Objective:To investigate the expression of PTEN(phosphatase and tensin hmmlogy deleted on ten) in the breast cancer and the relationship with the tumor angiogenesis.Methods:The expression of PTEN and MVD were detected by immunohistochemical SP method in paraffin section from 53 women with primary breast cancer and 10 subjects of hyperplasia of mammary glands.Results:The higher lever PTEN expression in 53 case with the rate 49.1 %(26/53),which was reduced compared with the expression in benign hyperplasia of mammary glands(P

OBJECTIVE To investigate the mechanisms for carbapenem resistance in clinical isolates of Pseudomonas aeruginosa from surgical intensive care unit(SICU).METHODS The minimal inhibitory concentrations(MICs) of various antibiotics were detected with agar dilution method;their ?-lactamases were extracted,types of their enzymes were identified and then their physical and chemical characteristics were studied by three dimentional extract test and isoelectric focusing(IEF) electropheresis;primers specific to IMP and VIM genes were utilized to amplify the metallo-?-lactamase genes.The levels of outer membrane protein OprD2 were measured by Western blot.RESULTS Forty one strains were resistant to imipenem from 49 strains of P.aeruginosa isolated from SICU during 3 years.Among them,34 isolates were found to produce high level of AmpC enzymes.Eight produced extended-spectrum ?-lactamases(ESBLs) at the same time.2 produced ESBLs only.No metallo-?-lactamases were detected.The activity of enzymes in the imipenem resistant group(74.32?53.42) was statistically different from the sensitive one(8.7?16.16,P

Objective To investigate the mechanism of the permeability of intestinal mucosa in the pathogenesis of irritable bowel syndrome (IBS) and the interventional effects of Clostridium butyricum combined with glutamine.Methods According to random number method,fifty BALB/c mice were divided into control group,experimental control group,glutamine group,Clostridium butyricum group and combination group.IBS mice model was established by water-avoidance stress (WAS) experiment.The defecating time of mice and fecal water content were detected by dyed stool after mice gavaged with methylcellulose (1.5％).The pathological injury of intestine was assessed by hematoxylin and eosin staining.The visceral sensitivity was evaluated by colorectal distention test (CRD).The changes of the permeability of intestine was evaluated by detecting the changes of serum D-lactic acid (D-LA),level of diamine oxidase (DAO),expressions of intestinal epithelial cells (IEC) cell tight junction protein (TJ) (occludin-1,claudin-1,zonula occludens-1 (ZOL-1)) at protein level.The interventional effects of Clostridium butyricum combined with glutamine were evaluated.t test was performed for comparison between groups,and analysis of variance was used for comparison among multi-groups.Results Compared with the control group,the defecating time of experimental control group was significantly shorten ((100.40±14.80) min vs (75.88±12.20) min and water content of fecal significantly increased ((54.76±9.98)％ vs (74.95±7.15)％,t =3.692 and 4.023; P=0.002 and 0.002).The lowest threshold of visceral sensitivity significantly decreased ((40.87 ± 4.82) mmHg (1 mmHg=0.133 kPa) vs (27.80±3.18) mmHg; t=8.761,P＜0.01),while the mucosal pathological injury score significantly increased (0.50±0.15 vs2.60±0.97; t＝6.034,P＜0.01).The level of D-LA ((1 476±246.8) ng/L vs (913.6±90.1) ng/L)) and DAO ((3 391.0±256.9) vs (5 096.0±725.2) ng/L) significantly increased (t=40.920 and 29.810; both P＜0.05),and the expression of tight junction protein ZOL-1 (0.165±0.005 vs0.119±0.003),occludin-1 (0.104±0.016 vs 0.022±0.006) significantly decreased (t=19.830 and 19.830; both P＜0.01).Compared with the experimental control group,after intragastric intervention the defecating time of glutamine group,Clostridium butyricum group and combination group increased ((90.50±3.78),(97.56±8.79) and (99.89±11.90) min and water content of fecal decreased ((69.33±6.71)％,(58.07±8.97)％ and (56.74±8.12)％) and the differences were statistically significant (F=10.020 and 8.740; both P＜0.01).The results of Clostridium butyricum group and combination group were good (F=2.481 and 4.874; both P＜0.05).And the lowest threshold of visceral sensitivity significantly increased ((31.80±2.69),(36.04±5.06) and (38.93±3.30) mmHg; F=2.420,P＜0.05),the result of combination group was the best (F=3.550,P＜0.01).Jejunal mucosal injury was significantly reduced (2.00 ± 0.94,1.30 ± 0.68 and 1.30±0.48; F＝11.350,P＜0.01).After intragastric intervention,serum levels of D-LA ((1 370.0± 78.9),(1 066.0±155.5) and (1 039.0±129.0) ng/L) and DAO ((4 808.0±477.4),(3 713.0± 595.0) and (3 725.0±615.9) ng/L) of glutamine group,Clostridium butyricum group and combination group significantly decreased (F＝37.480 and 27.670; both P＜0.01).The level of ZOL-1(0.126± 0.014,0.125±0.006,0.138±0.004) and occludin 1 (0.037±0.013,0.073±0.028,0.078±0.027) of glutamine group,Clostridium butyricum group and combination group significantly increased,and the differences were statistically significant (F=5.867 and 10.630; both P＜0.05).The change of ZOL-1 of combination group was more than that of Clostridium butyricum group (t =5.457,P ＜ 0.05).Conclusions WAS experiment can induce visceral hypersensitivity,increase the permeability of intestine and reduce the function of intestinal epithelial barrier.Clostridium butyricum and glutamine are effective in the recovery of visceral hypersensitivity and the permeability of mucosal epithelia cells.

Objective To investigate the effects of Bifidobacterium infatn ison the expression of in -testinal corticotropin releasing factor ( CRF) receptors and how the peripheral CRF receptors activate mast cells in a murine model of irritable bowel syndrome (IBS).Methods Thirty BALB/c male mice were ran-domly divided into three groups including control group , model group and Bifidobacterium infantis group. The mouse model of IBS was established by using chronic restraint stress .Mice in Bifidobacterium infantis group received daily intragastrical administration of Bifidobacterium infantis for 14 days.Mice in control and model groups were treated with equal volume of saline .Then all mice were killed after the assessment of weight and abdominal withdrawal reflex ( AWR) .The levels of histamine , tryptase and tumor necrosis fac-tor-α( TNF-α) in serum samples were detected by ELISA .The expression of CRF in colonic mucosa was analyzed by immunohistochemistry .The expression of CRF-R1 and CRF-R2 in mast cells and the number of mast cells in colonic mucosa were detected by double immunofluorescence staining assay .The expression of CRF-R1 and CRF-R2 at mRNA level in colon were detected by reverse transcription polymerase chain reac-tion ( RT-PCR) .Results Compared with control group , the levels of histamine , tryptase and TNF-αin pe-ripheral blood samples , the expression of CRF-R1 and CRF-R2 at mRNA level , and the number of mast cells, CRF-R1+mast cells and CRF-R2+mast cells in colonic mucosa were increased significantly in model group (P<0.05), but were remarkably down-regulated with the treatment ofB ifidobacterium infantis (P<0.05).Conclusion Bifidobacterium infantis could reduce the activation of mast cells in a murine model of IBS by inhibiting the expression of CRF-R1 and CRF-R2 in intestinal mast cells .

Objective To investigate the effect of corticotrophin-releasing factor (CRF) on the regulation of Toll-like receptor 4/NF-κB signaling pathway expression in human intestinal epithelial cell line HT-29. Methods HT-29 cells were divided into four groups, normal control group, LPS group (LPS 20 μg/ml stimulated for 24 h), CRF group (CRF 20 ng/ml stimulated 24 h) and CRF+ LPS group (CRF incubated for 12 h then changed to LPS for another 12 h). After stimulation, the expression of TLR4 mRNA of each group was examined by reverse transcriptase polymerase chain reaction (RT-PCR). Total cell protein were extracted and the expression of TLR4 and NFκB p65 at protein level were detected by western blotting.Cell culture supernatant was collected and the secretion of interleukin-8 was detected by enzyme-linked immunoasorbent assay (ELISA). Results The expression of TLR4 in LPS group at mRNA and protein level were 0.31±0.04 and 0.48±0.17,there was no significant difference compared with normal control group (0.28±0.02 and 0.45±0.12,t＝0.216 and 0.712 , P＞0.05 ). In CRF group which were 1.05±0.06 and1. 08±0.21, significantly higher than normal control group (t＝3.721 and 3.802, P＜0.05). In CRF+LPS group which were 1.68±0.05 and 1.81±0. 18,significantly higher than CRF group (t＝4. 816 and 3. 918, P＜0.05).The results of NF-κB p65 expression at protein level and interleukin-8 expression of cell culture supernatant were consistent with the results of TLR4 expression at mRNA and protein level.Conclusion CRF not only activate TLR4/NF-κB signaling pathway in human intestinal epithelial cell,but enhance the reaction of intestinal epithelial cell to LPS as well, which resulting in increased interleukin-8 secretion.

Objective To study the presentation,sensitivity of different diagnostic methods,snrgical modalities and pathological findings for patients of substemal goiter. Methods The clinical data of fifty nine cases of substernal goiter surgically treated were retrospectively analyzed. Results The main symptoms of substernal goiter patients were asymptomatic cervical mass(39/59),airway obstruction (13/59),hyperthyroidism(4/59),hoarseness(3/59)and choke(3/59).The sensitivity of chest X-ray,ultrasonography,CT and scintigraphy was 62.8%,15.8%,85.7%,and 50.0%respectively,and the specificity Was 99.4%,99.8%,99.5%and 99.0%,respectively.Standard cervical approach was successful for thyroidectomy operations in 57 cases.One patient underwent thyroidectomy by sternotomic approach,and another one by combined cervici-sternotomic approach.No major complications or perioperative deaths occurred.Pathology revealed nodular goiter in 48 cases.Grave's disease in one,thyroid adenoma in 2,and carcinoma in 8. Conclusions CT and chest X-ray are sensitive diagnostic techniques for substerual goiter.Cervical approach is appropriate for most substernal goiter.

Objective To investigate the role of intestinal immune dysfunction in the pathogenesis of irritable bowel syndrome(IBS) and to study the effects of Clostridium butylicum on the regulation of intestinal immune disorders.Methods A total of 50 male 6-week-old C57BL/6 mice were randomly divided into three groups,including the experimental group (n =20),the control group (n =20) and the Clostridium butylicum group(n =10).A mouse model of constipation-predominant IBS (C-IBS) was established by perfusing sodium butyrate solution(200 μl,concentration of 500 mmol/L) into the mouse colon twice a day for three consecutive days.The mice in control group were intrarectally perfused with normal saline enema (200 μl).Two hours before the perfusion of sodium butyrate into colon,the mice in Clostridium butylicum group were given Clostridium butylicum 500 μl(viable cell concentration of 1×109 CFU/ml) by oral gavage once a day for six days.The colorectal distention test(CRD) was carried out for evaluation of clinical parameters.HE staining of intestinal tissue section was performed for histopathological assessment of colonic mucosal inflammation.Intestinal intraepithelial lymphocytes (IELs) and lamina propria mononuclear cells (LPMCs) were isolated and analyzed by flow cytometry to evaluate the correlation between IBS and intestinal immune dysfunction/abnormal activation of intestinal immune cells in mouse model of C-IBS,and to assess the regulatory effects of Clostridium butylicum on the intestinal immune disorder.Results (1) Compared with the control group,the mice in experimental group showed a significant change in physiological parameters,histological structure of colon,inflammatory cells infiltration and low-grade inflammatory state.There was a significant increase in scores of CRD and a decrease in lowest sensory threshold (t=8.926 and t=6.103,both P＜0.001) ; (2) There was a decrease in the numbers of DC in IELs (t =2.878 and t =3.086,both P＜0.05),but an increase in the numbers of macrophage (t=3.191,P＜0.05) and the memory T cells in mice with IBS (t=3.071,P＜0.05) as compared with that in control group; (3)DCs were decreased (t=2.880 and t=2.664,both P＜0.05),but memory T cells were increased (t =3.732 and 2.682,P＜0.01 and P＜0.05) in the LPMCs of mice in experimental group; (4)There was no significant difference in the physiological index between the mice in control group and the Clostridium butylicum group.Levels of memory T cells,macrophages and DCs in the IELs were close to the normal level (6 d,t =1.103,0.0213,0.418,all P＞0.05),and levels of macrophages and DCs in the LPMCs of mice in the Clostridium butylicum group were also similar to that in the control group (6 d,t =0.782,0.347,both P＞0.05) ; (5) Compared with the mice in experimental group,the level of memory T cells in LPMCs of mice treated with Clostridium butylicum was dramatically declined (6 d,t=2.346,P=0.0470,P＜0.05),however,which was still higher than that of mice in control group (6 d,t =2.233,P =0.0476,P＜0.05).The intestinal immune function was restored to normal level with Clostridium butylicum intervention.Conclusion The pathophysiologic mechanism of IBS might be closely related to the abnormal activation of intestinal immune cellsand disordered functional state in the intestinal mucosa.Clostridium butylicum could regulate the intestinal immune homeostasis and restore the physiological function of gastrointestinal tract.

Objective To explore the accuracy and feasibility of dual-source CT(DSCT)in evaluating the left ventricular function compared with echocardiography.Methods 30 cases of coronary heart disease were continuously collected in our hospital ,and dual-source CT and echocardiography were examined in a week.Left ventricular function parameters (LVEDV,LVESV,LVSV)were measured,and paired sample t test was compared to show whether there was any difference between the two methods,Rates betweengroups were compared with Chi-square test,Pearson correlation analysis method was applied to calculate the correlation between the two methods.Results There was no statistically significant difference for LVEDV,LVESV,LVSV,LVEF parameters between the two methods (t test,P >0.05).By Pearson test,the correlation coefficient was r=0.770-0.938(P <0.05),in which the high relation between the two methods was showed.Conclusion A strong accuracy of dual-source CT in evaluating left ventricular function is demonstrated.Dual-source CT angiography is a noninvasive assessment of coronary tree,whereas the analysis of the left ventricular parameters provides additional information of cardiac function without further radiation exposure or scan time,which can provide a more intuitive and system information for clinical.

A glycomic method was used to screen the aberrantly ?1-6 fucosylated glycoproteins related to HCC metastasis and analyze the alteration of CK8 both in its expression level and its glycan parts associated with metastatic ability. Based on the approach, 2-DE coupled with lectin affinity blot, lectin affinity precipitation followed by MALDI-TOF-MS/MS, the lens culinaris agglutinin (LCA) affinity glycoprotein profiles from MHCC97-L and MHCC97-H cells, two higher metastatic HCC cell lines, were obtained, in which a differentially displayed protein spot was indicated when compared with Hep3B, in the region within 55～60 ku in molecular mass and 4～6 in isoelectric point. The identification result was CK8 by MALDI-TOF-MS/MS. To confirm the relation between increased core-fucosylation of CK8 and HCC metastasis, LCA affinity precipitation was used to extract the ?1-6 fucosylated glycoproteins, followed by Western blot. And it was found that CK8 was highly fucosylated in both MHCC97-L and MHCC97-H cells compared to Hep3B. Immunofluorescence analysis and Western blot were used to detect its intracellular localization and its protein expression levels, indicating that CK8 distributed in cytoplasm and increased protein expressions in MHCC97-L and MHCC97-H cell lines. And further lectin binding studies found that CK8 has a high affinity for Con A in both MHCC97-H and Hep3B cells, indicating that CK8 was a glycoprotein with high-mannose type N-glycans. But the amount of the lectin RCA-1 binding to CK8 was greater in MHCC97-H than Hep3B, suggesting that CK8 contained the increased terminal galactose residues ?-1, 4-linked to GlcNAc in MHCC97-H. All the results suggested that the increase of CK8 in its protein expression level, core-fucosylation and terminal gal ?1,4 GlcNAc disaccharides might be related to HCC metastatic ability.

Objective SD rats were immuned with the transgenic tomatoes which carried fused gene of a region of PAc Strep-tococcus mutants and cholera toxin B subunit.The immunogenicity was tested to explore secure and economic edible vaccines a-gainst dental caries.Methods A total of 18 eighteen-day-old female SD rats were subdivided randomly into three groups:the exper-imental group which were fed with transgenic tomato juice containing chimaera protein PAcP/CTB;the positive control group which were treated with deactivated S.mutans;the negative control group which were not treated with transgenic tomato juice.Rats were immuned once per week for four weeks.Blood and saliva were collected at one day before the first immunity and one week after each immunization.IgG of blood serum and SIgA of saliva were detected using the enzyme-linked immunosorbent assay (indirect ELISA) testing.On day 70,rats were terminated.The maxillary and mandibular bones were subsequently taken out to count dental caries′scores.Results Post immunization,the experimental group and the positive control group had statistical significant levels of speci-ficity IgG in serum and SIgA in saliva compared to the negative control group (P <0.05).There was a significance difference be-tween the experimental group and the negative control group except in Dx levels of caries loss (P <0.05).Conclusion The targeted protein expressed on the transgenic tomatoes is immunogenic,which can effectively induce mucous membrane immune response and the systematical immunoreaction to suppress the occurrence of the dental caries.