Yuan et al press oleic acid-mediated membrane permeability transition (MPT) of mitochondria through acceleration of beta-oxida- tion. The opening of MPT has been proved as a key event in cell apoptosis or necrosis after IR injury [12, 13]. It has been suggested that L-carnitine would prevent apoptosis and maintain functions of cell. The purpose of this experiment is to explore the effects of L-carnitine on the rats suffering from the small intestinal IR injury through detecting translo- cation of EmGFP labelled E. coli from the mesenteric lymph nodes, liver, spleen and portal vein blood. Additionally, the small intestinal morphometric study was performed and the levels of serum cytokines (TNF-α, IL-1β, IL-6 and IL-10) were investigated. Materials and Methods Animals A total of thirty adult SD rats were obtained from the Ex- perimental Research Center of Zhengzhou University, and their weight is in the range of 280 - 320 g. All of the rats were maintained under standard conditions with controlled temperature (18 ± 4 °C), 12 hours light and 12 hours dark- ness cycle, and feeding with standard food and water. The maintenance of the experimental rats was administrated by licensed operators and these rats were given humane treat- ment. This study was approved by the Ethical Committee of Kunming Medical University. Labeling of Escherichia coli The Escherichia coli DH5α (donated from PHD Cao, the De- partment of Micro-organism of Kunming Medical Univer- J Clin Med Res • 2011;3(2):78-84 Figure 1. The E. coli with GFP were detected with ﬂuores- cence microscope. The cells displayed rod shape and erupt- ed ﬂuorescent light. sity) was selected to label with EmGFP. The brief procedures are as follows. 1. Establishment of competent bacteria using CaCl2 method. 2. Transformation of plasmid DNA (pRSET- EmGFP bought from the Invitrogen Company) into E. coli using the heat shock method. A mixture of the competent bacteria and the plasmid DNA were placed in iced bath for 30 minutes and put at 42 °C for 2 minutes (heat shock), and then placed back into the iced bath. After transformation, the bacteria were cultured on Luria-Bertani plate with ampicil- lin (100 mg/L) at 37 °C for 12 hours. Four colonies were inoculated into liquid media (200 ml) and grown at 37 °C with agitation for 16 hours in order to ensure growth of iso- lated colonies. The E. coli DH5α with transformed plasmid containing EmGFP were detected by using ﬂuorescence mi- Table 1. The Chiu’s Score Classiﬁcation of Small Intestinal Injury Classiﬁcation Pathological change Level 0 Level 1 Level 2 Level 3 Level 4 Level 5 Mucosa without changes. Well-constituted villosities, no cellular lysis or inﬂammatory process, although there is formation of Grunhagen’s sub-epithelial space. Presence of cellular lysis, formation of Grunhagen’s sub-epithelial space and increased spacing among the villosities. Destruction of the free villosities section, presence of dilated capillaries and inﬂamed cells. Structural destruction of the villosities, only traces of some villosities, formed by inﬂamed cells and necrotic material, with hemorrhage and basal glandular ulceration. Destruction of all the mucosa, no glandular structure can be seen, only the amorphous material laying on the sub-mucosa tissue. Articles © The authors | Journal compilation © J Clin Med Res and Elmer Press™ | www.jocmr.org 79