1.Effect of mfn2 on mitochondrial function in steatosis hepatocytes
Yong ZHANG ; Wenjun HU ; Yao WANG ; Yun XIA ; Qichang ZHENG
Chinese Journal of Pathophysiology 2010;26(3):568-572
AIM: To investigate the effect of mfn2 on mitochondrial function in steatosis hepatocytes. METHODS: Plasmid pEGFP-mfn2 was transfected into hepatocyte strain L02 by Lipofectamine 2000 in vitro, then the steatosis model of hepatocytes was establish by oleic acid induction. RT-PCR was used to evaluate mRNA expression and Western blotting was use to detect the protein expression. ATP level was determined by firefly luciferase bioluminescent. ROS production was measured by fluorescence probe DCFH-DA. Chondrosome transmembrane potential of L02 was observed by labeling of JC-1 and FCM. RESULTS: The stable expression of ectogenesis mitofusin2 in L02 cells was confirmed by RT-PCR and Western blotting. In the model of oleic acids induced lipid formation, Mfn2 obviously inhibited the descent of chondrosome transmembrane potential and ATP level, and increased ROS production in L02 cells. CONCLUSION: Up-regulated expression of mfn2 attenuates mitochondria dysfunction caused by oleic acids induced lipid formation.
3.Diagnostic value of MRI for spontaneous intracranial hypotension syndrome
Xiumei XIA ; Peng WANG ; Shuling HE ; Yong ZHENG ; Xiaogen PAN
Chinese Journal of Primary Medicine and Pharmacy 2014;(10):1455-1457
Objective To analyze the magnetic resonance imaging ( MRI ) appearances of spontaneous intracranial hypotension syndrome ( SIHS) ,in order to improve the understanding of this disease .Methods The clini-cal data of 7 patients with SIHS confirmed by lumbar puncture were retrospectively analyzed .MRI was performed in all cases,in which there were 6 cases with contrast-enhanced MRI.Results All of them showed diffusely thickening of dural.6 cases with brain contrast-enhanced MRI demonstrated linear ,non-nodular and diffusely dural enhancement . The dilation of sinus and great vein were observed in 5 cases,subdural hydroma in 2 cases,brain sagging in 3 cases, pituitary gland plumpness in 2 cases,subdural hematoma in 1 case and 1 case with linear spinal dural enhancement . Conclusion The MRI manifestations of SIHS are characteristic ,and it will be helpful for the diagnosis by combining clinical data.
4.Preparation of 131I-K237 and the experimental study on targeting therapy in nude mice bearing human lung cancer
Huo-qiang, WANG ; Jun-yong, XIA ; Zhi-yong, ZHANG ; Long, ZHAO ; Jin-xu, ZHENG
Chinese Journal of Nuclear Medicine 2010;30(6):390-394
Objective To establish the radiolabeling method for peptide K237 with 131I and investigate the biodistribution and therapeutic efficacy of 131I-K237 on nude mice bearing human lung cancer.Methods Iodogen method was used for labeling K237. The bioactivity of 131I-K237 was tested by human umbilical vein endothelial cell ( HUVEC ) proliferation inhibitory assay and the affinity of 131I-K237 was examined by competition binding studies. Twenty-five mice were divided into five groups randomly, including physiologic saline (group 1), K237 (40 μg) (group 2), 131I ( 11. 1 MBq) (group 3), 131I-K237 (K237 40 μg, 11. 1 MBq) intravenously ( group 4), and 131I-K237 ( K237 40 μg, 11.1 MBq) intratumorally (group 5). Injections were repeated at 15 d after the first injection. The tumor growth inhibition rate was calculated. Student's t-test and analysis of variance (ANOVA) were used for testing significant differences of data. Results The inhibition rate of HUVEC proliferation had no significant difference between radiolabeled K237 and unlabeled K237 ( (73.69 ± 5.36) % vs ( 62.68 ± 3.83 ) %, t = 1.67, P > 0.05 ). The growth of transplanted lung cancer was inhibited by 75. 01 % in group 4, 78.99% in group 5, 31.15% in group 2 and 12.61% in group 3, respectively. The average tumor volume of groups 4 and 5 were significantly smaller than that of groups 1,2, and 3 ( F = 15. 233 and 13.611, respectively, P <0. 01 ). Conclusion 131I-K237 can be readily radiolabeled and it can effectively inhibit the growth of tumor in nude mice bearing human lung cancer.
5.Hypothesis of 'saturated or nonsaturated cytotoxicity model' clarifies antibodies mediated immunoreaction in transplant recipient
Bicheng CHEN ; Hui GUO ; Yong LIU ; Sheng CHANG ; Yong CAI ; Peng XIA ; Shaoling ZHENG ; Yirong YANG
Chinese Journal of Organ Transplantation 2011;32(6):351-354
Objective To develop the hypothesis ‘saturated or non-saturated cytotoxicity model' and explain the various phenomena of antibody mediated immunoresponses in recipients,including rejection and accommodation.Methods The imitating complement dependent cytotoxicity.The threshold set to identify as saturated or non-saturated cytotoxicity depends on antigen-antibody complex(R)whether or not above lethal number(D)in effective time.Feasibility of the hypothesis was examined through explaining various phenomena mediated by anti-donor antibodies,especially some contradictory phenomena.Results Hyperacute rejection,accelerated rejection and acute rejection could be well explained by saturated cytotoxicity.Accommodation of ABO imcompatible transplantion,de novo antibody induced injury,change of protein profile,and C4d deposition in graft could be well elucidated by the hypothesis.Conclusion The hypothesis saturated or nonsaturated cytotoxicity model' help to interpret and interconnect various phenomena of antibodies mediated immune response,such as rejection and accommodation.
6.Progress of studies on genetics of childhood acute leukemia.
Journal of Experimental Hematology 2013;21(3):791-795
This study on determination of leukemia-specific chromosomal abnormalities and their relationship with prognosis of childhood acute leukemia (AL) had an important significance for childhood acute leukemia. In recent years, the efficacy of treatment of childhood AL has been greatly improved, but relapse is still a main factor affecting prognosis. Treatment based on the risk stratification by cytogenetic abnormalities can improve the prognosis and survival rate. In the past 3 decades, the genetic techniques have developed rapidly and many new genetic abnormalities have been found. This review highlights the main chromosomal and genomic abnormalities of 3 common childhood AL, including B-cell precursor acute lymphoblastic leukemia (BCP-ALL), T-cell acute lymphoblastic leukemia (T-ALL) and acute myeloid leukemia (AML).
Acute Disease
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Child
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Humans
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Leukemia
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genetics
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Leukemia, Myeloid, Acute
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genetics
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Precursor B-Cell Lymphoblastic Leukemia-Lymphoma
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genetics
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Precursor T-Cell Lymphoblastic Leukemia-Lymphoma
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genetics
7.Hydrodynamics-based transfection of IL-10 and TGF-β1 plasmids prolonging the graft survival in murine skin transplant model
Peng XIA ; Yirong YANG ; Wenwei CHEN ; Shaoling ZHENG ; Yong CAI ; Longren XIAO ; Bicheng CHEN
Chinese Journal of Organ Transplantation 2009;30(3):144-147
Objective To investigate the protective effect of hydrodynamics-based injection with plasmids of IL-10, TGF-β1 and TGF-β1 + IL-10 in murine skin transplantation model. Methods Plasmids were constructed by inserting coding sequences of IL-10 and TGF-β1. In F1 mice (Balb/c×C57BL/6, H-2b/d) to Balb/c (H-2d) murine skin transplant model, 20 μg plasmid (blank or IL-10 or TGF-β1 or IL-10 + TGF-β1) was injected to donors by hydrodynamics-based method in first day and every interval 2 days for 6 times. The survival of grafts was observed after 7 days of transplantation. After C56BL/6 spleen cells transfused Balb/c accepted 5 times hydrodynamics-based injection as above,CD4+ CD25+ T regulatory cells of spleen were measured by FACS. Results The survival time of graft in each group was (13.50±1.04)days (blank group), (13.83±1.16)days (IL-10 group), (15.33±1.50) days (TGF-β1 group), and (21.33±3.20) days (IL-10 + TGF-β1 group),respectively (P<0.05). The percentage of CD4+ CD25+ cells was (6. 58±1.86)% (blank group),(10.52±1.13)% (IL-10 group),(14.44±0.42)% (TGF-β1 group),and (14.25±1.24)% (IL-10+TGF-β1 group) respectively (P<0.05). Conclusion Hydrodynamics-based transfection of IL-10 combined with TGF-β1 can synergistically enhance the percentage of CD4+ CD25+ T cells and prolong the graft survival.
8.The application of universal stem loop primer for microRNA scanning and quantification
Bicheng CHEN ; Silu WANG ; Yongheng BAI ; Yunxiu YANG ; Yong CAI ; Peng XIA ; Shaoling ZHENG ; Yirong YANG
Chinese Journal of Laboratory Medicine 2011;34(10):926-930
Objective To establish a universal stem loop primer (USLP) based real-time PCR method to scan mature miR profile and quantify it's expression.Methods The common universal stem-loop primer pairs were re-designed; 8 random nucleotides were introduced at 3 ' end for reverse transcription of the mature miR,establishing a miR scanning and quantifying system based on SYBR Green Ⅰ PCR (improved USLP method).10-fold gradient diluted standard miRNA-155 cDNA ( 1 ~ 109 copies/μ1) were utilized to evaluate the sensitivity of this method.The specificity was verified by melting curve assay; the precision was assessed by intra-assay coefficient of variation (ICV) of threshold cycle (Ct value) through 20 repeated detections of the standard miR-155 cDNA (2 × 105,2 × 106,2 × 107 copies/μl) ; cost of the primers and time were evaluated,compared with that of the conventional USLP method.Peripheral blood samples were cultured with phytohaemagglutinin (PHA) for0 h,16 h,24 h,48 h and 72 h,and 87 candidate miR that may be associated with human immunity from PubMed data were scanned and quantified from the cultured T cells.Results The sensitivity of the improved USLP method was 103 copies/μl of standard miR-155 cDNA.Melting curve assay showed a single melting peak at 80 ℃,suggesting the excellent PCR specificity of miR-155.Precision of our method quantifying miR-155 was acceptable (ICV < 2.5% ).Compared with the traditional stem loop primers,our method saved 75% cost of primers ( 1 917 bp vs 7 851 bp) and 60% test time of reverse transcription (85 min vs 205 min).By our method,85 of the 87 miR expression in T cells had no significant difference after the PHA stimulation; the expression of miR-150 (72 h) decreased by 10 times and that of miR-155 (48 h) increased 8 times after culture with PHA (Z =-2.032,P =0.042;Z =- 2.023,P =0.043,respectively ).Conclusions The improved USLP method is fast,precise,sensitive,and cost-effective.It could be used for miR profile scanning and quantifying in T cells.
9.Evaluation of misoprostol on cervix in hysteroscopy
Huan-Xia LU ; Chao-Sheng ZHENG ; Mei-Ling XIE ; Yong-Feng ZHANG ;
Chinese Journal of Primary Medicine and Pharmacy 2006;0(07):-
Objective To study the effect of misoprostol used to intenerate cervix and ease pain in hys- teroscopy.Methods 380 cases were divided into two groups randomly;260 cases of them were placed misoprostol 200?g in vagina 2 hours before hysteroscopy and 120 cases treated without drugs served as control.The diameter of cerical,hemorrhage,the rate of PAAS and bellyache were observed.Results The diameter of cerical,hemorrhage. the rare of PAAS and bellyache in the group of misoprostol were lower than those in the group of antitheses(P
10.Hepatoprotective Effects of Silybin-Phosphatidylcholine Complex in Septic Rats
xing-xia, LIU ; pei-yong, QIU ; jian-zhi, GAO ; zheng-yue, CHEN
Journal of Applied Clinical Pediatrics 2004;0(09):-
Objective To investigate the hepatoprotective effect of silybin-phosphatidylcholine complex(SPC) in sepsis rats.Me-(thods) Fifty Wistar immature rats were randomly divided into 3 groups:normal control(10 cases),septic group(30 cases) and interfe-(rence) group(10 cases).In septic group and interference group,rats were treated by cecal ligation and puncture(CLP).Meanwhile,the interference group was given SPC before 2 hours of CLP and after 2 hours of CLP.Blood of rats was taken from all groups to determine the levels of serum tumor necrosis factor-?(TNF-?),interleukin-1(IL-1),alanine aminotransferase(ALT) and aspartate aminotransferase(AST) at varying intervals.Results The levels of serum TNF-?,IL-1,ALT and AST after CLP were significantly elevated in septic group compared with the normal control group(P