Chromatography, High Pressure Liquid ; methods ; Humans ; Neonicotinoids ; Pyridines ; blood

Objective Overexpression of multidrug resistance(MDR)is one of the mechanisms that will lead to chemotherapy failure.Of the MDR pathways in tumor cells,P-glycoprotein (P-gp) encoded by MDR genes is one of the key points.99Tcm-methoxyisobutylisonitrile(MIBI) is an imaging agent that can detect overexpression of MDR in tumor cells.The aim of this study was to observe the relationship between 99Tcm.MIBI uptake kinetics and P-gp levels in tumor cells,with or without MDR reversing agents.Methods A totle of 2×106 cells of human myelogeneous leukemia cell line K562 and its resistant subline(K562/D) were incubated with 8 MBq 99Tcm-MIBI respectively.99Tcm-MIBl accumulation and emux at various time inter-vats and the uptake difference with the presence of different cyclosporin A(O.1-O.4 ug/ml)were also ob-served.Comparison of different cell lines or without and with cyclosporin A were performed with the t-test, and the daa of different groups were compared by q-test.Results 99Tcm-MIBI uptake in K562 and K562/D cell line were 1.559±0.529 and 0.107±0.036,99Tcm-MIBI uptake in k562 was flitleon times higher than k562/D.As compared with K562 cell line with no expression of P-gp,significantly increased the 99Tcm-MIBI uptake of K562/D to 106%,148% and 163%after treated with cyclosporin A(0.1,0.2.0.4ug/ml)was ob-served(t=4.35,4.83,5.88,P<0.05).Conclusiom 99Tcm-MIBI uptake can reflect the P-gP level and multidrug-resistance inhibitor may impact 99Tcm-MIBI uptake in P-gP overexpressing cells.

The violence behavior is a kind of aggressive behaviors or attempt to hurt another person psychologically,physically or in other forms.Recently,violent incidents occur more and more frequently,and especially among teenagers.A number of concerns on violence continue to rise,and interpersonal violence is the most concerned type.Perpetrators are violence implementers who determine the occurrence and outcome of violence.Many studies provided the risk factors of interpersonal violence,that expounds the influence of personal level,interpersonal relationship,community background and social factors of interpersonal violence.The establishment of the socioecological risk-factor structural model which focuses on the perpetrators' indi vidual,is of great significance for the effective intervention for interpersonal violence.

Objective:Aldose reductase,involved in the pathogenesis of diabetic complications,was recombinated with an adeno associated virus vector pSNAV2.0,and it was transfected into human embryonic kidney 293(HEK 293)cells.The gene engineering produced AR would be used as a target protein to screen aldose reductase inhibitors.Restriction endonuclease digestion and ligation procedures were performed to construct the AR expression plasmid vector pSNAV-hAR.Methods:After confirmation the recombinant plasmid by PCR,restriction endonuclease digestion,and DNA sequencing,pSNAV-hAR was transfected into HEK293 cells.Western blot and immunofluorescence analysis were performed to detect the expression of AR and its enzyme activity.Results:The results of a series of analysis including AR activity assay,Western blot and immunofluorescence analysis shown the expressed protein mediated by the adeno associated virus vector transfecting HEK 293 cells,was functional AR.The traditional aldose reductase inhibitors,Sobinil and Zopolrestat,were used to test and verify the constructed cell model.Conclusion:The established AR expression model can be used in mechanismresearch of activation of polyol pathway on diabetic complications and screening potential aldose reductase inhibitors.

Background Allograft rejection is a main cause of failure of penetrating keratoplasty,especially in the patient with high risk of rejection condition.Previous study on allograft rejection mechanism focused on limbal and corneal neovascularization,but these factors did not explain all the phenomena of allograft rejection.Research found that immune cells appeared in iris and ciliary body when rejection occurred,but the relationship between these immune cells and allograft rejection is unclear Objective This study was to evaluate the relationship between diversity of vascular permeability in the iris and ciliary body and allograft rejection after penetrating keratoplasty.Methods Seventy clean eight-week-old BALB/c mice were divided into allogeneic corneal transplantation group (60 mice) and blank control group (10 mice).Allogeneic corneal transplantation was performed with the same age of C57BL/6 mice as donor and BALB/c mice as the recipients.The grafts were examined under the slit lamp microscope and scored based on the criteria of Hegde.The mice were sacrificed and iris and ciliary tissue were obtained 5,10 days and rejection after surgery.Immunohistochemistry and reverse transcription PCR (RT-PCR) was used respectively to detect the expression diversities of occludin,zonula occludens protein-1 (ZO-1),matrix metalloproteinase-9(MMP-9),major histocompatibility complex-Ⅱ (MHC-Ⅱ),and CCR5,CCR7 and their mRNA in iris and ciliary body.Image-J image analysis software was used to calculate the quantity of positive cells on iris wholemount,and absorbance of target genes (A values).The use and care of the experimental animals complied the ARVO Resolution on the Use of Animals in Research.Results The mean survival time of corneal gratts was (17±3) days after operation.The mean score was 0.6 in 5 days and 0.5 in 10 days,and 3.3 in 18 days after operation.Expression of ZO-1 reduced significantly,and that of MMP-9 increased obviously at the time of rejection.MHC Ⅱ + cells were scattered in iris and ciliary body in normal mice,and the number of the positive cells (cells/field) was increased after operation with a peak value when rejection occurred.A significant difference was seen between normal mice and rejection mice (1559.67±350.29 vs.4021.83±495.18) (P=0.000).The expressions of occludin mRNA and ZO-1 mRNA in the iris and ciliary body decreased obviously in the rejection mice.Compared with normal mice,theA value of ZO-1 and occluding were 36.74±3.13 vs.110.11±11.88 and 57.54±3.41 vs.59.90±3.50respectively,with significant differences between them (all P＜0.05).The expressions of MMP-9 mRNA,CCR5 mRNA and CCR7 mRNA in the iris and ciliary body increased gradually with the time lapse after operation and peaked when the rejection appeared.The A value of MMP-9 mRNA,CCR5 mRNA and CCR7 mRNA were significantly higher than those of normal mice (20.29±1.19 vs.2.77±0.85 for MMP-9 mRNA; 35.43±2.56 vs.9.11±0.29 for CCR5 mRNA,and 60.83±0.87 vs.0.89 ±0.95 for CCR7 mRNA) respectively (all P＜0.05).Conclusions The permeability of vascules in the iris and ciliary body increase during the allograft rejection after penetrating keratoplasty.Increased antigen presenting cells were also detected.

Photoaffinity labeling is widely applied to demonstrate targets of small molecule ligands. In this paper, biotin photoaffinity labeled molecule with propargyl group 1 has been designed and synthesized, followed it's labeling of N2-acetyl-2'-O-propargyl guanosine 9 by "click chemistry". This technology presents delight development potential in labeling of second messenger cyclic nucleotide, antisense oligonucleotide or siRNA.
Biotin ; chemistry ; Click Chemistry ; Guanosine ; chemical synthesis ; chemistry ; Ligands ; Photoaffinity Labels

The study reports the detection of neuroprotective effect of 10 kinds of coumarin derivatives and explores their possible mechanism. MTT method was used to screen the neuroprotective effect of 10 coumarin derivatives on neurotoxic agents (Aβ25-35 and rotenone) or OGD (oxygen-glucose deprivation). A compound with better protective effect was obtained. Then the effect of this compound on neurotoxic agents on PC12 was detected by the morphological observation. Furthermore, the effect of compound 3 on microglia with lipopolysaccharide (LPS) induced inflammation was detected. And the inflammatory factor was tested. Finally, direct free radical scavenging ability was detected. Compound 3 was found to be the best compound through three neurons toxic models. Not only compound 3 ameliorated cell viability reduced by three neurons toxic models, but also significantly inhibited the production of inflammatory factor (TNF-α and IL-1β). And its free radical scavenging ability is very good, especially the effect on superoxide anion, which is comparable with vitamin C. The significant scavenging effect of compound 3 on superoxide anion might be the mechanism of the neuroprotection. Compound 3 as a potential neural cell protective agent merits further investigation.
Animals ; Coumarins ; chemistry ; Free Radical Scavengers ; chemistry ; Inflammation ; Microglia ; drug effects ; Neurons ; drug effects ; Neuroprotective Agents ; chemistry ; PC12 Cells ; Rats

Objective:To find out whether it is accurate to estimate femoral version based on femoral broach after femoral neck osteotomy using computed tomography scans.Methods:In 32 total hip arthro-plasty (THA),we performed CT scans before and after operation.Four possible levels (lesser trochan-ter,5 mm above,10 mm above and 15 mm above the lesser trochanter)of broach version were calculated based on the pre-operative CT scan.Stem versions were measured on the post-operative CT scan.We de-termined the difference between the preoperative broach version and the postoperative stem version using the Student’s t-test for paired samples assuming equal variance.Results:For the operated hips,pre-operative hip version differed according to the level of measurement.Our findings showed that the average femoral version was 37.0°±11.0°at the level of the lesser trochanter (section 1),34.3°±10.6°at 5 mm above the lesser trochanter (section 2),28.1°±10.9°at 10 mm above the lesser trochanter (sec-tion 3),and 22.4°±13.7°at 15 mm above the lesser trochanter (section 4),and that the average ver-sion for the femoral neck (FNV)was 12.9°±13.8°.The postoperative hip version was the stem version (FSV),which we found to be an average of 26.1°±11.0°.The mean femoral version for section 1 and 2 was larger than the mean postoperative stem version (P<0.01);the mean version for sections 3 and 4 did not differ from the mean postoperative stem version (P>0.05).The mean femoral neck version was less than the mean postoperative stem version (P<0.01);the difference was 13.2°±11.1°of the in-creased anteversion on average for the FSV compared with FNV.Conclusion:The accuracy of estimated femoral version after arthroplasty depends on broach level.When it is 10 mm above the lesser trochanter, stem version estimation is accurate,but below that level,there is a tendency to overestimate.

Embryonic callus of Kentucky bluegrass were used as material for genetic transformation with the DREB1A gene by particle bombardment. Parameters of biolistic bombardment were studied. The optimal methods showed as follows: plasmid DNA are coated by Ca(NO_3)_2 and PEG4000, 1?m golds are the vectors of plasmid DNA, 6 cm bombardment height distance, 1 time and without osmotic treatment are favorable to transgenic efficiency. The concentration hygromycin (Hy) , which is the selection mark for cultivar‘Baron’ is 100mg/L.

OBJECTIVE To explore the effects of perinatal inflammation on the expression of proin-flammatory cytokines and DMEs (drug metabolism enzymes) in offspring mice. METHODS C57BL/6 maternal mice were administrated with single dose 100 μg·kg-1LPS(lipopolysaccharide)or saline(vehicle) during gestation (day 10 after fertilization). Offspring mice were sacrificed at 30 d after birth and liver samples were collected.Real-time PCR was adopted to test the mRNA expression of proinflammatory cytokines (Nrlp3 and IL-1β), nuclear receptors (Pxr and Car), and DMEs (Cyp3a11, 2b10, 1a2, and Ugt1a1).RESULTS Gender different expression of candidate genes was observed.The expression of Car,in the maternal injection of LPS groups,was significantly decreased in both female and male offspring (n=3-8/group, P<0.01). Concomitantly, a significantly lower expression of Cyp3a11 was found in both female and male offspring (P<0.01, P<0.05, respectively). Furthermore, the expression of Ugt1a1 was reduced in male offspring following maternal administration of LPS (P<0.01). In male offspring, Nrlp3 expression was specially decreased(P<0.05).Interestingly,there was an approximately 66% reduction in mRNA level of Cyp1a2 in female offspring (P<0.01), while in male offspring Cyp1a2 expression showed an increased trend (P>0.05) compared with vehicle group. The expression of Pxr, Cyp2b10, and IL-1β was no difference between LPS treatment group and vehicle group(P>0.05).CONCLUSION Maternal LPS administration affects the expression of proinflammatory cytokines, nuclear receptors and DMEs in mouse offspring.