Aim Vascular endothelial cell growth inhibitor(VEGI) is a recently discovered novel member of the TNF superfamily,which is expressed predominantly in endothelial cells.As an endothelial cell-specific negative regulator of angiogenesis,the relationship between structure and function of VEGI is not understood at present.Methods In order to explore the functional key amino acids of VEGI,four mutants of VEGI(E45→R,G47→A,Y111→F,Y111→T) were construced by site-directed mutagenesis,and recombinant proteins were generated from E.coli.Four mutant proteins behaved similar to the wild type VEGI in various physico-chemical assays.The proliferation of HUVEC and chick choriallantic membrane assay were performed to study the activity of four mutants.Results The mutant E45→R significantly decreased the biological activity,and the mutant G47→A caused a slight drop on activity,but the mutants Y111→F,Y111→T almost completely abolished biological activity.Conclusion It suggests that Y111 is an important residue in biological activity,which may play a direct role in receptor recognition.Moreover,the tyrosine ring and hydroxy group of the amino acid are important determinant of biological activity.Additionally,E45 also plays an important role in biological activity of VEGI.

Objective To investigate the effects of social support intervention on anxiety and depression of pulmonary hypertension(PH)patients,then provide a scientific basis for nursing of patients with pulmonary hypertension.Methods The general condition of PH questionnaire,self-rating anxiety scale (SAS),self-rating depression scale(SDS),social support rating scale(SSRS)were distributed to 131 patients with PH.Then make statistical analysis of patients'anxiety,depression and social support conditions.Results The score of anxiety and depression psychological conditions of patients with pulmonary hypertension was significantly higher than normal population,the difference wag statistically significant.Among 131 patients,16 patients with anxiety,accounting for 12.21% ;21 cases of patients with depression,accounting for 16.03% .28 patients at a high level of social support,92 patients at a medium level of social support,11 patients at a low level of social support,a total of 91.60% of the patients in the middle and higher levels of social support.Anxiety and depression scores had significant negative correlation with social support,objective support points and subjective support points.The anxiety,depression difference among different types of pulmonary hypertension was statistically significant.The difference of anxiety and depression scores between patients with idiopathic pulmonary arterial hypertension and patients with pulmonary hyper tension caused by congenital heart disease were significant.The depression scores between pulmonary hy pertension caused by pulmonary veno-occlusive disease and congenital heart disease were significantly different.Conclusions When nurses care pulmonary hypertension patients.those with different types of PH should be given targeted social support.Attention should be paid to transfer the source of social support to help them adopt a positive attitude to face the diseabe,then improve the treatment and care compliance of patients.

Objective To investigate the effects of sevoflurane postconditioning on myocardial ischemiareperfusion(I/R)injury in patients undergoing coronary artery bypass grafting(CABG)with cardiopulmonary bypass(CPB).Methods Forty NYHA Ⅰ -Ⅲ patients of both sexes,aged 55-64 yr,with BMI ＜ 30 kg/m2,scheduled for CABG under CPB,were randomly divided into 2 groups(n = 20): control group(group C)and sevoflurane postconditioning group(group S).Anesthesia was induced with midazolam and/or etomidate,fentanyl and rocuronium.Patients were tracheal intubated and mechanically ventilated.Anesthesia was maintained with iv infusion of propefol and intermittent iv injection of fentanyl and pipecuronium.In group S,2% sevoflurane was inhaled continuously for 15 min immediately after aortic unclamping.After anesthesia induction,before CPB,10 min after the end of CPB,at the end of operation,and 6 and 24 h after operation,MAP,HR,CVP,mean pulmonary arterial pressure,pulmonary arterial wedge pressure,CO and S(v)O2 were recorded,and CI,SVI,systemic vascular resistance index and pulmonary vascular resistance index were calculated.Blood samples were taken from central vein before aortic clamping,at 6 h of reperfusion and 24 h after operation for determination of plasma creatine kinase(CK),creatine kinase isoenzyme(CK- M B)and lactate dehydrogenase(LDH)activities and tropenin I(TnI)concentrations.Myocardial tissues were obtained from right auricle before aortic clamping and at the end of CPB for observation of the ultrastructure and the severity of myocardial injury was assessed.Results There was no significant difference in hemodynamics and parameters of cardiac function between the two groups(P ＞ 0.05).Compared with group C,plasma CK-MB and LDH activities at 6 h of reperfusion and plasma CK activity and TnI concentrations at 24 h after operation were significantly decreased and the myocardial injury was significantly reduced after the end of CPB in group S(P ＜ 0.05).Conclusion Sevoflurane postconditioning can protect myocardium against I/R injury induced by CPB in patients undergoing CABG.

Objective To assess the application value of disposable syringe of low resistance (SLR) in epidural puncture.Methods One hundred and thirty-two patients scheduled to undergoing selective operation under epidural or combined epidural-spinal anesthesia were divided into glass syringe (GS) group and SLR group with 66 cases each by random digits table method.GS was applied in local anesthesia and epidural puncture with loss of resistance in GS group.While SLR was used in SLR group.The defect of the syringes' appearance was evaluated.The fastness between each sub-unit during the procedure was observed.The rupture,dropping,leakage,air leakage and blockage were also estimated.The heart rate,blood pressure,oxygen saturation and other adverse events were also observed.Results In SLR group,syringe excepted 1 case of slight resistance but did not affect the use,the rest of the components connected firmly,surface without defects,clear scale uniform color,not observed rupture,air leakage,leakage and blockage phenomenon,pump liquid and air resistance was small.In GS group,syringe surface without defects,clear scale uniform color,3 cases had loose connection,1 case with rupture when using,5 cases with air leakage and leakage,3 cases of pumping liquid or dry air resistance to 0.9％ sodium chloride and moist after use.There was significant difference in appearance and usage between two groups (x2 =10.324 5,P =0.001 3).There was no significant difference in a puncture success,first three puncture success and inject air negative pressure sensitivity between two groups (P ＞ 0.05).Conclusion SLR is worth to be applied in epidural puncture.

OBJECTIVETo investigate the role of CD40 ligand (CD40L) in dendritic cells (DC) of CEA transgenic mice and to evaluate the specific cellular immunity induced by activated DC.

METHODSBone marrow cells of the CEA transgenic mice were used to generate immature dendritic cells under the condition of GM-CSF and IL-4. CD40L was added to activate dendritic cells into mature phenotype. Dendritic cells cancer vaccine was pulsed with CEA526-533 peptide which made the vaccine specific for cancer immunity. The immunophenotype molecules were identified by flow cytometry. The cytokines produced by cells were determined by ELISA. T cells proliferation was measured by (3)H-thymidine essays.

RESULTSImmunophenotype molecules expressions of CD40L-activated dendritic cells were significantly higher than those in control group. IL-12 secretion by CD40L-activated dendritic cells was (937.81+/-51.99) pg/10(6) DC, significantly higher than that in control group [(83.06+/-8.58) pg/10(6) DC, P<0.01]. CD8(+) T cells proliferation induced by CD40 L-activated dendritic cells was stronger as compared to control group (P<0.05), and the secretion of IFN-gamma was(33.900+/-4.550) ng/L, significantly higher than that in control group [(5.226+/-0.460) ng/L, P<0.01]. Splenocytes proliferation induced by CD40 L-activated dendritic cells was stronger as compared to control group (P<0.01), and the secretion of IFN-gamma was (69.802+/-11.407) ng/L, significantly higher than that in control group [(2.912+/-0.562) ng/L, P<0.01].

CONCLUSIONThe method of using CD40L to stimulate bone marrow-delivered dendritic cells promotes the maturation and activation of dendritic cells, which enhances the cellular immunity in CEA transgenic mice.

Animals ; CD40 Ligand ; immunology ; physiology ; Dendritic Cells ; cytology ; immunology ; metabolism ; Immunity, Cellular ; immunology ; Mice ; Mice, Inbred C57BL ; Mice, Transgenic

OBJECTIVETo investigate the dynamic expressions of TGF-beta 1 and TGF-beta 1 mRNA at different stages of hepatocellular carcinoma (HCC) development and their use in clinical diagnosis.

METHODSHepatoma models were developed with 2-FAA using male Sprague-Dawley (SD) rats. Morphological changes of the rat liver histological preparations (H and E stained) were studied. The fragment of TGF-beta 1 gene obtained was amplified by nested RT-PCR. Dynamic change of TGF-beta 1 level was quantitatively analyzed by ELISA. The distribution of TGF-beta 1 in the cells and its gene expression were detected in human HCC tissues.

RESULTSThe progressive increases of hepatic TGF-beta 1 and TGF-beta 1 mRNA were observed in rat hepatocytes which progressed from granular degeneration, atypical hyperplasia and finally to HCC development induced by 2-FAA. The expression levels in HCC tissues were significantly higher than those in the normal and degenerative ones. TGF-beta 1 was shown in rat hepatocytes by immunohistochemistry. Plasma TGF-beta 1 was detected in 89.5% of all the patients with HCC, but it was detected in 93.3% of them who had an AFP less than 400 microg/L. TGF-beta 1 mRNA showed a stronger expression in HCC tissues. TGF-beta 1 mRNA was found in peripheral blood mononuclear cells from all HCC patients with extrahepatic metastasis.

CONCLUSIONTGF-beta 1 may participate in hepatocyte canceration. The overexpression of TGF-beta 1 and TGF-beta 1 mRNA could be useful markers for early diagnosis and predicting prognosis of HCC.

Adult ; Aged ; Animals ; Biomarkers, Tumor ; blood ; Carcinoma, Hepatocellular ; blood ; diagnosis ; pathology ; Female ; Humans ; Liver Neoplasms, Experimental ; blood ; diagnosis ; pathology ; Male ; Middle Aged ; Neoplasm Metastasis ; prevention & control ; Prognosis ; RNA, Messenger ; blood ; Rats ; Rats, Sprague-Dawley ; Transforming Growth Factor beta1 ; blood

OBJECTIVETo investigate the effects of Annexin A2 (ANXA2) deficiency on the malignant biological behaviour of hepatoma cells.

METHODSThe human hepatocellular carcinoma (HCC) cells lines MHCC97-H, HepG2, SMMC-7721, SMMC-7402 and L02 were evaluated. The expression and distribution of ANXA2 were analysed by western blotting, real-time PCR, immunofluorescence and immunohistochemistry.Cell cycle was assessed by flow cytometry and propidium iodide staining. Effects of ANXA2 silencing on invasion and migration potential were assessed by transwell assay and wound healing assay, respectively. Proliferative potential was assessed by CCK-8 kit in vitro and xenograft tumour-growth assay in vivo. The t-test, chi square test, rank sum test, q-test and F-test were used for statistical analyses.

RESULTSThe expression level of ANXA2 was markedly higher in the MHCC97-H cells with high metastasis potential than in the HepG2, SMMC-7721, SMMC-7402 and L02 cells. The efficiency of shRNA-mediated ANXA2 deficiency was more than 80%. Immunofluorescence analysis of the MHCC97-H cells indicated that ANXA2 expression was mainly localized to the cellular membrane and cytoplasm, with some nuclear localization. Down-regulation of ANXA2 led to S-phase arrest of HCC cells (q =8.001, P =0.002) and an inhibition of proliferation (q =17.140, P less than 0.01), migration (q =12.808, P less than 0.01) and invasion potential (q =9.069, P =0.002). Xenograft tumour-growth assay indicated that shRNA targeting of ANXA2 led to lower tumour weight (q =11.968, P < 0.001) and down-regulated ANXA2 expression (Z =2.530, P =0.011).

CONCLUSIONDown-regulation of Annexin A2 gene transcription effectively changes the biological behaviours of hepatoma cells, and may represent a potential target of HCC molecular therapies.

Animals ; Annexin A2 ; genetics ; Carcinoma, Hepatocellular ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Humans ; Liver Neoplasms ; pathology ; Neoplasm Transplantation ; RNA, Small Interfering ; genetics ; Signal Transduction ; Transcription, Genetic

Background Macular hole retinal detachment(MHRD) causes severe decrease of vision.Pars plana vitrectomy has been applied for MHRD.Silicone oil and C3F8 have been used as intraocular tamponade materials,while there are still controversity on their efficacy.Objective This trial was to evaluate the efficacy of vitreous surgery combined with silicone oil or C3F8 tamponade for patients with MHRD in highly myopic eyes.Methods A retrospective case-controlled study was designed.Fifty-one patients who underwent vitrectomy with intraocular tamponade were retrospectively analyzed.The best corrected visual acuity(BCVA) before and after surgery were examined and campared between two groups.The rate of recurrent RD and macular hole closed were analyzed.Results There was no significant difference in BCVA between silicone oil group and C3F8 tamponade group before and after surgery(U=266.000,P =0.286 ; U =205.000,P =0.029).The BCVA after surgery was elevated in both groups (Z=-2.729,P =0.006 ; Z =-3.273,P =0.001).The rates of recurrent RD and macular hole closed were no difference between two groups(P=0.894,1.000).There were no other complications but cataract.Conclusions C3F8 tamponade of MHRD in high myopic eyes show the same efficacy as silicone oil tamponade.

Adenocarcinoma ; genetics ; Adult ; Age Factors ; Aged ; Base Sequence ; Carcinoma, Squamous Cell ; genetics ; DNA, Mitochondrial ; genetics ; Female ; Humans ; Lung Neoplasms ; genetics ; Male ; Middle Aged ; Sequence Deletion ; Smoking ; genetics

OBJECTIVEGuanxin II is a famous modern formula of traditional Chinese medicine. Guanxin II after myocardial infarction (MI) has been shown to have beneficial effects on cardiac anatomy and function. The purpose of this study was to examine the effects of Guanxin 1I on cardiac protein expression after MI.

METHODRats were randomized into 3 groups, sham, model and treating groups. Model and treating groups were fed with Guanxin II and sham group was fed with water for 10 days before MI. MI operation is to ligate left coronary artery. 24 hours after MI, myocardial protein expression of junctional zone was assessed with 2D gel electrophoresis and mass spectra analysis.

RESULTGuanxin II was found to be able to improve myocardial protein expression, especially 11 proteins. These proteins are mainly involved in suppressing changes of cell shape and structure and energy metabolism.

CONCLUSIONGuanxin II after MI affected myocardial protein expression. Further experiments of larger research extent should be done to receive more results.

Animals ; Cell Shape ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Energy Metabolism ; drug effects ; Gene Expression Regulation ; drug effects ; Heart ; drug effects ; Male ; Muscle Proteins ; metabolism ; Myocardial Ischemia ; metabolism ; pathology ; Myocardium ; metabolism ; Rats