0.05) after the training,while for the students of control group,the levels of IAP and Npt increased significantly after the training(P

Objective: To isolate and screen active marine microorganisms from the East China Sea and to identify the phenotypes of the isolated strains. Methods: Thirty strains of bacteria isolated from the East China Sea were selected randomly, and the active strains were screened out by the anti-Escherichia coli model. The active strains were subjected to physiological and biochemical characteristics, salt-aggregation test, and 16S rDNA sequence analysis. Neighbor-joining trees were constructed by comparing the results of 16S rDNA sequences with sequences described in the BLAST server of the National Center for Biotechnology Information (NCBI); the strains were subsequently identified to genus level. Results: Strains F81612 and F201721 were screened out with the optimum salinities of 10% and 7.5%, respectively. Their morphology and biochemical characteristics were similar to those of Bacillus sp.. 16S rDNA sequence analysis showed that sequences of F81612 had a higher similarity to those of Bacillus subtilis; F201721 was similar to Bacillus amylolique faciens. Conclusion: Two Macrolactin S-producing strains have been screened out by the anti-Escherichia coli model, and they are identified as moderate halophilic Bacillus sp.

Objective: To exploit marine microorganisms and study their secondary metabolites for new drugs. Methods: An antibacterial model was used to screen for active strains. The ethyl acetate (EtOAc) extract was separated by silica chromatography, gel filtration chromatography, and high-performance liquid chromatography. The structures of the compounds were elucidated by 1 HNMR, 13CNMR and MS technologies; Kirby-Bauer Disc Diffusion method and MTT method were employed to detect the biological activities of the separated compounds. Results: Eleven compounds were separated and identified as macrolactin A (1), 3-Hydroxyl acetyl-indole (2), 3-indolethanol (3), cyclo-(Try-Pro) (4), cyclo-(Ile-Try) (5), cyclo-(Leu-Pro) (6), cyclo-(Leu-Val) (7), cyclo-(Ile-Pro) (8), cyclo-(Phe-Val) (9), N- phenethylacetamide (10), P-hydroxy benzaldehyde (PHB) (11). Conclusion: Compound 1 shows strong inhibitory activities against Pyricularia oryzae, Escherichia coli and Staphylococcus aureus (with MIC values being 3.6, 0.45 and 6.3 μg/ml, respectively), and tumor cell lines HeLa and HepG2 (with the IC50 values being 2.0 and 1.8 μg/ml, respectively).

ObjectiveTo probe into the maternal care utilization by minority women, for the purpose of policy recommendations on better maternal care in minority areas. MethodsA combination of stratified random sampling and typical sampling was made on 445 married women of reproductive age in six counties in Yunnan, Guizhou, Qinghai and Tibet provinces, a field survey on their utilization of maternal care services. ResultsTheir average prenatal detection rate is 78.24％, a level lower than the national rural average of 93.7％ and grade-4 rural average of 81.2％ in 2008; their post partook rate is 30.7％, lower than the national rural average of 54.3％ and grade-4 rural average of 58.9％ in the same period; their average coverage rate is 52.18％, a level lower than the national rural average of 87.1％and grade-4 rural average of 64.3％ in 2008. ConclusionThe maternal care utilization is found to be low for women in minority areas. Effective solutions are expected for payment of indirect expenditure of hospital delivery; better health education for enhancing health knowledge and health awareness of minority women; effective incentive mechanism for village doctors, consolidating the base of the three-level healthcare network.

Objective To investigate the change of splenocyte subsets in Balb/c mice immunized with transgenic alfalfa(Medicago sativa)containing Eg95-EgA31 fusion gene of Echinococcus granulosus(Eg) and challenged with Eg protoscoleces.Methods Leaf protein was extracted from transgenic alfalfa containing Eg95-EgA31 fusion gene by heat-coagulation method,and concentration of 20 g/L was used in the study.Meanwhile,leaf protein extracted from the transgenic alfalfa containing blank vector(pBI121)and the normal alfalfa was served as control.Thirty-two female Balb/c mice were randomly divided into 4 groups,8 mice in each group.Oral group was immunized with the leaf protein containing Eg95-EgA31 fusion antigen intragastrically(100μl per mouse);intranasal group was immunized with the leaf protein containing Eg95-EgA31 fusion antigen intranasally(10 μl per mouse);blank vector group was vaccinated intranasally with 10μl leaf protein with blank vector(pBI121);and normal control group was given 100μl normal leaf protein intragastrically.All mice in the above mentioned groups were immunized every 3 days for 2 months.Then,the mice were challenged intraperitoneally with Eg protoscoleces(50 protoscoleces per mouse)8 weeks after last vaccination and sacrified 24 weeks pest infection to separate the splenocytes.Flow cytometry was used to measure the percentages of CD4+ and CD8+ T ceils subsets.Resuits Compared with the normal control group(0.166±0.018,0.083±0.006,2.019 ±0.369),the percentages of CD4+(0.286±0.009)and CD8+(0.102±0.004)T cell subsets and the ratio of CD4+/CD8+(2.814±0.014)in oral group increased significantly (P<0.01 or<0.05).The percentage of CD4+ subset(0.269±0.016)and the ratio of CD4+/CD8+(2.955±0.986) in intranasal group was significantly higher than that ofthe normal control group(all P<0.01).The percentage of CD4+ subset in oral group was significantly higher than that of the intranasal group(P<0.05).No significant difference was found in the percentages of CD4+ and CD8+ T cell subsets and the ratio of CD4+/CD8+ between the blank vector group(0.169±0.018,0.093±0.019,1.852±0.188)and the normal control group(all P>0.05).Conclusions CD4+ T cell may play an important role in the protection induced by transgenic alfalfa vaccine against the challenge of Eg protoscoleces.Intragastrical immunization may be a good route.

Objective To investigate the dynamic changes of splenocyte cytokines in mice immunized with the transgenic alfalfa containing Eg95-EgA31 fusion gene of Echinococcus granulosus (Eg). Methods Eighty-eight Balb/c mice were divided into 2 groups randomly according to body weights,and immunized orally or intranasally with 100μl or 10μl extracted leaf protein from the transgenic alfalfa(20 g/L) respectively once per 3 days for 2 months. Four mice randomized from each group were killed to get splenocyte on week 0(control),2,4,6,8,10,12,14,16,18 and 20 after the last immunization. The splenocyte were cultured in medium for 48 hours with EgAg or concanavalin A (ConA) stimulation to induce the interleukin (IL)-12,interferon γ(IFN-γ) and IL-10,and cultured for 72 hours with EgAg or lipopolysaccharide (LPS) stimulus to induce the tumor necrosis factor α (TNF-α). Then the supernatant was collected to measure the level of IL-12,IFN-γ,TNF-α and IL-10 by ELISA. Results In the oral immunization group,the level of IL-12,IFN-γ,TNF-α and IL-10 increased significantly from week 4 to week 6,week 2 to week 8,week 2 to week 6 and week 4 to week 12,respectively,reaching the highest level(25.0±5.8)ng/L on week 4,(575.0±28.9)ng/L on week 2,(50.0±11.5)ng/L on week 2 and (42.5±2.9)ng/L on week 8,respectively,as compared with the values on week 0[(11.3±2.5),(125.0±28.9),(11.3±2.5),(12.5±2.9)ng/L,all P < 0.01]; in the intranasal immunization group,it was similar about the values of IL-12,IFN-γ,TNF-α and IL-10 could be seen from week 4 to week 6,week 2 to week 10,week 4 to week 10 and week 6 to week 16,respectively,reaching the highest level(25.0±5.8)ng/L on week 6,(725.0±28.9)ng/L on week 4,(27.5±2.9)ng/L on week 6 and (60.0±11.5)ng/L on week 6,respectively,as compared with the values on week 0[(11.3±2.5),(125.0±28.9),(11.3±2.5),(12.5±2.9)ng/L,all P < 0.01]. The cytokine levels in the groups with EgAg,ConA or LPS stimulus were significantly higher than those in the corresponding splenocyte suspension groups(P < 0.05 or < 0.01),and the cytokine levels in the groups with ConA or LPS stimulus were obviously higher than those in the corresponding groups with EgAg stimulation (P < 0.05 or < 0.01). Conclusion The mixed responses of Th1 and Th2 types can be induced in mice immunized with the transgenic alfalfa in the early period post immunization(2-10 weeks).

Adult ; Aged ; Aged, 80 and over ; Antifungal Agents ; therapeutic use ; Female ; Hematologic Neoplasms ; drug therapy ; microbiology ; Humans ; Male ; Middle Aged ; Mycoses ; complications ; drug therapy ; Pyrimidines ; therapeutic use ; Treatment Outcome ; Triazoles ; therapeutic use ; Voriconazole ; Young Adult

Objective To construct a prokaryotic expression vector of cystatin C (Cys C), purify Cys C protein produced by the expression system, and prepare its antiserum. Methods Total RNA was isolated from HL-60 cells, and human Cys C gene was amplified with RT-PCR. The cDNA fragment was cloned into pMD18-T vector and which was confirmed by sequencing. The enzyme-digested target fragment was cloned into PET-32(a) expression vector and transfected into E.coli. BL 21(DE3), in which Cys C expression was induced. After the inclusion body protein was purified through Ni2+ affinity chromatography, processed by dialysis, identified by Western blotting, a rabbit was immunized with the fusion protein, and the antiserum was obtained. Results The result of DNA sequence analysis showed that the cloned Cys C gene sequence was completely corresponding to GenBank data. SDS-PAGE and Western blotting showed that the expressed Cys C fusion protein was about 35?103, mainly existing in the inclusion body of E.coli., that could be purified through Ni2+ affinity chromatography. The titer of the antiserum to the purified protein was 1∶8 000 by ELISA, and Western blotting confirmed that the antiserum reacted specifically to the Cys C protein. Conclusion A recombinant Cys C protein and the specific polyclonal antibody have been obtained, which provides a basis for establishment of immunoassays of human Cys C.

Asthma ; diagnosis ; Child ; Humans ; Perception

Fluorescence enhancement reaction of flavoxate hydrochloride (FX) in strong alkali solution was studied, the mechanism of the reaction was investigated, and a novel fluorimetric method for analysis of FX in drug sample was established. FX has no intrinsic fluorescence, but it can slowly produce fluorescence in strong alkali solution. Heating can promote the fluorescence enhancement reaction. In 3D fluorescence spectra of the decomposition product of FX, two fluorescence peaks, located respectively at excitation wavelengths λex/ emission wavelength λem =223/410 nm, and 302/410 nm, were observed. Using quinine sulfate as a reference, fluorescence quantum yield of the decomposition product was measured to be 0.50. The structural characteriza- tion and spectral analysis of the decomposition product reveal that ester bond hydrolysis reaction of FX is firstly occurred during heating process, forming 3-methylflavone-8-carboxylic acid (MFA), then a cleavage reaction of the γ-pyrone ring of MFA occurred, producing α, β-unsaturated ketone. This product includes adjacent hydroxyl benzoic acid group in its molecule, which can form intramolecular hydrogen bond under alkaline condition, so that increase the conjugate degree and enhance the rigidity of the molecule, and thereby cause fluorescence enhancement. Based on this fluorescence enhancement reaction, a fluorimetric method was proposed for the determination of FX. A linear calibration curve covered the concentration range 0.020 3-0.487 µg · mL. The regression equation was I(F) = 23.9 + 5357.3 c, with correlation coefficient r = 0.999 7 (n = 8), detection limit D = 1.1 ng · mL(-1). The method was applied to the analysis of FX tablets, with a spiked recovery rate of 100.2%. The reliability of the method was verified by a UV-spectrophotometric method.
Alkalies ; Calibration ; Chemistry, Pharmaceutical ; Flavoxate ; analogs & derivatives ; chemistry ; Fluorescence ; Limit of Detection ; Reproducibility of Results ; Solutions ; Tablets