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Unified examination refers to the examination given by the university for medical students to graduate after they pass six curriculum examinations,including diagnostics,introduce of surgery,internal medicine,surgery,obstetrics and gynecology and pedology during the clinical study phases.It is a necessary process for cultivation of the professionals of the clinic medicine and has practice significance.Through the organization and management of the unified examination process,as well as analysis of examination result and feedback,education quality has been enhanced and educational reform promoted.
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Objective To compare the effects of ouabain and digoxin on both the systolic blood pressure and sodium pump α-subunit isoforms gene expression in the aortic smooth muscle of rats. Methods Normal SpragueDawley rats were injected with ouabain (20μg·kg-1 ·d-1 ,i. p),digoxin (32 μg·kg-1 ·d-1,i. p)and normal saline once a day, respectively, and indirect systolic blood pressure was recorded once a week. Six weeks later,all the rats were killed and sodium pump α1-,α2-,and α3-subunit mRNA levels were detected in the aortic smooth muscle with reverse transcription polymerase chain reaction(RT-PCR) method. Results The systolic blood pressure of rats infused with ouabain increased significantly at the end of week 6 [132. 6± 9. 0 mmHg (1 mmHg ＝ 0. 133 kPa)vs 115. 7±8.2mmHg, P ＜0. 01] ,while no difference of blood pressure was found between digoxin group and NS group (P＞0.05).The expression of sodium pump α-subunit isoforms in aortic smooth muscle was regulated by either ouabain or digoxin:both ouabain and digoxin increased α1- and α3-subunit expression, α2-subunit decreased in digoxin group but unchanged in ouabain group. Conclusion These results suggest that both ouabain and digoxin could regulate sodium pump α-subunit isoform expression, which might be related to the physiological roles of endogenous ouabain and might be responsible for the difference between the pharmacological and toxicological effects of ouabain and digoxin,including their effects on blood pressure.
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Objective To explore expression of endogenous ouabain(EO) in multiple adrenal tumors.Methods Thirty-one cases of adrenal tumors and 6 cases of healthy adrenal tissues were selected. The expression of EO in the adrenal tiss ue was detected with immunohistochemical streptavidin peroxadase conjugated(SP) method.Results Most of EO positive products were localized in cy toplasm of the zona reticularis of human adrenal cortex, and positive products s howed to be fine granular. There was no positive signal in the medulla. EO showe d on diffused positive in patients with pheochromocytoma accompanied high blood pressure[SBP:(165.22±7.61) mmHg, DBP:(105.52±4.26) mmHg], but there were neg ative in ones with normative blood pressure[SBP:(118.52±4.58) mmHg, DBP:(83±3.60) m mHg]. The expression of EO was positive in all adrenocortical hyperplasic, aden oma an d carcinoma, no matter its high or normative blood pressure. The degree of expre ssion of EO in adrenal tissues was related to the level of BP.Conclusion Expression of endogenous ouabain(EO) in health y adrenal tissue and adrenal tumors was a valuable morphological and pathophysio logical clue for the research on ouabain.
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Purpose The aim is to prepare ouabain polyclonal antibody F(ab)2 fragment and to estimate its molecular weight.Methods[ KG*2 [ WTBZ]Ouabain polyclonal antibody was obtained from immunized rabbits.The antibod y was digested with pepsin.The resulting products were analyzed and the molecular weig ht of F(ab)2 fragment was estimated by HPSEC.The immune activity was detec ted by ELISA.Results 100 mg of ouabain polyclonal antibody wa s dige sted by 2 mg of pepsin for 18 hours at pH 3.0 and active ouabain polyclonal anti body F(ab)2 framgment was obtained.Its molecular weight was 107 kD.Concl usion The active ouabain polyclonal antibody F(ab)2 fragment coul d be prepared by digesting its antibody with pepsin.
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Objective To investigate the gene expression of sodium pump α-subunit in aortic smooth muscle of 1-kidney-1-clip (1k1c) hypertensive rats. Methods 1k1c hypertensive rats were prepared by partially ligating the left renal artery and removing the right kidney. 4 weeks later, all the rats were killed and sodium pump α1- , α2-, and α3-subunit in aortic smooth muscles were detected with reverse transcription p ol ymerase chain reaction (RT-PCR) method and immunohistochemical assay at both mR NA and protein levels, respectively. Resulsts Changes in the expression of sodi um pump α-subunit gene were found in aortic smooth muscles of 1k1c hypertensiv e rats: α1-su bunit increased at both mRNA protein levels, while α2- and α3-subunits r emained without changes. Conclusions There were great changes in the gene expression of so dium pump α-subunit in aor tic smooth muscles of 1k1c hypertensive rats, which might be related to the development of hypertension in this hypertensive model.
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AIM: To explore the role of endogenous ouabain (EO) in the development of hypertension and its secretion character in 1k1c hypertensive rats(HR). METHODS: EO contents of plasma and tissues in 1k1c HR were detected by ELISA. The relationships between plasma and tissues ouabain and blood pressure were analyzed. RESULTS: EO contents of plasma, heart, kidney, adrenal gland, pituitary and hypothalamus in lklc HR were significantly higher than those of normal rats, especially in the adrenal gland and hypothalamus. EO contents of serum, kidney and hypothalamus were correlated with blood pressure. CONCLUSIONS: EO might play an important role in the development of hypertension in 1k1c hypertensive rats. Adrenal gland might be the major source of EO.
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Objective To compare the characteristics of endogenous ouabain(EO) secretion with the other adrenocortical hormones and determine the effects of angiotensin Ⅰ (Ang Ⅰ ), and adrenocorticotrophin(ACTH) on the secretion of EO. Methods EO was measured by radioimmunoassay from primary cultured bovine adrenocotical cells (BAC). Results ①Ouabain was determined in the media of cultured BAC. Both EO and aldosterone secretion were decreased from the outer to inner layer of the cultured adrenal cortex, and the responses to Ang Ⅰ and ACTH were higher than that in the mid layer (P ＜0. 05) and inner layer (P ＜0. 01). Cortisol secretion was activated by Ang Ⅱ or ACTH was significantly higher in the mid layer and in the inner layer than that in the outer layer. ②The time-course experiment showed that the gradually rising amounts of aldosterone and cortisol could be determined dur ing the continuous incubation to 48h with or without Ang Ⅰ or ACTH. However, EO did not increase continuously af ter 24h of incubation in the basal secreting situation and after 12h of incubation in the stimulating situation by Ang Ⅱ or ACTH. ③There were obvious drops in aldosterone and cortisol secretion from 3rd day during a 21 day-period cell culture, but the peak secretion of ouabain was in 7th day. Conclusion It suggests that the secretory mechanism might be different between EO and aldosterone or cortisol. Also, Ang Ⅱ and ACTH might be involved in the regulation of EO secretion.
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Objective To explore the relationship between endogenous ouabain and the pathogenesis of hyper- tension. Methods ① Sixteen Sprague-Dawley (SD) rats were selected and randomly divided into two groups, and then the rats were injected with ouabain in dosage of 20μg/kg. d and normal saline (NS), respectively. The indirect systolic blood pressure of all the rats were recorded once a week. ②) Twenty-five lk1c (one kidney and one clipped) hypertensive rats were established and injected randomly with anti-ouabain antibody, normal rabbit IgG and normal saline, respectively. The direct blood pressure of all the lklc hypertensive rats were recorded by cannulated carotid artery for 3h after administration. Results The systolic blood pressure of rats injected with ouabain began to increase 2 weeks after ouabain administration and increased significantly at 6 weeks compared with NS group (17.7±1.2)kPa vs .(15.4±1.1)kPa, P＜ 0. 01). Anti-ouabain antibody could significantly decrease the blood pressure of 1 k1c rats, while the normal rabbit IgG could not. ConclusionThe results of this study indicate that endogenous ouabain might be one of the pathogenetic factors of hypertension.
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AIM To select specific ouabain conjugated peptides (OCP) to decline ouabain level. METHODS Biopanning phage displayed 12 peptide library for OCP. The sequences of each selected peptide was determined and analyzed. The binding activity of synthetical OCP was identified by radioligand binding assay (RRA). RESULTS Three kinds of peptides were found out. Peptide A (12 peptide) was occupied in 0 67(8/12). The analysis of protein showed that there was no homogeneous protein in Genebank. The results of RRA showed that there were binding activity between synthetical OCP and 3H ouabain. Dissociation constant(K d) was 1 087 nmol?L -1 . Receptor density was 120 pmol?g -1 protein. IC 50 =1 46?10 -4 mol?L -1 . CONCLUSION It is important to obtain distinctive OCP, by which not only the protein sequence was analyzed, but also a valuable experimental data in detection of ouabain and therapy in hypertension also was offered.
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AIM: To investigate the regulation of sodium pump ?-subunit gene expression in the cortex of kidneys from one kidney one clip (1k1c) hypertensive rats. METHODS: 1k1c hypertensive rats were prepared by partially ligating the left renal artery and removing the right kidney. 4 weeks later, all the rats were killed, the levels of sodium pump ? 1-, ? 2-, and ? 3-subunit mRNA and protein in the cortex of kidney were detected with reverse transcription polymerase chain reaction (RT-PCR) method and immunohistochemical assay, respectively. RESULTS: The mRNA levels of sodium pump ? 1-subunit increased and ? 2- and ? 3-subunit were unchanged in the cortex of kidneys from 1k1c hypertensive rats compared with control rats, while no change has been found for all the three subunits gene expressions at protein level. CONCLUSION: There were some changes in the expression of sodium pump ?-subunit gene in the cortex of kidney of the 1k1c hypertensive rat, which might be related to the development of hypertension in this hypertensive model.