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Objective To discuss the effect of H2 on osteoclast differentiation of RAW264.7 cells induced by RANKL and RANKL/TNF-α.MethodsRAW264.7 cells were treated with H2 in the presence of RANKL and RANKL/TNF-α.RAW264.7 cells viability was assessed by CCK-8.Test the Oxidative Stress of the induced RAW264.7.The number of TRAP-positive cells were counted under light microscopy.The levels of cathepsin K (CTK) and matrix metalloprotein-9(MMP-9) mRNA were analyzed by real-time PCR.ResultsH2 can not influence the RAW264.7 cell viability but can lower oxidative stress.The significant difference(P<0.05) indicated that H2 could significantly decrease the number of TRAP-positive MNCs.The significant difference among the 4 groups in CTK and MMP-9 genes (P<0.05) indicated that H2 could down-regulate their mRNA expression.ConclusionH2 can reduce the oxidative stress and inhibit differentiation of RAW264.7 cells into osteoclasts.
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Objective To explore a new outpatient mode for pregnancy nutrition to adapt to digital hospital.Methods The outpatient management mode and methods were analyzed for pregnancy nutrition.A new outpatient mode combining the technologies of mobile internet and remote monitoring was developed with consideration on standardization,and the effect of the new mode was discussed on pregnancy nutrition outpatient.Results Mobile internet technology and remote monitoring technology contributed to enhancing the efficiency of pregnancy nutrition outpatient,and facilitated the nutrition service of common pregnant women as well as the precision and individualized nutrition management of high-risk ones such as those with gestational diabetes mellitus.Conclusion The new outpatient mode enhances the doctor's efficiency and pregnancy care,and thus is worthy promoting practically.
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AIM To study the chemical constituents from Tibetan medicine Tinospora sinensis (Lour.)Merr..METHODS The ethyl acetate fraction of ethanol extract from T.sinensis was isolated and purified by silica,Sephadex LH-20,preparative HPLC column and recrystallization,then the structures of obtained compounds were identified by physicochemical properties and spectral data.RESULTS Nine compounds were isolated and identified as rutin (1),quercetin (2),kaempferol (3),luteolin (4),myricetin (5),paeonol (6),N-cisferuloyltyramine (7),myricetin-3-O-β-D-glucopyranoside (8),quercetin-3-O-α-L-rhamnoside (9).CONCLUSION All the compounds are isolated from this plant for the first time.
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AIM To study the chemical constituents from Tibetan medicine Tinospora sinensis (Lour.)Merr..METHODS The ethyl acetate fraction of ethanol extract from T.sinensis was isolated and purified by silica,Sephadex LH-20,preparative HPLC column and recrystallization,then the structures of obtained compounds were identified by physicochemical properties and spectral data.RESULTS Nine compounds were isolated and identified as rutin (1),quercetin (2),kaempferol (3),luteolin (4),myricetin (5),paeonol (6),N-cisferuloyltyramine (7),myricetin-3-O-β-D-glucopyranoside (8),quercetin-3-O-α-L-rhamnoside (9).CONCLUSION All the compounds are isolated from this plant for the first time.
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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.
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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).
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Objective To cultivate and identify the transgenic affalfa containing Echinococcus granulosus Eg95 gene. Methods The alfalfa plants were transformed by co-cultivating alfalfa cotyledons via recombinant Agrobacterium tumefaciens LBA4404 harboring pBI-Eg95. The transgenic alfalfa explants were selected by kanamyein after calli formation, shoots and roots regeneration in the selective medium, the seedlings of transgenic plants were obtained which were finally transplanted into pots containing nutrient soil. After 2-3 months growth, the complete transgenic alfalfa plants containing Echinococcus granulosus Eg95 gene were obtained. To identify the transgenic alfalfa plants, the total DNA, RNA and leaf protein were extracted from fresh leaf tissue of the transgenic alfalfa plants and confirmed by PCR, RT-PCR, SDS-PAGE and Western blot assay. Results A specific band around 471 bp was amplified by PCR with total DNA, and the same band was obtained by RT-PCR with total RNA, which confirmed that the Eg95 gene was stably integrated into the transformed alfalfa genome. SDS-PAGE analysis showed that the relative molecular mass(Mr) of the expressed protein was about 16.5×103, consistent with the Eg95 protein, and the level of Eg95 expression was up to 0.06% of total soluble leaf protein by Bio-Rad Quantity one assay. Western blot verified the expressed protein was reactive with the sera of mice infected with Echinococcus granulosus. Conclusion The transgenic alfalfa plants containing Echinococcus granulosus Eg95 gene are successfully cultivated.
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To investigate the difference in absorptive of tetrahydropalmatine (THP) and l-tetrahydropalmatine (l-THP) in rat intestine as well as the mechanism of the absorption of THP, in situ single pass perfusion model was used and the concentration of THP in perfusate was determined by HPLC. The absorption rate constant (k(a)) and effective permeability values (P(eff)) of THP had no significant difference (P > 0.05) at concentration of 8, 16 and 32 microg x mL(-1) in perfusion or in four different regions of intestine of rat (duodenum, jejunum, ileum, colon). The absorption of l-THP and THP in jejunum had significant difference (P < 0.05). The k(a) and P(eff) of THP increased obviously when verapamil was co-perfused with THP, while those of l-THP were not influenced by verapamil. The absorption of THP in intestine showed the passive diffusion process, and without a special absorption region. The stereoselective absorption difference may result from stereoselective combination of P-glycoprotein with d-THP.
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Animals , Female , Male , Rats , ATP Binding Cassette Transporter, Subfamily B, Member 1 , Berberine Alkaloids , Chemistry , Pharmacokinetics , Colon , Metabolism , Duodenum , Metabolism , Ileum , Metabolism , Intestinal Absorption , Jejunum , Metabolism , Perfusion , Permeability , Rats, Sprague-Dawley , Sex Factors , Stereoisomerism , Verapamil , PharmacologyABSTRACT
The purpose of this study is to assess the maternal iron status during pregnancy and to evaluate the relationships between the iron indices of maternal, umbilical cord serum, placenta and pregnancy outcomes. Venous bloods samples were drawn from 54 pregnant women just before delivery and cord bloods of their newborn babies were collected immediately after birth. And also, placental tissues were extracted. We investigated the difference of the iron status indices of maternal, umbilical cord serum and placental tissue between two gestational age group (PT group, NT group : preterm delivery and normal term delivery at 34.9wk and 39.0wk of mean gestational length, respectively) and also assessed correlations of iron status indices of maternal, umbilical cord serum and placenta tissue. And lastly, we related between birth weight and iron status indices of maternal, umbilical cord serum and placental tissue. The concentrations of maternal serum ferritin and of placental iron were significantly higher in PT group (32.1 +/- 21.1 ng/ml, 68.5 +/- 16.7microgram / g), than those of NT group (20.8 +/- 11.6 ng / ml, 53.2 +/- 17.4 microgram / g) respectively (p < 0.001). However the serum ferritin of umbilical cord were significantly higher in NT group (PT : 109.4 +/- 65.7 ng/ml, NT : 147.0 +/- 56.8 ng / ml) than those of PT group (p < 0.05). Our results showed that a negative association between birth weight (r=-0.361) and maternal serum ferritin and that a positive association between birth weight and umbilical cord serum ferritin (r=0.261). Despite not a significant difference, there was tendency that highest concentration of maternal serum ferritin was associated with the lowest birth weight. These findings indicate that birth weight of newborn is dependent of multiple factors such as maternal iron status during pre-pregnancy, body size, general nutritional status. Although for women who enter pregnancy with low iron stores, enough intakes of iron during pregnancy could produce undesirable pregnancy outcome. Therefore we suggest for successful pregnancy outcome and delivery differential iron supplementation program will be carried out individual pregnant women on the basis of pre-pregnancy nutritional status.
Subject(s)
Female , Humans , Infant, Newborn , Pregnancy , Birth Weight , Body Size , Ferritins , Fetal Blood , Gestational Age , Iron , Nutritional Status , Parturition , Placenta , Pregnancy Outcome , Pregnant Women , Umbilical CordABSTRACT
Retinal detachment is a rare disease in children but cases of serous retinal detachment due to use of systemic corticosteroids have been reported in and out country. Rhematogenous retinal detachment has developed in a child with oral and steroid pulse therapy due to nephrotic syndrome. He was treated with laser and buckling in right and left eye respectively. Currently he is under observation through the out patient department.