1.Construction and erpression of NDV pIRHN nucleic acid vaccine and its effect onthe tumor cells
Lijuan XUE ; Wei GONG ; Tao GE
Chinese Journal of Immunology 1985;0(05):-
Abstract Objective:To explore antitumor effect of NDV HN. Methods: pIRHN nucleic acid vaccin was constructed and was transfectedto Hela cells. Westem blot was used to analyssize the expression of pIRHN nucleic acid vaccine in eukarytic cell. The mode of cell death was de-tected by fluorescence microscope, gel electrophoresis and TUNEL assay. The effect of pIRHN nucleic acid vaccine on the contents of siaha acidin the Hela cell was examined. Results: pIRHN nucleic acid vaccine could be expressed in the eukarytic cell. pIRHN could induced apoptosisafter HeLa cells were transfected. The effect of antitumor responses of pIRHN was correlated with the contents of sialic acid in tumor cells. Con-clusion: The reults of our experiment will provide the theory evidence for the anti tumor mechamism of NDV.
2.A Study on Apoptosis and Caspase-3 Expression in the Immune Response of Inner Ear
Lijuan XU ; Shusheng GONG ; Jibao WANG
Journal of Chinese Physician 2001;0(02):-
Objective To investigate whether apoptosis is invovled in the immune response of inner ear,and relation between the signal transduction of Caspase-3 and apoptosis. Methods Sixteen healthy,female guinea pigs were employed in this study. The animals in the experiment group were sensitized systematically with keyhole limpet hemocyanin(KLH),then were inoculated in inner ear with the same antigen, and the animals in the control group were injected with the same volume of PBS. The animals were sacrificed at the 5th day after inner ear vaccination. Paraffin sections of cochleas from animals were prepared, TUNEL assay was used to detect apoptotic cells in inner ears, and immunohistochemical method was applied to detect the expression of Caspase-3 in inner ears. Results TUNEL-positive cells are found in the inner ears of the experiment group, but did not in the inner ears of the control group except for a few TUNEL-positive cells in the supporting cells, the stria vascular cells and the spiral ganglion cells. The high expression of Caspase-3 could be detected in the inner ears of the experiment group, but no positive cells of Caspase-3 were found in the inner ears of the control group. Conclusion Apoptosis could be induced by the immune response of inner ear, and Caspase-3 cascade may be involved in the course of apoptosis.
3.Development of a rapid triplex TaqMan real-time PCR assay for the detection of toxigenic Vibrio cholerae
Hui YANG ; Cheng GONG ; Lijuan WU ; Jihong HU ; Jianping CAI
Chinese Journal of Laboratory Medicine 2009;32(9):1038-1043
for the detection of their virulent abilities simultaneously.
4.Prestin Expression in HEI-OC1 Cells With Oxidative Stress Damage
Junyi WANG ; Xiaomin LAI ; Lijuan GONG ; Yuan XIA ; Zhi WANG
Journal of Audiology and Speech Pathology 2017;25(1):49-52
Objective To examine the effects of oxidative stress induced damage to the Prestin expression in HEI-OC1 cells,and to study the mechanism of sensory deafness.Methods We used different concentrations (50μM,100μM,200μM)of hydrogen peroxide canister to cultivate HEI-OC1 cells,and to detect the activity of su-peroxide dismutase(SOD).The quantitative real-time PCR and immunofluorescence were used to detect the prestin expression of mRNA.Results The SOD activity decreased in the HEI-OC1 cells damaged by oxidative stress.The high concentration of the infected group decreased more significantly(F= 9926.293,P<0.01).The expressions of Prestin mRNA and Prestin protein were decreased obviously in the HEI-OC1 cells.The high concentration of in-fected group decreased more significantly (F= 4065.046and7657.217,P<0.01).Conclusion Oxidative stress in-ducing damage inhibits the expression of prestin.Prestin protein may be used as a molecular marker of sensory deafness.
5.The mitochondrial Na+/Ca2+exchanger may reduce high glucose-induced oxidative stress and nucleotide-binding oligomerization domain receptor 3 inflammasome activation in endothelial cells
Yuan ZU ; Lijuan WAN ; Shaoyuan CUI ; Yanping GONG ; Chunlin LI
Journal of Geriatric Cardiology 2015;(3):270-278
Background The mitochondrial Na+/Ca2+exchanger, NCLX, plays an important role in the balance between Ca2+influx and efflux across the mitochondrial inner membrane in endothelial cells. Mitochondrial metabolism is likely to be affected by the activity of NCLX because Ca2+activates several enzymes of the Krebs cycle. It is currently believed that mitochondria are not only centers of energy produc-tion but are also important sites of reactive oxygen species (ROS) generation and nucleotide-binding oligomerization domain receptor 3 (NLRP3) inflammasome activation. Methods&Results This study focused on NCLX function, in rat aortic endothelial cells (RAECs), induced by glucose. First, we detected an increase in NCLX expression in the endothelia of rats with diabetes mellitus, which was induced by an injection of streptozotocin. Next, colocalization of NCLX expression and mitochondria was detected using confocal analysis. Suppression of NCLX expression, using an siRNA construct (siNCLX), enhanced mitochondrial Ca2+influx and blocked efflux induced by glucose. Un-expectedly, silencing of NCLX expression induced increased ROS generation and NLRP3 inflammasome activation. Conclusions These findings suggest that NCLX affects glucose-dependent mitochondrial Ca2+signaling, thereby regulating ROS generation and NLRP3 in-flammasome activation in high glucose conditions. In the early stages of high glucose stimulation, NCLX expression increases to compensate in order to self-protect mitochondrial maintenance, stability, and function in endothelial cells.
6.Clinical and epigenetic study of a case with adrenal hypoplasia congenita caused by a novel DAX-1 gene mutation
Yuping GONG ; Guang XING ; Baoan WANG ; Lijuan YANG ; Jingtao DOU ; Yiming MU ; Juming LU ; Changyu PAN
Chinese Journal of Endocrinology and Metabolism 2009;25(1):62-63
Hormones and epigenetic characteristics in a patient with clinically diagnosed adrenal hypoplasia congenita (AHC) were analyzed. Results indicated that plasma ACTH increased, while cortisol, testosterone, LH and FSH decreased. LH, FSH and testosterone did not sufficiently respond to GnRH or hCG stimulation. Gene analysis indicated that C368F mutation was located in exon 1 of DAX-1 gene.
7.Fast violet B salt staining for bone marrow stromal cells and its clinical significance
Xubo GONG ; Xianguo WU ; Xingguo LU ; Lijuan YAN ; Xibin XIAO ; Genbo XU ; Xiaohong ZHANG ; Xiaoying ZHAO
Chinese Journal of Laboratory Medicine 2008;31(8):890-893
Objective To establish a quick method to identify BMSC by fast violet B salt staining and evaluate the clinic value. Methods Smears of separated and cultured BMSC, bone marrow, pleural and ascitic fluids were made, then the staining of fast violet B salt was performed. The BMSC in aplastic anemia (AA), high hyperplasia and normal groups were counted and compared with each other. Meanwhile, the diagnostic value of this method to AA was evaluated. Results The cytoplasm of BMSC presented mauve, while the nucleus were negative, other cells such as myelocytes, nucleated erythrocytes, megakaryocytes, monocytes, macrophages, lymphocytes and plasmacytes were negative. The count of BMSC in AA, high hyperplasia and normal group was 1.07 ± 0. 29, 2. 26 ± 0. 37 and 1.58±0. 33, respectively. Significant differences were found between AA and high hyperplasia groups, AA and normal groups, high hyperplasia and normal groups, respectively (P < 0.01). The sensitivity, specificity, positive likelihood ratio and negative likelihood ratio of this method for diagnosis of AA were 90%, 93%, 12. 86 and 0. 11,respectively. Conclusions The fast violet B salt staining is simple and convenient. It could be used to identify BMSC and play an important role in judging the hyperplasia extent and differentiation of AA.
8.A quantitation method for andrographolide and dehydroandrographolide by X-ray powder diffraction Fourier fingerprint pattern technique
Ningbo GONG ; Lijuan Lü ; Chao LIU ; Lin MA ; Ruoyun CHEN ; Yang Lü
Acta Pharmaceutica Sinica 2010;45(5):673-6
The powder X-ray diffraction Fourier fingerprint pattern technique was used to develop a new quantitation method for the analysis of andrographolide and dehydroandrographolide. And the high performance liquid chromatography method was used to evaluate the quantity of andrographolide and dehydroandrographolide. The relationship of diffraction peak intensity and content of andrographolide and dehydroandrographolide was investigated. The powder X-ray diffraction Fourier fingerprint pattern analysis technique can be used to evaluate the quantity of andrographolide and dehydroandrographolide in the herb simultaneously.
9.Effect of energy-enriched formula for catch-up growth in malnourished infants after congenital heart disease surgery
Yanqin CUI ; Lijuan LI ; Na ZHOU ; Jiangbo QU ; Chunmei HU ; Jianbin LI ; Sitang GONG
Chinese Journal of Clinical Nutrition 2017;25(3):176-182
Objective To investigate the efficacy and safety of enriched-calorie formula in post-operative infants with congenital heart disease and malnutrition.Methods All malnourished infants less than 6 months diagnosed congenital heart disease: ventricular septal defect and had undergone surgery in Guangzhou Women and Children`s Medical Center from December 1,2014 to May 30,2015 were included in this study.All cases were randomly divided into intervention group(energy-enriched formula,intervention group)and control group(standard formula,control group)for enteral nutrition intervention and observed for 3 months.Body mass,body length,upper arm circumference,blood prealbumin(PA),retinol binding protein(RBP),and B-terminal pro-brain natriuretic peptide(NT-proBNP)were measured before and after ICU,after discharge,and 1 month and 3 months after operation.Results Fifty-one cases were in intervention group and 50 cases in control group,respectively.There were no significant differences in body mass,body length,arm circumference,PA,RBP,mean enteral nutrition starting time,mechanical ventilation time,length of ICU stay,hospitalization time,and average fluid intake between the two groups(all P>0.05).The average caloric intake in intervention group was significantly higher than in control group [(437.24±6.68)kJ vs.(312.43±86.22)kJ,P=0.001].There was no significant difference in NT-proBNP,PA,and RBP at different time points between the two groups(all P>0.05).The improvement of nutrition in intervention group was significantly higher than that in control group at 1 month(25.0%vs.4.9%,P=0.011)and 3 months(64.1%vs.15.7%,P<0.001)after operation.Body mass increased in intervention group [(0.067±0.348)kg] compared with that in control group,and decreased [(0.125±0.425)kg] in control group(P=0.015).Body weight[(5.46±1.36)kg vs.(4.80±1.01)kg,P=0.008],weight for age Z score(WAZ)(-2.79±1.28 vs.-3.75±1.27,P<0.001),and height for weight Z score(WHZ)(-2.47±1.43 vs.-3.62±1.77,P=0.001)one month after surgery were significantly higher than those before operation.Body weight [(6.78±1.42)kg vs.(5.72±1.01)kg,P<0.001] arm circumference [(12.80±1.17)cm vs.(12.00±0.90)cm,P<0.001],WAZ(-1.60±1.17 vs.-3.10±1.40,P<0.001),height for age Z score(HAZ)(-1.41±1.63 vs.-2.10±1.41,P=0.034),and WHZ(-0.86±1.31 vs.-2.59±2.13,P<0.001)of the two groups at 3 months postoperative were significantly higher than those before operation,and the growth rate of intervention group was faster than control group.There were no significant adverse reactions in both groups.Conclusion Calorie-enriched formula powder can help malnourished infants to catch up after congenital heart disease surgery.
10.Comparison Researches on the Mode of Cell Death Induced by NDV-Strain Changchun and NDV-Strain Siping
Wei GONG ; Ningyi JIN ; Lijuan XUE ; Qinfang LUO ; Dahui SUN ; Tao GE ; Ping LI ;
Chinese Journal of Cancer Biotherapy 1995;0(02):-
Objective: To investigate the mode of cell death caused by NDV strain Changchun and NDV strain Siping. Methods: Plaque formation, cell suppression test, gel electrophoresis, and TUNEL assay were used after the cells were infected by NDV. Results: The apparently pathological changes were observed in chicken embryo fibroblasts, BHK, Hela, Hep 2, HCT and OS 732 tumor cells, but not in Wish cells. The higher suppressed effect on tumor cells was found in the NDV strain Changchun than that in the NDV strain Siping. There was no dose effect relationship between NDV and tumor cell suppression, only optimum dose NDV could cause maximal tumor cells inhibitory effect. Conclusion: The mode of cell death might be different after infection of NDV. The NDV strain Changchun killed tumor cells mainly through apoptosis, while the NDV strain Siping killed tumor cells mainly through necrosis. \[