Objective:To explore the predictive value of very low frequency (VLF) for the efficacy of vitamin D treatment in children with postural orthostatic tachycardia syndrome (POTS).Methods:A retrospective case-control study was conducted. A total of 65 POTS children from the Department of Pediatrics of the Second Hospital of Lanzhou University from November 2023 to May 2024 were selected as the research subjects, while 102 healthy children from the same period were selected as the control group. The 25 (OH) D levels between POTS children and healthy children were compared. The frequency of eight main symptoms, including palpitations, headache, cold sweat, blurred vision, chest tightness, dizziness, fainting, and digestive discomfort in the POTS group were analyzed, the symptom scores of the orthostatic intolerance were calculated, and the parameters related to short-range heart rate variability (HRV) during the upright test, head-up tilt test (HUTT) were collected. The correlation between 25 (OH) D levels and symptom scores of orthostatic intolerance symptoms and HRV parameters in children with POTS were analyzed by Pearson correlation analysis. The total symptom scores of children with POTS were compared for 3 months before treatment and 2 months after treatment with vitamin D 800 U/d. The difference in score before and after treatment was ≥2 as responsive group, and the score difference before and after treatment was <2 as non-responsive group. The demographic characteristics including age, gender, height, weight, body mass index, as well as HRV parameters such as the standard deviation of nn intervals (SDNN), the standard deviation of the average NN intervals (SDANN), the root mean square of successive differences (rMSSD), low-frequency (LF), high-frequency (HF), very-low-frequency (VLF) were compared between the responsive group and non-responsive group. Multivariate Logistic regression was analyzed to screen independent influencing factors of vitamin D treatment efficacy for POTS. The marker molecules related to the efficacy of vitamin D treatment in children with POTS were analyzed through receiver operating characteristic (ROC) curve.Results:A total of 167 children were included in this study. There were 65 children in the POTS group, 38 males and 27 females, with the age of (10.6±2.2) years. There were 102 healthy children in the control group, 53 males and 49 females, with the age of (11.2±1.8) years.There was no statistically significant difference in gender and age between the two groups (both P>0.05). The serum 25 (OH) D level in children in the POTS group was (37±18)nmol/L, lower than that in the control group (61±17)nmol/L ( P<0.05).Pearson correlation analysis showed that serum 25(OH)D was positively correlated with VLF, SDNN and HF ( r=0.43, 0.65, 0.36, respectively, all P<0.05). Among the POTS children, there were 48 cases in responsive group and 17 cases in non-responsive group. Univariate analysis revealed that responsive group exhibited significantly lower values in age, SDNN, SDANN, and VLF compared to the non-responsive group (all P<0.05). Multivariate Logistic regression analysis showed that age ( OR=1.47, 95% CI 1.08-2.01), SDNN ( OR=1.01, 95% CI 1.00-1.22) and VLF ( OR=1.01, 95% CI 1.00-1.15) were independent influencing factors in the effect of vitamin D in POTS children (all P<0.05). ROC curve analysis showed that the area under the curve of VLF predicting response to vitamin D treatment for POTS was 0.72 (95% CI 0.58-0.85, P=0.008). When the cutoff value was 1 272 ms 2, the sensitivity and specificity were both 70%. Conclusion:VLF may help predict the efficacy of vitamin D treatment in children with POTS.

Objective:To explore the predictive value of very low frequency (VLF) for the efficacy of vitamin D treatment in children with postural orthostatic tachycardia syndrome (POTS).Methods:A retrospective case-control study was conducted. A total of 65 POTS children from the Department of Pediatrics of the Second Hospital of Lanzhou University from November 2023 to May 2024 were selected as the research subjects, while 102 healthy children from the same period were selected as the control group. The 25 (OH) D levels between POTS children and healthy children were compared. The frequency of eight main symptoms, including palpitations, headache, cold sweat, blurred vision, chest tightness, dizziness, fainting, and digestive discomfort in the POTS group were analyzed, the symptom scores of the orthostatic intolerance were calculated, and the parameters related to short-range heart rate variability (HRV) during the upright test, head-up tilt test (HUTT) were collected. The correlation between 25 (OH) D levels and symptom scores of orthostatic intolerance symptoms and HRV parameters in children with POTS were analyzed by Pearson correlation analysis. The total symptom scores of children with POTS were compared for 3 months before treatment and 2 months after treatment with vitamin D 800 U/d. The difference in score before and after treatment was ≥2 as responsive group, and the score difference before and after treatment was <2 as non-responsive group. The demographic characteristics including age, gender, height, weight, body mass index, as well as HRV parameters such as the standard deviation of nn intervals (SDNN), the standard deviation of the average NN intervals (SDANN), the root mean square of successive differences (rMSSD), low-frequency (LF), high-frequency (HF), very-low-frequency (VLF) were compared between the responsive group and non-responsive group. Multivariate Logistic regression was analyzed to screen independent influencing factors of vitamin D treatment efficacy for POTS. The marker molecules related to the efficacy of vitamin D treatment in children with POTS were analyzed through receiver operating characteristic (ROC) curve.Results:A total of 167 children were included in this study. There were 65 children in the POTS group, 38 males and 27 females, with the age of (10.6±2.2) years. There were 102 healthy children in the control group, 53 males and 49 females, with the age of (11.2±1.8) years.There was no statistically significant difference in gender and age between the two groups (both P>0.05). The serum 25 (OH) D level in children in the POTS group was (37±18)nmol/L, lower than that in the control group (61±17)nmol/L ( P<0.05).Pearson correlation analysis showed that serum 25(OH)D was positively correlated with VLF, SDNN and HF ( r=0.43, 0.65, 0.36, respectively, all P<0.05). Among the POTS children, there were 48 cases in responsive group and 17 cases in non-responsive group. Univariate analysis revealed that responsive group exhibited significantly lower values in age, SDNN, SDANN, and VLF compared to the non-responsive group (all P<0.05). Multivariate Logistic regression analysis showed that age ( OR=1.47, 95% CI 1.08-2.01), SDNN ( OR=1.01, 95% CI 1.00-1.22) and VLF ( OR=1.01, 95% CI 1.00-1.15) were independent influencing factors in the effect of vitamin D in POTS children (all P<0.05). ROC curve analysis showed that the area under the curve of VLF predicting response to vitamin D treatment for POTS was 0.72 (95% CI 0.58-0.85, P=0.008). When the cutoff value was 1 272 ms 2, the sensitivity and specificity were both 70%. Conclusion:VLF may help predict the efficacy of vitamin D treatment in children with POTS.

Objective:To investigate the effect of activation of microglia in prefrontal cortex on long-term spatial memory in post-stroke depression mice.Methods:Forty-eight male C57BL/6 mice were divided into sham operation group, stroke group, post-stroke depression group and depression group according to the random number table method with 12 in each group, and 36 mice were divided into solvent group, enrofloxacin group and minocycline group according to the random number table method with 12 in each group.Middle cerebral artery occlusion (MCAO) was use to establish the stroke model, and forced swimming was used to establish the depression model.The post-stroke depression model mice were received MCAO first and then received forced swimming on the 4th day after stroke to establish the model.Mice in enrofloxacin group and minocycline group were treated with enrofloxacin and minocycline injection once a clay for 14 days from the 5th day after stroke, respectively.Forced swimming test and sugar water preference test were used to evaluate the depression of mice in each group, Morris water maze test was used to detect the spatial memory function of mice in each group, and Nissl staining and immunofluorescence staining were used to detect the neuronal function and the number and type of microglia activation.The expression of inflammatory cytokines IL-6 and IL-1β were detected by Western blot.GraphPad Prism 8.0.1 statistical software was used for statistical analysis.The single factor variance analysis was used to compare the difference among multiple groups, and pairwise comparison was performed with SNK- q test. Results:(1) There were statistically significant differences in depression, learning and memory, neuron damage, activation of microglia, inflammatory factors and other indicators in sham operation group, stroke group, post-stroke depression group and depression group ( F=43.58-255.70, all P<0.05). Compared with stroke group, post-stroke depression group had longer floating immobility time ((222.70±29.12) s, (79.25±46.78) s, P<0.05), the preference rate of sugar water was significantly lower ( (49.44±6.19) %, (84.49±4.73) %, P<0.05), and the average value of platform approach after correction was higher((125.00±9.95) mm, (96.79±12.57) mm, P<0.05), Nissl bodies expression was lower ((53.50±15.78) cells /mm 2, (85.67±17.52) cells /mm 2, P<0.05), NeuN positive expression rate was lower ((29.78±3.70) %, (45.73±4.51) %, P<0.05), the percent of M1 microglia expression was significantly higher ((75.55±8.84) %, (58.19±5.69) %, P<0.05), the percent of M2 microglia expression was lower ((43.46±5.11)%, (57.14±5.40)%, P<0.05), and the expression levels of IL-6 ((1.14±0.03), (0.94±0.05), P<0.05) and IL-1β((1.17±0.03), (0.56±0.04), P<0.05) were significantly higher.(2) Depression, learning and memory, neuron injury, activation of microglia, inflammatory factors and other indicators of mice in solvent group, enrofloxacin group and minocycline group were significantly different ( F=7.13-94.35, all P<0.05). Compared with enrofloxacin group, mice in minocycline group had shorter floating immobility time ((169.30±13.04) s, (224.30±22.60) s, P<0.05) and higher sugar water preference rate ((62.81±7.75) %, (47.71±8.11) %, P<0.05), the mean value of platform approach estimation after water maze correction was lower ((97.66±14.56) mm, (120.20±12.08) mm, P<0.05), and the expression level of Nissl bodies was higher ((80.17±10.55) cells /mm 2, (52.00±8.94) cells /mm 2, P<0.05), NeuN expression rate was high ((45.04±3.62) %, (28.88±4.50) %, P<0.05), Iba-1 expression was lower ((97.33±10.67) cells/mm 2, (112.50±6.54)cells/mm 2, P<0.05), the percent of M1 microglia expression was lower ((54.43±5.22) %, (73.82±6.88) %, P<0.05), and the percent of M2 microglia expression was significantly higher ((51.86±6.22) %, (36.30±5.72) %, P<0.05). The expression levels of IL-6 ((0.75±0.06), (1.21±0.07), P<0.05) and IL-1β ((0.61±0.06) (1.09±0.09), P<0.05) were lower. Conclusion:The long-term spatial memory impairment of post-stroke depression mice is aggravated, which is related to the neuron damage caused by increased activation of M1 microglia in PFC area.Inhibition of M1 microglia by minocycline can effectively improve the spatial memory ability of mice.

Objective:To investigate the effect of Spautin-1 (an inhibitor of autophagy) on improving anxiety-like behaviors and its mechanism in mice after traumatic brain injury (TBI).Methods:Thirty-six C57BL/6 mice were randomly divided into sham-operated group, TBI group, and TBI+Spautin-1 group ( n=12); TBI models in the latter two groups were established by modified Feeney free fall epidural impingement method. Mice in TBI+Spautin-1 group were administered with Spautin-1 (2 μL, 10 mmol/L) into the lateral ventricle 10 min after modeling, but mice in the other two groups were only injected with same volume of solvent. Neuropathy Symptom Score (NSS) was used to evaluate the functions of motor, sensory and reflexes of mice on 1 st, 7 th and 14 th d of modeling. On 15 th and 16 th d of modeling, open field test and elevated plus maze test were used to evaluate the anxiety-like behaviors in mice. The number of Nissl bodies in the amygdala of mice was calculated by Nissl staining 16 d after modeling. The numbers of neuron specific nucleoprotein (NeuN) positive cells, interleukin (IL)-18 and IL-1β positive astrocytes in the amygdala were detected by immunofluorescent staining. Western blotting was used to detect the autophagy-and pyrotopic-associated protein expressions in the amygdala region of mice. Results:(1) As compared with the sham-operated group, the TBI group and TBI+Spautin-1 group had significantly increased NSS scores on 1 st and 7 th d of modeling ( P<0.05). (2) Open field test showed that as compared with the sham-operated group, the TBI group and TBI+Spautin-1 group had significantly smaller number of crossing grids, significantly decreased percentage of time spending in the central zone ([central area residence time/total time] × 100%), significantly decreased percentage of frequencies entering into opening arm (OE) (OE/[OE+frequencies of entering closing arm]×100%) and opening arm time (OT) percentage (OT/[OT+time of closing arm]×100%); as compared with the TBI group, TBI+Spautin-1 group had significantly larger number of crossing grids, and significantly increased time percentage spending in the central zone, OE percentage, and OT percentage ( P<0.05). (3) As compared with sham-operated group, the TBI group and TBI+Spautin-1 group had significantly smaller numbers of Nissl bodies and NeuN positive cells in the amygdala of mice ( P<0.05); as compared with TBI group, TBI+Spautin-1 group had significantly larger numbers of Nissl bodies and NeuN positive cells in the amygdala of mice ( P<0.05). (4) As compared with the sham-operated group, the TBI group and TBI+Spautin-1 group had significantly increased percentages of IL-1β and IL-18 positive astrocytes in amygdala of mice ( P<0.05); as compared with the TBI group, the TBI+Spautin-1 group had significantly decreased percentages of IL-1β and IL-18 positive astrocytes in amygdala of mice ( P<0.05). (5) As compared with sham-operated group, the TBI group and TBI+Spautin-1 group had significantly higher protein expressions of NOD-like receptor family protein 3 (NLRP3), activated cysteine aspartate protease-1 (Caspase-1), pore-forming protein D-N terminal fragment (GSDMD-N), ubiquitin specific peptidase (USP) 13 and B-lymphocytoma-2 interacting protein (Beclin1), and statistically higher ratio of microtubule-associated protein 1 light chain (LC)3 II/LC3 I ( P<0.05); as compared with TBI group, the TBI+Spautin-1 group had significantly decreased protein expressions of NLRP3, activated Caspase-1, GSDMD-N, USP13 and Beclin1 in the amygdala, and statistically lower ratio of LC3 II/LC3 I ( P<0.05). Conclusion:Spautin-1 improves the anxiety-like behaviors in mice after TBI, whose mechanism may be associated with the inhibition of astrocytic pyroptosis in the amygdala.

Objective::Older patients with comorbidity, such as coronary heart disease (CHD) and malignant gastrointestinal tumors, are at a high risk of bleeding events. However, risk prediction models based on risk factor assessment remain unclear. This study aimed to establish an individualized bleeding risk assessment system based on the analysis of 10-year inpatient clinical big data.Methods::Total clinical data of 56,819 patients with CHD and 25,988 patients with malignant digestive tract tumors (admitted from January 2008 to December 2017) were retrospectively collected at the First and Second Medical Centers of Chinese People’s Liberation Army General Hospital. Among them, 1307 patients with CHD and malignant digestive tract tumors were screened as the derivation cohort. The dependent variable was the occurrence of major clinical bleeding events. Baseline statistics and hypothesis tests of differences were performed for independent variables according to the occurrence of bleeding. Decision Tree, eXtreme Gradient Boosting (XGBoost), logistic regression, and random forest models were used for comparison. The accuracy, sensitivity, specificity, and area under the receiver operating characteristic curve (AUC-ROC) were applied as the criteria for evaluating and verifying model performance. To evaluate this developed model, another cohort comprising 454 patients (admitted from January 2018 to December 2019) was prospectively enrolled as the validation cohort based on the same inclusion and exclusion criteria.Results::Among the 64 variables with <50% missing values, the recursive feature elimination method with a random forest model was used to screen the selected variables. The highest accuracy was obtained following the selection of 10 scalars, and the final model was constructed accordingly. XGBoost demonstrated the best performance comprehensively. The AUC-ROC of this model was 0.981, with an accuracy, sensitivity, and specificity of 0.939, 0.950, and 0.927, respectively. In the validation cohort, the AUC-ROC, accuracy, sensitivity, and specificity of the XGBoost model were 0.702, 0.718, 0.636, and 0.725, respectively. The rate of major bleeding events has a positive correlation with the bleeding risk score quintiles. To allow for convenient clinical application, a smartphone application was developed for easy access and calculation (http://fir.master-wx.com/sghr).Conclusion::We successfully established a risk model and score for predicting bleeding events in older patients with comorbidity, such as CHD and gastrointestinal cancer.

Objective::Older patients with comorbidity, such as coronary heart disease (CHD) and malignant gastrointestinal tumors, are at a high risk of bleeding events. However, risk prediction models based on risk factor assessment remain unclear. This study aimed to establish an individualized bleeding risk assessment system based on the analysis of 10-year inpatient clinical big data.Methods::Total clinical data of 56,819 patients with CHD and 25,988 patients with malignant digestive tract tumors (admitted from January 2008 to December 2017) were retrospectively collected at the First and Second Medical Centers of Chinese People’s Liberation Army General Hospital. Among them, 1307 patients with CHD and malignant digestive tract tumors were screened as the derivation cohort. The dependent variable was the occurrence of major clinical bleeding events. Baseline statistics and hypothesis tests of differences were performed for independent variables according to the occurrence of bleeding. Decision Tree, eXtreme Gradient Boosting (XGBoost), logistic regression, and random forest models were used for comparison. The accuracy, sensitivity, specificity, and area under the receiver operating characteristic curve (AUC-ROC) were applied as the criteria for evaluating and verifying model performance. To evaluate this developed model, another cohort comprising 454 patients (admitted from January 2018 to December 2019) was prospectively enrolled as the validation cohort based on the same inclusion and exclusion criteria.Results::Among the 64 variables with <50% missing values, the recursive feature elimination method with a random forest model was used to screen the selected variables. The highest accuracy was obtained following the selection of 10 scalars, and the final model was constructed accordingly. XGBoost demonstrated the best performance comprehensively. The AUC-ROC of this model was 0.981, with an accuracy, sensitivity, and specificity of 0.939, 0.950, and 0.927, respectively. In the validation cohort, the AUC-ROC, accuracy, sensitivity, and specificity of the XGBoost model were 0.702, 0.718, 0.636, and 0.725, respectively. The rate of major bleeding events has a positive correlation with the bleeding risk score quintiles. To allow for convenient clinical application, a smartphone application was developed for easy access and calculation (http://fir.master-wx.com/sghr).Conclusion::We successfully established a risk model and score for predicting bleeding events in older patients with comorbidity, such as CHD and gastrointestinal cancer.

Objective: To investigate the effect of mesenchymal stem cells (MSCs) on apoptosis of breast cancer cell line MCF-7 induced by cisplatin (DDP), MSCs derived from breast cancer (BC-MSCs) or adjacent non-cancerous tissues (BN-MSCs) were isolated, cultured and identified. Methods: BC-MSCs and BN-MSCs were isolated and cultured by tissue adherent method. The differentiation potential of BC-MSCs was detected by osteogenic and adipogenic induction, and cell surface markers of BC-MSCs and BN-MSCs were evaluated by flow cytometry. MCF-7 cells were co-treated with DDP and conditioned medium (CM) collected from BC-MSCs and BN-MSCs after being cultured for 48 hours, respectively. Inhibition rate of cell proliferation was evaluated by MTT. Cell apoptosis and viability were detected by MUSE cell analyzer. Cytokines in MSC-CM were detected by Luminex liquid chip. Interleukin 6 (IL-6) mRNA expressions in MCF-7 cells with different treatment were detected by RT-PCR. Results: The morphology of BC-MSCs and BN-MSCs successfully isolated and cultured was uniform fibroblast-like clusters under the microscope. These cells expressed high levels of CD29 and CD44, but neither CD14 nor CD34 were detected. MSCs could also differentiate into osteoblasts and adipocytes after specific induction. After treatment with 2.5, 5, 10, 20, 40 and 80 μmol/L DDP, the inhibitory rates of proliferation of MCF-7 cells in DDP group were (17.33±2.00)%, (22.37±0.73)%, (30.77±1.23)%, (44.93±1.27)%, (62.03 ±1.97)% and (73.93±1.10)%, respectively. While the inhibitory rates of DDP+ BC-MSCs group were (8.27±0.63)%, (11.50±1.30)%, (20.57±0.93)%, (32.60 ±1.90)%, (52.27±0.73)% and (62.13±2.17)%, respectively. The inhibitory rates of DDP+ BN-MSCs group were (12.90±1.60)%, (16.53±2.87)%, (25.90±1.50)%, (39.40±2.40)%, (57.40±0.70)% and (69.03±1.07)%, respectively. The inhibitory rates of DDP+ BC-MSCs group were significantly lower than those of DDP group (P<0.05). The apoptotic rates of MCF-7 cells in DDP group, DDP+ BC-MSCs group and DDP+ BN-MSCs group were (47.77±1.98)%, (29.20±2.12)% and (37.92±2.21)%, respectively. The apoptotic rates of DDP group was significantly higher than that of DDP+ BC-MSCs group (P<0.05). The cell viabilities of MCF-7 in DDP group, DDP+ BC-MSCs group and DDP+ BN-MSCs group were 0.52±0.02, 0.72±0.02 and 0.64±0.02, respectively. The cell viability of DDP group was significantly lower than that of DDP+ BC-MSCs group (P<0.05). The result of Luminex liquid chip analysis showed that, the level of IL-6 in BC-MSCs group increased 2.50±0.68 fold when compared with BN-MSCs group (P<0.05). The relative expressions of IL-6 mRNA in DDP group and DDP+ BC-MSCs group were 1.02±0.10 and 7.58±0.55, respectively, with a statistically significant difference (P<0.01). The apoptotic rates of MCF-7 cells in DDP+ BC-MSCs group with or without IL-6 neutralizing antibody were (27.41±1.95)% and (42.45±2.87)%, respectively, with a statistically significant difference (P<0.05). The cell viabilities of MCF-7 cells in DDP+ BC-MSCs group with or without IL-6 neutralizing antibody were (72.40±2.60)% and (59.76±3.89)%, respectively, with a statistically significant difference (P<0.05). Conclusions BC-MSCs and BN-MSCs have been isolated and cultured successfully. Compared with BN-MSCs, BC-MSCs could attenuate the effect of DDP on MCF-7 cells, evidently decrease the apoptosis and increase the proliferation and vitality in an IL-6 dependent manner.

Objective To evaluate the effects of early rehabilitation on development of infants with cerebral injury in pefinatal period.Methods 84 high risk infants with cerebral injury in pefinatal period were divided into two groups.Infants in the control group were treated with conventional medicament and those in intervention group received physical,visual,aural and haptic stimuli combined with Vojta and Bobath approach in addition.Clinical data and Gesell Development Schedules of 1 year old were used to evaluate the effect.Results The incidence of improvement was 92.86% in the intervention group and 73.81% in the control group(P<0.05).The development quotient of Gesell Development Schedules were obviously higher in the intervention group than in the control group in all the functional areas(P<0.05).Conclusion The early rehabilitation can improve the cerebral development in high risk infants with cerebral injury in pefinatal period.