Objective To screen the microsatellites with low occurrence rate of stutter band and establish the effective bovine STR typing system.Methods The tetranucleotide STR loci in bovine genome were searched with Tandem Repeat Finder software.Primers were designed and used to amplify these candidate loci and the PCR products were separated with electrophoresis.DNA samples from 100 head of unrelated cattle were typed.Results Among these candidate loci,6 bovine tetranucleotide STR loci showed high polymorphism,and their CDP and CPE value were 0.99995 and 0.859591 respectively.Conclusion The 6 bovine tetranucleotide STR loci can be used for bovine identification and parentage testing.

Oncogene c-jun is a member of jun family,the immediately early genes(IEGs),and belongs to one of the nuclear transcription factors of basic leucine zipper(bZIP)family.Combined with many gene promotors,c-jun is involved in the regulation of gene transcription.Its products play important roles in regulating gene expression,cell proliferation,differentiation and apoptosis.The structure,biological funetion,regulation of c-jun and its roles contributing to tissue damage are reviewed in this article,which may provide understanding for severity of tissue injury and wound age estimation in the field of forensic pathology.

Aconltium plants are a group of important poisonous plants in the Ranunculaeeae family of Angiosperm subphylum.and one of the earliest recorded poisonous plants in China as well.In their tubers(root)have hypertoxic Aconitium alkaloids,which belong to the group of diesterditerpene alkaloids.It is repeatedly reported in China that aconitine poisoning,and even death from aconitine poisoning,caused by the individual differences in pharmaceutical tolerance of aconitine,taking the wrong medicine or inadequate dosage,or put in the poison by homicide.At present,the research on aconitine is mainly limited to clinical treatment,but lack of research on toxicity mechanism of aeonitine.This paper reviews some progress relating to the toxicological mechanisms of this kind of alkaloid.

Objective To evaluate the probability of siblings identification in Identifiler system by using the software of automatic analysis.Methods Using the software of automatic analysis in siblings jdentification.STP genetic typing of 151 pairs of full siblings and 31224 pairs of unrelated individuals from manual simulation were analyzed in 15 STR loci of ldentifiler system.Results Kin probability(W_(FS))of 39.07% full siblings were more than 99.999% while W_(FS) of unrelated individual pairs were 0% .W_(FS) of 60.93% full siblings and 21.3% unrelated individual pairs were all at the range from 99.999% to 1% .W_(FS) of 78.7% unrelated individual pairs 0% full siblings individuals were less than 1% .Therefore,there were notability difference between full siblings and unrelated individual pairs.In addition,testing of 15 STR loci of Identifiler system,it suggested that the pair were siblings when the locus number of the entirely-same is not less than 5 or that of the entirely-different is not more than 1,and that the pair were unrelated individuals when the locus number of the entirely-different is not less than 6 or that of the entirely-8alne is not more than 1.Conclusion The software of automatic analysis and the Identifiler system call be used to siblings identification.

Objective To study the relationship between the DNA content of chondrocytes in the costal cartilage and postmortem interval in putrefactive rat cadavers.Methods Nuclear DNA wag visualized by modified Feulgen's staining method.DNA content of ehondrocytes in the costal cartilage was semi-quantita tively determined by a computerized image analysis system in rats within 35d postmortem.Results Staining intensity of the nuclei was gradually reduced within from 1d to 28d postmortem.The nuclej could not be detected at 35d.The DNA content of chondrocytes decreased time-dependently within 28 days after death as determined semi-quantitatively,which revealed a linear relationship between DNA content and postmortem interval.Conclusion DNA content of chondrocytes in the costal cartilage reduces time-dependently with the extension of postmortem interval.

Objective To study the application of a new magnetic beads made in China in DNA extraction of forensic biological samples with automation workstation.Methods DNA was extracted from common forensic biological samples by QIAGEN Bio-Robert Universal System and a new magnetic beads made in China,and then typed with Identifiler system in ABI3130XL Genetic Analyzer.210 of these samples were also quantitated by ABI7500 Real Time System.Results Total of 9100 genomic DNA was extracted from various forensic biological samples by the new magnetic beads made in China and automation workstation methods,and most of them were successfully typed for STR analysis.In these biological samples,oral swabs and muscles were of the highest Success rate of STR typing(100%),and the lowest was touched cell samples (50.0%).Conclusion The new magnetic beads made in China with automation workstation methods can be applied to DNA extraction of most forensic biological samples.

Objective To study the correlation with ketamine and its metabolite norketamine concentration in plasma and saliva in acute toxic rabbits.Methods Experimental rabbits were given intragastric(i.g.group,n=6)and intravenous(i.v.group,n=6)administration of ketamine respectively,and control tabbits(n=6)were given a same volume of physiologic saline.The plasma and saliva were collected before and after ketamine administration.Ketamine and norketamine in plasma and saliva were determined using GC/MS and GC.The correlation with ketamine(norketamine)concentration in plasma and saliva were artalyzed by a double variable Pearson correlation analysis.Results The correlation coefficients(r)of ketamine (norketamine)concentrations in plasma and saliva were from 0.80 to 0.95 in ketamine i.g.group and i. v.group.Conclusion There is a good relativity between the concentration of ketamine(norketamine)in plasma and in saliva.The ketamine(norketamine)concentration in saliva can be used to estimate the plasma concentration in the forensic identification of ketamine abuse.

Objective To establish a method of solid phase extraction-reversed-phase high performance liquid chromatography(SPE-HPLC)for determination of methomyl in plasma of rat.Methods The sample pretreatment method,the test conditions,the linear range,the sensitivity,the specificity,the precision, the accuracy,the stability and the recoveries for plasma were investigated by using rat plasma spiked with standard methomyl and intemal standard substance.Results The linear range was 0.1～20μg/mL ( r= 0.9993,P<0.001).The limit of detection was 0.03μg/mL(S/N ≥3).The intra and inter-day precision of assay for methomyl was less than 8.33%and 11.11%in plasma respectively.The intra and inter-day accuracy of assay for methomyl was between 90%and 120%in plasma respectively.The recoveries for methomyl were more than 88%±4.4%in plasma.Conclusion The HPLC method for quantitative and qualitative analysis of methomyl is simple,rapid and accurate,which is suitable for the identification of methomyl in the cases.

Objective To examine the effect of Caffeine on the cultured cortical neuron apoptosis in neonatal rats.Methods The primary cerebral cortex neurons for cultures were obtained from neonatal mice 2-3 days after birth,Caffeine reconstituted at final concentrations 300μmol/L and 1 000μmol/L was added to the cell cultures and continuously co-incubated for 6-36 h,respectively after the cortical neurons were continuously cultivated 7 days after incubation under temperature of 37℃ incubator with 5% CO_2 and 100% relative humidity,the intracellular calcium concentration,mitochondrial membrane potential and apoptosis rate were determined by the flow cytometry.The activity of Caspase-9 was assayed by enzyme-labeled instrument,and Caspase-9 activity by the enzyme-1inked analyzer.Cell morphological changes were observed under electron microscope and fluorescent microscope after being stained with Hoechst 33258 fluorescent dye.Results Compared with the control group,the average increase in intracellular calcium fluorescence intensity was most significant(P<0.05),which elevated from the normal value 43.13±2.02 to 45.28±1.16 and 46.92±1.99,respectively at 6 h;mitochondrial membrane potentials were reduced most significandy(P<0.05).from the base value 443.58 ±11.77 down to 289.53±16.47 and 165.14±14.72,respectively at 8h.Caspase-9 activity was peaked(P<0.05),from the normal value 1.00±0.000 to 5.33±1.02 and 8.33±0.92,respectively at 10 h.The neuronal apoptosis ratio was increased significantly (P<0.05),from the normal value 4.94±1.74 to 15.98±2.03 and 18.70±2.09,at 36h.The apoptotic bodies were observed at 24 h after administration of 300 μmol/L and 1000 μmol/L Caffeine.Conclusion Caffeine may promote neuronal apoptosis in neonate mice.

Objective To observe the expression of S100β protein in the traumatic brain injury and investigate its relation to the severity of the TBI patients.Methods To collect 30 volunteer controls,30 patients with traumatic brain injury and 30 patients with trauma expect traumatic brain injury.according Glasgow Coma Scale(GCS),TBI patients were divided into tow groups,the minor group is GCS≥8,the severegroup is GCS<8.ELISA method was used for observing the expression of S100β protein in serum from the controls and patients.Results Within 6 hours after TBI,the concentration of S100β protein increased higher in patients of TBI than the others(P<0.05).The concentration of S100β protein increased higher in the severe group(GCS<8)than the minor group(P<0.05).The higher level of seium S100β protein,the more severe of TBI patients,the higher level of serum S100β protein.Conclusion The serums S100β protein can be a special index for the early diagnosis of TBI,the higher level of it,The more severe of patients.