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ObjectiveTo investigate the bactericidal effect of loaded multifunctional povidoneiodine-nanometer selenium （PVP-I@Se） disinfectant on Staphylococcus aureus （SA） and methicillin-resistant Staphylococcus aureus （MRSA）， and to provide an experimental basis for the reduction of surgical site infection （SSI）. MethodsThe control group was the povidone iodine （PVP-I） group with different concentrations of iodine （50， 75， 100， 200 and 400 μg/mL）. The PVP-I@Se group （experimental group） was the PVP-I group further supplemented with 2 μg/mL Selenium nanoparticles （SeNPs）. Then we compared the bactericidal effect of the two groups of disinfectant solutions on SA and MRSA by examining the minimum inhibitory concentration （MIC）， minimum bactericidal concentration （MBC）， the shortest sterilization time at a concentration of 50 μg/mL iodine and the inhibition zone diameters at concentrations of 200 μg/mL and 400 μg/mL iodine. ResultsMIC values of PVP-I against SA and MRSA were both 79.17 μg/mL， and those of PVP-I@Se were 54.17 and 70.83 μg/mL， respectively. MBC values of PVP-I against SA and MRSA were 129.17 and 150.00 μg/mL， respectively， and those of PVP-I@Se were 70.83 and 87.50 μg/mL， respectively. At a concentration of 50 μg/mL iodine， the shortest sterilization time of PVP-I for SA and MRSA was 130 s and 140 s， respectively， and that of PVP-I@Se was 65 s and 75 s， respectively. At a concentration of 200 μg/ml iodine， the inhibition zone diameters of PVP-I for SA and MRSA were 7.67 mm and 8.33 mm， and those of PVP-I@Se were both 9.50 mm. At a concentration of 400 μg/mL iodine， the inhibition zone diameters of PVP-I for SA and MRSA were 9.00 mm and 9.33 mm， and those of PVP-I@Se were 11.67 mm and 12.00 mm， respectively. ConclusionsPVP-I with different concentrations of 50， 75， 100， 200 and 400 μg/mL iodine supplemented with 2 μg/mL SeNPs have better and faster bactericidal effect on SA and MRSA. When combined with SeNPs， PVP-I can enhance the bactericidal activity against SA and MRSA， but with better sensitizing effect on SA than MRSA and higher demand of iodine concentration （400 μg/mL） for sensitizing effect on MRSA. This study provides a theoretical basis for selecting optimal concentration and action time of the disinfectant， thus reducing SSI.
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【Objective】 To investigate the variation of hematological parameters in male plateletpheresis donors. 【Methods】 A total of 194 male plateletpheresis donors from Fujian Blood Center were divided into two groups according to the frequency of blood donation: Group 1 (n=107), with the number of plateletpheresis donation less than or equal to 12 per year; Group 2 (n=87), with the number of plateletpheresis donation more than 12 per year. Serum ferritin (SF) and related iron metabolism indexes, red blood cell count (RBC), hemoglobin (Hb), hematocrit(Hct), platelet count (Plt) and other blood routine indexes, as well as percentage of reticulocyte counts (RET%), immature reticulocyte fraction(IRF) and other reticulocyte indexes were measured before blood donation and analyzed by statistical methods. 【Results】 Compared with Group 2, the RBC, Hb, Hct, SF in Group 1 were significantly higher, while Plt, RET%and IRF were significantly lower(P<0.05), and the probability of ferritin decrease in Group 1 was lower, with significant difference(P<0.05). 【Conclusion】 As the number of donation increased, male plateletpheresis donors were prone to iron deficiency, and the bone marrow hematopoiesis were obviously enhanced. We should be more concerned about male plateletpheresis donors who donated more than 12 times per year, further more, SF monitoring should be conducted and the blood donation interval should be appropriately extended.
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Objective:The study was planned to evaluate the clinical utility of microcytic anemia factor (MAF) and low hemoglobin density (LHD%) in the screening of iron deficiency in blood donors.Methods:A total of 400 blood donors, 200 male and 200 female, were randomly admitted to Fujian Blood Center from January1, 2022 to February 28, 2022 by the way of stratified sampling. According to the fourth edition of Diagnostic and therapeutic criteria for hematological diseases, the patients were divided into three groups: normal group (N=299), iron depletion group (ID, n=54) and iron deficient erythropoiesis group (IDE, n=47), Blood routine indexes including hemoglobin (HGB), mean corpuscular volume hemoglobin (MCV), mean corpuscular hemoglobin content (MCH), mean corpuscular hemoglobin concentration (MCHC) and iron metabolism indexes including serum ferritin (SF), serum iron (SI), total iron binding capacity (TIBC), transferrin saturation (TS) and unsaturated iron binding capacity (UIBC) were measured, MAF and LHD% were calculated by formula.One-way ANOVA or Kruskal-Wallis H tests were used to analyze the differences among three groups. Spearman correlation analysis was used to analyze the correlation between MAF and LHD% and iron metabolism indexes.The Receiver Operating Characteristic (ROC) curve was used to evaluated the diagnostic value of MCH, MCHC, MAF and LHD% for iron deficiency in blood donors. Results:MAF in ID group which was 11.81±0.81 were higher than the IDE group which was 10.69±0.95 and lower than the healthy group which was 13.17±1.24, the total difference among the three groups was statistically significant ( F=110.784, P<0.001), the difference between two groups was statistically significant ( P<0.01); LHD% in ID group which was 2.61 (1.87, 3.91)% were lower than the IDE group which was5.60(2.99, 8.02)% and higher than the healthy group which was1.74 (1.22, 2.73)%, the total difference among the three groups was statistically significant ( H=62.166, P<0.001), the difference between two groups was statistically significant ( P<0.01).In 101 iron deficiency blood donors, Spearman correlation analysis showed that MAF was positively correlated with SF, SI and TS ( r=0.426, P<0.01; r=0.547, P<0.01; r=0.566, P<0.01);contrarily, LHD% was negatively correlated with SF, SI and TS ( r=-0.397, P<0.01; r=-0.400, P<0.01; r=-0.479, P<0.01).The areas under the ROC curve of MCH, MCHC, MAF and LHD% diagnostic ID were 0.745, 0.646, 0.819 and 0.646, respectively;the cut-off value of MAF was 12.56, with a sensitivity of 67.90% and a specificity of 83.30%.While the areas under the ROC curve of MCH, MCHC, MAF and LHD% diagnostic IDE were 0.901, 0.834, 0.941 and 0.834, respectively; the cut-off value of MAF was 11.73, with a sensitivity of 87.60% and a specificity of 87.20%. Conclusions:MAF performed a high diagnostic value of iron deficiency, especially IDE, and can be used as a marker in the diagnosis of iron deficiency in blood donors.
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Objective To explore the effects of early enteral nutrition (EN) combined with probiotics on intestinal flora and immune function in patients with severe ischemic stroke. Methods Sixty-nine severe ischemic stroke patients were admitted and continuously enrolled in Taizhou First People's Hospital from June 2017 to June 2018, and they were randomly divided into an EN combined with probiotics group (35 cases) and a simple EN group (34 cases). Early EN support was given to both groups and probiotics (Live Combined Bifidobacterium, Lactobacillus and Enterococcus capsules) was added to the EN combined with probiotics group, 0.42 g each time, 3 times a day for 14 days. The changes of serum inflammatory markers [hypersensitive C-reactive protein (hs-CRP), procalcitonin (PCT), tumor necrosis factor-α (TNF-α), interleukin-10 (IL-10)], intestinal flora (Bifidobacterium, Lactobacillus, Clostridium, Enterobacter, Enterococcus, Bacteroides), intestinal mucosal barrier [endotoxin, D-lactic acid, diamine oxidase (DAO), intestinal fatty acid binding protein (I-FABP) ], and immune indexes [immunoglobulins (IgA, IgG, IgM), human leukocyte DR antigen (HLA-DR)] were observed in two groups of patients after treatment. Results With the prolongation of time, Bifidobacteria, Lactobacilli, HLA-DR and IgA, IgG, IgM after EN in both groups all decreased first and then had a tendency of increase, all reaching the lowest value on the EN 3rd day and then gradually elevated arriving at the peak value on the EN 14th day, and the levels in EN combined with probiotics group were significantly higher than those in the simple EN group [Bifidobacterium (×107 cfu/g): 8.31±1.49 vs. 7.49±1.32, Lactobacillus (×107 cfu/g): 8.04±1.45 vs. 7.19 ±1.37, HLA-DR: (67.22±9.11)% vs. (61.21±9.69)%, IgA (mg/L): 170.34±40.13 vs. 149.54±38.76, IgG (g/L):4.88±0.88 vs. 4.31±0.86, IgM (mg/L): 879.47±100.82 vs. 821.52±97.75, all P < 0.05]. With the prolongation of time, the Clostridium, Enterobacter, Enterococcus, Bacteroides, hs-CRP, PCT, TNF-α, endotoxin, D-lactic acid, DAO, I-FABP after En in both groups all increased first and then had a tendency of decrease, reaching the highest level on the EN 3rd day, then gradually decreased arriving at the valley value on the EN 14th day, and the levels in the EN combined with probiotics group were significantly lower than those in the simple EN group [Clostridium (×107 cfu/g): 5.23±0.87 vs. 5.79±0.91, Enterobacter (×107 cfu/g): 7.45±1.21 vs. 8.62±1.32, Enterococcus (×107 cfu/g): 7.32±1.05 vs. 8.12±1.23, Bacteroides (×107 cfu/g): 9.16±1.35 vs. 9.87±1.42, hs-CRP (mg/L): 18.45±12.98 vs. 25.47±15.55, PCT (ng/L): 3.24±1.21 vs. 4.18±1.32, TNF-α (ng/L): 9.43±8.69 vs. 13.59±9.45, IL-10 (μg/L): 39.45±10.72 vs. 48.52±11.42, endotoxin (U/L): 6.74±2.12 vs. 9.21±3.28, D-lactic acid (mg/L): 98.74±20.74 vs. 114.78±19.89, DAO (mg/L): 21.45±8.49 vs. 29.47±9.41, I-FABP (ng/L): 1.4±0.2 vs. 1.5±0.2, all P < 0.05]. Conclusion Early EN combined with probiotics can effectively regulate the intestinal flora and intestinal mucosal barrier function, reduce the level of inflammatory response and enhance the body immunity in patients with severe ischemic stroke.
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Background and purpose:Cancer of unknown primary (CUP) represents approximately 5%~10%of malignant neoplasms. For CUP patients, identiifcation of tumor origin allows for more speciifc therapeutic regimens and improves outcomes.Methods:By retrieving the gene expression data from ArrayExpress and Gene Expression Omnibus data repositories, we established a comprehensive gene expression database of 5 800 tumor samples encom-passing 22 main tumor types. The support vector machine-recursive feature elimination algorithm was used for feature selection and classiifcation modelling. We further optimized the RNA isolation and real-time quantitative polymerase chain reaction (RTQ-PCR) methods for candidate gene expression proifling and applied the RTQ-PCR assays to a set of formalin-fixed, paraffin-embedded tumor samples.Results:Based on the pan-cancer transcriptome database, we identiifed a list of 96-tumor speciifc genes, including common tumor markers, such as cadherin 1 (CDH1), kallikrein-re-lated peptidase 3 (KLK3), and epidermal growth factor receptor (EGFR). Furthermore, we successfully translated the microarray-based gene expression signature to the RTQ-PCR assays, which allowed an overall success rate of 88.4% (95%CI: 83.2%-92.4%) in classifying 22 different tumor types of 206 formalin-fixed, paraffin-embedded samples. Conclusion:The 96-gene RTQ-PCR assay represents a useful tool for accurately identifying tumor origins. The assay uses RTQ-PCR and routine formalin-ifxed, paraffn-embedded samples, making it suitable for rapid clinical adoption.
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OBJECTIVE:To establish a method for the limit detection of aconitine in Wuqi shujin tongluo tablet. METHODS:TLC was conducted to identify the aconitine;TLC plate was silica gel G plate,developing solvent was toluene-ethyl acetate-chloro-form-acetone-ammonia(20:18:3:6:1,V/V/V/V/V),chromogenic agent was bismuth potassium iodide test solution and sodium ni-trite ethanol test solution;and durability investigation and detection limit detection were used to optimize the TLC plate,tempera-ture and humidity. RESULTS:TLC showed the aconitine had clear spots and negative control without interference. The durability was good;detection limit was 0.9 μg;available TLC plate was Merck HPTLC prefabricated plate, silica gel G TLC plate setf-made silica gel G TLC plate with adhesive of sodium carboxymethycellulose;temperature was 5-16 ℃ and humidity was 32%-72%. CONCLUSIONS:The method is simple and reproducibility,and can be used for the limit detection of aconitine in Wuqi shujin tongluo tablet.
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ObjectiveTo study the effect of a recombinant plasmid encoding mouse interleukin 2 (mlL-2) on the immunogenicity of DNA vaccine against Chlamydia trachomatis(Ct) serovar E.Methods BALB/c mice were divided into 4 groups to be intramuscularly inoculated with blank plasmid(negative control group),DNA vaccine against Ct serovar E(DNA vaccine group),DNA vaccine against Ct serovar E and a recombinant plasmid containing mIL-2(combination group),and inactivated Ct serovar E elementary bodies (positive control group),respectively.The immunological effects were evaluated by posterior foot pad thickness,proliferation level of spleen lymphocytes,serum level of IL-4 and interferon (IFN)-γ in mice,and the capability to clear Ct genital tract infection.ResultsThe proliferation index of spleen lymphocytes in the combination group and positive control group was similar(3.64 ± 0.41 vs.3.77 ± 0.34),but was significantly different from that in the blank control group and DNA vaccine group (1.37 ± 0.21 and 2.52 ± 0.30).The serum level of IL-4 was(38.49 ± 12.24) pg/ml in the positive control group,significantly higher than in the negative control group,DNA vaccine group and combination group ((25.37 ± 18.93),(24.75 ± 8.49),(21.74 ± 6.43) pg/ml,respectively).With respect to the serum level of IFN-γ,the combination group and positive control group were similar ((1923.3 ± 518.1) pg/ml vs.(2712.5 ± 887.2) pg/ml),but were significantly different from the negative control group and vaccine group((310.8 ± 160.7) pg/ml and(601.3 ± 357.9) pg/ml).Six days after Ct challenge,the exfoliated cells from genital tract were positive for Ct culture in the negative control group,but negative in the other 3 groups.ConclusionIL-2 genetic adjuvant can enhance the immune response,especially Th1 type response,induced by the DNA vaccine against Ct serovar E.
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Objective To investigate the character and location of a novel gene R049 and its expressed protein in uropathogenic Escherichia coli(UPEC) strain 132 isolated in China. MethodsThe chromosome library of UPEC132 was constructed by a shotgun strategy and the sequence analysis was carried out by a high-throughput pyrophosphate sequencing. Sequence reads were assembled with the Newbler program.The characters of R049-associated specific fragment were analyzed using the bioinformatics methods. Outer and inner membrane proteins of UPEC132 were extracted and then detected by SDS-PAGE and Western blot analysis together with the whole-cell lysates. ResultsThe 169 022 bp contig containing gene R049 was obtained and its sequence was very similar to the chromosome associated sequence of UPEC strain 536. It showed that a 20 773 bp fragment including R049 replaced the pathogenicity island PAI Ⅲ536 of UPEC536 in above 169 022 bp contig. The fragment had a lower GC content (46.97％) and 16 bp direct repeats in two ends. Significantly it also was adjacented to thrW tRNA, insertion element and genes coding integrase. Thus the 20 773 bp fragment was named R049 genome island(R049-GI). There were 25 ORFs in R049-GI, and gene R049 was located in the thirteenth ORF. The results of SDS-PAGE and Western blot revealed gene R049 encoded an outer membrane protein in the size of 47.0× 103. ConclusionGene R049, encoding an outer membrane protein, was a component part of the genome island in UPEC 132 chromosome acquired by horizontal gene transfer.
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Objective To study cellular immune responses induced by DNA vaccine against Chlamydia trachomatis (Ct) serotype E. Methods BALB/c mice were divided into three groups to be intramuscularly immunized by blank plasmid (negative control group), DNA vaccine against Ct serotype E (vaccine group), and inactivated Ct elementary body (positive control group), respectively. Two weeks after the last immunization,delayed-type hypersensitivity (DTH) response was evaluated; MTT assay was performed to detect the proliferation of spleen lymphocytes, ELISA to measure the serum level of interferon-γin mice. Some immunized mice underwent a genital challenge with Ct elementary body followed by isolation of Ct from exfoliated epithelial cells in genital tract and pathological examination of cervical tissue from the challenged mice. Results Compared to negative control group, vaccine group and positive control group experienced a stronger DTH response.The lymphocyte stimulating index and serum level of IFN-γwere highest in the positive control group (3.81 ±0.30, 2891.7 ± 1048.8 μg/L), followed by vaccine group (2.35 ± 0.25, 593.3 ± 342.6 μg/L) and negative control group (1.48 ± 0.15, 309.2 ± 157.9 μg/L), and significant difference was observed between the three groups (P ＜ 0.05 or 0.01 ). After Ct challenge, Ct was isolated from exfoliated epithelial cells and cervical tissue was damaged in the negative control group, while in the other two groups, Ct was undetected and genital tract tissue was intact. Conclusions The DNA vaccine against Ct serotype E could induce Ct-specific cellular immune responses to some extent, and offer a protection against vaginal challenge with Ct.
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Objective To investigate the interaction between uropathogenic Escherichia coli (UPEC) and host uroepithelial cells, define the role uroepithelial cells play in initiating and modulating the host response to infection with UPEC strain. Methods The human bladder transitional epithelial EJ cells were evaluated for their capacities to allow the adherence and invasion by UPEC132, a clinical strain isolated from Tianjin, China, and a cDNA microarray for 22 000 human genes was used to identify the gene expression differences between EJ cells infected with UPEC132 and uninfected EJ cells. Results Microscope observation showed that UPEC132 could adhere to EJ cells with the adherence rate of (73.20 ± 5.26)%. And visualization by confocal microscope revealed that this microorganism could be seen within the cells. EJ cells infected with UPEC132 changed mRNA expression of a total of 29 genes, including 28 genes up-regulated and 1 gene down-regulated. Of these, regulators of growth and proliferation, cytokines, and modulators of apoptotic responses were the most prominent. Conclusion The gene expression profiling of EJ cells is affected by the infection of UPEC strain. The differentially expressed genes may contribute to further investigate the interaction of UPEC and uroepithelial cells.
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Objective To investigate the clinical manifestation of hMPV in infants and young chil-dren presented with acute respiratory tract infection and to identify the molecular character. Methods Na-sopharyngeal aspirates were taken from 310 hospitalized pediatric patients from February to May in 2006, March to April in 2008, and September 2008 to February 2009, and the N gene fragments of hMPV were de-tected by nested PCR amplification. Phylogenetic analysis of 17 strains hMPV N genes was performed. The clinical materials of patients were collected and analyzed. All hMPV-positive samples were examined by multi-PCR for other respiratory viruses. Results Of 310 pediatric patients, 20 (6.5%) were positive for hMPV. The median age of hMPV infected children was 15.0 months(from 16 days to 9 years old), 90% (18/20)of the cases were under 2 years, and 60% were male. Phylogenetic analysis of 17 N gene fragments showed that 11 hMPV strains were A2b subtype. 20 hMPV-positive children were subjected to pneumonia, accounting for 7.1% (20/282) among all pneumonia subjects in this study. The common clinical manifesta-tions of hMPV infected patients were cough, wheezing, shortness of breath and fever. 35% (7/20) needed intensive care, 15% (3/20) were given oxygen therapy. The median length of hospital stay was (11.9 ±4.8) d. No significant seasonal distribution of hMPV was displayed. Two patients were coinfected with ade-novirus and rhinovirus respectively. Conclusion hMPV was an important respiratory pathogen in young children subjected to pneumonia in Tianjin. Three subtypes(A2a/A2b, B1, B2) were prevalent in Tianjin, and A2b was the predominant subtype. No significant difference of clinical characters was observed between A and B type hMPV infected patients.
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Objective To investigate expression of polysaccharides biosynthetic genes in biofilm formation in Pseudomonas aeruginosa PAO1. Methods Real-time RT-PCR was used to quantify mRNA to analyze their mRNA level during planktonic growth and biofilm cells. Results Analysis of the relative ex-pression levels indicated that the level of transcripts of planktonic pslA, algD, pelA was significantly lower than that of biofilms sessile growth in all other form bacteria. The levels of transcripts of pslA, algD, pelA were the highest at the first day of biofilms development. Conclusion From the observations in the present study, transcripts of the pslA, algD, pelA involved biofilm development.
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Objective To discuss the clinical value and nursing of the three-endoscope in the treatment of eholedoeholithiasis. Methods 45 eases of choledocholithiasis patients who were treated with LCDE (three-endoscope) were named as the research group.56 patients who received traditional open ab-dominal surgery were set as the control group. The average hospitalization time and satisfaction degree with nursing were compared, t test and χ2 test were adopted. Results The average hospitalization time was shorter and satisfaction degree with nursing was higher in the research group than those in the control group. Conclusions The treatment of choledochohthiasis with three-endoscope is safe and feasible, es-pecially when combined with antibiotics lavage and stone dissolution through naso-biliary duct.The opera-tion can widen the surgical indication,reduce the risk of surgery with little damage,clear stones completely, reduce postoperative complicatioas,make patients recover faster, shorten the hospital stay and achieve the same or better treatment results when compared to the traditional open abdominal surgery.
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Objective To clone and express the new gene R049 of uropathogenic Escherichia coli 132.and to investigate the immunopmtective effects of the R049 recombinant protein on mice.Methods The pmkaryotic expression system of gene R049 was constructed by directed cloning.Thereafter,the R049 recombinant protein Was expressed and purified by Ni affinity chromatography.Polyclonal antibody was pre-pared by immunizing BALB/c mice with R049 recombinant protein.The R049 recombinant protein and whole bacterial proteins of UPEC132 were analyzed by SDS-PAGE and Western blot.BALB/c mice were im-munized with R049 recombinant protein before challenged by UPECl32 through urinary tract.Then the differences of urine and renal colony counts between immunization group and control group were compared.Results The recombinant strain E coli BL21(DE3)/pET32a-R049 ORF was constructed successfully,and the relative molecular mass of the R049 recombinant protein was 66.9×103 and its purity was up to 95% af-ter purification.The titer of polyclonal antibody wag≥1:102 400 analyzed by indirect ELISA.Both of the R049 recombinant protein and whole bacterial proteins of UPECl 32 were confirmed to show specffic reactions on the antiserunl throughh Western blot.The animal experiments showed the urine and renal colony counts of immunization group were significantly lower than that of the control group(P<0.01,P<0.05).Conclu-sion The new gene R049 of uropathogenic E.coli 132 had immunopmtective effects on mice and the defini-tive mechanism would be needed to further study.
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Objective To study the distribution of the specific fragment R049 of uropathogenic E. coli(UPEC) 132 in UPEC and fecal E. coli strains. Methods The specific fragment R049 was amplified by PCR from 20 UPEC strains and 40 fecal E. coli strains, and 5 genes encoding virulence factors (papC, fimH, hly, aer, cnf1) were detected from fragment R049 positive strains. Pulse field gel electronphoresis (PFGE) was applied for isolating the Xba Ⅰ restriction fragments of the genomes of fragment R049 positive strains, and then Southern blot was applied for analyzing the distribution features of the positive hybridization bands by digoxin-labeled R049 ORF probe. Results The specific fragment R049 was amplified from 8 of 20 UPEC strains (40%) and 3 of 40 fecal E. coli strains (7.5%), and statistics analysis showed significant difference (P<0.01). The specific fragment 11049 was closely related with 5 virulence factors of UPEC in the fragment R049 positive strains. Southern blot showed the sizes of positive bands were 150 kb, 15 kb and 240 kb in 3 fecal E. coli strains, 350 kb in 6 of 8 UPEC strains, and 280 kb and 25 kb in the rest two UPEC strains. Conclusion The specific fragment R049 of UPEC132 possessed the feature of clustering distribu-tion in domestic isolated UPEC strains.
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Objective To study proteome variation between uropathogenic E. coil (UPEC)132, UPEC J96 and non-uropathogenic E. coli K-12 MG1655. Methods Two-dimensional gel electrophoresis(2-DE) was applied to compare the differential expression proteins between UPEC 132, UPEC J96 and non-uro-pathogenic E.coli K-12 MG1655. The differential expression proteins were digested in gel by enzyme. The mass of generated peptides were measured by matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS). The data obtained from peptide mass fingerprinting (PMF) were re-searched using the internet available database. Results The number of protein spots recognized from UPEC 132 was 466±11, significantly more than that of E. coli K-12 MG1655 (338±15) and UPEC J96 (382±12); there were 298 protein spots shared by the three E.coli strains, 56 protein spots shared by two UPEC strains, and 89 protein spots characterized by UPEC 132. Twenty-two differential expression or significantly increased expression protein spots, involved in virulence factors, metabolism and transportation, regulation of protein synthesis, biological oxidation and unknown functions, were successfully identified by MALDI-TOF-MS. Condusion The proteome from UPEC 132 and non-uropathogenic E. coli K-12 MG1655, or UPEC 132 and UPEC J96 was differentially expressed. It will provide important information on the pathogen-esis of UPEC 132.
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Objective To establish RT-PCR-RFLP method for studying the genotype of wild mea-sles virus strains isolated from Tianjin area from 2002 to 2008. Methods Isolations of measles virus were carried out by tissue culture method from urine and throat swab specimens collected from suspected cases. RNA were extracted from the virus specimens. The 594 bp fragment of C terminal of the N (nucleoprotein) gene was amplified by one-step RT-PCR, then the PCR products were digested with Bcn I , separated on agarose gel electrophoresis and then analyzed by the method of RFLP (restriction fragment length polymor-phism). In addition, above results were compared with DNA sequencing. Phylogenetic tree was plotted based on the results for the genetic relationship and distance analysis. Results Sixty-nine measles virus strains were isolated from 189 specimens from 2002 to 2008, of which the C terminals of N gene were all de-tected positive. Among the 69 strains of measles virus isolates, 98.55% (68/69) belonged to Hla sub-geno-type which was the predominant sub-genotype, and only one strain (1.45%) belonged to H1b sub-genotype by RFLP analysis which was in accordance with the results by DNA sequencing method. Phylogenetic tree analysis indicated the H1a sub-genotype measles virus strains should be further divided into 2 clades, and the variation fluctuated between 0.2% and 3.8%. There were transmission chains caused by different virus strains co-cireulation. Conclusion A genotype, H1a and H1b sub-genotype can be identified by RT-PCR-RFLP assay specically based on the restriction enzyme Bcn I .The RT-PCR-RFLP assay can be a rapid, simple, accurate and efficient method for large-scale surveillance of measles virus strains in China.
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Objective To construct the multi-epitope antigen(mea) gene of G2 glycoprotein of Hantavirus SEO type L99 strain.Methods The B cell epitopes was chosen and connected by three-peptide GPG after the amino acid sequence of G2 protein was analyzed and predicted by bioinformatical soft wares.The corresponding gene mea was constructed by overlap PCR,and cloned into the prokaryotic expression plasmid pET32a(+).Results The mea gene was successfully constructed by the five epitopes being chosen.The recombinant plasmid pET32a-mea was acquired by directional cloning.Conclusion The mea and its expression system E.coli BL21/pET32a-mea were constructed for the first time.The foundation was laid for the expression of the mea and its immunological applications.
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Objective To screen the high-efficiency predominant bacteria which can decompound 2-mercaptobenzothiazole, the accelerant for producing latex, in the organic wastewater. Methods Sampling from manufacturing environment, we got the predominant bacteria by primary screening, isolating and functional tests, and performed simulated test ground decompounding tests by using all bacteria. The enrichment of the predominant bacteria was followed by screen and identification to select the high-efficiency bacteria. Results 75 strains of predominant bacteria were obtained by primary screening. The simulated decompounding tests were performed after the mixed bacteria were tamed. The ratio of elimination for chemical oxygen demand (COD) was about 60.8%-97.7%, and the average was 77.2%. The predominant bacteria adhered to the surface of the active carbon, the carrier, and formed the biological film. Through screening and identification the Bacillus cereus showed to be predominant (90%). Conclusion The technology of high-efficiency predominant bacteria can be used for decompounding 2-mercaptobenzothiazole in the organic wastewater.
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The bioterrorism-related anthrax, the general situation of anthrax prevalence in China and the latest advances in laboratory diagnosis of Bacillus anthracis were reviewed. The control and prevention measures on anthrax were also introduced and the measures responding to the possible bioterrorism-attack were proposed in this paper.