BACKGROUND:Treadmill training is one of the effective ways to promote the recovery of motor function after spinal cord injury.Treadmill training can promote neurogenesis,but the effect of different intensities of treadmill training on the activation of endogenous stem cells is still unclear. OBJECTIVE:To analyze the activation effect of different intensities of treadmill training on endogenous neural stem cells in the spinal cord of mice after spinal cord injury. METHODS:Fifty female C57BL/6J mice were divided into control group,spinal cord injury group,low-,moderate-,and high-intensity exercise groups with 10 mice in each group by random number table method.T10 segment spinal cord injury model was constructed by the clamp method in spinal cord injury group,low-,moderate-,and high-intensity exercise groups.On day 7 after spinal cord injury,mice in the low-,moderate-,and high-intensity exercise groups were respectively trained on the treadmill with corresponding intensity,3 times/d,10 min/times,6 times a week for 28 consecutive days.At 3,7,14,21,and 28 days after treadmill training,the hind limb motor function was evaluated by BMS score.At 28 days after treadmill training,the spinal cord tissue of the injured area was obtained,and the expression of epidermal growth factor receptor,glial fibrillary acidic protein,and 5-Ethynyl-2'-deoxyuridine(EdU),a proliferative marker,was detected.Hematoxylin-eosin staining was used to observe the morphology of spinal cord. RESULTS AND CONCLUSION:(1)The BMS score of mice in the spinal cord injury group was lower than that in the control group(P<0.05).With the extension of treadmill training time,the BMS scores of mice with spinal cord injury gradually increased,and the BMS scores of mice in moderate-intensity exercise group on days 14 and 21 after treadmill training were higher than those in spinal cord injury group and low-and high-intensity exercise groups(P<0.05).The BMS score of mice in moderate-and high-intensity exercise group was higher than that in spinal cord injury group and low-intensity exercise group at 28 days after treadmill training(P<0.05).(2)Compared with the control group,the proportion of epidermal growth factor receptor and EdU positive cells was increased in spinal cord injury group(P<0.05).Compared with spinal cord injury group,the proportion of epidermal growth factor receptor and EdU positive cells was increased in low-,moderate-,and high-intensity exercise groups(P<0.05),and the highest was found in moderate-intensity exercise group.Compared with control group,the proportion of glial fibrillary acidic protein positive cells was increased in spinal cord injury group(P<0.05).Compared with spinal cord injury group,the proportion of glial fibrillary acidic protein positive cells was lower in low-,moderate-,and high-intensity exercise groups(P<0.05),and the moderate-intensity exercise group was the lowest.(3)Hematoxylin-eosin staining showed that a large cavity was formed in the injured area of mice with spinal cord injury,and the cavity in the injured area of mice with spinal cord injury decreased after different intensities of treadmill training,and the decrease was most obvious in the moderate-intensity exercise group.(4)These results indicate that low-,moderate-,and high-intensity treadmill training can promote the recovery of motor function of mice with spinal cord injury by activating endogenous neural stem cells,and the effect of moderate-intensity exercise training is the most obvious.

BACKGROUND:We have successfully isolated and cultured neonatal mouse cerebrospinal fluid-contacting neurons in vitro,but there is no study that reports an effective method for isolating and culturing high-purity adult mouse cerebrospinal fluid-contacting neurons.There is no study on whether the self-renewal ability of cerebrospinal fluid-contacting neurons changes with age.OBJECTIVE:To establish a method for isolating and culturing high-purity adult mouse cerebrospinal fluid-contacting neurons in vitro,and to characterize the self-renewal ability of adult mouse cerebrospinal fluid-contacting neurons and neonatal mouse cerebrospinal fluid-contacting neurons in vitro.METHODS:Primary cells containing cerebrospinal fluid-contacting neurons were isolated from the cervical medulla of adult mouse (3 months of age) in adherent culture and transfected with lentivirus fused with multimodal imaging genes.High-purity adult mouse cerebrospinal fluid-contacting neurons were obtained by puromycin screening in suspension culture in complete medium.The expression of neural stem cell markers Nestin and SOX2 was detected by immunofluorescence in adult mouse cerebrospinal fluid-contacting neurons,and the ability of adult mouse cerebrospinal fluid-contacting neurons to form spheres and pass on in vitro was observed.An equal number (5×103/mL) of passage 3 adult mouse and neonatal mouse cerebrospinal fluid-contacting neurons were divided into two groups under the same conditions and inoculated into ultra-low adhesion plates containing complete medium in suspension culture at 5％ CO2,37℃ thermostat,respectively.The self-renewal capacity of adult mouse and neonatal mouse cerebrospinal fluid-contacting neurons was characterized by in vitro spheroid formation,CCK8 assay,qPCR,and western blot assay.RESULTS AND CONCLUSION:(1) High-purity cerebrospinal fluid-contacting neurons were successfully isolated from adult mouse,which expressed Nestin and SOX2 in vitro,and were able to form neurospheres and pass on continuously.(2) The in vitro self-renewal ability of cerebrospinal fluid-contacting neurons in adult mouse was significantly weaker than that of neonatal mouse,and the neurospheres formed by day 4 of cell culture in neonatal mouse were about 150 μm in diameter,whereas the neurospheres formed by adult mouse tactile neurons were only 40 μm in diameter (P<0.0001).(3) CCK8 proliferation assay showed that the proliferative activity of adult mouse cerebrospinal fluid-contacting neurons was significantly weaker than that of neonatal mouse at all time points after culture (P<0.0001).(4) qPCR and western blot assay revealed that the mRNA (P<0.0001) and protein expression levels (P<0.01) of Nestin and SOX2 in cerebrospinal fluid-contacting neurons of adult mouse were significantly decreased compared with those of neonatal mouse.(5) The above results indicated that the self-renewal ability of cerebrospinal fluid-contacting neurons in adult mouse was significantly weaker than that of neonatal mouse in vitro.

Objective To investigate the relationship between the early neutrophil-to-lymphocyte ratio(NLR)in peripheral blood,poor prognosis and early neurological deterioration(END)in patients with acute ischemic stroke(AIS)following intravenous thrombolysis treatment.Methods The retrospective study included 221 AIS patients who underwent intravenous thrombolysis therapy.Demographic information,medical history,clinical and imaging data were collected.Peripheral venous blood samples were drawn before treatment for routine blood tests,and NLR was calculated.The primary endpoint was poor prognosis or death at 3 months after onset,and the secondary endpoint was symptomatic intracranial hemorrhage(SICH)or END within 24 hours after thrombolysis.Logistic regression was used to analyze factors associated with the primary and secondary endpoint events.Receiver operating characteristic(ROC)curve analysis was performed to evaluate the predictive value of NLR for clinical outcomes.Results Logistic regression analysis revealed that female,hypertension,higher baseline NLR(OR=1.968,95%CI:1.516-2.555,P＜0.001)and NIHSS scores were independent risk factors for 3-month poor outcomes in AIS patients.Female,a history of stroke or transient ischemic attack(TIA),higher random blood glucose,elevated baseline NLR(OR=1.317,95%CI:1.028-1.688,P=0.030)and NIHSS scores were independent risk factors for 3-month mortality.Hypertension,elevated homocysteine,higher baseline NLR(OR=1.420,95%CI:1.180-1.709,P＜0.001)and NIHSS scores were independent risk factors for END.There was no significant difference in NLR level between the SICH group(n=5)and the non-SICH group(n=216).ROC curve analysis showed that baseline NLR had high predictive value for 3-month poor outcomes,3-month mortality and END in AIS patients,with AUCs of 0.748(95%CI:0.679-0.817),0.738(95%CI:0.622-0.853)and 0.730(95%CI:0.656-0.804),respectively.There was no predictive value for SICH.Patients in the high NLR group(NLR≥2.63,n=89)had significantly higher baseline NIHSS scores,random blood glucose levels and rates of hypertension,coronary artery disease,END,3-month poor outcomes and 3-month mortality compared to those in the low NLR group(NLR＜2.63,n=132).Conclusion Elevated baseline NLR is associated with the occurrence of END and 3-month poor outcomes in AIS patients following intravenous thrombolysis therapy,suggesting its potential as a biomarker for predicting clinical outcomes in AIS patients.

Purpose To quantitatively compare the diffusion parameters of mono-and biexponential diffusion-weighted imaging models,and to obtain optimal sets of b-values in diffusion-weighted MRI for obtaining monoexponential apparent diffusion coefficient(ADC)close to perfusion-insensitive intravoxel incoherent motion(IVIM)model ADC(ADCIVIM)in identifying of pulmonary solid benign and malignant lesions.Materials and Methods IVIM was performed in 40 patients with solid nodule and masse in Xi'an Gaoxin Hospital from July 2021 to August 2022 using a 3.0T MR imager.Two experienced diagnostic radiologists subjectively evaluated the IVIM images.A single index model was used to calculate ADC values(ADC0-1 000,ADC20-1 000,ADC50-1 000,ADC80-1 000,ADC150-1 000,ADC300-1 000,ADC500-1 000,ADC300,500,1 000,ADC300,800,1 000,ADC300,500,ADC300,800 and ADC300,1 000).The reference standard ADCIVIM value were calculated using a double-exponential model.The physician's measurements between two physicians were measured.The malignant and benign groups were compared and receiver operator characteristic curve for all parameters were analyzed.Results The measurement consistency of ADC values under b value sets and ADCIVIM was very good,and the intraclass correlation coefficient was more significant than 0.75.The differences between ADCIVIM and ADC values in each b group were statistically significant(t=-6.016--2.500,all P<0.05).The area under the curve(AUC)of ADCIVIM was the largest(0.906),with an optimal threshold of 1.271×10-3 mm2/s,a sensitivity of 80.0%and a specificity of 93.0%.The diagnostic efficacy close to ADCIVIM were ADC300,800(AUC=0.891),ADC50-1 000(AUC=0.827)and ADC300,800,1 000(AUC=0.795),respectively.The optimal threshold of ADC300,800 was 1.140×10-3 mm2/s,the sensitivity and specificity were 80.0%and 93.7%,respectively.Conclusion Combining b-values 300 s/mm2 and 800 s/mm2 is recommended as routine scanning parameters for identifying the insensitive monoexponential ADC between benign and malignant solid pulmonary lesions.

BACKGROUND:We have successfully isolated and cultured neonatal mouse cerebrospinal fluid-contacting neurons in vitro,but there is no study that reports an effective method for isolating and culturing high-purity adult mouse cerebrospinal fluid-contacting neurons.There is no study on whether the self-renewal ability of cerebrospinal fluid-contacting neurons changes with age.OBJECTIVE:To establish a method for isolating and culturing high-purity adult mouse cerebrospinal fluid-contacting neurons in vitro,and to characterize the self-renewal ability of adult mouse cerebrospinal fluid-contacting neurons and neonatal mouse cerebrospinal fluid-contacting neurons in vitro.METHODS:Primary cells containing cerebrospinal fluid-contacting neurons were isolated from the cervical medulla of adult mouse (3 months of age) in adherent culture and transfected with lentivirus fused with multimodal imaging genes.High-purity adult mouse cerebrospinal fluid-contacting neurons were obtained by puromycin screening in suspension culture in complete medium.The expression of neural stem cell markers Nestin and SOX2 was detected by immunofluorescence in adult mouse cerebrospinal fluid-contacting neurons,and the ability of adult mouse cerebrospinal fluid-contacting neurons to form spheres and pass on in vitro was observed.An equal number (5×103/mL) of passage 3 adult mouse and neonatal mouse cerebrospinal fluid-contacting neurons were divided into two groups under the same conditions and inoculated into ultra-low adhesion plates containing complete medium in suspension culture at 5％ CO2,37℃ thermostat,respectively.The self-renewal capacity of adult mouse and neonatal mouse cerebrospinal fluid-contacting neurons was characterized by in vitro spheroid formation,CCK8 assay,qPCR,and western blot assay.RESULTS AND CONCLUSION:(1) High-purity cerebrospinal fluid-contacting neurons were successfully isolated from adult mouse,which expressed Nestin and SOX2 in vitro,and were able to form neurospheres and pass on continuously.(2) The in vitro self-renewal ability of cerebrospinal fluid-contacting neurons in adult mouse was significantly weaker than that of neonatal mouse,and the neurospheres formed by day 4 of cell culture in neonatal mouse were about 150 μm in diameter,whereas the neurospheres formed by adult mouse tactile neurons were only 40 μm in diameter (P<0.0001).(3) CCK8 proliferation assay showed that the proliferative activity of adult mouse cerebrospinal fluid-contacting neurons was significantly weaker than that of neonatal mouse at all time points after culture (P<0.0001).(4) qPCR and western blot assay revealed that the mRNA (P<0.0001) and protein expression levels (P<0.01) of Nestin and SOX2 in cerebrospinal fluid-contacting neurons of adult mouse were significantly decreased compared with those of neonatal mouse.(5) The above results indicated that the self-renewal ability of cerebrospinal fluid-contacting neurons in adult mouse was significantly weaker than that of neonatal mouse in vitro.

Purpose To quantitatively compare the diffusion parameters of mono-and biexponential diffusion-weighted imaging models,and to obtain optimal sets of b-values in diffusion-weighted MRI for obtaining monoexponential apparent diffusion coefficient(ADC)close to perfusion-insensitive intravoxel incoherent motion(IVIM)model ADC(ADCIVIM)in identifying of pulmonary solid benign and malignant lesions.Materials and Methods IVIM was performed in 40 patients with solid nodule and masse in Xi'an Gaoxin Hospital from July 2021 to August 2022 using a 3.0T MR imager.Two experienced diagnostic radiologists subjectively evaluated the IVIM images.A single index model was used to calculate ADC values(ADC0-1 000,ADC20-1 000,ADC50-1 000,ADC80-1 000,ADC150-1 000,ADC300-1 000,ADC500-1 000,ADC300,500,1 000,ADC300,800,1 000,ADC300,500,ADC300,800 and ADC300,1 000).The reference standard ADCIVIM value were calculated using a double-exponential model.The physician's measurements between two physicians were measured.The malignant and benign groups were compared and receiver operator characteristic curve for all parameters were analyzed.Results The measurement consistency of ADC values under b value sets and ADCIVIM was very good,and the intraclass correlation coefficient was more significant than 0.75.The differences between ADCIVIM and ADC values in each b group were statistically significant(t=-6.016--2.500,all P<0.05).The area under the curve(AUC)of ADCIVIM was the largest(0.906),with an optimal threshold of 1.271×10-3 mm2/s,a sensitivity of 80.0%and a specificity of 93.0%.The diagnostic efficacy close to ADCIVIM were ADC300,800(AUC=0.891),ADC50-1 000(AUC=0.827)and ADC300,800,1 000(AUC=0.795),respectively.The optimal threshold of ADC300,800 was 1.140×10-3 mm2/s,the sensitivity and specificity were 80.0%and 93.7%,respectively.Conclusion Combining b-values 300 s/mm2 and 800 s/mm2 is recommended as routine scanning parameters for identifying the insensitive monoexponential ADC between benign and malignant solid pulmonary lesions.

Objective To investigate the relationship between the early neutrophil-to-lymphocyte ratio(NLR)in peripheral blood,poor prognosis and early neurological deterioration(END)in patients with acute ischemic stroke(AIS)following intravenous thrombolysis treatment.Methods The retrospective study included 221 AIS patients who underwent intravenous thrombolysis therapy.Demographic information,medical history,clinical and imaging data were collected.Peripheral venous blood samples were drawn before treatment for routine blood tests,and NLR was calculated.The primary endpoint was poor prognosis or death at 3 months after onset,and the secondary endpoint was symptomatic intracranial hemorrhage(SICH)or END within 24 hours after thrombolysis.Logistic regression was used to analyze factors associated with the primary and secondary endpoint events.Receiver operating characteristic(ROC)curve analysis was performed to evaluate the predictive value of NLR for clinical outcomes.Results Logistic regression analysis revealed that female,hypertension,higher baseline NLR(OR=1.968,95%CI:1.516-2.555,P＜0.001)and NIHSS scores were independent risk factors for 3-month poor outcomes in AIS patients.Female,a history of stroke or transient ischemic attack(TIA),higher random blood glucose,elevated baseline NLR(OR=1.317,95%CI:1.028-1.688,P=0.030)and NIHSS scores were independent risk factors for 3-month mortality.Hypertension,elevated homocysteine,higher baseline NLR(OR=1.420,95%CI:1.180-1.709,P＜0.001)and NIHSS scores were independent risk factors for END.There was no significant difference in NLR level between the SICH group(n=5)and the non-SICH group(n=216).ROC curve analysis showed that baseline NLR had high predictive value for 3-month poor outcomes,3-month mortality and END in AIS patients,with AUCs of 0.748(95%CI:0.679-0.817),0.738(95%CI:0.622-0.853)and 0.730(95%CI:0.656-0.804),respectively.There was no predictive value for SICH.Patients in the high NLR group(NLR≥2.63,n=89)had significantly higher baseline NIHSS scores,random blood glucose levels and rates of hypertension,coronary artery disease,END,3-month poor outcomes and 3-month mortality compared to those in the low NLR group(NLR＜2.63,n=132).Conclusion Elevated baseline NLR is associated with the occurrence of END and 3-month poor outcomes in AIS patients following intravenous thrombolysis therapy,suggesting its potential as a biomarker for predicting clinical outcomes in AIS patients.

Objective To construct evaluation indicators for the construction of key specialties in traditional Chinese medicine nursing at the provincial and municipal levels and conduct preliminary verification,providing references for evaluating the level of construction of key specialties in traditional Chinese medicine nursing.Methods From December 2022 to March 2023,based on the Wuli-Shili-Renli system methodology,the content and weight of the evaluation indicators for the construction of key specialties in traditional Chinese medicine nursing at the provincial and municipal levels were determined through literature review,semi-structured interviews,Delphi method,and analytic hierarchy process.Opinion consultation and satisfaction survey were conducted on 46 nursing managers of secondary and above traditional Chinese medicine hospitals in Chongqing in April 2023 to preliminarily verify the application effect of the indicators.Results After 2 rounds of consultation with 16 experts,the questionnaire response rates were 100％and 100％,respectively.The expert authority coefficients were 0.963 and 0.950,respectively,and the coordination coefficients were 0.216～0.307,0.134～0.144,respectively(P＜0.05).Ultimately,an evaluation index for the construction of key specialties in traditional Chinese medicine nursing at the provincial and municipal levels was formed,which includes 3 primary indicators,17 secondary indicators,and 76 tertiary indicators.97.83％nursing managers of hospitals are satisfied with the constructed indicators,believing that the indicator content is clear,comprehensive,professional,and has good practicality.Conclusion The evaluation indicators for the construction of key specialties in traditional Chinese medicine nursing at the provincial and municipal levels are scientific,reliable,which can provide references for the construction of key specialties in traditional Chinese medicine nursing and promote the development of traditional Chinese medicine nursing.

Objective To construct evaluation indicators for the construction of key specialties in traditional Chinese medicine nursing at the provincial and municipal levels and conduct preliminary verification,providing references for evaluating the level of construction of key specialties in traditional Chinese medicine nursing.Methods From December 2022 to March 2023,based on the Wuli-Shili-Renli system methodology,the content and weight of the evaluation indicators for the construction of key specialties in traditional Chinese medicine nursing at the provincial and municipal levels were determined through literature review,semi-structured interviews,Delphi method,and analytic hierarchy process.Opinion consultation and satisfaction survey were conducted on 46 nursing managers of secondary and above traditional Chinese medicine hospitals in Chongqing in April 2023 to preliminarily verify the application effect of the indicators.Results After 2 rounds of consultation with 16 experts,the questionnaire response rates were 100％and 100％,respectively.The expert authority coefficients were 0.963 and 0.950,respectively,and the coordination coefficients were 0.216～0.307,0.134～0.144,respectively(P＜0.05).Ultimately,an evaluation index for the construction of key specialties in traditional Chinese medicine nursing at the provincial and municipal levels was formed,which includes 3 primary indicators,17 secondary indicators,and 76 tertiary indicators.97.83％nursing managers of hospitals are satisfied with the constructed indicators,believing that the indicator content is clear,comprehensive,professional,and has good practicality.Conclusion The evaluation indicators for the construction of key specialties in traditional Chinese medicine nursing at the provincial and municipal levels are scientific,reliable,which can provide references for the construction of key specialties in traditional Chinese medicine nursing and promote the development of traditional Chinese medicine nursing.

BACKGROUND: Ischemic acute kidney injury (AKI) is a common syndrome associated with considerable mortality and healthcare costs. Up to now, the underlying pathogenesis of ischemic AKI remains incompletely understood, and specific strategies for early diagnosis and treatment of ischemic AKI are still lacking. Here, this study aimed to define the transcriptomic landscape of AKI patients through single-cell RNA sequencing (scRNA-seq) analysis in kidneys. METHODS: In this study, scRNA-seq technology was applied to kidneys from two ischemic AKI patients, and three human public scRNA-seq datasets were collected as controls. Differentially expressed genes (DEGs) and cell clusters of kidneys were determined. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis, as well as the ligand-receptor interaction between cells, were performed. We also validated several DEGs expression in kidneys from human ischemic AKI and ischemia/reperfusion (I/R) injury induced AKI mice through immunohistochemistry staining. RESULTS: 15 distinct cell clusters were determined in kidney from subjects of ischemic AKI and control. The injured proximal tubules (PT) displayed a proapoptotic and proinflammatory phenotype. PT cells of ischemic AKI had up-regulation of novel pro-apoptotic genes including USP47 , RASSF4 , EBAG9 , IER3 , SASH1 , SEPTIN7 , and NUB1 , which have not been reported in ischemic AKI previously. Several hub genes were validated in kidneys from human AKI and renal I/R injury mice, respectively. Furthermore, PT highly expressed DEGs enriched in endoplasmic reticulum stress, autophagy, and retinoic acid-inducible gene I (RIG-I) signaling. DEGs overexpressed in other tubular cells were primarily enriched in nucleotide-binding and oligomerization domain (NOD)-like receptor signaling, estrogen signaling, interleukin (IL)-12 signaling, and IL-17 signaling. Overexpressed genes in kidney-resident immune cells including macrophages, natural killer T (NKT) cells, monocytes, and dendritic cells were associated with leukocyte activation, chemotaxis, cell adhesion, and complement activation. In addition, the ligand-receptor interactions analysis revealed prominent communications between macrophages and monocytes with other cells in the process of ischemic AKI. CONCLUSION Together, this study reveals distinct cell-specific transcriptomic atlas of kidney in ischemic AKI patients, altered signaling pathways, and potential cell-cell crosstalk in the development of AKI. These data reveal new insights into the pathogenesis and potential therapeutic strategies in ischemic AKI.
Humans ; Mice ; Animals ; Transcriptome/genetics* ; Ligands ; Kidney/metabolism* ; Acute Kidney Injury/metabolism* ; Ischemia/metabolism* ; Reperfusion Injury/metabolism* ; Sequence Analysis, RNA ; Adaptor Proteins, Signal Transducing/metabolism* ; Tumor Suppressor Proteins/metabolism*