Animals ; Calmodulin ; metabolism ; Epimedium ; Flavonoids ; pharmacology ; Gene Expression ; Hypothalamo-Hypophyseal System ; drug effects ; metabolism ; Male ; Pituitary-Adrenal System ; drug effects ; metabolism ; Rats ; Rats, Sprague-Dawley

Study the immunological adjuvant function of biodegradable microspheres for DNA immunization. Empty poly (D, L-lactide-co-glycolic acid) microspheres were prepared using the water- inoil-in-water (w-o-w) technique; A plasmid DNA pRc-CMV encoding hepatitis B virus S antigen was constructed; The mixture of the microspheres and the plasmid DNA was prepared by incubation method. The mixture was administered to Balb/c mice by intramuscular injection. Result: The high antibody titer(1:1600) of intramuscular injection of the mixture of microspheres and the plasmid DNA was obtained, similar to that of intramuscular injection of the mixture of AL(OH)3 and hepatitis B virus S antigen; while intramuscular injection the plasmid DNA elicited no serum antibody respones. Conclusion: biodegradable microspheres may be used as an good adjuvant for DNA immunization.

Objective To report a peliosis hepatic in child and review literature and discuss.Methods Case history was inquired.Physical,labtoratory,imagement and histopathology of liver biopsy(HE staining) were examed.Results A 4-year old girl appeared dermatitis with erythema and herpes at local skin where was bit by insect before onset.The girl appeared fever,cough,then abdominal pain,hepatomegaly,pleural effusion and ascites.Lab examination revealed slight elevation of aspartate transaminase,?-glutamyltranspeptidase and alkaline phosphatase.The liver B-mode ultrasonography and CT scan revealed hepatomegaly with density heterogeneity of the parenchyma.The liver biopsy revealed many small capsule filled with blood cells.Conclusions Clinical characteristics of the disease are fever,upper abdomen pain,janundice,ascites and hepatomegaly.The diagnosis shall be combined with the pathologic biopsy of liver.

BACKGROUNDThe synthesis of virus-like particles (VLPs) provides an important tool to determine the structural requirements for viral particle assembly and virus-host interactions. Our purpose was to express simultaneously all three structural proteins of hepatitis C virus (HCV) in insect cells to investigate the proteins assembly into VLPs and the immunogenicity of these particles.

METHODSHCV gene sequences encoding the structural proteins C, E1, and E2 were amplified with PCR, and recombinant baculoviruses were constructed using recombinant DNA techniques. The expression of HCV structural proteins in insect cells was analyzed by immunofluorescence and SDS-PAGE. The interaction of expressed structural proteins was investigated by immunoprecipitation and immunoblotting. The VLPs in the insect cells were visualized by electron microscopy (EM). VLPs were then purified by sucrose gradient centrifugation and used to immunize BALB/c mice. Antibodies against HCV were tested for in mouse serum samples by an ELISA assay.

RESULTSThe recombinant baculoviruses reBV/C and reBV/E1-E2 were constructed successfully. Insect cells co-infected with reBV/C and reBV/E1-E2 expressed HCV C, E1, and E2 proteins with the expected molecular weights of 20kD, 35kD, and 66kD, respectively. The results of immunoprecipitation and immunoblotting assays revealed the coimmunoprecipitation of C, E1, and E2 proteins, indicating association of the three structural proteins. Electron microscopy of insect cells co-infected with reBV/C and reBV/E1-E2 demonstrated spherical particles (40 to 60 nm in diameter) similar to the HCV virions from serum samples or hepatic tissue samples of HCV infected humans. The VLPs were partially purified. Antibodies to HCV were detectable in the serum of mice immunized with VLPs.

CONCLUSIONHCV structural proteins simultaneously expressed in insect cells can interact with each other and assemble into HCV-like particles, which are shown to be immunogenic in mice.

Animals ; Fluorescent Antibody Technique ; Hepacivirus ; immunology ; physiology ; Immunization ; Immunoprecipitation ; Mice ; Mice, Inbred BALB C ; Microscopy, Electron ; Spodoptera ; Viral Structural Proteins ; immunology ; physiology ; Virion ; physiology ; Virus Assembly

OBJECTIVETo explore the significance of mitochondrial D-loop alterations in hyperplastic pancreatic ductal cells in vicinity of pancreatic cancer coexisting with chronic pancreatitis.

METHODSMalignant lesions and foci of pancreatic ductal intraepithelial neoplasia of the pancreas and paired normal gastric mucosal epithelial cells from the same patients, respectively, were assessed by polymerase chain reaction. Somatic point mutations and sequence variants of D-loop were searched by direct sequencing of the mitochondrial genome. D-loops were sequenced by BLAST to identify their mutations.

RESULTSEleven of 12 pancreatic cancers displayed at least one D-loop variants and one tumor presented heteroplasmy. There was an apparent increase in incidence of D-loop mutational rate from PanIN1 (33.3%) to PanIN3 (75%, P < 0.01).

CONCLUSIONMitochondrial D-loop alterations in the pancreas occur in the earliest premalignant lesions and exhibite an increasing occurence that parallels histological severity. These alterations may serve as a valuable marker to follow the histopathological progression of the lesions. Large number of further studies are required to clarify clinical implications of the mitochondrial DNA alterations.

Adenoma ; complications ; genetics ; Adult ; Aged ; Base Sequence ; DNA, Mitochondrial ; genetics ; Epithelial Cells ; metabolism ; pathology ; Female ; Humans ; Male ; Middle Aged ; Mutation ; Pancreatic Ducts ; metabolism ; pathology ; Pancreatic Neoplasms ; complications ; genetics ; Pancreatitis, Chronic ; complications ; genetics ; Precancerous Conditions ; complications ; genetics ; Sequence Analysis, DNA

Glycoproteins ; genetics ; metabolism ; Herpesviridae ; genetics ; metabolism ; Viral Proteins ; genetics ; metabolism

It is a challenging and important project to prolong the in vivo half life of protein and peptide drugs by physicochemical methods without new molecular entities generation. Protein crystallization provides a new strategy for improving the stability and in vivo delivery of these drugs. We show here that recombinant human interferon-alpha (rhIFN) can form spherical crystals. The physical and chemical features of the crystals were characterized, and drug dissolution was determined in vitro. The pharmacokinetics of crystallized interferon after sc injection in rabbit at 1.5 x 10(7) U x kg(-1) was compared to that of soluble form. The crystals were characterized as mono-dispersed spheres, with yield of > 80%, mean diameter size of about 16 microm and crystallinity of 23.2%. The in vitro dissolution behavior of crystallized rhIFN was featured as low initial burst release (21% within the first 2 h) and prolonged cumulative dissolution time up to 72 h without biological potency lost. After sc administration of soluble and crystallized interferon in rabbits, the peak time (T(max)) and half life (t1/2) were prolonged from (1.80 +/- 0.45) h and (1.35 +/- 0.35) h to (13.20 +/- 2.68) h and (10.68 +/- 1.97) h, respectively. The corresponding peak concentration decreased from (1 411.10 +/- 575.28) U x mL(-1) to (721.37 +/- 206.55) U x mL(-1). PK/PD analysis indicated that (96.87 +/- 20.30) % of relative bioavailability was obtained. The research results of this work will provide important academic value and application prospect for improving clinical therapeutic effect and development of biomacromolecules delivery system for protein and peptide drugs.
Animals ; Antiviral Agents ; administration & dosage ; chemistry ; pharmacokinetics ; Biological Availability ; Crystallization ; Delayed-Action Preparations ; Drug Delivery Systems ; Half-Life ; Humans ; Injections, Subcutaneous ; Interferon-alpha ; administration & dosage ; chemistry ; pharmacokinetics ; Male ; Rabbits ; Recombinant Proteins ; administration & dosage ; chemistry ; pharmacokinetics ; Solubility ; Surface Properties

BACKGROUNDNon-valvular atrial fibrillation is associated with an increased risk of ischemic stroke; however, the appropriate intensity of anticoagulation therapy for Chinese patients has not been determined. The purpose of this study was to compare the safety and the efficacy of standard-intensity warfarin therapy, low-intensity warfarin therapy, and aspirin therapy for the prevention of ischemic events in Chinese patients with non-valvular atrial fibrillation (NVAF).

METHODSA total of 786 patients from 75 Chinese hospitals were enrolled in this study and randomized into three therapy groups: standard-intensity warfarin (international normalized ratio (INR) 2.1 to 2.5) group, low-intensity warfarin (INR 1.6 to 2.0) group and aspirin (200 mg per day) group. All patients were evaluated by physicians at 1, 3, 6, 9, 12, 15, 18, 21 and 24 months after randomization to obtain a patient questionnaire, physical examination and related laboratory tests.

RESULTSThe annual event rates of ischemic stroke, transient ischemic attack (TIA) or systemic thromboembolism were 2.6%, 3.1% and 6.9% in the standard-intensity warfarin, low-intensity warfarin and aspirin groups, respectively (P = 0.027). Thromboembolic event rates in both warfarin groups were significantly lower than that in the aspirin group (P = 0.018, P = 0.044), and there was no significant difference between the two warfarin groups. Severe hemorrhagic events occurred in 15 patients, 7 (2.6%) in the standard-intensity warfarin group, 7 (2.4%) in the low-intensity warfarin group and 1 (0.4%) in the aspirin group. The severe hemorrhagic event rates in the warfarin groups were higher than that in the aspirin group, but the difference did not reach statistical significance (P = 0.101). The mild hemorrhagic and total hemorrhagic event rates in the warfarin groups (whether in the standard-intensity warfarin group or low-intensity warfarin group) were much higher than that in the aspirin group with the annual event rates of total hemorrhages of 10.2%, 7.6% and 2.2%, respectively, in the 3 groups (P = 0.001). Furthermore, there was no significant difference in all cause mortality among the three study groups.

CONCLUSIONIn Chinese patients with NVAF, the warfarin therapy (INR 1.6 - 2.5) for the prevention of thromboembolic events was superior to aspirin.

Aged ; Aged, 80 and over ; Anticoagulants ; administration & dosage ; therapeutic use ; Aspirin ; administration & dosage ; therapeutic use ; Atrial Fibrillation ; drug therapy ; Female ; Humans ; Male ; Middle Aged ; Warfarin ; administration & dosage ; therapeutic use

OBJECTIVETo investigate the changes of gene expression in rat neurons induced by 1.8 GHz radiofrequency electromagnetic fields (RF EMF) and to screen for the RF EMF-responsive genes.

METHODSNewly-born SD rats in 24 hours were sacrificed to obtain cortex and hippocampus neurons. The cells were divided randomly into two groups: the experiment group (the irradiation group) and the control group (the false irradiation group). In the irradiation group, after twelve days' culture, neurons were exposed to 1.8 GHz RF EMF modulated by 217 Hz at a specific absorption rate (SAR) of 2 W/kg for 24 hours (5 minutes on/10 minutes off) while in the false control group, the neurons were put in the same waveguide as in the irradiation group, but were not exposed to any irradiation. The total RNA was isolated and purified immediately after exposure. The affymetrix rat neurobiology U34 assay was used for detecting the changes in gene expression profile according to the manufacturer's instruction. RF EMF-responsive candidate gene was confirmed by using ribonuclease protection assay (RPA).

RESULTSAmong 1200 candidate genes, the expression levels of 34 genes were up or down regulated. Microtubule associated protein 2 (Map2) gene was selected as the candidate and subjected to further analysis. RPA data clearly revealed that Map2 was statistically significantly up-regulated after neurons were exposed to the RF EMF (P < 0.05).

CONCLUSIONThe modulation of gene expression and function of Map2 as a neuron specific cytoskeleton protein is crucial to maintain the normal framework and function of neurons. The finding that 1.8 GHz RF EMF exposure increases the expression of Map2 might indicate some unknown effects of RF EMF on neurons.

Animals ; Animals, Newborn ; Cell Phone ; Cells, Cultured ; Dose-Response Relationship, Radiation ; Down-Regulation ; Electromagnetic Fields ; Female ; Gene Expression ; radiation effects ; Male ; Microtubule-Associated Proteins ; biosynthesis ; genetics ; Neurons ; metabolism ; radiation effects ; Radio Waves ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Up-Regulation

OBJECTIVETo evaluate the blood supply of low density viable area of primary heptocellular carcinoma after transcatheter hepatic artery chemoembolization using lipiodol (LP-TACE), by helical dual-phase CT scanning and three dimensional CT (3DCT).

METHODSThirty-four patients with primary heptocellular carcinoma after LP-TACE were examined by hepatic helical dual-phase CT. 3DCT model of the maximum intensity projection (MIP), surface shaded display (SSD) reconstruction of the hepatic artery and portal vein were simultaneously done in 5 cases.

RESULTSViable tumor areas of 34 cases of primary heptocellular carcinoma after LP-TACE were divided into four types: peripheral, lateral, central and diffused types. Enhanced tumor vessel or tissue in viable tumor area was found during hepatic dual-phase in 17 cases, during hepatic artery-phase only in 8 and hepatic portal vein-phase only in 3. The viable tumor areas were found to have blood supply from the hepatic vein in 2 cases. The viable tumor area unenhanced during hepatic dual-phase was found in 6 cases. In 5 cases, the relation between the viable tumor area and branches of hepatic artery and portal vein was showed by MIP and SSD of hepatic artery and portal vein.

CONCLUSIONHepatic helical dual-phase CT scan with 3DCT is effective in evaluating the blood supply of viable tumor areas and the therapeutic effect of primary heptocellular carcinoma after LP-TACE.

Adult ; Aged ; Carcinoma, Hepatocellular ; Catheterization ; Chemoembolization, Therapeutic ; Contrast Media ; Female ; Hepatic Artery ; Humans ; Iodized Oil ; Liver Neoplasms ; diagnostic imaging ; therapy ; Male ; Middle Aged ; Tomography, Spiral Computed