临床医学英语论文.doc

Abstract: objective to study of TNF alpha Ghrelin human brain microvascular endothelial cells induced (HBMEC) mononuclear cells chemotaxis protein 1 (MCP-1) influence and p38MAPK role. Methods HBMEC training, TNF alpha group to join different concentration TNF alpha; Ghrelin pretreatment section first join Ghrelin pretreatment 1 h, then add the TNF alpha. The RT-PCR method to detect MCP-1 mRNA level, immune imprinting method to detect p38 phosphorylation (p-p38) expression. The results of the TNF alpha stimulation, MCP-1 mRNA express obvious enhancement, and the concentration of a certain dose-response relationship (P 0. 05) in the cytoplasm and P-p38 protein expression also significantly increased (P 0. 05). Ghrelin pretreatment can reduce MCP-1 mRNA cytoplasm and the p-p38 protein expression (p 0. 05). Conclusion Ghrelin may inhibit p-p38 pathways inhibit TNF alpha mediated HBMEC MCP-1 mRNA expressions.Keywords: mononuclear cells chemotaxis protein 1; The human brain microvascular endothelial cells; GhrelinIn recent years, and the relationship between Ghrelin inflammation is paid attention to, but for inflammation, Ghrelin in different tissues function is not the same, need to make clear further 1 5. April 2008 to July 2009, we studied Ghrelin TNF alpha induction of the human brain microvascular endothelial cells (HBMEC) mononuclear cells chemotaxis protein 1 (MCP-1) phosphorylation mRNA and p38 (p-p38) the influence of the expression, this paper discusses Ghre-Lin in the possible anti-inflammatory mechanism vessel, as Ghrelin applied to clinical diabetes cerebrovascular disease and provided the reference.1 materials and methods1. The human brain microvascular endothelial cell lines 1 materials HBMEC Scien cell bought in the United States, rhTNF alpha Sigma company bought in, TaKaRaRNA PCR Kit (AMV) Ver. 3. 0 bought in dalian treasure biological engineering Co., LTD, p-p38MAPK mice monoclonal antibodies against human bought in Santa Cruz company, other drugs for the sold analysis pure.1. 2 method1. 2. 1 cells culture and group HBMEC cells use tire bovine serum containing 5% of 37 and 5% in DMEM CO2Humidity under cultivation, and pancreatic enzyme digestion ChuanDai, the fusion status for the experiment. With 5 x 105 / hole vaccination to 6 orifice plate, to 90% fusion, serum-free culture 24 h, PBS cleaning 2 times, join condition culture medium: TNF alpha (0. 1, 1, 10 ng/ml) or Ghrelin (1, 10, 100 ng/ml) effect 24 h.Ghrelin pretreatment section first join Ghrelin (10 ng/ml) pretreatment 1 h, then add the TNF alpha, and the concentration of 1. 0 ng/ml, training 24 h, abandon medium to join TRIzol reagent, cracking cells, extraction RNA.1. 2. 2 MCP-1 mRNA express detection using total RNA extraction reagent TRIzol, cells extracted total RNA, uv spectrophotometer measurement sample spectrophotometry, A260 / A280 nm should be in 1: 2. 0. Primer according to GenBank middleman MCP-1 cDNA sequence design, MCP-1 upstream: 5 GCTCGCTCAGCCAGATGCAAT-3 , downstream: 5 TGGGTTGTGGAGTGAGTGTTC-3 , the expansion of section for 257 bp; Beta actin on tour: 5 AAATCGTGCGTGACAT-TAA-3 , downstream: 5 CTCGTCATACTCCTGCTTG-5, amplification clips for 473 bp. RT-PCR concrete operation according to illustrate kit. Drug synthesis cDNA reaction conditions: 30 10 min, 42 30 min, 99 10 min, 5 5 min. Synthesis of making cD with primer for corresponding amplification. Expansion conditions for 94 the degeneration 2 min, with 94 degeneration 1 min, 57 annealing 1 min. 72 extensions 1 min, a total of 30 circulation, 72 extensions 10 min. PCR products by 1. 5% agarose gel electrophoresis gel after imaging system analysis. Light density scanner light density detection, and with MCP-1 mRNA and beta actin mRNA said the ratio of the relative value of making the mR. Take control of the relative expression for 1, each of the relative amount compared with the expression with multiple of said.1. 2. 3 p-p38 protein expression detection with 100 ng/ml Ghrelin pretreatment cell 1 h joined after 10 ng/ml of TNF alpha, cultivate 30 min respectively, collect cells, extraction protein, immune imprinting method to detect p-p38, beta actin expression. With p-p38 and beta actin density of the ratio of the said purpose the relative content of protein, the relative expression for the control group of 1, each of the relative amount compared with the expression with multiple of said.1. 2. 4 statistical methods using SPSS 13. 0 software comparison between groups, measurement data with 珋 x s said, compares the two two LSD method. With P than 0. 05 for statistically significant.2 the results2. 1 MCP-1 mRNA without the express HBMEC cells stimulate MCP-1 mRNA expression level is low, TNF alpha stimulation, MCP-1 express obvious enhancement, the concentration of a certain dose-response relationship (P 0. 05), see figure 1. Ghrelin alone to MCP-1 role to express the impact is not big, but Ghrelin pretreatment can reduce TNF alpha induction of MCP-1 mRNA expression (P 0. 05), see figure 2.2. 2 p-p38 Ghrelin protein expression of TNF alpha induction of cytoplasmic p-p38 protein expression influence the results shown in figure 3. The figure 3 shows, normally, HBMEC cytoplasmic express certain p-p38 protein that TNF alpha stimulate 30 min, the cytoplasmic p-p38 protein significantly increased, and Ghrelin separate function group p-p38 protein expression the normal group is less; Ghrelin pretreatment group in the cytoplasm p-p38 protein is TNF alpha group reduce (p 0. 05).3 discussThere is evidence that the metabolic syndrome in express MCP-1, TNF alpha channel have special role. To determine whether the endothelial Ghrelin cerebrovascular modulate inflammatory factor MCP-1 secretion, we tested the Ghrelin of MCP-1 mRNA influence. The results showed that TNF alpha obviously increase the human brain microvascular endothelial cell lines HBMECMCP-1 mRNA expression, its may be obese patients with insulin resistance to get one of the causes of the cerebral arteriosclerosis. Ghrelin can restrain the foundation and TNF alpha induction of MCP-1 mRNA expression, prove its can inhibit endothelial inflammatory factor expression of cerebral function, this is Ghre-Lin independent of adjusting glucolipid metabolism outside of the role, is also the main points of the study. PetCO2 monitoring. For patients with lung disease, lung disease will delay the potential CO2 to eliminate, more should be extended breath support after the time. This kind of patients PetCO2 mild increase and was not accurate in reflecting artery blood high CO2 and respiratory acidosis degree, the determination of the Pet-CO2 value and the blood PaCO2 correlation is not good. Postoperative premature dial to the spontaneous breathing easy appear hypercapnia and acidosis, should first selection arterial blood gas monitoring. The CO2 gasless laparoscopic led to internal pressure within the chest and pressure, blood vessels active substances such as catecholamine and vasopressin release increase, hypercapnia evoked the sympathetic nervous tension increased. Can cause significant hemodynamic changes. We in the study also found that patients and stop after gasless laparoscopic 15, pp 30 min SBP before a clinical increases, and HR and DBP over time points, DBP SBP and HR is a slightly elevated before pp, but no clinical significance. May the choice of case and case and intraoperative strict management relevant, patients with no serious disease, maintain enough depth of anesthesia, ensure analgesia and perfect control gasless laparoscopic relatively low pressure are perioperative blood flow dynamics of the important factors less jerky. This also with our previous observation results are basically the same. Elderly patients for clinical operation cycle reflect mainly displayed in the SBP on the rise of the concrete reason is not very clear, pending further study.In conclusion, we through to over 60 patients after laparoscopic study found that, between PetCO2 and PaCO2 are good for the correlation. PetCO2 can be accurate guide intraoperative breathing management, after laparoscopic gasless laparoscopic in elderly patients. The influence of the SBP obviously, perioperative should pay attention to monitoring and handle in time.Reference：1：Soubani AO Noninvasive monitoring of oxygen and carbon dioxideJ Am J Emerg Med，2001，19( 2) : 141-1462：杨明华，林智平，叶允荣 不同潮气量机械通气下肺癌根治术病人动脉血二氧化碳分压与呼气末二氧化碳分压的关系J 中华麻醉学杂志，2006，26( 1) : 86