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Background and purpose: Gene chip is a nucleic acid sequence analysis method which is based on hybridization. It is a high-through put assay which can widely detect the level of gene expression in different tissues and cell types. This study aimed to compare and bioinformatically analyze differentially expressed genes between higher malignant degree of prostate cancer tissues and prostate inflammation tissues. Methods: The total RNAs were isolated from tissues of prostate cancer and prostate inflammation by TRIzol method and then purified, reversely tran-scribed to cDNA with incorporating biotin labeling probe, hybridized with Affymetrix Human U133 Plus 2.0 (covering 47000 transcripts,representing 38500 distinct genes). Picture signals of fluorescence in gene array were scanned and differential expression of gene in two tissues were compared by Command Console Software 4.0. These differential expressed genes were analyzed by bioinformatics methods finally. Results: According to the fold change ≥2, P<0.05, 1819 differential expression genes including 1025 up-regulated genes and 794 down-regulated genes were discovered. GO enrichment analysis displayed that these differentially expressed genes were mainly involved in cell cycle, cell metabolism, etc. KEGG pathway analysis found that these genes were mainly involved in some metabolism pathways including purine nucleotide metabolism. The interactions between the proteins encoded by these genes were analyzed by STING. Twenty key nodes genes including TPX2, ANLN, NUSAP1, MELK, DLGAP5, KIF11, TOP2A, RRM2 were dis-covered. Then this study revealed CEP55 and ANLN might be related to the occurrence and metastasis of prostate cancer by looking through literature. Conclusion: During the development of prostate cancer, the activation of genes related to cell cycle and cell migration, the abnormalities of genes related to metabolism and the inhibition of genes related to cell adhesion play critical roles in the development of prostate cancer. CEP55 and ANLN were related to the occurrence and prognosis of prostate cancer by systematic analysis which provided a valuable clue for the next experiment.
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Background and purpose:Hypermethylated in cancer 1 (HIC1) is silenced in multiple cancer cells and tissues by DNA methylation of epigenetic modification, which may modulate the initiation and progression of tumors. However, there are few reports about this phenomenon in prostate cancer. This study aimed to investigate the status of HIC1 promoter methylation in prostate cancer using methylation methods.Methods:Methylation-specific polymerase chain reaction (MSP) and bisulfate sequencing PCR (BSP) were used to detect the methylation status ofHIC1 promoter in prostate cancer cell lines PC3 and C4-2B, prostate normal cell line PrEC, primary Chinese PCa tissues and the respective healthy control cases.HIC1 expression level was respectively determined by reverse transcription-PCR (RT-PCR) and Western blot assays in PC3, C4-2B and PrEC cells treated with 5-Aza-CdR.Results:We found that the percentages of HIC1 promoter methylation were 78.23%, 72.15% and 10.63% in PC3, C4-2B and PrEC cells by MSP analyses. Moreover, the levels of methylatedHIC1 promoter in 36 primary Chinese PCa tissues compared with the respective healthy control cases were 80.30%vs 31.56%. Expressions ofHIC1 mRNA and protein level were restored in PC3 and C4-2B cells after 5-Aza-CdR treatment.Conclusion:These findings demonstrate thatHIC1 promoter region is hypermethylated in prostate cancer, which results in silence or downregulation ofHIC1. The status ofHIC1 methylation can be a valuable marker in the early stage of prostate cancer and a potential therapeutic target.
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Objective To study the molecular epidemiology of C.albicans isolates in infectious disease patients and to explore biofilm phenotypic characterization responsible for biofilm formation in clinical strains.Methods A total of 104 hospital-acquired C.alibcans clinical isolates collected from sterile sites and mucosal lesions of 92 infectious disease patients ( viral hepatitis, tuberculosis and AIDS) in Shanghai Public Health Clinical Center were analyzed.MLST analysis was performed to identify their phylogenetic status.The capability of biofilm formation was measured by [2,3-bis-(2-methoxy-4-nitro-5-sulphenyl)-2H-tetrazolium-5-carboxanilide] XTT assay.The results were compared using Kruskal-Wallis test.Results MLST analysis identified 63 DSTs with a decentralized phylogeny among 104 C.albicans isolates, of which 41 DSTs (65.1%) had not been reported in the online MLST database.The Single Locus Sequence Query from the C.albicans database identified new alleles.MEGA6 analysis of the MLST data assigned the 104 isolates within 14 of the 18 known clades; among them the clade 1 contained the greatest proportion of isolates (26.9%).Of the 43 novel DSTs isolates, 37 ( 86.0%) clustered within 11 of the 18 known clades.16 high biofilm formers were found from a total of 104 clinical isolates.The biofilm formation capabilities differed in strains isolated from different anatomical sites (H =18.23,P=0.0326).Biofilm formation by blood-originated isolates was lower than that of catheter-originated isolates ( Z=-72.20,P<0.001).Genotypes also affected the biofilm formation capability of the C.albicans isolates (H=10.01,P=0.0185).Conclusions A high level of diversity within C.albicans isolates.Microevlution clearly influences C.albicans genetic alterations upon environmental selection.The site of isolation and genotype associates with the biofilm formation capability.
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Objective: To investigate the short term clinical efifcacy of endovascular repair for complicated acute type Stanford B aortic dissection. Methods: To retrospectively analyze the clinical data of 36 patients with complicated acute type Stanford B aortic dissection who received endovascular repair in our hospital from 2010-01 to 2014-06 including operational procedure and post-operative follow-up of CT angiography. There were 27 male and 9 female patients with the average age of 43.7 years (41-62) years. Results: Successful operations were conducted in all 36 patients. 22 patients received endovascular repair combined with covering left subclavian artery (LSA),10 received endovascular repair combined with chimney technique, 2 received endovascular repair combined with vascular prosthesis bypass from left common carotid artery to LSA, 2 received endovascular repair combined with vascular prosthesis bypass from right common carotid artery to left common carotid artery, whose proximal parts were ligated. Viscera artery and lower extremity artery supply were restored gradually. No complication of endoleak occurred. There 30/36 (83.33%) patients were followed-up for 1 year, and 10 patients developed thrombus in full false lumen and 20 developed thrombus in partial false lumen after 1 year. Compared with pre-operative values, thoracic aortic true lumen volume increased in either thrombus in full false lumen (190 ± 68.7) ml vs, (125.3 ± 63.4) ml and thrombus in partial false lumen (166.2 ± 71.8) ml vs (110.1 ± 62.7) ml,P Conclusion: For endovascular repair of complicated aortic dissection, covering LSA with chimney technique and hybrid operation of small incision could extend anchor zone and expand the range of endovascular repair which may improve the effect and reduce the complication for good short term effect.
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Objective To investigate strain distribution and antifungal susceptibility of fungus isolates from acquired immune deficiency syndrome (AIDS ) patients of Shanghai public health clinic center .Methods The funus isolates from clinical specimens of in‐hospital AIDS patients in our hospital between January 2010 and December 2014 were retrospectively analyzed . Results Of the 3 155 hospitalized patients with human immunodeficiency virus (HIV)/AIDS ,a total of 11 291 fungus culture specimens were collected ,of which 1 786 (15 .82% ) were positive .Nine hundred and seventy‐nine fungus strains were isolated ,which were identified as 27 species or genus .The most common isolates were Candidaalbicans (503),Candida tropicalis (60),Candida glabrata (48),Candida krusei (41), Cryptococcus neoformans (179) and Penicillium marnef fei (59) .The majority positive samples were from respiratory tract (61 .29% ) ,followed by the feces (13 .28% ) ,cerebrospinal fluid (11 .24% ) and blood (11 .13% ) .The positive isolation rate of sterile tissue specimens (mainly blood and cerebrospinal fluid) was 6 .92% (558/8 052 ) , and 96 .24% (537/558 ) of the isolated fungi were Cryptococcus neoformans and Penicillium marnef fei .The drug susceptibility rate of Candida to five antifungal drugs commonly used in clinical (amphotericin B ,5‐fluorine cytosine ,fluconazole ,itraconazole ,voriconazole) were 100 .00% ,91 .67% ,83 .33% ,70 .83% and 83 .33% ,respectively .The drug susceptibility rate of Cryptococcus neoformans to three antifungal drugs commonly used in clinical (amphotericin B ,5‐fluorine cytosine ,fluconazole) were 96 .05% ,94 .74% and 97 .37% ,respectively .Conclusions The predominant species of fungal pathogens in AIDS patient in our hospital include Candida ,Cryptococcus neoformans and Penicillium marnef fei .The pathogen distribution of blood and cerebrospinal fluid are different .Some of Candida and Cryptococcus neoformans are resistant to the commonly used antifungal drugs .
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Objective To analyze the impact and its mechanism of down-regulation of AnnexinA2 expression on prostate cancer(PCa) invasion and metastasis.Methods The expression of AnnexinA2 in three prostate cancer cell lines with different metastasis ability,including LNCaP (lower metastasis ability),PC3 (lower metastasis ability),C4-2B (higher metastasis ability) were detected by Western blot.The correlations between the expression of AnnexinA2 and the metastasis ability of prostate cancer cells were also evaluated.The siRNA was used in PC3 cells to down-regulate the expression of AnnexinA2,the cell proliferation assay was performed by MTT method,the cell apoptosis level was detected by flow cytometry,the expression of MMP-2 and MMP-9 were detected by Westrn blot,the in vitro invasiveness of PC3 was detected by transwell cabinet test,and the migration ability of PC3 cells was detected by scratch test,respectively.Results The grayscale value of AnnexinA2 expression in C4-2B cells is 0.22,in contrast with the internal reference,which was obviously lower than those of LNCaP and PC3 cells with lower metastasis potency(relative grey value is 0.93 and 0.95,respectively.P＜0.01).After RNAi was used in PC3 cells to down-regulate the expression of AnnexinA2,the growth became faster for PC3-ANXA2-siRNA cells than PC3,PC3-Lip and PC3-empty vector cells (P＜0.05).After RNAi was used in PC3 cells to down-regulate the expression of AnnexinA2,the ratio of apoptosis was detected by flowcytometry in PC3,PC3-Lip,PC3-empty vector and PC3-ANXA2-siRNA cells,and the apoptosis ratio in PC3-ANXA2-siRNA cells was the highest.However,the difference was not significant compare to others (P＞0.05).After RNAi was used in PC3 cells to downregulate the expression of AnnexinA2,the expression of AnnexinA2 in PC3-ANXA2-siRNA cells was decreased while the expression of MMP-2 and MMP-9 were increased.After RNAi was used in PC3 cells to down-regulate the expression of AnnexinA2,invasiveness of PC3-ANXA2-siRNA cells detected by transwell cabinet test was increased in vitro,and migration of PC3-ANXA2-siRNA cells detected by scratch test was increased in vitro.Conclusions The down-regulation of expression of AnnexinA2 could increase the invasive and metastatic ability of prostate cancer,and this may attribute to the up-regulation of MMP-2 and MMP-9 expression in prostate cancer.
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Objective To evaluate the clinical efficacy of oral Sarpogrelate hydrochloride in the treatment of chronic arterial ischemia of the lower extremities.Methods In this study 892 patients,who suffered from arteriosclerosis (ASO) or thromboangiitis obliterans ( TAO ) or diabetic foot ( DF ),with symptoms of intermittent claudication, sensation of cold, pain, ulcer, and without a history of vasotransplantation or bypass grafting or interventional therapy, were treated by taking Sarpogrelate hydrochloride tablets 100 mg tid for consecutive 8 weeks.The improvement rate of concomitant symptoms and the total effective rate of ASO, TAO, DF were evaluated.Drug adverse reaction were recorded.Results The improvement rate of intermittent claudication,sensation of cold,pain and ulcer were 96.9％,97.1％,89.0％ and 86.9％ respectively.The total effective rate for ASO,TAO,DF was 83.5％.A total of 81 cases (9.1％) reported mild side effects,including 7 patients with mild rash after 2- 5 days' medication,21 patients with mild nausea and 53 patients with stomach discomfort after 1 - 2 days' medication.Symptoms were managed conservatively without discontinuing taking sarpogrelate hydrochloride.Conclusions Sarpogrelate hydrochloride oral is a safe and effective therapy for chronic arterial ischemia diseases of the lower extremities.
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Objective To express and purify Chlamydial protease-like activity factor(CPAF)from Chlamydophila pneumoniae,for investigating the effect of its recombinant protein GST-CPAF in inducing human monocytic cells to secrete proinflammatory cytokines and cell apoptosis.Methods The recom-bination expression plasmid pGEX6p-2/CPAF from Chlamydophila pneumoniae was transformed into E.coli.The recombination GST-CPAF was expressed after induction by IPTG,and purified by a agarose gel FF.Human monocytic cells were stimulated by the GST-CPAF to test the production of tumor necrosis factor a(TNF-α)and interleukin-6(IL- 6)by ELISA.Inhibition of cells proliferation with GST-CPAF was assessed by MTT.The THP-1 cell apoptosis stimulated by GST-CPAF was detected by Hoechst33258 fluorescence staining,DNA fragmentation analysis and cell apeptosis was detested bv Annexin V-FITC-propidiuum iodide (PI)staining.Results The recombination protein GST-CPAF was successfully expressed with high level in E.coli,and stimulated human monocytic cells to produce proinflammatory cytokines including TNF-α and IL-6 in a dose-and time-dependent manner.Otherwise,the GST-CPAF inhibited the growth of human monocytic cell in a dose-dependent manner.Apoptosis with nuclear chromatin fragmentation as well as cell shrinkage was observed by fluorescent staining and microscopy,DNA ladders in apoptosis cells were detected after 24 h with the GST-CPAF.Conclusion The GST-CPAF from Chlamydophila pneumoniae can induce the secretion of proinflammatory cytokines TNF-α and IL-6 by human monocytic cells,and inhibited the proliferation of THP-1 cell and apoptosis in vitro.
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OBJECTIVE To investigate the distribution of clinical bacterial isolates and the change in antibiotic resistance spectrum in our hospital from 2005 to 2007.METHODS Data of bacterial susceptibility testing of clinical isolates from the Second Affiliated Hospital in of University of South China from 2005 to 2007 were collected and analyzed by software WHONET25.Results were assessed according to the National Committee for Clinical Laboratory Standards(NCCLS) of America issued in 2005.RESULTS The amount of Gram-negative bacteria decreased and of Gram-positive bacteria increased during this period.The proportion of coagulase negative Staphylococcus(CNS) had been increasing and reached 21.7% in 2007.The proportions of Staphylococcus aureus decreased from 17.6% in 2005 to 13.0% in 2007.Escherichia were the top two bacteria in 2007.The drug resistance rate of staphylococci against penicillin and erythromycin was more than 92.2% and 52.2%,respectively.The oxacillin resistance rate of CNS was 74.5%,significantly higher than that of S.aureus(16.5%).Drug resistance rate of Enterococcus to vancomycin was 1.1%.Gram-negative bacteria were found resistant to meropenem and imipenem.The resistance rate to ampicillin of Klebsiella and Escherichia was very high.CONCLUSIONS The variation of drug resistance and distribution of clinical bacterial isolates in our hospital are related to the improper use of antibiotics.It is very important to select antibiotics correctly according to the results of antibiotics susceptibility tests.
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Objective To review the research advances in molecular biology of vascular restenosis.Methods The literatures about molecular biology of vascular restenosis were reviewed.Results Current transgenic ways had some advantages and disadvantages. Gene therapy with HSV tk, Rb,p21,p27,p53,c myc, c myb, vascular endothelial growth factor,bFGF,platelet derived growth facfor,nuclear factor ?B and so on inhibited vascular restenosis.Conclusion A better transgenic system and gene combination therapy will be effective to treat vascular restenosis.
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Objective To evaluate surgical treatment of infected femoral artery pseudoaneurysm.Methods The data on surgical treatment of 45 patients with infected femoral artery pseudoaneurysm admitted from January 2003 to June 2008 were analyzed retrospectively.Fourty-three patients underwent operative treatment including excision of infected femoral artery pseudoaneurysm,exhaustive debridement and bypass graft with vascular prosthesis.Two patients were unavoidable to undergo removing of infected femoral artery pseudoaneurysm and ligating the proximal and distal artery of pseudoaneurysm because of severe infection and large volume.Results The patients were followed up from 3 to 12 months(mean 7.82 months).The limbs of all the patients underwent bypass graft with vascular prosthesis were salvaged successfully,patients of which had secondary wound healing and had not intermittent lameness.One of two patients performed ligation of artery was salvaged successfully but had severe intermittent lameness,another patient underwent high amputation above knee because of ischemic gangrene.ConclusionFor infected femoral artery pseudoaneurysm,the operative treatment including excision of infected femoral artery pseudoaneurysm,exhaustive debridement and bypass graft with vascular prosthesis is effective and safe.
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Objective To evaluate the therapeutic effects of pingyangmycin in treatment of body surface hemangioma in children.Methods The clinical data of 1 658 children patients with hemangioma on body surface in which pingyangmycin was injected between January 1997 and January 2008 were analyzed retrospectively.Results All 1 658 patients were observed for 6-12 months,with average of 10.83 months.The total effective rate was 97.09%.Compared among different types of hemangioma,total effective rate had significant difference(?2=203.12,P
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Objective To evaluate the indications of different hepatic blood flow occlusion in hepatectomy of cancer patients. Methods One hundred and fifty-six patients admitted between 1991-2001 underwent hepatectomy with hepatic blood flow occlusion in different ways, among them 48 cases underwent hepatic segmentectomy with segmental portal vein occlusion by a balloon catheter, 71 cases underwent hepatectomy with porta hepatis occlusion by Pringle method, 37 cases treated by hemihepatectomy or partial hepatectomy with hemihepatic occlusion. Results Intraoperative blood loss in patients using balloon catheter was smaller and postoperative liver function recovered faster compared with other ways of blood flow occlusion. Conclusion The preliminary result shows that hepatic segmentectomy with segmental portal vein occlusion by a balloon catheter is safe and useful technique for hepatectomy.
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Objective: To investigate extracelluar signal-regulated kinase(ERK) pathway and cytotoxic effect after block of this pathway after 5-aminolevulinic acid(ALA)-based photodynamic therapy(PDT) on SW480 cell.Methods: SW480 was divided into control group,light group,ALA group,ALA-PDT group and PD98059 group.Western blot was used to detect the expression of protein products and phosphorylation protein products of MEK and ERK1/2.Optical density value and survival rate of each group were obtained at different time by MTT.Results: ERK pathway of SW480 cell was activated and block of this pathway increased cytotoxic effect of SW480 cell after ALA-PDT.Conclusions: Activation of ERK pathway protects SW480 cell from ALA-PDT.Block of ERK pathway at different levels may become a new target,which will enhance cytotoxic effect of ALA-PDT on colon carcinoma.