1.Target regulation of miR-9 to the expression of NRP1 and its role in radiation effects
Haiqin ZHANG ; Juancong DONG ; Hui GAO ; Siyao ZUO ; Linlin JIN ; Libo LIU ; Shunzi JIN
Chinese Journal of Radiological Medicine and Protection 2014;34(10):725-728
Objective To explore the effect of miR-9 on the expression of NRP1 and its radiation effects in A549 cells.Methods Bioinformatics was used to analyze the potential binding sites of has-miR-9 and NRP1-3'UTR.The miR-9 sequence was inserted into pcDNA-DEST-47 plasmid to construct the eukaryotic expression vector (pcDNA-DEST-miR-9) and to construct the NRP1 gene 3'UTR luciferase reporter plasmid (pEZX-MT05) at the same time.They were simultaneously transferred into A549 cells for analysis of the regulatory effect of miR-9 on the expression of NRP1.Meanwhile miR-29b was used as a negative control to observe whether or not NRP1 gene was a target of miR-9.After 10 Gy irradiation,the expression of NRP1,and the inhibitory effect of miR-9 on it was confirmed by Western blot assay.The expression of miR-9 was detected by real-time PCR.Results It was found that miR-9 reduced the luciferase activity of NRP1-3'UTR wild plasmid (t =3.906,P < 0.05) but not NRP1-3' UTR mutant plasmid.This luciferase activity was not inhibited by other types of miRNA (miR-29b).The expression of NRP1 protein in A549 cells was decreased after the cells were transfected with miR-9 mimic.After irradiation with dose of 10 Gy,the expression of miR-9 were decreased (t =37.319,P < 0.05) and the expression of NRP1 protein were increased.Conclusions miR-9 regulates the expression of NRP1 by targeting 3'UTR site of NRP1 gene in A549 cells.
2.Analysis of Th1-Th2-Th3 related gene expressions in the thymus of mice irradiated with different doses
Hui GAO ; Siyao ZUO ; Zhengji HUANG ; Hailing HAN ; Juancong DONG ; Haiqin ZHANG ; Shunzi JIN
Chinese Journal of Radiological Medicine and Protection 2015;35(4):248-251
Objective To analyze the effect of high and low dose radiation on the expressions of Th1,Th2 and Th3 /Tr1 related-genes in mice thymocytes and investigate the possible underlying molecular mechanism.Methods ICR mice were randomly divided into low-dose group (0.075 Gy),high-dose group (2.0 Gy) and sham-control group.The mouse thymus tissue was extracted at 16 hours after irradiation and the expressions of Th1-Th2-Th3 related genes were measured by PCR array.Results Eight genes were up-regulated and five genes were down-regulated after low dose radiation (0.075 Gy);while 54 genes were up-regulated and three genes were down-regulated after high dose (2.0 Gy) radiation.These genes included Th1 cell related genes,Th2 cell related genes,Th3/Tr1 cell related genes,Th1/Th2 immune response genes and transcription factor related genes.Low dose radiation induced up-regulation of Stat4 and Socs1 of genes related to the Th1 cells,and it induced down-regulation of IL-4ra,Cebpb,Gata3 and Tgfb3 associated with Th2 and Th3 cells,which lead to Sftpd genes up-regulation of Th1 immune response eventually.The high dose radiation up-regulated all of Th1,Th2 and Th3/Tr related genes and also enhanced the expressions of Cd86,IL-18,IL-10 and Irf4 genes related to Th2 immune response,but it did not alter the gene expression of Th1 immune response.Conclusions Low-dose radiation induces Th1-type immune response,while high doses radiation triggers Th2 type immune response.
3.Effects of endogenous sulfur dioxide on pulmonary vascular inflammation in rats with monocrotaline-induced pulmonary hypertension
Xiaodan JIANG ; Chen LIANG ; Shuxu DU ; Siyao CHEN ; Junbao DU ; Hongfang JIN
Chinese Journal of Applied Clinical Pediatrics 2015;30(1):55-58
Objective To investigate the effects of endogenous sulfur dioxide(SO2) on pulmonary vascular inflammation in rats with monocrotaline (MCT)-induced pulmonary hypertension.Methods Thirty-two Wistar rats were randomly divided into 4 groups(n =8 for each group):control group,MCT group,MCT + L-aspartic acid-β-hydroxamate(HDX) group,and MCT + SO2 group.Rats in the MCT group,MCT + HDX group,and MCT + SO2 group were subcutaneously injected with MCT(60 mg/kg) on the first day.For rats in MCT + HDX group,HDX(25 mg/kg,on day 0,7 and 14) was given orally after injection of MCT; and rats in MCT + SO2 group were subcutaneously injected with the SO2 donor sodium sulfite/sodium bisulfate(Na2SO3/NaHSO3,and mole ratio was adjusted to approximately 3:1) each day.Rats in the control group received only the same volume of solvent vehicle only.After 3 weeks,mean pulmonary artery pressure(mPAP) of each rat was evaluated by using a right cardiac catheterization procedure.Immunohistochemistry was used to detect the expression of inflammatory related factor intercellular adhesion molecule-1 (ICAM-1) and the key molecules of nuclear factor-κB (NF-κB) signal transduction pathway,including p65 and inhibitor of NF-κB (IκBα) in the small pulmonary artery endothelial cells.Results The differences in mPAP,expression of ICAM-1,IκBα and p65 in the small pulmonary artery endothelial cells were found among the 4 groups (mPAP:F =53.334,P < 0.01 ; ICAM-1:F =183.82,P < 0.01 ; IκBα:F =142.89,P < 0.01 ; p65:F =105.46,P <0.01).The mean pulmonary artery pressure(mPAP) was significantly raised in MCT group rats as compared with that of the control group along with upregulated expressions of ICAM-1 protein and p65 protein in small pulmonary artery endothelial cells,while the expression of IκBα protein in small pulmonary artery endothelial cells was significantly low.After administration of HDX,the mPAP and the expression of ICAM-1 protein and p65 protein in small pulmonary artery endothelial cells further increased compared with those of MCT group,while the expression of IκBα protein in small pulmonary artery endothelial cells was significantly lower than that of MCT group.Whereas with treatment of SO2 derivatives,the mPAP,the expression of ICAM-1 protein and p65 protein in small pulmonary artery endothelial cells were significantly lower than those of MCT group,while the expression of IκBα protein in small pulmonary artery endothelial cells increased significantly compared with that of MCT group.Conclusions Endogenous SO2 might inhibit the activation of NF-κB pathway in the small pulmonary artery endothelial cells,attenuate the pulmonary vascular inflammation and prevent the MCT-induced pulmonary hypertension in rats.
4.Case Reports of Two Patients with Pregnancy-associated Breast Cancer and Related Literature Review
Siyao JIN ; Zhigang ZHAO ; Jiping HUO
China Pharmacy 2020;31(8):989-996
OBJECTIVE:To provide reference for the early diagnosis and selection of treatment regimens of pregnancy- associated breast cancer (PABC). METHODS :The disease characteristics ,treatment process and prognosis of 2 cases of PABC were analyzed in our hospital. The relevant literature published from Jan. 1986 to Apr. 2019 in PubMed database was retrieved. The case reports that the title ,keywords or abstracts involved “Breast cancer and pregnancy ”“Pregnancy-associated breast cancer ” “Breast cancer during pregnancy ”“Breast carcinoma during pregnancy ”“Case reports ”were included. Cases which didn ’t meet the definition of PABC were excluded. The general information ,tumor clinical characteristics ,drug treatment plan ,maternal/fetal prognosis and other information of patients were extracted for summary and descriptive statistical analysis. RESULTS & CONCLUSIONS:Two patients were both diagnosed during lactation. The prognosis was good after neoadjuvant chemotherapy and surgical resection. A total of 36 case reports were obtained through literature search and screening ,as well asclinical data of 45 patients(39 diagnosed during pregnancy and 6 diagnosed during lactation ).Neoadjuvant chemotherapy AC regimen (doxorubicin+ cyclophosphamide)was used in 35.0%(14/40)of cases after excluding the cases without relevant information ;elective caesarean section was performed in 59.5%(22/37)of cases ,37.8%(14/37)of cases were delivered ,and 1 case chose to terminate pregnancy;survival rate of patients was 80.8%(21/26),and the average weight of newborns was 2 407 g(1 015-3 830 g). Six patients each received taxanes during pregnancy and 9 patients during postpartum. The determination of chemotherapy for PABC should comprehensively consider a variety of factors. It is necessary to comprehensively weigh the benefit risks of the mother and child,try to avoid chemotherapy in early pregnancy ,and especially consider the impact of chemotherapy on the fetus. The chemotherapy regimen is still dominated by anthracyclines. Based on this ,an individualized regimen is formulated and close monitoring should be performed when using paclitaxel.