During infectious disease episodes, pathogens express distinct subsets of virulence factors which allow them to adapt to different environments. Hence, genes that are expressed or upregulated in vivo are implicated in...During infectious disease episodes, pathogens express distinct subsets of virulence factors which allow them to adapt to different environments. Hence, genes that are expressed or upregulated in vivo are implicated in pathogenesis. We used in vivo induced antigen technology (IVIAT) to identify antigens which are expressed during infection with Salmonella enterica serovar Typhi. We identified 7 in vivo induced (IVI) antigens, which included BcfD (a fimbrial structural subunit), GrxC (a glutaredoxin 3), SapB (an ABC-type transport system), T3663 (an ABC-type uncharacterized transport system), T3816 (a putative rhodanese-related sulfurtransferase), T1497 (a probable TonB-dependent receptor) and T3689 (unknown function). Of the 7 identified antigens, 5 antigens had no cross-immunoreactivity in adsorbed control sera from healthy subjects. These 5 included BcfD, GrxC, SapB, T3663 and T3689. Antigens identified in this study are potential targets for drug and vaccine development and may be utilized as diagnostic agents.展开更多
Persistent activation of Survivin and its overexpression contribute to the formation, progression and metastasis of several different tumor types. Therefore, Survivin is an ideal target for RNA interference mediated-g...Persistent activation of Survivin and its overexpression contribute to the formation, progression and metastasis of several different tumor types. Therefore, Survivin is an ideal target for RNA interference mediated-growth inhibition. Blockade of Survivin using specific short hairpin RNAs （shRNA） can significantly reduce prostate tumor growth. RNA interference does not fully ablate target gene expression, owing to the idiosyncrasies associated with shRNAs and their targets. To enhance the therapeutic efficacy of Survivin-specific shRNA, we employed a combinatorial expression of Survivin-specific shRNA and gene associated with retinoid-interferon-induced mortality-19 （GRIM-19）. Then, the GRIM-19 coding sequences and Survivin-specific shRNAs were used to create a dual expression plasmid vector and were carried by an attenuated strain of Salmonella enteric serovar typhimurium （S. typhimurium） to treat prostate cancer in vitro and in vivo. We found that the co-expressed Survivin-specific shRNA and GRIM-19 synergistically and more effectively inhibited prostate tumor proliferation and survival, when compared with treatment with either single agent alone in vitroand in vivo. This study has provided a novel cancer gene therapeutic approach for prostate cancer.展开更多
pRST98 is a chimeric plasmid isolated from Salmonella enterica serovar Typhi(S.typhi)that mediates the functions of drug resistance and virulence.Previously,we reported that Salmonella plasmid virulence(spv)genes were...pRST98 is a chimeric plasmid isolated from Salmonella enterica serovar Typhi(S.typhi)that mediates the functions of drug resistance and virulence.Previously,we reported that Salmonella plasmid virulence(spv)genes were present in S.typhi.In our current study,we investigated whether plasmid pRST98 exhibits significant cytotoxicity in macrophages.pRST98 was transferred into the avirulent Salmonella enterica serovar Typhimurium(S.typhimurium)strain RIA to create the transconjugant pRST98/RIA.The standard S.typhimurium virulent strain SR-11,which carries a 100-kb virulence plasmid,was used as a positive control.The bacterial strains were incubated with a murine macrophage-like cell line(J774A.1)in vitro.Apoptosis of J774A.1 cells was examined by electron microscopy and flow cytometry after annexin-V/propidium iodide labeling,and the survival of Salmonella strains in J774A.1 cells was determined.Results showed that macrophages infected with strain pRST98/RIA displayed greater levels of apoptosis than those infected with RIA and that pRST98 may increase bacterial survival in macrophages.Further studies showed that the pRST98-induced death of macrophages was associated with the loss of mitochondrial membrane potential and that pRST98 may activate caspase-9 and then caspase-3.The research data indicate that the virulence of bacteria that contain the pRST98 plasmid is enhanced;the presence of this plasmid increases the survival of the bacterial pathogen and acts through the mitochondrial pathway to mediate macrophage apoptosis.展开更多
BACKGROUND Secondary hemophagocytic lymphohistiocytosis(sHLH)triggered by Salmonella enterica serovar Typhimurium is rare in pediatric patients.There is no consensus on how to treat S.typhimurium-triggered sHLH.CASE S...BACKGROUND Secondary hemophagocytic lymphohistiocytosis(sHLH)triggered by Salmonella enterica serovar Typhimurium is rare in pediatric patients.There is no consensus on how to treat S.typhimurium-triggered sHLH.CASE SUMMARY A 9-year-old boy with intermittent fever for 3 d presented to our hospital with positive results for S.typhimurium,human rhinovirus,and Mycoplasma pneumoniae infections.At the time of admission to our institution,the patient’s T helper 1/T helper 2 cytokine levels were 326 pg/mL for interleukin 6(IL-6),9.1 pg/mL for IL-10,and 246.7 pg/mL for interferon-gamma(IFN-γ),for which the ratio of IL-10 to IFN-γwas 0.04.In this study,the patient received meropenem,linezolid,and cefoperazone/sulbactam in combination with high-dose methylprednisolone therapy(10 mg/kg/d for 3 d)and antishock supportive treatment twice.After careful evaluation,this patient did not receive HLH chemotherapy and recovered well.CONCLUSION S.Typhimurium infection-triggered sHLH patient had a ratio of IL-10 to IFN-γ≤1.33,an IL-10 concentration≤10.0 pg/mL,and/or an IFN-γconcentration≤225 pg/mL at admission.Early antimicrobial and supportive treatment was sufficient,and the HLH-94/2004 protocol was not necessary under these conditions.展开更多
Background： Salmonella enterica serovar Enteritidis （SE）, as a major cause of foodborn illness, infects humans mainly through the egg. However, the symptom of laying hens usually is not typical and hard to diagnosi...Background： Salmonella enterica serovar Enteritidis （SE）, as a major cause of foodborn illness, infects humans mainly through the egg. However, the symptom of laying hens usually is not typical and hard to diagnosis. In the present study, it is studied that the influences of SE infection on layers＇ performance, egg quality and blood biochemical indicators. It will help us to improve the strategy to control SE infection in commercial layers. One hundred layers at 20 wk of age were divided into 2 groups, 60 hens for experiment and others for control. The experiment group was fed with the dosage of 108 CFU SE per hen. The specific PCR was used to detect the deposition of SE. On the 8 d after SE infection, 10 hens from the control group and 30 hens from the experimenta group were slaughtered to detect the SE colonization. The production performance, egg quality and blood biochemical indices were also analyzed. Results： The results showed that the colonization rate of SE was highest in caecum contents （55.17%） and lowest in vagina （17.24%）. For the eggs the detection rate of SE was highest on the eggshell （80.00%） and lowest in yolk （18.81%）. SE infection had no significant influence on production performance and egg qualities （P 〉 0.05）. The difference of laying rate between the experimental and control groups was less than 0.30%, and both were approximately equal to 82.00%. The blood analysis showed that the aspartic aminotransferase （AST） and alanine aminotransferase （ALT） of experimental group was significantly higher than those of control group （P 〈 0.05）. For experimental and control groups AST values were 236.22 U/I and 211.84 U/I respectively, and ALT values were 32.19 U/I and 24.55 U/I. All of coefficients were less than 20%. The colonization of SE in organs increases the enzyme activities of AST and ALT in blood. Conclusions： SE in feed could invade the oviduct and infect the forming eggs. It significantly increased the concentration of ALT and AST in blood. Ho展开更多
Objective:To detect leptospiral antibodies by microscopic agglutination test(MAT) in northeast of Iran.Methods:This study was conducted to evaluate prevalence of human leptospiral infections by MAT.using six current r...Objective:To detect leptospiral antibodies by microscopic agglutination test(MAT) in northeast of Iran.Methods:This study was conducted to evaluate prevalence of human leptospiral infections by MAT.using six current reference strains of Leptospira interrogans in north-east of Iran.A total of 285 serum samples were collected from three north-east provinces of Iran,from December,2009 to June,2010.Results:Antibodies were detected at least against one serovar of Leptospira interrogans in 45 sera(15.79%) among 283 samples at a dilution 1:100 or greater.Positive titers against more than one serovar were detected in 24 sera of the positive samples.Therefore, there were 75 positive reactions against different serovar of Leptospira interrogans.Positive titers were recorded against serovar icterohaemorrhagiae(31 samples),hardjo(26 samples), grippotyphosa(7 samples),pomona(5 samples),canicola(4 samples) and ballum(2.sample). Conclusions:In present study the most prevalent(Leptospira icterohaemorrhagiae) and the least prevalent(Leptospira ballum) serovar are different from previous studies.Maybe,species and prevalence of serovars change during the lime in one area and between regions.展开更多
Objective:To develop allenualed slrains of Salmonella enterica serorar Typhi(S.typhi) for the candidate vaccine by osmolar stress.Mothods:S.typhi SS3 and SS5 strains were isolated from asymptomatic typhoid carriers in...Objective:To develop allenualed slrains of Salmonella enterica serorar Typhi(S.typhi) for the candidate vaccine by osmolar stress.Mothods:S.typhi SS3 and SS5 strains were isolated from asymptomatic typhoid carriers in Mamakkal,Tamil Nadu.India.Both strains were grown in LB(Luria Bertani) medium supplemented with various concentration of NaCl(0.1- 0.7M) respectively.The effecl of osmolar stress was determined at molecular level by PCR using MCR 06 and MCR07 primers corresponding to ompR with chromosomal DNA of S.typhi SS3 and SS5 strains.Attenuation by osmolar stress results in deletion mutation of the.S.typhi slrains was determined by agglutination assays,precipitation method.SDS PAGE analysis and by animal models.Results:The 799 bp amplified ompR gene product from wild type S.typhi SS3 and SS5 illustrate the presence of virulent gene.Interestingly,there was only a 282 bp amplified product from S.typhi SS3 and SS5 grown in the presence of 0.5.0.6 and 0.7 M NaCl.This illustrates the occurrence of deletion mutation in ompR gene al high concentration of NaCl.Furthermore,both the wildtype and mutant S.typhi outer membrane SDS-PAGF.profile reveals the differences in the expression of ompF.ompC and ompA proteins.In mice,wild type and mutant strains lethal dose(LD_(50)) were determined.The mice died within 72 h when both the wild type strains were injected intraperitoneally with 3 log CFU-mL^(-1).When the mice were injected with the mutants in same dosage,no clinical symptoms were observed;whereas the serum antibodv litre was elicited within two weeks indicated that the mutants have the ability to induce protective humoral immune response.These results suggest that S.typhi SS3 and SS5 may bo used as good candidate strains for the development of live attenuated vaccine against salmonellosis.Conclusions:This study demonstrates that the S.typhi strains were allenualed and could be good vaccine candidates in future.展开更多
基金Supported by the National Natural Science Foundation of China (Grant No. 30500435)
文摘During infectious disease episodes, pathogens express distinct subsets of virulence factors which allow them to adapt to different environments. Hence, genes that are expressed or upregulated in vivo are implicated in pathogenesis. We used in vivo induced antigen technology (IVIAT) to identify antigens which are expressed during infection with Salmonella enterica serovar Typhi. We identified 7 in vivo induced (IVI) antigens, which included BcfD (a fimbrial structural subunit), GrxC (a glutaredoxin 3), SapB (an ABC-type transport system), T3663 (an ABC-type uncharacterized transport system), T3816 (a putative rhodanese-related sulfurtransferase), T1497 (a probable TonB-dependent receptor) and T3689 (unknown function). Of the 7 identified antigens, 5 antigens had no cross-immunoreactivity in adsorbed control sera from healthy subjects. These 5 included BcfD, GrxC, SapB, T3663 and T3689. Antigens identified in this study are potential targets for drug and vaccine development and may be utilized as diagnostic agents.
基金This study was supported by the National Natural Science Foundation of China （No. 30801354 and No. 3097079J）, Jilin Provincial Science and Technology Department （No. 20080154 and No.20110146） and the PhD Programs Foundation of Ministry of Education of China （No. 200801831077） （Ling Zhang）
文摘Persistent activation of Survivin and its overexpression contribute to the formation, progression and metastasis of several different tumor types. Therefore, Survivin is an ideal target for RNA interference mediated-growth inhibition. Blockade of Survivin using specific short hairpin RNAs （shRNA） can significantly reduce prostate tumor growth. RNA interference does not fully ablate target gene expression, owing to the idiosyncrasies associated with shRNAs and their targets. To enhance the therapeutic efficacy of Survivin-specific shRNA, we employed a combinatorial expression of Survivin-specific shRNA and gene associated with retinoid-interferon-induced mortality-19 （GRIM-19）. Then, the GRIM-19 coding sequences and Survivin-specific shRNAs were used to create a dual expression plasmid vector and were carried by an attenuated strain of Salmonella enteric serovar typhimurium （S. typhimurium） to treat prostate cancer in vitro and in vivo. We found that the co-expressed Survivin-specific shRNA and GRIM-19 synergistically and more effectively inhibited prostate tumor proliferation and survival, when compared with treatment with either single agent alone in vitroand in vivo. This study has provided a novel cancer gene therapeutic approach for prostate cancer.
基金supported by the Natural Science Foundation of China(No.30972768)the Special and General Postdoctoral Science Foundation of China(No.200902529 and No.20080430178)+1 种基金the Natural Science Foundation of Jiangsu High Education Institute of China(No.08KJB310009)the Social Development Science Foundation of Suzhou City of China(No.SS08025).
文摘pRST98 is a chimeric plasmid isolated from Salmonella enterica serovar Typhi(S.typhi)that mediates the functions of drug resistance and virulence.Previously,we reported that Salmonella plasmid virulence(spv)genes were present in S.typhi.In our current study,we investigated whether plasmid pRST98 exhibits significant cytotoxicity in macrophages.pRST98 was transferred into the avirulent Salmonella enterica serovar Typhimurium(S.typhimurium)strain RIA to create the transconjugant pRST98/RIA.The standard S.typhimurium virulent strain SR-11,which carries a 100-kb virulence plasmid,was used as a positive control.The bacterial strains were incubated with a murine macrophage-like cell line(J774A.1)in vitro.Apoptosis of J774A.1 cells was examined by electron microscopy and flow cytometry after annexin-V/propidium iodide labeling,and the survival of Salmonella strains in J774A.1 cells was determined.Results showed that macrophages infected with strain pRST98/RIA displayed greater levels of apoptosis than those infected with RIA and that pRST98 may increase bacterial survival in macrophages.Further studies showed that the pRST98-induced death of macrophages was associated with the loss of mitochondrial membrane potential and that pRST98 may activate caspase-9 and then caspase-3.The research data indicate that the virulence of bacteria that contain the pRST98 plasmid is enhanced;the presence of this plasmid increases the survival of the bacterial pathogen and acts through the mitochondrial pathway to mediate macrophage apoptosis.
基金Supported by Zhejiang Province Health and Wellness Science and Technology Program in 2022,China,No.2022RC202.
文摘BACKGROUND Secondary hemophagocytic lymphohistiocytosis(sHLH)triggered by Salmonella enterica serovar Typhimurium is rare in pediatric patients.There is no consensus on how to treat S.typhimurium-triggered sHLH.CASE SUMMARY A 9-year-old boy with intermittent fever for 3 d presented to our hospital with positive results for S.typhimurium,human rhinovirus,and Mycoplasma pneumoniae infections.At the time of admission to our institution,the patient’s T helper 1/T helper 2 cytokine levels were 326 pg/mL for interleukin 6(IL-6),9.1 pg/mL for IL-10,and 246.7 pg/mL for interferon-gamma(IFN-γ),for which the ratio of IL-10 to IFN-γwas 0.04.In this study,the patient received meropenem,linezolid,and cefoperazone/sulbactam in combination with high-dose methylprednisolone therapy(10 mg/kg/d for 3 d)and antishock supportive treatment twice.After careful evaluation,this patient did not receive HLH chemotherapy and recovered well.CONCLUSION S.Typhimurium infection-triggered sHLH patient had a ratio of IL-10 to IFN-γ≤1.33,an IL-10 concentration≤10.0 pg/mL,and/or an IFN-γconcentration≤225 pg/mL at admission.Early antimicrobial and supportive treatment was sufficient,and the HLH-94/2004 protocol was not necessary under these conditions.
基金supported by National System for Layer Production Technology of China(CARS-41)Program for Changjiang Scholars and Innovative Research Team in University of Ministry of Education of China "Study on Poultry Eggs and Meat Detection Technology"(IRT0945)Chinese Universities Scientific Fund(2013QJ069)
文摘Background： Salmonella enterica serovar Enteritidis （SE）, as a major cause of foodborn illness, infects humans mainly through the egg. However, the symptom of laying hens usually is not typical and hard to diagnosis. In the present study, it is studied that the influences of SE infection on layers＇ performance, egg quality and blood biochemical indicators. It will help us to improve the strategy to control SE infection in commercial layers. One hundred layers at 20 wk of age were divided into 2 groups, 60 hens for experiment and others for control. The experiment group was fed with the dosage of 108 CFU SE per hen. The specific PCR was used to detect the deposition of SE. On the 8 d after SE infection, 10 hens from the control group and 30 hens from the experimenta group were slaughtered to detect the SE colonization. The production performance, egg quality and blood biochemical indices were also analyzed. Results： The results showed that the colonization rate of SE was highest in caecum contents （55.17%） and lowest in vagina （17.24%）. For the eggs the detection rate of SE was highest on the eggshell （80.00%） and lowest in yolk （18.81%）. SE infection had no significant influence on production performance and egg qualities （P 〉 0.05）. The difference of laying rate between the experimental and control groups was less than 0.30%, and both were approximately equal to 82.00%. The blood analysis showed that the aspartic aminotransferase （AST） and alanine aminotransferase （ALT） of experimental group was significantly higher than those of control group （P 〈 0.05）. For experimental and control groups AST values were 236.22 U/I and 211.84 U/I respectively, and ALT values were 32.19 U/I and 24.55 U/I. All of coefficients were less than 20%. The colonization of SE in organs increases the enzyme activities of AST and ALT in blood. Conclusions： SE in feed could invade the oviduct and infect the forming eggs. It significantly increased the concentration of ALT and AST in blood. Ho
基金Supported by a grant form Research Council of Shahid BahonarUniversity of Kerman.Iran
文摘Objective:To detect leptospiral antibodies by microscopic agglutination test(MAT) in northeast of Iran.Methods:This study was conducted to evaluate prevalence of human leptospiral infections by MAT.using six current reference strains of Leptospira interrogans in north-east of Iran.A total of 285 serum samples were collected from three north-east provinces of Iran,from December,2009 to June,2010.Results:Antibodies were detected at least against one serovar of Leptospira interrogans in 45 sera(15.79%) among 283 samples at a dilution 1:100 or greater.Positive titers against more than one serovar were detected in 24 sera of the positive samples.Therefore, there were 75 positive reactions against different serovar of Leptospira interrogans.Positive titers were recorded against serovar icterohaemorrhagiae(31 samples),hardjo(26 samples), grippotyphosa(7 samples),pomona(5 samples),canicola(4 samples) and ballum(2.sample). Conclusions:In present study the most prevalent(Leptospira icterohaemorrhagiae) and the least prevalent(Leptospira ballum) serovar are different from previous studies.Maybe,species and prevalence of serovars change during the lime in one area and between regions.
文摘Objective:To develop allenualed slrains of Salmonella enterica serorar Typhi(S.typhi) for the candidate vaccine by osmolar stress.Mothods:S.typhi SS3 and SS5 strains were isolated from asymptomatic typhoid carriers in Mamakkal,Tamil Nadu.India.Both strains were grown in LB(Luria Bertani) medium supplemented with various concentration of NaCl(0.1- 0.7M) respectively.The effecl of osmolar stress was determined at molecular level by PCR using MCR 06 and MCR07 primers corresponding to ompR with chromosomal DNA of S.typhi SS3 and SS5 strains.Attenuation by osmolar stress results in deletion mutation of the.S.typhi slrains was determined by agglutination assays,precipitation method.SDS PAGE analysis and by animal models.Results:The 799 bp amplified ompR gene product from wild type S.typhi SS3 and SS5 illustrate the presence of virulent gene.Interestingly,there was only a 282 bp amplified product from S.typhi SS3 and SS5 grown in the presence of 0.5.0.6 and 0.7 M NaCl.This illustrates the occurrence of deletion mutation in ompR gene al high concentration of NaCl.Furthermore,both the wildtype and mutant S.typhi outer membrane SDS-PAGF.profile reveals the differences in the expression of ompF.ompC and ompA proteins.In mice,wild type and mutant strains lethal dose(LD_(50)) were determined.The mice died within 72 h when both the wild type strains were injected intraperitoneally with 3 log CFU-mL^(-1).When the mice were injected with the mutants in same dosage,no clinical symptoms were observed;whereas the serum antibodv litre was elicited within two weeks indicated that the mutants have the ability to induce protective humoral immune response.These results suggest that S.typhi SS3 and SS5 may bo used as good candidate strains for the development of live attenuated vaccine against salmonellosis.Conclusions:This study demonstrates that the S.typhi strains were allenualed and could be good vaccine candidates in future.
