| Part one:Feasibility Study of Measuring Glutamine Concentration in Skeletal Muscle by Non-invasive Method of MRSObjective 1. To study the feasibility and possibility of measuring glutamine (GLN) level in Zelanian Rabbits' Skeletal Muscle by Non-invasive Method of magnetic resonance spectroscopy (MRS). 2. To study the feasibility and possibility of measuring GLN level in healthy adults' Skeletal Muscle by Non-invasive Method of MRS. Methods 1. 22 Zelanian Rabbits were admitted(female 11; male 11). The Zelanian Rabbits were 5-6 months old, average weight was 2.59±0.39kg. PRESS sequence single voxel was used to locate on the latter right limb skeletal muscle, 1cm above knee joint. MRS parameter set: TE=94ms, TR=2000ms, NEX=8, VOXEL=1×1×1cm~3, TIME=4min. The peak height and peak area of GLN+ GLU(GLx) was at 3.8ppm and creatinine(Cr)was at 3.0ppm by MRS scan. Then we got the data of ratio of peak height GLx/Cr and ratio of peak area GLx/Cr. Secondly, muscle biopsy was made at the same site, then the actual level of GLx was measured by High performance Liquid Chromatography (HPLC). 2. 36 healthy adults were admitted(male 18, female 18). The average age was 28.7±4.1y in male, 25.0±2.2y in female. The average weight was 67.6±7.4kg in male, 54.2±3.9kg in female. BMI was 23.2±2.6 in male, 20.1±1.3 in female. PRESS sequence single voxel was used to locate on the right leg skeletal muscle, 15cm above knee joint. MRS parameter set: TE=94ms, TR=2000ms, NEX=8, VOXEL=1×1×1cm~3, TIME=4min. The peak height and peak area of GLN+ GLU(GLx) was at 3.8ppm and creatinine(Cr)was at 3.0ppm by MRS scan. Then we got the data of ratio of peak height GLx/Cr and ratio of peak area GLx/Cr. 3. The data of peak height GLx/Cr ratio and peak area GLx/Cr ratio by MRS and date of GLx/plasma Cr by HPLC were analyzed in correlate and regression method.Results 1. The mean ratio of peak height of GLx/Cr of 22 Zelanian Rabbits was 0.162±0.045 by MRS. The mean ratio of peak area of GLx/Cr of 22 Zelanian Rabbits was 0.092±0.065 by MRS. The average concentration of GLx in skeletal muscle at the same site was 4.19±2.5μmol/g by means of HPLC. The mean ratio of GLx/ Cr (plasm) was 4.576±0.599. Relativity was analyzed between the ratio of peak height GLx/Cr by MRS and GLx/Cr(plasm) by HPLC, r=0.7 (p=0.001), R~2=0.5, regression formula was Y=-1.550+37.783X(X is independent variable, represents the ratio of peak height GLx/Cr by MRS; Y is dependent variable, represents the ratio of GLx/plasm Cr by HPLC). Relativity was analysed between the ratio of peak area GLx/Cr by MRS and GLx/Cr(plasm) by HPLC, r=0.6 (p=0.001), R~2=0.42, regression formula was Y=2.189+25.927X(X is independent variable, represents the ratio of peak area GLx/Cr by MRS; Y is dependent variable, represents the ratio of GLx/plasm Cr by HPLC). 2. The mean ratio of peak height of GLx/Cr in 36 healthy adults' skeletal muscle by MRS was 0.259±0.074, 0.230±0.06 in male, 0.289±0.07 in female. The mean ratio of peak area of GLx/Cr in 36 healthy adults' skeletal muscle by MRS was 0.243±0.12, 0.209±0.11 in male, 0.277±0.13 in female. Conclusion It is feasible to some extent to measure GLN level in skeletal muscle by non-invasive method of MRS. But there are some scarcities of MRS to measure skeletal muscle GLN level such as not very high specialist and not perfect stability. It is necessary to improve specialist and stability of MRS. Part two:Effects on Glutathione of Patients Received Glutamine Dipeptide Enriched Parenteral Nutrition Post Abdominal Surgery[Background: Glutathione (GSH) is a tripeptide compound with an important antioxidant role in the human body. GSH level and GSH/GSSG ratio reflect upon the body's present antioxidant status. Glutamine(GLN) is an essential GSH synthesis precursor conditional amino acid. Objective: 1. Peri-operative observation of RBC and plasma GSH level changes, GSH/GSSG changes, immune and liver functional changes of abdominal surgery patients; 2. To observe the influence of GLN enriched parenteral nutrition (PN) peri-operatively upon GSH level, GSG/GSSG level, immune function(IgM, IgG, IgA; CD4. CD8, CD4/CD8), liver function(ALT, ALB), infection induced morbidity and total hospitalization period of abdominal surgery patients. Methods: A prospective, randomized blind study consisting of 40 patients with abdominal surgical disease undergoing elective surgical treatment were randomly divided into two groups (the study group, the control group). All patients received total parenteral nutrition(TPN) for up to 7 days peri-operatively. Both groups received PN from 1 day preoperative to 6 days postoperative. The two groups received isonitrogen and isocaloric nutrition. Nonprotein caloric intake is 20kcal/kg/day; glycogen: lipid ratio is 7∶5; nitrogen is 0.16g/kg/day; calories and nitrogen intake ratio is 117kcal: 1g. The control-group received TPN absent of GLN dipeptide while the study-group received TPN including GLN dipeptide (20% Depipetiven 100ml, containing ALA-GLN 20g, GLN 13.46g, ALA8.2g). Plasma and RBC GSH levels, GSH/GSSG ratio, biochemical parameters(ALT, ALB, lymphocyte counts), immunoglobins (IgG, IgM, IgA) and T lymphocyte subgroups (CD4, CD8, CD4/CD8 ) were measured 1 day preoperatively and the 1st, 3rd, 6th days postoperative. Results: Control Group plasma and RBC GSH levels decreased postoperatively with obvious decrease in plasma GSH 1st, 3rd and 6th day postoperatively (4.94±1.18 umol/L; 4.90±1.20 umol/L; 5.15±0.95 umol/L) (P＞0.05): RBC GSH levels were also obviously decreased on the 1St, 3rd and 6th day postoperatively (6.47±2.14uM/g; 6.37±2.79 uM/g; 6.70±3.27 uM/g) (P＞0.05): No significant decrease of plasma and RBC GSH levels were observed in the study-group. However an increasing trend was noted in study- group RBC GSH levels days 3 and 6 postoperatively (8.19±3.44 uM/g; 7.89±1.70 uM/g), increasing by 14% and 9% respectively. Study-group plasma GSH/GSSG ratio (52.53±11.46) is higher than that of control-group (31.43±7.27 ) on the 3rd day postoperatively, (P=0.001). The alanine aminotransferase(ALT) index of both groups increased slightly postoperatively, no significant differences were observed between the two groups. Albumin(ALB) index of both groups decreased slightly postoperatively, study-group ALB levels (3.77±0.38g/dl) were higher than the control group (3.38±0.42g/dl) on the 3rd days postoperatively, (P=0.02). Lymphocyte counts of both groups decreased postoperatively, no significant differences were observed. No significant differences were noted between immunoglobin (IgG,IgM,IgA ) and T lymphocyte subgroup(CD4, CDS, CD4/CD8) levels of both groups postoperatively. There was none wound infection in the study group, while one in the control group. There was shorten trend of hospital stay in the study group compare with the control group, (22.3±2.1d vs 24.9±1.7d P＞0.05). Conclusion: 1. Stress induced by abdominalsurgical operative procedures results in decreasing RBC and plasmaa GSH levels, GSH/GSSG level, ALB, immunoglobin and lymphocytes while increasing ALT levels postoperatively. 2. Glutamine enriched TPN attenuates to decreasing degrees of plasma GSH level in patients having undergone abdominal surgery, maintains RBC GSH levels, GSH/GSSG ratio and ALB levels, thus it is likely to benefit and enhance protection of liver function post surgery, shorten...
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