OBJECTIVE:To prepare nattokinase immunoliposomes and its lyophilized product and to evaluate its pharmaceutical quality. METHODS: Nattokinse liposomes were prepared by film dispersion method, then the immunoliposomes were prepared by carbodiimide method coupled anti-human fibrin D-dimer monoclone antibody SZ-63 and lyophilized and evaluated about its pharmaceutical quality. RESULTS: Lyophilized immunoliposomes were multilamellar vesicles with its mean diameter, Zeta potentials at 251.6 nm, —34.6 mV, respectively. The lyophilized product of nattokinase immunoliposomes exhibited good appearance and satisfactory redispersibility, with its loss on drying at 14.30% and critical relative humidity at 45.66%. CONCLUSION: Nattokinase immunoliposomes and its lyophilized product have been prepared successfully.

Actins ; metabolism ; Adenoma, Pleomorphic ; metabolism ; pathology ; surgery ; Diagnosis, Differential ; Glial Fibrillary Acidic Protein ; metabolism ; Humans ; Infant ; Keratins ; metabolism ; Male ; Nasal Cavity ; pathology ; Neoplasm Recurrence, Local ; Nose Neoplasms ; metabolism ; pathology ; surgery ; Reoperation ; Vimentin ; metabolism

OBJECTIVE:To study the difference between the hydrophilic and hydrophobic fluoroquinolones(FQNLs)in?ducing resistant Klebsiella pneumoniae(Kpn)in vitro.METHODS:Hydrophilic ciprofloxacin(CIP)and hydrophobic sparfloxacin(SPFX)were used to induce FQNLs resistant mutants from a clinical susceptible isolate Kpn909with stepwise selection method,respectively,The susceptivity of bacteria to was determined by the standard method of two fold agar dilu?tion.RESULTS:The resistance of Kpn909to FQNLs was raised up gradually as FQNLs'concentration increased;Minimum inhibitory concentrations(MICs)of Kpn9098Selected by CIP at2?MIC(0.5mg/L)were16～32times higher than that of Kpn909,and showed resistant to ofloxacin(OFX),lomefloxacin(LMFX)and enofloxacin(ENX)with a MIC of8mg/L,16mg/L and8mg/L.Otherwise,SPFX-selected Kpn with a similar resistant level occurred at the concentration of8?MIC(1mg/L).The maximal selecting concentration of CIP and SPFX were128mg/L and64mg/L,and MIC of5FQNLs to the resistant mutants obtained at which were≥256mg/L and32～128mg/L,respectively.CONCLUSION:Both hydrophilic CIP and hydrophobic SPFX could induce Kpn the cross resistance to others FQNLs,but hydrophobic FQNLs may be superior to hydrophilic FQNLs to delay the resistance occurrence in Kpn.

ObjectiveTo evaluate biomechanical properties of transoralpharyngeal atlantoaxial reduction plate (TARP) prepared from magnesium alloy and titanium alloy for the atlantoaxial dislocation by using three-dimensional finite element analysis and to exam the feasibility of using magnesium alloy for preparation of TARP system so as to provide a theoretical basis for clinical surgery.MethodsA patient with typical atlantoaxial fracture dislocation was involved in the study,and received thin CT scan with clinically used titanium alloy TARP system for obtaining DICOM image data.Three-dimensional finite element analysis software was imported to simulate magnesium alloy and titanium alloy TARP systems for reduction and fixation.Then,stress changes of the atlas,axis,internal fixators and C2/3 zygapophysial joints were determined with three-dimensional finite element analysis and analyzed statistically.Results ( 1 ) The finite element model of atlantoaxial dislocation reduction and fixation had lifelike outline and good geometric similarity.There were 53 586 nodes and 180 784 units.(2) During the simulation of head in neutral position,the stress concentration region was C2/3 zygapophysial joints followed by the anterior arch,posterior arch and lateral mass of atlas respectively,and C2 vertebral arch again.( 3 )Magnesium alloy and titanium alloy TARP systems showed significant difference in stress distribution (P ＜0.05).Conclusions(1)The atlantoaxial model established according to its structure information on CT can be used for biomechanical experiments.(2) For the treatment of atlantoaxial dislocation using the existing titanium TARP system,maintaining the integrity of anterior and posterior arch of atlas and confirming the bone fusion in lateral mass can better keep the stability of the atlantoaxis.After atlantoaxial fusion,the increased stress of the zygapophysial joints of the adjacent segments accelerates structural degeneration,which should be closely followed up.( 3 ) Magnesium alloy TARP system for fixation and reduction shows the fall in peak value of the stress concentration region,and improvement of the uniformity of stress distribution as compared with titanium alloy TARP.

Objective To survey the antimicrobial resistance of nosocomial Klebsiella Pneumoniae (Kpn) isolates. Methods All the 68 Kpn strains were isolated and collected from nosocomial infections during April 2002 to March 2003. The susceptivity of bacteria and the production of extended spectrum ?-lactamases (ESBLs) were determined by Kirby-Bauer disc diffusion test and phenotypic confirmatory testing, respectively. Results All 68 Kpn strains were 100% susceptible to Menopenem and Imipenem but resistant to 14 other antimicrobials to different degrees. The first five drugs of high resistant rate were ampicillin (100%), amoxycillin/clavulanic acid (79.4%), sulphamethoxazole/trimothoprim (67.6%), cefotaxime (58.8%), gentamycin (47.0%). There were 28 isolates producing ESBLs (41.2%), which showed much higher resistance to ?-lactam, aminoglycosides, quinolones and sulphamethoxazole/trimothoprim than the non-ESBLs-producing strains. Conclusion Nosocomial Kpn isolates were most multidrug resistant, of which ESBLs-producing strains showed a higher resistance and which may involve other resistant mechanisms.

Objective To study the expressions of HSP70 and AQP4 in the H.Pylori-associated gastritis mice of Pi-Wei hygropyrexia syndrome;To investigate the mechanism of Mieyou Soup. Methods The mice were randomly divided into high concentration of Mieyou Soup group, low concentration of Mieyou Soup group, gastric triad group, model group and control group. BALB/c mice of H.Pylori-associated gastritis mice of Pi-Wei hygropyrexia syndrome model was established by composite factor. After modeling, the mice were administered for of continuous 14 days. The drug dosage of high concentration of Mieyou Soup group, low concentrations of Mieyou Soup group, and gastric triad group were 12.4, 6.2 g/kg, and 0.279 8 mg/kg, respectively. Western Blot was used to detect the expression of HSP70 protein, and immunohistochemistry was employed to detect the expression of AQP4 protein. Results Compared with control group, the expression of HSP70 and AQP4 in model group significantly increased (P<0.05);compared with the model group, the HSP70 in high concentration of Mieyou Soup group, low concentrations of Mieyou Soup group, gastric triad group increased, the AQP4 in high concentration of Mieyou Soup group, low concentrations of Mieyou Soup group, gastric triad group decreased, with statistical significance (P<0.01). Conclusion Mieyou Soup may play a role in treatment of H.Pylori-associated gastritis mice of Pi-Wei hygropyrexia syndrome by raising the expression of HSP70, and reducing the expression of AQP4 protein expression.

Objective To investigate the mechanisms of Mieyou Decoction in the treatment of Hp-associated gastritis of spleen-stomach damp-heat syndrome. Methods Seventy BALB/c mice were randomly divided into control group, model group, high concentration of Mieyou Decoction group, low concentrations of Mieyou Decoction group, and gastric triad group, with 14 rats in each group. BALB/c mice of Hp-associated gastritis of spleen-stomach damp-heat syndrome models were established by composite factor. After the success of modeling and continuous 14-day administration, the expressions of TLR2, TLR4 protein, TLR2 mRNA, and TLR4 mRNA were detected by immunohistochemistry and qPCR. ELISA was used to detect the expressions of IL-6 and IL-8 in the serum. Results Compared with control group, the expressions of TLR2, TLR4 protein, mRNA, IL-6, and IL-8 in model group significantly increased (P<0.01);the expressions of TLR2, TLR4 protein, mRNA, IL-6, and IL-8 in high concentration of Mieyou Decoction group, low concentrations of Mieyou Decoction group, gastric triad group were lower than model group, with statistical significance (P<0.01);the expressions of TLR2, TLR4 protein, mRNA, IL-6, and IL-8 in high concentration of Mieyou Decoction group were higher than gastric triad group, but the differences were not statistically significant (P>0.05). Conclusion Mieyou Decoction may play a role in the treatment of Hp-associated gastritis of spleen-stomach damp-heat syndrome through intervention in the expressions of TLR2 and TLR4.

Purpose To investigate whether FoxM1 participate in inhibitory effect of cisplatin (CDP) in resistant osteosarcoma cell lines by down-regulation of Rad51.Methods The resistant osteosarcoma cell lines were induced by gradually increasing dose intermittent action,and were named MG-63/R and HOS-MNNG/R respectively.The mRNA and protein level of FoxMl and Rad51 were detected by qRT-PCR and Western blot analysis in resistant cells and parental cells.The mRNA and protein level of FoxM1 and Rad51 were detected by qRT-PCR and Western blot analysis in resistant cells after treatment of 4 μmol/L Thiostrepton.The effect of single or combined treated of 4 tμmol/L Thiostrepton or 2 tμg/mL CDP on the rate of cell proliferation in resistant cells was examed by cell counting.Results Resistant osteosarcoma cell lines MG-63/R and HOSMNNG/R were established and stablely growthed in the concentration of 2 μg/mL CDP,and the resistance index was 30.52 and 37.87 respectively (severe CDP resistance).The mRNA and protein level of FoxM1 and Rad51 were significantly increaced in resistant cells compared with parental cells.The proliferation rate of resistant cells in conbination of 4 μmol/L Thiostrepton and 2 μg/mL CDP treated group was significantly lower than these two drugs single treated group.The level of mRNA and protein of FoxM1 and Rad51 were significantly decreased after 4 μmol/L Thiostrepton treatment in CDP resistant cells.Conclusion The results suggest that FoxM1 and Rad51 may participate in the resistant osteosarcoma cells to CDP.FoxM1 inhibitor Thiostrepton may strengthen the inhibitory effect of CDP in the resistant cells by down-regulation of Rad51.

This study examined the role of PI3K/Akt pathway in radiosensitization of DNA damage of cervical carcinoma cells. The 50% inhibition concentration (IC(50)) of cisplatin and docetaxel in HeLa cells was detected by Mono-nuclear cell direct cytotoxicity assay (MTT) in vitro. HeLa cells were treated by cisplatin/docetaxel of 10 percent of IC(20) alone or combined with LY294002 for 24 h, and then radiated by different doses of X-ray. The cell survival ratio was obtained by means of clone formation. One-hit multi-target model was fitted to the cell survival curve to calculate dose quasithreshold (Dq), mean lethal dose (D(0)), 2Gy survival fraction (SF(2)) and sensitization enhancement ratio (SER). The pAkt and total Akt expression was detected by Western blotting and DNA damage by neutro-comet electrophoresis. The HeLa cells were randomly divided into 7 groups in terms of different treatments: Control; radiation treatment (RT) group; LY294002+RT group; cisplatin+RT group; docetaxel+RT group; LY294002+cisplatin+RT group; LY294002+docetaxel+RT group. The apoptosis ratio of each group was measured by flow cytometry. The results showed that docetaxel and cisplatin significantly enhanced the phosphorylation of Akt in radiation-treated HeLa cells. The Dq, D(0) and SF2 in LY294002-contained groups were lower than those in docetaxel or cisplatin+RT group. The SER in the LY294002+docetaxel+RT group was 1.35 times that of the docetaxel+RT group, and that in the LY294002+cisplatin+RT group 1.26 times that of the cisplatin+RT group. The Comet electrophoresis showed that tail distance in the LY294002+cisplatin+RT group or LY294002+docetaxel+ RT group was longer than in the cisplatin+RT group or docetaxel+RT group. The apoptosis ratio in the LY294002+cisplatin+RT group or LY294002+docetaxel +RT group was higher than in the cisplatin+RT group or docetaxel+RT group. It was concluded that inhibiting PI3K/Akt pathway can increase the effect of docetaxel and cisplatin on the radiosensitivity of HeLa cells and DNA damage resulted from radiation.

The new intein-mediated PHB purify protein system is a high expression, automatic cutting, for purification, low-cost protein purification system,it is conducive to large-scale protein purification.Choose human antibacterial peptide LL-37 as the purification objects,which is poison to prokaryotic cell.We construct intein-mediated PHB purified human antimicrobial peptide LL-37 system through genetic engineering technology and use this system to purify LL-37. The results show that this system can highly express LL-37 fusion protein and purifiy the product as same size with expectations.