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@#Abstract: Objective To investigate the molecular characteristics and drug resistance of non-O1/non-O139 Vibrio cholerae in Zhongshan City, and to provide laboratory basis for cholera prevention and control. Methods The strains of non-O1/non-O139 Vibrio cholerae isolated from sporadic patients and aquatic products from 2015 to 2021 in Zhongshan city were collected. The identification and cluster analysis of the strains were analyzed by matrix assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS), the ctxA virulence gene of strains were detected by real-time fluorescence quantitative PCR, the cluster analysis of the strains was analyzed by pulsed-field gel electrophoresis (PFGE), and the drug resistance of the strains were analyzed by microbroth dilution method. Results From 2015 to 2021, 33 strains of non-O1/non-O139 Vibrio cholerae were isolated from Zhongshan City, including 28 strains from sporadic patients and 5 strains from aquatic products. Through MALDI-TOF-MS identification, 33 strains of non-O1/non-O139 Vibrio cholera can be identified to the level of species, and the identification results were all Vibrio cholerae. Among 33 non-O1/non-O139 Vibrio cholerae strains, 1 strain carried the ctxA virulence gene. The drug-resistant strains accounted for 69.7% (23/33), and the multidrug resistant strains accounted for 18.2% (6/33). A total of 7 kinds of drug resistance spectrum were produced, including 3 kinds of multidrug resistant spectrum, and showed drug resistance to 8 antibiotics, among which the resistance rates to streptomycin, cefazolin and compound sulfamethoxazole were above 30%. The 33 strains of non-O1/non-O139 Vibrio cholerae were divided into 32 PFGE fingerprints with a similarity ranging from 61.7% to 100%. MALDI-TOF-MS cluster analysis divided 33 non-O1/non-O139 Vibrio cholerae strains into two clusters. Conclusions The results of molecular typing of non-O1/non-O139 Vibrio cholerae in Zhongshan City presented diversity, and no significant correlation was found between PFGE and MALDI-TOF-MS cluster analysis. The strains demonstrated various degrees of resistance to certain antibiotics, and there were multidrug-resistant and toxigenic strains. Therefore, it is necessary to alert to the harmfulness of non-O1/non-O139 Vibrio cholerae and enhance monitoring.
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Objective:To set up the rat skeletal muscle L6 cell models of oxygen-glucose deprivation(OGD)in vitro,and to investigate the protective effect of EGF in deep tissue inj ury(DTI)of pressure sores.Methods:The rat skeletal muscle cells in the logarithmic phase were divided into normal control group,OGD group,5 μg·L-1 EGF+OGD group,10 μg·L-1EGF+ OGD group and 20 μg·L-1EGF+ OGD group.The survival rates of skeletal muscle cells in various groups were measured by MTT assay;the cell apoptotic rates in various groups were detected by flow cytometry;the reactive oxygen species(ROS)levels were detected by DCFH-DA;Rhodamine 123 was used to detect the mitochondrial membrane potential;the expressions of Bax and Bcl-2 proteins were determined by Western blotting method.Results:Compared with normal control group,the survival rates of skeletal muscle cells in OGD group after 24 h OGD was significantly decreased(P<0.05);the apoptotic rate was markedly increased(P<0.01);the ROS level was increased(P<0.01);the mitochondrial membrane potential was decreased(P<0.01);the ratio of Bcl-2/Bax was significantly decreased(P<0.01).Compared with OGD group, the survival rates of skeletal muscle cells in different concentrations of EGF groups were increased and the apoptotic rates were decreased,especially in 10 and 20 μg·L-1EGF groups(P<0.05 or P<0.01);the ROS levels in skeletal muscle cells in different concentrations of EGF groups were decreased and the mitochondrial membrane potential were increased,especially in 10 and 20 μg·L-1EGF groups(P<0.05 or P<0.01);the Bcl-2/Bax ratios were significantly decreased in a concentration-dependent manner,especially in 10 and 20 μg·L-1EGF groups (P<0.05 or P<0.01).Conclusion:EGF can improve the skeletal muscle cell injury induced by OGD in a concentration-dependent manner via decreasing the ROS levels and protecting the cell mitochondrial function.
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The new media has brought the hospital propaganda work opportunities and challenges.By analyzing the characteristics of the new media,the study elaborated how to upgrade the hospital propaganda project under the new media environment,and make a new news carrier.The new way should be taken to shape the hospital online image,set up the hospital brand,spread medical information and promote health education.
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Objective To study and analysis the effect of different doses of sodium phosphate solution in bowel preparation for colorectal treatment in children. Methods 100 patients who underwent colonoscopic polypectomy treated in our hospital from February 2015 to September 2016 were randomly divided into group A and group B, each group of 50 patients each. A group was treated with 2 bottles of cleansing liquid heating boiling water, B group were treated with 1 bottles of cleansing liquid heating boiling water. The therapeutic effects of the experimental group and the control group were compared and analyzed. Results The tolerance of group B was 78.0%, significantly better than that of group A(50.0%), and the difference was statistically significant (P<0.05). The incidence of adverse reactions in group B was 24.0%, significantly lower than that in group A (60.0%), and the difference was statistically significant (P<0.05). In group B, cleanliness was common in 1 patients and poor in cleanliness in 3 patients. In the A group, 1 patients had poor cleanliness and 2 patients had general cleanliness. There was no significant difference between the two groups. Conclusion 1 bottles of sodium phosphate in intestinal cleaning colorectal bowel preparation effect is good, low incidence rate of adverse reaction, greatly improve the intestinal cleanliness, with further clinical promotion and application significance.
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Objective To investigate the expression of p21 and p53 in renal interstitial fibrosis rats and the effect of enalapril on it. Methods Sprague-Dawley rats were randomly divided into 3 groups, shame operation rats group, unilateral urethral obstruction and enalapril treatment group. Histological chan-ges were observed by Masson stain. The expression of p21 mRNA and p53 mRNA was detected by RT-PCR. Results With degree of interstitial fibrosis aggravating, the expression of p21 and p53 increasing,p21 and p53 expression of UUO rats at every time point were positive correlative. Enalapril can inhibit the expression of p21 and p53. Concinsion p21 and p53 expression increased in UUO rats renal cortex and enalapril can significantly inhibit its expression, p21 may participate in the pathogenesis of renal tubule-in-terstitial fibrosis through p53 pathway.
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The preparation process and immunogenicity of a novel hepatitis B vaccine containing preS1, preS2 and S epitopes were investigated in this study. A Pichia pastoris stain GS115-SS1S2 harbouring two chimeric HBsAg gene constructs, SS1 and SS2 was cultivated by high-density fermentation. 300-600 mg/L of the expression level was achieved through 48-72 h methanol induction. SSIS2 antigen was extracted and purified by silica adsorption, HIC and SEC to 99% purity from the harvested cells. 82 mg purified antigen could be achieved from one liter of fermentation culture. The immunogenicity of the purified antigen was evaluated in NIH mice. Three groups of female NIH mice, 14-16 g in weight, were injected once intraperitoneally with 2.5, 0.625, 0.156 microg of each of the two vaccines: SS1S2 or a commercially available S vaccine. Part of the mice were bled in 30 days after injection to compare the ED50 of the two vaccines. For the SSIS2 vaccine, the ED50 is 0.46, 0.29 and 0.84 microg respectively for the preS1, preS2 and S antigens. For the S vaccine, the ED50 is 0.99 microg for the S antigen. Another part of the mice were bleed in 7 or 14 days to detect preS1, preS2 and S antibodies. Higher ratios of mice were seroconverted for preS1 and preS2 antibodies as compared to the S antibody in these two time points. These results suggest that the SS1S2 vaccine may be more immunogenic than the conventional S vaccines.
Subject(s)
Animals , Female , Mice , Epitopes , Allergy and Immunology , Hepatitis B Surface Antigens , Allergy and Immunology , Hepatitis B Vaccines , Allergy and Immunology , Pichia , Genetics , Metabolism , Protein Engineering , Protein Precursors , Allergy and Immunology , Recombinant Proteins , Genetics , Allergy and Immunology , Viral Envelope Proteins , Allergy and ImmunologyABSTRACT
At present time, the widely used hepatitis B virus( HBV) vaccines consist of only the small hepatitis B surface antigen expressed in yeast or CHO cells. The frequency of non-responders to these vaccines has increased the demand for a more immunogenic vaccine. Some studies have suggested that the addition of preS region to the vaccine will improve its efficacy. However, the large protein (L) containing the whole preS region can not be effectively expressed in vitro. To overcome this problem, two chimeric contructs, SS1, surface gene containing preS1 region at C-terminus and SS2, surface gene containing preS2 region at C-terminus, were constructed and effectively expressed in our previous studies. Here we further constructed an expression vector containing both SS1 and SS2 expression cassettes by separation and ligation the SS2 cassette to a linearized SS1 expression vector pAO815-SS1. The recombinant vector was transformed into Pichia pastoris GS115 by electroporation. A high-level expression strain (GS115-SS1S2) was established by primary screening for His+ transformants and further analysis for induction products. ELISA results demonstrated that the expressed protein had S, preS1 and preS2 antigenicities simultaneously. Western blotting showed that the product can bind to all of the three antibodies, anti-S, anti-preS1 and anti-preS2. The expression protein was present in the form of particles of 20-35 nm diameter and the yield of recombinant particles reached 300-600 mg/L by fermentation. The SS1 and SS2 polypeptides kept intact in purified particles, suggesting that the stability of preS region has been significantly improved.
Subject(s)
Animals , Cricetinae , CHO Cells , Cricetulus , Epitopes , Genetics , Allergy and Immunology , Hepatitis B Surface Antigens , Genetics , Allergy and Immunology , Hepatitis B Vaccines , Allergy and Immunology , Peptide Fragments , Genetics , Allergy and Immunology , Pichia , Genetics , Protein Precursors , Genetics , Allergy and Immunology , Vaccines, Synthetic , Allergy and ImmunologyABSTRACT
OBJECTIVE@#To investigate the expression of P21 in renal interstitial fibrosis rats and the effect of enalapril on it.@*METHODS@#Sprague Dawley rats were randomly divided into 3 groups: a sham operation group,a unilateral urethral obstruction group, and an enalapril treatment group. The expression of P21 in renal tubular epithelial cells on the process was detected by immunohistochemistry at different time spots (7, 14, 21 d after UUO, sham-surgery or enalapril treatment). The expression of p21 mRNA was detected by reverse transcription-polymerase chain reaction (RT-PCR).@*RESULTS@#Seven days after the surgery, significant differences were found in P21 expression between UUO and SOR renal tubular cells. With degree of interstitial fibrosis aggravating, P21 expression increased. Enalapril can inhibit its expression.@*CONCLUSION@#In the kidney of UUO rats, P21 expression increased and enalapril possessed significant inhibitory effects on the procedure. P21 may participate in the pathogenesis of renal tubule-interstitial fibrosis.
Subject(s)
Animals , Male , Rats , Angiotensin-Converting Enzyme Inhibitors , Pharmacology , Therapeutic Uses , Enalapril , Pharmacology , Therapeutic Uses , Kidney Tubules , Metabolism , Nephrosclerosis , Drug Therapy , Metabolism , Proto-Oncogene Proteins p21(ras) , Genetics , RNA, Messenger , Genetics , Random Allocation , Rats, Sprague-Dawley , Ureteral ObstructionABSTRACT
OBJECTIVE@#To explore the effect of p27 in the renal tubule on the process of renal interstitial fibrosis caused by unilateral ureteral obstruction (UUO) in rats, and to examine the expression changes of p27 after enalapril intervention and to interpret the anti-fibrotic mechanism.@*METHODS@#Ninety rats were randomly divided into the sham-operated group (SOR), UUO group,and UUO+enalapril treatment group [enalapril: 10 mg/(kg.d)]. The rats of each group were respectively sacrificed on 7, 14, 21 days post-operatively. The renal pathological changes were dynamically observed by HE. The expression and dynamic changes of p27 were detected by immunohistochemistry. The level of p27 mRNA were detected by RT-PCR.@*RESULTS@#The expression of p27 in renal tubular epithelial cells and p27 mRNA were strongly positive in the SOR group. With degree of interstitial fibrosis aggravating, the expression of p27 mRNA was gradually reducing. Enalapril could improve the expression of p27 on the 14th and 21st days after the UUO.@*CONCLUSION@#(1) This study supports a causative role of p27 in the formation of fibrosis of renal mesenchyme in rats with UUO. (2) The anti-fibrotic mechanism of enalapril is partly the improvement of p27 expression.
Subject(s)
Animals , Female , Rats , Angiotensin-Converting Enzyme Inhibitors , Pharmacology , Therapeutic Uses , Cyclin-Dependent Kinase Inhibitor p27 , Genetics , Enalapril , Pharmacology , Therapeutic Uses , Kidney Tubules , Metabolism , Nephrosclerosis , Drug Therapy , Metabolism , RNA, Messenger , Genetics , Random Allocation , Rats, Sprague-Dawley , Ureteral ObstructionABSTRACT
Allogeneic hematopoietic stem cell transplantation is curative therapy for many malignant hematologic and genetic disorders. The pathways by which T cell are regenerated in vivo are of central importance for host immune competence after the transplantation. Thymic-dependent and thymic-independent pathways of T-cell regeneration are primarily determined by thymus function, and that thymic-independent pathways are generally inadequate for restoration of host immunocompetence. T cell regeneration of adult recipients is largely through thymic-independent pathways due to reduced thymic regeneration capacity, resulting in quantitative defficiencies in T-cell number and severe restriction in the diversity of the regenerated T-cell receptor (TCR) repertoire. New strategies to enhance thymic function in man after BMT would hold great therapeutic potential. Activation of donor T cells is the cause of GVHD, the induction of anergy can inactivate specific sets of alloreactive T cells in the donor stem cells, which can prevent GVHD without inhibiting the entire T-cell repertoire.
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To study the reconstituion of the T-cell populations after allogeneic hematopoietic stem cell transplantation, the peripheral blood samples obtained at different time points post-transplantation in 21 patients were analyzed using CD5-PE and CD3-FITC with flow cytometry. During the first 3 months after transplantation, CD3(-) CD5(+) T cell subsets, representing early thymocytes, remained below normal level, whereas CD3(+) CD5(+) T cell subsets, representing mature T cells, regenerated to normal level. In 9 patients who developed acute graft-versus-host disease (GVHD), the percentage of CD3(+) CD5(+) T cell subsets was significantly higher than that in patients who did not develop acute GVHD (P < 0.05). These results demonstrated that the early recovery of T cells is mainly due to the expansion of mature T cell populations, and the over-expansion of mature T (CD3(+) CD5(+)) cells is responsible for the development of acute GVHD.