With the rapid development of Next-generation Sequencing(NGS)technologies, and its high throughput and low cost is applied widely in the field of life science, the increase in the depth of sequencing together decrease in the consumption of time and cost, makes a wide application of NGS in the research of microbiological research, ancient DNA study, clinical diagnosis, forensic science research, etc. The article discusses the second generation sequencing technology platform and its genetic markers in the forensic application. Included STR typing, SNP typing, HLA genotype prediction and the application in the degradation of the material.

Objective To explore species specificity of 29 Y-SNP loci and to lay the foundation for forensic application.Methods Human DNA and 8 different kinds of common animals' DNA were amplified separately by PCR.The PCR products were analyzed with PAGE.Results Twenty-three out of 29 Y-SNP loci were only amplified by the male human DNA,which indicated a good species specificity.Three loci amplified from human and some animals showed that the length of the product was different between two species.Another three loci amplified from human and animal showed that the length of the fragment was similar between human and animal.Five loci were amplified by PCR products from both male and female samples.Conclusion Most of 29 Y-SNP loci,amplified in male humans,have good species specificity and can be directly used for personal identification and paternity testing.

Objective To evaluate the efficiency of combined screening for chromosomal abnormalities in the first trimester and the ultrasound characteristics of these fetuses.Methods Retrospective study for 5000 singleton pregnancies by combined screening of trisomies 21,18,13 and Turner syndrome.Risk algorithms were developed for calculation of patient-specific risks for each of the three trisomies based on maternal age,fetal nuchal translucency,free β human chorionic gonadotropin and serum pregnancy associated plasma protein A at 11 to 13 +6 weeks of pregnant.The value of nuchal translucency (NT) and β-hCG and pregnancy-associated plasma protein A (PAPP-A) level were inputted computer,and calculate the risk value (≥ 1 ∶ 270) by automatic analysis software.Two hundred and four cases with high risk were performed transabdominal chorionic villus biopsy to detect the fetal chromosomal karyotypes.Meanwhile,other ultrasonic characteristics of fetal were elevated.Results (1) Five thousand cases of pregnant women were detected,including 4983 normal cases,62 cases were induced labor for a variety of reasons in the second trimester,including 40 cases with normal karyotype but with congenital heart disease,17 cases of chromosome abnormalities (9 cases trisomy 21,2 cases trisomy 18,1 cases trisomy 13,4 cases 45X),2 cases spina bifida,2 cases digestive tract obstruction,1 cases giant bladder.One case with low risk of fetal chromosomal abnormalities in combined screening,but high risk of age (maternal age were over 40 years old),it was 21 trisomy syndrome after the prenatal diagnosis.(2) Five cases of nasal bone loss in 9 cases of trisomy 21 (5/9),5 cases with three tricuspid regurgitation (5/9),4 cases of venous ductus a wave flow reverse (4/9),3 cases of fetal nasal bone loss accompanied by tricuspid regurgitation and venous ductus a wave flow reverse (3/9).One case of nasal bone loss in 2 cases of trisomy 18,2 cases were tricuspid regurgitation and venous ductus a wave flow reverse.Two cases in 4 cases of 45X had venous ductus a wave flow reverse.There were 8 cases (0.16％) nasal bone absence in 4983 cases of normal karyotype fetus,48 cases (0.96％) of tricuspid regurgitation and 44 cases (0.88％) of venous ductus a wave flow reverse.Thirty-two cases in 40 cases (80％) of fetal congenital heart disease were tricuspid regurgitation,30 cases of venous ductus a wave flow reverse (75％).Eight cases of nasal bone absence normal karyotype fetus were found the nasal bone at 20 weeks gestation.Conclusion Combination screening of nuchal translucency with serum markers in the first trimester were high detection rate and low false positive rate; a wave reversion and fetal nasal bone absence accompanied by tricuspid regurgitation can improve the detection rate of abnormal karyotype; abnormalities ultrasound marker may be associated with fetal congenital heart disease at 11-13 +6 weeks of pregnancy.

Objective To study the D gene polymorphism of Rh-negative blood group in Kunming resident population with PCR technology for gene typing,and to establish and perfect PCR detecting method for RhD gene.Method 46 samples of Rh-negative blood samples were collected and PCR amplification with exon-SSP had been done firstly.The samples of D gene part existing were screened,then PCR amplifying by intron-SSP had been conducted.The PCR amplification production with intron-SSP was used to DNA sequence.Results Among 46 Rh-negative samples,the number of D gene lost intacfly was 29;the complete existing one with all exons was 12.The sequencing results were that 2 samples of only containing D10 were RhD-CE(3-9)-D and the sample with only D4 and D6 was the newly discovered variant allele RhD-CE(2-3,5,7-9)-D.RhD ? was not found in our study.Conclusion(1)There is high frequency of RhD-CE(3-9)-D in Kunming area.(2)A new variant allele of RhD-CE(2-3,5,7-9)-D is first found in Kunming area.

The reform for the laboratory operational mechanism includes laboratory management system reform and experimental teaching reform.The author made some explorations and research to the operational mechanism and management pattern of university laboratory of forensic biology,then presented some ideas.

To introduce tutorial system for undergraduate in the forensic biology teaching is an important measure of educational reform of forensic medicine.The article presented some schemes and experiences about the practice of tutorial system for undergraduate in the forensic biology department of Kunming Medical College.

Objective The study aimed to investigate 15 autosomal loci polymorphism(such as D3S1358 and TPOX)in Nujiang Bai nationality,in order to construct genetic basic data of Bai population and provide a foundation for population study and forensic cases. Methods Venous blood was collected from 124 unrelated Bai individuals in Nujiang,Yunnan province. DNA was extracted by classical organic solvent extraction. PCR technique was applied to amplify the sequence of autosomal loci. The products were measured by electrical fluoroscopy. The gene frequencies of 15 STR loci were investigated and genetic polymorphisms were analyzed. ResultsGene polymorphisms of 15 STR loci were detected and the distributions of genotype conformed with Hardy-Weinberg balance law(P > 0.05). The cumulative match probability(CPM)was 4.869×10-17,the cumulative excluding probability(CPE)was 99.999 99%,and the combined power of discrimination(CDP)was 99.999 97%. ConclusionThe 15 STR loci used in this study were highly polymorphic informative content in Bai population and could be applied to population study and forensic practice.

Objective To explore the effect of liver-soothing therapy on the expression of estrogen receptors mRNA in perimenopausal syndrome (PMS) rats with liver qi stagnation. Methods A total of 30 nature aging rats are assigned into control groups (n=8), model groups (n=8),Chaihu-Shugan San (CHSGS group,n=8) andDanzhi-Xiaoyao San (DZXYS group, n=8), according to the random number table. The PMS liver-Qi stagnation syndrome rat models were established by the methods of isolation raised and chronic bondage in all the groups except the control group. CHSGS group were administered 4.0 g/kg water decoctions ofChaihu ShuganSan, and DZXYS group 4.9 g/kg water decoctions ofDanzhi XiaoyaoSan respectively for 3 weeks after the rat models established. The model group and control group were administered with equal volume of normal saline. The open field test was used for the behavior test. The serum E2, FSH, LH level were measured by radioimmunoassay. The ERα, ERβ in ovary were measured by quantitative real-time PCR. Results Compared with model group on the 21st days, the CHSGS and DZXYS groups showed a significantly increase in crossings (49.6 ± 6.0, 51.6 ± 5.8vs. 40.0 ± 4.6,P<0.05 orP<0.01) and rearings (14.1 ± 0.7, 14.6 ± 2.3vs. 10.9 ± 1.8,P<0.05 orP<0.01). Cmpared with the model group, the FSH (3.96 ± 0.48 mIU/mlvs.5.31 ± 0.41 mIU/ml) significantly decreased in the CHSGS group, and the LH (6.65 ± 0.46 mIU/mlvs. 8.10 ± 0.62 mIU/ml) significantly decreased in the DZXYS group (P<0.05). Compared with the model group, ER alpha mRNA expression (7.42 ± 2.54, 4.91 ± 1.76vs. 3.80 ± 1.36,P<0.01) significantly increased in the CHSGS group, and the ER beta mRNA expression (3.56 ± 0.95vs. 3.10 ± 1.12,P>0.05) increaed in the DZXYS group, but there was no remarkable difference. Conclusion The Liver-soothing therapy can improve the behavior of PMS rats with liver-Qi stagnation, and the mechanism may be related to the regulation of endocrine and ovarian estrogen receptors.

Objective To investigate the molecular mechanism of protection of ischemia preconditioning on renal ischemia reperfusion injury. Methods Male C57/BL6N mice were randomly divided into two groups: in IR group, 35 min ischemia was induced by occlusion of both renal pedicles followed by 24 h perfusion (I/R). 15 min ischemia was induced 4 days before I/R in IPC group. Blood sample and kidney were collected in IR and IPC group after 24 h perfusion. Serum creatinine (Scr) and histological changes were used to evaluate the renal injury. PHD2 and HIF-1αwere evaluated by Western blotting, miR-21 expression was confirmed by real-time PCR. In vitro, hypoxic model was established by 1% O2 in HK-2 cells. Knockdown of miR-21 in hypoxic model was perfermed by locked nucleic acid modified-anti-miR-21 transfection. The levels of miR-21, HIF-1α and PHD2 mRNA were confirmed by real-time PCR. The levels of HIF-1α and PHD2 proteins were tested by Western blotting. Results In vivo, Compared with IR group, the renal function and histological changes were improved in IPC group (P<0.01). Compared with IR group, the expression of miR-21(P<0.01) and HIF-1α(P<0.05) were increased in IPC group, while PHD2 was reduced (P<0.01). In vitro, hypoxia reduced miR-21. The inhibition of miR-21 could increased the expression of PHD2 (P<0.05). Conclusions Ischemia preconditioning may exert protection against renal ischemia reperfusion injury by inhibiting PHD2.

Forensic biology is a main subject of forensic medicine.With the rapid development of science and technology,the courses should be constantly improved.New teaching mode has come into being application of multimedia technology in theoretical lessons,case teaching,independent completion of experimental lessons,which is thanks to the constant reinforcement of teaching staff and the reform of course content and teaching methods.