The objective of this study was to construct an improved thioredoxin fusion protein expression system, and express the cathelicidin-derived peptide, Lf-CATH2. The improved fusion vector Lf-CATH2-pET32α(-TS) was successfully constructed by firstly deleting the thrombin site and S tag from the pET-32α vector, then inserting the Lf-CATH2 plus a thrombin site instead. Afterwards, Lf-CATH2 was expressed in Escherichia coli as fusion protein. After the cleavage by thrombin, Lf-CATH2 was released and subsequently separated using affinity chromatography. The antimicrobial activity of purified Lf-CATH2 was also examined. The improved expression vector significantly increased enzyme cleavage efficiency by 37%, and Lf-CATH2 could be expressed in high yield and maintain the biological activity. This novel thioredoxin fusion protein expression system enables a quick production of high-yield bioactive cationic peptides like cathelicidins.
Cathelicidins ; biosynthesis ; Chromatography, Affinity ; Escherichia coli ; Genetic Vectors ; Recombinant Fusion Proteins ; biosynthesis ; Thioredoxins ; genetics

OBJECTIVE:To observe clinical efficacy and safety of levofloxacin and azithromycin in the treatment of community acquired mycoplasma pneumonia (CAMP).METHODS:A total of 108 patients with CAMP were randomly divided into group A and group B (n=54).Group A received intravenous administration of levofloxacin (0.6 g,qd) and group B intravenous administration of azithromycinin (0.5 g,qd) for 7 days.The clinical efficacy and ADR of 2 groups were observed.RESULTS:The total effective rates of group A and group B were 83% and 56%,there was significant difference between 2 groups (P0.05).CONCLUSION:The clinical efficacy of levofloxacin in the treatment for CAMP is better than that of azithromycin.Levofloxacin and azithromycin have good safety.

OBJECTIVE:To observe the efficacy and safety of domestic cefepime in the treatment of patients with acute exacerbation of chronic obstructive pulmonary diseases (AECOPD) complicated with respiratory failure. METHODS:A total of ninety-six patients with AECOPD complicated with respiratory failure were randomly assigned to receive cefepime 2.0 g bid iv gtt (Group A,n=48) or cefoperazone/sulbactam (Group B,n=48) at a dose of 2.0 g bid iv gtt for 7～14 day. RESULTS:The effective rate was 82% for Group A versus 62% for Group B(P0.05). CONCLUSION:The efficacy of cefepime is higher than that of cefoperazone/sulbactam in the treatment of patients with AECOPD complicated with respiratory failure,but both had good safety.

OBJECTIVE:To evaluate the cough-relieving and sputum-eliminating efficacy and safety of Guaifenesin Pseudoephedrine Hydrochloride-Codeine Phosphate Oral Solution.METHODS:By a randomized control study,a total of 60 patients characterized by nonproductive cough andlittle sputum were randomly assigned to receive Guaifenesin Pseudoephedrine Hydrochloride-Codeine Phosphate Oral Solution(Trial Group)or Ambroxol Hydrochloride Oral Solution(Control Group)l0 mL tid for 5～7 d.RESULTS:Both groups had a significant improvement in symptoms including cough and expectoration after treatment(P0.05).CONCLUSION:Guaifenesin Pseudoephedrine Hydrochloride-Codeine Phosphate Oral Solution is more effective in relieving cough than Ambroxol Oral Solution,and it has certain effect in eliminating phlegm.In addition,it is safe and has little adverse reaction.

The study evaluates the lipolysis rate and extent of type Ⅲ lipid formulations using testosterone undecanoate as a model drug after digestion with in vitro lipolysis model, and studies the digestive regularity with optical microscope and electrical conductivity. The results showed that for testosterone undecanoate type Ⅲ lipid formulations with castor oil as oil phase and Transcutol HP as latent solvent, the lipolysis rate and extent were increased with the increase of oil phase proportion and were decreased with excessive proportion of surfactant, in which can see liquid crystal phase during lipolysis process. The lipolysis rate of type ⅢB lipid preparations with different surfactant were ordered as Labrasol > Tween 80 > Cremophor EL, but the rate of type ⅢA is different in quick digestion phase and slow digestion phase. The lipolysis extent of type Ⅲ lipid formulations with different surfactant were ordered as Cremophor EL > Tween 80 > Labrasol. These may be related to the digestive effect of pancreatic lipase on different surfactants. This study implied that the lipolysis rate and extent of type Ⅲ lipid formulations are greatly influenced by the proportion of oil phase and surfactant, and the surfactant structure. These factors will affect the in vivo digestion and should be taken into account when screening type Ⅲ lipid formulations.

To identify whether a highly repeated GT sequence from DCA1 promoter from Dunaliella salina,which have been proved to be a salt-inducible promoter in our previous study,would be a salt-inducible regulation element,different primers were designed to amplify 6 different-length fragments of DCA1 promoter from D.salina by PCR.After these fragments were respectively inserted into the HindⅢ-BamH I sites of the vector pU?GUS,serial expression vectors containing the gus gene were generated.D.salina cells transformed with these recombinant plasmids by electroporation were grown in liquid media containing different concentrations of sodium chloride respectively.GUS enzyme activity was measured histochemically and fluorometrically.The results revealed that 3 fragments containing GT repeated sequence drove the external gus gene expression and the expression pattern of the gus gene was regulated by the concentrations of sodium chloride.Additionally,the 2 fragments without tandem GT sequence drove the gus gene expression,but the expression pattern of the gus gene wasn't regulated by the concentration of sodium chloride;Also,the upstream fragment of the tandem GT sequence wasn't able to drive the gus gene expression.In conclusion,the highly repeated GT sequence from the DCA1 promoter plays an important role in the salt-inducible regulation of DCA1 promoter from D.salina and might be a novel salt-inducible element.

Aged ; Arthroplasty, Replacement, Knee ; adverse effects ; Female ; Humans ; Radiography ; Severe Acute Respiratory Syndrome ; diagnostic imaging ; etiology

Objective To investigate the effects of bone mesenchymal stem cells (BMSCs)transplantation on the neurological function recovery of injured spinal cord and the underlying mechanism.Methods Rats were subjected to contusive spinal cord injury by using NYU spinal cord contusive impactor system ( NYC lmpactor).Seven days after spinal cord injury,the transplantation of BMSCs ( BMSCs group) or injection of PBS ( PBS group) was performed around the epicenter of injured spinal cord in rats.Basso-Beattie-Bresnahan (BBB) score was used to evaluate the function of spinal cord.The cavity volume of the injured spinal cord was measured and the axons in the injury center of spinal cord were examined under transmission electron microscopy.The BMSCs of the green fluorescent protein (GFP)transgenic rats were used to trace the transplanted cells and the survivor of BMSCs in the injured spinal cord and their differentiation into neural cells were observed.A mini-channel implantation model was employed to further investigate the role of BMSCs transplantation on the axonal regeneration.Results The BMSCs group showed a higher BBB score and a smaller lesion volume as compared with the PBS group.Transmission electron microscopy examination displayed that the number of axons in the BMSCs group was far more than that in the PBS group.A great number of BMSCs-GFP were founded around the center of the injured spinal cord at 4 weeks after BMSCs transplantation.lmmunohistochemistry showed that the implanted BMSCs-GFP did not express the surface marker of neurons,astrocytes and oligodendrocytes.In the mini-channel implantation model,NF-positive nerve fibers grew into the BMSCs-seeded channel,while there were no nerve fibers in the channel without seeding of BMSCs.Conclusions The BMSCs transplantation for the injured spinal cord promotes its functional recovery,and the related mechanism is in correlation with BMSCs transplantation inducing axonal regeneration.

Objective To study the detection rate of early esophageal carcinoma using magnifying endoscopy,and to evaluate the relationship between the imaging patterns of microvasculature change and his- tological diagnosis.Methods Two hundred and fourteen patients with esophageal mucosa roughness,ero- sion,plaque,abnormal color and indentation in conventional endoscopy and 16 healthy volunteers were en- rolled.The magnifying endoscopy images were graded as four patterns by intraepithelial papillary capillary loop(IPCL)changes after iodine dyeing.The biopsies underwent pathologic evaluation.The comparison be- tween the imaging patterns of endoscopy and histological diagnosiswas was evaluated.Results 80.4%(90/ 112)esophagitis was type 2,and 85.7%(12/14)early esophageal carcinoma was type 3 and type 4.The difference was significant between early esophageal carcinoma and normal mueosa(?~2=27.32,P

Objective To evaluate the reliability of transpulmonary thermodilution technique,for the quantification of extravascular lung water.Method Eighteen dogs were randomly divided into 3 groups:normal group(n=6),ALIp(acute lung injury caused by pulmonary monary origin causes)group(n=6)and a ALIexp(acute lung injury caused by extra-pulmonary origin causes)(n=6).ALI_(exp)model was prepared by intravenous injection of injection of oleie acid.ALI_p model was made by inhalation of detergent.Extravascular lung water(EVLW)was measured by PiCCO monitor(PULSION,Germany)before and immediately after the intratracheal introduction of normal saline into the alveoh of lungs of normal and ALI dogs.Results The EVLW of the normal group was(254.5?13.0)mL.The volume of the normal between pre-and postinstillation(?EVLW) was approximate to the volume of normal saline instilled with a good correlation(r=0.986,P