journal of Prevention and Treatment for Stomatological Diseases 2025;33(1):24-32
Objective:
To investigate the effect of integrin α5 on the expression of NOD-like receptor thermal protein domain associated protein 3 (NLRP3) in periodontal ligament fibroblasts (PDLFs) within an inflammatory microenvironment.
Methods:
This study was approved by the Ethics Committee of Laboratory animals. After rat PDLFs were treated with LPS (0.5, 5, and 50 µg/mL) for 24 h, the primary medium was discarded and replaced with serum-free culture medium. After 24 h, the supernatant was collected and mixed with DMEM medium containing 10% exosome-free serum at a volume ratio of 1:1 to obtain conditioned medium (CM). The groups were labeled as the 0.5-CM, 5-CM, and 50-CM groups. In addition, PDLFs cultured in DMEM medium containing 10% exosome-free serum were considered the 0-CM group. PDLFs were cultured with the above CM. In the inhibitor group, PDLFs were cultured in 0-CM containing different concentrations of integrin α5 inhibitor ATN-161 (0, 0.025, 0.25, 2.5, 25, and 250 μg/mL). The effect of CM and integrin α5 inhibitor ATN-161 on cell viability was assessed using the CCK-8 assay. According to the CCK-8 results, in further inhibitor intervention experiments, PDLFs were cultured in 0-CM, 5-CM (without/with 25 μg/mL ATN-161), and 0-CM containing 25 μg/mL ATN-161, which were labeled as the 0-CM, 5-CM, ATN-161+5-CM, and ATN-161 groups, respectively. The expression changes of integrin α5 and NLRP3 were detected using Western blot and qRT-PCR techniques. For in vivo experiments, 24 rats were randomly divided into four groups (n=6). The control group contained healthy rats that received no treatment. The rats in the other three groups were injected with 40 µL of 0-CM containing 25 μg/mL ATN-161 or 5-CM (without or with 25 μg/mL ATN-161) on the palatal side of the left maxillary first molar every three days; these groups were classified as the ATN-161, 5-CM, and ATN-161+5-CM groups, respectively. On the 30th day, the left maxillary tissue of rats was used for Micro-CT, HE staining, and immunohistochemical detection.
Results :
The CCK-8 assay showed that CM, 25 μg/mL ATN-161, and ATN-161 concentrations below 25 μg/mL had no significant effect on cell viability at 12 h and 24 h (P > 0.05). 50-CM and 25 μg/mL ATN-161 significantly inhibited cell viability at 48 h (P < 0.05). For in vitro experiments, compared to the 0-CM group, both the protein and mRNA levels of integrin α5 and NLRP3 were significantly increased in rat PDLFs in the 5-CM group (P < 0.05). Intervention with 25 μg/mL ATN-161 significantly attenuated the enhancement of 5-CM on the expression of integrin α5 and NLRP3 (P < 0.05). For in vivo experiments, compared to the control group, alveolar bone resorption and periodontal inflammatory cell infiltration were significantly increased in the 5-CM and ATN-161+5-CM groups, and the expression of integrin α5 and NLRP3 was significantly increased (P < 0.01). However, compared to the 5-CM group, the ATN-161+5-CM group had less alveolar bone resorption and fewer periodontal inflammatory cells. Further, the expression of integrin α5 and NLRP3 was significantly reduced (P < 0.01).
Conclusion
In vitro and in vivo experiments showed that integrin α5 mediated NLRP3 expression in PDLFs under an inflammatory microenvironment. ATN-161 inhibited the expression of integrin α5, thus significantly downregulating the expression of NLRP3, which plays a role in inhibiting inflammation.
2.Endomitosis: a new cell fate in the cell cycle leading to polyploidy in megakaryocytes and hepatocytes.
Qi-Hua HUA ; Xuechun ZHANG ; Ruifeng TIAN ; Zhigang SHE ; Zan HUANG
Journal of Zhejiang University. Science. B 2025;26(9):843-862
Megakaryocytes and hepatocytes are unique cells in mammals that undergo polyploidization through endomitosis in terminal differentiation. Many polyploidization regulators and underlying mechanisms have been reported, most of which are tightly coupled with development, organogenesis, and cell differentiation. However, the nature of endomitosis, which involves successful entry into and exit from mitosis without complete cytokinesis, has not yet been fully elucidated. We highlight that endomitosis is a new cell fate in the cell cycle, and tetraploidy is a critical stage at the bifurcation of cell fate decision. This review summarizes the recent research progress in this area and provides novel insights into how cells manipulate mitosis toward endomitosis. Endomitotic cells can evade the tetraploidy restrictions and proceed to multiple rounds of the cell cycle. This knowledge not only deepens our understanding of endomitosis as a fundamental biological process but also offers new perspectives on the physiological and pathophysiological implications of polyploidization.
Hepatocytes/physiology*
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Megakaryocytes/physiology*
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Humans
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Polyploidy
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Animals
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Cell Cycle/physiology*
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Cell Differentiation
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Mitosis/physiology*
3.Application of Cytb and 12S rRNA in wildlife species identification for forensic science
Dezhi JIANG ; Yaxin ZHANG ; Yu ZANG ; Maolei AN ; Zan ZHANG ; Chengcheng QIU ; Yaoheng JIANG ; Wei SONG ; Hong ZHAO ; Kun XIE ; Jiayi CHEN ; Riga WU
Chinese Journal of Forensic Medicine 2025;40(3):308-311,322
Objective To analyze and compare the efficacy of DNA barcode,i.e.,Cytochrome b(Cytb)and 12S ribosomal RNA(12S rRNA)gene sequences,in the species identification of wildlife.Methods DNA extraction,quantification,PCR amplification of Cytb and 12S rRNA gene fragments,Sanger sequencing,and sequence alignment analysis were performed on ten wildlife samples.Results Both gene fragments were successfully amplified in six samples,while Cytb alone was successfully amplified in 1 sample,and 12S rRNA alone in 3 samples.Sequence analysis indicated that Cytb enabled species-level identification for 6 samples(Gallinula chloropus,Streptopelia orientalis,Phasianus colchicus,Falco naumanni,Myiopsitta monachus and Lynx lynx)and genus-level identification for 1 sample(Lepus).In contrast,12S rRNA achieved species-level identificaggion for 8 samples(Gallinula chloropus,Lepus sinensis,Phasianus colchicus,Myiopsitta monachus,Muntiacus reevesi,Macaca mulatta and Lynx lynx),representing seven species,and genus-level identification for 1 sample(Falco).However,by combining Cytb and 12S rRNA,all samples could be identified to the species level.Conclusion When applying DNA barcodes to wildlife identification,the Cytb and 12S rRNA gene regions analyzed here can effectively identify common species such as Gallinula chloropus and Streptopelia orientalis,but face difficulties in distinguishing closely related species within the same genus.Therefore,when conducting wildlife species identification,it is recommended to use two or more DNA barcode markers.
4.FOXO3-engineered human mesenchymal stem cells efficiently enhance post-ischemic stroke functional rehabilitation.
Fangshuo ZHENG ; Jinghui LEI ; Zan HE ; Taixin NING ; Shuhui SUN ; Yusheng CAI ; Qian ZHAO ; Shuai MA ; Weiqi ZHANG ; Jing QU ; Guang-Hui LIU ; Si WANG
Protein & Cell 2025;16(5):365-373
5.Single-nucleus transcriptomics decodes the link between aging and lumbar disc herniation.
Min WANG ; Zan HE ; Anqi WANG ; Shuhui SUN ; Jiaming LI ; Feifei LIU ; Chunde LI ; Chengxian YANG ; Jinghui LEI ; Yan YU ; Shuai MA ; Si WANG ; Weiqi ZHANG ; Zhengrong YU ; Guang-Hui LIU ; Jing QU
Protein & Cell 2025;16(8):667-684
Lumbar disc (LD) herniation and aging are prevalent conditions that can result in substantial morbidity. This study aimed to clarify the mechanisms connecting the LD aging and herniation, particularly focusing on cellular senescence and molecular alterations in the nucleus pulposus (NP). We performed a detailed analysis of NP samples from a diverse cohort, including individuals of varying ages and those with diagnosed LD herniation. Our methodology combined histological assessments with single-nucleus RNA sequencing to identify phenotypic and molecular changes related to NP aging and herniation. We discovered that cellular senescence and a decrease in nucleus pulposus progenitor cells (NPPCs) are central to both processes. Additionally, we found an age-related increase in NFAT1 expression that promotes NPPC senescence and contributes to both aging and herniation of LD. This research offers fresh insights into LD aging and its associated pathologies, potentially guiding the development of new therapeutic strategies to target the root causes of LD herniation and aging.
Intervertebral Disc Displacement/metabolism*
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Humans
;
Aging/pathology*
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Nucleus Pulposus/pathology*
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Male
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Female
;
Transcriptome
;
Middle Aged
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Lumbar Vertebrae/pathology*
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Adult
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Cellular Senescence
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Stem Cells/pathology*
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Aged
;
Intervertebral Disc Degeneration/metabolism*
7.Strategy and technical points of harvesting different forms of free rectus abdominis muscle flap combined with free superficial inferior epigastric artery perforator flap for unilateral breast reconstruction
Ruqi GUO ; Tianyi ZHANG ; Dajiang SONG ; Zhiyuan WANG ; Xu LIU ; Zan LI
Chinese Journal of Medical Aesthetics and Cosmetology 2025;31(2):93-98
The free inferior epigastric artery perforator flap is currently a highly regarded autologous breast reconstruction technique. However, in clinical practice, the anatomical characteristics of the lower abdominal vasculature and surrounding tissue structures do not always permit ideal perforator flap harvesting. In many cases, it becomes necessary to preserve a larger rectus abdominis muscle cuff to ensure reliable blood supply to the flap through the perforating vessels. Compared with various forms of rectus abdominis myocutaneous flaps that all require incision of the anterior sheath and intramuscular vascular pedicle dissection, the superficial inferior epigastric artery (SIEA) perforator flap significantly reduces donor-site morbidity. Nevertheless, the anatomical variability of the superficial inferior epigastric vessels increases surgical uncertainty. Based on clinical observations, this study establishes a comprehensive clinical strategy algorithm. Primarily focusing on different configurations of free rectus abdominis flaps, it incorporates additional preparation of free SIEA perforator flaps when anatomical conditions permit. The objectives are to ensure reliable perfusion of the lower abdominal flap while minimizing donor-site damage and reducing surgical complexity.
8.Up-regulation of CircPDS5B level involving in the pathophysiological mechanism of premature birth
Zan-yi HUANG ; Mei-sha FU ; Hui-li CHEN ; Lin-jing ZHANG
Journal of Regional Anatomy and Operative Surgery 2025;34(8):668-674
Objective To explore the pathophysiological molecular mechanism of the up-regulation of CircPDS5B level involved in premature birth.Methods The placental tissues of full-term infants(Group 1,gestational age≥37 weeks),late premature infants(Group 2,34 weeks≤gestational age<37 weeks),premature infants(Group 3,32 weeks
9.Clinical effects of lobulated supercharged pedicled rectus abdominis myocutaneous flap for repairing huge chest wall wounds
Xinshan ZHANG ; Junyi YU ; Zan LI ; Dajiang SONG
Chinese Journal of Burns 2025;41(3):251-257
Objective:To explore the clinical effects of lobulated supercharged pedicled rectus abdominis myocutaneous flap for repairing huge chest wall wounds.Methods:This study was a retrospective observational study. From January 2020 to June 2023, 9 patients with huge chest wall wounds remained after extended radical tumor resection, extended tumor resection, or lesion resection and conformed to the inclusion criteria were admitted to Hunan Cancer Hospital, including 3 males and 6 females, aged 31-59 years. The lobulated supercharged pedicled rectus abdominis myocutaneous flap was used to repair the huge chest wall wounds. The area of chest wall skin and soft tissue defects ranged from 19 cm×15 cm to 25 cm×21 cm, and the area of the harvested myocutaneous flap ranged from 25.0 cm×7.5 cm to 32.0 cm×13.0 cm. After repairing the rectus abdominis muscle and its anterior sheath with a polypropylene mesh, the incision in the donor site was directly sutured. The blood supply of the myocutaneous flap and the selection of blood vessels in recipient area for supercharging during the surgery, the survival of the myocutaneous flap and the healing of the donor area incision after the surgery were observed. The appearance and texture of the reconstructed chest wall, the scar formation in the abdominal donor area, and their impacts on function and appearance, and the tumor recurrence and metastasis were followed up.Results:The blood supply of the unilateral myocutaneous flap was poor in 7 patients, and that of the bilateral myocutaneous flaps was poor in 2 patients during the surgery. The recipient area vessels selected for supercharging of the myocutaneous flap were the internal thoracic vessels in 7 patients, the thoracodorsal vessels in 2 patients, and the thoracoacromial vessels in 2 patients. All the myocutaneous flaps survived after surgery. The donor area incisions of 7 patients healed smoothly after surgery; 2 patients had partial dehiscence in the incisions due to excessive incision tension, which healed after debridement and suturing. Follow-up for 8 to 12 months showed that the reconstructed chest wall had good appearance and soft texture; only a linear scar remained in the abdominal donor area, which had no obvious impact on abdominal breathing and the abdomen was aesthetically pleasing; no local tumor recurrence was observed; distant metastases occurred in 2 breast cancer patients.Conclusions:The lobulated supercharged pedicled rectus abdominis myocutaneous flap can effectively cover huge chest wall wounds, while maximally ensuring the blood supply of the myocutaneous flap to the greatest extent and safeguarding the success of chest wall reconstruction surgery.
10.Contralateral posteromedial thigh flap for salvage breast reconstruction with adductor magnus perforator flap failure
Dajiang SONG ; Tianyi ZHANG ; Zan LI ; Yixin ZHANG
Chinese Journal of Plastic Surgery 2025;41(6):577-582
Objective:To summarize the technical experience of using a contralateral free gracilis myocutaneous flap combined with an adductor magnus perforator flap for salvaging failure of breast reconstruction after transplantation with the free adductor magnus perforator flap.Methods:A retrospective analysis was conducted on patients with breast cancer who received contralateral posteromedial thigh flap (gracilis myocutaneous flap combined with adductor magnus perforator flap) transplantation for immediate unilateral breast reconstruction developed vascular crisis and caused flap necrosis at the Department of Oncology Plastic Surgery, Hunan Cancer Hospital, between December 2016 to December 2022. All patients with early-stage breast cancer received modified radical mastectomy. All patients were selected to undergo the immediate breast reconstruction surgery with unilateral free adductor magnus perforator flap transplantation. The proximal end of internal mammary vessels were used as the recipient vessels in all cases. After the emergency exploration confirmed the necrosis of the flap, a contralateral free gracilis myocutaneous flap combined with adductor magnus perforator flap were immediately harvested for salvage breast reconstruction. The vascular pedicle of the gracilis muscular branch and the adductor magnus perforator branch were anastomosed with the proximal and distal ends of the internal mammary vessels, respectively. Postoperative patient monitoring was conducted intensively, with follow-up assessments focusing on breast contour and donor site recovery.Results:A total of 5 patients were included, aged 26 to 42 years, with an average of (31.5±1.8) years. All salvaged breast reconstruction surgeries were successful, the salvaged flap measured 15.0 cm×6.0 cm×4.0 cm-17.0 cm×7.5 cm×5.5 cm. All the transplanted flaps survived after the operation, with satisfactory breast contour, good flap elasticity, and no contracture or deformation. The donor sites of both thigh flaps healed well, leaving only linear scars and no significant functional impairment to the lower limbs. Follow-up ranged from 9 to 24 months, with an average of 15.7 months. The breast shape was satisfactory, and there was no recurrence of breast cancer.Conclusion:The donor site morbidity of the posteromedial thigh flap is relatively small. When unilateral flap transplantation fails, immediate transfer of the contralateral flap can be chosen for salvage reconstruction. This approach ensures symmetrical donor site outcomes and is generally well-accepted by patients.


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