Effect of endotoxin pretreatment-induced glycogen synthase kinase-3 inhibition on glycogen metabolism in rat liver and the mechanism / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the changes in the functional activity of glycogen synthase kinase-3 (GSK-3) in the hepatic tissue after endotoxin (lipopolysaccharide, LPS) tolerance and explore the effects of LPS-induced GSK-3 inhibition on glycogen metabolism in the liver.</p><p><b>METHODS</b>Male SD rats were randomly divided into normal control, endotoxin pretreatment and GSK-3 inhibitor (lithium chloride) groups with corresponding pretreatments prior to a large dose of LPS challenge (10 mg/kg) to induce liver injury. Glycogen deposition and content in the hepatic tissue was detected using periodic acid-Schiff (PAS) staining and a glycogen quantification kit, respectively. Western blotting was performed for semi-quantitative analysis of protein level and inhibitory phosphorylation of GSK-3, and a Coomassie brilliant blue G-250-based colorimetric assay was used to detect calpain activity in the liver.</p><p><b>RESULTS</b>Glycogen content in the liver decreased significantly after LPS challenge in all the 3 groups (P<0.05) but showed no significant difference among the groups (P>0.05). Both LPS and lithium chloride pretreatments caused a significant increase of liver glycogen content (P<0.05). LPS pretreatment induced inhibitory phosphorylation of GSK-3β (P<0.05) and partial cleavage of GSK-3α but did not affect the expression of GSK-3 protein (P>0.05). Large-dose LPS challenge significantly increased the activity of calpain in the liver tissue (P<0.05) to a comparable level in the 3 groups (P>0.05).</p><p><b>CONCLUSION</b>Endotoxin pretreatment induces inhibitory phosphorylation of GSK-3β and partial cleavage of GSK-3α and promotes the deposition of liver glycogen but does not affect the activity of calpain, which may contribute to an increased glycogen reserve for energy supply in the event of large-dose LPS challenge.</p>

Factors influencing the metastasis of Rotter's lymph node and its surgical dissection in patients with breast cancer / 中国肿瘤临床

Objective:This study aims to analyze the occurrence rate, positive rate, and other related factors influencing interpec-toral lymph nodes (IPNs) in breast cancer patients. This work further aims to explore the significance and indications of the surgical dis-section of IPNs. Methods:Clinical and pathological data from 1673 breast cancer patients were retrospectively analyzed. All patients were subjected to modified radical mastectomy, and IPNs were pathologically examined. The occurrence rate and metastasis of IPNs were recorded, and the relationship between the IPN positive rate and tumor size, axillary nodes, clinical stages, neo-adjuvant chemo-therapy, hormone receptors, Her-2 expression, and molecular subtypes of breast carcinoma was determined. Results:The occurrence rate, overall metastasis rate, and the positive rate of IPNs in patients with axillary lymph node metastasis were 13.39%, 4.30%, and 10.01%, respectively. IPN metastasis was significantly correlated with axillary node metastasis and the tumor, node and metastasis (TNM) stage of tumors (P<0.05). However, IPN metastasis was not significantly related with hormone receptor and Her-2 expressions. IPN metastasis rate may be unaffected by neo-adjuvant chemotherapy. Patients with IPNs metastasis were characterized by larger tu-mors, more positive axillary lymph nodes, and later TNM stages. Conclusion:IPN metastasis usually occurs in patients with larger tu-mors, more positive axillary lymph nodes, later TNM stages, as well as those with locally advanced cancer that meet the standard of neo-adjuvant chemotherapy. These indications suggest that the surgical dissection and pathological examination of IPNs should be rou-tinely performed.

Breast Conserving Surgery in Patients with Early Breast Cancer After Local Lumpectomy / 中国微创外科杂志

Objective To explore the feasibility and safety of breast conserving surgery ( BCS) in patients with early breast cancer after local lumpectomy . Methods Clinical data of 26 patients who previously had received local lumpectomy from January 2009 to December 2011 were retrospectively analyzed .All the patients were diagnosed as solitary invasive ductal carcinoma with preoperative staging of T1N0M0.The interval from lumpectomy to BCS was 6-24 days (mean, 10 days) and the maximum diameter of tumors before first operation was 1-2 cm (mean, 1.6 cm).The shortest distance between nipple and operative incision was 2-6 cm (mean, 4.5 cm). Results The operations were successfully completed in all 26 patients, 15 of which received sentinel lymph node biopsy and 11 of which received axillary lymph node dissection .The hospital stay was 3-5 days with stage-Ⅰhealing of incision .All patients received whole breast radiation therapy after BCS .There was no local recurrence or distant metastasis during 2 -5 year ’ s follow-up (median, 32 months).All the patients were satisfied with the shape of breast and the quality of life . Conclusion For those patients with early breast cancer who have received local lumpectomy , meet the requirement of CBS , and have the willing of breast conserving , BCS is an ideal choice after rigid application of surgical indications .

Expression of Toll-interacting protein (Tollip) in appendix during acute appendicitis / 第二军医大学学报

Objective To observe the expression of Toll-interacting protein ( Tollip) in appendix and analyze its significance during acute inflammation. Methods Thirty-three patients with acute appendicitis were included in the present study and 6 subjects with non-inflammatory appendixes were taken as controls ( without inflammatory changes). The pulse rate,body temperature ( BT) ,white blood cell ( WBC) count and neutrophil ( NEUT) count were observed one hour before appendectomy. Based on H-E staining and pathological examination,the appendix samples were divided into four groups:non-inflammatory appendix ( A) ,simple appendicitis ( B) ,suppurative appendicitis ( C) and gangrenous appendicitis ( D). The expression of Tollip protein ( the localization,qualitative and semiquantitative analysis) was analyzed using immunohistochemistry and digital image analysis. The correlation of Tollip with pusle rate,BT,WBC and NEUT was also analyzed. Results Significant differences in BT,WBC and NEUT were found between different pathological groups ( P

THE EXPRESSION OF HUMAN MD-2/GST FUSION PROTEIN IN E. coli / 解放军医学杂志

To design and construct an expression vector,PGEX-4T-1/MD-2, and to express human myeloid differentiation protein-2(hMD-2) and glutathione-S-transferase (GST) fusion protein in E. coli, the EcoRI/SalI sites and stop code were incorporated into the hMD-2 encoding fragment by PCR. After digesting with EcoRI/SalI, the hMD-2 encoding fragment was cloned into the expression vector PGEX-4T-1 at the corresponding sites. The positive clones selected with PCR and restriction endonuclease digestion were sequenced and the expression of GST/hMD-2 fusion protein in E. coli BL21 was analyzed with SDS-PAGE after induced by 0.4mmol/L IPTG for 3 to 5 hours. The hMD-2 encoding fragment containing stop code was correctly inserted into the expression vector PGEX-4T-1 and this was confirmed by PCR, double-enzyme identification and sequencing. The SDS-PAGE analysis indicated that the GST/hMD-2 fusion protein was successfully expressed in E. coli and the yield of the fusion protein was 30 percent of bacterial total protein. The construction of the expression vector PGEX-4T-1/hMD-2 and the expression of fusion protein GST/hMD-2 in E. coli would be useful for further investigation of MD-2.