ObjectiveTo explore the changes of astrocytes in vitro on the expression of Cyclin D1 under normal and hypoxia/reoxygenation conditions.MethodsThe primary astrocytes were isolated from the cortex of SD fetal rats(less than 24 hours) and identified by immunosytochemical method with anti-GFAP antibody after 3 passages.Astrocytes of passage 3 were divided into normal group ( control group),hypoxia/reoxygenation group ( H/R 37℃ ),and mild hypothermiaintervention group( H/R 32℃ )separately.Astrocytes from the later tow groups were reoxygenated with 4,12,and 20 hours separately after exposed to hypoxia conditions for 8 hours.Trypan blue staining was employed to detect the survival rates and immunofluorescence,western-blot were used to analyse the expressin of Cyclin D1 of of astrocytes of different groups and time points.Results 1.The GFAP positive astrocytes from passage 3 exceeded 95 ％.2.With regard to morphology and survival rates,there is no difference between astrocytes of normal and hypothermia groups after 8 hours exposure to hypoxia conditions.Reoxygenation could obviously rise astrocytes mortality with time went by ( H/R 37 ℃ group:12.87 ± 2.76 ( R4 ),31.55 ± 3.00 ( R12 ),46.40 ±8.50(R20) ;H/R 32℃ group:6.77 ± 1.53( R4),15.97 ±4.00(R12),28.33 ±5.69(R20) ;all P＜0.05).3.Immunofluorescence and western-blot revealed that reoxygenation increased Cyclin D1expression markedly,which was proportional to the duration of reoxygenation.Mild hypothermia could reduce Cyclin D1 expression of astrocytes severely under reoxygenation condition.ConclusionCyclin D1 expression can be regarded as a sensitive index of damage to astrocytes caused by hypoxia/reoxygenation conditions.

Objectives To evaluate the count of endothelial progenitor cells (EPCs) in peripheral blood (PB) and bone marrow (BM) of acute leukemia (AL) patients and explore its clinical significance.Methods EPCs were detected by flow cytometry procedures in 43 AL patients and in 10 benign hematologic patients as control group.Results The absolute counts of EPCs in AL patients before the treatment [(119.46± 72.23)/μl in BM and (13.69±8.26)/pl in PB] were significantly higher than those in control group [(23.21 ± 12.59)/pl in BM and (1.86±1.18)/μl in PB] (P ＜ 0.01).The absolute counts of EPCs were significandy higher in BM than those in BP in AL patients before the treatment (P ＜ 0.001).After the treatment, the absolute counts of EPCs in no remission (NR) group [(110.02±67.28)/μl in BM and (10.04±9.51)/μ1 in PB] were significantly higher than those in control group (P ＜ 0.05), while the counts of EPCs in complete remission (CR) group were no significant difference compared with those in control group (P ＞ 0.05).After the treatment ,the absolute counts of EPCs both in BM and in BP of CR group [(26.32±17.44)/μl and (2.54±2.12)/μl, respectively] were significantly lower than those before treatment [(113.18±69.22)/μl and (14.45±10.76)/μl, respectively] (P ＜ 0.05), however those of NR group were no significant difference than before (P ＞ 0.05).The absolute counts of EPCs whether in PB or in BM were no significant difference between acute myeloid leukemia (AML) and acute lymphoid leukemia (ALL) (P ＞ 0.05).The absolute counts of EPCs in PB of AL had a positive correlation with β2-MG and LDH (P ＜ 0.05).Conclusions EPC levels are significantly increased in BM and BP of AL patients and may correlate with disease status, response to treatment and prognosis.

ObjectiveTo explore psychological and behavioral characteristics of children with autism spectrum disorder (ASD) using Psycho-educational Profile (Third Edition) (PEP3). MethodsFrom October, 2021 to October, 2022, 192 children with ASD without intervention in the Binzhou Medical University Hospital were selected as observation group, and 96 healthy children who visited at the same time were selected as control group. They were assessed with PEP3. ResultsThe development of receptive language significantly delayed behind expressive language (t = 5.383, P < 0.001) in the observation group. The cognitive, expressive language, receptive language, fine motor, gross motor, imitation and personal self-care correlated with age (r = 0.540 to 0.795, P < 0.001). The Cronbach's α of PEP3 for the observation group was 0.810 to 0.947. The original score of each subtest of PEP3 was less in the observation group than in the control group (|t| > 4.267, P < 0.001). ConclusionThe development of receptive language retards behind the expressive language in children with ASD. Cognitive and motor functions develop with age, while the correlation between maladaptive behavior and age is weak. PEP3 is reliable in internal consistency, and valid in discrimination.

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.