OBJECTIVETo investigate the effects of different concentrations of lipopolysaccharide (LPS), tumor necrosis factor α (TNFα), interleukin-6 (IL-6), dexamethasone (Dex), and insulin on the mRNA and protein expressions of GPR54 in the MCF7 cell line in vitro.

METHODSMCF7 breasr cancer cells were cultured and treated with different concentrations of LPS (10 and 20 µg/ml), TNFα (20 and 100 ng/ml), IL-6 (10 and 20 ng/ml), Dex (10(-6) and 10(-7) mol/L), and insulin (0.01 and 0.1 IU/L). Those treated with culture fluid only served as controls. The mRNA and protein expressions of GPR54 were measured by real-time PCR and Western blot, respectively, after 6, 24, 48, and 72 hours of treatment.

RESULTSCompared with the blank con- trol, LPS (10 and 20 µg/ml), TNFα (20 and 100 ng/ml), IL-6 (10 and 20 ng/ml), Dex (10(-6) and 10(-7) mol/L), and insulin (0.01 and 0.1 IU/L) significantly increased the expressions of GPR54 mRNA (P < 0.05) and protein (P < 0.05).

CONCLUSIONLPS, TNFα, IL-6, Dex, and insulin evidently increase the expression of GPR54 in the MCF7 cell line, indicating their influence on the function of gonads by regulating the GPR54 level.

Blotting, Western ; Dexamethasone ; administration & dosage ; pharmacology ; Glucocorticoids ; administration & dosage ; pharmacology ; Gonads ; drug effects ; metabolism ; Humans ; Hypoglycemic Agents ; administration & dosage ; pharmacology ; Insulin ; administration & dosage ; pharmacology ; Interleukin-6 ; administration & dosage ; pharmacology ; Lipopolysaccharides ; administration & dosage ; pharmacology ; MCF-7 Cells ; RNA, Messenger ; metabolism ; Real-Time Polymerase Chain Reaction ; Receptors, G-Protein-Coupled ; drug effects ; genetics ; metabolism ; Receptors, Kisspeptin-1 ; Time Factors ; Tumor Necrosis Factor-alpha ; administration & dosage ; pharmacology

Objective To investigate apoptosis of HepG2 ceils after transfecfion with LIGHT gene and interferon-γ. Methods LIGHT gene and interferon-γ were transfected into HepG2 cells by liposome mediated method. The HepG2 cells were divided into group A (transfected with LIGHT gene or interferon-γ), group B (transfeeted with LIGHT gene and interferon-γ) and group C (non-transfection group). The apoptosis rate of the HepG2 cells and the expression of Bcl-2 and Caspase-8 were detected 12, 24, 48 hours after transfeetion. Results (1) The apoptosis rates of HepG2 cells at hour 12, 24 and 48 after transfeetion were 18.8% ± 3.5%, 25.7%± 2.8% and 36.4% ±3.6% in group A, 23.8% ±2.4%, 31.1% ±2.1% and42.5% ±4.5% in group B, and 8.7% ± 2.1%, 9.3% ± 1.6% and 10.9% ± 1.2% in group C. There was a significant difference in apoptosis rate among the 3 groups (F = 15.69, 53.33, 48.28, P < 0.01). (2) The expression of Bcl-2 in HepG2 cells at hour 12, 24 and 48 after transfection was 16.4% ± 5.0%, 13.4% ± 3.5% and 8.6% ± 2.3% in group A, 14.7%±3.8%, 9.1% ±2.0% and 4.6% ±2.0% in group B, and 25.3% ±6. 3%, 19.8% ±4.4% and 10.1% ±3.8% in group C. There was a significant difference in the expression of Bcl-2 among the 3 groups (F = 6.19, 12.29, 5.81, P <0.05). (3) The expression of Caspase-8 at hour 12, 24 and48 after transfection were 19.3% ±2.4%, 27.2% ±1.9% and 33.7% ±3.0% in group A, 22.7% ±2.2%, 30.9% ±3.1% and 38.2% ±3.2% in group B, and 1.2% ±0.8%, 1.8% ±0.6% and 3.2% ±1.5% in group C. There was a significant difference in the expression of Caspase-8 among the 3 groups (F =71.54, 112. 78, I01.61, P < 0.01). Condusions LIGHT gene can signiticanfly promote cell apoptosis through regulating the expression of Bcl-2 and Caspase-8. Interferon-γ enhanced the effect of LIGHT gene on the apoptosis of HepG2 cells.

Objective To observe the clinical efficacy of deproteinized calf blood extractive injection combined with hyperbaric oxygen in patients with severe traumatic brain injury.Methods 74 patients with severe traumatic brain injury from January 2013 to March 2015 in Wenzhou hospital of Chinese traditional medicine were randomly divided into observation group and control group, 37 cases in each group.The control group received hyperbaric oxygen on the basis of conventional therapy, the observation group received deproteinized calf blood extractive injection on the basis of control group.The Glasgow coma scale ( GCS) , Barthel, hs-CRP, TNF-α, IL-6 and prognosis were compared between two groups.Results The GCS score and Barthel index scores post-treatment in observation group were (13.67 ±1.73),(65.73 ±4.02) points, which were higher than (9.66 ±1.24), (50.69 ± 3.76) points in control group, and the difference was significant (P<0.05).The serum hs-CRP, TNF-α, IL-6 post-treatment in observation group were (4.55 ±0.76)mg/L,(1.21 ±0.05)μg/L,(0.21 ±0.01)μg/L, which were better than those of control group (6.43 ±1.01)mg/L,(1.36 ±0.06)μg/L,(0.28 ±0.02)μg/L (P<0.05).The rate of favorable prognosis in observation group was 48.65%, which was higher than that of control group (P<0.05).Conclusion The deproteinized calf blood extractive injection combined with hyperbaric oxygen has the exact efficacy, which was better than hyperbaric oxygen alone in the treatment of severe traumatic brain injury.

Using Trichoderma as an indicative fungus, three antifungal proteins in Triticale Zhongsi 237 seed were purified and characterized. These protein components were considered to be a new Class II chitinase and two kinds of beta-1, 3-glucanases. Chitinase molecular mass was 30.5 kD and enzyme activity was maximal at pH 6.0 and 37 degrees C. Two beta-glucanases molecular masses were 51 kD and 23 kD. N-terminal amino acid sequences of Triticale chitinase share high homology with barley chitinase. In some conditions, the chitinase and beta-glucanases all had strong antifungal activity and were able to inhibit Trichoderma growth synergistically. Moreover, the chitinase and beta-1, 3-glucanases were able to inhibit powdery mildew growth on detached susceptible wheat leaves.
Antifungal Agents ; isolation & purification ; pharmacology ; Chitinases ; isolation & purification ; pharmacology ; Glucan 1,3-beta-Glucosidase ; Molecular Weight ; Plant Proteins ; isolation & purification ; pharmacology ; Seeds ; chemistry ; Trichoderma ; drug effects ; Triticum ; chemistry ; microbiology ; beta-Glucosidase ; isolation & purification ; pharmacology

To investigate the modulation on the P-glycoprotein in the jejunum by combined use of Glycyrrhiza inflata and Kansui with ussing chamber and rt-pcr, Rhodamine 123 (R123), a P-gp substrate and fluorescein sodium (CF), a model drug of non-P-gp substrate transported by a passive diffusion were taken as investigational drugs. Because these two drugs can be easily assayed and widely used in various research fields. The permeability of R123 or CF via Wistar rat jejunum membranes was evaluated by in vitro ussing chamber after oral administration of four different decoctions of Glycyrrhiza inflata and Kansui for 1 week. And the concentration of R123 or CF was determined by the fluorospectrophotometry in the receiving solution. Meanwhile the expression of mdr1a in P-glycoprotein was detected by real-time fluorescent quantitative PCR. After oral administration of combined decoction of the single drug, the absorptive directed permeability of R123 increased significantly (P < 0.01). On the other hand, Kansui and combine decoction of the two drugs also decrease the permeability of secretory directed transport (P < 0.05). No action of Glycyrrhiza inflata was found on the secretory transport of R123 [Papp = (2.56 +/- 0.38) x 10(-5), cm x s(-1)] across the jejunum tissues, while Papp of control group was found [Papp = (2.35 +/- 0.27) x 10(-5), cm x s(-1)]. After oral administration of Kansui decoction for 1 week and 2 weeks, the levels of mdr1a expression in Wistar rats were lower than that of the control group, but there were no significant difference in the results. Meanwhile, Glycyrrhiza inflata had no effect on transport of CF across the jejunum tissues, though the other three groups could decrease the permeability of CF, as compared with control group. Kansui may slightly inhibit P-glycoprotein function in the intestinal membrane. For another, some compositions in Kansui inhibit P-glycoprotein function, and some others strengthen the tight junction between cells in the intestinal membrane to decrease permeability of CF. As the inhibitory action to P-glycoprotein was enhanced by combination of Glycyrrhiza inflata and Kansui, based on the results, it may be one of the mechanisms of creating toxicity once co-administration of Glycyrrhiza inflata and Kansui.
ATP Binding Cassette Transporter, Sub-Family B ; genetics ; metabolism ; ATP-Binding Cassette, Sub-Family B, Member 1 ; antagonists & inhibitors ; pharmacokinetics ; Administration, Oral ; Animals ; Biological Transport ; drug effects ; Cell Membrane Permeability ; drug effects ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacology ; Euphorbia ; chemistry ; Fluorescein ; pharmacokinetics ; Glycyrrhiza ; chemistry ; Intestinal Absorption ; Intestinal Mucosa ; metabolism ; Jejunum ; metabolism ; Male ; Plants, Medicinal ; chemistry ; RNA, Messenger ; metabolism ; Random Allocation ; Rats ; Rats, Wistar ; Rhodamine 123 ; pharmacokinetics

OBJECTIVETo identify possible reasons of rotational mismatch between tibial component and femoral component in total knee arthroplasty and choose a right reference axis for placing the components in the operation and to decrease the complications.

METHODSForty normal Chinese knees were studied. There were 20 men and 20 women, and average age was 34 years (range, 18 - 42 years). The images of cross sections of the distal femur were obtained by spiral CT scanning (0.5 mm thickness). Scan direction was aligned to be in the plane perpendicular to the mechanical axis of the tibia. On the images of the distal femur and the proximal tibia, three baselines for the anteroposterior axis of the femoral component were drawn based on the clinical epicondylar axis and the surgical epicondylar axis and 3 degrees lateral rotated to the posterior condylar surfaces of the femur separately, and a baseline for the anteroposterior axis of the tibial component was drawn based on the medial 1/3 of the tibial tuberosity. The rotational mismatch angles were measured between each component by using the Autocad software.

RESULTSThe mean rotational mismatch angle between tibial component and femoral component is 2.94 degrees for the clinical epicondylar axis, 6.50 degrees for the surgical epicondylar axis and 6.83 degrees for 3 degrees lateral rotation of the femoral component referenced to the posterior condylar axis separately.

CONCLUSIONSLandmarks of each bone were the intrinsic cause of the rotational mismatch in total knee arthroplasty. The clinical epicondylar axis can be chosen for the ideal reference to rotational alignment of the femoral component because of its minimal rotational mismatch between each component.

Adolescent ; Adult ; Arthroplasty, Replacement, Knee ; methods ; Female ; Humans ; Knee Joint ; diagnostic imaging ; physiopathology ; surgery ; Male ; Postoperative Complications ; prevention & control ; Range of Motion, Articular ; Tomography, Spiral Computed

OBJECTIVETo explore the effect of donor-derived NK cells added to pretreatment conditioning regimen on hematopoietic reconstitution after MHC haplotype-mismatched BMT in mice.

METHODSMurine model of MHC haplotype-mismatched BMT was established by using BALB/c(H-2d) x C57BL/6(H-2b) (CB6F(1)(H-2d/b)) mouse as recipient, and C57BL/6(H-2b) mouse as donor. Fifty recipient mice were divided into 5 groups. The mice in the first three groups were each infused 1 x 10(6), 5 x 10(5), 2 x 10(5)/mouse donor-derived NK cells, respectively before TBI ((60)Co, 9.0 Gy) and then conditioned with TBI, followed by infusion of C57BL/6(H-2b) mice bone marrow cells four hours later. The mice in the fourth group received TBI only, and in the fifth group, TBI and BMT at the some doses as the first three groups. Hematopoietic reconstitution, survival time, body weight, histopathology of the recipients were followed up.

RESULTS(1) Survival time was (5.15 +/- 0.66) days for the fourth group, and > 30 days for the other 4 groups. (2) Leukocyte and platelet counts at day 10 after BMT were (0.99 +/- 0.22) x 10(9)/L and (61.0 +/- 7.27) x 10(9)/L respectively for the fifth group and (2.01 +/- 0.21) x 10(9)/L, (101.50 +/- 16.34) x 10(9)/L; (1.98 +/- 0.29) x 10(9)/L, (99.50 +/- 16.41) x 10(9)/L and (1.97 +/- 0.21) x 10(9)/L, (98.0 +/- 16.19) x 10(9)/L for the first three groups, respectively. Histopathology displayed no GVHD in all the groups.

CONCLUSIONDonor-derived NK cells could promote hematopoietic reconstitution after MHC haplotype-mismatched BMT in mice.

Animals ; Bone Marrow Transplantation ; Female ; Graft Survival ; Graft vs Host Disease ; immunology ; prevention & control ; Haplotypes ; Histocompatibility Testing ; Killer Cells, Natural ; cytology ; immunology ; Lymphocyte Transfusion ; Major Histocompatibility Complex ; immunology ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Transplantation Conditioning ; methods ; Transplantation, Homologous

OBJECTIVETo establish an enzyme-linked immunosorbent assay (ELISA) for determining anti-themocyte globulin (ATG) levels in serum samples.

METHODSThe microplate was coated with mouse anti-rabbit IgG monoclonal antibody, and sheep anti-rabbit polyclonal antibody conjugated with HRP was used as the second antibody for detecting the serum ATG levels in patients undergoing allogeneic hematopoietic stem cell transplantation.

RESULTSThe optimal concentration of the coating antibody and dilution ratios of the serum samples and IgG-HRP conjugate were 0.2 microg/ml, 1:40 and 1:2500, respectively. The lower sensitivity limit of the assay was 31.25 ng/ml for ATG detection. A linear relationship was established within the concentration range from 40 to 1000 ng/ml, with the coefficients of variation of 7.91 within assay and 5.22 between assays, respectively. Seven patients undergoing stem cell transplantation with ATG pretreatment showed gradually decreased concentration of ATG, and after 90 days ATG could still be detected.

CONCLUSIONThe sandwich ELISA we established provides a specific and sensitive method for quantitative measurement of ATG in the clinical setting. In patients undergoing stem cell transplantation with ATG pretreatment, the ATG concentration gradually decreases but remains detectable 90 days after the administration.

Adolescent ; Adult ; Antilymphocyte Serum ; blood ; therapeutic use ; Child ; Enzyme-Linked Immunosorbent Assay ; methods ; Female ; Humans ; Leukemia ; blood ; therapy ; Male ; Sensitivity and Specificity ; Stem Cell Transplantation ; Young Adult

Aqueous dispersion and stability of Fe3O4 nanoparticles remain an issue unresolved since aggregation of naked iron nanoparticles in water. In this study, we successfully synthesized different Fe3O4 super-paramagnetic nanoparticles which were modified by three kinds of materials [DSPE-MPEG2000, TiO2 and poly acrylic acid (PAA)] and further detected their characteristics. Transmission electron microscopy (TEM) clearly showed sizes and morphology of the four kinds of nanoparticles. X-ray diffraction (XRD) proved successfully coating of the three kinds of nanoparticles and their structures were maintained. Vibrating sample magnetometer (VSM) verified that their magnetic properties fitted for the super-paramagnetic function. More importantly, the particle size analysis indicated that Fe3O4@PAA had a better size distribution, biocompatibility, stability and dispersion than the other two kinds of nanoparticles. In addition, using CNE2 cells as a model, we found that all nanoparticles were nontoxic. Taken together, our data suggest that Fe3O4@PAA nanoaparticles are superior in the application of biomedical field among the four kinds of Fe3O4 nanoparticles in the future.

Objective:To investigate the expression of Fas、FasL and the apoptosis of liver cancer cell line HepG2 transfected with LIGHT and IFN-? gene mediated by Cationic liposome.Methods:HepG2 cells were divided into two groups(the solo transfection of LIGHT gene and the combined transfection of LIGHT and IFN-? genes) and the control groups(no transfection).HepG2 cells were cellected at 12h,24h and 48h after transfection.The apoptosis of HepG2 cells and the expression of Fas and FasL of the HepG2 cells were investigated with flow cytometry.Results:After transfection,the apoptosis of HepG2 cells increased,and the apoptosis of combined transfection group was higher than the solo transfection of LIGHT(P