Detecting polymorphisms in MMP genes.
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Autoradiography↗
Biomedical subjects
S Ye
Publications and source records attributed to S Ye.
[Study of the cytotoxity against human hepatocellular carcinoma cells induced by the MAGE-1 gene modified dendritic cells].
OBJECTIVE: To genetically modify dendritic cells (DC) with a tumor associated antigen gene MAGE-1 and to observe in vitro the cytotoxic effect induced by this genetically modified DC against human hepatocellular carcinoma (HCC) cell line SMMC7721, thus giving a primary verification of the possibility of this genetically modified DC to induce specific antitumor immunity against HCC and serving as a new type of vaccine. METHODS: The MAGE-1 gene was inserted into the retrovirus vector LXSN to construct the recombinant retrovirus LMSN. The monocyte-derived DCs were transfected by LMSN and control virus LXSN respectively as well as a third group was set up as non-transfected control. The MAGE-1 gene expression in LMSN transfected DC was identified by Western blot and the in vitro cytotoxities against SMMC7721 induced by three groups of DC were tested by MTT assay. RESULTS: A recombinant retrovirus LMSN containing the tumor rejection antigen gene MAGE-1 was successfully constructed and the MAGE-1 gene expression in LMSN transfected human monocyte-derived DC was induced. The LMSN transfected DC could induce very high lytic activity against SMMC7721 in vitro with the lytic activity of 78.9%+/-3.6%. Nevertheless, the LXSN transfected DC and non-transfected DC only induced relatively lower lytic activity as 34.7%+/-4.3% and 3.9%+/-2.0%, respectively. The difference of the lytic activities between those three groups as statistically significant (P<0.01). CONCLUSIONS: The MAGE-1 gene transfected DC can induce higher in vitro cytotoxicity against SMMC7721, suggesting the possibility of this genetically modified DC to induce specific antitumor activity and to serve as a new type of vaccine for HCC.
Animals↗
[Effects of interferon-alpha on recurrence and metastasis of hepatocellular carcinoma after curative resection in nude mice].
OBJECTIVE: To study the effect of interferon-alpha (IFN-alpha) on the recurrence and metastasis of hepatocellular carcinoma (HCC) in nude mice, and to clarify if there is synergistic effect treated by combination of IFN-alpha and all-trans retinoic acid (ATRA). METHODS: The effect of IFN-alpha and/or ATRA on the proliferation of HCC cell lines was measured in vitro. The metastatic model of human HCC in nude mice LCI-D20 was used in present study. Curative resection was performed at 10th day after implantation in 44 nude mice. Drugs were given at the next day after resection. IFN-alpha was administered subcutaneously at doses of 3+/-10(5) U/day, 6+/-10(5) U/day, respectively. ATRA was administered p.o. at a dose of 20 mg/kg/day, and IFN-alpha 6+/-10(5)U/day combined with ATRA 20 mg/kg/day. The mice were sacrificed 35 days after treatment. The recurrent tumor size was measured and the presence of intrahepatic dissemination and lung metastases were recorded. RESULTS: The effect of IFN-alpha and/or ATRA on the proliferation of HCC cells SMMC7721, BEL-7402, BEL-7405, and MHCC97 was not obvious. The combination of IFN-alpha and ATRA had no synergistic effect in vitro. The lung metastatic rate, the liver recurrent rate, the size of main recurrent lesions, the number of intrahepatic disseminating nodules and the largest disseminating nodule of the controlled group was 100%(12/12), 100%(12/12), (1346.3+/-4.2 ) mm(3), 8.2+/-4.4, 864 mm(3), respectively; whereas it was 0, 87.5%(7/8), (8.7+/-2.9) mm(3), 2.3+/-0.6, and 7.8mm(3), respectively in the IFN-alpha 3+/-10(5) U/day treated group(P<0.05); 0, 12.5%(1/8), 0.5mm(3), 2, 0.5 mm(3) in the IFN-alpha 6+/-10(5)U/day treated group(P<0.05); 0, 12.5%(1/8), 1 mm(3), 2.5+/-0.7, 8 mm(3) in the IFN-alpha 3+/-10(5) U/day and ATRA treated group(P<0.05); 87.5%, 100%, (1472.6+/-5.6) mm(3), 7.3+/-3.8, 768.5 mm(3) in the ATRA treated group (P>0.05). CONCLUSIONS: IFN-alpha has an inhibitory effect on intrahepatic recurrence and lung metastasis of human HCC after curative resection in nude mice, and the effect is enhanced with increasing dose. IFN-alpha and ATRA have no synergistic effect according to in vivo and in vitro test. ATRA has no effect on recurrence and metastasis of HCC.
Animals↗
[Microsurgical treatment of foramen magnum tumors].
OBJECTIVES: To review the radiological features, and results of microsurgical treatment of foramen magnum tumors. METHODS: We retrospectively analyzed the results of microsurgical treatment of 31 patients with foramen magnum tumor. Fifteen patients had pathological schwannomas (neurinomas), 13 patients meningiomas, and 3 patients chordomas. According to tumor extension, MRI findings and microsurgical approaches, the tumors were classified in two types. Type I tumors located at the ventral foramen magnum, including those originated from the posterior fossa with minimal enlargement of the ventral or anterior-lateral (15 patients). Type II tumors located at the posterior or lateral foramen magnum, including those arising from the spinal canal with minimal enlargement of the foramen magnum (16 patients). Surgical approaches to tumors in this series included suboccipital posterior approach (18 patients), extreme lateral or transcondylar approach (10), and suboccipital retrosigmoid approach (3). RESULTS: Total removal was achieved in 25 patients, subtotal removal in 5, and partial removal in 1. No operative death occurred. Type I foramen magnum tumors were totally removed in 9 patients, subtotally removed in 5 and partially removed in 1. All of type II foramen magnum tumors were totally removed. CONCLUSIONS: To totally resect type I foramen magnum tumors, extreme lateral transcondylar approach is an optimal choice. Type II foramen magnum tumors can be totally removed with good prognosis.
Adolescent↗
[p53 and vascular endothelial growth factor expression in astrocytoma and their relation to angiogenesis].
OBJECTIVE: This investigation was done to show the pathological importance of p53 protein detection, vascular endothelial growth factor expression (VEGF) and intratumoral microvessel density (IMVD) in astrocytoma. METHODS: The surgical specimens from 60 brain astrocytoma patients were stained immunohistochemically for p53 and IMVD expressions. IMVD was calculated by labeling the endothelial cells of the blood vessels within the tumor. RESULTS: p53, VEGF expression were closely correlated with histopathological grade of astrocytoma. Positive p53 protein accumulation and VEGF expression were found in 45.0% (27/60) and 65.0% (39/60) of tumors. The conformation rate of p53 and VEGF was 70.0% (42/60). IMVD was significantly higher in the p53-positive or VEGF-positive tumors than in the negative ones (P < 0.05). Comparing the IMVD in p53+/VEGF+ group, p53-/VEGF+ group and p53+/VEGF- group, p53-/VEGF- group, its difference was also highly significant (P < 0.05). CONCLUSION: (1) p53 protein detection, VEGF expression and IMVD can be considered as a biological indicator of malignant potential in brain astrocytoma patients and (2) p53 and VEGF expressions, both contributing to the tumor neovascularization, may be helpful in the understanding of intra-tumoral angiogenetic mechanism in the future.
Adolescent↗
[Transcatheter arterial chemoembolization for postoperative residual tumor of hepatocellular carcinoma].
OBJECTIVE: To understand and analyze the survival and prognostic factors of postoperative residual tumor of hepatocellular carcinoma treated by transcatheter arterial chemoembolization. METHODS: Transcatheter arterial chemoembolization was performed in 74 patients who were identified as having residual lesions by ultrasonography, hepatic arterial angiography or enhanced computed tomography about two months after resection of hepatocellular carcinoma. Kaplan-Meier method was used for survival and Cox regression model for prognostic factors. RESULTS: The 1-, 2- and 3-year survival rates were 78.0%, 57.6%, 37.0%, with a median survival of 33 months. Univariate analysis indicated that a primary tumor over > 5 cm in diameter, vascular involvement by the primary tumor and TNM extent of the residual lesion were important factors indicating a bad prognosis, where as the combination of other treatment methods such as percutaneous ethanol injection (PEI), and/or radiotherapy indicated a better prognosis. However, multivariate analysis showed that vascular involvement by the primary tumor and the other combined local treatments were independent factors of prognosis. CONCLUSION: Transcatheter arterial chemoembolization is effective in treating the postoperative residual tumor of hepatocellular carcinoma. Further improvement is observed if combined with other local therapies such as percutaneous ethanol injection or radiotherapy.
Adolescent↗
[Serum apolipoprotein A I, B100 and E levels and apolipoprotein E polymorphism in patients with Alzheimer's disease and multiple infarction dementia in Chinese population].
OBJECTIVE: To investigate the relationship of serum apolipoprotein (apo) A I, B100 and E levels and apo E polymorphism to Alzheimer's disease (AD) and multiple infarction dementia (MID) in Chinese population. METHODS: The apoE phenotypes were assayed by isoelectric focusing and immunoblotting, and apolipoproteins were determined by radial immunodiffusion assay in 75 patients with AD, 36 patients with MID and 60 control subjects. RESULTS: The frequency of apo E4 allele (epsilon 4) was significantly higher and the frequency of epsilon 2 allele was lower in AD group, compared with those in the control group (0.2333 vs 0.0666, 0.0467 vs 0.0833, P < 0.05). The frequency of epsilon 4 allele in MID group was also higher than that in the control group (0.2083 vs 0.0666, P < 0.05). The fasting serum apoA 1 and E levels in AD and MID groups were remarkably lower than those in the control group (P < 0.001 and P < 0.05). CONCLUSION: Apo E4 allele epsilon 4 was associated with AD and MID. ApoE4 might be a risk factor for AD and MID, and apo E2 might be a protective factor for AD. The serum apo A I and apoE levels were significantly decreased in patients with Alzheimer's disease and multiple infarction dementia.
Aged↗
[Determination of salicin in extract of willow bark by high performance liquid chromatography].
An HPLC method for the determination of salicin in extract of willow bark is described. Chromatographic analysis was carried out on a Kromasil C18, 5 microns column(4.6 mm i.d. x 250 mm) with methanol-0.01 mol/L KH2PO4 buffer (pH 4.01) (15:85, volume ratio) as mobile phase. The detection wavelength was 265 nm. Salicin was extracted from samples with methanol-water(50:50, volume ratio), and centrifuged. Ten microL of supernatant were injected. The average recoveries were from 96.1% to 101.2% (n = 5), and the relative standard deviation (RSD) was 1.43%. The method is simple, rapid and accurate.
Benzyl Alcohols↗
[Effects of low frequency pulsed electric field on insulin studied by fluorescent spectrum].
Insulin, an intercellular signal molecular, plays a critical role in transferring life information. The studies on effects of pulsed electric fields on insulin molecular are meaningful for explaining the mechanism of biological effects of electromagnetic fields. The experiment results demonstrate that the conformation of insulin molecular has been altered by electric fields exposure for 30 minutes or by heat exposure for 50 hours. The results suggest that pulsed electric field exposure as heat exposure will after conformation of insulin molecular by breaking some hydrogen bonds and recombining another hydrogen bonds. The intercellular signal molecular may be an important target of electromagnetic fields.
Electricity↗
Switching of stereochemistry. Dramatic effect of HMPA on the stereoselectivity of the cyclopropanation reaction of telluronium allylides with alpha,beta-unsaturated esters and amides.
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Allyl Compounds↗
Allele-specific regulation of matrix metalloproteinase-12 gene activity is associated with coronary artery luminal dimensions in diabetic patients with manifest coronary artery disease.
Both the processes of atherosclerosis and plaque rupture are indicated to be influenced by matrix metalloproteinase (MMP) activity. We therefore searched for common functional variation in the matrix metalloelastase (MMP-12) gene locus that may be implicated in coronary artery disease. Single-strand conformation polymorphism analysis of DNA from healthy individuals detected a common polymorphism within the MMP-12 gene promoter (an A-to-G substitution at position -82). The frequency of the G allele was 0. 19. The polymorphism influences the binding of the transcription factor activator protein-1 (AP-1) in electromobility shift assay. A higher binding affinity of AP-1 to the A allele was associated with higher MMP-12 promoter activity in vitro in transient transfection studies in U937 and murine lung macrophage (MALU) cells. Phorbol 12-myristate 13-acetate (PMA) and insulin, 2 known activators of AP-1, increased the binding of AP-1 to the MMP-12 promoter, with higher affinity for the A allele. In transfection experiments, both the A and the G alleles responded to insulin and PMA, the A allele showing higher promoter activity than the G allele. Furthermore, Western blot analysis demonstrated that insulin increased MMP-12 protein production. To analyze whether the -82 A/G polymorphism is associated with coronary artery disease, 367 consecutive patients who underwent percutaneous transluminal coronary angiography with stent implantation were genotyped. In patients (n=71) with diabetes, the A allele was associated with a smaller luminal diameter. In conclusion, a common functional polymorphism within the MMP-12 promoter influences coronary artery luminal dimensions in diabetic patients with manifest coronary artery disease.
Adult↗
Interleukin (IL)-10 inhibits IL-6 production in microglia by preventing activation of NF-kappaB.
The purpose of this study was to determine if interleukin (IL)-10 inhibits lipopolysaccharide (LPS)-induced IL-6 production in microglia by inhibiting activation of nuclear factor-kappaB (NF-kappaB). N13 microglia (a murine microglial cell line) and primary microglia from neonatal mice were cultured in the presence or absence of LPS and increasing amounts of murine IL-10 for 24 h. As predicted, LPS treatment increased supernatant IL-6 concentration in both N13 and primary microglia cultures. Pretreatment with IL-10, however, decreased LPS-induced IL-6 secretion in a dose-dependent manner in both culture systems. Likewise, ribonuclease protection assays showed that LPS increased steady-state IL-6 mRNA levels, but that pretreatment with IL-10 blocked the LPS-induced increase in IL-6 mRNA. Because NF-kappaB is the predominant transcription factor responsible for IL-6 transcription in response to inflammatory stimuli, it was hypothesized that IL-10 inhibited IL-6 production by preventing nuclear translocation of NF-kappaB. Consistent with this idea, LPS increased nuclear translocation of NF-kappaB as assessed by gel mobility shift assay. Supershift assays and immunocytochemical staining showed that both the p50 and p65 subunits of NF-kappaB translocated from the cytoplasm to the nucleus upon LPS stimulation. Pretreatment with IL-10, however, inhibited LPS-induced activation of NF-kappaB. Furthermore, inhibition of NF-kappaB activity with tosyl-Phe-chloromethlyketone (a serine protease inhibitor that prevents degradation of the NF-kappaB-IkappaB complex), completely blocked LPS-induced IL-6 production. These data suggest that IL-10 inhibited IL-6 production in microglia by decreasing the activity of NF-kappaB and, therefore, extend what little is known of the intricate relationship between anti-inflammatory and inflammatory cytokines in the central nervous system.
Animals↗
Nitric-oxide-dependent pial arteriolar dilation in the female rat: effects of chronic estrogen depletion and repletion.
In this study, we compared endothelial nitric oxide synthase (eNOS)-mediated cerebral vasodilating responses in intact female rats, chronically ovariectomized (OVX) rats, and OVX rats treated for 2 weeks with 17beta-estradiol (E(2)). Under anesthesia, using intravital microscopy and a closed cranial window system, pial arteriolar diameter changes were monitored during sequential cortical suffusions of an eNOS-dependent dilator [acetylcholine (ACh)] and a direct NO donor [S-nitrosoacetylpenicillamine (SNAP)]. In separate rats from the same groups, we compared eNOS and caveolin-1 (CAV-1) protein abundance in pial arterioles (via immunofluorescence analyses). In untreated and low-dose E(2)-treated (1.0 microg x kg(-1) x day(-1)) OVX rats, ACh-induced vasodilations were virtually absent. High-dose E(2) treatment (100 microg x kg(-1) x day(-1)) restored ACh-induced pial arteriolar dilations to levels seen in intact females. The vasodilations elicited by SNAP and ADO were unaffected by chronic estrogen changes, indicating no direct estrogen influence on vascular smooth muscle (VSM) reactivity. Pial arteriolar eNOS protein abundance was diminished by ovariectomy and restored by high-dose E(2) treatment. Pial arteriolar CAV-1 expression was higher in OVX versus intact and E(2)-treated OVX females. These results suggest that long-term changes in estrogen directly influence brain eNOS functional activity. The estrogen-related changes in eNOS-dependent vasodilating function appear to be related, in part, to a capacity for E(2) to increase eNOS protein expression and, in part, to an E(2)-associated diminution in endothelial CAV-1 expression.
Acetylcholine↗
Circular dichroism and resonance raman comparative studies of wild type cytochrome c and F82H mutant.
The UV-visible, circular dichroism (CD), and resonance Raman (RR) spectra of the wild type yeast iso-1-cytochrome c (WT) and its mutant F82H in which phenylalanine-82 (Phe-82) is substituted with His are measured and compared for oxidized and reduced forms. The CD spectra in the intrinsic and Soret spectral region, as well as RR spectra in high, middle, and low frequency regions, are discussed. From the analysis of the spectra, it is determined that in the oxidized F82H the two axial ligands to the heme iron are His-18 and His-82 whereas in the reduced form the sixth ligand switches from His-82 to Met-80 providing the coordination geometry similar to that of WT. Based on the spectroscopic data, the conclusion is that the porphyrin macrocycle is less distorted in the oxidized F82H compared to the oxidized WT. Similar distortions are present in the reduced form of the proteins. Frequency shifts of Raman bands, as well as the decrease of the alpha-helix content in the CD spectra, indicate more open conformation of the protein around the heme.
Amino Acid Substitution↗
Targeted gene repair in mammalian cells using chimeric oligonucleotides.
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Amino Acid Sequence↗
Rapid genotype analysis of the stromelysin gene 5A/6A polymorphism.
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Base Sequence↗
Ultrasound shear wave imaging for bone.
Recently, a computer-controlled scanning ultrasound (US) imaging system was developed in our laboratory. It includes a pair of broadband 48-MHz focusing copolymer transducers. The apertures of the transducers and their f-numbers were identically equal to 2 mm and 2.25, respectively. A specimen can be moved in a 10-microm increment in its plane and its normal direction can be rotated along the US propagation direction. It can be used to produce transmission-mode images of shear waves, as well as longitudinal waves for solid specimens. Shear waves in solids were generated by mode conversion. The results of the longitudinal and shear-wave US images for a piece of compact bovine bone obtained using this system are presented. Shear-wave images combined with longitudinal images can provide a more complete mechanical characterization of bone.
Animals↗
Rapid genotype analysis of the matrix metalloproteinase-1 gene 1G/2G polymorphism that is associated with risk of cancer.
A bi-allelic polymorphism in the promoter of the human matrix metalloproteinase-1 gene has been reported. It has been found to have a functional effect on the promoter strength and to be associated with risk of cancers. The polymorphism is due to an insertion/deletion of a guanine, and conventional methodologies for genotyping this polymorphism are time-consuming and expensive. A rapid genotyping method based on restriction endonuclease digestion is reported here.
Genotype↗