Biomedical subjects

Dao-Wen Wang

Publications and source records attributed to Dao-Wen Wang.

10 recordsLinked to original sources

PubMed2005

[Influences of recombinant adeno-associated virus-mediated Decorin gene transfection on cell cycle and apoptosis of SiHa cells].

BACKGROUND & OBJECTIVE: Decorin (DCN), a multi-efficiency factor, has many important functions, such as influencing fibril stability, and interacting with extracellular matrix molecules to influence cell adhesion. This study was to investigate influences of DCN on cell cycle and apoptosis of SiHa cells via recombinant adeno-associated virus-mediated DCN gene (rAAV.DCN) transfection. METHODS: Based on the cDNA sequence of DCN reported by GenBank, DCN segment with open reading frame was cloned into plasmid UF(1) (UF(1)-DCN) after DNA sequencing. rAAV.DCN, containing high titer of DCN, was produced by transfecting into 293 cells with calcium phosphate-DNA sedimentation method, and transfected into SiHa cells, rAAV.LacZ-infected cells and non-infected cells were used as controls. Expression of DCN was determined by Western blot. Cell cycle and apoptosis were analyzed by flow cytometry (FCM). RESULTS: High titer of rAAV .DCN was produced successfully. After transfected with rAAV.DCN, SiHa cells effectively expressed DCN. The percentage of G(1) phase in rAAV.DCN-transfected cells was significantly higher than that in control cells [(72.3+/-2.3)% vs. (59.5+/-2.5)%, P<0.05], while the percentages of S phase, and G(2) phase in rAAV.DCN-transfected cells were significantly lower than those in control cells [(11.9+/-1.9)% vs. (17.8+/-0.8)%, and (13.4+/-1.1)% vs. (21.5+/-0.9)%, P<0.05]. No change of apoptosis was detected in rAAV.DCN-transfected cells. CONCLUSION: Cell cycle of SiHa cells may be arrested by high expression of DCN.

Apoptosis↗

PubMed2005

[Wheat transformation by electroporation with ring electrode].

Electroporation has been used effectively to deliver DNA into the tissue of intact wheat immature embryos. The physical parameters of electroporation were 770 V/cm field strength, 800 microF capacitor and 100 microg/mL of plasmid DNA, containing bar and GUS gene. The electroporation was carried out by a portable and permanent ring electrode that can be fitted into the wells of 24-well plates. The samples were pulsed three times. Integration of the introduced genes into the genome of transgenic wheat plants was shown by PCR and Southern blot analysis. Transformation frequency was 7.5%, that is higher than that of the 4.2% frequency by microprojectile bombardment.

DNA, Plant↗

PubMed2005

[Effects of CD151 on migration of human tongue squamous carcinoma cell line Tca8113].

BACKGROUND & OBJECTIVE: Over-expression of CD151 gene in tumor tissues may be associated with metastasis and poor prognosis of cancer, but the mechanism is unknown. This study was designed to determine the effects of CD151 gene on migration of human tongue squamous carcinoma cell line Tca8113. METHODS: Functional fragment of CD151 gene was amplified by reverse transcription-polymerase chain reaction (RT-PCR), and inserted into plasmid adeno-associated virus (pAAV) vector in sense direction and antisense direction to construct rAAV-CD151 and rAAV-antiCD151, the virus titers were determined by dot blot. Two weeks after transfection, protein level of CD151 in Tca8113 cells was detected by Western blot, the effect of CD151 on migration of Tca8113 cells was detected by Transwell chambers. RESULTS: Titer of rAAV-CD151 was 2x10(11) pfu/ml, titer of rAAV-antiCD151 was 1x10(11) pfu/ml. Transfection of rAAV-CD151 increased expression of CD151 in Tca8113 cells by 108% of control cells, while transfection of rAAV-antiCD151 decreased expression of CD151 in Tca8113 cells by 79% of control cells. Migrated cells count was significantly higher in rAAV-CD151-transfected cells (93.6+/-11.6), and lower in rAAV-antiCD151-transfected cells (24.0+/-4.4) than in untransfected and rAAV-GFP-transfected cells (53.0+/-6.6 and 46.0+/-7.0, P<0.05). CONCLUSIONS: It is an important molecular mechanism of tumor metastasis that over-expression of CD151 promotes migration of tumor cells. rAAV-antiCD151, a novel therapeutic tool, can specifically reduce the expression of CD151, and inhibit migration of tumor cells.

Antigens, CD↗

PubMed2005

Inducing rye 1R chromosome structural changes in common wheat cv. Chinese spring by the gametocidal chromosome 2C of Aegilops cylindrica.

To generate 1 R deletion and translocation lines, we introduced a 2C chromosome,which was derived from Aegilops cylindrica and was known to have a gametocidal function when added monosomically into common wheat cv. Chinese Spring (CS) and its derivative, into a wheat-rye 1R chromosome disomic addition line (CS-1R"). When the individuals with chromosome constitution 21" + 1R" + 2C' (2n = 45) were selfed, the 1R chromosome structural changes were found to be induced with high frequency (24.1%) among the progenies. By using C-banding and GISH analysis, we analyzed 1R structural changes in 46 F3 individuals, which came from 23 F2 plants. The rearranged 1R chromosomes could be characterized in about 85% of the F3 individuals. This included telosome 1RL (39.1%), iso-chromosome 1 RL (2.2%), whole arm translocation involving 1RL (32.6%), telosome 1RS (4.3%), iso-chromosome 1RS (4.3%), and 1R deletion mutant with break point in the long arm (2.2%). The mutant 1R lines obtained in this study will potentially be useful in mapping the chromosome locations of agronomically important genes located in 1R. This study also demonstrated that molecular markers might be used to identify wheat chromosome arm involved in translocation with 1R.

Chromosome Aberrations↗

PubMed2004

Expression of cyclooxygenase-2 in human esophageal squamous cell carcinomas.

AIM: To determine whether cyclooxygenase-2 (COX-2) was expressed in human esophageal squamous cell carcinoma. METHODS: Quantitative reverse transcription-polymerase chain reaction (RT-PCR), western blotting, immunohistoc-hemistry and immunofluorescence were used to assess the expression level of COX-2 in esophageal tissue. RESULTS: COX-2 mRNA levels were increased by >80-fold in esophageal squamous cell carcinoma when compared to adjacent noncancerous tissue. COX-2 protein was present in 21 of 30 cases of esophageal squamous cell carcinoma tissues, but was undetectable in noncancerous tissue. Immunohistochemistry was performed to directly show expression of COX-2 in tumor tissue. CONCLUSION: These results suggest that COX-2 may be an important factor for esophageal cancer and inhibition of COX-2 may be helpful for prevention and possibly treatment of this cancer.

Adult↗

PubMed2004

[Association between mutation of phospholamban gene and dilated cardiomyopathy].

Phospholamban (PLB) is a prominent regulator of myocardial contractility and a reversible inhibitor of the cardiac sarcoplasmic reticulum Ca2+ ATPase activity. In normal cardiac muscles, phospholamban can be phosphorylated at distinct sites by various protein kinases and release its inhibition to sarcoplasmic reticulum Ca2+ ATPase. The studies of pedigrees have shown dilated cardiomyopathy (DCM) is related with mutation of PLB. The aim of present study is to investigate the relationship between mutation of PLB gene and DCM. Sixty patients with idiotic DCM were enrolled in present study. The clinical data were collected, including clinical symptoms, ECG and echocardiography. Peripheral blood samples of all these subjects were collected to extract genome DNA. The fragments of PLB gene were amplified by PCR and PCR fragment sequencing was performed to study weather mutation of phospholamban gene exists. phospholamban gene did not show any mutation in these patients. Most Chinese DCM patients may not be related with mutation of PLB gene.

Adolescent↗

PubMed2003

[Adeno-associated virus vector carrying human minidystrophin gene SMCKA3999 effectively ameliorates dystrophic pathology in mdx model mice].

OBJECTIVE: Duchenne muscular dystrophy (DMD) is the most common and letal genetic skeletal muscle disorder, caused by recessive mutations in the dystrophin gene and no treatment is available. The present paper is aimed to study if recombinant adeno-associated virus vector (rAAV) mediated dystrophin minigene SMCKA3999 could effectively ameliorates dystrophic pathology in mdx model mice. METHODS: We used dystrophin minigene SMCKA3999 to construct rAAVSMCKA3999. When injected into the skeletal muscle of mdx mice (DMD model), we adopted methods of immunofluorescent (IF) staining, Evans Blue and electromicroscopy to observe if rAAVSMCKA3999 could effectively ameliorates dystrophic pathology in mdx model mice. RESULTS: rAAVSMCK3999 resulted in efficient and stable expression and restoration of the missing dystrophin onto the plasma membrane. rAAVSMCKA3999 can also protect myofiber membrane integrity. For the first time we prove that rAAVSMCKA3999 can improve ultrastructure changes of DMD. CONCLUSION: We have demonstrated the effectiveness of rAAVSMCKA3999 in correcting pathology changes of skeletal muscle, which offer a promising avenue for DMD gene therapy.

Animals↗

PubMed2003

Serine/threonine kinase activity in the putative histidine kinase-like ethylene receptor NTHK1 from tobacco.

A histidine kinase-based signaling system has been proposed to function in ethylene signal transduction pathway of plants and one ethylene receptor has been found to possess His kinase activity. Here we demonstrate that a His kinase-like ethylene receptor homologue NTHK1 from tobacco has serine/threonine (Ser/Thr) kinase activity, but no His kinase activity. Evidence obtained by analyzing acid/base stability, phosphoamino acid and substrate specificity of the phosphorylated kinase domain, supports this conclusion. In addition, mutation of the presumptive phosphorylation site His (H378) to Gln did not affect the kinase activity whereas deletion of the ATP-binding domain eliminated it, indicating that the conserved His (H378) is not required for the kinase activity and this activity is intrinsic to the NTHK1-KD. Moreover, confocal analysis of NTHK1 expression in insect cells and plant cells suggested the plasma membrane localization of the NTHK1 protein. Thus, NTHK1 may represent a distinct Ser/Thr kinase-type ethylene receptor and function in an alternative mechanism for ethylene signal transduction.

Adenosine Triphosphate↗

PubMed2002

Effects of the angiotensin receptor antagonist losartan on the response of the left main coronary artery to cold pressor test in patients with essential hypertension as assessed by echocardiography.

BACKGROUND: Angiotensin II can impair endothelial function, which is mediated by the angiotensin II type 1 receptor subtype. OBJECTIVE: To test the hypothesis that treatment with the angiotensin II type 1 receptor antagonist losartan would restore the normal dilation of the left main coronary artery to the cold pressor test in patients with essential hypertension, as assessed by echocardiography. PATIENTS AND METHODS: The study population included 24 patients with mild to moderate essential hypertension and 24 matched, healthy subjects. The cold pressor test-induced and nitroglycerin-induced changes in the left main coronary artery diameter were measured by echocardiography at the end of the washout period and after 12 weeks of losartan administration. RESULTS: The percentage change in the left main coronary artery diameter induced by the cold pressor test in patients with hypertension (-4.3 8.7%) was significantly lower than that in control subjects (10.5 3.9%, P<0.0001). The percentage change in the left main coronary artery diameter induced by the cold pressor test in patients with essential hypertension after losartan treatment (13.7 8.0%) was significantly higher than that before losartan treatment (-4.3 8.7%, P<0.0001). The percentage change in the left main coronary artery diameter induced by the cold pressor test after losartan treatment was not significantly different between the 14 patients with hypertension who had satisfactory control of blood pressure (14.1 8.8%) and the 10 patients with hypertension who did not have satisfactory control of blood pressure (13.1 7.0, P=0.8). Losartan treatment in patients with essential hypertension did not modify the percentage change in the left main coronary artery diameter caused by sublingual administration of nitroglycerin (23.2 14.4% versus 27.2 13.5, P=0.2). CONCLUSIONS: The present study demonstrates that treatment with the angiotensin II type 1 receptor antagonist losartan normalized the response of the left main coronary artery to the cold pressor test in patients with mild to moderate essential hypertension, and that this effect is not dependent on the reduction of blood pressure.

Adult↗

PubMed2002

Arachidonate CYP hydroxylases of kidney contribute to formation of hypertension and maintenance of blood pressure.

AIM: To investigate the relationship of kidney-specific expression of cytochrome P-450 (CYP) 4A1 and the blood pressure. METHODS: The specific sense and antisense CYP4A1 cDNA was administered respectively with the help of eukaryotic expression vector pcDNA3.1 to the Sprague-Dawley (SD) rats via sublingual vein (2 mg/kg). The systolic blood pressure of rats was assessed by the tail- cuff method, and the relative tissue expression of CYP4A1 was analyzed by Western blot and Northern blot at RNA and protein levels in the brain, heart, lung, liver, and kidney of control, sense, and anti-sense CYP4A1 cDNA-treated rats. RESULTS: Two weeks after the injection of the sense and antisense CYP4A1 cDNA recombinants respectively, the mean systolic pressure of the transgenic rats increased by 1.8 kPa +/- 0.3 kPa (13.2 mmHg +/- 2.5 mmHg) or decreased by 1.7 kPa +/- 0.3 kPa (13.0 mmHg +/- 2.2 mmHg) compared with control. At the levels of transcription and translation, the Northern and Western blots all demonstrated that CYP4A1 preferentially overexpressed in the kidney. CONCLUSION: The administration of sense and antisense CYP4A1 cDNA induced hypertension and hypotension, respectively, which indicated that renal arachidonate hydroxylase contributed to the formation of hypertension and maintenance of blood pressure in normotensive rats.

Animals↗