Genomics, Proteomics & Bioinformatics•2024•DOI: 10.1093/gpbjnl/qzad002
The order Acipenseriformes, which includes sturgeons and paddlefishes, represents “living fossils” with complex genomes that are good models for understanding whole-genome duplication (WGD) and ploidy evolution in fishes. Here, we sequenced and assembled the first high-quality chromosome-level genome for the complex octoploid Acipenser sinensis (Chinese sturgeon), a critically endangered species that also represents a poorly understood ploidy group in Acipenseriformes. Our results show that A. sinensis is a complex autooctoploid species containing four kinds of octovalents (8n), a hexavalent (6n), two tetravalents (4n), and a divalent (2n). An analysis taking into account delayed rediploidization reveals that the octoploid genome composition of Chinese sturgeon results from two rounds of homologous WGDs, and further provides insights into the timing of its ploidy evolution. This study provides the first octoploid genome resource of Acipenseriformes for understanding ploidy compositions and evolutionary trajectories of polyploid fishes.
Stem Cell Research & Therapy•2025•DOI: 10.1186/s13287-025-04621-x
Background Mesenchymal stem cells (MSCs) isolated from umbilical cord blood (UCB) exhibit significant therapeutic efficacy in endometriosis; however, the molecular mechanisms governing their regulation remain incompletely elucidated. This study delves into the regulatory functions of miR-210-3p and miR-31-5p, which are secreted via exosomes from hypoxia-damaged endometrial epithelial cells, in modulating the behavior of UCB-MSCs. Methods UCB-MSCs were transfected with specific inhibitors targeting miR-210-3p and miR-31-5p. Proliferation and migratory capacities were quantified using CCK8, EdU incorporation, Transwell, and scratch wound healing assays. Western blotting was employed to assess the expression of endometrial epithelial markers (CD9 and CK19) and stromal markers (Vimentin and CD13), alongside the phosphorylation status of JAK2 and STAT3. Dual-luciferase reporter assays were conducted to validate SDF2 and FGF7 as direct targets of miR-210-3p and miR-31-5p, respectively. Results Suppression of miR-210-3p and miR-31-5p significantly augmented the proliferative and migratory abilities of UCB-MSCs, while simultaneously enhancing their differentiation into endometrial epithelial cells and attenuating their transition into stromal cells. Concurrently, the phosphorylation levels of JAK2 and STAT3 were markedly elevated. Overexpression of SDF2 and FGF7 further amplified the proliferative, migratory, and epithelial differentiation capacities of UCB-MSCs, accompanied by heightened activation of the JAK2/STAT3 signaling pathway. Notably, SDF2 overexpression and FGF7 overexpression effectively counteracted the inhibitory effects exerted by miR-210-3p and miR-31-5p mimics on UCB-MSC proliferation, migration, and epithelial differentiation, mediated through the modulation of JAK2/STAT3 signaling. Conclusion miR-210-3p and miR-31-5p orchestrate the functional dynamics of UCB-MSCs by targeting SDF2 and FGF7, respectively, through the JAK2/STAT3 pathway. These findings unveil novel mechanistic insights into the regenerative potential of UCB-MSCs, offering promising avenues for therapeutic advancements in endometriosis.
Stem Cell Research & Therapy•2025•DOI: 10.1186/s13287-025-04378-3
Background Myocardial infarction (MI) is a severe disease that often associated with impaired angiogenesis and increased myocardial apoptosis. Mesenchymal stromal cells (MSCs) have been a promising candidate for treating myocardial infarction. However, functional heterogeneity of MSCs leads to inconsistent therapeutic efficiency and the current MSCs-based therapy lacks the concept and implementation of precision medicine. In this study, we compared the cardioprotective effect of UCMSCs and ADMSCs targeting the angiogenesis in a mouse MI model and screened out optimum MSCs candidate for precise clinical application. Methods The gene expression profiles of UCMSCs and ADMSCs were investigated through RNA sequencing analysis. To compare their angiogenic potential, we performed tube formation assay, Matrigel plug assays, and aortic ring assay, and analyzed pro-angiogenic genes via qPCR. Subsequently, UCMSCs and ADMSCs were respectively injected into myocardium after MI surgery in mice. On day 28 post-MI, echocardiography was performed to assess cardiac function. Histological analysis was performed to assess MSCs retention, angiogenesis, and myocardial apoptosis. Additionally, the anti-apoptosis effects mediated by MSCs were further evaluated using flow cytometry in hypoxia H9C2 and HL-1 cells. Results The RNA sequencing analysis revealed differences in gene expression related to angiogenesis and apoptosis pathways between UCMSCs and ADMSCs. UCMSCs presented greater pro-angiogenesis activity than ADMSCs in vitro and in vivo. Both of UCMSCs and ADMSCs improved cardiac function, decreased infarction area and inhibited cardiomyocyte apoptosis while promoting angiogenesis post-MI in mice. Notably, ADMSCs exerted a better cardioprotective function than UCMSCs and stronger anti-apoptotic effect on residual cardiomyocytes.
Stem Cell Research & Therapy•2024•DOI: 10.1186/s13287-024-03745-w
Background Biomaterials used in bone tissue engineering must fulfill the requirements of osteoconduction, osteoinduction, and osseointegration. However, biomaterials with good osteoconductive properties face several challenges, including inadequate vascularization, limited osteoinduction and barrier ability, as well as the potential to trigger immune and inflammatory responses. Therefore, there is an urgent need to develop guided bone regeneration membranes as a crucial component of tissue engineering strategies for repairing bone defects. Methods The mZIF-8/PLA membrane was prepared using electrospinning technology and simulated body fluid external mineralization method. Its ability to induce biomimetic mineralization was evaluated through TEM, EDS, XRD, FT-IR, zeta potential, and wettability techniques. The biocompatibility, osteoinduction properties, and osteo-immunomodulatory effects of the mZIF-8/PLA membrane were comprehensively evaluated by examining cell behaviors of surface-seeded BMSCs and macrophages, as well as the regulation of cellular genes and protein levels using PCR and WB. In vivo, the mZIF-8/PLA membrane's potential to promote bone regeneration and angiogenesis was assessed through Micro-CT and immunohistochemical staining. Results The mineralized deposition enhances hydrophilicity and cell compatibility of mZIF-8/PLA membrane. mZIF-8/PLA membrane promotes up-regulation of osteogenesis and angiogenesis related factors in BMSCs. Moreover, it induces the polarization of macrophages towards the M2 phenotype and modulates the local immune microenvironment. After 4-weeks of implantation, the mZIF-8/PLA membrane successfully bridges critical bone defects and almost completely repairs the defect area after 12-weeks, while significantly improving the strength and vascularization of new bone. Conclusions The mZIF-8/PLA membrane with dual osteoconductive and immunomodulatory abilities could pave new research paths for bone tissue engineering.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2026014
Common fragile sites (CFSs) are large genomic loci that are frequently deleted under replication stress and are thought to play a role in carcinogenesis as well as developmental disorders during early embryogenesis. They often appear as gaps or breaks in mitotic chromosomes by conventional optical microscopy and are also loci that are replicated in mitosis in a process called mitotic DNA synthesis (MiDAS). However, we still have a very poor understanding of the potential mechanisms underlying their genomic instability. We recently found that in normal mitotic chromosomes, there is a two-level hierarchy in the higher-order chromatin structure, with mechanically resistant ~90 nm Mitotic NanoDomains (MNDs) assembling into ~125 nm compact granules (CGs). In this work, we combine multiple super-resolution imaging techniques, including binding-activated localization microscopy (BALM), atomic force microscopy (AFM), and stochastic optical reconstruction microscopy (STORM), to characterize CFSs. Our super-resolution data indicate that sites that appear as gaps by conventional microscopy can be filled with chromatin of the size and shape of MNDs but not CGs. Moreover, we find that MiDAS loci only form an array of MNDs with no CGs. Taken together, our work suggests that under replication stress, CFS loci can not only be replicated but also assembled into higher-order chromatin. However, this organization is limited to the level of MNDs, which suggests that a failure to progress beyond MNDs to form CGs constitutes a key structural deficiency at these loci.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2025055
Platelets circulate in an inactive form in the body until they contact with defective areas of endothelial cells or encounter a clotting cascade [1]. Activated platelets release and express bioactive substances and acquire the ability to bind plasma fibrinogen. Abnormal activation of platelets is involved in atherosclerosis and thrombosis [2,3]. When platelets are stimulated and activated, phospholipase A2 is activated at the same time, which then cleaves membrane phospholipids and frees arachidonic acid (AA) [4]. The latter catalyzes thromboxane A2 (TXA2) via thromboxane synthetase [5], which is induced by the cyclooxygenase COX-1 to produce prostaglandins G2 and H2 [6]. TXA2 is highly unstable, with a half-life of only 30 s, and it is rapidly hydrolyzed to relatively stable thromboxane B2 (TXB2), which is then converted in the liver to 11-dehydrothromboxane B2 (11dH-TXB2), which has a longer half-life and is excreted in the urine [7]. Dehydrothromboxane B2 is the final stable metabolite of thromboxane A2, which is derived only from arachidonic acid metabolism and can represent thromboxane A2 level in the body [8]. Specifically, by inhibiting the action of COX-1, the most important enzyme in the process of arachidonic acid metabolism, aspirin inhibits the production of thromboxane A2, that is, the concentration of TXA2 affects the effect of aspirin on platelet aggregation. However, the half-life of TXA2 (including the metabolic intermediate TXB2) is too short to be accurately measured, so the detection of its metabolic end product 11dH-TXB2 can very accurately reflect the sensitivity of the body to aspirin [7]. The concentration of 11dH-TXB2 in the serum correlates well with the concentration of 11dH-TXB2 in the urine, so the determination of 11dH-TXB2 in the urine can more effectively reflect the production of TXA2 in vivo [8]. The 11dH-TXB2 concentration needs to be corrected with the urinary creatinine concentration to rule out the effects of the urine concentration and renal function, so random urine samples can be used for testing [6]. Aspirin can acetylate serine at the key site of cyclooxygenase and thus irreversibly inhibits the activity of COX-1, reduces the synthesis of TXA2, and blocks the production of TXA2 and its induced platelet aggregation. Low-dose aspirin (30–75 mg/day) can effectively inhibit 95% of COX-1 activity [7]. Since the production of TXA2 in serum is largely dependent on platelet COX-1 (a therapeutic target of aspirin), 11dH-TXB2 can be used as a monitor for aspirin-induced platelet inhibition [3]. Creatinine is a metabolic byproduct of muscle metabolism that is primarily excreted via glomerular filtration, and its level is indicative of renal function [9]. The 24-h creatinine clearance can also be used to determine the integrity of the sample or to correct the urine sample concentration with the creatinine ratio [10]. Currently, creatinine detection methods include the Jaffe method, enzymolysis spectrophotometry, HPLC, capillary electrophoresis, capillary zone electrophoresis, gas chromatography tandem mass spectrometry (GC-MS) and liquid chromatography tandem mass spectrometry (LC-MS/MS) [11]. At present, there is no method for the simultaneous detection of 11dH-TXB2 and creatinine. When both analytes are needed, separate tests must be performed, increasing the workload and sample volume requirements. Thus, developing a method that enables the concurrent quantification of 11dH-TXB2 and creatinine in a single assay remains a critical challenge. The aim of this study was to provide a method for the simultaneous detection of 11dH-TXB2 and creatinine and to alleviate the problem that 11dH-TXB2 and creatinine cannot be simultaneously detected. By developing a standardized quantitative approach for measuring 11dHTXB2 and creatinine in human urine, this study aims to provide reliable concentration data, thereby facilitating further clinical research and methodology optimization.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2025234
Bivalent chromatin maintains genes in low-expression, poised states in embryonic stem cells (ESCs). However, bivalent promoters correlate with the transcriptional activation of oncogenic programs in malignancies, a seemingly contradiction that remains to be resolved. Here, we identify a class of cancer-specific bivalent promoters (CSBPs) through the integration of a system-level longitudinal framework. Compared with ESCs, CSBPs are characterized by lower and narrower H3K27me3 deposition alongside abundant H3K4me3, thus permitting the persistent expression of genes critical for cancer stem cell (CSC) formation and maintenance, as exemplified by SOX9. The generation of CSBPs is essentially induced by the acquisition of H3K27me3 during cell state transition, which is mediated by specific binding of PRC2.1 and the de novo recruitment of PRC2.2. Notably, disrupting the bivalency of CSBPs significantly increases H3K4me3 levels, leading to hyperactivation of CSBPs and eventually inhibiting clonal expansion of CSCs and impairing tumorigenesis. Our study not only helps explain the puzzle of transcriptionally active bivalent genes in cancer but also provides insights into the development of therapies targeting phenotypic plasticity.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2025150
Rapid identification of pathogens responsible for bloodstream infection is critical for early intervention and effective treatment. Reporter phages, which are known for their exceptional sensitivity and specificity in pathogen detection, have garnered significant interest. In this study, we systematically evaluate phage genome editing strategies that combine homologous recombination with the CRISPR-Cas9 system. We investigate the impacts of homologous arm length, sgRNA activity, target site, and plasmid interactions on editing efficiency. Our results demonstrate that successful genome editing depends on both sufficient cleavage pressure and optimal homologous arm length, particularly when using low-activity sgRNAs. On the basis of these findings, we develop a highly efficient gene editing strategy TPMSR (triple-plasmid-mediated synchronous recombination) that overcomes the limitations of conventional methods that rely on high-activity sgRNA and restricted editing sites. Using the TPMSR strategy, we integrate the Nluc gene into phage T7, generating the reporter phage T7::Nluc, which is then incorporated into a microfluidic chip. Validation with 51 clinical isolates demonstrates outstanding sensitivity, specificity, and accuracy in detecting Escherichia coli in blood within 1.5 h at concentrations less than 30 CFU/mL. This study presents a robust strategy for phage genome engineering and develops a promising method for the rapid diagnosis of bloodstream infections caused by E. coli.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2025086
Liver regeneration is a critical adaptive response to hepatic injury, requiring precise metabolic reprogramming to meet the energetic and biosynthetic demands of proliferating hepatocytes. While the oncoprotein Gankyrin is well-established as a promoter of liver fibrosis and hepatocarcinogenesis, its role in metabolic adaptations underlying liver regeneration remains unclear. In this study, we demonstrate that Gankyrin deficiency in the liver (Gank△Hep/Y) induces hepatic hypertrophy and aberrant glycogen accumulation. Gankyrin expression is significantly upregulated after partial hepatectomy (PHx), whereas Gank△Hep/Y -PHx mice exhibit impaired liver regeneration. This impairment is marked by a delayed restoration of the liver-to-body weight ratio, blunted glycogenolysis, and reduced fatty acid uptake. Mechanistically, Gankyrin activates Pygl and Cd36, key regulators of glycogenolysis and lipid uptake, respectively. Pharmacological inhibition of PYGL activity retards liver regeneration. Furthermore, we identify a novel interaction between Gankyrin and FOXO1, wherein Gankyrin promotes FOXO1 ubiquitination and subsequent proteasomal degradation. This Gankyrin-dependent suppression of FOXO1 leads to the transcriptional upregulation of Pygl and Cd36, thereby fueling hepatocyte proliferation. Collectively, our findings reveal Gankyrin as a master regulator of liver regeneration, integrating metabolic reprogramming with proliferative signaling through the FOXO1-PYGL/CD36 axis. These insights not only elucidate the mechanistic underpinnings of liver regeneration but also unveil the therapeutic potential of targeting the Gankyrin/FOXO1 pathway to mitigate hepatic insufficiency and enhance regenerative capacity in clinical settings.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2025032
Cisplatin resistance is a major cause of poor prognosis in patients with cervical cancer. Dysregulation of long noncoding RNAs (lncRNAs) plays a key role in chemoresistance. Our results reveal that the lncRNA UCA1 is upregulated in cisplatin (DDP)-resistant cervical cancer tissues and HeLa cells. Mechanistically, the lncRNA UCA1 acts as a sponge for miR-195-5p, targeting IKBKB. UCA1 enhances proliferation, migration, and invasion while reducing apoptosis in DDP-resistant HeLa cells via the miR-195-5p/IKBKB axis. Additionally, UCA1 upregulates BNIP3Δex2 and p-p65 expressions and downregulates BNIP3 expression in DDP-resistant HeLa cells. Abnormal expressions of BNIP3Δex2 and BNIP3 significantly alter the malignant progression of HeLa/DPP cells. In vivo, UCA1 silencing inhibits growth, enhances apoptosis, and upregulates IKBKB, BNIP3Δex2, and p-p65 expressions while downregulating BNIP3 expression in subcutaneous xenografts in nude mice by targeting miR-195-5p. Overall, this study highlights a novel promising target for the treatment of DDP-resistant cervical cancer.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2024174
Berberine (BBR) is used to treat diarrhea clinically. However, its reproductive toxicity is unclear. This study aims to investigate the impact of BBR on the male reproductive system. Intragastric BBR administration for 14 consecutive days results in a significant decrease in the serum testosterone concentration, epididymal sperm concentration, mating rate and fecundity of male mice. Testicular treatment with testosterone propionate (TP) partially reverses the damage caused by BBR to the male reproductive system. Mechanistically, the decrease in Muribaculaceae abundance in the gut microbiota of mice is the principal cause of the BBR-induced decrease in the sperm concentration. Both fecal microbiota transplantation (FMT) and polyethylene glycol (PEG) treatment demonstrate that Muribaculaceae is necessary for spermatogenesis. The intragastric administration of Muribaculaceae intestinale to BBR-treated mice restores the sperm concentration and testosterone levels. Metabolomic analysis reveals that BBR affects arginine and proline metabolism, of which ornithine level is downregulated. Combined analysis via 16S rRNA metagenomics sequencing and metabolomics shows that Muribaculaceae regulates ornithine level. The transcriptomic results of the testes indicate that the expressions of genes related to the low-density lipoprotein receptor (LDLR)-mediated testosterone synthesis pathway decrease after BBR administration. The transcriptional activity of the Ldlr gene in TM3 cells is increased with increased ornithine supplementation in the culture media, leading to increased testosterone synthesis. Overall, this study reveals an association between a BBR-induced decrease in Muribaculaceae abundance and defective spermatogenesis, providing a prospective therapeutic approach for addressing infertility-related decreases in serum testosterone triggered by changes in the gut microbiota composition.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2025139
Sepsis-associated acute liver injury (SALI) is a frequent and clinically severe complication of sepsis, in which inflammatory responses and oxidative stress are involved. Angelicin (ANG), one of the main active components in the traditional Chinese medicine Psoralea corylifolia Linn., has anti-inflammatory and antioxidant bioactivities. In this study, the protective effect of ANG on SALI and its specific mechanism are investigated by establishing a mouse model of caecal ligation and puncture (CLP)-induced SALI and an in vitro sepsis model in LPS-stimulated AML12 cells. These results show that ANG can alleviate liver injury and improve liver function in SALI mice. ANG decreases the mRNA expression levels of the pro-inflammatory factors Il-1β, Il-6, and Tnf-α and increases the mRNA expression level of the anti-inflammatory factor Il-10, which suggests its anti-inflammatory effects. The results of the biochemical kit assay and DHE staining show that ANG can decrease the levels of MDA and ROS and increase the level of GSH and the activities of CAT and SOD, which suggests that ANG has antioxidant effects. Mechanistically, ANG exerts anti-inflammatory effects by inhibiting the NF-κB and p38 MAPK pathways and exerting antioxidant effects by activating the Nrf2/Keap1 pathway. Additionally, cell transfection experiments indicate that activation of the Nrf2/Keap1 pathway by ANG may depend on the inhibition of the NF-κB pathway. In conclusion, ANG attenuates SALI by inhibiting the NF-κB and p38 MAPK pathways, thereby activating the Nrf2/Keap1 pathway and making it a promising therapeutic intervention for SALI.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2026014
Common fragile sites (CFSs) are large genomic loci that are frequently deleted under replication stress and are thought to play a role in carcinogenesis as well as developmental disorders during early embryogenesis. They often appear as gaps or breaks in mitotic chromosomes by conventional optical microscopy and are also loci that are replicated in mitosis in a process called mitotic DNA synthesis (MiDAS). However, we still have a very poor understanding of the potential mechanisms underlying their genomic instability. We recently found that in normal mitotic chromosomes, there is a two-level hierarchy in the higher-order chromatin structure, with mechanically resistant ~90 nm Mitotic NanoDomains (MNDs) assembling into ~125 nm compact granules (CGs). In this work, we combine multiple super-resolution imaging techniques, including binding-activated localization microscopy (BALM), atomic force microscopy (AFM), and stochastic optical reconstruction microscopy (STORM), to characterize CFSs. Our super-resolution data indicate that sites that appear as gaps by conventional microscopy can be filled with chromatin of the size and shape of MNDs but not CGs. Moreover, we find that MiDAS loci only form an array of MNDs with no CGs. Taken together, our work suggests that under replication stress, CFS loci can not only be replicated but also assembled into higher-order chromatin. However, this organization is limited to the level of MNDs, which suggests that a failure to progress beyond MNDs to form CGs constitutes a key structural deficiency at these loci.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21236
BACKGROUND: Patients with lumbar disc herniation (LDH) often experience comorbid anxiety due to chronic pain and functional limitations, significantly affecting their quality of life. However, the mechanisms underlying the pain-anxiety comorbidity remain unclear. OBJECTIVE: To investigate the neural regulatory mechanisms of the paraventricular nucleus in the hypothalamus in a mouse model of lumbar disc herniation with chronic pain-anxiety comorbidity. METHODS: A total of 100 C57BL/6 mice were randomly divided into a normal group (24 mice) and a model group (76 mice). The lumbar disc herniation model was established in the model group using a needle puncture method. Seventy-two successfully modeled mice were randomly divided into the model group, oxytocin group, and oxytocin+Vasotocin group, with 24 mice in each group. Mice in the oxytocin group received a 200 nL injection of oxytocin (0.5 μg/μL) into the paraventricular nucleus of the hypothalamus. Mice in the oxytocin+Vasotocin group received a 200 nL injection of oxytocin into the paraventricular nucleus and a 20 μL intraperitoneal injection of Vasotocin (an oxytocin antagonist, 0.15 μg/μL). Anxiety-like behavioral changes were evaluated via the elevated plus maze and open field tests on day 20 after modeling. Mechanical paw withdrawal threshold and thermal paw withdrawal latency experiments were conducted for all groups before modeling and 21 days after modeling. On day 21 post-modeling, immunofluorescence staining was used to observe c-FOS expression in the paraventricular nucleus of the hypothalamus; qPCR was used to detect mRNA expression of inflammatory factors prostaglandin E2, tumor necrosis factor α, and interleukin-1β in dorsal root ganglion tissue; Western blot was used to detect oxytocin receptor and p-ERK1/2 protein expression in the paraventricular nucleus. RESULTS AND CONCLUSION: Compared with the normal group, the model group showed significantly decreased mechanical and thermal pain thresholds (P < 0.05), significantly reduced time and entries in the open arms of the elevated plus maze (P < 0.05), significantly reduced time and entries in the open field (P < 0.05), significantly increased mRNA expression of prostaglandin E2, tumor necrosis factor α, and interleukin-1β in dorsal root ganglion tissue (P < 0.05), significantly increased c-FOS expression in the paraventricular nucleus (P < 0.05), significantly decreased oxytocin receptor protein expression, and significantly increased p-ERK1/2 protein expression (P < 0.05). Compared with the model group, the oxytocin group showed significantly increased mechanical and thermal pain thresholds (P < 0.05), significantly increased time and entries in the open arms of the elevated plus maze (P < 0.05), significantly increased time and entries in the open field (P < 0.05), significantly decreased mRNA expression of prostaglandin E2, tumor necrosis factor α, and interleukin-1β in dorsal root ganglion tissue (P < 0.05), significantly decreased c-FOS expression in the paraventricular nucleus (P < 0.05), significantly increased oxytocin receptor protein expression, and significantly decreased p-ERK1/2 protein expression (P < 0.05). Compared with the oxytocin group, the use of Vasotocin reversed the beneficial effects of oxytocin on pain and anxiety, increased inflammatory factor expression, significantly decreased oxytocin receptor protein expression, and significantly increased p-ERK1/2 protein expression. These results indicate that oxytocin can significantly improve chronic pain and anxiety-like behavior in mice with lumbar disc herniation, inhibit dorsal root ganglion inflammation, and the mechanism may be related to activation of the ERK signaling pathway in the paraventricular nucleus and downregulation of inflammatory factor expression.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21339
BACKGROUND: Mixed lineage kinase domain-like protein is one of the key executor proteins in the necroptosis pathway and plays an important role in various diseases. However, the mechanism by which its lactylation affects the formation and differentiation of breast tumor stem cells remains unclear. OBJECTIVE: To investigate the effect of mixed lineage kinase domain-like protein K230 site lactylation on the stemness expression of breast tumor stem cells. METHODS: The differences in protein lactylation between breast tumor MCF-7 adherent cells and spheroidal stem cells were analyzed by mass spectrometry. The mixed lineage kinase domain-like protein and its lactylation sites related to tumor stem cells were screened. A mixed lineage kinase domain-like protein K230R mutant plasmid vector was constructed and transfected into MCF-7 breast tumor cells. The proliferation and migration abilities of the cells were detected by CCK-8 and scratch assays. The effect of mixed lineage kinase domain-like protein K230R mutation on the formation of breast tumor stem cells was verified by suspension spheroid formation assay. Western blot was used to detect the expression of stemness and epithelial-mesenchymal transition-related proteins. RESULTS AND CONCLUSION: The lactylation level of mixed lineage kinase domain-like protein at K230 was higher in breast tumor stem cells. Compared with the wild-type group, the K230R mutant group showed significantly reduced scratch healing rate and spheroid formation rate, significantly increased expression of epithelial-related proteins, and significantly decreased expression of mesenchymal-related and stemness-related proteins. The study indicates that lactylation of mixed lineage kinase domain-like protein at K230 promotes epithelial-mesenchymal transition and enhances stemness expression of breast tumor stem cells, thereby promoting the occurrence and development of breast tumors.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21455
BACKGROUND: Targeting the molecular mechanisms of ferrostatin, intervening in iron metabolism or inhibiting lipid peroxidation is expected to be a new strategy for the treatment of lumbar disc herniation, providing a new research direction for disease prevention and treatment. OBJECTIVE: To investigate the mechanism of action of the ferroptosis inhibitor ferrostatin-1 on lumbar disc herniation through in vitro cell experiments and in vivo animal studies using poly(lactic-co-glycolic acid)-poly(ethylene glycol)-poly(lactic-co-glycolic acid) hydrogel as a carrier. METHODS: (1) Third-generation mouse nucleus pulposus cells were divided into three treatment groups: the control group received no treatment; the model group received 10 ng/mL interleukin-1β, and the ferrostatin-1 group received 10 ng/mL interleukin-1β plus 25 μmol/L ferrostatin-1. Intracellular malondialdehyde levels, glutathione levels, iron ion content, and the mRNA expression of extracellular matrix-related genes type II collagen, aggrecan, matrix metalloproteinase 3 were detected. (2) Ferrostatin-1-loaded poly(lactic-co-glycolic acid)-poly(ethylene glycol)-poly(lactic-co-glycolic acid) hydrogel (drug-loaded hydrogel) was prepared, and its microstructure and in vitro drug release were characterized. Eighty C57BL/6 mice were randomly divided into normal, model, free drug, and drug-loaded hydrogel groups (n=20 per group). Except for the normal group, the other three groups were established as L5/6 lumbar disc herniation models. The model, free drug, and drug-loaded hydrogel groups received perivertebral injections of PBS, ferrostatin-1 solution, and drug-loaded hydrogel, respectively. Mechanical and thermal pain thresholds were dynamically monitored. On day 7 after administration, nucleus pulposus tissues were harvested to detect inflammatory factors (tumor necrosis factor α and interleukin-1β), malondialdehyde, glutathione levels, and ferroptosis pathway-related genes glutathione peroxidase 4 and solute carrier family 7 member 11 mRNA expression. RESULTS AND CONCLUSION: (1) Compared with the control group, the model group showed increased intracellular malondialdehyde levels, iron ion accumulation, and matrix metalloproteinase 3 mRNA expression (P < 0.05), while glutathione levels and type II collagen and aggrecan mRNA expression decreased (P < 0.05). Compared with the model group, the ferrostatin-1 group showed decreased intracellular malondialdehyde levels, iron ion accumulation, and matrix metalloproteinase 3 mRNA expression (P < 0.05), while glutathione levels and type II collagen and aggrecan mRNA expression increased (P < 0.05). (2) Scanning electron microscopy showed that the drug-loaded hydrogel had a loose porous structure with vacuoles of varying sizes, and the hydrogel exhibited good sustained-release properties. Compared with the model group, both free drug and drug-loaded hydrogel groups showed pain relief, decreased inflammatory factors and malondialdehyde levels (P < 0.05), and increased glutathione levels and glutathione peroxidase 4 and solute carrier family 7 member 11 mRNA expression (P < 0.05), with the drug-loaded hydrogel showing stronger effects than the free drug. (3) These results indicate that ferrostatin-1 exerts a protective effect on nucleus pulposus cells by regulating oxidative stress and ferroptosis-related gene expression, thereby treating lumbar disc herniation in mice.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21624
BACKGROUND: Lower extremity deep vein thrombosis is a catastrophic complication after lumbar spine fusion, and previous studies have shown that some patients, such as those with prosthetic arthroplasty and abdominal surgeries, suffer from a combination of preoperative anemia, which predisposes them to postoperative deep vein thrombosis of the lower extremities. However, whether preoperative anemia increases the risk of lower extremity deep vein thrombosis after lumbar spine fusion is unclear. OBJECTIVE: To investigate the correlation between preoperative anemia and deep vein thrombosis of the lower extremities in patients after lumbar fusion. METHODS: The clinical data of 1 178 patients who underwent lumbar spinal fusion treatment admitted to Third Affiliated Hospital of Anhui Medical University from January 2020 to December 2023 were retrospectively analyzed. According to whether or not the patients developed lower extremity deep vein thrombosis after the operation, the patients were divided into the deep vein thrombosis group (ultrasound report suggestive of lower extremity deep vein thrombosis) and the non-deep vein thrombosis group (ultrasound report suggestive of no lower extremity deep vein thrombosis). The incidence of anemia was compared between the two groups, and risk factors for lower extremity deep vein thrombosis in lumbar fusion patients were determined by univariate analysis and multivariate logistic regression analysis. RESULTS AND CONCLUSION: Among 1 178 patients, there were 43 cases (3.7%) in the deep vein thrombosis group and 1 135 cases (96.4%) in the non-deep vein thrombosis group. The incidence of preoperative anemia in the deep vein thrombosis group was 32.6%, significantly higher than that in the non-deep vein thrombosis group (10.9%, P < 0.05). Univariate analysis showed significant differences between the two groups in preoperative hemoglobin (P < 0.001), preoperative red blood cell count (P=0.028), D-dimer positivity (P=0.029), hypertension (P=0.019), number of fused segments (P=0.023), anemia (P < 0.001), and blood transfusion (P=0.006) (P < 0.05). Multivariate analysis showed that preoperative anemia (OR=4.221, 95%CI: 1.198-14.802, P=0.025) and D-dimer positivity (OR=2.023, 95%CI: 1.065-3.844, P=0.031) were risk factors for lower extremity deep vein thrombosis after lumbar fusion. These findings suggest that the incidence of preoperative anemia is relatively high in patients undergoing elective lumbar fusion, and preoperative anemia is an independent risk factor for deep vein thrombosis after lumbar fusion. It is recommended that preoperative anemia should be actively managed and corrected before elective lumbar fusion to reduce the risk of lower extremity deep vein thrombosis.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2025032
Cisplatin (DDP) resistance remains a principal determinant of poor prognosis in cervical cancer, with five-year survival at 16.5% for recurrent or advanced metastatic disease. The long noncoding RNA UCA1 is upregulated in DDP-resistant cervical cancer tissues and HeLa cells. Mechanistically, UCA1 functions as a competing endogenous RNA for miR-195-5p, thereby derepressing IKBKB. UCA1 knockdown suppresses proliferation, migration, and invasion while increasing apoptosis in DDP-resistant HeLa cells through the miR-195-5p/IKBKB axis. UCA1 upregulates BNIP3Δex2 and p-p65 and downregulates BNIP3 in DDP-resistant HeLa cells; forced changes in BNIP3Δex2 and BNIP3 expression significantly alter malignant progression of HeLa/DDP cells. In subcutaneous xenografts in nude mice, UCA1 silencing inhibits tumor growth, enhances apoptosis, and upregulates IKBKB, BNIP3Δex2, and p-p65 while downregulating BNIP3 via miR-195-5p targeting. These findings identify UCA1 as a promising therapeutic target for DDP-resistant cervical cancer.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2025055
Thromboxane A2 (TXA2) is a labile eicosanoid with a half-life of 30 s, limiting direct quantification. Its stable urinary metabolite, 11-dehydrothromboxane B2 (11dH-TXB2), reflects in vivo TXA2 biosynthesis and aspirin-mediated COX-1 inhibition. Creatinine normalization is required to correct for urine concentration and renal function. This study establishes a quantitative liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for simultaneous determination of 11dH-TXB2 and creatinine in human urine. Sample pretreatment involves organic phase extraction, eliminating solid-phase extraction. The method was validated for linearity, accuracy, precision, and recovery. The linear range for 11dH-TXB2 was 0.1–50 ng/mL (r² = 0.99656) and for creatinine 10–5000 ng/mL (r² = 0.99950). Quality control accuracies ranged from 85.83% to 113.21%, with RSDs below 9.71%. Standard recoveries were 85–110%. The assay meets regulatory requirements for simultaneous quantification. This approach provides a reliable tool for monitoring aspirin response and investigating thromboxane-related pathologies.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2025125
African swine fever virus (ASFV), a large DNA virus with a 170–193 kb genome encoding 150–167 proteins, including over 68 structural proteins, poses a significant threat to the global swine industry. The lack of a comprehensive protein library hinders functional, antigenic, and vaccine studies. Here, we constructed an ASFV proteome library by expressing recombinant ASFV proteins with an N-terminal glutathione S-transferase (GST) tag in Saccharomyces cerevisiae. Through optimization of codons, expression vectors, strains, and expression and purification conditions, we achieved satisfactory yields for analytical applications, covering approximately 95% of the ASFV proteome. A protein chip fabricated from 94 proteins with concentrations above 5 μg/mL and purities greater than 90% was used to screen interactions between ASFV and swine proteins (IRF3, p65, and IκBα). Among purification methods, 96-deep-well plate purification yielded the highest protein purity, outperforming gravity-flow and peristaltic-pump-flow column chromatography, as explained by chromatographic plate theory. This library and the optimized methods provide a foundation for understanding ASFV biology and for developing diagnostics and vaccines, and are instructive for generating other proteome libraries for high-throughput applications.