Objective To study the value of postmortem imaging on the diagnosis of intestinal perforation.Method Postmortem imaging(PMCT and PMCTA)data of 2 intestinal perforation deaths(and 4 controlled cases)were reviewed retrospectively.Diagnosing capacities of intestinal perforation by postmortem imaging method were further investigated.Results PMCT is sensitive in detecting the free air and liquid induced by intestinal perforation.PMCT can sometimes detect the gravity-dependent purulent secretions in the abdominopelvic cavity.PMCTA can visualize the extravasation of contrast agent from the perforation,which can be used to locate the accurate perforation region.Conclusion Postmortem imaging method(PMCT and PMCTA)is an important tool for the diagnosis of intestinal perforation,which can not only be used as a forensic diagnosis method,but is also useful to locate the perforation site before an forensic autopsy.

Objective To preliminarily explore the feasibility of tree shrew as a new kind of animal model in research of amblyopia,to discuss the primary visual cortex plasticity mechanism of form deprivation in tree shrew,and to provide a theoretical basis for further understanding the mechanism of amblyopia formation and recovery.Methods Sixty 30-days old tree shrews were divided into five groups,12 in each group:the group A had the right eye sutured for 1 month;the group B had the right eye sutured for 2 months;the group C had the left eye sutured for 1 month and then opened and the righ eye was sutured for 1 month,in other words,the group C was performed by alternating suture;the tree shrews of control group 1(D1) were in the same age as the the group A,but fed in normal breedingenvironment;the tree shrews of control group 2(D2) were at the same age of groups B and C,but fed with a normal diet.Samples of the visual cortex were taken after the completion of modeling,and were processed to observe the histology and ultrastructure of the visual cortex,the neuron apoptosis,and the c-fos protein expression in the tree shrews of different groups.Results Damages to different degrees were found by histological and electron microscopic examination of the visual cortex in each experimental group,and they were more obvious in the group sutured for 2 months.TUNEL staining showed that there were no significant differences between the apoptosis in the experimental and control groups.The expression of c-fos mRNA and protein in the experimental groups was decreased,and it was the lowest in the group sutured for 2 months.There was a small increase in the c-fos expression after the alternate suture,and no significant difference of c-fos expression was found in the control groups.Conclusions Different degrees of deprivation amblyopia lead to different histopathological changes.There is a plasticity in the neurons affected by amblyopia.Tree shrew can be used as an ideal animal model for the studies of form deprivation amblyopia.

Objective To evaluate the relationship between preoperative MRI characteristics and the perioperative outcomes of microvascular decompression in primary trigeminal neuralgia. Methods To analyze the relationship between preoperative MRI characteristics and the perioperative outcomes in 103 primary trigeminal neuralgia patients with micro-vascular decompression in Nanfang Hospital. The MRI features such as the ratio of CPA area, TGN cross-sectional area and TGN length was evaluated together with the TGN oppression distance, the position of TGN, the position of basilar ar-tery and the type of offending vessel as well as the outocmes including complete disappearance, temporary remission and ineffectiveness. Results Univariate analysis showed that the oppression orientation (P=0.017), oppression distance (P<0.001), offending vascular type (P=0.016), TGN cross-sectional area ratio (P<0.001) were the influencing factors of periop-erative outcomes. Logistic regression analysis showed that the offending vascular type (P=0.002)and TGN cross-sectional area ratio (P=0.020) were the main predictive factors of perioperative outcomes of microvascular decompression. Conclu-sions Preoperative thin slice MRI scanning showed that the offending artery, non-atrophy nerve roots, far distance from op-pression point to brainstem may be the favorable factors of perioperative outcomes of microvascular decompression.

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.

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.

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.

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.

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.

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.