1.Protein kinases involved in the endothelial nitric oxide synthase activation by beta-adrenoceptors stimulation in human umbilical vein endothelial cells
Chinese Journal of Tissue Engineering Research 2008;12(33):6589-6592
BACKGROUND: Beta-adrenergic activation can enhance signal-transduction pathway of endothelial nitric oxide (NO). Vascular endothelial NO production in response to β -adrenergic activation is important in the normal control of vessel tone by the sympatheadrenal system. Previous trials have demonstrated that endothelial nitric oxide synthase (eNOS) activated by isoprenaline, β -adrenergic agonist, may be correlated with phosphorylation of eNOS. OBJECTIVE: To investigate the possible protein kinase involved in the signal transduction of the eNOS activation by β -adrenoceptor (β AR) stimulation in human vascular endothelium. DESIGN: Comparative observation. SETTING: Department of Geriatrics, First Affiliated Hospital of Zhengzhou University. MATERIALS: The experiment was performed at the Open Key Clinic Medical Experimental Laboratory of University of Henan Province between September 2006 and June 2007. Fresh umbilical cords were obtained following delivery of healthy babies to healthy normotensive mothers, either by vaginal delivery or by elective Caesarean section. The written informed consent was obtained from all donors. Approval was granted by the Affiliated Hospital of Zhengzhou University. Isoprenaline, H-89, Wortmannin, [3H]-L-arginine were from Sigma, USA. METHODS: Blank control group, H-89 group (protein kinase inhibitor), Wortmannin group (phosphatidylinositol 3-kinase inhibitor) and H-89+Wortmannin group were set up, and isoprenaline subgroup and blank subgroup were subdivided with 3 tubes in each group. Human umbilical vein endothelial cells (HUVECs) were cultured with H-89 and Wortmannin, separately for 10 minutes, then agonist isoprenaline or vehicle was added and the incubation continued for another 30 minutes, eNOS activity was determined by the conversion of [3H]-L-arginine to [3H]-L-citrulline. The results were corrected by protein concentration in the corresponding cellular extracts. MAIN OUTCOME MEASURES: Changes in eNOS activity of each group. RESULTS: ①Basal [3H]-L-citrulline production was (3 085.62±272.72) mBq per 1 μ g protein, lsoprenaline increased [3H]-L-citrulline formation by (30.72 ± 3.91 )% compared with the control group (P < 0.001 ). H-89 or Wortmannin alone did not alter eNOS activity [(0.44± 1.00)%, (0.36±1.47)%, P > 0.051. ②The increase of eNOS activity by isoprenaline wassignificantly inhibited by pretreatment of H-89 and Wortmannin [(4.73±2.19)%, (17.35±3.72)%, P < 0.001]. ③The inhibition of protein kinase and phosphatidylinositol 3-kinase to isoprenaline-induced increase in eNOS activity [(4.92± 0.99)%] was similar to protein kinase alone, but not in an additive effect (P > 0.05). CONCLUSION: Protein kinase and phosphatidylinositol 3-kinase are involved in the signal transduction of β- adrenoceptors mediated eNOS activation.
2.Consistency of gene amplification and its expression at protein level of human epidermal growth factor receptor 2 in gastric carcinoma
Xinyan ZHANG ; Juanjuan FU ; Wenjing PU ; Zhenzhu SUN
Chinese Journal of Digestion 2013;33(10):680-683
Objective To investigate the consistency of gene amplification and its expression at protein level of human epidermal growth factor receptor 2 (HER2) in gastric carcinoma.Methods From 2010 to 2012,120 gastric cancer specimens of patients with gastric cancer were collected,of which 100 were surgical specimens and 20 were specimens from biopsy under gastroscope.The protein expression of HER2 in 120 specimens was detected by immunohistochemistry (IHC).According to the results of IHC,the positive parts of HER2 expression of IHC slices were developed into tissue microarrays for fluorescence in situ hybridization (FISH) to test the gene amplification of HER2.The different parts with different color intensity of focal (+ + +) (≤ 10% tumor cell strongly staining) specimens detected by IHC detection were compared with the results of FISH.Kappa test was performed for statistical analysis.Results Among 120 gastric cancer specimens,the results of IHC indicated that 77 specimens were positive with different staining intensity including 16 strong positive (+++),six focal positive (+++),37 moderate positive (++) and 18 weak positive (+).The positive rate of HER2 protein expression detected by IHC was 18.3% (22/120).The results of FISH showed 41 specimens were positive and the rate of gene amplification was 34.2%.Among which,21 were moderate positive (++) detected by IHC,15 were strong positive (+ + +) and five were focal positive (+ ++).The positive rate of HER2 was 35.0% (42/120) with IHC and FISH combined detection.The consistent rate of IHC and FISH was 91.6 % (76/83).Kappa coefficient was 0.960 (P<0.01).In five positive specimens detected by FISH and which were focal positive (+ + +) by IHC,the different parts with different color intensity were compared with the results of FISH and gene amplification was found in all specimens.Conclusion Tissue microarray technology is consistent with IHC in HER2 detection in gastric cancer specimens and could help to improve the detection rate.
3.Effects of"5E" rehabilitation scheme on transitional care in post-discharge patients with cerebral hemorrhage
Fen XUE ; Juanjuan LU ; Pu YANG ; Gaofeng RAO ; Mei MA ; Zhe WANG
Chinese Journal of Modern Nursing 2017;23(18):2353-2356
Objective To explore the effects of "5E" rehabilitation scheme on transitional care in post-discharge patients with cerebral hemorrhage.Methods A total of 148 inpatients with cerebral hemorrhage in a hospital the author was in were selected in observation group from June 2014 to May 2016 along with implementing transitional care with the "5E" rehabilitation scheme. A total of 144 inpatients with cerebral hemorrhage in a hospital the author was in were selected in control group from June 2012 to May 2014 with routinely transitional care. Patients' motor function, self efficacy and quality of life was compared between two groups after intervention.Results After implementing nursing intervention with the "5E" rehabilitation scheme, the scores of motor function in upper and lower limbs of patients in control group were all higher than those in control group with significant differences (t=3.216, 2.921;P<0.01). The changing degree of 10 dimensions in sport scale, disease information, getting help, communication with physicians, general disease control, doing housework, social entertainments, symptom management, rapid breath management, depression control of observation group before and after nursing intervention were greater than those of control group with significant differences (t=4.862, 5.641, 4.848, 4.252, 3.875, 3.793, 3.765, 3.314, 3.324, 3.832;P<0.01). The improvements in the score of physiological function, mental function, social function, ability of health self-awareness and the total score of quality of life of patients in observation group were higher than those in control group with significant differences (t=5.985, 4.784, 4.732, 4.556, 3.873;P<0.01).Conclusions The "5E" rehabilitation scheme can improve the motor function, self-management skills and quality of capability.