Objective To investigate the effect of mesenteric lymphatic duct liagtion and glutamine enteral nutrition on intestine and distant organs in intestinal ischemia/reperfusion injury. Methods Forty male SD rats undergoing gastrostomy were randomly assigned into 5 groups (n=8): sham operation group, normal enteral nutrition group, normal enteral nutrition+lymphatic duct ligation group, glutamine group and glutamine+lymphatic duct ligation group. Sham operation group only received laparotomy after 7 days of full diet, the other four groups were subjected to 60 min of intestinal ischemia after 7 days of enteral nutrition, and the two lymphatic duct ligation groups were plus mesenteric lymphatic duct ligation. The original nutrition continued 3 days after reperfusion. Intestinal permeability was detected on day 1 before reperfusion, day 1 and 3 after reperfusion. Intestinal morphology was observed, endotoxin, D-lactate and diamine oxidase levels in serum, and apoptotic index in lung tissue were detected on day 3 after reperfusion. Results The intestinal permeability in each group was significantly increased on day 1 after reperfusion (Plt;0.05), and which in normal enteral nutrition+lymphatic duct ligation group and glutamine+lymphatic duct ligation group were significantly decreased on day 3 after reperfusion (Plt;0.05). The mucosal thickness and villus height of ileum and mucosal thickness of jejunium in glutamine+lymphatic duct ligation group were significantly higher than those in other groups (Plt;0.05), and villus height of ileum in glutamine group was higher than that in normal enteral nutrition group (Plt;0.05); those morphology indexes in normal enteral nutrition+lymphatic duct ligation group were higher than those in normal enteral nutrition group, but there was no statistical signification (Pgt;0.05). Apoptosis index of lung tissue in lymphatic duct ligation groups was significant lower than that in no-ligation groups (Plt;0.05). Levels of endotoxin, D-lactate, and diamine oxidase in lymphatic duct ligation groups had downward trends compared with no-ligation groups, but there was no statistical signification (Pgt;0.05). Conclusions Intestinal ischemia/reperfusion injury of rats can cause intestinal permeability increase, bacterial endotoxin translocation and systemic inflammatory response. Mesenteric lymphatic duct ligation and glutamine enteral nutrition intervention can weak lung tissue damage, increase thickness of intestinal mucosa, maintain intestinal barrier function, reduce endotoxin translocation and attenuate systemic inflammatory response. Enteral nutrition with glutamine was better than normal enteral nutrition.
Objective To study the effects of glutaminase (GA) gene blocked by antisense nucleotide on apoptosis of transplanted gastric carcinoma cells in nude mice. Methods The plasmid containing antisense sequence of GA gene was trans-fected into gastric carcinoma cells , then the cells were injected to endermic tissue of nude mice to create animal models of gastric carcinoma. Apoptosis of tumor cells was detected by terminal deoxynucleotidyl transferase2mediated nick end labeling (TUNEL) method. The expression of GA mRNA in tumor tissue was measured by reverse transcription polymerase chain reaction (RT2PCR) technique. Results After the successful transfection of plasmid containing antisense sequence of GA gene into gastric carcinoma cells , the tumor’s growth speed decreased , apoptosis of tumor cells increased , and the expression of GA mRNA also decreased. Conclusion The antisense gene of GA could inhibit the expression of GA gene and significantly increase the apoptosis of gastric carcinoma cells.
Objective To observe the effect of glutamine （Gln） on intestinal permeability after surgery of children, also its influence on the plama level of interleukin-2（IL-2）, endotoxin and synthesize of protein through a random nutrition trial. Methods Twenty children suffered from congenital heart disease were divided into Gln group and control group with random number table, 10 cases in each group. They were all given isonitrogenous and isocaloric total paraenteral nutrition after 24 h postoperatively. In Gln group the Dipeptiven [-N （2）-L-alanyl-Lglutamine] was used with 2 ml/kg · 24h additionly. Before operation, 24h and 96 h after operation, intestinal permeability, serum level of endotoxin, IL-2, C-reaction protein, prealbumine were measured. Results Intestinal permeability increased in 24 h after cardiac surgery in two groups, while the concentration of endotoxin also increased, 96 h after surgery the intestinal permeability recovered, but the endotoxin level did not decrease in control group （P〈0. 01）. Conclusion Utilization of Gln can improve immune suppression, elevate the IL-2 level, decrease the endotoxin concentration, alleviate the infection, but has no effect on the protein synthesis after congenital cardiac operation of children.
Objective To study glutamine (GLN) and cholecystokinin (CCK) effects on prevention of cholestasis in total parenteral nutrition (TPN). Methods White rabbits were choosed as TPN models, which were divided into four groups, group 1, TPN only (n＝10); Group 2, TPN plus GLN administration (n＝10); Group 3, TPN plus CCK (n＝10); Group 4, TPN plus GLN and CCK administration. Bile components were assayed and the structural change in gallbladder and liver were observed under light and election microspes at the forth and eighth week. Results Increasing of bilirubin and cholesterol was observed in the 1st and 2nd groups at the forth week, but increasing in the 3rd group was observed at the eighth week. The 4th group was normal. Changes of gallbladder and liver structure in 1st and 3rd group occured at the forth week. Changes of 2nd group occured at the eighth week. No structural change was found in the 4th group. Conclusion The test prove that cholestasis would occure during TPN and become serious with time prolonging. Integrity and function of gallbladder-wall tissue cell could be defended and sustained by applying GLN, but there is no direct preventing action. There is apparent cholecy stokinetic and cholagogic fundations by applying CCK. But CCK would lose its function if gallbladderwall was damaged. The test prove that TPN+GLN+CCK is the best way to prevent cholestasis during TPN.
【Abstract】 Objective To study the effects of glutamine (Gln) combined with growth hormone （GH） on the levels of cytokine (TNF-α, IL-1, IL-6), coritsol and amino acid metabolism in septic rats. Methods Ten out of 79 SD rats were randomly collected as the control group. Thirty of 69 septic SD rats, which were made by cecal ligation and perforation (CLP) method and were given parenteral nutrition (PN) lived to day 6. They were also randomly divided into three groups as follows: septic group (n=10), parenteral supplemented glutamine group (Gln group, n=10), and Gln combined with GH (Gln+GH group, n=10). On the 6th day, blood drew from portal veins of the dead rats was used to detect the levels of TNF-α， IL-1， IL-6 and cortisol by ELISA. The plasma concentrations of free amino acids were determined by amino acid auto-analyzer. The muscle tissue of extensor digitorum longus was used to determine 3-methyl-histidine (3-MH) by high performance liquid chromatographic (HPLC). Results Except for the control group, most rats developed celiac abscess, hepatic abscess and pulmonary infection. The serum levels of TNF-α, IL-1, IL-6 and cortisol were significantly higher in the septic group than those of the other three groups, and they were significantly lower in the Gln+GH group than those of the Gln group, P＜0.05. Compared with the other three groups, the concentration of total amino acid in the septic group increased more, among which the glutamine and the branched chain amino acids were prominent. Most of concentrations of the amino acids decreased in the Gln group and the Gln+GH group, and the decreased amplitude of the Gln+GH group was larger, P＜0.05, albeit its level of Gln markedly increased. The concentration of 3-MH in muscle tissue was the highest in septic group, and it was significantly higher in the Gln group than that of the Gln+GH group, P＜0.01. ConclusionIt may be necessary to supplement GH combined with Gln as the content of PN to decrease cytokine levels and im-prove amino acid metabolism for septic case.
Objective We investigated the effect of supplementation with alanyl-glutamine dipeptide on insulin resistance and outcome in patients with chronic obstructive pulmonary disease (COPD) and respiratory failure. Methods A prospective, randomized, open and controlled trial was conducted. Patients with COPD and respiratory failure were recruited between Jan 2005 to Feb 2006 and randomly assigned to a trial group (n=14) with glutamine dipeptide supplmented parenteral nutrition and a control group (n=16) with isocaloric, isonitrogenic parenteral nutrition. On the third day and fifth day of nutrition treatment, blood glucose was clamped at level of 4.4 to 6.1 mmol/L by intravenously bumped insulin. Blood gas, blood glucose level, insulin dosage were recorded everyday. The outcomes were mortality, length of stay (LOS) in hospital and in ICU, mechanical ventilation times and the costs of ICU and hospital.Results Thirty patients successfully completed the trial. There was no difference in blood gas between two groups, but PaO2 increased gradually. Compared with control group, blood glucose level had trend to decrease in trial group. The average insul in consumption decreased significantly in trial group on the fifth day. There was no statistical difference between two groups in mortality, length of stay in hospital and the costs of hospital. But compared with control group, length of stay in ICU and mechanical ventilation days had trend to decrease in trial group. Conclusion Alanyl-glutamine dipeptide do not improve pulmonary function of patients with COPD and respiratory failure. However, alanyl-glutamine dipeptide attenuated insul in resistance and stabilized blood glucose. This trial does not confirm alanyl-glutamine di peptide can improve outcome in critically ill patients with COPD and respiratory failure between two groups in mortality at the end of 30 days, length of stay in hospital and the costs of hospital. But the length of stay in ICU and the duration of mechanical ventilation does decrease, but not significantly, in the trial group.
Objective To investigate the joint effects of selective digestive decontamination (SDD) and glutamine (Gln) on preventing intestinal bacterial translocation of orthotopic piggyback liver transplantation and to observe the incidence of postoperative pneumonia in rabbit. Methods Thirty rabbits received orthotopic piggyback liver transplantation and were randomly divided into three groups (SDD group, SDD+Gln group and control group). Mixed emulsion of tobramycin, polymyxin E and nystatin were given to the rabbits in SDD group. Same dosage of the above components plus Gln were given to the rabbits in SDD+Gln group. Samples of portal vein blood, ileum tissue and lung tissue were obtained in each group at different phases during and after operation, the pathological changes of ileum tissue, the bacterial translocation in blood of portal vein and the incidence of postoperative pneumonia were detected. Results The mixing section area of intestinal blood capillaries in SDD+Gln group was smaller compared with control group (P＜0.05， P＜0.01) and SDD group (P＜0.05) while the portal vein was obstructed for 15, 30 and 45 min, and after the operation, respectively. The length of ileum villus in SDD+Gln group was longer than that in control group (P＜0.05) and in SDD group (P＜0.05) before the portal vein was obstructed, but the length of ileum villus in control group gradually became longer and eventually exceeded that in SDD+Gln group at the time of 45 min after the portal vein was obstructed (P＜0.05). After the operation, the length of ileum villus in SDD+Gln group was significantly longer than that in SDD group (P＜0.05) and control group (P＜0.01). At the time of 45 min after the obstruction of portal vein and 30 hours after operation, the positive rate of cultured bacterial in the blood of portal vein in SDD+Gln group was significantly lower than that in control group (P＜0.05, P＜0.01). The incidence of postoperative pneumonia in SDD+Gln group and SDD group were significantly lower than that in control group (P＜0.05,P＜0.01). Conclusion Gln could nourish intestinal epithelium of mucous membrane.When combined with SDD, it could decreased the incidence of intestinal bacterial translocation occurred during the obstruction of portal vein and after operation, so as to decrease the incidence of postoperative pneumonia.
Objective To construct gene silence adenovirus vector targeting both transglutaminase 2 （TG2） and Mer receptor tyrosine kinase （Mertk） synchronously and detect the gene silence function of it. Methods The interfering plasmids targeting TG2 protein and Mertk protein were constructed firstly， then the H1 promoter and RNA interfering （RNAi） sequence were cut and ligated to pAdTrack for constructing pAdTrack/TG2/Mertk. The pAdTrack/TG2/Mertk was transfected into BJ5183 bacterial cells which contained pAdEasy-1， then the plasmid was detected by enzyme digestion after recovery. Adenovirus were harvested after that pAdTrack/TG2/Mertk was infected into HEK293 cells. The virus titer was measured after repeated amplification. The RAW264.7 cells were infected by pAdTrack/TG2/Mertk， pAdTrack/TG2， pAdTrack/Mertk， and pAdTrack/green fluorescent protein （GFP）， respectively. Then the expression levels of TG2 protein and Mertk protein of mouse macrophages were detected by Western blot after infection. Results The virus titer of pAdTrack/TG2/Mertk plasmid was 6.13×1010GFU/mL. The pAdTrack/TG2/Mertk plasmid which contained 2 promoters and 2 RNAi sequences was identified successfully by enzyme digestion. Compared with pAdTrack/GFP group and pAdTrack/Mertk group （there was no significant differece between the 2 groups）， the expression levels of TG2 protein of mouse macrophages which infected with pAdTrack/TG2/Mertk or pAdTrack/TG2 decreased obviously （P＜0.01）， but there was no significant difference between the later 2 groups. Compared with pAdTrack/GFP group and pAdTrack/TG2 group （there was no significant difference between the 2 groups）， the expression levels of Mertk protein of mouse macrophages which infected with pAdTrack/TG2/Mertk or pAdTrack/Mertk decreased obviously too （P＜0.01）， but there was no significant difference between the later 2 groups. Conclusion Gene silence adenovirus vector plasmid targeting both TG2 and Mertk synchronously is constructed successfully， and the pAdTrack/TG2/Mertk can reduce the expressions of TG2 protein and Mertk protein of mouse macrophages obviously.
ObjectiveTo evaluate the effect of glutamineenhanced enteral nutritional support on elder patients after total gastrectomy. MethodsA total of eightyfour cases of elder patients receiving total gastrectomy were included in this study from February 2008 to August 2010. The patients were randomly divided into three groups: glutamineenhanced enteral nutrition (Gln) group, enteral nutrition (EN) group and parenteral nutrition (PN) group. The complications and hospital stay after operation were compared, and the levels of serum total protein, albumin, proalbumin， and transferrin of patients were measured before operation, on 2 d and 10 d after operation, respectively. Furthermore, the percentage of CD4 and CD8 T cells, CD4/CD8 ratio， and the levels of serum IgM and IgG of patients in peripheral blood before and after operation were detected. ResultsNutritional therapy was successfully performed in patients of three groups. The anal exhaust time and hospital stay after operation of patients in Gln group and EN group were significantly lower than those in PN group (Plt;0.05). The difference of postoperative complications and digestive tract symptoms of patients in three groups was not obvious (Plt;0.05). Anastomotic fistula occurred in one patient of PN group on 6 d after operation and was cured by conservative treatment for 54 d. The difference of total protein, albumin, proalbumin, and transferrin levels of patients in three groups before operation was not significant (Pgt;0.05), and these indexes fell dramatically on 2 d after operation and lower than before operation (Plt;0.05), although the intergroup difference was not statistically significant (Pgt;0.05). On 10 d after operation, all indexes recovered in different extent, while those data in Gln group and EN group were significantly higher than those on 2 d after operation (Plt;0.05). The levels of total protein, albumin, and proalbumin of patients in Gln group and EN group were markedly higher than those in PN group (Plt;0.05), although there was no difference between the former groups (Pgt;0.05). The difference of several immunological parameters of patients in three groups before operation was not significant (Pgt;0.05). On 10 d after operation, the percentage of CD4 and CD8 T cells, CD4/CD8 ratio, and the levels of serum IgM and IgG of patients in Gln group returned and even exceeded the preoperative results, which were significantly higher than those in EN group and PN group other than IgM (Plt;0.05). The postoperative results of all parameters except IgG in EN group were significantly lower than preoperative results in patients of EN group and PN group (Plt;0.05). ConclusionsIt is safe and feasible to elder patients who had received total gastrectomy and perioperative glutamine-enhanced nutritional support, which can improve nutrition and immune status, promote the recovery and reduce the duration of hospital stay, and nutritiional support after total gastrectomy is one of the optimal choices for these patients.