Chemotherapy is currently the mainstay of systemic management for triple-negative breast cancer (TNBC), but chemoresistance significantly impacts patient outcomes. Our research indicates that Doxorubicin (Dox)-resistant TNBC cells exhibit increased glycolysis and ATP generation compared to their parental cells, with this metabolic shift contributing to chemoresistance. We discovered that ALKBH3, an m¹A demethylase enzyme, is crucial in regulating the enhanced glycolysis in Dox-resistant TNBC cells. Knocking down ALKBH3 reduced ATP generation, glucose consumption, and lactate production, implicating its involvement in mediating glycolysis. Further investigation revealed that aldolase A (ALDOA), a key enzyme in glycolysis, is a downstream target of ALKBH3. ALKBH3 regulates ALDOA mRNA stability through m¹A demethylation at the 3'-untranslated region (3'UTR). This methylation negatively affects ALDOA mRNA stability by recruiting the YTHDF2/PAN2-PAN3 complex, leading to mRNA degradation. The ALKBH3/ALDOA axis promotes Dox resistance both in vitro and in vivo. Clinical analysis demonstrated that ALKBH3 and ALDOA are upregulated in breast cancer tissues, and higher expression of these proteins is associated with reduced overall survival in TNBC patients. Our study highlights the role of the ALKBH3/ALDOA axis in contributing to Dox resistance in TNBC cells through regulation of ALDOA mRNA stability and glycolysis.

The cardioprotective effects of histone deacetylase (HDAC) inhibitors (HDIs) are at odds with the deleterious effects of HDAC depletion. Here, we use HDAC3 as a prototype HDAC to address this contradiction. We show that adult-onset cardiac-specific depletion of HDAC3 in mice causes cardiac hypertrophy and contractile dysfunction on a high-fat diet (HFD), excluding developmental disruption as a major reason for the contradiction. Genetically abolishing HDAC3 enzymatic activity without affecting its protein level does not cause cardiac dysfunction on HFD. HDAC3 depletion causes robust downregulation of lipid oxidation/bioenergetic genes and upregulation of antioxidant/anti-apoptotic genes. In contrast, HDAC3 enzyme activity abolishment causes much milder changes in far fewer genes. The abnormal gene expression is cardiomyocyte-autonomous and can be rescued by an enzyme-dead HDAC3 mutant but not by an HDAC3 mutant (Δ33-70) that lacks interaction with the nuclear-envelope protein lamina-associated polypeptide 2β (LAP2β). Tethering LAP2β to the HDAC3 Δ33-70 mutant restored its ability to rescue gene expression. Finally, HDAC3 depletion, not loss of HDAC3 enzymatic activity, exacerbates cardiac contractile functions upon aortic constriction. These results suggest that the cardiac function of HDAC3 in adults is not attributable to its enzyme activity, which has implications for understanding the cardioprotective effects of HDIs.

Objective:To explore the characteristics of 24-hour movement in preschoolers and its relationship with executive function.Methods:The study subjects consisted of 555 preschoolers in Zhuhai City. An objective measurement approach was employed, physical activity level was assessed by utilizing accelerometers and executive function was evaluated by computerized behavioral tests.The characteristics of 24-hour movement in children were described.Data were analyzed using SPSS 26.0 software, independent samples t-test was used to compare differences in 24-hour activity behaviors across genders, and multivariate linear regression was used to explore the effect of preschoolers' achievement of activity behaviors on executive functioning. Results:According to the physical activity guideline for Chinese preschoolers aged 3-6 years recommendations for children's daily moderate to vigorous-intensity physical activity, sedentary screen time, and sleep duration, among the study participants, the compliance rate of moderate to vigorous-intensity physical activity (≥60 min/d) was 33.9 %(188/555), sedentary screen time (≤1 h/d) was 43.4%(241/555), and sleep duration (10-13 h/d) was 83.4 %(463/555). Only 13.3 %(74/555) of the participants met the recommended guidelines for all three behaviors. The correct score of the working memory test was (60.6±20.3) and the reaction time of the inhibitory control test was (695.1±135.8) ms. In male preschoolers, moderate to vigorous-intensity physical activity (≥60 min/d) positively predicted working memory( β=0.121, P=0.037), while sedentary screen time (≤1 h/d) negatively predicted inhibitory control ( β=-0.122, P=0.036). Conclusions:The daily compliance rate of moderate to vigorous-intensity physical activity and sedentary screen time is low among preschooler. The former is positively correlated with working memory in preschool boys, while the latter is negatively correlated with inhibitory control.

The NMGCF-19 strain is an E.coli strain isolated and identified in our laboratory from lambs manifesting severe diarrhea and meningitis.Previous analysis of the genome sequence of NMGCF-19 strain showed that the outer membrane protein A(ompA)gene was a potential viru-lent gene.In order to determine whether the ompA gene is associated with the pathogenicity of NMGCF-19 strain and the underlying mechanism,the NMGCF-19 strain with ompA knockout(NMGCF-19△ompA)was generated in this study using CRISPR/Cas9 technology and used to de-termine the role of ompA gene in mediating the encephalitis by NMGCF-19 infection and the un-derlying mechanism using the mouse model system.The results showed that the neuronal cell nec-rosis in the hippocampus in mice infected by NMGCF-19△ompA was significantly reduced and was not focal compared with that of mice infected with the wild-type NMGCF-19 strain.The number of bacteria in brain of mice infected by NMGCF-19 △ompA was significantly reduced in comparison to that of mice infected by NMGCF-19.Simultaneously,the mRNA and protein expression levels of the tight junction proteins ZO-1 and Occludin were both increased in mice infected by NMGCF-19 △ompA strain compared with the mice infected by NMGCF-19 strain.These results suggest that the ompA gene is a virulent gene and plays an important role in the invasion of the blood-brain barrier by NMGCF-19 strain in mice.

AIM:To investigate the effect of doublecortin-like kinase 1(DCLK1)on the biological properties of gastric cancer stem cells,and to explore its possible mechanism.METHODS:Serum-free suspension culture of gastric cancer stem cells and targeted inhibition of DCLK1 activity in gastric cancer stem cells with DCLK1 inhibitor DCLK1-IN-1 were performed.The expression levels of DCLK1,stemness-related proteins(SOX2 and OCT4),proliferation-related pro-teins(cyclin D1 and c-MYC),drug resistance-related proteins(ABCG2 and TOP2A),epithelial-mesenchymal transition-related proteins(E-cadherin,vimentin and Snail),and PI3K/AKT/mTOR signaling pathway-related proteins in gastric cancer stem cells were examined by Western blot.The effects of DCLK1 on viability and drug resistance of gastric cancer stem cells were determined by CCK-8 assay,and the effects of DCLK1 on self-renewal of gastric cancer stem cells were de-termined by methylcellulose spheroid-forming assay.Wound-healing and Transwell assays were performed to assess the ef-fect of DCLK1 on the migration and invasion of gastric cancer stem cells.RESULTS:The expression levels of DCLK1 and stemness-related proteins SOX2 and OCT4 in gastric cancer stem cells were significantly higher than those in parental cells(P＜0.01).The proliferation,drug resistance,migration and invasion of gastric cancer stem cells in DCLK1 inhibi-tion group were significantly lower than those in Sphere cell group(P＜0.01).The expression levels of proliferation-related proteins(c-MYC and cyclin D1)and drug resistance-related proteins(TOP2A and ABCG2)were down-regulated,the ex-pression of epithelial marker E-cadherin was up-regulated,the expression of mesenchymal markers vimentin and Snail was down-regulated,and the expression levels of PI3K/AKT/mTOR signaling pathway-related proteins and their phosphoryla-tion levels were reduced in DCLK1 inhibition group(P＜0.05).CONCLUSION:DCLK1 is highly expressed in gastric cancer stem cells,which may be involved in the proliferation,drug resistance and invasion of gastric cancer stem cells by regulating PI3K/AKT/mTOR signaling pathway.It suggests that DCLK1 can be used as a potential target for gastric cancer stem cells.

OBJECTIVE: To summarize the research progress of bio-derived materials used for bladder regeneration and repair. METHODS: The recent domestic and foreign sutudies on bio-derived materials used for bladder regeneration and repair, including classification, morphology optimization process, tissue regeneration strategies, and relevant clinical trials, were summarized and analyzed. RESULTS: Numerous types of bio-derived materials are employed in bladder regeneration and repair, characterized by their low immunogenicity and high inducible activity. Surface modification, gelation, and other morphology optimization process have significantly broadened the application scope of bio-derived materials. These advancements have effectively addressed complications, such as perforation and urolith formation, that may arise during bladder regeneration and repair. The strategy of tissue regeneration utilizing bio-derived materials, targeting the regeneration of bladder epithelium, smooth muscle, blood vessels, and nerves, offers a novel approach to achieving functional regeneration of bladder. Bio-derived materials show great promise for use in bladder regeneration and repair, yet the results from clinical trials with these materials have been less than satisfactory. CONCLUSION Bio-derived materials are widely used in bladder regeneration and repair due to the good biocompatibility, low immunogenicity, and degradable properties, yet face a series of problems, and there are no commercialized bladder tissue engineering grafts used in clinical treatment.
Humans ; Tissue Engineering/methods* ; Urinary Bladder/surgery* ; Biocompatible Materials/chemistry* ; Regeneration ; Tissue Scaffolds ; Animals

With the advances in understanding the relationships among biomaterials, the immune system and the skeletal system, the host responses elicited by implanted biomaterials can be balanced by properly designing material characteristics from the perspective of osteoimmunology. The immunoregulatory properties of bone tissue engineering scaffolds provide advantages for inducing macrophages from the proinflammatory M1 phenotype to the anti-inflammatory M2 phenotype and promoting osseointegration. Hydrogels are increasingly a focus in bone tissue engineering, and the immune response can be affected by different compositions of hydrogels, such as the sources, concentration, molecular weight, coupling with fibronectin, and the addition of cross-linking agents. Different physicochemical properties of modified hydrogels can trigger different host immune responses, modified by using soft photolithography to fabricate micropatterned hydrogels, adding enzyme-sensitive sequences, ester bonds and dynamic covalent chemistry to prevent rapid or slow degradation of the hydrogels, and adding porogens and 3D printing to modify the hydrogels with macroporous interconnective pore structures, soft and injectable hydrogels, etc. These optimized hydrogels can reduce proinflammatory factors, promote M2 macrophage polarization, and minimize foreign body reactions to evoke bone regeneration. However, the mechanism underlying the bone immune response is still poorly understood, and further study of the effects of hydrogels with different physicochemical properties on immune regulation is needed.

Qinghai University lies in western China. There are some gaps between western and eastern universities at student level, education fund, experiment condition and technique supply for graduate students. To improve educational quality for graduate students, Department of Pathophysiology of Qinghai University focused on teaching methods, evaluation approaches and capacity of teachers and constructed a course named Clinical Pathophysiology for scientific research graduate students. The course adopted a series of measures such as combining lectures with students' self-learning (PBL, paper discussion, lecture for students), introducing experts from other schools to give lectures, establishing formative evaluation, optimizing teacher's group and so on, which have gained some achievements and improved the training quality for graduate students.

Objective: To investigate the role of peroxisome proliferator-activated-receptor-γ coactivator-1α (PGC-1α) in carotid atherosclerosis due to human cytomegalovirus (HCMV) infection. Methods: Thirty-three samples of carotid arterial sclerosis plaques after carotid endarterectomy (CEA) were collected in the experiment group, and 12 pieces of normal intracranial arteries were collected in the control group. The plaques of carotid artery were studied using in situ hybridization (ISH) and immunohistochemistry (IHC) for the role of PGC-1α in HCMV related atherosclerosis. HCMV immediate early (IE) gene of two groups was detected using ISH, and the expression of PGC-1α using IHC. Results: HCMV IE gene was positive 72.7% in the atherosclerotic plague group, while 16.6% in the control group, there was significant difference between the two groups (P<0.05). As to PGC-1α expression, in the atherosclerosis group, 8/33(24.2%) were strongly positive (+ + + ), 9/33(27.2%) were positive (+ + ), 6/33(18.2%) were weakly positive (+ ), and 10/33(30.3%) were negative (-). While in the control group, 1/12(8.3%) was strongly positive (+ + + ), 2/12(16.7%) were positive (+ + ), and 1/12 (8.3%) was weakly positive (+ ), and 8/12 (66.7%) were negative (-). The differences between the two groups were statistically significant (P<0.05). Conclusions HCMV immediate early gene and PGC-1α were both related to athersclerotic plaque formation, suggesting that PGC-1α may play an important role in as the plaque formation due to HCMV infection.

Objective: To investigate the role of lectin-like Ox-LDL receptor-1( LOX-1) in the activation and oxidative stress of cultured human umbilical vein endothelial cell(HUVEC) after human cytomegalovirus(HCMV) infection. Methods: HUVEC were divided into four groups: HCMV, Control, Carrageenan, and HCMV+ Carrageenan. After HCMV AD169 infection, the supernatant of the culture was extracted, and cells were lysed. The levels of LOX-1 mRNA, intercellular adhesion molecule-1(ICAM-1) mRNA and vascular cellular adhesion molecule-1(VCAM-1) in HUVEC were measured by real-time PCR. And the content of nitrogen monoxidum(NO) of the supernatant was detected by nitrate reductase method accordingly. Results: 24 h after infection, the mRNA expression of LOX-1, ICAM-1 and VCAM-1 in HUVEC of HCMV infected group increased obviously compared to control, and NO quantity increased accordingly. The mRNA expression of LOX-1, ICAM-1 and VCAM-1 and the quantity of NO decreased after adding the LOX-1 inhibitor carrageenan. There was significant difference between groups(P<0.05). Conclusions HCMV may increase the mRNA expression of ICAM-1 and VCAM-1 and quantity of NO by upregulating the mRNA expresion of LOX-1, which may contribute to the formation of a therosclerosis(AS).