Latest Issue
Volume 25 ， Issue 2 ， 2010
- Abstract：Objective To investigate the effect of microRNA-205 reduction by antagomirs on adhesion ability of normal human corneal epithelial keratinocytes (NHCEKs). Methods Antagomir-205, complementary and inhibitory to microRNA-205, was used to suppress endogenous microRNA-205 in NHCEKs. The adhesion ability of treated NHCEKs was then assessed by cell adhesion assay. Immunoblot and immunohistochemistry were conducted to determine the level of two focal adhesion-related proteins, focal adhesion kinase (FAK) and paxillin (Pax). Phalloidin staining was performed to measure the level of filamentous actin in antagomir-treated NHCEKs. Results Antagomir-205 markedly reduced the level of microRNA-205 in NHCEKs and significantly enhanced adhesion ability of NHCEKs (P<0.01). Further protein analysis validated that inhibition of microRNA-205 increased the number of phosphorylated FAK and phosphorylated Pax, and decreased filamentous actin. Conclusion Our findings suggest that microRNA-205 has down-regulating effect on cell motility in NHCEKs.Keywords：corneal epithelium;cell signaling;microRNA4|5305|0Updated：2024-04-10
- Abstract：Objective To investigate the regulatory mechanisms of acetylated p53 in the expression of microtubule-associated protein-2 (MAP2) in neuronal differentiation of P19 cells induced by all-trans retinoic acid (RA). Methods Neuronal differentiation of P19 cells was initiated with 4-day RA treatment. Immunofluorescence, real-time reverse transcription-polymerase chain reaction (RT-PCR) assay, and map2 promoter driven luciferase assay were performed to detect the expression and relative promoter activity of MAP2 in those RA-treated cells. Real-time PCR-based chromatin immunoprecipitation assay (ChIP) was carried out to reveal the specific recruitment of acetylated p53 onto its binding sites on map2 promoter. Results The expression of MAP2 was markedly increased in RA-induced P19 cells. The map2 mRNA increased 34-fold after 4 days of RA treatment and 730-fold 2 days after the treatment, compared with the cells without RA treatment (control). p53 was recruited to the promoter of map2 gene in acetylated form and thereby enhanced its promoter activity. p300/CBP associated factor (PCAF) was found induced in RA-treated cells and enriched in the nucleus, which might contribute to the acetylation of p53 in the regulation of map2 gene. Conclusions Acetylated p53 may participate in regulating the expression of map2 in RA-induced differentiation of P19 cells. PCAF is possibly involved in this process by mediating the acetylation of p53.Keywords：p53;p300/CBP associated factor;microtubule-associated protein-2;retinoic acid;neuronal differentiation4|3140|0Updated：2024-04-10
- Abstract：Objective To investigate the role of acyl-coenzyme A: cholesterol acyltransferase inhibitor (ACATI) in apoptosis induced by lipids and whether lipids-induced apoptosis is accompanied by increase of free cholesterol in endoplasmic reticulum (ER), in order to further understand the mechanism of lipids-induced apoptosis in advanced atherosclerosis. Methods Human vascular smooth muscle cells (VSMCs) and phorbol 12-myristate 13-acetate (PMA) differentiated THP-1 macrophages were used. Tritiated thymidine incorporation was applied to detect cell proliferation. Cytotoxicity was assessed by lactate dehydrogenase (LDH) release. 4',6-diamidino-2-phenylindole (DAPI) staining, caspase-3, -7 assay, and Annexin-V/propidium iodide (PI) staining were used to detect apoptosis. High performance liquid chromatography was used in intracellular free cholesterol and cholesterol ester assay. ER free cholesterol was quantified. Results Different lipids had different effects on proliferation and cytotoxicity of VSMCs. 25-hydroxycholesterol (25OHC) had biphasic effects on the proliferation of VSMCs. At low concentration, it stimulated cell proliferation, but turned to proliferation inhibition as concentration reached 15 ug/mL. 25OHC and acetylated low density lipoprotein (AcLDL) could respectively induce apoptosis in human VSMCs and PMA differentiated THP-1 macrophages, which was aggravated by ACATI, accompanied by increase of intracellular free cholesterol content. There was also an increase of cholesterol content in ER with AcLDL-induced apoptosis in THP-1 macrophages. Conclusions Lipids could induce apoptosis, accompanied by increase of intracellular free cholesterol content, which could be augmented by ACATI, suggesting that insults resulting in ER free cholesterol rise might be the initiator of apoptosis.Keywords：apoptosis;atherosclerosis;acyl-coenzyme A: cholesterol acyltransferase6|4006|0Updated：2024-04-10
- Abstract：Objective To study the association of DTNBP1 gene with some symptom factors of schizophrenia.Methods A total of 285 unrelated schizophrenic individuals were recruited from December 2004 to January 2009 for genetic analysis, and their symptom factors were assessed based on the Positive and Negative Syndrome Scale (PANSS). The quantitative trait test was performed by the UNPHASED program (version 3.0.12) to investigate the association between scored positive and negative symptoms and the single nucleotide polymorphisms (SNPs) in DTNBP1 gene.Results The quantitative trait test showed allelic association of rs909706 with the excitement symptom of schizophrenia (P<0.05, adjusted by 10 000 permutations), while the genotype C/G of rs2619539 with a negative symptom, lack of spontaneity and flow of conversation (P<0.05, adjusted by 10 000 permutations).Conclusion DTNBP1 variations are possibly associated with some symptoms of schizophrenia, which could partly explain the relationship between the susceptibility gene DTNBP1 and that disease.Keywords：schizophrenia;DTNBP1 gene;positive and negative symptoms;quantitative trait test4|4310|0Updated：2024-04-10
- Abstract：Objective To assess the expression level of D-Tyr-tRNATyr deacylase (DTD) in SAMP8 mice and speculate the function of DTD in disorders associated with Alzheimer's disease (AD). Methods Altogether 12 SAMP8 mice and 12 SAMR1 mice were used in this study. Semi-quantita- tive reverse transcription-polymerase chain reaction (RT-PCR) and Western blot were performed to detect the mRNA and protein levels of DTD in the mice. Purified DTD protein was injected into lateral ventricle to investigate the function of DTD in SAMP mice. The behavior of the mice was tested by using a Step-through Test System. Results Both mRNA and protein levels of DTD were found to be significantly lower in SAMP8 mice compared with those in SAMR1 mice (P<0.05). In vivo injection of DTD protein did not lead to an obvious change in behavior of SAM mice. Conclusions DTD might function in the process of AD-associated pathology and could possibly participate in physiology process in a long-term manner to orchestrate with other regulators in order to maintain the balance of organism.Keywords：D-Tyr-tRNATyr deacylase;D-amino acid;SAMP8 mice;neurodegenerative disease4|3257|0Updated：2024-04-10
- Abstract：Objective To develop an alternative method for assessment of gene delivery systems in vivo. Methods Mouse primary spleen lymphocytes were genetically modified in vitro by a retroviral vector harboring a Gaussia luciferase (Gluc) expression cassette. After implantation of these cells into recipient mice, the expression of Gluc was detected in whole blood or plasma collected. Results As little as 10 uL whole blood drawn from the recipient mice could guarantee prompt reading of Gluc activity with a luminometer. And the reading was found in good correlation with the number of genetically modified spleen lymphocytes implanted to the mice. Conclusions Gluc may be useful as an in vivo reporter for gene therapy researches, and Gluc blood assay could provide an alternative method for assessment of gene delivery systems in vivo.Keywords：gene delivery systems;reporter;Gaussia luciferase4|3263|0Updated：2024-04-10
- Abstract：Objective To investigate the molecular mechanism of nectin-like molecule 1 (NECL1) inhibiting the migration and invasion of U251 glioma cells. Methods We infected U251 glioma cells with adeno-nectin-like molecule 1 (Ad-NECL1) or empty adenovirus (Ad). Transwell and wound healing assays were performed to observe the migration of U251 cells incubated with the cell supernatant from Ad-NECL1 or Ad infected U251 cells. DNA microarray was applied to screen the gene expression profile after the restoration of NECL1 in U251 glioma cell lines. The differential expression of osteopontin (OPN), a gene related to migration and invasion, was further analyzed with semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR), Western blot, and immunohistochemistry. Results The restoration of NECL1 inhibited migration of U251 cells significantly (P<0.05). Altogether 195 genes were found differentially expressed by microarray, in which 175 were up-regulated and 20 down-regulated, including 9 extracellular matrix proteins involved in the migration of cells. Both mRNA and protein expressions of OPN, the most markedly reduced extracellular matrix protein, were found decreased in U251 cells after restoration of NECL1. Immunohistochemical assay also detected an increase of OPN in glioma tissues, related with the progressing of malignant grade. Conclusion A link might exist between NECL1 and the extracellular matrix protein OPN in inhibiting the migration and invasion of U251 glioma cells.Keywords：nectin-like molecule 1;glioma cell line;extracellular matrix protein;osteopontin4|3259|0Updated：2024-04-10
- Abstract：Objective To study the association between the epidermal growth factor (EGF) gene and intelligence in patients with major depression. Methods Intelligence measurement using Wechsler Adult Intelligence Scale (WAIS) was perfor- med on 120 unrelated patients with major depression and 46 control subjects. Blood was collected from all subjects for extraction of genomic DNA. Four single nucleotide polymorphisms (SNPs) in the EGF gene were genotyped using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI- TOF-MS). Results Mean scores of both score lang and score task, two subtests in WAIS, differed significantly between major depression patients and controls (P<0.0001). Quantitative trait analysis showed that the genetype of rs2250724 was closely associated with score lang and score task in major depression patients. The associations were still significant after 10 000 permutations. Conclusions Although preliminary, our results provide evidence for association between the EGF gene and intelligence in patients with major depression. Genetic variation in the EGF gene may increase the susceptibility of major depression.Keywords：epidermal growth factor;intelligence;major depression6|3521|0Updated：2024-04-10
- Abstract：Objective To investigate the feasibility of analyzing acylcarnitine in dry filter-paper blood spots by liquid chromatography-tandem mass spectrometry (LC-MS/MS) which could be applied to detect inborn errors of metabolism in neonates. Methods We obtained filter-paper blood from 26 dead infants from a neonatal intensive care unit (NICU) between October 1, 2008 and September 30, 2009. Acylcarnitine and amino acid profiles were obtained with LC-MS/MS. Four infants underwent routine autopsy. The postmortem blood specimens were compared with newborn blood specimens, and with specimens obtained from older infants with metabolic disorders. Results Of all the 26 patients, 5 (19.2%) were diagnosed as having different kinds of diseases: 3 with methylmalonic acidemia (the concentration of C3, and the ratio of C3/C16, C3/C2 increased), 1 with maple syrup urine disease (the concentration of leucine and isoleucine increased), and 1 with isovaleric aci- demia (the concentration of C5 increased). Conclusions Postmortem metabolic test can explain infant deaths and provide estimates of deaths attributable to inborn errors of metabolism in NICU. LC-MS/MS is suitable for analysis of postmortem specimens and can be considered for routine application in NICU autopsy.Keywords：tandem mass spectrometry;inborn errors of metabolism;neonatal intensive care unit;autopsy4|3990|0Updated：2024-04-10
- Abstract：Objective To assess recent trends and prognostic features in the treatment of pure mucinous carcinoma (PMC) of the breast. Methods Fifty-six patients diagnosed with PMC of the breast in our hospital from December 1982 to June 2008 were included. We evaluated the general information and tumor characteristics of the patients, examined the relationship between these factors and prognosis. Fisher's exact test was applied to analyze tumor characteristics. Results The mean age of the patients was 53.7 years. The majority of the patients presented with early stage disease. Tumor size was found not a significant prognostic factor in this study. Mean follow-up period was 39 months and no breast cancer-related deaths were identified in the patient cohort. Conclusions PMC of the breast has a favorable prognosis. Tumor size does not appear to significantly impact survival.Keywords：breast cancer;pure mucinous breast carcinoma;lymph node status4|3623|0Updated：2024-04-10
- Abstract：Caveolin-2, a protein about 20 kD, is a major component of the inner surface of caveolae, small invaginations of the plasma membrane. Similar with caveolin-1 and caveolin-3, it serves as a protein marker of caveolae. Caveolin-1 and -2 are located next to each other at 7q31.1 on human chromosome, the proteins encoded are co-localized and form a stable hetero-oligomeric complex, distributing similarly in tissue and cultured cells. Caveolin-3 is located on different chromosomes but confirmed to interact with caveolin-2. Caveolin-2 is similar to caveolin-1 in many respects but differs from the latter in functional domains, especially in G-protein binding domain and caveolin scaffolding domain. The mRNAs of both caveolin-1 and caveolin-2 are most abundantly expressed in white adipose tissue and are induced during differentiation of 3T3-L1 cells to adipocytes. Caveolin-2-deficinet mice demonstrate clear pulmonary defects, with little or no change in caveolin-1 expression and caveolae formation, suggesting that caveolin-2 plays a selective role in lung functions. Caveolin-2 is also involved in lipid metabolism and human cancers.Keywords：caveolin-2;lung;lipid metabolism;cancer4|3754|0Updated：2024-04-10
- Abstract：PAGET’S disease, also called osteitis deformans, is a metabolic bone disorder. It is characterized by increased bone resorption and the compensatory formation of new bones. The? increased bone conversion and remodeling lead to the incrustation of woven bones and lamellar bones and finally result in the expansion, loosening, and excessive vascularization of the affected bones, rendering them susceptible to deformity and fracture.1 Paget’s disease occurs much more commonly in Anglo-Saxons than in Asians and Africans. Only a few cases have been reported in China, and the condition in those cases involved the pelvis and tibia.2 A patient with Paget’s disease was admitted into our hospital in June 2009 and was treated with surgery for secondary thoracic spinal stenosis. Given the rarity of Paget’s disease, we expected to contribute to the study in this field by reporting the case with spinal involvement requiring surgical treatment, which has rarely been documented so far.Keywords：Paget's disease;thoracic spinal stenosis;surgical decompression4|3329|0Updated：2024-04-10
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