Forensic Analysis of 43 Medical Disputes Caused by Death after Cardiac Surgery / 法医学杂志

Objective To explore the causes and characteristics of medical disputes caused by death after cardiac surgery and to analyze the pathological changes after cardiac surgery and the key points of forensic anatomy, thus to provide pathological evidence for clinical diagnosis and treatment of cardiac surgery and judicial appraisal as well as reference for the prevention of medical disputes in such cases. Methods Forensic pathological cases of medical disputes caused by death after cardiac surgery which were accepted by the Center for Medicolegal Expertise of Sun Yat-Sen University from 2013 to 2018 were analyzed retrospectively from aspects such as causes of death, pathological diagnosis, surgery condition, medical misconduct, and so on. Results The causes of death after cardiac surgery of 43 patients were abnormal operation, low cardiac output syndrome, postoperative infection, postoperative thrombosis, and other diseases. Among the 43 cases, there were 18 cases without medical fault while 25 cases had medical fault. Conclusion The medical disputes caused by death after cardiac surgery are closely related to the operative technique and postoperative complications. The causes of medical faults include defects in diagnosis and treatment technique, as well as unfulfillment of duty of care.

Strategy of the Causes of Death of Dependents / 法医学杂志

Objective To discuss the methods and strategies to identify the causes of dependents' deaths, as well as provide the experiences that can be used for reference and scientific basis for the forensic identification of the potentially growing deaths of the same kind in the future. Methods The 13 cases concerning death of dependents accepted by Sun Yat-sen University Forensic Center were collected, and the basic information of the dependents were statistically described. The nutritional status, environmental condition and medical care condition were evaluated according to dietary energy, living space, environment and medical treatment condition. Results Among the 13 dependents, there were 11 males and 2 females, with the oldest 74 and the youngest 9 and dwelling time was from 0.4 to 5.6 years. Forensic pathological examination showed that 13 dependents had infectious diseases and 11 were severely dystrophic. There were no fatal mechanical injuries or poisoning in dependents. Molecular pathological screening of 4 cases revealed no pathogenic variants of sudden death susceptible genes. The poor status of the diet, nutrition, living environment and medical care of these dependents were discovered. The direct cause of death of all 13 dependents was identified to be disease. The lack of nutrition, poor living environment and lack of medical care were thought to play a dominant role in causing the deaths of 12 dependants. Conclusion The death identification should follow the judicial procedure. In identification of the causes of death and analysis of the proportion of the affecting factors resulting in death, all factors, including nutrition,environment, medical care, injury and diseases, need to be considered.

Research Progress of the Correlation between Caveolin and Unexpected Sudden Cardiac Death / 法医学杂志

Due to the negative autopsy and without cardiac structural abnormalities, unexpected sudden cardiac death （USCD） is always a tough issue for forensic pathological expertise. USCD may be associated with parts of fatal arrhythmic diseases. These arrhythmic diseases may be caused by disorders of cardiac ion channels or channel-related proteins. Caveolin can combine with multiple myocardial ion channel proteins through its scaffolding regions and plays an important role in maintaining the depolarization and repolarization of cardiac action potential. When the structure and function of caveolin are affected by gene mutations or abnormal protein expression, the functions of the regulated ion channels are correspondingly impaired, which leads to the occurrence of multiple channelopathies, arrhythmia or even sudden cardiac death. It is important to study the effects of caveolin on the functions of ion channels for exploring the mechanisms of malignant arrhythmia and sudden cardiac death.

Correlation between Genetic Variants and Polymorphism of Caveolin and Sudden Unexplained Death / 法医学杂志

OBJECTIVES@#To explore the genetic variation sites of caveolin （CAV） and their correlation with sudden unexplained death （SUD）.@*METHODS@#The blood samples were collected from SUD group （71 cases）, coronary artery disease （CAD） group （62 cases） and control group （60 cases）, respectively. The genome DNA were extracted and sequencing was performed directly by amplifying gene coding region and exon-intron splicing region of CAV1 and CAV3 using PCR. The type of heritable variation of CVA was confirmed and statistical analysis was performed.@*RESULTS@#A total of 4 variation sites that maybe significative were identified in SUD group, and two were newfound which were CAV1： c.45C>T （T15T） and CAV1：c.512G>A （R171H）, and two were SNP loci which were CAV1：c.246C>T （rs35242077） and CAV3：c.99C>T （rs1008642） and had significant difference （P<0.05） in allele and genotype frequencies between SUD and control groups. Forementioned variation sites were not found in CAD group.@*CONCLUSIONS@#The variants of CAV1 and CAV3 may be correlated with a part of SUD group.

Distribution of human enterovirus 71 in brainstem of infants with brain stem encephalitis and infection mechanism / 法医学杂志

OBJECTIVE@#To explore the mechanism that how human enterovirus 71 (EV71) invades the brainstem and how intercellular adhesion molecules-1 (ICAM-1) participates by analyzing the expression and distribution of human EV71, and ICAM-1 in brainstem of infants with brain stem encephalitis.@*METHODS@#Twenty-two brainstem of infants with brain stem encephalitis were collected as the experimental group and 10 brainstems of fatal congenital heart disease were selected as the control group. The sections with perivascular cuffings were selected to observe EV71-VP1 expression by immunohistochemistry method and ICAM-1 expression was detected for the sections with EV71-VP1 positive expression. The staining image analysis and statistics analysis were performed. The experiment and control groups were compared.@*RESULTS@#(1) EV71-VP1 positive cells in the experimental group were mainly astrocytes in brainstem with [dark]-brown particles, and the control group was negative. (2) ICAM-1 positive cells showed [dark]-brown. The expression in inflammatory cells (around blood vessels of brain stem and in glial nodules) and gliocytes increased. The results showed statistical difference comparing with control group (P &lt; 0.05).@*CONCLUSION@#The brainstem encephalitis can be used to diagnose fatal EV71 infection in infants. EV71 can invade the brainstem via hematogenous route. ICAM-1 may play an important role in the pathogenic process.

Application of serum total IgE, tryptase and chymase in the identification of death caused by drug anaphylactic shock / 法医学杂志

OBJECTIVE@#To explore the application value of serum total IgE, tryptase and chymase in the identification of death caused by drug anaphylactic shock.@*METHODS@#The general information from 235 cases of non-drug anaphylactic shock and 32 cases of drug anaphylactic shock were analyzed. The serum IgE level had been detected in the cases. Ten cases caused by coronary disease and 10 cases caused by sudden manhood death syndrome were selected from non-drug anaphylactic shock cases for the control group. Expressions of tryptase and chymase in the lung and heart were detected using immunohistochemistry method. The number and IOD of positive mast cells were counted.@*RESULTS@#In the drug anaphylactic shock group, the IgE value of 18 samples (56.25%) was significantly higher than the normal upper limit of 120 IU/mL. In the non-drug anaphylactic shock group, the IgE value of 67 samples (28.51%) was higher than 120 IU/mL. The expressions of tryptase and chymase were significantly increased in lung and myocardial tissue in drug anaphylactic shock group (P < 0.05).@*CONCLUSION@#Tryptase and chymase are more superior than that of the serum total IgE in the diagnosis of death caused by drug anaphylactic shock, and are more suitable in forensic practice.

Effect of structural domain II of HCV 5' noncoding region on its translation initiation activity / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To examine the role of domain II of hepatitis C virus (HCV) 5' noncoding region (5' NCR) in its translation initiation activity.</p><p><b>METHODS</b>The fragment of HCV 5' NCR with deletions of 5'-proximal 118 nucleotides were amplified with PCR, then was used to substitute for the full length HCV 5' NCR of plasmid pCMVNCRluc, a firefly luciferase (Fluc) eukaryotic expression plasmid regulated by HCV 5' NCR, to generate recombinant plasmid pCNl-d3. pCMVNCRluc, pCNl-d2 (modified pCMVNCRluc with deletions of HCV 5' NCR ntl-43) and pCNl-d3 were transfected into HepG2 cells with liposome transfection protocol, respectively, and the relative luciferase activity was measured to analyze the regulatory effect of the truncated 5'NCR on Flue gene expression. Meanwhile the Flue mRNA levels were detected by RT-PCR.</p><p><b>RESULTS</b>The recombinant plasmid was successfully constructed. The Flue mRNA levels of the 3 plasmids were not significantly different (P > 0.05), and there were also no significant difference between the relative luciferase activity of plasmids pCNl-d2 and pCMVNCRluc (P > 0.05). However, that of pCNl-d3 was decreased significantly (P < 0.01, compared with pCNl-d2 or pCMVNCRluc).</p><p><b>CONCLUSION</b>Structural domain II of HCV 5' NCR plays an important role in its translation initiation activity.</p>

Influence of human cytomegalovirus infection on cell cycle and replication licensing factor Cdt1 in human embryonic lung fibroblastic cells / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the influence of human cytomegalovirus (HCMV) infection on cell cycle and the expression of replication licensing factor Cdt1 in human embryonic lung fibroblastic (HEL) cells and to explore the pathogenesis of HCMV infection.</p><p><b>METHODS</b>HEL cells were synchronized in the G0/G1 phase by the serum starvation method. The synchronized HEL cells were infected with HCMV, and those that were not subjected to HCMV infection were used as the control group. The HEL cells were harvested at 12, 24, 48, 72 and 96 hrs of HCMV infection. The cell cycle of HEL cells was detected by the flow cytometry. The expression of Cdt1 mRNA in HEL cells was determined by reverse transcription-polymerase chain reaction (RT-PCR).</p><p><b>RESULTS</b>The cells in the G1 phase in the control group was significantly more than in the HCMV-infected group 12 and 24 hrs after infection (P < 0.01). The expression of Cdt1 mRNA in the HCMV-infected group was significantly lower 12 and 24 hrs after infection but increased significantly 48 hrs after infection compared with the control group (P < 0.05). The expression of Cdt1 mRNA reached a peak at 12 hrs of infection in the control group, but at 48 hrs of infection in the HCMV-infected group, which markedly lagged behind the control group.</p><p><b>CONCLUSIONS</b>HCMV infection arrests the cell cycle of HEL cells at the G1 phase. HCMV infection makes Cdt1 expression delay. HCMV infection can interfere cell cycle of HEL cells possibly through affecting the expression of Cdt1.</p>

Apoptosis of cultured cortical neurons of rat's brain induced by heroin / 法医学杂志

OBJECTIVE@#To investigate whether heroin can directly induce apoptosis in primary cultured cortical neurons of rat's brain.@*METHODS@#Cultured primary neurons cultures were obtained from cerebral cortex of embryo rats. After 7 days, the cells were incubated with different concentrations of heroin (purity-80%) for 24 hours. The neuronal survival was assessed by cell viability counting with fluorescent diacetate (FDA) staining. The morphological and biochemical changes were observed with Hoechst 33258 fluorescent staining and then analyzed by agarose gel electrophoresis, respectively.@*RESULTS@#After treatment with different concentrations of heroin, the neurons showed a decreased survival rate in a dose dependent manner, and there was a significant difference in the survival rate between the heroin group and the control group (P < 0.05). When exposed to different concentrations of heroin, neurons exhibited the morphological and biochemical features of apoptosis, including cell shrinkage, neurite degeneration, network disappearance, condensation and aggregation of nuclear chromatin, and the formation of DNA ladders. With the increase of heroin concentration of rat's brain more apoptotic bodies were seen.@*CONCLUSION@#Heroin can directly induce apoptosis in primary cultured cortical neurons in rat's brain.

Effects of serum from crush injury rats on vascular endothelial cell apoptosis and their potential mechanism / 法医学杂志

OBJECTIVE@#To investigate the effects of serum from crush injury rats on vascular endothelial cell apoptosis and their potential mechanism.@*METHODS@#Bovine aorta endothelial cells were cultured in vitro and the effects of serum from crush injury rats on cell apoptosis and intracellular calcium concentration ([Ca2+]i) were observed. Meanwhile, the levels of rat blood plasma endothelin-1 (ET-1) and atrial natriuretic peptide(ANP) were measured.@*RESULTS@#Compared with normal rat serum treatment, the cell apoptosis rate decreased from (8.26+/-1.75)% to (2.75+/-0.90)%, while the concentration of [Ca2+]i increased from (96.98+/-3.95) to (118.79+/-3.22) nmol/L in serum from crush injury rats, respectively. The concentration of ET-1 and ANP increased significantly in crush injury rat serum.@*CONCLUSION@#Serum from crush injury rats could inhibit apoptosis of the vascular endothelial cells. These effects may be related to increased level of [Ca2+]i mediated by ET-1 and ANP.

The effects of heroin on intracellular free Ca2+ of rat myocardium / 法医学杂志

OBJECTIVE@#To observe the effects of heroin on intracellular free Ca2+ in rat myocardium.@*METHODS@#The effects of heroin on intracellular free Ca2+ were observed in cultured neonatal rat myocardium by measuring intracellular free Ca2+ concentration using calcium fluorescent probe Flou-3/AM and laser scanning confocal microscope.@*RESULTS@#Different doses and concentrations of heroin appeared to have different effects on intracellular free Ca2+ concentrations, with a dosage dependent short linear increase in the fluorescence intensity (i.e., Ca2+ concentration) leading to [Ca2+]i peak.@*CONCLUSION@#Heroin could affect concentrations of [Ca2+]i in myocardium and its dosage related effect needs further investigation.

Cloning of distinguishing DNA sequences of Gastrodia elata Blume and application of them in identifying gastrodia tuber / 生物工程学报

Gastrodia elata Bl. is a famous and costful traditional Chinese medicine. Their genomic DNA fingerprints were investigated using a modified Randomly Amplified Polymorphic DNA method. DNA fragments common to all or to fine populations were identified and recovered. Five DNA fragments were proven not to be reported through DNA cloning, PCR identifying, nucleotide sequencing and bioinformatics analyses and were received in and recorded by NCBI GenBank. Gastrodine contents of the Gastrodia tuber samples were determined using high performance liquid chromatography technique. The distribution of the five DNA fragments in 9 Gastrodia elata Blue populations and the correlation with gastromedicine content were studied. The results show the distribution of these DNA sequences varied greatly among the populations whereby DNA Sequence 1 was the common and distinguishing molecular marker for all the populations studied and DNA Sequence 2 may relate to higher gastrodine content. In conclusion, these DNA marker sequences can be employed to identify genuine gastrodia tubers, better varieties and optimize their selection and cultivating.

Cellular mechanism of heart injury in the early stage of crush injury in rats / 法医学杂志

OBJECTIVE@#To study cellular mechanism of cardiomyocytes injury in the early stage of crush injury by observing some effects of crush injury rat sera on cultured neonatal rat cardiomyocytes.@*METHODS@#One to three days old neonatal rat cardiomyocytes were cultured in vitro and some effects of crush injury rat sera on beating rate, cell surface area, total protein content, 3H-Leu incorporation, intracellular calcium concentration ([Ca2+]i) and Fos protein expression were observed in cultured rat cardiomyocytes.@*RESULTS@#Compared with normal rat serum group, crush injury rat sera decreased beating rate(beats/min) of cardiomyocytes from 88.3 to 26.4, cell surface area, total protein content, 3H-Leu incorporation, [Ca2+]i (nmol/L) and PI of Fos protein expression were increased.@*CONCLUSION@#Crush injury rat sera suppress cell beating, increase intracellular calcium, induce Fos protein synthesis and cause cell hypertrophy, which may cause cardiac injury in the early stage of rush injury.

Establishment and the significance of a cell model of secreted alkaline phosphatase co-controlled by HCV 5'NCR and NS3 serine protease / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To establish a cell model of secreted alkaline phosphatase (SEAP) co-controlled by HCV 5'NCR and NS3 serine protease in an effort to develop new antiviral agents.</p><p><b>METHODS</b>The fragments of HCV 5'NCR and NS3/4A-SEAP were amplified by PCR. They were fused into pBluescript SK+ to generate 5'NCR-NS3/4A-SEAP chimeric plasmid. The resulting chimeric gene was subcloned into HindIII/Bsu36 I site of pSEAP2-Control (a SEAP eukaryotic expression plasmid), to generate pNCR-NS3/4A-SEAP, in which the SEAP was fused in-frame to the downstream of NS4A/4B cleavage site. The SEAP activity in the culture media of transiently transfected cells was monitored quantitatively. The regulatory effect of HCV 5'NCR and NS3 serine protease on SEAP expression was measured by treatment of transfected cells with antisense oligodeoxynucleotide (ASODN) against HCV 5'NCR and TPCK, a irreversible serine protease inhibitor.</p><p><b>RESULTS</b>The SEAP activity in the culture media reached 80801+/-4794 RLU, and was significantly inhibited by 5 micromol/L, 10 micromol/L of ASODN (t=4.315, p<0.01; t=6.985, p<0.001) and 100 micromol/L of TPCK (t=6.949, P<0.001).</p><p><b>CONCLUSION</b>A cell model of SEAP co-controlled by HCV 5'NCR and NS3 serine protease has been successfully established. This might promote the screening of anti-viral drugs</p>

Improvement of Methods of Making Myocardial Ischemia-Reperfusion Model and Effects on Electrocardiogram and Ultrastructure of Myocardium in Rats / 实用儿科临床杂志

Objective To improve the methods of making myocardial ischemia-reperfusion model and observe the changes of electrocardiogram and ultrastructure of myocardium in rats.Methods The chest of young male SD rats through the fourth intercostal space was opened,and the left coronary artery was tied with a silicagel tube,after 30 minutes,untied to perfuse.Changes of electrocardiogram were observed and recorded.After reperfusion,the levels of AST,LDH,and CK-MB were measured and the tissue samples of the infarct areas were examined by transmission electronic microscope.Results The 90 percent of total rats were made myocardial ischemia reperfusion model successfully.In myocardial ischemia reperfusion rats:the QRS wave of myocardial ischemia-reperfusion rats was much higher than that of control group;the level of cardiac enzymes increased;myocardial and vascular endothelial cells ultrastructure was damaged seriously.Conclusions The improvement of modus operandi is right.Ischemia reperfusion can cause evident damage of myocardial and vascular endothelial cells ultrastructure in rats,and damage of myocardial cells is more severe than vascular endothelial cells.

The changes of electrocardiogram and serum cardiac troponin I at the early stage of crush injury in rats / 法医学杂志

OBJECTIVE@#To observe the changes of electrocardiogram and serum cardiac troponin I at the early stage of severe crush injury in rats.@*METHODS@#Crush injury was produced in Sprague-Dawley rats. The changes of electrocardiogram were recorded with the standard II, the serum levels of cardiac troponin I were studied by automated chemiluminescence assay.@*RESULTS@#The ST segment elevated considerably after crush injury and lasted 24 h, the levels of serum cTnI were much higher than those of the control groupes after 6 h of injury.@*CONCLUSION@#Cardiomyocyte injury was induced in the early phase of crush injury.

The preliminary study on change of serum enzyme level in crush syndrome rat / 法医学杂志

OBJECTIVE@#To study the changes of serum creatinine kinase(CK) and its cardiac-specific isoenzyme compound(CK-MB) levels in crush injury rats.@*METHODS@#Crush injury was produced in SD rats, the serum levels of CK and CK-MB were studied by automated biochemical analyzer.@*RESULTS@#The levels of plasma CK and CK-MB were much higher in crush injury rats than those of the control group.@*CONCLUSIONS@#Cardiomyocyte injury may be induced in the early stage of crush injury rats.

The changes of tryptase activity in anaphylactic shock guinea-pigs / 法医学杂志

OBJECTIVE@#To investigate the changes of the activity of tryptase of sera, lungs and bronchial tubes in the guinea-pigs which suffered from hetero-serum anaphylactic shock.@*METHODS@#Sera and tissues were collected from anaphylactic shock guinea-pigs, and the enzyme activity was tested colormetrically using special substrate, BAPNA.@*RESULTS@#The activity of tryptase of sera, lungs and bronchial tubes increased significantly in Anaphylactic guinea-pigs compared with control group.@*CONCLUSION@#The changes of tryptase activity are helpful to diagnose anaphylactic shock.