1.Phage display of human CD137 extra-cellular domain and its bioactivity detect
Na LI ; Lining ZHANG ; Qun WANG ; Chenghu LIU ; Xingtian LIU
Chinese Journal of Immunology 1986;0(04):-
Objective:To display human CD137 extra-cellular domain on phage using phage display technique and detect its antigenicity and bioactivity.Methods:CD137 extra-cellular gene was amplified from CIS plasmid using PCR and then clone it into pComb3HSS.The recombinant pComb3H-CD137 was transfected into competent bacteria XL1-Blue, then the transfected bacteria was co-infected with help phage VCSM13 and CD137 extra-cellular domain was displayed on the phage surface. The antigenicity and bioactivity of CD137 displayed on phage was detected by ELISA and inhibition assay of lymphocyte proliferation(MTT).Results:CD137 PCR products was 525 bp, the recombined plasmid was named pComb3H-CD137, CD137 was displayed on phage. ELISA detection showed that phage displayed CD137 possesses antigenicity of human CD137. MTT inhibition assay showed that lymphocytes proliferation stimulated by PHA and CD137 could be inhibited by phage displayed CD137, indicating that it has bioactivity of nature CD137.Conclusion:The studies demonstrate that phage display system can display human CD137 extra-cellular domain homodimer with antigenicity and bioactivity.
2.Inhibitory effects and tocolysis role of fructus polygoni orientalis on macrophages in uterine of abortion mice induced by LPS
Zhiying QIAN ; Yajie LIU ; Yanpin CUI ; Lining XIE ; Shunli ZHANG
Chinese Journal of Immunology 2014;(6):763-767
Objective:To explore the immunosuppression effects and tocolysis of fructus polygoni orientalis on abortion mice induced by LPS.Methods:Mice of Kunming (55 mice) were divided into control group (group A ,10 mice) and experimental group. Experimental group were divided into group B ( intraperitoneal injection LPS ) , group C ( ingestion fructus polygoni orientalis ) , and group D (ingestion fructus polygoni orientalis and intraperitoneal injection LPS ),each group of 15 mice.Then the pregnancy results were observed ,the positive of α-NAE+and the varity of CD 14+and CD204+macrophages ,TNF-αin the uteri were identified by enzyme-histochemistry ,immunohistochemistry and ELISA.Results:The abortion rate and the embryo resorbing rate were all 100%( P<0.01 ) in group B.But there was the decreased abortion rate of 13.33% in group D .The embryo resorbing rate decreased to 10.39%.The number and positive cell area of α-NAE+and CD14+macrophages in the uteri of gestation mice of group B was greatly increased comparing with group A ( P<0.01 ) .These effects outside of myometrium of group D were remarkably increased comparing with group A (P<0.01),but there was no distinct difference in the inside of myometrium and function layer.The number and positive cell area of CD204+macrophages in group C and D was greatly increased comparing with group A and B ( P<0.01 ) .The TNF-αcontents in the uteri of mice in group B were greatly increased comparing with group A (P<0.01),but the positive cell area of CD14,CD204 were close to normal levels in group D.Conclusion:The effect of miscarriage induced by LPS is antagonized by fructus polygoni orientalis through inhibiting the phenotype ,activity and function characteristics of macrophages in the uteri of gravidity mice.
3.Effect and clinical significance of Tacrolimus on serum interleukin-2(IL-2),soluble interleukin-2 receptor(sIL-2R) expression in refractory nephrotic syndrome
Lining MIAO ; Yanan ZHANG ; Shengmao LIU ; Yingchun CUI
Chinese Journal of Immunology 1985;0(06):-
Objective:To investigate the effect of Tacroliums(FK506) on the significance of IL-2 and sIL-2R expression in refractory nephrotic syndrome. Methods: The expression of IL-2 and sIL-2R were evaluated by enzyme-linked immunosorbent assay (ELISA) on both pretreatment and post-treatment groups, and supervise the changes of 24hours urinary protein,serum albumin and lipid. Results:Serum IL-2 and sIL-2R level were higher in pre-treatment group than in normal control group(P
4.The influence of methyl-prednisone on extracellular matrix in rats with immune commplexes nephritis
Guangdong SUN ; Shujun LIU ; Lining MIAO ; Hua GU ; Ping LUO
Chinese Journal of Immunology 2000;0(08):-
Objective:To observe the theraputic effect and changes of glomerular extracellular matrix components(ECM) and PAI-1 and their relationships after cure with methyl-prednisone on immune complexes nephritis rats.Methods:Immune complexes nephritis rats model were induced with C-BSA.Levels of FN,LN in different treatment groups were analyzed by ELISA, level of PAI-1 in rats renal tissue was determined by color developing substrate,was theraputic effect of methyl-prednisone with 24 hours volumes of urine protein.Results:The levels of PAI-1,FN and LN of model groups were significantly higher than those of normal and control groups,and PAI-1 was significantly correlated with LN and FN,Glomerular mesangial matrix proliferated slightly and moderately;The levels of FN,LN and PAI-1 decreased signifcantly and glomerular mesangial matrix proliferation lessen differently after cure of methyl-prednisone for 1 and 2 weeks,but not reaching the level of normal groups;24 hours volumes of urine protein decrease significantly in treatment groups.Conclusion:The ECM accumulation correlate with PAI-1 increase in immune complexes rats,methyl-prednisone may affect ECM accumulation by interfering with PA/PAI-1 system to reach a treatment purpose.
5.Effect of Entecavir Combined with Dahuangzhechong Pills on Hepatic Fibrosis in Patients with Chronic Hepatitis B
Chunping LI ; Lining DENG ; Hongbo HOU ; Xiurong LIU
China Pharmacy 2005;0(14):-
0.05).After treatment,serum hyaluronic acid(HA),laminin(LN),type Ⅲ precollagen,type Ⅳ collagen peptide decreased more significantly in the treatment group than in the control group(P
6.Effect of Liuweidihuang Granula Cooperating with Flixotide Aerosol on IFN-? and IL-4 of Asthma Rat Models
Han LIU ; Lining WANG ; Xiaoqi HUANG ; Liying LIANG ;
China Journal of Traditional Chinese Medicine and Pharmacy 2006;0(09):-
Objective: To observe the effect of representative formula of nourishing yin and invigorating the kidney(Liuweidihuang Granula) cooperating with Flixotide aerosol inhalation on the allergic airway inflammation of asthma rat models.Methods: Wistar rats were randomly divided into six groups.Except the norma1 control group,all the other 5 groups were sensitized by injecting into the abdominal cavity of the ovalbumin(OVA) together with aluminum hydroxide as adjuvants,2 weeks later followed by aerosolized OVA challenge to prepare the asthma models.From the eighth day of the experiment,in medication administration teams, rats were sprayed with Flixotide aerosol and given intragastric administration with Liuweidihuang Granula.The normal control group and the asthma model group were drenched with normal saline.Results: There was no difference between the groups given Liuweidihuang Granula and different dosages of Flixotide aerosol but the amount of IFN-? and IFN-?/IL-4 in BALF in the Liuweidihuang Granula cooperating with Flixotide aerosol group were higher than that in the model group(P
7.The clinical significance of microproteinuria and the correlation between microproteinuria and renal pathological changes in patients with systemic lupus erythematosus
Yu WANG ; Linlin LIU ; Yanqiu LI ; Lining WANG
Chinese Journal of Rheumatology 2001;0(04):-
Objective To explore the clinical significance of urinary ?1-microglobulin (?1-MG), ?2-micro-globulin (?2-MG), albumin (MA), immunoglobulin G (IGU) and transferritin (TRU), and investigate the correlation between microproteinuria and renal pathological changes in patients with systemic lupus erythematosus (SLE). Methods According to the presence of nephritis, 47 SLE patients who had renal biopsy were divided into two groups: 30 patients with clinical lupus nephritis (OLN), and 17 patient with silent lupus nephritis (SLN). The urinary ?1-MG, ?2-MG, MA, IGU and TRU were measured in SLE patients and compared with healthy control group. The level of microproteinuria and renal biopsy pathological changes in SLE patients were studied. Results ①The levels of urinary ?1-MG, ?2-MG, MA, IGU and TRU had statistical differences between SLE group and healthy control group, OLN group and SLN group, SLN group and healthy control group respectively. ②The levels of urinary MA、TRU、 IGU were significantly higher in WHO class Ⅳ than in class Ⅱ LN, the level of urinary MA was markedly increased in class Ⅳ than in class Ⅲ LN and ?1-MG was significantly different between class Ⅳ and Ⅴ LN. In classⅤ LN, the urinary MA was obviously increased compared with class Ⅱ LN. Conclusion ①The urinary ?1-MG, ?2-MG, MA, IGU and TRU reflects the severity of renal damage in SLE patients and may be used as sensitive markers for early lupus nephritis. ②The measurement of microproteinuria indicates that the difference in severity of renal damage exists among different WHO classes of LN and urinary MA and ?1-MG may be sensitive indicators.
8.Glomerular C4d Deposition Is a Prognostic Factor for IgA Nephropathy
Linlin LIU ; Nan LIU ; Juan WANG ; Linbo ZHU ; Li YAO ; Tianhua XU ; Lining WANG
Journal of China Medical University 2017;46(1):23-27
Objective To investigate the significance of glomerular deposition of C4d in accessing the severity and prognosis of IgA nephropathy. Methods A total of 131 patients were recruited for the study. Immunofluorescence was used to detect the deposition of C4d in renal tissue of pa?tients with IgA nephropathy,and the relationship between C4d deposition and clinical and pathological parameters and renal remission was ana?lyzed. Results Totally 30 patients had glomerular deposition of C4d. Compared with the patients without C4d deposition,the patients with C4d deposition had significantly higher levels of serum creatinine,urinary protein excretion and C4d and higher prevalence of hypertension,but had sig?nificantly decreased levels of glomerular filtration rates. With the histopathological phenotypes segregated by Lee 's classification,the ratios of C4d deposition presented an increase(P=0.005). The patients with C4d deposition had more severe mesangial proliferation,endocapillary hypercellu?larity,segmental glomerulosclerosis and tubular?interstitial injury. The rates of renal remission were significantly lower in IgA nephropathy patients with C4d deposition than those without C4d deposition(P<0.001). Conclusion IgA nephropathy patients with C4d deposition have more se?vere clinical and pathological manifestations and lower rate of renal remission. Glomerular C4d deposition is expected to be an important pathologi?cal prognostic factor for predicting the prognosis of IgA nephropathy.
9.Effect of sevoflurane postconditioning on activities of Na+-K+-ATPase and Ca2+ -Mg2+ -ATPase during myocardial ischemia-reperfusion in rats
Yue LIU ; Jin ZHANG ; Ya LIU ; Jianjun REN ; Lining HUANG ; Xuze LI ; Zhenming DONG
Chinese Journal of Anesthesiology 2010;30(10):1179-1181
Objective To investigate the effect of sevoflurane postconditioning on the activities of Na+ -K+ -ATPase and Ca2+ -Mg2+ -ATPase during myocardial ischemia-reperfusion (I/R) in rats and the possible mechanism. Methods Forty-five healthy male Wistar rats weighing 250-280 g were randomly divided into 3 groups ( n = 15 each): sham operation group (group S), I/R group and sevoflurane postconditioning group (group Spo). Myocardial I/R was induced by occlusion of anterior descending branch of left coronary artery for 30 min followed by 120 min of reperfusion. In group S the anterior descending branch was only exposed but not ligated. Group Spo received 5 min inhlation of 2.5% sevoflurane 1 min before reperfusion. The myocardial tissues were taken at 2 h of reperfusion for determination of infarct size and activities of Na+ -K+ -ATPase and Ca2 * -Mg2 * -ATPase. Results The infarct size was significantly larger and the activities of Na+ -K+ -ATPase and Ca2+ -Mg2+ -ATPase were signifi cantly lower in group I/R than in group S ( P < 0.05). The infarct size was significantly smaller and the activities of Na+ -K+ -ATPase and Ca2+ -Mg2+ -ATPase were significantly higher in group Spo than in group I/R (P < 0.05 ). Conclusion Sevoflurane postconditioning can reduce myocardial I/R injury through increasing the activities of Na+ -K+ -ATPase and Ca2+ -Mg2+ -ATPase.
10.Role of RhoA/ROCK2 signaling pathway in propofol-induced neuroapoptosis in hippocampus of newborn rats
Xuze LI ; Xuefang LIU ; Lining HUANG ; Lijun BO ; Ya LIU ; Rongtian KANG ; Zhenming DONG
Chinese Journal of Anesthesiology 2017;37(7):817-820
Objective To evaluate the role of Ras homolog family member A (RhoA)/Rho-associated coiled-coil containing protein kinase 2 (ROCK2) signaling pathway in propofol-induced neuroapoptosis in the hippocampus of newborn rats.Methods Experiment Ⅰ Primarily cultured hippocampal neurons were seeded in 6-well culture plates at a density of 1×106 cells/ml and divided into 2 groups (n=6 each) using a random number table:solvent control group (C group) and propofol group (P group).Propofol was added with the final concentration of 60 μg/ml in group P.Dimethyl sulfoxide was added with the final concentration of 0.04% in group C.The expression of RhoA and ROCK2 in hippocampal neurons was measured by Western blot at 24 h of incubation.Experiment Ⅱ Primarily cultured hippocampal neurons were seeded in 6-well culture plates at a density of 1 × 106 cells/ml and divided into 3 groups (n =6 each) using a random number table:solvent control group (C group),propofol group (P group) and propofol plus specific RhoA/ROCK2 signaling pathway blocker Y27632 group (P+Y group).Propofol was added with the final concentration of 60 μg/ml in group P.Propofol at the final concentration of 60 μg/ml and Y27632 at the final concentration of 10 μmol/L were added in group P+Y.Dimethyl sulfoxide was added with the final concentrauon of 0.04% in group C.At 24 h of incubation,the neuroapoptosis in hippocampi was detected by flow cytometry,and the expression of activated caspase-3 in hippocampal neurons was measured by Western blot.The apoptotic rate was calculated.Results Experiment Ⅰ Compared with group C,the expression of RhoA and ROCK2 in hippocampal neurons was significantly up-regulated in group P (P<0.05).Experiment Ⅱ Compared with group C,the apoptotic rate of hippocampal neurons was significantly increased,and the expression of activated caspase-3 was up-regulated in P and P+Y groups (P<0.05 or 0.01).Compared with group P,the apoptotic rate of hippocampal neurons was significantly decreased,and the expression of activated caspase-3 was down-rcgulatcd in group P + Y (P< 0.05).Conclusion Activation of RhoA/ROCK2 signaling pathway is involved in propofol-induced neuroapoptosis in hippocampi of newborn rats.