1.Regulation of RPL4 promoter activity by transcription factor Runx1
Han ZHANG ; Huifang GAO ; Huyong ZHENG
Chinese Journal of Cancer Biotherapy 2006;0(05):-
Objective:Previous genome-wide microarray analysis found TTCATTCT motif(DNA binding site of Runx1)in the RPL4 promoter of childhood leukemia cells,suggesting that Runx1 may regulate the expression of RPL4.This study is to investigate the relationship between Runx1 and RPL4 so as to better understand the effects of Runx1 on RPL4gene transcription,laying a foundation for further studying leukemogenesis of childhood leukemia.Methods:The luciferase plasmids containing RPL4 promoter and its mutant were constructed and were co-transfected into 293T cells with the expression plasmid of Runx1.The transactivity of RPL4 promoter was assayed by luminometer.Results:Runx1 significantly decreased the transcriptional activity of RPL4 promoter(P0.05).Conclusion:Runx1 can inhibit RPL4 gene transcription in a dose-dependent manner through binding to the TTCATTCT motif.
2.Purification of SEB by McAb affinity chromatography and activity analysis
Jing ZHANG ; Huifang WANG ; Chuan GAO ; Yunyang SONG
Chinese Journal of Immunology 2001;0(10):-
Objective:To prepare highly purified Staphylococcal enterotoxin B(SEB) by affinity chromatography and test its activities.Methods:Anti-SEB McAb(1D2) purified by precipitation method with caprylic acid was coupled to Sepharose 4B. And then the SEB was isolated using an affinity chromatography column. In addition, we analyzed the superantigen activity and antigen activity of SEB.Results:The purification efficiency of SEB was 60.71% by affinity chromatography. Its purity was higher than those of standard preparation and the SEB purified by ion change chromatography. At the same time, the purified SEB by affinity chromatography possesses favourable activities of superantigen and antigen.Conclusion:McAb affinity chromatography could be used for purification of SEB with high efficiency.
3.Mutations in E154 of Diphtheria Toxin (DT) and Their Biologic Activity
Chuan GAO ; Huifang WANG ; Jing ZHANG ; Yunyang SONG ; Yalan HU
Progress in Biochemistry and Biophysics 2006;0(06):-
According to the results of quantum chemistry calculation and the present research status in the relationship between the structures and the functions of DT, the E154 in DT catalyzing domain was mutated to aspartic acid and arginine in order to study the effects of the alteration on the biological activities. By means of gene site-direct mutation, two mutated genes were prepared and the high performance expression was obtained in E.coli system. The results of toxcity studies indicated that the acute toxicity in guinea pig and cytotoxicities of mutant E154D increased slightly in compared with those of recombination wild toxin, and contrarily, those of E154R decreased obviously.
4.The comparison research of intrathecal injection of oxygen and hyperbaric oxygen on the rehabilitation of patients with craniocerebral injury
Ligui GAO ; Huifang WANG ; Wei SHI ; Ruizhi WANG
Journal of Xi'an Jiaotong University(Medical Sciences) 2004;0(05):-
Objective To investigate the role of intrathecal i njection of oxygen in the treatment of patients with craniocerebral injury throu gh the comparison research of intrathecal injection of oxygen and hyperbaric oxy gen on the rehabilitation of patients with craniocerebral injury. Methods A total of 150 patients were divided into three gro ups: the group of intrathecal injection of oxygen, the group treated with hyperb aric oxygen and the group of intrathecal injection of air. CSF was applied via l umbar puncture. The rehabilitative function of patients was observed and the lev el of superoxide dismutase (SOD), malondialedhyde (MDA), and nitrogen monoxide ( NO) in CSF were measured. Results The neurological function of the patients treated w ith oxygen injected intrathecally intrathecal injection of oxygen was improved w as more significanty than that of patients treated with hyperbaric oxygen; the i ncrease of SOD, NO and the decrease of NO were more significant than that in the group treated with hyperbaric oxygen. Conclusion Intrathecal injection of oxygen can improve the rehabilitative function more significantly than the hyperbaric oxygen therapy in the treatment of craniocerebral injury.
5.Evaluate the Significance of Th1 7 Cells in Systemic Lupus Erythematosus with Cardiovascular Disease
Jingxia YAN ; Lixia GAO ; Xia WU ; Huifang GUO ; Hongtao JIN
Journal of Modern Laboratory Medicine 2014;(6):52-54,58
Objective To explore the role of Th17 cells in systemic lupus erythematosus (SLE)with cardiovascular disease (CVD).Methods 61 patients of SLE were collected from September 2011 to March 2013 in the Second Hospital of Hebei Medical University by revised SLE classification standards of ACR in 1997.These patients were divided two groups:22patients of SLE with CVD and 39 patients of SLE without CVD;the control group include 20 healthy.Th17 cells were measured by flow cytometry,IL-1 7 was detected by enzyme-linked immunosorbent assay.The correlation among them and the disease active index were analyzed.Results ①The percent of Th1 7 cells in the group of SLE with CVD,that in the group of SLE without CVD and that in control group were (2.09±0.98)%,(1.75±0.75)% and (0.89±0.44)%,respec-tively.The percent of Th1 7 cells in healthy group were lower than that in SLE with CVD and SLE without CVD group (t=4.717~5.030,P<0.001).The level of IL-17 in the group of SLE with CVD,that in the group of SLE without CVD and that in control group were 85.64±20.76 pg/ml,75.25±28.14 pg/ml and 35.06±6.58 pg/ml respectively,and the serum of IL-17 in healthy group were lower than in SLE with CVD and SLE without CVD group (t=6.275~9.954,P<0.001). There were no significant difference of Th1 7 cells and IL-1 7 between SLE with CVD and SLE without CVD groups (t=1.520,P>0.05;t=1.513,P>0.05).②The level of IL-17 were correlated positively with SLEDAI and the anti-double strand DNA (r=0.393,P=0.008;r=0.558,P<0.001),were correlated negatively with complement C3 (r=-0.423,P=0.005).The percent of Th17 cells in CD4+T cells were correlated positively with SLEDAIand the anti-double strand DNA (r=0.681,P<0.001;r=0.492,P=0.015)were correlated negatively with complement C3 (r=-0.534,P=0.027).Con-clusion The level of Th1 7 cells and IL-1 7 were high in SLE,and they were related with the disease activity.The cardiovas-cular factor had not affect the expression of Th1 7 cells and IL-1 7 in SLE.
6.EXPERIMENTAL OBSERVATION OF TRANSPLANTATION OF CULTURED NEWBORN PIG PANCREATIC ISLET CELLS
Hanmei GOU ; Huifang WANG ; Wei JIANG ; Li XU ; Hui GAO ;
Journal of Xi'an Jiaotong University(Medical Sciences) 1982;0(01):-
In this article, we report the pancreatic islet cells function of newborn pig pancreatic tissues cultured by Ham F_(10), and the condition of graft survived into the bodies of Sprague-Dawley rats, and also study the influence of Glycyrrhiza uralensis to the grafts. The cultured islet cells of newborn pig were transplanted into the mesenteric cavities (the first & second group), kidney cystic(the third group) and brain tissucs (the fourth group) of the 4 groups adult Sprague-Dawley rats. The first group rats drank Glycyrrhiza uralensis water, after 8~11weeks of operation a examination pathological was made with the cut graft The results showed that the first and second group grafts survived over 11weeks, and survived better the group drunk the Glycyrrhiza uralensis water was much better than other. The purpose of this experiment is to find the heteropolar source of pancreatic tissues offering body.
8.Dyslipidemia and important organ damages in patients with active systemic lupus erythematosus
Huifang GUO ; Chenxing PENG ; Mingfeng ZHANG ; Fuling SHAO ; Lixia GAO ; Qing ZHANG ; Fengzhen LIU
Chinese Journal of Rheumatology 2010;14(1):33-36
Objective To investigate the correlation between dyslipidemia and important organ damage in patients with active systemic lupus erythematosus (SLE) without treatment. Methods Serum sam-ples from 71 active SLE patients and 30 healthy controls were obtained to measure lipid profiles including total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL), low-density lipoprotein cholesterol (LDL), apolipoproteinA1 (apoA1), apolipoproteinB100 (apoB) and lipoprotein a (LPa). Clinical parameters were recorded. Results The levels of serum TC, TG, LDL, apoB in active SLE patients were higher than those in healthy controls, in contrast,the levels of HDL,apoA1 were much lower (P<0.05 or P<0.01). Patients with important organ damages had longer disease course and elevated levels of serum TC, TG, LDL and apoB concentrations than those without organ damage (P<0.05 or P<0.01), especially in patients with cadiovascular diseases (CVD) (P<0.01). Moreover, these changes in lipid metabolism were positively correl-ated with disease course and negatively with C3 level (P<0.05 or P<0.01). The elevated serum TC and LDL concentrations were negatively correlated with C4 level (P<0.05). Conclusion Severe dyslipidemia is present in active SLE patients.It is correlated with disease course and disease activity. Increased serum TC, TG, LDL and apoB concentrations play key roles in patients with important organ damages.
9.Abnormal expression of T regulatory cells in primary Sj(o)gren's syndrome
Lixia GAO ; Fengchun ZHANG ; Yumei MA ; Xiaolei LIU ; Huifang GUO ; Hongtao JIN
Chinese Journal of Rheumatology 2012;16(7):477-480,封3
Objective This study was designed to investigate the expression of T regulatory cells (Treg) in the peripheral blood and salivary gland of patients with primary Sj(o)gren's syndrome (pSS).Methods The expression of Treg was measured by flow-cytometry analysis in 23 pSS patients and 15 healthy controls.CD4 positive cells were sorted by immunomagnetic beads,Forkhead box protein P3 (Foxp3) mRNA was extracted,real time polymerase chain reaction (RT-PCR) was used to measure Foxp3 mRNA.For the study of salivary gland,five primary billary cirrhosis (PBC) patients without pSS were selected as the control group,the expression of CD4+T cells and Foxp3+ T cells in salivary gland was examined by immunohistochemistry.The result was analyzed by t test.Results The expression of Treg in the peripheral blood of pSS patients [(5±6)%] was lower than healthy controls [(10±5)%] (t=2.190,P=0.036).There was no significant difference in Foxp3-mRNA expression between pSS patients (0.08±0.05) and controls(0.09±0.03 ) (t=0.695,P>0.05 ).The expression of CD4+ T cells [ ( 30± 10 )% ] and Foxp3+ T cells [ ( 10.7±5.8 ) % ] in the salivary gland of pSS patients increased significantly when compared with PBC control group [CD4+T (11±6)% and Foxp3+T(3.2+1.1)% ] (t=4.072,2.840; P<0.05).Conclusion For pSS patients,the expression of Treg decreases in the peripheral blood,but Treg increases in the salivary gland.The results of this study suggest that Foxp3plays an important role in the pathogenesis of pSS.
10.Pathogenic role of Bcl-6/Blimp-1/IL-21 in primary Sj?gren′s syndrome
Chunqiao DUAN ; Wenwen YANG ; Lingyan LEI ; Huifang GUO ; Mingfeng ZHANG ; Lixia GAO ; Chao SUN
Chinese Journal of Immunology 2015;(3):384-388,392
Objective:To analyse the role of Bcl-6/Blimp-1/IL-21 in pathogenesis of primary Sj?gren′s syndrome(PSS),the relationship between Bcl-6/Blimp-1/IL-21 expression and labial gland biopsy grading.Methods:Immunohistochemical(IHC) method was used to detect the expression of Bcl-6/Blimp-1 in salivary gland between 30 cases pations with PSS in disease group and 11 cases patients with mucous cyst,lower lip trauma in control group,and the serum IL-21 levels between disease group of 40 cases patients with PSS and 30 cases healthy volunteers were measured by enzyme-linked immunosorbent assay( ELISA) ,relations among 15 patients with IL-21 expression levels with labial gland biopsy grading.Results:The expression levels of Bcl-6/Blimp-1/IL-21 in disease group were higher than control group;in disease group,with pathological grades increased,the expression levels of Bcl-6/Blimp-1/IL-21 were also raised.Conclusion:①Bcl-6/Blimp-1/IL-21 may participate in the pathogenesis of PSS;Bcl-6/Blimp-1/IL-21 is associated with infiltration lymphocytes of salivary gland.