1.Hypoxia preconditioning may protect focal cerebral ischemia in rats by downregulation of glycogen synthase kinase 3β, phosphorylated signal transducer and activator of transcription 3
Zhiwei TANG ; Zongyu XIAO ; Shizheng WU
International Journal of Cerebrovascular Diseases 2017;25(1):61-67
Objective To investigate the roles of phosphorylated glycogen synthase kinase 3β (pGSK3β) and phosphorylated signal transducer and activator of transcription 3 (pSTAT3) in hypoxic preconditioning-induced neuroprotection against ischemic brain injury in rats.Methods Sixty SD rats were randomly divided into a sham operation group,a cerebral ischemia group,and a hypoxia preconditioning group (n =20 in each group).A model of middle cerebral artery occlusion (MCAO) was induced by the modified suture method.Before the preparation of MCAO model,the rats in the hypoxia preconditioning group were put into a hypobaric oxygen chamber at a simulated altitude of 5 000 m (pressure:0.53 × 105 kPa;partial pressure of oxygen:81 mmHg;1 mmHg =0.133 kPa),3 h a day for 5 days.At 24 h after MCAO modeling,the rats were subjected to neurobehavioral score (n =6) and cerebral infarction volume measurement (n =6).Immunohistochemical staining was used to detect the expression levels of neuronal nuclei (NeuN) and pGSK3β (Ser9) (n=7).Western blot was used to detect the expression levels of pGSK3 β (Ser9) and pSTAT3 (Tyr705) in the ischemic cortex (n =7).Results The neurological deficit score (1.833 ±0.408 vs.2.667 ± 0.516;t =3.101,P=0.011) and cerebral infarction volume (18.137% ± 0.801% vs.24.125% ± 0.694%;t =13.840,P< 0.001) in the hypoxia preconditioning group were significantly lower or smaller than those in the cerebral ischemia group.Immunohistochemical staining showed that the numbers of NeuN positive cells in the cerebral ischemia group and the hypoxia preconditioning group were significantly less than that in the sham operation group (48.000 ± 1.414/high power field [HPF],124.833 ± 3.061/HPF,and 213.500 ± 2.429/HPF;F =7 150.550,P < 0.001),the hypoxia preconditioning group was significantly more than the ischemia group (P <0.001);the numbers of pSTAT3 positive cells in the cerebral ischemia group and the hypoxia preconditioning group were significantly higher than that in the sham operation group (57.667 ± 1.366/HPF,29.167 ± 1.941/HPF and 3.500 ± 1.049/HPF;F =1 962.649,P <0.001),and the hypoxia preconditioning group was significantly less than the ischemia group (P <0.001).Western blot analysis showed that the expression levels of ischemic cortical pGSK3β and pSTAT3 in the cerebral ischemia group and the hypoxia preconditioning group were significantly higher than those of the sham operation group (pGSK3 β:2.336 ± 0.102,0.876 ± 0.196 and 0.440 ± 0.012;F =1 610.826,P < 0.001;pSTAT3:8.368± 0.230,4.883± 0.123 and 0.595± 0.138;F=4018.051,P<0.001),the hypoxia preconditioning group were significantly lower than the ischemia group (all P <0.001).Conclusions Hypoxia preconditioning has neuroprotective effect for ischemic brain injury in rats.It may be associated with the down-regulation of the expressions of pGSK3 and pSTAT3.
2.An experimental model of chronic renal allograft rejection in SD-Wistar rats
Pengcheng YU ; Yongguang LIU ; Ying GUO ; Min LI ; Zongyu XIAO ; Konghe HU ; Jinjun HUANG ; Jun XIN ; Zhiqiang WU ; Ming ZHAO
Chinese Journal of Tissue Engineering Research 2015;(40):6520-6525
BACKGROUND:Fisher-Lewis rat kidney transplant models are the international common chronic renal alograft rejection models, but their application is greatly limited because of difficulty in model preparation and high costs. OBJECTIVE:To explore a new method of establishing SD-Wistar rat models of chronic renal alograft rejection. METHODS: Fifty-six pairs of SD-Wistar rats were subjected to left kidney orthotopic transplantation. The right kidneys of the recipients were intact and used as internal controls. 23 rat recipients were randomly divided into model group (n=15) and control group (n=8). The rats in the model group were injected with cyclosporine microemulsion for 10 days (2 mg/kg/day,i.p.) after kidney transplantation. The rats in the control group were not treated with immunosuppressive therapy. RESULTS AND CONCLUSION:The irreversible acute rejection occurred in al the transplanted kidneys of rats in the control group within 4 weeks, leading to the necrosis of transplanted kidney. Moderate inflammatory cel infiltration appeared in the transplanted kidneys of rats in the model group at 4, 8 and 12 weeks after transplantation. Typical histopathological changes of chronic rejection were observed within 12 weeks after transplantation. The Banff total scores were increased with time after transplantation. Al these histopathological changes were not observed in the intact right kidneys of rat recipients in both groups. The valey value of 
3.Status of anemic among kindergarten children within Suzhou urban area from 2018 to 2022
ZHOU Bi, HOU Yuanyuan, YANG Xiao, SHI Zongyu, LI Ying, ZHU Feng
Chinese Journal of School Health 2023;44(9):1324-1327
Objective:
To understand the current situation and trends of anemia among kindergarten children in urban area of Suzhou from 2018 to 2022, so as to provide a theoretical basis for prevention and intervention in anemia among kindergarten children.
Methods:
From March 2023, a total of 24 178 person times of children from 59 kindergartens selected by random number table method were enrolled, and their physical examination data from 2018 to 2022 were collected, including hemoglobin (Hb), height, gender, weight. The period of 2018-2019 was defined as before the COVID-19 epidemic, and period of 2020-2022 was defined as the COVID-19 epidemic. Data were analyzed using the Mann Whitney U test, χ 2 test and Spearman s correlation analysis.
Results:
From 2018 to 2019, the M ( P 25 , P 75 ) of Hb levels of children in nursery, middle, and senior class were 118 (112, 129), 120 (112, 132) and 122 (113, 134)g/L, respectively, which were higher than that of during 2020-2022 [116(110, 123), 117(111, 124) , 119(112, 126)g/L, Z =-10.7, -12.7, -12.9, P <0.05]. A total of 4 584 person times of children were anemic, with a detection rate of 19.0%. The overall anemia detection rate of kindergarten children during 2018-2019 was lower than that in 2020-2022 (15.3% vs 20.7%, χ 2=100.8, P <0.05). The anemia detection rate of kindergarten children in 2022 (24.5%) was higher than that in 2020 (20.6%) and in 2021 (17.0%) ( χ 2=93.9, P <0.05). The anemia prevalence of children in the nursery, middle, and senior class were 13.9%, 14.7% and 17.1% during 2018-2019, 19.3%, 15.9% and 26.6% during 2020-2022, and 17.6%, 15.5% and 23.6% during 2018-2022, respectively ( χ 2=10.7, 204.6, 186.8, P <0.01). There was no statistically significant correlation between Hb values and body mass index (BMI) in boys and girls with anemia, and all children in kindergarten ( r=0.03, 0.03, 0.09, P >0.05).
Conclusion
The prevalence of anemia among kindergarten children in the urban area of Suzhou is relatively high. The COVID-19 pandemic may have increased the risk of anemia among children.
4.Experimental study on regulation of immune effect of brain glioma cells by NKG2D mediated by miR-10b
Gang YUAN ; Hu JU ; Zongyu XIAO ; Wenhui LI ; Lixin CAO ; Chaojie HUI
Chinese Journal of Immunology 2024;40(3):507-512
Objective:To observe the regulatory effect of microRNA-10b(miR-10b)on the immune effect of glioma cells and explore its mechanism.Methods:Human glioma cell U251 was cultured to obtain cells in logarithmic growth stage.The cell suspen-sion was prepared according to the concentration of 1.0×105 cells/ml,and the control group,overexpression group,low expression group and blank group were set up,with 6 wells in each group.The negative control,miR-10b mimics and miR-10b inhibitor were transfected by liposome transfection in control group,overexpression group and low expression group,respectively.The blank group was given the same amount of sterile normal saline.Natural killer(NK)cells from peripheral blood of a healthy volunteer was isolated and cultured.The killing activity of NK cells was detected by MTT method.The expression of NK cell activated receptor(NKG2D)on the surface of NK cells in each group were detected by flow cytometry,and the expression of major histocompatibility complex class Ⅰ chain-related gene A(MICA),UL16 binding protein 2(ULBP2)and UL16 binding protein 3(ULBP3)on the surface of U251 hu-man glioma cells in each group were detected.Results:The transfection efficiency of control group,overexpression group and low ex-pression group were(93.55±2.05)%,(95.67±3.14)%,(94.18±3.26)%,respectively.Compared with control group and blank group,the expression of miR-10b increased in overexpression group and decreased in low expression group,and the difference were statisti-cally significant(P<0.05).There was no significant difference in the expression of miR-10b between control group and blank group(P>0.05).Compared with control group and blank group,the killing activity of NK cells with different effect target ratios in overex-pression group decreased,the expression of NKG2D decreased,the killing activity of NK cells with different effect target ratios in low expression group increased,and the expression of NKG2D increased,and the difference were statistically significant(P<0.05).The killing activity of NK cells in each group increased with the increase of effect target ratio,and the difference were statistically signifi-cant(P<0.05),and there was no significant difference in NK cell killing activity and NKG2D expression between control group and blank group(P>0.05).Compared with control group and blank group,the expression of MICA,ULBP2 and ULBP3 on the surface of human glioma cell U251 in overexpression group decreased,and the expression of MICA,ULBP2 and ULBP3 on the surface of human glioma cell U251 in low expression group increased,the difference were statistically significant(P<0.05),and there was no signifi-cant difference in the expression of MICA,ULBP2 and ULBP3 on the surface of U251 glioma cells between control group and blank group(P>0.05).Conclusion:Inhibiting the expression of miR-10b can increase the expression of NKG2D on the surface of NK cells and MICA,ULBP2 and ULBP3 on the surface of human glioma cell U251,and enhance the killing activity of NK cells against human glioma cell U251.