Research Progress and Forensic Application on the Pathogenesis of Coronary Artery Spasm / 法医学杂志

Coronary artery spasm(CAS)is a hyper-contraction of segmental coronary artery in response to multiple stimuli. At present, it's still in lack of specific diagnostic indicators of sudden cardiac death caused by CAS. This review summarizes current researches on the mechanisms of CAS and describes the roles of vascular endothelial dysfunction and vascular smooth muscle hypersensitivity in the course of CAS. Furthermore, the molecular mechanisms of the endogenous NO and endothelin-1 cause vascular endothelial dysfunction, and the phosphorylation of MLC2, Rho kinase and endoplasmic reticulum stress related to vascular smooth muscle hypersensitivity are discussed. Meanwhile, the possibility of forensic application for the related molecules on the diagnosis of sudden cardiac death caused by CAS are also explored.

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.

A forensic analysis of alcohol-positive fatal cases in Shanghai during 2007 to 2016 / 复旦学报（医学版）

Objective To characterize alcohol-related deaths which were not caused by traffic accident in Shanghai and to provide data support for alcohol-related policy and intervention.Methods All alcohol-positive fatal deaths collected in 2007-2016 from Shanghai Public Security Bureau were classified based on blood alcohol concentration (BAC) and distributed by year,gender,age,season,cause of death (COD),manner of death (MOD) and combined drugs.Results There was no significant correlation of alcohol BAC with year or season.Males (88.4％) were more than females (11.6％).Age of 20-39 years (57.1％),mechanical injury (70.1％),homicide (64.6％) and combined illicit use (37.2％) were the largest proportions in respective categories of age,COD,MOD and drug combination.With the BAC rising,mechanical injury and homicide showed decreased proportions,while burn and accidents showed increased proportions.Conclusions Male and the age of 20-39 years are the major populations for alcohol-related deaths in Shanghai.With the rise of BAC,mechanical injury and homicide decrease,while burn and accident increase.Among the combined use of alcohol and drugs,death from acute intoxication should be paid special attention.

Expression of EIIIA+ fibronectin in incised wound of rat's skin / 法医学杂志

OBJECTIVE@#To explore the relationship between the expression of EIIIA+ fibronectin in incised wound of rat's skin and injury time.@*METHODS@#The wounding model was established by cutting the dorsal skin of 48 adult SD rats. The rats were sacrificed at the pre-set injury time as immediately, 0.5 h, 1 h, 2 h, 3 h, 4 h, 6 h, and 8 h. The skin samples were taken at the margin of wound. The expression of the EIIIA? fibronectin was detected by immunohistochemistry and Western blotting and the relationship be- tween its expression and injury time was observed. Results The expression of EIIIA+ fibronectin was not observed immediately. The basal cell of skin began to show positive expression 0.5 h after injury. With the extension of injury time, positive staining became stronger. The value of relative optical density was gradually increased with prolonged injury time by the Western blotting analysis.@*CONCLUSION@#The expression of EIIIA+ fibronectin could be used for estimation of injury time in the early stage of skin injury.

Relationship between PMI and relative expression of myocardial various RNAs in rats died of different causes / 法医学杂志

OBJECTIVE@#To observe the changes of relative expression of myocardial various RNAs in rats died of different causes and their relationship with PMI.@*METHODS@#The rat models were established in which the rats were sacrificed by broken neck, asphyxia, and hemorrhagic shock. Total RNAs were extracted from myocardium. The quantitative real time PCR was used to calculate threshold cycle values of RNAs including glyceraldehyde-3-phosphate dehydrogenase (GAPDH), beta-actin, inducible nitric oxide synthase (iNOS), hypoxia-inducible factor-1 (HIF-1), tumor necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6), and U6 small nuclear RNA (U6 snRNA) and to study the changes of the relative expressions of various indexes with PMI.@*RESULTS@#U6 snRNA with stable expression level could be used as appropriate internal control. In the early PMI, the relative expression of GAPDH, HIF-1, iNOS, TNF-alpha, and IL-6 more characteristically increased in groups of asphyxia and hemorrhagic shock than in group of broken neck, but the quantity of beta-actin decreased in all groups. In the late PMI, all the relative expressions significantly declined in correlation with the degradation of RNA.@*CONCLUSION@#The characteristic changes of each RNA expression can be used as references to estimate PMI in deaths by different causes.

Estimation of early postmortem interval using beta-actin mRNA in rat's brain, heart and kidney / 法医学杂志

OBJECTIVE@#To explore the relationship between beta-actin mRNA degradation in SD rat's brain, heart and kidney and early postmortem interval (PMI) in order to find new markers for estimating early PMI.@*METHODS@#Rats were sacrificed and kept in the place at a temperature of 20 degrees C. The total RNA were extracted from the brain, heart and kidney at different PMI points. Real time RT-PCR was applied to determine beta-actin mRNA levels in total RNA and the results were given in the form of Ct values. Linear relationships between PMI and Ct values were obtained and the functions of linear regression were established.@*RESULTS@#The great decrease of beta-actin mRNA level were observed in the three organs. The degradation rate was obviously higher in 24 hours after death in the heart and kidney. However, there were no significant changes in the brain. The changes of Ct values and PMI showed a good linear relationship.@*CONCLUSION@#beta-actin mRNA in rat's brain, heart and kidney degrades obviously after death and can be used for estimating early PMI by its degradation rules.

Characteristics and mechanism of boat propeller injuries / 法医学杂志

OBJECTIVE@#To summarize the characteristics and investigate the mechanisms of boat propeller injuries so as to explore the identification methods between boat propeller injuries and corpse dismemberment.@*METHODS@#More than 100 autopsy cases of boat propeller injuries were collected in a period between 1994 and 2005 in Huzhou district, Zhejiang province. The characteristics of injuries caused by propeller, including abrasion, wound, fracture and severed wound, and the characteristics of clothing, were retrospectively studied and summarized. The severed cross wound section of boat propeller injuries was compared with that caused by corpse dismemberment.@*RESULTS@#The boat propeller injuries were resulted from high-speed propellers with enormous splitting power and mechanical cutting, while corpse dismemberment were resulted from cutting and dismembering the body with sharp instruments. Due to the different mechanisms, the different strength of force and recoil force, the severed wound cross section had different characteristics.@*CONCLUSION@#Wounds caused by boat propeller injuries have their unique characteristics, distinguished from wounds of dismembered corpse.

Pro-apoptotic effect of decorin on rat mesangial cells in vitro / 中华病理学杂志

<p><b>OBJECTIVE</b>To investigate the effects of over-expression of decorin (DCN) gene on apoptosis of cultured rat mesangial cells (MsC).</p><p><b>METHODS</b>PcDNA3.1A-DCN plasmid was transfected into cultured rat MsC by the induction of liposome and positive clones were selected by treating the cells with G418. The MsC clones stably expressing DCN (MsC/DCN) were confirmed by cellular immunofluorescence, reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blot. The DCN-siRNA was used for blocking DCN expression in MsC/DCN, and was confirmed by Western blot. The apoptosis of MsC was assayed by flow cytometry and Hoechst staining. Expression of Caspase-3 was assayed by Western blot.</p><p><b>RESULTS</b>Positive clones with DCN over-expression were established. The apoptotic rate in MsC/DCN was (20.40 +/- 8.01)% and was much higher than the (2.07 +/- 0.99)% in MsC (P < 0.01). Some of the MsC/DCN cells showed typical morphologic changes of apoptosis. The protein expression of active Caspase-3 was also significantly increased in MsC/DCN compared to MsC (P < 0.01). DCN-siRNA transfection not only significantly blocked the expression of DCN and reduced the rate of apoptotic cells, but also down-regulated the expression of active Caspase-3.</p><p><b>CONCLUSIONS</b>Over-expression of DCN induces apoptosis of cultured rat MsC in vitro. This effect of DCN inducing apoptosis suggests a novel strategy for regulating the proliferation of MsC in glomerular diseases.</p>

Anatomy study of MGA in Chinese and its effect on legal expertise / 法医学杂志

OBJECTIVE@#This study aimed to clarify the morphology of the Martin-Gruber anastomosis (MGA) in Chinese.@*METHODS@#One hundred and five Chinese upper limbs (36 males and 20 femalese) were dissected to find the connections between medial nerve and ulnar nerve. The MGA was classified as previously described by Lee.@*RESULTS@#MGA was found in 24 cases (22.9%), in 11 of the 36 male and 5 of the 20 female. There was no obvious difference in the frequency of MGA in both upper limbs. Most MGA ulnar position was located at the medial and distal segment of the forearm.@*CONCLUSION@#MGA anatomy could play important role in forensic diagnosis of ulnar nerve injury in Chinese population.

Expression of EDA-fibronectin in injured human skin fibroblast and its potential application in estimation of wound healing / 法医学杂志

OBJECTIVE@#[corrected] To study the expression of EDA-FN variant in injured human skin fibroblast and its relation to wound healing.@*METHODS@#Human skin fibroblast cells were separated from human skin and cultured and monoclonal antibody to fibronectin [IST-9] was applied to amplify target segments.@*RESULTS@#Trace amount of EDA-FN was detected in the culture cells. The expression was increased within 8 hours after the injury.@*CONCLUSIONS@#The expression of EDA-FN would be an ideal marker for estimation of wound healing

Effect of interferon-gamma on transforming growth factor beta/Smad signal pathway and expression of matrix metalloproteinase-2 and tissue inhibitors of matrix metalloproteinase-2 in cultured rat mesangial cells / 中华病理学杂志

<p><b>OBJECTIVE</b>To study the effect of interferon-gamma (IFN-gamma) on the proliferation of mesangial cells (MsC) and transforming growth factor (TGF)-beta/Smad signal pathway, the mRNA and protein expression of matrix metalloproteinase-2 (MMP-2) and tissue inhibitors of matrix metalloproteinase-2 (TIMP-2), and to provide an experimental basis for IFN-gamma treatment of renal fibrosis.</p><p><b>METHODS</b>Cultured MsC were treated with IFN-gamma at different concentrations and the proliferation of MsC was examined by MTT. Protein and RNA samples were extracted from MsC at 0, 0.5, 1, 2, 4, 6, 12, 24 h after treated by 100 IU/ml IFN-gamma. The mRNA and protein expression of Smad3, Smad7, MMP-2 and TIMP-2 were analyzed by real-time RT-PCR and Western blot, respectively.</p><p><b>RESULTS</b>The expression of Smad7 mRNA and protein were promptly elevated at 0.5 hour after the IFN-gamma treatment and lasted for 6 hours, but the proliferation of MsC was not altered. The elevated expression of Smad3, MMP2 mRNA and proteins persisted after 6 hours, whereas the expression of TIMP-2 mRNA and protein decreased.</p><p><b>CONCLUSION</b>The therapeutic effect of IFN-gamma of renal fibrosis may be mediated by TGF-beta/smads signal pathway through up-regulation of MMP-2 expression, coupled with down-regulation of TIMP-2 expression.</p>

The relationship between expression of FN in human skin injury and early injury time / 法医学杂志

OBJECTIVE@#In order to supply an effective reference of early injury time estimation and explore the time limit of detection of EDA\EDB mRNA in human skin samples, the expression of alternative splicing segment of fibronectin--EDA\EDB in incised wound of human skin were studied.@*METHODS@#Using in situ hybridization with DIG-labeled anti-sense RNA probe, the expression of FN EDA\EDB domain was detected in human skin incised wound at the early stage of injury (from 30 min to 3 h).@*RESULTS@#The positive expression rates of FN-EDA\EDB immediately after injury and area far away from wound were same as the control group. The expression of FN-EDA\EDB in human skin incised wound showed a gradually increased tendency in early injury time (within 3 h). The positive expression cells were mainly distributed in basement cells of epidermis and the expression of EDA is much higher than EDB. It's difficult to detect EDA\EDB mRNA when the samples were deposited in air for 4 hour.@*CONCLUSION@#FN-EDA\EDB may be used as a sensitive mark for the estimation of early injury time. The in-situ hybridization technique is not applicable in the application.

Molecular cloning of recombinant fibronectin EDA and EDB fusion protein / 法医学杂志

OBJECTIVE@#Construct a recombinant plasmid pET28a-EDA-EDB, prepare the fusion EDA-EDB protein.@*METHODS@#For the production of recombinant fibronectin EDA-EDB in Escherichia coli, the EDA and EDB segments were separated from pGEM2-EDA/EDB and recomposed with two additional amino acids, then cloned into the expression vector pET28a. pET system to express EDA-EDB fusion protein and 6 x His/Ni-NTA system to purify it in a single step were used. Western blotting confirmed the purified protein.@*RESULTS@#The EDA and EDB segments were ligated and inserted into pET28a vector. EDA-EDB fusion protein was highly expressed in Escherichia coli BL21 (DE3). Afterwards, it was purified by Ni-NTA resin and verified by western blotting.@*CONCLUSION@#EDA-EDB fusion protein can be expressed in pET system and purified by 6 x His/Ni-NTA system.