[Etiological analysis on a foodborne disease outbreak caused by two serotypes of Salmonella].

Objective: To understand the etiological characteristics of 2 serotypes of Salmonella strains isolated from a foodborne disease outbreak. Methods: A total of 11 anal swabs of the cases, 13 suspected contaminated food and 10 environmental samples were collected from a foodborne disease outbreak occurred on September 8, 2022 in a school. The anal swabs were enriched with selenite brilliant green enrichment broth (SBG) and brain heart infusion broth (BHI) respectively. PCR detection and culture of common intestinal pathogens were carried out. The suspected food samples were tested according to national standards for food safety. Multiple suspected Salmonella colonies were obtained and selected for serotype determination and whole genome sequencing. Serotypes were determined based on the whole-genome sequence, and clustering analysis was performed based on core genome single nucleotide polymorphism (SNP). Results: The positive rates of Salmonella in anal swabs and suspected food samples were 9/11 and 5/13 respectively. Both Salmonella Uganda and Salmonella Idikan were isolated from 4 anal swabs and 4 suspected food samples. For the remaining samples, only Salmonella Uganda or Salmonella Idikan was isolated in each sample. The positive rate of Salmonella in 11 anal swabs of the cases after BHI enrichment for 12 h and 24 h were all 9/11 in real-time PCR, same to the culture results. Salmonella Uganda and Salmonella Idikan formed two independent and genetically distant lineages in the clustering tree based on core genome SNP, and 0-14 and 0-23 SNP were observed in Salmonella Uganda and Salmonella Idikan respectively. Conclusions: This foodborne disease outbreak was probably caused by Salmonella Uganda and Salmonella Idikan, which both exhibited strong genetic diversity. The PCR based pathogen screening strategy plus pathogen enrichment for cases' annal swabs can be used in the routine outbreak investigation.

[Drug resistance and genomic characteristics of a strain of O139 Vibrio cholerae isolated from human bloodstream infection].

Objective: To analyze the drug resistance and genomic characteristics of a strain of serogroup O139 Vibrio cholerae producing cholera toxin isolated from the bloodstream of a person with bacteremia. Methods: The broth dilution method and automatic drug sensitivity analyzer were used to determine the antibiotic sensitivity of the strain. The complete genome sequence of the strain was obtained by using second-generation gene sequencing and nanopore sequencing. BLAST software was used for comparison and analysis with CARD, Resfinder, ISfinder, VFDB, and other databases. The drug-resistant genes, insertion sequences and virulence genes carried by the strain were identified. MEGA 5.1 software was used to construct a genetic phylogenetic tree based on the core genomic single nucleotide polymorphisms. Results: V. cholerae SH400, as the toxigenic strain, carried multiple virulence-related genes and four virulence islands. The strain was resistant to streptomycin, tetracycline and cotrimoxazole, carrying corresponding drug-resistant genes. The strain also carried IncA/C plasmid with the size of 172914 bp and contained 10 drug-resistant genes. Combined with the genomic evolutionary relationship, this study found that the drug-resistant genes and drug-resistant plasmids carried among strains showed certain aggregation. The traditional ST type of strain SH400 was ST69, and the cgMLST type was a new type highly similar to cgST-252. Conclusion: This strain of serogroup O139 V. cholerae carries the ctxAB gene, multiple drug-resistant genes and IncA/C plasmid, and there are multiple drug-resistant islands.

[Lithocholic acid decreases mRNA stability of nuclear receptor PPARα by upregulating miR-21 expression in hepatoma HepG2 cells].

OBJECTIVE: To investigate the regulatory effects of lithocholic acid (LCA) on nuclear receptor peroxisome proliferatoractivated receptor-alpha (PPARα) mRNA stability at the post-transcriptional level. METHODS: PPARα 3'UTR luciferase reporter gene vectors were constructed and transfected into HepG2 cells to observe the changes in cellular luciferase activity in response to LCA treatments. Bioinformatic prediction and miRNA PCR array technique were used to identify the differentially expressed miRNAs induced by LCA and their potential binding sites on the 3'UTR. The binding sites (Mut1, Mut2 and Mut1+Mut2) were mutated to compare the changes in cellular luciferase activity following LCA treatment. Western blotting and RTqPCR were used to detect the activated signaling pathway and the expression levels of its downstream transcription factors in LCA-treated cells. The changes in PPARα protein expression level were detected in the cells following overexpression of the transcription factors. RESULTS: Treatment with 100 µmol/L LCA significantly reduced luciferase activity of PPARα 3'UTR1 and 3'UTR2 in HepG2 cells by more than 50% (P<0.01) and induced significant upregulation of miR-21 and miR-22, especially the former (by 2.35 folds, P<0.05). Two predicted miR-21-binding sites in the 3'UTR1 were mutated to construct Mut1, Mut2 and Mut1+Mut2 reporter gene vectors. LCA treatment down-regulated 3'UTR1 luciferase activity by 51%, while Mut1, Mut2, and Mut1+Mut2 were down-regulated by 37%, 39%, and 13%, respectively. LCA caused ERK1/2 phosphorylation and activation of the ERK1/2 signaling pathway, and treatment with 100 µmol/L LCA upregulated the expression of transcription factor early growth response 1 (EGR1) by 5.83 folds (P<0.01). Transient overexpression of EGR1 significantly decreased cellular PPARα protein levels (P<0.05). CONCLUSION: LCA reduces PPARα mRNA stability and thus decreases PPARα mRNA and protein expressions in hepatocytes by activating the ERK1/2 signaling pathway and upregulating EGR1 and miR-21, which targets 3'UTR regulatory region of PPARα mRNA.

[Construction of spvD gene deletion mutant and compensation strains in Salmonella enteritidis and its effects on Caco-2 cells].

Objective: To analyze the effects of spvD gene on invasion and intracellular proliferation of Caco-2 cells and in order to provide insight into the function of that gene and the underlying mechanism of Salmonella caused infection. Methods: Functional verification of spvD gene deletion mutant and compensation strain. The deletion mutant strain was constructed through a suicide plasmid-mediated homologous recombination. The compensation plasmid constructed by cloning the coding sequence of spvD by PCR into plasmid pBAD33 was mobilized into the deletion mutant by conjugation and the pBAD33 was introduced into wild strains and deleted mutant strains as control. The relative expression of spvD mRNA was detected by quantitative reverse transcription PCR. In order to analyze the virulence of spvD against Caco-2 cells, Caco-2 cells was cocultured with wild type Salmonella enteritidis carrying spvD gene, the deletion mutant strain and compensation strain respectively. The expression level of spvD mRNA and the the number of Salmonella enteritidis after Caco-2 cells intervention were compared between the three groups by LSD-t test, and the invasion rate was compared by χ2 test. Results: The expression level of spvD mRNA in wild type Salmonella enteritidis was set as unit "1", the deletion mutant strain was "0.00", and the compensation strain was "2.60" (LSD-twild, deleted=1.11, P=0.31; LSD-twild, compensation=-1.77, P=0.13; LSD-t deleted, compensation=-2.88, P=0.03), which confirmed the successful construction of the deletion mutant strain and the compensation strain. The invasion experiment results of the above three Salmonella enteritidis strains on Caco-2 cells showed that the invasion rate of wild strain was 0.23%, the invasion rate of deleted mutant strain was 0.16%, and the invasion rate of compensation strain was 0.16%, with no statistical significance (χ2=1.13, P=0.570). By comparing the number of Salmonella enteritidis at different time points after Caco-2 cells intervention, it was discovered that the number of Salmonella enteritidis in wild strains (6.50×106 CFU/ml) and compensation strains (7.25×106 CFU/ml) was significantly increased than that in deletion mutant strain (1.90×106 CFU/ml) after 16 h coculture (LSD-twild, deleted=7.95, P=0.00; LSD-twild, compensation=-1.27, P=0.25; LSD-t deleted, compensation=-9.22, P=0.00). Conclusion: It is not considered that spvD gene can affect the invasion of Salmonella enteritidis on Caco-2 cells, but the gene can promote the reproduction of Salmonella enteritidis in Caco-2 cells.

[Comparison and application of postprandial and fasting blood lipid levels in 839 physical examinees in Beijing].

Objective: To investigate the feasibility of application of non-fasting dyslipidemia cutoff values in community population. Methods: Self-control study was used. 839 physical examinees (292 males and 547 females) were recruited in clinical laboratory of Guang'an men Hospital from January to October 2018. The median (interquartile range) of age was 60 (54, 66) years. Blood samples were collected before and at 4 h after a standard breakfast. Comparison of fasting and postprandial lipoprotein levels was performed using Paired-Samples T Test or Two-Related-Samples Wilcoxon. The changes of 4-hour postprandial blood lipid levels and the percentages of postprandial dyslipidemia according to different stratification of fasting dyslipidemia were performed using one-way ANOVA and χ2 test, respectively. Results: Compared with fasting, 4-hour postprandial total cholesterol (TC), low density lipoprotein cholesterol (LDL-C), high density lipoprotein cholesterol (HDL-C), non-high density lipoprotein cholesterol (non-HDL-C), apolipoprotein A1 (ApoA1) and apolipoprotein B (ApoB) decreased slightly, postprandial triglyceride (TG) increased by 0.72 mmol/L, and postprandial remnant-like lipoprotein cholesterol (RLP-C) increased by 0.27 mmol/L (t or Z values = 10.26,22.94,24.22,4.71,16.61,26.92,-23.58,-19.35, P<0.05, respectively). According to the non-fasting dyslipidemia cut-off values recommended by the European consensus, there were 10%, 16.6%, 10.1%, 12.3%, 30% and 34.9% of the population in the appropriate levels of fasting TC, LDL-C, HDL-C, non-HDL-C, TG and RLP-C distributed in elevated levels of postprandial, respectively. The changes of 4-hour postprandial TC, LDL-C, non-HDL-C and HDL-C increased with the elevation of fasting level (F=9.50,6.18,8.07,3.86,P<0.01), and the maximum changes of TC≤3.5%, LDL-C≤6.8%, non-HDL-C≤2.9%, HDL-C≤6.3%; the change of 4-hour postprandial TG increased slightly first and then decreased significantly (51.3% vs. 57.9% vs. 39.2%, F=19.05, P<0.01); the change of 4-hour postprandial RLP-C decreased (50.8% vs. 33.2%, F=10.40, P<0.01). The cut-off values of 4-hour postprandial dyslipidemia were TC ≥5.1 mmol/L, LDL-C ≥3.2 mmol/L, HDL-C ≤0.9 mmol/L, non-HDL-C ≥4.0 mmol/L and RLP-C ≥1.0 mmol/L. The cut-off values of borderline elevated and elevated TG levels were ≥2.2 mmol/L and ≥3.4 mmol/L, respectively. Conclusions: The cut-off values of postprandial dyslipidemia including TC, LDL-C, HDL-C, non-HDL-C and RLP-C were preliminarily established in community population, which could be applied to the routine lipid profile evaluation in the physical examination population. And it might be needed that postprandial TG was managed hierarchically according to different cut-off values.

Probiotic and prebiotic interventions for non-alcoholic fatty liver disease: a systematic review and network meta-analysis.

Many studies have associated altered intestinal bacterial communities and non-alcoholic fatty liver disease, but the putative effects are inconclusive. The purpose of this network meta-analysis (NMA) was to evaluate the effects of probiotics, prebiotics, and synbiotics on non-alcoholic fatty liver disease through randomised intervention trials. Literature searches were performed until March 2020. For each outcome, a random NMA was performed, the surface under the cumulative ranking curve (SUCRA) was determined. A total of 22 randomised trials comparing prebiotic, probiotic, and synbiotic treatments included 1301 participants. Considering all seven results (aspartate aminotransferase, alanine aminotransferase, body mass index, weight, total cholesterol, high-density lipoprotein cholesterol, and low-density lipoprotein cholesterol) together, the highest SUCRA values are probiotics (94%), synbiotics (61%) and prebiotics (56%), respectively. NMA results provide evidence that probiotics, prebiotics, and synbiotics can alleviate non-alcoholic fatty liver disease. However, due to the lack of high-quality randomised trials, this research also has some limitations.

Obesity Measures and Definitions of Sarcopenic Obesity in Singaporean Adults - the Yishun Study.

OBJECTIVES: Due to the lack of a uniform obesity definition, there is marked variability in reported sarcopenic obesity (SO) prevalence and associated health outcomes. We compare the association of SO with physical function using current Asian Working Group for Sarcopenia (AWGS) guidelines and different obesity measures to propose the most optimal SO diagnostic formulation according to functional impairment, and describe SO prevalence among community-dwelling young and old adults. DESIGN: Obesity was defined according to waist circumference (WC), percentage body fat (PBF), fat mass index (FMI), fat mass/fat-free mass ratio (FM/FFM), or body mass index (BMI). SO was defined as the presence of both obesity and AWGS sarcopenia. Muscle function was compared among phenotypes and obesity definitions using ANOVA. Differences across obesity measures were further ascertained using multiple linear regressions to determine their associations with the Short Physical Performance Battery (SPPB). SETTING: Community-dwelling adults 21 years old and above were recruited from a large urban residential town in Singapore. PARTICIPANTS: 535 community-dwelling Singaporeans were recruited (21-90 years old, 57.9% women), filling quotas of 20-40 participants in each sex- and age-group. MEASUREMENTS: We took measurements of height, weight, BMI, waist and hip circumferences, body fat, muscle mass, muscle strength, and functional assessments. Questionnaire-based physical and cognitive factors were also assessed. RESULTS: Overall prevalence of SO was 7.6% (WC-based), 5.1% (PBF-based), 2.7% (FMI-based), 1.5% (FM/FFM-based), and 0.4% (BMI-based). SO was significantly associated with SPPB only in the FMI model (p<0.05), and total variance explained by the different regression models was highest for the FMI model. CONCLUSIONS: Our findings suggest FMI as the most preferred measure for obesity and support its use as a diagnostic criteria for SO.

Juvenile hormone regulates the reproductive diapause through Methoprene-tolerant gene in Galeruca daurica.

Juvenile hormone (JH) signalling plays an important role in regulation of reproductive diapause in insects. However, its underlying molecular mechanism has been unclear. Methoprene-tolerant (Met), as a universal JH receptor, is involved in JH action. To gain some insight into its function in the reproductive diapause of Galeruca daurica, a serious pest on the Inner Mongolia grasslands undergoing obligatory summer diapause at the adult stage, we cloned the complete open-reading frame (ORF) sequences of Met and other 7 JH signalling-related genes, including JH acid methyltransferase (JHAMT), JH esterase (JHE), JH epoxide hydrolase (JHEH), Krüppel homologue 1 (Kr-h1), vitellogenin (Vg), forkhead box O (FOXO) and fatty acid synthase 2 (FAS2), from this species. GdMet encoded a putative protein, which contained three domains typical of the bHLH-PAS family. Expression patterns of these eight genes were developmentally regulated during adult development. Topical application of JH analogue (JHA) methoprene into the 3-day-old and 5-day-old adults induced the expression of GdMet. Silencing GdMet by RNAi inhibited the expression of JHBP, JHE, Kr-h1 and Vg, whereas promoted the FAS2 expression, which enhanced lipid accumulation and fat body development, and finally induced the adults into diapause ahead. Combining with our previous results, we conclude that JH may regulate reproductive diapause through a conserved Met-dependent pathway in G. daurica.

Effect of meat temperature on moisture loss, water properties, and protein profiles of broiler pectoralis major with the woody breast condition.

The objective of this study was to investigate the influence of meat temperature on moisture loss, muscle water properties, and protein profiles in broiler pectoralis major with the severe woody breast (WB) condition. Broiler breast samples were collected from a commercial plant and sorted into normal, WB, and pale, soft, and exudative (PSE). Temperature treatments included 23°C, 40°C, 53°C, 57°C, 68°C, and 90°C based on denaturation of major muscle protein types during heating. Moisture loss was estimated with weight changes, water properties were measured with time-domain nuclear magnetic resonance measurements, and protein profiles were determined with SDS electrophoresis gel. There were no differences in moisture loss between 3 groups at meat temperature 23°C, 40°C, and 57°C. Moisture loss of WB samples was greater than normal and PSE at either 68°C or 90°C; however, it was the least at 53°C. Only close changing trend was noted between the intramyofibrillar water (T21) reduction and moisture loss. The extramyofibrillar (T22) water content and reduction in WB meat during heating were consistent greater, and electrophoretic profiles differed among 3 muscle conditions. Data suggest that greater reductions in intramyofibrillar and extramyofibrillar water are responsible for the increased moisture loss in WB meat at higher temperature.

Malnutrition and Sarcopenia in Community-Dwelling Adults in Singapore: Yishun Health Study.

OBJECTIVES: To determine the overlapping prevalence of malnutrition and sarcopenia and the association between parameters of malnutrition with muscle mass and strength in a community-dwelling Singaporean adult population. DESIGN: This was a cross-sectional study. SETTING: Large north-eastern residential town of Yishun in Singapore. PARTICIPANTS: Random sampling of community-dwelling Singaporeans aged 21-90 years old (n=541). MEASUREMENTS: Anthropometry, body composition and handgrip strength (muscle strength) were measured. Sarcopenia was identified using dual-energy x-ray absorptiometry scan (muscle mass). Nutritional status was measured using Mini Nutritional Assessment (MNA-SF). Other questionnaires collected included physical activity and cognition. Associations between nutritional status with sarcopenia as well as with muscle mass and strength were analysed using multinomial logistics and linear regressions. RESULTS: The overall population-adjusted prevalence of those at nutritional risk and malnourished were 18.5% and 0.1% respectively. More than a third of participants (35%) who were at nutritional risk were sarcopenic. Malnourished participants were all sarcopenic (100%, N=2) whereas those who were sarcopenic, 27.0% (N=37) were at nutritional risk/malnourished. Being at nutritional risk/malnourished was significantly associated with 2 to 3 times increased odds of sarcopenia in multivariate analyses adjusting for age, gender, physical activity level and cognition, and fat mass index. Favourable MNA parameter scores on food intake and BMI were positively associated with greater muscle mass and handgrip strength (p<0.05). CONCLUSION: Given the overlapping clinical presentation of malnutrition and sarcopenia, community screening protocols should include combination screening of nutritional status and sarcopenia with appropriate interventions to mitigate risk of adverse health outcomes.

Research Note: Comparison of 3 methods used for estimating cook loss in broiler breast meat.

The objective of this study was to evaluate different methods used for estimating cook loss in broiler breast fillets (pectoralis major). Two experiments were conducted. In the first experiment, intact fillets were weighed, cooked to 75°C, and then subjected to 1 of 3 postcook handling treatments: cooling for 5 min at room temperature before reweighing (5-minute), cooling at room temperature until they reached room temperature before reweighing (RT), or cooling in ice water until they reached room temperature before reweighing (IW). In the second experiment, breast fillet portions were used to compare the effects of endpoint temperature (53°C, 57°C, 68°C, 75°C, or 90°C) on cook loss estimation by the 5-minute and RT methods. Breast fillets were collected from local chicken processing plants and trimmed to similar weight prior to cooking. Cook loss, cook loss retention, and total cook loss after 24 h in cooked fillets were measured for comparisons. Data showed that cook loss (<17%) and total loss (19.3%) estimated with the IW method were lower (P < 0.05) than those with the 5-minute and RT methods (19-21% for cook loss and 21.1-21.3% for total loss), which did not differ from each other. When the endpoint temperature was ≥75°C, no differences in cook loss estimates or moisture loss were noted between the 5 min and RT methods (after 3 h cooling). However, when the temperatures were 53°C to 75°C, cook loss estimations were significantly different (P < 0.05) between the 5 min and RT methods (more than 4%). Reduced endpoint temperature resulted in increasing differences (from less than 5% to more than 9%) in cook loss estimates. These results demonstrate that endpoint cooking temperature and postcooking sample handling methods may affect cook loss estimates in broiler breast meat.

Muscle water properties in raw intact broiler breast fillets with the woody breast condition.

The purpose of this study was to investigate the relationships between muscle water properties, water-holding capacity (WHC), and woody breast (WB) severity in intact raw broiler breast fillets. Broiler pectoralis major deboned at 3 h postmortem was collected from a commercial plant and categorized as normal (NORM), moderate WB, or severe WB (SEV). Meat drip loss was calculated based on weight loss during overnight storage at 4°C. Water properties of the intact fillets were determined with time domain nuclear magnetic resonance and the T2 relaxation times were determined using an inverse Laplace algorithm (CONTIN). Three T2 water components, hydration water (T2b), intra-myofibrillar water (T21), and extra-myofibrillar water (T22), were identified. With increasing WB severity, the time constant of each water component and the relative content of T22 (P22) increased while the relative areas of T2b and T21 (P2b and P21, respectively) decreased. Spearman correlation analysis showed that there were significant correlations between the WB condition score and either the time constant or normalized area for each T2 component. T22 normalized areas (A22) were most strongly correlated with the WB score (r = 0.75); however, the weakest correlation was found between the WB score and T21 areas (A21). Pearson correlation analysis revealed that the strongest correlation (r = 0.64) was found between A22 and drip loss; however, there was no correlation between A21 and drip loss. Within the NORM group, drip loss was significantly correlated to the time constants for both T2b and T21. Within the SEV group, only A22 was significantly correlated to drip loss. These data indicate that the WB condition has a significant impact on the distribution of water within the intact muscle tissue. The content of extra-myofibrillar water in broiler breast fillets may be a key factor responsible for the poor WHC measurements in WB meat.

Relationships between instrumental texture measurements and subjective woody breast condition scores in raw broiler breast fillets.

The objective of this study was to compare the relationships between instrumental texture measurements and subjective woody breast (WB) scores in raw broiler breast fillets. A total of 181 broiler breast fillets were scored based on palpable hardness and rigidity ranging from 1.0 to 3.0 in 0.5 increments. Texture properties of raw fillets were measured with 3 different instrumental methods: compression force, blunt Meullenet-Owens Razor Shear (BMORS), and Meullenet-Owens Razor Shear (MORS). Compression force was measured based on % of fillet height (30%) and distance (10 mm). Blunt Meullenet-Owens Razor Shear and MORS measurements included peak force, energy, and peak counts. One-way ANOVA of instrumental texture measurements were performed. Spearman correlations between WB scores and instrumental texture measurements and Pearson correlations between 3 instrumental measurements were analyzed. ANOVA results showed that the best means separations between WB scores were found with the compression method. The weakest means separations were observed with MORS force and BMORS peak counts. Spearman correlation coefficients showed that there were significant relationships between WB scores and instrumental measurements. The strongest correlations were found between subjective WB scores and compression force measurements (r = 0.58-0.73, P < 0.0001), followed by BMORS force and energy (r = 0.55-0.56, P < 0 0.0001), MORS energy and peak count (r = 0.47-0.50, P < 0 0.0001), and BMORS peak count (r = -0.18, P = 0.015). The weakest correlation was found between the WB scores and MORS force (r = 0.17, P = 0.023). Pearson correlation coefficients between 3 different instrumental texture methods were also significant (P < 0.0001). These results demonstrate that there are significant correlations between subjective WB scores and instrumental texture measurements but that correlation strength varies with the instrumental method. Instrumental texture measurements can be used as references for subjective WB scores. Compression force method has the best potential for assessing subjective WB condition scores.

[Genomic recombination of the vibrio cholerae serogroup O1 El Tor pandemic strains].

Objective: To analyze the genomic recombination of the vibrio cholerae serogroup O1 El Tor pandemic strains. Methods: A total of 292 complete or draft genome sequences of Vibrio cholerae O1 serogroup El Tor strains isolated from 1937 to 2015 were selected from National Biotechnology Information Center database. The genome alignment of strains was computed by snippy software by using N16961 as reference sequence. Then ClonalFrameML software was used to do the recombinant analysis. The wilcox.test function in agricolae package was used to compare the number recombinant segments and the total length of recombinant regions between small and large chromosomes. The kruskal function was used to compare the number recombinant segments and the total length of recombinant regions among different isolation continents. The KOBAS tool was used to do the gene ontology enrichment analysis of recombinant hotspot genes. Results: Of all 292 strains of Vibrio cholerae, 163 strains (55.8%) were recombined. The median of normalized recombinant segment number of small chromosome was 4.7×10(-6) (9.3×10(-7), 2.0×10(-5)), which was significantly larger than that of large chromosome [2.4×10(-6) (3.4×10(-7), 5.7×10(-6))] (P<0.001). The median (P(25),P(75)) of recombinant segment number of strains isolated from Africa, Asia, Europe, North America and South America were 23(1.0,33.0), 1.0(0.0,34.0), 6.0(2.0,13.0), 0.0(0.0,1.0) and 29.5(6.8,56.8), respectively, and the difference was statistically significant (P<0.001). The median (P(25),P(75)) of total length of recombinant regions of strains isolated from Africa, Asia, Europe, North America and South America were 233.0(4.0, 461.0), 11.0(0.0, 695.5), 56.0(4.0,111.0), 0.0(0.0,9.0) and 347.5(132.8,1 323.5) bp, respectively, and the difference was statistically significant (P<0.001). Gene ontology Enrichment analysis showed that the functions of 62 recombinant hotspot genes were mainly enrichment in chemotaxis, taxis, response to external stimulus, receptor activity and molecular transducer activity. Conclustion: In this study, we found that there were significant differences in the number of recombinant fragments and the length of recombinant regions between large and small chromosomes of Vibrio cholerae El Tor. We also found significant differences in the number of recombinant fragments and the total length of recombinant regions among different continents.

[Patients with anti-GABA (B) receptor encephalitis: 8 cases report].

Objective: To discuss the clinical features, diagnosis, therapy and prognosis of anti-GABA (B) receptor encephalitis. Methods: Clinical data of patients with anti-gaba B receptor encephalitis admitted to the department of neurology of the second hospital of hebei medical university from April 2016 to February 2018 were collected and retrospectively analyzed. Results: Eight patients with anti-GABA (B) receptor encephalitis were included. All the eight cases presented with epileptic seizures. The initial symptoms were clinical seizures in six cases, memory deficit in one case, mental and behavior disorder in one case. Cerebrospinal fluid (CSF) test indicated high intracranial pressure in four patients, increased protein in two patients and lymphocytosis in four patients. Long-term electroencephalogram (EEG) showed abnormal in all patients. Brain MRI showed limbic system lesions in four patients. The anti-GABA (B) receptor antibody was positive in both serum and CSF in all patients. Four patients had small cell lung cancer. All the eight patients received immunotherapy and symptoms were improved. After 6-24 months' follow-up, 1 patient recovered completely, and in the remaining 7 patients, mild symptoms were left behind. Conclusions: The anti-GABA (B) receptor encephalitis is characterized by refractory epilepsy. Epilepsy is the most common initial symptom. About 50% patients are accompanied by tumors. Anti-GABA (B) receptor antibodies testing and tumor screening should be actively carried out.

[Prediction of syncope with nonlinear dynamic analysis during head-up tilt in vasovagal syncope patients].

OBJECTIVE: To quantify the relationship between cerebral blood flow velocity and peripheral blood pressure during hypotension period, aiming to predict the brain hypotension before symptomatic occurrence. METHODS: Twenty vasovagal syncope (VVS) patients who had a previous clinical history were selected in groups and 20 pair-matched control subjects underwent 70° tilt-up test. The subjects remained supine for 30 minutes before recordings when Doppler probes, electrodes and Finapres device were prepared. After continuous baseline recordings for 10 min, the subjects underwent head up tilt (HUT) test (70°), and were standing upright for 30 minutes or until syncope was imminent. For ethical reasons, the subjects were turned back to supine position immediately after SBP dropped to ≥20 mmHg, when their consciousness persisted. The point of syncope was synchronized for all the subjects by the point SBP reached the minima. Their beat-to-beat blood pressures (BP) were recorded continuously and bilateral middle cerebral artery (MCA) flow velocities were obtained with two 2 MHz Doppler probes from a transcranial Doppler ultrasonography (TCD) system. A nonlinear dynamic method--multimodal pressure flow (MMPF) analysis was introduced to access cerebral autoregulation during different time intervals. We introduced a new indicator--syncope index (SI), which was extracted from blood flow velocity (BFV) signal to evaluate the variation of cerebral vascular tension, and could reflect the deepness of dicrotic notch in BFV signal. RESULTS: Compared with the syncope index of the baseline value at the beginning of the tilt test, SI in VVS group showed significantly lower when the VVS occurred (0.16±0.10 vs.0.27±0.10,P<0.01),while there was no significant difference in syncope index between the control group at the end of the tilt test and the baseline value at the beginning of the tilt test. For those VVS patients, pulse index and resistance index had no significant change. Syncope index decreased significantly 3 minutes before the point of syncope (0.23±0.07 vs.0.29±0.07,P<0.01). CONCLUSION: Dynamic regulation is exhausted when vasovagal syncope occurred. Tension decrease of small vessels could have some relationship with loss of the cerebral autoregulation capability. The proposed syncope index could be a useful parameter in predicting syncope of VVS patients since it decreased significantly up to 3 minutes earlier from the point of syncope.

[Predictive factors for lymph node metastasis in patients with poorly differentiated early gastric cancer].

Objective: This study aimed to identify clinicopathological factors predictive of lymph node metastasis in patients with the poorly differentiated early gastric cancer (EGC) to assess the feasibility of using endoscopic submucosal dissection (ESD). Methods: The records of patients with poorly differentiated early gastric cancer undergoing gastric radical resection between January 2012 and December 2016 were reviewed in Ruijin hospital. Those with distant metastasis, two or more malignant tumors, remnant gastric cancer, neo adjuvant therapy, previous history of gastric surgery or clear history of perigastric lymphadenectomy, and mixed tumors were excluded. Age, sex, presence of ulcerous lesion, tumor size, tumor location, depth of invasion, type of differentiation, lymphatic vessel invasion, vascular invasion, nerve invasion and HER2 expression were collected. Univariate and multivariate stepwise logistic regression analyses were used to identify the independent risk factors of perigastric lymph node metastasis.According to the Guidelines for the Treatment of Gastric Cancer (2018 edition) of the Chinese Society of Clinical Oncology (CSCO), the expanded indications of ESD for EGC are as follows: (1)no ulcerative lesions, the maximum diameter of lesions >2 cm of differentiated intramucosal cancer; (2)ulcerative lesions, the maximum diameter of lesions ≤3 cm of differentiated intramucosal cancer; (3)no ulcerative lesions, undifferentiated intramucosal carcinoma with diameter ≤2 cm. The relationship between clinicopathological factors and lymph node metastasis was analyzed. Results: A total of 517 patients, aged 21-83 (57.1±11.7), including 307 males and 210 females, were enrolled in the study. Among them, 114 (22.0%) patients had lymph node metastasis. Univariate analysis showed that ulcerative lesion (P=0.042), tumor diameter (P=0.048), depth of invasion (P<0.001), location of tumors (P<0.001), lymphatic vessel invasion (P=0.009), vascular invasion (P<0.001) and nerve invasion (P=0.028) were related to lymph node metastasis after radical resection of poorly differentiated early adenocarcinoma. Age, sex, type of differentiation and HER2 expression were not significantly correlated to lymph node metastasis (P>0.05). Multivariate analysis showed that tumor size (OR=1.61, 95% CI: 1.03-2.52, P=0.037), depth of invasion (OR=2.77, 95% CI:1.66-4.63, P<0.001), lymphatic duct invasion (OR=14.74, 95% CI:1.58-137.36, P=0.018) were independent risk factors for lymph node metastasis in poorly differentiated EGC, and ulcerative lesion was not a risk factor for lymph node metastasis (OR=0.82, 95% CI:0.56-1.18,P=0.285). A total of 119 patients with poorly differentiated EGC fully complied with the relative indications of ESD recommended by the Japanese Statute and the criteria for radical resection after ESD. Among them, 14 (11.8%) still had perigastric lymph node metastasis, while the gender, tumor diameter, location, differentiation and HER2 expression were not associated with lymph node metastasis (P>0.05). Conclusion: For patients with poorly differentiated EGC, the application of ESD should be carefully weighed with precise assessment of tumor diameter, depth of invasion, and lymphatic duct invasion.

Effects of Consecutive Versus Non-consecutive Days of Resistance Training on Strength, Body Composition, and Red Blood Cells.

Health authorities worldwide recommend 2-3 days per week of resistance training (RT) performed ∼48-72 h apart. However, the influence of recovery period between RT sessions on muscle strength, body composition, and red blood cells (RBCs) are unclear. Aim: Examine the effects of three consecutive (C) or non-consecutive (NC) days of RT per week for 12 weeks on strength, body composition, and RBCs. Methods: Thirty young, healthy and recreationally active males were randomly assigned to 3 C (∼24 h between sessions) or NC (∼48-72 h between sessions) days of RT per week for 12 weeks. Both groups performed three sets of 10 repetitions at 10-repetition maximum (RM) of leg press, latissimus pulldown, leg curl, shoulder press, and leg extension for each session. Ten RM and body composition were assessed pre- and post-RT. RBC parameters were measured on the first session before RT, and 0 and 24 h post-3rd session in untrained (week 1) and trained (week 12) states. Results: No training × group interaction was found for all strength and body composition parameters (p = 0.075-0.974). Training increased strength for all exercises, bone mineral density, and total body mass via increased lean and bone mass (p < 0.001). There was no interaction (p = 0.076-0.994) and RT induced temporal changes in all RBC parameters (p < 0.001-0.003) except RBC corrected for plasma volume changes (time × training interaction; p = 0.001). Training increased hematocrit and lowered mean corpuscular hemoglobin and mean corpuscular hemoglobin concentration (p = 0.001-0.041) but did not alter uncorrected RBC, hemoglobin, mean corpuscular volume and RBC distribution width (p = 0.178-0.797). Conclusion: Both C and NC RT induced similar improvements in strength and body composition, and changes in RBC parameters.

Neutron flux measurements on a mock-up of a storage cask for high-level nuclear waste using 2.5 MeV neutrons.

To store and dispose spent nuclear fuel, shielding casks are employed to reduce the emitted radiation. To evaluate the exposure of employees handling such casks, Monte Carlo radiation transport codes can be employed. Nevertheless, to assess the reliability of these codes and nuclear data, experimental checks are required. In this study, a neutron generator (NG) producing neutrons of 2.5 MeV was employed to simulate neutrons produced in spent nuclear fuel. Different configurations of shielding layers of steel and polyethylene were positioned between the target of the NG and a NE-213 detector. The results of the measurements of neutron and γ radiation and the corresponding simulations with the code MCNP6 are presented. Details of the experimental set-up as well as neutron and photon flux spectra are provided as reference points for such NG investigations with shielding structures.

Reinfection hazard of hand-foot-mouth disease in Wuhan, China, using Cox-proportional hazard model.

Hand-foot-mouth disease (HFMD) is an acute infectious disease caused by serotypes of the enterovirus (EV) family. HFMD reinfection occurs commonly in lack of cross-protection between different EV serotypes. In this study, we investigated the hazards of HFMD reinfection using Cox-proportional hazard model. Retrospective data of 95 209 HFMD cases in Wuhan during 2008-2015 was used. Kaplan-Meier survival methods and Cox-proportional hazard model were used to estimate the hazard probabilities. Of the all HFMD cases, about 2% experienced reinfection (1842/95 209). Kaplan-Meier curves revealed the reinfection risk sharply increased before 40 months from first infection. Higher hazards of reinfection were detected among those who were males, aged 3 years and below, scattered children, belonging to urban areas and first infected with coxsackievirus (CV)-A16 compared with their respective counterparts. Cox-proportional hazard model suggested that gender, age, group, living area and serotypes of first infection had significant effect on reinfection even after adjusting for potential confounding effects of other selected factors considered in the study. These results indicate that boys aged 3 years and below, especially those living in urban areas and first infected with CV-A16 are more prone to reinfection. Interventions should be imposed on these high-risk populations.