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 the relationship between genetic polymorphisms in nucleotide excision repair gene excision repair cross complementing group 6(ERCC6),xeroderma pigmentosum group A(XPA) and coal-burning-borne-arsenism in Guizhou Province.Method ERCC6 A3368G,ERCC6 C-6530G and XPA A23G gene polymorphisms were analyzed by polymerase chain reaction restriction fragment length polymorphism technique(PCR-RFLP) of 205 cases which were chosen as patients with arsenism and 187 residents as control group.Results The distributions of ERCC6 A3368G,ERCC6 C-6530G and XPA A23G in the case group were not statistically significant compared with those of the control group(x2 =3.209,2.963,3.335,all P ＞ 0.05); individuals carrying G allelomorphic gene(AG + GG) had a lower risk than individuals carring A allelomorphic gene(ORadj =0.282,95％CI:0.126-0.628,P =0.002); relationship was not found between single genetic polymorphisms of ERCC6 C-6530G,XPA A23G and coal-burning-borne-arsenism; the risk of arsenism was decreased for individuals carrying the following five genotypes combination:ERCC6 A3368G(AG + GG) genotype and ERCC6 C-6530G CC genotype(ORadj =0.287,95％CI:0.087-0.946,P=0.040); ERCC6 A3368G(AG + GG) genotype and ERCC6 C-6530G(CG + GG) genotype (ORadj =0.226,95％CI:0.077-0.661,P =0.007); ERCC6 A3368G(AG + GG) genotype and XPA A23G AA genotype (ORadj =0.150,95％CI:0.038-0.596,P =0.007); ERCC6 A3368G (AG + GG) genotype and XPA A23G(AG + GG) genotype(ORadj =0.325,95％CI:0.118-0.897,P =0.030) ; ERCC6 C6530G (CG + GG) genotype and XPA A23G AA genotype (ORadj =0.397,95％CI:0.162-0.975,P=0.036).Conclusions Individuals carring ERCC6 A3368G (AG + GG) genotype have a low risk of arsenism.There are five genotypes combination of three gene polymorphisms in two genes,ERCC6 and XPA,which may reduce the risk of coal-burning-borne-arsenism.

Objective To screen the mimotopes ofpemphigus vulgaris (PV) antigen, desmoglein3 (Dsg3) with a phage-displayed random nonapeptide library, so as to update the knowledge on the patho-genesis of PV. Methods Recombinant fusion protein of extracellular domain 1-2 (EC1-2) of Dsg3 and glutathione transferase was expressed by E.coli BL21, and used to purify polyclonal autoantibody binding to recombinant EC 1-2 from the sera of patients with PV. Then, selected autoantibody was applied as a ligand for biopanning of a phage-displayed linear random nonapeptide library and circular random nonapeptide library. Monoclonal phages were selected by immunoscreening and tested with ELISA and competitive ELISA. Results After two rounds ofbiopanning, a population ofpeptide-displaying phages binding to autoan- tidody were highly enriched. Sixty individual phage clones selected by immunosereening were further sub-jected to screening with ELISA and competitive ELISA. Finally, three positive phage clones were obtained. As shown by ELISA and competitive ELISA, they reacted with serum from patients with PV but not with that from normal human controls, and blocked the interaction between patients' sera and recombinant fusion protein of EC1-2. Conclusion Three mimotopes closely associated with PV antigen were successfully selected from a phage-displayed random nonapeptide library.

The drug-loading system of DMF (dextran - magnetic layered double hydroxide - fluorouracil) was synthesized by "co-precipitation intercalated assembly - dextran composite in situ - solvent conversion" technology. The crystal-phase characteristic and slow-release performance of DMF were investigated through X-ray diffraction (XRD), infrared spectrum (IR), transmission electron microscopy (TEM), thermogravimetry (TG) and in vitro release experiment. The targeted transshipment and slow-release effect of DMF system were evaluated by in vivo animal experiment. It was showed that the XRD of DMF matched with R-sixtetragonum type layered double hydroxide and Fd-3m cubic type ferrite. IR test demonstrated that the DMF system was a supra-molecular complex consisted of Dextran (DET), magnetic layered double hydroxide (MLDH) and fluorouracil (FU) components. The two-level supra-molecular MLDH-FU presented six-edge lozenge TEM morphology, with layered characteristics. DET on the surface of DMF was capable of protecting the layered structure of MLDH-FU, improving particle dispersion properties, and strengthening the slow-release performance of the drug delivery system. The drug release model of DMF at pH 7.35 of PBS in vitro fit to the zero-order kinetics equation C = 1.1716 x 10(-5) + 4.4626 x 10(-7) t. The drug delivery system DMF could transport drugs principally to in vivo target organs with a local effect, targeted specificity, and excellent circulation transshipment performance. The pharmacokinetic process of DMF presented multi-peak phenomenon with peak attenuation and cyclic growth. The peaks appeared at 0.25, 1, 3, 5 and 9 d separately after dosing intervention. The first peak process of DMF accorded with a pharmacokinetic equation of C(FU) = 14.34 e(-0.530t) + 36.04 e(-0.321t) + 24.18 e(-0.96t), and presented the characteristic of slow absorption and fast elimination. As for subsequent peak processes, half-life increased, bioavailability increased, and plasma clearance decreased. The highest peak value of DMF was 1/37 of original value of FU, and the relative bioavailability was 419% to original FU.

By using rice SSRP, RAPD and AFLP molecular markers, the genome of rice transgenic line "Minghui 63-Xa21" was analyzed. 32 SSRP primers, 42 RAPD primers and 8 AFLP primers could produce obvious PCR bands in the analysis of at least 12 individual plants selected randomly from "Minghui 63-Xa21" T3 generation. Totally 550 PCR bands, equivalent to 550 genomic sites, were detected. Different individual plants of the transgenic homozygous line displayed almost the same PCR pattern. Compared with the control "Minghui 63", no difference was found in their PCR patterns. This indicated that the introduction of Xa21 into the genome of "Minghui 63" did not change these 550 genome sites and their heredity. Very few variant PCR bands were observed in some individual plants from both "Minghui 63-Xa21" and "Minghui 63". However, the variant percentage was equivalent between the transgenic line and the non-transgenic control line.
Chromosome Mapping ; methods ; Genome, Plant ; Microsatellite Repeats ; genetics ; Oryza ; genetics ; Plant Proteins ; genetics ; Plants, Genetically Modified ; Protein-Serine-Threonine Kinases ; genetics ; Random Amplified Polymorphic DNA Technique ; methods

To investigate the effects of carbamazepine (CBZ) on the plasma concentrations of valproic acid (VPA) and its toxic metabolite 2-propyl-4-pentenoic acid (4-ene VPA) in epileptic patients, the plasma concentrations of VPA and 4-ene VPA were determined, and the effect of CBZ on pharmacokinetics of VPA was evaluated. All patients had been divided into two groups (VPA group, n = 87; and VPA+CBZ group, n = 19). As compared to VPA group, the combination of CBZ significantly (P < 0.01) decreased the trough concentration of VPA [VPA group, (69.5 +/- 28.8) microg x mL(-1); VPA+CBZ group, (46.3 +/- 25.6) microg x mL(-1)] and does-adjusted VPA trough concentration [VPA group, (4.89 +/- 2.21) microg x mL(-1) x mg(-1) x kg(-1); VPA+CBZ group, (3.14 +/- 1.74) microg x mL(-1) x mg(-1) x kg(-1)]. However, the addition of CBZ did not influence the concentration of 4-ene VPA. The present study revealed that coadministration of CBZ can reduce VPA plasma concentration and may impact VPA clinical effect, therefore therapeutic drug mornitoring of VPA should be used when combined use of CBZ and VPA.
Adolescent ; Adult ; Anticonvulsants ; blood ; pharmacokinetics ; therapeutic use ; Carbamazepine ; blood ; pharmacokinetics ; therapeutic use ; Drug Interactions ; Drug Therapy, Combination ; Epilepsy ; blood ; drug therapy ; Fatty Acids, Monounsaturated ; blood ; Female ; Humans ; Male ; Valproic Acid ; blood ; pharmacokinetics ; therapeutic use ; Young Adult

OBJECTIVETo explore the clinical effects of external fixation associated with limited internal fixation on treatment of Gustilo grade III leg fractures.

METHODSFrom July 2006 to December 2008, 40 cases of Gustilo grade III leg fractures were emergently treated in our unit with external fixation frames. Soft tissue injuries were grouped according to the Gustilo classification as IIIA in 17 cases, IIIB in 13 cases, and IIIC in 10 cases. All the patients were debrided within 8 hours, and then fracture reposition was preformed to reestablish the leg alignment. Limited internal fixation with plates and screws were performed on all the Gustilo IIIA cases and 10 Gustilo IIIB cases at the first operation. But all the Gustilo IIIC cases and 3 Gustilo IIIB cases who had severe soft tissue injuries and bone loss only received Vacuum-sealing drainage (VSD). Broad-spectrum antibiotics were regularly used and VSD must be especially maintained easy and smooth for one week or more after operation. Limited internal fixation and transplanted free skin flaps or adjacent musculocutaneous flaps were not used to close wounds until the conditions of the wounds had been improved.

RESULTSThe first operations were completed within 90-210 minutes (170 minutes on average). The blood transfusions were from 400 ml to 1500 ml (those used for anti-shock preoperatively not included). All the 40 patients in this study were followed up for 6-28 months, 20.5 months on average. The lower limb function was evaluated according to the comprehensive evaluation standards of leg function one year after operation and the results of 28 cases were excellent, 9 were good and 3 were poor.

CONCLUSIONExternal fixation associated with limited internal fixation to treat Gustilo grade III leg fractures can get satisfactory early clinical therapeutic effects.

Adolescent ; Adult ; Debridement ; External Fixators ; Female ; Fracture Fixation, Internal ; methods ; Humans ; Leg Injuries ; surgery ; Male ; Middle Aged

Traditional Chinese medicine (TCM) property theory is believed to be a key and difficult point of basic theory studies of TCM. Complex concepts, components and characteristics of TCM property have long puzzled researchers and urged them to develop new angles and approaches. In the view of cognitive science, TCM property theory is a cognitive process of storing, extracting, rebuilding and summarizing the sensory information about TCMs and their effects during the medical practice struggling against diseases under the guidance of traditional Chinese philosophical thinking. The cognitive process of TCM property has particular cognitive elements and strategies. Taking into account clinical application characteristics of TCMs, this study defines the particular cognitive elements. In the combination of research methods of modern chemistry, biology and mathematics, and on the basis early-stage work for five years, we have built a TCM property cognition model based on three elements and practiced with drugs with pungent and hot properties as example, in the hope of interpreting TCM properties with modern science and providing thoughts for the nature of medical properties and instruction for rational clinical prescription.
Animals ; Cognition ; Drug Therapy ; psychology ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Humans ; Medicine, Chinese Traditional ; psychology ; Models, Theoretical

OBJECTIVETo explore the expression and clinical significance of Semaphorin4D (Sema4D) mRNA in peripheral blood lymphocyte, Sema4D on platelet surface, soluble Sema4D (sSema4D) in plasma in patients with cerebral infarction.

METHODSTaking 299 patients with cerebral infarction as the case group while 195 healthy adults as the control group. The mRNA expression of Sema4D was detected by Real-time PCR, and Sema4D expression on platelet by flow cytometry, sSema4D by ELISA. Then, the expression of Sema4D on platelet surface and the concentration of sSema4D in plasma of the 195 selected patients following 2 weeks' treatment were tested.

RESULTSThe expression of Sema4D mRNA significantly increased in the case group \[(2.23, 2.66)×10(4) IU/ml\] than in the control group \[(0.49, 0.53)×10(4)IU/ml\] (P < 0.01). The level of Sema4D on platelet surface in the case group (191.62 ± 46.56) significantly decreased than in the control group (303.33 ± 112.66) (P < 0.01). But the concentration of sSema4D in plasma in the case group \[(1.34 ± 0.56) µg/L\] was obviously higher than in the control group \[(0.61 ± 0.31) µg/L\] (P < 0.01). The expression of Sema4D on platelet was obviously relevant with the concentration of sSema4D in plasma in the case group with the correlation coefficient as 0.328 (P < 0.01). The expression of Sema4D on platelet obviously peaked up following 2 weeks' routine therapy in the case group, which was close to that in the control group. Meanwhile the concentration of sSema4D in plasma was downward corrected to the normal in the case group.

CONCLUSIONThe increased expressions and plasma levels, and reduced expressions on platelet of Sema4D in acute period, which returned to normal 2 weeks after treatment in the case group may be related to the occurrence of acute cerebral infarction, reflecting the development process of cerebral infarction.

Aged ; Antigens, CD ; blood ; metabolism ; Blood Platelets ; metabolism ; Case-Control Studies ; Cerebral Infarction ; blood ; Female ; Humans ; Lymphocytes ; metabolism ; Male ; Middle Aged ; Semaphorins ; blood ; metabolism

OBJECTIVETo evaluate the expression and activity of TNF-related apoptosis-inducing ligand (TRAIL) gene expressed from the hTERT promoter on colon cancer cell line HT-29.

METHODSGFP/TRAIL gene expressed from the hTERT promoter was transfected into HT-29 with adenoviral vectors system, expression and apoptosis inducing ability of GFP/TRAIL protein were determined with fluorescence-activated cell sorting (FACS) method.

RESULTSThe expression of GFP gene was 31.4 % and 67.0 % with either hTERT promoter or CMV promoter in DLD1 cells; GFP/TRAIL gene was able to inhibit cell growth (74.2%) and induce apoptosis (25.8%) of HT-29 cells. There was significant difference between Ad/hTERT-gTRAIL and the other two control groups (PBS and Ad/CMV-GFP, P<0.05).

CONCLUSIONThe GFP/TRAIL gene with hTERT promoter transfected by adenoviral vector was successfully expressed in HT-29 cell, which can both inhibit cell growth and induce apoptosis of colon cancer cell line HT-29.

Adenoviridae ; genetics ; Apoptosis ; physiology ; Colonic Neoplasms ; enzymology ; pathology ; Genetic Vectors ; Green Fluorescent Proteins ; biosynthesis ; genetics ; HT29 Cells ; Humans ; Promoter Regions, Genetic ; genetics ; TNF-Related Apoptosis-Inducing Ligand ; biosynthesis ; genetics ; Telomerase ; genetics ; Tumor Cells, Cultured