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1.
Chinese Journal of Experimental Ophthalmology ; (12): 50-54, 2020.
Article in Chinese | WPRIM | ID: wpr-865223

ABSTRACT

Objective To evaluate the clinical effects of vitrectomy with or without fovea-sparing internal limiting membrane peeling on macular foveoschisis (MF) secondary to pathologic myopia.Methods A non-randomized controlled study was adopted.Twenty-three patients (25 eyes) with refractive error ≥-8.00 DS and MF either combined with foveal retinal detachment or epi-macualr membrane or lamellar macular hole.The subjects were divided into non-internal limiting membrane peeling group (11 patients / 11 eyes) who underwent triamcinolone (TA) assisted vitrectomy and fovea-sparing internal limiting membrane peeling group (12 patients/14 eyes) who underwent TA assisted vitrectomy with fovea-sparing internal limiting membrane peeling.The baseline data such as age,best corrected visual acuity (BCVA),refractive error,axial length were not significant difference between the two group.Main outcomes were BCVA,remission of MF defined by optical coherence tomographyc OCT as well as complications.This study followed the Declaration of Helsinki and this protocol was approved by Ethic Committee of Xiamen Eye Center of Xiamen University (NO.XMYKZX-2016-YWS-007).Results All patients completed follow-up for more 6 months.BCVA (LogMAR) was 0.47±0.30 in non-internal limiting membrane peeling group and 0.40-±0.33 in fovea-sparing internal limiting membrane peeling group,showing no significant difference between the two groups (t =0.66,P=0.52).Complete remission of MF was achieved in 22 eyes.The remission time in non-internal limiting membrane peeling group was 2.5 (1.8,9.3) months,and 1.0 (1.0,3.8) months in fovea-sparing internal limiting membrane peeling group,no statistical significance was obtained between the two groups (U =35.00,P =0.09).One eye had post-operative macular hole in non-internal limiting membrane peeling group,accounting for 9%,while in fovea-sparing internal limiting membrane peeling group,one eye had macular hole,accounting for 7%,and one eye had rhegmatogenous retinal detachment post-operatively,accounting for 7%.Conclusions MF can be resolved by vitrectomy while complete remission can achieved more quickly when combined with fovea-sparing internal limiting membrane peeling.

2.
Chinese Journal of Experimental Ophthalmology ; (12): 50-54, 2020.
Article in Chinese | WPRIM | ID: wpr-798746

ABSTRACT

Objective@#To evaluate the clinical effects of vitrectomy with or without fovea-sparing internal limiting membrane peeling on macular foveoschisis (MF) secondary to pathologic myopia.@*Methods@#A non-randomized controlled study was adopted.Twenty-three patients (25 eyes) with refractive error ≥-8.00 DS and MF either combined with foveal retinal detachment or epi-macualr membrane or lamellar macular hole.The subjects were divided into non-internal limiting membrane peeling group (11 patients /11 eyes) who underwent triamcinolone (TA) assisted vitrectomy and fovea-sparing internal limiting membrane peeling group (12 patients/14 eyes) who underwent TA assisted vitrectomy with fovea-sparing internal limiting membrane peeling.The baseline data such as age, best corrected visual acuity (BCVA), refractive error, axial length were not significant difference between the two group.Main outcomes were BCVA, remission of MF defined by optical coherence tomographyc OCT as well as complications.This study followed the Declaration of Helsinki and this protocol was approved by Ethic Committee of Xiamen Eye Center of Xiamen University (NO.XMYKZX-2016-YWS-007).@*Results@#All patients completed follow-up for more 6 months.BCVA (LogMAR) was 0.47±0.30 in non-internal limiting membrane peeling group and 0.40±0.33 in fovea-sparing internal limiting membrane peeling group, showing no significant difference between the two groups (t=0.66, P=0.52). Complete remission of MF was achieved in 22 eyes.The remission time in non-internal limiting membrane peeling group was 2.5 (1.8, 9.3) months, and 1.0 (1.0, 3.8) months in fovea-sparing internal limiting membrane peeling group, no statistical significance was obtained between the two groups (U=35.00, P=0.09). One eye had post-operative macular hole in non-internal limiting membrane peeling group, accounting for 9%, while in fovea-sparing internal limiting membrane peeling group, one eye had macular hole, accounting for 7%, and one eye had rhegmatogenous retinal detachment post-operatively, accounting for 7%.@*Conclusions@#MF can be resolved by vitrectomy while complete remission can achieved more quickly when combined with fovea-sparing internal limiting membrane peeling.

3.
Chinese Journal of Ocular Fundus Diseases ; (6): 529-533, 2019.
Article in Chinese | WPRIM | ID: wpr-824879

ABSTRACT

Objective To observe the clinical efficacy of digital 3D heads-up display viewing system (3D viewing system) and intraoperative OCT (iOCT) in vitrectomy for myopic foveoschisis (MF).Methods A retrospective,consecutive case series.From October 2018 to May 2019,Nineteen eyes of 19 consecutive patients with MF diagnosed in Xiamen Eye Center of Xiamen University who underwent vitrectomy were included in this study.There were 7 males and 12 females,with the mean age of 54.47± 11.38 years.The average axial length was 30.40±2.30 mm,the mean logMAR BCVA was 0.56±0.31,the mean central foveal thickness (CFT)was 317.80± 151.9.32 μm,the mean max retinal thickness (maxRT) was 556.7 ± 143.7 μm.All the surgeries performed combined with 3D viewing system with iOCT.The standard 25G pars planar vitrectomy were performed with removing the posterior vitreous and indocyanine green (ICG) staining of internal limiting membrane (ILM) and air-fluid exchange.Thirteen of 19 eyes underwent fovea-sparing ILM peeling and the other 6 eyes not.The average follow-up was 4.2 ± 1.4 months.All the patients were on regular follow-up to document the changes on BCVA,anatomical changes in macula,CFT and maxRT.Paired t test was used to compare BCVA,CFT and maxRT before and after surgery.Results The fine images of macula were clearly shown on the 3D viewing system in all eyes.The electronic green filter enhanced the contrast sensitivity of ICG stained images.Clear images of macula were captured by iOCT in all eyes.The average surgical time was 35.5± 8.2 min.On the last follow-up,16 of 19 eyes with MF resolved.The mean CFT was 178.5 ± 103.5 μm,the maxRT was 341.8 ± 83.8.16 μm,and the mean logMAR BCVA was 0.35 ± 0.22.The differences of CFT,maxRT and logMAR BCVA before and after surgery were statistically significant (t=4.181,7.154,5.129;P< 0.001).Minimal invisible full thickness macular hole were detected in 2 eyes by iOCT and repaired with auto serum or ILM flap covering.There was no complication associated with the 3D viewing system.Conclnsions 3D viewing system provides improved contrast and crystal clear macular image stain with ICG in pathological myopia,iOCT can detect the minimal invisible full thickness macular hole during surgery.Both may contribute to improved MF closure rate and BCVA.

4.
Chinese Journal of Ocular Fundus Diseases ; (6): 529-533, 2019.
Article in Chinese | WPRIM | ID: wpr-805488

ABSTRACT

Objective@#To observe the clinical efficacy of digital 3D heads-up display viewing system(3D viewing system) and intraoperative OCT (iOCT) in vitrectomy for myopic foveoschisis (MF).@*Methods@#A retrospective, consecutive case series. From October 2018 to May 2019, Nineteen eyes of 19 consecutive patients with MF diagnosed in Xiamen Eye Center of Xiamen University who underwent vitrectomy were included in this study. There were 7 males and 12 females, with the mean age of 54.47±11.38 years. The average axial length was 30.40±2.30 mm, the mean logMAR BCVA was 0.56±0.31, the mean central foveal thickness (CFT) was 317.80±151.9.32 μm, the mean max retinal thickness (maxRT) was 556.7±143.7 μm. All the surgeries performed combined with 3D viewing system with iOCT. The standard 25G pars planar vitrectomy were performed with removing the posterior vitreous and indocyanine green (ICG) staining of internal limiting membrane (ILM) and air-fluid exchange. Thirteen of 19 eyes underwent fovea-sparing ILM peeling and the other 6 eyes not. The average follow-up was 4.2±1.4 months. All the patients were on regular follow-up to document the changes on BCVA, anatomical changes in macula, CFT and maxRT. Paired t test was used to compare BCVA, CFT and maxRT before and after surgery.@*Results@#The fine images of macula were clearly shown on the 3D viewing system in all eyes. The electronic green filter enhanced the contrast sensitivity of ICG stained images. Clear images of macula were captured by iOCT in all eyes. The average surgical time was 35.5±8.2 min. On the last follow-up, 16 of 19 eyes with MF resolved. The mean CFT was 178.5±103.5 μm, the maxRT was 341.8±83.8.16 μm, and the mean logMAR BCVA was 0.35±0.22. The differences of CFT, maxRT and logMAR BCVA before and after surgery were statistically significant (t=4.181, 7.154, 5.129; P<0.001). Minimal invisible full thickness macular hole were detected in 2 eyes by iOCT and repaired with auto serum or ILM flap covering. There was no complication associated with the 3D viewing system.@*Conclusions@#3D viewing system provides improved contrast and crystal clear macular image stain with ICG in pathological myopia. iOCT can detect the minimal invisible full thickness macular hole during surgery. Both may contribute to improved MF closure rate and BCVA.

5.
Progress in Modern Biomedicine ; (24): 4491-4494, 2017.
Article in Chinese | WPRIM | ID: wpr-614881

ABSTRACT

Objective:To explore the clinical effect ofpaishitang combined with tamsulosin hydrochloride on the patient with up per urinary calculi after extracorporeal shock wave lithotripsy (ESWL).Methods:120 cases with upper urinary calculi in our hospital from January 2015 to September 2016 were selected and divided into two groups according to the random number table,60 cases in each group.ESWL was given to both groups of patients and provided with tamsulosin hydrochloride postoperation,then paishitang were additionally given to the patients in the observation group.The clinical effect and changes of serum creatinine (Scr),neutrophil gelatinase as sociated lipocalin (NGAL),cystatin C (Cys-C) and glomerular filtration rate (GFR) levels before and after treatment were compared between two groups.Results:The total effective rate of observation group was 96.67%,which was 86.67% in the control group,no signifi cant difference was found in the total effective rate between the two groups(P<0.05).The stone discharge rate was 95.00% in the observation group,which was significantly higher than that of the control group (P<0.05);the incidence rate of renal colic was 6.67%,which was significantly lower than that of the control group(P<0.05),the stone discharge time and the duration of hematuria were significantly shorter than those in the control group (P<0.01).There was no significant difference in the recurrence rate between the two groups within one year (P>0.05).The serum NGAL and Cys-C levels of both groups were gradually increased while the GFR levels were gradually decreased on the 1st,2nd day postoperation,but all the index mentioned above gradually recovered on the 3rd day postoperation.The levels of NGAL and Cys-C in the observation group were significantly lower than those in the control group on the 1st,3rd day postoperation while the GFR was significantly higher in the observation group than those of control group on the 1st,3rd day postoperation(P<0.01).No significant difference was found in the Scr at different time points postoperation between two groups(P>0.05).Conclusion:Paishitang combined with tamsulosin hydrochloride had significant clinical effect on thpatient with upper urinary calculus after ESWL and could effectively improve the renal injury induced by ESWL.

6.
Chinese Journal of Tissue Engineering Research ; (53)2007.
Article in Chinese | WPRIM | ID: wpr-594602

ABSTRACT

BACKGROUND:It has great significance to understand individual human leucocyte antigen allele(HLA) haplotype and genotype in organ allotransplantation.OBJECTIVE:To analyse HLA allele frequency,haplotype frequency and characteristices in 36 cases of living-related donor kidney transplantation.DESIGN,TIME AND SETTING:The gene polymorphism analysis was performed in the Tissue Typing Laboratory of Organ Transplantation of Ruikang Hospital of Guangxi Traditional Chinese Medical University from January 2007 to June 2008.PARTICIPANTS:Thirty-six cases of living-related donor kidney transplantation,who were relatives of Guangxi Han population.METHODS:HLA-Ⅰantigen was detected by dry plates of monoclonal antibodies,and HLA-Ⅱ antigen was subjected to genotyping using PCR-SSP technique.And then the allele and haplotype frequencies of HLA-A,B,DRB1 were calculated.MAIN OUTCOME MEASURES:HLA haplotype and genotype of Guangxi Han population.RESULTS:A total of 11 HLA-A,24 HLA-B and 13 HLA-DR alleles were detected among 36 cases,showing a rich polymorphism.A2-B46,A11-B60,B46-DRB1*09,B60-DRB1*15 and A2-B46-DRB1*09 among 144 haplotypes were informative with frequency higher than 0.05.A total of 10 HLA-A-B haplotypes and 9 HLA-B-DRB1 haplotypes were in strongest linkage disequilibrium.Distribution of HLA-A,B,DRB1 alleles and haplotypes in Guanxi population is similar to Han population in South China,such as Hunan and Taiwan,but had its own characteristics.CONCLUSION:HLA alleles have richer polymorphisms with exhibit geographic genetic characteristics.

7.
Chinese Journal of Tissue Engineering Research ; (53): 7701-7705, 2007.
Article in Chinese | WPRIM | ID: wpr-407697

ABSTRACT

BACKGROUND: The quantity and bioactivity of isolated islet are vital to islet transplantation; while, the cold ischemia time and human leucocyte antigen (HLA) typing are key factors to islet quantity and bioactivity which inflect islet transplantation.OBJECTIVE: To observe the effects of cold ischemia time and blood compatibility on quantity and bioactivity of islet cells.DESIGN: Observational study.SETTING: Ruikang Affiliated Hospital of Guangxi College of Traditional Chinese Medicine.MATERIALS: Organs from voluntary donors died of irreversible coma were adopted whose blood-type and HLA typing had been known, pancreases acquisition was carried on after other organs ablation or in the meantime. The blood of identical ABO and HLA matching conformity, or HLA cross-matching hypersensitization (missmatching over 3 Iocuses),or panel reaction antibody (PRA) > 50%, or lymphocyte cytotoxin crossmatching test positive. The isolated and purified islet suspension was filtered by 70 μm filter, which result in preparing 1.2×105/L islet suspention.METHODS: Hypertonic citrate adenine solution was perfused into aorta, and kidney-pancreases and kidney-pancreases-liver were cut together or kidney-pancreases-liver was cut separately. Islet activity was judged by diphenylthiocarbazone (DTZ) dyeing and acridine orange (AO) dyeing; meanwhile, twelve pancreases far from contamination were aquired, mean ablation time was 15 minutes; cold ischemia time ranged from 2.5 to 8 hours. Cold ischemia time of nine pancreases was controlled in 5 hours, warm ischemia time was 0-3 minutes, and peptic time was (15±2.4) minutes, correlation of islet cells and histocompatibility with survival of islet cells was analyzed.MAIN OUTCOME MEASURES: Survival rate of islet cells; counts of platelet, heterophil granulocyte and monocyte.RESULTS: The cutting of kidney, pancreases and liver were successful. If the cold ischemia time was controlled within 5 hours, activity of islets was above 80%. Pancreatic gland used for islet transplantation and cutting of other organs could not affect activity of islet cells. When human islets were exposed to human blood, it would induce a rapid consumption of platelets, neutrophils, monocytes and lymphocytes in the blood. Consumption of blood cells was more in the HLA typing groups than that in the control group. After adding heparin, there was significant difference in cell account among the three groups (P < 0.05) and the reaction was relieved obviously. After 24-hour cultivation, there were significant difference in active islets quantity between HLA typing compatibility group and HLA typing incompatibility group (P < 0.05).CONCLUSION: Pancreatic gland obtained under the cold ischemia time < 5 hours can be used in clinical transplantation of islet cells; a good histocompatibility can raise successful rate of transplantation of islet cells.

8.
Journal of Third Military Medical University ; (24)2003.
Article in Chinese | WPRIM | ID: wpr-561897

ABSTRACT

Objective To investigate the hemostatic effects and safety of the QuGouCao chloroform-water extract.Methods The hemostatic property of the extract was studied by applying the extract soaked gauze to the local wound surface of skin,liver and femoral artery in rabbits.LD50 test in Kuming mice was performed and the tissues of liver and kidney were collected for routine histochemical examination.The skin stimulus in rabbits and the sensitivity test in guinea pigs were conducted.Results The hemostatic time of wounds was obviously shorter(P

9.
Journal of Third Military Medical University ; (24)2003.
Article in Chinese | WPRIM | ID: wpr-678617

ABSTRACT

Objective To investigate the properties of the serum free binding of vascular endothelial cells(ECs) to lipopolysaccharide (LPS). Methods The binding of ECs to fluorescein isothiocyanate labelled LPS(FITC LPS) was observed with laser confocal microscope(LCM). The extract of ECs membrane after binding with FITC LPS was recorded with fluorescence microscope. Results The binding of ECs to FITC LPS was in a dose dependent manner. The fluorescence was mainly distributed in cytoplasm, especially near the nucleus which could also be stained. The bound FITC LPS had a very close relationship with the membranous system of ECs. Conclusion ECs could bind to LPS directly and could function intracellularly.

10.
Journal of Third Military Medical University ; (24)2003.
Article in Chinese | WPRIM | ID: wpr-678578

ABSTRACT

Objective To obtain the protein interacting with inhibitor of differentiation1′(Id1′). Methods The recombinant bait plasmid pHybLex/Zeo Id1′ was constructed and transformed into yeast strain EGY48/ pSH18 34 to test pHybLex/Zeo Id1′ for non specific activation. Adult human lung cDNA libraries were screened to obtain true positive library plasmid. The true positive library clone was obtained by sequencing and basic local alignment sequence tool (BLAST). Results The recombinant bait vector, named as pHybLex/Zeo Id1′, was confirmed by sequencing. pHybLex/Zeo Id1′ was transformed into yeast strain EGY48/pSH18 34 and the transformants had no autonomously activated reporter genes. One true positive clone, obtained by screening of the adult human lung cDNA libraries, was confirmed to be Fyn by sequencing and BLAST. Conclusion Id1′ can interact with Fyn.

11.
Journal of Medical Postgraduates ; (12)2003.
Article in Chinese | WPRIM | ID: wpr-590314

ABSTRACT

Objective: To investigate the expression changes of Bcl-2-associated athanogene 1(BAG-1) and its regulatory effect on the glucocorticoid receptor(GR) activity in rat alveolar macrophages in conditions of cell inflammation and glucocorticoid therapy.Methods: The expression changes of BAG-1 were detected by Western blot after lipopolysaccharide(LPS) and Dexamethasone(Dex) treatment of rat alveolar macrophages(AMs),the interaction between BAG-1 and GR determined by immune coprecipitation experiment,and the transcriptional activation of GR measured by relative luciferase activity assay.Results:After LPS and Dex treatment,the expression of BAG-1L in total protein increased but that of BAG-1S remained changed,BAG-1L rather than BAG-1S was detected in nuclear protein and its expression increased gradually within 24 hours,the interaction between BAG-1L and GR was observed in nucleoli,and the transcriptional activation of GR decreased,with a negative correlation between BAG-1L expression and GR activity.Conclusion:LPS and Dex acting on rat alveolar macrophages,the expression of BAG-1L increases,which,coupled with GR,translocates into nucleoli and inhibits GR activity.This might be the important mechanism that underlies glucocorticoid resistance in inflammation.

12.
Chinese Journal of Burns ; (6): 170-172, 2002.
Article in Chinese | WPRIM | ID: wpr-289217

ABSTRACT

<p><b>OBJECTIVE</b>To screen the optimal transfection method for human umbilical vein endothelium, so as for us to establish a basic method for the further transfection with other objective genes.</p><p><b>METHODS</b>The pEGFP-Actin plasmid was employed as an ectogenetic gene and was transfected into ECV-304 endothelium cells by lipofection, DEAE-dextran or electroporation transfection methods, respectively. The cell death rate and transfection rate after the transfection were compared among the three methods.</p><p><b>RESULTS</b>(1) Twelve hours after the transfection, the cell death rate was 4% by all three methods, however, the cell death rate increased up to 50% at 60 hours after the transfection by DEAE-dextran. (2) The transfection rate of ECV-304 could reach 95% by all the three methods. But the fluorescent intensity was stronger in lipofection group compared with electroporation method.</p><p><b>CONCLUSION</b>Gene transfection into ECV-304 with lipofection and electroporation methods exhibited better stability and repeatability.</p>


Subject(s)
Humans , Actins , Genetics , Cells, Cultured , Electroporation , Endothelium, Vascular , Physiology , Genetic Vectors , Genetics , Plasmids , Genetics , Recombinant Fusion Proteins , Genetics , Transfection , Methods
13.
Chinese Journal of Burns ; (6): 279-281, 2002.
Article in Chinese | WPRIM | ID: wpr-289196

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the significance of the expression pattern and its signal modulating mechanism of endothelial ICAM-1 induced by LPS.</p><p><b>METHODS</b>(1) The expression pattern of the ICAM-1 was observed at mRNA level in cultured human umbilical vascular endothelial cells (HUVECs) strain ECV-304 after being stimulated by different LPS concentrations at different time points. (2) The modulating effects of different signal pathways on the ICAM-1 expression of the HUVECs were observed at mRNA and protein levels under the stimulation of LPS after the cells were primed by signal pathway blocking agents for 30 mins.</p><p><b>RESULTS</b>(1) The ICAM-1 mRNA expression could be induced by LPS (100 pg/ml) for 6 hours, and the expression was enhanced along with the increase of LPS concentration. The expression peaked when LPS was at concentrations of 100 approximately 1 000 ng/ml. Temporally, the mRNA expression reached the top level at 6 approximately 8 hours and remained high 12 hours after the stimulation. (2) The expressions of ICAM-1 mRNA and protein could be significantly inhibited by PSI, the NF-kappaB inhibitor. Moreover, the expression of ICAM-1 could all be partially inhibited at mRNA and protein levels by PD98059, the ERK1/2MAPK inhibitor, as well as SB203580, the p38MAPK inhibitor.</p><p><b>CONCLUSION</b>The ICAM-1 mRNA expression of HUVECs could be induced by LPS in both dose and time dependent manner. NF-kappaB might be the major signal pathway of modulating ICAM-1 expression, and p38 and ERK1/2 could possibly be signal pathways of minor importance.</p>


Subject(s)
Humans , Cells, Cultured , Dose-Response Relationship, Drug , Gene Expression Regulation , Intercellular Adhesion Molecule-1 , Genetics , Lipopolysaccharides , Toxicity , Mitogen-Activated Protein Kinases , Physiology , NF-kappa B , Physiology , Signal Transduction , Time Factors
14.
Medical Journal of Chinese People's Liberation Army ; (12)2001.
Article in Chinese | WPRIM | ID: wpr-554096

ABSTRACT

In order to explore the effects of exogenous human telomerase reverse transcriptase (hTERT/hTRT/hEST2) on telomeric restriction fragment (TRF), telomerase activity and its subunits expression in human embryonic fibroblasts (hEFs), hTERT sense eukaryotic expression vector pIRES2 EGFP hTERT was constructed with DNA recombinant technique and then transfected into primary hEFs by Lipofectin method. TRF length, telomerase activity and changes in telomerase subunits expression were examined and evaluated in transfected and untransfected cells. The results showed that telomerase activity in pIRES2 EGFP hTERT transfected cells (hEF EGFP) was significantly higher than that in untransfected hEFs and vacant vector transfected cells (hEF EGFP) ( P

15.
Journal of Chinese Physician ; (12)2001.
Article in Chinese | WPRIM | ID: wpr-521405

ABSTRACT

Objective The recombinant fluorescent eukaryotic expressing vector containing hTERT cDNA was transfected into human embryonic fibroblasts (hEFs) to explore the effects of exogenous hTERT on the type I and III collagens expression in hEFs. Methods p IRES2-EGFP-hTERT plasmid and pIRES2-EGFP plasmids were transfected into primary hEFs respectively by Lipofectin reagent. Expression of type I and III collagen was determined by Western blotting and the content of type I and III collagens in the cellular medium at 3 days after transfection were examined by radio-immunoassay. Results The expression levels of type I and III collagens in hTERT gene transfected hEFs(hEF-hTERT) were obviously higher than those in untransfected hEFs and vacant vector transfected hEFs(hEF-EGFP). The content of type I and III collagens in the cellular medium in hEF-hTERT cells at 3 days after transfection was also higher than that in untransfected hEFs and hEF-EGFP cells. Conclusions The synthesis ability of type I and III collagens in hEFs could be promoted by exogenous hTERT gene transfection.

16.
Journal of Third Military Medical University ; (24)1983.
Article in Chinese | WPRIM | ID: wpr-678121

ABSTRACT

Objective To screen and analyze genes up regulated in human umbilical vein endothelial cells (HUVEC) stimulated by lipopolysaccharide (LPS). Methods Suppression subtractive hybridization (SSH) was performed between the unstimulated HUVEC(driver) and HUVEC stimulated with LPS(tester) to generate subtractive cDNA library. The library was screened with colony dot hybridization to further verify the differentially expressed cDNA clones. Positive clones were sequenced and BLAST analyzed. The 3 novel cDNA sequences were verified by RT PCR. Results Twenty five up regulated genes related to inflammation, cellular cytoskeletal rearrangement, cellular proliferation and apoptosis, intercellular message transduction, and 3 new expression sequence tags (EST) were acquired. RT PCR indicated the expression of the new ESTs only in HUVEC stimulated by LPS. Conclusion SSH is a powerful technique of high sensitivity for the detection and clone of up regulated gene expressed in HUVEC stimulated by LPS, which may be helpful to clarify the mechanism of endothelial cells activation stimulated by LPS.

17.
Medical Journal of Chinese People's Liberation Army ; (12)1981.
Article in Chinese | WPRIM | ID: wpr-554862

ABSTRACT

Objective To study the subcellular localization and the tissue expression of EOLA1(endothelial-overexpressed lipopolysaccharide-associated factor 1). Methods The fusion protein EOLA1-EGFP expressed vector was constructed and transfected into endothelial cells. After 24 hours posttransfection, the subcellular localization of EOLA1 was detected by laser-scanning microscopy. The tissue-specific distribution of EOLA1 was assessed with Multiple Tissue Northern Blots. Results The expression of EOLA1 was tissue-specific in various human tissues. With human multiple tissue Northern blot analysis, it was shown that EOLA1 could express in the heart, skeletal muscle, kidney, liver, placenta, colon, spleen, small intestine, but did not in the brain, lung, thymus, and peripheral blood leukocyte. EOLA1 was mainly localized in cytoplasm and could move into the nucleus. Conclusion EOLA1 is one of intercellular proteins and may play a role in intercellular signal transduction.

18.
Medical Journal of Chinese People's Liberation Army ; (12)1981.
Article in Chinese | WPRIM | ID: wpr-552726

ABSTRACT

To screen genes in endothelial cells resulted from responding to lipopolysaccharide (LPS), mRNA was extracted from both untreated human umbilical endothelial cells (HUVEC) following and HUVEC which were treated with LPS for 6 hours. cDNAs of both populations were synthesized to generate cDNA libraries by suppression subtractive hybridization (SSH). The libraries were then screened with colony dot blots. Positive clones were sequenced and BLAST analysed. The results showed differential genes included 3 novel genes and 22 known genes. The 3 novel genes were confirmed by Northern blotting analysis. These 22 known genes were involved in the regulation of proinflammatory response, cell apoptosis, cytoskeleton, signal transduction and energy metabolism. These results suggest that SSH is an effective technique to detect differential gene expression in HUVEC, which may be helpful to evaluate molecular mechanisms of endothelial injury induced by LPS and provide potential therapeutic targets for LPS related disturbances.

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