A novel endoplasmic reticulum stress-related lncRNA prognostic risk model for cutaneous melanoma.

OBJECTIVE: 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). METHODS: 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. RESULTS: 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. CONCLUSION: This study identified a risk model of ten ERS-related lncRNAs that have significant prognostic value in CM and could help guide clinical treatment.

The role of routine laboratory tests after unilateral total knee arthroplasty.

BACKGROUND: Recent studies suggest that routine laboratory tests are not required within 1 day after partial knee arthroplasty. In this study, we evaluated the utility of routine postoperative laboratory tests after initial unilateral total knee arthroplasty (TKA) in an Asian population. In addition, we explored risk factors associated with abnormal test results. METHODS: Clinical data of patients who underwent original unilateral TKA between 2015 and 2020 were retrospectively analyzed. Patient characteristics and laboratory test results were recorded. Multivariate binary logistic regression analysis was performed to identify risk factors associated with 3 abnormal laboratory results. RESULTS: A total of 713 patients, who underwent relevant laboratory tests within 3 days of TKA surgery, were enrolled. Among them, 8.1%, 9.9%, and 3.4% patients with anemia, hypoalbuminemia, and abnormal serum potassium levels required clinical intervention after surgery. Binary logistic regression analysis revealed that preoperative hemoglobin levels, estimated blood loss, and age were independent risk factors of postoperative blood transfusion in TKA patients. On the other hand, preoperative albumin levels, intraoperative blood loss, and operation time were risk factors associated with postoperative albumin supplementation. In addition, lower body mass index (BMI) and preoperative hypokalemia were potential risk factors of postoperative potassium supplementation. CONCLUSION: Considering that more than 90% of abnormal postoperative laboratory tests do not require clinical intervention, we believe that routine laboratory tests after surgery have little significance in patients with primary unilateral TKA. However, postoperative laboratory testing is necessary for patients with established risk factors.

PgtE Enzyme of Salmonella enterica Shares the Similar Biological Roles to Plasminogen Activator (Pla) in Interacting With DEC-205 (CD205), and Enhancing Host Dissemination and Infectivity by Yersinia pestis.

Yersinia pestis, the cause of plague, is a newly evolved Gram-negative bacterium. Through the acquisition of the plasminogen activator (Pla), Y. pestis gained the means to rapidly disseminate throughout its mammalian hosts. It was suggested that Y. pestis utilizes Pla to interact with the DEC-205 (CD205) receptor on antigen-presenting cells (APCs) to initiate host dissemination and infection. However, the evolutionary origin of Pla has not been fully elucidated. The PgtE enzyme of Salmonella enterica, involved in host dissemination, shows sequence similarity with the Y. pestis Pla. In this study, we demonstrated that both Escherichia coli K-12 and Y. pestis bacteria expressing the PgtE-protein were able to interact with primary alveolar macrophages and DEC-205-transfected CHO cells. The interaction between PgtE-expressing bacteria and DEC-205-expressing transfectants could be inhibited by the application of an anti-DEC-205 antibody. Moreover, PgtE-expressing Y. pestis partially re-gained the ability to promote host dissemination and infection. In conclusion, the DEC-205-PgtE interaction plays a role in promoting the dissemination and infection of Y. pestis, suggesting that Pla and the PgtE of S. enterica might share a common evolutionary origin.

[Correlation between pulsed-field gel electrophoresis profiles and Salmonella serotypes].

OBJECTIVE: To analyze the relationship between pulsed-field gel electrophoresis (PFGE)subtyping and serotyping of Salmonella(S.). METHODS: 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. RESULTS: 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. CONCLUSION: 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.

[The fundamental role of stage control technology on the detectability for Salmonella networking laboratory].

OBJECTIVE: To evaluated the fundamental role of stage control technology (SCT) on the detectability for Salmonella networking laboratories. METHODS: 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. RESULTS: 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. CONCLUSION: 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.

[Assessment of the application of variable-number tandem repeat loci of Salmonella Enteritidis in subtyping multiple-locus variable-number tandem repeat analysis].

OBJECTIVE: 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). METHODS: 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. RESULTS: 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. CONCLUSION: 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.

[Typhoid and paratyphoid fever in Yunnan province: distributional patterns and the related meteorological factors].

OBJECTIVE: 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. METHODS: 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. RESULTS: 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)]. CONCLUSION: 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.

[Comparison of the transcriptional levels of mannitol PTS operon between epidemic and non-epidemic strains of Vibrio cholerae].

OBJECTIVE: 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. METHODS: 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. RESULTS: 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. CONCLUSIONS: 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.

[Development and application of real-time polymerase chain reaction to detect Vibrio cholerae O1 and O139 in river water].

OBJECTIVE: 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. METHODS: 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. RESULTS: 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. CONCLUSION: 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.

[Transcriptional repressor gene--mtlR of mannitol PTS operon in Vibrio cholerae].

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.

[Study on infection of different strains of Vibro cholerae O1 by El Tor CTXPhi].

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.

[Surveillance on severe acute respiratory syndrome associated coronavirus in animals at a live animal market of Guangzhou in 2004].

OBJECTIVE: 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. METHODS: 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. RESULTS: 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. CONCLUSION: 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.

[Sequencing and analysis of Vibrio cholerae typing phage VP3 genome].

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.