4.The differential gene expression in rat lymphocytes after exposure to 2 Gy γ-rays
Hui YUAN ; Jianguo LI ; Jingjing YIN ; Xiujun QIN ; Gang LI
Chinese Journal of Radiological Medicine and Protection 2016;36(12):898-903
Objective To investigate the impacts of γ-ray radiation on gene expressions of peripheral blood lymphocytes in SD rats,and to screen differential expression genes and biological pathways closely related to radiation injury.Methods The differential gene expression of peripheral blood lymphocytes in SD rats was selected by microarray at 6 h after 2 Gy 60Co γ-ray exposure in vitro and in vivo.Bioinformatics analysis of the differentially expressed genes was performed by using Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) databases.Real-time quantitative PCR was applied to verify the screen results of the microarray.Results Fifty-five genes with three times over-expressed level were screened out from the radiation groups both in vitro and in vivo and they were involved in 6 signaling pathways.There were two differentially expressed genes of microarray assay were verified by PCR assay.Conclusions Differential expressed genes in the peripheral blood lymphocytes of SD rats could be induced by γ-ray radiation and they cooperatively contributed to a variety of biological processes.
5.Relationships between induction of apoptosis by CDDP in Scaber cell and apoptosis-related proteins
Kui WU ; Gang MENG ; Yuan WANG ; Qin ZHOU
Chinese Pharmacological Bulletin 2003;0(10):-
AIM To investigate the effect of cisplatin on apoptosis in Scaber cell. METHODS The apoptotic cells were detected by TUNEL,HE,eletronic micrpscopy. RESULTS Treatment of Scaber cells with CDDP resulted in characteristics typical of apoptosis. CDDP induced apoptosis of Scaber cells in time and concentration dependent manner. To further investigate the mechanism of apoptosis induced by CDDP, the expressions and activity of apoptosis associated proteins such as bcl 2, bax and caspase 3 were examined using S P method.The results showed: CDDP caused time and concentration dependent decreases in bcl 2 and increased in bax proteins.CDDP bcl 2 and its translocation to perinuclei and nuclei. The expression of caspase 3 in Scaber cell were determined during apoptosis induced by CDDP. CONCLUSION Our investigetion showed that the apoptosis induced by CDDP is related to the increase of bax protein, and the decrease of bcl 2 protein. and its translocation to perinuclei and nuclei.
6.MRI demonstrates abnormalities of motor nerves and extraocular muscles in congenital fibrosis syndrome due to mutations in KIF21 A
Yong-Hong, JIAO ; Shao-Qin, WU ; Feng-Yuan, MAN ; Hong-Yan, JIA ; Gang, LIU ; Nan, LIN
International Eye Science 2014;(8):1426-1429
AIM:To observe the structural basis of ocular motility abnormalities in patients with congenital fibrosis of the extraocular muscles type Ⅰ ( CFEOM Ⅰ) due to missense mutations in the developmental kinesin KIF21A using high - resolution magnetic resonance imaging ( MRI) .
METHODS: Totally 11 affected individuals reported KIF21A mutations were correlated with MRI studies demonstrating extraocular muscles ( EOMs ) size, location, contractility, and innervation. EOMs and the motor nerve in the orbits were imaged with T1 weighted in a triplanar scan using a dual-phased coils with 2. 0mm thick. Motor nerves were imaged at the brainstem using head coils and 3D-FIESTA with 0. 6-mm thick.
RESULTS: Patients with CFEOM Ⅰ exhibited different degrees of hypoplasia of oculomotor nerve, the abducens nerve and the trochlear nerve were also affected, of which 8 cases of orbital section could see the signal of abnormal nerve dominated by oculomotor nerve to lateral rectus. The both sides of six EOMS in all patients exhibited variable atrophy and abnormal bright internal signal on T1 imaging, particularly severe for the superior rectus and levator muscles.
CONCLUSION: High - resolution MRI can directly demonstrate pathology of motor nerves,affected EOMs, and ‘Pulley' hypoplasia caused by CFEOM Ⅰ due to mutations in KIF21A,and these findings suggest that the neuronal hypoplasia is the etiological factor of CFEOM.
7.The effects of nitric oxide synthase and its antagonist on alkali burn-induced corneal neovascularization
Gao-qin, LIU ; Yuan, CHEN ; Lei, CHEN ; Yan-hui, XIAO ; Zhi-gang, CHEN ; Pei-rong, LU
Chinese Journal of Experimental Ophthalmology 2013;31(10):908-913
Background Though nitric oxide (NO) and NO synthase (NOS) have a critical role in angiogenesis,their effects on corneal neovascularization (CNV) and mechanism need to be further explored.Objective The aim of this study was to explore the effects of NOS and its antagonist,Nw-nitro-L-arginine methyl ester (L-NAME) on experimental CNV in mice,and investigate the influence of NOS and L-NAME on the tube formation of human retinal endothelial cells (RECs) in vitro.Methods The CNV models were established in the left eyes of 36 male BALB/c mice aged 7-8 weeks by application of the filter paper with NaOH in the center of corneas.The mice were randomized into two groups.L-NAME of 10 g/L (0.5 ml) was intraperitoneally injected 1 week before induction of CNV three times a week for three weeks in the mice of the L-NAME injection,and PBS was used in the same way in the control group.CNV was examined under the slit lamp biomicroscope 2,4,7,14 days after NaOH burn.The expression of CD31 in the CNV was assayed to calculate the ratio of CNV area and total corneal area using whole mount technique.The expression of NOS mRNA in the corneal tissue was detected by reverse transcriptase polymerase chain reaction (PCR),and VEGF expression in the human RECs was assayed by Western blot.The vessel formation number of cultured human RECs with or without L-NAME was performed by matrigel in vitro.Grouped t test was used to compare the differences of the parameters between the two groups.Results CNV developed and peaked 2 weeks after the application of NaOH on the mice corneas,and the CNV was obviously less in the L-NAME group compared with the control group.The expression of NOS mRNA in the corneas (NOS mRNA/ GAPDH mRNA)was significantly lower in the L-NAME group than that of the control group 2,4,7 days after CNV induction (t =19.481,t=22.059,t=10.961,all at P<0.01).The ratio of the CD31 positive area in whole corneal area was 0.59± 0.01 in the L-NAME group,and that of the control group was 0.78±0.10,showing a significant difference between the two groups (t =3.078,P<0.05).Western blot assay showed that the relative expression of VEGF protein in human RECs was declined in the L-NAME group compared with the control group 0,2,4,7 days,with statistically significant differences in 4 days and 7 days after NaOH burn(t=7.696,t=17.953,both at P<0.01).The number of vessel network was 46.33±1.86 in the L-NAME group and 64.00±4.51 in the control group,with a significant difference between them (t =3.623,P<0.05).Conclusions NOS participated in the pathogenesis and aggravation of CNV induced by NaOH.L-NAME arrests CNV formation and human RECs tube formation through down regulating the VEGF expression and NOS activity.
8.Investigation on T-lymphocyte subsets absolute counting and CD4+/CD8+ratio in healthy adults of Ugyur and Han nationalities in Karamy
Qin FENG ; Gang DU ; Guiying YANG ; Lijuan ZHANG ; Xinrong YUAN ; Shanjun ZHU
International Journal of Laboratory Medicine 2015;(22):3227-3228,3231
Objective To built the reference range of peripheral blood T-lymphocyte subsets including CD3+ ,CD4+ ,CD8+ and CD4+ /CD8+ ratio in healthy adults of Ugyur and Han nationalities and to provide basis for the diagnosis ,therapy and prognosis of disease .Methods A total of 181 blood samples were collected from healthy adults .The cell chip quantitative detection technology was used to detect CD3+ ,CD4+ and CD8+ ,CD3+ ,CD4+ absolute value ,CD8+ and CD4+ /CD8+ ratio were compared between Ugyur and Han nationalities .Results CD8+ absolute counting and CD4+ /CD8+ ratio had no significant difference between Ugyur and Han nationalities(P>0 .05) ,while the CD3+ ,CD4+ absolute counting had significant difference(P<0 .05) .Conclusion The discrepancy of territory and living environment should be taken into account for building a reference values of CD4+ .
9.Effect of Triptolide on Expression of Oxidative Carbonyl Protein in Renal Cortex of Rats with Diabetic Nephropathy
DONG XING-GANG ; AN ZENG-MEI ; GUO YUAN ; ZHOU JIA-LIANG ; QIN TAO
Journal of Huazhong University of Science and Technology (Medical Sciences) 2017;37(1):25-29
The traditional Chinese medicine (Tripterygium wilfordiiHook.f.,TWH) has been clinically used to treat primary and secondary renal diseases and proteinuria for nearly 40 years.However,there is a rare literature about the effect of triptolide (the main active ingredient of TWH) on the expression of oxidative carbonyl protein (OCP) in diabetic nephropathy (DN).This study aimed to provide experimental evidence for triptolide treatment on DN through its effect on the expression of OCP,in order to investigate the effects of triptolide on the expression of OCP in rats with DN.Sixty SD rats were randomly divided into five groups:control group,high-dose triptolide (Th) group,low-dose triptolide (T1) group,DN model group,and positive control (benazepril) group.The DN model was established using streptozotocin.Urinary protein excretion,fasting blood glucose (FBG),superoxide dismutase (SOD) in renal homogenate,malondialdehyde (MDA) in renal homogenate and renal nitrotyrosine by immunohistochemistry,and the expression of OCP by oxyblotimmune blotting were detected.In the DN model group,rat urinary protein excretion and renal MDA were significantly increased,while renal SOD significantly decreased and nitrotyrosine expression was obviously upregulated in the kidney.After triptolide treatment,24-h urinary protein excretion (61.96±19.00 vs.18.32±4.78 mg/day,P<0.001),renal MDA (8.09±0.79 vs.5.45±0.68 nmol/L,P<0.001),and nitrotyrosine expression were decreased.Furthermore,renal OCP significantly decreased,while renal SOD (82.50±19.10 vs.124.00±20.52 U/L,P<0.001) was elevated.This study revealed that triptolide can down-regulate the expression of OCP in the renal cortex of DN rats.
10.Relationships between induction of apoptosis by Taxol in Hela cells and apoptosis-related proteins
Xiangyang HU ; Gang MENG ; Yangyi BAO ; Xiaomei ZHU ; Yuan WANG ; Qin ZHOU
Chinese Pharmacological Bulletin 2003;0(09):-
Aim To investigate the effects of taxol on apoptosis in Hela cell and the mechanism of apoptosis. The apoptotic cells were detected by TUNEL, HE, eletronic micrpscopy and flow cytometry method. The expressions and activity of apoptosis associated proteins such as PCNA and caspase-3 were examined using S-P and enzyme histochemistric method.The results followed as: HeLa cells exposed to Taxol undergo cell death, presenting morphological and biochemical characteristics typical of apoptosis and the apoptotic cells increased with time and concentration. In contrast to untreated Hela cells, which express low PCNA, Ones treated with Taxol expressed high amounts of PCNA. Conclusion Taxol may induced apoptosis in Hela cell. The apoptosis induced by taxol is related to the increase of PCNA protein and activity of caspase-3.