1.Glioma stem cell clones and molecular genetics characteristics of primary and recurrent gliomas
Jia SHI ; Xuchen DONG ; Xiaoxiao DAI ; Haiyang WANG ; Xingliang DAI ; Jiachi LIU ; Qianqian JIANG ; Yujing SHENG ; Jun DONG
Chinese Journal of Neuromedicine 2019;18(9):865-874
Objective To explore the molecular genetic characteristics of primary and recurrent glioblastomas (GBMs) from the same patient in vivo, primary glioma stem cells cultured in vitro, and patient-derived xenograft (PDX). Methods (1) The primary and recurrent GBM specimens from one patient during surgical resection were collected; and the expressions of glial fibrillary acidic protein (GFAP), nestin and Ki-67 were detected by immunohistochemical staining; the methylation of O6-methylguanine DNA methyltransferase (MGMT) gene, mutation of isocitrate dehydrogenase (IDH) gene and amplification of epidermal growth factor receptor (EGFR) gene were analyzed. (2) The primary and recurrent GBM stem cells were cultured in vitro and named as SU5-1 and SU5-2 cells, respectively; the expressions of nestin and CD133 were detected by immunohistochemical staining; GFAP expression was detected by immunohistochemical staining after induced differentiation, and the growth curve was detected by CCK-8 assay; Transwell invasion assay was used to detect the invasion ability; cell resistance to temozolomide (TMZ), carboplatin (CBP), cisplatin (DDP) and adriamycin (ADM) was detected by CCK-8 assay; the protein expression of programmed death receptor-ligand 1 (PD-L1) was detected by Western blotting. The rate of PD-L1 positive cells was detected by flow cytometry; genetic testing analysis was as above. (3) The primary and recurrent in situ PDX models in nude mice were established, and the expressions of nestin, GFAP and Ki-67 were detected by immunohistochemical staining. Results (1) As compared with the primary GBM, the recurrent GBM had significantly higher percentages of Ki-67 and nestin positive cells, while statistically lower percentage of GFAP positive cells (P<0.05); genetic analysis showed that there was no mutation in IDH gene in the primary GBM tissues and recurrent GBM tissues; the MGMT gene in the primary GBM tissues was methylated and EGFR gene was not amplified, while the MGMT gene in recurrent GBM tissues was demethylated and EGFR gene amplification was positive. (2) Both SU5-1 and SU5-2 cells expressed nestin and CD133, and GFAP was expressed after induced differentiation; the growth curve showed that the proliferation of SU5-2 cells started earlier than that of SU5-1 cells, the two were equal on the 3rd, 4th, and 5th d, and the proliferation of SU5-1 cells was faster than that of SU5-2 cells from the 6th d; the invasion ability of SU5-2 cells was statistically stronger than that of SU5-1 cells (P<0.05); the inhibition rates of SU5-2 cells treated with 5, 10, and 15 mmol/L CBP, 0.3125, 1.25, and 5 mmol/L DDP, 0.5 and 2 mmol/L ADM, and 125 and 500 mmol/L TMZ were significantly lower than those of SU5-1 cells treated with the same concentrations and same drugs (P<0.05); the protein expression of PD-L1 in SU5-2 cells was higher than that in SU5-1 cells; the positive rate of PD-L1 in SU5-2 cells was statistically higher than that in SU5-1 cells (P<0.05); the results of genetic analysis were consistent with those of the primary and recurrent GBM samples. (3) As compared with those in the primary PDX model, the nestin and Ki-67 expressions were significantly higher and GFAP expression was significantly lower in the recurrent PDX model (P<0.05); the results of genetic analysis were consistent with those of the primary and recurrent GBM samples. Conclusions Genetic differences are detected between primary and recurrent GBMs; recurrent GBM has stronger invasive capacity and multi-drug resistance. The primary stem cells derived from surgical specimens and corresponding PDX models could replicate the molecular genetic characteristics of original tumors, which provide a reliable experimental platform for both tumor translation researches and screening of molecular therapeutic targets.
2.The mechanism of lipoxin A4 intervention on the oxidative stress induced by uric acid in human umbilical vein endothelial cells
You ZHOU ; Jingguo ZHOU ; Tinging YI ; Xiaolan GUO ; Xingliang JIANG
Chinese Journal of Rheumatology 2017;21(4):252-257,后插1
Objective To investigate the effect and mechanism of lipoxin A4 (LXA4) on uric acid (UA) induced oxidative stress of human umbilical vein endothelial cells (HUVECs).Methods The HUVECs was treated with uric acid to constructing the model of oxidative stress,and intervene the model with LXA4 and xylene based iodine (DPI),rotenone.Reactive oxygen species (ROS) of HUVEC were detected by a fluorescence probe 2',7'-dichlorofluorescin diacetate (DCFH-DA).The activity of reduced nicotinamide adenine dinucleotide phosphate (NADPH) oxidase and p47phox protein was measured by Lucigenin enhanced chemiluminescence and Western blotting among control,uric acid (UA),LXA4 and UA +LXA4 groups,respectively.All the results were described by the relative expression of the control group,repeated measure variance analysis and least significant difference test (LSD) were used for statistical analysis.Results UA could stimulate HUVEC to generate ROS with different concentrations and times (F=7.286,F=4.532,P<0.05).Compared with the control group(100±l 1),the ROS production of group with 80 mg/L UA (177±18),120 mg/L (226±29) and 160 mg/L (225±16) increased significantly (t=4.127,t=7.591,t=7.236,P<0.05).Compare with baseline(100±8),the ROS production increased significantly (t=3.688,t=3.513,t=4.526,t=8.269,t=3.829,P< 0.05) at 3 h(143±16),6 h(140±17),12 h(183±20),24 h(240±29) and 48 h(160±22).LXA4 could inhibit ROS generation at different concentrations and times (F=4.008,F =4.497,P <0.05).Compared with LXA4 concentration of 0 nmol/L,the LXA4 concentrations of 10 nmol/L (162±16) and 100 nmol/L (132±15) could significantly inhibit ROS generation(t=3.712,t=4.083,P<0.05).Compared with pretreatment (269±39),the ROS generation decreased significantly (t=6.373,t=6.426,t=7.125,t=6.981,P<0.05).with LXA4 pretreated for 15 min (160±16),30 rain(158±21),1 h (136±13) and 2 h(140±13).Compared with the UA group(252±31),LXA4 and DPI could significantly inhibited ROS generation (145±29,154±27;t=6.356,t=5.853,P<0.05),but Rot was not significantly intervented (241±32;t=1.027,P>0.05).The NADPH oxidase activity in the UA group was significantly higher than that in the control group (144±16,100±13;t=3.659,P<0.05),but the group of LXA4+ UA was significantly lower than that of the UA group (119±14;t=3.124,P<0.05).The cytoplasmic expression of NADPH oxidase subunit p47phox of UA group was significantly lower than that in the control group (47±6,100±8;t=7.562,P<0.05),but the LXA4+UA group was significantly increased compare with the UA group (83±6,t=5.386,P<0.05).The cytomembrane expression of p47phox of UA group was significantly higher than that in the control group (328±36,100±4,t=12.817,P<0.05),but the LXA4+UA group was significantly decreased compared with the UA group (183±30,t=5.129,P<0.05).Conclusion LXA4 inhibits UA induced ROS production in HUVECs.This mechanism might be through inhibiting p47phox trafficing from cytoplasm to cytomembrane,results in inhibiting the activation of NADPH oxidase.
3.Effects of lipoxin A4 on the inflammatory reaction induced by uric acid in human umbilical vein endothelial cells
Tingting YI ; Jingguo ZHOU ; Guangcheng LUO ; You ZHOU ; Yufeng QING ; Xingliang JIANG
Chinese Journal of Endocrinology and Metabolism 2015;(9):800-805
Objective To investigate the effects of lipoxin A4 ( LXA4 ) on inflammatory related factors induced by uric acid( UA) in human umbilical vein endothelial cells( HUVECs) . Methods HUVECs were treated with 5, 25, and 50 ng/ml LXA4 prior to exposure to 12 mg/dl UA. Tumor necrosis factor-alpha(TNF-α), interleukin ( IL)-1β, and IL-6 were analyzed with ELISA and realtime PCR. The phosphorylation levels of p38 mitogen-activated protein kinases( MAPK) and NF-κB/p65 were observed with Western blot. Results Stimulation of HUVECs with 12 mg/dl UA markedly increased TNF-α, IL-1β, and IL-6 production(P<0. 01). Pretreatment with LXA4 significantly inhibited UA-induced production of TNF-α, IL-1β, and IL-6 in a concentration dependent manner(P<0. 01). The mRNA expressions of TNF-α, IL-1β, and IL-6 in response to UA were also decreased by LXA4(P<0. 05). Western blot analysis showed that the phosphorylation levels of p38 MAPK and NF-κB/p65 were significantly raised by 12 mg/dl UA in HUVECs(P<0. 05), but attenuated significantly in the presence of 50 ng/ml LXA4. Conclusion LXA4 may inhibit the expressions of inflammatory related factors induced by UA via reducing p38 MAPK and NF-κB/p65 phosphorylation and play a role in anti-inflammation.
4.Prevalence of diabetes and prediabetes in northeast Sichuan area
Guangcheng LUO ; Tingting YI ; Zhen CHAI ; Sulan LIU ; Jiankang DENG ; Xingliang JIANG
International Journal of Laboratory Medicine 2015;(4):480-481,484
Objective To investigate the prevalence of diabetes and prediabetes in northeast Sichuan area.Methods A total of 8 053 adult residents(over 18 years old)from the hospital who had underwent physical examination were recruited in the study.The concentrations of Fasting plasma glucose(FPG),glycated hemoglobin A1c(HbA1C)and blood lipids were measured.Diabetes and prediabetes were defined according to the 2013 American Diabetes Association criteria.Results The FPG and HbA1C concentra-tions increased with age.The overall prevalence of diabetes and prediabetes were estimated to be 8.5% and 39.6% in the adult pop-ulation in northeast sichuan area respectively.The prevalence of diabetes and prediabetes in men were significantly higher than those in women(10.7% vs.5.7%,41.5% vs.37.2%,P <0.05,respectively).Furthermore,the prevalence of diabetes increased with age,and the prevalence of prediabetes had a trend of first enhancement followed by a decline.Conclusion The prevalence of diabe-tes and prediabetes are relatively high in northeast Sichuan area.
5.Reference interval for hemoglobin A1c in healthy adults in northeast Sichuan area
Yan LIANG ; Zhong TANG ; Xingliang JIANG ; Sulan LIU
International Journal of Laboratory Medicine 2014;(19):2650-2651
Objective To establish reference interval of hemoglobin A1c(HbA1c) determined in healthy adults in northeast Si-chuan .Methods Venous blood was pumped from 494 individuals without previously diagnosed diabetes and other critical illness . The HbA1c level ,blood routine examination ,blood lipids ,fasting plasma glucose ,liver and kidney function were measured . Reference value of HbA1c was determined by 95% confidence interval through the mean of HbA1c .Results The HbA1c level took on normal distribution in 494 healthy individuals ,and the reference interval was 4 .482% -6 .012% .There was no statistical signifi-cance of HbA1c level between different genders(t= -0 .905 ,P= 0 .366) .The levels of HbA1c in 20 - < 35 years old people , 35- <65 years old people and ≥65 years old people were(5 .109 ± 0 .150)% ,(5 .224 ± 0 .122)% ,(5 .444 ± 0 .125)% ,and the differences were statistically significant among different age group(P<0 .05) .The HbA1c level and age were positively correlated (r=0 .338 ,P<0 .01) .Conclusion It is necessary to establish appropriate reference intervals of HbA1c for different laboratories or areas .
6.Study on a new urine analysis core module based on semi-reflection mirror.
Longcong CHEN ; Gaiqin LIU ; Nan HU ; Ruiying ZHANG ; Qifeng JIANG ; Bin GAO ; Xingliang XIONG
Journal of Biomedical Engineering 2014;31(6):1288-1293
A new urine analysis core module based on high performance 32-bit microprocessor and high precision color sensor was presented. A novel optical structure and a specific circuit were applied to improve measurement precision and temperature was used to compensate for results in this core module. The information of urine test peice, such as all original data and color RGB value, reflectivity, semi-quantitative level, etc. can be output. The results showed that the measuring precision was about 95% or above with ideal stability and reliability using this presented core module, which can be conveniently applied in various urine analyzers, and can greatly decrease the cost of urine analyzers in development and production.
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Equipment Design
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Microcomputers
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Reproducibility of Results
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Urinalysis
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instrumentation
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methods
7.Application of superposition theorem in teaching transistor amplifier circuit
Longcong CHEN ; Qifeng JIANG ; Bin GAO ; Xingliang XIONG
Chinese Journal of Medical Education Research 2013;(4):401-403
Transistor amplifier is the key and difficult points in the teaching of medical electronics,however,most of the teaching materials on this content is insufficient.Meanwhile,medical students have little knowledge of engineering,so it is difficult for them to grasp the related knowledge.We introduced ‘ superposition theorem’ into the relevant teaching.Teaching practice proved that this reform made it easier for medical students to have an in-depth understanding of the content and better teaching effect was achieved.
8.Association of the rs10489070 polymorphism of SLC2A9 gene with primary gout in Northeast Sichuan of Chinese Han population
Min LI ; Jingguo ZHOU ; Yufeng QING ; Wenguang XIE ; Dan JIANG ; Qibin YANG ; Mingcai ZHAO ; Xingliang JIANG ; Mei ZENG
Chinese Journal of Rheumatology 2012;16(4):233-238
Objective To detect the distribution of SLC2A9 rs10489070 polymorphism genotypes in Chinese Han population,and to explore the association of this gene polymorphism with gout susceptibility,tophi,serum uric acid levels and other clinical and laboratory data.Methods A total of 151 primary gout patients and 176.healthy controls were enrolled into this study.The genotypes and alleles frequencies were calculated by using TaqMan(R) SNP Genotyping Assays and the possible association between gene polymorphism of SLC2A9 and gout was investigated.T test,Chi-square and Fisher exact probabilities were used for statistcal analysis.Results Genotypes distribution were in Hardy-Weinberg equilibrium in gout patients and controls (P>0.05).The frequency of CC genotype in gout patients was significantly higher than that in the controls (78.8% vs 68.5%,P<0.05),aand the frequency of CG genotype in gout patients was significantly lower (19.9% vs 30.1%,P<0.05).However,there were no statistical differences in the alleles frequencies of C and G between gout patients and controls (P>0.05).Interestingly,there was significant difference in the distribution of genotypes between tophaceous gout patients and non-tophaceous gout patients (P<0.05),and the frequency of CG genolype was much lower in tophaceous gout patients (0 vs 22.7%,P<0.05).Conclusion Results of present study suggest that rs10489070 polymorphism of the SLC2A9 gene might be associated with gout development.CC genotype predisposes to gout,and CG genotype might protect Chinese Han population from gout and tophi development.
9.The relationship between the level of serum soluble CD40L and diabetic retinopathy
Xingliang JIANG ; Jingguo ZHOU ; Zhong TANG
Chinese Journal of Diabetes 2008;16(10):614-616
Objective To investigate the relationship of serum soluble CD40L with diabetic retinopathy(DR) in patients with type 2 diabetes mellitus(DM). Methods Serum soluble CD40L level was measured by ELISA in 30 diabetic patients without diabetic retinopathy (NDR), 29 patients with background diabetic retinopathy (NPDR), 31 patients with proliferative diabetic retinopathy (PDR), and 32 normal control(NC) subjects. Results The serum soluble CD40L levels were higher in NDR,NPDR and PDR groups [(4.55±3.66), (6.65±4.24), (8.31±5.23) μg/L,P<0.01] than in NC group[(2.01±1.35)μg/L]. Conclusions Sorluble CD40L level in serum is significantly increased in type 2 DM with diabetic retinopathy,which may be correlated with pathogenesis of DR
10.Double-reagent direct spectrophotometric method for determination of total iron-binding capacity in serum
International Journal of Laboratory Medicine 2006;0(01):-
0.05. The reference range of seum TIBC( ?1.96s) was 51.3-76.3?mol/L(70 cases).Conclusion The method was stable,simple, sensitive, rapid and accurate, and suitable for the routine manul and automatic analysis of total iron-binding capacity in serum.

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