Objective To explore the feasibility and safety of day surgery laparoscopic cholecystectomy . Methods From January 2015 to June 2015, 78 cases of day-case laparoscopic cholecystectomy were performed in our hospital .The clinical data were retrospectively reviewed .These patients were diagnosed as having cholelithiasis or cholecystic poypus .They had no severe heart or lung diseases, no history of epigastric operation , and agreed to accept day-case laparoscopic cholecystectomy without overnight hospitalization. Results Of the 78 patients, 74 patients (94.9%) successfully accepted the surgery and were discharged no more than 4 h after surgery, including 2 patients with combined laparoscopic transcyctic common bile duct exploration ( LTCBDE).The remaining 4 patients (5.1%) were admitted for overnight hospital stay , including 1 patient (1.3%) with Mirizzi syndrome type Ⅰwho was given abdominal drainage , 2 patients (2.6%) with severe nausea and vomit , 1 patient (1.3%) with airway obstruction after operation.All the 4 patients were discharged within 24 h after the surgery.The operating time was 35-152 min (mean, 80.0 ±19.7 min).The hospitalization expenses was 5688-9768 yuan (mean, 8318.0 ±848.5 yuan).The hospitalization time was 5.6-23.9 h (median, 7.4 h).Postoperative follow-up at the day of surgery, the first and the seventh postoperative day showed that none of the patients suffered from severe complications . Conclusion Day-case laparoscopic cholecystectomy is safe and feasible , on the basis of strictly selected patients , experienced surgeons , and close cooperation with anesthetists and nurses .

Actinomyces ; isolation & purification ; Actinomycosis ; microbiology ; Apicoectomy ; methods ; Candida albicans ; isolation & purification ; Candidiasis ; microbiology ; Enterococcus faecalis ; isolation & purification ; Foreign Bodies ; complications ; Gram-Positive Bacterial Infections ; microbiology ; Humans ; Microsurgery ; methods ; Periapical Periodontitis ; etiology ; microbiology ; surgery ; therapy ; Radicular Cyst ; complications ; Root Canal Filling Materials ; therapeutic use ; Root Canal Therapy ; methods

Objective To obtain recombinant human granulysin using prokaryotic expression system. Methods Total RNA was extracted from cultured PBMC. Granulysin gene segments were obtained with granulysin-specific primers by RT-PCR and then inserted into pET32a(+) plasmid. After identification by DNA sequence, pET-GN-LY9K and pET-GNLY15K were transferred to E. Coli Rosetta (DE3). The fusion protein was identified by SDS-PAGE and Western blot. The bioactivity of granulysin fusion protein was measured by MTT assay. Results The prokaryotic expression vectors pET-GNLY9K and pET-GNLY15K were successfully constructed. The corresponding protein was highly expressed in E. Coli. Recombinant protein was specifically bound by anti-granulysin antibody. GNLY9K fusion protein significantly inhibited the growth of A549 cells in a dose-dependent manner, while GN-LY15K had little effect on the growth of A549. Conclusion Granulysins with different mw were successfully expressed using prokaryotic expression system, which might be helpful for the further study of granulysin.

Objective To analyze clinicopathologic characteristics and prognostic factors of the patients with invasive lobular carcinoma of breast. Methods Clinical data of 125 patients with invasive lobular carcinoma of breast treated at Cancer Center of Sun Yat-sen University from Jan. 1990 to Dec. 2008, were analyzed. The clinical characteristics, recurrence and survival of the patients were summarized. Results Median age of 125 patients was 45 years old (range, 27 to 76 years old). The patients with large tumor mass (≥ 3cm), positive local lymph node, more than Ⅱ stage and positive hormone receptor at diagnosis were 77 cases(61.6 %), 64 cases(51.2 %), 101 cases(80.8 %) and 112 cases(89.6 %), respectively. The median time of follow-up was 58 months (range, 11-222 months). Of the 125 patients, 32 had local recurrence and metastasis, and 18 died. The 5-year disease-free and overall survival rates were 82.2 % and 87.3 %, respectively. Multivariate COX regression analysis showed that whether endocrine therapy or not was only a prognostic factor of patients with invasive lobular carcinoma of breast. Conclusion There is no difference in media age of patients with invasive lobular carcinoma of breast at diagnosis from other pathologic type of breast cancer. These patients are usually with larger tumor masses, more lymph node metastasis and a higher proportion of positive hormone receptor. The prognosis of patients is not affected by clinicopathologic parameters.

Annexin A3 ; genetics ; metabolism ; Cells, Cultured ; Guanine Nucleotide Dissociation Inhibitors ; genetics ; metabolism ; Hepatocytes ; drug effects ; metabolism ; Humans ; Lactoylglutathione Lyase ; genetics ; metabolism ; RNA, Messenger ; genetics ; Trichloroethylene ; toxicity ; rho-Specific Guanine Nucleotide Dissociation Inhibitors

Objective To obtain recombinant human granulysin using prokaryotic expression system.MethodsTotal RNA was extracted from cultured PBMC. Granulysin gene segments were obtained with granulysin-specific primers by RT-PCR and then inserted into pET32a(+) plasmid. After identification by DNA sequence,pET-GNLY9K and pET-GNLY15K were transferred to E. coli Rosetta (DE3). The fusion protein was identified by SDS-PAGE and Western blot.The bioactivity of granulysin fusion protein was measured by MTT assay.Results The prokaryotic expression vectors pET-GNLY9K and pET-GNLY15K were successfully constructed.The corresponding protein was highly expressed in E.coli. Recombinant protein was specifically bound by anti-granulysin antibody. GNLY9K fusion protein significantly inhibited the growth of A549 cells in a dose-dependent manner,while GNLY15K had little effect on the growth of A549.Conclusion Granulysins with different mw were successfully expressed using prokaryotic expression system,which might be helpful for the further study of granulysin.

Objective To study bioactive metabolite produced by marine actinomyces M326.Methods Based on the index of antimicrobial activity,the relation between activity of metabolite and conditions of culture in different media and time were researched,and the stability of bioactive metabolite treated with different temperatures and pH were obseved,and the bioactive ingredients were extracted and separated by organic solvent and by macroporous resin.Results The GBP medium made by artificial seawater or distilled water were suitable for production of strong bioactive metabolites.The antimicrobial substances were stable under pH 2～11 and were heat-resistant under strong acidic condition.It could not be extracted by usual organic solvent,but could be absorbed by AB-8 macroporous resin under alkaline situation.Conclusion The metabolites have strong activity against gram-positive bacteria and have different antimicrobial activities against gram-negative bacteria and drug-resistant strains moreover,the polarity of antimicrobial substances is strong.

Objective To investigate the short term effect of insulin on proinsulin gene expression of HIT-T15 insu-linnoma cells(pancreatic isletβ-cell). Methods The HIT-T15 cells were randomly divided into four groups.Blank Con-trol Group (LG):complete medium contain 1.4 mmol/L glucose. Control group (LGC):co-cultured nifedipine with medium in order to restain endogenous insulin release. Experimental group (LINS or HINS) add 0.5 U/L insulin or 5 U/L insulin on top of LGC. After being stimulated for 0, 30, 60, 90, 120 mins, proinsulin (PI) mRNA level were assessed by semi-quantitative RT-PCR. Insulin receptor substrate1 (IRS1) tyrosine phosphorylation was detected by immunocytochemistry. Results (1) Expression of PI was up regulated by both LINS and HINS, and peak at 60 mins. (2) After stimulation for 30 mins, the level of IRS1 tyrosine phosphorylation in the experimental group was significantly higher than control group, and the peak time be-tween LINS and HINS was different. (3) Between group of LG and LGC, the expression of PI mRNA and IRS1 tyrosine phos-phorylation show no difference. Conclusion Short term exogenous insulin stimulation can promote expression of proinsulin genes,which is concentration dependent. The expression and regulation of PI were related with IRS1 signal transduction.

Objective To investigate the expression of miR-29s in the glioma stem cells,and explore how the members of miR-29s affect the bio-logical behaviors of glioma stem cells. Methods Eight patient specimens were used to culture glioma stem cells. Real-time PCR was adopted to test the expression of miR-29s. CCK-8 analysis was performed to test the proliferation ,Transwell was used to test cell migration and invasion ,and flow-cytometry analysis was carried out to test apoptosis. Results miR-29a,miR-29b and miR-29c were decreased in glioma stem cells. Over-ex-pression of miR-29s could inhibit the proliferation,cell migration and invasion,but promote apoptosis of glioma stem cells. Conclusion miR-29s acts as a cancer suppressor gene in the glioma stem cells ,and miR-29a plays the dominant functional role in the family.

Objective To observe effect of environmental enrichment on the learning and memory ability and the expressions of hypoxia inducible factor 1α (HIF-1a) and vascular endothelial growth factor (VEGF) in rats with chronic cerebral hypoperfusion.Methods A total of 40 rats were randomly divided into 3 groups: bilateral vascular occlusion (2VO) of the common carotid arteries group (n=14, 2VO group), 2VO + enriched environment (EE) group (n=14, 2VO+EE group) and sham group (n=12, SHAM group).Morris water maze, novel object recognition test, real-time reverse transcription-polymerase chain reaction (real-time RT-PCR), immunohistochemistry methods and Western blotting were used to detect changes in learning and memory ability of rats and HIF-1α and VEGF expression levels in hippocampus.Results Morris water maze showed that the escape latency was longer in the 2VO group than in the SHAM group at 3, 4 and 5 day during the training (all P＜0.05), while the 2VO+EE group spent significantly less time in finding the platform as compared with the 2VO group at 4 and 5 day (both P＜0.05).The time for space exploration in target quadrant was less in 2VO group than in SHAM group (P＜0.05), while it was longer in 2VO +EE group than in 2VO group (P＜0.05).Novel object recognition test showed that the 2VO operation impaired the priority index (PI) of time spending at exploring the novel object (P＜0.05), and environmental enrichment could improve the PI in 2VO group (P＜0.05).The real-time RT-PCR and Western blotting showed that the HIF-1α mRNA expression was higher in 2VO group than in SHAM group (P＜0.05).The VEGF mRNA and protein expressions were higher in 2VO+EE group than in 2VO group (both P＜0.05).The expression of HIF-1α in hippocampal CA1 area was higher in 2VO group than in SHAM group (P＜0.05).Conclusions Environmental enrichment can alleviate the damages of spatial and non-spatial learning and memory ability which are caused by chronic cerebral hypoperfusion.And HIF-1α and its downstream gene VEGF may be involved in the restoration of cognitive function by enriched environment.