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1.
Electron. j. biotechnol ; 39: 74-81, may. 2019. tab, ilus, graf
Article in English | LILACS | ID: biblio-1052041

ABSTRACT

Background: CPEB is considered as an RNA-binding protein first identified in Xenopus oocytes. Although CPEB1 was involved in the growth of oocyte, its role in goat follicular granulosa cell has not been fully elucidated. To clarify the functions of this gene in goat follicular granulosa cells, CPEB1-overexpressing vector and interference vector were structured and transfected into follicular granulosa cells from Jiangsu native white goats of Nantong city, Jiangsu Province, China. The expression levels of differentiation-related genes including CDK1, Cyclin B1, and C-mos were determined 24 h after administration of CPEB1 by quantitative real-time polymerase chain reaction and Western blotting methods. Results: The expression levels of CDK1, Cyclin B1, and C-mos were significantly upregulated after overexpression and significantly downregulated after interference with CPEB1. Conclusions: The CPEB1 gene expression could affect the transcription of genes related to early cleavage divisions, which provided a reference for further research on its role in the growth and maturation of oocytes.


Subject(s)
Animals , Female , Oocytes , Transcription Factors/genetics , Goats/genetics , Transfection , Fertilization in Vitro , Gene Expression , Blotting, Western , Polymerase Chain Reaction/methods , RNA-Binding Proteins , Embryo Transfer , Livestock , Fluorescence , Granulosa Cells
2.
Electron. j. biotechnol ; 31: 17-23, Jan. 2018. tab, ilus, graf
Article in English | LILACS | ID: biblio-1022036

ABSTRACT

Background: CDC25 is a dual-specificity phosphatase that was first identified in the yeast Schizosaccharomyces pombe as a cell cycle-defective mutant. Although CDC25 is involved in the cell cycle of ovarian granulosa cells, the CDC25 signaling pathway has not been clarified fully. To explore the role of CDC25C in the cell cycle of goat ovarian granulosa cells, a CDC25C-overexpressing vector, pCMV-HA-CDC25C, was constructed and transfected into granulosa cells from adult and young white goats from Jiangsu Nantong. RT-PCR was used to measure CDC25C, CDK1, and WEE1 gene expression levels, and flow cytometry was used to distinguish ovarian granulosa cells in different phases of the cell cycle. Progesterone and estradiol levels in transfected ovarian granulosa cells were also measured. Results: In adult goat follicular granulosa cells transfected with pCMV-HA-CDC25C, CDC25C expression increased significantly, which greatly increased the relative gene expression levels of both CDK1 and WEE1. Additionally, progesterone and estradiol levels were increased in goat follicular granulosa cells overexpressing CDC25C. And the cell cycle results showed that transfection of pCMV-HA-CDC25C leads to a higher proportion of cells in S phase compared to the no vector-transfected groups. Conclusions: The results of this study indicated that the overexpression of CDC25C may increase the gene expression levels of both WEE1 and CDK1 in S phase and accelerate the transition of cells from G1 phase to S phase.


Subject(s)
Animals , Female , Goats , Cell Cycle/physiology , cdc25 Phosphatases/genetics , cdc25 Phosphatases/metabolism , Granulosa Cells/enzymology , Progesterone/analysis , Protein-Tyrosine Kinases/genetics , Transfection , Cell Cycle/genetics , Polymerase Chain Reaction/methods , Apoptosis , Cyclin-Dependent Kinases/genetics , Estradiol/analysis , Fertilization , Flow Cytometry , Fluorescence , Granulosa Cells/metabolism
3.
Chinese Medical Sciences Journal ; (4): 171-175, 2012.
Article in English | WPRIM | ID: wpr-243243

ABSTRACT

<p><b>OBJECTIVE</b>To construct the zinc finger protein-activating transcription factor (ZFP-ATF) plasmid and evaluate its efficacy in inducing vascular endothelial growth factor (VEGF) expression in EY.HY926 endothelial cells.</p><p><b>METHODS</b>Firstly, we constructed the ZFP-ATF plasmid, then testified the quantity of VEGF protein in EY.HY926 endothelial cells after transfected with ZFP-ATP plasmid by Western blot, finally, we used the RT-PCR to testify whether the ZFP-ATF can stimulate expression of VEGF splice variants.</p><p><b>RESULTS</b>The ZFP-ATF DNA sequences were located the multiclone sites of PVAX1 vector between the site of BamH1and Xhol.Western blot result showed VEGF expression in EY.HY926 endothelial cells transfected with ZFP-ATF plasmid was significantly higher than that in cells transfected with VEGF165 (19.95±3.95 vs.12.15±1.55 μg÷μL, P<0.01).RT-PCR result showed VEGF-A mRNA expression level induced by ZFP-ATF was high than that induced by VEGF165.</p><p><b>CONCLUSION</b>ZFP-ATF can up-regulate the VEGF-A expression in comparison with VEGF165, which might have beneficial effects in angiogenesis process.</p>


Subject(s)
Humans , Activating Transcription Factors , Physiology , Amino Acid Sequence , Base Sequence , Cells, Cultured , Endothelial Cells , Metabolism , Molecular Sequence Data , Neovascularization, Physiologic , Plasmids , Up-Regulation , Vascular Endothelial Growth Factor A , Genetics , Zinc Fingers , Physiology
4.
Acta Academiae Medicinae Sinicae ; (6): 142-145, 2011.
Article in English | WPRIM | ID: wpr-341442

ABSTRACT

<p><b>OBJECTIVE</b>To evaluate the role of multidetector row spiral computed tomography in the diagnosis of popliteal artery entrapment syndrome (PAES).</p><p><b>METHOD</b>Ten patients (13 limbs) were diagnosed as PAES through open surgery in Peking Union Medical College Hospital from January 2006 to December 2009, among whom three (30.0%) had both limbs involved.</p><p><b>RESULTS</b>Eleven limbs (84.6%) had occlusions in popliteal artery at admission. While only 15.4% of the patients were diagnosed as PAES by positional stress test, Doppler ultrasound, and arterial angiography, the disease was confirmed in all 10 patients by spiral computed tomography. As shown by inter-operative exploration, the PAES types included type 1 (n=2, 15.4%), 2(n=5, 38.5%),3 (n=4, 30.8%), and 5 (n=2, 15.4%). Two limbs underwent popliteal artery release only, 8 limbs underwent politeal artery release, thromboendarterectomy, and angioplasty with a venous or prosthetic patch, and 3 patients underwent catheter-based thrombolysis and popliteal artery release subsequently. During the follow-up (mean: 22 months; range: 4-33 months), the primary patency rate was 92.3% (12/13), the total patency rate was 100% (13/13), and the limb salvage rate was 100%.</p><p><b>CONCLUSION</b>Positional stress test, Doppler ultrasound, and arterial angiography play limited roles in the diagnosis of PAES with popliteal artery occlusion, while multidetector row spiral computed tomography is much helpful in diagnosing and typing PAES.</p>


Subject(s)
Adolescent , Adult , Female , Humans , Male , Middle Aged , Young Adult , Angiography , Methods , Follow-Up Studies , Multidetector Computed Tomography , Peripheral Vascular Diseases , Diagnostic Imaging , Popliteal Artery , Diagnostic Imaging , Retrospective Studies
5.
Chinese Medical Sciences Journal ; (4): 241-245, 2011.
Article in English | WPRIM | ID: wpr-299377

ABSTRACT

<p><b>OBJECTIVE</b>To evaluate the efficacy of surgical therapy for carotid body tumors.</p><p><b>METHODS</b>A retrospective analysis was conducted, covering the diagnosis, surgical procedure, postoperative complications, and prognosis of 120 cases of carotid body tumors in Peking Union Medical College Hospital from 1949 to May, 2011.</p><p><b>RESULTS</b>Surgical excision was successfully performed in 111 cases with 117 tumors. In all those cases, 50 underwent simple tumor resection, 42 underwent resection of tumors and ligation of the external carotid arteries, 7 underwent co-resection of tumors and common carotid arteries, internal carotid arteries, as well as external arteries without vascular reconstruction, and the other 12 cases experienced tumor resection and vascular reconstruction as internal carotid arteries were involved. After operation, 3 cases developed cerebral infarction, 30 cases showed cranial nerve palsy, including 15 cases of hypoglossal nerve damage, 10 cases of vagus paralysis, and 5 cases of Horner's syndrome.</p><p><b>CONCLUSION</b>It is essential to make a proper surgical strategy, which can reduce postoperative complications.</p>


Subject(s)
Adolescent , Adult , Aged , Female , Humans , Male , Middle Aged , Carotid Body Tumor , General Surgery , Follow-Up Studies , Postoperative Complications , Epidemiology , Retrospective Studies
6.
Chinese Medical Sciences Journal ; (4): 116-119, 2003.
Article in English | WPRIM | ID: wpr-321402

ABSTRACT

<p><b>OBJECTIVE</b>To describe our surgical experience on rupture of abdominal aortic aneurysm.</p><p><b>METHODS</b>Two cases of ruptured aortic aneurysms with severe complication were analysed. Aorta reconstruction procedures were performed using bifurcated e-PTFE grafts during emergency operation. Diagnosis, preoperative resuscitation, emergency surgical intervention, and postoperative complications of these patients were summarized and discussed.</p><p><b>RESULTS</b>Rupture of aortic aneurysm in both patients presented as a huge retroperitoneum haematoma by computed tomography scan. They were successfully saved by prompt body fluid compensation, emergency procedure, intraoperative resuscitation, and postoperative intensive care.</p><p><b>CONCLUSIONS</b>Correct diagnosis, prompt surgical management, immediate intraoperative proximal aorta clamping during procedure, and effective management of postoperative complications were the key points to successful treatment of ruptured aortic aneurysm.</p>


Subject(s)
Aged , Humans , Male , Aortic Aneurysm, Abdominal , Diagnosis , General Surgery , Aortic Rupture , Diagnosis , General Surgery , Follow-Up Studies , Postoperative Complications , Therapeutics
7.
Chinese Medical Sciences Journal ; (4): 220-224, 2002.
Article in English | WPRIM | ID: wpr-244887

ABSTRACT

<p><b>OBJECTIVE</b>To evaluate the possibility and efficiency of nanoparticle as a new vector in specific gene transference.</p><p><b>METHODS</b>Nanoparticle-DNA complex was prepared with Poly-dl-lactic-co-glycolic acid (PLGA) bearing anti-sense monocyte chemotactic protein-1 (A-MCP-1), a specific expression gene, and the package efficiency, release progress in vitro, and the size of the complex were determined. The possibility of the new vector was evaluated with genomic DNA PCR by transferring gene into cultured smooth muscle cells (SMC), cationic lipids as a control. For study in vivo, jugular vein-to-artery bypass grafting procedures were performed on 20 New Zealand white rabbits, of which 6 grafts were transferred with nanoparticle-A-MCP-1 (200 microg), 6 with A-MCP-1 (200 microg) by cationic liposome, 4 with LNCX plasmid, and 4 as control. Fourteen days after the grafts were harvested, the expression of A-MCP-1 and its effect on MCP-1 in vein grafts were detected by dot blot, and the morphologic evaluation of grafts was performed.</p><p><b>RESULTS</b>The package efficiency of the nanoparticle-DNA complex was 0.9%, release progress in vitro lasted 2 weeks, and the size ranged from 150 to 300 nm. SMC genomic DNA PCR showed that A-MCP-1 gene could be successfully transfected into cells by nanoparticle. The study in vivo indicated that A-MCP-1 mRNA was expressed in both local gene delivery groups, nanoparticle and liposome, meanwhile, MCP-1 expression in vein grafts was significantly inhibited and neointimal hyperplasia was notably reduced.</p><p><b>CONCLUSION</b>Nanoparticle can act as a vector to transfect specific gene.</p>


Subject(s)
Animals , Rabbits , Chemokine CCL2 , Genetics , DNA, Antisense , Genetics , Drug Carriers , Gene Expression , Genetic Therapy , Genetic Vectors , Lactic Acid , Nanotechnology , Particle Size , Polyglycolic Acid , Polymers , Transfection
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