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| 1 | Degradation of intestinal mRNA:A matter of treatment显示文摘AIM: To characterize the influence of location, species and treatment upon RNA degradation in tissue samples from the gastrointestinal tract. METHODS: The intestinal samples were stored in different medium for different times under varyingconditions: different species(human and rat), varying temperature(storage on crushed ice or room temperature), time point of dissection of the submucous-mucous layer from the smooth muscle(before or after storage), different rinsing methods(rinsing with Medium, PBS, RNALater or without rinsing at all) and different regions of the gut(proximal and distal small intestine, caecum, colon and rectum). The total RNA from different parts of the gut(rat: proximal and distal small intestine, caecum, colon and rectum, human: colon and rectum) and individual gut layers(muscle and submucosal/mucosal) was extracted. The quality of the RNA was assessed by micro capillary electrophoresis. The RNA quality was expressed by the RNA integrity number which is calculated from the relative height and area of the 18 S and 28 S RNA peaks. From rat distal small intestine q PCR was performed for neuronal and glial markers.RESULTS: RNA obtained from smooth muscle tissue is much longer stable than those from submucosal/mucosal tissue. At RT muscle RNA degrades after one day, on ice it is stable at least three days. Cleaning and separation of gut layers before storage and use of RNALater, maintains the stability of muscle RNA at RT for much longer periods. Different parts of the gut show varying degradation periods. RNA obtained from the submucosal/mucosal layer always showed a much worse amplification rate than RNA from muscle tissue. In general RNA harvested from rat tissue, either smooth muscle layer or submucosal/mucosal layer is much longer stable than RNA from human gut tissue, and RNA obtained from smooth muscle tissue shows an increased stability compared to RNA from submucosal/mucosal tissue. At RT muscle RNA degrades after one day, while the stability on ice lasts at least three days. Cleaning and separation of gut layers before storage and use of RNALater, maintains the stability of muscle RNA at RT for much longer periods. Different parts of the gut show varying degradation periods. The RNA from muscle and submucosal/mucosal tissue of the proximal small intestine degrades much faster than the RNA of distal small intestine, caecum or colonwith rectum. RNA obtained from the submucosal/mucosal layer always showed a much more reduced amplification rate than RNA from muscle tissue [β-Tubulin Ⅲ for muscle quantification cycle(Cp): 22.07 ± 0.25, for β-Tubulin Ⅲ submucosal/mucosal Cp: 27.42 ± 0.19].CONCLUSION: Degradation of intestinal m RNA depends on preparation and storage conditions of the tissue. Cooling, rinsing and separating of intestinal tissue reduce the degradation of m RNA. | Sabine Heumüller-Klug Carsten Sticht Karin Kaiser Elvira Wink Cornelia Hagl Lucas Wessel Karl-Herbert Schafer | 2015 | World Journal of Gastroenterology2015，21，12： | 2 |
| 2 | Replacement of ascending aorta with aortic valve reinplantation : midterm results 显示文摘 | Harringer WP Pething K Hagl C | 1999 | Eur J Cadiothorac Surg1999，15，6： | 1 |
| 3 | Prevalence of diabetes and impaired glucose tolerance in patients treated with conventional depot neuroleptie medications 显示文摘 | Hagl S Joelsson L Mjorndal T | 1998 | J Clin Psychiatry1998，59，6： | 1 |
| 4 | Die Aortendissektion – Wann operieren, wann endoluminal therapieren?显示文摘 | G.M. Richter J.R. Allenberg H. Schumacher J. Hansmann C. Vahl S. Hagl | 2001 | Der Radiologe2001，，8： | 1 |
| 5 | Technical advances of pul-monary thromboendarterectomy for chronic thromboembolic pulmo-nary hypertension显示文摘 | Hagl C Khaladj N Peters T | 2003 | Eur J Cardiothorac Surg2003，23，5： | 1 |
| 6 | Angiotensin induces inflammatory activation of human vascular smooth muscle cells显示文摘 | Kranzhofer R Schmidt J Pferffer CA Hagl S Lippy P Kubler W | 1999 | Arterioscler Thromb Vasc Biol1999，19，： | 1 |
| 7 | Aortic root surgery in Marian syndrome : Comparison of aortic valve-sparing reimplantation versus composite grafting 显示文摘 | Karck M Kallenbach K Hagl C | 2004 | J Thorac Cardiovasc Surg2004，127，： | 1 |
| 8 | The safety of moderate hypothermic lower body circulatory arrest with selective cerebral perfusion:a propensity score analysis显示文摘 | Kamiya H Hagl C Kropivnitskaya I | 2007 | J Thorac Cardiovasc Surg2007，133，2： | 1 |
| 9 | Neurologic outcome after ascending aorta-aortic arch operations:effect of brain protection technique in highrisk patients显示文摘 | Hagl C Ergin MA Galla JD | 2001 | J Thorac Cardiovasc Surg2001，121，6： | 1 |
| 10 | Significance of the subvalvular apparatus for left ventricular dimensions and systolic function:experimental replacement of the mitralvalve显示文摘 | GAMS E HAGL S SCHAD H | 1991 | Thorac Cardiovas Surg1991，39，1： | 1 |
| 11 | Smooth muscle proteins from Hirschsprung disease facilitates stem cell differentiation显示文摘 | Hagl CT Heumller S Klotz M | 2012 | Pediatr Surg Int2012，28，2： | 1 |
| 12 | The safety of moderate hypothermic lower body circulatory arrest with selective cerebral perfusion: A propensity score analysis显示文摘 | Hiroyuki Kamiya Christian Hagl Irina Kropivnitskaya | 2007 | The Journal of Thoracic and Cardiovascular Surgery2007，133，50： | 1 |
| 13 | The microenvironment in the Hirschsprung : s disease gut supports myenteric plexus growth显示文摘 | Hagl CI Rauch U Klotz M | 2012 | Int J Colorectal Dis2012，27，6： | 1 |
| 14 | Results of valve sparing aortic root reconstruction in 158 consecutive patients 显示文摘 | Kallenbach K Hagl C Walles T | 2002 | Ann Thorac Surg2002，74，： | 1 |
| 15 | Respirafory depression in adult patients with intravenous patient-controlled analgesia 显示文摘 | Hagle ME Lehr VT Brubakken K | 2004 | Orthop Nuts2004，23，1： | 1 |
| 16 | Valve-sparing aortic root stabilization in acute type a aortic dissection 显示文摘 | Shrestha M Khaladj N Hagl C | 2009 | Asian Cardiovasc Thorac Ann2009，17，1： | 1 |
| 17 | Poly (ADP-ribose) polymerase activation in the reperfused myocardium 显示文摘 | SzaboG Liaudet L Hagl S | 2004 | Cardiovasc Res2004，61，3： | 1 |
| 18 | Blocking the hedgehog pathway inhibits hepatoblastoma growth 显示文摘 | Eichenmuller M Gruner I Hagl B | 2009 | Hepatology2009，49，2： | 1 |
| 19 | BlocKing the hedgehog pathway inhibits hepatoblastoma growth显示文摘 | Eichemntlller M Gruner I Hagl B | 2009 | Hepatology2009，49，2： | 1 |
| 20 | Respiratory depression inadult patients with intravenous patient-controlled analgesia显示文摘 | Hagle ME Lehr VT Brubakken K | 2004 | Orthop Nurs2004，23，1： | 1 |
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