ObjectiveTo explore the value of simvastatin in improving endothelial function in the patients with acute coronary syndromes in shorter time.Methods60 patients with acute coronary syndrome(acute myocardial infarction and unstable angina/non-ST elevation myocardial infarction) were randomized to be treated with placebo(n=30) or simvastatin 20 mg daily(n=30) for 3～5 d.At the admission and endpoint,Brachial ultrasound was used to measure endothelium-dependent flow-mediated dilatation(FMD) and response to endothelium-independent nitroglycerin. ResultsFMD was unchanged with placebo,but increased with simvasatin,from(2.65±2.95)% to(4.19±2.59)%(P=0.027).Responses to nitroglycerin were similar during the time course of the study in the 2 groups.The improvement of FMD was not correlated with the level of TC(R2=0.081,P=0.37),LDL-C(R2=0.056,P=0.46) or HDL-C(R2=0.073,P=0.40).ConclusionSimvastatin initiated early after acute coronary syndromes rapidly improves endothelial function in short course.No correlation has been detected between the pharmacological effects of simvastatin with the fall in TC and LDL-C.

Cutaneous Fistula ; Digestive System Abnormalities ; therapy ; Endoscopy, Gastrointestinal ; methods ; Female ; Humans ; Infant ; Treatment Outcome

Cleistocalycis Operculati Cortex is the dry bark of Cleistocalyx operculatus. It is the raw material of Compound Hibiscuse which is external sterilization antipruritic drugs. The quality standard of Cleistocalycis Operculati Cortex in Guangdong Province "standard for the traditional Chinese medicine" (second volumes) only contains TLC identification. It is unable to effectively monitor and control the quality of Cleistocalycis Operculati Cortex. A reversed-phase HPLC method was established for the determination of 3, 3'-O-dimethylellagic acid from Cleistocalycis Operculati Cortex and the content was calculated by external standard method for the first time. Under the selected chromatographic conditions, the target components between peaks to achieve effective separation. 3,3'-O- dimethylellagic acid standard solution at the concentration of 1.00 - 25.0 mg x L(-1) showed a good linear relationship. The standard curve was Y = 77.33X + 7.904, r = 0.999 5. The average recovery was 101.0%, RSD was 1.3%. The HPLC method for the determination of 3,3'-O-dimethylellagic acid in Cleistocalycis Operculati Cortex is accurate and reliable. It can provide a strong technical support for monitoring the quality of Cleistocalycis Operculati Cortex.
Chromatography, High Pressure Liquid ; methods ; Chromatography, Reverse-Phase ; methods ; Drugs, Chinese Herbal ; analysis ; Plant Bark ; chemistry ; Syzygium ; chemistry

OBJECTIVETo observe the travel, divisions, and the lengths, diameters, branches, artery supplies of the main segments of maxillary nerve.

METHODSFifty formalin-preserved adult half-head specimens with intravascular injection of red color emulsion were used for the gross and microanatomical studies of maxillary nerve. The lengths, diameters, branches and artery supplies of four main segments of maxillary nerve were observed. SPSS 11.5 software was used to analyze the data.

RESULTSThe length and diameter of cranial middle fossa segment of maxillary nerve were (10.70 ± 1.31) mm and (4.01 ± 0.52) mm respectively, which was supplied by inferior-lateral cavernous sinus artery. The length and diameter of pterygopalatine fossa segment were (16.21 ± 1.80) mm and (3.27 ± 0.62) mm respectively, in which one zygomatic branch, one to three posterior superior alveolar nerves, two ganglion branches and tuberal descending branches; were given off, and the segment was supplied by foramen rotundum artery. The length and diameter of infraorbital segment were (25.73 ± 2.03) mm and (3.30 ± 0.52) mm and it gave off middle superior alveolar nerve (64%) and anterior superior alveolar nerve and was supplied by infraorbital artery. Facial segment gave off superior labial branches, internal and external nasal branches, inferior palpebral branches, buccal branch and zygomatic branch and these branches were supplied by infraorbital artery and superior labial and angular artery originating from facial artery.

CONCLUSIONSUnderstanding of travel and artery supply of maxillary nerve is helpful to regional anaesthesia and surgery for maxillary nerve. Foramen rotundum, sphenopalatine foramen and infraorbital nerve are important marks for endoscopic surgery in pterygopalatine fossa.

Adult ; Cavernous Sinus ; anatomy & histology ; Humans ; Maxillary Artery ; anatomy & histology ; Maxillary Nerve ; anatomy & histology ; blood supply

OBJECTIVETo screen the proteins interacting with inhibitor of differentiation 1(Id1) using yeast two-hybrid analysis in adult human lung cDNA libraries.

METHODSThe coding sequence of Id1 was amplified by PCR and cloned into the bait plasmid. The recombinant bait vector pHybLex/Zeo-Id1 was verified by restriction endonuclease digestion before transformation into the yeast strain EGY48/pSH18-34, which was tested subsequently for reporter genes Leu2 and LacZ activation. The pHybLex/Zeo-Id1 plasmid and the cDNA library plasmid were sequentially transformed into the yeast strains and screened to obtain Leu2(+) and Leu2(+)LacZ(+) clones, with the false positive clones excluded using positive and negative controls, and the plasmid of the true positive clone was sequenced and blasted for homological analysis.

RESULTSSuccessful construction of pHybLex/Zeo-Id1 was confirmed by enzyme digestion. After transformation of pHybLex/Zeo-Id1 into EGY48/pSH18-34, no specific reporter genes Leu2 and LacZ activation was found. The pHybLex/Zeo-Id1 plasmid and the cDNA library plasmid were sequentially transformed into yeast strain, and 198 Leu(+) clones and 19 Leu(+)LacZ(+) double positive clones were obtained. After elimination of the false positive clones, one true positive clone was obtained, whose plasmid analysis by sequencing and blasting indicated high homology (99.5%, 556/559) to AGGF1 (an angiogenic factor with G-patch and FHA domains 1). AGGF1 expression was confirmed in the true positive yeast cells by Western blotting.

CONCLUSIONAGGF1 is confirmed to interact with Id1 by yeast two-hybrid analysis for screening adult human lung cDNA libraries.

Adult ; Angiogenic Proteins ; genetics ; metabolism ; Cell Proliferation ; Endothelial Cells ; cytology ; metabolism ; Gene Library ; Humans ; Inhibitor of Differentiation Protein 1 ; metabolism ; Lung ; cytology ; Plasmids ; genetics ; Protein Binding ; Two-Hybrid System Techniques

OBJECTIVETo construct full-length hepatitis B core particles presenting preS1 aa 21-47 epitope and truncated core particles presenting preS1 aa 37-45 epitope on their surface and compare their antigenicity.

METHODSPreS1 aa21-47 epitope and aa 37-45 epitope were inserted respectively into full-length hepatitis B core (aa 1-183) and truncated HBcAg (aa 1-144), between the 78th (Asp) and 79th (Pro). The genes synthesized after the codon optimization were ligated to the pET43. 1a vector with the same cohesive terminal (NdeI and XhoI) and expressed in the E. coli expression system. The morphology of the proteins of interest were observed by electron microscope and characterized by ELISA and Western Blotting.

RESULTSThe morphology of the virus-like particles were confirmed by electron microscope. H2 were solid particles with a diameter of (31.61 +/- 1.27) nm, while H3 were hollow particles with a diameter of (28.46 +/- 1.16) nm. Statistical analysis showed that H2 is larger than H3 in the diameter (P < 0.01). The antigenicity of the inserted epitopes and carrier protein were identified by ELISA and Western Blotting.

CONCLUSIONChimeric hepatitis B core particles presenting the preS1 neutralizing epitopes on their surface have been expressed, purified and identified, which lays the foundation for its application in vaccine research.

Epitopes ; chemistry ; genetics ; immunology ; Hepatitis B ; immunology ; virology ; Hepatitis B Core Antigens ; chemistry ; genetics ; immunology ; Hepatitis B Surface Antigens ; chemistry ; genetics ; immunology ; Hepatitis B virus ; chemistry ; genetics ; immunology ; Humans ; Neutralization Tests ; Protein Precursors ; chemistry ; genetics ; immunology ; Recombinant Fusion Proteins ; chemistry ; genetics ; immunology

OBJECTIVETo construct a subtractive cDNA library of genes transactivated by PS1TP1 protein with suppression subtractive hybridization (SSH) technique.

METHODSSuppression subtractive hybridization technique and bioinformatics technique were used, the mRNA from HepG2 cells transfected with pcDNA3.1 (-) -PS1TP1 and pcDNA3. 1 (-) empty vector was isolated, respectively; cDNA underwent two times of nested PCR, amplified cDNA fragments were subcloned into pGEM-Teasy vectors to set up the subtractive library.

RESULTSThe subtractive library of genes transactivated by PS1TP1 was constructed successfully. Sequence analysis was performed in 43 clones randomly, and the full length sequences were obtained with bioinformatics method and searched for homologous DNA sequence from GenBank, altogether 12 coding sequences were gotten, which consisted of 10 known and 2 unknown ones.

CONCLUSIONThe obtained sequences may be target genes transactivated by PS1TP1 protein among which some genes coding proteins involved in cell cycle regulation, metabolism, immunity and cell apoptosis. This finding brought some clues for studying the biological functions of PS1T1.

Hepatitis B ; genetics ; Humans ; Nucleic Acid Hybridization ; methods ; Transcriptional Activation ; genetics ; Viral Proteins ; genetics

OBJECTIVETo study the antigenic properties of mutant hepatitis B virus surface antigen, to understand the sensitivity of the commercially available HBsAg assays to the variants and to reduce the undetectability of the variants.

METHODSRecombinant eukaryotic expression plasmids for HBsAg. The recombinant eukaryotic expression plasmids pSS1adr, pSS1adw2, pSS1adw2- 145Arg, pSS1adr-126 Asn and pSS1adr-126Ser were transfected into COS-7 cells. HBsAg in the supernatants of transfected cells was detected by using different commercial ELISA kits.

RESULTSThe absorbance value of pSS1adr-126 Asn and pSS1adr-126Ser plasmids were similar to that of the wild type HBsAg, the absorbance value of pSS1adw2-145Arg plasmids was lower than that of the wild type HBsAg.

CONCLUSIONIt is estimated that the antigenicity of HBsAg mainly depended on the amino acid sequence of "a" antigen determinant and its conformation, so 145 amino acid substitutions led to the change of conformation and the antigenicity of variant HBsAg was lower than that of the wild type.

Animals ; COS Cells ; Cercopithecus aethiops ; Culture Media, Conditioned ; metabolism ; Enzyme-Linked Immunosorbent Assay ; Genetic Vectors ; genetics ; metabolism ; Hepatitis B Surface Antigens ; analysis ; genetics ; immunology ; Mutation ; Transfection ; Viral Envelope Proteins ; genetics

OBJECTIVETo determine the main genotype of hepatitis B virus (HBV) in Xinjiang.

METHODS200 HBsAg positive serum specimens were detected from more than 2000 serum of Xinjiang inhabitants, and HBV S gene was detected by using nPCR amplifying, and compared with the standard S region HBV nucleotide sequences of genotypes A-H retrieved from GenBank, then analyzed and drawn the polygenetic tree by MEGA3 software.

RESULTGene in 127 (63.5%) serum specimens was detected from 200 samples. Among 127 serum specimens, 10 (7.8%) was genotype B, 58 (45.7%) was genotype C, and 59 (46.5%) was genotype D.

CONCLUSIONGenotype B, C and D have been found in Xinjiang.

Adolescent ; Adult ; Aged ; Child ; China ; Female ; Genotype ; Hepatitis B ; ethnology ; virology ; Hepatitis B virus ; genetics ; isolation & purification ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; Rural Health ; Young Adult

OBJECTIVE To determine the characterization, anti-tumor efficacy and pharmacokinetics of bufalin- loaded PEGylated liposomes compared with bufalin entity. METHODS Bufalin- loaded PEGylated liposomes and bufalin- loaded liposomes were prepared reproducibly with homogeneous particle size by the combination of thin film evaporation method and high pressure homogenization method. The particle size and zeta potential of the liposomes were determined by dynamic light scattering technique. The direct imaging of morphology of liposomes was charactered by transmission electron microscope. The content of bufalin in liposomes was analysed by HPLC method. The entrapment efficiency and the particle size was applied to assess the stability profile, after storage at 4℃ on day 0, 7, 15, 30 and 90. The in-vitro release behaviours of bufalin from liposomes were conducted using dialysis bag technique at 37℃. In-vitro cytotoxicity studies were carried out using MTT〔3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide〕assay on several kinds of tumor cell lines including SW620, PC-3, MDA-MB-231, A549, U251, U87 and HepG2. In-vivo pharmacokinetic study of bufalin liposomes was evaluated by HPLC method. RESULTS Their mean particle sizes were 127.6 nm and 155.0 nm, mean zeta potentials were 2.24 mV and - 18.5 mV, entrapment efficiencies were 76.31% and 78.40% , respectively. In- vitro release profile revealed that the release of bufalin in bufalin- loaded PEGylated liposomes was slower than that of bufalin-loaded liposomes. The cytotoxicity of blank liposomes has been found within acceptable range, whereas bufalin-loaded PEGylated liposomes showed enhanced cytotoxicity to U251 cells compared with bufalin entity. In-vivo pharmacokinetics indicated that bufalin-loaded PEGylated liposomes could extend eliminate half-life time of bufalin in plasma in rats. CONCLUSION The results suggested that bufalin-loaded PEGylated liposomes improved the solubility and increased the drug concentration in plasma.