Increased 5-hydroxymethylcytosine and Ten-eleven Translocation Protein Expression in Ultraviolet B-irradiated HaCaT Cells

Background： DNA hydroxymethylation refers to a chemical modification process in which 5-methylcytosine （5mC） is catalyzed to 5-hydroxymethylcytosine （5hmC） by ten-eleven translocation （TET） family proteins. Recent studies have revealed that aberrant TETs expression or 5hmC level may play important roles in the occurrence and development of various pathological and physiological processes including cancer and aging. This study aimed to explore the relation between aberrant DNA hydroxymethylation with skin photoaging and to investigate the levels of TETs, 5mC, and 5hmC expression 24 h after 40 mJ/cm^2 and 80 mJ/cm^2 doses of ultraviolet B （UVB） irradiation to HaCaT cells. Methods： To explore whether aberrant DNA hydroxymethylation is also related to skin photoaging, 40 mJ/cm^2 and 80 mJ/cm^2 doses of UVB were chosen to treat keratinocytes （HaCaT cells）. After 24 h of UVB irradiation, 5mC and 5hmC levels were determined by immunohistochemistry （IHC） and immunofluorescence （IF）, and at the same time, the expression levels of matrix metalloproteinase 1 （MMP-1） and TETs were assessed by reverse transcription-polymerase chain reaction or Western blot analysis. Results： After 40 mJ/cm^2 and 80 mJ/cm^2 doses of UVB exposure, both IHC and IF results showed that 5hmC levels increased significantly, while the 5mC levels did not exhibit significant changes in HaCaT cells, compared with HaCat cells without UVB exposure. Moreover, compared with HaCat cells without UVB exposure, the levels ofTET1, TET2, and TET3 mRNA and protein expression were significantly upregulated （mRNA： P = 0.0022 and 0.0043 for TET1; all P 〈 0.0001 for TET2; all P = 0.0006 for TET3; protein： P = 0.0012 and 0.0006 tbr TET 1 ; all P = 0.0022 for TET2; and all P = 0.0002 for TET3）, and the levels of MMP- 1 mRNA expression increased dose dependently in 40 mJ/cm^2 and 80 mJ/cm^2 UVB-irradiated groups. Conclusion： UVB radiation could cause increased 5hmC and TET expression, which might become a novel biomarker in UVB-related skin aging.

Multiple Functions of Ten-eleven Translocation 1 during Tumorigenesis

Objective： Aberrant expression of ten-eleven translocation 1 （TET1） plays a critical role in tumor development and progression. We systematically summarized the latest research progress on the role and mechanisms of TET1 in cancer biology. Data Sources： Relevant articles published in English from 1980 to April 2016 were selected from the PubMed database. Tile terms ＂＇ten-eleven translocation 1,＂ ＂＇SmC,＂ ＇5hmC,＂ ＇microRNA,＇＂ ＂hypoxia,＂ and ＂＇embryonic stem cell＇＂ were used tbr the search. Study Selection： Articles focusing on the role and mechanism ofTETI in tumor were reviewed, including clinical and basic research articles. Results： TET proteins, the key enzymes converting 5-methylcytosine to 5-hydroxymethylcytosine, play vital roles in DNA demethylation regulation. Recent studies have shown that loss of TETI is associated with tumorigenesis and can be used as a potential biomarker for cancer therapy, which indicates that TETI serves as tumor suppressor gene. Moreover, besides its dioxygenase activity, TET1 could induce epithelial-mesenchymal transition and act as a coactivator to regulate gene transcription, such as developmental regulator in embryonic stem cells （ESCs） and hypoxia-responsive gene in cancer. The regulation of TETl is also correlated with microRNA in a posttranscriptional modification process. Hence, it is complex but critical to coaprehend the mechanisms of TETI in the biology of ESCs and cancer. Conclusions： TET I not only serves as a demethylation enzyme but also plays multiple roles during tumorigenesis and progression. More studies should be carried out to elucidate the exact mechanisms of TETI and its associations with cancer betbre considering it as a therapeutic tool.

Optimized silicon fertilization regime weakens cadmium translocation and increases its biotransformation in rice tissues

In acidic paddy fields of South China,rice(Oryza sativa L.)faces the dual challenges of cadmium(Cd)toxicity and silicon(Si)deficiency.Although previous studies have highlighted the functions of Si application timing and strategies in mitigating Cd-stressed rice,the precise mechanisms underlying the health restoration of Cd-toxic rice and the assurance of grain safety remain elusive.This study explored Cd translocation and detoxification in the shoots of rice regulated by various Si fertilization regimes:Si(T)(all Si added before transplanting),Si(J)(all Si added at jointing),and Si(TJ)(half Si added both before transplanting and at jointing).The findings revealed that the regime of Si(TJ)was more beneficial to rice health and grain safety than Si(T)and Si(J).The osmotic regulators such as proline,soluble sugars,and soluble proteins were significantly boosted by Si(TJ)compared to other Si treatments,and which enhanced membrane integrity,balanced intracellular pH,and increased Cd tolerance of rice.Furthermore,Si(TJ)was more effective than Si(T)and Si(J)on the Cd sequestration in the cell wall,Cd bio-passivation,and the down-regulated expression of the Cd transport genes.The concentrations of Cd in the xylem and phloem treated with Si(TJ)were reduced significantly.Additionally,Si(TJ)facilitated much more Cd bound with the outer layer proteins of grains,and promoted Cd chelation and complexation by phytic acid,phenolics,and flavonoids.Overall,Si(TJ)outperformed Si(T)and Si(J)in harmonizing the phycological processes,inhibiting Cd translocation,and enhancing Cd detoxification in rice plant.Thereby the split Si application strategy offers potential for reducing Cd toxicity in rice grain.

Adipsin inhibits Irak2 mitochondrial translocation and improves fatty acid β-oxidation to alleviate diabetic cardiomyopathy

Background Diabetic cardiomyopathy (DCM) causes the myocardium to rely on fatty acid β-oxidation for energy. The accumulation of intracellular lipids and fatty acids in the myocardium usually results in lipotoxicity, which impairs myocardial function. Adipsin may play an important protective role in the pathogenesis of DCM. The aim of this study is to investigate the regulatory effect of Adipsin on DCM lipotoxicity and its molecular mechanism.MethodsA high-fat diet (HFD)-induced type 2 diabetes mellitus model was constructed in mice with adipose tissue-specific overexpression of Adipsin (Adipsin-Tg). Liquid chromatography-tandem mass spectrometry (LC–MS/MS), glutathione-S-transferase (GST) pull-down technique, Co-immunoprecipitation (Co-IP) and immunofluorescence colocalization analyses were used to investigate the molecules which can directly interact with Adipsin. The immunocolloidal gold method was also used to detect the interaction between Adipsin and its downstream modulator.ResultsThe expression of Adipsin was significantly downregulated in the HFD-induced DCM model (P < 0.05). Adipose tissue-specific overexpression of Adipsin significantly improved cardiac function and alleviated cardiac remodeling in DCM (P < 0.05). Adipsin overexpression also alleviated mitochondrial oxidative phosphorylation function in diabetic stress (P < 0.05). LC–MS/MS analysis, GST pull-down technique and Co-IP studies revealed that interleukin-1 receptor-associated kinase-like 2 (Irak2) was a downstream regulator of Adipsin. Immunofluorescence analysis also revealed that Adipsin was co-localized with Irak2 in cardiomyocytes. Immunocolloidal gold electron microscopy and Western blotting analysis indicated that Adipsin inhibited the mitochondrial translocation of Irak2 in DCM, thus dampening the interaction between Irak2 and prohibitin (Phb)-optic atrophy protein 1 (Opa1) on mitochondria and improving the structural integrity and function of mitochondria (P < 0.05). Interestingly, in the presence of Irak2 knockdown, Adipsin overexpression did not further alleviate myocardial mitochondrial destruction and cardiac dysfunction, suggesting a downstream role of Irak2 in Adipsin-induced responses (P < 0.05). Consistent with these findings, overexpression of Adipsin after Irak2 knockdown did not further reduce the accumulation of lipids and their metabolites in the cardiac myocardium, nor did it enhance the oxidation capacity of cardiomyocytes expose to palmitate (PA) (P < 0.05). These results indicated that Irak2 may be a downstream regulator of Adipsin.ConclusionsAdipsin improves fatty acid β-oxidation and alleviates mitochondrial injury in DCM. The mechanism is related to Irak2 interaction and inhibition of Irak2 mitochondrial translocation.

Ten-eleven translocation-2 affects the fate of cells and has therapeutic potential in digestive tumors

Ten-eleven translocation(TET)methylcytosine dioxygenases catalyze the oxidative reactions of 5-methylcytosine(5-mC)to 5-hydroxymethylcytosine(5-hmC),5-formylcytosine(5-fC),and 5-carboxylcytosine(5-caC),which are intermediate steps during DNA demethylation.It is reported that somatic mutations of TET2 gene are identified in a variety of human tumors,especially in hematological malignancies.The tendency and mechanism of cellular differentiation in different systems are affected by TET2 via regulation of associated gene expression or maintenance of demethylated state.TET2 acts as a critical driver of tumorigenesis through the conversion of 5-mC to 5-hmC and successive oxidation products.Sometimes,it requires special interactions and cofactors.Here,we reviewed recent advances in understanding the function of TET2 proteins in regulating cell differentiation,and its role in various tumors focusing on several digestive cancers.

Effects of Slow-release Nitrogen on Dry Matter Accumulation,Translocation and Yield of Summer Maize

[Objectives]This study was conducted to investigate the effects of slow-release nitrogen fertilizer on dry matter accumulation and translocation of summer maize.[Methods]With Zhoudan 9 as the test variety,six different treatment were set up:blank control(CK1),slow-release urea 75 kg/hm^(2)(C1),slow-release urea 150 kg/hm^(2)(C2),slow-release urea 225 kg/hm^(2)(C3),slow-release urea 300 kg/hm^(2)(C4)and ordinary urea 300 kg/hm^(2)(CK2),to study the change law of dry matter accumulation and translocation in summer maize.[Results]Treatment slow-release urea 225 kg/hm^(2)(C4)showed summer maize yield,dry matter translocation between organs,grain contribution rate and proportion of grain dry matter in the full ripe stage higher than other treatments.Considering the weight loss and cost factors,slow-release urea 225 kg/hm^(2)(C3)could be recommended as the fertilizing amount for summer maize.[Conclusions]This study provides theoretical reference for rational selection of fertilizers for reducing fertilizer application and increasing fertilizer efficiency,and for production of summer maize in Shajiang black soil region.

Identification of Triticum aestivum-Haynaldia villosa Translocation Line T6BS·6BL-2VS

[Objective] The aim of experiment was to provide a new germplasm for wheat breeding by further using desirable genes in 2V chromosome of Haynaldia villosa.[Method] Through hybridization between common wheat(Triticum aestivum)-Haynaldia villosa disomic substitution line and common wheat Nonglin26-3C chromosome of Aegilops triuncialis disomic addition line,the analysis methods such as chromosome C-banding,genomic in situ hybridization and molecular marker technique were comprehensively applied and combined characters investigation.[Result] The wheat-Haynaldia villosa translocation line(T6BS·6BL-2VS)was selected from hybrid progenies to conduct characters investigation,which found some bristles on glume ridge of T6BS·6BL-2VS.[Conclusion] The translocation line induced by gametocidal chromosome was a small segment translocation line and the gene of bristle on glume ridge of Haynaldia villosa was located between the middle and the terminal of 2VS.

Fruit Photosynthesis and Assimilate Translocation and Partitioning: Their Characteristics and Role in Sugar Accumulation in Developing Citrus unshiu Fruit

Dynamics of dry- or fresh-weight of fruit, peel photosynthetic rate and chlorophyll content, and the characteristics of translocation and distribution of radiolabelled assimilates from leaf or fruit were examined in developing satsuma mandarin (Citrus unshiu Marc. cv. Miyagawa wase) fruit from primary stage of fruit enlargement up to fruit full ripe. Change in fruit photosynthetic rate was some what related to the change in the chlorophyll content of peel. Fruit photosynthetic rate markedly declined as chlorophyll degradation occurred in the peel. Before full ripe stage of the fruit, photosynthates produced by a 14C-fed leaf were mainly distributed to juice sacs even during periods when dry matter accumulation in peel was more rapid than that in juice sacs. At the full ripe stage, peel photosynthetic rate approached zero and peel became the major sink of leaf photosynthates. Most of the peel assimilates, however, remained in situ for up to 48 h after feeding 14CO 2 to the fruit, only a small portion being transported to other parts of fruit. The percentage of fruit photosynthates exported decreased with fruit development and ripening, but the peak rate of export to juice sacs amount to as high as 12%. The sugar content and dry weights of peel and juice sacs in shaded fruit were lower than that in the control fruit. These results show that peel assimilate was mainly consumed in peel respiration and growth and thus the dependence on leaf photosynthates decreased. Part of this assimiate was used in sugar accumulation in juice sacs of fruit.

Translocation and Distribution of Imidacloprid in Tobacco with Two Application Methods

[Objective] To compare the translocation and distribution of imidacloprid in tobacco with spray and root irrigation application methods. [Methods] Pot experiment in the greenhouse was carried out, and LC-MSMS was used to determine the con- tent of imidacloprid in different parts of tobacco plants （roots, stems, the upper, middle and lower leaves） at different time. [Results] The imidacloprid could be absorbed by root and could be transported to all parts of the tobacco plant after irrigating root, but the original deposition amount was larger and the transport efficiency was lower after spraying. [Conclusion] The translocation and distribution of imidacloprid by spraying was more uniform and the holding efficiency was better, but imidacloprid with root irrigation could act on leaf directly, and had better readily availability.

Polymorphisms of CMYA3 Gene in 13/17 Robertsonian Translocation Pigs

[Objective] The aim was to study the polymorphism of CMYA3 gene in the 148 pigs of hybrid offspring of 13/17 Robertsonian translocation pigs [2n = 37,rob （13;17）] intercrossing.[Method] PCR-RFLP method was adopted.[Result] A 507 bp fragment of CMYA3 gene was obtained by PCR amplification,and then amplification product by using restriction nuclease Bsh1236Ⅰ was detected by agarose gel electrophoresis.As a result,both alleles （A and B） of the loci were found in the population.The frequencies of allele A and B were 0.699 and 0.301.The genotype frequencies of AA,AB and BB were 0.615,0.169 and 0.216.The frequencies of allele A and genotype AA were significantly higher than allele B and genotype BB in populations.[Conclusion] The study will provide theoretical basis for molecular breeding and marker-assisted selection of 13/17 Robertsonian translocation pigs.

Meiotic Behavior of 1BL/1RS Translocation Chromosome and Alien Chromosome in Two Tri-genera Hybrids

The behavior of wheat-rye translocation chromosome and alien chromosome including Thinopyrum and Haynaldia chromosome at meiosis was investigated in two hybrids by fluorescence in situ hybridization (FISH). Misdivision of translocation chromosome at anaphase I and rye chromatin micronucleus at tetrad stage were observed, A plant with one normal 1BL/1RS translocation chromosome and one 1BL/1RS translocation chromosome deleted about 1/3 of rye chromosome arm in length was identified. One plant with wheat-Thinopyrum non-Robertson translocation chromosome was also detected in the F-2 population of Yi4212 x Yi4095. That could be the results of unequal misdivision of wheat-rye 1BL/1RS translocation chromosome and Thinopyrum chromosome during meiosis. No interaction between translocation chromosome and alien chromosome at meiosis was supported by the data of the distribution frequencies of translocation chromosome and Thinopyrum or Haynaldia chromosome in the progeny of two hybrids. The results may be useful to cultivate new germplasms with different length of rye 1R short arm and wheat-alien non-Robertson translocation tines under wheat background.

基因组Translocation排序问题的改进多项式算法

该文给出基因组 Translocation排序问题的一个改进多项式算法 .原算法所用存储空间为 O(n) ,时间复杂度为 O(n3) .文中改进算法仍采用 O(n)存储空间 ,时间复杂度为 O(n2 logn) .具体地 ,将计算 Translocation距离的时间复杂度由 O(n3)改进为 O(n2 ) ,将计算 Translocation序列的时间复杂度由 O(n3)改进为 O(n2 logn) .

Effect of Interaction upon Translocation of Confined Polymer Chain Through Nanopore

The effect of the interaction between nanopore and chain monomer on the translocation of a single polymer chain confined in a finite size square through an interacting nanopore to a large space has been studied by two-dimensional bond fluctuation model with Monte Carlo simulation. Results indicate that the free energy barrier before the successful translocation of the chain depends linearly on the chain length as well as the nanopore length for different pore-polymer interaction, and the attractive interaction reduces the free energy barrier, leading to the reduction of the average trapping time.

染色体转位(Translocation)与癌症

许多种癌症是由染色体转位引起的。这是因为转位使原癌基因活化并过量表达 ,或是转位使基因融合。大部分已确认的转位都与免疫系统 (免疫球蛋白和T细胞受体基因 )本身固有的V(D)J重组有关。此外 ,外界因素 ,如电离辐射和某些化学药物等因素使DNA双链断裂 ,也可能导致染色体转位和癌变。了解染色体转位和癌变机制 。

Microbiota and the gut-liver axis:Bacterial translocation,inflammation and infection in cirrhosis

Liver disease is associated with qualitative and quantitative changes in the intestinal microbiota. In cirrhotic patients the alteration in gut microbiota is characterized by an overgrowth of potentially pathogenic bacteria (i.e., gram negative species) and a decrease in autochthonous familiae. Here we summarize the available literature on the risk of gut dysbiosis in liver cirrhosis and its clinical consequences. We therefore described the features of the complex interaction between gut microbiota and cirrhotic host, the so called &#x0201c;gut-liver axis&#x0201d;, with a particular attention to the acquired risk of bacterial translocation, systemic inflammation and the relationship with systemic infections in the cirrhotic patient. Such knowledge might help to develop novel and innovative strategies for the prevention and therapy of gut dysbiosis and its complication in liver cirrhosis.

Effect of artesunate supplementation on bacterial translocation and dysbiosis of gut microbiota in rats with liver cirrhosis

AIM: To evaluate the effect of artesunate(AS) supplementation on bacterial translocation(BT) and gut microbiota in a rat model of liver cirrhosis. METHODS: Fifty-four male Sprague-Dawley rats were randomly divided into a normal control group(N), a liver cirrhosis group(M) and a liver cirrhosis group intervened with AS(MA). Each group was sampled at 4, 6 and 8 wk. Liver cirrhosis was induced by injection of carbon tetrachloride(CCl4), intragastric administration of 10% ethanol, and feeding a high fat diet. Rats in the MA group were intragastrically administered with AS(25 mg/kg body weight, once daily). Injuries of the liver and intestinal mucosa were assessed by hematoxylineosin or Masson's trichrome staining. Liver index was calculated as a ratio of the organ weight(g) to body weight(g). The gut microbiota was examined by automated ribosomal intergenic-spacer analysis of fecal DNA. BT was assessed by standard microbiological techniques in the blood, mesenteric lymph nodes(MLNs), liver, spleen, and kidney. RESULTS: Compared to group N, the body weight was reduced significantly in groups M and MA due to the development of liver cirrhosis over the period of 8 wk. The body weight was higher in group MA than in group M. The liver indices were significantly elevated at 4, 6 and 8 wk in groups M and MA compared to group N. AS supplementation partially decreased the liver indices in group MA. Marked histopathologic changes in the liver and small intestinal mucosa in group M were observed, which were alleviated in group MA. Levels of pro-inflammatory interleukin-6 and tumor necrosis factor-α were significantly elevated at 8 wk in ileal homogenates in group M compared to group N, which were decreased after AS supplementation in group MA. The dysbiosis of gut microbiota indicated by the mean diversity(Shannon index) and mean similarity(Sorenson index) was severe as the liver cirrhosis developed, and AS supplementation had an apparent intervention effect on the dysbiosis of gut microbiota at 4 wk. The occurrence of BT was increased in the liver of group M compared to that of group N. AS supplementation reduced BT in group MA at 8 wk. BT also occurred in the MLNs, spleen, and kidney, which was reduced by AS supplementation. BT was not detected in the blood in any group.CONCLUSION: Dysbiosis of gut microbiota, injury of intestinal mucosal barrier and BT occurred as liver cirrhosis progressed, which might enhance inflammation and aggravate liver injury. AS may have other nonantimalarial effects that modulate gut microbiota,inhibit BT and alleviate inflammation, resulting in a reduction in CCl4, alcohol and high fat-caused damages to the liver and intestine.

Relationship between enteric microecologic dysbiosis and bacterial translocation in acute necrotizing pancreatitis

AIM To investigate the potential role of intestinal microflora barrier in the pathogenesis of pancreatic infection. METHODS Fifteen dogs were colonized with a strain of E.coli JM109 bearing ampicillin resistance plasmid PUC18. The animals were divided into two groups. In experimental group ( n =8), acute necrotizing pancreatitis (ANP) was induced by injection of 0 5ml/kg of sodium tarocholate with 3000U/kg trypsin into the pancreatic duct. The control group ( n =7) underwent laparotomy only. All animals were sacrificed 7 days later. Mucosal and luminal microflora of intestine were analyzed quantitatively, and various organs were harvested for culturing, blood samples were obtained for determination of serum amylase activities and plasma lipopolysaccharide (LPS) concentrations. RESULTS In the experimental group, the number of E.coli in the intestine was much higher than those of the controls, while bifidobacterium and lactobacillus were decreased significantly (jejunum, 1 75±0 95 vs 2 35±0 79, P <0 05; 1 13±0 8 vs 1 83±0 64, P <0 05; ileum, 2 89±0 86 vs 3 87±1 05, P <0 05; 1 78±0 79 vs 3 79±1 11, P <0 01; cecum, 2 70±0 88 vs 4 89±0 87, P <001; 2 81±0 73 vs 3 24±0 84, P <0 05. Content of cecum, 3 06±0 87 vs 5 15±1 44, P <0 01; 2 67±0 61 vs 4 25±0 81, P <0 01), resulting in reversal of bifido bacterium/ E.coli ratio as compared with the control group (jejunum, 0 51±0 76 vs 1 23±0 53, P <0 05; ileum, 0 62±0 68 vs 1 16±0 32, P <0 05; cecum, 0 46±0 44 vs 1 03±0 64, P <0 05). In addition, intestinal bacteria were isolated from organs of all animals in the experimental group, and JM109 was also detected in most cases. Positive blood culture was 75 0% and 62 5% on day 1 and 2 after induction of ANP, respectively, but no bacterium was found in the controls. As compared with the control group, blood LPS levels and serum amylase activities increased 1-3 times and 3-8 times respectively. CONCLUSION Microecological disturbance could occur in ANP, and overgrowth of intestinal gram negative bacteria may lead to translocation to the pancreas and other organs, becoming the source of pancreatic and peripancreatic infection.

Influence of splanchnic vascular infusion on the content of endotoxins in plasma and the translocation of intestinal bacteria in rats with acute hemorrhage necrosis pancreatitis

The main reason for the death of the patient with acute hemorrhage necrosis pancreatitis (AHNP) is pancreatic infection and multi-organ failure caused by endotoxemia and intestinal bacterial translocation[1-7]. However, the pathogenesis of endotoxemia and intestinal bacterial translocation remains a question[8-10]; moreover, no effective method of prevention and cure for it has been found till now[11 -15] In the present study, we infused low dose dopamine and low molecular weight dextran through the catheters to abdominal aorta and portal vein, and observed its influence on the endotoxin concentration in plasma and the rate of translocation of intestinal bacteria in AHNP rats.

Bacterial translocation and changes in the intestinal microbiome associated with alcoholic liver disease

Alcoholic liver disease progresses through several stages of tissue damage, from simple steatosis to alcoholic hepatitis, fibrosis, or cirrhosis. Alcohol also affects the intestine, increases intestinal permeability and changes the bacterial microflora. Liver disease severity correlates with levels of systemic bacterial products in patients, and experimental alcoholic liver disease is dependent on gut derived bacterial products in mice. Supporting evidence for the importance of bacterial translocation comes from animal studies demonstrating that intestinal decontamination is associated with decreased liver fibrogenesis. In addition, mice with a gene mutation or deletion encoding receptors for either bacterial products or signaling molecules downstream from these receptors, are resistant to alcohol-induced liver disease. Despite this strong association, the exact molecular mechanism of bacterial translocation and of how changes in the intestinal microbiome contribute to liver disease progression remains largely unknown. In this review we will summarize evidence for bacterial translocation and enteric microbial changes in response to alcoholic liver injury and chronic alcoholic liver disease. We will further describe consequences of intestinal dysbiosis on host biology. We finally discuss how therapeutic interventions may modify the gastrointestinal microflora and prevent or reduce alcoholic liver disease progression.

Tracing method study of bacterial translocation in vivo

INTRODUCTIONEndogenesis infection plays an important role innosocomial infection.By studying progress ofbacteria translocation from intestinal tract,theconcept of gut-origin infection has been acceptedgradually.Because of no ideal tracing method,there were some controversies.In order to solve theproblem,the PUC19 plasmid vector tracing