Trichostatin C attenuates TNFα-induced inflammation in endothelial cells by up-regulating Krüppel-like factor 2 / 药学学报

Krüppel-like transcription factor 2 (KLF2) plays a key regulatory role in endothelial inflammation, thrombosis, angiogenesis and macrophage inflammation and polarization, and up-regulation of KLF2 expression has the potential to prevent and treatment atherosclerosis. In this study, trichostatin C (TSC) was obtained from the secondary metabolites of rice fermentation of Streptomyces sp. CPCC 203909 as a KLF2 up-regulator by using a high throughput screening model based on a KLF2 promoter luciferase reporter assay. TSC significantly inhibited the adhesion of tumor necrosis factor-α (TNFα) induced monocytes (THP-1) to human umbilical vein endothelial cells (HUVECs). Western blot results showed that TSC decreased TNFα induced the protein expression increase of vascular cell adhesion molecule-1 (VCAM-1), and thereby inhibited endothelial inflammation. The results of histone deacetylase (HDAC) overexpression and molecular docking experiments showed that TSC upregulated the expression of KLF2 by inhibiting subtypes of HDAC 4/5/7. In conclusion, this study suggests that TSC up-regulates the expression of KLF2 through inhibiting HDAC 4/5/7 and thus inhibits TNFα induced endothelial inflammation, and it has the potential to prevent and treat atherosclerosis.

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.

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.

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.

Relationship between DNA degradation and postmortem interval of corrupt corpse / 法医学杂志

OBJECTIVE@#To study the relationship between DNA degradation and postmortem interval of corrupt corpse.@*METHODS@#By determining the marrow DNA content with histochemical technique and image analysis.@*RESULTS@#The content of marrow DNA decreased gradually with prolongation of postmortem interval, and it evencould be detected till 14 days after death.@*CONCLUSION@#There was a linear relationship between the degradation rate of the nuclear DNA and postmortem interval.

A study on the relationship between the degradation of protein and the postmortem interval / 法医学杂志

OBJECTIVE@#To observe the degradation of actin and tubulin in the liver tissue of rats after death and to find an objective indicator of the postmortem interval (PMI).@*METHODS@#Female rats were killed under anesthesia by ether and incubated at 21 degrees C in a temperature controlled system to simulate postmortem changes for 18 days postmortem. Protein in the hepatic tissue was extracted, actin and tubulin were then examined by western blot. Thereafter, the semi-quantitative analysis of the image of western blot was performed.@*RESULTS@#Actin in the liver tissue of rats could be detected at 8 days postmortem, but could not be examined after 10 days postmortem. beta-tubulin rather than alpha-tubulin could be examined after 2 days postmortem, and beta-tubulin could not be examined at 4 days postmortem.@*CONCLUSION@#There is some difference in the degradation between actin and tubulin, their different preservation period postmortem may be used as a parameter for PMI estimation.

A preliminary study on estimation of postmortem interval according to beta-actin mRNA stability in rat / 法医学杂志

OBJECTIVE@#In order to find a new parameter to estimate the postmortem interval, beta-actin mRNA in lung and thoracic muscle of rats was detected at different time point postmortem.@*METHODS@#Rats were killed by neck dislocation and left in a temperature controlling system at 21 degrees C for 12 days postmortem. Total RNA in lung and thoracic muscle at different time point was extracted and beta-actin mRNA expression was examined by RT-PCR. Semi-quantification analysis of the image of electrophoresis was performed to confirm the changes of beta-actin mRNA expression.@*RESULTS@#beta-actin mRNA in lung of rats still could be detected at 12 days postmortem, but-disappeared in thoracic muscle at 8 days postmortem.@*CONCLUSION@#The expression of beta-actin mRNA in lung and thoracic muscle could be a new parameter for estimation of PMI.

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.

Fed-batch fermentation of Escherichia coli that express fab fragment of anti-HBsAg / 生物工程学报

To develop a fed-batch fementation process of E. coli TOP10 containing a recombinant plasmid pBAD/HBs Fab. Cells were grown in semi-defined medium at 37 degrees C, and the feed operation using glycerol as carbon source was performed when dissolved oxygen increased. When the target cell concentration reached to 64g/L, arabinose was added to a final concentration of 0.02%. Cells were grown for another 5h with the culture temperature decreased from 37 degrees C to 30 degrees C. In the whole process, cell growth was monitored by measuring OD600 of samples taken at 1/2h intervals and the dissolved oxygen was kept above 30%. After the fementation, E. coli pellets were collected for purification of Fab protein. The specificity of Fab protein was confirmed by Western blot, and binding activity to HBsAg was verified by Dot blot. Cell concentration we got is 96g wet bacteria per liter, the Fab protein is about 6% of total protein of the host, that is 80mg per liter. Stable fermentation parameters were obtained for fermentation to improve productivity of the Fab protein. The Fab protein was produced in the form of soluble biologically active protein, it's better than inclusion bodies from which biologically active protein can only be recovered by complicated and costly denaturation and refolding process.

Interpretation of DNA profiles for mixed stains / 法医学杂志

This article review the advance in interpretation of mixed forensic stains using DNA profiling, including autosome STR profiling, sex profiling determined by PCR, Y-specific STR profiling, mitochondrial DNA profiling and single nucleotide polymorphism profiling. The statistics methods for mixed stain has also been reviewed.

The relationship between postmortem degradation of marrow DNA in bosom bone and late postmortem interval estimation / 法医学杂志

OBJECTIVE@#To study the relationship between degradation of marrow DNA and late postmortem interval (PMI).@*METHODS@#Marrow were left on natural condition for 0,1,3,5,7 day after death respectively, Marrow DNA were detected by using Feulgen staining and computerized image analysis.@*RESULTS@#The content of marrow DNA could be detected till 7 days after death yet.@*CONCLUSION@#The degradation of marrow DNA may be used on estimation the late PMI.

A preliminary study of epidemiology of sudden manhood death syndrome in the peasant derived workers in Dongguan city / 法医学杂志

OBJECTIVE@#To study the feature of sudden manhood death syndrome(SMDS) in Dongguan city.@*METHODS@#The data of 284 cases of SMDS were analysized by retrospective study.@*RESULTS@#The distribution of age, sex, the time of death and hometown of the dead in SMDS were described. The clinical or anatomical characters of SMDS were also discussed.@*CONCLUSION@#These data will contribute to the late epidemical study.

The unusual typing of short tandem repeats / 法医学杂志

Short tandem repeats (STRs) have been widely used in forensic sciences such as stain analysis and paternity testing. Although most of STR typing could give the reliable and clear results, some unusual typing have been observed in forensic practice. The anomalous typing could result from a lot of causes, including DNA genetic variation, poor quality or quantity of DNA template, different typing system or method, nonspecific reaction in PCR or anomalous electrophoresis migration. The unusual results may disturb the right interpretation of STR typing.

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.

A quantitative analysis of peptidergic innervation in sinoatrial node in cases of sudden manhood death syndrome / 法医学杂志

OBJECTIVE@#To study the distribution and proportion of neuropeptide containing nervers in the sinus node in cases of sudden manhood death syndrome (SMDS) and to explore the mechanism of SMDS.@*METHODS@#Immunohistochemical staining and quantitative analysis of neuropeptide Y (NPY) and vasoactive intestinal peptide(VIP) in the sinus node in 6 cases of SMDS and in 12 cases of non-cardiac death(control group) were achieved by LSAB method and computerized image system.@*RESULTS@#As for NPY positive materials, VIP positive materials and the ratio of VIP/NPY in the sinus nodes, there were no significant difference between the control group and SMDS group.@*CONCLUSION@#The mechanism of SMDS and the abnormality of autonomic nervous innervation in the sinoatrial nodes maybe incorrelation.

Injury by electrical forces: pathophysiology, biophysics and pathogenesis / 法医学杂志

As the technologic sophistication of generation and distribution of electrical energy has grown, so has the general concern about the effects of electric fields on human health. There can be no doubt that the significance of electrical trauma will continue to grow with our increasing use of power. It is apparent that our understanding of the various forms of electric trauma must increase, while we continue to promote safety near electrical hazards and develop effective medical therapies. Tissue damage as a result of electrical injury occurs by two mechanisms which are summative in action and have a variable degree of contribute to the ultimate damage produced. Thermal tissue damage occurs as a result of heat generated within the tissue (which offer an electrical resistance) secondary to the passage of the electrical current. High temperatures can also lead to cell membrane components, e.g., phospholipids, to dissolve. Electroportation damage is the tissue damage induced secondary to the strong electric field. Transmembrane potentials caused by electrical current result in the formation of pore in the phospholipid component of the cell membrane resulting in loss of function of the cell membrane with consequent cell death.

Immunohistochemical study with depletion of cardiac troponin T in rats on early myocardial ischaemia / 法医学杂志

OBJECTIVE@#To study the depletion of cardiac troponin T in Rats of early myocardial ischaemia.@*METHODS@#Immunohistochemical method (LSAB) was used and the depletion area of CTnT in myocardial ischaemia were measured by the computer image analysis.@*RESULTS@#There is obviously depletion of CTnT after 15 min in myocardial ischaemia of rats, the mean of CTnT depletion of myocardial ischaemic group is significant difference compared with that of the control group (P < 0.01), and the depletion area increased with the prolongagtion of ischaemic time.@*CONCLUSION@#The depletion of CTnT is one of sensitive signs of early myocardial ischaemia.

Study on the specificity of complement C5 for the postmortem diagnosis of myocardial infarction / 法医学杂志

In order to explore the specificity of complement C5 in the postmortem diagnosis of myocardial infarction, changes of C5 staining in normal, infarcted and other non-infarcted myocardia with direct or indirect myocardial injuries (myocarditis, mechanical asphyxia, electrocution, hemorrhagic shock, cardiac contusion and organophosphate poisoning) were studied with immunohistochemistry and image analysis. The results showed that positive C5 staining could be observed in groups of myocardial infarction and myocarditis, but not in groups of mechanical asphyxia, electrocution, hemorrhagic shock, cardiac contusion, and organophosphate poisoning. It is indicated that positive reaction of C5 could only be affected by myocarditis, which means that it was more specific for the diagnosis of myocardial infarction.

Histopathological observation of experimental slight viral myocarditis / 法医学杂志

To study the diagnostic method of slight viral myocarditis in the field of forensic pathology, slight viral myocarditis model was induced in Balb/c murine by coxsackie virus B3. Organs of hearts, livers, spleens, lungs and kidneys were examined through routine pathological methods. Pathological changes at different levels of these organs were observed. The results indicated that viral myocarditis was a kind of disease with multiple organ alterations and that the pathological observation and comprehensive analysis of multiple organs was one of the useful methods for diagnosing slight viral myocarditis.

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.