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Objective: To establish a convenient non-invasive tracheal perfusion method for constructing a mouse model of silicosis-induced pulmonary fibrosis. Methods: The specific pathogen-free C57BL/6 mice were randomly divided into control group and model group, with 15 mice in each group. After anesthesia, a 22G arteriovenous indwelling needle was used to inset into the trachea through the mice's mouth. The model group mice were perfused with 0.1 mL of silica suspension with a mass concentration of 25 g/L, and the mice in the control group were perfused with an equal volume of 0.9% sodium chloride solution. On the 7th, 14th, and 30th day after modeling, the body weight of the mice was measured, and the lung tissue morphology and pathological changes were observed. The expression of α-smooth muscle actin (α-SMA) and collagen type 1 alpha 1 (COL1A1) protein in lung tissue of mice was detected by immunofluorescence on the 30th day after modeling. Results: There was no death of mice in the two groups during the experiment. There was no significant difference in body weight between the two groups (P>0.05). The lung tissues of the mice in the model group were pinkish-gray and uneven in color on the 7th and 14th days after dust exposure. On the 30th day after dust exposure, the lung tissue of the mice in the model group was gray and hard, and unevenly distributed silicon nodules were visible by the naked eyes. The histopathology results of lung tissue showed that compared with the mice in control group, the model group mice exhibited persistent aggravation of pulmonary inflammation, thickening of alveolar septum, infiltration of inflammatory cells gradually clustering into clumps, and an increasing number of fibrous foci.On the 30th day after dust exposure, the relative expression of α-SMA and COL1A1 proteins in the lung of the model group was higher than those in the control group (median： 72.59 vs 5.91, 35.62 vs 10.07, both P<0.05). Conclusion: The method of tracheal perfusion silica suspension of mice using 22G arteriovenous indwelling needle can successfully construct an animal model of silicosis fibrosis. This method is convenient, safe and effective, and is worth promoting.
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Objective @#To examine the diagnostic and prognostic value of long non-coding RNA (lncRNA) JPX in mesothelioma, so as to provide insights into diagnosis and prognosis of mesothelioma. @* Methods@# Patients with clinically definitive diagnosis of mesothelioma from 2015 to 2019 that were sampled from asbestos processing plants in Zhejiang Province from 2015 to 2019 were recruited in the mesothelioma group, while healthy residents without asbestos exposure or asbestos-related diseases in the same area served as controls. Participants' demographics, pathologic diagnosis and imaging features were collected, and the expression of blood lncRNA JPX was detected using lncRNA microarrays. The diagnostic value of lncRNA JPX for mesothelioma was evaluated using the receiver operating characteristic (ROC) curve, and the correlation between lncRNA JPX expression and prognosis was examined among mesothelioma patients using survival analysis. @* Results@# There were 17 subjects in the mesothelioma group, with a mean age of (65.71±8.36) years, and 34 subjects in the controls, with a mean age of (64.24±8.70) years. LncRNA microarray detected significantly high lncRNA JPX expression in mesothelioma patients, and higher blood lncRNA JPX expression was detected in the mesothelioma group than in the control group [median (interquartile range), 1.10 (1.31) vs. 0.89 (0.54); t'=-2.300, P=0.034]. The area under the ROC curve was 0.673 (95%CI: 0.507-0.839, P=0.046), and if the cutoff was 1.759, the sensitivity and specificity were 35.3% and 100.0%, respectively. Survival analysis showed no significant difference in the survival rate of mesothelioma patients between the high lncRNA JPX expression group and the low expression group (χ2=0.212, P=0.645). @*Conclusions@# LncRNA JPX overexpression is detected in the blood of patients with mesothelioma, and lncRNA JPX expression presents a diagnostic value for mesothelioma; however, it shows little prognostic value for mesothelioma.
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Objective@#To investigate the effect of chrysotile exposure on ribosomal DNA (rDNA) copy number and DNA damage response, so as to provide insights into the mechanism of asbestos-induced carcinogenesis. @*Methods@#Human pleural mesothelial MeT-5A cells were treated with chrysotile suspensions at doses of 1.25, 2.5 and 5 μg/cm2 (low-, medium-, high-dose group), while PBS served as controls. MeT-5A cells were harvested 6, 24, 48 and 72 h post-treatment, and the rDNA copy numbers and the BIRC5, HRAS, GINS4 and RRM2 mRNA expression were determined using a quantitative real-time PCR (qPCR) assay. The apoptosis of MeT-5A cells and DNA damage were detected using Muse cell analyzer. The rDNA copy numbers, DNA damage responses and BIRC5, HRAS, GINS4 and RRM2 mRNA expression were compared in MeT-5A cells treated with different doses of chrysotile suspensions.@*Results@#There were significant differences in 45S rDNA copy numbers among low-, medium-, high-dose groups and the control groups 6, 48 and 72 h post-treatment with chrysotile suspensions, and significantly lower 45S rDNA copy numbers were measured in low-, medium- and high-dose groups than in the control group 6 h post-treatment, while significantly higher 45S rDNA copy numbers were found in the high-dose group than in low- and medium-dose groups 48 and 72 h post-treatment (all P<0.05). There were significant differences in 5S rDNA copy numbers among low-, medium-, high-dose groups and the control groups 24, 48 and 72 h post-treatment with chrysotile suspensions, and significantly lower 5S rDNA copy numbers were measured in medium- and high-dose groups than in the control group 24 and 48 h post-treatment, while significantly lower 5S rDNA copy numbers were found in medium- and high-dose groups than in the low-dose group 24, 72 h post-treatment (all P<0.05). There were significant differences in the overall apoptotic rate of MeT-5A cells among groups at different time points, and the overall apoptotic rate of MeT-5A cells were significantly higher in medium- and high-dose groups than in the control group (all P<0.05), with late-stage apoptosis predominantly detected. There were significant differences in the rates of ATM activation and DNA double-strand break in MeT-5A cells among groups 72 h post-treatment, and higher rates of ATM activation and DNA double-strand break were measured in medium- and high-dose groups than in the control group (all P<0.05). In addition, there were significant differences in the relative mRNA expression of BIRC5, HRAS, GINS4 and RRM2 genes among groups 24 and 48 h post-treatment, and significantly lower BIRC5, HRAS, GINS4 and RRM2 mRNA expression was quantified in medium- and high-dose groups than in the control group (all P<0.05).@*Conclusion@#Exposure to chrysotile may induce rDNA copy number variations and altered expression of nucleolar proteins in human pleural mesothelial cells, which may be involved in the regulation of DNA damage responses.
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Objective@#To explore sleep duration and associated factors among children aged 3-6 years old in Zhejiang Province.@*Methods@#A total of 7 034 children aged 3-6 years from 24 kindergartens in Zhejiang province were selected through clustered sampling method. Through parental questionnaire, child sleep duration and associated factors were colleted. Possible influencing factors of sleep insufficiency were analyzed by logistic regression model.@*Results@#The rate of insufficient sleep was 56.1%, with 57.1% for boys and 55.1% for girls in Zhejiang Province. The rate of insufficient sleep increased with the increase of age, which was 46.7%, 49.8%, 50.2%, 55.2%, 64.0%, 65.8% and 68.1% for 3.0, 3.5, 4.0, 4.5, 5.0, 5.5 and 6.0 years old group (χ 2=45.71, P<0.01), respectively. The rate of insufficient sleep was 59.9%, 54.7% and 45.6% for children with <1 h/d, 1-2 h/d and ≥2 h/d outdoor physical activity (χ 2=67.10, P<0.01). Multivariate Logistic regression model showed that children older than 4.5 years old, whose caregiver was other than parents, outdoor activity time< 1 h/d and 1-2 h/d, maternal education below junior middle school and senior high school were positiviely associated with sleep insufficiency [OR(95%CI) was 1.29(1.07-1.54), 1.97(1.64-2.37), 2.04(1.69-2.46), 2.35(1.82-3.03), 1.21(1.08-1.35), 1.73(1.49-2.01), 1.47(1.26-1.72), 1.35(1.15-1.59), 1.16(1.01-1.32), P<0.05].@*Conclusion@#Sleep insufficiency among preschool children is associated with multiple factors, increase of age, insufficient outdoor activity and overdose screen time in particular. The finding calls for outdoor physical activity promotion among preschool children, especially for those whose caregivers are those other than parents.
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Based on the current situation of pediatric surgery practice teaching in Qilu Hospital of Shandong University, 10 mind maps of common clinical diseases made by Imindmap 11 were applied to the practice teaching and teaching assessment through WeChat public platform. Mind map not only revealed the relationship between related disciplines and reappeared the clinical thinking process, but also greatly improved the enthusiasm and efficiency of interns' learning, and trained their innovative spirit and practical ability. At the same time, as a formative evaluation method, mind map facilitates teachers to evaluate and improves teaching activities. What’s more, it effectively improves the quality of teaching, and is worth promoting in clinical teaching.
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Objective@#To investigate whether the cells with the characteristic of mesenchymal stem cells (MSCs) could be found in rat cerebral cortex.@*Methods@#Rat cerebral cortex samples were digested with typeⅡ collagenase into single cells. Culture medium for MSCs was used for cell culture. The morphology of the cells was observed under a microscope. MSC-specific surface markers were detected by flow cytometry. Differentiation potential favoring adipogenesis or osteogenesis of the subcultured cells was analyzed. Lymphocyte transformation test (LTT) was performed to analyze their immunosuppressive function. Bone marrow MSCs (BM-MSCs) were used as positive control cells in all experiments.@*Results@#The fibroblast-like cells isolated from the cerebral cortex of rats were morphologically heomeogeneous after cell culture and grew in a spiral pattern. On the surface of these cells, CD29, CD90 and CD146 were highly expressed at mRNA level, while the expression of CD45 and CD31 at mRNA level were not detected. Moreover, they could be induced to differentiate into adipocytes and osteoblasts. Results of LTT showed that they were functionally equivalent to BM-MSCs in inhibiting the proliferation of spleen lymphocytes induced by concanavalin A.@*Conclusions@#This study suggested that the cells with the characteristic of MSCs could be detected in rat cerebral cortex.
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Objective To investigate whether the cells with the characteristic of mesenchymal stem cells (MSCs) could be found in rat cerebral cortex. Methods Rat cerebral cortex samples were diges-ted with typeⅡ collagenase into single cells. Culture medium for MSCs was used for cell culture. The mor-phology of the cells was observed under a microscope. MSC-specific surface markers were detected by flow cytometry. Differentiation potential favoring adipogenesis or osteogenesis of the subcultured cells was ana-lyzed. Lymphocyte transformation test (LTT) was performed to analyze their immunosuppressive function. Bone marrow MSCs (BM-MSCs) were used as positive control cells in all experiments. Results The fibro-blast-like cells isolated from the cerebral cortex of rats were morphologically heomeogeneous after cell culture and grew in a spiral pattern. On the surface of these cells, CD29, CD90 and CD146 were highly expressed at mRNA level, while the expression of CD45 and CD31 at mRNA level were not detected. Moreover, they could be induced to differentiate into adipocytes and osteoblasts. Results of LTT showed that they were func-tionally equivalent to BM-MSCs in inhibiting the proliferation of spleen lymphocytes induced by concanavalin A. Conclusions This study suggested that the cells with the characteristic of MSCs could be detected in rat cerebral cortex.
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Objective To explore the effects of diet and/or exercise intervention on oxidative stress of overweight or obese adolescents.Methods Thirty-one adolescents with normal body weight(normal control group)and 93 overweight or obese adolescents(average age(13.6 ±0.7)years; body mass index (BMI)22.4 to 34.1 kg/m2)were enrolled in this study.The overweight or obese participants were then randomly assigned to the diet group(group A),exercise group(group B),diet plus exercise group(group C),and non-intervention group(group D).Individualized caloric intake was carried out.One-hour afterschool exercise was performed once per day,4 days per week for 10 weeks.Changes of anthropometry,body composition,and metabolic biomarkers were determined.Results Compared to the normal control group,serum levels of fasting glucose,total cholesterol(TC),low-density lipoprotein cholesterol(LDL-C),and malondialdehyde(M DA)of the in overweight group were significant increased(all P ＜ 0.05),although the level of SOD was significant decreased(P ＜ 0.05).Serum MDA level was positively correlated with bodyweight,BMI,TC,and LDL-C(R values were 0.209,0.228,0.274,and 0.263,respectively ; all P ＜0.05),and serum SOD level was negatively correlated with bodyweight and BMI(R values were-0.334 and-0.362,respectively ; both P ＜ 0.05).Group A,B and C showed a significant decrease in BMI,waist circumference and body fat after the intervention(all P ＜ 0.05).The level of MDA was significantly decreased after the intervention in group A and B(both P ＜ 0.05); the levels of TC and LDL-C were significantly decreased in group B and C(both P ＜ 0.05).Conclusion:Oxidative stress could have existed in overweight adolescents even when their serum glucose and lipid profiles are at a normal level.Ten-week diet and/or exercise intervention may contribute to improved body weight control,lipid metabolism,and oxidative stress response in this population.