Objective:To probe into the expression and the clinic significance of LEA on colorectal carcinoma. Methods:The expression of LEA and CEA in 140 colorectal cancer specimens and 100 colorectal non-cancerous specimens had been detectd by immunohistcchemistry S-P method.Results:The expression of LEA was relative to tumor differentiation degree and exhibits higher selectivity in high differentiation ade-nocarcinoma ( P 0.05) . The positive rate of LEA in adenoma was much higher than surrounding non-cancerous mucosa and normal mucosa. In normal mucosa the positive rate of LEA was obviously lower than that of CEA ( P

Objective　To observe the ER ultrastructures in humn embryonic epithelial cells of colonic mucosa、renal tubule and hepatocytes and also their alterations in embryogenesis.Methods　Transmission electromicroscopy technique.Results　With the embryonic development, the ER increased in amount, became complex in structure and with its characteristic ER structures in different cells. Conclusion　The changes of ER structures are one of the characters during cell differentiation.

BACKGROUND: The islet cell transplantation has provided a solid basis for diabetic therapy, but the insufficient donor limits its development.OBJECTIVE: improving the method of isolating and purifying islets to observe the transplantation effect.DESIGN: A laboratory animal research.SETTING: Key Laboratory of Animal and Department of Cell Biology, China Medical University.MATERIALS: The experiment was carried out in the Key Laboratory of Animal and Department of Cell Biology in China Medical University between January and October in 2006. Donors were Wistar rats of either gender, weight 250-300 g;Acceptors were SD male rats, weight 180-220g. The two kinds of rats were all common closed population and from the Experimental Animal Department of China Medical University (The Admission Number of Experimental Animal Institute is SYXK(LIAO)2003-0013).METHODS: ①Isolation and exaltation of islet cells as well as the functional evaluation of pancreas: After etherisation, the Wistar rat without fasting was executed. A little cut was made on the beginning of the biliary pore, then the little cut lumbar anesthesia ductus, which were connected with a 1-mm-diameter syringe and full of cold collagenase solution (1.5 g/L), was inserted directly to dilate pancreas thoroughly. The pancreatic gland was isolated and digested in the water of centrifuge, when doing that, 1 mol/L NaOH was put interruptedly into the centrifuge tube to keep the pH value of the solution at 7.8±1.0. The rat pancreas purified by centrifugation of Ficoll density gradient: The identification of purified islets was evaluated by dithizone staining. The viability of islet was assessed by fluorescence staining of aridine orange and propidium iodide. The motility rate=the total number of live cells/(the total number of live cells + the total number of dead cells)×100%. Pancreatic activity was calculated: insulin release index=the level of insulin at the third hour (high concentration glucose)/the level of insulin at the second hour (low concentration glucose). ②The blood from vena caudalis of SD rat was sampled and measured the blood sugar after the intraperitoneal injection of streptozocin. The rat was diagnosed as DM when blood sugar was more than 16.7 mmol/L twice without fasting. The DM rats were divided into two groups, every group 8 rats. The experimental group rats were injected about 1 000 islet cells into the location below renal capsule, and the control group rats were injected the same volume of 1640 cultu re solution. Eight normal rats, whose glucose concentration ≤ 5.5 mmol/L, were taken randomly as normal controlled group. The blood sugar was measured every day after the surgery. The blood sugar less than 11.1 mmol/L without fasting was taken as the sign of successful islet transplantation. Intravenous sugar tolerance test was applied to the rats of normal control group, DM control group and experimental group 3 days after islet transplantation. Fasting for 12 hours before test, the blood sugar was measured at 0, 15, 30, 60, 90 and 120 minutes.MAIN OUTCOME MEASURES: Purity quotient, survival rate and activity of islet cells.RESULTS: All 24 SD acceptor rats were involved in the result analysis without miss.①The total number of purified islets of one pancreas was (1 150±141) in well morphology. The purity of islets was more than 95%. The viability of islets was more than 98%. ②The insulin secretion response to glucose challenge in vitro showed the mean value of insulin in the low-glucose medium was (70.5±6.9) mlU/L, while that of high-glucose medium was (321.4±11.6) mlU/L, the insulin release index was 4.6±0.52, that meant the beta cell of islet functioned well. ③The blood glucose level and the insulin level in plasma of the transplanted recipients restored to normal 3 days after transplantation. The survival period of transplanted islets was (6±2) days. But there was not any change in the concentration of blood sugar in the control group (16.7 mmol/L). The intravenous glucose tolerance test showed the identical outcome between the islet splantation group and the normal control group.CONCLUSION: There are high yield and high purify of islet cells in rats, which are isolated by in situ perfusion and purified by Ficoll density gradient centrifugation.

Objective: To study effect of small ubiquitin related modifier protein 1 (SUMO1) in inflammatory reactions mediated by tumor necrosis factor (TNF)-α and nuclear factor (NF)-κB in myocardial damage of rats with type 2 diabetes mellitus (T2DM). Methods: A total of 20 Goto-Kakizaki (GK) rats with spontaneous diabetes mellitus (DM) were randomly divided into group DM1 (pure DM group, n=10) and group DM2 (DM+high-fat diet group, n=10), and another 10 normal Wistar rats were regard as healthy control group. Expressions of SUMO1, TNF-α and NF-κB were measured by immunohistochemical method. Results: 1. Levels of blood glucose and TG in group DM1 and group DM2 were significantly higher than those of healthy control group, and those of DM2 group were higher than of DM1 group ,P<0.05 all; 2. Myocardial cells lined up in order and there was no hypertrophy in group DM1; but those in group DM2 showed cells loosely lined up and hypertrophy under light microscope; 3 Immunohistochemical assay indicated that expression of SUMO1 in group DM2 and DM1 group were significantly higher than those of healthy control group [(44.5±1.1) vs. (27.2±2.2) vs. (21.7±3.0)], and of group DM2 was significantly higher than that of DM1 group (P<0.01 all); expression of TNF-α in group DM2 and group DM1 were significantly higher than that of healthy control group [(27.5±1.5) vs. (20.2±2.7) vs. (13.1±1.6)], and of DM2 group was significantly higher than that of group DM1 (P<0.01 all)；expression of NF-κB in group DM2 and group DM1 were significantly higher than that of healthy control group [（30.1±1.7）vs.40.7±1.5）vs.（16.0±2.6）], but of group DM1 was significantly higher than that of group DM2 (P<0.01 all). Conclusion: There are obvious metabolic disorders of glucose and lipid in T2DM rats, and complicated morphological changes of myocardial tissues similar to myocardial lesions in DM humans; the expressions of SUMO1, NF-κB and TNF-α significantly increase, suggest SUMO1 takes part in inflammatory reaction mediated by NF-κB, TNF-α in myocardial lesion of rat with T2DM，and may inhibit NF-κB, possesses effect of protect myocardium.

Objective: To investigate the apoptosis of renal cells induced by tumor necrosis factor alpha (TNF-α) and nuclear factor-κB (NF-κB) in diabetic rats and intervention of rapamycin. Methods: A total of 20 rats (Goto-Kakizaki rats) with type 2 diabetes mellitus (T2DM) were randomly and equally divided into DM model group (DM group) and rapamycin treatment group (DMR group, received rapamycin treatment after DM model was established); another 10 Wistar male rats were regard as normal control group. Apoptosis of renal cells, expression levels of TNF-α and NF-κB and levels of blood lipids, blood glucose were measured in all groups after four weeks and eight weeks. Results: Four and eight weeks After model was established, compared with normal control group and DMR group, there were significant increase in renal cells apoptosis [RCA, four weeks: (0.217±0.031), (0.272±0.031) vs. (0.545±0.031), eight weeks: (0.358±0.031), (0.350±0.031) vs. (0.811±0.031)] and expressions of NF-κBp65 [OD: four weeks: (0.160±0.027), (0.131±0.027) vs. (0.411±0.027), eight weeks: (0.232±0.027), (0.275±0.027) vs. ( 0.634±0.027)] and TNF-α [OD: four weeks: (0.242±0.027), (0.275±0.027) vs. (0.617±0.027), eight weeks: (0.385±0.027), (0.342±0.027) vs. (0.912±0.027)] in DM group (P<0.01 all). Correlation analysis indicated that there were positive correlations between renal NF-κBp65 and TNF-α, among RCA and TNF-α, NF-κBp65 (r=0.956, 0.953, 0.886，P<0.01 all).

Objective: The inhibition on the proliferation of colorectal carcinoma cell strain with different proliferative potential by ATRA was investigated in present study, which would benefit for the therapy of ATRA on colorectal carcinoma. Methods: The proliferation of LS174T and CW-2 colorectal carcinoma cell strain inhibited by ATRA was determined using cell observation, FACS and MTT methods. Results: The growth speed of LS174T cell strain was faster than that of CW-2. ATRA played a significant role on inhibiting the proliferation of LS174T and CW-2 cell strain and inducting the cell differetiation in vitro. Conclusions: ATRA could inhibit the growth of LS174T and CW-2 cell strain. ATRA could inhibit the proliferation of colorectal carcinoma cell and induce cell differentiation to some extent, which was correlated with the concentration of ATRA.

Objective　To prevent the after cataract induced by lens epithelial cells proliferation from postoperative cataract, monoclonal antibody (McAb) specific for rabbit lens epithelial cell is made, it will be the carrier further to be conjugated with cytotoxin. The conjugations will inhibit lens epithelial cells growth and not damage the other tissues of eye. Thereby McAb is the experimental bases of preventing after cataract.Methods　BALB/c mice were immunized by mixture of rabbit lens epithelial cells and Freund's adjuant. The immunized mouse spleen cells were fused with parental mouse myeloma cells (BALB/c　SP2/0) using polyethylene glycol (PEG-4000). The fused cells were selectively cultured by hypoxanthine, aminopterin and thymidine (HAT) culture medium. The specificities of the supernatant from hybridomas were tested by indirect immunofluorescence and immunohistochemistry SP (Streptavidin peroxdase conjugated method). The positive hybridomas were further cloned three times by methylcellulose culture medium to ensure monoclonality. At last, the consensual reaction of McAb was tested on human eye tissues.Results　Hybridomas were produced by fusion of spleen cells of immunized mice and mouse myoloma cells (SP2/0) with PEG-4000, and grown selectively in medium containing HAT after 16 days. Antibodies of the supernatant from hybridomas were tested on frozen sections of rabbit lens epithelial cell by indirect immunofluorescence. Only a positive clone secreted McAb against antigen of rabbit lens epithelial cell. The specificity of McAb was tested on paraffin sections of whole rabbit eye and whole human eye by immunohistochemistry SP. The results indicated that McAb was only positive to rabbit lens epithelial cell membrane and it was negative to the other tissues of rabbit or whole human eye tissues.Conclusions　McAb specific for rabbit lens epithelial cell was manufactured successfully. The specificity of McAb was strong. There was no consensual reaction on human eye tissues. It might be the experimental bases of further targeting chemotherapy on after cataract.

Objective To investigate the influence of menstrucal cycle and anatomic site on the fractional anisotropy (FA) values of diffusion tensor imaging (DTI) in normal breast. Methods Prospectively enrolled 96 volunteers, who have identified with normal menstrucal phase and without breast diseases were found via the breast examination, ultrasound and MRI scan. The cases were divided into three groups according to menstrucal phase: menstrual period group(menstrual cramps 1 to 6 d), proliferative phase group(menstrual cramps 7 to 14 d) and secretory phase group(menstrual cramps 15 d to the next), and each group consisted of 32 subjects. All subjects were performed bilateral breast cross-sectional T1WI, T2WI, DWI and DTI scaning. On the nipple level figture, the mammary gland was divided into three regions including the anterior, central and posterior parts, and the FA values of the different phases and regions were measured. The Kruskal-Wallis H test was applied to analyse the difference of FA values in different menstrual phase and anatomic site. Results The FA values of the anterior region in menstrual phase, proliferative phase and secretary phase were 0.21 ± 0.07, 0.24 ± 0.09 and 0.17 ± 0.07, and the difference had significant difference(P=0.014).The FA values of the central region were respectively 0.15±0.08, 0.18±0.09 and 0.15±0.07, and without the statistically significant difference(P=0.090). The FA values of the posterior region were 0.21 ± 0.11, 0.24 ± 0.13 and 0.16 ± 0.11, and also showed significant difference(P=0.002). In different regions, the difference of FA values between menstrual phases and proliferative phases were also had statistically significant(P=0.018, 0.045, respectively). In the same region, the FA value was lowest in the secretary phase, and the proliferative phase was slightly higher than menstrual phase. Conclusion The FA values are affected by menstrual cycle and anatomic site.

Objective: To study expressions of small ubiquitin-related modifier protein（SUMO）4 (SUMO4), nuclear factor (NF)- κB and inhibitory factor of NF-κB (IκB) in kidneys of rats with type 2 diabetes mellitus (T2DM). Methods: A total of ten 40-week-old male Goto-Kakizaki (GK) rats （with spontaneous diabetes mellitus）of specific-pathogen free (SPF) grade, and ten 40-week-old male Wistar rats of SPF grade were selected. The lesion of renal tissue was observed by hematoxylin eosin (HE) staining. Expressions of SUMO4, NF-κB and IκB in renal tissue were observed by immunohistochemistry methods. Results: In the GK rats, glomerular capillary ball hypertrophy, basilar membrane slightly thickening; glomerular mesangial cells hyperplasia, hypertrophy and renal tubular epithelial cells hypertrophy were observed. Compared with normal Wistar rats, expression levels of NF-κB [(0.232±0.034) vs. (0.634±0.058)], IκB [(0.242±0.027) vs. (0.712±0.078)] and SUMO4 [(0.160±0.031) vs. (0.545±0.045)] significantly increased in renal tissue of GK rats (P<0.01 all). Conclusion: Compared with Wistar rats, expressions of NF-κB, IκB and SUMO4 significantly increase in renal tissue of GK rats, suggesting that SUMO inhibiting transcriptional activity of NF-κB may exist in kidneys of T2DM rats. Therefore, sumoylation may be a new therapeutic target for inhibit renal microvascular lesion of diabetic disease.

Objective: To study relative risk factors for diabetic nephropathy (DN). Methods: A total of 238 patients diagnosed as type 2 diabetes mellitus (DM) were enrolled in the study. According to urine microalbuminuria to urine creatinine ratio (UACR), patients were divided into pure DM group (group DM1, n=90), early diabetic nephropathy group (group DM2 , n=73) and clinical diabetic nephropathy group (group DM3 ,n=75). Clinic data of all patients were collected; Fasting blood glucose (FBG), 2h postprandial blood glucose (2hPB), blood lipids, uric acid (UA), fibrinogen (Fg) and glycosylated haemoglobin (HbA1c) were measured in all patients, and their correlations with DN were analyzed. Results: Compared with group DM1, the course of disease in DM [(7.25±6.29) years vs. (10.25±7.67) years vs. (13.53±7.82) years], levels of FBG [(8.46±2.52) mmol/L vs. (9.52±3.38) mmol/L vs. (10.82±3.30) mmol/L], 2hPB [(18.40±5.64) mmol/L vs. (20.27±5.94) mmol/L vs. (22.59±6.14) mmol/L], HbA1c [(7.96±1.65) % vs. (8.60±1.76) % vs. (9.55±2.09) %], triglyceride [TG, (1.72±0.86) mmol/L vs. (2.34±1.87) mmol/L vs. (3.16±1.85) mmol/L], Fg [(3.49±0.93) g/L vs. (3.88±1.21) g/L vs. (4.99±2.10) g/L] and UA [(295.42±52.34) μmol/L vs. (324.18±96.29) μmol/L vs. (351.23±56.88) μmol/L] significantly increased in group DM2 and group DM3 in order (P<0.01~0.001). Logistic gradual regression analysis indicated that course of DM, HbA1c, TG, Fg and UA were risk factors for DN (OR=1.008~1.910, P<0.01~0.001). Conclusion: The course of DM, blood glucose, blood lipid, uric acid and fibrinogen are risk factors of diabetic nephropathy; increased UACR reflects progress of patient’ condition in DM patients, its detection is used for diabetic prognosis and treatment.