Objective: To assess the association between the metabolic score for insulin resistance index (METS-IR) and overactive bladder (OAB) in the US population，so as to explore the potential of METS-IR as a predictive tool for OAB risk and to provide insights for early screening and intervention strategies. Methods: Based on the data from the National Health and Nutrition Examination Survey (NHANES) 2005-2018，a cross-sectional design was employed，and multivariate logistic regression models were used to analyze the association between METS-IR and OAB. METS-IR was analyzed both as a continuous variable and categorized into quartiles. To further validate the association between METS-IR and OAB across diverse populations，subgroup analyses were conducted in participants stratified by clinical characteristics. Smooth curve fitting was employed to test the linearity of the METS-IR-OAB relationship. Results: Elevated METS-IR was associated with an increased risk of OAB (P<0.001)，and this positive correlation remained stable when METS-IR was categorized into quartiles (P<0.001). Subgroup analyses revealed that the association between METS-IR and OAB was more pronounced in females，participants younger than 55 years，and non-diabetic individuals (P<0.05). Furthermore，smooth curve fitting confirmed a linear positive correlation between METS-IR and OAB，with this linear relationship observed in both diabetic and non-diabetic groups. Conclusion: This study，based on the NHANES 2005-2018 database，found a linear positive correlation between METS-IR and OAB.

BACKGROUND:Exercise training can improve osteoporosis,but its effects and mechanisms on senile osteoporosis are not fully understood. OBJECTIVE:To observe the effect of treadmill exercise on osteoporosis and wnt/β-catenin signal pathway in aged rats. METHODS:Sixteen 24-month-old male Sprague-Dawley rats were randomly divided into osteoporosis group(n=8)and treadmill group(n=8)and eight 6-month-old male Sprague-Dawley rats were used as young control group.The model of senile osteoporosis was replicated by natural aging and the rats in the treadmill group were treated with treadmill exercise once a day,5 days a week,for 8 weeks.Levels of bone metabolic markers such as type I collagen cross-linked C-terminal peptide,tartrate resistant acid phosphatase,osteocalcin and bone specific alkaline phosphatase were detected by ELISA;bone mineral density of the left femur and L5 was measured by dual energy X-ray;bone scanning and bone microstructure quantitative analysis were performed by bone micro-CT;and the mRNA and protein expression levels of wnt3a,β-catenin,LRP5,DKK1 and GSK3β were detected by RT-PCR and western blot,respectively. RESULTS AND CONCLUSION:Compared with the young control group,the osteoporosis group showed a reduction in serum bone specific alkaline phosphatase and osteocalcin levels(P<0.05),bone mineral density of the femur and L5,the number of tibia and L4 bone trabeculae,bone volume,bone volume fraction(P<0.05),and mRNA and protein expression of wnt3a,β-catenin,and LRP5 in bone marrow tissue(P<0.05)as well as an increase in serum levels of tartrate resistant acid phosphatase and type I collagen cross-linked C-terminal peptide(P<0.05),the intertrabecular space between the tibia and L4,structural model index(P<0.05),and mRNA and protein expression of DKK1 and GSK3 β in bone marrow tissue(P<0.05).In addition to the reduced number of trabeculae in the tibia and L4 vertebrae,the trabeculae were structurally disturbed and sparsely aligned and fractured.Compared with the osteoporosis group,the treadmill group showed an increase in serum bone specific alkaline phosphatase and osteocalcin levels(P<0.05),bone mineral density of the femur and L5(P<0.05),the number of tibial trabeculae,bone volume,bone volume fraction(P<0.05),mRNA and protein expression of wnt3a,β-catenin,and LRP5 in bone marrow tissue(P<0.05)but a reduction in the serum levels of tartrate resistant acid phosphatase and type I collagen cross-linked C-terminal peptide,L4 trabecular space,tibial trabecular space,structural model index,and mRNA and protein expression of DKK1 and GSK3 β in bone marrow tissue(P<0.05).In addition to the increased number of tibial and L4 trabeculae,the trabeculae were arranged in a regular and dense pattern and were connected to a network.To conclude,treadmill exercise may improve osteoporosis in aged rats by activating the wnt/β-catenin signal pathway.

Objective To investigate the effect of type 2 dengue virus(DENV-2)infection on autophagy in human umbilical vein endothelial cells(HUVECs)and the mechanism mediating the inhibitory effect of baicalin against DENV-2 infection.Methods Cultured HUVECs with DENV-2 infection were treated with different concentrations of baicalin,and the changes in autophagy of the cells were detected using transmission electron microscopy.Lyso Tracker Red staining was used to examine pH changes in the lysosomes of the cells,and the expressions of ATG5,beclin-1,LC3,P62,STX17,SNAP29,VAMP8,and PI3K/AKT signaling pathway-related proteins were detected by Western blotting.DENV-2 replication in the cells were evaluated using RT-qPCR.The differentially expressed proteins in DENV-2-infected HUVECs were identified by proteomics screening.Results Treatment with baicalin did not significantly affect the viability of cultured HUVECs.Proteomic studies suggested that the PI3K-AKT pathway played an important role in mediating cell injury induced by DENV-2 infection.The results of RT-qPCR demonstrated that baicalin dose-dependently inhibited DENV-2 replication in HUVECs and produced the strongest inhibitory effect at the concentration of 50 μg/mL.Transmission electron microscopy,Lyso Tracker Red staining,RT-qPCR,and Western blotting all showed significant inhibitory effect of baicalin on DENV-2-induced autophagy in HUVECs.DENV-2 infection of HUVECs caused increased cellular expressions of LC3 and P62 proteins,which were significantly lowered by treatment with LY294002(a PI3K inhibitor).Conclusion Baicalin inhibits DENV-2 replication in HUVECs and suppresses DENV-2-induced cell autophagy by inhibiting the PI3K/AKT signaling pathway.

Objective To investigate whether activation of mitochondrial acetal dehydrogenase 2(ALDH2)alleviates hypoxic pulmonary hypertension by regulating the SIRT1/PGC-1α signaling pathway.Methods Thirty 8-week-old C57 BL/6 mice were randomized into control,hypoxia,and hypoxia+Alda-1(an ALDH2 activator)group(n=10),and the mice in the latter two groups,along with 10 ALDH2 knockout(ALDH2-/-)mice,were exposed to hypoxia(10%O2,90%N2)with or without daily intraperitoneal injection of Alda-1 for 4 weeks.The changes in right ventricular function and pressure(RVSP)of the mice were evaluated by echocardiography and right ventricular catheter test,and pulmonary artery pressure was estimated based on RVSP.Pulmonary vascular remodeling,right ventricular injury,myocardial α-SMA expression,distal pulmonary arteriole muscle normalization,right ventricular cross-sectional area,myocardial cell hypertrophy,and right cardiac hypertrophy index were assessed with HE staining,immunofluorescence staining and WGA staining,and the expressions of ALDH2,SIRT1,PGC-1α,P16INK4A and P21CIP1 were detected.In pulmonary artery smooth muscle cells with hypoxic exposure,the effect of Alda-1 and EX527 on cell senescence and protein expressions was evaluated using β-galactose staining and Western blotting.Results The wild-type mice with hypoxic exposure showed significantly increased RVSP,right ventricular free wall thickness and myocardial expressions of P16INK4A and P21CIP1,which were effectively lowered by treatment with Alda-1 but further increased in ALDH2-/-mice.In cultured pulmonary artery smooth muscle cells,hypoxic exposure significantly increased senescent cell percentage and cellular expressions of P16INK4A and P21CIP1,which were all lowered by treatment with Alda-1,but its effect was obviously attenuated by EX527 treatment.Conclusion ALDH2 alleviates hypoxia-induced senescence of pulmonary artery smooth muscle cells by upregulating the SIRT1/PGC-1α signaling pathway to alleviate pulmonary hypertension in mice.

Background:Previous studies have proved that neutrophil gelatinase-associated lipocalin(NGAL)plays an important role in the progression of Crohn's disease(CD),and may serve as a potential biomarker for disease activity prediction,severity assessment,treatment response evaluation and prognosis monitoring.However,the diagnostic value of NGAL in perianal fistulizing Crohn's disease(pfCD)is still unclear.Aims:To investigate the serum level of NGAL and its diagnostic value in patients with active pfCD.Methods:A total of 66 patients diagnosed as pfCD from July 2021 to June 2023 at Longhua Hospital,Shanghai University of Traditional Chinese Medicine were enrolled,including 36 active pfCD patients and 30 inactive pfCD patients.The disease activity and perianal fistula activity were assessed by Crohn's disease activity index(CDAI)and perianal disease activity index(PDAI),respectively.Serum NGAL,fecal calprotectin(FC),C-reactive protein(CRP),erythrocyte sedimentation rate(ESR),as well as CDAI score and PDAI score were compared between the active and inactive pfCD patients,and the correlations of NGAL with the other parameters in active pfCD patients were analyzed.ROC curve was drawn to evaluate the values of serum NGAL,FC,CRP and ESR for diagnosis of active pfCD.Results:The serum NGAL,FC,CRP,ESR,CDAI score and PDAI score in active pfCD patients were significantly higher than those in inactive pfCD patients(all P＜0.001).NGAL was positively correlated with FC(r=0.64,P＜0.001),CRP(r=0.55,P＜0.001),ESR(r=0.53,P＜0.001),CDAI score(r=0.59,P＜0.001)and PDAI score(r=0.54,P＜0.001)in active pfCD patients.The optimal cut-off values of NGAL,FC,CRP and ESR were 220.5 μg/L,146.0 μg/g,7.9 mg/L and 23.5 mm/h,respectively,for the diagnosis of active pfCD,and the area under the curve were 0.922(95％CI:0.850-0.995),0.888(95％CI:0.806-0.970),0.853(95％CI:0.763-0.944)and 0.830(95％CI:0.731-0.930),respectively.Conclusions:Serum NGAL level is associated with the disease activity of pfCD,and can be used as a non-invasive biomarker for the clinical diagnosis of active pfCD.

Objective To investigate the effect of type 2 dengue virus(DENV-2)infection on autophagy in human umbilical vein endothelial cells(HUVECs)and the mechanism mediating the inhibitory effect of baicalin against DENV-2 infection.Methods Cultured HUVECs with DENV-2 infection were treated with different concentrations of baicalin,and the changes in autophagy of the cells were detected using transmission electron microscopy.Lyso Tracker Red staining was used to examine pH changes in the lysosomes of the cells,and the expressions of ATG5,beclin-1,LC3,P62,STX17,SNAP29,VAMP8,and PI3K/AKT signaling pathway-related proteins were detected by Western blotting.DENV-2 replication in the cells were evaluated using RT-qPCR.The differentially expressed proteins in DENV-2-infected HUVECs were identified by proteomics screening.Results Treatment with baicalin did not significantly affect the viability of cultured HUVECs.Proteomic studies suggested that the PI3K-AKT pathway played an important role in mediating cell injury induced by DENV-2 infection.The results of RT-qPCR demonstrated that baicalin dose-dependently inhibited DENV-2 replication in HUVECs and produced the strongest inhibitory effect at the concentration of 50 μg/mL.Transmission electron microscopy,Lyso Tracker Red staining,RT-qPCR,and Western blotting all showed significant inhibitory effect of baicalin on DENV-2-induced autophagy in HUVECs.DENV-2 infection of HUVECs caused increased cellular expressions of LC3 and P62 proteins,which were significantly lowered by treatment with LY294002(a PI3K inhibitor).Conclusion Baicalin inhibits DENV-2 replication in HUVECs and suppresses DENV-2-induced cell autophagy by inhibiting the PI3K/AKT signaling pathway.

Objective To investigate whether activation of mitochondrial acetal dehydrogenase 2(ALDH2)alleviates hypoxic pulmonary hypertension by regulating the SIRT1/PGC-1α signaling pathway.Methods Thirty 8-week-old C57 BL/6 mice were randomized into control,hypoxia,and hypoxia+Alda-1(an ALDH2 activator)group(n=10),and the mice in the latter two groups,along with 10 ALDH2 knockout(ALDH2-/-)mice,were exposed to hypoxia(10%O2,90%N2)with or without daily intraperitoneal injection of Alda-1 for 4 weeks.The changes in right ventricular function and pressure(RVSP)of the mice were evaluated by echocardiography and right ventricular catheter test,and pulmonary artery pressure was estimated based on RVSP.Pulmonary vascular remodeling,right ventricular injury,myocardial α-SMA expression,distal pulmonary arteriole muscle normalization,right ventricular cross-sectional area,myocardial cell hypertrophy,and right cardiac hypertrophy index were assessed with HE staining,immunofluorescence staining and WGA staining,and the expressions of ALDH2,SIRT1,PGC-1α,P16INK4A and P21CIP1 were detected.In pulmonary artery smooth muscle cells with hypoxic exposure,the effect of Alda-1 and EX527 on cell senescence and protein expressions was evaluated using β-galactose staining and Western blotting.Results The wild-type mice with hypoxic exposure showed significantly increased RVSP,right ventricular free wall thickness and myocardial expressions of P16INK4A and P21CIP1,which were effectively lowered by treatment with Alda-1 but further increased in ALDH2-/-mice.In cultured pulmonary artery smooth muscle cells,hypoxic exposure significantly increased senescent cell percentage and cellular expressions of P16INK4A and P21CIP1,which were all lowered by treatment with Alda-1,but its effect was obviously attenuated by EX527 treatment.Conclusion ALDH2 alleviates hypoxia-induced senescence of pulmonary artery smooth muscle cells by upregulating the SIRT1/PGC-1α signaling pathway to alleviate pulmonary hypertension in mice.

Natural killer (NK) cells are an important part of the body's innate immune system. As the first line of defense against pathogens, they need to be transformed into a mature state under the control of various cell signaling molecules and transcription factors to play cytotoxic and immune regulatory roles. Under the interaction of activated receptors and inhibitory receptors, NK cells are activated to perform a direct cell killing effect by secreting perforin and granzyme, or indirectly eliminate pathogenic microorganisms in the body by secreting various cytokines, such as type I and type II interferons. These functions of NK cells play a very important role in antiviral and anti-autoimmune diseases, especially in anti-tumor.
Humans ; Killer Cells, Natural ; Interferon-gamma ; Apoptosis ; Autoimmune Diseases ; Cytokines

The α-amino acid ester acyltransferase (SAET) from Sphingobacterium siyangensis is one of the enzymes with the highest catalytic ability for the biosynthesis of l-alanyl-l-glutamine (Ala-Gln) with unprotected l-alanine methylester and l-glutamine. To improve the catalytic performance of SAET, a one-step method was used to rapidly prepare the immobilized cells (SAET@ZIF-8) in the aqueous system. The engineered Escherichia coli (E. coli) expressing SAET was encapsulated into the imidazole framework structure of metal organic zeolite (ZIF-8). Subsequently, the obtained SAET@ZIF-8 was characterized, and the catalytic activity, reusability and storage stability were also investigated. Results showed that the morphology of the prepared SAET@ZIF-8 nanoparticles was basically the same as that of the standard ZIF-8 materials reported in literature, and the introduction of cells did not significantly change the morphology of ZIF-8. After repeated use for 7 times, SAET@ZIF-8 could still retain 67% of the initial catalytic activity. Maintained at room temperature for 4 days, 50% of the original catalytic activity of SAET@ZIF-8 could be retained, indicating that SAET@ZIF-8 has good stability for reuse and storage. When used in the biosynthesis of Ala-Gln, the final concentration of Ala-Gln reached 62.83 mmol/L (13.65 g/L) after 30 min, the yield reached 0.455 g/(L·min), and the conversion rate relative to glutamine was 62.83%. All these results suggested that the preparation of SAET@ZIF-8 is an efficient strategy for the biosynthesis of Ala-Gln.
Escherichia coli/genetics* ; Glutamine ; Zeolites/chemistry* ; Amino Acids

OBJECTIVE: To prepare customized porous silicone orbital implants using embedded 3D printing and assess the effect of surface modification on the properties of the implants. METHODS: The transparency, fluidity and rheological properties of the supporting media were tested to determine the optimal printing parameters of silicone. The morphological changes of silicone after modification were analyzed by scanning electron microscopy, and the hydrophilicity and hydrophobicity of silicone surface were evaluated by measuring the water contact angle. The compression modulus of porous silicone was measured using compression test. Porcine aortic endothelial cells (PAOECs) were co-cultured with porous silicone scaffolds for 1, 3 and 5 days to test the biocompatibility of silicone. The local inflammatory response to subcutaneous porous silicone implants was evaluated in rats. RESULTS: The optimal printing parameters of silicone orbital implants were determined as the following: supporting medium 4% (mass ratio), printing pressure 1.0 bar and printing speed 6 mm/s. Scanning electron microscopy showed that the silicone surface was successfully modified with polydopamine and collagen, which significantly improved hydrophilicity of the silicone surface (P < 0.05) without causing significant changes in the compression modulus (P > 0.05). The modified porous silicone scaffold had no obvious cytotoxicity and obviously promoted adhesion and proliferation of PAOECs (P < 0.05). In rats bearing the subcutaneous implants, no obvious inflammation was observed in the local tissue. CONCLUSION Poprous silicone orbital implants with uniform pores can be prepared using embedded 3D printing technology, and surface modification obviously improves hydrophilicity and biocompatibility of the silicone implants for potential clinical application.
Animals ; Rats ; Swine ; Silicon ; Orbital Implants ; Endothelial Cells ; Porosity ; Silicones ; Printing, Three-Dimensional