Animals ; Arsenic ; toxicity ; Comet Assay ; DNA Damage ; drug effects ; Lymphocytes ; drug effects ; metabolism ; Male ; Malondialdehyde ; metabolism ; Rats ; Rats, Wistar ; Tobacco Smoke Pollution ; adverse effects

OBJECTIVE: To prove the feasibility of detecting menstrual blood as well as its cellular localization with rabbit-anti-human matrix metalloproteinase-11 (MMP-11) polyclonal antibody. METHODS: MMP-11 in menstrual blood, peripheral blood, vaginal liquid, aged menstrual bloodstain, and endometrium sections were assayed with SAP immunohistochemistry. RESULTS: MMP-11 was found only in menstrual samples within stroma and epithelium cells. CONCLUSION MMP-11 polyclonal antibody may be applied in the distinction between menstrual blood and venous blood.
Adult ; Endometrium/pathology* ; Female ; Forensic Medicine/methods* ; Humans ; Immunohistochemistry ; Matrix Metalloproteinase 11/analysis* ; Menstrual Cycle/blood*

Adult ; Chemical and Drug Induced Liver Injury ; Dimethylformamide ; adverse effects ; Formamides ; analysis ; Humans ; Kidney ; physiopathology ; Kidney Diseases ; chemically induced ; physiopathology ; urine ; Kidney Function Tests ; Liver ; physiopathology ; Liver Diseases ; physiopathology ; urine ; Liver Function Tests ; Male ; Middle Aged ; Occupational Exposure

The microbial transformation of buflomedil by Cunninghamella blakesleana AS 3.153 was studied, as well as a microbial model which can be used to mimic metabolism of buflomedil in mammal was established. Experiments were conducted to screen the capabilities of four strains of Cunninghamella species to transform buflomedil, in which C. blakesleana AS 3.153 was selected for a preparative biotransformation. Furthermore, the microbial model was established based on the transformation condition optimization. The parent drug and its metabolites produced by C. blakesleana AS 3.153 were detected by liquid chromatography-mass spectrometry method and three metabolites were identified while two of them were new found metabolites. Two major metabolites, para-O-desmethyl buflomedil and 12-C-oxidated buflomedil, were isolated by semi-preparative HPLC. Based on the comparison between different species, the microbial transformation of buflomedil by C. blakesleana AS 3.153 is more similar to the metabolism of buflomedil in human and Beagle dog than that in rat.
Adult ; Animals ; Biotransformation ; Chromatography, High Pressure Liquid ; Cunninghamella ; metabolism ; Dogs ; Female ; Humans ; Male ; Molecular Structure ; Pyrrolidines ; chemistry ; pharmacokinetics ; Rats ; Rats, Wistar ; Spectrometry, Mass, Electrospray Ionization ; Young Adult

Objective To screen the mutation of certain gene of a 10-years-old boy with multiple xanthomas and very high level of cholesterol who could be diagnosed as homozygous familial hypercholesterolemia (FH),to explore the relationship between the genotype and phenotype,and to discuss the molecular pathologic mechanism.Methods The basic information of life styles were asked from the boy and his familial members.The blood was drown to examine the lipid and genes.The boy was examined with electrocardiogram examination,ultrasonography and coronary CT angiography (CTA) to evaluate the degree of atherosclerosis.Peripheral blood DNA of the boy and his parents were extracted by phenol-chloroform method and investigated for mutations of promoter and all 18 exons of low density lipoprotein receptor(LDLR) gene.Screening was carried out by using Touch-down polymerase chain reaction (PCR) and single strand conformation polymorphism(PCR-SSCP),combined with DNA sequence analysis.In addition,the apolipoprotein B100 gene(apoB100) for known mutations (R3500Q) which caused familial defective apoB100 was screened by PCR-DNA sequence analysis.Results 1.The level of cholesterol of his parents were higher than the normal.2.Several clinical manifestations of atherosclerosis were detected from that boy.Increased intima-media thickness and plaques were detected in the common carotid artery.Mitral valve regurgitation was found by echocardiography.Coronary stenosis was confirmed by CTA.3.No mutations R3500Q of apoB100 was observed.4.A homozygous mutation in exon13 of the LDLR gene (D601Y) were identified in the boy and his parents harbour D601Y heterozygous mutation due to a single base pair substitution of G for T in the codon for residue 1864.Conclusions The final diagnosis of the boy with multiple xanthomas was homozygous FH.His disease was caused by D601Y homozygous mutation in exon13 of the LDLR gene inherited from his heterozygous parents.

OBJECTIVE: To establish an accurate, simple, quick, specific and sensitive method for species identification by amplifying 12S rRNA gene with the same reaction system. METHODS: Based on the downloaded 12S rRNA gene sequences of eleven species (human, chicken, duck, goose, pig, rabbit, rat, sheep, bull, dog and goat) from GenBank, a pair of universal primers to eleven species and three pairs of specific primers to human, chicken and duck were designed. The amplicons amplified with universal primers were used for internal controls, and the amplicons amplified with specific primers were used as identification of human, chicken and duck. DNA was extracted from various samples including blood stains, fresh or freezing muscles, heat-treated muscles and hairs. Both single DNA of human, chicken or duck and mixed DNA of any two kinds of them were amplified. RESULTS: The lengths of universal amplicons were about 400 bp. The lengths of specific amplicons were 163 bp for human, 286 bp for chicken, and 374 bp for duck, respectively. No cross amplification was observed, indicating a high specificity of the specific primers. The identification rate was 100% for human, 99% for chicken, and 100% for duck, respectively. The detection sensitivity ranged from 2.5 pg to 200 pg of DNA concentration depending on species, even in mixtures of different species DNA without interference. CONCLUSION The method established could identify different species under the same reaction system.
Animals ; Blood ; Cattle ; DNA/analysis* ; Dogs ; Forensic Genetics ; Hot Temperature ; Humans ; Polymerase Chain Reaction/veterinary* ; Poultry/genetics* ; RNA, Ribosomal/genetics* ; Rabbits ; Rats ; Sheep ; Species Specificity ; Swine

OBJECTIVE: To investigate the application of ITO method and discriminant functions method in full sibling and half sibling identification. METHODS: Five hundred pairs of full siblings (FS), 50 pairs of half siblings (HS) and 500 pairs of unrelated individuals (UR) were genotyped by PowerPlex 16 system. Full sibling index (FSI), half sibling index (HSI) and the FSI:HSI ratio were calculated with ITO method. Allelic matching of each pair of the three groups was compared. The locus numbers of no-allele sharing (x0), half-allele sharing (x1) and two-alleles sharing (x2) were calculated, respectively. The discriminant functions about full-siblings, half-siblings and unrelated individuals (UR) were established by SPSS 13.0 statistical software. RESULTS: (1) Regard FSI > or = 19 or FSI < 1 as the standard of distinguishing full sibling from unrelated individual, the alternate correct percentage was 96.4%. Regard HSI > or = 19 or HSI < 1 as the standard of distinguishing half sibling from unrelated individual, the alternate correct percentage was 85.3%. Regard FSI:HSI > or = 1 or FSI:HSI < 1 as the standard of distinguishing full sibling from half sibling, the alternate correct percentage was 87.5%. (2) Four groups of discriminant functions were established. The alternate correct percentage of these discriminant functions were 84.4%-97.7%, with the highest one in full sibship-unrelated individual group. CONCLUSION Both ITO method and discriminant functions method are efficient in identification of full sibling or half sibling.
Alleles ; Discriminant Analysis ; Forensic Genetics ; Genetic Variation ; Genomic Imprinting/genetics* ; Genotype ; Humans ; Paternity ; Siblings ; Tandem Repeat Sequences/genetics*

OBJECTIVETo observe therapeutic effect of point-through-point acupuncture on cerebellar ataxia after apoplexy and evaluate the safety.

METHODSRandom, parallel control, single blind and multicentral study method was used and 224 cases from 4 hospitals were divided equally into a treatment group and a control group, 112 cases in each group. The treatment group were treated with point-through-point acupuncture and the control group with general needling method. Their symptoms and signs, and the effect on transcranial Doppler's method (TCD) were investigated.

RESULTSThe total effective rate was 93.3% in the treatment group which was better than 77.4% in the control group, with a significant difference between the two groups (P < 0.01), and the point-through-point acupuncture could significantly improve TCD of basilar artery, vertebral artery and posterior inferior cerebellar artery (Vs, Vm, Vd, PI, RI), superior to the control group.

CONCLUSIONThe point-through-point acupuncture has obvious therapeutic effect on cerebellar ataxia after apoplexy and good safety.

Ranolazine and metabolites in dog urine were identified by LC-MS(n). Dog urine samples were collected after ig 30 mg x kg(-1) ranolazine, then the samples were enriched and purified through solid-phase extraction cartridge. The purified samples were analyzed by LC-MS(n). The possible metabolites were discovered by comparing the full scan and SIM chromatograms of the test samples with the corresponding blanks. Seventeen phase I metabolites and fourteen phase II metabolites were identified in dog urine. Three metabolites were identified by comparing with the control article. The metabolites were formed via the following metabolic pathways: O-demethylation, O-dearylation, hydroxylation, N-dealkylation, amide hydrolysis, glucuronidation and sulfation. The LC-MS(n) method is suitable for the rapid identification of drug and its metabolites in biologic samples.
Acetanilides ; administration & dosage ; metabolism ; urine ; Administration, Oral ; Animals ; Chromatography, Liquid ; Dogs ; Female ; Male ; Piperazines ; administration & dosage ; metabolism ; urine ; Ranolazine ; Solid Phase Extraction ; Spectrometry, Mass, Electrospray Ionization ; Tandem Mass Spectrometry

Our previous studies have demonstrated the effects of brain derived neurotrophic factor (BDNF) on promoting proliferation of multiple myeloma (MM) cells and inducing angiogenesis in MM in vitro. To further investigate whether the PI3K/Akt and MEK1/ERK pathway play a role in the BDNF-induced angiogenesis in vitro and to explore the further molecular mechanisms, two ways to establish human myeloma xenograft animal model were developed, their advantages and disadvantages were elucidated. The phosphorylation of AKT and ERK1/2 were detected in human umbilical vein endothelial cells (HUVECs) by Western blot. The angiogenic activity in vitro was evaluated by transwell migration assay and tubule formation assay. Cell proliferation was determined by crystal violet staining. Cell apoptosis was detected by FITC-Annexin-V/PI double staining and flow cytometry. The results showed that the BDNF activated the PI3K/Akt and MEK1/ERK pathway in the time-dependent manner. Ly294002 and PD98059 blocked the activation of Akt and ERK1/2 respond to BDNF. 100 ng/ml BDNF significantly increased HUVEC tube formation, migration and proliferation in vitro at a similar degree of 25 ng/ml VEGF. Furthermore, tube formation of HUVECs toward BDNF was significantly inhibited by 57% and 40% with 20 micromol/L Ly294002 and 20 micromol/L PD98059 treatment, respectively. At the same time, Ly294002 and PD98059 reduced the BDNF-induced migration of HUVECs by 74% and 36%, respectively. While BDNF-induced survival was only blocked by Ly294002 and BDNF-induced proliferation was only inhibited by PD98059. It is concluded that BDNF promotes angiogenesis of HUVECs in vitro. ERK and Akt are two crucial events in BDNF-mediated signal transduction leading to HUVECs angiogenesis by different mechanisms. Moreover, the latter is more important.
Angiogenesis Inducing Agents ; pharmacology ; Brain-Derived Neurotrophic Factor ; pharmacology ; Chromones ; pharmacology ; Endothelial Cells ; drug effects ; metabolism ; Flavonoids ; pharmacology ; Humans ; MAP Kinase Kinase 2 ; genetics ; metabolism ; Mitogen-Activated Protein Kinase 3 ; genetics ; metabolism ; Morpholines ; pharmacology ; Proto-Oncogene Proteins c-akt ; genetics ; metabolism ; Signal Transduction ; Umbilical Veins ; cytology