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).

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.

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.

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).

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 investigate the mechanism of effects of acitretin on the treatment of psoriasis. Methods The expression of vascular endothelial growth factor (VEGF) and micro-vascular density (MVD) were detected by immunohistochemical technique before and after treatment with acitretin in 32 patients with psoriasis. Serum level of VEGF was measured by double antibody sandwich enzyme-linked immunosorbent assay (ELISA). Results The expression of VEGF protein and MVD were significantly higher in psoriatic lesions before treatment with acitretin than those after treatment (P

Objective To establish a sensitive and specific liquid chromatography method for the determination of loganin and morroniside in rat serum containing Liuwei Dihuang Formula(LDF).Methods Loganin and morroniside were extracted from the rat serum using SPE column,then separated on a C_(18)column.A HPLC method for determining serum concentration of mor- roniside and loganin in rats before and after administration LDF was established.Results A good linear calibration curve of lo- ganin and morroniside was obtained.The recovery of the method was in the range of 95 %～105 %.The extracted recovery was more than 75 % ,and within-day and intra-day precisions were less than 12 %.Conclusion This method is sensitive, simple and rapid for monitoring of drug level in the study on pharmacokinetics and different combination of LDF.

Asthma ; diagnosis ; Child ; Humans ; Perception