[Current situation and influencing factors of wheezing among children and adolescents aged 3-18 years in 11 cities in China from 2022 to 2023].

Objective: To explore the incidence and influencing factors of wheezing among children and adolescents aged 3-18 years in 11 cities in China from 2022 to 2023. Methods: From October 2022 to August 2023, 11 cities including Xishuangbanna in Yunnan Province, Suqian in Jiangsu Province, Chifeng and Hohhot in Inner Mongolia, Tangshan in Hebei Province, Changzhi in Shanxi Province, Yinchuan in Ningxia Province, Lanzhou and Dingxi in Gansu Province, Linyi in Shandong Province, and Tonghua in Jilin Province were selected as research sites to recruit kindergarten children and primary and secondary school adolescents in local urban areas. A total of 21 959 children and adolescents were included in this study. Demographic information, wheezing data (whether wheezing has occurred in the past and whether wheezing attacks have occurred in the past one year), personal history, family history and other information were collected through questionnaires. The multivariate logistic regression model was used to analyze the influencing factors of wheezing attacks in the past one year. Results: The mean age of 21 959 children and adolescents was (12.09±3.65) years old, and 52.3% (11 480) were boys. The incidence of wheezing history was 3.7% (816 cases), and the incidence of wheezing attacks in the past year was 2.5% (556 cases). The multivariate logistic regression model analysis results showed that compared with older age, girls, full-term natural delivery, no allergic rhinitis, no family history of allergic diseases, no passive smoking, partial diet, natural conception and childbirth, the children aged 3-18 years with young age, male, partial diet, passive smoking, family history of allergic diseases, allergic rhinitis, cesarean section, premature birth, and assisted reproduction had a higher risk of wheezing [OR (95%CI): 0.86(0.84-0.88), 1.27(1.07-1.51), 2.31(1.95-2.75), 2.09(1.76-2.47), 3.5(2.80-4.37), 4.05(3.39-4.83), 1.20(1.02-1.43), 2.26(1.66-3.09), and 1.67(1.01-2.78)]. Conclusion: From 2022 to 2023, the incidence of wheezing among children aged 3-18 years in China is not significantly higher than before, and childhood wheezing may be related to factors such as children's age, gender, dietary habits, family and personal history of allergic diseases, passive smoking, and perinatal period.

[Study on the application value of fecal SDC2 gene methylation detection in colorectal cancer screening of urban residents in Zengcheng District in Guangzhou City].

Objective: To investigate the application value of fecal Syndecan-2 (SDC2) gene methylated SDC2 (mSDC2) detection in colorectal cancer (CRC) screening among urban residents in Guangzhou City. Methods: A cross-sectional study was conducted in Shitan Town, Zengcheng District, Guangzhou City from July to December 2022. A community-based screening program for CRC was conducted among residents aged 40-74 years old. mSDC2 detection was employed in the participants, and those with positive results should be recommended to receive colonoscopy examination. The positive rate of mSDC2 detection, colonoscopy compliance rate, detection rate of intestinal lesions and clinicopathological characteristics were observed. The relationship between cycle threshold (CT) value of mSDC2 and intestinal lesions was explored. Further, the cost-effectiveness of screening was evaluated. Results: A total of 8 189 fecal samples were collected from 8 877 participants with the recovery rate of 92.25%. 8 048 qualified samples were enrolled in this study, consisted of 3 182 males (39.54%) and 4 866 females (60.46%), with the average age of 56 years old (40-74 years). The positive rate of mSDC2 detection was 7.99% (643/8 048), and the compliance rate of colonoscopy was 73.10% (470/643). 20 cases (4.25%) of colorectal cancer, 109 cases (23.19%) of advanced adenoma, 145 cases (30.85%) of non-advanced adenoma, 79 cases (16.81%) of polyps were detected. The detection rate of intestinal lesions was 75.11% and indicated significant differences in gender and age. 20 CRCs included 15 of stage 0-I, 4 of stage Ⅱ-Ⅲ and 1 of unknown stage. The CT value of mSDC2 was negatively correlated with the proportion of advanced colorectal neoplasms (χ2=16.063, P<0.001). The total cost of the screening was 4.339 5 million yuan, the screening benefit was 28.506 2 million yuan, and the benefit-cost ratio was 6.57. Conclusion: The CRC screening strategy of fecal mSDC2 detection combined with colonoscopy has high colonoscopy compliance and detection rate of intestinal lesions, which is conducive to the detection of early CRCs, and has good cost-effectiveness. This study suggests that this method may be applied to the general CRC screening in China and contribute to the prevention of CRC. The CT value of mSDC2 may have a certain suggestion on the malignant degree of intestinal tumors.

Quantitative Intracerebral Iodine Extravasation in Risk Stratification for Intracranial Hemorrhage in Patients with Acute Ischemic Stroke.

BACKGROUND AND PURPOSE: Intracerebral hemorrhage poses a severe threat to the outcomes in patients with postthrombectomy acute stroke. We aimed to compare the absolute intracerebral iodine concentration and normalized iodine concentration ratio in predicting intracerebral hemorrhage in patients postthrombectomy. MATERIALS AND METHODS: Patients with acute anterior circulation large-vessel occlusion who underwent mechanical thrombectomy and had successful recanalization were retrospectively included in the study. Dual-energy CT was performed within 1 hour after mechanical thrombectomy. Postprocessing was performed to measure the absolute intracerebral iodine concentration and the normalized iodine concentration ratio. The correlation between the absolute intracerebral iodine concentration and the normalized iodine concentration ratio was analyzed using the Spearman rank correlation coefficient. We compared the area under the receiver operating characteristic curve of the absolute intracerebral iodine concentration and the normalized iodine concentration ratio using the DeLong test. RESULTS: We included 138 patients with successful recanalization. Of 43 patients who did not have parenchymal contrast staining on postthrombectomy dual-energy CT, 5 (11.6%) developed intracerebral hemorrhage. Among patients (95/138, 68.8%) with parenchymal contrast staining, 37 (38.9%, 37/95) developed intracerebral hemorrhage. The absolute intracerebral iodine concentration was significantly correlated with the normalized iodine concentration ratio (ρ = 0.807; 95% CI, 0.718-0.867; P < .001). The cutoffs of the normalized iodine concentration ratio and absolute intracerebral iodine concentration for identifying patients with intracerebral hemorrhage development were 222.8%, with a sensitivity of 67.6% and specificity of 76.4%, and 2.7 mg I/mL, with a sensitivity of 75.7% and specificity of 65.5%, respectively. No significant difference was found between the areas under the receiver operating characteristic curve for the absolute intracerebral iodine concentration and the normalized iodine concentration ratio (0.753 versus 0.738) (P = .694). CONCLUSIONS: The hemorrhagic transformation predictive power of the normalized iodine concentration ratio is similar to that of the absolute intracerebral iodine concentration in patients with successful recanalization.

[Untargeted metabolomic analysis of serum samples from children with mycoplasma pneumonia in a hospital in Beijing].

Objective: This study was aimed to analyze the untargeted metabolomics of serum samples from children with mycoplasma pneumonia in a hospital in Beijing. Methods: A total of 50 children with mycoplasma pneumonia as the case group were recruited from Department of Pediatrics, China-Japan Friendship Hospital in Beijing from January 2019 to February 2020, and meanwhile 50 age-and gender-matched heathy children were selected and formed the control group. 2 ml venous fasting blood samples was collected from all children. Serum metabolites were quantified by using the untargeted ultra-high performance liquid chromatography/tandem mass spectrometry (UPLC-MS/MS) technique. Unsupervised principle component analysis and (orthogonal) partial least-squares-discriminant analysis were employed to identify differential metabolites between cases and controls. MBRole software was used for pathway enrichment analysis. Results: There were 27 boys and 23 girls in the case group with an average age of (6.0±3.65) years, and the control group consisted of 28 boys and 22 girls with an average age of (6.62±2.64) years. A total of 392 different metabolites were detected. Compared with the control group, 306 metabolites were decreased and 86 increased in case group. Forty-one differential metabolites with variable important in projection (VIP) values larger than 5 and P values less than 0.05 were teased out, and they mainly concentrated on phospholipid. The levels of 38 metabolites were significantly lower in the case group, yet 4 metabolites were significantly higher than that of the control group. Metabolic enrichment analysis showed that different metabolites were related to the biosynthesis of phenylalanine, tyrosine, tryptophan, unsaturated fatty acid, ammonia acyl tRNA and insulin signaling pathway, as well as the metabolism of ABC transporters. Conclusion: The serum untargeted metabolomics differed remarkably between children with mycoplasma pneumonia and healthy children.

[Proliferation of hypertrophic scar fibroblasts inhibited by microRNA-627 targeting IGF-â in hypertrophic scar].

Objective: To investigate the mechanism of microRNA-627(miR-627) inhibiting the proliferation of hypertrophic scar fibroblasts (Fbs) by targeting IGF-I. Methods: The experimental method was used. From October 2019 to January 2020, hypertrophic scar tissues from 6 patients with hypertrophic scar (2 males and 4 females, aged (34±11) years) and the remaining normal skin tissues from 6 patients with trauma (3 males and 3 females, aged (35±13) years) after skin flap transplantation were collected. the above-mentioned 12 patients were admitted to the General Hospital of Northern Theater Command and met inclusion criteria. The mRNA expression of miR-627 was detected by real-time fluorescent quantitative reverse transcription polymerase chain reaction. The 3rd to 5th generations of Fbs were cultured from hypertrophic scar tissue for subsequent experiments. Fbs from hypertrophic scar were divided into miR-627 control group, miR-627 mimic group and miR-627 inhibitor group. The corresponding sequences were transfected respectively. At 0 (immediate), 12, 24, 36 and 48 h after transfection, the cell viability was detected by thiazolyl blue reagent; at 24 h after transfection, the apoptosis was detected by Annexin V-fluorescein-5-isothiocyanate/propidium iodide kit; at 24 h after transfection, the expression levels of IGF-Ⅰ, collagen I and a-SMA were detected by Western blot. The hypertrophic scar Fbs were divided into IGF-Ⅰ wild type + miR-627 control group, IGF- wild type + miR-627 mimics group, IGF-Ⅰ mutant + miR-627 control group. At 48 hours after transfection, the expression of luciferase and renal luciferase were detected by luciferase reporter gene detection kit, and the ratio of the two was calculated to reflect the activity of IGF-Ⅰ. Fbs from hypertrophic scar were divided into miR-627 control group, miR-627 mimic group and miR-627 mimic + IGF-I group, and were transfected with corresponding sequences respectively. At 24 h after transfection, the expression levels of IGF-Ⅰ, type I collagen and a-SMA were detected by Western blot. The number of samples in cell experiment was 3. Analysis of variance, one-way analysis of variance, t test and chi-square test were used to statistic the data. Results: The expression of miR-627 mRNA in hypertrophic scar tissue was 0.47±0.06, which was significantly lower than that in normal tissue 1.12±0.23 (t=15.090, P<0.01). At 12, 24, 36 and 48 hours after transfection, the cell viability of miR-627 mimic group was significantly lower than that of miR-627 control group (t=9.918, 34.370, 13.580, 61.550, P<0.05 or P<0.01); the cell viability of miR-627 inhibitor group was significantly higher than that of miR-627 control group (t=4.722, 8.616, 13.330, 14.000, P<0.05 or P<0.01). At 24 h after transfection, the apoptosis rate of miR-627 mimic group was (10.89±0.35)% significantly higher than that of miR-627 control group (8.42±0.47)% (t=7.301, P<0.01), and that of miR-627 inhibitor group was (5.00±0.22)% significantly lower significantly (t=11.510, P<0.01). At 24 h after transfection, compared with miR-627 control group, miR-627 mimics could significantly down regulate the expression of IGF-Ⅰ, type I collagen and a-SMA (t=25.470, 5.282, 7.415, P<0.05); miR-627 inhibitor could up regulate the expression of IGF-Ⅰ, type I collagen and a-SMA (t=15.930, 8.857, 9.763, P<0.05). At 48 h after transfection, the luciferase/renal luciferase ratio of IGF-Ⅰ in IGF-Ⅰ wild type + miR-627 mimic group was 0.463±0.061, which was significantly lower than that of IGF-Ⅰ wild type + miR-627 control group 0.999±0.011 (t=16.852, P<0.01), The luciferase/renal luciferase ratio of IGF-mutant + miR-627 mimic group was 0.934±0.021, which was similar to that of IGF-Ⅰ mutant+miR-627 control group 0.930±0.023 (t=1.959, P>0.05). After 24 hours of transfection, the protein expressions of IGF-Ⅰ, collagen I and a-SMA in miR-627 mimic group were 1.623±0.070, 1.363±0.042 and 1.617±0.025, which were significantly lower than those in miR-627 control group 2.723±0.045, 2.147±0.067 and 2.533±0.055 (t=22.831, 7.280 and 26.220, P<0.05); The protein expression of miR-627 mimic+IGF-Ⅰ group was 2.477±0.102, 1.760±0.046, 2.387±0.049, which was significantly higher than that of miR-627 mimic group (t=3.83, 8.286, 3.436, P<0.05). Conclusion: miR-627 can inhibit the proliferation of Fbs in hypertrophic scar by targeting IGF-Ⅰ.

[Expression and effect of microRNA-627 in human hypertrophic scar].

Objective: To investigate the expression and effect of microRNA-627 (miR-627) in human hypertrophic scar. Methods: The experimental research method was used. From October 2019 to January 2020, hypertrophic scar tissue from 6 patients with hypertrophic scar (2 males and 4 females, aged (34±11) years) and the remaining normal skin tissue from 6 trauma patients (3 males and 3 females, aged (35±13) years) after flap transplantation were collected. The above-mentioned 12 patients were admitted to the General Hospital of Northern Theater Command and met the inclusion criteria. The mRNA expression of miR-627 was detected by real-time fluorescent quantitative reverse transcription polymerase chain reaction. The 3rd to 5th passages of fibroblasts (Fbs) were isolated from hypertrophic scar tissue and cultured for subsequent experiments after identification. Fbs from hypertrophic scar were divided into miR-627 negative control group, miR-627 mimic group, and miR-627 inhibitor group. The corresponding sequences were transfected respectively. At 0 (immediately), 12, 24, 36, and 48 h after transfection, the cell viability was detected by thiazolyl blue method; at 24 h after transfection, the apoptosis was detected by flow cytometry; at 24 h after transfection, the protein expression levels of insulin-like growth factor Ⅰ (IGF-Ⅰ), type Ⅰ collagen, and α smooth muscle actin (α-SMA) were detected by Western blotting. Two batches of Fbs from hypertrophic scar were used, one batch was divided into IGF-Ⅰ wild type+miR-627 negative control group and IGF-Ⅰ wild type+miR-627 mimic group, and the other batch was divided into IGF-Ⅰ mutant+miR-627 negative control group and IGF-Ⅰ mutant+miR-627 mimic group. The corresponding sequences were transfected respectively. At 48 h after transfection, the expressions of luciferase and renal luciferase were detected by luciferase reporter gene detection kit, and the ratio of the two was calculated to reflect the activity of IGF-Ⅰ. Fbs from hypertrophic scar were divided into miR-627 negative control group, miR-627 mimic alone group, and miR-627 mimic+IGF-Ⅰ group, and were transfected with the corresponding sequences respectively. At 24 h after transfection, the protein expression levels of IGF-Ⅰ, type Ⅰ collagen, and α-SMA were detected by Western blotting. The number of samples in cell experiment was 3. Data were statistically analyzed with analysis of variance for factorial design, one-way analysis of variance, independent sample t test, and chi-square test. Results: The expression of miR-627 mRNA in hypertrophic scar tissue was 0.47±0.06, which was significantly lower than 1.12±0.23 in normal skin tissue (t=15.090, P<0.01). At 12, 24, 36, and 48 hours after transfection, the cell viability of miR-627 mimic group was significantly lower than that of miR-627 negative control group (t=9.918, 34.370, 13.580, 61.550, P<0.05 or P<0.01); the cell viability of miR-627 inhibitor group was significantly higher than that of miR-627 negative control group (t=4.722, 8.616, 13.330, 14.000, P<0.05 or P<0.01). At 24 h after transfection, compared with the apoptosis rate (8.42±0.47)% in miR-627 negative control group, (10.89±0.35)% in miR-627 mimic group was significantly higher (t=7.301, P<0.01), and (5.00±0.22)% in miR-627 inhibitor group was significantly lower (t=11.510, P<0.01). At 24 h after transfection, compared with the cell protein expressions of IGF-Ⅰ, type Ⅰ collagen, and α-SMA in miR-627 negative control group, those in miR-627 mimic group were significantly lower (t=25.470, 5.282, 7.415, P<0.01), and those in miR-627 inhibitor group were significantly higher (t=15.930, 8.857, 9.763, P<0.01). At 48 h after transfection, the luciferase/renal luciferase ratio of IGF-Ⅰ of cells in IGF-Ⅰ wild type+miR-627 mimic group was 0.463±0.061, which was significantly lower than 0.999±0.011 in IGF-Ⅰ wild type+miR-627 negative control group (t=16.852, P<0.01); the luciferase/renal luciferase ratio of IGF-Ⅰ of cells in IGF-Ⅰ mutant+miR-627 mimic group was 0.934±0.021, which was similar to 0.930±0.023 in IGF-Ⅰ mutant+miR-627 negative control group (t=1.959, P>0.05). At 24 h after transfection, the protein expressions of IGF-Ⅰ, type Ⅰ collagen, and α-SMA of cells in miR-627 mimic alone group were 1.623±0.070, 1.363±0.042, and 1.617±0.025, which were significantly lower than 2.723±0.045, 2.147±0.067, and 2.533±0.055 in miR-627 negative control group (t=22.831, 7.280, 26.220, P<0.01); the protein expressions of IGF-Ⅰ, type Ⅰ collagen, and α-SMA of cells in mimic+IGF-Ⅰ group were 2.477±0.102, 1.760±0.046, and 2.387±0.049, which were significantly higher than those of miR-627 mimic alone group (t=3.830, 8.286, 3.436, P<0.05 or P<0.01). Conclusions: miR-627 expression in human hypertrophic scars is down-regulated; miR-627 can inhibit the proliferation and promote the apoptosis of Fbs in human hypertrophic scar by targeted inhibition of IGF-Ⅰ expression.

[Expression and effect of microRNA-205 in human hypertrophic scar].

Objective: To investigate the expression and effect of microRNA-205 (miR-205) in human hypertrophic scar. Methods: The experimental research method was applied. From October 2019 to January 2020, hypertrophic scar tissue from 6 patients with hypertrophic scar (1 male and 5 females, aged (36±7) years) and remaining normal skin tissue from 6 trauma patients (2 males and 4 females, aged (38±9) years) after flap transplantation operation were collected. The above-mentioned 12 patients were admitted to the General Hospital of Northern Theater Command and met the inclusion criteria. Real-time fluorescent quantitative reverse transcription polymerase chain reaction was used to detect the mRNA expressions of miR-205 and thrombospondin-1 (TSP-1). The hypertrophic scar tissue was taken to culture the 3rd to 5th passage of fibroblasts (Fbs) for the follow-up experiments. Two batches of hypertrophic scar Fbs were divided into TSP-1+ miR-205 control group, TSP-1+ miR-205 mimic group, and TSP-1 mutant+ miR-205 control group, TSP-1 mutant+ miR-205 mimic group, which were transfected with the corresponding sequences. At 48 h after transfection, the expressions of luciferase and renal luciferase were detected by luciferase reporter gene detection kit, and the luciferase/renal luciferase ratio was calculated to indicate the activity of TSP-1. Two batches of hypertrophic scar Fbs were collected and divided into miR-205 control group, miR-205 mimic group, and miR-205 inhibitor group and miR-205 control group, miR-205 mimic group, and miR-205 mimic+ TSP-1 group, respectively, which were transfected with the corresponding sequences. At 0 (immediately), 12, 24, 36, and 48 h after transfection, the cell viability was detected by microplate reader. Two batches of hypertrophic scar Fbs were grouped and treated as described in the cell viability detecting experiment. At 24 h after transfection, Hoechst 33258 staining was performed to observe the nuclear shrinkage, so as to reflect the apoptosis of Fbs. The number of samples in cell experiment was three. Data were statistically analyzed with analysis of variance for factorial design, one-way analysis of variance, t test, and chi-square test. Results: The mRNA expression of miR-205 in hypertrophic scar tissue was 0.54±0.05, which was significantly lower than 1.26±0.07 in normal skin tissue (t=8.213, P<0.01). The expression of TSP-1 mRNA in hypertrophic scar tissue was 1.46±0.07, which was significantly higher than 0.68±0.11 in normal skin tissue (t=6.031, P<0.01). At 48 h after transfection, the luciferase/renal luciferase ratio reflecting the TSP-1 activity of cells in TSP-1+ miR-205 mimic group was 0.532±0.028, which was significantly lower than 0.998±0.012 in TSP-1+ miR-205 control group (t=26.500, P<0.01), and the luciferase/renal luciferase ratio of cells in TSP-1 mutant+ miR-205 mimic group was 0.963±0.012, which was close to 0.976±0.010 in TSP-1 mutant+ miR-205 control group (t=0.816, P>0.05). At 12, 24, 36, and 48 h after transfection, the cell viability in miR-205 mimic group was significantly lower than that in miR-205 control group (t=6.169, 12.670, 27.130, 12.670, P<0.05 or P<0.01). At 0, 12, 24, 36, and 48 h after transfection, the cell viability in miR-205 inhibitor group was significantly higher than that in miR-205 control group (t=6.169, 7.221, 7.787, 7.835, 13.030, P<0.05 or P<0.01). At 12, 24, 36, and 48 h after transfection, the cell viability in miR-205 mimic group was significantly lower than that in miR-205 control group and miR-205 mimic+ TSP-1 group (t=8.118, 26.970, 39.550, 42.490, 14.570, 12.240, 36.830, 45.220, P<0.05 or P<0.01). At 24 h after transfection, compared with miR-205 control group, the cell apoptosis in miR-205 mimic group was increased, and the cell apoptosis in miR-205 inhibitor group was decreased. At 24 h after transfection, compared with miR-205 mimic group, the cell apoptosis in miR-205 control group and miR-205 mimic+ TSP-1 group were decreased. Conclusions: miR-205 can inhibit the proliferation and promote the apoptosis of Fbs in human hypertrophic scar by inhibiting the expression of TSP-1, which has the potential to be a therapeutic target for hypertrophic scar.

[Effects and mechanism of eleutheroside E on the growth of human hypertrophic scar fibroblasts].

Objective: To investigate the effects and mechanism of eleutheroside E on the growth of human hypertrophic scar fibroblasts (Fbs). Methods: The experimental research method was used. The hypertrophic scar tissue was collected from 6 patients with hypertrophic scar (1 male and 5 females, aged 20 to 51 (37±8) years) admitted to General Hospital of Northern Theater Command, from October 2018 to March 2019. The third to seventh passages of human hypertrophic scar Fbs were cultured for later experiments. Cells were divided into normal saline group, 100 µmol/L eleutheroside E group, 200 µmol/L eleutheroside E group, and 400 µmol/L eleutheroside E group, and normal saline, eleutheroside E at the final molarity of 100, 200, and 400 µmol/L were added to cells in the corresponding groups. Cells were collected and divided into small interfering RNA (siRNA)-negative control alone group, siRNA-thrombospondin 1 (THBS1) alone group, siRNA-negative control+400 µmol/L eleutheroside E group, and siRNA-THBS1+400 µmol/L eleutheroside E group. Cells in siRNA-negative control alone group and siRNA-negative control+400 µmol/L eleutheroside E group were transfected with siRNA-negative control, cells in siRNA-THBS1 alone group and siRNA-THBS1+400 µmol/L eleutheroside E group were transfected with siRNA-THBS1. At 24 h after transfection, cells in siRNA-negative control alone group and siRNA-THBS1 alone group were added with normal saline, and cells in siRNA-negative control+400 µmol/L eleutheroside E group and siRNA-THBS1+400 µmol/L eleutheroside E group were added with eleutheroside E at the final molarity of 400 µmol/L. At 0 (immediately), 12, 24, 36, and 48 h after treatment, the cell proliferation activity (expressed as absorbance value) was detected by thiazolyl blue assay. Cells were divided into normal saline group, 200 µmol/L eleutheroside E group, 400 µmol/L eleutheroside E group, siRNA-negative control alone group, siRNA-THBS1 alone group, siRNA-negative control+400 µmol/L eleutheroside E group, and siRNA-THBS1+400 µmol/L eleutheroside E group. The corresponding treatments in each group were the same as before. At 24 h after treatment, the apoptosis was observed by Hoechst 33258 staining. Cells were collected and divided into normal saline group, 100 µmol/L eleutheroside E group, 200 µmol/L eleutheroside E group, 400 µmol/L eleutheroside E group, siRNA-negative control alone group, siRNA-THBS1 alone group, siRNA-negative control+400 µmol/L eleutheroside E group, and siRNA-THBS1+400 µmol/L eleutheroside E group. The corresponding treatments in each group were the same as before. At 24 h after treatment, the THBS1 protein level of cells was detected by Western blotting. The number of sample in each group was all 3 at each time point. Data were statistically analyzed with analysis of variance for factorial design, one-way analysis of variance, independent sample t test, and Bonferroni correction. Results: At 0 h after treatment, the absorbance values of cells in normal saline group, 100 µmol/L eleutheroside E group, 200 µmol/L eleutheroside E group, and 400 µmol/L eleutheroside E group were similar (P>0.05). At 12, 24, 36, and 48 h after treatment, the absorbance values of cells in 100 µmol/L eleutheroside E group, 200 µmol/L eleutheroside E group, and 400 µmol/L eleutheroside E group were significantly lower than those of normal saline group (t=7.64, 28.94, 13.69, 5.87, 6.96, 22.83, 14.75, 11.52, 21.09, 20.15, 29.52, 23.12, P<0.05 or P<0.01). At 0 h after treatment, the absorbance values of cells in siRNA-negative control alone group, siRNA-THBS1 alone group, siRNA-negative control+400 µmol/L eleutheroside E group, and siRNA-THBS1+400 µmol/L eleutheroside E group were similar (P>0.05). At 12, 24, 36, and 48 h after treatment, the absorbance values of cells in siRNA-THBS1 alone group and siRNA-negative control+400 µmol/L eleutheroside E group were significantly lower than those in siRNA-negative control alone group (t=7.14, 44.87, 20.67, 40.98, 9.26, 11.08, 15.33, 20.56, P<0.05 or P<0.01); the absorbance values of cells in siRNA-THBS1 alone group, siRNA-negative control+400 µmol/L eleutheroside E group, and siRNA-THBS1+400 µmol/L eleutheroside E group were similar (P>0.05). Compared with that in normal saline group, the numbers of apoptotic cells in 200 µmol/L eleutheroside E group and 400 µmol/L eleutheroside E group were increased at 24 h after treatment. At 24 h after treatment, compared with that in siRNA-negative control alone group, the numbers of apoptotic cells in siRNA-THBS1 alone group and siRNA-negative control+400 µmol/L eleutheroside E group were increased, while the numbers of apoptotic cells in siRNA-THBS1 alone group, siRNA-negative control+400 µmol/L eleutheroside E group, and siRNA-THBS1+400 µmol/L eleutheroside E group were similar. At 24 h after treatment, the protein levels of THBS1 of cells in 100 µmol/L eleutheroside E group, 200 µmol/L eleutheroside E group, and 400 µmol/L eleutheroside E group (0.87±0.12, 0.38±0.07, 0.20±0.09) were significantly lower than 1.83±0.17 in normal saline group (t=16.61, 16.17, 17.29, P<0.01). At 24 h after treatment, the protein levels of THBS1 of cells in siRNA-THBS1 alone group and siRNA-negative control+400 µmol/L eleutheroside E group (0.61±0.07, 0.58±0.07) were significantly lower than 1.86±0.07 in siRNA-negative control alone group (t=71.06, 83.80, P<0.01), and the protein levels of THBS1 of cells siRNA-THBS1 alone group, siRNA-negative control+400 µmol/L eleutheroside E group, and siRNA-THBS1+400 µmol/L eleutheroside E group (0.63±0.11) were similar (P>0.05). Conclusions: Eleutheroside E can inhibit the growth of human hypertrophic scar Fbs by down-regulating the expression of THBS1.

[Expression and effect of microRNA-205 in hypertrophic scar].

Objective: To investigate the expression and effect of microRNA-205 (miR-205) in hypertrophic scar. Methods: The experimental research method were applied. From October 2019 to January 2020, hypertrophic scar tissue from 6 patients with hypertrophic scar [1 male and 5 females, aged (36±7) years], and remaining normal skin tissue from 6 trauma patients [2 males and 4 females, aged (38±9) years] after flap transplantation operation were collected. The above-mentioned 12 patients were admitted to the General Hospital of Northern Theater Command and met the inclusion criteria. Real time fluorescent quantitative polymerase chain reaction was used to detect the mRNA expressions of miR-205 and thrombospondin-1 (TSP-1). The hypertrophic scar tissue was taken to culture the 3rd to 5th passage of fibroblasts (Fbs) for the follow-up experiments. Fbs of hypertrophic scar was divided into TSP-1+miR-205 control group, TSP-1+miR-205 mimic group, TSP-1 mutant+miR-205 control group, TSP-1 mutant +miR-205 mimic group, which were transfected with the corresponding sequences. At 48 h after transfection, the expressions of luciferase and renal luciferase were detected by luciferase reporter gene detection kit, and the luciferase/renal luciferase ratio was calculated to indicate the activity of TSP-1. Two batches of hypertrophic scar Fbs were collected and divided into miR-205 control group, miR-205 mimic group, and miR-205 inhibitor group and miR-205 control group, miR-205 mimic group, and miR-205 mimic+TSP-1 group, respectively, which were transfected with the corresponding sequences. At 0 (immediately), 12, 24, 36, and 48 h after transfection, the cell viability was detected by microplate reader. Two batches of hypertrophic scar Fbs were collected, grouped, and treated as the cell viability detecting experiment. At 24 h after transfection, Hoechst 33258 staining was performed to observe the nuclear shrinkage, so as to reflect the apoptosis of Fbs. The number of samples in cell experiment was 3. Data were statistically analyzed with analysis of variance for factorial design, one-way analysis of variance, and t test. Results: The mRNA expression of miR-205 in hypertrophic scar tissue was 0.54±0.05, which was significantly lower than 1.26±0.07 in normal skin tissue (t=8.213, P<0.01). The expression of TSP-1 mRNA in hypertrophic scar tissue was 1.46±0.07, which was significantly higher than 0.68±0.11 in normal skin tissue (t=6.031, P<0.01). At 48 h after transfection, the luciferase/renal luciferase ratio reflecting the TSP-1 activity of cells in TSP-1+miR-205 mimic group was 0.532±0.028, which was significantly lower than 0.998±0.012 in TSP-1+miR-205 control group (t=26.500, P<0.01), and the luciferase/renal luciferase ratio of cells in TSP-1 mutant+miR-205 mimic group was 0.963±0.012, which was close to 0.976±0.010 in TSP-1 mutant+miR-205 control group (t=0.816, P>0.05). At 12, 24, 36, and 48 h after transfection, the cell viability in miR-205 mimic group was significantly lower than that in miR-205 control group (t=6.169, 12.670, 27.130, 12.670, P<0.05 or P<0.01). At 0, 12, 24, 36, and 48 h after transfection, the cell viability in miR-205 inhibitor group was significantly higher than that in miR-205 control group (t=6.169, 7.221, 7.787, 7.835, 13.030, P<0.05 or P<0.01). At 12, 24, 36, and 48 h after transfection, the cell viability in miR-205 mimic group was significantly lower than that in miR-205 control group and miR-205 mimic+TSP-1 group (t=8.118, 26.970, 39.550, 42.490, 14.570, 12.240, 36.830, 45.220, P<0.05 or P<0.01). At 24 h after transfection, compared with miR-205 control group, the cell apoptosis in miR-205 mimic group was increased, and the cell apoptosis in miR-205 inhibitor group was decreased. At 24 h after transfection, compared with miR-205 mimic group, the cell apoptosis in miR-205 control group miR-205 mimic+TSP-1 group were decreased. Conclusions: miR-205 can inhibit the proliferation and promote the apoptosis of Fbs in hypertrophic scar by inhibiting the expression of TSP-1, which has the potential to be the therapeutic target for hypertrophic scar.

The feasibility of dual-energy CT to predict the probability of symptomatic intracerebral haemorrhage after successful mechanical thrombectomy.

AIM: To study the ability of dual-energy computed tomography (DECT) after successful mechanical thrombectomy (MT) to predict symptomatic intracerebral haemorrhage (sICH) in anterior circulation acute ischaemic stroke (AIS). MATERIALS AND METHODS: From June 2018 to February 2020, 102 AIS patients with DECT performed immediately after successful MT were enrolled prospectively. According to the presence of iodine contrast media extravasation (ICME) on DECT and subsequent sICH development, patients were classified into four groups. The neurological outcome was compared among groups. Imaging parameters, together with clinical factors, were investigated for sICH prediction based on a linear logistic regression model after class-imbalance resolved by Synthetic Minority Sampling Technique (SMOTE) method. RESULTS: Among 102 patients, patients (14.7%, 15/102) with the presence of sICH experienced worse outcomes than others without sICH (p<0.001). No case without ICME was observed with sICH development (0/102). The parameters derived from DECT have excellent performance for sICH prediction after successful MT, which is better than clinical predictive model boosted data (area under the curve [AUC]: DECT 0.87 versus clinical prediction 0.65), cross-validation results (AUC: DECT 0.87 versus clinical prediction 0.65), and original data (AUC: DECT 0.85 versus clinical prediction 0.68). By combining clinical and radiological parameters, the predictive performance for sICH could be further improved with an AUC of 0.90 (95% CI: 0.85-0.96). CONCLUSIONS: Based on DECT parameters acquired immediately after successful MT, the present model was more efficient than the clinical model for accurate prediction of sICH. Rho and ICME volume appeared to be the best parameters for predicting sICH using DECT.

Effect of THBS1 on the Biological Function of Hypertrophic Scar Fibroblasts.

Hypertrophic scarring is a skin collagen disease that can occur following skin damage and is unlikely to heal or subside naturally. Since surgical treatment often worsens scarring, it is important to investigate the pathogenesis and prevention of hypertrophic scarring. Thrombospondin-1 (THBS1) is a matrix glycoprotein that can affect fibrosis by activating TGF-ß1, which plays a key role in wound repair and tissue regeneration; therefore, we investigated the effects of THBS1 on the biological function of hypertrophic scar fibroblasts. THBS1 expression was measured in hypertrophic scars and adjacent tissues as well as normal fibroblasts, normal scar fibroblasts, and hypertrophic scar fibroblasts. In addition, THBS1 was overexpressed or silenced in hypertrophic scar fibroblasts to determine the effects of THBS1 on cell proliferation, apoptosis, and migration, as well as TGF-ß1 expression. Finally, the role of THBS1 in hypertrophic scarring was confirmed in vivo using a mouse model. We found that THBS1 expression was increased in hypertrophic scar tissues and fibroblasts and promoted the growth and migration of hypertrophic scar fibroblasts as well as TGF-ß1 expression. Interestingly, we found that si-THBS1 inhibited the occurrence and development of bleomycin-induced hypertrophic scars in vivo and downregulated TGF-ß1 expression. Together, our findings suggest that THBS1 is abnormally expressed in hypertrophic scars and can induce the growth of hypertrophic scar fibroblasts by regulating TGF-ß1. Consequently, THBS1 could be an ideal target for treating hypertrophic scarring.

Prognosis of Acute Subdural Hematoma in the Elderly: A Systematic Review.

Acute subdural hematoma (aSDH) is among the most common injury types encountered by neurosurgeons, and carries a poor prognosis, particularly in the elderly. As the incidence of aSDH in the elderly population rises, identifying those patients who may benefit from operative intervention is crucial. This systematic review aimed to identify data on prognostic factors or indices, such as the modified frailty index, that may help predict outcome, and hence guide management. A comprehensive search of online databases was conducted by two independent authors, and data on prognostic factors and outcomes were extracted. The quality of the evidence was evaluated using the Grading of Recommendations Assessment, Development and Evaluation (GRADE) criteria. Of 769 studies identified in the initial search, 7 satisfied inclusion and exclusion criteria. Mortality and morbidity varied considerably among studies. Initial Glasgow Coma Scale (GCS) of 3-8 was the most consistently reported negative prognostic feature. Several studies evaluated the impact of medical comorbidities and premorbid frailty, but were limited by small sample size. A previous history of pneumonia was shown to increase the risk of Glasgow Outcome Score (GOS) 1-3 (odds ratio [OR] 6.4 [95% CI 1.6-25.2], p = 0.04) in a single study, which also reported a greater increase in GOS at 3 months in those with fewer than five comorbidities (56% vs. 19%, p < 0.01). There are limited data describing prognostic factors or the use of frailty indices within the specific group of elderly patients with aSDH. Prospective research is needed to evaluate the utility of accurate and validated assessments of frailty to enhance the neurosurgeon's ability to appropriately manage this complex and expanding patient group.

Surface-enhanced Raman spectroscopic analysis of N6-benzylaminopurine residue quantity in sprouts with gold nanoparticles.

A rapid and quantitative method for the determination of N6-Benzylademine (N6-BA) was established through the application of surface-enhanced Raman spectroscopy (SERS). The Raman peak intensities of N6-BA at 1002 cm-1 positively correlated to N6-BA concentrations in sprout extracts. The R2 reached 0.99, and RSDs calculated below 10% at the concentration range of 0.1 ∼5µg mL-1. The average recoveries were 80.0% ∼ 98.2% for blank samples intentionally contaminated at differing levels of 0.04, 0.4, and 1 µg g-1. The whole procedure, including sample preparation and SERS detection, did not exceed 30 min for a set of 6 samples. This study indicates that SERS is a promising technique for rapid tracing analysis and on-site testing of N6-BA.

Spatial Autocorrelation, Source Water and the Distribution of Total and Viable Microbial Abundances within a Crystalline Formation to a Depth of 800 m.

Proposed radioactive waste repositories require long residence times within deep geological settings for which we have little knowledge of local or regional subsurface dynamics that could affect the transport of hazardous species over the period of radioactive decay. Given the role of microbial processes on element speciation and transport, knowledge and understanding of local microbial ecology within geological formations being considered as host formations can aid predictions for long term safety. In this relatively unexplored environment, sampling opportunities are few and opportunistic. We combined the data collected for geochemistry and microbial abundances from multiple sampling opportunities from within a proposed host formation and performed multivariate mixing and mass balance (M3) modeling, spatial analysis and generalized linear modeling to address whether recharge can explain how subsurface communities assemble within fracture water obtained from multiple saturated fractures accessed by boreholes drilled into the crystalline formation underlying the Chalk River Laboratories site (Deep River, ON, Canada). We found that three possible source waters, each of meteoric origin, explained 97% of the samples, these are: modern recharge, recharge from the period of the Laurentide ice sheet retreat (ca. ∼12000 years before present) and a putative saline source assigned as Champlain Sea (also ca. 12000 years before present). The distributed microbial abundances and geochemistry provide a conceptual model of two distinct regions within the subsurface associated with bicarbonate - used as a proxy for modern recharge - and manganese; these regions occur at depths relevant to a proposed repository within the formation. At the scale of sampling, the associated spatial autocorrelation means that abundances linked with geochemistry were not unambiguously discerned, although fine scale Moran's eigenvector map (MEM) coefficients were correlated with the abundance data and suggest the action of localized processes possibly associated with the manganese and sulfate content of the fracture water.

Three-dimensional digitalized virtual planning for saphenous artery flap: a pilot study.

OBJECTIVE: Since the 1970s, research and applications on flap and muscle flap had solved many problems in microsurgical reconstruction. However, the traditional flap design is completely dependent on two-dimensional (2D) images. The purpose of this study was to discuss the methods in the visualization of saphenous artery flap by digitalized technique and its applications by digitalized technique. METHODS: Two adult fresh cadaver specimens, one male and one female, were subject to radiographic computerized tomography (CT) scanning before and after perfused with lead oxide-gelatine mixture, whose collimation are 0.625 mm (120 kV, 110 mA, 512 × 512 matrix). Through Amira 5.4.1 software, the 2D images in DICOM format were transformed into the 3D models of the entire region. The structures of saphenous artery were observed and the digitized visible models of saphenous artery flap were established through three-dimensional (3D) computerized reconstructions methods from these data using Amira 5.4.1 software. Next six cases of soft-tissue defects of the tibia region, involving the exposure bones underwent contrast-enhanced CT angiography of lower limbs utilizing a 64-row multi-slice spiral CT after median cubital vein injection with Ultravist (3.5 ml/s). 2D images from these data in DICOM format were transformed into computer. The structures of saphenous artery flap were observed and measured using Amira 5.4.1 software. Then, all cases were treated by saphenous artery flap. RESULTS: The 3D reconstructed visible models established from these datasets perfectly displayed the saphenous artery flap anatomy. In six cases, the main trunk and branched of the blood vessels in the designed flap were consistent with the surgical findings. The starting point of the saphenous artery to the average distance of the knee clearance were 119.2 ± 9.6 mm, the average diameter of the saphenous artery from the starting point were 1.5 ± 0.3 mm. The range of flap was 8.0 × 5.0 cm to 20.0 × 8.0 cm. All flaps survived well. After 8-24 months' follow-up the knee flexion was 120-140°, the straight 0-10°. There was no case appeared incision infection. CONCLUSIONS: The preoperative use of 3D digitalized virtual planning for the saphenous artery flap improves the surgical accuracy, decreases the operation time and increases the survival rate of the flap.

Sulforaphane induces adipocyte browning and promotes glucose and lipid utilization.

SCOPE: Obesity is closely related to the imbalance of white adipose tissue storing excess calories, and brown adipose tissue dissipating energy to produce heat in mammals. Recent studies revealed that acquisition of brown characteristics by white adipocytes, termed "browning," may positively contribute to cellular bioenergetics and metabolism homeostasis. The goal was to investigate the putative effects of natural antioxidant sulforaphane (1-isothiocyanate-4-methyl-sulfonyl butane; SFN) on browning of white adipocytes. METHODS AND RESULTS: 3T3-L1 mature white adipocytes were treated with SFN for 48 h, and then the mitochondrial content, function, and energy utilization were assessed. SFN was found to induce 3T3-L1 adipocytes browning based on the increased mitochondrial content and activity of respiratory chain enzymes, whereas the mechanism involved the upregulation of nuclear factor E2-related factor 2/sirtuin1/peroxisome proliferator activated receptor gamma coactivator 1 alpha signaling. SFN enhanced uncoupling protein 1 expression, a marker for brown adipocyte, leading to the decrease in cellular ATP. SFN also enhanced glucose uptake and oxidative utilization, lipolysis, and fatty acid oxidation in 3T3-L1 adipocytes. CONCLUSION: SFN-induced browning of white adipocytes enhanced the utilization of cellular fuel, and application of SFN is a promising strategy to combat obesity and obesity-related metabolic disorder.

Alignment of the lower extremity mechanical axis by computer-aided design and application in total knee arthroplasty.

PURPOSE: The success of total knee arthroplasty (TKA) depends on many factors. The position of a prosthesis is vitally important. The purpose of the present study was to evaluate the value of a computer-aided establishing lower extremity mechanical axis in TKA using digital technology. METHODS: A total of 36 cases of patients with TKA were randomly divided into the computer-aided design of navigation template group (NT) and conventional intramedullary positioning group (CIP). Three-dimensional (3D) CT scanning images of the hip, knee, and ankle were obtained in NT group. X-ray images and CT scans were transferred into the 3D reconstruction software. A 3D bone model of the hip, knee, ankle, as well as the modified loading, was reconstructed and saved in a stereolithographic format. In the 3D reconstruction model, the mechanical axis of the lower limb was determined, and the navigational templates produced an accurate model using a rapid prototyping technique. The THA in CIP group was performed according to a routine operation. CT scans were performed postoperatively to evaluate the accuracy of the two TKA methods. RESULTS: The averaged operative time of the NT group procedures was [Formula: see text] min shorter than those of the conventional procedures ([Formula: see text]  min). The coronal femoral angle, coronal tibial angle, posterior tibial slope were [Formula: see text], [Formula: see text], [Formula: see text] in NT group and [Formula: see text], [Formula: see text], [Formula: see text] in CIP group, respectively. Statistically significant group differences were found. CONCLUSIONS: The navigation template produced through mechanical axis of lower extremity may provide a relative accurate and simple method for TKA.

Identification of potential therapeutic targets for melanoma using gene expression analysis.

Metastatic melanoma represents a significant cause of death in patients with melanoma and the frequency is increasing. The aim of this study was to identify potential therapeutic targets for metastatic melanoma. Gene expression profile GSE44660 was downloaded from Gene Expression Omnibus database. A total of 22 samples were analyzed in our study, including 3 specimens of normal melanocytes, 12 specimens of melanoma LNM (lymph node metastasis) and 7 specimens of MBM (melanoma brain metastasis). DEGs (differentially expressed genes) in LNM and MBM were identified respectively using Limma package. GO (Gene Ontology) and KEGG (Kyoto Encyclopedia of Genes and Genomes) pathways analyses of common DEGs between two comparison groups were performed using DAVID, followed by cancer-related genes and transcription factor analysis. PPI (protein-protein interaction) network was constructed by STRING, and significant key genes were selected. Totally, 401 common DEGs were identified. Disease analysis showed that ICAM1 (intercellular adhesion molecule 1) and NBN (nibrin) were related to melanoma. In the PPI network, BIRC5 (baculoviral IAP repeat containing 5), BUB1 (BUB1 mitotic checkpoint serine/threonine kinase), GMNN (geminin, DNA replication inhibitor), AURKA (aurora kinase A), TOP2A (topoisomerase (DNA) II alpha) and BUB1B (BUB1 mitotic checkpoint serine/threonine kinase B) were with higher degree more than 50. ICAM1, NBN, BIRC5, BUB1, BUB1B, GMNN, AURKA and TOP2A may play key roles in the progression and development of melanoma. They may be used as specific therapeutic targets in the treatment of metastatic melanoma. However, further experiments are still needed to confirm our results.