Prokaryotic expression and immunogenicity analysis of latent infection antigen Rv2628c-Rv1737c fusion protein of Mycobacterium tuberculosis / 中国人兽共患病学报

According to the codon characteristics of the prokaryotic system,we synthesized the optimized Rv2628c-Rv1737c nucleic acid sequence and constructed the expression plasmid pET24a-Rv2628c-Rv1737c.After sequencing,expression in E.coli was induced.The fusion protein was found in inclusion bodies.After renaturation and affinity chromatography purifi-cation,the Rv2628c-Rv1737c recombinant protein,with a molecular weight of 57 kDa and a purity exceeding 90％,was ob-tained.The recombinant protein was used to stimulate PBMC cells from different patients,and the differences in IFN-γ mRNA expression were analyzed with q-PCR.The Rv2628c-Rv1737c fusion protein stimulated patients with TB,particularly latent TB infection(LTBI).The level of IFN-γ mRNA in PBMC cells was higher than that in healthy controls(P＜0.05).Mice immu-nized with BCG+Rv2628c-Rv1737c/DMT showed significant induction of high levels of IgG antibodies.Rv2628c-Rv1737c re-combinant protein,as a latent infection antigen,is recognized by PBMCs cells infected with TB,with strong immunogenicity,and thus may serve as a potential TB subunit vaccine target antigen.This protein may be used to prevent TB infection,particu-larly latent infection,and to perform laboratory diagnosis.

Application of ITO Method in Determination of Special Half-Sibling Relationship / 法医学杂志

OBJECTIVES@#To identify whether the relationship between Zhang A, Zhang B, Zhang C and Zhang X is the half-sibling relationship whose mother is sister (hereinafter referred to as the special half-sibling relationship) or the common first cousin relationship and discuss the application of ITO method in discriminating the special kinship.@*METHODS@#DNA was extracted from blood stain of four identified individuals, PowerPlex® 21 System and AGCU 21+1 STR kit were used to detect autosomal STR genetic markers. Investigator® Argus X-12 QS kit was used to detect the X chromosome STR genetic markers, the special half-sibling index (SHSI) and first cousin index (FCI) and their likelihood ratio (LR) were calculated by ITO method.@*RESULTS@#The LR results of SHSI to FCI, which were calculated based on autosomal STR genotyping and the analysis of X-STR genotyping results suggested that the relationship between Zhang A, Zhang B, Zhang C and Zhang X was inclined to be a special half-sibling relationship.@*CONCLUSIONS@#For the identification of special kinship, it is necessary to comprehensively apply various genetic markers according to the case. After the conclusion that shared alleles cannot be excluded from the analysis, ITO method can be further used to establish discriminant assumptions according to the specific case to obtain objective and reliable identification opinions.

Research Progress of Metabolomics in Forensic Pathology / 法医学杂志

Metabolomics is an interdisciplinary subject that rose in the post-genomic era, which focuses on quantitative study of the response of living organisms to outside stimulation and pathophysiological changes, as well as multiple dynamic response of the level of in vivo metabolites caused by genetic mutation. It is extensively used in basic research of system biology, materia medica, clinical medicine, etc. In the forensic field, metabolomics mainly focuses on forensic toxicology, but with the generalization of certain techniques, it's foreseeable that metabolomics has a broad research prospect in forensic pathology. This article summarizes the major analysis techniques and methods of metabolomics, describes the research status of metabolomic techniques in the field of forensic pathology application research, including postmortem interval and death cause. Moreover, this article summarizes and discusses the potential applicable areas, in order to provide reference for relative research and application.

Difference Analysis Based on 16S rRNA Sequencing of Different Soil Bacterial Communities / 法医学杂志

Objective To study the structure and differences of bacterial communities in different soils, and to explore the effectiveness of 16S rRNA sequencing in identification of different soil. Methods Soil samples from 7 places in Shanghai were collected, then bacterial genomic DNA were extracted from them. The fragments of hypervariable region from 16S rRNA sequences were sequenced with high-throughput sequencing techniques. The results were quantified or visualized with bioinformatics software. The differences in diversity and abundance among the three kinds of bacterial communities in soil samples from grassland, forests and beaches were compared and analyzed. Results The statistical differences that existed among the alpha diversity indexes of bacterial communities in soil samples of grassland, forests and beaches had statistical significance. The relative abundance and diversity of bacterial communities in these three kinds of soil were significantly different. Grassland soil had higher Acidobacteria abundance, forest soil had higher Proteobacteria abundance, beach soil had higher Actinobacteria abundance. However, the differences in soil bacterial communities in artificial grasslands, natural grasslands and industrial district grasslands did not have statistical significance. Conclusion 16S rRNA sequencing can effectively distinguish different soils. This method may be able to provide clues for first crime scene inference in criminal cases.

Research Progress on Age Estimation Based on DNA Methylation / 法医学杂志

Age estimation is of great significance in the fields of criminal investigation and forensic identification. It can provide the age information of individuals to judicial departments to facilitate the development of judicial work. In recent years, age estimation methods expanded from the morphological level to the molecular biology level. With the rapid development of epigenetics represented by DNA methylation, and the advancement of DNA methylation detection technology together with the detection platform, many age estimation methods based on DNA methylation biomarkers, or using several biological fluids, such as blood, blood stains, saliva, semen stains, etc. are developed. Currently, researches related to age estimation based on DNA methylation are relatively widely carried out. This paper summarizes the researches on age estimation based on DNA methylation, in order to provide references for related studies and forensic applications.

Research Progress of CircRNA and Its Application Prospect in Forensic Medicine / 法医学杂志

Circular RNA(circRNA)is a type of noncoding RNA with tissue specificity and high stabil-ity, which forms a closed continuous loop and is abundantly expressed in tissue cells. According to re-cent research, the regulatory function of circRNA elucidating in the occurrence and development of dis-ease shows a potential for diagnosing clinical disease and revealing disease mechanism. This paper re-views the biological characteristics, analysis methods of circRNA and its research progress in clinical ap-plication as biomarker, and outlooks its application in the field of forensic medicine.

Brief discussion on the proof standard of traditional evidence of forensic pathology in murder cases / 复旦学报（医学版）

Objective In the legal proceedings of murder cases,we often encounter the problem of proof standard in traditional forensic pathology in deep research,which is lack of systemic discussion in domestic academic field.This paper aimed to discuss this problem.Methods We compared the relevant characteristics of different legal systems on the standard of proof system,analyzed the elements of the forensic pathology evidence related to murder cases,and discussed the problem with the author's judicial practice experience.Results We believe that the cause of death,death time,injury and injury tools are the three most important forensic pathology evidences in the murder of the standard system as the "three pillars".Conclusions Division of standards in traditional forensic pathology evidence in the murder is of great significance to raise the sense of evidence of forensic medicine,and if it exists major flaws in the evidence of forensic pathology,the evidence can be corrected or the proceedings can be terminated at the very beginning of the litigation phase,which is of great value to the savings of judicial resources.

Application of the Peak Area Ratio of STR Loci to Amelogenin Locus in the Estimation of DNA Degradation / 法医学杂志

OBJECTIVE@#To explore the change rules of peak area ratio of STR loci to Amelogenin (AMEL) locus (STR/AMEL), a sex-determining gene in DNA degradation, and to evaluate the application of STR/AMEL value in the estimation of DNA degradation degree.@*METHODS@#DNA was extracted from iliopsoas, and the variations of STR/AMEL value (Penta E/AMEL, Penta D/AMEL, FGA/AMEL) were analyzed after the artificial degradation was made by DNase I, and the changes of these three ratios of the iliopsoas naturally degraded in an outdoor environment were also analyzed. The regression curves were analyzed using the periods of DNA degradation and outside the body as the independent variable (x) and the STR/AMEL value as the dependent variable (y) and three curve equations under two conditions were established.@*RESULTS@#Both under the conditions of artificial and natural degradation, STR/AMEL value had a negative relationship with the degradation time. The relationship between STR/AMEL and degradation time can be well simulated by the cubic function. R2 was over 0.99 under controlled degradation condition and over 0.86 under natural degradation condition.@*CONCLUSION@#The STR/AMEL value (Penta E/AMEL, Penta D/AMEL, FGA/AMEL) is negatively related with the DNA degradation degree, which follows mathematical regression models strictly, and it might be applied to evaluate the DNA degradation degree.

Research Progress of circRNA and Its Significance in Forensic Science / 法医学杂志

RNA has received more attention in the field of forensic medicine and the development of the new biological markers based on RNA shows great significance in the analysis of complex cases. circular RNA (circRNA) is a kind of non-coding RNA which is widely reported recently. Although the regulatory mechanisms of generation and expression are not fully clear, the existing research indicates that circRNA has important biological functions. CircRNA has a cell-type-specific expression with great stability and a high expression level, which makes it meaningful in forensic applications potentially. In this paper, the research progress, the generation and regulation of circRNA as well as its biological characteristics and functions are summarized, which will provide references for related studies and forensic applications.

Study Progress of Mechanisms and Treatment of Neonatal Birth Femoral Fracture / 实用儿科临床杂志

This article is based on the searching from the United States Library of Congress Web Search science 2005 for the literature on the birth femoral fractures,reviewing its incidence,pathogenesis and treatment of such fractures of the latest developments,for the purpose of the provision of orthopedic and obstetric physician reference.Literature showed that neonatal birth femoral fractures might occur whether deli-veries were through the birth canal or by cesarean section.Whenever faced with fetal asphyxia,intracranial hemorrhage,such as critical state,it is critical to save the life of fetal babies,regardless of treatment.The prognosis of neonatal femoral shaft fractures is normally good without deformity left.