Though Runx2 and Osterix are both key transcription factors in the pathway of osteoblast differentiation, whether Runx2 positively regulates Osterix being unknown. It was showed that Runx2 induced the gene expression of Osterix both in the non-osteoblastic cell lines, either pluripotent or differentiated, and in the osteoblastic cell lines. At the same time, the results also indicated that Runx2 up-regulated the activity of the 3.2 kb human Osterix promoter. Further experiments identified a highly conserved and functional Runx2 binding site "AGTGGTT" within the promoter. Thus the results support the hypothesis that Runx2 is involved in the regulation of the Osterix gene expression. Moreover, the transient transfection and dual-luciferase assay showed Osterix up-regulated the activity of the 2.3 kb type Ⅰ collagen promoter in the non-osteoblastic cells, but Runx2 did not. This difference implies that Osterix, the down stream transcription factor of Runx2 during osteoblast differentiation, is needed to stimulate the osteoblast-specific gene expression of type Ⅰ collagen.

Objective T o observe the expression changes of hypoxia inducible factor-1α (H IF-1α) and vascular endothelial grow th factor-A (V E G F-A ) in rats w ith arrhythm ias, and to explore the differences of the expression pattern in the tw o indicators of acute m yocardial ischem ia caused by arrhythm ias and coronary insufficiency. Methods T he arrhythm ia w as induced by C aC l2, and the expression changes of H IF-1α and V E G F-A w ere detected by im m unohistochem istry, W estern blotting and real-tim e PC R w ithin 6 h after the arrhythm ia in rats. Results T he expression of H IF-1α and V E G F-A show ed diffuse in the m yocardial tissue of rats died from arrhythm ias. B oth of them increased in the early arrhythm ia, then decreased. E xtensive m yocardial ischem ia happened at the beginning of arrhythm ia occurrence and its range didn't expand w ith tim e. Conclusion T he expressions of H IF-1α and V E G F-A in m yocardium of the rats w ith arrhythm ia can provide evidence for the differential diagnosis of acute m yocardial is-chem ia caused by fatal arrhythm ia and coronary insufficiency.

Fatal anaphylactic shock is common in forensic practice. However, it is difficult to diagnose for lacking specific pathological and morphologic changes in forensic autopsy. The application of some biochemical indicators is of great significance. This paper reviews the biological characteristics of some biochemical indicators and detection methods. The forensic application, problems and prospects of these indicators are also introduced in details. The stable biochemical indicators, IgE, tryptase and chymase, show great potential and advantages in the identification of fatal anaphylactic shock in forensic medicine.

Objective To explore the changes of serum IgE and tryptase caused by anaphylactic shock rats and discuss the relation to PMI and preservative environm ent of corpse and specim en. Methods Rats were used for establishing anaphylactic shock m odels and random ly divided into room tem perature group, refrigeration group, frozen group, manual hem olysis group, specim en preservation group. And the control group was also established. The blood sam ples were collected after rats were sacrificed. The de-gree of hem olysis was graded according to the color of the upper layer of the serum . The mass concen-tration of IgE and tryptase in each group was detected by ELISA. Results The levels of serum IgE and tryptase in anaphylactic shock dead rats were higher than that of the control group. Room tem perature and frozen m ade obviously differences on the levels of serum IgE and tryptase with various PMI. The levels of serum IgE and tryptase in refrigeration group show ed relatively stable. The levels of serum tryptase and IgE were elevated with differently increasing hem olysis. The levels of serum IgE and tryptase show ed no obvious changes during the specim en kept under different tem perature conditions for 25 days. Conclusion Serum IgE and tryptase obviously increased in anaphylactic shock rats. H ow ever, the levels were influenced by PMI and environm ental tem perature, especially under the conditions of room tem perature and frozen.

The initiation of cellular antiviral signaling depends on host pattern-recognition receptors (PRRs)-mediated recognition of viral nucleic acids that are known as classical pathogen-associated molecular patterns (PAMPs). PRRs recruit adaptor proteins and kinases to activate transcription factors and epigenetic modifiers to regulate transcription of hundreds of genes, the products of which collaborate to elicit antiviral responses. In addition, PRRs-triggered signaling induces activation of various inflammasomes which leads to the release of IL-1β and inflammation. Recent studies have demonstrated that PRRs-triggered signaling is critically regulated by ubiquitin and ubiquitin-like molecules. In this review, we first summarize an updated understanding of cellular antiviral signaling and virus-induced activation of inflammasome and then focus on the regulation of key components by ubiquitin and ubiquitin-like molecules.
Epigenomics ; Immunity, Innate ; Inflammasomes ; Inflammation ; Nucleic Acids ; Pathogen-Associated Molecular Pattern Molecules ; Phosphotransferases ; Porcine Reproductive and Respiratory Syndrome ; Signal Transduction ; Transcription Factors ; Ubiquitin

OBJECTIVETo investigate the clinical characteristics of a Chinese pedigree with cardiac conductive disease complicated by atrial fibrillation and the therapeutic effect of the treatments.

METHODSAll the family members including the proband were screened with routine examination, electrocardiography, echocardiograpy, Holter recording, chest X-ray, blood biochemistry tests and autoantibody test. The proband received dual chamber pacemaker implantation combined with oral amiodarone treatment for 3 months. The patient was monitored for thyreoid function and chest X-ray during the treatments, and was followed up for another 3 months.

RESULTSClinical evidence of organic heart disease was found in none of the family members. The proband showed recurrent dizziness and chest distress, which exacerbated after exercise, and ECG showed atrial fibrillation and severe A-V block. The proband's uncle was found to have atrial fibrillation and III degree A-V block after a syncope episode at the age of 30, and received a pacemaker treatment. Her grandpa died from a heart attack without detailed clinical documentations. No other family members showed abnormal ECG or a history of any heart events. The proband's condition was improved by treatments, after which ECG and Holter recording showed pace rhythm without atrial fibrillation.

CONCLUSIONCardiac conductive disease with atrial fibrillation can present in one family, and can be managed effectively and safely with implantation of dual chamber pacemaker combined with oral amiodarone.

Asian Continental Ancestry Group ; Atrial Fibrillation ; complications ; genetics ; therapy ; Atrioventricular Block ; complications ; genetics ; therapy ; Cardiac Resynchronization Therapy ; Female ; Heart Conduction System ; abnormalities ; Humans ; Pedigree ; Young Adult

In order to regulate the apoptosis induced by Fas-FasL system, a soluble isoform of mouse Fas was cloned from thymocytes of immature mice with the primers designed according to the full-length Fas cDNA sequence in the GeneBank. It was directionally inserted into the intermedium vector pUC19. DNA sequencing proved that it was consistent with the expected sequence. Then it was subcloned into the eukaryotic expression vector pCA13, which was used to construct the recombinant vector pCA13-FasC. By lipofectamine (LF2000)-mediated transfection, pCA13-FasC was transfected into the 293 cells. RT-PCR and Western blot indicated that the murine soluble Fas C protein was expressed in the 293 cells. Apoptosis inducing test showed that the expression of this murine Fas C could block the Fas-induced apoptosis, which confirmed the biological activity of the recombinant Fas C.
Amino Acid Sequence ; Animals ; Animals, Newborn ; Base Sequence ; Cloning, Molecular ; DNA, Complementary ; genetics ; Fas Ligand Protein ; Gene Expression ; Membrane Glycoproteins ; biosynthesis ; genetics ; Mice ; Molecular Sequence Data ; Recombinant Proteins ; biosynthesis ; Sequence Analysis, DNA ; Sequence Homology, Amino Acid ; Transfection ; fas Receptor ; biosynthesis ; genetics

Objective To investigate the effect of ibuprofen on autophagy and astrocyte proliferation and their significances in rats with pentylenetetrazol (PTZ)-induced epilepsy. Methods Sixty male sprague-dawley rats were randomly divided into control group, PTZ group, 3-MA+PTZ group, ibuprofen+PTZ group and 3-MA+ibuprofen+PTZ group (n=12); activity of gamma-aminobutyric acid was blocked by PTZ to ignite epileptic rats in the latter 4 groups, and rats in the control group were intraperitoneally injected with normal saline once every one day; rats in the 3-MA+PTZ group and ibuprofen+PTZ group were given intraperitoneal injection of 3-MA (2.5 mg/kg) or ibuprofen (30 mg/kg) 30 min before PTZ injection;rats in the 3-MA+ibuprofen+PTZ group were given intraperitoneal injection of 3-MA (2.5 mg/kg)+ibuprofen (30 mg/kg) at the same time. The behavior and EEG features of rats were observed. Immunofluorescence staining and Western blotting were used to detect the expressions of microtubule-associated protein 1 light chain 3 (LC3) and glial fibrillary acidic protein (GFAP). Results (1) As compared with rats in the PTZ group and 3-MA+PTZ group, rats in the ibuprofen+PTZ group and 3-MA+ibuprofen+PTZ group had significantly decreased seizure grading and incidence of complete ignition, and significantly increased latency period (P<0.05). (2) The EEG waveform of the control group was normal; electroencephalogram of PTZ group and 3-MA+PTZ group showed sharp waves of high amplitude and spike waves; EEG wave peaks in the ibuprofen+PTZ group and 3-MA+ibuprofen+PTZ group decreased significantly, presenting frequent small spike waves and slow spike waves, among which ibuprofen+PTZ group showed more obvious changes. (3) The results of immunofluorescence staining and Western blotting showed that as compared with the PTZ group, 3-MA+PTZ group had statistically decreased LC3 expression and significantly increased GFAP expression (P<0.05); as compared with the PTZ group, ibuprofen+PTZ group had statistically increased LC3 expression and significantly decreased GFAP expression (P<0.05), however, 3-MA+ibuprofen+PTZ group had statistically decreased LC3 expression and significantly increased GFAP expression (P<0.05). Conclusion Ibuprofen can reduce astrocyte proliferation by promoting autophagy to affect seizures.

Objective:To evaluate the role of autophagy in morphine preconditioning-induced reduction of oxygen-glucose deprivation and restoration (OGD/R) injury in primary cortical neurons of mice and the relationship with c-Jun N-terminal kinase (JNK).Methods:Primary cortical neurons extracted from C57BL/6 neonatal mice within 24 h after birth were divided into 9 groups ( n=24 each) using a random number table method: control group (C group), OGD/R group, morphine preconditioning group (M group), autophagy inhibitor 3-methyladenine (3-MA) group (3-MA group), 3-MA+ morphine preconditioning group (3-MA+ M group), autophagy inhibitor chloroquine group (Ch group), chloroquine + morphine preconditioning group (Ch+ M group), JNK inhibitor SP600125 group (SP group) and SP600125 + morphine preconditioning group (SP+ M group). Morphine preconditioning: morphine was added at a final concentration of 3 μmol/L before OGD/R, and the cells were incubated for 2 h in OGD/R group. In 3-MA, Ch and SP groups, 3-MA 5 mmol/L, chloroquine 50 μmol/L and SP600125 25 μmol/L were added, respectively, and the cells were incubated for 150 min. In 3-MA+ M, Ch+ M and SP+ M groups, 3-MA 5 mmol/L, chloroquine 50 μmol/L and SP600125 25 μmol/L were added, respectively, at 30 min before morphine preconditioning. Then the cells were subjected to oxygen-glucose deprivation for 1 h followed by restoration of oxygen-glucose supply for 24 h. CCK-8 assay was used to detect the neuronal viability. The expression of JNK, phosphorylated JNK (p-JNK), microtubule-associated protein 1 light chain 3 (LC3), p62, Beclin1, caspase-3, and cleaved-caspase-3 was determined by Western blot. The autophagosomes and autolysosomes were counted using LC3-double fluorescent adenovirus transfection, and the neuronal apoptosis rate was determined by TUNEL staining. Results:Compared with C group, the neuronal viability was significantly decreased, the expression of Beclin1 was up-regulated, the expression of p62 was down-regulated, and the LC3Ⅱ/LC3Ⅰ ratio, p-JNK/JNK ratio, the number of autophagosomes and autolysosomes, cleaved-caspase-3/caspase-3 ratio and neuronal apoptosis rate were increased in OGD/R group ( P<0.001). Compared with OGD/R group, the neuronal viability, p-JNK/JNK ratio, LC3Ⅱ/LC3Ⅰ ratio and the number of autophagosomes and autolysosomes were significantly increased, the expression of Beclin1 was up-regulated, and the expression of p62 was down-regulated in M group, the LC3Ⅱ/LC3Ⅰ ratio was significantly decreased, and the expression of p62 was down-regulated in 3-MA group, the LC3Ⅱ/LC3Ⅰ ratio was significantly increased, and the expression of p62 was up-regulated in Ch group ( P<0.001), and no significant change was found in the parameters mentioned above in SP600125 group ( P>0.05). Compared with M group, the neuronal viability was significantly decreased, the LC3Ⅱ/LC3Ⅰ ratio was decreased, and the expression of p62 was up-regulated in M+ 3-MA group, the neuronal viability was significantly decreased, the LC3Ⅱ/LC3Ⅰ ratio was increased, and the expression of p62 was up-regulated in M+ Ch group, and the neuronal viability, LC3Ⅱ/LC3Ⅰ ratio and p-JNK/JNK ratio were significantly decreased, the expression of p62 was up-regulated, the number of autophagosomes and autolysosomes was decreased, the expression of Beclin1 was down-regulated, and the cleaved-caspase-3/caspase-3 ratio and neuronal apoptosis rate were increased in M+ SP group ( P<0.001). Conclusions:Morphine preconditioning can attenuate OGD/R injury by activating JNK, enhancing autophagy and inhibiting apoptosis in primary cortical neurons of mice.

Objective To observe the changes of iNOS, COX-2 and NF-κB in breast cancer MCF-7 cells treated with Yanghe decoction containing serum. Methods The MCF-7 human breast cancer cells were divided into blank group, low dose group, middle dose group and high dose group (4.5, 9, 18 g/kg, respectively). Real-time RT-PCR was used to detect the mRNA expression of iNOS and COX-2 at 24, 48, 72 and 96 hours, and Western-blot was used to detect the expression of NF-κB protein. Results Compared with the blank group, the expression of iNOS and COX-2 mRNA in the low dose group, middle dose group and high dose group decreased significantly at 24, 48, 72 and 96 hours(P<0.05), the the expression of NF-κB protein in the low dose group, middle dose group and high dose group decreased significantly at 24, 48, 72 and 96 hours (P<0.05). So it reduced with the increase of drug concentration and time. There was no significant difference in 72 hours after intervention. Conclusions The Yanghe decoction could reduce the expression of NF-κB, and then reducing the related inflammatory factors COX-2 and iNOS.