Objective T o infer the frequency of dosage and m edication history investigate of the victim s in drug facilitated cases by the segm ental analysis of clonazepam in hair. Methods Freezing m illing un-der liquid nitrogen environm ent com bined w ith ultrasonic bath w as used as sam ple pretreatm ent in this study, and liquid chrom atography-tandem m ass spectrom etry w as used for the segm ental analysis of the hair sam ples collected from 6 victim s in different cases. T he concentrations of clonazepam and 7-am in-oclonazepam w ere detected in each hair section. Results C lonazepam and its m etabolite 7-am inoclon-azepam w ere detected in parts of hair sections from the 6 victim s. T he occurrence tim e of drug peak concentration w as consistent w ith the intake tim ing provided by victim s. Conclusion Segm ental analysis of hair can provide the inform ation of frequency of dosage and intake tim ing, w hich show s an unique evidential value in drug facilitated crim es.

Objective T o establish a gas chrom atography-m ass spectrom etry (G C-M S ) m ethod for the determ ination of sulfide ion in blood and apply it to the practical cases. Methods T he 1, 3, 5-tribro-m obenzene w as selected as an internal standard, and 0.2 m L blood sam ple w as collected and analyzed using G C-M S after α-B rom o-2, 3, 4, 5, 6-pentafluorobenzyl brom ide derivatization. Results T he m ass concentration of sulfide ion in blood had good linearity in the range of 0.2-40μg/m L w ith a lim it of detection (L O D ) of 0.05μg/m L . T he m ass concentration of sulfide ion w as less than 0.05μg/m L in blank blood from different sources such as healthy subjects and dead cases. In 3 sulfide poisoning cases, sul-fide ion w as detected in the blood sam ples of 6 victim s, and the m ass concentration range w as 1.02-3.13μg/m L . Conclusion T his study establishes a m ethod for investigation of sulfide ion in blood w hich has been applied successfully to the cases of fatal sulfide poisonings.

Objective To evaluate tear ferning changes of conjunctivochalasis.Design Prospective case study series.Partici- pants 30 patients(60 eyes)of conjunctivochalasis and normal subjects were selected.Methods The subjects were observed with gen- eral ophthalmic examination and tear fern test(TFT).Tear ferning was classified into 4 types.TypeⅠand TypeⅡare normal.TypeⅢand TypeⅣare abnormal.Main Outcome Measures The type of tear feming.Results TFT showed that tear ferning was de- creased in conjunctivochalasis group(TypeⅢand TypeⅣoccupied 61.7%).The difference between conjunctivoehalasis and normal control group was significant(P

As a keratinized material, nail recently has attracting researchers' attention in the pharmaceuticals analysis. There are comparatively limited studies concerning nail's xenobiotic determination and its mechanism. This article reported the development of a sensitive, specific and reproducible LC-MS/MS method, which could be as a foundation of other studies on drug determination in nail. It can also be regarded as the first report on organic drug in mainland China. Sixteen nail samples from volunteers, who were ingested clozapine for more than nine months, are confirmed positive after being analyzed by the method. It is found that contents of clozapine in the patients' nails are above the nanogram level. Besides, a comparative study of clozapine concentration in nails and hair was made, with a result that there exists a correlation between the two materials in terms of clozapine concentration.
Adult ; Antipsychotic Agents ; pharmacokinetics ; China ; Chromatography, Liquid ; methods ; Clozapine ; pharmacokinetics ; Female ; Hair ; chemistry ; Humans ; Male ; Middle Aged ; Nails ; chemistry ; Psychotic Disorders ; metabolism ; Tandem Mass Spectrometry ; methods

OBJECTIVETo observe the effects of AML1-ETO fusion gene on the transcription activity of p21WAF1/CIP1 gene. And to explore the enhancement of leukemia pathogenesis of AML1-ETO.

METHODSThe luciferase reporter plasmids of p21WAF1/CIP1 gene promoter were constructed, and co-transfected into CV-1 cells with AML1-ETO, AML1b and AML1a expression plasmids. The trans-activity of p21WAF1/CIP1 gene promoter was assayed by luminometer.

RESULTSAML1-ETO exhibited a distinct inhibition activity of p21WAF1/CIP1 gene promoter with a sequence-specificity and dosage-dependent manner. The trans-activity of p21WAF1/CIP1 gene promoter decreased to (19 +/- 4)% compared to control group, when 1000 ng pCMV5-AML1-ETO plasmid was used. AML1b and AMLla showed less inhibition activity. The trans-activity of p21WAF1/CIP1 gene promoter decreased to (61 +/- 16)% and (59 +/- 16)% compared to control group, respectively, when 1000 ng plasmid was used.

CONCLUSIONAML1-ETO exhibits more inhibition activity of p21WAF1/CIP1 gene promoter than AML1b and AMLla, results from recruiting transcription co-repression complex efficiently by ETO. Based on previous researches, the effects of exogenous AML1-ETO on p21WAF1/CIP1 gene promote may be dependent on the type of cell lines.

Animals ; Cell Line ; Core Binding Factor Alpha 2 Subunit ; genetics ; Cyclin-Dependent Kinase Inhibitor p21 ; genetics ; Haplorhini ; Oncogene Proteins, Fusion ; genetics ; Plasmids ; genetics ; Promoter Regions, Genetic ; genetics ; RUNX1 Translocation Partner 1 Protein ; Transcription, Genetic ; Transfection

Acupuncture Therapy ; methods ; Adult ; Female ; Humans ; Male ; Middle Aged ; Nerve Compression Syndromes ; therapy ; Skin ; innervation ; Upper Extremity ; innervation

OBJECTIVETo explore the effects of beta-elemene combined with aclarubicin on the induction of HL-60 cell apoptosis and its mechanisms in antileukemia therapy.

METHODSHL-60 cells were treated for 20 hours with different dose of aclarubicin (0.05, 0.10, 0.25 microg/ml) or with different concentrations of beta-elemene (10, 20, 40 microg/ml) in the presence or absence of aclarubicin (0.10 microg/m). The apoptotic rate was analyzed by flow cytometry (FCM), the productions of PGE2 in culture supernatants was detected by competitive ELISA and the expressions of COX-2 and NF-kappaB activity in HL-60 cells by Western blot.

RESULTSLower concentration of aclarubicin (0.05, 0.10 microg/ml) didn't affect apoptotic rate, and COX-2, NF-kappa B and PGE2 expression on HL-60 cells. Combined treatment of beta-elemene and aclarubicin (0.10 microg/ml) enhanced the apoptotic effect and down-regulated COX-2, NF-kappaB and PGE2 expressions. There was a positive correlation between the effects and beta-elemene concentrations.

CONCLUSIONbeta-elemene enhances aclarubicin-mediated apoptotic effect, down-regulation of COX-2 and their inducing products PGE2 in HL-60 cells by suppressing activitation of NF-kappaB.

Aclarubicin ; Apoptosis ; drug effects ; Cell Line, Tumor ; Down-Regulation ; HL-60 Cells ; Humans ; NF-kappa B ; metabolism

OBJECTIVETo study the proto-oncogene RON mediated aggression of Raji cells and the inhibitory effects by monoclonal antibody Zt/f2 (2f2).

METHODSThe effects of RON ligand macrophage stimulating protein (MSP) (2.0 nmol/L) and inhibitory Zt/f2 (2F2) (2.0 nmol/L) antibody on proliferation of RON positive Raji cells after treatment for 24 and72 hours were detected by MTT method, colony formation units (CFU) of Raji cells by methylcellulose semi solid culture, Raji cells apoptosis and cell cycle analysis by AnnexinV/PI double staining, expression of RON, apoptosis-related proteins, and cyclins by Western blot.

RESULTS(1)Compared with the cell viability (1.0) and counts of CFU (103.6±7.0) in control group, Raji cells after MSP treatment had better viability (1.35±0.20) and CFU counts (133.7±10.4) (P<0.05), but worse viability (0.68±0.11) and CFU counts (66.3±6.1) after Zt/f2 (2F2) treatment (P<0.05). (2)Percentage of Raji cells apoptosis after Zt/f2 (2F2) antibody treatment (12.16±2.33)% was significantly increased than the control (2.89±1.03)% (P<0.05). The percentage of Raji cells arrested in G0/G1 phase was increased after Zt/f2 (2F2) antibody treatment as compared to the control [ (54.96 ±3.70)% vs (39.10±2.30)%, (P<0.05) ]. (3) High-level of RON phosphorylation and β-catenin expression activated by MSP could be inhibited significantly by Zt/f2 (2F2), which also up-regulated the expression of caspase-3, caspase-8, caspase-9 and PARP and down-regulated anti-apoptotic MCL-1 gene and inhibitor of apoptosis protein XIAP expression, accompanied with G1 phase protein changes accordingly.

CONCLUSIONMSP could aggravate Raji cells proliferation. Inversely, Zt/f2 (2F2) could inhibit proliferation and induce apoptosis by inhibition of RON phosphorylation and up-regulation of apoptosis related proteins.

Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Humans ; Proto-Oncogenes ; Receptor Protein-Tyrosine Kinases ; metabolism

OBJECTIVETo study the spatial distribution of free Ca2+ in osteoclast-like cells cultured on glass.

METHODSTo detect the free Ca2+ in osteoclast-like cells, the images were analyzed with image software, using the laser scanning confocal microscope and fluorescent probe.

RESULTSAt 37 degrees C the free Ca2+ in osteoclast-like cells could be labelled effectively with 10 micromol/L Fluo-3/AM, the intensity of Ca2+ fluorescent signal in the central part was greater than that in the peripheral part and in the same section the signal was not distributed evenly.

CONCLUSIONThe intensity of Ca2+ fluorescent signal is different among various organellae in osteoclast-like cell, which suggests the osteoclast-like cell modulate its own function through the spatial difference of free Ca2+ concentration.

Aniline Compounds ; Calcium ; Cell Line ; Cells, Cultured ; Microscopy, Confocal ; Osteoclasts ; Xanthenes

Objective To observe apelin and its receptor (APJ) expressions in human osteoblasts and evaluate the effect of apelin on osteoblasts.Methods The expressions of apelin and APJ in human osteoblasts were tested by RT-PCR and Western blot.After human osteoblasts were treated with apelin,cell proliferation was measured by [~3H] thymidine incorporation and cell counting.Cell function was measured by alkaline phosphatase (ALP) activity,the secreted osteocalcin level and typeⅠcollagen production .The activation of signaling cascades was tested by Western blot.Small-interfering RNA (siRNA) to blockade APJ was applied to observe effects of apelin on cell proliferation and the activation of signaling cascades.Results Both apelin and APJ were expressed in human osteoblasts.Apelin increased the proliferation and did not show the influences on ALP activity, osteocalcin secretion and type I collagen production in human osteoblasts.Apelin induced activation of phosphatidylinositol-3 kinase (PI3K) downstream effector (Akt),but not mitogen-activated protein kinase (MAPK) such as c-jun N-terminal kinase (JNK),p38 and ERK1/2 in human osteoblasts.Suppression of APJ with siRNA or LY294002 (PI3K inhibitor) abolished the apelin-induced cell proliferation and the activation of Akt.Conclusion Human osteoblasts express apelin and APJ.Apelin stimulates the proliferation of human osteoblast via APJ/PI3K/Akt pathway,but has no effect on osteoblast differentiation.