Objective:To investigate the impact of alanyl-glutamine(Ala-Gln) on outcome in radiation enteritis rats.Methods: Male SD rats(n=70)were separated randomly into four groups: control group(n=10),AR+pseudosurgery group(n=20),AR+TPN group(n=20) and AR+TPN+Ala-Gln group(n=20).Rats were observed for mortality,changs of body weight,villous hight and area,the bcteriral translocation(BT)in mesenteric lymph nodes(MLNs),liver,spleen and peritoneal cavity.Serum TNF-? and sIL-2R level were determined by sandwich ELISA.Results: When Ala-Gln was administered in radiation enteritis rat,the mortality,body weight loss and bacterial translocation were decreaded,the villous hight and area was increased and the TNF-? and sIL-2R levels were reduced.Conclusion: Parenteral Ala-Gln nutrition can improves the results of radiation enteritis rats.

BACKGROUND: A latest research indicates that γ~δ T cells following touching with microbe products show characteristics as dendritic cells and antigen-presenting cells (APC) and induce intensive immune response of CD4~+ CD8~+ γ~δ T cells.OBJECTIVE: To verify APC-like functions of γ~δ T cells during transplantation rejection, investigate a simple and effective method to amplify γ~δ T cells in vitro, and to infect γ~δ T cells with FasL retrovirus system.DESIGN, TIME AND SETTING: An animal experiment was performed at Central Laboratory of the First Affiliated Hospital of the Fourth Military Medical University of Chinese PLA from August 2007 to August 2008. MATERIALS: A total of 60 healthy adult Wistar rats of clean grade and weighing 200-320 g were used to establish donor and receptor models with segmental heterotopic small intestine transplantation.METHODS: Models of segmental heterotopic small intestine transplantation were established using three sleevelet vascular anastomosis. γ~δ T cells were obtained using flow cytometry and its function was demonstrated. Mononuclear cells were routinely separated and amplified with Mtb-Ag. MAIN OUTCOME MEASURES: Cell proliferation was observed; percentage of γ~δ T cells for lymphocytes was detected using TCRγ~δ magnetic beads kit; γ~δ T cells following transfection were detected using pLXSN FasL retrovirus system. RESULTS: Activated γ~δ T cells showed dendritic cell-like adhesion function and APC-like functions during transplantation rejection. γ~δ T cells accounted 4.5% for mononuclear cells, the purification of γ~δ T cells was up to 72.2% on the 10~(th) day after activated by Mtb-Ag, and the purification of γ~δ T cells was up to 99.1% through positive magnetic sorting. 285 bp FasL fragment demonstrated that the gene integration was observed in PA317 cells. The ration of FasL+ cells was 97.3% after infected by pLXSN FasL retrovirus system.CONCLUSION: This study demonstrated APC-like functions of γ~δ T cells during transplantation rejection. γ~δ T cells were successfully amplified in vitro. PA317/ pLXSN2FasL+ retrovirus system was successfully constructed and γ~δ T cells were modified by FasL retrovirus system.

Objective:To investigate appropriate accommodation of nutrition after living-related small bowel transplantation.Methods: According to the function of allograft and general body state,TPN was used and gradually transferred to PN+EN,finally to TEN.The clinical and laboratory nutrition markers in 4 recipients were observed.Results: 2 recipients survived over 4 years,various kinds of nutrition markers were normal,and the health status was good.One recipient died of acute pulmonary infarction at 19 days.Another recipient died of multiple system organ infection at 5 months.Conclusion: EN cand promote restoration of allograft function.

OBJECTIVE To study the antibiotic resistance gene of qacE△1-sul1 in Acinetobacter baumannii isolated from inpatients in Urumqi and the distribution and difference of qacE△1-sul1 genotype in Xinjiang. METHODS Bacterial strains were identified by system of API and antibiotic susceptibility test was detected by K-B and microdilution methods according CLSI.PCR was used to determine the gene of qacE△1-sul1 and compared with the sequences in GenBank. RESULTS Antibiotic resistance phenotypes showed the resistance rate to cefataxime and tetracycline was the highest(100.0%),but that to cefoperazone/sulbactam was the lowest.The positive rate of qacE△1-sul1 gene in A.baumannii was 52.2%. CONCLUSIONS High resistance in A.baumannii is found in Xinjiang.The positive rate of qacE△1-sul1 is high.Strengthening epidemiology surveillance of resistance carried by integron-carrying gene is important.

Objective To investigate the expression of ligands of DNAM-1 and NKG2D in the colonic cancer.Methods The colonic cancer tissue and adjacent normal colonic tissues were collected from 42 colonic cancer patients who were admitted to the Tangdu Hospital of Fourth Military Medical University from June 2010 to January 2011 were retrospectively analyzed.The expressions of CD155,CD112 and MICA/B in the colonic cancer tissues and the normal colonic tissues were detected by immunohistochemistry.The expressions of CD155,CD112 and MICA/B in the colonic cell line SWll6,SW480,SW620 and Colo205 in the Duke's A,B,C and D phases were detected by cell cytometry.The relationship of the expressions of the 3 ligands and the clinicopathological parameters was analyzed using the Mann-Whitney U test,chi-square test and Fisher exact probobility.Results Week expression of CD155 was found in the normal colonic tissues,while the expressions of CD112 and MICA/B were not found.In the colonic cancer tissues,the expressions of CD155,CD112 and MICA/B were 81.0％,52.4％ and 47.6％,which were significantly increased.The expressions of CD155,CD112 and MICA/B were not correlated with the gender,tumor differentiation,lymph node metastasis and Duke's staging (P ＞ 0.05).The overall expression rates of CD155,CD112 and MICA/B in the colonic cancer cell line SWll6,SW480,SW620 and Colo205 were 88.9％,67.4％ and 42.3％,respectively.The overall expression of CD155 was significantly higher than CD112 and MICA/B (F =23.17,P ＜ 0.05).Conclusion CD155,CD112 and MICA/B express in the colonic cancer tissues and colonic cancer cell line SW116,SW480,SW620 and Colo205,and the expression of CD155 is the highest.

BACKGROUND: During pancreas transplantation, ischemia/reperfusion (I/R) injury can lead to many complications, which directly threaten the survival of the donor pancreas and the receptor itself, and the serious ones may result in the failure of transplantation. Ischemic preconditioning can protect the target organs in the following ischemia, which has become one of the hot spots in investigating organ transplantation.OBJECTIVE: To observe the early protective effect of ischemic preconditioning on the I/R injury of the grafted pancreas in the rat, and analyze its correlation with apoptosis.DESIGN: A randomized control animal experiment.SETTINGS: Department of Gastrointestinal Surgery, Department of Anesthesiology, Xijing Hospital of the Fourth Military Medical University of Chinese PLA.MATERIALS: Seventy male SD rats of 3-6 months, weighing 250-320 g, were used.METHODS: The experiments were conducted in the laboratory of Department of Gastrointestinal Surgery between September 2001 and April 2004.Six normal rats were taken as the control group, and 24 successful diabetic models were divided into I/R group and 1, 2 and 3-time ischemic preconditioning groups (n=18) according to the method of random number table,with 6 rats in each. The rats in the latter three groups were treated with 5-minute ischemia and 5-minute reperfusion for once, twice and three times respectively, all the rats underwent the pancreas transplantation. Twentyfour SD rats served as donors.MAIN OUTCOME MEASURES: ① Blood glucose before and after reperfusion in each group; Serum contents of tumor necrosis factor alpha (TNF-α) and nitric oxide; Activities of superoxide dismutase (SOD) and myeloperoxidase (MPD) and content of malondialdehyde (MDA) in the grafted pancreatic tissue; ② Apoptosis in the grafted pancreatic tissue observed by means of in situ end-labeling; Expressions of Bax and Bcl-2 proteins in the grafted pancreatic tissue with the method of Western blotting.RESULTS: ① Changes of blood glucose before and after reperfusion: The levels of blood glucose were decreased as compared with those before reperfusion in the I/R group and ischemic preconditioning groups. It was significantly lower in the 2-time ischemic preconditioning group than in the I/R group, 1 and 3-time ischemic preconditioning groups (P ＜ 0.05). ②Serum content of TNF-α at 2 hours after reperfusion: It was lower in the ischemic preconditioning groups than in the I/R group; It was lower in the 2-time ischemic preconditioning group than in the 1 and 3-time ischemic preconditioning groups (P ＜ 0.05). ③ Serum content of nitric oxide after reperfusion: It was higher in the ischemic preconditioning groups than in the I/R group; It was higher in the 2-time ischemic preconditioning group than in the 1 and 3-time ischemic preconditioning groups (P ＜ 0.05). ④SOD activity in the grafted pancreatic tissue after perfusion: It was higher in the ischemic preconditioning groups than in the I/R group; It was higher in the 2-time ischemic preconditioning group than in the 1 and 3-time ischemic preconditioning groups (P ＜ 0.05). ⑤ MAD content and MPD activity in the grafted pancreatic tissue after perfusion: Those were lower in the ischemic preconditioning groups than in the I/R group, also lower in the 2-time ischemic preconditioning group than in the 1 and 3-time ischemic preconditioning groups. ⑥ Apoptosis in the grafted pancreatic tissue: The apoptosis index after perfusion was lower in the ischemic preconditioning groups than in the I/R group; It was significantly lower in the 2-time ischemic preconditioning group than in the 1 and 3-time ischemic preconditioning groups (P ＜ 0.05). ⑦ Expressions of Bax and Bcl-2proteins in the grafted pancreatic tissue: There was high expression of Bax protein and low expression of Bcl-2 protein in the grafted pancreatic tissue after perfusion in the I/R group; Low expression of Bax protein and high expression of Bcl-2 protein in the grafted pancreatic tissue after perfusion were observed in the ischemic preconditioning groups; In the 2-time ischemic preconditioning group, the expression of Bcl-2 protein was the highest but that of Bax protein was the lowest.CONCLUSION: Ischemic preconditioning can protect the grafted pancreas from I/R injury at early pancreas transplantation, which maybe correlated with the elevation of SOD activity, increase of the synthesis of endogenous nitric oxide, down-regulation of TNF-α and the alleviations of the adhesion and aggregation of polymorphonuclear leukocytes. Ischemic preconditioning can reduce the apoptosis of the grafted pancreas, and the the possible mechanism may be correlated with the alleviations of the adhesion and aggregation of polymorphonuclear leukocytes, reduce of oxygen-derived free radicals, up-regulation of Bcl-2 protein and the down-regulation of Bax protein. 5-mintue ischemia and 5-minute reperfusion for twice is the best way to induce ischemic preconditioning in rat pancreas transplantation.

BACKGROUND: Ginkgo biloba extract (GBE) has the pharmacological actions of antioxidation, eliminating free radicals and anti-platelet activating factors, it also can relieve the ischemia/reperfusion injury of various organs.OBJECTIVE: Toobserve whether GBE can relieve the ischemia/reperfusion injury of transplanted pancreas in diabetic rats or not.DESIGN: A complete randomized grouping design, controlled study.SETTINGS: Department of Gastrointestinal Surgery and Department of Anesthesiology, Xijing Hospital, Fourth Military Medical University of Chinese PLA Hospital.MATERIALS: Totally 128 male SD rats of clean grade, aged 3-6 months,weighing 250-320 g, were used. GBE was produced by Dr. Willmar Schwabe Pharmaceuti - cals (Ginaton parenteral solution, 5 mL/piece, containing 17.5 mg GBE, including 4.2 mg ginkgo flavone glycosides, batch number: 1511102).METHODS: The experiments were carried out in the laboratory of Department of Gastrointestinal Surgery from September 2001 to April 2004.① Eighty rats were injected with STZ (65 mg/kg) via penile vein, and 60 of them with fasting blood glucose exceeding 17.4 mmol/L for 2weeks were taken as the diabetic rats, and the other 48 normal rats were taken as donors. ② The 60 diabetic rats were randomly divided into two groups: ischemia/reperfusion group (n=30) and GBE group (n=30), and pancreas transplantation was performed in both groups. In the ischemia/reperfusion group, the rats were douched with 4 ℃ iced balanced salt solution containing heparin (1.5×105 U/L) for 20 minutes. In the GBE group, the recipients were given intravenous injection of GBE (1.5 mL/kg) at 1 day and 30 minutes before transplantation, and those in the ischemia/reperfusion group were intravenously injected with saline of the same volume. The donor pancreases were all reserved in 4 ℃ iced balanced salt solution containing heparin (1.5×105 U/L), the cold and hot ischemia times were kept for 180 and 15 minutes in each group to induce ischemia/reperfusion injury of transplanted pancreas. ③ Six randomly selected rats were killed at 2 days before transplantation and at 3 and 7days after transplantation respectively to detect fasting blood glucose; The activity of amylase was determined with corresponding kit provided by Nanjing Jiancheng Bioengineering Institute; Pancreas tissues were removed for hematoxylin and eosin (HE) staining; Six rats were used to observe the metabolic indexes; The other 6 rats were used to observe the survival rate within 1 month. ④ The differences of the measurement data were compared with the paired t test.MAIN OUTCOME MEASURES: ① Changes of fasting blood glucose level, metabolic indexes and activity of amylase before and after pancreas transplantation in the rat recipients of both groups; ② Pathological changes at 3 and 7 days after transplantation in the rat recipients of both groups.RESULTS: All the 60 rat as recipients finished the detections of blood glucose, food intake, water intake, urinary output and blood amylase. ①The survival rate within 1 month after transplantation was obviously higher in the GBE group than in the ischemia/reperfusion group (83%, 33%, P＜ 0.01). ② The blood glucose, water intake, food intake and the urinary output at 3 and 7 days after transplantation were obviously decreased as compared with those at 2 days before transplantation in both theischemia/reperfusion group and GBE group (P ＜ 0.05-0.01), and those at 3 and 7days after transplantation were obviously lower in the GBE group than in the ischemia/reperfusion group (P ＜ 0.05-0.01). ③ The activity of blood amylase at 3 days after transplantation was obviously increased as compared with that before transplantation in both the ischemia/reperfusion group and the GBE group (P ＜ 0.01, 0.05), it was still obviously higher at 7 days after transplantation than at 2 days before transplantation in the ischemia/reperfusion group (P ＜ 0.01), and it had almost recovered to normal in the GBE group. The activities of blood amylase at 3 and 7 days after transplantation were obviously lower in the GBE group than in the ischemia/reperfusion group (P ＜ 0.01). ④ The results of the pathological observation showed that the damaged severity of the transplanted pancreas was greater in the ischemia/reperfusion group than in the GEB group.CONCLUSION: GBE pretreatment can improve the survival rate of pancreas transplantation in rats, reduce the activity of blood amylase, ameliorate the metabolism, relieve the severity of reperfusion injury of pancreas,and plays a protective role in the pancreas transplantation.

Objective To investigate Nuclear factor-κB (NF-κB) expression in tumor associated inflammation in patients with adamantinomatous craniopharyngima..Methods Fifty-four patients (31 male and 23 female) with craniopharyngioma from 3 to 66 years of age were recruited from May 2004 to March 2006.NF-κB and Osteopontin (OPN) expression in human craniopharyngiomas were detected using immunohistochemical staining.High-sensitivity C-reactive protein (hs-CRP), a systemic marker of inflammation, was examined in patients' tumor hydatid fluid, cerebrospinal fluid and serum.Results NF-κB expression was significantly increased in the adamantinomatous craniopharyngioma.Spearman;s correlation analysis demonstrated that NF-κB expression was associated with OPN expression.The hs-CRP level was also increased in the tumor hydatid fluid (4.28±0.90 mg/mL), cerebrospinal fluid (0.035±0.006 mg/mL) and serum (1.72±0.54 mg/mL) in patients with adamantinomatous craniopharyngioma.Conclusions NF-kappa B is closely associated with tumor associated inflammation which further mediates adhesion of tumor to surrounding important structures in patients with adamantinomatous craniopharyngioma.

Objective To explore the influence of delineator and contouring criteria training on the delineation of the tumor bed and whole breast target after breast-conserving surgery.Methods Twelve brcast cancer patients after breast conserving surgery were selected.Tumor bed marked by clips was defined as gross target volume 1 (GTV1),tumor bed formed by seroma was defined as GTV2 and the whole breast was defined as clinical target volume (CTV).Five junior radiation oncologists first delineated GTV1,GTV2 and CTV for each patient following their own criteria.After contouring criteria training,they then delineated GTV1,GTV2 and CTV for the same group of patients again.The differences of the volumes of GTV1,GTV2 and CTV before and after training among different delineators were compared.One-way ANOVA or matching t-test was performed.Results The inter-delineator variability on GTV1,GTV2 and CTV delineation before training was statistically significant (F =11.16,7.54 and 3.78,P =0.000,0.000 and 0.009).After training,the inter-delineator variability on GTV1 and GTV2 delineation had statistical significance (t =4.78 and 4.24,P =0.002 and 0.005),but the inter-delineator variability on CTV delineation had no statistical significance (t =1.52,P =0.209).The coefficient of variance of the GTV1,GTV2 and CTV before and after training was significantly different (t =3.14,2.81,2.70,P =0.009,0.017 and 0.021).The matching index of GTV1,GTV2 and CTV before and after training was significantly different (F =16.08,8.61,8.48,P =0.000,0.000 and 0.000).Conclusions In delineating the target of breast cancer,application of the criteria of target delineation can reduce the difference among the delineators,especially for CTV.

BACKGROUND: Hepatic oval cells (HOCs) possess the potential of self-renewal, replication, and clone, proliferation and differentiation into mature hepatocytes under a certain condition. HOCs can be used as biomaterial for constructing biological artificial liver in vitro, employed for in vivo transplantation, as well as for tissue engineering as seed cells. HOCs can be widely used for improving clinical treatment of liver diseases. OBJECTIVE: To establish adult Wistar rat models of HOC proliferation, to perform/n vitro isolation and culture of HOCs, and to study the possibility of induction and differentiation of HOCs into hepatucytes. DESIGN: Observational study. SETTING: Institute of Gastroenterology, Nanfang Hospital, Southern Medical University. MATERIALS: Experiments were performed at the Laboratory of Institute of Gastroenterology of Nanfang Hospital from December 2003 to February 2006. Thirty-six healthy male Wistar rats aged 3-4 months (150-200 g) were provided by Experimental Animal Center of Southern Medical University. METHODS: Male Wistar rats were orally fed with ethionine received two-thirds partial hepatectomy (2/3 PH). HOCs were harvested and purified by two-steps perfusion and Percoll density gradient centrifugation, and then cultured in vitro and induced with hepatocyte growth factor (HGF), oncostatin M (OSM) and fibroblast growth factor-4 (FGF4). MAIN OUTCOME MEASURES: Identification and differentiation of HOCs. RESULTS: The concentration of HOCs was about 1.34×108 L-1 in each rat model after in vitro isolation. These cells were round, oval or polygon, about 1/6 1/3 the size of normal hepatocytes. The nucleus-cytoplasm ratio was relatively large. After 2 weeks, clone-like proliferation of HOCs could be observed. Laser scanning confocal microscopy indicated positive expression of stem cells markers Thy-1 and C-kit in cytoplasm and membrane of HOCs. Immunocytochemistry demonstrated positive stem cells marker alpha fetoprotein (AFP) in cytoplasm of HOCs. HOCs can stably passage and its shape gradually changed after inducing with HGF, OSM and FGF4. HOC volume became larger and HOCs lost their ability of sticking to the wall of culture flask. Apparent positive stain of cytoplasm albumin (Alb) was detected 14 days after induction, and the positive ratio increased along with the extension of inducing duration. Results of cytochemistry indicated a brown or black deposit after glucose-6-phosphotase (G-6-P) staining and red particles after periodic acid-Schiff (PAS) staining. CONCLUSION: Adult Wistar rat models of HOC proliferation are replicated by ethionine feeding combined with 2/3 PH. HOCs can be obtained through collagenase perfusion and Percoll density gradient centrifugation. Rat HOCs can be passaged and cultured in vitro. Under a certain condition, HOCs can be induced and differentiated into hepatocytes.