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.

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

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.

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

No abstract available.
Behcet Syndrome*

Objective To prepare recombinant human prothrombin-2 expressed in Pichia pastoris, and assay the enzymatic and clotting activities of prothrombin-2 activated by prothrombin activator ecarin.Methods Human prothrombin-2 gene and Echis carinatus ecarin gene were synthesized separately on the basis of the cDNA sequences published in GenBank.The gene of prothrombin-2 was cloned into the expression vector pPICZαA.The expression vector pPICZαA/prothrombin-2 was transformed into glycoengineered P.pastoris, and then prothrombin-2 engineered P.pastoris was screened.The expression products were induced by methanol, purified by two-step chromatography and identified by diges-tion by PNGase F and analysis of pepetide fingerprint.The ecarin gene was cloned into the expression vector pcDNA3.1. The expression vector pcDNA3.1/Ecarin was transformed into HEK 293T cells and the culture supernatant of HEK 293T/Ecarin was collected.The reaction product of HEK 293T/Ecarin cell culture supernatant and purified prothrombin-2 was analyzed by S-2238,which was the chromogenic substrate for thrombin.Fibrinogen was used to measure blood clotting time. Results The purified protein of P.pastoris expressed prothrombin-2 culture supernatant was 37 ×103 .The relative molecular mass(Mr) of the purified protein was reduced to 35 ×103, which was consistent with the theoretical Mr of prothrombin-2 molecular weight.The purified protein was proved to be prothrombin-2 by peptide fingerprint identification. The purified prothrombin-2 processed by HEK 293T/Ecarin culture supernatant could hydrolyze S-2238 to produce yellow pNA, and D405 of pNA increased with the volume of the processed prothrombin-2 that could promote the plasma coagulation.The blood clotting time was close to that of the thrombin kit.Conclusion Prothrombin-2 is prepared by P.pastoris and activated toα-thrombin by ecarin.This technique may replace the method of extraction of prothrombin from plasma and can be used for the treatment of war wounds or for future clinical research.

Objective To develop an interleukin-4(IL-4) antagonist named M5-IgG1Fc protein constructed by genetic engineering of antibody Fc fragment-cytokine mutein fusion protein which has a long half-life time in plasma.M5-IgG1 Fc protein binds to IL-4 receptor but cannot activate downstream signalling pathway , which provides a basis for drug develop-ment for allergic diseases .Methods The synthesized interleukin-4 mutant gene ( named M5 ) was cloned into the expres-sion vector pBV220 and transformed into E.coli DH5α.Chimeric gene M5-IgG1Fc obtained by overlap extension (SOE) method was transformed into glycoengineered Pichia pastoris GJK01 through expression vector pPICZαA .Then M5-IgGFc fusion protein was obtained by protein purification after being induced by methanol in 72 hours.The anti-IL-4 biologicial ac-tivity assay of M5 and M5-IgG1 Fc was performed with CTLL-2/IL-4R cells and detected with MTT colormetry .Finally,the half-life time of M5 and M5-IgG1 Fc protein in mice was compared by detecting the remaining amount in plasma with ELISA kit.Results The M5 protein expressed in E.coli and M5-IgG1 Fc fusion protein expressed in P.pastoris GJK01 both had IL-4 antagonistic bioactivity .The EC50 of both, which inhibited 5.6 ×10 -2 nmol/ml of IL-4, were 0.31 ±0.05 and 0.77 ± 0.03 nmol/ml,respectively.The maximum of M5 in plasma at 0.5 h was 5.8 ×10 -2 nmol/ml but the remaining amount was 2.8%of the maximum at 2 h.M5 protein could not be detected after administration at 8 h because of the detection line . The maximum of M5-IgG1 Fc fusion protein was 4.7 ×10 -2 nmol/ml,while fusion protein M5-IgG1 Fc decreased to 4.3%of its maximum at 120 h and could not be detected at 168 h.Conclusion M5 protein has IL-4 antagonistic bioactivity .M5-IgG1 Fc fusion protein expressed in glycoengineered P.pastoris GJK01 has IL-4 antagonistic bioactivity and long retention time in mice,which can be potentially used for treatment of allergic diseases .