Objective:To construct a fusion DNA vaccine pEGFP-C3-B7.2-hTERT and to observe the capability of pEGFP-C3-B7.2-hTERT DNA vaccine to induce specific anti-tumor immune responses and to inhibit growth of the hepatoma transplanted from H22 cells. Methods: The B7.2 and hTERT cDNA (amplified by RT-PCR), together with pEGFP-C3 as the vector were used to construct fusion gene plasmid pEGFP-C3-B7.2-hTERT, which was then used to vaccinated C57BL/6 mice for 3 times at a 7 d interval. Animals vaccinated with pEGFP-C3-B7.2, pEGFP-C3-hTERT, pEGFP-C3 and PBS were taken as controls. Splenocytes CTLs of immunized mice, the levels of IL-2, IFN-? in the culture supernatant, the levels of antibodies against hTERT, and the changes of the T lymphocyte subsets in the peripheral blood were all examined. The mice harboring hepatocarcinoma H22 cells were challenged with pEGFP-C3-B7.2-hTERT and the tumor forming time and the survival periods of mice were observed. Results: Agarose gel electrophoresis, nuclease digestion and sequencing confirmed the successful construction of pEGFP-C3-B7.2-hTERT. The CTLs activity of splenocytes from the mice immunized with pEGFP-C3-B7.2-hTERT fusion gene was significantly higher than those of the other groups (P

Adult ; Female ; Hemofiltration ; Hemolysis ; drug effects ; Humans ; Male ; Methylprednisolone ; administration & dosage ; therapeutic use ; Middle Aged ; Plasma Exchange ; Treatment Outcome ; Wasp Venoms ; poisoning ; Young Adult

Objective:To construct pEGFP-C3-B7.2-MAGE-1 eukaryotic expression plasmids and to study the immunological effects of esophageal cancer cell vaccine modified by human B7.2/MAGE-1 gene.Methods:The B7.2 and MAGE-1 cDNA,which was amplified by RT-PCR, and the eukaryotic expression plasmid pEGFP-C3-B7.2-MAGE-1 was constructed. Human esophageal cancer cell EC9706 was transfected with the vector of pEGFP-C3-B7.2-MAGE-1 using the technique of lipofectamine transfection.The dendritic cells(DCs)from peripheral blood mononuclear cells(PBMC)were loaded with the tumor antigen,and co-cultured with congeneric T cells derived from PBMCs for 3 days to obtain the tumor specific cytotoxicity T lymphocytes(CTL). Methy1 thiazoly1 tetrazolium (MTT) assay was used to detect inhibition effect of CTL on transfected and untransfected EC9706 cells.Results:The CTL had stronger inhibition response against the cancer cells transfected with pEGFP-C3-B7.2-MAGE-1 than to the cancer cells transfected with pEGFP-C3 and the untransfected cancer cells(P

Biopsy ; Eosine Yellowish-(YS) ; Gastric Mucosa ; microbiology ; pathology ; Helicobacter Infections ; diagnosis ; pathology ; Helicobacter pylori ; isolation & purification ; Hematoxylin ; Humans ; Silver Staining ; Staining and Labeling ; methods

Aim To study the mechanism of the anti-aging effect of total extract of astragalus(TEA) in mice. Methods MDA content, the activities of Mn-SOD and GSHpx and the reduced to oxidized glutathione (GSH/GSSG) ratio in mitochondria of D-galactose(D-gal) treated mice and 17-month-old mice were measured. Results Treatment with TEA(40 mg?kg-1?d-1 ig) for 10 wk would lower the content of MDA and restore activities of Mn-SOD,GSHpx and GSH/GSSG ratio in mitochondria of D-gal treated mice.Treatment with TEA (40 mg?kg-1 ?d-1 ig) for 3 mon had the same effect on 17-month-old mice.Conclusion TEA has anti-aging effect on D-gal treated mice and 17-month-old mice significantly, probably being related to its anti-oxidative effect.

Administration, Oral ; Adult ; Atrial Flutter ; drug therapy ; physiopathology ; Digoxin ; therapeutic use ; Echocardiography ; Female ; Fetal Diseases ; drug therapy ; physiopathology ; Humans ; Hydrops Fetalis ; drug therapy ; physiopathology ; Infant, Newborn ; Male ; Pregnancy

Objective To explore the association of HbA1C with microvascular complications,and to evaluate the diagnostic value of HbA1C in diabetes mellitus in high-risk populations of Guangzhou.Methods HbA1C,blood glucose,fundus photography,and microalbuminuria were detected in 208 permanent residents with high-risk factors of diabetes.The receiver operating characteristiC(ROC)curves were used to estimate the area of HbA1C,fasting plasma glucose(FPG),postprandial 2 h plasma glucose(2hPG)under the curve for discriminating microvascular complications.Results There were 14.9% adults suffering from diabetic retinopathy and 12% microalbuminuria in high risk populations of diabetes.The optimal cutoff points of HbA1C,FPG,and 2hPG in detecting retinopathy were 5.8%,7.0 mmol/L,and 10.9 mmol/L respectively.The thresholds for increasing prevalence of microalbuminuria were5.8% for HbA1C,6.4 mmol/L for FPG,and 10.7 mmol/L for 2hPG.Conclusions The prevalence of diabetic microvascular complications increases dramatically at the concentration of HbA1C 5.8%.As a diagnostic value for microvascular complications,there is no significant difference between HbA1C and 2hPG.

OBJECTIVETo study the chemical constituents of ethanolic extract from herbs of Dicliptera chinensis.

METHODThe compounds were separated by silica gel column chromatography, preparation TLC and reverse phase HPLC, their structures were identified by the spectroscopic methods of UV, NMR and MS.

RESULTSeven compounds were isolated from ethyl acetate extract. Their structure were identified as octasulphur (1), secoisolariciresinol dimethyl ether diacetate (2), 5-methoxy-4, 4'-di-O-methyl secolariciresinol (3), chinensinaphthol methyl ester (4), loliolide (5), beta-sitosterol 3-O-beta-D-glucopyranoside (6) and stigmasterol 3-O-beta-D-glucopyranoside (7).

CONCLUSIONAll the compounds except 6 were obtained from the plants of Dicliptera for the first time.

Acanthaceae ; chemistry ; Benzofurans ; chemistry ; isolation & purification ; Glucosides ; chemistry ; isolation & purification ; Plants, Medicinal ; chemistry ; Sitosterols ; chemistry ; isolation & purification

AIMTo investigate the influence and the mechanism of SK on the contractility of cultured cardiomyocytes of rats.

METHODSThe primary cultured single myocardial cell was treated with SK and the contraction frequency and size of cardiomyocyte were determined by a computer image analysis system. At the same time the effects of propranolol (a beta receptor antagonist), phentolamine (a alpha receptor antagonist), DSP (a tachykinin receptor antagonist) on the action of SK were investigated.

RESULTSSK increased contractive extend of the cardiomyocyte, in which a dose-response relationship of SK at 1.78 x 10(-8) - 1.78 x 10(-5) mol/L exists. But the frequency of contraction did not change, pretreatment with propranolol, phentolamine had no action on the effect of SK, but DSP markedly attenuated the effects of SK.

CONCLUSIONSK may directly enhance the contractility of single cardiomyocyte, which may be related with the tachykinin receptor.

Animals ; Animals, Newborn ; Benzylamines ; pharmacology ; Cells, Cultured ; Dose-Response Relationship, Drug ; Myocardial Contraction ; drug effects ; Myocytes, Cardiac ; drug effects ; physiology ; Neurokinin A ; pharmacology ; Phentolamine ; pharmacology ; Propranolol ; pharmacology ; Rats ; Rats, Wistar ; Receptors, Tachykinin ; antagonists & inhibitors

OBJECTIVETo establish a HPLC method for determination of protopine and isocorydine in root of Dactylicapnos scandens.

METHODThe separation was performed in a PHENOMENEX-C18 column with a mobile phase of methanol-0.2% phosphoric acid (adjusted to pH 7.0 with triethylamine)(50:50), The detection wavelength was at 254 nm and the flow rate was 0.8 mL x min(-1).

RESULTThe average recovery of Protopine and Isocorydine was 97.9%, 98.6% respectively, and RSD 1.3%, 1.4%.

CONCLUSIONThis method is accurate, simple and reliable. It can be used for quality control of D. scandens.

Aporphines ; analysis ; Benzophenanthridines ; Berberine Alkaloids ; analysis ; Chromatography, High Pressure Liquid ; Papaveraceae ; chemistry ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Quality Control