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

The ideal treatment and recovery of osteoarticular injury remain to be resolved. Small intestinal submucosa (SIS), a naturally-occurring decellularized extracellular matrix, has been recognized as an ideal scaffold for tissue engineering and widely used in repairing various tissues and organs. Nowadays its application has also been gradually increased in the field of orthopedics. We reviewed laboratorial studies and clinical trails about the application of SIS in bone and joint repair, aiming to evaluate its effects on the repair of bone, cartilage, meniscus, ligament and tendon. SIS has showed promising results in repairing bone, meniscus, ligament or tendon. However, additional studies will be required to further evaluate its effects on articular cartilage and tendon-bone healing. How to optimize SIS material,is also a focused problem concerned with making SIS a potential therapeutic option with high value for orthopedic tissue repair.
Animals ; Cell- and Tissue-Based Therapy ; Humans ; Intestinal Mucosa ; cytology ; Intestine, Small ; cytology ; Joint Diseases ; physiopathology ; surgery ; therapy ; Tissue Engineering ; instrumentation ; methods ; Tissue Scaffolds ; chemistry

Objective To determine the level and clinical significance of 5-hydroxytryptamine (5-HT) and intestinal fatty acid binding protein (iFABP) in patients of severe acute pancreatitis (SAP) with intestinal dysfunction. Methods The serum and urine in 42 cases of SAP with intestinal dysfunction were collected at day 1, 3 and 7 after admission, respectively. The numbers of bowel sounds were recorded. 20 health subjects were selected as the control. Results The numbers of bowel sounds in health subjects were 5.6 ± 2.3/min. The numbers of bowel sounds in SAP patients at day 1,3 and 7 were 2.3 ± 0.7/min, 1.7 ± 0.2/min and 3.1 ± 1.1/min, respectively;which were significantly lower than that in the control group (P < 0.01). The level of 5-HT in the control group was(86.7 ± 9.5)ng/ml, while the levels of 5-HT in the SAP patients at day 1, 3 and 7 were (112.0 ± 17.8) ng/ml, (130.5 ± 19.7) ng/ml, (107.9 ± 16.3) ng/ml. The level of urine iFABP in the control group was (90.5 ± 19.8) pg/ml, while the levels of urine iFABP in the SAP patients were (1250.2 ± 425.3) pg/ml, (1586.9 ± 523.4) pg/ml, (1154.6 ± 394.0) pg/ml. The levels of 5-HT and urine iFABP in the SAP patients all were significantly higher than those in the control group (P <0.01). The levels of 5-HT and urine iFABP in the SAP patients at day 3 were significantly higher than those in the day 1 and day 7, while there was no significant difference between the two groups. Conclusions Determination of serum 5-HT and urine iFABP in patients with early SAP could effectively evaluate the intestinal function, and it may have potential clinical significance.

Objective To investigate the effect of Da-Cheng-Qi decoction on enteric functional disturbance in rats with acute experimental pancreatitis and its potential mechanism. Methods Seventy-two SD rats were randomly divided into sham operation group, acute pancreatitis (AP) group and Da-Cheng-Qi decoction treated group with 24 each. The AP model was induced by retrograde injection of sodium taurocholate (0.1 ml/100 g) into bitiopancreatic duet. Before modeling, the rats in treatment group received Da-Cheng-Qi decoction (2 g/100 g), and the rats in other two groups received 0.9% NaCl solution. The rats were sacrificed at 3, 6 and 12 hours with 8 each, and the blood samples were taken for detecting the level of amylase and the contents of 5-hydroxytryptamine (5-HT) by enzyme-linked immunosorbent assay. The colonic tissues 10 cm apart from trans-ligament incluing jejunum, terminal ileum and sigmoid colon were collected for examining the expressions of 5-HT_3 and 5-HT_4mRNA and proteins by RT-PCR and Western blotting respectively. Results ① The levels of serum amylase and 5-HT in AP group and treatment group were significantly higher than those in sham operation group (P<0. 01), but they were lower in treatment group than in AP group at each time points (P<0. 05). The level of 5-HT was increased in the initial stage and then decreased gradually both in AP group and treatment group. ③ The expressions of 5-HT_3 and 5-HT_4 mRNA and proteins were significantly decreased in the jejunum, terminal ileum and sigmoid colon in AP group compared with sham operation group (P<0. 01). Whereas the expressions of 5-HT_3 mRNA and protein,but not 5-HT_4, were increased in the treatment group in comparison with AP group (P<0.05). Conclusions The level of 5-HT is significantly increased in acute pancreatitis, but its receptors (5-HT_3 and 5-HT_4) are decreased, which may induce enteric functional disturbance. The Da-Cheng-Qi decoction may improve enteric dynamic failure by increasing the expression of 5-HT_4 and may be a choice for treatment of acute pancreatitis with enteric dynamic failure