In this study, bacteriophage of an antibiotic-resistant Escherchia coil strain isolated from feces of chicken was isolated. Its host range was determined by the method of spotting sample on monolayer agar, and its lys...In this study, bacteriophage of an antibiotic-resistant Escherchia coil strain isolated from feces of chicken was isolated. Its host range was determined by the method of spotting sample on monolayer agar, and its lysis titer, optimal multiplicity of infection （MOI）, temperature tolerance and pH tolerance were determined by the double-layer agar plate method. The results showed that the bacteriophage had a broad host range. The biological assay demonstrated that two strains of E. coil were fully lysed and one strain of E. coil was weakly lysed by the bacteriophage. The lysis titer and MOI of the bacteriophage were 1.20×10^8 PFU/ml and 1, respec- tively. Under the optimum temperature of 40℃, the Jysis titer of the bacteriophage reached 8.90×10^9 PFU/ml; however, the bacteriophage lost its infectivity at the tem- perature of 80℃. In the pH range of 5-11, the lysis titer of the bacteriophage ranged from10^6 to 10^9 PFU/mI. Under the condition of pH 4 and 12, the bacterio- phage was invalid.展开更多
Objective: To study the reversal effect of neferine on adriamycin (ADM) resistant human breast cancer cell line MCF-7/ADM. Methods: The cytotoxic effect of Nef or ADM was determined by 3-[4, 5-dimethylthiazol-2.-yl], ...Objective: To study the reversal effect of neferine on adriamycin (ADM) resistant human breast cancer cell line MCF-7/ADM. Methods: The cytotoxic effect of Nef or ADM was determined by 3-[4, 5-dimethylthiazol-2.-yl], 5-diphenyl tetraxolium bromid (MTT) assay. Apoptosis and the expression of P-glycoprotein (P-gp) were detected by flow cytometry (FCM). The intracellular ADM concentration was measured by HPLC. Results: Nef at 1, 5, 10 mol/L decreased the IC50 of ADM to MCF-7/ADM from 11.63 g/mL to 4.59, 2.44, 0.27 g/mL respectively. MCF-7/ADM could resist the apoptosis induced by ADM while Nef (1-10 mol/L) could augment ADR-mediated apoptosis. Nef (10 mol/L) increased the accumulation of ADM up to 2.88 fold in MCF-7/ADM but not in sensitive cells MCF-7/S and reduced the expression of P-gp in MCF-7/ADM cells. Conclusion: Nef can circumvent multidrug resistance (MDR) of MCF-7/ADM cells and the mechanism was associated with the increase of intracellular accumulation of ADM and the reduced expression of P-gp in MCF-7/ADM cells.展开更多
AIM： To isolate and identify the biological characteristics of human colon cancer stem cells （SW1116 cells） and further study their proteome. METHODS： SW1116 cells were isolated and cultured with a serum-free medi...AIM： To isolate and identify the biological characteristics of human colon cancer stem cells （SW1116 cells） and further study their proteome. METHODS： SW1116 cells were isolated and cultured with a serum-free medium （SFM）. Sphere formation was assayed to observe the formation of colon cancer stem cell spheres. SW1116 cells were inoculated into a serum-containing medium for observing their differentiation characteristics. Proliferation curve and cross-resistance of SWl116 cells to different drugs were detected by MTT. Percentage of SP cells in SW1116 cells was detected with Hoechst33342 staining. Telomerase activity in SW1116cells was checked by polymerase chain reaction （PCR）-enzyme linked immunosorbent assay. Expressions of stem cell relevant genes and proteins were detected by reverse transcription-PCR and Western blot, respectively. Total protein was isolated from SW1116 cells by two-dimensional gel electrophoresis （2-DE） and differentially expressed proteins were identified by tandem mass spectrometry （MALDI-TOF/TOF）. RESULTS： The isolated SW1116 cells presented as spheroid and suspension growths in SFM with a strong self-renewal, proliferation, differentiation and drug-resistance ability. The percentage of SP cells in SW1116 cells was 38.9%. The SW1116 cells co-expressed the CD133 and CD29 proteins. The telomerase activity in SW1116 cells was increased. The expressions of different stem cell relevant genes and proteins were detected. The proteomic analysis showed that the 26 protein spots were differently expressed in SW1116 cells and 10 protein spots were identified as ubiquitin fusion- degradation l-like protein, nuclear chloride channel protein, tubulin 13, Raichu404X, stratifin, F-actin cap- ping protein α-1 subunit, eukaryotic translation elongation factor 1 delta isoform 2, hypothetical protein, glyceraldehyde-3-phosphate dehydrogenase and guanine nucleotide binding protein 13 polypeptide 2-like 1, respectively. CONCLUSION： SW1116 cells are biologically characterized by self-renewal, proliferation and differentiation, and the differently expressed proteins in SW1116 cells may be essential for isolating cancer stem cells.展开更多
Primary hepatocellular carcinoma （HCC） is a quite frequent tumor which results in high mortality and most often exhibits a poor response to present drug therapies. Clearly, a thorough understanding of the biological...Primary hepatocellular carcinoma （HCC） is a quite frequent tumor which results in high mortality and most often exhibits a poor response to present drug therapies. Clearly, a thorough understanding of the biological bases of this malignancy might suggest new strategies for its treatment. Here we examine the evidences that both ＂pharmacological＂ mechanisms （e.g. drug transporter or detoxification enzyme over-expression） and alterations in other critical factors, including the IAPs （Inhibitory of Apoptosis Proteins）, involved in enhancement of cell survival and proliferation may determine the therapeutic resistance of HCC; we also underline the possible role in the process of the activation of transcription factors, like NF-κB, capable of contemporaneously up-regulating the mechanisms discussed. On this basis, we finally comment on the possible use of natural multi-targeted antitumoral agents like plant polyphenols to achieve sensitization to treatments in HCC.展开更多
Multidrug resistance （MDR） is a major problem in cancer chemotherapy. One of the best known mechanisms of MDR is the elevated expression of ATP-binding cassette （ABC） transporters. While some members of human ABC ...Multidrug resistance （MDR） is a major problem in cancer chemotherapy. One of the best known mechanisms of MDR is the elevated expression of ATP-binding cassette （ABC） transporters. While some members of human ABC transporters have been shown to cause drug resistance with elevated expression, it is not yet known whether the over-expression of other members could also contribute to drug resistance in many model cancer cell lines and clinics. The recent development ofmicroarrays and quantitative PCR arrays for expression profiling analysis of ABC transporters has helped address these issues. In this article, various arrays with limited or full list of ABC transporter genes and their use in identifying ABC transporter genes in drug resistance and chemo-sensitivity prediction will be reviewed.展开更多
AIM： To assess the safety of Bifidobacterium/ongum （B. longum） JDM301 based on complete genome sequences. METHODS： The complete genome sequences of JDM301 were determined using the GS 20 system. Putative virulence...AIM： To assess the safety of Bifidobacterium/ongum （B. longum） JDM301 based on complete genome sequences. METHODS： The complete genome sequences of JDM301 were determined using the GS 20 system. Putative virulence factors, putative antibiotic resis- tance genes and genes encoding enzymes respon- sible for harmful metabolites were identified by blast with virulence factors database, antibiotic resistance genes database and genes associated with harmful metabolites in previous reports. Minimum inhibitory concentration of 16 common antimicrobial agents was evaluated by E-test RESULTS： JDM301 was shown to contain 36 genes as- sociated with antibiotic resistance, 5 enzymes related to harmful metabolites and 162 nonspecific virulence fac- tors mainly associated with transcriptional regulation, adhesion, sugar and amino acid transport. B. longum JDM301 was intrinsically resistant to ciprofloxacin, ami- kacin, gentamicin and streptomycin and susceptible to vancomycin, amoxicillin, cephalothin, chloramphenicol, erythromycin, ampicillin, cefotaxime, rifampicin, imi- penem and trimethoprim-sulphamethoxazol. JDM301 was moderately resistant to bacitracin, while an earlier study showed that bifidobacteria were susceptible to this antibiotic. A tetracycline resistance gene with the risk of transfer was found in JDM301, which needs to be experimentally validated. CONCLUSION： The safety assessment of JDM301 using information derived from complete bacterial ge- nome will contribute to a wider and deeper insight into the safety of probiotic bacteria.展开更多
Enterococcus faecalis isolates （87） were phenotypically and genotypically identified and subsequently subjected to the antagonism test and antimicrobial susceptibility. The lipolitic, hemolytic and DNAse activities ...Enterococcus faecalis isolates （87） were phenotypically and genotypically identified and subsequently subjected to the antagonism test and antimicrobial susceptibility. The lipolitic, hemolytic and DNAse activities were identified along with the genes gelE, cylL, cylS, ccf, cpd and cob that, encode virulence determinants. Thirty seven percent of isolates inhibited Listeria monocytogenes （CERELA）, Listeria innocuous （CERELA）, Staphylococcus aureus （ATCC25932）, Lactococcus lactis （IL1403）, Micrococcus luteus （ATCC 10240） and Enterococcusfaecalis （ATCC29212）. All strains were sensitive to the ampicillin antibiotic, but 47% were resistant to at least one antimicrobial agent and 6% of isolates presented multidrug resistance. Ninety seven percent of isolates contained the gelE gene, but 77% of these isolates showed gelatinase activity. Presence of cylL and cylS genes was observed in 25% of the isolates, but only 5% presented hemolytic activity. None isolates showed lipase and DNAse activities. Eight percent of isolates contained the ccf gene and 2% showed the presence of the cpd and cob genes. The ability to inhibit pathogenic bacteria, low resistance to antibiotics and absence of virulence factors make some of Enterococcusfaecalis strains characterized in the present study promising for exploitation in other applications such as probiotics in aquaculture.展开更多
In the past decade,the advent of the epidermal growth factor receptor-tyrosine kinase inhibitors(EGFR-TKIs)has dramatically influenced the therapeutic strategies for treating lung cancer,but with tumor progression and...In the past decade,the advent of the epidermal growth factor receptor-tyrosine kinase inhibitors(EGFR-TKIs)has dramatically influenced the therapeutic strategies for treating lung cancer,but with tumor progression and drug resistance,patients will ultimately develop reduced sensitivity to EGFR-TKIs.How can we delay the emergence of drug resistance? What is the next strategy after drug resistance? How to reasonably combine platinum-based chemotherapy and EGFR-TKIs? These questions are currently the focus of lung cancer research.Clinical studies have reported that platinum-based chemotherapy can increase the sensitivity to EGFR-TKIs.However,results of pre-clinical and clinical studies have been inconsistent.The mechanisms of platinum chemotherapy and EGFR-TKIs are still unknown due to the lack of systematic research.Therefore,systematic studies are required to show the mechanisms of EGFR-TKIs and chemotherapy agents and define the markers sensitive to their combinations when given concurrently or sequentially.展开更多
基金Supported by Special Fund for Ocean and Fisheries Science Technology and Industrial Development of Guangdong Province(A201508A05)~~
文摘In this study, bacteriophage of an antibiotic-resistant Escherchia coil strain isolated from feces of chicken was isolated. Its host range was determined by the method of spotting sample on monolayer agar, and its lysis titer, optimal multiplicity of infection （MOI）, temperature tolerance and pH tolerance were determined by the double-layer agar plate method. The results showed that the bacteriophage had a broad host range. The biological assay demonstrated that two strains of E. coil were fully lysed and one strain of E. coil was weakly lysed by the bacteriophage. The lysis titer and MOI of the bacteriophage were 1.20×10^8 PFU/ml and 1, respec- tively. Under the optimum temperature of 40℃, the Jysis titer of the bacteriophage reached 8.90×10^9 PFU/ml; however, the bacteriophage lost its infectivity at the tem- perature of 80℃. In the pH range of 5-11, the lysis titer of the bacteriophage ranged from10^6 to 10^9 PFU/mI. Under the condition of pH 4 and 12, the bacterio- phage was invalid.
文摘Objective: To study the reversal effect of neferine on adriamycin (ADM) resistant human breast cancer cell line MCF-7/ADM. Methods: The cytotoxic effect of Nef or ADM was determined by 3-[4, 5-dimethylthiazol-2.-yl], 5-diphenyl tetraxolium bromid (MTT) assay. Apoptosis and the expression of P-glycoprotein (P-gp) were detected by flow cytometry (FCM). The intracellular ADM concentration was measured by HPLC. Results: Nef at 1, 5, 10 mol/L decreased the IC50 of ADM to MCF-7/ADM from 11.63 g/mL to 4.59, 2.44, 0.27 g/mL respectively. MCF-7/ADM could resist the apoptosis induced by ADM while Nef (1-10 mol/L) could augment ADR-mediated apoptosis. Nef (10 mol/L) increased the accumulation of ADM up to 2.88 fold in MCF-7/ADM but not in sensitive cells MCF-7/S and reduced the expression of P-gp in MCF-7/ADM cells. Conclusion: Nef can circumvent multidrug resistance (MDR) of MCF-7/ADM cells and the mechanism was associated with the increase of intracellular accumulation of ADM and the reduced expression of P-gp in MCF-7/ADM cells.
基金Supported by Medical Guidance Project of Shanghai Science Committee (No. 10411961800)Youth Science Fund of Fudan University (No. 08FQ49)
文摘AIM： To isolate and identify the biological characteristics of human colon cancer stem cells （SW1116 cells） and further study their proteome. METHODS： SW1116 cells were isolated and cultured with a serum-free medium （SFM）. Sphere formation was assayed to observe the formation of colon cancer stem cell spheres. SW1116 cells were inoculated into a serum-containing medium for observing their differentiation characteristics. Proliferation curve and cross-resistance of SWl116 cells to different drugs were detected by MTT. Percentage of SP cells in SW1116 cells was detected with Hoechst33342 staining. Telomerase activity in SW1116cells was checked by polymerase chain reaction （PCR）-enzyme linked immunosorbent assay. Expressions of stem cell relevant genes and proteins were detected by reverse transcription-PCR and Western blot, respectively. Total protein was isolated from SW1116 cells by two-dimensional gel electrophoresis （2-DE） and differentially expressed proteins were identified by tandem mass spectrometry （MALDI-TOF/TOF）. RESULTS： The isolated SW1116 cells presented as spheroid and suspension growths in SFM with a strong self-renewal, proliferation, differentiation and drug-resistance ability. The percentage of SP cells in SW1116 cells was 38.9%. The SW1116 cells co-expressed the CD133 and CD29 proteins. The telomerase activity in SW1116 cells was increased. The expressions of different stem cell relevant genes and proteins were detected. The proteomic analysis showed that the 26 protein spots were differently expressed in SW1116 cells and 10 protein spots were identified as ubiquitin fusion- degradation l-like protein, nuclear chloride channel protein, tubulin 13, Raichu404X, stratifin, F-actin cap- ping protein α-1 subunit, eukaryotic translation elongation factor 1 delta isoform 2, hypothetical protein, glyceraldehyde-3-phosphate dehydrogenase and guanine nucleotide binding protein 13 polypeptide 2-like 1, respectively. CONCLUSION： SW1116 cells are biologically characterized by self-renewal, proliferation and differentiation, and the differently expressed proteins in SW1116 cells may be essential for isolating cancer stem cells.
文摘Primary hepatocellular carcinoma （HCC） is a quite frequent tumor which results in high mortality and most often exhibits a poor response to present drug therapies. Clearly, a thorough understanding of the biological bases of this malignancy might suggest new strategies for its treatment. Here we examine the evidences that both ＂pharmacological＂ mechanisms （e.g. drug transporter or detoxification enzyme over-expression） and alterations in other critical factors, including the IAPs （Inhibitory of Apoptosis Proteins）, involved in enhancement of cell survival and proliferation may determine the therapeutic resistance of HCC; we also underline the possible role in the process of the activation of transcription factors, like NF-κB, capable of contemporaneously up-regulating the mechanisms discussed. On this basis, we finally comment on the possible use of natural multi-targeted antitumoral agents like plant polyphenols to achieve sensitization to treatments in HCC.
文摘Multidrug resistance （MDR） is a major problem in cancer chemotherapy. One of the best known mechanisms of MDR is the elevated expression of ATP-binding cassette （ABC） transporters. While some members of human ABC transporters have been shown to cause drug resistance with elevated expression, it is not yet known whether the over-expression of other members could also contribute to drug resistance in many model cancer cell lines and clinics. The recent development ofmicroarrays and quantitative PCR arrays for expression profiling analysis of ABC transporters has helped address these issues. In this article, various arrays with limited or full list of ABC transporter genes and their use in identifying ABC transporter genes in drug resistance and chemo-sensitivity prediction will be reviewed.
基金Supported by The National Key Program for Infectious Diseases of China,No. 2008ZX10004 and 2009ZX10004the Program of Shanghai Subject Chief Scientist,No. 09XD1402700+1 种基金the Program of Shanghai Research and Development,No. 10JC1408200a China Partnering Award from the Biotechnology and Biological Sciences Research Council,United Kingdom
文摘AIM： To assess the safety of Bifidobacterium/ongum （B. longum） JDM301 based on complete genome sequences. METHODS： The complete genome sequences of JDM301 were determined using the GS 20 system. Putative virulence factors, putative antibiotic resis- tance genes and genes encoding enzymes respon- sible for harmful metabolites were identified by blast with virulence factors database, antibiotic resistance genes database and genes associated with harmful metabolites in previous reports. Minimum inhibitory concentration of 16 common antimicrobial agents was evaluated by E-test RESULTS： JDM301 was shown to contain 36 genes as- sociated with antibiotic resistance, 5 enzymes related to harmful metabolites and 162 nonspecific virulence fac- tors mainly associated with transcriptional regulation, adhesion, sugar and amino acid transport. B. longum JDM301 was intrinsically resistant to ciprofloxacin, ami- kacin, gentamicin and streptomycin and susceptible to vancomycin, amoxicillin, cephalothin, chloramphenicol, erythromycin, ampicillin, cefotaxime, rifampicin, imi- penem and trimethoprim-sulphamethoxazol. JDM301 was moderately resistant to bacitracin, while an earlier study showed that bifidobacteria were susceptible to this antibiotic. A tetracycline resistance gene with the risk of transfer was found in JDM301, which needs to be experimentally validated. CONCLUSION： The safety assessment of JDM301 using information derived from complete bacterial ge- nome will contribute to a wider and deeper insight into the safety of probiotic bacteria.
文摘Enterococcus faecalis isolates （87） were phenotypically and genotypically identified and subsequently subjected to the antagonism test and antimicrobial susceptibility. The lipolitic, hemolytic and DNAse activities were identified along with the genes gelE, cylL, cylS, ccf, cpd and cob that, encode virulence determinants. Thirty seven percent of isolates inhibited Listeria monocytogenes （CERELA）, Listeria innocuous （CERELA）, Staphylococcus aureus （ATCC25932）, Lactococcus lactis （IL1403）, Micrococcus luteus （ATCC 10240） and Enterococcusfaecalis （ATCC29212）. All strains were sensitive to the ampicillin antibiotic, but 47% were resistant to at least one antimicrobial agent and 6% of isolates presented multidrug resistance. Ninety seven percent of isolates contained the gelE gene, but 77% of these isolates showed gelatinase activity. Presence of cylL and cylS genes was observed in 25% of the isolates, but only 5% presented hemolytic activity. None isolates showed lipase and DNAse activities. Eight percent of isolates contained the ccf gene and 2% showed the presence of the cpd and cob genes. The ability to inhibit pathogenic bacteria, low resistance to antibiotics and absence of virulence factors make some of Enterococcusfaecalis strains characterized in the present study promising for exploitation in other applications such as probiotics in aquaculture.
文摘In the past decade,the advent of the epidermal growth factor receptor-tyrosine kinase inhibitors(EGFR-TKIs)has dramatically influenced the therapeutic strategies for treating lung cancer,but with tumor progression and drug resistance,patients will ultimately develop reduced sensitivity to EGFR-TKIs.How can we delay the emergence of drug resistance? What is the next strategy after drug resistance? How to reasonably combine platinum-based chemotherapy and EGFR-TKIs? These questions are currently the focus of lung cancer research.Clinical studies have reported that platinum-based chemotherapy can increase the sensitivity to EGFR-TKIs.However,results of pre-clinical and clinical studies have been inconsistent.The mechanisms of platinum chemotherapy and EGFR-TKIs are still unknown due to the lack of systematic research.Therefore,systematic studies are required to show the mechanisms of EGFR-TKIs and chemotherapy agents and define the markers sensitive to their combinations when given concurrently or sequentially.
