Serum citrulline is a simple quantitative marker for small intestinal enterocytes mass and absorption function in short bowel patients.

BACKGROUND: To investigate the clinical significance of serum citrulline in evaluating the remnant small bowel enterocytes mass and absorptive function in short bowel (SB) patients. MATERIALS AND METHODS: Serum citrulline concentrations were determined using high-performance liquid chromatography (HPLC) in 22 SBS patients and 33 healthy controls. Five-hour urine D-xylose excretion and digestive protein absorption were measured using HPLC and micro-Kjeldahl method, respectively. Small bowel length and surface area were assessed on X-ray radiograph. Correlations between serum citrulline levels and small bowel length, small bowel surface, and nutritional substrate digestive absorption percentage were analyzed. For six patients receiving bowel rehabilitation therapy, serum citrulline, D-xylose excretion, and intestinal protein absorption were measured pre- and immediately postmanagement, and their correlations were analyzed. RESULTS: Serum citrulline levels were significantly lower in SB patients compared with healthy controls. In SB patients, they correlated well with remnant small bowel length (r = 0.82, P < 0.001), surface area (r = 0.86, P < 0.001), 5-h urine D-xylose excretion (r = 0.56, P = 0.007), and digestive protein absorption (r = 0.48, P = 0.046). The increased percentage of serum citrulline level in six patients receiving rehabilitation therapy followed a trend of correlating with that of intestinal protein absorption (r = 0.79, P = 0.063) and urine D-xylose excretion (r = 0.81, P = 0.053). CONCLUSIONS: In patients with short bowel syndrome, serum citrulline is a simple and accurate biomarker for the severity of intestinal failure and may be a candidate marker for the gut-trophic effects of bowel rehabilitation therapies.

Establishment of the method for determination of serum citrulline and its application in intestinal transplantation / 中华器官移植杂志

Objective To investigate the level of serum citrulline and its significance in the patients subject to intestinal transplantation. Methods Phenylisothiocyanate served as derivative reagent and the serum citrulline was determined by HPLC. Thirty-three healthy adults served as controls. Serum citrulline was measured in 2 cases of intestinal transplantation during different transplant periods. Results The serum citrulline level in 33 healthy adults was (16.87?5.97)?mol/L. Before operation, it was low (less than 10 ?mol/L) in the patients and after operation it reached to high levels and kept at higher levels. The maximium of serum citrulline was more than 30 ?mol/L in patient A and was over 50 ?mol/L in patient B. Following rejection, the serum citrulline level in patient A was decreased rapidly and simialr to that before operation and in patient B it was lower than that before operation. Conclusion The level of serum citrulline in intestinal transplantation patients was correlated with transplant rejection. More patients are reqiured for the further study.

Determination of somatostatin in serum by large pore liquid chromatography column / 肠外与肠内营养

Objective: To develop a method in the determination of somatostatin level in blood for clinical medication. Methods: A method for the determination of somatostatin(Octreotide) in serum by high performance liquid chromatography with diode-array detector(DAD) was developed.After ultrafiltration,the pH values of blood samples were adjusted to 4,and samples were analyzed by Kromasil C18 column(4.6 mm? 250 mm,5 ?m i.d.) with a large pore(300) by acetonitrile/water/trifluoroacetic acid(20/80/0.01%,v/v) as mobile phase. Results:The recovery rate of Octreotide was 85%,and the calibration curve showed good linearity in the range of 0.006-0.5 mg/mL. Conclusion: The method is suitable for the detection of Octreotide content by liquid chromatography.

Experience of thin layer chromatography separation and bioassay of platelet activating factor / 医学研究生学报

Objectives：To introduce the author's experiences in bioassay of platelet activating factor with thin layer chromatographic separation and platelet aggregation method,and to discuss probable problems in the process.Methods and Results：The method was used in determining portal venous blood samples from swine models of acute severe pancreatitis,control value were 0.79～1.65 pmol/L,in acute severe pancreatitis,mean blood level of platelet activating factor may be as high as 14.75 pmol/L.Conclusions:Thin layer chromatography separation and platelet aggregation assay were suitable for the determination of platelet activating factor,the results were satisfactory in case of standardized operation.

Peripheral amino acid level in patients with short bowel syndrome / 中国普通外科杂志

Objective To study the alterations of serum amino acid level in patients with short bowel syndrome (SBS). Methods Blood levels of 17 amino acids were measured in 17 SBS patients on admission to the hospital, and 15 healthy volunteers. Results Compared with the control group, serum concentrations of Val, Leu, Ile, Lys, Met, Ser, His, Cys in patients with SBS significantly decreased ( P

Determination of 3-methylhistidine and tyrosine in incubated buffer and checking samples by HPLC with PITC pre-column derivation / 肠外与肠内营养

Objectives:To determine 3-methylhistidine(3-MH) and tyrosine(Tyr) in standard solution and preparations by high performance liquid chromatography(HPLC) with phenylisothicyanate(PITC) pre-column derivation. Methods:PITC was used as pre-column derivative of HPLC and stored in refrigerator after drying.Two phases gradient elution was performed with acetic sodium and cyanide for 50 minites. Results:The linearity of the measurement was very good for 3-MH and Tyr(r= 0.999 1 and 0.999 2).The recovery rate was 82.6% and 87.6%,respectively. Conclusions:The method of HPLC with PITC pre-column derivation for determination of 3-MH and Tyr is stable,efficient,and sensitive.PITC is an ideal derivative.

S-Adenosyl-L-Methionine prevents total parenteral nutrition-induced cholestasis in infected rats / 肠外与肠内营养

Objectives:This study was designed to observe the effect of S adenosyl L methionine (SAMet) on total parenteral nutrition (TPN) induced cholestasis in infected rats and to analyse the change of the bile acids in the bile. Methods:18 Sprague Dawley rats were divided randomly into 3 groups,control group,infection plus TPN group,and infection plus TPN and SAMet group.Each contained 6 rats.Caecum was ligated to induce intra abdominal infection.Nutritional support was maintained for 5 days.Bile was collected by cannulating catheter.The bile flow was determined.The blood samples were gathered to measure the levels of serum total bile acid(STBA),gammaglutamyltranspeptidase (GGT),alanine aminotransferase (ALT)and alkaline phosphatase (AKP).The pathological change in the liver was observed under light microscope and electron microscope.The billiary bile acid contents were determined by high performance liquid chromatographic (HPLC) method. Results:The infection plus TPN group showed that the bile flow was reduced,levels of STBA,GGT and AKP were elevated,and fat infiltration,bile capillary dilatation and bile embolism were obvious in the liver.However,the infection plus TPN and SAMet group showed that the bile flow was increased,levels of STBA,GGT,ALT and AKP were decreased,and the liver was kept normal. Conclusions:The use of SAMet is helpful to could prevent TPN induced cholestasis in infected rats.

The measurement of vitamine B in the patients with Crohn's disease by HPLC / 肠外与肠内营养

Objective: To compare the difference of Vitamin B in serum in healthy adults and patiens with Crohn's disease.Methods: 24 healthy adults and 20 patients were measured for Vitamin B1,B6,B12 in serum.A HPLC-DAD method was developed to determine vitamin B extracted directly from human serum.Purospher RP-18 column was used for the gradient elution with mobile phase: 0.04 mol phosphate buffer: methanol(60∶40).Results: The results proved to be linear in measurements of VB1,VB6 and VB12 and the correlation coefficients were 0.999 827,0.985 097 and 0.999 994,respectively.The contents of Vitamin B(VB1,VB6,VB12) in Cronhn's patients were significantly lower than in healthy adults.Conclusion: The measurements of Vitamin B in human serum by HPLC-DAD method are clinically helpful to diagnosis and treatment for Crohn's disease.

Evaluation of the absorptive area and capacity of short bowel patients using Dxylose absorption test / 肠外与肠内营养

O bjective:To investigate the effectiveness of D-xylose absorption test in evaluating the ab-sorptive capacity of the remnant small bowel in short bowel patients.M ethods:5 hr urine D-xylose ex-cretion,1hr,2hr,3hr serum D-xylose levels and AUC was compared between the short bowel patients(n=22) and the healthy control(n=50).The correlationship among the D-xylose absorptive capacity,the remnant small bowel length and area,and protein absorptive capacity were analyzed.For the 6 pa-tients receiving rehabilitative therapy,theirD-xylose and protein absorption capacities before and after the therapy were also stud ied.R esu lts:5hr urine D-xylose excretion,1hr,2hr,3hr serum D-xylose levels and AUC were all significantly lower in short bowel patients compared to control group(P

Simultaneous determination of lactulose and mannitol excretion ratio in urine by high performance liquid chromatography / 肠外与肠内营养

Objective: Lactulose and mannitol excretion ratio in urine appears to be a marker to measure the intestinal permeability. Methods: A method has been developed for the simultaneous determination of lactulose and mannitol in urine by high performance liquid chromatography.After ultrafiltered and deionized,urine samples were analyzed with a lichrosher NH 2 column(4.6 mm?250 mm,i.d.,5 ?m),a mix of acetonitril water (72∶28,volume ratio) as mobile phase and refractive index detector. Results: The recoveries were 97.86%、 99.19% respectively and the relative standard deviation were 2.12、2.04.In 18 urine samples from healthy volunteers and patients who ingested the test solution containing 10 g lactulose and 5 g mannitol,the 6 h urinary excretion ratio of lactulose/mannitol was 0.0275?0.0113. Conclusions: It is suggested that lactulose and mannitol excretion ratio was anaccurate marker of intestinal permeability.

Nutritional support and metabolic intervention after combined liver and intestinal transplantation / 肠外与肠内营养

Objective: To observe the effect of nutritional support and metabolic intervention in a patient receiving combined liver and intestinal transplantation. Methods: Glycyl glutamine(Gly Gln)and arginine supplemented total parenteral nutrition(TPN) was administered since postoperative day(POD)1. Glutamine(Gln) and arginine supplemented enteral nutrition(EN) was applied since POD 4. Growth hormone(GH) was delivered intermittently since POD 4.With adaptation and tolerance, enteral feeding was progressively increased while parenteral nutrition reciprocally decreased. Results: Transplanted organs functioned well. Conclusions: Rehabilitation of the allograft function can profit from the application of nutritional support and metabolic intervention.

Metabolic effects of administration of branched chain amino acids on postoperative patients / 肠外与肠内营养

Objective: To investigate the metabolic effects of two different formula of amino acids on postoperative patients . Methods: According to prospective ,randomized , single blind and controlled rule, 120 patients after abdominal operation were randomized to receive parenteral nutrition with either amino acid enriched BCAA or balanced amino acid (18 F).From the second day after operation, total parenteral nutrition was infused to the patients with equal calorie and equal nitrogen by central or peripheral vein for 6 days. Meanwhile, every day nitrogen balance was monitored by collecting urine in 24 hours .The indices were investigated such as amino acids patterns, and serum levels of total protein, albumin, prealbumin, thansferrin and fibronectin. Results: Nitrogen balance of patients in study group on day5 was higher than those of control group. The decreased serum levels of albumin in study group were lower than the control group. Concentrations of valine increased more significantly in study group than that of the control group. Conclusions: The formula of amino acid enriched BCAA may be more suitable and effective for the postoperative traumatic patients on protein metabolism.

Measurement of intestinal permeability in normal Chinese volunteers / 肠外与肠内营养

Objectives:To establish a confident and sensible method of measuring lactulose(L) and mannitol(M) in human urine, and to research lactulose and mannitol excretion ratio of urine in normal Chinese volunteers in 6 hours after taking lactulose and mannitol. Methods:20 healthy volunteers took 10g of lactulose and 5g of mannitol, and 6 hours of urine was collected. The L and M concentration was analyzed by HPLC RI. The excretion of L and M and the L/M ratio was calculated. Results:The excretion of L and M in the urine was 5.74%?0.58% (0.65 %~11.47%) and 0.132%?0.012%( 0.005 %~0.206%),while the L/M ratio was 0.028? 0.010 (0.013~0.051). Conclusions: HPLC is a kind of confident and sensible method to analyze L and M concentration. The L/M ratio can reflect the intestinal mucosal permeability and can be used to monitor intestinal barrier function in clinical practice.

The effect of lipopoly-saccharide on glutamine metabolism in the rat Kupffer cells / 肠外与肠内营养

Objectives:To observe the metabolism of glutamine in the rat Kupffer cells and the effect of endotoxin. Methods:The changes of glutamine concentrations were measured before and 24 h after the in vitro cultures of the rat Kupffer cells with or without the stimulations of lipopoly saccharide. Results:Consumption and utilization rate of Gln in Kupffer cells with no LPS were (225?24)?mol/L and (9 33?0 99)% repectively.When the concention of LPS was 10.00 mg/L,consumptoin and utilization of Gln were (4 937?289)?mol/L and (73.29? 4.72 )%. Conclusions:Glutamine is an important substrate of Kupffer cell metabolism in the rest and LPS activated conditions.

An experimental study of metabolic intervention of anti-TNF antibody in intra-abdominal infection complicated by multiple organ dysfunction syndrome / 肠外与肠内营养

Objectives:To observe the effect of metabolic intervention of anti TNF antibody on the hypermetabolism occurred in intra abdominal infection(IAI) complicated by multiple organ dysfunction syndrome(MODS). Methods:Twenty rabbits were operated on with cecal ligation plus puncture(CLP) inducing IAI and MODS and were randomly divided into two groups, one receiving the anti TNF serum raised against TNF ?(anti TNF group) at 0.5?h after CLP and another receiving the non specific serum (control group). All animals were placed in metabolic cages and maintained with intravenous infusion for the observation period of one week. Serum levels of cytokines(TNF, IL 6), hormones (cortisol, insulin, glucagon), biochemical indexes (glucose, cholesterol, triglyceride, albumin) and daily excretions of urea nitrogen (UN),creatinine (Cr) and 3 methylhistidine (3 MH) were dynamically determined for 7 days. The death of animals was also recorded. Results:Compared with the control group, the levels of serum TNF, IL 6 and cortisol were significantly decreased and the levels of insulin and glucagon were kept normal after the injection of immune serum in anti TNF group, with significant improvements of biochemical indexes and decreased excretions of UN, Cr and 3 MH in urine. The survival rate was significantly increased in the anti TNF group. Conclusions:The anti TNF antibody can attenuate the metabolic abnormalities of IAI and MODS, being of the metabolic intervention on the hypermetabolism.