1.Cloning and Construction of Nucleic Acid Vaccine of FABP Gene cDNA from Echinococcus granulosus
Huifang HAO ; Zhigang WANG ; Zhiwei LI
Chinese Journal of Parasitology and Parasitic Diseases 1987;0(04):-
Specific primers were designed according to published nucleotide sequence of FABP (fatty acid binding protein) gene in the GenBank database. The kozak sequence (CCACC) was introduced at the upstream of initiator. The total RNA was extracted from protoscoleces of Echinococcus granulosus (Inner Mongol isolate). The FABP gene cDNA fragment was amplified by RT-PCR and cloned into pMD19-T vector for sequencing and analyzing. The cloned FABP gene cDNA was with 402bp. The ORF encoded 133 amino acids. The amplified cDNA fragment was subcloned into pCDNA3.1(+)vector. The results showed that the nucleic acid vaccine candidate pcDNA-FABP-NM has been constructed.
2.MR-guided Neurolytic Celiac Plexus Block for Treatment of Upper Abdominal Cancer Pain
Hao SHI ; Huifang CAO ; Hongyu DING
Journal of Practical Radiology 1992;0(11):-
Objective To evaluate the value of MR-guided neurolytic celiac plexus block(NCPB)for treatment of upper abdominal cancer pain.Methods 13 neurolytic celiac plexus blocks were carried out in 12 patients with severe upper abdominal pain caused by malignant tumors.The pain-relieving effect of the block was both evaluated using visual analogue scale(VAS) and analyzed statistically in all cases.Results The placement of the needle MR-guided was easy and accurate,the successful rate of the puncture was 92%.There were no severe complications.The pain before and after the procedure had obvious difference and the pain relief could last for a long time.Conclusion MR-guided NCPB is a simple and effective technique for treatment of upper abdominal cancer pain.
3.Effects of Perinatal Diphenylchloroarsine Chloride Exposure on Livability and Reproductive Function of F1 Generation Rats
Xiaojing LU ; Huifang WANG ; Lanqun HAO
Journal of Environment and Health 1992;0(04):-
Objective To study the effects of perinatal exposure of diphenylchloroarsine chloride(DA)on the livability and reproductive function of F1 generation rats.Methods The pregnant rats were treated with DA by gavage at 0(solvent control),0.63(low dose),0.94(medium dose),1.89(high dose)mg/(kg?day)from day 6 of pregnancy to day 15 of lactation.The general condition,the change of body weight and abnormity of main organs of the F0 pregnant rats and F1 young rats were observed,and the livability and abnormity of reproductive function of F1 young rats were examined.Results The results indicated that compared with the negative control,the increasing amounts of the body weights of the F0 maternal rats and the livability of F1 young rats aged 4 day significantly decreased at 0.94 mg/kg and 1.89 mg/kg,the livability in lactation period obviously reduced at 1.89 mg/kg,and the rate of absorbed fetus in F1 pregnant rats increased at 1.89 and 0.94 mg/kg.Conclusion DA exposure may have an adverse effect on livability and reproductive function of the F1 offspring when the dosage is no fewer than 0.94 mg/kg.
4.Determination of Paraquat in Serum by Ultraviolet Spectrophotometry
Yanyan ZHAO ; Huifang LIU ; Lina HAO
Journal of Environment and Health 1992;0(05):-
Objective To establish a method for determining paraquat in the serum by ultraviolet spectrophotometry and present a quantitative index for paraquat poisoning salvage. Methods The sample was deproteinizated by 20% trichloracetic acid(TAC). 50 ?l microcell was used, the detection wavelength was 257 nm. Results The linearity was within 0.05~50 ?g/ml, r=0.999 9. The average recovery rates were 90.0%-102.4% and RSD=3.9%(n=4), the lowest detection limit was 0.01 ?g/ml. Conclusion The method is simple, rapid, precise and suitable for the determination of paraquat in the serum.
5.Effects of Different Compatibility Ratios of Astragali Radix and Angelicae Sinensis Radix on Vascular Intimal Hyperplasia in Rats
Xiwei PENG ; Huifang YAN ; Juan HUANG ; Jiahuang ZHU ; Hao XU ; Xiaoping HUANG ; Changqing DENG
Chinese Journal of Information on Traditional Chinese Medicine 2017;24(6):56-59
Objective To observe the effects of different compatibility ratios of Astragali Radix and Angelicae Sinensis Radix on vascular intimal hyperplasia; To ascertain the effective compatibility of Astragali Radix and Angelicae Sinensis Radix for antagonizing vascular intimal hyperplasia. Methods SD rats were divided into different groups by baseline geometric design method: Astragali Radix and Angelicae Sinensis Radix different compatibility ratios groups, Astragali Radix group, Angelicae Sinensis Radixgroup, positive medicinegroup and sham-operation group. A model of intimal hyperplasia of thoracoabdominal aorta was established by balloon catheter injury in vascular endothelium of rats. Then the thoracoabdominal aorta was taken out after gavage of Astragali Radix and Angelicae Sinensis Radix with different compatibility ratios for 14 days. Bloodletting was used to take thoracoabdominal aorta. Masson staining and Morphometric methods were used to analyze the intimal hyperplasia. Results IA, IT, HRIA and HRIT increased 14 day after intimal injury. Compared with the model group, IA, IT, HRIA and HRIT in Angelicae Sinensis Radix group, Astragali Radix and Angelicae Sinensis Radix 1:2 group, Astragali Radix and Angelicae Sinensis Radix 1:5 group, Astragali Radix and Angelicae Sinensis Radix 1:1 group and Astragali Radix and Angelicae Sinensis Radix 5:1 group were lower, and the effects of Astragali Radix and Angelicae Sinensis Radix 1:1 ratio were strongest. The effects on intimal hyperplasia in Astragali Radix group and Astragali Radix and Angelicae Sinensis 2:1 group had no significant differences compared with model group. Conclusion Astragali Radix and Angelicae Sinensis can inhibit vascular intimal hyperplasia in a certain compatibility ratios, and the effects of Astragali Radix and Angelicae Sinensis Radix 1:1 on intimal hyperplasia are the best.
6.Adjustment of MCP-1 to migration and functional activity on macrophages in uterine of inflammatory mice induced by LPS
Huifang DING ; Yuling ZHANG ; Hao LU ; Ruiping ZHANG ; Zhiying QIAN ; Shunli ZHANG
Chinese Journal of Immunology 2017;33(4):507-510
Objective:To explore the adjustment factors to the migration and functional activity on macrophages in the uterine of inflammatory mice induced by LPS.Methods:150 Kunming female mouse were divided into control group (group A),LPS model group (group B),MCP-1 blocking-up group (group C),the mice uterines were extracted separately at hour 1,3,6,12,24.The number of CD14+ macrophages and the expression of CD14 macrophages were detected by Immunohistochemistry,ELISA detects the expression of TNF-α and MCP-1.Results:①Compared with group A,the number of CD14+ macrophages and the expression of CD14 in endometrium,myometrium,perimrtrium of group B were highly significantly increased (P<0.01) at every time points,the endometrium,myometrium of group C were closely to normal level at 1,3,6 h;compared with group B,the number of CD14+ macrophages and the expression of CD14 were highly significantly decreased(P<0.01)at 1,3,6 h of perimrtrium of group C and every time points of endometrium,myometrium of group C.②Compared with group A,the content of TNF-α and MCP-1 were highly significantly increased (P<0.01) at every time points of group B and 12,24 h of group C;compared with group B,the content of TNF-α and MCP-1 of group C were highly significantly decreased(P<0.01) at every time points.Conclusion:The migration of macrophages and the expression of CD14 and TNF-α in the uterine of inflammatory mice induced by LPS were regulated by MCP-1.
7.Effects of TLR/STAT pathway on the proliferation of mesangial cell induced by HMGB1
Lijuan TANG ; Jun HAO ; Ning CHEN ; Huifang GUO ; Qingjuan LIU ; Shuxia LIU
Chinese Journal of Immunology 2001;0(07):-
Objective:To investigate the effects of TLR/STAT pathway in the proliferation of mesangial cell induced by HMGB1.Methods:Human mesangial cells were inoculated in the dose of 1?104 ml-1.After 24 h,cells were cultured with standard medium as control group or with medium supplement with 10 ?g/L human recombinant protein HMGB1 as trial group in vitro.Then the cells were collected in 6,12 and 24 h respectively,as well as control group cells.Immunocytochemical staining was adopted to examine the expressions of PCNA proteins on mesangial cells in different groups.Immunocytochemical staining and FCM were performed to detect the changes of TLR2 protein expression.STAT1 and STAT3 mRNA were examined by RT-PCR technique.Results:Immunocytochemical staining indicated that the mesangial cells could multiply after they were induced by human recombinant protein HMGB1.Immunocytochemical staining showed that the level of TLR2 protein in trial groups were higher than those in control groups.FCM indicated that HMGB1 could significantly up-regulate the expression of TLR2 protein time-dependently.The STAT1 and STAT3 mRNA in HMGB1 groups were higher than those in control groups.The expression of TLR2 protein was positively correlated with those of STAT1 and STAT mRNA respectively.The positive rate of PCNA was remarkably correlated with the expression of STAT1 and STAT3 mRNA.Conclusion:HMGB1 could activate STAT1/STAT3 through combining with its cell-surface receptor TLR2,which may play an important role in promoting the proliferation of mesangial cells and then damaging the renal of lupus nephritis.
8.Effect of NF-κB signal pathway in murine lupus nephritis
Xiaojuan FENG ; Shuxia LIU ; Yujun ZHANG ; Huifang GUO ; Jun HAO ; Ning CHEN ; Lijuan TANG ; Qingjuan LIU ; Haijiang WU
Chinese Journal of Immunology 2010;26(2):169-173,177
Objective:To investigate the expression and mechanism of NF-κB signal pathway in murine lupus nephritis.Methods:The BXSB mice as well as C57BL/6 of 16 weeks were used.Transmission electron microscope and PAS were used to detect the pathological change of renal tissue.RT-PCR and ELISA were used to detect the expression of HMGB1 mRNA and protein.The expression of HMGB1,p- NF-κB,RAGE,IκB and PCNA protein was detected by immunohistochemical stain,FCM and Western blot.Results:The level of BUN in serum and Micro-albumin in urine of BXSB mice was higher than that in C57BL/6 mice.The expression of HMGB1 mRNA and HMGB1 protein level in peripheral blood increased significantly in BXSB group.Compared with those in control group,electron microscopy and PAS revealed the thickness of glomerular basement membrane(GBM),fusion of foot processes partly of epithelial dell and subepithelial electron-dense deposits in the renal tissue of BXSBA mice.Compared with that of control group,expression of PCNA was higher in glomeruli of BXSB mouse.HMGB1 protein over-expression localized in cytoplasm and extracellular milieu,especially in proliferative glomeruli in BXSB group,while the HMGB1 protein primarily confined to the nuclear of tubule in control group.In BXSB group,the expression of p-NF-κB and RAGE increased,while the expression of IκB decreased.There were positive correlation between the expression of HMGB1,RAGE and p-NF-κB protein (r=0.833,0.621,0.848,P<0.01),while the expression of p-NF-κB protein negatively correlated with that of IκB.Conclusion:HMGB1 could activate NF-κB through combining with its receptor-RAGE,induce the form of proliferative glomerulonephritis by promoting the proliferation of inherent cell of glomeruli,which may play an important role in the murine lupus nephritis.
9.The effect of HMGB1 on the renal injure of systemic lupus erythematosus mediated in part via the TLR4 pathway
Shuxia LIU ; Jun HAO ; Huifang GUO ; Yujun ZHANG ; Qingjuan LIU ; Lijuan TANG ; Ning CHEN ; Haifiang WU ; Huijun DUAN
Chinese Journal of Microbiology and Immunology 2008;28(12):1079-1083
Objective To investigate the relationship between the effect of high mobility group protein box 1 (HMGBI) on the renal injure of systemic lupus erythematosus (SLE) and the expression of Toll-like receptor 4(TLR4). Methods The level of HMGB1, MMP-2 and TIMP-2 in serum from 16 pa-tients with SLE, 18 patients with lupus nephritis(LN) and 12 healthy people were measured by ELISA. The fresh peripheral blood mononuelear cell (PBMC) were isolated and the total RNA was extracted. Then the mRNA expression of HMGB1 was amplified by RT-PCR. Flow cytometry analysis was performed to study cell surface markers and the expression of TLR4. Results RT-PCR and ELISA results showed that the expres-sions of mRNA and level of HMGB1 protein in serum were higher in patients with LN than those in SLE and healthy people. The expression of TLR4 in CD14+ monecytes of patients with LN was higher than that with SLE and healthy people, while there were no significance in CD3+ T cells among LN, SLE and healthy peo-ple. The expressions of MMP-2 and TIMP-2 in serum of LN was lower than that in SLE and healthy people, at the same time the ratio of MMP-2/TIMP-2 decreased in LN group. HMGB1 mRNA and CD14+/TLR4+ was negatively correlated with the ratio of MMP-2/TIMP-2, and the level of HMGB1 in serum was positively correlated with proteinuria, while negatively correlated with the ratio of MMP-2/TIMP-2 in LN. Conclusion HMGB1 is one of the important cytokine in the pathogenesis of lupus nephritis. HMGBI might play a role in proteinuria of lupus nephritis in part via TLR4 pathway to activate monocytes and decrease the expression of MMP-2/TIMP-2.
10.Subjective global assessment predicts the prognosis of patients with hospital-acquired acute kidney injury
Ying ZHOU ; Huifang GU ; Qionghong XIE ; Zhongye XU ; Shuai MA ; Huaizhou YOU ; Dingwei KUANG ; Yong GU ; Chuanming HAO ; Shantan LIN ; Feng DING
Chinese Journal of Nephrology 2011;27(8):567-571
Objective To elucidate the malnutrition in patients with hospital-acquired acute kidney injury(AKI), and to examine the association betweensubjective global assessment (SGA) and prognosis.Methods Adult patients with hospital-acquired AKI were prospectively enrolled in this cohort study.Nutritional evaluations, including SGA, anthropometric and serum nutritional markers were conducted at enrollment.Overall survival at 90 days among different SGA scores was analyzed using Kaplan-Meier methods, and differences were tested using the log-rank test.The Cox model was used to analyze the relationship between SGA scores and all-cause mortality after adjusting for confounders.Results A total of 170 patients were enrolled.The prevalence of moderate malnutrition(SGA B) and severe malnutrition(SGA C) was 51.8% and 22.9% respectively, while patients with normal nutrition(SGA A) accounted for 25.3%.After 90 days follow-up, all-cause mortality was 9.8% in SGA A group, 34.9% in SGA B group and 56.8%inSGACgrouprespectively. Afteradjustingforage,sex,dialysis,ventilation, hemoglobin, platelets and bilirubin, the hazard ratio(HR) of 90 days all-cause mortality was 4.0(95% CI 1.42-11.22, P=0.008) in malnutrition group (SGA B group and SGA C group) compared with SGA A group.The Kaplan-Meier curve also revealed that the worse the SGA score was, the lower the cumulative survival became (P<0.01).Conclusion SGA score is an independent risk factor for all-cause mortality within 90 days in patients with hospital-acquired acute kidney injury.