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Objective:To evaluate the therapeutic effects of Tolvaptan on refractory heart failure in patients aged 75 years and older.Methods:This was a randomized controlled trial.A total of 68 patients with refractory heart failure aged 75 years and older were divided into the control group(n=38)and the experimental group(n=30)by randomly generated numbers.Patients in the control group were given levosimendan and recombinant human brain natriuretic peptide intravenously plus routine treatments such as diuresis and electrolyte correction.In the experimental group, 30 patients were given a single dose of 15 mg Tolvaptan per day in addition to what was received by the control group.The effects on heart failure were compared between the two groups 1 week after treatment.Changes in rehospitalization rate, emergency intervention frequency and mortality rate were recorded after a 3-month follow-up.Results:Clinical symptoms of heart failure were alleviated in both the experimental and control groups after treatment.Improvements in 24-h urine volume, body weight and 6-minute walking distance were more significant in the experimental group than in the control group after treatment[(1 470.5±200.6)ml vs.(972.5±201.7)ml, (-6.4±2.1)kg vs.(-2.8±1.9)kg, (189.3±13.7)m vs.(151.3±12.5)m, P<0.05]. Changes in serum sodium levels and improvement of LVEF were greater and reduction of N-terminal B-type brain natriuretic peptide(NT-proBNP)levels was more significant in the experimental group than in the control group after treatment[(5.2±2.1)μmol/L vs.(-1.1±2.4)μmol/L, (10.1±4.1)% vs.(7.0±4.0)%, (-6 670±1 815.7)ng/L vs.(-5 025.3±1 876.7)ng/L, P<0.05]. There was no significant difference in the incidence of adverse reactions between the two groups( P>0.05). The experimental group had shorter hospital stays, while the rehospitalization rate, emergency intervention times and mortality had no significant difference between the two groups during the follow-up period( P>0.05). Conclusions:Addition of Tolvaptan to treatment can increase urine volume, improve cardiac function, correct hyponatremia and shorten the length of hospitalization in refractory heart failure patients aged 75 years and older with good safety and has no significant impact on renal function.
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Objective:To study the change of related molecular about apoptosis we reduce the expression of CD59 on acute T lymphocyte Jurkat cell lines .Methods: RNA interference (RNAi) was used to reduce the expression of CD59 gene by lentivirus,confocal was applyed to observe the transfection efficiency and the location of CD59 molecular then Real-time-PCR and Western blot were used to select the most effective group to do the rest experiment;Western blot was used to detect the change of expression about Bcl-2,Bax,Caspase-3 and Survivin;ELISA was used to investigate the expression of IL-3 and TNF-β.Results: Confocal observed each group′s transfection efficiency over 90%,CD59 molecules were mainly located in cell membrane;Real-time-PCR and Western blot showed group A had the best down-regulation efficiency;we defined RNAi-CD59-A as experimental group for subsequent experiments named RNAi-CD59;the experimental group can enhance the expression of Bax,caspase-3 (P<0.05),inhibit the expression of Bcl-2 and Survivin(P<0.05);ELISA showed that the expression of IL-3 in the down-regulation group increase(P<0.05),the expression of TNF-β decrease (P<0.05).Conclusion: Down-regulation CD59 can promote the expression of apoptosis molecular in acute leukemia Jurkat cell lines restrain the expression of proliferation molecular.
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Objective:To explore the proportions of Th17 cells in the peripheral blood and levels of IL-17,IL-23 in the serum of patients with Graves′disease ( GD ) and their clinical significance.Methods: We studied 29 patients with GD ( GD group ) , and reevaluated the GD group after therapy ( euthyroid GD group ).29 gender-and age-matched volunteers were selected as the normal control ( NC group).The proportions of Th17 cells were investigated by flow cytometry.The levels of IL-23,IL-17 in the serums were measured by ELISA.The levels of FT3,FT4,TSH were determined by ECLIA and the levels of TrAb were tested by RRA.Results:There were no significant difference among 3 study groups in sex and age match ( F=0.0075 , P>0.05;χ2=0.4213 , P>0.05 ).The proportions of Th17 cells and the levels of IL-17 , IL-23 were increased in the GD and euthyroid GD patients compared with the control group (respectively,P0.05).Correlation analysis revealed that the proportions of Th17 cells ,and the levels of IL-17,IL-23 were positively correlated with the levels of FT3,FT4,TrAb(r=0.588 2,0.337 2,0.371 0;0.549 6,0.287 5,0.342 7;0.361 0,0.420 8, 0.330 8;P<0.05 ,for all parameters ) ,and were negatively correlated with the levels of TSH ( r=-0.319 7 ,-0.472 8 ,-0.428 2;P<0.05,for all parameters).Conclusion:Th17 cells and their related cytokines IL-17,IL-23 are highly expressed in the serum of patients with GD.Th17 cells and their relative cytokines have certain relevance with 4 thyroid function parameters of the patients with GD , which can be used as biological markers for GD.
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Objective:To establish T lineage leukemia Jurkat cell mice model of over expression of C-terminal Src kinase binding protein( Cbp ) and Cbp palmitoylation and to research the effect of Cbp and Cbp palmitoylation to proliferation of Jurkat cell.Methods:Virus transfected cell of neg-EGFP,Cbp-EGFP and Cbp-m-EGFP were used in mice model.24 female BALB/c-nu mice were randomly divided into blank control group,empty virus control group,over expression of CBP group and Cbp palmitoylation group, 6 mice in each group.The nude mice were weighed in 0,1,2,3,4,5 weeks.The amount of white blood cell in peripheral blood were counted in 0, 1, 2, 3, 4, 5 weeks.The proliferation of Jurkat cell in peripheral blood of mice were observed by laser confocal microscope.The pathological changes of liver were observed using HE staining.The proliferation of Jurkat cell in the bone marrow and peripheral blood of mice were detected with flow cytometry.Results:The weight of mice in over expression of Cbp group was less than that in blank control group,but higher than that in empty virus control group and Cbp palmitoylation group.The weight of mice in Cbp palmitoylation group was less than that in blank control group,empty virus control group and over expression of Cbp group.The amount of white blood cell in peripheral blood and proliferation of Jurkat cell in liver, bone marrow and peripheral blood of mice in over expression of Cbp group was higher than that in blank control group, but less than that in empty virus control group and Cbp palmitoylation group.The amount of white blood cell in peripheral blood and proliferation of Jurkat cell in liver, bone marrow and peripheral blood of mice in Cbp palmitoylation group was higher than that in blank control group,empty virus control group and over ex-pression of Cbp group.Conclusion:Over expression of Cbp and Cbp palmitoylation in T lineage leukemia Jurkat cell mice model was established.Over expression of Cbp has inhibitory effect on the proliferation of Jurkat cell.Cbp palmitoylation has promotable effect on the proliferation of Jurkat cell.
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BACKGROUND:The linkage and synergistic effect of adaptor proteins can effectively regulate signal transduction of T cel s, which can form a limit or amplification cascade to realize the complex immune function of T cel s. C-terminal Src kinase (Csk)-binding protein (Cbp) is an adaptor protein, which mainly exert the negative feedback regulation of Src kinase activity. This negative feedback effect depends on Y317 of Cbp, which may be involved in the SH2 domain of Csk. OBJECTIVE:To explore the effects of high expression of Cbp on ultrastructure and related biological function of Jurkat cel s. METHODS:The virus particles were constructed with expressing enhanced green fluorescent protein (EGFP) only and Cbp-EGFP fusion protein to transfect Jurkat cel s. There were untransfected group (Jurkat group), negative control group (transfected with expression of EGFP virus only), and Cbp group (transfected with Cbp-EGFP virus). RESULTS AND CONCLUSION:Confocal microscope showed that cel transfection efficiency was more than 95%and Cbp was located on the cel membrane. Optical microscope showed after transfection with Cbp-EGFP virus, more Jurkat cel s shrunk, with poor size uniformity. Apoptosis detection showed that after transfection with Cbp-EGFP virus, the number of apoptotic and necrotic cel s was greatly increased. Cbp mRNA expression was increased, Csk expression was decreased obviously and lymphocyte-specific protein tyrosine kinase expression was increased. So, in Jurkat cel s, the high expression of Cbp can decrease the uniformity of cel s and increase the necrosis cel s, thus inhibiting the signal transduction.
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Objective:To investigate the effects of chitin on atopic dermatitis in an OVA induced AD murine model.Methods:Twenty-eight BALB/c mice were randomly divided into three groups:the normal control group (N)(8),the chitin group(E) (10) and the AD model group(M)(10).The murine model of atopic dermatitis was established through intraperitoneal injection of OVA followed by repeated epicutaneous application of OVA on mice back skin( AD model group).During the set up of AD murine model,mice of the chitin group were given intragastric gavage of 3 mg/d for 4 weeks.At the end of the experiment, the mice were sacrificed and skin lesions were biopsied for histological study.HE and O-toluidine stained paraffin sections were observed under microscope.The spleen cells were cultured and challenged with OVA and chitin,respectively,the supernatant was obtained for cytokine determination.Serum levels of total and OVA-specific IgE and total IgG2a were determined with ELISA.Results:Chitin significantly inhibited skin inflammation induced by OVA.Compared with the AD model group,the thickness of the epidermis and dermis in the chitin group were obviously decreased.The numbers of dermal infiltrated inflammatory cells,eosinophils and mast cells were significantly decreased in the chitin group compared with the AD model group ( P<0.05-0.001 ).The serum level of total IgE and OVA-specific IgE were significantly lower in the chitin group than in the AD model group(P<0.05-0.001),while the serum level of IgG2a in the chitin group was significantly higher than that of the AD model group( P<0.001).The cultured spleen cells of the chitin group produced significantly higher levels of IL-12 and IFN-γ,but lower level of IL-4 compared with those of the AD model group after OVA challenge (P<0.05).Conclusion:Chitin can inhibit the inflammation and decease the seum level of IgE in the murine AD model.The antiallergic effect of chitin might be associated with the induced production of Th1 type cytokines by mice spleen cells.
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Objective To explore the optimal immunohistochemical (IHC)diagnostic index for supraclavicular lymph node me-tastasis in lung cancer.Methods In the premise of conforming to the evidence-based medicine,the biopsy of lymph node in 135 pa-tients with supraclavicular lymph node metastasis was performed and 7 antibodies were selected to be detected.135 cases were di-vided into two classes:class A (110 cases)and B (25 cases).The class A included 14 cases of primary lesion inside the lung by the histological analysis,43 cases of primary lesion inside the lung by the cytological detection and 53.cases of primary lesion inside the lung by the radiographic analysis.The class B included 25 cases of lesion outside the lung by the histological analysis.The detecting antibodies included CK7,CK20,EMA,CEA,TTF-1,SPB and vimentin.Results The sensitivity and specificity of single antibody for deducing the primary lesion inside the lung were 90.0% and 56.0% for CK7,98.0% and 40.0% for CK20,90.9% and 4.0%for EMA,80.9% and 36.0% for CEA,62.7% and 100.0% for TTF-1,65.6% and 100.0% for SPB,and 60.9% and 60.0% for vimentin.The quadruple antibodies (CK7,CK20,TTF-1 and SPB)showed the highest sensitivity(85.0%)and highest specificity (100.0%),which made the relatively high detection rate (80.0%)of lung cancer in clinic.Conclusion Analyzing and researching the IMC results is of great significance to guide the surgical pathological practice based on the evidence-based medicine.
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Objective:To explore the role of cytokines in the pathogenesis of Graves′disease(GD),by detecting the levels of IFN-γ,IL-6,IL-17 and TGF-β1 in GD patients who were newly diagnosed.Methods:A total of 23 patients with new onset GD and 23 gender-and age-matched healthy controls were examined.The levels of serum IFN-γ, IL-6, IL-17 and TGF-β1 were measured by ELISA,FT3,FT4 and TSH levels were determined by ECL IA;TrAb levels were tested by RRA.Results: There were no significant difference among GD and NC group in sex and age match ( t=0.334 8 ,P>0.05;χ2=0.410 7 ,P>0.05 ).The levels of serum IFN-γ,IL-6,IL-17 and TGF-β1 in the GD group were significantly higher than the control group ( P<0.05 ) .Correlation analysis revealed that IFN-γ,IL-6,IL-17 and TGF-β1 were positively correlated with FT3,FT4(r=0.324 6,0.453 2,0.431 0,0.463 8;0.413 2,0.441 5, 0.436 2,0.467 1;P<0.05 ).Conclusion: IFN-γ, IL-6, IL-17 and TGF-β1 are highly expressed in the newly diagnosed GD patients.They play an important role in the pathogenesis of GD ,and provide helpful evaluation indices of immune dysfunction to Graves disease.
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Objective:To explore the vailation of serum sP-selectin,sE-selectin,levels in the patients of pre-diabetic,simple type 2 diabetes mellitus (T2DM) and T2DM with coronary heartdisease(CAD).To investigate the possible mechanism that sP-selectin, sE-selectin accelerate type 2 diabetes mellitus.Methods: Level of serum sP-selectin, sE-selectin was assayed by ELISA in type 2 diabetes with or without coronary heart disease (32 and 34 cases respectively),pre-diabetic (32 cases) and control group(32 cases). Meanwhile BMI,BP,FBS,FINS,TG,CHO,HDL-C,LDL-C,HbA1C were determined in all cases as well as in control group.Results:The serum levels of sP-selectin and sE-selectin in pre-diabetic,type 2 diabetes mellitus with or without coronary heart disease groups were significantly higher than the control group ( P<0.01 ).There was significant positive correlation between serum levels of sP-selectin,sE-selectin and these items including FBG ,FINS,TG,HbA1C,HOMA-IR(P<0.01);but sE-selectin was negatively correlated with HDL-C (P<0.05).Conclusion:sP-selectin,sE-selectin,are risk factors in the initiation and progression of pre-diabetic,type 2 diabetes with or without coronary heart disease;sP-selectin and sE-selectin possibly accelerate type 2 diabetes by inducing insulin resist-ance.
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Objective:To investigate the function of CD 59 in LAT induced T lymphocytes'proliferation and activation.Methods:Transfected LAT-GFP recombinant lentiviral vectors into Jurkat cells and established a fusion-protein stable express cell line ( Jurkat-GFP ).Junket-GFP cells were transfected with pSUPER-siCD59 plasmids by electroporetion or stimulated by anti-CD59 antibody.The cellular locations of CD 59 and LAT were observed under fluorescence microscope with the immunofluorescence cytochem -istry.The cells proliferation were measured by MTT assay.Furthermore,Western blot was used to detect the total and phosphorylation levels of several down-stream proteins after T cell activated .Results: Jurkat-GFP cells successfully transfected with pSUPER-siCD59 plasmids showed lower fluorescence staining.CD59 and LAT distributed uniformly on the cell surface before stimulated with anti-CD59 antibody and formed clusters once upon stimulation.Jurkat-GFP cells stimulated with anti-CD59 antibody showed a higher level of pro-liferation and protein phosphorylation ,compared with the others.Conclusion:CD59 contributed to LAT induced signaling transduction of T lymphocytes ,and stimulated CD59 molecule partly promoted T cell activation.
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Objective To investigate the value of combination detection of CA15-3,CEA,COX-2 on early diagnosis of breast carcinoma.Methods 68 breast cancer patients,61 benign breast disease patients and 68 healthies were selected.Serum levels of COX-2 were detected by ELISA technique,CA15-3 and CEA levels were detected by electroche-miluminescence technique.Results Serum levels of CA15-3,CEA,COX-2 in breast carcinoma group were (34.67±13.20) U/ml,(7.38±3.87) ng/ml,(43.25±10.87) ng/ml.Their levels were higher than those in benign breast disease group and healthy group,the differences were significant (P ＜ 0.01).In combination detection,the sensitivity was 84.9 ％,and accuracy was 91.2 ％,which were higher than single tumor marker used in breast cancer diagnosis.Conclusion The combined detection of CA15-3,CEA,COX-2 could increase the diagnosis rate of breast carcinoma.
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Based on their inherent relationship,some independent and verifying experiments of medical immunology were intergrated into a series of comprehensive and open experiments,which was embodied in the preparations of antiserum.so as to set up a new experiment teaching system emphasizing the training of student comprehensive capability.Achievements in training students'scientific research capacity,innovative motivation and practical working ability have been obtained and the results could provide valuable experience for innovation and reform of medical immunology experiment teaching model and method.
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Objective To observe the effect of gheocorticoid by oral medication or intrathoracic iniection on the expression of IL-6,IL-8 and sIL-2R in gerum and pleural fluid in patients of tuberculous pleurisy.Methods Twenty tuberculous pleurisy patients were treated with ghcocortieoid by oral medication (oral group)and 20 cases by intrathoracic injection(injection group).ELISA wag employed to detect the levels of IL-6,IL-8 and sIL-2R in serum and pleural fluid before treatment and 3,6 and 9 days after treatment.Results The levels of serum IL-6 and IL-8 decreased and sIL-2R increased obviously in injection group at 3,6 and 9 days after treatment(P＜0.01).Meanwhile the levels of IL-6 and IL-8 in pleural fluid decreased and sIL-2R increased obviously(P＜0.01).The levels of IL-6 and IL-8 were significantly lower and the levels of aIL-2R were significantly higher in injection group than those in oral group(P＜0.01).One month after treatment,the rate of complete absorpion of pleural fluid was higher in injection group(80%)than thatin oral group(45%),the rate of pachynsis pleurae was lower in injection group(10%)than that in oral group(40%),P＜0.05.Conclusions Intrathoracic injection of ghcocorticoid shows stronger suppression of cellular immune function in cavum pleurae and weaker suppression of the general immune sysmm than oral medication.Intrathoracic injection of glucocorticoid can increase the therapeutic effect and decrease side effect in tuberculous pleurisy.
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Objective:To investigate the inhibitiory effect of CD59-siRNA on CD59 gene expression and the growth of xenografted ovarian cancer in vivo,and to explore the role of CD59 in tumor immune escape.Methods:A2780 cells transfected with siRNA plasmid (T group),A2780 cells transfected with blank plasmid (V group) and the wild A2780 cells (C group) were subcutaneously injected into nude mice,respectively.The inhibitiory effect of CD59-siRNA on tumor growth and CD59 expression was evaluated by tumor growth curves and in situ hybridization (ISH) and immunohistochemistry (IHC).Results:The tumor growth curves demonstrated that the growth of cells transfected with siRNA plasmid was significantly inhibited (P<0.05).ISH and IHC showed that the expression of CD59 mRNA and CD59 protein were decreased significantly compared with other groups (P<0.05).Conclusion:Studies of in vivo experiment demonstrate that CD59-siRNA significantly inhibites the expression of CD59,increases the sensitivity of A2780 cells to the complement attack and inhibites the tumor growth.The results may further indicate the role of CD59 in tumor immune escape.
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Aim To investigate the influence and molecular mechanism of C-phycocyanin(CPC) from Spirulina platensis on apoptosis of HeLa cells in vitro.Methods Firstly,the effect of purified CPC on proliferation of HeLa cells in vitro was determined by MTT assay,and then electron microscope was exploited to observe the characteristic apoptotic features of cells treated with CPC.Subsequently,genomic DNA changes of HeLa cells were observed by agarose electrophoresis.Flow cytometric analysis revealed the influence of different concentrations of CPC on cell cycle of HeLa cells.The expressions of apoptosis related genes of CPC treated HeLa cells were determined by immunohistochemistry analysis.In addition,the activities of caspases and the release of cytochrome c from mitochondria into the cytosol were detected.Results Compared with control cells untreated with CPC,a significant decreased in the numbers of HeLa cells in survival treated with CPC and concentration dose effects existed.CPC could induce characteristic apoptotic features including cell shrinkage,membrane blebbing,microvilli loss,chromatin margination and condensation into dense granules or blocks.DNA of HeLa cells treated with CPC showed fragmentation pattern(DNA ladder of oligomers of 180~200 bp) typical for apoptotic cells.HeLa cells treated with different concentrations of CPC demonstrated an increasing percentage of cells in sub-G0/G1 phase.In addition,CPC could promote the expression of pro-apoptotic gene(Fas and ICAM-1);meanwhile,held back the anti-apoptotic gene(Bcl-2) expression,and then facilitated the transduction of tumoral apoptosis signals that resulted in the apoptosis of HeLa cells in vitro.In CPC treated HeLa cells,CPC treatment of HeLa cells also resulted in activation of caspases and release of cytochrome C from mitochondria into the cytosol.Conclusion C-phycocyanin from Spirulina platensis can induce the apoptosis of HeLa cells in vitro.By virtue of the promotion of the apoptosis signals transduction in HeLa,CPC realizes its antitumor activities.
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6).The genes were genes that associated with the regulation of cell differentiation and proliferation,adhesion and signal tranduction,apoptosis,transcription and modulation,and DNA damage and repair that.Conclusion:The differential hybridzation analysis of Atlas cDNA expression arrays can be a useful method for analysing the expression profiles of PBMC genes and studing genes differential expression in SLE and IDDM patients.
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Objective To study chemcial constituents in the canes of Marsdenia tenacissima.Methods Chemical constituents were isolated and purified by silica gel and ODS column chromatography.The structures were identified by means of physico-chemical and spectral data.Results From the 70% ethanol extract of the material,four compounds were isolated.Their structures were identified as 11?,12?-di-O-2-methylbutyryl-tenacigenin B (Ⅰ),11?-O-2-methylbutyryl-12?-O-acetyl-tenacigenin B (Ⅱ),tenacissoside H (Ⅲ),and marsdenoside A (Ⅳ),respectively.Conclusion Compound Ⅰ is a new C21 steroid compound,named tenacissoside O.
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Objective:To search for carrier of short active peptide with binding to human CD59 of the surface of HeLa cell.Methods:Ph. D. 12 display peptide library was biopanning with HeLa cell high-expressing CD59. After competitive test, positive phage clones were identified by sandwich ELISA and sequenced.Results:10 phage clones were identified to have higher combination with CD59.Conclusion:The sequence of peptide with binding to CD59 has been obtained.That might lay a basis for design of short peptide of CD59 on tumor sneaking through.
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Objective: To amplify two human mutant CD59 eukaryotic expressing systems and investigate mutant CD59 functional activity. Methods: Mammalian expression vector PLATER of mutant CD59 cDNAs was transfected into CHO together with the pcDNA by lipofectamine,which confered resistance to G418(400 ?g/ml). The positive clones were tested by FIH. Activity of both mutants CD59 was determined by BCECF release assay. Results: Mutant CD59 cDNAs subcloned into the mammalian expression vector PLATER and transfected CHO together with the pcDNA,which confered resistance to G418. The positive clones were tested by FIH.Activity of both mutants CD59 before and after glycation was determined by BCECF release assay,both of them could restrict MAC formation ,and glycation could inhibit CD59. Conclusion: A eukaryotic system that expressing mutant CD59 cDNA was successfully set up.It was found that mutant CD59 could restrict MAC formation,and glycation could inhibit mutant CD59. These would be helpful for the furthur study of link mutant CD59 and the vascular proliferative of diabetes.