This paper reviews the binding mode beween deoxyribonucle ic acid (DNA) and the different type of fluorescence probe. The application of t hese probes in DNA quantitation analysis and other ways is also reported. The pr ogress of DNA fluorescence probe is addressed.

Astragalus Plant ; chemistry ; Drugs, Chinese Herbal ; pharmacology ; Genes, MHC Class I ; genetics ; HL-60 Cells ; Humans ; K562 Cells ; Killer Cells, Natural ; drug effects ; Leukemia ; genetics ; metabolism ; Polysaccharides ; pharmacology

OBJECTIVE:To clone,prokaryotic express and characterize the TEM-type ?-lactamase produced by Enterobacter cloacae clinical isolate EC002. METHODS: The drug susceptibility of Enterobacter cloacae clinical isolate EC002 was detected by agar double dilution,double disk screening and confirmatory test were employed to detect the ESBLs. The isoelectric point (pI) of enzyme was detected by isoelectric focusing electrophoresis (IEF),the genes were coded by PCR amplification enzyme,and the prokaryotic expression and phenotype of the TEM-type ?-lactamase were detected. RESULTS: Enterobacter cloacae EC002 were resistant to most of the ?-lactamases. Positive results were noted for the phenotype identification and plasmid conjugation test. IEF showed that Enterobacter cloacae EC002 produced two ?-lactamases with pI value at 8.7 and 5.4 respectively,which were confirmed to be CTX-M-22 and a new TEM-subtype ?-lactamase by DNA sequencing,and the phenotype of the expressed enzyme of the cloned strains was non-ESBLs. The TEM-type ?-lactamase was named as TEM-141 by GenBank. CONCLUSION: The TEM-141 produced by Enterobacter cloacae EC002 was a new type of plasmid-mediated broad-spectrum ?-lactamase.

The objective of this study was to explore the effect of astragalus polysaccharide (APS) on sensitivity of leukemic cell line HL-60 to NK cell cytotoxicity and its mechanism. The cytotoxicities of NK cells against HL-60 cells were analyzed by LDH releasing assay at different effect-to-target cell ratios (E:T) before and after treated with APS. The gene expression of MHC class I chain-related (MICA) in HL-60 cells before and after APS treatment was assayed with RT-PCR. Protein expression of MICA in HL-60 cells was assayed by flow cytometry before and after treated by APS. The results showed that after treated with APS 15 mg/ml for 48 h, the cytotoxicities of NK cells against HL-60 cells enhanced at different effect-to-target (P < 0.05), and the gene and protein expressions in MICA of HL-60 cells were up-regulated (P < 0.05). It is concluded that the APS can obviously up-regulate the expression of MICA in HL-60 cells, thus enhance sensitivity of HL-60 cells to cytotoxicity of NK cells.
Astragalus Plant ; Cytotoxicity, Immunologic ; drug effects ; HL-60 Cells ; Histocompatibility Antigens Class I ; metabolism ; Humans ; Killer Cells, Natural ; Polysaccharides ; pharmacology

OBJECTIVETo investigate the absolute bioavailability of caffeic acid in rats and its intestinal absorption properties.

METHODThe absolute bioavailability (Fabs) of caffeic acid was obtained after iv (2 mg x kg(-1)) or ig (10 mg x kg(-1)) administration to rats. The intestinal absorption of caffeic acid was explored by the recirculating vascularly perfused rat intestinal preparation. Caco-2 cell model was applied to measure the permeability of caffeic acid from apical to basolateral said (A-B) and from basolateral to apical said (B-A).

RESULTA two-compartment pharmacokinetic model was best to describe the pharmacokinetics of caffeic acid following iv or ig administration. The Fabs of caffeic acid was 14. 7% , and its intestinal absorption was 12.4%. The values of Papp A-->B and Papp B-->A of caffeic acid were retained stable while its concentration was changed. The efflux ratio values in this study surveyed were above 2.0, and suggesting caffeic acid was active transport.

CONCLUSIONCaffeic acid was shown to have poor permeability across the Caco-2 cells, low intestinal absorption and low oral bioavailability in rats.

Animals ; Biological Availability ; Caco-2 Cells ; Caffeic Acids ; metabolism ; pharmacokinetics ; Humans ; Intestinal Absorption ; Male ; Rats ; Rats, Sprague-Dawley

In order to explore the clinical hypolipidemic features of Daidai flavone extract, the pharmacokinetics features of characteristic active ingredients of Daidai flavone extract in normal and hyperlipemia rats were studied and compared. The study established the quantitative determination method of naringin and neohesperidin in plasma by UPLC-MS. Study compared the pharmacokinetics differences of naringin and noehesperidin in normal and hyperlipemia rats on the basis of establishment of hyperlipemia model. Results indicated that the pharmacokinetics features of characteristic active ingredients of Daidai flavone extract in normal and hyperlipemia rats showed significant differences. The C(max) of naringin and neohesperidin in hyperlipemia rats plasma after oral administration of Daidai flavone extract increased obviously, while t1/2, MRT and AUC0-24 h decreased, compared to normal rats. But t(max) showed no differences to that of normal rats. The results further proved Daidai flavone extract would have better hypolipidemic effect in the hyperlipemia pathological status. And the characteristic active ingredients naringin and noehesperidin were the material base of Daidai flavone extract to express the hypolipidemic effect.
Animals ; Citrus ; chemistry ; Drugs, Chinese Herbal ; isolation & purification ; pharmacokinetics ; Flavones ; chemistry ; Hyperlipidemias ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley

The effect of magnetic Fe3O4 particles on cellulase in the enzymatic hydrolysis of sunflower seed hull was studied in different adding ways and additive amount. In the process of enzymatic hydrolysis of sunflower seed hull, the variations of cellulase activity, reducing sugar concentration and cellulose conversion were evaluated. After the reaction, the analysis of pH and surface tension of hydrolysate were also used to determine the mechanisms of cellulase by the magnetic effect. The results indicated that after adding magnetic Fe3O4, the cellulase activity, reducing sugar concentration and conversion of cellulose had an increased between the 0.5 g/L and 2.0 g/L cases after 48 h. When the additive amount of magnetic Fe3O4 was 2 g/L, the cellulase activity at 60 h was improved significantly by 25.9%. It was found that the concentration of reducing sugar was increased from 6.950 mg/mL to 8.775 mg/mL with magnetic Fe3O4 1.5 g/L. Simultaneously, compared with the blank, which the conversion of cellulose was 47.932%, the maximum celluloseconversion of samples with adding magnetic Fe3O4 was 60.531%. Besides, the stability of cellulase activity adding in times was better than in one time. After the reaction, the final surface tension of hydrolysate with 1.5 g/L magnetic Fe3O4 was the lowest in comparison with the blank. However, no significant differences were observed in the final pH of the hydrolysate.

This study was aimed to investigate the immunological effect of modified dendritic cells (DC) which inducing cytotoxic T cells (CTL) against lymphoma cells. The DC were isolated from the lymph node and peripheral blood of patients with diffuse large B cell lymphoma (DLBCL). DC were transfected with recombinant adenovirus vector carrying human p53 gene (rAd-p53-DC). The expression of p53 gene was detected by flow cytometry. Western-blot was used to detect the expression of P53. ELISA was used to detect IL-12 level in supernatant. The mixed lymphocyte reaction (MLR) was used to detect the proliferative ability of auto-lymphocyte stimulated by DC. The lactate dehydrogenase (LDH) release test was used to determine the cytotoxicity of CTL. The results indicates that the expressions of DC surface molecule (except for CD1a) such as CD83, CD80, CD86 and HLA-DR were significantly higher in experiment group than that in control group and blank control group. The secretion of IL-12 in supernatant was higher in experiment group than that in control group. The autologous T lymphocyte proliferation and cytotoxic activity against the same kind of DLBL-cells increased in experiment group as compared with control group and blank control group (P < 0.05). The ability to stimulate T lymphocyte proliferation increased with the rising of the ratio of DC and T lymphocyte. However, there was statistically significant difference between rAd-p53-DC derived from Lymph node and peripheral blood (P < 0.05). It is concluded that rAd-p53-transfected DC can induce CTL response in vitro against lymphoma cells.
Adenoviridae ; Cell Line, Tumor ; Dendritic Cells ; cytology ; immunology ; Genes, p53 ; Genetic Vectors ; Humans ; Lymphocyte Activation ; Lymphocyte Culture Test, Mixed ; Lymphoma, Large B-Cell, Diffuse ; blood ; immunology ; Transfection

OBJECTIVETo compare the efficacy, side effects and influence of two chemotherapy regimens, paclitaxel liposome combined with platinum and paclitaxel combined with platinum, on the survival rate in patients with cervical carcinoma receiving concurrent chemoradiotherapy.

METHODSOne hundred and sixty two cases with primary cervical carcinoma diagnosed and treated in the Jiangxi Maternal and Children Hospital between January 2008 and November 2009 were enrolled in this randomized controlled trial. Seventy one cases were included in the paclitaxel group and 91 in the paclitaxel liposome group. The chemotherapy doses were as followings: paclitaxel liposome and paclitaxel 135 mg/m(2); cisplatin 80 mg/m(2) or carboplatin AUC 4 - 6, repeated every 21 days for two or three times. Radical radiotherapy was given to both groups at the same time. The efficacy was evaluated by the tumor regression and the patients were followed-up for six months.

RESULTSThe overall response rates of paclitaxel group and paclitaxel liposome group were 90.1% and 89.0%, respectively (P > 0.05). The 1-year cumulative survival rate was 91.4% for the paclitaxel group and 89.2% for the paclitaxel liposom group (P > 0.05). The incidence rate of adverse effects such as rash, gastrointestinal toxicity, bone marrow suppression and muscle/joint pain in the paclitaxel liposome group was significantly lower than that in the paclitaxel group (P < 0.05), while there was no significant difference regarding the hair loss, liver damage, and peripheral neuritis (P > 0.05).

CONCLUSIONSPaclitaxel liposome plus platinum is a safe and effective therapeutic regimen for stage IIa-IV cervical carcinoma. However, the long-term efficacy of this regimen should be further observed.

Adenocarcinoma ; pathology ; therapy ; Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; adverse effects ; therapeutic use ; Brachytherapy ; Carboplatin ; administration & dosage ; adverse effects ; Carcinoma, Squamous Cell ; pathology ; therapy ; Chemoradiotherapy ; Cisplatin ; administration & dosage ; adverse effects ; Cobalt Radioisotopes ; therapeutic use ; Exanthema ; chemically induced ; Female ; Follow-Up Studies ; Gastrointestinal Diseases ; chemically induced ; Humans ; Iridium Radioisotopes ; therapeutic use ; Liposomes ; administration & dosage ; adverse effects ; Middle Aged ; Neoplasm Staging ; Paclitaxel ; administration & dosage ; adverse effects ; Remission Induction ; Survival Rate ; Uterine Cervical Neoplasms ; pathology ; therapy

AIMTo investigate the role of TNF-alpha in vascular endothelial cells injury mediated by freezing/thaw ing PMN.

METHODSFreezing/thawing cell model was founded using rat PMN isolated by dextran sedimentation technique and VEC cultured in vitro. The injury level of VEC was indicated by measuring activity of LDH in medium. The number of frozen/thawed PMN adhering to VEC was counted with Phagocytizing reactive dyes the degree of frozen/thawed PMN and VEC adhesion. Expression of LFA-1 on the surface of frozen/thawed PMN was analyzed with flow cytometry.

RESULTSTNF-alpha could obviously upregulate expression of LFA-1 on surfaced of frozen/thawed PMN. Upregulation of LFA-1 expression promoted adhesion of frozen/thawed PMN and normal VEC,and aggravated VEC injury. Monoclonal antibody against LFA-1 could partly block adhesion of frozen/thawed PMN and normal VEC,and attenuate VEC injury.

CONCLUSIONTNF-alpha can promote expression of LFA-1 on surface of frozen/thawed PMN adhering of frozen/thawed PMN to normal VEC and VEC injury increase, monoclonal antibody against LFA-1 could partly block PMN-VEC adhesion and attenuate VEC injury.

Animals ; Cell Adhesion ; Cells, Cultured ; Endothelial Cells ; drug effects ; Endothelium, Vascular ; cytology ; Freezing ; Lymphocyte Function-Associated Antigen-1 ; metabolism ; Neutrophils ; cytology ; metabolism ; Rats ; Tumor Necrosis Factor-alpha ; pharmacology