Analysis on drug-resistance and molecular epidemiology of Acinetobacter baumannii isolated from the clinical samples in two Chinese hospitals

In the present study, the drug-resistance genes encodingβ-lactamases, aminoglycoside modifying enzymes, DNA topoisomerases and integron as well as their molecular epidemiology were investigated by means of analyzing the drug-resistance and molecular epidemiology of Acinebacter bau mannii isolated from the clinical samples in two hospitals in Qiangzhou and Huzhou city of Jiangsu and Zhejiang province from July 2000 to March 2005. The minimal inhibitory concentrations (MICs) of these 307 isolates were detected by automatic microbiological system, and 35 strains against 5-fluoro-quinolones were performed by agar dilution assay. Meanwhile, the resistant genes in 80 isolates were amplified by PCR with identification by DNA sequencer. It was found that most of the 307 isolates of A . baumannii were resistant to multiple antibiotics tested, in which the resistance rates of the isolates against piperacillin, piperacillin/tazobactam, amoxacillin/clavulanic acid, cefotaxime, ceftazidime, cefepime, gentamicin, amikacin, ciprofloxacin, chloramphenicol and sulfamethoxazole/trimethoprim were all above 35% , but those of imipenem and meropenem were quite low, ranged only 2.6% and 3.3%. In addition, it was also demonstrated that the positive rates of TEM and SHVβ-lactamase genes accounted for 93.8% and 22.5% respectively, and those of the aminoglycoside-modifying enzyme genes including aacC1, aacC2, aacC3, aacC4, aacC4A, aphA6, ant(2")-I and ant(3") I were 58.8%, 8.8%, 7.5%, 28.8%, 45.0%, 2.5%, 28.8% and 65.0% respectively. The mutations in the quinolone-resistant determining region (QRDR) of gyrA and parC genes indicated that substitution in Ser-83 residue of GyrA protein was most frequently occurred among strains with MIC for ciprofloxacin of more than 4μg/ml, whereas a double mutation at Ser-83 residue of gyrA and Ser-80 of parC was found in strains with MIC of ciprofloxacin of more than 8μg/ml. As to the positive rates of class 1 integron (Int I -1) and qacE△1-sul-1, it was found to be 60.0% and 77.5% respectively, and the rates of resistant genes of strains isolated in these two hospitals varied considerably. The results obtained in the present study indicate the presence of the multiple resistant genes in strains of A. baumannii , and great measures should be taken to control the spread of the resistant strains carrying the resistant genes.     

泛耐药肺炎克雷伯菌株的医院内流行特性的研究

目的 了解华山医院泛耐药肺炎克雷伯菌株及其流行的特点.方法 收集2006年8月-2009年12月对CLSI推荐常规检测药物均耐药的肺炎克雷们菌临床分离株,共57株.所有菌株都进行药物敏感试验、超广谱β-内酰胺酶(ESBLs)初筛及表型确证试验、改良Hodge试验、等电聚焦电泳,聚合酶链反应及其产物测序、接合试验、肠杆菌基因间重复共有序列PCR(ERIC-PCR)和多位点序列分型(MLST).结果 所有菌株都携带blaKPC-2、blaCTX-M-14、blaSHV12和blaTEM-1及qnrB和aac(6')-I b-cr基因.57株细菌中ST423型5株,MIST ST11型52株.ST423型散发,而ST11型呈医院内流行.57株细菌都对替加环素耐药,对多黏菌素、米诺环素和多西环素部分敏感.结论 本次泛耐药肺炎克雷们流行主要为ST11型菌株;不同的肺炎克雷伯菌株,播散能力不同;检出泛耐药肺炎克雷伯菌时应增加检测药物的种类.

Abstract：

Objective To understand the epidemic characteristics of an outbreak of panresistance Klebsiella pneumoniae occurred between 2006 and 2009 in a university hospital of Shanghai, China. Methods A total of 57 panresistance K. pneumoniae isolates were collected from August 2006 to December 2009.Antibiotic susceptibility of the isolates were determined by Kirby-Bauer disc diffusion method and microbroth dilution (MBD). ESBLs-producing initial screen test and phenotypic confirmatory test and carbapenemase-producing modified Hodge test ( MHT) were performed to detect the resistance phenotype of the isolates. Be-ta-lactamases were studied by IEF, PCR and the product sequencing. While conjugation assay were conducted to understand the transferability of these genes. The genetic relationship between isolates was established by ERIC-PCR and multilocus sequence typing (MLST). Except for the antibiotics recommended by CLSI guideline in the routine test, the other antibiotics were added to find out the effective drugs to treat the infection. Results All 57 isolates were highly resistant to all examined antibiotics. All isolates produced ESBLs and carbapenemase. IEF revealed that each isolate produced four beta-lactamases. All isolates carried blaKPC-2,blaCTX-M-14,blaSHV12,blaTEM-1,qnrB and aac(6') - I b-cr. Forty-four of the 57 (77.2% ) isolates were successful to transfer their resistance genes to E. coli recipient J53 by conjugation assay. By RAPD, all 57 isolates were grouped into two genotypes that were further identified as members of MUST types 423 and 11.Sequence types 423(ST423) only occurred before May 2008 and ST11 occurred (52 isolates) after May 2008. Most of isolates of the outbreak were ST11 (91. 2% ). A part of isolates were susceptive to added antibiotics. Conclusion The outbreak of panresistance K.pneumoniae was caused by those isolates which carried multiple resistant genes. There is a different ability of dissemination between different ST types K. pneumoniae isolate. It was necessary to add the antibiotics to find out the effective drugs to treat the infection.     

Molecular mechanism of the qnrA genemediated quionlone resistance in Gram-negative bacteria

To explore the prevalence of the plasmid-mediated quinolone resistance gene qnrA in Gramnegative bacteria and to investigate its molecular genetic background and resistance profile in isolates harboring this gene, a total of 629 nalidixic acid-resistant isolates of non-repetitive Gram-negative bacteria were collected from clinical specimens between April 2004 and April 2006 and these isolates were screened for qnrA gene by PCR using specific primers combined with DNA sequencing. The extended spectrum β-1actamase （ESBL） or AmpC-producing isolates were distinguished by the phenotypic confirmatory test combined with DNA sequencing, and the antibiotics susceptibility test for qnrA-positive isolates was carried out by Kirby-Bauer and E-test method. To detect the location of the qnrA gene, plasmid conjugation and Southern hybridization were performed and the integron structure containing the qnrA gene was cloned by PCR strategy and sequenced by primer walking. It was demonstrated that the incidence of the qnrA-positive strains in nalidixic acid-resistant bacteria was 1.9% （12/629）, in which the detection rates for Klebiesiella pneumoniae. Enterobacter cloacae, Enterobacter aerogenes, Citrobacterfreundii and Salmonella choeraesuis were 2.2% （3/138）, 17. 1% （6/35）, 9. 1% （1/11）, 12.5% （1/8）, and 14.3% （1/7）, respectively. The qnrA gene was found to be embedded in the complex sull-type integron located on plasmids with varied size （80-180 kb）. Among them, 4 qnrA-positive isolates carried integron In37 and 8 isolates carried a novel integron, temporarily desig- nated as InX. All the qnrA-positive isolates were ESBL-producing and transferable for the multi-drug resistance. It is concluded that the plasmid-mediated drug-resistance mechanism exists in the quinolone resistant strains of isolates from hospitals in Guangdong area, but the incidence was rather low. Nevertheless, it is still possible that the horizontal transfer of the resistant qnrA gene might lead to the spreading of drug-resista     

Preparation and application of polyclonal antibody against a recombinant laccase

A laccase gene from Trametes sp. 420 was recombinantly expressed in Pichia pastoris, producing the enzyme rLacD. Six mutant enzymes were produced by site-directed mutation at six potential glycosylation sites in the enzyme rLacD respectively. To probe the mutants with lower activities sensitively and specifically, the antiserum containing specific polyclonal antibodies were prepared by immunizing healthy male rabbits, about 4-month-old and 2 kilogram weight, using pure rLacD as an immunogen. Antibodies were collected after the fifth immunization injection. The antiserum had titres of 1：32 in double immunodiffusion test and of 1：128,000 in enzyme-linked immunosorbent assay （ELISA）. The results obtained by Western blot analysis showed that the antiserum could react with rLacD and its mutants with highly specific and sensitive affinities.     

广州地区携带blaCTX-M-15流行质粒的分子特征研究

目的 研究广州地区携带blaCTX-M-15质粒的分子特征.方法 以2007年到2008年期间来源于广州9家医院确证产CTX-M-15 ESBL的38株大肠埃希菌和47株肺炎克雷伯菌为研究对象,脉冲场凝胶电泳(PFGE)和多位点序列分型(MLST)法检测blaCTX-M-15阳性株的同源性,MIC法检测菌株对常用抗生素的敏感性.血清凝集试验确定大肠埃希菌血清型.接合试验了解携带blaCTX-M-15质粒的可接合性,限制性内切酶分析质粒的同源性.PCR分析大肠埃希菌的系统发育群,流行质粒的不相容群和耐药基因构成.结果 blaCTX-M-15阳性的大肠埃希菌和肺炎克雷伯菌分别用PFGE确定为28个和30个基因型.携blaCTX-M-15大肠埃希菌的系统发育群以D群(67.8%)和A群(21.4%)为主.大肠埃希菌血清型呈散在分布,未发现O25:H4血清型.大肠埃希菌和肺炎克雷伯菌的ST型无明显集中趋势,大肠埃希菌中发现非O25血清型的ST131,肺炎克雷伯菌中发现2个新的tonB基因和2个新的ST型.58个独立克隆的代表菌株中有40株可将携blaCTX-M-15的质粒传递给受体菌E.coli C600(Rif),其中大肠埃希菌有19株,肺炎克雷伯菌有21株.57.9%(11/19)的大肠埃希菌中携带blaCTX-M-15的质粒大小为65 kb,85.7%(18/21)的肺炎克雷伯菌中携带blaCTX-M-15的质粒大小为92 kb.限制性内切酶分析显示,这2种质粒属流行质粒,分别命名为p15-e和p15-k.通过PCR调查,p15-e存在blaCTX-M-15和ISEcpI;p15-k上存在blaCTX-M-15、ISEcpI、aac(6')-Ⅰ b、aac(3')-Ⅲ、blaOXA-1、qnrB、qnrS、blaDHA-1、blaTEM-1.p15-k同时被证实属于质粒不相容群FⅡ群.结论 广州地区存在携带blaCTX-M-15流行质粒的传播,且流行质粒的大小和基因结构与国外报道不同;无克隆株传播证据.

Abstract：

Objective To study the molecular characteristic of the epidemic plasmids carrying blaCTX-M-15 in Guangzhou. Methods A total of 38 strains of E. coli and 47 strains of K. pneumoniae both producing CTX-M-15 ESBLs were collected from nine hospitals in Guangzhou from 2007 to 2008. The clonal relationship of isolates carrying blaCTX-M-15 was determined by PFGE and MLST. Antimicrobial susceptibility was determined by microdilution test for all isolates. Conjugative plasmids carrying blaCTX-M-15 were obtained by mating and were subject to restriction analysis. PCR was used to determine phylogenetic groups of E. coli,and to study replicon type and the genetic contexts of the plasmids harboring blaCTX-M-15. Serum agglutination test was used to detect the serotype of E. coli. Results The 37 strains of E. coli were classified into 28 genotypes, while the 47 strains of K. pneumoniae were divided into 30 genotypes. ST131 was found in E. coli but not O25 serotype. Two novel-alleles of tonB and new ST were determined in K. pneumoniae. Forty out of 58 isolates represented independent genotypes have been succeeded to transfer the plasmid carrying blaCTX-M-15 to the E. coli C600(Rif) by conjugation. The sizes of plasmids carrying blaCTX-M-15 are 65 kb in 57.9% isolates of E. coli and 92 kb in 87.5% isolates of K. pneumoniae. Two epidemic plasmids were detected in E.coli and K. pneumoniae by restriction enzyme, designated p15-e and p15-k respectively. The blaCTX-M-15 and ISEcpI were identified on p15-e, and the blaCTX-M-15 ,ISEcpI,aac(6')- Ⅰ b,aac(3')-Ⅲ ,blaOXA-1 ,qnrB,qnrS,blaDHA-1 , blaTEM-1 were determined on p15-k. The p15-k also was identified to belong to the incompatible group FⅡ. Conclusion The local dissemination of blaCTX-M-15 appears to be due to the spread of epidemic plasmids harboring blaCTX-M-15. No evidence supports the dissemination of clone strains which carried blaCTX-M-15.     

肺炎克雷伯菌的喹诺酮外排基因qepA的检测及菌株同源性分析

目的 检测质粒介导的喹诺酮类外排基因qepA在临床分离的产超广谱β-内酰胺酶(ESBLs)的肺炎克雷伯菌中的分布情况,并对阳性菌株进行同源性分析.方法 41株细菌的鉴定和药敏采用Vitek-2 Compact系统;采用PCR法检测qepA并进行序列测定、比对.应用Rep-PCR法采用Diversilab系统对qepA阳性菌株进行同源性分析;同时了解阳性株上的rmtB分布情况.结果 5株肺炎克雷伯菌上检测到qepA基因,检出率为12.2%.Diversilab分析结果表明其中两株的qepA阳性菌同源性超过99%.包含qepA的肺炎克雷伯菌中2株(40%)检出rmtB.结论 在肺炎克雷伯菌临床菌株上检出质粒介导的喹诺酮类外排基因qepA.

Abstract：

Objective To investigate the prevalence of qepA, quinolone efflux protein, among 41 unique clinical strains of K. pneumoniae producing ESBLs and to study the qepA-bearing isolates using the Diversilab system. Methods Identification and antimicrobial susceptibility were performed by Vitek-2 Compact System. Screening of qepA was carried out by PCR amplification. The NCBI BLAST program was utilized for sequence comparisons. qnr-bearing strains was evaluated by the Repetitive-sequence-based PCR(Rep-PCR) employing the Diversilab system. And the existence of rmtB was detected among these qepA contained isolates. Results qepA were detected in 5 isolates( 12.2% ). The Rep-PCR profiles produced by the Diversilab system showed that 2/5 of isolates were indistinguishable. And 60% of qepA-positive isolates were detected to harbor rmtB gene. Conclusion The data suggest the emergence of qepA-borne K. pneumoniae.     

Correlation of the antiseptic resistance in Acinetobacter baumannii and the antiseptic resistance gene qacE Δ 1 located in class Ⅰ integron

In the past decade, uses of antiseptics and disinfectants in hospitals and other health care centers are rather common, but the chance to develop resistance to antiseptics and disinfectants is also increased. Acinetobacter baumannii is one of the opportunistic bacteria involving in the nosocomial infection . In the present study, the correlation of the antiseptic resistance in A . baumannii and the antiseptic resistance gene qacEΔ1 was investigated by means of determination of MICs. Meanwhile, the MICs of glutaraldehyde, chlorhexidine, benzalkonium bromide, iodophor and trichloroisocyanurate to 80 clinical isolates of A. baumannii were detected by tube dilution assay and the resistance genes intI1 and qacEΔ1 in these isolates were amplified by PCR and verified by DNA sequencer. It was found that the MIC50 for these 5 antiseptics tested were 32, 8, 8, 4 and 1μg/ml respectively, and the detection rates of intI1 and qacEΔ1 gene were 60.0% and 77.6% respectively. In addition, 55% of the 80 isolates simultaneously possessed both intll and qacEΔ1 gene, and the percentage of antiseptic resistance of A . baumannii carring both genes to benzalkonium bromide were higher than that without these two genes, however, there was no significant difference between intll and qacEΔ1 gene. The result in bactericidal efficiency assay indicated that chlorhexidine could still produce rapid and strong bactericidal effect at concentration of 1 MIC after 10 min exposure. These results suggest that the antiseptic resistance of A . baumannii to various antiseptics is correlated with the presence of the antiseptic resistance genes qacEΔ1 in bacteria, thus warning that the increase of the antiseptic resistance should not be ignored and the relative high concentration or prolonged application time is required to achieve a sufficient bactericidal effect.     

Correlation between expression of DcR3 on tumor cells and sensitivity to FasL

To investigate the correlation between sensitivity to Fas ligand （FasL） and expression level of decoy receptor 3 （DcR3） on tumor cell surface, Fas/DcR3 mRNA expression was detected by RT-PCR. Anti-DcR3 mAb was used to detect expression level of DcR3 on surface of tumor cells by flow cytometry. Caspase-8, caspase-9, caspase-3, Bcl-2 expressions were analyzed by Western blot, respectively. Sensitivity to apoptosis induced by FasL was determined by Annexin V apoptosis kit. The expressions of DcR3 on the surface of tumor cells from high to low were approximately 35.3% in BGC823 cells, and 21.6% in MCF-7 cells, respectively. The apoptotic rates induced by FasL from low to high were 15.6% in BGC823 cells, and 58.2% in MCF-7 cells, respectively. There was a significant correlation between the expression levels of DcR3 with FasL-inducing apoptosis. Cellular ＆ Molecular Immunology.     

Interleukin 17-Producing γδ T Cells Increased in Patients with Active Pulmonary Tuberculosis

Although it has been known that γδ T cells may play an important role in the immune response to infection of Mycobacterium tuberculosis （M. tb）, the mechanisms by which the γδ T cells participate in the innate and/or acquired immunity to tuberculosis （TB） have not been full elucidated. In the present study, 27 patients with active pulmonary TB and 16 healthy donors （HD） were performed. We found that proportion of IL-17-producing cells among lymphocyte was similar between TB patients and HD, whereas the proportions of γδ T cells in IL-17-producing cells （59.2%） and IL-17-producing cells in γδ T cells （19.4%） in peripheral blood were markedly increased in TB patients when compared to those in HD （43.9% and 7.7%, respectively）. In addition, the proportions of IFN-T-producing γδ T cells in TB patients were obviously lower than that in HD. Upon re-stimulated with M. tb heat-treated antigen （M. tb-HAg） in vitro, fewer IL-17-producing γδ T cells were generated from HD and TB patients, whereas IFN-T-producing γδ T cells were increased in TB patients compared to that in HD. Our findings in TB patients and healthy human were consistent with other murine investigation that the IL-17- producing γδ T cells were main source of IL-17 in mouse model of BCG infection, suggesting that γδ T cells might be involved in the formation of tubercular granuloma in pulmonary TB patients, but need further identification.     

Adherence of uropathogenic Escherichia coli to human primary epithelial cells of renal pelvis

Human primary epithelial cells of renal pelvis was established to investigate the adherence of uropathogenic Escherichia coli (UPEC) to this cell line, in which the primary cell culture was performed by using cultivation of the normal epithelium of renal pelvis in keratinocyte serum free medium (K-SFM) with epidermal growth factor (EGF) and bovine pituitary extract (BPE). Both UPEC132 obtained from urine specimen of patients with pyelonephritis and the pilus-free representative strain E. coli K-12p678-54 were used to study the adherence of these strains on human primary epithelial cells of renal pelvis. The UPEC adherence was performed with observation on the morphological changes of the adhered cells, while the adhesion rates and indices were calculated in different times of experiment. In addition, the virulence genes hly and cnf1 of UPEC132 were detected by multiplex PCR assay. In this study, the human primary epithelial cells of renal pelvis was found to exhibit the character of the transitional epithelial cells. Compared with the control group, the adhesion rates and indices began to increase from 15 min of the experiment time and reached its peak in 120 min. The adhesion rate and index of UPEC132 to human primary epithelial cells of renal pelvis were 74.4% and 34.0 respectively. Many microscopic changes in the primary cells adhered with UPEC132 could be detected, such as rounding or irregularity in shape, unevenness in staining and the cytoplasmic and nuclear changes. It suggests that human primary epithelial cells of renal pelvis can be used for the experiment on UPEC adhesion, thus providing a basis for the further study on the pathogenesis of UPEC.     

Construction of cDNA library from NPC tissue and screening of antigenic genes

To construct cDNA library of nasopharyngeal carcinoma (NPC) and obtain the NPC associated or specific antigens from it, we used a powerful new method to identify the antigens eliciting humoral immune response, which is SEREX (serological identification of antigen by recombinant cDNA expression library). Autologous serum of NPC patient was used to screen the reactive clones in the human NPC tissue cDNA library consisted of 3.64×106 recombinants. The 23 exact positive clones were subcloned to monoclonality and the size of cDNA inserts was identified by PCR. Then the nucleotide sequence of cDNA inserts was determined, and the sequence alignments were performed with BLAST software on GenBank database. They represented 16 different antigens. A detailed sequence analysis showed that 10 of 16 genes were high homologous to genes known in GenBank, such as RPL31, S100 A2, MT2A, etc. However, there were also 6 genes with low homology to genes in GenBank. Furthermore, 3 of 6 genes may be novel genes. The associations of these genes to NPC and the roles that they played in the occurrence and development of NPC should be further revealed.     

Analysis of genetic markers of N. Gonorrhoeae resistance to β-lactam antibiotics

A complex method for detection of genetic markers of N. gonorrhoeae resistance to penicillin was developed. Mutations in penA and ponA genes were detected by minisequencing reaction with subsequent detection of reaction products by MALDI-TOF mass spectrometry. This approach was tested on 31 clinical strains of N. gonorrhoeae with minimum inhibitory concentration of penicillin from 0.03 to 8 μg/ml and higher. Mutations in penA and ponA genes in moderately resistant strains were shown (minimum inhibitory concentration up to 0.5 μg/ml) and mutations in penA, ponA, and penB genes in resistant strains (minimum inhibitory concentration more than 1.0 μg/ml). β-Lactamase genes were detected in 4 strains with high resistance (minimum inhibitory concentration 4–8 and more μg/ml). Correlation between microbiological resistance and presence of respective mutations in the studied locuses was detected. __________ Translated from Byulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 141, No. 5, pp. 549–554, May, 2006     

Comparison of Ceramide Production with PLC Irradiated by 810 nm LED and 808 nm Laser

Due to its major role in maintaining the water-retaining properties of the epidermis, ceramide is of great commercial potential in cosmetics and pharmaceuticals such as hair and skin care products. However, current synthetic approaches for ceramide are tedious and time-consuming for industrial applications. Therefore, it is desirable to find an alternative cost-efficient and highly-yield method for obtaining the valuable products. In present study, the potential of producing ceramide through enzymatic hydrolysis of sphingomyelin with enzyme irradiated by 808 nm light has been studied. With enzyme irradiated by 808 nm light, its activity can be enhanced and reaction speed can be increased significantly.     

Expression of hypoxia inducible factor-1α and basic fibroblast factor in the tissues of colorectal adenocarcinoma

Objective To observe expression of hypoxia inducible factor-1α(HIF-1α)and basic fibroblast growth factor(BFGF)in the tissues of colorectal adenoearcinoma and analyze the relationship between expression of the two factors and biological behavior of colorectal adenocarcinoma.Methods The samples of colorectal adenocarcinoma(n=60)and para-adenocarcinoma(n=60)were taken from surgical resection patients and normal colorectal tissues (n=20) from patients with irritable bowel syndrome.The expression of HIF-1α and BFGF were detected by immunohistochemical staining (SP method).Results Positive rates of HIF-1αand BFGF in colorectal adenocarcinoma tissues were 61.7% and 65.0%,and positive rates of HIF-1α and BFGF in para-adenocarcinoma tissues were 10.0%and 13.3%(P<0.05,respectively);HIF-1αand BFGF were not detected in normal intestinal mucosa.There was no significant correlation with HIF-1α and BFGF expression in colorectal adenocarcinoma tissues to the sexuality,age,tumor size,tumorposition and differentiation(P>0.05).A significant correlation between expression of the two factors and Dukes stage was observed (P<0.05).Positive rates of HIF-1α and BFGF in colorectal adenocarcinoma tissuesof Dukes A and B stage were 47.2%and 52.7%,respectively.Positive rates of HIF-1α and BFGF in colorectal adenocarcinoma tissues of Dukes C and D stage were 83.3%and 83.3%.respectively.There was a positive correlation between expression of the two factors and carcinogenesis of colorectal adenocarcinoma(r=0.4276.P<0.001).Conclusion The results showed that the enhanced expression of HIF-1 α and BFGF incolorectal adenocarcinoma tissues may be associated with the prognosis of the patients with colorectal adenocarcinoma,and HIF-1α and BFGF may participate synergistically in the carcinogenesis of colorectal adenocarcinoma.     

Expression of hypoxia inducible factor-1α and basic fibroblast factor in the tissues of colorectal adenocarcinoma

Objective To observe expression of hypoxia inducible factor-1α(HIF-1α)and basic fibroblast growth factor(BFGF)in the tissues of colorectal adenoearcinoma and analyze the relationship between expression of the two factors and biological behavior of colorectal adenocarcinoma.Methods The samples of colorectal adenocarcinoma(n=60)and para-adenocarcinoma(n=60)were taken from surgical resection patients and normal colorectal tissues (n=20) from patients with irritable bowel syndrome.The expression of HIF-1α and BFGF were detected by immunohistochemical staining (SP method).Results Positive rates of HIF-1αand BFGF in colorectal adenocarcinoma tissues were 61.7% and 65.0%,and positive rates of HIF-1α and BFGF in para-adenocarcinoma tissues were 10.0%and 13.3%(P<0.05,respectively);HIF-1αand BFGF were not detected in normal intestinal mucosa.There was no significant correlation with HIF-1α and BFGF expression in colorectal adenocarcinoma tissues to the sexuality,age,tumor size,tumorposition and differentiation(P>0.05).A significant correlation between expression of the two factors and Dukes stage was observed (P<0.05).Positive rates of HIF-1α and BFGF in colorectal adenocarcinoma tissuesof Dukes A and B stage were 47.2%and 52.7%,respectively.Positive rates of HIF-1α and BFGF in colorectal adenocarcinoma tissues of Dukes C and D stage were 83.3%and 83.3%.respectively.There was a positive correlation between expression of the two factors and carcinogenesis of colorectal adenocarcinoma(r=0.4276.P<0.001).Conclusion The results showed that the enhanced expression of HIF-1 α and BFGF incolorectal adenocarcinoma tissues may be associated with the prognosis of the patients with colorectal adenocarcinoma,and HIF-1α and BFGF may participate synergistically in the carcinogenesis of colorectal adenocarcinoma.     

Expression of hypoxia inducible factor-1α and basic fibroblast factor in the tissues of colorectal adenocarcinoma

Objective To observe expression of hypoxia inducible factor-1α(HIF-1α)and basic fibroblast growth factor(BFGF)in the tissues of colorectal adenoearcinoma and analyze the relationship between expression of the two factors and biological behavior of colorectal adenocarcinoma.Methods The samples of colorectal adenocarcinoma(n=60)and para-adenocarcinoma(n=60)were taken from surgical resection patients and normal colorectal tissues (n=20) from patients with irritable bowel syndrome.The expression of HIF-1α and BFGF were detected by immunohistochemical staining (SP method).Results Positive rates of HIF-1αand BFGF in colorectal adenocarcinoma tissues were 61.7% and 65.0%,and positive rates of HIF-1α and BFGF in para-adenocarcinoma tissues were 10.0%and 13.3%(P<0.05,respectively);HIF-1αand BFGF were not detected in normal intestinal mucosa.There was no significant correlation with HIF-1α and BFGF expression in colorectal adenocarcinoma tissues to the sexuality,age,tumor size,tumorposition and differentiation(P>0.05).A significant correlation between expression of the two factors and Dukes stage was observed (P<0.05).Positive rates of HIF-1α and BFGF in colorectal adenocarcinoma tissuesof Dukes A and B stage were 47.2%and 52.7%,respectively.Positive rates of HIF-1α and BFGF in colorectal adenocarcinoma tissues of Dukes C and D stage were 83.3%and 83.3%.respectively.There was a positive correlation between expression of the two factors and carcinogenesis of colorectal adenocarcinoma(r=0.4276.P<0.001).Conclusion The results showed that the enhanced expression of HIF-1 α and BFGF incolorectal adenocarcinoma tissues may be associated with the prognosis of the patients with colorectal adenocarcinoma,and HIF-1α and BFGF may participate synergistically in the carcinogenesis of colorectal adenocarcinoma.     

Expression of hypoxia inducible factor-1α and basic fibroblast factor in the tissues of colorectal adenocarcinoma

Objective To observe expression of hypoxia inducible factor-1α(HIF-1α)and basic fibroblast growth factor(BFGF)in the tissues of colorectal adenoearcinoma and analyze the relationship between expression of the two factors and biological behavior of colorectal adenocarcinoma.Methods The samples of colorectal adenocarcinoma(n=60)and para-adenocarcinoma(n=60)were taken from surgical resection patients and normal colorectal tissues (n=20) from patients with irritable bowel syndrome.The expression of HIF-1α and BFGF were detected by immunohistochemical staining (SP method).Results Positive rates of HIF-1αand BFGF in colorectal adenocarcinoma tissues were 61.7% and 65.0%,and positive rates of HIF-1α and BFGF in para-adenocarcinoma tissues were 10.0%and 13.3%(P<0.05,respectively);HIF-1αand BFGF were not detected in normal intestinal mucosa.There was no significant correlation with HIF-1α and BFGF expression in colorectal adenocarcinoma tissues to the sexuality,age,tumor size,tumorposition and differentiation(P>0.05).A significant correlation between expression of the two factors and Dukes stage was observed (P<0.05).Positive rates of HIF-1α and BFGF in colorectal adenocarcinoma tissuesof Dukes A and B stage were 47.2%and 52.7%,respectively.Positive rates of HIF-1α and BFGF in colorectal adenocarcinoma tissues of Dukes C and D stage were 83.3%and 83.3%.respectively.There was a positive correlation between expression of the two factors and carcinogenesis of colorectal adenocarcinoma(r=0.4276.P<0.001).Conclusion The results showed that the enhanced expression of HIF-1 α and BFGF incolorectal adenocarcinoma tissues may be associated with the prognosis of the patients with colorectal adenocarcinoma,and HIF-1α and BFGF may participate synergistically in the carcinogenesis of colorectal adenocarcinoma.     

Expression of hypoxia inducible factor-1α and basic fibroblast factor in the tissues of colorectal adenocarcinoma

Objective To observe expression of hypoxia inducible factor-1α(HIF-1α)and basic fibroblast growth factor(BFGF)in the tissues of colorectal adenoearcinoma and analyze the relationship between expression of the two factors and biological behavior of colorectal adenocarcinoma.Methods The samples of colorectal adenocarcinoma(n=60)and para-adenocarcinoma(n=60)were taken from surgical resection patients and normal colorectal tissues (n=20) from patients with irritable bowel syndrome.The expression of HIF-1α and BFGF were detected by immunohistochemical staining (SP method).Results Positive rates of HIF-1αand BFGF in colorectal adenocarcinoma tissues were 61.7% and 65.0%,and positive rates of HIF-1α and BFGF in para-adenocarcinoma tissues were 10.0%and 13.3%(P<0.05,respectively);HIF-1αand BFGF were not detected in normal intestinal mucosa.There was no significant correlation with HIF-1α and BFGF expression in colorectal adenocarcinoma tissues to the sexuality,age,tumor size,tumorposition and differentiation(P>0.05).A significant correlation between expression of the two factors and Dukes stage was observed (P<0.05).Positive rates of HIF-1α and BFGF in colorectal adenocarcinoma tissuesof Dukes A and B stage were 47.2%and 52.7%,respectively.Positive rates of HIF-1α and BFGF in colorectal adenocarcinoma tissues of Dukes C and D stage were 83.3%and 83.3%.respectively.There was a positive correlation between expression of the two factors and carcinogenesis of colorectal adenocarcinoma(r=0.4276.P<0.001).Conclusion The results showed that the enhanced expression of HIF-1 α and BFGF incolorectal adenocarcinoma tissues may be associated with the prognosis of the patients with colorectal adenocarcinoma,and HIF-1α and BFGF may participate synergistically in the carcinogenesis of colorectal adenocarcinoma.     

Expression of hypoxia inducible factor-1α and basic fibroblast factor in the tissues of colorectal adenocarcinoma

Objective To observe expression of hypoxia inducible factor-1α(HIF-1α)and basic fibroblast growth factor(BFGF)in the tissues of colorectal adenoearcinoma and analyze the relationship between expression of the two factors and biological behavior of colorectal adenocarcinoma.Methods The samples of colorectal adenocarcinoma(n=60)and para-adenocarcinoma(n=60)were taken from surgical resection patients and normal colorectal tissues (n=20) from patients with irritable bowel syndrome.The expression of HIF-1α and BFGF were detected by immunohistochemical staining (SP method).Results Positive rates of HIF-1αand BFGF in colorectal adenocarcinoma tissues were 61.7% and 65.0%,and positive rates of HIF-1α and BFGF in para-adenocarcinoma tissues were 10.0%and 13.3%(P<0.05,respectively);HIF-1αand BFGF were not detected in normal intestinal mucosa.There was no significant correlation with HIF-1α and BFGF expression in colorectal adenocarcinoma tissues to the sexuality,age,tumor size,tumorposition and differentiation(P>0.05).A significant correlation between expression of the two factors and Dukes stage was observed (P<0.05).Positive rates of HIF-1α and BFGF in colorectal adenocarcinoma tissuesof Dukes A and B stage were 47.2%and 52.7%,respectively.Positive rates of HIF-1α and BFGF in colorectal adenocarcinoma tissues of Dukes C and D stage were 83.3%and 83.3%.respectively.There was a positive correlation between expression of the two factors and carcinogenesis of colorectal adenocarcinoma(r=0.4276.P<0.001).Conclusion The results showed that the enhanced expression of HIF-1 α and BFGF incolorectal adenocarcinoma tissues may be associated with the prognosis of the patients with colorectal adenocarcinoma,and HIF-1α and BFGF may participate synergistically in the carcinogenesis of colorectal adenocarcinoma.     

Expression of hypoxia inducible factor-1α and basic fibroblast factor in the tissues of colorectal adenocarcinoma

Objective To observe expression of hypoxia inducible factor-1α(HIF-1α)and basic fibroblast growth factor(BFGF)in the tissues of colorectal adenoearcinoma and analyze the relationship between expression of the two factors and biological behavior of colorectal adenocarcinoma.Methods The samples of colorectal adenocarcinoma(n=60)and para-adenocarcinoma(n=60)were taken from surgical resection patients and normal colorectal tissues (n=20) from patients with irritable bowel syndrome.The expression of HIF-1α and BFGF were detected by immunohistochemical staining (SP method).Results Positive rates of HIF-1αand BFGF in colorectal adenocarcinoma tissues were 61.7% and 65.0%,and positive rates of HIF-1α and BFGF in para-adenocarcinoma tissues were 10.0%and 13.3%(P<0.05,respectively);HIF-1αand BFGF were not detected in normal intestinal mucosa.There was no significant correlation with HIF-1α and BFGF expression in colorectal adenocarcinoma tissues to the sexuality,age,tumor size,tumorposition and differentiation(P>0.05).A significant correlation between expression of the two factors and Dukes stage was observed (P<0.05).Positive rates of HIF-1α and BFGF in colorectal adenocarcinoma tissuesof Dukes A and B stage were 47.2%and 52.7%,respectively.Positive rates of HIF-1α and BFGF in colorectal adenocarcinoma tissues of Dukes C and D stage were 83.3%and 83.3%.respectively.There was a positive correlation between expression of the two factors and carcinogenesis of colorectal adenocarcinoma(r=0.4276.P<0.001).Conclusion The results showed that the enhanced expression of HIF-1 α and BFGF incolorectal adenocarcinoma tissues may be associated with the prognosis of the patients with colorectal adenocarcinoma,and HIF-1α and BFGF may participate synergistically in the carcinogenesis of colorectal adenocarcinoma.