Objective Corruption is the most common cadaver phenomenon in forensic practice and an important basis for inferring time of death(PMI),but the definition of corruption degree and the construction of model inference models have always been difficult in the practice of forensic science.Methods In this study,the late postmortem phenomena were observed.Meanwhile,the microbial flora structure of gut and gravesoil and the nature of gravesoil were detected,for analyzing the changes before and after the key moment of abdominal rupture which naturally happened during the cadaver decay.Results The results found that from the macroscopic and microscopic levels,there were significant differences in cadaver decay,including microbial flora structure and gravesoil properties before and after the key moment of the natural abdominal rupture during cadaver decay.The phenomena are highly observable and can be accurately judged by forensic examinations,as well as related means in the field of biology and physiochemistry.In this study,this critical event was called Rupture Point.Conclusion The Rupture Point can be used as an important node for the assessment of cadaver decay degree in the practice of forensic medicine.It can be utilized for a cut-off point as well when constructing PMI inference models based on microbial flora structure changes.The accuracy of PMI inference models can be improved when the models were constructed in segments.

Objective To explore the feasibility of tracing the source of human biological samples by microbiome,and provide new ideas for case investigation.Methods Biological samples from buccal mucosa,foot arch,armpit and other parts of 10 volunteers were collected for three consecutive months.The microbial community structure of the samples was confirmed by using the 16SrRNA gene sequence information.And the microbial community diversity analysis and random forest classification prediction model were carried out.Results There were significant differences in the microbial community structure of the three parts of human body.In this study,a prediction model of random forest classification with an accuracy of more than 90%was successfully constructed.Conclusion In this study,a classification and prediction model based on the microbiome information of human biological samples was constructed to judge the source location,which broadened the forensic application scenarios of human microorganisms.

The DNA characterization of Cannabis sativa L.has been one of the key directions of anti-drug research at home and abroad.Previous research mainly focused on the identification of cannabis-species and gender differentiation,and have constructed a number of corresponding composite amplification systems.With the rapid development of high-throughput sequencing technology,the whole genome of C.sativa and the sequences of key enzyme genes for its major physicochemical components have been sequenced successively,and intra-species differentiation studies of C.sativa based on specific molecular markers have gradually emerged.However,due to the high variability of cannabis subspecies-and variety-specific molecular markers,relevant foreign studies failed to provide ideal molecular marker support for the identification of intra-specific distinctions of Cannabis sativa in China.Based on this,this paper comprehensively analyzes the current situation and shortcomings of domestic and international research on intra-specific differentiation of C.sativa,and combines the previous research results of this group to elaborate on how to use high-throughput sequencing technology to solve the problem of the lack of intra-specific molecular markers of C.sativa in China.

Purpose: Bone destruction and pain caused by cancer is one of the most devastating complications of cancer patients with bone metastases, and it seriously affects the quality of patients’ life. Extracellular matrix metalloproteinase inducer (EMMPRIN) is a cell adhesion molecule with increased expression in a variety of tumors. This study focused to clarify the specific function of EMMPRIN in bone metastasis of breast cancer. Materials and Methods: Adenovirus with shRNA-EMMPRIN was transfected into MRMT-1 rat breast carcinoma cells, and the MRMT-1 cells with different expression levels of EMMPRIN were implanted into the bone marrow cavity of rat tibia. Next, the effect of down-regulation of EMMPRIN was evaluated as follows: bone damage was detected by X-ray radiological and tartrate-resistant acid phosphatase staining; the tumor burden was evaluated by hematoxylin and eosin staining; the test of pain-related behaviors was assessed used the bilateral paw withdrawal mechanical threshold; and the levels of secretory factors in tumor conditioned medium were determined by using enzyme-linked immunosorbent assay. Results: We found that down-regulation of EMMPRIN in tumor cells can simultaneously reduce tumor burden, relieve cancer-induced bone destruction and pain. Conclusion: Materials and Methods EMMPRIN is expected to be a therapeutic target for relieving bone metastasis of breast cancer and alleviating cancerinduced bone destruction and pain. The method of targeting EMMPRIN may be a promising strategy for the treatment of cancer in the future.

Purpose: Bone destruction and pain caused by cancer is one of the most devastating complications of cancer patients with bone metastases, and it seriously affects the quality of patients’ life. Extracellular matrix metalloproteinase inducer (EMMPRIN) is a cell adhesion molecule with increased expression in a variety of tumors. This study focused to clarify the specific function of EMMPRIN in bone metastasis of breast cancer. Materials and Methods: Adenovirus with shRNA-EMMPRIN was transfected into MRMT-1 rat breast carcinoma cells, and the MRMT-1 cells with different expression levels of EMMPRIN were implanted into the bone marrow cavity of rat tibia. Next, the effect of down-regulation of EMMPRIN was evaluated as follows: bone damage was detected by X-ray radiological and tartrate-resistant acid phosphatase staining; the tumor burden was evaluated by hematoxylin and eosin staining; the test of pain-related behaviors was assessed used the bilateral paw withdrawal mechanical threshold; and the levels of secretory factors in tumor conditioned medium were determined by using enzyme-linked immunosorbent assay. Results: We found that down-regulation of EMMPRIN in tumor cells can simultaneously reduce tumor burden, relieve cancer-induced bone destruction and pain. Conclusion: Materials and Methods EMMPRIN is expected to be a therapeutic target for relieving bone metastasis of breast cancer and alleviating cancerinduced bone destruction and pain. The method of targeting EMMPRIN may be a promising strategy for the treatment of cancer in the future.

Ethics review as the content of the research management of forensic medicine,is the research object of forensic medicine.Focusing on ethics review of the forensic medicine research,this paper discussed on the necessity of ethics review and guiding principles of ethics committee,and appealed for the construction of ethics review in forensic medicine research.

Objective To prepare the standard molecular weight fragment mixtures. Methods Primers were designed to prepare clones which contained different sizes of standard molecular weight fragments. The template used for amplification of insert fragments was the pMD18-T vector. Bacteria culture and plasmid extraction were used to obtain abundant target fragment. Unlabeled DNA fragments were prepared by double digestion of the recombinant plasmids, and the fluorescent adaptor was prepared by annealing with two partial reverse complimentary DNA fragments. The unlabeled fragments and fluorescent adaptor were connected by DNA ligation reaction assisted with T4 DNA ligase. In this way, different sizes of standard molecular weight fragments were prepared. Standard molecular weight fragment mixture was finally prepared by mixing all the fragments together before purification. Results Ten standard molecular weight fragments of different sizes were prepared. The sizes of each fragment are 80bp, 124bp, 194bp, 224bp, 254bp, 304bp, 349bp, 399bp, 424bp and 454bp. The internal standard could accurately determine the size of PCR products amplified with the DNATyper15 kit. Conclusion Using this method, the standard molecular weight fragment mixture which meet the requirements of research and laboratory use was prepared, perfectly providing a new method for preparation of the DNA molecular weight standards. The peaks and the size of the prepared DNA internal lane standard are correct, which can be used to calculate the DNA fragments size in capillary electrophoresis.

Objective To evaluate the effect of hyperoxygenated solution on myocardial injury in the rats with acute carbon monoxide (CO) poisoning.Methods Thirty pathogen-free adult male SpragueDawley rats,weighing 250-300 g,were randomly divided into 5 groups (n=6 each) using a random number table:control group (C group),acute CO poisoning group (ACP group),and different doses of hyperoxygenated solution groups (HP1-3 groups).CO 120 ml/kg was injected intraperitoneally to establish the model of acute CO poisoning.Hyperoxygenated solution 10,15 and 20 ml/kg were infused via the caudal vein at 1 h after intraperitoneal injection of CO in HP1-3 groups,respectively.At 24 h after intraperitoneal injection of CO,blood samples were collected from the caudal vein for determination of plasma creatine kinase (CK),creatine kinase-MB (CK-MB),lactic dehydrogenase (LDH) and alpha-hydroxybutyrate acid dehydrogenase (α-HBDH) activities using the automatic biochemical analyzer.The rats were then sacrificed,and myocardial specimens were obtained for examination of the pathological changes with a light microscope.Results Compared with group C,the plasma LDH,α-HBDH,CK and CK-MB activities were significantly increased in ACP and HP1-3 groups (P＜0.01).Compared with group ACP,the plasma LDH,α-HBDH,CK and CK-MB activities were significantly decreased in HP1-3 groups (P＜0.05 or 0.01).Compared with group HP1,the plasma LDH,α-HBDH,CK and CK-MB activities were significantly decreased in HP2,3 groups (P＜0.05).The pathological changes of myocardium were significantly attenuated in HP1-3 groups as compared with group ACP.Conclusion Hyperoxygenated solution can attenuate myocardial injury in the rats with acute CO poisoning.

Objective To construct a rapid genetic DNA extraction method, with nano magnetic beads, self-designed reagents system and extracting process. Method Part I: DNA extraction from old blood cotton swab sample with self-designed DNA extraction kit, then quantiifed by UV spectrophotometer. The method was further optimized on the preliminary results. Part II: All kinds of difficult DNA sample were tested with optimized kit, to detect the applicability of the kit. Result By improving the experimental condition, the extraction effects of different DNA sample is good, meanwhile, the extraction cost is relatively low.

Transcriptional regulation is one of the major regulations in plant adventious shoot regeneration, but the exact mechanism remains unclear. In our study, the RNA-seq technology based on the IlluminaHiSeq 2000 sequencing platform was used to identify differentially expressed transcription factor (TF) encoding genes during callus formation stage and adventious shoot regeneration stage between wild type and adventious shoot formation defective mutant be1-3 and during the transition from dedifferentiation to redifferentiation stage in wildtype WS. Results show that 155 TFs were differentially expressed between be1-3 mutant and wild type during callus formation, of which 97 genes were up-regulated, and 58 genes were down-regulated; and that 68 genes were differentially expressed during redifferentiation stage, with 40 genes up-regulated and 28 genes down-regulated; whereas at the transition stage from dedifferentiation to redifferention in WS wild type explants, a total of 231 differentially expressed TF genes were identified, including 160 up-regualted genes and 71 down-regulated genes. Among these TF genes, the adventious shoot related transcription factor 1 (ART1) gene encoding a MYB-related (v-myb avian myeloblastosis viral oncogene homolog) TF, was up-regulated 3 217 folds, and was the highest up-regulated gene during be1-3 callus formation. Over expression of the ART1 gene caused defects in callus formation and shoot regeneration and inhibited seedling growth, indicating that the ART1 gene is a negative regulator of callus formation and shoot regeneration. This work not only enriches our knowledge about the transcriptional regulation mechanism of adventious shoot regeneration, but also provides valuable information on candidate TF genes associated with adventious shoot regeneration for future research.
Arabidopsis ; growth & development ; Arabidopsis Proteins ; physiology ; Gene Expression Regulation, Plant ; Genes, Plant ; Plant Shoots ; growth & development ; RNA ; Regeneration ; Seedlings ; growth & development ; Transcription Factors ; physiology ; Up-Regulation