Objective To study the dynamic change of high-sensitivity C-reactive protein(hCRP),interleukin-1(IL-1?)interleukin-6(IL-6)in acute traumatic brain injury,and the correlation between the dynamic change and the scale of brain injury and the prognosis of the patients.Methods 192 cases patients were divided into different groups according to the Glasgow Coma Scale(GCS)and the Glasgow outcome Scale(GOS)three months later discharged from hospital respectively.The concentration of serum CRP,IL-1?,IL-6 were detected with immunoturbidimetry and ABC-ELISA 24 with in hours after traumatic brain injury and the concentration of serum after iniwn at ond day,three days and seven days was detected as well.Results Serum hCRP levels were remarkably elevated and reached peak value within 24 hours,and then gradually decreased.There were a negative correlation between hCRP,IL-1?,IL-6 and the patient's GCS groups(r=-0.162,P

Endoplasmic reticulum (ER) stress occurs in macrophage-rich areas of advanced atherosclerotic lesions and contributes to macrophage apoptosis and subsequent plaque necrosis. The purpose of the present study was to investigate the effects of caveolin-1 (Cav-1) on ER stress-induced apoptosis in cultured macrophages and the underlying mechanisms. RAW264.7 cells were incubated with thapsigargin (TG) to establish ER stress model. And Cav-1 expression was detected by Western blot. After being pretreated with filipin(III), a caveolae inhibitor, RAW264.7 cells were assayed with flow cytometry and confocal laser scanning microscopy to detect cell apoptosis. Moreover, p38 mitogen-activated protein kinase (MAPK) phosphorylation and C/EBP homologous protein (CHOP) expression were detected with Western blot. The results showed that Cav-1 expression was markedly increased at early stage of TG treatment (P < 0.05) and then decreased with prolonged or high dose TG treatments. The increasing of Cav-1 expression induced by TG in RAW264.7 cells was abolished under inhibition of caveolae by filipin(III) (P < 0.05). The effect of TG on apoptosis of RAW264.7 cells was further augmented after pretreatment with filipin(III) (P < 0.05). Western blotting showed that MAPK phosphorylation induced by TG was inhibited by filipin(III) in RAW264.7 cells (P < 0.05), whereas CHOP remained unchanged (P > 0.05). These results suggest that Cav-1 may play a critical role in suppressing ER stress-induced macrophages apoptosis in vitro, and one of the mechanisms may be correlated with the activation of p38 MAPK prosurvival pathway.
Animals ; Apoptosis ; drug effects ; Caveolin 1 ; genetics ; metabolism ; Cell Line ; Endoplasmic Reticulum Stress ; physiology ; Filipin ; pharmacology ; MAP Kinase Signaling System ; Macrophages ; cytology ; drug effects ; Mice ; Thapsigargin ; pharmacology ; Transcription Factor CHOP ; metabolism ; p38 Mitogen-Activated Protein Kinases ; metabolism

Objective To explore the value of Multi-detector CT in spinal cord angiography. Methods Ten patients with initial MR and clinical findings suggestive of spinal cord vessel disease were performed CT spinal cord angiography.Among these,7 patients were performed DSA later within 1 week, and 4 patients were therapy by operation.CT protocol:Toshiba Aquilion 64 slice CT scanner,0.5 mm thickness,0.5/r,120 kV,350 mA,choose aortic arch level as inspection position,and use"surestart" technique with CT threshold 180 HU.Contrast medium was Iohexol(370 mg I/ml),with injection velocity of 6 ml/s.The total volume was 80 ml.The CT spinal cord angiography images were analyzed according to disease model,disease range,feeding artery,fistula,draining veins,and were compared with DSA and operation results.Results All CT spinal cord angiography images displayed spinal vessel malformation. Among these,3 patients were inner-medullary arteriovenous malformation;2 patients were peri-medullary arteriovenous fistula;5 patients were spinal dural arteriovenous fistula.All cases showed disease range,and draining veins clearly,one patient had two vessels that were false positive,and all the other cases showed feeding arteries clearly,which were confirmed by DSA.Conclusion There are great values for CT spinal angiography in diagnosing spinal vessel disease,it can be a screening exam before DSA.

BACKGROUNDPolymorphisms of microRNA (miRNA), as a novel mechanism, are closely associated with disease states by interfering with miRNA function. Direct correlations have been identified between single-nucleotide polymorphisms (SNPs) in miRNA, but the effect on type 2 diabetes mellitus (T2DM) onset among Chinese population remains unclear. Therefore, the aim of this study was to identify correlations between common SNPs in miR-27a, miR-146a, and miR-124a with T2DM among a Chinese population, as well as to explore diabetic pathological mechanisms and the impact of environmental factors.

METHODSSNPscan technology was used to genotype 995 patients newly diagnosed with T2DM and 967 controls. Logistic regression analysis was performed to compare mutation frequencies between cases and controls.

RESULTSWe found no significant correlations between all genotypes of these miRNAs and T2DM in our research. However, stratification analysis identified a lower risk of T2DM associated with the rs531564GC genotype among younger subjects (age < 45 years) (adjusted P = 0.043; odds ratio [OR] = 0.73; 95% confidence interval [CI] = 0.54-0.99). Furthermore, the rs895819CC genotype in overweight people (24 ≤ body mass index [BMI] < 28) was significantly associated with an increased risk of T2DM (adjusted P = 0.042; OR = 1.73; 95% CI = 1.02-2.94), while the rs2910164 genotype in miR-146a was not significantly correlated with T2DM. The genetic risk score was calculated based on the number of risk alleles of the three SNPs and was found to be correlated to total cholesterol (adjusted P = 0.021).

CONCLUSIONSThe rs531564GC genotype acted as a protective factor to decrease the risk of T2DM in younger subjects (age < 45 years), while the presence of the rs895819CC genotype increased the risk of illness among overweight subjects (24 ≤ BMI < 28 kg/m 2 ). The presence of SNPs in miRNA might promote disease by affecting miRNA expression and gene function. Thus, miRNA mimics or inhibitors that directly regulate miRNA expression present novel and promising therapeutic targets.

Adult ; Aged ; Alleles ; Asian Continental Ancestry Group ; genetics ; Diabetes Mellitus, Type 2 ; genetics ; Female ; Genetic Predisposition to Disease ; genetics ; Genotype ; Humans ; Male ; MicroRNAs ; genetics ; Middle Aged ; Polymorphism, Single Nucleotide ; genetics ; Young Adult

Chemokine signal pathways are important for the regulation of tumour metastasis. Chemokine receptors CXCR4 (C-X-C chemokine receptor type 4) and XCR1 (chemokine XC receptor 1) are involved in the metastasis of breast cancer, while the interaction between them remains unclear. Here we first identified the interaction between CXCR4 and XCR1 based on membrane protein yeast two-hybrid assays. Bioluminescence resonance energy transfer (BRET) showed that XCR1 could competitively bind to CXCR4 to form a heterodimer (P < 0.01). Results of wound healing assays via transient transfection of XCR1 and CXCR4 into HEK293 cells showed that 41.55% of the migration area rate in the co-transformation group was lower than 58.75% in the CXCR4-alone group after adding 30 nmol/L S D F-β. The co-expression of XCR1 inhibited the cellular motility, possibly mediated by the SDF-1β (stromal cell-derived factor 1)/CXCR4 signal pathway (P < 0.05). Furthermore, CXCR4 on the cell surface after co-expression of XCR1 in CXCR4-EGFP transgenic HEK293 cells was detected by flow cytometry. And the result suggested that XCR1 could accelerate the internalization of CXCR4 into the heterodimer induced by 30 nmol/L SDF-1β (P<0.05), which increased the internalization rate from 14.38% to 64.10%. Finally, the phosphorylation of Akt and ERK, which were involved in cell proliferation and migration, respectively, were examined. After 10 minutes of SDF-1β stimulation, ERK phosphorylation in the CXCR4-alone group showed a 3.59-fold increase, whereas the increase of ERK phosphorylation in the co-transfected group was only 2.08-fold. Interestingly, heterodimer formation reduced the phosphorylation level of ERK and shortened the activation time, whereas the phosphorylation level of Akt remained unchanged. Collectively, our findings revealed the hetero-dimerization of CXCR4 and XCR1 and its effects on CXCR4-mediated cellular motility, receptor internalization, and ERK pathway phosphorylation. Therefore, XCR1-targeting drugs could be candidates for cross-desensitization of CXCR4 and might represent a possible option for inhibiting breast cancer metastasis.

OBJECTIVETo study the distribution of ScrF1 restriction polymorphism in intron 2 of the 3-hydroxy-3-methylglutaryl coenzyme A(HMG-CoA) reductase gene in Chinese Han population and the association of the polymorphism with coronary heart disease(CHD).

METHODSHMG-CoA reductase genotyping was performed using polymerase chain reaction-restriction fragment polymorphism.

RESULTSHMG-CoA reductase allelic frequencies of A, a were 0.519, 0.481; 0.440, 0.560 in CHD group and control group respectively. There was no significant difference in frequencies of allele and genotype in ScrF1 polymorphism between CHD group and control group(P>0.05). However, the levels of plasma very low density lipoprotein (VLDL) and TG in CHD patients with AA genotype were higher than those in CHD patients with other genotypes(P<0.05). The frequencies of A, a alleles at ScrF1 polymorphic site were significantly different from those reported in European Caucasians (0.44 vs 0.55, 0.56 vs 0.45, P<0.05).

CONCLUSIONNo direct association was found between the ScrF1 polymorphism and CHD, but there is a significant correlation between the AA genotype of the HMG-CoA reductase gene and the levels of plasma VLDL and TG in CHD group.

Adult ; Aged ; Aged, 80 and over ; Asian Continental Ancestry Group ; genetics ; Blood Chemical Analysis ; Cholesterol, VLDL ; blood ; Female ; Genetic Predisposition to Disease ; Humans ; Hydroxymethylglutaryl CoA Reductases ; genetics ; Hydroxymethylglutaryl-CoA-Reductases, NADP-dependent ; genetics ; Lipid Metabolism ; genetics ; Male ; Middle Aged ; Polymorphism, Genetic ; Triglycerides ; blood

OBJECTIVETo detect and clone mCD99L2 gene from mouse B lymphoma cell line A20 and construct its eukaryotic expression vector pcDNA3.1-mCD99L2.

METHODSThe expression of mCD99L2 mRNA in A20 cell line was detected by in situ hybridization. The total RNA of A20 cells was extracted to obtain the full-length cDNA of the coding region of mCD99L2 gene by RT-PCR, the product of which was ligated into pMD18-T vector and the DNA sequence of the insert was detected. The coding regions of mCD99L2 gene was amplified from pMD-mCD99L2 by PCR using primers containing EcoR I and Xho I sites and cloned into the eukaryotic expression vector pcDNA3.1/MycHis(+).

RESULTSIn situ hybridization identified positive expression of mCD99L2 gene in the A20 cell line. The full-length cDNA of mCD99L2 coding region of A20 cell line was obtained by RT-PCR, which yielded a product of 712 bp as expected, and the DNA sequence was completely homologus to the mCD99L2 cDNA reported in GenBank. Restriction endonuclease digestion and DNA sequencing indicated that the eukaryotic expression vector pcDNA3.1(+)- mCD99L2 had been constructed successfully.

CONCLUSIONmCD99L2 cDNA has been cloned from mouse B lymphoma cell line A20 and its eukaryotic expression vector pcDNA3.1(+)- mCD99L2 successfully constructed, which facilitates further functional study of mCD99L2 gene in mouse B lymphoma cell line A20.

12E7 Antigen ; Animals ; Antigens, CD ; biosynthesis ; genetics ; Base Sequence ; Cell Line, Tumor ; Cloning, Molecular ; DNA, Complementary ; chemistry ; genetics ; Eukaryotic Cells ; metabolism ; Gene Expression Regulation, Neoplastic ; Genetic Vectors ; genetics ; Lymphoma, B-Cell ; genetics ; pathology ; Mice ; Molecular Sequence Data ; RNA, Messenger ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, DNA

BACKGROUNDThe loss of cardiac myocytes is one of the mechanisms involved in acute myocardial infarction (AMI)-related heart failure. Autophagy is a common biological process in eukaryote cells. The relationship between cardiac myocyte loss and autophagy after AMI is still unclear. Carvedilol, a non-selective alpha1- and beta-receptor blocker, also suppresses cardiac myocyte necrosis and apoptosis induced by ischemia. However, the association between the therapeutic effects of carvedilol and autophagy is still not well understood. The aim of the present study was to establish a rat model of AMI and observe changes in autophagy in different zones of the myocardium and the effects of carvedilol on autophagy in AMI rats.

METHODSThe animals were randomly assigned to a sham group, an AMI group, a chloroquine intervention group and a carvedilol group. The AMI rat model was established by ligating the left anterior descending coronary artery. The hearts were harvested at 40 minutes, 2 hours, 24 hours and 2 weeks after ligation in the AMI group, at 40 minutes in the chloroquine intervention group and at 2 weeks in other groups. Presence of autophagic vacuoles (AV) in the myocytes was observed by electron microscopy. The expression of autophagy-, anti-apoptotic- and apoptotic-related proteins, MAPLC-3, Beclin-1, Bcl-xl and Bax, were detected by immunohistochemical staining and Western blotting.

RESULTSAVs were not observed in necrotic regions of the myocardium 40 minutes after ligation of the coronary artery. A large number of AVs were found in the region bordering the infarction. Compared with the infarction region and the normal region, the formation of AV was significantly increased in the region bordering the infarction (P < 0.05). The expression of autophagy- and anti-apoptotic-related proteins was significantly increased in the region bordering the infarction. Meanwhile, the expression of apoptotic-related proteins was significantly increased in the infarction region. In the chloroquine intervention group, a large number of initiated AVs (AVis) were found in the necrotic myocardial region. At 2 weeks after AMI, AVs were frequently observed in myocardial cells in the AMI group, the carvedilol group and the sham group, and the number of AVs was significantly increased in the carvedilol group compared with both the AMI group and the sham group (P < 0.05). The expression of autophagy- and anti-apoptotic-related proteins was significantly increased in the carvedilol group compared with that in the AMI group, and the positive expression located in the infarction region and the region bordering the infarction.

CONCLUSIONSAMI induces the formation of AV in the myocardium. The expression of anti-apoptosis-related proteins increases in response to upregulation of autophagy. Carvedilol increases the formation of AVs and upregulates autophagy and anti-apoptosis of the cardiac myocytes after AMI.

Adrenergic beta-Antagonists ; pharmacology ; Animals ; Apoptosis ; Apoptosis Regulatory Proteins ; analysis ; Autophagy ; drug effects ; Beclin-1 ; Carbazoles ; pharmacology ; therapeutic use ; Immunohistochemistry ; Male ; Microscopy, Electron, Transmission ; Myocardial Infarction ; drug therapy ; pathology ; Myocardium ; ultrastructure ; Propanolamines ; pharmacology ; therapeutic use ; Rats ; Rats, Wistar ; Vacuoles ; drug effects

OBJECTIVETo elucidate the roles of Toll-like receptor 3 (TLR3) on dendritic cells (DCs) in HBV infection.

METHODSPeripheral blood mononuclear cells (PBMCs) were isolated from 48 healthy volunteers (HV) and 50 chronically HBV-infected patients (CH). DCs were induced and proliferated in a culture medium with rhGM-CSF and rhIL-4. We stimulated DCs with poly I:C and then TLR3, HLA-DR, and CD86, and CD1a expressions were examined by flow cytometry at 0 h, 12 h, 24 h and 48 h. The mRNA expressions of TLR3 were quantified by real-time PCR.

RESULTSTLR3 expression on DCs before the poly I:C stimulation and afterwards on the 12 h, 24 h, and 48 h were 69.2%+/-20.4%, 76.0%+/-18.6%, 78.2%+/-19.5% and 85.5%+/-6.9% respectively in the CH group, and 70.8%+/-11.2%, 67.5%+/-20.9%, 86.3%+/-14.7%, 68.6%+/-16.9% in the HV group. The expressions of TLR3 were up-regulated significantly at 24 h and 48 h after stimulation with poly I:C in the HV group, and in the CH group they were not significantly increased at 24 h but obviously increased at 48 h. The mRNA expressions of TLR3 increased significantly at 12 h in the HV groups, and at 48 h in CH group. The rate of CD86 expressions increased after poly I:C stimulation, and the increased rates were 12.6%+/-9.8%, 23.8%+/-20.0%, 20.7%+/-14.3% in the CH group, and 31.0%+/-25.0%, 43.4%+/-24.7%, 44.6%+/-25.5% in the HV group at 12 h, 24 h and 48 h after poly I:C stimulation. There was a marked increase of the expression level of CD86 in the HV group. In contrast, the level was only slightly increased in the CH group (31.0% vs 12.6%). The differences between the two groups were significant at 24 h and 48 h. No significant differences were detected in HLA-DR and CD1a between the two groups.

CONCLUSIONSThe increase of expression level of TLR3 is slower in the CH group than that in the HV group. A marked increase of the expression level of CD86 is observed in the HV group. Our results suggest that abnormal expression of TLR3 and CD86 may relate to the persistence of HBV infection.

Adult ; B7-2 Antigen ; metabolism ; Dendritic Cells ; immunology ; metabolism ; Female ; Hepatitis B e Antigens ; blood ; Hepatitis B, Chronic ; blood ; Humans ; Male ; Middle Aged ; Toll-Like Receptor 3 ; metabolism ; Young Adult

Aim To observe the effects of microRNA-204/-211 deficiency on osteoarthritis(OA) induced by medial meniscus amputation (DMM) in mice. Methods 12 C57BL/6J wild-type (WT) mice were randomly divided into sham operation groups and DMM groups, namely WT-control group and WT + DMM group. And twelve microRNA-204/-211 gene knockout (miR-204/-211-dKO) mice were randomly divided into sham operation groups and DMM groups, namely dKO group, and dKO + DMM group. The pain sensitivity of mice was measured by the von Frey test before sacrificing. Three months after the operation, the mice were sacrificed. The knee joints and dorsal root ganglion (DRG) were taken for detection. The subchondral bone structure was detected by micro-CT. Sections of knee joint tissue were stained with toluidine blue, PCNA, type Ⅱ collagen and immunohistochemistry. DRG tissues were detected for related pain factors and inflammatory factors by RT-qPCR. Results Compared with the mice in the WT-Control group, mice in the WT + DMM group showed typical OA symptoms such as osteophyte formation, subchondral osteosclerosis, and decreased pain thresholds. The expression of collagen Ⅱ in cartilage significantly decreased, while the expression of MMP13 significantly increased. The expression of inflammatory and pain-related factors in DRG significantly increased. At the same time, the OA phenotypes of mice in dKO + DMM were more obvious than that of mice in the WT + DMM group, indicating that miR-204/-211 deficiency aggravated the OA induced by DMM in mice. In particular, DMM did not cause synovial hyperplasia and synovial inflammation in WT mice, which could not completely represent the pathological characteristics of OA patients in clinical practice. However, miR-204/-211 deficiency significantly promoted synovial hyperplasia and synovial inflammation of knee joints in DMM mice. Conclusions After DMM operation, miR-204/-211 deficient mice showed not only typical OA phenotypes such as osteophyte formation, subchondral osteosclerosis, cartilage destruction and lower pain threshold, but also synovial hyperplasia and synovitis, which could better represent the pathological characteristics of clinical OA patients. MiR-204/-211 deficient mice with DMM can be used as a new OA model and an ideal animal model for screening anti-OA drugs.