AIM： To estabish a HPLC assay for the determination of pirarubicin（Pir） in plasma.METHODS： Daunomycin（DM） was used as the internal standard.Plasma samples were extracted with CHCl3∶ CH3OH（2∶ 1） .0.4M NH4Cl buffer（pH=9.0） solution： CH2OH（1∶ 9） and the internal standard were added.Separation was carried out on a 250mm× 4.6mm（5μ m） phenomenex column with 0.04M KH2PO4（pH=3.0） ∶ CH3CN（68∶ 32） as mobile phase.Fluorescent detector was set at an excitation wavelength of 480nm and an emission wavelength of 550nm.RESULTS： The calibration curves for serum Pir was linear over the range of 5～1 000ng/ml（r=0.9 997） .The recovery of Pir was 95.3％ .The within-day and between-day variations were less than 5％ .T1/2β ， CLs， Vd and AUC of Pir were（12.8± 5.9） h， （128.3± 52.6） L/（m2· h） ， （1 754.3± 478.2） L/m2 and （428.7± 137.2） ng/（h· ml） ， respectively.CONCLUSION： The method is suited for monitoring blood concentration and pharmacokinetic study of Pir.

Objective To construct eukaryotic expression vector of human antimicrobial peptide LL-37 pPIC9-LL-37, and transform the plasmid pPIC9-LL-37 into P.pastoris GS115 to obtain the recombinant P.pastoris strains.Methods The full-length of antimicsobial peptide LL-37 gene was artificially synthesized by overlap extension method and was fused to pPIC9 and then the fused plasmid was transformed into E.coli DH5?.After analysis by PCR and sequencing,the plasmid pPIC9-LL-37 was transformed into P.pastoris.The colonies exhibiting the phenotype of His+Mut+ or His+Mut-were screened by means of MM and MD plates and the insertion was confirmed by PCR.Results The results of PCR and sequencing confirmed that the LL-37 gene was correctly inserted into pPIC9. The colonies of 10 His+Mut+ and 9 His+Mut-were obtained by means of MM and MD plates screening and were confirmed by PCR.Conclsion The recombinant P.pastoris strains containing LL-37 were successfully obtained.

Objective: Dithiothreitol(DTT) and ?-Mercaptoethanol(2-ME) are important reducing agents for SDS-PAGE.This study is to observe the diffusing effect of DTT and 2-ME during electrophoresis,and to find a way of avoiding this effect.Methods: We placed protein samples containing reducing agents and non-reduced protein samples separately in the sample wells at intervals for SDS-PAGE electrophoresis,and determined whether or not the electrophoretic lanes of the non-reduced samples were interfered by the adjacent lanes.Results: DTT and 2-ME diffused to the neighboring lane,so that the non-reduced samples were reduced partially.The spreading effect was positively correlated with the content of the reducing agent.Conclusion: DTT and 2-ME have a diffusing effect during SDS-PAGE electrophoresis.In separating the reduced and non-reduced proteins in the same gel at the same time,at least a blank lane should be set up in between them in order to avoid the diffusing effect.

Objective:To evaluate the effects of dexamethasone on systemic lupus erythematosus complicated with cognitive dysfunction.Methods:Ten wild type mice and 20 MRL/lpr mice were applied for the research.MRL/lpr mice were randomly assigned to a MRL/lpr group and a MRL/lpr + dexamethasone (1.5 mg/kg) group.Interleukin-6 (IL-6),IL-1β,and tumor necrosis factor alpha (TNF-α) in serum and hippocampus were detected.The protein phosphorylation levels of phosphoinositide 3-kinase (P-PI3K),protein kinase B (P-Akt),NF-kappa-B inhibitor alpha (P-IκBa) and nuclear transcription factor kappa-B p65 (P-NF-κB p65) were detected by Western blot,the level of P-NF-κB p65 also was detected by immunohistochemistry.Results:Treatment with dexamethasone (1.5 mg/kg) alleviated the cognitive dysfunction and decreased the levels of IL-6,IL-1 β and TNF-α in serum and hippocampus,and reduced the levels of P-PI3K,P-Akt,P-IκBa and P-NF-κB p65 in hippocampus in MRL/lpr mice.Conclusion:Dexamethasone may play a protective role in the cognitive function by decreasing the levels of TNF-α and IL-1 β in the hippocampus of MRL/lpr lupus mice.

Objective To understand the variation in response of Oncomelania hupensis to niclosamide. Methods Snails were collected from 37 sampling areas distributed in 10 provinces (municipalities) using random environmental sampling methods in accordance with the different types and categories of snail habitats. In laboratory the snails were immersed in solutions of niclosamide for 24 and 48 hours at 25℃. Results 1.0 mg/L niclosamide showed 100% killing effect on snails in 24 hours. The LC 50 concentrations for snails immersed for 24 hours ranged from 0.0320 to 0.1689 mg/L with a mean value of 0.0920 mg/L. 0.5 mg/L niclosamide showed 100% killing effect on snails in 48 hours. The LC 50 values for snails immersed for 48 hours ranged between 0.0299 and 0.1114 mg/L with a mean of 0.0627 mg/L. There is a significant difference in snail sensitivity to niclosamide between sampling areas. Conclusion The sensitivity to niclosamide varied in snails from different sampling fields, but the chemical in a concentration of 1.0 mg/L showed 100% effect of killing snails, which is consistent to the manual of schistosomiasis control.

Digital healthcare empowers optimization of medical services.This article introduced the practices and achievements of Hangzhou in the following aspects: the construction of a hierarchical medical system guided by " people-oriented integrated service" ; development of the five digital platforms, namely regional health information platform, integrated medical care platform, remote consultation platform, intelligent supervision platform and doctor-patient remote interaction platform; development of " Smart Medicine" to promote institutional cooperation, service integration, management coordination, quality improvement and doctor-patient interaction.The paper also analyzed the supporting role of digital healthcare construction in promoting the hierarchical medical system, and put forward corresponding countermeasures and suggestions.

OBJECTIVE: To study the role of Nrf2/ARE signaling pathway in cypermethrin-induced oxidative stress and apoptosis of cerebral cortex neurons in C57BL/6 mice. METHODS: The cortical neurons of C57BL/6 mice were cultured and identified, and a cypermethrin-induced cell injury model was established by treating the cells with 0, 25, 50 and 100 μmol/L of cypermethrin for 48 h. CCK-8 assay was used to analyze the effects of cypermethrin on the cell viability, and the fluorescence probe DCFH-DA was used for detecting intracellular reactive oxygen species (ROS); flow cytometry was performed for determining the apoptosis rate of the cells. The mRNA and protein expression levels of Nrf2 and its downstream genes HO-1 and NQO1 were detected using qPCR and Western blotting. RESULTS: Exposure to cypermethrin at different doses inhibited the viability of the cultured cortical neurons. With the increase of cypermethrin dose, the viability of the neurons decreased progressively, the intracellular ROS and the cell apoptosis rate increased, and the neuronal injury worsened. At the dose of 50 and 100 μmol/L, cypermethrin significantly down-regulated the expressions of HO-1, NQO1 and Nrf2 at both the mRNA and protein levels in the cells ( < 0.01). CONCLUSIONS Cypermethrin exposure shows a dose-dependent neurotoxicity by inhibiting Nrf2/ARE signaling pathway, down-regulating the expression of Nrf2 and its downstream genes HO-1, NQO1 mRNA and protein, and inducing oxidative damage and apoptosis in primary mouse cortical neurons, .
Animals ; Carboxylic Ester Hydrolases ; Cerebral Cortex ; Mice ; Mice, Inbred C57BL ; NF-E2-Related Factor 2 ; Neurons ; Pyrethrins ; Signal Transduction