Objective To construct a model combining with traditional Chinese medicine(TCM)syndrome for predicting the risk of disease progression in patients with idiopathic membranous nephropathy(IMN)by machine learning methods,thus to quantitatively evaluating the value of TCM syndrome in the prediction of the risk of disease progression in IMN.Methods Monofactor analysis,recursive feature elimination(RFE)and multivariate binary Logistic regression analysis were used to screen the independent related factors affecting the risk of disease progression of IMN,and then a risk prediction model was constructed.A total of 102 patients with IMN were randomly assigned to the training set and the test set in a ratio of 65∶35,and then the comparison was conducted in the performance indicators of accuracy,sensitivity,specificity,F1 value,and area under the receiver operating characteristic(ROC)area under the curve(AUC)of the risk prediction model with or without the inclusion of the TCM syndrome information.Results Before the inclusion of TCM syndrome information,12 clinical characteristic variables for patients with MN were obtained after monofactor analysis combined with RFE screening,and they were age,hemoglobin quantification,urinary occult blood,24-hour urine protein quantification,urine protein-creatinine ratio,estimated glomerular filtration rate(eGFR),creatinine,uric acid,alanine transaminase,anti-phospholipase A2 receptor antibody(PLA2R-Ab),total cholesterol,and low-density lipoprotein cholesterd.A risk cholesterol prediction model containing the above variables was constructed.The multivariate binary Logistic regression analysis showed that the differences of the clinical variables mentioned above between the training-set group and test-set group were statistically significant,and the risk prediction model presented good sensitivity and predictability.Monofactor analysis combined with RFE screening was performed again after the inclusion of TCM syndrome information,and then 14 variables were obtained,which included blood stasis syndrome and dampness obstruction syndrome.The sensitivity and specificity of the model with the inclusion of the TCM syndrome information were significantly improved when compared with those without the inclusion of TCM syndrome information.Conclusion The results of the study initially indicate that TCM syndrome can be used as an important supplementary variable for predicting the risk of disease progression in IMN,and will provide a reference for intelligent diagnosis through the integration of traditional Chinese and western medicine information,and will supply the guidance for the treatment of IMN with TCM.

DNA mixture has been a problem to be conquered for a long time in the forensic study. The DNA mixtures can be mainly divided into two categories: one comes from the sex assaults, for which we have to detect the sperms among the large amount of vaginal epithelial cells; the other one is the combination of different common samples, such as blood mixtures, salvia-blood mixtures and so on. In recent years, deeper and deeper study on DNA mixtures has led a way to objectiveness and pluralism. Novel techniques, like fluorescence- or magnetic-activated cell sorting strategies,micromanipulation and acoustic different analysis can be utilized to separate sperms in the mixtures; as for the second sort, the application of massively parallel sequencing(MPS), microfluidic droplet, whole-genome amplification, emulsion PCR(ePCR) et al. rise up the chances to detect the minor contributor in the mixtures. Furthermore, the development of both traditional and novel biomarkers which includes STR, Y-STR, SNP, DIP, Microhaplotype, DIP-STR, SNP-STR,, mtDNA-SNP and so on, provide us more analyzing choices in mixture study. The last part of the assay focuses on the latest progresses of evaluating the number of contributors, the explanation theory and calculation software.

Objective To observe anomalous genotypes of 4 multi-copy RM Y-STRs in Han population in Hubei province. Methods 252 unrelated male samples were ampliifed using reported and newly designed primers, then detected and analyzed by AB 3130 genetic analyzer. Results A total of 25 anomalous multi-band patterns were observed in 20 samples corresponding to an incident rate of 7.94%. 5 anomalous genotypes were observed in DYF387S1 locus, 15 in DYF399S1, 1 in DYF403S1 and 4 in DYF404S1. Four samples showed extra alleles in more than one locus. Conclusion Anomalous genotype has high incident rates in RM Y-STR markers and requires extensive attention in forensic practice.

In recent years, the cases of prenatal paternity testing gradually increased in forensic practice. The traditional prenatal paternity analysis can be performed only after invasive sampling of chorionic villi or amniotic fluid, which can result in a risk of miscarriage. The existence of circulating cell-free fetal nucleic acid in maternal plasma has brought new opportunities for the noninvasive prenatal paternity testing. In this paper, the research situation and application prospect of circulating cell-free fetal nucleic acid in maternal plasma in prenatal paternity testing are reviewed.

Forensic medical talents with creative ability should have ability to find.analyze and solve problems from complex cases.According to teaching practice on the basis of PBL this article illustrates application methods of PBL teaching characters in the training of forensic medical talents with creative ability.

Three SNaPshot multiplex assays were developed to test 23 coding region single nucleotide polymorphisms (SNPs) and one control region SNP outside hypervariable regions (HVR)I and II, which was aimed at increasing the discrimination power of the mitochondrial DNA (mtDNA) typing in forensic casework, and confirming haplogroup assignments of mtDNA profiles in both human population studies and medical research. The selected SNPs targeted the East Asian phylogeny. These multiplex assays were validated by comparing with the sequencing analysis of samples chosen randomly. The mtDNA variations of 100 unrelated individuals from the Wuhan population in China were examined and classified into 31 haplotypes, and the haplotype diversity was estimated to be 0.952. The multiplex SNaPshot method is rapid and robust, and suitable for large-scale screening studies of mtDNA variability.

Three SNaPshot multiplex assays were developed to test 23 coding region single nucleo-tide polymorphisms (SNPs) and one control region SNP outside hypervariable regions (HVR) Ⅰ and Ⅱ, which was aimed at increasing the discrimination power of the mitochondrial DNA (mtDNA) typing in forensic casework, and confirming haplogroup assignments of mtDNA profiles in both hu-man population studies and medical research. The selected SNPs targeted the East Asian phylogeny. These multiplex assays were validated by comparing with the sequencing analysis of samples chosen randomly. The mtDNA variations of 100 unrelated individuals from the Wuhan population in China were examined and classified into 3 i haplotypes, and the haplotype diversity was estimated to be 0.952. The multiplex SNaPshot method is rapid and robust, and suitable for large-scale screening studies of mtDNA variability.

Objective To develop a simplified bisulfite genomie sequencing(BGS)method for DNA methylation marker scanning.Methods According to modified BGS protocol,the desalt DNA treated with bisulfite were directly used for bisulfite-PCR(BSP)without alkali treatmenL Complement of the bisulfite modification Wag accomplished by a prolonged pre-denaturation stage.After BSP,a second round PCR was performed with a pair of GC tagged primers to adjust the GC content of the amplieon for direct sequencing.To assess this improved protocol,promotor methylation of TNF-α gene in 3T3-L1 cell and androgen receptor(AR)gene in Hela cell was investigated.The real time BSP for Alu was also used to compare the sensitivity of the modified assay with traditional assay.Results Both the hypermethylated TNF-α promotor and hypomethylated AR promotor were successfully sequenced by improved BGS method,and the results were consistent with that of the traditional assay.The conversion rate reached 100%,while the conversion specificity was higher than 93.75%.The sensitivity of improved BGS method inereaged significantly(t=2.978 2,P＜0.05)and showed good reproducibility.Condusion The improved BGS method is simple and sensitive,facilitating more ambitious genomic methyhtion mapping studies.

Objective To study the Application of X-linked differentially methylated polymorphism site in forensic medicine.Methods X-STR HUMARA was chosen as a model locus.PCR procedures were performed after digestion using methylation-sensitive restriction endonucleases.STR polymorphism of HUMARA was analyzed and compared in samples collected from male and female individuals.Result After digestion with methylation-sensitive restriction enzyme HpaⅡ,no PCR products were obtained from male samples,whereas PCR products from the female samples were normally typed.In monoclonal tumor cell samples from females,only one allele was detected.Conclusion The differentially methylated X-STR HUMARA locus is a novel marker for mixture analysis of mixed stains,sex determination and discrimination of tumor tissues.

Objective To develop a PCR-based method of detecting plankton 16S rDNA for the i dentification of death by drowning. Methods Fifteen Sprague-Dawley rats were divided randomly into three groups: the death by drowning group, the group of submerging after death and the control group. After sacrificing by different ways, the brain, liver, kidney and lung of rats were taken out respectively and DNA were extracted from the tissues of these organs and were amplified subsequently by specific primers selected from the third and fourth variable regions of plankton 16S rDNA. Results The specific amplification products were detected from all 5 samples of lung tissue ( 100% ) , 4 samples from liver and kidney tissues (80% ) , and 3 samples from brain tissue (75% ) in the group of death by drowning. No amplification product was detected in all samples of the control group and the amplification product was detected only in 1 sample of lung (20% ) in the group of submerging after death. Conclusion The PCR-based method of detecting plankton 16S rDNA for the identification of death by drowning is certainly feasible.