OBJECTIVE:To apply the biosensor technology to screening of the best method to extract and separate anti-endotoxin effective materials in Radix Paeoniae rubra.METHODS:The affinities of samples extracted by three different methods binding to lipopolysaccride(LPS)were determined with biosensor technology.Extracted materials were incubated with0.25EU/ml endotoxin for determination of change of binding activity.RESULTS:Material extracted by water showed high binding capa?bility with LPS,after incubation with endotoxin,still had highly effective anti-endotoxin component;1∶40diluted water extract could neutralize78.1%endotoxin.Determination results by biosensor technology and traditional limulus reagent showed no dif?ference.CONCLUSION:Water extraction could obtain more anti-endotoxin effective materials.Compared with traditional methods,biosensor technology is a fast,highly effective,direct and accurate method.

OBJECTIVE: To determine the content of tobramycin in Tobramycin Dexamethasone Eye Drops by HPLC-ELSD.METHODS: The sample was separated on VP-ODS C18 column.The mobile phase was 1.0% trifluoroacetic acid-methanol(95∶5) at a flow rate of 1.0 mL?min-1.The drift tube temprature was 45 ℃;the pressure of nebulizing gas was 3.5 bar,and the detection wavelength was set at 254 nm.RESULTS: The liner range of tobramycin was 156.0～436.0 ?g?mL-1(r=0.999 7).The limit of detection was 0.523 ?g and the limit of quantification was 1.744 ?g.The average recovery was 99.57%(RSD=0.92%).CONCLUSION: The method is accurate,reliable,specific,and suitable for the quality control of Tobramycin Dexamethasone Eye Drops.

According to differentiating syndrome and dividing patter, 41 cases of Facial Chloasma by Auricular Pressure in Shen ( MA-AH), Fei ( MA-IC 1 ), Neifenmi ( MA-IC 3 ), Luanchao ( MA-AT), Mianjia (MA-L) and Xia' erbei(MA-IC), the total effective rate was 87.8%.

Objective To evaluate the endotoxin-neutralizing activity of modeling peptides from the limulus antilipopolysaccharide factor (MPLALFs, Ms) in vitro. Methods The endotoxin-binding activity of Ms was examined by biosensor technique and shown in values of Kon and Kd. The endotoxin-neutralizing effect was analyzed by limulus amebocyte lysate test. Results The biosensor technique results showed that the Kon values of M_0, M_1, M_2, M_3, M_4, M_6, M_8 and M_ 10 binding to LPS 055∶B5 were (840?5.716), (549?6.532), (842?6.530), (627?2.450), (996?5.716), (814?8.982), (556?1.633) and (635?2.449) arc second, of which M_4 and M_1 had the highest and lowest endotoxin-binding activity, respectively. The M_4 reacted to LPS with a Kd of 72.377 ?mol/L. The results obtained by the limulus amebocyte lysate test were the same with those from the biosensor technique. Conclusion M_4 has a potential good endotoxin-neutralizing effect in vitro.

Objective To study the effect of Bifidobacterium spent culture supernatant (SCS) on adhesion of Pseudomonas aeruginosa to intestinal epithelial cells. Methods The intestinal epithelial cells were cultured,then divided into three groups: control,Pseudomonas aeruginosa adhesion group,SCS and Pseudomonas aeruginosa adhesion group. The effects of SCS on cell viability,the number of adhering and invasive bacteria were evaluated with MTT assay and lysis-counting assay in 1,3,6 h. Results At 3 h after SCS treatment,the amount of Pseudomonas aeruginosa decreased significantly,and the number of adhering and invasive bacteria decreased by 71% and 87% respectively. Conclusion SCS could protect the intestinal epithelial cells by inhibiting the adhesion and invasion of Pseudomonas aeruginosa.

Objective To study the effects of early enteral feeding on the preservation of intestinal mucosal barrier in severely burned patients. Methods Twenty-two patients with severe burn were randomly divided into early enteral feeding group (EF) and delayed enteral feeding group (DF). The levels of serum endotoxin and TNF-? were dynamically detected in the patients of both groups, and two unmetabolized sugars (lactose and mannitol) were orally administered in these patients on 1d, 3d and 5d of postburn. The concentrations of lactose and mannitol in urinary and the L/M ratio were observed. Intestinal permeability was assessed by the L/M ratio. Results The levels of serum endotoxin and TNF-? in severely burned patients were significantly higher than in normal (P

Objective To investigate the effects and mechanism of silicone gel sheeting on hypertrophic scar. Methods Using human hypertrophic scar as a research object, the changes of fibroblast, collagen metabolism and cytokine expression were observed to study the effects of silicone gel sheeting on hypertrophic scar by clinical observation, histomorphology and immunohistochemistry. Results Compared with the control group, the hypertrophic scar became thin, soft and light markedly, and the expression of TGF ? 1, TGF ? (RI) and ? SM actin were significantly decreased in therapeutic group. Conclusion Silicone gel sheeting may have an influence on scar via inhibiting the expression of TGF ? 1, TGF ?(RI) and ? SM actin in fibroblasts.

Objective To investigate the genetic mutation of the norA gene and its promotor from the wild-type drug-resistance Staphylococeus aureus(S.aureus)strains. Methods A total of 10 antibiotic-resistant S.aureus strains were isolated and screened from the burn wound for the sequencing and analysis of the nora gene and its promoter. Results There isolated 87 S.aureus strains from the burn wound flora,which were completely sensitive to vacomycin,highly sensitive to Quinupristin and Nitrofurantoin,but highly resistant to the other antibiotics,even up to91.7% of MRSA.There found the same point mutation(G→A) located at 1 349 sites of the norA gene coding region in all the S.aureus strains,saying that the amino acid was changed from Gly(glycin)to Asp(agpartic acid) in 291 sites.The resetpine reverse test showed that the MICs value of three antibiotics was lowered at various degrees in all 10strains.Conclusion NorA gene mutation is one of the mechanisms for antibiotic-resistance of S.aureus.

Objective To investigate the protective effect of chloroquine on endotoxemia mice and its inhibition on the release of cytokines induced by LPS. Methods A total of 40 mice of Kunming species were randomly divided into four groups: LPS group received LPS at 10 mg/kg, chloroquine group received chloroquine at 20 mg/kg, LPS plus chloroquine group received chloroquine at 20 mg/kg first, then LPS at 10 mg/kg and control group received only 0.9%sodium chloride at 200 ?l/20 g. The mortality was observed within seven days after injection via caudal vein. ANA 1 cell lines were cultivated in vitro . After chloroquine was first added into the cells for 3 hours, the releases of TNF ? and IL 6 in the supernatants induced by different concentrations of LPS were measured. Results Chloroquine could decrease the death of mice due to endotoxin. Mortality dropped from 100% to 50% ( P

Objective To explore the mechanism of intestinal epithelial cell membrane injury due to Pseudomonas aeruginosa adhesion. Methods The intestinal epithelial cells were cultured and adhered with Pseudomonas aeruginosa . The changes in the viability of the cells and the activity of membranous PLA 2, the calcium content of cell, contents of phospholipid and membrane fluidity were observed. Results At 3rd h after the adhesion of Pseudomonas aeruginosa , the viability of the intestinal epithelial cells decreased significantly, but the activity of membranous PLA 2, the calcium content of cell increased significantly; the contents of phospholipid(PL), phosphatidylinositol(PI) and phosphatidylcholine(PC) in cell membrane decreased gradually, but the membrane fluorescence polarization and microviscosity of intestinal epithelial cell membrane increased significantly. Conclusion The activation of PLA 2 and the degradation of phospholipid due to the overloading of calcium in intestinal epithelial cells after the adhesion of Pseudomonas aeruginosa to intestinal epithelial cells might be the fundamental factors to result in the reduction of membrane fluidity.