[Objective] To develop an HPLC method for the determination of berberine hydrochloride in GuZheNing liniment.[Methods]The chromatographic system consisted of Dimonsil C18 column and mobile phase of acetonitrile—50mmol?L-1 sodium dihydrogen phosphate(adjusted PH to 2.5 with H3PO4)(25∶75).The flow rate was 1.0 mL?min-1,and the detective wavelength was at 348 nm.[Results]The calibration curve was linear in the range of 0.106～0.530 ?g(n=5).The method was proved to be repeatable with RSD 0.7%(n=6).4 batches of sample were analyzed and the Berberine Hydrochloride contents were 19.11,18.81,18.83,18.92mg/100mg,respectively.[Conclusion]The method is selective,simple and accurate,and could be used for the quality control of this preparation.

[Objective]To develop an HPLC method for the determination of berberine hydrochloride in GuZheNing liniment.[Methods]The chromatographic system consisted of Dimonsil C18 column and mobile phase of acetonitrile-50mmol?L-1 sodium dihydrogen phosphate(adjusted PH to 2.5 with H3PO4)(25:75).The flow rate was 1.0 mL?min-1,and the detective wavelength was at 348 nm.[Results]The calibration curve was linear in the range of 0.106～0.530 ?g(n=5).The method was proved to be repeatable with RSD 0.7%(n=6).4 batches of sample were analyzed and the Berberine Hydrochloride contents were 19.11,18.81,18.83,18.92mg/100mg,respectively.[Conclusion]The method is selective,simple and accurate,and could be used for the quality control of this preparation.

Objective To establish a method for efficient,accurate genotyping and nucleoside drug-resistant mutation analysis for hepatitis B virus ( HBV ).Methods The 48 HBV serum samples were collected from the Third People's Hospital of Taiyuan from July to August 2011,and HBV DNA were extracted using the commercial kit.The HBV whole genome and P gene were amplified and sequenced.Each HBV sample was genotyped by both constructing phylogenetic trees and genotyping software analysis.The results from two strategies were compared for every sample.Results A total of 48 HBV full genome sequences were identified into 12 B and 36 C genotype's by both constructing phylogenetic trees and genotyping software analysis,which was exactly the same as the analysis using P gene fragment sequencing.Seven forms of nucleoside drug-resistant mutation were found in the P gene for all the samples,with the ratio of 27.1％ ( 13/48 ),in which all the mutation forms were associated with lamivudine or adefovir,and no other nucleotide drugs-related resistance mutations existed.In addition,there were 11 B and 35 C genotype and 2 B/C hybrid type with the analysis using Real-time PCR genotyping for the 48 samples.Conclusion P gene sequencing can be used as a new clinical method for efficient,accurate HBV genotyping and resistant mutation analysis,which provides guidance for hepatitis B treatment.

Objective To study the expressions of CD24 and SLeX in lung cancer tissues,and evaluate the relation ship with development and progression of lung cancer.Methods Immunohistochemistry was used to detect the expressions of CD24 and SLeX in 70 specimens of lung cancer.Results The positive expression rates of CD24,SLeX in non-small cell lung cancer tissues were 41.3% and 63.5% respectively,There were no expression and weak expression respectively in small cell lung cancer (SCLC) tissues. The positive rates of CD24,SLeX in adenocarcinoma were 55.9% and 76.5%,respectively,which were higher than those in squamous cell carcinoma (24.1%,48.3%)(P0.05).In lung adenocarcinoma and squamous cell carcinoma,the positive expression rate of SLeX in stage Ⅲ (83.3%) was higher than that in stage Ⅰ-Ⅱ (55.6%),the positive rate in poor differentiated lung cancer(80.8%) was higher than those in moderate and well differentiate cancer(51.4%),the positive rate in female patients (76.7%) was higher than that in male patients (51.5%)(all P

Objective To investigate the effect and monitoring point of continuous veno-venous hemofil-tration (CVVH) in patients with multiple organ dysfunction syndrome (MODS). Methods In 22 patients with MODS,a double lumen catheter was put into the central vein,and the CWH was performed with the BRAUN Diapact CRRT. The level of BUN,Scr,serum potassium and arterial blood gas were measured 30 minutes before and after CVVH. The plasma TNF-α、IL-1、IL-8 were measured by ELISA. Vital signs were monitored dur-hag treatment process. Results The vital signs of all patients` was stable, the levels of BUN,Scr and serum potassium decreased significantly after CVVH. The plasma concentrations of TNF-α,IL-1 and IL-8 gradually decreased. Conclusions CWH can improve the blood biochemical markers,remove inflammatory cytokines in plasma,stsblize the vital signs during treatment,which is suitable for patients with MODS.

Objective To identify hepatitis C virus(HCV)-specific cytotoxicity T lymphocytes (CTL)epitopes by the combination of T epitopes prediction software and in vitro assays.Methods HCVspecific CTL epitopes were predicted by T epitope prediction software Rankpep and then candidate HCV-specific CTL epitopes were selected.Candidate HCV-specific CTL epitopes were used to stimulate PBMC of HCV-infected patients and healthy volunteers.and then enzyme-linked immunospot(ELISPOT)and intracellular cytokine staining(ICS)were used to measure the frequencies of IFN-γ-producing cells in total PBMC and the percentages of IFN-γ+CD8+T cells in total CD8+T cells,respectively.Results Five candidate CTL epitopes[NS3 450(TVPQDAVSR),NS3 594(GPTLLYRL),Ns4b 78(sMMAFSAAL),NS5a 416(SEENVSVVF)and NS5a 367(TVSSALAEL)]were used to stimulate PBMC of ten HCV-infected patients and two healthy volunteers.PBMC of seven HCV-infected patients secreted IFN-γ while PBMC of healthy volunteers did not produce IFN-γ.The frequencies of peptide-specific IFN-γ-producing cells ranged from 5 to 36 SFC/105 PBMC and the percentages of peptide-specific IFN-γ+CD8+T cells ranged from 0.02%-0.25%.Conclusion Resuhs of ELISPOT assay and ICS assay confirm that these five peptides NS3 450,NS3 594,NS4b 78,NS5a 416 and NS5a 367 are identified as Hovel HCV-specific CTL epitopes.

The aim of this research was to evaluate the influence of steam sterilization on poly(ethylene glycol-terephthalate) and poly(butylene terephthalate) copolymer (PEGT/PBT) and its vascular cells compatibility, which was used as the scaffolds in vascular tissue engineering. Endothelial cells, smooth muscle cells and fibroblasts were cultured separately on the films after steam sterilization and after ultraviolet sterilization. These cells can grow well on the films after ultraviolet sterilization, while they can hardly adhere on steam sterilized films. Differential scanning calorimetry, static contact angle, X-ray photoelectron spectroscopy, surface carboxyl density quantity, H-nuclear magnetic resonance and scanning electronic microscope were employed to characterize the properties of poly(ether-esters) films before and after sterilization. These results showed that steam sterilization had little effect on the surface morphology and on the constitution of the copolymer, but the copolymer segments were redistributed during steam sterilization. The hydrophilic poly(ethylene glycol) (PEG) and the end carboxyl groups transferred from the bulk and enriched on the surface and the degree of crystallinity of hard segments increased slightly. Both the end carboxyl and PEG enriched on the surface can hinder the protein adhesion on the surface; so, lacking in receptor, the vascular cells cannot adhere on the films surfaces.
Biocompatible Materials ; chemistry ; Cell Adhesion ; Endothelial Cells ; cytology ; Humans ; Polyesters ; chemistry ; Polyethylene Glycols ; chemistry ; Polyethylene Terephthalates ; chemistry ; Steam ; Sterilization ; Surface Properties ; Umbilical Veins ; cytology

Objective To develop a gene chip for the detection of pathogens causing genital ulcer diseases (GUDs). Methods Specific probes of 4 different pathogens were designed and synthesized. Gene chip was prepared by blotting the probes onto specially treated glass slides with the use of a robotics. Target genes of standard strains for the 4 different pathogens and the clinical specimens were amplified by PCR with Cy5 fluorescence labeled primers. The labeled amplicons were hybridized with gene chips, and then scanned and analyzed using computer software. Results The fluorescence signal for specific pathogen could be found in the gene chip, illustrating that one specific fluorescence signal denoted a single pathogen, and the combination of different signals denoted the corresponding co-existence of pathogens. Examination of 40 clinical specimens obtained from 40 patients with genital ulcers with gene chip was in good concordance with dark-field microscopy plus PCR or HSV culture plus PCR, showing Kappa values of 0.882 and 0.947, respectively. In addition, mixed infections were detected in 2 specimens. Conclusion Gene chip is a sensitive method with a reliable result and it can detect multiple infections simultaneously.

AIM: To establish a HPLC-UV method to determine sunitinib in rat plasma and mouse tissues, and to study its pharmacokinetics in rats and tissue distribution characteristics in mice. METHODS: The biotic samples were prepared by protein precipitation followed by a stereoselective analysis of sunitinib was achieved on Waters XBridgeTM C18 (4.6 mm×250 mm, 5 μm) with a mobile phase composing of methanol-0.02 mol/L sodium dihydrogen phosphate (70:30) at a flow rate of 1.0 mL/min. The detection wavelength was 310 nm, and the column temperature was 25 ℃. RESULTS: The calibration curve for rat plasma sunitinib was linear in the range of 0.019 2-15.34 μg/mL. The linear ranges in mice brain and kidney were 0.038 3-11.50 and 0.038 3-69.00 μg/mL, respectively. After intragastric administration of sunitinib at a dose of 20 mg/kg to rats, the pharmacokinetic characteristics were Tmax=9.0 h, Cmax=0.194 mg/L, t1/2=18.4 h, AUC(0-∞)=6.8 mg•L-1•h. And the absolute bioavailablity was 47.1%. It was indicated that sunitinib could permeate the blood brain barrier, but the concentration was lower in brain and higher in kidney. CONCLUSION: A HPLC-UV method for the determination of sunitinib in rat plasma and mouse tissues was established. The method is simple, rapid, reliable, and provides a reference for the clinical application of sunitinib.

This study was aimed to provide references for techniques in future application of traditional and natural drugs for tumor prevention and treatment. Domestic and foreign literatures on applications of gene chip technology in antitumor studies with natural and traditional medicine from 2000 to 2014 were reviewed. Corresponding database was established from aspects of published articles, drug intervention types, study fields, sample sources, chip technology platforms, repeatability of gene chip experiments, criteria of differential genes, and validation of gene chip experiments. Application experiences of gene chip in antitumor natural and traditional drugs were summarized. Shortcomings of gene chip technology application were analyzed deeply. The results showed that experimental gene screening was limited at the cellular level. More attentions should be paid to experiments at the animal and clinical levels. Scholars had paid more attention to expression level of mRNA and less attention to gene regulation level and epigenetic research of microRNA chip and DNA methylation chip. Gene chip experiments were lack of repeatability, which directly affected the evaluation results of difference gene and reduced reliability of gene screening. Screening results of genes should be verified not only at the mRNA level, but also increased at the protein level. It was concluded that gene chip was one of the most mature technologies to detect the level of gene expression, which was widely used in the research field of traditional and natural antitumor drug studies. Researchers should try to avoid deficiencies mentioned above in experiments related to gene chip technology.