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Endoplasmic reticulum stress (ERS) and long non-coding RNAs (lncRNAs) are important in melanoma development and progression. This study aimed to explore the prognostic value of ERS-associated lncRNA profiles in cutaneous melanoma (CM).The Cancer Genome Atlas (TCGA) provides the raw data of CM. GSEA website was used to obtain ERS-related genes, and mRNA and LncRNA co-expression network were used to obtain ERS-related lncRNAs. A Lasso regression analysis was used to identify a prognostic risk model for the composition of ERS-related lncRNAs. Patients were divided into high- and low-risk groups based on the model's risk score. The researchers then compared the two groups' survival rates, immune infiltration, chemotherapeutic drug sensitivity, and immune checkpoint gene expression.Thirty-nine ERS-related lncRNAs were discovered to be prognostic. A prognostic risk model made up of ten ERS-related lncRNAs was discovered. Patients in the low-risk group had a better prognosis than those in the high-risk group. An examination of tumor microenvironment revealed that risk scores correlated with immune cell infiltration in eight cases. Dacarbazine, paclitaxel, and cisplatin, three chemotherapy drugs, were more sensitive in the low-risk group than in the high-risk group.This study identified a risk model of ten ERS-related lncRNAs that have significant prognostic value in CM and could help guide clinical treatment.

Typhoid fever remains a severe public health problem in developing countries. The emergence of resistant typhoid, particularly multidrug-resistant typhoid infections, highlights the necessity of monitoring the resistance characteristics of this invasive pathogen. In this study, we report a typhoid fever outbreak caused by multidrug-resistant Salmonella enterica serovar Typhi strains with an ACSSxtT pattern. Resistance genes conferring these phenotypes were harbored by a large conjugative plasmid, which increases the threat of Salmonella Typhi and thus requires close surveillance for dissemination of strains containing such genes.

Vibrio cholerae is an important human pathogen and environmental microflora species that can both propagate in the human intestine and proliferate in zooplankton and aquatic organisms. Cholera is transmitted through food and water. In recent years, outbreaks caused by V. cholerae -contaminated soft-shelled turtles, contaminated mainly with toxigenic serogroup O139, have been frequently reported, posing a new foodborne disease public health problem. In this study, the colonization by toxigenic V. cholerae on the body surfaces and intestines of soft-shelled turtles was explored. Preferred colonization sites on the turtle body surfaces, mainly the carapace and calipash of the dorsal side, were observed for the O139 and O1 strains. Intestinal colonization was also found. The colonization factors of V. cholerae played different roles in the colonization of the soft-shelled turtle's body surface and intestine. Mannose-sensitive hemagglutinin (MSHA) of V. cholerae was necessary for body surface colonization, but no roles were found for toxin-coregulated pili (TCP) or N -acetylglucosamine-binding protein A (GBPA). Both TCP and GBPA play important roles for colonization in the intestine, whereas the deletion of MSHA revealed only a minor colonization-promoting role for this factor. Our study demonstrated that V. cholerae can colonize the surfaces and the intestines of soft-shelled turtles and indicated that the soft-shelled turtles played a role in the transmission of cholera. In addition, this study showed that the soft-shelled turtle has potential value as an animal model in studies of the colonization and environmental adaption mechanisms of V. cholerae in aquatic organisms. IMPORTANCE Cholera is transmitted through water and food. Soft-shelled turtles contaminated with Vibrio cholerae (commonly the serogroup O139 strains) have caused many foodborne infections and outbreaks in recent years, and they have become a foodborne disease problem. Except for epidemiological investigations, no experimental studies have demonstrated the colonization by V. cholerae on soft-shelled turtles. The present studies will benefit our understanding of the interaction between V. cholerae and the soft-shelled turtle. We demonstrated the colonization by V. cholerae on the soft-shelled turtle's body surface and in the intestine and revealed the different roles of major V. cholerae factors for colonization on the body surface and in the intestine. Our work provides experimental evidence for the role of soft-shelled turtles in cholera transmission. In addition, this study also shows the possibility for the soft-shelled turtle to serve as a new animal model for studying the interaction between V. cholerae and aquatic hosts.

OBJECTIVE To identify risk factors for relapse and death in patients with T1 to T2 breast cancer with 0-3 positive axillary lymph nodes. METHODS The case files of 540 breast cancer patients with T1-T2 tumors with 0-3 positive nodes were reviewed retrospectively. Ten-year locoregional recurrence (LRR), distant recurrence (DR), disease-free survival (DFS) and overall survival (OS) of the patients were analyzed. Univariate statistical analysis and Cox proportional hazards models were carried out with SPSS software v.16.0. RESULTS The median follow-up of all the patients was 7.2 years. On multivariate analysis, > 20% positive axillary nodes was the only variable that influenced LRR adversely (hazard ratio[HR], 12.816; 95% confidence interval, 4.657-35.266, P 20% positive axillary nodes and ductal carcinoma were variables that influenced DR adversely (HR, 11.088, 95% confidence interval, 3.807-32.297, P 20% positive axillary nodes were the only variables that had negative effect on 10-year OS (HR, 2.110, 95% confidence interval, 1.364-3.264, P = 0.001; HR, 10.244, 95% confidence interval, 3.497-30.011, P < 0.001) and they were also adverse prognostic variables on 10-year DFS (HR, 1.634, 95% confidence interval, 1.171-2.279, P = 0.004; HR, 7.339, 95% confidence interval, 2.906-18.530, P < 0.001). CONCLUSION Axillary lymph nodal status is the only risk factor with a significant impact on 10-year LRR, DR, OS and DFS. Patients with T1-T2 breast cancer with 0-3 positive lymph nodes have the LRR and DR of over 10 years, and the OS and DFS of less than 10 years, compared to patients with negative lymph nodes. Histology in primary tumors is a significant prognostic factor for the 10-year DR.

To evaluated the fundamental role of stage control technology (SCT) on the detectability for Salmonella networking laboratories.Appropriate Salmonella detection methods after key point control being evaluated, were establishment and optimized. Our training and evaluation networking laboratories participated in the World Health Organization-Global Salmonella Surveillance Project (WHO-GSS) and China-U.S. Collaborative Program on Emerging and Re-emerging infectious diseases Project (GFN) in Shanghai. Staff members from the Yunnan Yuxi city Center for Disease Control and Prevention were trained on Salmonella isolation from diarrhea specimens. Data on annual Salmonella positive rates was collected from the provincial-level monitoring sites to be part of the GSS and GFN projects from 2006 to 2012.The methodology was designed based on the conventional detection procedure of Salmonella which involved the processes as enrichment, isolation, species identification and sero-typing. These methods were simultaneously used to satisfy the sensitivity requirements on non-typhoid Salmonella detection for networking laboratories. Public Health Laboratories in Shanghai had developed from 5 in 2006 to 9 in 2011, and Clinical laboratories from 8 to 22. Number of clinical isolates, including typhoid and non-typhoid Salmonella increased from 196 in 2006 to 1442 in 2011. The positive rate of Salmonella isolated from the clinical diarrhea cases was 2.4% in Yuxi county, in 2012. At present, three other provincial monitoring sites were using the SBG technique as selectivity enrichment broth for Salmonella isolation, with Shanghai having the most stable positive baseline.The method of SCT was proved the premise of the network laboratory construction. Based on this, the improvement of precise phenotypic identification and molecular typing capabilities could reach the level equivalent to the national networking laboratory.

To analyze the relationship between pulsed-field gel electrophoresis (PFGE)subtyping and serotyping of Salmonella(S.).PFGE was performed and profiles were analyzed on 1230 Salmonella isolates which comprising the top five serotypes including Typhimurium, Enteritidis, Derby, Agona and Senftenberg identified in China. The potential predictive relationship between PFGE banding patterns and particular serotypes was compared and the discriminatory consensus band class markers of individual serotypes were identified.Among all the 1230 Salmonella strains, 1149 strains were found assistant with serotyping through PFGE cluster analysis, providing the matching accuracy reaching 93.4%. For the five serotypes, the positive prediction rate appeared more than 90.0% and the negative prediction rate was over 95.0% on serotype cluster prediction.Results presented in this study were representatives of the top 5 Salmonella serovars, showing that PFGE cluster analysis could provide clues to identity and confirmation of serotypes.

To evaluate the feasibility of the application of variable-number tandem repeat (VNTR) loci of Salmonella Enteritidis (S. enteritidis) in subtyping mutiple-locus variable-number tandem repeat analysis (MLVA).A total of 16 isolates of S.enteritidis from different place and time in China were preliminarily assessed by choosing 11 reported VNTR loci, the loci with single amplified bands were picked to subtype all 104 S. enteritidis isolates. The isolates were also analyzed by pulse field gel electrophoresis (PFGE) to compare the superiority or inferiority of MLVA method and PFGE method.Seven VNTR loci were selected from the preliminary screening to expand the analysis, and the 7 VNTR loci had grouped 104 of S.enteritidis isolates into either 16 MLVA subtypes or 22 PFGE subtypes, with the D value at 0.7222 and 0.7974, respectively. Comparing with the isolates in MLVA subtypes, the isolates in PFGE showed a stronger resolving power. Meanwhile the results in PFGE showed a more disperse frequency distribution than those in MLVA.These results indicate that some VNTR locus which have shown a good polymorphism internationally, may fail to show polymorphism in China, thereby, more VNTR loci should be included in MLVA and the wide screening may benefit the unity of global laboratorial methods.

To characterize the spatial distribution of typhoid and paratyphoid fever (TPF) in Yunnan province, China and to determine the effectiveness of meteorological factors on the epidemics of TPF.Data of reported TPF cases in Yunnan province (2001 - 2007) from the China Information System for Diseases Control and Prevention was applied to GIS-based spatial analyses to detect their spatial distribution and clustering of TPF incidence at the county level. Panel data analysis was used to identify the relationships between the TPF incidence and meteorological factors including monthly average temperature, monthly cumulative precipitation and monthly average relative humidity.During the study period, the average incidence of TPF in Yunnan province was 23.11/100 000, with majority of the TPF cases emerged in summer and autumn. Although widely distributed, two TPF clusters were detected in Yunnan province based on the spatial analysis: one area around Yuxi city with the average annual incidence as 207.45/100 000 and another at the junctions of Yunnan province with Burma and Laos. Based on results from panel data analysis, the incidence of TFP was shown to be associated with meteorological factors such as temperature, precipitation, relative humidity and one month lag of temperature increase [10°C increase in the monthly average temperature: IRR = 1.30 (95%CI: 1.24 - 1.36); 10% increase in monthly average relative humidity: IRR = 1.07 (95%CI: 1.05 - 1.09); 100 mm rise in monthly cumulative precipitation: IRR = 1.02 (95%CI: 1.00 - 1.03); and 10°C average temperature increase, the last month: IRR = 1.73 (95%CI: 1.64 - 1.82)].Areas with high TPF incidence were detected in this study, which indicated the key areas for TPF control in Yunnan province. Meteorological factors such as temperature, precipitation and humidity played a role in the incidence of TPF.

To compare the transcription difference of the mannitol PTS genes between epidemic and non-epidemic strains of Vibrio cholerae El Tor in mannitol ferment tests.Growth curves of 10 epidemic strains (slow-ferment) and 10 non-epidemic strains (rapid-ferment) of Vibrio cholerae were detected in the process of fermentation test, and the transcriptional level of mannitol PTS operon of these strains were determined with quantitative reverse-transcriptional PCR.After 4 hours of test, the non-epidemic strains became positive and the average growth density of the non-epidemic strains was higher than that of the epidemic strains; however, some were still lower than the epidemic strains. In contrast, at the eighth hour of test, when epidemic strains got positive, they showed higher average growth density. Compared to the epidemic strains, the transcription of mannitol PTS genes of the non-epidemic strains were much more active at the 1st and 2nd hour and were lower at the 4th and 8th hour.The difference of mannitol PTS operon transcription level should be an important feature to identify the epidemic and non-epidemic strains of Vibrio cholerae, which directly influences the mannitol fermentation rate during the test. The growth rate is not a key factor that affect such difference.

To develop a real-time SYBR Green polymerase chain reaction (PCR) for detection of Vibrio cholerae serogroups O1 and O139, and to evaluate its reliability through detection of estuary water samples.O antigen rfb genes specific for O1 and O139 were used for the design of PCR primers. The real-time SYBR Green PCR system in detecting O1 and O139 specific rfb genes in one tube was developed, and its sensitivity, specificity and reproducibility were evaluated. The ability of the real-time PCR in detection of estuary water samples was compared with the routine PCR and bacteria isolation.The amplification of O1 or O139 specific target gene could be detected according to the melt curve temperature of amplicons. No amplification was observed in the templates of other 10 non-cholerae vibrios. When comparing to the real-time PCR to bacteria isolation in detection of 524 estuary water samples, it showed high sensitivity, plus also positive in real-time PCR detection among all the samples in which bacteria of O1 or O139 were isolated.The real-time SYBR Green PCR could be used as the first step of rapid environment screen of V. cholerae in water samples thus might enhance the efficiency of isolation in screening of large amount of water samples.

The fermentation rates of mannitol in toxigenic and non-toxigenic El Tor strains of Vibrio cholerae are obviously different, which is a valuable indicator in the rapid identification of toxigenic strain. To determine the regulating role of mtlR in transcription of mannitol PTS operon in V. cholerae, and whether it plays a role in the ferment difference of the toxigenic and non-toxigenic strains, the mtlR deletion mutants from the mannitol rapid-ferment strain (non-toxigenic strain) and slow-ferment strain (toxigenic strain) were constructed. Comparisons of the growth in M9 containing 0.2% mannitol as the sole carbon source and pH change in mannitol fermentation media of these wild strains and their mutants, indicated that mtlR is a repressor. Its repression in mtlCBA transcription was further verified with the analyses of quantitative reverse-transcriptional PCR. However, the regulation of mtlR is not the immediate cause of the ferment difference of the toxigenic and non-toxigenic strains. The study also provides the necessary data in the analyses of mannitol ferment difference between the toxigenic and non-toxigenic V. cholerae strains.

DNA sequence and the genome of phage VP3 (a typing phage of V. cholera) were analyzed. A random library of VP3 DNA was constructed by shot-gun library method. The VP3 genome sequence was assembled with contigs sequences, the gaps between different contigs were filled with sequencing data from primer walking. ORFs were predicted; Phylogeny of DNA polymerase sequences was analyzed to determine the class of VP3; The activity of putative promoter genes were analyzed using lacZ report system. VP3 genome is a 39504bp of circular double-stranded DNA. Twenty-seven out of forty-nine putative ORFs were annotated; twenty gene products were homologous with T7-like phages, including DNAP, DNA replicative protein, capsid, tail tubular, tail fiber protein, and DNA packaged protein. The activity of the putative promoter regions was confirmed through cloning those regions to LacZ-fuse plasmid pRS1274 and analysis of the expression of beta galactosidase. The complete genomic sequence of VP3 and phylogenetic tree analysis suggests VP3 is a member of T7 phage family.

To study the horizontal transfer efficiencies of filamentous bacteriophage CTXPhi in different V. cholera O1 strains and the phage immunities of these strains. The infectious El Tor CTXPhi particles genetic marked by chloramphenicol resistance gene were used to infect four different V. cholerae O1 strains in vivo and in vitro. Selected the infected clones based on its character of chloramphenicol resistance and identified and judged the exist form of CTXPhi genome through Southern bolt and other hybridization methods. Calculated the infection rates of different strains and compared each other. Then we analyzed the mechanism of infection and phage immunity. The infection rate of classic strain 1119 with the genetic marked CTX(ET)Phi in vivo is much higher than that in vitro. In vitro experiment, the rate of 1119 is higher than other three El Tor strains. And in El Tor strains, the infection frequency of IEM101 that had no rstR gene is 100 to 1000 times higher than other two strains containing rstR. Classical biotype strain is more susceptible to CTX(ET)Phi particles than El Tor strains. Expression of TCP and the phage immunity mediated by rstR gene affect the horizontal transfection of CTXPhi in V. cholerae strains.

To study the prevalence of severe acute respiratory syndrome coronavirus (SARS-CoV) like virus in animals at a live animal market of Guanzhou in 2004 before and after culling of wild animal action taken by the local authority, in order to predict the re-emerging of SARS from animal originals in this region.Animals at live animal market were sampled for rectal and throat swabs in triplicate. A single step realtime reverse transcription-polymerase chain reaction (RT-PCR) diagnostic kit was performed for screening SARS-CoV like virus, the manual nested RT- PCR and DNA sequencing were performed for confirmation. Only specimens which tested positive for both of the N and P genes by nested RT-PCR were scored as positive.In 31 animals sampled in January 5 2004 before culling of wild animals at Guangdong Province, including 20 cats (Felis catus), 5 red fox (Vulpes vulpes) and 6 Lesser rice field rats (Rattus losea), 8 (25.8%) animals were tested positive for SARS-CoV like virus by RT-PCR methods, of which 4 cats, 3 red fox and one Lesser rice field rats were included. However, two weeks after culling of animals and disinfection of the market were implemented, in 119 animals sampled in January 20 2004, including 6 rabbits (Oryctolagus cuniculus), 13 cats, 46 red jungle fowl (Gallus gallus), 13 spotbill duck (Anas platyrhynchos), 10 greylag goose (Anser anser), 31 Chinese francolin (Franclinus pintadeanus), only rectal swab from one greylag goose was tested positive for SARS-CoV like virus. Furthermore, in 102 animals that including 14 greylag gooses, 3 cats, 5 rabbits, 9 spotbill duck (Anaspoecilorhyncha), 2 Chinese francolin (Franclinus pintadeanus), 8 common pheasant (Phasianus colchicus), 6 pigeons, 9 Chinese muntjac (Muntiacus reevesi), 19 wild boar (Sus scrofa), 16 Lesser rice field rats, 5 dogs, 1 mink (Mustela vison), 3 goats, 2 green peafowl (Pavo muticus) sampled in April, May, June, July, August and November, only rectal swab from one pig was tested positive. However, of 12 and 10 palm civets sampled in November and December including five of which had been at the live animals market for 2 days, none of them was tested positive.This findings revealed that animals being sampled in April, May, June, July, August and November of 2004, only one rectal swab from a pig was tested positive as SARS-CoV like virus, much lower than the results from the previous year, suggesting that the possibility of re-emerging of human infection from animal origins is low for the winter of 2004-2005.