ABSTRACT
ObjectiveTo explore the typical behavior characteristics of children with autism in symbolic play and the value of Symbolic Play Test (SPT) in the early identification of autism. MethodsFrom November, 2021 to September, 2022, a total of 260 children with language problems were collected from Department of Children's Health Care of Binzhou Medical University Hospital. A total of 193 children with autism were as observation group and 67 normal children were as control group. All children played symbolic games. The typical behavioral characteristics of children with autism in SPT were explored, and a reliability and validity analysis based on the results of SPT was conducted. They were assessed with the adaptive and personal social scores of Gesell Development Scale, and the correlation of the scores of Gesell Development Scale and the score of SPT was analyzed. ResultsThe Cronbach's α coefficient of SPT of children with autism was 0.835 to 0.935, and the total score of SPT, the scores of surrogate object, fictional attribute and fictional object were positively correlated with each other (r > 0.607, P < 0.001). The SPT scores decreased in the observation group (t > 9.615, P < 0.001), and SPT score positively correlated with adaptability and personal-social development quotient (r > 0.609, P < 0.001). ConclusionTypical behavior of children with autism can be reflected in symbolic play, and SPT can provide clues for early identification of autism.
ABSTRACT
Objective To breed homozygous mice of Treg-Th17 fluorescence double labeling and to investigate the role of Treg-Th17 balance in endotoxemia. Methods Mice of Treg and Th17 fluorescence single labeling were brought from Harvard Medical School,USA and were mated and bred based on the genetic rules.The genomic DNA was extracted from the tails of second generation weaning mice for genotyping by polymerase chain reaction (PCR).Thereafter,the homozygous mice of fluorescence double labeling were selected to have successive inbreeding for three generations and their genotypes and growth status were measured.The endotoxemia model in vivo was duplicated and changes of Treg-Th17 balance were detected by flow cytometry analysis at 24 h and 48 h after endotoxemia. Results Among the seven second generation mice in the first batch,only three (two males and one female) were the wanted homozygotes.Meanwhile,the genotypes of their offsprings for the next three generations were all Foxp3 RFPKI-homo IL-17A GFPKI with normal growing status.As important component of endotoxin,lipopolysaccharide (LPS)could induce significant signal expressions of red fluorescence protein (RFP) and green fluorescence protein (GFP).The ratio of CD8a + Foxp3 +,CD8a + IL-17A +,CD4 + Foxp3 + and CD4 + IL-17A + in the lymphocytes went up significantly in the group of 24 hours of LPS treatment,as compared with the control group (P ＜ 0.01 or P ＜ 0.05 ),while the difference between the control group and the group of 48hours of LPS treatment had no statistical significance (P ＞ 0.05). Conclusion Foxp3 RFPKI-homoIL-17A GFPKI mice have identical genotypes and stable genetic characteristics and is a model animal and novel research platform that can provide real-time monitoring of the changes of Treg-Th17 balance in vivo.The role of T reg-Th17 balance works significantly at 48 hous after the subject is exposed to LPS stimulation,and the roles of CD8a + Foxp3 and CD8a + IL-17A + in Treg-Th17 balance require further investigation.
ABSTRACT
Objective To investigate the action mechanism of glutamate-mediated signaling pathway in malignant melanoma. Methods WM451LU melanoma cells in log phase were classified into 6 groups, negative control group treated with PBS (100 μl), MK801 group treated with the non-competitive N-methyl-D-aspartate (NMDA) receptor antagonist MK-801 (100 μmol/L), CPCCOEt group treated with non-competitive metabotropic glutamate receptor 1 (mGluR1) antagonist CPCCOEt, MAP2 group transfected with adenovirus vector containing microtubule associated protein 2a (Ad-MAP2a), MK801 + MAP2 group treated with MK801 of 100 μmol/L and transfected with Ad-MAP2a, CPCCOEt + MAP2 group treated with CPCCOEt of 10 μmol/L and transfected with Ad-MAP2a. Western blot was performed to detect the expression of an ionotropic glutamate receptor, i.e., N-methyl-D-aspartate receptor type 2A (NMDAR2A) in WM451LU cells transfected with Ad-MAP2a. Scratch motility assay and cell invasion assay were conducted in vitro to detect the changes in migration and invasion ability of WM451LU cells after treated with Ad-MAP2a, MK-801, CPCCOEt alone or in combination. In vivo study was carried out to compare the inhibitory effect of the above treatments on melanoma. Results Western blot revealed a decrease in the expression of NMDAR2A in WM451LU cells after transfected with Ad-MAP2a. The scratch motility assay showed that the number of migrating cells per high power field was 117.04 ± 2.76 in MAP2 group,107.64 ± 6.50 in MK801 group,97.36 ± 4.79 in CPCCOEt group, 43.28 ± 3.02 in MK801 + MAP2 group,30.76 ± 3.97 in CPCCOEt + MAP2 group,significantly different from that in the negative control group (152.3 ± 5.75,all P ＜ 0.01 ). Cell invasion assay demonstrated that the average number of invading cells per high power field in the negative control was significantly higher than that in MAP2 group, MK801 group, CPCCOEt group, MK801+MAP2 group and CPCCOEt + MAP2 group (170.43 ±8.72 vs. 98.26 ± 3.84, 97.22 ± 5.54, 112.23 ± 7.21, 42.89 ± 5.06, 58.25 ± 6.68, P ＜ 0.05, 0.05, 0.05, 0.01and 0.01, respectively).A significant decrease was observed in the average volume of experimental melanoma in mice of MAP2 group, MK801 group, MK801 + MAP2 group, CPCCOEt group and CPCCOEt + MAP2 group compared with the negative control group (224.02 ± 46.19 mm3, 160.33 ± 33.91 mm3, 91.49 ± 21.48 mm3,202.30 ± 52.37 mm3, 111.13 ± 69.81 mm3 vs. 342.70 ± 60.92 mm3, all P ＜ 0.01 ). Conclusions To block the glutamate signaling pathway in vitro can inhibit the invasion and migration of melanoma cells, and to block the pathway in vivo can inhibit the growth of malignant melanoma and alter the morphology of melanoma cells.