Streptococcus pneumoniae(S.pn) is an opportunity pathogenic bacteria,environmental factors play a key role in the pathogenicity of S.pn. It is important to study virulent gene in vivo. The S.pn suicide plasmid containing gfp reporter was constructed by fusing the genes pneumolysin and gfp,in which gfp is an excellent molecule probe in vivo. The plasmid was integrated to No.22 S.pn by homologous recombination. The recombinant S.pn was gained and evaluated in aspects of fluorescence excitation, biological character and physio-activity. The results showed it is efficient and available to report the expression of virulent genes in vivo and in vitro, which will provide a new easy method for evaluating and screening the virulent genes of S.pn in vivo.

AIM: To provide experimental evidence for gene therapy of thrombophilia disease, we constructed the eukaryotic expression plasmid with human thrombomodulin (hTM) gene and observed the alteration of hTM expression on the surface of human umbilical vein endothelial cells (HUVECs) with and without the reconstructive plasmid. METHODS: The whole expressive fragment of hTM gene was amplified by PCR from human genome. Both hTM gene and pcDNA3.1(+)/neo empty vector was digested by HindⅢ and EcoRⅠ. Two digested fragments were ligated into pcDNA3.1/hTM with T_4DNA ligase. After identifying, the reconstructive plasmid transfected into HUVECs using lipofectin. The hTM antigen on the HUVECs was detected by immunohistochemistry. RESULTS: The hTM reconstructive plasmid was confirmed by double endonuclease redigesting and sequencing. About 10% HUVECs were transfected by pcDNA3.1/hTM plasmid with lipofectin and the high-level hTM was detected on the transfected cells. CONCLUSION: We constructed the pcDNA3.1/hTM plasmid successfully, and it could be expressed on the HUVECs. [

To study the relation between drug release and the drug status within curcumin-loaded microsphere, SPG (shirasu porous glass) membrane emulsification was used to prepare the curcumin-PLGA (polylactic-co-glycolic acid) microspheres with three levels of drug loading respectively, and the in vitro release was studied with high-performance liquid chromatography (HPLC). The morphology of microspheres was observed with scanning electron microscopy (SEM), and the drug status was studied with X-ray diffraction (XRD), differential scanning calorimetry (DSC) and infrared analysis (IR). The drug loading of microspheres was (5.85 ± 0.21)%, (11.71 ± 0.39)%, (15.41 ± 0.40)%, respectively. No chemical connection was found between curcumin and PLGA. According to the results of XRD, curcumin dispersed in PLGA as amorphous form within the microspheres of the lowest drug loading, while (2.12 ± 0.64)% and (5.66 ± 0.07)% curcumin crystals was detected in the other two kinds of microspheres, respectively, indicating that the drug status was different within three kinds of microspheres. In the data analysis, we found that PLGA had a limited capacity of dissolving curcumin. When the drug loading exceeded the limit, the excess curcumin would exist in the form of crystals in microspheres independently. Meanwhile, this factor contributes to the difference in drug release behavior of the three groups of microspheres.
Calorimetry, Differential Scanning ; Curcumin ; chemistry ; Drug Liberation ; Lactic Acid ; Microscopy, Electron, Scanning ; Microspheres ; Polyglycolic Acid ; X-Ray Diffraction

Child, Preschool ; Female ; Hand, Foot and Mouth Disease ; complications ; diagnosis ; therapy ; Humans ; Infant ; Male ; Pulmonary Edema ; etiology ; Radiography, Thoracic

To study the relation between drug release and the drug status within curcumin-loaded microsphere, SPG (shirasu porous glass) membrane emulsification was used to prepare the curcumin-PLGA (polylactic-co-glycolic acid) microspheres with three levels of drug loading respectively, and the in vitro release was studied with high-performance liquid chromatography (HPLC). The morphology of microspheres was observed with scanning electron microscopy (SEM), and the drug status was studied with X-ray diffraction (XRD), differential scanning calorimetry (DSC) and infrared analysis (IR). The drug loading of microspheres was (5.85 ± 0.21)%, (11.71 ± 0.39)%, (15.41 ± 0.40)%, respectively. No chemical connection was found between curcumin and PLGA. According to the results of XRD, curcumin dispersed in PLGA as amorphous form within the microspheres of the lowest drug loading, while (2.12 ± 0.64)% and (5.66 ± 0.07)% curcumin crystals was detected in the other two kinds of microspheres, respectively, indicating that the drug status was different within three kinds of microspheres. In the data analysis, we found that PLGA had a limited capacity of dissolving curcumin. When the drug loading exceeded the limit, the excess curcumin would exist in the form of crystals in microspheres independently. Meanwhile, this factor contributes to the difference in drug release behavior of the three groups of microspheres.

Objective:To obtain the high expression of Herpes Simplex Virus Type 1(HSV-1)Glycoprotein D gene. Methods:The Herpes Simplex Virus Type 1(HSV-1)Glycoprotein D(gD1) gene fragment containing dominant antigen epitopes confirmed by computer analysis was cloned by PCR technical and inserted into plasmid vector pTrxA. Then the recombinant plasmid was transformed into Rosetta. The expressed product was analyze by SDS-PAGE. Results:930 bp gene fragment was amplified by PCR as anticipated. Nucleotide sequencing showed a 100 % homology with that of the published sequence in GenBank. The molecular weight of the expressed protein was about 48kDa, Western blotting indicated that the antigenicity of the protein was good. Conclusion:The plasmid pTrxA-gd1 was constructed and a high efficiency expression of the gd1 gene from Herpes Simplex Virus Type 1(HSV-1)strain was made. The expressed product shows a good antigenicity.

Objective To evaluate therapeutic effects and toxicity of advanced non-small cell lung cancer (NSCLC)treated by combining chemotherapy on ifosfamide(IFO)and vinorelbine(NVB).Methods 107 cases pa- tients with advanced NSCLC were enrolled.IFO was given in a dosage of 1.5g/m~2 on day 1 to 4.and NVB in a dosage of 25mg/m~2 on day 1 and 8.It was repeated every three or four weeks,up to two to four cycles.Results Two patients had complete response and 40 patients had partial response.The overall response rate was 47.7% ,the median survival time 10.3 months,1-year and 2-year survival rate was 42% and 12.3%,respectively.The main toxicity was bone marrow suppression.Conclusion The regimen is effective,sale and tolerable in advanced non- small cell lung cancer therapy.

Objective To evaluate the relationship between combined muhigene detection and response to chemotherapy and prognosis in epithelial ovarian carcinomas(EOCS).Methods A total of 80 ovarian tissue samples taken from the surgical specimens of patients with EOCS of our hospital in the last two decades who had received chemotherapy "after surgery were paraffin-embedded.The samples were divided into 2 groups,good prognosis group(patients who survived more than 2 years,n=46)and poor prognosis group(patients who survived less than 2 years,n=34).The expression levels of ToPo-Ⅱ,Ki-67,MGMT, PCNA,p27,p53,pl6,P-gp,LRP,GST-?,bcl-2,C-myc,Fas,bax,MSH2,MRP and BCRP were investigated by the combination of tissue arrays and immunohistoehemical streptavidin-biotin peroxidase(SP) method in all samples.Data were analysed with SPSS 12.0 for windows.Results There were statistically significant differences in the positive expression levels of P-gp,BCRP,MGMT,MSH2,p27 and p16(62%, 50% and 50% in poor prognosis group vs 33%,28% and 28% respectively,P

In this review, methods for testing in vitro release rate of sustained release preparations of traditional Chinese medicine (TCM) and in vivo-in vitro correlation were introduced. Studies indicated that a good correlation between in vivo and in vitro release can be obtained by establishing methods of in vitro release and it is important for the development of sustained release drug delivery system in TCM.
Administration, Oral ; Delayed-Action Preparations ; Drugs, Chinese Herbal ; administration & dosage ; Medicine, Chinese Traditional ; methods

OBJECTIVETo provide inspiration and ideas for clinical treatment of coronary heart disease (CHD) by data mining technology based frequency analysis and cluster analysis of medical records, prescriptions and herbs in treating CHD by distinguished veteran doctors of traditional Chinese Dedicine (TCM).

METHODSTotally 386 medical cases were retrieved from Wanfang Data, Chinese Scientific Journals Database (VIP medical information resources system, China National Knowledge Infrastructure (CNKI), and Typical Collections of Medical Cases by Contemporary Distinguished Veteran Doctors of Traditional Chinese Medicine. They input into database trimmed after unified standard. Medication laws of CHD by distinguished veteran doctors of TCM were analyzed using frequency analysis and cluster analysis, and so on.

RESULTSDistinguished veteran doctors of TCM frequently used top ten herbs in treatment of C D as Salvia miltiorrhiz , Ligusticum wallichii, Trichosanthes kirilowi, Pinellia ternat, Angelica sinensis, Poria coco stragalu , Panax ginseng, Allium macrostemon, and Radix Ophiopogonis. Cluster analysis summarized that there were 16 herb pairs commonly used, 7drug assemblies consisting of 3 herbs and 5 drug assemblies consisting of multiple herbs.

CONCLUSIONSDistinguished veteran doctors of TCM mainly used herbs assemblies capable for invigorating Pi to resolve phlegm, and promoting qi and activating blood circulation in treating CHD. Meanwhile, they concurrently used herbs combination of nourishing Xin and tranquilization, and regulating yin and yang.

China ; Cluster Analysis ; Coronary Artery Disease ; therapy ; Data Mining ; Databases, Factual ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Medicine, Chinese Traditional