Objective This study investigated how the natural phytophenol and potent SIRT1 activator resveratrol(RSV)regulate necroptosis during Vibrio vulnificus(V.vulnificus)-induced sepsis and the potential mechanism.Methods T...Objective This study investigated how the natural phytophenol and potent SIRT1 activator resveratrol(RSV)regulate necroptosis during Vibrio vulnificus(V.vulnificus)-induced sepsis and the potential mechanism.Methods The effect of RSV on V.vulnificus cytolysin(VVC)-induced necroptosis was analyzed in vitro using CCK-8 and Western blot assays.Enzyme-linked immunosorbent assays and quantitative real-time polymerase chain reaction,western blot,and immunohistochemistry and survival analyses were performed to elucidate the effect and mechanism of RSV on necroptosis in a V.vulnificus-induced sepsis mouse model.Results RSV relieved necroptosis induced by VVC in RAW264.7 and MLE12 cells.RSV also inhibited the inflammatory response,had a protective effect on histopathological changes,and reduced the expression level of the necroptosis indicator pMLKL in peritoneal macrophages,lung,spleen,and liver tissues of V.vulnificus-induced septic mice in vivo.Pretreatment with RSV downregulated the mRNA of the necroptosis indicator and protein expression in peritoneal macrophages and tissues of V.vulnificusinduced septic mice.RSV also improved the survival of V.vulnificus-induced septic mice.Conclusion Our findings collectively demonstrate that RSV prevented V.vulnificus-induced sepsis by attenuating necroptosis,highlighting its potency in the clinical management of V.vulnificus-induced sepsis.展开更多
Vibrio vulnificus can cause rotten-skin disease in marine fish aquaculture, resulting in a large number of deaths. The occurrence of rotten-skin disease is becoming severer and severer, leading to serious economic los...Vibrio vulnificus can cause rotten-skin disease in marine fish aquaculture, resulting in a large number of deaths. The occurrence of rotten-skin disease is becoming severer and severer, leading to serious economic losses. In this study, the sensitivities of V. vulnificus to the 18 kinds of herbs, such as stone calamus and Chinese rhubarb, were determined using the agar diffusion method （susceptibility paper）. The results showed that Chinese gall, dark plum, sea buckthorn and eucalyptus leaves have bacteriostatic and bactericidal effects, while sappan wood, Chi- nese rhubarb, pomegranate bark, mentha and clove have strong bacteriostatic ef- fects. The Chinese medicinal herb bath showed good control effect on V. vulnificus.展开更多
Dynamic changes in mRNA expressions of liver tissue apoptosis-promoting genes Fas and Bax and apoptosis-inhibiting gene Bcl-2 of vibrio vulnificus sepsis rats were detected and the effects of antibacterial agents were...Dynamic changes in mRNA expressions of liver tissue apoptosis-promoting genes Fas and Bax and apoptosis-inhibiting gene Bcl-2 of vibrio vulnificus sepsis rats were detected and the effects of antibacterial agents were examined. The rat model with Vibrio vulnificus sepsis （VV group） was established and some of the Vibrio vulnificus sepsis rats were treated with antibacterial agents （AA group）. The mRNA expressions of Fas, Bax and Bcl-2 were measured by reverse transcription polymerase chain reaction （RT-PCR）. As compared with normal control group （NC group）, the ex- pressions of Fas and Bax mRNA in liver tissue at all different time points in VV group were increased significantly （P〈0.05）, and the highest levels of Fas and Bax mRNA expressions were 6 and 12 h after the infection, respectively. At the same time, the expression of Bcl-2 mRNA in liver tissue at all different time points in VV group were decreased significantly （P〈0.05）, and the lowest level of Bcl-2 mRNA expression appeared 2 h after the infection. The mRNA expressions of Bcl-2 in liver tissue 9 and 12 h after the infection in AA group were increased significantly （P〈0.05） compared with NC group, while the expressions of Fas and Bax mRNA were not significantly different from those of NC group. Compared with VV group, the expression of Fas mRNA in AA group was decreased （P〈0.05） and Bax rnRNA was decreased significantly 12 and 16 h after the infection （P〈0.05）, while the expressions of Bcl-2 mRNA were increased significantly 9, 12 and 16 h after the infection （P〈0.05）. It is concluded that the mRNA expressions of liver tissue apoptosis-promoting genes Fas and Bax were increased remarkably in vibrio vulnificus sepsis rats, whereas the expression of apoptosis-inhibiting gene Bcl-2 mRNA was decreased obviously in sepsis rats in early stage. The treatment with cefoperazone sodium and levofloxacin lactate could inhibit the expression of Fas mRNA and Bax mRNA and enhance the expression ofBcl-2 rnRNA at the same time.展开更多
Objective To establish a TaqMan real-time fluorescent quantitative PCR to detect Vibrio vulnificus based on the hemolysin gene （vvhA） coding cytolysin. Methods Primers and probes in the conserved region of the vvhA ...Objective To establish a TaqMan real-time fluorescent quantitative PCR to detect Vibrio vulnificus based on the hemolysin gene （vvhA） coding cytolysin. Methods Primers and probes in the conserved region of the vvhA gene sequence were designed for the TaqMan real-time PCR to detect 100 bp amplicon from V. vulnificus DNA. Recombinant plasmid pMD19-vvhA100 was constructed and used as a positive control during the detection. Minimal amplification cycles （Ct value） and fluorescence intensity enhancement （ARn value） were used as observing indexes to optimize the reaction conditions of TaqMan real-time PCR. The TaqMan assay for the detection of Vbirio vulnificus was evaluated in pure culture, mice tissue which artificially contaminated Vibrio vulnificus and clinical samples. Results The established TaqMan real-time PCR showed positive results only for Vibrio vulnificus DNA and pMD19-vvhA100. The standard curve was plotted and the minimum level of the vvhA target from the recombinant plasmid DNA was 103 copies with a Ct value of 37.94±0.19, as the equivalent of 0.01 ng purified genomic DNA of Vibrio vulnificus. The results detected by TaqMan PCR were positive for the 16 clinical samples and all the specimens of peripheral blood and subcutaneous tissue of mice which were infected with Vibrio vulnificus. Conclusion TaqMan real-time PCR is a rapid, effective, and quantitative tool to detect Vibro vulnificus, and can be used in clinical laboratory diagnosis of septicemia and wound infection caused by Vibrio vulnificus.展开更多
Vibrio vulnificus is a deadly disease that has been increasing in prevalence. In this study, we review both primary and secondary data to discuss the factors that are contributing to the increase of vibrio disease, ca...Vibrio vulnificus is a deadly disease that has been increasing in prevalence. In this study, we review both primary and secondary data to discuss the factors that are contributing to the increase of vibrio disease, causing a 41% increase between 1996 and 2005. It has also been shown that public health campaigns to limit vibrio infections have focused on raw seafood consumption. However, an estimated 42% of infections are now being caused by wound infection, rather than through contaminated seafood. This shows the disparities in addressing vibrio contamination through contaminated seawater and open wounds. This is particularly stressing as global warming is causing an increase in risk of contaminated seawater. Reasons for this increase are discussed, and also possible solutions are presented for public health interventions to help mitigate this rise in vibrio infections.展开更多
Objective: To summarize and analyze the clinical treatment experience of Vibrio vulnificus sepsis and improve the success rate of treatment. Methods: The clinical data of a patient with vibrio vulnificus sepsis admitt...Objective: To summarize and analyze the clinical treatment experience of Vibrio vulnificus sepsis and improve the success rate of treatment. Methods: The clinical data of a patient with vibrio vulnificus sepsis admitted to the Department of Critical Care Medicine, the First Affiliated Hospital of Hainan Medical College in October 2019 were analyzed. Summarize the experience of treatment and compare with other literatures to analyze the most reasonable clinical treatment of vibrio vulnificus sepsis. Results: The patient was acutely ill, the disease progressed rapidly, fever, soft tissue swelling and pain, necrosis, ulceration and pus, systemic multiple organ failure, family members finally gave up treatment, microbiology culture prompted: Vibrio parahaemolyticus. Conclusions: Early intervention, blood purification, debridement, removal of infection sources and inflammatory mediators can improve the survival rate of patients with Vibrio vulnificus infection. Due to the high mortality rate, clinicians should pay attention to it.展开更多
Purpose:Vibrio vulnificus (V. Vulnificus) infection is characterized by rapid onset, aggressive progression, and challenging treatment. Bacterial resistance poses a significant challenge for clinical anti-infection tr...Purpose:Vibrio vulnificus (V. Vulnificus) infection is characterized by rapid onset, aggressive progression, and challenging treatment. Bacterial resistance poses a significant challenge for clinical anti-infection treatment and is thus the subject of research. Enhancing host infection tolerance represents a novel infection prevention strategy to improve patient survival. Our team initially identified cytochrome P4501A1 (CYP1A1) as an important target owing to its negative modulation of the body''s infection tolerance. This study explored the superior effects of the CYP1A1 inhibitor bergamottin compared to antibiotic combination therapy on the survival of mice infected with multidrug-resistantV. Vulnificus and the protection of their vital organs.Methods:An increasing concentration gradient method was used to induce multidrug-resistantV. Vulnificus development. We established a lethal infection model in C57BL/6J male mice and evaluated the effect of bergamottin on mouse survival. A mild infection model was established in C57BL/6J male mice, and the serum levels of creatinine, urea nitrogen, aspartate aminotransferase, and alanine aminotransferase were determined using enzyme-linked immunosorbent assay to evaluate the effect of bergamottin on liver and kidney function. The morphological changes induced in the presence of bergamottin in mouse organs were evaluated by hematoxylin and eosin staining of liver and kidney tissues. The bacterial growth curve and organ load determination were used to evaluate whether bergamottin has a direct antibacterial effect on multidrug-resistantV. Vulnificus. Quantification of inflammatory factors in serum by enzyme-linked immunosorbent assay and the expression levels of inflammatory factors in liver and kidney tissues by real-time quantitative polymerase chain reaction were performed to evaluate the effect of bergamottin on inflammatory factor levels. Western blot analysis of IκBα, phosphorylated IκBα, p65, and phosphorylated p65 protein expression in liver and kidney tissues and in human hepatocellular carcinomas-2 and human kidney-2 cell lines was used to evaluate the effect of bergamottin on the nuclear factor kappa-B signaling pathway. One-way ANOVA and Kaplan-Meier analysis were used for statistical analysis.Results:In mice infected with multidrug-resistantV. Vulnificus, bergamottin prolonged survival (p = 0.014), reduced the serum creatinine (p = 0.002), urea nitrogen (p = 0.030), aspartate aminotransferase (p = 0.029), and alanine aminotransferase (p = 0.003) levels, and protected the cellular morphology of liver and kidney tissues. Bergamottin inhibited interleukin (IL)-1β, IL-6, and tumor necrosis factor (TNF)-α expression in serum (IL-1β:p = 0.010, IL-6:p = 0.029, TNF-α:p = 0.025) and inhibited the protein expression of the inflammatory factors IL-1β, IL-6, TNF-α in liver (IL-1β:p = 0.010, IL-6:p = 0.011, TNF-α:p = 0.037) and kidney (IL-1β:p = 0.016, IL-6:p = 0.011, TNF-α:p = 0.008) tissues. Bergamottin did not affect the proliferation of multidrug-resistantV. Vulnificus or the bacterial load in the mouse peritoneal lavage fluid (p = 0.225), liver (p = 0.186), or kidney (p = 0.637).Conclusion:Bergamottin enhances the tolerance of mice to multidrug-resistantV. Vulnificus infection. This study can serve as a reference and guide the development of novel clinical treatment strategies forV. Vulnificus.展开更多
Half-smooth tongue sole(Cynoglossus semilaevis)is regarded as a significant commercial marine fish species in China,and frequent outbreaks of vibriosis has led to substantial economic losses.In this study,we investiga...Half-smooth tongue sole(Cynoglossus semilaevis)is regarded as a significant commercial marine fish species in China,and frequent outbreaks of vibriosis has led to substantial economic losses.In this study,we investigated the molecular mechanisms underlying the involvement of the extracellular matrix(ECM)signaling pathway in the skin immune response of C.semilaevis infected with Vibrio vulnificus.The results showed that most differentially expressed proteins(DEPs)were identified through iTRAQ proteome analysis,and ten ECM-related proteins were screened at 12 and 36 h post-infection(hpi).Notably,several DEPs associated with ECM including HSPG,FN,COCH,LAMB1,LAMC1,TSPN,COL6A1,AMBP,HX and ITGA6 were tested for their expression at the transcriptional level during V.vulnificus infection using qRT-PCR assay.The analysis of protein-protein interaction(PPI)showed that the integrin ITGA6 exhibited obvious interactions among ECM-related DEPs.Furthermore,the spatio-temporal expression of the Csitga6 gene was highest in the skin,gill,muscle,and spleen,but lower in the liver,kidney and intestine.To our knowledge,this is the first report on the involvement of ECM pathway in the skin immune response to V.vulnificus infection,and provides a reference for further study of the ECM mechanism in the mucosal immune response of marine fish.展开更多
Background Septicemia and inflammation-mediated septic shock caused by Vibrio vulnificus （VV） is strongly associated with chronic liver disease. This study examined the effects of antimicrobial therapy on expression...Background Septicemia and inflammation-mediated septic shock caused by Vibrio vulnificus （VV） is strongly associated with chronic liver disease. This study examined the effects of antimicrobial therapy on expression of hepatic toll-like receptors and inflammatory cytokines in rats with alcohol-induced liver disease complicated by VV sepsis. Methods Male Sprague-Dawley rats were assigned to the following treatment groups： normal control （N）, alcoholic liver disease control （A）, antimicrobial-treated alcoholic liver disease control （AA）, alcoholic liver disease with VV sepsis （AV）, and antimicrobial-treated alcoholic liver disease with VV sepsis （AVA）. Alcohol-induced liver disease was observed in all groups except N. Expression of mRNAs encoding hepatic toll-like receptors 2 and 4, myeloid differentiation protein-2, tumor necrosis factor-α （TNF-α）, interleukin （IL）-1β, IL-6 and IL-10 was determined by RT-PCR. Results mRNAs encoding toll-like receptors 2 and 4 and myeloid differentiation protein-2 were significantly up-regulated in group AV as compared to control groups at 2-24 hours of sepsis; peak expression occurred at 12 hours. These mRNAs were also up-regulated in group AVA but to lesser degrees than in group AV at comparable time post-infection, mRNAs encoding TNF-α, IL-1β and IL-6 were significantly elevated in group AV as a function of infection. In group AVA as compared to AV, expression of TNF-α and IL-1β mRNAs was lower at 12-24 hours post-infection and expression of IL-6 mRNA was lower at 24 hours post-infection. Compared with control groups, IL-10 mRNA expression in group AV was markedly higher at 12-24 hours of sepsis. Expression of IL-10 mRNA was lower in group AVA as compared to AV at 24 hours of sepsis. Conclusions Antimicrobial therapy reduces expression of toll-like receptors and cytokines in rats with alcohol-induced liver disease complicated by VV sepsis. Monitoring hepatic toll-like receptor and cytokine expression during antibiotic therapy may be valuable for determining the course of VV sepsis in subjects with liver disease.展开更多
Necrotizing fasciitis （NF） caused by vibrio vulnificus （VV） is a rare but fatal clinical emergency characterized by hemorrhagic bullous skin lesions,septic shock and multiple organ dysfunction syndrome （MODS）,co...Necrotizing fasciitis （NF） caused by vibrio vulnificus （VV） is a rare but fatal clinical emergency characterized by hemorrhagic bullous skin lesions,septic shock and multiple organ dysfunction syndrome （MODS）,commonly occurring in patients with chronic liver disease.1,2 In general,mortality of these patients is higher than 60%,which could approach 100% when complicated with septic shock.Therefore,early diagnosis and effectively surgical intervention in emergency room are the mainstays for better outcomes of these patients.However,many emergency physicians lack experiences in diagnosing and managing vibrio NF.Furthermore,due to the preexisting severe medical complications including hypotension,lactic acidosis,coagulation disorders and thrombocytopenia,the optimal therapeutic strategies for these critical patients should obviously be evaluated on a patient-to-patient basis,which remains largely undefined.展开更多
Background Vibrio vulnificus （Vv） is an estuadne bacterium that can cause primary septicemia as well as serious wound infections. However, little is known about the mechanisms by which Vv infects dendritic cells （...Background Vibrio vulnificus （Vv） is an estuadne bacterium that can cause primary septicemia as well as serious wound infections. However, little is known about the mechanisms by which Vv infects dendritic cells （DCs） and its effects on cytoskeleton. In this study, we aimed to investigate the invasion, internalization, and the organelles damage of the cultured dendritic cells （a DC 2.4 strain） during Vv infection. Methods The study model was the cultured DCs infected by a Vv 1.758 strain. Electron microscopy was used to observe the localization of bacteria at the different time points of infection, cell morphology, and the process of organelles changes. The cytoskeleton structure including the microfilaments and the microtubules rearrangement was examined under a fluorescence microscope. Results The Vv were pinocytosised into the DC cells through double-sides, and localized at 1-2 μm of the inner side membrane. It took 1.3, 1.9, and 3.4 hours to reach the infection ratio of 25%, 50%, and 75%, respectively. Using electron microscopy, the DCs had been observed to have developed chromatin aggregation within 4.0 hours, and significant cytoskeleton structure disruption was noted within 6.0 hours.展开更多
Vibrio vulnificus is an opportunistic pathogen widely distributed in estuarine and coastal seawaters. In this study, a culture-free method was developed to rapid detection of Vo vulnificus in all seasons, based on loo...Vibrio vulnificus is an opportunistic pathogen widely distributed in estuarine and coastal seawaters. In this study, a culture-free method was developed to rapid detection of Vo vulnificus in all seasons, based on loop-mediated isothermal amplification （LAMP） targeting virulent-correlated gene （vcg）. The new assay method allows differentiation between the virulent and non-virulent strain of V. vulnificus accurately. This method also allows effective detection of the pathogeny in winter when the bacterium lives in the viable but non-culturable （VBNC） state. A total of 30 costal seawater samples collected in all seasons were used for the evaluation of this method. The results show that the method is sensitive, accurate and convenient.展开更多
BACKGROUND We report on a case of Vibrio vulnificus(V.vulnificus)detected by metagenomics next-generation sequencing(mNGS)in a 53-year-old male patient with polymicrobial gas gangrene and successful treatment by surge...BACKGROUND We report on a case of Vibrio vulnificus(V.vulnificus)detected by metagenomics next-generation sequencing(mNGS)in a 53-year-old male patient with polymicrobial gas gangrene and successful treatment by surgery.This report raises awareness among dermatologists that when a patient is clinically suspected of a special type of pathogenic infection,the mNGS method should be preferred to identify the patient’s pathogen infection as soon as possible and then take effective treatment in time to save patients’lives.CASE SUMMARY A 53-year-old male who worked in the aquatic market complained of redness and swelling of the lower limbs,blisters and ulcers with fever for 3 d.We used mNGS to test the pathogens in ulcer secretions.The results were returned in 24 h and indicated:V.vulnificus,Fusobacterium necrophorum,Staphylococcus haemolyticus,Staphylococcus aureus,Streptococcus dysgalactiae and Klebsiella aerogenes.This patient was diagnosed with V.vulnificus infection.The emergency operation was performed immediately under combined lumbar and epidural anesthesia:Left leg expansion and exploration(August 10,2021).After surgery,we continued to use piperacillin sodium tazobactam sodium 4.5 g every 8 h and levofloxacin 0.5 g for anti-infection treatment.The patient underwent further surgery under lumbar anesthesia on August 17,2021 and August 31,2021:Left leg deactivation and skin grafting,negative pressure closed drainage and right thigh skin removal.After treatment,the transplanted flap survived.CONCLUSION We could confirm the diagnosis of Vibrio vulnificus infection within 24 h through mNGS detection and then immediately performed emergency surgery.展开更多
Objective To investigate the occurrence of important foodbome pathogens in shellstock Pacific oysters in the food markets in South China. Methods From July 2007 to June 2008, retail oysters were collected in different...Objective To investigate the occurrence of important foodbome pathogens in shellstock Pacific oysters in the food markets in South China. Methods From July 2007 to June 2008, retail oysters were collected in different seasons from South China and analyzed for the prevalence and levels of Listeria monocytogenes, Vibrio vulnificus and Vibrio parahaemolyticus. Results None of L. monocytogenes could be detected in any of the 202 oyster samples tested, while E vulnificus and E parahaemolyticus could be detected in 67 （54.9%） and 109 （89.3%） of the 122 oyster samples analyzed, respectively, with an MPN （most probable number） value greater than or equal to 3. V. wdnificus and Eparahaemolyticus with a more than 102 MPN/g were found in 36 （29.5%） and 59 （48.4%） of the 122 oyster samples, respectively. The tdh and trh genes were detected in 4 （0.3%） and 8 （0.6%） of the 1 349 V. parahaemolyticus isolates, respectively. Of the 122 samples, 4 （3.3%） was positive for either tdh or trh. The levels of E vulnificus and total V. parahaemolyticus in oysters in South China varied in different seasons. Conclusion V. vulnificus and pathogenic V. parahaemolyticus are frequently found in oysters in south China, which may pose a potential threat to public health. Data presented here will be useful for the microbiological risk assessment in oysters in China.展开更多
Wound infection is becoming a considerable healthcare crisis due to the abuse of antibiotics and the substantial production of multidrug-resistant bacteria.Seawater immersion wounds usually become a mortal trouble bec...Wound infection is becoming a considerable healthcare crisis due to the abuse of antibiotics and the substantial production of multidrug-resistant bacteria.Seawater immersion wounds usually become a mortal trouble because of the infection of Vibrio vulnificus.Bdellovibrio bacteriovorus,one kind of natural predatory bacteria,is recognized as a promising biological therapy against intractable bacteria.Here,we prepared a B.bacteriovorus-loaded polyvinyl alcohol/alginate hydrogel for the topical treatment of the seawater immersion wounds infected by V.vulnificus.The B.bacteriovorus-loaded hydrogel(BG)owned highly microporous structures with the mean pore size of 90μm,improving the rapid release of B.bacteriovorus from BG when contacting the aqueous surroundings.BG showed high biosafety with no L929 cell toxicity or hemolysis.More importantly,BG exhibited excellent in vitro anti-V.vulnificus effect.The highly effective infected wound treatment effect of BG was evaluated on mouse models,revealing significant reduction of local V.vulnificus,accelerated wound contraction,and alleviated inflammation.Besides the high bacterial inhibition of BG,BG remarkably reduced inflammatory response,promoted collagen deposition,neovascularization and re-epithelization,contributing to wound healing.BG is a promising topical biological formulation against infected wounds.展开更多
基金supported by the Beijing Municipal Natural Science Foundation[7204314]the Medical Innovation Project Foundation[CX19027]the Young Researcher Supporting Program[QNF19066]of the Chinese PLA General Hospital.
文摘Objective This study investigated how the natural phytophenol and potent SIRT1 activator resveratrol(RSV)regulate necroptosis during Vibrio vulnificus(V.vulnificus)-induced sepsis and the potential mechanism.Methods The effect of RSV on V.vulnificus cytolysin(VVC)-induced necroptosis was analyzed in vitro using CCK-8 and Western blot assays.Enzyme-linked immunosorbent assays and quantitative real-time polymerase chain reaction,western blot,and immunohistochemistry and survival analyses were performed to elucidate the effect and mechanism of RSV on necroptosis in a V.vulnificus-induced sepsis mouse model.Results RSV relieved necroptosis induced by VVC in RAW264.7 and MLE12 cells.RSV also inhibited the inflammatory response,had a protective effect on histopathological changes,and reduced the expression level of the necroptosis indicator pMLKL in peritoneal macrophages,lung,spleen,and liver tissues of V.vulnificus-induced septic mice in vivo.Pretreatment with RSV downregulated the mRNA of the necroptosis indicator and protein expression in peritoneal macrophages and tissues of V.vulnificusinduced septic mice.RSV also improved the survival of V.vulnificus-induced septic mice.Conclusion Our findings collectively demonstrate that RSV prevented V.vulnificus-induced sepsis by attenuating necroptosis,highlighting its potency in the clinical management of V.vulnificus-induced sepsis.
基金Supported by Incentive Program Subsidized by Department of Science and Technology of Hebei Province(15926620H)Key Technology Research and Development Program of Science and Technology Bureau of Qinhuangdao City(201401A067,201402B043)~~
文摘Vibrio vulnificus can cause rotten-skin disease in marine fish aquaculture, resulting in a large number of deaths. The occurrence of rotten-skin disease is becoming severer and severer, leading to serious economic losses. In this study, the sensitivities of V. vulnificus to the 18 kinds of herbs, such as stone calamus and Chinese rhubarb, were determined using the agar diffusion method （susceptibility paper）. The results showed that Chinese gall, dark plum, sea buckthorn and eucalyptus leaves have bacteriostatic and bactericidal effects, while sappan wood, Chi- nese rhubarb, pomegranate bark, mentha and clove have strong bacteriostatic ef- fects. The Chinese medicinal herb bath showed good control effect on V. vulnificus.
基金supported by a grant for supporting key research programs from Zhejiang Province (No.07-F04)a Science and Technology Fund of Zhejiang Province (No.2005C30009)
文摘Dynamic changes in mRNA expressions of liver tissue apoptosis-promoting genes Fas and Bax and apoptosis-inhibiting gene Bcl-2 of vibrio vulnificus sepsis rats were detected and the effects of antibacterial agents were examined. The rat model with Vibrio vulnificus sepsis （VV group） was established and some of the Vibrio vulnificus sepsis rats were treated with antibacterial agents （AA group）. The mRNA expressions of Fas, Bax and Bcl-2 were measured by reverse transcription polymerase chain reaction （RT-PCR）. As compared with normal control group （NC group）, the ex- pressions of Fas and Bax mRNA in liver tissue at all different time points in VV group were increased significantly （P〈0.05）, and the highest levels of Fas and Bax mRNA expressions were 6 and 12 h after the infection, respectively. At the same time, the expression of Bcl-2 mRNA in liver tissue at all different time points in VV group were decreased significantly （P〈0.05）, and the lowest level of Bcl-2 mRNA expression appeared 2 h after the infection. The mRNA expressions of Bcl-2 in liver tissue 9 and 12 h after the infection in AA group were increased significantly （P〈0.05） compared with NC group, while the expressions of Fas and Bax mRNA were not significantly different from those of NC group. Compared with VV group, the expression of Fas mRNA in AA group was decreased （P〈0.05） and Bax rnRNA was decreased significantly 12 and 16 h after the infection （P〈0.05）, while the expressions of Bcl-2 mRNA were increased significantly 9, 12 and 16 h after the infection （P〈0.05）. It is concluded that the mRNA expressions of liver tissue apoptosis-promoting genes Fas and Bax were increased remarkably in vibrio vulnificus sepsis rats, whereas the expression of apoptosis-inhibiting gene Bcl-2 mRNA was decreased obviously in sepsis rats in early stage. The treatment with cefoperazone sodium and levofloxacin lactate could inhibit the expression of Fas mRNA and Bax mRNA and enhance the expression ofBcl-2 rnRNA at the same time.
文摘Objective To establish a TaqMan real-time fluorescent quantitative PCR to detect Vibrio vulnificus based on the hemolysin gene （vvhA） coding cytolysin. Methods Primers and probes in the conserved region of the vvhA gene sequence were designed for the TaqMan real-time PCR to detect 100 bp amplicon from V. vulnificus DNA. Recombinant plasmid pMD19-vvhA100 was constructed and used as a positive control during the detection. Minimal amplification cycles （Ct value） and fluorescence intensity enhancement （ARn value） were used as observing indexes to optimize the reaction conditions of TaqMan real-time PCR. The TaqMan assay for the detection of Vbirio vulnificus was evaluated in pure culture, mice tissue which artificially contaminated Vibrio vulnificus and clinical samples. Results The established TaqMan real-time PCR showed positive results only for Vibrio vulnificus DNA and pMD19-vvhA100. The standard curve was plotted and the minimum level of the vvhA target from the recombinant plasmid DNA was 103 copies with a Ct value of 37.94±0.19, as the equivalent of 0.01 ng purified genomic DNA of Vibrio vulnificus. The results detected by TaqMan PCR were positive for the 16 clinical samples and all the specimens of peripheral blood and subcutaneous tissue of mice which were infected with Vibrio vulnificus. Conclusion TaqMan real-time PCR is a rapid, effective, and quantitative tool to detect Vibro vulnificus, and can be used in clinical laboratory diagnosis of septicemia and wound infection caused by Vibrio vulnificus.
文摘Vibrio vulnificus is a deadly disease that has been increasing in prevalence. In this study, we review both primary and secondary data to discuss the factors that are contributing to the increase of vibrio disease, causing a 41% increase between 1996 and 2005. It has also been shown that public health campaigns to limit vibrio infections have focused on raw seafood consumption. However, an estimated 42% of infections are now being caused by wound infection, rather than through contaminated seafood. This shows the disparities in addressing vibrio contamination through contaminated seawater and open wounds. This is particularly stressing as global warming is causing an increase in risk of contaminated seawater. Reasons for this increase are discussed, and also possible solutions are presented for public health interventions to help mitigate this rise in vibrio infections.
基金NSFC(81860347)Hainan natural science foundation(818MS140)+1 种基金Innovation program of Hainan association for science and technology for young talents in science and technology(QCXM201816)Hainan health and family planning industry research project(18A200178)
文摘Objective: To summarize and analyze the clinical treatment experience of Vibrio vulnificus sepsis and improve the success rate of treatment. Methods: The clinical data of a patient with vibrio vulnificus sepsis admitted to the Department of Critical Care Medicine, the First Affiliated Hospital of Hainan Medical College in October 2019 were analyzed. Summarize the experience of treatment and compare with other literatures to analyze the most reasonable clinical treatment of vibrio vulnificus sepsis. Results: The patient was acutely ill, the disease progressed rapidly, fever, soft tissue swelling and pain, necrosis, ulceration and pus, systemic multiple organ failure, family members finally gave up treatment, microbiology culture prompted: Vibrio parahaemolyticus. Conclusions: Early intervention, blood purification, debridement, removal of infection sources and inflammatory mediators can improve the survival rate of patients with Vibrio vulnificus infection. Due to the high mortality rate, clinicians should pay attention to it.
基金supported by the National Natural Science Foundation of China(grant number:82104247)Chongqing Talent Innovation Leading Plan(grant number:cstc2021ycjhbgzxm0340)+2 种基金Chongqing Doctor Express Project(grant number:CSTB2022BSXM-JCX0024)Hainan Clinical Medical Research Center Project(grant number:LCYX202205)Hainan Province Health Industry Research Project(grant number:22A200082).
文摘Purpose:Vibrio vulnificus (V. Vulnificus) infection is characterized by rapid onset, aggressive progression, and challenging treatment. Bacterial resistance poses a significant challenge for clinical anti-infection treatment and is thus the subject of research. Enhancing host infection tolerance represents a novel infection prevention strategy to improve patient survival. Our team initially identified cytochrome P4501A1 (CYP1A1) as an important target owing to its negative modulation of the body''s infection tolerance. This study explored the superior effects of the CYP1A1 inhibitor bergamottin compared to antibiotic combination therapy on the survival of mice infected with multidrug-resistantV. Vulnificus and the protection of their vital organs.Methods:An increasing concentration gradient method was used to induce multidrug-resistantV. Vulnificus development. We established a lethal infection model in C57BL/6J male mice and evaluated the effect of bergamottin on mouse survival. A mild infection model was established in C57BL/6J male mice, and the serum levels of creatinine, urea nitrogen, aspartate aminotransferase, and alanine aminotransferase were determined using enzyme-linked immunosorbent assay to evaluate the effect of bergamottin on liver and kidney function. The morphological changes induced in the presence of bergamottin in mouse organs were evaluated by hematoxylin and eosin staining of liver and kidney tissues. The bacterial growth curve and organ load determination were used to evaluate whether bergamottin has a direct antibacterial effect on multidrug-resistantV. Vulnificus. Quantification of inflammatory factors in serum by enzyme-linked immunosorbent assay and the expression levels of inflammatory factors in liver and kidney tissues by real-time quantitative polymerase chain reaction were performed to evaluate the effect of bergamottin on inflammatory factor levels. Western blot analysis of IκBα, phosphorylated IκBα, p65, and phosphorylated p65 protein expression in liver and kidney tissues and in human hepatocellular carcinomas-2 and human kidney-2 cell lines was used to evaluate the effect of bergamottin on the nuclear factor kappa-B signaling pathway. One-way ANOVA and Kaplan-Meier analysis were used for statistical analysis.Results:In mice infected with multidrug-resistantV. Vulnificus, bergamottin prolonged survival (p = 0.014), reduced the serum creatinine (p = 0.002), urea nitrogen (p = 0.030), aspartate aminotransferase (p = 0.029), and alanine aminotransferase (p = 0.003) levels, and protected the cellular morphology of liver and kidney tissues. Bergamottin inhibited interleukin (IL)-1β, IL-6, and tumor necrosis factor (TNF)-α expression in serum (IL-1β:p = 0.010, IL-6:p = 0.029, TNF-α:p = 0.025) and inhibited the protein expression of the inflammatory factors IL-1β, IL-6, TNF-α in liver (IL-1β:p = 0.010, IL-6:p = 0.011, TNF-α:p = 0.037) and kidney (IL-1β:p = 0.016, IL-6:p = 0.011, TNF-α:p = 0.008) tissues. Bergamottin did not affect the proliferation of multidrug-resistantV. Vulnificus or the bacterial load in the mouse peritoneal lavage fluid (p = 0.225), liver (p = 0.186), or kidney (p = 0.637).Conclusion:Bergamottin enhances the tolerance of mice to multidrug-resistantV. Vulnificus infection. This study can serve as a reference and guide the development of novel clinical treatment strategies forV. Vulnificus.
基金supported by the National Natural Science Foundation of China(no.32273182)the Natural Science Foundation of Tianjin City(no.23JCZDJC00240)+1 种基金Tianjin Agricultural Development Service Center Youth Science and Technology Innovation Project in 2024(no.zxkj202437)Teaching Program of Tianjin Agricultural University(2021-A-01).
文摘Half-smooth tongue sole(Cynoglossus semilaevis)is regarded as a significant commercial marine fish species in China,and frequent outbreaks of vibriosis has led to substantial economic losses.In this study,we investigated the molecular mechanisms underlying the involvement of the extracellular matrix(ECM)signaling pathway in the skin immune response of C.semilaevis infected with Vibrio vulnificus.The results showed that most differentially expressed proteins(DEPs)were identified through iTRAQ proteome analysis,and ten ECM-related proteins were screened at 12 and 36 h post-infection(hpi).Notably,several DEPs associated with ECM including HSPG,FN,COCH,LAMB1,LAMC1,TSPN,COL6A1,AMBP,HX and ITGA6 were tested for their expression at the transcriptional level during V.vulnificus infection using qRT-PCR assay.The analysis of protein-protein interaction(PPI)showed that the integrin ITGA6 exhibited obvious interactions among ECM-related DEPs.Furthermore,the spatio-temporal expression of the Csitga6 gene was highest in the skin,gill,muscle,and spleen,but lower in the liver,kidney and intestine.To our knowledge,this is the first report on the involvement of ECM pathway in the skin immune response to V.vulnificus infection,and provides a reference for further study of the ECM mechanism in the mucosal immune response of marine fish.
文摘Background Septicemia and inflammation-mediated septic shock caused by Vibrio vulnificus （VV） is strongly associated with chronic liver disease. This study examined the effects of antimicrobial therapy on expression of hepatic toll-like receptors and inflammatory cytokines in rats with alcohol-induced liver disease complicated by VV sepsis. Methods Male Sprague-Dawley rats were assigned to the following treatment groups： normal control （N）, alcoholic liver disease control （A）, antimicrobial-treated alcoholic liver disease control （AA）, alcoholic liver disease with VV sepsis （AV）, and antimicrobial-treated alcoholic liver disease with VV sepsis （AVA）. Alcohol-induced liver disease was observed in all groups except N. Expression of mRNAs encoding hepatic toll-like receptors 2 and 4, myeloid differentiation protein-2, tumor necrosis factor-α （TNF-α）, interleukin （IL）-1β, IL-6 and IL-10 was determined by RT-PCR. Results mRNAs encoding toll-like receptors 2 and 4 and myeloid differentiation protein-2 were significantly up-regulated in group AV as compared to control groups at 2-24 hours of sepsis; peak expression occurred at 12 hours. These mRNAs were also up-regulated in group AVA but to lesser degrees than in group AV at comparable time post-infection, mRNAs encoding TNF-α, IL-1β and IL-6 were significantly elevated in group AV as a function of infection. In group AVA as compared to AV, expression of TNF-α and IL-1β mRNAs was lower at 12-24 hours post-infection and expression of IL-6 mRNA was lower at 24 hours post-infection. Compared with control groups, IL-10 mRNA expression in group AV was markedly higher at 12-24 hours of sepsis. Expression of IL-10 mRNA was lower in group AVA as compared to AV at 24 hours of sepsis. Conclusions Antimicrobial therapy reduces expression of toll-like receptors and cytokines in rats with alcohol-induced liver disease complicated by VV sepsis. Monitoring hepatic toll-like receptor and cytokine expression during antibiotic therapy may be valuable for determining the course of VV sepsis in subjects with liver disease.
文摘Necrotizing fasciitis （NF） caused by vibrio vulnificus （VV） is a rare but fatal clinical emergency characterized by hemorrhagic bullous skin lesions,septic shock and multiple organ dysfunction syndrome （MODS）,commonly occurring in patients with chronic liver disease.1,2 In general,mortality of these patients is higher than 60%,which could approach 100% when complicated with septic shock.Therefore,early diagnosis and effectively surgical intervention in emergency room are the mainstays for better outcomes of these patients.However,many emergency physicians lack experiences in diagnosing and managing vibrio NF.Furthermore,due to the preexisting severe medical complications including hypotension,lactic acidosis,coagulation disorders and thrombocytopenia,the optimal therapeutic strategies for these critical patients should obviously be evaluated on a patient-to-patient basis,which remains largely undefined.
文摘Background Vibrio vulnificus （Vv） is an estuadne bacterium that can cause primary septicemia as well as serious wound infections. However, little is known about the mechanisms by which Vv infects dendritic cells （DCs） and its effects on cytoskeleton. In this study, we aimed to investigate the invasion, internalization, and the organelles damage of the cultured dendritic cells （a DC 2.4 strain） during Vv infection. Methods The study model was the cultured DCs infected by a Vv 1.758 strain. Electron microscopy was used to observe the localization of bacteria at the different time points of infection, cell morphology, and the process of organelles changes. The cytoskeleton structure including the microfilaments and the microtubules rearrangement was examined under a fluorescence microscope. Results The Vv were pinocytosised into the DC cells through double-sides, and localized at 1-2 μm of the inner side membrane. It took 1.3, 1.9, and 3.4 hours to reach the infection ratio of 25%, 50%, and 75%, respectively. Using electron microscopy, the DCs had been observed to have developed chromatin aggregation within 4.0 hours, and significant cytoskeleton structure disruption was noted within 6.0 hours.
基金The National High-Tech Research and Development Program of China ("863") under contract No.2006AA09Z170
文摘Vibrio vulnificus is an opportunistic pathogen widely distributed in estuarine and coastal seawaters. In this study, a culture-free method was developed to rapid detection of Vo vulnificus in all seasons, based on loop-mediated isothermal amplification （LAMP） targeting virulent-correlated gene （vcg）. The new assay method allows differentiation between the virulent and non-virulent strain of V. vulnificus accurately. This method also allows effective detection of the pathogeny in winter when the bacterium lives in the viable but non-culturable （VBNC） state. A total of 30 costal seawater samples collected in all seasons were used for the evaluation of this method. The results show that the method is sensitive, accurate and convenient.
基金National Natural Science Foundation of China,No.82173437and Natural Science Foundation of Guangdong Province,No.2020A15150875.
文摘BACKGROUND We report on a case of Vibrio vulnificus(V.vulnificus)detected by metagenomics next-generation sequencing(mNGS)in a 53-year-old male patient with polymicrobial gas gangrene and successful treatment by surgery.This report raises awareness among dermatologists that when a patient is clinically suspected of a special type of pathogenic infection,the mNGS method should be preferred to identify the patient’s pathogen infection as soon as possible and then take effective treatment in time to save patients’lives.CASE SUMMARY A 53-year-old male who worked in the aquatic market complained of redness and swelling of the lower limbs,blisters and ulcers with fever for 3 d.We used mNGS to test the pathogens in ulcer secretions.The results were returned in 24 h and indicated:V.vulnificus,Fusobacterium necrophorum,Staphylococcus haemolyticus,Staphylococcus aureus,Streptococcus dysgalactiae and Klebsiella aerogenes.This patient was diagnosed with V.vulnificus infection.The emergency operation was performed immediately under combined lumbar and epidural anesthesia:Left leg expansion and exploration(August 10,2021).After surgery,we continued to use piperacillin sodium tazobactam sodium 4.5 g every 8 h and levofloxacin 0.5 g for anti-infection treatment.The patient underwent further surgery under lumbar anesthesia on August 17,2021 and August 31,2021:Left leg deactivation and skin grafting,negative pressure closed drainage and right thigh skin removal.After treatment,the transplanted flap survived.CONCLUSION We could confirm the diagnosis of Vibrio vulnificus infection within 24 h through mNGS detection and then immediately performed emergency surgery.
基金supported by grants from the Key Projects in the National Science and Technology Pillar Program of China in the Eleventh Five-year Plan Period (Contract No. 2006BAK02A15)National Natural Science Foundation of China (Contract No. 30600499)
文摘Objective To investigate the occurrence of important foodbome pathogens in shellstock Pacific oysters in the food markets in South China. Methods From July 2007 to June 2008, retail oysters were collected in different seasons from South China and analyzed for the prevalence and levels of Listeria monocytogenes, Vibrio vulnificus and Vibrio parahaemolyticus. Results None of L. monocytogenes could be detected in any of the 202 oyster samples tested, while E vulnificus and E parahaemolyticus could be detected in 67 （54.9%） and 109 （89.3%） of the 122 oyster samples analyzed, respectively, with an MPN （most probable number） value greater than or equal to 3. V. wdnificus and Eparahaemolyticus with a more than 102 MPN/g were found in 36 （29.5%） and 59 （48.4%） of the 122 oyster samples, respectively. The tdh and trh genes were detected in 4 （0.3%） and 8 （0.6%） of the 1 349 V. parahaemolyticus isolates, respectively. Of the 122 samples, 4 （3.3%） was positive for either tdh or trh. The levels of E vulnificus and total V. parahaemolyticus in oysters in South China varied in different seasons. Conclusion V. vulnificus and pathogenic V. parahaemolyticus are frequently found in oysters in south China, which may pose a potential threat to public health. Data presented here will be useful for the microbiological risk assessment in oysters in China.
基金supported by the National Natural Science Foundation of China(82073791)。
文摘Wound infection is becoming a considerable healthcare crisis due to the abuse of antibiotics and the substantial production of multidrug-resistant bacteria.Seawater immersion wounds usually become a mortal trouble because of the infection of Vibrio vulnificus.Bdellovibrio bacteriovorus,one kind of natural predatory bacteria,is recognized as a promising biological therapy against intractable bacteria.Here,we prepared a B.bacteriovorus-loaded polyvinyl alcohol/alginate hydrogel for the topical treatment of the seawater immersion wounds infected by V.vulnificus.The B.bacteriovorus-loaded hydrogel(BG)owned highly microporous structures with the mean pore size of 90μm,improving the rapid release of B.bacteriovorus from BG when contacting the aqueous surroundings.BG showed high biosafety with no L929 cell toxicity or hemolysis.More importantly,BG exhibited excellent in vitro anti-V.vulnificus effect.The highly effective infected wound treatment effect of BG was evaluated on mouse models,revealing significant reduction of local V.vulnificus,accelerated wound contraction,and alleviated inflammation.Besides the high bacterial inhibition of BG,BG remarkably reduced inflammatory response,promoted collagen deposition,neovascularization and re-epithelization,contributing to wound healing.BG is a promising topical biological formulation against infected wounds.
