OBJECTIVETo explore the association of 8-hydroxyguanine glycosidase OGG1 Ser326Cys polymorphism with semen quality and the risk of male infertility.

METHODSThis case-control study included 620 idiopathic infertile patients and 385 normal fertile controls. We determined their genotypes by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and analyzed their semen quality by computer-aided semen analysis (CASA).

RESULTSThe individuals with OGG1 326 Cys/Cys showed significantly lower sperm motility and concentration ([52.1 +/- 26.7]% and (3.75 +/- 0.91) x 10(6)/ml, ln transformed value) than the Ser/Ser carriers ([59.0 +/- 21.8] % and (4.12 +/- 0.88) x 10(6)/ml, ln transformed value) (P < 0.05). The risk of male infertility increased 69% in the OGG1 326Cys allele carriers as compared with the Ser carriers (OR = 1.69, 95% CI: 1.24 -2.31).

CONCLUSIONOGG1 326 Ser/Cys polymorphism might contribute to the risk of male infertility in the southern Chinese population.

Adult ; Case-Control Studies ; DNA Glycosylases ; genetics ; Genotype ; Humans ; Infertility, Male ; genetics ; Male ; Middle Aged ; Polymorphism, Single Nucleotide ; Semen Analysis ; Young Adult

AlM: To investigate the relationship between the anterior ischemic optic neuropathy ( AlON ) and the carotid artery change using doppler ultrasound.METHODS:Fifty-four cases of AlON patients and 54 cases of healthy control were observed, atherosclerotic spots were detected by the application of color ultrasound.RESULTS:ln AlON group of 54 patients, 38 cases appeared carotid atherosclerosis, accounting for 70%. The number of cases with hard plaque, soft plaque and mixed plaques were 18, 13, and 7 respectively, accounting for 33%, 24% and 13%. ln the control group, 20 cases were detected atherosclerotic change, accounting for 37%. And the number of cases with hard plaque, soft plaque and mixed plaques were 12, 5 and 3 respectively, accounting for 22%, 9%, 6%. Significant stenosis and velocity change were showed in neither AlON group nor control group. Compared with the control group, AlON group had more cases of atherosclerotic plaque, the difference was statistically significant (χ2=12. 836, P=0. 005)CONCLUSlON: The incidence of AlON is correlated with carotid atherosclerosis, and carotid ultrasonography is significantly valuable for AlON etiology and diagnosis.

The present study aimed to investigate the effects of interleukin-1β (IL-1β) at different doses on collagen synthesis and decomposition in cultured cardiac fibroblasts from neonatal Sprague-Dawley rat. Cardiac fibroblasts were treated with IL-1β (0.01, 0.1, 1, 10, 100 ng/mL) for 24 h. Cell DNA synthesis was measured by (3)H-thymidine ((3)H-TdR) incorporation and collagen synthesis was measured by (3)H-proline ((3)H-Pro) incorporation. Matrix metalloproteinase (MMP) activity was measured by gelatinase zymography. MMP-2, MMP-9 protein expressions were measured by Western blot. mRNA expressions of MMP-2 and MMP-9 were detected by reverse transcription-polymerase chain reaction (RT-PCR). Compared with that in the control group, the incorporation of (3)H-TdR and (3)H-Pro decreased in high-dose IL-1β groups (≥0.1 ng/mL) but not in low-dose IL-1β group (0.01 ng/mL). IL-1β significantly increased MMP-2 and MMP-9 activities. IL-1β (0.01-100 ng/mL) also dose-dependently increased the protein and mRNA expressions of MMP-2 and MMP-9 (P<0.05, P<0.01), respectively. These results suggest that IL-1β decreases collagen synthesis and MMP activities through transcriptional and posttranslational processes via degrading collagen in a dose-dependent way. Elevation of IL-1β is possibly involved in the process of ventricular remodeling after myocardial infarction, and the concentration of IL-1β is possibly a major factor which affects the extent of ventricular remodeling.
Animals ; Cells, Cultured ; Collagen ; biosynthesis ; Fibroblasts ; cytology ; metabolism ; Interleukin-1beta ; pharmacology ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Myocardial Infarction ; Myocardium ; cytology ; Rats ; Rats, Sprague-Dawley ; Ventricular Remodeling

Adjuvants, Immunologic ; pharmacology ; Animals ; Anti-Inflammatory Agents, Non-Steroidal ; pharmacology ; Antiviral Agents ; pharmacology ; Caffeic Acids ; chemistry ; isolation & purification ; pharmacology ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Echinacea ; chemistry ; classification ; Fatty Acids, Unsaturated ; isolation & purification ; pharmacology ; Molecular Structure ; Plants, Medicinal ; chemistry ; Polyunsaturated Alkamides

OBJECTIVETo study the lipid constituents from Echinacea purpurea.

METHODThe compounds were isolated by chromatography method and the structures were identified on the basis of spectral analyses.

RESULTFive compounds were isolated and identified as, 1 beta, 6 alpha-dihydroxy-4(14)-eudesmene(1), (2E, 4E, 8Z, 10E)-N-isobutyl-2,4,8,10-dodecatetraenamide(2), (2E, 4E, 8Z, 10Z)-N-isobutyl-2,4,8,10-dodecatetraenamide(3), cerotic acid(4), hyxacosyl alcohol(5).

CONCLUSIONCompounds 1,4 and 5 were obtained from the plant for the first time.

Echinacea ; chemistry ; Fatty Acids ; chemistry ; isolation & purification ; Fatty Alcohols ; chemistry ; isolation & purification ; Plants, Medicinal ; chemistry ; Sesquiterpenes ; chemistry ; isolation & purification

BACKGROUND: Recent clinical observation reported that there was a significant correlation between change in circulating vascular endothelial growth factor (VEGF) levels and the occurrence of severe acute graft-versus-host disease (GVHD) following allogeneic hematopoietic stem cell transplantation (allo-HSCT), but the action mechanisms of VEGF in GVHD have not been demonstrated. METHODS: This study investigated whether or not blockade of VEGF has an effect on acute GVHD in a lethally irradiated murine allo-HSCT model of B6 (H-2b)-->B6D2F1 (H-2b/d). Syngeneic or allogeneic recipient mice were injected subcutaneously with anti-VEGF peptides, dRK6 (50 microg/dose) or control diluent every other day for 2 weeks (total 7 doses). RESULTS: Administration of the dRK6 peptide after allo-HSCT significantly reduced survival with greaterclinical GVHD scores and body weight loss. Allogeneic recipients injected with the dRK6 peptide exhibited significantly increased circulating levels of VEGF and expansion of donor CD3+ T cells on day +7 compared to control treated animals. The donor CD4+ and CD8+ T-cell subsets have differential expansion caused by the dRK6 injection. The circulating VEGF levels were reduced on day +14 regardless of blockade of VEGF. CONCLUSION: Together these findings demonstrate that the allo-reactive responses after allo-HSCT are exaggerated by the blockade of VEGF. VEGF seems to be consumed during the progression of acute GVHD in this murine allo-HSCT model.
Animals ; Body Weight ; Graft vs Host Disease ; Hematopoietic Stem Cell Transplantation ; Hematopoietic Stem Cells ; Humans ; Mice ; Oligopeptides ; Peptides ; T-Lymphocyte Subsets ; T-Lymphocytes ; Tissue Donors ; Vascular Endothelial Growth Factor A

OBJECTIVETo construct a small interfering RNA (siRNA) targeting mouse vascular endothelial growth factor receptor-2 (VEGFR2) and study its serum stability and gene silencing efficiency in vitro.

METHODSThe synthesized siRNA targeting VEGFR2 (siVEGFR2) diluted in RNase-free water was mixed at a 1:1 ratio with fresh serum and incubated at 37 degrees celsius;. Gel electrophoresis was performed to determine the integrity of siVEGFR2 incubated for different time lengths. Oligofectamine 2000 was used to mediate siVEGFR2 transfection of MS1 cells, and semi-quantitative RT-PCR was used to evaluate VEGFR2 gene silencing effect induced by the siRNA.

RESULTSThe naked siRNA incubated in serum underwent gradual degradation with prolonged incubation time and became virtually undetectable after 24 h. Transfection of MS1 cells with siVEGFR2 significantly down-regulated the expression of VEGFR2 mRNA in comparison with the blank group and control siRNA transfection group (P<0.001), while no significant difference was found in VEGFR2 mRNA levels between the latter two groups (P=0.157).

CONCLUSIONThe naked siRNA is unstable in serum and not suitable for direct administration in vivo. The designed siRNA can effectively silence the VEGFR2 gene expression in MSI cells in vitro.

Animals ; Cell Line, Tumor ; Mice ; Neovascularization, Pathologic ; genetics ; RNA Interference ; RNA, Small Interfering ; genetics ; Transfection ; Vascular Endothelial Growth Factor Receptor-2 ; genetics

OBJECTIVETo evaluate the effect of sustained-release alpha-lipoic acid tablets (SRLA) on blood lipid, glucose and insulin levels in hyperlipidemic New Zealand rabbits.

METHODSTwenty-four New Zealand rabbits were randomized into normal diet group, high-fat diet group, and high-fat diet + SRLA (300 mg/tablet) group with corresponding feed. At the beginning and 4 weeks after the feeding, the serum levels of total cholesterol (TC), triglycerides (TG), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), blood glucose, and serum insulin were measured, and insulin sensitivity index (ISI) was calculated.

RESULTSFour weeks after feeding with high-fat diet, the insulin levels was elevated and the ISI lowered in the New Zealand rabbits, indicating successful establishment of the animal model of hyperlipidemia. Compared with the high-fat diet group, the serum levels of TG, TC, LDL-C and insulin were significantly reduced (P<0.05), and the ISI was significantly increased (P<0.05) in high fat diet + SRLA group. But no statistically significant difference was found in the blood glucose among the 3 groups.

CONCLUSIONSRLA can significantly correct blood lipid and insulin disorders in hyperlipidemic New Zealand rabbits and prevent the occurrence of insulin resistance and hyperlipidemia.

Animals ; Blood Glucose ; metabolism ; Delayed-Action Preparations ; Hyperlipidemias ; blood ; drug therapy ; metabolism ; Insulin ; metabolism ; Lipids ; blood ; Male ; Rabbits ; Tablets ; Thioctic Acid ; administration & dosage ; pharmacology ; therapeutic use

OBJECTIVEThe loss of caspase-8 expression correlates with unfavorable survival outcomes in neuroblastoma (NB). Caspase-8 gene inactivation is caused by methylation. This study aimed to explore the effect of the demethylation agent 5-azacytidine on caspase-8 expression and whether 5-azacytidine can increase the sensitivity of chemotherapy drug doxorubicin to NB cells.

METHODSCaspase-8 mRNA expression in NB cell lines (SH-SY5Y cells) was examined by RT-PCR before and after 5-azacytidine treatment. Survival rates of SH-SY5Y cells were detected using MTT analysis and compared among the doxorubicin alone treatment, 5-azacytidine along with doxorubicin treatment, and caspase-8 inhibitor+5-azacytidine+doxorubicin treatment groups.

RESULTSCaspase-8 mRNA was not expressed in untreated SH-SY5Y cell lines. Caspase-8 mRNA expression in SH-SY5Y cells was detectable 3 days after 5-azacytidine treatment, and increased significantly 5 days after 5-azacytidine treatment (P < 0.05). Survival rates of SH-SY5Y cells treated with 5-azacytidine along with different concentrations of doxorubicin (0.05, 0.1,0.25, 0.5 microg/mL) were (77.61 +/- 7.30)%, (57.35 +/- 6.64)%, (46.25 +/- 4.46)% and (35.59 +/- 5.12)%, respectively, which were significantly lower than those treated with doxorubicin alone (94.89 +/- 4.15%, 80.60 +/- 8.50%, 64.48 +/- 4.92% and 52.32 +/- 6.71%) (P < 0.01). Caspase-8 inhibitor pretreatment resulted in an increased survival rate of SH-SY5Y cells (92.95 +/- 3.48%, 78.39 +/- 4.28 %, 62.31 +/- 6.50% and 49.92 +/- 5.77%) compared with the 5-azacytidine+doxorubicin treatment group.

CONCLUSIONS5-azacytidine may enhance anti-tumor efficacy of doxorubicin to NB cell lines, possibly through an up-regulation of caspase-8 mRNA expression.

Antineoplastic Agents ; pharmacology ; Azacitidine ; pharmacology ; Caspase 8 ; genetics ; Caspase Inhibitors ; Cell Line, Tumor ; Doxorubicin ; pharmacology ; Drug Synergism ; Humans ; RNA, Messenger ; analysis

The aim of this paper is to report the study on gene silencing efficiency of siRNA targeted against mouse VEGFR2 (siVEGFR2) in vitro mediated by polyethyleneimine (PEI) and its anti-tumor effect in vivo. CY3-labeled siRNA was compounded into PEI and transfected into MS1 cells. Confocal microscopy was used to image the subcellular distribution of siRNA in MS1 cells. Semi-quantitative RT-PCR and Western blotting were used to evaluate VEGFR2 gene silencing induced by siVEGFR2/PEI complexes. A tumor-bearing nude mice model was established to compare the anti-tumor effect after delivered by local and systemic routes. siVEGFR2/PEI complex-transfected cells exhibited much fluorescence in cytoplasm with no evidence of nuclear accumulation. The expression levels of VEGFR2 mRNA and protein in PEI-transfected cells were significantly down-regulated compared with that in blank group, the silencing efficiency were 28.2% and 23.6% respectively. The tumor sizes in mice intratumorally injected with siVEGFR2/PEI complexes (189.429 +/- 17.562 mm3) were reduced definitely compared to that in mice injected with siVEGFR2/PEI complexes via vein route (315.507 +/- 20.491 mm3), or to saline groups (365.844 +/- 20.713 mm3). The study demonstrated that PEI could effectively transfect siRNA into cells and silence the VEGFR2 gene expression. Intratumoral delivery is more suitable for non-targeted modified PEI/siRNA complexes to inhibit the tumor growth in vivo. The present data lay a solid foundation to further study on the gene silencing mechanism for PEI-medicated RNAi and its anti-tumor efficiency in vivo.
Animals ; Cell Line, Tumor ; Cells, Cultured ; Endothelial Cells ; cytology ; metabolism ; Female ; Gene Silencing ; Humans ; Lung Neoplasms ; metabolism ; pathology ; Mice ; Mice, Nude ; Neoplasm Transplantation ; Polyethyleneimine ; chemistry ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; genetics ; metabolism ; Spleen ; cytology ; Transfection ; Tumor Burden ; Vascular Endothelial Growth Factor Receptor-2 ; genetics ; metabolism