Pluronic modified polyamidoamine (PAMAM) conjugate (PF127-PAMAM) was prepared and the inhibiting effect of MDR against MCF-7/ADR was investigated with doxorubicin (DOX) as model drug. 1H NMR and FTIR spectra showed that the conjugate was synthesized successfully. Element analysis accurately measured that 27.63% amino of per PAMAM was modified by pluronic (PAMAM : PF127, 1 : 35.37 mole ratio). PF127-PAMAM showed an increased size and a reduced zeta potential compared to PAMAM. PF127-PAMAM had lower hemolytic toxicity and cytotoxicity due to the reduced zeta potential and the protection of PF127. Each PF127-PAMAM molecular could load 19.58 DOX molecules, and the complex exhibited sustained and pH-sensitive release behavior. PF127-PAMAM/DOX exhibited weaker cytotoxicity than free DOX in MCF-7 cells; while the complex showed much stronger reverse effect of drug resistance in MCF-7/ADR cells, and resistance reversion index (RRI) was as high as 33.15.
Dendrimers ; pharmacology ; Doxorubicin ; pharmacology ; Humans ; MCF-7 Cells ; drug effects ; Poloxamer ; pharmacology

This study is to prepare the pantoprazole sodium enteric-coated tablet which is compacted by pellets. The enteric-coated pantoprazole sodium pellets were prepared by fluid bed coating technology. The pantoprazole sodium enteric-coated tablets were prepared by direct compression of the enteric-coated pellets and suitable excipients. In vitro dissolution method and scanning electron microscope method were used for the observation of the drug release behavior before and after compression of the pellets. The optimized formulation is: the coating level is 55%, the plasticizer content is 20%, the ratio of Eudragit L30D-55/NE30D is 8 : 2, enteric-coated pellets/excipients (MCC/PPVP/PEG 6000 = 2 : 1 : 1) is 5 : 5, the enteric-coated tablets release in artificial gastric fluid in 2 h is less than 10%, while in artificial intestinal fluid in 1 h is more than 85%. The release behavior of pantoprazole sodium enteric-coated pellets-type tablet is quite well. And it may be used in industrial production.
2-Pyridinylmethylsulfinylbenzimidazoles ; administration & dosage ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Drug Carriers ; Drug Compounding ; methods ; Excipients ; Microscopy, Electron, Scanning ; Plasticizers ; chemistry ; Polyethylene Glycols ; chemistry ; Polymethacrylic Acids ; chemistry ; Proton Pump Inhibitors ; administration & dosage ; chemistry ; Solubility ; Tablets, Enteric-Coated ; chemistry ; Technology, Pharmaceutical

PEG-modified magnetic Fe3O4 (Fe3O4-PEG) nanoparticles were sythesized using a solvothermal reaction and characterized with transmission electron microscopy (TEM) and thermo gravimetric analysis (TGA). The photothermal effect and photothermal destruction of cancer cells were evaluated. Then the doxorubicin loaded Fe3O4-PEG (DOX-Fe3O4-PEG) nanoparticles were prepared. The cytotoxicity and combined chemotherapy/photothermal therapy (PTT) effect were investigated. Uniform PEG coated Fe3O4 nanoparticles with particle size of 155 nm were obtained in the experiment. The loading and release of doxorubicin on Fe3O4-PEG were pH-dependent. The drug loading capacity in water was 21%. The results of MTT indicated a good biocompatiblity of Fe3O4-PEG nanoparticles and high cytotoxicity of DOX-Fe3O4-PEG. In combined therapy experiment, photothermal therapy demonstrated unambiguously enhanced chemotherapy efficacy. In conclusion, the obtained Fe3O4-PEG nanoparticles which exhibit good photothermal effect and drug loading capacity can be used for chemotherapy and photothermal therapy. The synergetic anti-tumor activity indicates the potential for the combined application of chemotherapy and photothermal therapy in cancer treatment.
Antibiotics, Antineoplastic ; administration & dosage ; pharmacology ; Cell Survival ; drug effects ; Doxorubicin ; administration & dosage ; pharmacology ; Drug Carriers ; Ferrosoferric Oxide ; chemistry ; Humans ; Hyperthermia, Induced ; MCF-7 Cells ; Magnetite Nanoparticles ; chemistry ; Particle Size ; Polyethylene Glycols ; chemistry

Objective: To evaluate analytical performance and clinical application value of a one-step HBV DNA quantitative detecting system. Methods: Analytical performance of the one-step HBV DNA quantitative detecting reagents included precision, residual contamination, accuracy, functional sensitivity and analytical measurement range were verified by collecting high concentration samples and external quality control samples from Jiangsu provincial clinical test center. Results: The within-run coefficient of variation (CV) of both low and high concentration samples were below 5%, meanwhile the intra-assay CV was below 3/5 TEa and inter-assay CV was below 4/5 TEa. There was no residual contamination and the analytic accuracy met the requirement of external quality assessment (EQA). Functional sensitivity was able to attain 100 IU/ml, while the day to day CV was below 20%. It exhibited a benign linear relation from 7.58×10¹ to 7.58×10⁸ IU/ml. Conclusions The analytic performance of a new testing system must be evaluated particularly before detecting samples of patients by quantitative tests. This study proves that the one-step HBV DNA quantitative detecting reagents can meet requirement of hepatitis B screening and clinical therapy monitoring, which is economic and simple for clinical routine tests.

BACKGROUNDLower fluence of 585-nm flashlamp-pumped pulsed dye laser has been successfully used as a nonablative technique in the treatment of wrinkles. The objective of this study was to evaluate the effect of the pulsed dye laser (585 nm) on the production of collagen and the mRNA expression of collagen related gene in fibroblasts in vitro.

METHODSCultured fibroblasts were treated with a 585-nm flashlamp-pumped pulsed dye laser (fluence 3 J/cm(2), 4 J/cm(2), spot size 7 mm, pulse duration 450 micros). The production of collagen and the mRNA expression of transforming growth factor (TGF)-beta1, SMAD2, SMAD3, SMAD4, SMAD7 and type I procollagen alpha1, alpha2 in fibroblasts were investigated by colorimetry or real time polymerase chain reaction.

RESULTSThe production of collagen was significantly up-regulated after treatment with a 585-nm flashlamp-pumped pulsed dye laser with a fluence of 3 J/cm(2) (P < 0.001). The mRNA expression of TGF-beta1, SMAD2, SMAD3, SMAD4, SMAD7 and procollagen I was significantly up-regulated after treatment with a 585-nm flashlamp-pumped pulsed dye laser with a fluence of 3 J/cm(2) (P < 0.001). No significant difference of mRNA expression of SMAD2, SMAD3, SMAD4, SMAD7 and type I procollagen was found between controls and fibroblasts treated with pulsed dye laser with a fluence of 4 J/cm(2) (P > 0.05).

CONCLUSIONSLower fluence (3 J/cm(2)) pulsed dye laser increased the collagen production in fibroblasts by up-regulating TGF-beta1, SMAD2, SMAD3, SMAD4, SMAD7 and type I procollagen mRNA expression. These may be the reason it can be effectively used in the treatment of wrinkles.

Analysis of Variance ; Cells, Cultured ; Collagen ; biosynthesis ; Fibroblasts ; cytology ; metabolism ; radiation effects ; Gene Expression ; radiation effects ; Humans ; Lasers ; Procollagen ; genetics ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Smad2 Protein ; genetics ; Smad3 Protein ; genetics ; Smad4 Protein ; genetics ; Smad7 Protein ; genetics ; Transforming Growth Factor beta ; genetics ; Transforming Growth Factor beta1

To express and prepare polyclonal antibody of Human papillomavirus type 6b (HPV6b) E7 protein. a prokaryotic expression vector pGEX-4T-2/HPV6b E7 was constructed and GST-HPV6b E7 fusion protein was expressed as a soluble protein in E. coli. The expressed fusion protein was purified via Glutathione-Sepharose 4B column and thrombin cleavage in order to obtain HPV6b E7 protein. Polyclonal IgG antibody was prepared by immunizing New-Zealand rabbits with HPV6b E7 protein. Western-Blot and immunofluorescence analysis showed that the polyclonal IgG antibody could specifically recognize HPV6b E7 protein and its titer was identified. SDS-PAGE analysis demonstrated that large amounts of soluble GST-HPV6b E7 fusion protein was expressed in E. coli after 3.0-6.0 hours of IPTG induction. Polyclonal IgG antibody successfully prepared from immunized rabbits showed high titer and high specificity as confirmed by Western-Blot and immunofluorescence. The preparation of anti-HPV6b E7 polyclonal antibody will facilitate further research on the biological and immunological functions of HPV6b E7 protein.
Animals ; Antibodies ; immunology ; Escherichia coli ; genetics ; metabolism ; Female ; Gene Expression Regulation, Bacterial ; HEK293 Cells ; Humans ; Immunoglobulin G ; immunology ; metabolism ; Oncogene Proteins, Viral ; genetics ; immunology ; metabolism ; Rabbits ; Recombinant Fusion Proteins ; genetics ; immunology ; metabolism

The study is to prepare taste masking and enteric-coated clarithromycin granules by melting and fluid bed coating technology. Clarithromycin and matrix materials were melted at a certain temperature, and then made into particles by fluidized bed coating. X-ray powder diffraction and scanning electron microscopy were used to identify the crystal and morphology of drug loading granules. In vitro dissolution method was used for the observation of the drug release behavior. The results showed that the drug particles size range was 0.2 - 0.6 mm; the crystal form of clarithromycin in the granule did not change; enteric-coated granules accumulated release in 0.1 mol L(-1) hydrochloric acid in 2 h was less than 10%, while in pH 6.8 phosphate buffer in 1 h was more than 80%. The taste masking and enteric-coated clarithromycin granules not only have good taste masking effect, but also have a good release behavior. It is expected to have better clinical application.
Clarithromycin ; administration & dosage ; chemistry ; Crystallization ; Drug Carriers ; Drug Compounding ; methods ; Excipients ; chemistry ; Microscopy, Electron, Scanning ; Particle Size ; Tablets, Enteric-Coated ; Taste ; Technology, Pharmaceutical ; methods ; X-Ray Diffraction

Objective To investigate the clinical significance in MRP1/CD_9 expression in cervical squamous cancer tissues and normal cervical tissues.Methods The expression of MRP1/CD_9 were assayed by SABC immunohistochemical methods in 53 cases of cervical cancer tissues and 13 cases of normal cervical tissues.Results Positive expression of MRP1/CD_9 was detected in 13 normal cervical tissue.MRP1/D_9 ex- pression is down-regulated in cervical carcinoma(P

OBJECTIVETo investigate the feasibility, safety and oncological effect of totally laparoscopic total gastrectomy (TLTG).

METHODSThe clinical data of TLTG cases and open total gastrectomy (OTG) patients between November 2007 and October 2011 were analyzed. Also compared the feasibility, safety and short-term outcomes of TLTG with OTG.

RESULTSNinty cases were analyzed. There were 18 cases in the TLTG group and 72 cases in the OTG group. Operation time was significantly longer in the TLTG group ((310 ± 86) minutes) than in the OTG group ((256 ± 57) min, t = 4.963, P = 0.002), However, the blood loss were significantly lower in the TLTG group ((136 ± 84) ml vs. (359 ± 141) ml, t = -11.734, P = 0.000). The post operative morbidity was similar between the TLTG and OTG group. First flatus time (t = -7.020), first diet time (t = -6.166 and -5.698), and post operative hospital stay (t = -4.610) were significantly shorter in the TLTG group than in the OTG group (P < 0.05).

CONCLUSIONSLTG is a safe and feasible procedure with quick post-operation recovery. The laparoscopic side-to-side esophagojejunal anastomosis is a safe and feasible method of alimentary reconstruction after laparoscopic total gastrectomy.

Adult ; Aged ; Female ; Gastrectomy ; methods ; Humans ; Laparoscopy ; Laparotomy ; Length of Stay ; Lymph Node Excision ; Male ; Middle Aged ; Stomach Neoplasms ; surgery ; Treatment Outcome

Based on the deletion information of high virulent PRRSV genome, 3 oligonucleotide primer were designed and synthesized. Specific and sensitive reverse transcription-PCR (RT-PCR) assays were de-veloped for the detection of high virulent PRRSV. The sensitivity and specificity of RT-PCR assays were evaluated, the results showing that the detection limit of the assay was found to be 0.265 pg of tissue total RNA, and the protocol have no cross-reaction with classical swine fever virus, porcine circovirus type 2,pseudorabies virus, streptococcus, haemophilus parasuis and Escherichia coli. Then 36 cell cultures, two PRRSV live vaccine strains and 184 clinical specimens from 52 farms were tested. Five PRRSV field iso-lates were the high virulent PRRSV; two PRRSV live vaccine strains from normal PRRSV, and 123 speci-mens from 42 farmer were positive (only 1 specimen was normal PRRSV). This RT-PCR method proved to be accurate differential diagnosis of the high virulent PRRSV and normal PRRSV with the characteristics of rapidity, sensitivity and specificity, and has a strong clinical relevance.