Objective To study the structure of class 1 integrons in 90 strains of Pseudomonas aeruginosa isolated during two periods of 1992-1996 and 2003-2005,and to get information about the structure changing of class 1 integrons by comparing their structures in two different periods.Methods Routine PCR and long PCR were performed to amplify the class 1 integrons and the gene cassettes they carried, followed with sequencing and blast via GenBank.Results Thirteen out of 41 strians ioslated during the period of 1992-1996 were positive on class 1 intergrons.Long PCR showed that the class 1 integron was 1868 bp in length and contained 2 resistance genes averagely.Six types of resistance genes of qacEA1 (n=6), sull (n=14),aadA1 (n=2),aadB (n=1),PSE-1 (n=2) and tetA (n=1) were found in these integrons, which consisted of 5 patterns of resistance cassette arrangements.Nineteen strains were proved to carry class 1 integrons in 49 isolates from 2003-2005.The mean DNA sequence length of them was 3383 bp with 3.6 resistant genes in averagely,10 types of resistance genes,qacEA1 (n=18),sull (n=25),aadA1 (n=6), aadB (n=7),aacA4 (n=2),PSE-1 (n=3),VEB-1 (n=4),OXA10 (n=1),cm1 A (n=1) and tetA (n =2),were identified in these integrons,which were composed of 9 patterns of resistance cassette arrangements.Conclusion In terms of produce length and resistance cassettes carried in the integrons, greater complexity is found in the structure of class 1 integrons in strains isolated during 2003-2005 than those during 1992-1996.

Plant bioreactor called mocular farming has enormous potential to produce recombinant protein infinitely. Products expressed in plants has nature physico-chemical property and bioactivity. Plant bioreactor could be an safe, economic and convenient production system which has been widely applied in industries and agriculture, especially in the life science and pharmaceutical industry. The application of recombinant transgenic plant in the production of vaccines, antibodies and pharmaceutical proteins has become a hot point in the plant genetic engineering both at home and broad. However, there are some limiting factors of application such as yield, downstream processing and so on. The advantages and research progress for the mocular farming of pharmaceutical proteins recent three years was discussed, focusing on the existing problems and new strategies in this area.

OBJECTIVETo compare the effect of reinforcing Shen method (RSM) and activating blood method (ABM) in treating osteoarthritis (OA) at the molecular level.

METHODSThe physical and chemical characteristics of components from respective recipes of RSM and ABM, and network features of component-target interaction network were analyzed by computer simulation methods including chemical space, molecular docking, and biological network, etc.

RESULTSThe chemical components of RSM and ABM were scarcely scattered with larger overlapping. Among established networks, the distribution of network features was partially similar in RSM and ABM. The average target number correlated with each component was 1.86 in RSM and 2.11 in ABM respectively. Each average target number was respectively correlated with 4.46 compounds and 3.93 compounds, reflecting multi-component and multi-target actions.

CONCLUSIONComputer simulation could intuitively trace out similarities and differences of two different methods and their interaction with targets, which revealed that the compatibility of RSM and ABM could have broader protein targets and potential synergism at the molecular level.

Computer Simulation ; Drugs, Chinese Herbal ; administration & dosage ; therapeutic use ; Humans ; Osteoarthritis ; drug therapy ; Phytotherapy ; methods

OBJECTIVETo assess the clinical efficacy, safety and compliance of tianwang buxin decoction (TWBXD) combined with dormancy hygiene education (DHE) and TWBXD alone in treatment of sub-healthy insomnia patients of yin deficiency fire excess syndrome.

METHODSThe multi-centered, single blinded randomized clinical trial design was adopted. One hundred and one sub-healthy insomnia subjects of yin deficiency fire excess syndrome were randomly assigned to two groups. The 50 in the treatment group were treated by combined treatment with TWBXD and DHE, while the 51 in the control group were treated with TWBXD alone. The therapeutic efficacy, Pittsburgh sleep quality index (PSQI) score, clinical global impression-improvement (CGI) score, quality of life made by WHO (WHOQOL-BREF) score, and safety in the two groups were compared.

RESULTSThe effective rate in the treatment group was 68.08%, lower than that in the control group (75.00%), but the difference between them was statistically insignificant. The PSQI score in the treatment group were reduced from 12.00 +/- 2.25 to 7.55 +/- 2.91 (P < 0.01). It was reduced from 11.68 +/- 2.21 to 7.16 +/- 3.13 in the control group (P < 0.01). The improvement of CGI score and WHOQOL-BREF score was also shown in the two groups after treatment (P < 0.01). No significant difference was shown in each index between the two groups. There was no significant difference in CGI between two weeks after drug withdrawal and by the end of the therapeutic course in the same group (P > 0.05). There was no statistical significance in inter-group comparison (P > 0.05).

CONCLUSIONSignificant effect was achieved by TWBXD combined with DHE and by TWBXD alone. Their efficacies were equivalent, with high compliance and safety.

Aged ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Hygiene ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Patient Education as Topic ; Single-Blind Method ; Sleep Initiation and Maintenance Disorders ; therapy ; Treatment Outcome ; Yin Deficiency ; therapy

OBJECTIVETo explore the material basis of Caulis sinomenii (CS) in treating osteoarthritis (OA), and to give a pharmacodynamic illustration for the multi-targeting therapeutics of CS.

METHODSThe computational methods, consisting of molecular docking and biological network were carried out to search the database targeting twelve important OA related enzymes: ASAMTS4, ASAMTS5, MMP-1, MMP-3, MMP-13, MMP-8, MMP-2, COX-2, COX-1, IL-1beta, TNF-alpha, iNOS, and map the ligand-target interaction networks about molecules from CS and DrugBank. After that, an aggregate analysis was performed to analyze the mechanisms of compositions in CS.

RESULTSTotally 14 had good interaction in all molecules in database with two or more than two of the OA correlated enzymes, and 6 molecules had interaction with four or more enzymes. Moreover, both herb ligand-target interaction network and drug ligand-target interaction network were similar in the interaction profiles and network features, which revealed multi-drugs effects in CS.

CONCLUSIONSThere were a lot of multi-target molecules in CS, providing pharmacodynamic illustrations for the multi-target therapeutics of Chinese medicine. Meanwhile, they supplied certain reference and inspiration for finding out new drugs for OA therapy.

Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Molecular Docking Simulation ; Osteoarthritis ; drug therapy ; Phytotherapy ; Sinomenium

Objective To observe the effects of a motor relearning programme (MRP) combined with different early acupuncture interventions on muscle tension and motor function recovery after cerebral infarction.MethodsA total of 90 patients with cerebral infarction who met the inclusion criteria were divided into three groups at random:a YANGMING meridian acupuncture and MRP group ( group A),an anti-spasm acupuncture and MRP group ( group B),and an MRP group ( group C ).All of the patients in all three groups were treated with routine medication.The National Institute of Health stroke scale (NIHSS),the composite spasticity scale (CSS),Fugl-Meyer assessment (FMA),the Fugl-Meyer balance scale (FM-B) and the modified Barthel index (MBI) were used to measure performance before treatment and after 4 weeks of treatment.Another comparison was intra-group between before and after treatment. ResultsThere were significant differences in the assessment results in all of the groups after treatment compared with those before treatment.After treatment,group B was superior to group C only in terms of NIHSS scores.There was no significant NIHSS score difference between groups A and C.The FMA,CSS and MBI results revealed significant differences among all three groups,with the scores of group A consistently the highest.The average FMA score in group B was significantly higher than in group C but there was no statistically significant difference in FM-B scores among the three groups. ConclusionMRP therapy combined with early acupuncture intervention can improve motor function and muscular tension after cerebral infarction.Anti-spasm acupuncture can improve motor function and control muscular tension effectively at the same time,making it beneficial for MRP training.

Objective To investigate the extracellular polysaccharide distribution and components of Ureaplasma urealyticum (Uu) after biofilm having been developed in.Methods The standard serotype 3 and serotype 14 belong to biovar Parvo,and the standard serotype 4 and serotype 8 belong to biovar T960 were employed to form biofilrns in vitro.Scanning electron microscope and confocal laser scanning microscope were used to analysis the biofilms and extracellular polysaccharide.We used combination of two different labeled lectins,Canavalia ensiformis(FITC-ConA) and Erythrina cristagalli(ECA) which bind to specific polysaccharide residues to visualize extracellular polysaccharide in biofilms,and average uorescence intensity was evaluated Results All the strains can form the biofilmsin vitro.The biofilm was honeycomb-Like structures mainly,and extracellular polymeric substances accounts for majority of proportions.All the extracellular polysaccharide could be combined with FITC-ConA and ECA,and the total average fluorescence intensity of FITC-ConA was higher than ECA( P＜0.001 ).Conclusion Ureaplasma urealyticum biofilm is honeycomb-like structures mainly.The extracellular polysaccharide contains,galactose,and N-acetyl glucan residual,and the glucose,mannose residual are the main components.

Objective To investigate the expression changes of aspartic proteinase (cathepsin D) and cysteine proteinase (cathepsin K) in photoaged fibroblasts. Methods The senescence of human fibroblasts was induced via culture in the presence of 8-methoxypsralen (MOP) of 50 mg/L in darkness for 24 hours followed by irradiation with UVA of 80 kJ/m~2. Then, aged fibroblasts were confirmed by senescence-associated β-galactosidase (SA-β-gal) staining. Real-time RT-PCR and Western blot were carried out to detect the mRNA and protein expressions of cathepsin D and cathepsin K in photoaged and normal control fibroblasts, respectively. Results Western blot showed a significant difference between photoaged and control fibroblasts in the grey scale of cathepsin D and cathepsin K (3.25 ± 0.33 vs 14.18 ± 2.25, f = 30.61, P < 0.01; 2.39 ± 0.66 vs 29.38 ± 4.62, t = 12.63, P< 0.01). The △Ct values for cathepsin D and cathepsin K mRNA were 2.79 ± 0.17 and -0.92 ± 0.06, respectively, in photoaged fibroblasts, significantly lower than those in the control fibroblasts (4.54 ± 0.34, 2.57 ± 0.13, t = 20.78, 28.50, respectively, both P < 0.01). According to the value of 2~(-△△Ct), the expression of cathepsin D and cathepsin K mRNA decreased 0.24 ± 0.021 and 0.09 ± 0.005 folds, respectively, in photoaged fibroblasts compared with the control fibroblasts. Conclusion The expression of cathepsin D and cathepsin K is decreased in photoaged fibroblasts.

Objective To investigate the role of cathepsin G in photoaged fibroblasts. Methods Human fibroblasts were cultured and induced to premature senescence using UVA + MOP methods. Senescence-associated-β-galactosidase (SA-β-gal) stain was used to evaluate the positive rate of aged cells. The mRNA and protein expression of cathepsin G in photoaged fibroblasts were detected by real-time RT-PCR and Western blot techniques. Results Over 98 % induced cells presented a positive SA-β-gal straining. The expression of cathepsin G, detected by Western blot, was increased to (1. 70±0. 028) times of the control. And RT-PCR revealed that the synthesis of cathepsin G mRNA was also up-regulated to 1. 42±0. 09. Conclusion The results of our study demonstrates a significant correlation between photoaged fibroblasts and cathepsin G. The up-regulation of cathepsin G may play an important role in the damages of extracellular matrix and activation of MMPS in photoaged human skin.

Background: Intralesional excision with curettage is the standard method of giant cell tumor (GCT) treatment, but the ideal filling material after curettage remains controversial. The purpose of this study was to compare the oncological and functional outcomes which underwent cementation or bone grafting after GCT curettage around the knee. Methods: We reported 136 cases with GCTs in distal femur or proximal tibia who accepted curettage from five clinical centers during the last 15 years. All patients were divided into two groups according to filling materials. Recurrence?free survival proportions were used to evaluate oncological outcomes while the Musculoskeletal Tumor Society (MSTS) 93 functional score was used to evaluate functional outcomes. Other parameters including surgical complication, general condition, and radiological classification had been analyzed. The valid statisitical data was analyzed using SPSS 13.0 software. Results: After GCT curettage, 86 patients (63.2%) accepted bone grafting while 50 patients (36.8%) accepted cementation. There was no statistical difference in age, gender, tumor location, radiological classification, fixation, follow?up time, and MSTS 93 functional score between cementation group and bone grafting group. The recurrence?free survival proportions showed that the recurrence rate in bone grafting group was higher than it in cementation group (P = 0.034). Surgical complication was lower in cementation group than that in bone grafting group but without statistically significant difference (P = 0.141). Conclusions: Parameters including patients'age, gender, tumor location, and radiological classification did not affect surgeons'treatments in cavity filling after GCT curettage. Cementation should be recommended because of easy usage, the similar postoperative knee function with bone grafting, and the better local tumor control than bone grafting.