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Objective To clone the R1-MYB transcription factor participated in the anthocyanidin metabolism, and to analyze by bioinformatics analysis. Different expression of different varieties, different organs of the same species and salt stress conditions in Lycium were analyzed. To clone the full-length cDNA encoding R1-MYB, to perform bioinformatic analysis, and to study its expression in different cultivators and different developmental stage and in response to NaCl stress in Lycium ruthenicum and L. barbarum. Methods The full-length cDNA encoding R1-MYB was cloned using homology-based cloning and rapid amplification of cDNA ends (RACE) technique in L. ruthenicum, and the homologous gene was obtained by transcriptome in L. barbarum. The bioinformatics analysis was carried out by using Prot, Param, Smart, PSORT, and SOPMA methods. And the phylogenetic tree was constructed based on software MEGA5.0. Gene expression analysis was done by method of Real-time PCR. Results We the MYB transcription factor in L. ruthenicum was cloned and named as LrMYB1R1 (GenBank accession number KY568981), and LbMYB1R1 (GenBank accession number KY568982) in L. barbarum. Bioinformatics analysis showed that the length of LrMYB1R1 was 1 496 bp and the CDS was 927 bp. The coding products contained 308 amino acids, the molecular weight of the protein was 33 400 and 33 490, the theoretical isoelectric point was 7.80 and 7.78, belonging to the R1-MYB transcription factor, and the encoded protein is predicted to be located in the nucleus. The results of phylogenetic tree analysis showed that LrMYB1R1 and LbMYB1R1 were highly similar to MYB1R1-like protein in Solanum lycopersicum, Solanum tuberosum, and Nicotiana tabacum. Real-time PCR analysis showed that LrMYB1R1 had higher expression level in leaves and young fruits in L. ruthenicum, followed by stems, young leaves, flowers, purple fruits and black fruits, only slightly expressed in roots. In addition, the relative expression levels of LrMYB1R1 decreased in response to salt stress. Conclusion The study of R1 MYB transcription factor has been enriched, which has laid the foundation for the subsequent research on gene function and for the high-yielding anthocyanin by genetic engineering method in L. ruthenicum.
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Objective To evaluate treatment strategies for fungal endocarditis after heart prosthetic valve surgery. Methods Two cases of severe fungal infection after heart prosthetic valves surgery were analyzed retrospectively, related literatures were reviewed.Results Two patients had fungal endocarditis after surgery,the valve function was affected,patients were hospitalized repeatedly after surgery.In case 1 ,fever occurred 45 days after cardiac sur-gery and patient was returned to the hospital for re-examination,emergency mitral valve replacement was performed under cardiopulmonary bypass,the postoperative vegetation culture suggested Aspergillus flavus.In case 2,the aortic wall vegetation was removed 5 months after heart surgery under cardiopulmonary bypass,pathology of post-operative vegetation suggested mucor.Two patients were promptly removed infection foci through surgery and trea-ted with standard antifungal agents,patient with Aspergillus infection died after rescue,and patient with mucor in-fection was cured,the latter was more powerful in antifungal therapy.Conclusion Prevention is the key to fungal endocarditis after heart prosthetic valve surgery,treatment should be prompt and effective,antifungal agents should be given in sufficient dose and course.
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<p><b>OBJECTIVE</b>To study the clinicopathologic features and expression of epidermal growth factor receptor (EGFR) and vascular endothelial growth factor (VEGF) in adrenocortical tumors.</p><p><b>METHODS</b>Forty-two cases of adrenocortical tumors operated at the Beijing Union Medical College Hospital during the period from July, 2001 to July, 2010 were retrospectively reviewed. Immunohistochemical study for EGFR and VEGF was carried out. The clinical information and follow-up data were analyzed.</p><p><b>RESULTS</b>The cases included 21 adrenocortical carcinomas (ACC) and 21 adrenocortical adenomas (ACA). Nine patients suffered from primary aldosterone syndrome, including 8 cases with ACA and 1 case with ACC. The average tumor size, tumor weight, and duration between disease onset and diagnosis in the 21 cases of ACC were 11.7 cm, 542 g and 8.5 months, respectively. This was in contrast to 3 cm, 9.8 g and 45.6 months, respectively in cases of ACA. Histologically, the WEISS score in all the 21 cases of ACA was ≤ 2 (average = 0.9). None of the ACC cases had score less than 4 (average = 6.6). The presence of sinus invasion correlated with tumor metastasis (P < 0.01). Immunohistochemical study showed that EGFR was expressed in 61.9% of ACC patients (13/21), whereas EGFR staining was mostly negative in ACA (except for weak staining in 5 cases and moderate staining in 1 case). The difference of EGFR expression between ACC and ACA was statistically significant (P = 0.030). On the other hand, the positive rate of VEGF in ACC was 71.4% (15/21), including 28.6% (6/21) with strong expression and 28.6% (6/21) with moderate expression. In contrast, the expression rate of VEGF in ACA was 30.0% (7/21), including 14.3% (3/21) with moderate expression. The difference of VEGF expression between ACC and ACA was statistically significant (P = 0.013). There was correlation between VEGF expression and venous invasion (P = 0.028). The average duration of survival in patients with ACC was shorter than that in ACA. The tumor weight in ACC also correlated with prognosis.</p><p><b>CONCLUSIONS</b>Tumor size, weight and presence of endocrine symptoms may help in the differential diagnosis between ACC and ACA. A WEISS score of ≥ 3 highly suggests ACC. The presence of sinus invasion is associated with metastasis. EGFR or VEGF expression may also be important in differentiating ACC from ACA.</p>
Subject(s)
Adolescent , Adult , Aged , Child , Child, Preschool , Female , Humans , Male , Middle Aged , Young Adult , Adrenal Cortex Neoplasms , Metabolism , Pathology , General Surgery , Adrenocortical Adenoma , Metabolism , Pathology , General Surgery , Adrenocortical Carcinoma , Metabolism , Pathology , General Surgery , Diagnosis, Differential , Follow-Up Studies , ErbB Receptors , Metabolism , Retrospective Studies , Survival Rate , Tumor Burden , Vascular Endothelial Growth Factor A , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To investigate the effect of OX40/OX40L interaction on the nuclear factor of activated T cells c1 (NFATc1) in ApoE-/- mice.</p><p><b>METHODS</b>Lymphocytes were prepared from mouse spleens after Collar-treated Surgery, then incubated with a range of agonistic anti-OX40 mAbs and inhibitory anti-OX40L mAb to stimulate or inhibit OX40-OX40L interaction in vitro. The expression of NFATc1 mRNA and protein in lymphocytes of ApoE-/- mice was measured by Real Time PCR and flow cytometry, respectively.</p><p><b>RESULTS</b>(1) After stimulating OX40-OX40L signal pathway, the expression of NFATc1 mRNA and protein in leukocytes of ApoE-/- mice was significantly increased, with maximal effect occurring at 20 µg/ml anti-OX40 mAb-stimulated, and peaked at 24 h at any concentration (P < 0.01). (2) Anti-OX40L mAb significantly suppressed the expression of NFATc1 in leukocytes of ApoE-/- mice, with maximal effect occurring at 20 µg/ml anti-OX40L mAb, and peaked at 24 h (P < 0.001).</p><p><b>CONCLUSIONS</b>OX40-OX40L interaction can regulate the mRNA and protein expressions of NFATc1 in lymphocytes of ApoE-/- mice.</p>