BACKGROUND: Arteriosclerosis obliterans (ASO) is a major cause of adult limb loss worldwide. Autophagy of vascular endothelial cell (VEC) contributes to the ASO progression. However, the molecular mechanism that controls VEC autophagy remains unclear. In this study, we aimed to explore the role of the GRB2 associated binding protein 1 (GAB1) in regulating VEC autophagy. METHODS: In vivo and in vitro studies were applied to determine the loss of adapt protein GAB1 in association with ASO progression. Histological GAB1 expression was measured in sclerotic vascular intima and normal vascular intima. Gain- and loss-of-function of GAB1 were applied in VEC to determine the effect and potential downstream signaling of GAB1. RESULTS: The autophagy repressor p62 was significantly downregulated in ASO intima as compared to that in healthy donor (0.80 vs. 0.20, t = 6.43, P < 0.05). The expression level of GAB1 mRNA (1.00 vs. 0.24, t = 7.41, P < 0.05) and protein (0.72 vs. 0.21, t = 5.97, P < 0.05) was significantly decreased in ASO group as compared with the control group. Loss of GAB1 led to a remarkable decrease in LC3II (1.19 vs. 0.68, t = 5.99, P < 0.05), whereas overexpression of GAB1 significantly led to a decrease in LC3II level (0.41 vs. 0.93, t = 7.12, P < 0.05). Phosphorylation levels of JNK and p38 were significantly associated with gain- and loss-of-function of GAB1 protein. CONCLUSION Loss of GAB1 promotes VEC autophagy which is associated with ASO. GAB1 and its downstream signaling might be potential therapeutic targets for ASO treatment.
Adaptor Proteins, Signal Transducing ; Adult ; Arteriosclerosis Obliterans/genetics* ; Autophagy ; GRB2 Adaptor Protein ; Humans ; Phosphoproteins/metabolism* ; Phosphorylation ; Protein Binding ; Signal Transduction

OBJECTIVESchimke immuno-osseous dysplasia (SIOD), is an autosomal recessive inherited disease caused by SMARCAL1 (MIM:20606622) mutations, while in about half of the patients no any mutation in SMARCAL1 could be found. This disease involves multiple systems and is characterized by short and dissymmetric stature with spondyloepiphyseal dysplasia, progressive renal failure, lymphopenia with recurrent infections, and hyperpigmented macules. This study aimed to analyze SMARCAL1 gene of 2 unrelated suspected SIOD children, to make definite diagnosis, and find more SMARCAL1 mutation types of Chinese SIOD.

METHODTwo suspected Chinese Han male SIOD children who visited our hospital from 2008 to 2014, aged 3 y 6 m and 7 y 8 m, both were short and had spondyloepiphyseal dysplasia, progressive renal failure, lymphopenia with recurrent infections. After informed consent, they and their parents's DNA were extracted from blood. PCRs for all 16 exons of SMARCAL1 were performed and PCR products were purified by 2% gel electrophoresis and sequenced directly. Pathogenicity of missense variations was confirmed by SIFT and sequencing SMARCAL1 of fifty normal controls.

RESULT(1) Four gene variations were found in the two children: Two reported missense mutations c.1129G>C, p.Glu377Gln and c.1933C>T, p. Arg645Cys. Two splicing mutations c.1334+1G>A and c.2142-1 G>A were detected. (2) c.1129G>C, p.Glu377Gln were reported as a disease-causing mutations before, but it was an single nucleotide polymorphism (SNP) which was found in 15 of 50 normal controls. (3) Two novel splicing mutations were found in this study: c.1334+1G>A and c.2142-1 G>A.

CONCLUSION(1) We detected 3 disease-causing mutations in 2 SIOD children by SMARCAL1 gene analysis, while 2 splicing mutations were novel mutations. (2) c.1129G>C, p.Glu377Gln was a SNP but not a disease-causing mutation at least in Chinese population.

Arteriosclerosis ; complications ; genetics ; Base Sequence ; Child ; Child, Preschool ; DNA Helicases ; genetics ; Exons ; Humans ; Immunologic Deficiency Syndromes ; complications ; genetics ; Lymphopenia ; Male ; Mutation ; Mutation, Missense ; Nephrotic Syndrome ; complications ; genetics ; Osteochondrodysplasias ; complications ; congenital ; genetics ; Polymorphism, Single Nucleotide ; Pulmonary Embolism ; complications ; genetics ; Renal Insufficiency

OBJECTIVETo investigate the relationship of the polymorphism of SG13S114A/T in ALOX5AP gene with atherosclerotic cerebral infarction (ACI).

METHODSBy polymerase chain reaction and restriction fragment length polymorphism, polymorphism of SG13S114A/T in ALOX5AP gene in 412 cases with ACI and 368 non-ACI controls were analyzed.

RESULTThere were no statistically significant differences in the ALOX5AP gene SG13S114 AA genotype and A allele frequencies between ACI group and control group (P>0.05).

CONCLUSIONThe results do not support genotype SG13S114 A allele as the risk gene for ACI.control group.

5-Lipoxygenase-Activating Proteins ; genetics ; Alleles ; Cerebral Infarction ; etiology ; genetics ; Female ; Genotype ; Humans ; Intracranial Arteriosclerosis ; complications ; genetics ; Male ; Polymorphism, Restriction Fragment Length

OBJECTIVETo study the protective effect of panax notoginseng saponins (PNS) on human umbilical vascular endothelial cells (HUVECs) injury induced by oxidized low-density lipoprotein (ox-LDL) for investigating the mechanism of PNS in treating arteriosclerosis obliterans (ASO).

METHODSTaking the cultured HUVECs as target cells, ox-LDL was used to establish a model of injured HUVEC and it was then intervened by PNS. The morphologic changes of HUVEC were observed under light microscope; activity of cells was determined by MTT method; the adhesive percentage between ox-LDL treated HUVEC and monocyte detennined by protein quantification and the protein expression of intercellular adhesion molecule-1 (ICAM-1) determined by flow cytometry.

RESULTSAt the time points of HUVEC being treated with ox-LDL (100 mg/L) for 12 h and 24 h, significant injury of HUVEC was shown, its activity reduced, the adhesion rate with monocytes elevated, and the protein expression of ICAM-l in HUVEC increased significantly (P < 0.05, P < 0.01). PNS showed significant effect in reversing all the above changes, as compared with the control group (without PNS treaded), respective significant difference was shown in all the four indexes (P < 0.05, P < 0.01).

CONCLUSIONPNS has a protective effect on endothelial cells injury induced by ox-LDL,which may be one of its mechanisms in treating ASO. The protective effect of PNS is probably by way of down-regulating the expression of ICAM-1 in endothelial cells and inhibiting the adherence of monocytes to endothelial cells.

Arteriosclerosis ; drug therapy ; metabolism ; physiopathology ; Cell Adhesion ; drug effects ; Cells, Cultured ; Drugs, Chinese Herbal ; pharmacology ; Endothelium, Vascular ; cytology ; drug effects ; metabolism ; Gene Expression ; drug effects ; Humans ; Intercellular Adhesion Molecule-1 ; genetics ; metabolism ; Lipoproteins, LDL ; metabolism ; Panax notoginseng ; chemistry ; Umbilical Veins ; cytology ; drug effects ; metabolism

OBJECTIVETo investigate the association of matrix metalloproteinase-3 (MMP-3) serum level and the promoter 5A/6A polymorphism of the MMP-3 gene with atherosclerotic cerebral infarction (ACI) in a Chinese Han population.

METHODSTwo hundred and fifteen patients with acute ACI from the Department of Neurology of Taizhou Hospital and 226 healthy controls were included in the study. Serum MMP-3 level was measured by enzyme-linked immunosorbent assay (ELISA). Genotype was determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) for the common 5A/6A functional promoter polymorphism of the MMP-3 gene.

RESULTSThe genotype distribution of the MMP-3 promoter 5A/6A polymorphism between the ACI patients group and the control group was significantly different (chi (2)= 9.389, P= 0.002). The frequencies of the 5A allele were 14.2% and 7.7% in the ACI patients group and the control group respectively (chi (2)= 9.430, P= 0.002). Serum level of MMP-3 in the ACI patients group was significantly higher than that in the control group (t= 24.867, P= 0.000). Among the ACI patients group, serum MMP-3 levels also had significant difference between the 5A/6A+ 5A/5A and the 6A/6A genotype (t= 2.057, P= 0.041).

CONCLUSIONThe present findings suggest that serum level of MMP-3 obviously increased within 48 hours of ischemic stroke and the genetic polymorphism of 5A/6A in the MMP-3 promoter is associated with ACI and MMP-3 expression in the Chinese Han population.

Adult ; Aged ; Aged, 80 and over ; Asian Continental Ancestry Group ; genetics ; Case-Control Studies ; Cerebral Infarction ; blood ; complications ; genetics ; Ethnic Groups ; genetics ; Female ; Gene Frequency ; Genotype ; Humans ; Intracranial Arteriosclerosis ; blood ; complications ; genetics ; Male ; Matrix Metalloproteinase 3 ; blood ; genetics ; Middle Aged ; Polymorphism, Genetic ; Promoter Regions, Genetic ; genetics

OBJECTIVETo investigate the effect of chronic enhanced external counterpulastion (EECP) on gene expression profiles of arterial endothelial cells (ECs) of pigs fed with high-cholesterol diet.

METHODSEight male pigs were fed with high-cholesterol diet for 12 weeks to induce arteriosclerosis and subjected to EECP for accumulative 36 h (2 h every other day for 18 sessions). Another 8 pigs on cholesterol-enriched diet and 6 normally fed pigs served as the arteriosclerosis model group and normal control group, respectively, and the high-cholesterol diet was maintained until the end of EECP treatment. The coronary artery was then isolated for transmission electro microscopy, and the abdominal aorta was observed using Sudan III staining. The gene expression profiles in ECs from the thoracic aorta using cDNA microarrays.

RESULTSMacrophages and foam cells were detected beneath the ECs in the coronary artery of pigs in the model group, but not in the other two groups. The ratios of Sudan III-positive area in the celiac aorta were significantly lower in normal control and EECP groups than in the model control group (P<0.05). Compared with the normal control group, the gene expressions of integrins-beta1 and CTGF were up-regulated in the model group. Compared with the model group, the expressions of integrins-beta1, CTGF and VCAM-1 were down-regulated and eNOS up-regulated in EECP group.

CONCLUSIONChronic EECP may reduce endothelial injury, down-regulate the gene expression level of integrin-beta1, CTGF and VCAM-1, lower cholesterol uptake and attenuate arterial endothelial inflammation to protect the pigs fed with high-cholesterol diet from arteriosclerosis.

Animals ; Aorta, Abdominal ; metabolism ; pathology ; Arteriosclerosis ; etiology ; genetics ; pathology ; Coronary Vessels ; metabolism ; pathology ; Counterpulsation ; methods ; Diet, Atherogenic ; Endothelial Cells ; metabolism ; Gene Expression Profiling ; Male ; Oligonucleotide Array Sequence Analysis ; methods ; Swine

OBJECTIVETo detect the correlation between the microsatellite DNA polymorphism of adrenomedullin(ADM) gene (repeated sequences of CA) and the atherosclerotic cerebral infarction (ACI).

METHODSWith PCR, ADM genotype was monitored from 189 normotensive subjects and 283 cerebral infarction patients. By using radioimmunoassay, their plasma ADM concentration was measured, so as the biochemical index.

RESULTSThe genotype distribution of ADM between the health control and ACI groups was significantly different, chi square was 28.732, P < 0.05. As one of the four alleles, including 11, 13, 14 and 19 alleles, the frequency of 19 allele in ACI groups was much higher than that in the health control group, chi square was 26.929, P < 0.05. However, there was no significant difference in plasma ADM concentration among the different genotypes of the ACI patients.

CONCLUSIONMicrosatellite DNA polymorphism of ADM gene may be associated with the genetic predisposition to ACI.

Adrenomedullin ; genetics ; Adult ; Aged ; Alleles ; Case-Control Studies ; Cerebral Infarction ; complications ; genetics ; Female ; Gene Frequency ; Genotype ; Humans ; Intracranial Arteriosclerosis ; complications ; genetics ; Male ; Microsatellite Repeats ; genetics ; Middle Aged ; Polymorphism, Genetic

OBJECTIVETo investigate the relationship between the polymorphism of apolipoprotein A5 gene (APOA5) -12238 T>C and atherosclerotic cerebral infarction (ACI).

METHODSThree hundred and forty-one subjects (170 ACI patients and 171 healthy controls) were collected to determine the genotypes by using polymerase chain reaction-restriction fragment length polymorphisms.

RESULTSAPOA5 allele frequencies of T/C were 0.588/0.412 and 0.424/0.576 in ACI group and control group respectively. There was significant difference in allele and genotype frequencies between ACI group and control group (P < 0.05). The levels of plasma triglyceride in ACI patients with TT genotype were higher than those in patients with CC genotypes (P < 0.05).

CONCLUSIONThe relationship is found between the site of APOA5 gene -12238 T>C and ACI. There is a significant correlation between TT genotype of APOA5 and the levels of plasma triglyceride in patients with ACI.

Apolipoprotein A-V ; Apolipoproteins A ; genetics ; Asian Continental Ancestry Group ; genetics ; Case-Control Studies ; Cerebral Infarction ; blood ; complications ; genetics ; Female ; Gene Frequency ; Genotype ; Humans ; Intracranial Arteriosclerosis ; blood ; complications ; genetics ; Lipids ; blood ; Male ; Middle Aged ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Polymorphism, Restriction Fragment Length

OBJECTIVETo analyze the role of resistin in insulin resistance (IR) through investigating the variation of plasma resistin levels and single-nucleotide polymorphisms (SNPs) in resistin gene 5' flanking region in stroke patients.

METHODSIn 103 atherothrombotic cerebral infarction (ACI) patients, 85 lacunar infarction (LI) patients, 70 intracerebral hemorrhage (ICH) patients, and 86 healthy controls, plasma resistin and insulin levels were measured by ELISA, SNPs in resistin gene 5' flanking region were detected by PCR and direct DNA sequencing. The subjects' body height and weight, the body mass index, quantitative insulin sensitivity check index (QUICKI), blood pressure, and the concentration of fasting plasma glucose, triglyceride, total cholesterol, creatinine, low-density lipoprotein, and high-density lipoprotein were also determined.

RESULTSQUICKI was significantly lower in the ACI and ICH patients (0.316 +/- 0.037 and 0.309 +/- 0.032, respectively) than that in the controls (0.342 +/- 0.043, P < 0.001), while plasma resistin level was significantly higher in the ACI and ICH patients (6.36 +/- 3.79 and 7.15 +/- 4.27 ng/mL, respectively) than that in the controls (5.28 +/- 2.56 ng/mL, P < 0.05), but such difference was not observed in the LI patients compared with controls. There was a statistically negative correlation between plasma resistin level with QUICKI (r = -0.228, P < 0.001). The distributions of allele and genotype frequencies of resistin gene - 420C > G and - 537A > C SNPs were not significantly different among the different groups, and those SNPs were not correlated with other clinical and biochemical parameters.

CONCLUSIONSPlasma resistin is associated with stroke by participating in the development of IR. The SNPs in resistin gene 5' flanking region has no impact on the plasma resistin level.

Adult ; Aged ; Aged, 80 and over ; Blood Glucose ; metabolism ; Cerebral Infarction ; blood ; genetics ; Creatinine ; blood ; Female ; Humans ; Insulin ; blood ; Insulin Resistance ; physiology ; Intracranial Arteriosclerosis ; blood ; genetics ; Lipoproteins ; blood ; Male ; Middle Aged ; Polymorphism, Single Nucleotide ; Resistin ; blood ; genetics ; Stroke ; blood ; genetics ; Triglycerides ; blood

Taxol is one of the most potent chemotherapeutic agents known, showing excellent activity against a range of cancers. In addition to anticancer, taxol has the effect of preventing graft arteriosclerosis, antiscaring formation and inhibiting angiogenesis. There are five possible routes to industrialize taxol production: isolation from the bark of the yew species, total synthesis, semisynthesis, tissue or cell culture, endophytic fungal fermentation and metabolism engineering. There are at least 14 genes related to the taxol biosynthesis had been cloned from yews and functionally expressed in different hosts. The combinational expression system of taxol makes progress as the clarification of biosynthetic pathway of taxol.
Antineoplastic Agents, Phytogenic ; biosynthesis ; chemical synthesis ; chemistry ; isolation & purification ; Arteriosclerosis ; prevention & control ; Genes, Plant ; Genetic Engineering ; Neoplasms ; drug therapy ; Paclitaxel ; biosynthesis ; chemical synthesis ; chemistry ; isolation & purification ; Taxoids ; chemical synthesis ; chemistry ; Taxus ; chemistry ; genetics