Objective To observe the effect of treating generalized anxiety disorder with clearing heart and relieving rstlessness acupuncture combined with western medicine. Methods 87 patients with generalized anxiety disorder were randomly recruited into a control group and a treatment group. Both groups received conventional western medical treatment. On this basis, the control group was given conventional acupuncture therapy, while the treatment was given clearing heart and relieving restlessness acupuncture. The efficacies were evaluated with Hamailton anxiety (HAMA)scale and clinical global impression scale (CGI). The adverse reaction was evaluated with treatment emergent symptoms scale (TESS). Results One week after treatment, the efficacies and adverse reaction in the treatment group markedly decreased comparing with those before the treatment(P<0.05). Two weeks after treatment, the efficacies and adverse reaction of both groups decreased comparing with those before the treatment(P<0.01). Besides, the decrease continues with time passing by. One week after treatment, the HAMA score in the control group markedly decreased comparing with that of the control group (P<0.05), and the CGI score in the treatment group had no significant difference comparing with that of the control group(P<0.05). Conclusion The treatment of clearing heart and relieving restlessness acupuncture on generalized anxiety disorder has rapid effect and good compliance.

Objective To genotype mixed samples with next generation sequencing and evaluate its prospects in forensic DNA application. Methods Three mixed biological samples from rapes cases and their reference samples were collected. DNA was extracted using the MagAttract M48 DNA Manual Kit(200). The ForenSeqTMDNA Signature Prep Kit was used for library preparation, and next generation sequencing was performed on the MiSeq FGx system. The ForenSeqTMUniversal Analysis v1.2.1 software was used for data analysis. NGS-based STR results were compared with CE-based genotypes. Results A single length polymorphic STR allele in the mixed profile could be recognized as two sequence polymorphic STR alleles from different donors, which would assist mixed profile analysis. Such phenomenon was observed in D3S1358, D9S1122 and D13S317 in this work. Conclusion Our results suggested that precision STR genotyping of mixed samples based on NGS can provide more information and hints for mixed STR profile separation.

Capillary electrophoresis-based STR genotyping is accepted as the gold standard for human individual identification. Next generation sequencing (NGS) allows for the full resolution of STR base composition, and has the potential to be widely used in the field of forensics. Compared with length polymorphism, STR sequencing could provide more information, and quantitatively calculating the forensic parameters is necessary. In this paper, we established simple models for length-based and sequence-based STRs, and calculated the forensic parameters for both models. The results showed that for a single STR locus, compared with length polymorphism, STR sequence polymorphism could provide higher power of discrimination and power of exclusion, indicating sequence-based STR marker have stronger ability for identifying unrelated individuals and exclude non biological father. By combining 15 non-linkage loci for forensic DNA analysis, the cumulative matching probability values for length-based and sequence-based STR models are at 10-18and 10-26levels, respectively. Only 10 non-linkage sequence-based STR is required to reach a cumulative matching probability of as high as 15 length-based STR loci. It is hoped that these simulated models and calculations can provide a reference for the forensic application of NGS-based STR genotyping.

Objective Sequencing depth is a key parameter in next generation sequencing,which is closely related to the accuracy of sequencing results.Forensic biological evidence examination requires extremely high accuracy.It is crucial to scientifically and reasonably set the sequencing depth analysis threshold for forensic next generation sequencing testing.Methods This study used targeted sequencing data of microhaplotypes from 50 samples with known genotypes.By calculating the accuracy,precision,recall,and F1 score of each locus under various threshold conditions,two types of analysis threshold setting methods,which were based on fixed read count and fixed sequencing depth ratio,were studied extensively.Results The results showed that false positives were observed when the analysis threshold was set at 50×or 100×.When the analysis threshold was set at 200×,false negatives were observed.When the analysis threshold was set at 1.5%,3.0%,or 4.5%,false positives were observed.This study further proposed a third type of analysis threshold setting method,which was based on sequencing depth ratio scatter plots.With this method,no false positive or false negative was observed in the results.This article then explored four factors that lead to significant differences in the sequencing depth of forensic next generation sequencing experiments,compared with the analysis threshold setting method for capillary electrophoresis technology,and discussed the correlation between analysis thresholds and the ability to distinguish mixed DNA.Conclusion Employing the sequencing depth ratio scatter plot method to set analysis threshold has significant application value in next generation sequencing-based forensic genetic marker genotyping.