Objective: To establish a rapid detection method for Salmonella based on the combination of enzymatic recombinase amplification (ERA) and lateral flow chromatography (LF), so as to provide technical support for the on-site detection of Salmonella. Methods: Specific ERA primers and probes were designed based on the highly conserved flagella gene fimY in Salmonella. The primers were screened using capillary electrophoresis, and the probes were designed according to the amplification range of the screened primers. The amplification temperature and time were optimized to establish the amplification method, and the product was detected using LF strips. A standard strain of Salmonella was used to verify the sensitivity, 10 other gut bacteria were used to to verify the specificity and sensitivity, and the nucleic acid of the actual Salmonella strains was amplified to verify the detectability. Results: After screening for Salmonella-specific primers using capillary electrophoresis, the minimum detection concentration was 5 copies/μL under the amplification temperature of 37 ℃ and reaction time of 20 minutes. This method had a positive amplification result for Salmonella nucleic acid, and the amplification results of 10 other gut bacteria were all negative, with good specificity. Conclusion This method provides a possibility for on-site point of care testing of Salmonella infection.

Objective To investigate the associations between single nucleotide polymorphisms in the gene encoding human leukocyte antigen-G ( HLA-G) and the genetic susceptibility to high risk human papillomavirus ( HPV) type 18 infection in the subjects from Taizhou, Zhejiang province. Methods The genetic polymorphisms of HLA-G gene (14 bp In/Del and +3142C/G) in cervical samples collected from HPV 18-positive and healthy women were analyzed by PCR and gene sequencing technology. Statistical anal-ysis was performed by using SPSS version 16. 0. Chi-squared test was used to analyze the differences with HLA-G gene allele and genotype frequencies between healthy subjects and patients. Results Compared with healthy subjects, women with oncogenic HPV18 infection showed lower frequencies of -14 bp allele,-14 bp/-14 bp genotype and -14 bp/+3142C haplotype (P<0. 05). Moreover, lower frequencies of+3142C/C genotype were detected in HPV18-infected women with pathologically normal cervix (6. 3% vs 21. 1%, OR=0. 25, P<0. 05) and higher percentages of +3142G/G genotype were detected in women with CIN2/3 stage HPV18 infection as compared with those of the control group (68. 8% vs 35. 1%, OR=4. 06, P<0. 05). Conclusion The HLA-G gene 3′ UTR polymorphisms were closely associated with HPV18 in-fection and cervical intraepithelial neoplasia.

Objective To characterize Hfq-dependent phenotypes in stress response and to dissect Hfq-dependent transcription of virulence genes and stress-responsive genes in Vibrio cholera.Methods The hfq null mutant strain (△hfq) and the complemented mutant strain (△hfq/pUC18-hfq) were constructed from the wild-type Vibrio cholera.Comparisons on the motility,biofilm formation,growth under various oxygen-supplying conditions,outer membrane resistance,and sensitivity to oxidative stress were analyzed between the wild type strain and the mutant strains.Reverse-transcript fluorescence quantitative PCR (RT-qPCR) was used to determine the transcriptional levels of target genes in the above mentioned strains.Results △hfq and △hfq/pUC18-hfq strains were successfully constructed.The motility,outer membrane resistance and sensitivity to oxidative stress were reduced,but biofilm formation was enhanced in the hfq null mutant strain.RT-qPCR testified that Hfq had regulation effects on gene transcription for forming falagellum,extracellular polysaccharide,outer membrane protein and oxidative stress in Vibrio cholera.Conclusion As a RNA chaperone,Hfq could affect Vibrio cholera in its biofilm formation,resistance to oxidative stress and antibiotics resistance through regulating the transcription of multiple metabolic genes and virulence genes,which indicates that Hfq,combined with other regulators,may play a key role in the complex regulation of metabolic genes and virulence genes.

Objective To investigate the cytolytic activity of extracellular cytolytic toxin rVVC of Vibrio vulnificus on the apoptosis of human ECV304 cells, and to analyze the activities of Caspase-3,-8 and -9. Methods The cytotoxic effect of refolded rVVC on the growth and apoptosis of ECV304 cells was identified by MTT, Hochest33342/PI fluorescent staining, flow cytometry and DNA agarose electrophoresis analysis, respectively. The activities of Caspase-3, -8 and -9 was measured using a colorimetric method. Results The viability of human ECV304 cells exposed to rVVC was inhibited by rVVC after 24 h. 2.0 HU/ml rVVC groups had a better cytotoxic effect to human ECV304 than that of 0.5 HU /ml rVVC groups. The apoptosis of human ECV304 cells in 2.0 HU/ml rVVC+40 μmol/L Z-VAD-FMK groups was relative reduced than that of 2.0 HU/ml of rVVC groups. After 0.5 h treatment with 2.0 HU/ml of rVVC, the Caspase-3 activity in human ECV304 cells increased gradually and reached the peak at 3 h (versus control groups, P<0.01). The activity of Caspase-8 and -9 remained unchanging. Conclusion The rVVC has cytotoxic effect on human ECV304 and the cytolysin is probably correlated with Caspase-3.

Objective To analyze the expression of peripheral blood CD13+CD4+CD25hi regulatory T cells (Treg cells) in patients with diffuse large B-cell lymphoma (DLBCL) and its clinical significance. Methods The expression of peripheral blood CD13+CD4+CD25hi Treg cells in 58 newly diagnosed patients with DLBCL and 30 healthy adults was detected by flow cytometry, and the relationship between its expression and the clinical indicators were analyzed statistically. Results The levels of peripheral blood CD13+CD4+CD25hi Treg cells in newly diagnosed DLBCL and healthy adults were different, with statistically significant difference [(36.37 ±11.89) % vs. (9.03 ±2.10) %, t = 7.168, P < 0.001]. The level of peripheral blood CD13+CD4+CD25hi Treg cells was significantly higher in patients with IPI score 3ˉ5 than that in patients with IPI score 0ˉ2[(44.28±10.10)%vs. (21.51±6.23)%, t=ˉ9.347, P=0.03]. The expression of peripheral blood CD13+ CD4+ CD25hi Treg cells in stages Ⅱ, Ⅲ and Ⅳ patients were (19.48 ±1.34) %, (33.98 ±8.03) % and (47.89±8.25) %respectively, and there were significant differences among three groups (F= 38.363, P<0.001). The levels of peripheral blood CD13+CD4+CD25hi Treg cells had no relationship with age, sex or LDH level (all P>0.05). Conclusion The levels of peripheral blood CD13+CD4+CD25hi Treg cells are higher in DLBCL patients, which has a close relationship between the expression of CD13+CD4+CD25hi Treg cells and clinical stage and prognosis.

Objective To evaluate the clinical effectiveness of early-stage keep warm intervention on improving low temperature risk children treated with blood purification (BP). Methods Ninety children were randomized into observation group (46 cases) and control group of (44 cases) from July 2013 to September 2015. Control group were nursed with conventional BP standard operation process, while the observation group were nursed additionally with heat insulation blanket before 30 min of booting machine, and recorded the central body temperature of 0 min,30 min,60 min,90 min,120 min for each 60 min 1 time in the future. Until the end of the blood purification 60 min. Low temperature complications were recorded and judged between the two groups. Results In the observation group, 107 cases of low body temperature occurred during the course of 7 cases of blood purification, occurrence rate of 6.5% (7/107). The control group was 15.8% (16/101). The difference between the 2 groups was statistically significant (χ2=4.569,P<0.05);In the continuous blood purification group, observation group 10.6% (5/47), The incidence rate of control group was 29.3% (12/41), he difference between the 2 groups was statistically significant (χ2=4.876,P<0.05). Conclusions Application of heat insulation blanket at the early-stage may effectively reduce the risk of hypothermia complications in the children treated with blood purification. The continuous blood purification effect is more significant.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Diarrhea ; virology ; Female ; Humans ; Male ; Middle Aged ; Oligonucleotide Array Sequence Analysis ; methods ; Young Adult

s: The onset of Crohn’s disease (CD) is the interaction of environment, heredity, infection, immune and other factors. It is also closely related to abnormal immune functions. Without special treatment, CD is identified as a modern refractory disease. By syndrome differentiation and treatment, traditional Chinese medicine (TCM) can effectively relieve disease conditions, improve the quality of life and reduce side effects of modern medication. The core compatibility ofBai-Zhu andFu-Ling can reinforce spleen-qi and dispel dampness, which met the common pathogenesis of CD. Therefore, the combination is comprehensively used in the compound prescription. Our previous study found thatBai-Zhu Fu-Ling decoctioncan reduce the vasoactive intestinal peptide (VIP) of animal model of spleen-qi deficiency, downregulate VIP receptors, decrease the affinity of VIP receptors and improve animal model’s sIgA. To further clarify the effects about neurotransmitters and their correlation with the immune system in the pathogenesis of CD and the intervention mechanism treated by different proportional compatibility ofBai-Zhu Fu-Ling decoction, we studied influences of the decoction on related transmitters of nerve- immune network and functions of receptors, as well as cytokine secretion and signal transduction of TLR4-NF-κB. Our studies can provide references and foundations to further explore TCM treatment of CD.

AIM: To study the pathological change in mouse organs immunitied by inactivated SARS-CoV vaccine. METHODS: Inactivated SARS-CoV vaccine was injected into BALB/c and C57BL/6 mice. Anti-SARS antibody was analyzed by ELISA. After 8 weeks, the immunitied mice were killed and those organs were analyzed by pathological methods. RESULTS: Anti-SARS antibody in mice was positive after 8 days. Only minimal injury was observed in a few lungs and livers, but the other organs were not. CONCLUSIONS: Inactivated SARS-CoV vaccine induced mice to create antibody, whereas they did not cause severe injury. This result will be valuable for vaccine into clinical research. [

OBJECTIVETo determine the function of twin-arginine translocation system (Tat) and gene cluster in Vibrio strains and to analyze the homology of tat gene cluster among different Vibrio spp. strains based on N16961 and tatABC mutant strains N169-dtat.

METHODSDifferent serotypes of biotype strains of Vibrio spp. were selected to detect the transcription of 4 genes of Tat transport system and upstream ubi aarF gene and downstream cyt551 gene by the total RNA reverse transcription and homologicity of the gene cluster by sequencing analysis.

RESULTSOur results showed that the 4 genes of tat cluster (tatA, tatB, tatC, and tatE) were intragenic and co-transcribed. We found that ubi aarF gene could be co-transcribed with tatA, tatB, but not with tatC. The electron transport chain and energy metabolism-related genes, cytochrome C551 peroxidase gene, and 4 genes located at upstream of tatABC operon were not transcribed with tatABC. Although the co-transcription between ubi aarF and tatAB was blocked in N169-dtat strain, they were still transcribed separately. Homologous analysis of genes of tat cluster in different types of Vibrio cholerae showed that tat gene cluster was a very conservative.

CONCLUSIONThe ubi and aarF gene might be co-transcribed with genes of tat cluster in Vibrio cholerae, which and the close relationship showed that they might play a key function in Vibrio cholerae.