ObjectiveTo optimize the extraction method for polysaccharides from turnip（Brassica rapa）， and analyze and evaluate the primary structure of the isolated and purified turnip polysaccharide fraction（BP-1） and its hypoglycemic effects in diabetic zebrafish. MethodsTaking polysaccharide yield as the evaluation index， a semi-bionic extraction method was employed. Single-factor experiments and Box-Behnken response surface methodology were used to investigate three factors of solid-to-liquid ratio， extraction time and extraction temperature， in order to optimize the extraction process. BP-1 was isolated and purified using the Sevage method and DEAE-52 cellulose column chromatography. Structural characterization of the turnip polysaccharides was performed using ultraviolet-visible spectrophotometry（UV）， gas chromatography-mass spectrometry（GC-MS）， Congo red assay， and Fourier-transform infrared spectroscopy（FT-IR） to determine purity， monosaccharide composition， triple-helix structure， and functional groups. The microstructure of the polysaccharides was observed using scanning electron microscopy（SEM） and atomic force microscopy（AFM）. Zebrafish were divided into the blank group（adding E3 medium）， and BP-1-1， BP-1-10， BP-1-50， BP-1-200， BP-1-1 000 groups（adding BP-1 solutions at concentrations of 1， 10， 50， 200， 1 000 mg·L^-1， respectively）， and zebrafish embryos were subjected to a 96-hour exposure experiment. The maximum tolerated concentration of BP-1 in zebrafish was determined by evaluating its effects on phenotype， survival rate， malformation rate， and heart rate. Experimental animals were randomly divided into the blank group， model group， BP-1-10 group（10 mg·L^-1）， BP-1-50 group（50 mg·L^-1）， and BP-1-200 group（200 mg·L^-1）. The blank group was cultured in E3 medium， the model and treatment groups were induced to establish a diabetic model in 4 day-post-fertilization（dpf） zebrafish embryos using 10 g·L^-1 of glucose combined with 500 µmol·L^-1 of alloxan. The treatment groups received corresponding doses of BP-1 solution， while the blank and model groups received an equal volume of saline. Glucose and insulin（INS） levels were measured using enzyme-linked immunosorbent assay（ELISA） kits， the effects on the liver were observed by hematoxylin-eosin（HE） histopathological sections. The mRNA expression levels of glucagon（Glucagon）， insulin（Insa）， and phosphoenolpyruvate carboxykinase 1（PCK1） were detected with real-time fluorescence quantitative polymerase chain reaction（Real-time PCR）. ResultsThe optimized extraction conditions were determined as follows：solid-to-liquid ratio of 1∶40（g·mL^-1）， extraction time of 66 min， and extraction temperature of 79 ℃. Under these conditions， the yield of turnip polysaccharides was （10.34±0.96）%. UV analysis indicated that BP-1 contained no proteins or nucleic acids， GC-MS analysis revealed that BP-1 consisted of six monosaccharides（arabinose， rhamnose， ribose， mannose， galactose and glucose）. Congo red assay indicated that the molecular conformation did not exhibit a triple-helix structure， FT-IR analysis showed the presence of α-glycosidic bonds and uronic acids， SEM analysis revealed an irregular flaky structure with a flat and smooth surface， AFM analysis suggested that the aggregated structure might be formed by the entanglement of molecular chains and intramolecular hydrogen bonding. The maximum tolerated concentration of BP-1 in zebrafish over 96 h was determined to be 200 mg·L^-1. Pharmacodynamic results showed that， compared with the blank group， the model group exhibited significantly increased glucose levels and significantly decreased INS levels（P<0.01）. Compared with the model group， the BP-1-50 group significantly reduced glucose levels and increased INS levels（P<0.05）. Histopathological examination of liver tissue revealed that various doses of BP-1 had a certain reparative effect on damaged liver tissue. The liver tissue structure in the BP-1-200 group was nearly normal， with hepatocytes appearing plump. Real-time PCR results showed that， compared with the blank group， the model group exhibited significantly upregulated mRNA expressions of Glucagon and PCK1， and significantly downregulated mRNA expression of Insa（P<0.01）. Compared with the model group， the BP-1-50 and BP-1-200 groups showed significantly downregulated mRNA expressions of Glucagon and PCK1， and significantly upregulated mRNA expression of Insa（P<0.01）. ConclusionThe semi-bionic extraction method for turnip polysaccharides yields a high extraction rate， is simple to operate， has low costs， making it suitable for large-scale industrial production. BP-1 consists of six monosaccharides， contains α-glycosidic bonds and uronic acids， exhibits hypoglycemic activity， and provides a certain protective effect on the liver of alloxan-induced diabetic model zebrafish.

ObjectiveTo optimize the extraction method for polysaccharides from turnip（Brassica rapa）， and analyze and evaluate the primary structure of the isolated and purified turnip polysaccharide fraction（BP-1） and its hypoglycemic effects in diabetic zebrafish. MethodsTaking polysaccharide yield as the evaluation index， a semi-bionic extraction method was employed. Single-factor experiments and Box-Behnken response surface methodology were used to investigate three factors of solid-to-liquid ratio， extraction time and extraction temperature， in order to optimize the extraction process. BP-1 was isolated and purified using the Sevage method and DEAE-52 cellulose column chromatography. Structural characterization of the turnip polysaccharides was performed using ultraviolet-visible spectrophotometry（UV）， gas chromatography-mass spectrometry（GC-MS）， Congo red assay， and Fourier-transform infrared spectroscopy（FT-IR） to determine purity， monosaccharide composition， triple-helix structure， and functional groups. The microstructure of the polysaccharides was observed using scanning electron microscopy（SEM） and atomic force microscopy（AFM）. Zebrafish were divided into the blank group（adding E3 medium）， and BP-1-1， BP-1-10， BP-1-50， BP-1-200， BP-1-1 000 groups（adding BP-1 solutions at concentrations of 1， 10， 50， 200， 1 000 mg·L^-1， respectively）， and zebrafish embryos were subjected to a 96-hour exposure experiment. The maximum tolerated concentration of BP-1 in zebrafish was determined by evaluating its effects on phenotype， survival rate， malformation rate， and heart rate. Experimental animals were randomly divided into the blank group， model group， BP-1-10 group（10 mg·L^-1）， BP-1-50 group（50 mg·L^-1）， and BP-1-200 group（200 mg·L^-1）. The blank group was cultured in E3 medium， the model and treatment groups were induced to establish a diabetic model in 4 day-post-fertilization（dpf） zebrafish embryos using 10 g·L^-1 of glucose combined with 500 µmol·L^-1 of alloxan. The treatment groups received corresponding doses of BP-1 solution， while the blank and model groups received an equal volume of saline. Glucose and insulin（INS） levels were measured using enzyme-linked immunosorbent assay（ELISA） kits， the effects on the liver were observed by hematoxylin-eosin（HE） histopathological sections. The mRNA expression levels of glucagon（Glucagon）， insulin（Insa）， and phosphoenolpyruvate carboxykinase 1（PCK1） were detected with real-time fluorescence quantitative polymerase chain reaction（Real-time PCR）. ResultsThe optimized extraction conditions were determined as follows：solid-to-liquid ratio of 1∶40（g·mL^-1）， extraction time of 66 min， and extraction temperature of 79 ℃. Under these conditions， the yield of turnip polysaccharides was （10.34±0.96）%. UV analysis indicated that BP-1 contained no proteins or nucleic acids， GC-MS analysis revealed that BP-1 consisted of six monosaccharides（arabinose， rhamnose， ribose， mannose， galactose and glucose）. Congo red assay indicated that the molecular conformation did not exhibit a triple-helix structure， FT-IR analysis showed the presence of α-glycosidic bonds and uronic acids， SEM analysis revealed an irregular flaky structure with a flat and smooth surface， AFM analysis suggested that the aggregated structure might be formed by the entanglement of molecular chains and intramolecular hydrogen bonding. The maximum tolerated concentration of BP-1 in zebrafish over 96 h was determined to be 200 mg·L^-1. Pharmacodynamic results showed that， compared with the blank group， the model group exhibited significantly increased glucose levels and significantly decreased INS levels（P<0.01）. Compared with the model group， the BP-1-50 group significantly reduced glucose levels and increased INS levels（P<0.05）. Histopathological examination of liver tissue revealed that various doses of BP-1 had a certain reparative effect on damaged liver tissue. The liver tissue structure in the BP-1-200 group was nearly normal， with hepatocytes appearing plump. Real-time PCR results showed that， compared with the blank group， the model group exhibited significantly upregulated mRNA expressions of Glucagon and PCK1， and significantly downregulated mRNA expression of Insa（P<0.01）. Compared with the model group， the BP-1-50 and BP-1-200 groups showed significantly downregulated mRNA expressions of Glucagon and PCK1， and significantly upregulated mRNA expression of Insa（P<0.01）. ConclusionThe semi-bionic extraction method for turnip polysaccharides yields a high extraction rate， is simple to operate， has low costs， making it suitable for large-scale industrial production. BP-1 consists of six monosaccharides， contains α-glycosidic bonds and uronic acids， exhibits hypoglycemic activity， and provides a certain protective effect on the liver of alloxan-induced diabetic model zebrafish.

ObjectiveTo investigate the effects of prostaglandin E₂ （PGE₂） microinjection into the median preoptic nucleus （MnPO） on core body temperature in female mice， and to clarify its underlying mechanism. MethodsMicroinjection cannula were implanted into the MnPO of female mice using stereotaxic surgery.Subsequently， a multi-channel temperature acquisition system was used to simultaneously monitor rectal and brown adipose tissue （BAT） temperatures before and after intra-MnPO injections of different reagents.To investigate the thermoregulatory effects of the microinjection of PGE₂ into the MnPO， 12 female C57BL/6 mice were randomly divided into a saline group （n=6） and a PGE₂ group （n=6）， which were injected with 0.1 μL saline and PGE₂ （2.8 mmol/L）， respectively.To determine whether E-series prostaglandin receptor （EP）1， EP3， and EP4 receptors mediate the thermoregulatory effects of PGE₂， 15 female C57BL/6 mice were randomly divided into 3 groups （n=5 per group）.Mice in each group first received an injection of 0.1 μL PGE2 （2.8 mmol/L） into the MnPO. After their body temperature returned to baseline levels， they were subsequently injected with a mixture of either EP1， EP3 or EP4 antagonist （ant） （20 mmol/L） + PGE2 （2.8 mmol/L）. ResultsCompared with baseline level， the rectal temperature （P<0.01） and BAT temperature （P<0.001） of female mice both increased significantly after microinjection of PGE₂ into the MnPO.Compared with the saline group， the increases in rectal temperature （P<0.001） and BAT temperature （P<0.000 1） were significantly greater in the PGE₂ group of mice.Furthermore， following the injection of PGE₂ into MnPO， the increase in BAT temperature was found to be significantly greater than that in rectal temperature in mice （P<0.001）.Compared to the administration of PGE₂ alone， co-injection of an EP3 ant + PGE₂ into the MnPO of mice resulted in a significantly smaller increase in both rectal temperature （P<0.001） and BAT temperature （P<0.001）.In contrast， the increases in rectal and BAT temperatures following MnPO injection of either EP1 ant + PGE₂ or EP4 ant + PGE₂ were not statistically significant （P>0.05）. ConclusionInjection of PGE₂ into the MnPO elevates BAT and core body temperature in female mice via the EP3 receptor.

Objective: To investigate the impact of RhE antigen-matched transfusion during liver transplantation on early postoperative recovery and complications. Methods: In this retrospective cohort study, ninety-five patients undergoing liver transplantation at Kunming First People's Hospital between January 2022 and July 2025 were enrolled. Patients were divided into two groups: Group 1 (RhE-mismatched transfusion, n=57) and Group 2 (RhE-matched transfusion, n=38). The baseline data, complete blood counts, hepatic and renal function, coagulation parameters, and complication rates between the two groups were compared at postoperative days 1, 3, 5, 7, and 10. Survival analysis was performed using the Kaplan-Meier method. Results: The baseline characteristics were well-balanced and comparable between the two groups (all P>0.05). The early postoperative mortality rate in the mismatched group (31.58%, 18/57) was significantly higher than that in the matched group (10.53%, 4/38) (P=0.017). The incidence of postoperative hepatic encephalopathy was significantly higher in the mismatched group (50.88%, 29/57) than in the matched group (10.53%, 4/38) (P<0.001). The incidence of postoperative haemorrhage in the mismatched group (24.56%, 14/57) was higher than that in the matched group (5.26%, 2/38), with a statistically significant difference (P=0.014). The incidence of perioperative infection in the mismatched group (28.07%, 16/57) was higher than that in the matched group (10.53%, 4/38), with a statistically significant difference (P=0.04). Corresponding odds ratios (OR) and 95% confidence intervals indicated a lower risk of these adverse events in the matched group. On postoperative day 1, the change in activated partial thromboplastin time (-1.6, 20.5) in the mismatched group was greater than in the matched group (-0.2, 5.5). The change in international normalised ratio (-0.56, 1.22) in the mismatched group was greater than in the matched group (-0.18, 0.32), while the change in albumin (-4.0, 4.8) was smaller in the mismatched group than in the matched group (-2.5, 8.8). On postoperative day 5, the change in albumin (-0.41±7.83) in the mismatched group was smaller than in the matched group (2.68±4.53). At postoperative day 7, the change in albumin in the mismatched group (-0.61±7.38) was smaller than that in the matched group (2.51±5.85), while the change in D-dimer in the mismatched group (0.73, 7.4) was greater than that in the matched group (-1.6, 4.3). On postoperative day 10, the mismatched group exhibited significantly higher fibrinogen levels (-1.21, 1.78) than the matched group (-0.49, 0.97), and significantly longer prothrombin times (-11.3, -2.7) than the matched group (-6.2, -0.8) (all P<0.05). The matched group exhibited a mean overall survival (OS) of 32.803 months (95% CI:29.171-36.436 months), significantly exceeding the mismatched group's 28.996 months (95% CI:24.202-33.790 months). The log-rank test yielded statistically significant results (χ =4.307, P=0.038). Conclusion: Implementing RhE blood group-matched transfusion during liver transplantation may help reduce early postoperative mortality and the incidence of major complication rates, promote faster recovery of coagulation and liver function, and thereby improve short-term patient outcomes.

ObjectiveTo investigate the therapeutic effect of Astragali Radix-mediated changes in gut microbiota on treating type 2 diabetes （T2DM）. MethodsA 12-week randomized， placebo-controlled clinical trial enrolled eighty patients with newly diagnosed type 2 diabetes and poor glycemic control in the Qi deficiency type. All patients received insulin therapy. The observation group （40 cases） was administered with Astragali Radix Granules， while the control group （40 cases） received a placebo. Both treamtents were taken orally twice daily. Changes in gut microbiota were assessed by 16s rDNA sequencing. Serum glucagon-like peptide-1 （GLP-1） levels were measured using enzyme-linked immunosorbent assay （ELISA）. Glucose metabolism indicators including fasting blood glucose （FPG）， 2-hour postprandial blood glucose （2 h PG），glycated albumin（GA）， and glycated hemoglobin （HbA1c） were evaluated. Pancreatic function was evaluated using fasting C-peptide （FCP）， 2-hour postprandial C-peptide （2 h CP）， and C-peptide area under the curve （AUC_cp）. Traditional Chinese medicine （TCM） syndrome scores， clinical efficacy， and safety indicators were also observed. ResultsIn terms of glucose metabolism indicators， compared with the baseline， both groups exhibited significantly lower FPG， 2 h PG， GA and HbA1C （P<0.01），while FCP， 2 h CP and AUC_cp were significantly higher （P<0.01）. Compared with the control group after the treatment， the observation group showed significantly lower FPG， 2 h PG， GA and HbA1C（P<0.05， P<0.01），and significantly higher FCP， 2 h CP and AUC_cp （P<0.05， P<0.01）， indicating that Astragali Radix can improve glucose metabolism. In terms of the diversity of gut microbiota， no significant differences were detected in the Chao1， Shannon and Simpson indexes of the two groups compared with their respective baselines. However， compared with the post-treatment control group， the observation group demonstrated significant increases in the Chao1， Shannon and Simpson indexes （P<0.05， P<0.01）. The β-diversity analysis showed significant separation in gut microbiota composition before and after treatment in both groups， indicating that Astragali Radix can significantly alter the structure and improve the diversity of gut microbiota. At the phylum level， compared with the baseline， both groups showed a significant increase in the relative abundance of Bacteroidota（P<0.01）. The relative abundance of the potentially harmful phylum Proteobacteria was significantly lower in the observation Group after treatment （P<0.01）. Compared with the post-treatment control group， the observation group had a significantly higher relative abundance of Bacteroidota（P<0.01）. No significant difference was found in Firmicutes/Bacteroidota （F/B） ratio between the two groups after treatment， and other phyla showed no significant differences. At the genus level， compared with the baseline， the observation group exhibited a significant increase in Bacteroides （P<0.01） and a significant decrease in Escherichia-Shigella （P<0.01）， whereas no significant difference was seen in the control group . Compared with the control group after treatment， the observation group after treatment had a significantly higher relative abundance of Bacteroides （P<0.01）. No significant differences were seen in other genera. Linear discriminant analysis （LDA） identified potential characteristics taxa： in the observation group， Bacteroidota at the phylum level and Bacteroides and Dubosiella at the genus level， in the control group， Proteobacteria at the phylum level as well as Barnesiella and Staphylococcus at the genus level. Correlation analysis based on a heatmap revealed that GLP-1 levels were positively correlated with Firmicutes， F/B ratio and Fusobacterium， and negatively correlated with Bacteroidota， Proteobacteria， Bacteroides and Escherichia-Shigella. In terms of clinical efficacy， compared with the control group， the total effective rate of the observation group was significantly higher （P<0.05）. Compared with the baseline， the scores for shortness of breath， fatigue， weakness， spontaneous sweating and reluctance to speak significantly decreased in both groups （P<0.01）. Compared with the control group after treatment， the score for weakness was significantly lower in the observation group （P<0.01），indicating that Astragali Radix could improve clinical symptoms and alleviate weakness symptoms. In terms of safety， compared with the baseline， alanine aminotransferase （ALT） levels significantly decreased in both groups （P<0.05，P<0.01），indicating that Astragali Radix did not induce any significant abnormalities in liver and kidney functions. ConclusionAstragali Radix demonstrates the potential to significantly improve the gut microbiota environment in patients of newly diagnosed type 2 diabetes with Qi deficiency. The therapeutic effect may contribute to glycemic control， possibly mediated by an elevation in GLP-1 level. These findings may support its further clinical investigations and potential applications.

ObjectiveMitochondria are not only the central organelles responsible for cellular energy metabolism but also play essential roles in regulating cell cycle progression and cytoskeletal dynamics. In recent years, accumulating evidence has demonstrated that mitochondrial homeostasis is closely associated with mitotic progression and cytokinesis. Schizosaccharomyces pombe serves as a classical and well-established model organism. Because its cell cycle regulatory mechanisms are highly conserved throughout evolution, its genetic background is clearly defined, and experimental manipulation is efficient and convenient, it has been extensively applied in studies of cell growth, division, and reproductive mechanisms. The SPBC1604.04 gene encodes a previously uncharacterized mitochondrial carrier protein in Schizosaccharomyces pombe. This gene is located on chromosome II and spans 1 018 base pairs in length. It encodes a protein consisting of 238 amino acids with a predicted molecular mass of approximately 31.03 ku. Bioinformatic analysis predicts that this protein is responsible for the transport of thiamine pyrophosphate (TPP) into mitochondria. However, the effects of SPBC1604.04 gene deletion on mitotic cell dynamics under different temperature conditions have not been fully elucidated. MethodsThe SPBC1604.04 deletion strain of Schizosaccharomyces pombe was used as the experimental model. Fluorescent protein markers were constructed in the deletion background to label mitochondria, microtubules, actin, myosin, the nuclear envelope, and chromosomes. Live-cell imaging was performed using a TCS-SP8 laser scanning confocal microscope under normal temperature conditions (25℃) and heat stress conditions (37℃). Time-lapse microscopy was applied to dynamically monitor mitochondrial morphology and distribution, spindle assembly and elongation, chromosome segregation, as well as the formation and constriction of the actomyosin ring during cytokinesis. ImageJ software was used for quantitative measurements, including microtubule length during mitosis, spindle length at different mitotic stages, mitochondrial fluorescence intensity as an indicator of mitochondrial content, actomyosin ring length, nuclear envelope area, and chromosome segregation timing. Statistical analyses were conducted to compare phenotypic differences between the wild-type and SPBC1604.04 deletion strains at both temperature conditions. Through these analyses, we systematically investigated the impact of SPBC1604.04 deletion on mitotic cell dynamics in fission yeast under both normal physiological conditions and temperature stress. ResultsAt 25℃, compared with wild-type cells, the SPBC1604.04Δ strain exhibited a pronounced tendency toward mitochondrial fragmentation, accompanied by abnormal mitochondrial content and a significant reduction in mitochondrial fluorescence intensity. These observations suggest impaired mitochondrial homeostasis under normal growth conditions. In addition, the constriction time of actomyosin ring during cytokinesis was markedly prolonged, indicating that deletion of SPBC1604.04 affects the dynamics of the contractile machinery. However, no obvious defects were observed in spindle assembly, spindle elongation, or chromosome segregation. Under heat stress at 37℃, mitochondrial morphology in the SPBC1604.04Δ strain showed a tendency to recover toward a continuous tubular network structure. Mitochondrial content was restored, fluorescence intensity increased, and the constriction time of the actomyosin ring returned to levels comparable to those of wild-type cells. These results indicate that the mitotic defects observed at normal temperature are partially or fully alleviated under heat stress conditions. ConclusionThis study demonstrates that deletion of the SPBC1604.04 gene leads to abnormal mitochondrial content in Schizosaccharomyces pombe. The mitochondrial carrier protein SPBC1604.04 participates in regulating actomyosin ring constriction during mitosis but does not appear to be directly involved in the regulation of spindle dynamics or chromosome segregation. Our findings provide key experimental evidence for understanding the functional link between the SPBC1604.04 gene, mitochondrial homeostasis, and mitotic regulation.

ObjectiveMitochondria are not only the central organelles responsible for cellular energy metabolism but also play essential roles in regulating cell cycle progression and cytoskeletal dynamics. In recent years, accumulating evidence has demonstrated that mitochondrial homeostasis is closely associated with mitotic progression and cytokinesis. Schizosaccharomyces pombe serves as a classical and well-established model organism. Because its cell cycle regulatory mechanisms are highly conserved throughout evolution, its genetic background is clearly defined, and experimental manipulation is efficient and convenient, it has been extensively applied in studies of cell growth, division, and reproductive mechanisms. The SPBC1604.04 gene encodes a previously uncharacterized mitochondrial carrier protein in Schizosaccharomyces pombe. This gene is located on chromosome II and spans 1 018 base pairs in length. It encodes a protein consisting of 238 amino acids with a predicted molecular mass of approximately 31.03 ku. Bioinformatic analysis predicts that this protein is responsible for the transport of thiamine pyrophosphate (TPP) into mitochondria. However, the effects of SPBC1604.04 gene deletion on mitotic cell dynamics under different temperature conditions have not been fully elucidated. MethodsThe SPBC1604.04 deletion strain of Schizosaccharomyces pombe was used as the experimental model. Fluorescent protein markers were constructed in the deletion background to label mitochondria, microtubules, actin, myosin, the nuclear envelope, and chromosomes. Live-cell imaging was performed using a TCS-SP8 laser scanning confocal microscope under normal temperature conditions (25℃) and heat stress conditions (37℃). Time-lapse microscopy was applied to dynamically monitor mitochondrial morphology and distribution, spindle assembly and elongation, chromosome segregation, as well as the formation and constriction of the actomyosin ring during cytokinesis. ImageJ software was used for quantitative measurements, including microtubule length during mitosis, spindle length at different mitotic stages, mitochondrial fluorescence intensity as an indicator of mitochondrial content, actomyosin ring length, nuclear envelope area, and chromosome segregation timing. Statistical analyses were conducted to compare phenotypic differences between the wild-type and SPBC1604.04 deletion strains at both temperature conditions. Through these analyses, we systematically investigated the impact of SPBC1604.04 deletion on mitotic cell dynamics in fission yeast under both normal physiological conditions and temperature stress. ResultsAt 25℃, compared with wild-type cells, the SPBC1604.04Δ strain exhibited a pronounced tendency toward mitochondrial fragmentation, accompanied by abnormal mitochondrial content and a significant reduction in mitochondrial fluorescence intensity. These observations suggest impaired mitochondrial homeostasis under normal growth conditions. In addition, the constriction time of actomyosin ring during cytokinesis was markedly prolonged, indicating that deletion of SPBC1604.04 affects the dynamics of the contractile machinery. However, no obvious defects were observed in spindle assembly, spindle elongation, or chromosome segregation. Under heat stress at 37℃, mitochondrial morphology in the SPBC1604.04Δ strain showed a tendency to recover toward a continuous tubular network structure. Mitochondrial content was restored, fluorescence intensity increased, and the constriction time of the actomyosin ring returned to levels comparable to those of wild-type cells. These results indicate that the mitotic defects observed at normal temperature are partially or fully alleviated under heat stress conditions. ConclusionThis study demonstrates that deletion of the SPBC1604.04 gene leads to abnormal mitochondrial content in Schizosaccharomyces pombe. The mitochondrial carrier protein SPBC1604.04 participates in regulating actomyosin ring constriction during mitosis but does not appear to be directly involved in the regulation of spindle dynamics or chromosome segregation. Our findings provide key experimental evidence for understanding the functional link between the SPBC1604.04 gene, mitochondrial homeostasis, and mitotic regulation.

Background/Aims: Dysregulated cholesterol metabolism is a hallmark of hepatocellular carcinoma (HCC) that drives tumor initiation and progression. However, clinical targeting of cholesterol metabolism has yielded limited benefits due to stringent feedback in tumor cells. Identifying a central mediator capable of restoring cholesterol homeostasis within the cell’s intrinsically fine-tuned regulatory framework is urgently needed. Methods: We integrated a proteomic dataset from patients with cholesterol-dysregulated HCC into a global cholesterol metabolic regulatory network to identify potential therapeutic targets for disrupted cholesterol homeostasis. The prognostic significance of the candidate targets was further validated in an independent cohort through immunohistochemistry. Functional and mechanistic studies were conducted in vitro using HCC cell lines and in vivo using mouse models. The pharmacological efficacy of the candidate agent was evaluated in both subcutaneous and orthotopic HCC mouse models. Results: ER lipid raft-associated 1 (ERLIN1), a pivotal regulator of cholesterol metabolism reprogramming, was identified as an independent favorable prognostic indicator in HCC. ERLIN1 constrains HCC progression both in vitro and in vivo by stabilizing the INSIG1–SCAP–SREBP2 axis and maintaining the metabolic balance of intracellular cholesterol. Under hypoxia, impaired factor-inhibiting hypoxia-1-dependent hydroxylation of ASB11 at asparagine residues 90 and 92 enhances ASB11-mediated ERLIN1 degradation. Pharmacological targeting of this axis using zoledronic acid (ZoA) attenuated HCC progression by weakening the ASB11–ERLIN1 interaction and restoring cholesterol homeostasis. Conclusions ERLIN1 represents a druggable metabolic vulnerability in cholesterol-dysregulated HCC. Targeting the ASB11–ERLIN1 axis with the clinically approved ZoA reestablishes cholesterol homeostasis and offers a promising therapeutic strategy to overcome the current limitations of cholesterol-targeted HCC therapies.

OBJECTIVE: To standardize the operative procedure for tissue-engineered cartilage repair, by demonstrating surgical technique of arthroscopic implantation of decalcified cortex-cancellous bone scaffolds, and summarizing the surgical experience of the sports medicine department team at Peking University Third Hospital. METHODS: This article elaborates on surgical techniques and skills, focusing on the unabridged implantation technology and surgical procedure of decalcified cortex-cancellous bone scaffolds under arthroscopy: First, the patient was placed in the supine position. After anesthesia had been established, the surgeon established an arthroscope and explored the damaged area under the scope. After confirming the size and location of the injury site, the surgeon cleaned the damaged cartilage, and also trimmed the edges of the cartilage to ensure that the cut surface was smooth and stable. the surgeon performed the micro-fracture surgery in the area of cartilage injury, and then measured the size of the injured area under the scope. Next, the surgeon manually trimmed the tissue-engineered scaffold based on the measurements taken under the arthroscope, and then directly implanted the scaffold using a sleeve. A honeycomb-shaped fixator was used to implant absorbable nails to fix the scaffold. After the scaffold was installed, the knee was repeatedly flexed and extended for 10-20 times to ensure stability and range of motion. Finally, the arthroscope was withdrawn and the wound was closed. RESULTS: Decalcified cortex-cancellous bone scaffolds possessed unparalleled advantages over synthetic materials in terms of morphology and biomechanics. The cancellous bone part of the scaffold provided a three-dimensional, porous space for cell growth, while the cortical bone part offered the necessary mechanical strength. The surgery was performed entirely under arthroscopy to minimize invasiveness to the patient. Absorbable pins were used for fixation to ensure the stability of the scaffold. This technique could effectively improve the prognosis of the patients with cartilage injuries and standardized the surgical procedures for arthroscopic tissue-engineered scaffold operations in the patients with cartilage damage. CONCLUSION With the standard arthroscopic tissue-engineered scaffold repair technique, it is possible to successfully repair damaged cartilage, alleviate symptoms in the short term, and provide a more ideal long-term prognosis. The author and their team explain the surgical procedures for tissue-engineered scaffolds under arthroscopy, with the aim of guiding future clinical practice.
Tissue Engineering/methods* ; Humans ; Tissue Scaffolds ; Arthroscopy/methods* ; Cartilage, Articular/surgery*

BACKGROUND: Biomarkers-based prediction of long-term risk of acute coronary syndrome (ACS) is scarce. We aim to develop a risk score integrating clinical routine information (C) and plasma biomarkers (B) for predicting long-term risk of ACS patients. METHODS: We included 2729 ACS patients from the OCEA (Observation of cardiovascular events in ACS patients). The earlier admitted 1910 patients were enrolled as development cohort; and the subsequently admitted 819 subjects were treated as validation cohort. We investigated 10-year risk of cardiovascular (CV) death, myocardial infarction (MI) and all cause death in these patients. Potential variables contributing to risk of clinical events were assessed using Cox regression models and a score was derived using main part of these variables. RESULTS: During 16,110 person-years of follow-up, there were 238 CV death/MI in the development cohort. The 7 most important predictors including in the final model were NT-proBNP, D-dimer, GDF-15, peripheral artery disease (PAD), Fibrinogen, ST-segment elevated MI (STEMI), left ventricular ejection fraction (LVEF), termed as CB-ACS score. C-index of the score for predication of cardiovascular events was 0.79 (95% CI: 0.76-0.82) in development cohort and 0.77 (95% CI: 0.76-0.78) in the validation cohort (5832 person-years of follow-up), which outperformed GRACE 2.0 and ABC-ACS risk score. The CB-ACS score was also well calibrated in development and validation cohort (Greenwood-Nam-D'Agostino: P = 0.70 and P = 0.07, respectively). CONCLUSIONS CB-ACS risk score provides a useful tool for long-term prediction of CV events in patients with ACS. This model outperforms GRACE 2.0 and ABC-ACS ischemic risk score.