BACKGROUND:The mesenchymal stem cell secretome contains bioactive substances,cytokines,and growth factors.Three-dimensional cell culture can regulate the secretion of these components,potentially enhancing the ability to promote injury repair.OBJECTIVE:To investigate the repair effect of three-dimensional cultured human umbilical cord mesenchymal stem cell secretome on skin injuries in mice.METHODS:Human umbilical cord mesenchymal stem cells were cultured in conventional two-dimensional culture dishes and 96-well U-bottom cell culture plates,from which their secretory components were subsequently collected.The expression of skin damage repair related secretory factors in umbilical cord mesenchymal stem cells was analyzed using RT-qPCR.The protein expression level of skin damage repair related factors in umbilical cord mesenchymal stem cell secretome was detected using enzyme-linked immunosorbent assay.The potential of human umbilical cord mesenchymal stem cell secretome to repair vascular injuries was evaluated using an immortalized human umbilical vein endothelial cell migration model.A mouse skin injury model was established,and the human umbilical cord mesenchymal stem cell secretome was injected subcutaneously.Repair effects on skin injury were assessed through wound healing rates and histopathological analysis.RESULTS AND CONCLUSION:(1)After three days of cultivation,human umbilical cord mesenchymal stem cells cultured in two dimensions exhibited a fibroblast-like,swirling growth pattern,whereas three-dimensional culture led to the formation of uniform microspheres.(2)Compared with two-dimensional culture,three-dimensional culture significantly increased the mRNA expression of transforming growth factor β and basic fibroblast growth factor in human umbilical cord mesenchymal stem cells.(3)Compared with two-dimensional culture,three-dimensional cultured human umbilical cord mesenchymal stem cell secretome significantly enhanced the protein expression of vascular endothelial growth factor,interleukin-10,and granulocyte-macrophage colony-stimulating factor in the human umbilical cord mesenchymal stem cell secretome.(4)Compared with two-dimensional culture,three-dimensional cultured human umbilical cord mesenchymal stem cell secretome significantly promoted the migration of immortalized human umbilical cord mesenchymal stem cells.(5)Compared with the untreated control group and the two-dimensional cultured human umbilical cord mesenchymal stem cell secretome,the three-dimensional cultured human umbilical cord mesenchymal stem cell secretome can significantly accelerate the skin wound healing rate and wound skin structure remodeling in mice.These results indicate that three-dimensional culture can enhance the expression of paracrine factors of human umbilical cord mesenchymal stem cells,and their secretome can significantly promote the repair of mouse skin damage.

ObjectiveTo investigate the molecular mechanism by which Yifei Xuanfei Jiangzhuo prescription regulates the nuclear factor-κB （NF-κB）/NOD-like receptor protein 3 （NLRP3） signaling pathway to improve neuronal function in vascular dementia （VaD） rats. MethodsA VaD model was established by intermittently clamping the bilateral common carotid arteries （CCA） combined with bilateral vascular occlusion （2-VO）. Eighty-four SD rats were randomly divided into a blank group， sham group， model group， piracetam group （0.2 g·kg^-1）， and low-， medium-， and high-dose Yifei Xuanfei Jiangzhuo prescription groups （6.09， 12.18， and 24.36 g·kg^-1）. Drug administration began on day 7 after surgery， once daily for 28 consecutive days. Behavioral experiments were used to evaluate learning and spatial memory. Hematoxylin-eosin （HE） staining was applied to observe pathological morphological changes in the CA1 region of the hippocampus. Transmission electron microscopy was used to examine the ultrastructure of hippocampal neurons. Terminal deoxynucleotidyl transferase dUTP nick end labeling （TUNEL） was used to detect neuronal apoptosis in the CA1 region. Immunohistochemistry was performed to determine the positive expression rate of neuronal nuclear antigen （NeuN）. Immunofluorescence single staining was used to assess nuclear expression of NF-κB p65 in brain tissue. Western blot was used to detect the protein expression levels of inhibitor of κB kinase （IKK）， NF-κB p65， NLRP3， Caspase-1， apoptosis-associated speck-like protein （ASC）， and interleukin-1β （IL-1β）. ResultsCompared with the blank group， the model group showed a significant reduction in platform-crossing frequency （P0.01）， aggravated hippocampal injury， a significant increase in neuronal apoptosis （P0.05）， decreased NeuN positivity in the CA1 region （P0.05）， increased nuclear expression of NF-κB p65 （P0.05）， and significantly elevated expression of p-IKK， p-NF-κB p65， NLRP3， cleaved Caspase-1， ASC， and cleaved IL-1β （P0.05）. Compared with the model group， all drug-treated groups improved learning and spatial memory in VaD rats， alleviated hippocampal pathological injury and neuronal apoptosis， and protected neuronal ultrastructure. Yifei Xuanfei Jiangzhuo prescription at doses of 12.18 and 24.36 g·kg^-1 reduced hippocampal expression levels of p-IKK， p-NF-κB p65， NLRP3， Caspase-1， ASC， and cleaved IL-1β in VaD rats （P0.05）， showing dose-dependent inhibition of the NF-κB/NLRP3 signaling pathway. ConclusionYifei Xuanfei Jiangzhuo prescription may exert neuroprotective effects by regulating the NF-κB/NLRP3 signaling pathway， thereby reducing neuroinflammation and inhibiting hippocampal neuronal apoptosis.

ObjectiveTo investigate the molecular mechanism by which Yifei Xuanfei Jiangzhuo prescription regulates the nuclear factor-κB （NF-κB）/NOD-like receptor protein 3 （NLRP3） signaling pathway to improve neuronal function in vascular dementia （VaD） rats. MethodsA VaD model was established by intermittently clamping the bilateral common carotid arteries （CCA） combined with bilateral vascular occlusion （2-VO）. Eighty-four SD rats were randomly divided into a blank group， sham group， model group， piracetam group （0.2 g·kg^-1）， and low-， medium-， and high-dose Yifei Xuanfei Jiangzhuo prescription groups （6.09， 12.18， and 24.36 g·kg^-1）. Drug administration began on day 7 after surgery， once daily for 28 consecutive days. Behavioral experiments were used to evaluate learning and spatial memory. Hematoxylin-eosin （HE） staining was applied to observe pathological morphological changes in the CA1 region of the hippocampus. Transmission electron microscopy was used to examine the ultrastructure of hippocampal neurons. Terminal deoxynucleotidyl transferase dUTP nick end labeling （TUNEL） was used to detect neuronal apoptosis in the CA1 region. Immunohistochemistry was performed to determine the positive expression rate of neuronal nuclear antigen （NeuN）. Immunofluorescence single staining was used to assess nuclear expression of NF-κB p65 in brain tissue. Western blot was used to detect the protein expression levels of inhibitor of κB kinase （IKK）， NF-κB p65， NLRP3， Caspase-1， apoptosis-associated speck-like protein （ASC）， and interleukin-1β （IL-1β）. ResultsCompared with the blank group， the model group showed a significant reduction in platform-crossing frequency （P0.01）， aggravated hippocampal injury， a significant increase in neuronal apoptosis （P0.05）， decreased NeuN positivity in the CA1 region （P0.05）， increased nuclear expression of NF-κB p65 （P0.05）， and significantly elevated expression of p-IKK， p-NF-κB p65， NLRP3， cleaved Caspase-1， ASC， and cleaved IL-1β （P0.05）. Compared with the model group， all drug-treated groups improved learning and spatial memory in VaD rats， alleviated hippocampal pathological injury and neuronal apoptosis， and protected neuronal ultrastructure. Yifei Xuanfei Jiangzhuo prescription at doses of 12.18 and 24.36 g·kg^-1 reduced hippocampal expression levels of p-IKK， p-NF-κB p65， NLRP3， Caspase-1， ASC， and cleaved IL-1β in VaD rats （P0.05）， showing dose-dependent inhibition of the NF-κB/NLRP3 signaling pathway. ConclusionYifei Xuanfei Jiangzhuo prescription may exert neuroprotective effects by regulating the NF-κB/NLRP3 signaling pathway， thereby reducing neuroinflammation and inhibiting hippocampal neuronal apoptosis.

BACKGROUND:The mesenchymal stem cell secretome contains bioactive substances,cytokines,and growth factors.Three-dimensional cell culture can regulate the secretion of these components,potentially enhancing the ability to promote injury repair.OBJECTIVE:To investigate the repair effect of three-dimensional cultured human umbilical cord mesenchymal stem cell secretome on skin injuries in mice.METHODS:Human umbilical cord mesenchymal stem cells were cultured in conventional two-dimensional culture dishes and 96-well U-bottom cell culture plates,from which their secretory components were subsequently collected.The expression of skin damage repair related secretory factors in umbilical cord mesenchymal stem cells was analyzed using RT-qPCR.The protein expression level of skin damage repair related factors in umbilical cord mesenchymal stem cell secretome was detected using enzyme-linked immunosorbent assay.The potential of human umbilical cord mesenchymal stem cell secretome to repair vascular injuries was evaluated using an immortalized human umbilical vein endothelial cell migration model.A mouse skin injury model was established,and the human umbilical cord mesenchymal stem cell secretome was injected subcutaneously.Repair effects on skin injury were assessed through wound healing rates and histopathological analysis.RESULTS AND CONCLUSION:(1)After three days of cultivation,human umbilical cord mesenchymal stem cells cultured in two dimensions exhibited a fibroblast-like,swirling growth pattern,whereas three-dimensional culture led to the formation of uniform microspheres.(2)Compared with two-dimensional culture,three-dimensional culture significantly increased the mRNA expression of transforming growth factor β and basic fibroblast growth factor in human umbilical cord mesenchymal stem cells.(3)Compared with two-dimensional culture,three-dimensional cultured human umbilical cord mesenchymal stem cell secretome significantly enhanced the protein expression of vascular endothelial growth factor,interleukin-10,and granulocyte-macrophage colony-stimulating factor in the human umbilical cord mesenchymal stem cell secretome.(4)Compared with two-dimensional culture,three-dimensional cultured human umbilical cord mesenchymal stem cell secretome significantly promoted the migration of immortalized human umbilical cord mesenchymal stem cells.(5)Compared with the untreated control group and the two-dimensional cultured human umbilical cord mesenchymal stem cell secretome,the three-dimensional cultured human umbilical cord mesenchymal stem cell secretome can significantly accelerate the skin wound healing rate and wound skin structure remodeling in mice.These results indicate that three-dimensional culture can enhance the expression of paracrine factors of human umbilical cord mesenchymal stem cells,and their secretome can significantly promote the repair of mouse skin damage.

Transcription factor PU.1, as a core member of the ETS family, plays a pivotal role in the multi-lineage differentiation of hematopoietic stem cells, particularly in the regulation of erythroid differentiation. PU.1 orchestrates the process of hematopoietic stem cell differentiation towards erythroid cells by modulating the transcription of lineage-determining factors and interacting with other key transcription factors in a fine-tuned manner. PU.1 plays an irreplaceable role in the development and function of red blood cells, with its abnormal expression closely related to the occurrence and progression of various blood diseases, including leukemia, myelodysplastic syndromes, and various types of anemia. This article comprehensively analyzes the functional roles and molecular mechanisms of PU.1 in various stages of erythroid differentiation, as well as its potential roles in related blood diseases. This review not only deepens our understanding of the mechanism by which PU.1 regulates erythroid differentiation, but also provides theoretical grounds for blood disease therapies based on PU.1.
Humans ; Proto-Oncogene Proteins/genetics* ; Trans-Activators/genetics* ; Cell Differentiation/physiology* ; Hematologic Diseases/physiopathology* ; Erythroid Cells/cytology* ; Animals ; Erythropoiesis/physiology*

Objective:To evaluate the outcomes of stroke treatment in neurology and rehabilitation departments using the International Classification of Functioning, Disability and Health′s Rehabilitation Set (ICF-RS).Method:The functional status of stroke survivors before and after treatment in the neurology and rehabilitation departments was evaluated using the ICF-RS (30 items). Each patient′s functioning was graded (normal, mild, moderate or severe dysfunction) by an experienced clinical evaluator and also by the intelligent evaluation model in the ICF-RS app. Medical expenditure data during hospitalization were extracted from the hospital′s case records. Rank sum tests compared the functional changes in a patient before and after their rehabilitation. Kappa coefficients were computed to evaluate the consistency of the functional grades assigned by the evaluators and the app.Results:Before the rehabilitation treatment, all 30 items of the ICF-RS were abnormal for all of the patients from the neurology and rehabilitation departments. After their rehabilitation treatment, 25 items had improved significantly for the neurology patients and 8 had improved significantly for those from the rehabilitation department. After their rehabilitation treatment, the average functional improvement among the neurology patients was 25%. For the rehabilitation patients it was 13%. The total expenditure for every 1% improvement in function was Y977 for the neurology patients (including Y143 for rehabilitation) and Y1, 481 for the rehabilitation patients (including Y862 of actual rehabilitation). The proportions of rehabilitation expenditure were thus 14% and 58% respectively. The kappa coefficients quantifying overall consistency were both greater than 0.8.Conclusion:The national standard ICF-RS can be used to evaluate functional changes, rehabilitation efficacy and the composition of stroke patients′ medical expenditures in the early stage of neurology and the recovery period in the rehabilitation department. The consistency of the functional level evaluations between the app and human evaluators is good.

Objective:To investigate the changes in accommodative facility after implantation of V4c implantable collamer lens (ICL) using the intelligent flipper (iFLIP).Methods:A serial case-control study was conducted.Forty patients (80 eyes) who underwent ICL implantation of V4c for myopia correction and completed the follow-up were enrolled at the Affiliated Hospital of Guizhou Medical University from April 2022 to April 2023.Monocular and binocular accommodative facility, adjustment time, and relaxation time were measured with iFLIP before operation and at 1 week, 1 month, 3 months, and 6 months after operation.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of the Affiliated Hospital of Guizhou Medical University (No.2023-983).Written informed consent was obtained from each subject.Results:Monocular and binocular uncorrected visual acuity at 1 week, 1 month, 3 months and 6 months after surgery were significantly improved compared with before surgery, and the differences were statistically significant (all P<0.05).Monocular and binocular postoperative accommodative facility gradually increased over time, and the increase gradually decreased and gradually stabilized.Before operation and 1 week, 1 month, 3 months, and 6 months after operation, the monocular accommodative facility was (7.99±3.42), (10.19±4.25), (12.03±4.24), (13.10±4.66) and (13.64±4.40)cpm, and the binocular accommodative facility was (9.01±3.63), (9.56±3.38), (11.58±4.00), (13.31±3.64), and (14.03±3.72)cpm, respectively, with statistically significant overall differences ( F=24.02, 14.46; both P<0.001).Monocular accommodative facility was higher than before surgery at each time point after surgery, and was higher than 1 week after surgery at 1, 3, and 6 months after surgery, with statistically significant differences (all P<0.05).Binocular accommodative facility was higher at 1, 3, and 6 months after surgery than that before surgery and 1 week after surgery, and higher at 3 and 6 months after surgery than that at 1 month after surgery, with statistically significant differences (all P<0.05).With the extension of postoperative time, the monocular and binocular adjustment time and relaxation time gradually shortened, and the shortening gradually decreased and stabilized.Monocular adjustment time and relaxation time were shorter than before surgery at all time points after surgery.The monocular adjustment time was shorter at 1, 3 and 6 months after surgery than that at 1 week after surgery and shorter at 3 and 6 months after surgery than that at 1 month after surgery, and the monocular relaxation time was shorter at 3 and 6 months after surgery than that at 1 week after surgery, and the differences were statistically significant (all P<0.05).The binocular adjustment time was shorter at 1, 3 and 6 months after surgery than before surgery and at 1 week after surgery, shorter at 3 and 6 months after surgery than at 1 month after surgery and the binocular relaxation time was shorter at all time points after surgery than that before surgery, showing statistically significant differences (all P<0.05). Conclusions:After ICL implantation, uncorrected visual acuity, accommodative facility, adjustment time and relaxation time all improve throughout the preoperative period and eventually stabilize.

Objective:To study genotype analysis and influence factors analysis of hepatitis B virus(HBV)deoxyribonucleic acid(DNA)positive samples of negative hepatitis B surface antigen(HBsAg)donors in blood donors without compensation in Nan'an area.Methods:A total of 62,000 HBsAg fast-screened blood donor samples from January 2021 to June 2024 were selected as the study objects.HBV infection,viral load and genotype of HBV DNA positive samples,single factor and influencing factor of HBV DNA positive samples were detected and analyzed.Results:Among the 62000 HBsAg fast-screened blood donor samples,the results were negative by HBsAg screening,moreover,285 cases(0.46％)were HBsAg positive and 61715 cases(99.54％)were negative.NAT positive 180 cases and 118 cases were positive for HBV DNA.The HBV DNA positive rate of HBsAg negative donors was 0.19％(118/62000).HBV DNA of the viral load of positive specimens with＜20 IU/mL was higher than those with 20 to 99 IU/mL and ≥ 100 IU/mL(P＜0.05).HBV DNA of C style of the genotype of positive specimens was higher than B style and unsamples(P＜0.05).Univariate analysis found that HBV DNA positivity was mainly related to the history of intravenous drug use and risky sexual behavior,as well as the history of hepatitis B vaccination and blood transfusion(P＜0.05).According to the Logistic regression analysis,history of intravenous drug use and risky sexual behavior,and blood transfusion were risk factors for positive HBV DNA(OR＞1,P＜0.05),vaccination of hepatitis B vaccine was a protective factor for HBV DNA positivity(OR＜1,P＜0.05).Conclusion:The proportion of HBsAg in Nan'an area is relatively high,but there are still a small number of positive samples,the main genotype was type C,and history of intravenous drug use and risky risk behaviors,as well as history of blood transfusion were all risk factors for positive HBV DNA,vaccination of hepatitis B vaccine is protective factor for HBV DNA positivity.

Objective:To investigate the MRI characteristics of ductal lesions with microcalcifications detected by mammography,and to evaluate the value of MRI in the differential diagnosis of benign and malignant ductal lesions.Methods:The clinical data of 112 patients with pathologically confirmed ductal lesions with microcalcifications detected by mammography in our hospital from January 2022 to June 2024 were retrospectively analyzed.All patients underwent mammography and MRI examinations.MRI morphological features,dynamic enhancement characteristics,diffusion-weighted imaging(DWI)findings,and apparent diffusion coefficient(ADC)values were analyzed and compared with pathological results.The diagnostic performance of various MRI features was analyzed using ROC curves.Results:Among the 112 patients,there were 47 benign lesions and 65 malignant lesions.Malignant lesions were more likely to present with irregular morphology,heterogeneous T2 signal,lobulation,and spiculation(P＜0.05),while benign lesions were more likely to present with round or oval shapes and homogeneous T2 signal(P＜0.05).Malignant lesions predominantly showed fast rise-fast washout(Washout type)enhancement curves(58.5％),while benign lesions predominantly showed persistent rise(Progressive type)enhancement curves(55.3％).The ADC values of malignant lesions(0.92±0.21× 10-3 mm2/s)were significantly lower than those of benign lesions(1.45±0.28× 10-3 mm2/s)(P＜0.001).The combined application of morphological features,dynamic enhancement curves,and ADC values had the highest diagnostic performance,with a sensitivity of 92.3％,specificity of 89.4％,and accuracy of 91.1％.Conclusion:Multi-parameter MRI can comprehensively evaluate ductal lesions with microcalcifications detected by mammography.The combined application of morphological features,dynamic enhancement curve types,and ADC values can effectively improve the accuracy of differential diagnosis between benign and malignant lesions,providing important basis for clinical treatment decisions.