Air ; analysis ; Hydrobromic Acid ; analysis ; Ion-Selective Electrodes ; Workplace

OBJECTIVETo establish the quality standard of Entadae Semen, and provide scientific basis for its quality control.

METHODTLC and HPLC were used for qualitative identification and quantitative analysis of phaseoloidin and entadamide A-O-β-D-glucopyranoside in Entadae Semen. The test of water content, ash and ethanol-soluble extractives of Entadae Semen was carried out according to the methods recorded in appendix of Chinese Pharmacopeia (2010 edition).

RESULTThe TLC was well separated with clear spots. The linear range of phaseoloidin was between 0.014 to 2.747 g x L(-1) (r = 0.999 6, n = 9) with an average recovery rate of 101.06% (RSD 0.90%, n = 6); the linear range of entadamide A-O-β-D-glucopyranoside was between 0.002 to 0.452 g x L(-1) (r = 0.999 7, n = 9) withan average recovery rate of 101.52% (RSD 1.09%, n = 6). The content of phaseoloidin in sample is between 5.12% to 9.24%, entadamide A-O-β-D-glucopyranoside is between 0.55% to 2.17%, alcohol-soluble extracts is between 30.9% to 45.2%, water is between 6.6% to 8.6%, and total ash is between 2.4% to 2.9%.

CONCLUSIONThe established standard is acceptable for quality control of Entadae Semen.

China ; Chromatography, High Pressure Liquid ; Chromatography, Thin Layer ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; standards ; Fabaceae ; chemistry ; Quality Control

Adolescent ; Asthma ; blood ; Child ; Child, Preschool ; Cough ; blood ; Eosinophil Cationic Protein ; blood ; Female ; Humans ; Immunoglobulin E ; blood ; Infant ; Male

Argyrogramma agnata has been selected as a substitutive host insect for producing Dendrolimus punctatus Cytoplasmic Polyhedrosis Virus (DpCPV). In our experiment, it is very susceptible to DpCPV. The DpCPV produced in A. agnata is designated Aa-DpCPV. The cytoplasmic polyhedra body (CPB), the virion size and the shape of Aa-DpCPV are same as that of its original DpCPV (DpCPV-W1984). The RNA bands of Aa-DpCPV and DpCPV-W1984 all have 10 RNA segments respectively in 3% PAGE, which molecular weights ranged in size from 2.98×106 to 0.66×106 Dalton. Aa-DpCPV has the same strong toxicity as that of DpCPV-W1984 (from D. punctatus) to D. punctatus (Walker) larva. So it can be applied to the pine caterpillar control. The DpCPV yield in A.agnata is 2.5×108CPB/larva.

Penaeidin-2 (Pen-2) is an important antimicrobial peptide derived from the Pacific white shrimp, Penaeus vannamei, and possesses both antibacterial and antifungal activities. Recent studies suggest that recombinant penaeidins show similar activities to the native Pen-2 protein. Previous researches have shown that some antimicrobial peptides (AMPs) exhibit cytotoxic activity against cancer cells. To date, there have been no studies on the antitumor effects of Pen-2. This study evaluated the potential of recombinant pen-2 (rPen-2) in the selective killing of kidney cancer cell lines ACHN and A498, and its action mechanism. MTT assays found the maximal growth inhibition of HK-2, ACHN and A498 cells treated with 100 μg/mL rPen-2 at 48 h was 13.2%, 62.4%, and 70.4%, respectively. DNA-specific fluorescent dye staining showed a high percentage of apoptosis on cancer cells. Flow cytometry revealed that the apoptosis rate of HK-2, ACHN and A498 cells was 15.2%, 55.2%, and 61.5% at 48 h respectively, suggesting that rPen-2 induced higher apoptosis rate in cancer cells than in HK-2 cells. Laser confocal scanning microscopy demonstrated that the plasma membrane was the key site where rPen-2 interacted with and destroyed tumor cells. Scanning electron microscopy showed the morphologic changes of the cell membranes of kidney cancer cells treated with rPen-2. These results suggest that rPen-2 is a novel potential therapeutic agent that may be useful in treating kidney cancers.

This paper discusses the relationship between stimulating pulse width and the threshold of electrically evoked compound action potential (ECAP). Firstly, the rheobase and chronaxy from strength-duration curve of nerve fiber was computed using the shepherd's experiment results. Secondly, based on the relationship between ECAP and the action potential of nerve fiber, a mathematical expression to describe the relationship between stimulating pulse width and ECAP threshold was proposed. Thirdly, the parameters were obtained and the feasibility was proved to the expression with the results of experiment using guinea pigs. Research result showed that with ECAP compared to the action potential of nerve fiber, their threshold function relationship with stimulating pulse width was similar, and rheobase from the former was an order smaller in the magnitude than the latter, but the chronaxy was close to each other. These findings may provide meaningful guidance to clinical ECAP measurement and studying speech processing strategies of cochlear implant.
Action Potentials ; Animals ; Auditory Threshold ; Cochlear Implantation ; Cochlear Implants ; Electricity ; Evoked Potentials ; Guinea Pigs ; Neural Conduction

OBJECTIVETo explore the effect of ERK1/2 MAPK pathway on the expression of Kv1.5 channel, a voltage-gated potassium ion channel, in rat pulmonary artery smooth muscle cells (PASMCs) and its mechanisms during the process of hypoxia.

METHODSThe PASMCs derived from SD rats were cultivated primarily. The third to sixth generation of PASMCs were divided into 5 groups randomly: (1) Normal group (N); (2) Hypoxic group (H); (3) Demethy sulfoxide(DMSO) group (HD); (4) U0126 group (HU): 10 micromol/L U0126; (5) Anisomycin group (HA): 10 micromol/L anisomycin. There were three dishes of cells in each group. The cells in normal group were cultured in normoxic incubator (5% CO2, 37 degrees C), the cells in other groups were added to 0.05% DMSO in the hypoxic incubator (5% CO2, 2% O2, 37 degrees C), all cells were cultured for 60 h. RT-PCR and Western blot were used to detected the espressions of Kv1.5 mRNA and protein in PASMCs.

RESULTSCompared with N group, the expressions of Kv1.5 mRNA and protein in H, HD and HA groups were reduced significantly (P < 0.05); Compared with H group and HD groups, Kv1.5 mRNA and protein expressions in HU group were increased sharply (P < 0.05). Compared with the HU group, Kv1.5 mRNA and protein expressions in HA groups were significantly lower (P < 0.05).

CONCLUSIONLow oxygen reduced Kv1.5 mRNA and protein expressions, U0126 could resistant the Kv1.5 channel lower expression caused by hypoxia. Anisomycin had no significant effect on Kv1.5 channel expression under hypoxia, but the expression of Kv1.5 was still significantly lower than the normal oxygen group. These data suggest that hypoxia may cause hypoxic pulmonary hypertension by interfering ERK1/2 signaling pathway to inhibit Kv1.5

Animals ; Cell Hypoxia ; Hypertension, Pulmonary ; Kv1.5 Potassium Channel ; metabolism ; MAP Kinase Signaling System ; Mitogen-Activated Protein Kinase 1 ; metabolism ; Mitogen-Activated Protein Kinase 3 ; metabolism ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; metabolism ; Oxygen ; Pulmonary Artery ; cytology ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley

Objective To study the effect of single chain variable fragment (ScFv) antibody to EC3-4 fragment of desmoglein (Dsg) 3 in a mouse model of pemphigus vulgaris.Methods The ScFv an- tibody to EC3-4 fragment of Dsg-3 was injected subcutaneously into neonatal BALB/c mice at different time points;the mice were then evaluated clinically,histopathologically and by direct immunoflorescence exami- nation for the development of lesions.Results When injected alone,the ScFv antibody did not induce the appearance of key clinical features of pemphigus vulgaris.The antibody also did not prevent the develop- ment of pemphigus vulgaris features induced by sera of patients with pemphigus vulgaris,regardless of the time point of injection of ScFv antibody.Conclusion The ScFv antibody to EC3-4 fragment of Dsg-3 lacks pathogenicity in neonatal BALB/c mice,and also could not inhibit the development of lesions induced by sera from patients with pemphigus vulgaris.

Objective:To express and purify the TRIM5? chimaera[TRIM5? H(R328-332)] protein and to explore the interaction between the TRIM5? H(R328-332)and HIV-1gag. Methods:The plasmid pET28aTRIM5? H(R328-332) was transformed to E.coli BL21 (DE3) strain ,and the expression of TRIM5? H(R328-332) protein was induced by IPTG,purified with Ni2+ chromatography.The expression and purification of TRIM5? H(R328-332) were analyzed by SDS-PAGE and Western blot,and the interaction between TRIM5? H(R328-332) and HIV-1gag was detected by co-immunoprecipitation,His pull-down and ELISA. Results:The recombinant plasmid pET28aTRIM5? H(R328-332) was successfully expressed in E.coli. The results showed that the purified full length TRIM5? H(R328-332) interacted with HIV-1gag protein. Conclusion:The human TRIM5? chimaera was expressed successfully in vitro,and the study demonstrates that the human TRIM5? chimaera interacts with HIV-1 gag in vitro.

We have developed a sensitive and rapid lateral-flow immunoassay (LFIA) for WSSV,using colloidal gold as an indicator.The fusion protein,VP (19+28),was expressed in E.coli,purified and used to prepare polyclonal antibodies.The purified anti-VP (19+28) IgG were conjugated with colloidal gold.Unconjugated anti-VP (19+28) IgG and goat anti-rabbit IgG were immobilized on nitrocellulose membranes.After assembly,three groups (5 individual animals in each group) of shrimp samples were tested which included healthy,moribund and dead shrimps.For each group,three different tissues (body juices,gills and hepatopancreas) were tested at the same time.In parallel,all the samples were also analyzed using PCR for comparison.Out of 45 samples tested,30 were detected as positive while 15 were classified as negative.The results of LFIA correlate with those obtained by the PCR analysis,indicating that these two detection methods have the same efficacy in the limited number of samples tested in this preliminary study.