A rapid, reliable and sensitive pressurized capillary electrochromatography-Laser induced fluorescence ( pCEC/LIF ) method with trifluoroacetic acid ( TFA ) pre-column derivation for simultaneous determination of four aflatoxin ( AFB1 , AFB2 , AFG1 , AFG2 ) was developed. This method included separation on a capillary column packed with 1. 8μm C18 particles using 0. 05% FA aqueous solution/methanol (55:45, V/V) as mobile phase at a pump flow rate of 0. 05 mL/min when the split ratio was 1:300. Under the optimum conditions including running voltage of 15 kV, excitation wavelength of 375 nm and emission wavelength of 450 nm, the baseline separation of four aflatoxins was achieved within 10 minutes. The limits of detection (LODs) were 0. 02, 0. 016, 0. 008 and 0. 01 μg/L for AFG1, AFB1, AFG2, AFB2(S/N=3), respectively. The linear detection ranges of AFG1 , AFB1 , AFG2 , AFB2 were 0. 1-10, 0. 1-10, 0. 1-3 and 0. 1-3 μg/L with correlation coefficients (R2) of 0. 9999, 1. 0000, 0. 9995 and 0. 9997, respectively. The established method was applied to analyze the peanut butter, and the recoveries of standard addition experiment were between 90 . 0% and 112 . 0% for all analytes ( RSDs=0 . 5%-1 . 9%) .

Animals ; Bone Marrow Cells ; cytology ; Cells, Cultured ; Culture Media, Conditioned ; Megakaryocytes ; cytology ; Mice ; Mice, Inbred Strains ; Thrombopoietin ; pharmacology

Objective An interaction network among PD-1-ligands and other proteins were built to add deeper understanding of PD-1 signal pathway and to supply the theoretical data on the clinical application study of PD-1 and its ligands.Methods Searching the literature about PD-1 and its ligands in the PubMed,the result of which would be used to summarize the proteins that had been reported to have interactions with PD-1 and its ligands,and then,quadratic search would be performed on these proteins.Finally,the software Cytoscape would be used to build and analyze the interaction network and verified by PCR.Results There were 122 and 126 nodes in the PD-1/PD-L1 network and PD-1/PD-L2 network respectively.These proteins were involved in TCR signal path-way,cell adhesion,JAK-STAT signal pathway and interaction between cytokines.Meanwhile,a perspective that PD-L1 may influ-ence on CXCR4 through JAK-STAT pathway based on the network which had been supported by qPCR.Conclusion The bioinfor-matics suggested that there are differences between the mechanism of PD-1/PD-L1 and PD-1/PD-L2.Additionally,the method, which was based on the literature mining and bioinformatics,is good to put the biological literature to rational utilization and to pro-vide guidance for experiments.

Insulin resistance model of 3T3-L1 adipocytes were prepared with plamotic acid.Adipocytes with generated insulin resistance were cultured with different concentrations of globular domain of adiponectin(gAd:250,500,1 000 ng/ml).The cell culture medium glucose content was detected with the glucose oxidase method,the mRNA expressions of insulin receptor substrate-1 (IRS-1),phosphatidylinositol-3 kinase(PI3K),and protein kinase B(PKB) were detected with real-time quantitative PCR method.The phosphorylation of IRS-1 was detected by Western blot.Compared with the control group,the experimental group showed significantly increased glucose consumption (P ＜ 0.01),and with the increasing gAd concentration,glucose consumption was gradually increasing.IRS-1 phosphorylation was increased gradually with the increasing concentration of gAd.These results suggest that gAd can promote glucose uptake by 3T3-L1 adipocyte model with generated insulin resistance.This may be correlated with promoting insulin signal transduction and improving insulin resistance in adipocytes.

Cytoplast and microprotoplast are main subprotoplasts that can play an important role in plant genetic improvement.The present review highlights the advancements in isolation and fusion of plant subprotoplast,and some suggestions and prospects are proposed for the future studies.

Objective To evaluate the disinfection effect of alginate impressions using three- step flushing approach with alkaline electrolyzed- oxidizing water (AEOW), neutral water, acidic electrolyzed-reductive water (AERW) in sequence, and therefore explore a convenient, safe and efficient approach for disinfection of impressions. Methods 60 pairs of alginate impressions of left maxillary dentition and right maxillary dentition were taken from 60 patients set the left ones and the right ones into the experimental group and the control group randomly. For the experimental group, the impressions were flushed with AEOW, neutral water and AERW in sequence, for the control group, the impressions were flushed with water. Then the disinfection effect of the two groups at different flushing time was compared. Results The number of bacterial on the alginate impressions was reduced in both groups at 3, 6 and 9 minutes. The number of bacterial of the experimental group was lower than that of the control group at the same time point, the differences were statistically significant,Z values were -6.160, -6.159, -5.920 respectively, P<0.01. The disinfection effect of the three- step flushing approach was 80.29%, 92.91%, 97.98% at 3, 6 and 9 minutes respectively. The killing rate of the experimental group was higher than that of the control group at the same time,the differences were statistically significant, χ2 values were 13.51, 12.11, 8.31 respectively, P<0.01. Conclusions The disinfection effect of the three - step flushing approach for 6 minutes can reach a satisfactory level. Hence, the three- step flushing approach is a convenient, safe, and efficient approach for disinfection of impressions.

Objective: To identify neuroprotective extracts with the protective effects on Aβ25-35-induced SH-SY5Y cell injury via high content screening (HCS). Methods: Hoechst 33342/PI double staining method was used to screen neuroprotective extracts from 60 Chinese materia medica (CMM) extracts. Further more, the effects of neuroprotective extracts on Aβ25-35-induced changes in the levels of Caspase-3/7 were detected. Results: The results showed that 17 extracts had obviously neuroprotective effects. Among the 17 extracts, 8 of them inhibited Aβ25-35-induced up-regulation of Caspase-3/7. Conclusion: HCS is an efficient method to screen neuroprotective extracts with the protective effects of Aβ25-35-induced SH-SY5Y cell injury. The neuroprotective extracts have potential medicinal value in Alzheimer's disease.

OBJECTIVETo explore the clinical efficacy and action mechanism of acupuncture for premature ovarian failure (POF).

METHODSAccording to prospective cohort study design, 30 cases were included. Based on theory of regulating Chong and Conception Vessels as well as soothing the liver and calming the nerves, acupuncture at Guanyuan (CV 4), Dahe (KI 12), Taixi (KI 3), Taichong (LR 3), Baihui (GV 20) was applied, three times per week and 3 months were considered as one session. Totally two sessions were performed. The menstruation condition, estradiol (E2), follicle-stimulating hormone (FSH) and scores of the clinical perimenopausal symptoms were taken as treatment outcomes.

RESULTSThe total effective rate was 86.7% (26/30) and the cured rate was 16.7% (5/30). The recovery rate of menstruation was 16.7% (5/30) and the regain rate of menstruation was 56.7% (17/30). After the treatment, the level of E2 was significantly increased from (45.41 +/- 18. 09) pmol/L to (59.07 +/- 24.21) pmol/L (P < 0.05), and the score of perimenopausal symptoms was obviously reduced from 14.28 +/- 8. 30 to 4.04 +/- 3.28 (P < 0.01). However, no statistical differences could be seen in FSH before and after treatment (P > 0.05).

CONCLUSIONAcupuncture has certain effect on improving menstruation and perimenopausal symptoms in POF patients, which is likely to he related with increasing the level of E2.

Acupuncture Therapy ; Adolescent ; Estradiol ; metabolism ; Female ; Humans ; Luteinizing Hormone ; metabolism ; Primary Ovarian Insufficiency ; metabolism ; therapy ; Prospective Studies ; Young Adult

AIM:To observe related biological parameters of 3 minutes dark-room provocative test in patients with laser peripheral iridectomy(LPI) in the fellow eyes of acute primary angle-closure (APAC) by ultrasound biomicroscopy (UBM).To explore the risk factors in primary angle closure suspect(PACS) patients with progressive angle closure after LPI.METHODS: Seventy-eight eyes of APAC patients without peripheral anterior synechia were selected.Each eye underwent 3 minutes dark-room provocative test after LPI.Anterior segment parameters, including anterior chamber depth (ACD), anterior chamber angle open distance500 (AOD500), peripheral iris thickness (PIT), iris convex (IC), the position of iris insertion and trabecular-ciliary process distance (TCPD), and the number of positional angle closure(NPAC) were observed and analyzed by statistic methods.RESULTS:Patients with APAC were examined by UBM after LPI and 26 eyes(33%) occurs at least one positional angle closure,19 eyes(24%)were positive in 3 minutes dark-room provocative test among them.It occurs a positive relationship between the elevation intraocular pressure and the number of positional angle closure in dark-room provocative test(r=0.84, P<0.01).AOD500, IT and IC were significantly changed from normal light to darkroom between positional angle closure positive group and positional angle closure negative group(all P<0.01).In single factor analysis, AOD500(P=0.003), IT(P=0.012), IC(P=0.043), TPCD(P=0.015), the position of iris insertion(P=0.024) were correlative factors of positive results.In multiple-factor analysis, only IT(P=0.011), TPCD(P=0.009), iris root attachment points(P=0.02) were independent risk factors of positive results.CONCLUSION:A certain proportion of patients with PACS after LPI appeared positional angle closure in a dark room.Peripheral iris hypertrophy, anterior displacement of the ciliary body and iris root attachment points are vital risk factors.Long-term follow-up study and intervention treatment are required in these patients after LPI.

Objective To study the effects of oxymatrine as inhibitor of hemorrhagic fever with renal syndrome virus (HFRSV) infection in vitro and in vivo.Methods In vitro studies,a dose of oxymatrine without cytotoxicity and 76-118 strain of HFRSV was taken to treat Vero cells in three ways:①After treated with oxymatrine for 48 h,Vero cells were attacked by HFRSV at dilution of 10-1 ～ 10-6,respectively for 24 h before changing to maintenance medium; ②Vero cells were first attacked by HFRSV of 10-1 ～ 10-6 dilution respectively,then oxymatrine was used for 48 h before changing to maintenance medium; ③Vero cells were attacked by HFRSV at dilution of 10-1 ～ 10-6 respectively,and meanwhile treated with oxymatrine for 48 h before changing to maintenance mcdium.Each dilution handled four porocytes,and four positive controls were set up at the same time.Indirect immunofluorescence assay (IFA) was performed to determine the inhibitory effect of oxymatrine in experimental group and positive control.In vivo studies,thirty 2-week-old hamsters,weighing about 30-40 g,were divided into experimental and control groups according to body weight,n =15.These aninals were inoculated intraperitoneally with HFRSV in 100TCID50(0.1 ml each); on days 4-13,0.1 ml of oxymatrine 1:100 were given to each hamster in experimental group daily by intraperitoneal injection,while the same amount of saline was given to the control ones.Lung tissue of hamsters was then dissected out to slice to be identified by immunofluorcscence stain.Results It was demonstrated that oxymatrine with the diluted fractions of 1:8 was safe in vitro.When the virus dilution of HFRSV was l0-4,compared with control groups,the differences were statistically significant in method 2 and 3 (z =-2.53,-2.53,all P ＜ 0.05),while no statistical significance in method 1 (z＝5.36,P＞ 0.05).When the virus dilution of HFRSV was 10-1 ～ 10-3,10-5,10-6,the differences were not statistically significant (z--0.00,-0.32,-0.19,4.21,4.21,all P ＞ 0.05).In vivo studies,compared with control group,the differences were statistically significant in experimental group (z =-3.85,P ＜ 0.05).Conclusion Oxymatrine significantly inhibites HFRSV.