Objective Broadcast storm caused by excessive datastream is dealt with by using VLAN (Virtual Local Area Network). Methods Through the VLAN, according to the location, roles and departments, network users or internet users, which application procedures and protocols are devided info groups. Results According to the hospital's network equipment and application situation, the local area network division of 16 VLAN is designed. Conclusion There are very good application prospects on improving the network manageability and providing better security.

Objective To understand the sanitary quality of cosmetics in Wuhan. Methods The sanitary quality of cosmetics including hair care cosmetics, skin care cosmetics, beauty cosmetics, perfumes and cosmetics for special use sampled from 23 cosmetics manufactures and 15 large-scale whole-salers and retail traders in Wuhan were examined during 1996～2001. Results The annual qualified rates of cosmetics were 93.49%～98.69% in 1996～2000. It was 95.65% for hair care cosmetics, 92.41% for skin care cosmetics, 98.13% for beauty cosmetics, 100% for purfumes and 95.56% for special use cosmetics respectively. They also were 93.62%～98.69% for microbiological indexes and 99.17%～100% for chemical indexes respectively. The total count of bacteria of skin care cosmetics seriously exceeded the related sanitary standard. Conclusion The main problem of the sanitary quality of cosmetics was the microbiological pollution.

2-(Dimethylamino)ethyl methacrylate and sodium monochloroacetate were employed to synthesize [2-(Methacryloyloxy)ethyl] dimethyl ammonium acetate (CBMA) functional monomer.CBMA was grafted on the surface of silica by surface initiated atom transfer radical polymerization (SI-ATRP) to obtain silica-CBMA hydrophilic interaction stationary phase.Three silica-CBMA stationary phases with different grafted density of CBMA monomer were synthesized in SI-ATRP progress by changing the concentration of CBMA under the same conditions.The ability to separate organic acid compounds of the synthesized silica-CBMA stationary phases was evaluated under different conditions, including effects of pH value, salt concentration and content of water of mobile phase on retention of solutes.The results showed that the stationary phases could effectively separate organic acid compounds in HILIC mode, which followed a mixed mode of chromatography of ion exchange and hydrophilic interaction.The retention of solutes decreased with the increases of salt concentration of mobile phase, which consistent with the characteristics of ion exchange;the pH value of mobile phase had significant influence on ionization of the stationary phase and solutes, i.e., the retention of solutes increased as the increasing of pH value of mobile phase.However, the retention of solutes decreased with the increasing of the content of water in mobile phase, which was the typical characteristic of HILIC.The method of hydrophilic interaction chromatography combined with silica-CBMA stationary phases could conveniently determinate the content of vitamin C and rutin in rutin tablets, providing a new method for the separation and determination of strong polar samples.

AIM: To quantify cyclooxygenase-2 (COX-2) mRNA in carcinoma of larynx and evaluate the correlation between the quantity of COX-2 mRNA and clinical staging or histological grade. METHODS: The expression of COX-2 mRNA in 30 cases of carcinoma of larynx tissue and adjacent non-cancerous tissues were evaluated by PCR, which includes a fluorescence dye , SYBR green Ⅰ, and the sequence specific primer. The GAPDH was used as control. RESULTS: The specificity of products was proved to be COX-2 and GAPDH by the analysis of the melting curve of the amplified products and agarose gel electrophoresis. The expression of COX-2 mRNA was detected in all cancerous tissues of 30 patients (100%), but only in 12 adjacent non-cancerous tissues of 30 patients (40%). The N_ COX value of carcinoma of larynx tissue and adjacent non-cancerous tissues was 16.54?13.27 and 9.24?6.91, respectively, and the expression levels of COX-2 mRNA elevated significantly in laryngeal squamous cell carcinoma tissue and there were significant correlation between the expression levels of COX-2 mRNA and clinical stage or histological grade. CONCLUSION: The expression of COX-2 mRNA in carcinoma of larynx can be determined by real-time PCR technique. An increase in COX-2 mRNA may be associated with carcinogenesis of carcinoma of larynx, and it may be useful as a biomarker in laryngeal cancer.

AIM: To quantify cyclooxygenase - 2 (COX - 2) mRNA in carcinoma of larynx and evaluate the correlation between the quantity of COX - 2 mRNA and clinical staging or histological grade. METHODS: The expression of COX - 2mRNA in 30 cases of carcinoma of larynx tissue and adjacent non - cancerous tissues were evaluated by PCR, which includes a fluorescence dye , SYBR green Ⅰ , and the sequence specific primer. The GAPDH was used as control. RESULTS: The specificity of products was proved to be COX - 2 and GAPDH by the analysis of the melting curve of the amplified products and agarose gel electrophoresis. The expression of COX - 2 mRNA was detected in all cancerous tissues of 30 patients (100%), but only in 12 adjacent non - cancerous tissues of 30 patients (40%). The NCOX value of carcinoma of larynx tissue and adjacent non - cancerous tissues was 16.54 ± 13.27 and 9.24 ± 6.91, respectively, and the expression levels of COX- 2 mRNA elevated significantly in laryngeal squamous cell carcinoma tissue and there were significant correlation between the expression levels of COX - 2mRNA and clinical stage or histological grade. CONCLUSION: The expression of COX - 2 mRNA in carcinoma of larynx can be determined by real - time PCR technique. An increase in COX - 2 mRNA may be associated with carcinogenesis of carcinoma of larynx, and it may be useful as a biomarker in laryngeal cancer.

Acoustic Stimulation ; Animals ; Auditory Perception ; physiology ; Behavior, Animal ; physiology ; Noise ; adverse effects ; Rabbits ; Sound ; Swine

The inhibitory effects of two kinds of selective cyclooxygenase-2 inhibitors on the proliferation of human carcinoma of larynx Hep-2 in vitro and their corresponding mechanisms were investigated. Hep-2 cells were cultured with two kinds of selective cyclooxygenase-2 inhibitors (Sc-58125 and Celecoxib) at various concentrations for 24 h. Morphological changes were observed under the phase microscopy and the growth suppression was detected by using MTT colorimetric assay. Apoptotic DNA fragments were observed by agarose gel electrophoresis, and the cell cycle and apoptotic rate were detected by flow cytometry (FCM) respectively. Hep-2 cells became rounded and detached from the culture dish after being treated with Celecoxib for 24 h, however, they remained morphologically unchanged with Sc-58125. Sc-58125 could increase G2 phase cells, whereas, Celecoxib rose G1 phase cells. Both of the two effects were dose-dependent. Moreover, the Hep-2 cells cultured with 50 micromol/L and 100 micromol/L Celecoxib showed obvious apoptosis, with the nuclear DNA of cells exhibiting characteristic DNA ladder. So Sc-58125 could inhibit the proliferation of Hep-2 cells by altering the G2 phase cells. However, Celecoxib had the same effect by changing the G1 phase cells and inducing apoptosis at higher concentration.
Antineoplastic Agents/pharmacology ; Apoptosis/drug effects ; Cell Proliferation/drug effects ; Cyclooxygenase 2 Inhibitors/*pharmacology ; Dose-Response Relationship, Drug ; Laryngeal Neoplasms/*pathology ; Pyrazoles/*pharmacology ; Sulfonamides/*pharmacology ; Tumor Cells, Cultured

Objective: To investigate the therapeutic effects of ets-1 antisense oligoxydeonucleotide(AsODN) on gastric carcinoma. Methods: Cultured SGC-7901 cells were devided into control group, sense oligoxydeonucleotide(sODN) group and AsODN group. After transfection for 24 h, expression of ets-1mRNA was detected by RT-PCR, growth inhibition was detected by colone formation assay, in vitro invasive ability assay was carried out in Transwell chamber,the animal model of xenotransplanted gastric carcinoma in nude mice was established to detect invasive ability in vivo. Results: ets-1 AsODN could under-regulate the expression of ets-1 mRNA, number of colone formation of AsODN group was significantly lower than the other two groups(24.2?4.8 vs 47.6?8.1 vs 44.3?7.6, P

Objective To express and purify mammalian glycosylation modified trimeric Ebola virus trimeric glycoprotein (EBOV-GP) using novel glycoengineered Pichia pastoris.Methods The EBOV-GP and EBOV-GPΔMLDΔTM genes were cloned into the pPICZ-αA vector,electrochemically converted to glycoengineered Pichia pastoris,and compared with EBOV-GP expressed in HEK-293T cells.Glycosylation was analyzed by PNGaseF and EndoH digestion,while the target protein was purified by affinity chromatography and ion exchange chromatography.N-terminal sequencing was used to determine whether the signal peptide was correctly cleaved during protein translation and gel column analysis was used to find out whether the trimeric structure was formed.Results The results of PNGaseF showed that EBOV-GP expressed by glycoengineered Pichia pastoris and HEK-293T cells had the same relative molecular mass and N-glycosylation degree.EndoH digestion showed that the N-glycosylation modification of EBOV-GPΔMLDΔTM was in a non-high mannose form.N-terminal sequencing showed that the signal peptide of the GP protein itself was correctly excised.Gel column analysis showed that the purified protein was in a trimeric form.Conclusion An EBOV-GP is obtained with complex glycosylation modification based on Glycoengineered Pichia pastoris.

Objective:To construct a human Fab fragment phage display library and provide a platform for human antibody preparation.Methods:Peripheral blood lymphocytes were collected from healthy donor.The heavy chain Fd fragment and light chain of human immunoglobulin's genes were amplified by RT-PCR,and then cloned into phagemid pComb3XSS to generate human phage antibody library.Cutting with endonucleases such as SacⅠ,XbaⅠ,XhoⅠand SpeⅠto identify the insertion of the light chain or heavy chain Fd genes.IL-2 and digoxin as the antigen was used to scan the phage antibody library.Results:A phage antibody library of Fab had 8.4?107 members and it's recombinant rate was 70%.Through DNA sequencing of one positive clone,it was showed that its heavy chain belonged to IgG subvariety and its light chain to ? family.Conclusion:The success of constructing a nave human phage antibody library proves the useful of phage display system in human antibody preparation,and it can be used to select,purify and express of amylin Fab antibody.