BACKGROUND:Traditional methods of urinary tract reconstruction are limited by donor scarcity,high complication rates,and suboptimal functional recovery.Tissue engineering strategies offer new directions in this field.Since the urinary tract is mainly composed of muscle tissue,the key is to find suitable seed cells and efficiently induce them to differentiate into smooth muscle cells.Comparative studies on the efficacy of different smooth muscle cell induction regimens are still lacking. OBJECTIVE:To isolate,culture,and identify human urine-derived stem cells,and to compare the effects of two different induction protocols. METHODS:Human urine-derived stem cells were isolated from urine samples of 11 healthy adult volunteers by multiple centrifugations.Surface markers were identified by flow cytometry.The multi-directional differentiation potential of human urine-derived stem cells was verified through osteogenic and adipogenic differentiation.Differentiation was induced by transforming growth factor-β1 or transforming growth factor-β1 combined with platelet derived growth factor for 14 days.Immunofluorescence staining and western blot assay were employed to compare the expression differences of smooth muscle-specific proteins(α-SMA and SM22). RESULTS AND CONCLUSION:(1)Urine-derived stem cells were successfully isolated from the eight urine samples of healthy people.These cells exhibit a"rice grain"-like morphology and possess a robust proliferative capacity.(2)Urine-derived stem cells exhibited high expression of mesenchymal stem cell surface markers(CD73,CD90,and CD44)and extremely low expression of hematopoietic stem cell surface markers(CD34 and CD45).These cells did not express CD19,CD105,and HLA-DR.(3)After osteogenic and adipogenic differentiation,the formation of calcium nodules and lipid droplets was observed,with positive staining results from Alizarin Red S and Oil Red O staining.(4)After 14 days of smooth muscle induction culture,immunofluorescence staining revealed that the smooth muscle differentiation rate of urine-derived stem cells treated with a combination of transforming growth factor-β1 and platelet derived growth factor was significantly higher compared to those treated with transforming growth factor-β1 alone(P<0.005).(5)After 14 days of smooth muscle induction culture,western blot assay further demonstrated that the expression levels of α-SMA and SM22 in the transforming growth factor-β1/platelet derived growth factor group were significantly elevated compared to those in the transforming growth factor-β1 only group(P<0.005).These findings confirm that urine-derived stem cells can be non-invasively isolated using multiple rounds of centrifugation.Compared with transforming growth factor-β1 alone,the combination of transforming growth factor-β1 and platelet derived growth factor can improve the efficiency of inducing urine-derived stem cells to differentiate into smooth muscle cells.

Objective:To assess the predictive value for the risk of cardiogenic stroke (CS) in patients with paroxysmal atrial fibrillation (PAF) using quantification of left atrial appendage early blood stasis (LAA-BS) signs derived from left atrium-pulmonary vein CT examination.Methods:A retrospective analysis of 187 patients with PAF, who were confirmed to have LAA-BS by left atrium-pulmonary vein CT examinations, was conducted at Second Affiliated Hospital of Nantong University from January 2019 to December 2021. The ratio of LAA-BS CT values to ascending aorta (AA) CT values (HU BS/HU AA) and the ratio of LAA-BS volume to LAA volume (V BS/V LAA) were measured at the peak time of AA enhancement, which were used as characteristic quantitative indicators of LAA-BS. Using the median values of HU BS/HU AA and V BS/V LAA as cut-off points for grouping, the differences between the high-ratio and low-ratio groups were compared in terms of general information, clinical characteristics, and imaging characteristics. All enrolled patients were followed up with the primary outcome event of CS occurrence. The differences in the proportion of CS occurrence between the high-ratio and low-ratio groups were compared. The risk stratification analysis of the occurrence of CS in PAF patients was performed using Kaplan-Meier curves. Additionally, the predictive value of HU BS/HU AA, V BS/V LAA and other imaging indices for the risk of CS occurrence was assessed using Cox proportional risk regression models. Results:The incidence of hypertension and the proportion of patients with atrial fibrillation-stroke risk score (CHA 2DS 2-VASc)≥3 in the high V BS/V LAA group were higher than that in the low V BS/V LAA group, and the difference was statistically significant ( P=0.041, P=0.011). The left atrial volume (LAV) in patients in the low HU BS/HU AA group was greater than in the high HU BS/HU AA group, and the difference was statistically significant ( P=0.040). Kaplan-Meier analysis showed a higher incidence of CS in the low HU BS/HU AA group than in the high HU BS/HU AA group ( P=0.012). Similarly, the high V BS/V LAA group had a higher incidence of CS compared with the low V BS/V LAA group ( P=0.019). Subgroup analysis revealed a significantly higher incidence of CS in the subgroup with low HU BS/HU AA and high V BS/V LAA compared to other subgroups (all P<0.05). The Cox proportional hazards regression model, adjusting for confounding factors, identified low HU BS/HU AA and high V BS/V LAA as independent risk factors for CS occurrence in PAF patients ( P=0.005 and P=0.029). Conclusion:The HU BS/HU AA and V BS/V LAA quantified using left atrium-pulmonary vein CT imaging are predictive factors for CS occurrence in patients with PAF. These ratios synergistically contribute to the risk assessment of CS.

Debates persist regarding the efficacy and safety of azvudine, particularly its real-world outcomes. This study involved patients aged ≥60 years who were admitted to 25 hospitals in mainland China with confirmed SARS-CoV-2 infection between December 1, 2022, and February 28, 2023. Efficacy outcomes were all-cause mortality during hospitalization, the proportion of patients discharged with recovery, time to nucleic acid-negative conversion (T _NANC), time to symptom improvement (T _SI), and time of hospital stay (T _HS). Safety was also assessed. Among the 5884 participants identified, 1999 received azvudine, and 1999 matched controls were included after exclusion and propensity score matching. Azvudine recipients exhibited lower all-cause mortality compared with controls in the overall population (13.3% vs. 17.1%, RR, 0.78; 95% CI, 0.67-0.90; P = 0.001) and in the severe subgroup (25.7% vs. 33.7%; RR, 0.76; 95% CI, 0.66-0.88; P < 0.001). A higher proportion of patients discharged with recovery, and a shorter T _NANC were associated with azvudine recipients, especially in the severe subgroup. The incidence of adverse events in azvudine recipients was comparable to that in the control group (2.3% vs. 1.7%, P = 0.170). In conclusion, azvudine showed efficacy and safety in older patients hospitalized with COVID-19 during the SARS-CoV-2 omicron wave in China.

ObjectiveTo investigate the effects and potential mechanisms of morning and evening fasting on postprandial lipid responses, a post hoc analysis based on a crossover randomized controlled trial was conducted to assess the effects of different fasting strategies on postprandial lipid metabolism in community residents in Shanghai. MethodsA total of 23 participants took part in a randomized crossover trial involving two intervention days: morning fasting and evening fasting, with a washout period of 6 days between intervention days. Two-way analysis of variance was used to test the differences in total cholesterol (TC), triglycerides (TG), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), and the relative expression of circadian clock genes before and after the next meal under fasting. Wilcoxon rank sum tests were used to analyze the different metabolites between the two groups. Principal component analysis and Orthogonal partial least squares-discriminant analysis were conducted to evaluate the ability of metabolites to differentiate between morning fasting and evening fasting and identify the important differential metabolites. After adjusting for age, sex, and BMI, a partial correlation analysis was performed to identify metabolites associated with plasma lipids. In addition, important metabolites associated with plasma lipids were computed by pathway enrichment analysis. ResultsAfter evening fasting intervention, fasting TG level [(0.37±0.29) vs (0.27±0.18)] mmol·L^-1, fasting and postprandial change values in TC [(2.74±0.47) vs (2.51±0.27)] mmol·L^-1 and LDL-C [(1.32±0.38) vs (0.99±0.27)] mmol·L^-1 were significantly lower than those after morning fasting (P<0.05). While, change values of fasting LDL-C [(0.89±0.37) vs (1.14±0.37)] mmol·L^-1 and TG [(1.14±0.19) vs (1.28±0.17)] mmol·L^-1 were significantly higher than those after morning fasting intervention (P<0.05). After fasting intervention, the relative expression of AMPK, CRY1, CLOCK, MTNR1B, AANAT, and ASMT was correlated with the amount of plasma lipid changes (P<0.05). Specifically, CLOCK and AANAT were upregulated following evening fasting and downregulated after morning fasting. Among the 217 important differential metabolites, 111 were correlated with plasma lipids, and which were primarily enriched in the cysteine and methionine metabolism pathways (P<0.05). ConclusionCompared to morning fasting, evening fasting was more effective in improving postprandial lipid responses, indicating that an evening fasting window during intermittent fasting could be conducive to cardiovascular disease prevention in adults. Meanwhile, it is suggested that morning and evening fasting may affect lipid responses through circadian rhythm oscillations and the cysteine and methionine metabolism pathways.

Knee osteoarthritis(KOA)and osteoporosis(OP)"co-morbidity"is a chronic progressive disease,the incidence of which is higher in recent years.Exosomes have intercellular messenger functions,which play an important role in the development of KOA and OP"co-morbidity".This article recognized the"co-morbidity"between KOA and OP from the perspective of modern medicine and the"tendon","bone","liver"and"kidney"in TCM theories,systematically reviewed the indirect regulation of KOA and OP"co-morbidity"through exosomes,and explored the possible mechanisms of exosome-based treatment of KOA and OP"co-morbidity"through Chinese materia medica in terms of inflammation,MAPK pathway,bone remodeling and cartilage metabolism,and mitochondrial autophagy,with the aim to investigate the possible mechanisms of KOA and OP"co-morbidity",and provide new ideas for the treatment of KOA and OP"co-morbidity".

Knee osteoarthritis(KOA)and osteoporosis(OP)"co-morbidity"is a chronic progressive disease,the incidence of which is higher in recent years.Exosomes have intercellular messenger functions,which play an important role in the development of KOA and OP"co-morbidity".This article recognized the"co-morbidity"between KOA and OP from the perspective of modern medicine and the"tendon","bone","liver"and"kidney"in TCM theories,systematically reviewed the indirect regulation of KOA and OP"co-morbidity"through exosomes,and explored the possible mechanisms of exosome-based treatment of KOA and OP"co-morbidity"through Chinese materia medica in terms of inflammation,MAPK pathway,bone remodeling and cartilage metabolism,and mitochondrial autophagy,with the aim to investigate the possible mechanisms of KOA and OP"co-morbidity",and provide new ideas for the treatment of KOA and OP"co-morbidity".

Objective:To assess the predictive value for the risk of cardiogenic stroke (CS) in patients with paroxysmal atrial fibrillation (PAF) using quantification of left atrial appendage early blood stasis (LAA-BS) signs derived from left atrium-pulmonary vein CT examination.Methods:A retrospective analysis of 187 patients with PAF, who were confirmed to have LAA-BS by left atrium-pulmonary vein CT examinations, was conducted at Second Affiliated Hospital of Nantong University from January 2019 to December 2021. The ratio of LAA-BS CT values to ascending aorta (AA) CT values (HU BS/HU AA) and the ratio of LAA-BS volume to LAA volume (V BS/V LAA) were measured at the peak time of AA enhancement, which were used as characteristic quantitative indicators of LAA-BS. Using the median values of HU BS/HU AA and V BS/V LAA as cut-off points for grouping, the differences between the high-ratio and low-ratio groups were compared in terms of general information, clinical characteristics, and imaging characteristics. All enrolled patients were followed up with the primary outcome event of CS occurrence. The differences in the proportion of CS occurrence between the high-ratio and low-ratio groups were compared. The risk stratification analysis of the occurrence of CS in PAF patients was performed using Kaplan-Meier curves. Additionally, the predictive value of HU BS/HU AA, V BS/V LAA and other imaging indices for the risk of CS occurrence was assessed using Cox proportional risk regression models. Results:The incidence of hypertension and the proportion of patients with atrial fibrillation-stroke risk score (CHA 2DS 2-VASc)≥3 in the high V BS/V LAA group were higher than that in the low V BS/V LAA group, and the difference was statistically significant ( P=0.041, P=0.011). The left atrial volume (LAV) in patients in the low HU BS/HU AA group was greater than in the high HU BS/HU AA group, and the difference was statistically significant ( P=0.040). Kaplan-Meier analysis showed a higher incidence of CS in the low HU BS/HU AA group than in the high HU BS/HU AA group ( P=0.012). Similarly, the high V BS/V LAA group had a higher incidence of CS compared with the low V BS/V LAA group ( P=0.019). Subgroup analysis revealed a significantly higher incidence of CS in the subgroup with low HU BS/HU AA and high V BS/V LAA compared to other subgroups (all P<0.05). The Cox proportional hazards regression model, adjusting for confounding factors, identified low HU BS/HU AA and high V BS/V LAA as independent risk factors for CS occurrence in PAF patients ( P=0.005 and P=0.029). Conclusion:The HU BS/HU AA and V BS/V LAA quantified using left atrium-pulmonary vein CT imaging are predictive factors for CS occurrence in patients with PAF. These ratios synergistically contribute to the risk assessment of CS.

ObjectiveTo understand the relative bioavailability （RBA） of cadmium in different aquatic products. Based on the consideration of the gender differences and the relative bioavailability of cadmium in different foods， the physiologically based toxicokinetic （PBTK） model of cadmium was further optimized and verified. The correlation between internal and external exposure in quantitative risk assessment of food safety was optimized， and the provisional tolerable daily intake （PTDI） value of cadmium was derived. MethodsThe relative bioavailability of cadmium in different aquatic products was determined in four-week-old Balb/c female mice. The PBTK model of cadmium metabolism was optimized by ABC-MCMC method using combined internal and external exposure data of cadmium in Shanghai residents， and the PTDI was calculated accordingly. ResultsExcept for scallops and squid， the RBA of aquatic samples was less than 1， indicating that the absorption rate of cadmium in aquatic products was lower than that of cadmium chloride. The higher RBA of squid and scallop may be due to the presence of cadmium in the visceral organs， which is conducive to cadmium absorption and its higher concentration of cadmium. Frying at the temperature less than 160 ℃ reduced cadmium absorption but may increase cadmium absorption at the temperature greater than 160 ℃. The optimized model parameters converged well and the model could reasonably estimate urinary cadmium level according to the external exposure of cadmium. The PTDI value was0.466 4 μg·（kg·day）^-1 according to the optimized single-chamber model. ConclusionThe relative bioavailability of cadmium in different foods varies greatly， except for squid and scallops， RBA is less than 1， and cooking processing will affect the RBA of food. The construction of the PBTK model did not only consider the effects of gender differences on cadmium metabolism， but also included the relative bioavailability data to optimize and adjust the correlation coefficient of absorption rate. Compared with the model without RBA adjustment， the adjusted model has enhanced the ability to predict urinary cadmium level， which provides a new， more accurate method for the risk assessment of food safety.

Objective:To investigate the effect of low-dose ketamine on neuroinflammation and microcirculation in mice with traumatic brain injury (TBI).Methods:Sixty adult male C57BL/6 mice, weighing 22-28 g, were randomly divided into sham-operated group, TBI group, Sham+ketamine group, and TBI+ketamine group ( n=15). A controlled cortical impingement (CCI) method was used to establish TBI models in the later 2 groups. Sham+ketamine group and TBI+ketamine group were intraperitoneally injected with 30 mg/kg ketamine once daily for 3 d at 30 min after TBI; sham-operated group and TBI group were intraperitoneally injected same amount of saline at the same time points. Cerebral cortical blood flow in 6 mice from each group was measured by laser speckle contrast imaging (LSCI) before, immediately after, 30 min after, 1 d after and 3 d after modeling, respectively. Three d after modeling, immunohistochemical staining and immunofluorescent double label staining were used to detect the nuclear translocation of microglia markers, ionized calcin-antibody-1 (Iba-1) and nuclear factor (NF)-κB p65 in damaged cortical brain tissues in 6 mice from each group. The remaining 3 mice in each group were sacrificed and tissue plasma was extracted 3 d after modeling; levels of NF-κB p65, phosphorylated (p)-NF-κB p65, p-IκB and inducible nitric oxide synthase (iNOS) in cortical brain tissues were detected by Western blotting. Expressions of tumor necrosis factor-α (TNF-α), interleukin-1-β (IL-1β) and interleukin-6 (IL-6), iNOS, reactive oxygen species (ROS) and reactive nitrogen species (RNS) in cortical brain tissues were detected by ELISA. Results:LSCI indicated that, 3 d after modeling, relative blood flow in local cerebral microcirculation of TBI+ketamine group was significantly increased compared with that of TBI group ( P<0.05). Immunohistochemical staining indicated that compared with the sham-operated group and Sham+ketamine group, the TBI group and TBI+ketamine group had significantly increased number of Iba-1 positive cells in the cerebral cortex ( P<0.05); compared with the TBI group, the TBI+ketamine group had significantly decreased number of Iba-1 positive cells ( P<0.05). ELISA indicated that compared with the sham-operated group and Sham+ketamine group, the TBI group and TBI+ketamine group had significantly increased expressions of TNF-α, IL-1β, IL-6, iNOS, ROS and RNS in damaged cortical brain tissues ( P<0.05); compared with the TBI group, the TBI+ ketamine group had significantly decreased expressions of TNF-α, IL-1β, IL-6, iNOS, ROS and RNS in damaged cortical brain tissues ( P<0.05). Immunofluorescent double label staining indicated obviously inhibited NF-κB p65 nuclear translocation in TBI+ketamine group when it was compared with TBI group. Western blotting indicated that compared with the sham-operated group and Sham+ketamine group, the TBI+ketamine group had significantly increased iNOS, NF-κB p65, p-NF-κB p65 and P-IκB protein expressions in damaged cortical brain tissues ( P<0.05); compared with the TBI group, the TBI+ketamine group had significantly decreased protein expressions of iNOS, NF-κB p65, p-NF-κB p65 and p-IκB in damaged cortical brain tissues ( P<0.05). Conclusion:Low-dose ketamine reduces neuroinflammation and improves cerebral microcirculatory blood flow after open TBI, whose mechanism may be related to inhibition of microglia NF-κB/iNOS pathway.

To investigate the effect of mitochondrial division inhibitor 1(Mdivi-1)on imiquimod(IMQ)-induced psoriasis-like skin inflammation in mice and its mechanism,female 8-week-old C57BU6 mice were recruited and randomly divided into control group,IMQ model group,IMQ+Mdivi-1 experiment group.IMQ was used to induce the psoriasis-like skin inflammation model in mice.The mice in the experiment group were injected intraperitoneally(i.p.)with Mdivi-1,and the mice in the control group and model group were injected with the same volume of solvent.The mice were sacrificed on the 7th day for sampling.Psoriasis area and severity index(PASI)score was used to evaluate the severity of skin lesions in each group;the reactive oxygen species(R0S)content in skin tissue was detected by fluorescence staining of frozen section;HE staining was used to observe the histomorphologic change of skin lesions;immunohistochemical staining was used to detect the expression of dynamin-related protein 1(Drp1)in the skin of mice;Western blot was used to detect the protein levels of Drp1,NLRP3 and IL-1β in the skin tissues of mice in each group;and the expressions of IL-17A and IL-18 in mouse serum were detected by ELISA.Data showed that the model group had typical psoriatic lesions such as erythema,scale and thickening,and the Mdivi-1 group demonstrated obvious reduction of the lesions.The PASI score of the experiment group was significantly lower than that of the model group.HE staining indicated that the epidermal thickness of the back skin in the treatment group was significantly lower than that in the model group,and Munro microabscess was significantly reduced.R0S fluorescence staining indicated that ROS content in the experiment group was significantly lower than that in the model group;immunohistochemical results showed that the expression of Drp1 protein in the experiment group was significantly lower than that in the model group;Western blot results showed that the expression levels of Drp1,NLRP3 and IL-1 β in the experiment group were significantly lower than those in the model group;ELISA results indicated that the expressions of IL-17A and IL-18 in serum of mice in the experiment group were lower than those in the model group.Taken together,Mdivi-1 can reduce mitochondrial damage and ROS production by inhibiting the expression of Drp1,thereby reducing the production of NLRP3 inflammasome,down-regulating IL-1 β,IL-18 and IL-17A,and alleviating the IMQ-induced psoriasis-like skin inflammation in mice.