OBJECTIVE To explore the role of gecko crude peptides (GCPs) in the proliferation, apoptosis, migration and lymphangiogenesis of human hepatocellular carcinoma cells (HepG2) and human lymphaticendothelial cells (HLECs) in vitro. METHODS The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to evaluate the anti- proliferative effect of GCPs and siRNA-VEGF-C on HepG2 cells, Hoechst 33258 staining and flow cytometry were performed to analyze cycle and apoptosis. The migration and invasion ability of cells were assayed by transwell chamber experiment and wound-healing assay. The protein and mRNA expressions of vascular endo?thelial growth factor-C (VEGF-C) and CXC chemokine receptor-4 (CXCR4) were detected by q-PCR, immunofluorescence, Western blot. The protein expressions of the extracellular signal regulated kinase (ERKI/2), c-Jun N-terminal kinase (JNK), p38-mitogen activated protein kinases (p38 MAPK), serine/threonine kinase (Akt) and phosphatidylinositol- 3- kinase (PI3K) were detected by western blot. The anti-lymphangiogenesis effect of GCPs on the HLECs was analyzed using an in vitro tube-formation assay. The protein and mRNA expressions of vascular endothelial growth factor receptor-3 (VEGFR-3) and stromal cell-derived factor-1 (SDF-1) were detected by q-PCR, Western blot. RESULTS GCPs and siRNA-VEGF-C inhibited HepG2 proliferation, invasion and migration, and the most obvious inhibitory effect was both synergistic effects. Thus, GCPs suppressed HLECs proliferation, migration and tube-like structure formationin a dose- dependent manner, and had inhibitory effect of tumor- induced lymphangiogenesis in vitro. Additionally, we found that GCPs and siRNA- VEGF- C decreased the expressions of MMP-2, MMP-9, VEGF-C, CXCR4, phospho-ERK1/2, phospho-P38, phospho-JNK and PI3K in HepG2 cells. Moreover, GCPs had a dose-dependent depressive effecton the expressions of VEGFR- 3, SDF- 1 in HLECs. CONCLUSION The low expression of VEGF- C mediated by siRNA-VEGF-C and GCPs inhibit tumor proliferation, invasion and migrationby suppressing the MAPK signaling pathway through reduced levels of VEGF-C, and GCPs inhibit tumor lymphangiogenesis by suppressing the CXCR4/SDF-1 signaling pathway through suppressed VEGF-C/VEGFR-3.

Radiotracer techniques were employed to characterize 65Zn adsorption and desorption in root-cell-wall of hyperaccumulating ecotype (HE) and non-hyperaccumulating ecotype (NHE) species of Sedum alfredii Hance. The results indicated that at the end ofa 30 min short time radioisotope loading period, comparable amounts of 65Zn were accumulated in the roots of the two ecotypes Sedum alfredii, whereas 2.1-fold more 65Zn remains in NHE root after 45-min desorption. At the end of 60 min uptake period, no difference of 65Zn accumulation was observed in undesorbed root-cell-wall of Sedum alfredii. However, 3.0-fold more 65Zn accumulated in desorbed root-cell-wall ofNHE. Zn2+ binding in root-cell-wall preparations of NHE was greater than that in HE under high Zn2+ concentration. All these results suggested that root-cell-wall of the two ecotypes Sedum alfredii had the same ability to adsorb Zn2+, whereas the desorption characteristics were different, and with most of 65Zn binding on root of HE being available for loading into the xylem, as a result, more 65Zn was translocated to the shoot.

Animals ; CHO Cells ; Cricetulus ; Doxycycline ; pharmacology ; Immunohistochemistry ; Ki-67 Antigen ; genetics ; metabolism ; Plasmids ; Quality Control ; Staining and Labeling ; methods ; Transfection

OBJECTIVETo investigate c-myc and CCNE2 gene amplifications and their relationship in breast cancer.

METHODSSixty-six infiltrating ductal breast carcinomas with foci of ductal carcinoma in situ components collected from January 2005 to December 2007 were selected for tissue microarray and quantitative multi-gene FISH for c-myc and CCNE2 gene amplification, and the relationship with the clinicopathologic features was analyzed.

RESULTSOf the 66 cases, 18 (27.3%) showed c-myc amplification and 23 (34.8%) showed CCNE2 amplification. A strong correlation was found between c-myc and CCNE2 amplification (P < 0.01). The breast cancers showing c-myc and CCNE2 amplifications were all aneuploidy, and were HER2 positive (P < 0.05). Tumors with c-myc amplification also showed higher Ki-67 index (P < 0.05).

CONCLUSIONSC-myc and CCNE2 amplifications are common events in breast cancer, and they often coexist. C-myc and CCNE2 genes may play critical roles in the pathogenesis and development of breast cancer through unique and overlapping signaling pathways.

Aneuploidy ; Breast Neoplasms ; genetics ; Carcinoma in Situ ; genetics ; Carcinoma, Ductal, Breast ; genetics ; Carcinoma, Intraductal, Noninfiltrating ; genetics ; Cyclins ; genetics ; Female ; Gene Amplification ; Genes, myc ; Humans ; In Situ Hybridization, Fluorescence ; methods ; Tissue Array Analysis

OBJECTIVETo study the effect of rutin on body weight and obesity-induced reproductive impairment in male mice.

METHODSTwenty-four male mice were randomized equally into normal control group, high-fat diet group (HFD group), and HFD + rutin intervention group (HRU group). After 28 days of treatments, the testes and epididymis of the mice were collected for detection of total cholesterol (TC) and triglycerides (TG) levels and for pathological examinations with HE staining. The expressions of related genes was detected with real-time PCR, and Western blotting was used to detect the expression of Ucp1 protein in the samples.

RESULTSAfter 28 days of treatments, the mean body weight was lower in mice with rutin intervention than in those in HFD group. The mice in HFD group showed significantly higher TG levels in the testis and epididymis and higher TC levels in the epididymis than those in the control and HRU groups. In HFD group, the testis and the epididymis displayed loosened structures with abnormalcell structure, and the number ofmature spermatozoa in the lumen was decreased and the mobility of the sperms was reduced; all these changes were significantly alleviated in HRU group. The expression levels of Ucp1 mRNA and protein increased (P<0.05) and the expressions of Mcp1 and TNF-α decreased significantly in the mice after rutin treatment (P<0.05).

CONCLUSIONRutin can effectively inhibit rapid increase of body weight and protect against obesity-induced reproductive impairment in obese mice.

OBJECTIVETo assess the prognosis and reproductive outcomes of laparoscopic intracapsular myomectomy.

METHODSA total of 673 women received subserosal and intramural intracapsular laparoscopic myomectomy between March, 2007 and March, 2012, and their post-operative complications, the need for subsequent surgery, symptomatic relief and reproductive outcomes were analyzed.

RESULTSOf these patients, 42.4% had subserosal myomas and 57.6% had intramural myomas. The mean total operative time was 96∓41 min with a mean blood loss of 128∓46.2 ml, and 82.3% of the patients were discharged 48 h after the operation without early complications. A small fraction (2.3%) of the patients had a second laparoscopic myomectomy for recurrent fibroids. Of the fertility-demanding women who underwent myomectomy, 71% achieved pregnancy, 49.8% underwent caesarean section, 8% had operative vaginal deliveries, and 42.2% had spontaneous deliveries; uterine rupture occurred in none of the cases.

CONCLUSIONLaparoscopic intracapsular myomectomy, by preserving the fibroid pseudocapsule and myometrial integrity, has no early postoperative complications and ensures good fertility rates and reproductive outcomes.

Adult ; Female ; Fertility ; Humans ; Laparoscopy ; Leiomyoma ; surgery ; Prognosis ; Retrospective Studies ; Uterine Myomectomy ; Uterine Neoplasms ; surgery

Radiotracer techniques were employed to characterize (65)Zn adsorption and desorption in root-cell-wall of hyperaccumulating ecotype (HE) and non-hyperaccumulating ecotype (NHE) species of Sedum alfredii Hance. The results indicated that at the end of a 30 min short time radioisotope loading period, comparable amounts of (65)Zn were accumulated in the roots of the two ecotypes Sedum alfredii, whereas 2.1-fold more (65)Zn remains in NHE root after 45-min desorption. At the end of 60 min uptake period, no difference of (65)Zn accumulation was observed in undesorbed root-cell-wall of Sedum alfredii. However, 3.0-fold more (65)Zn accumulated in desorbed root-cell-wall of NHE. Zn(2+) binding in root-cell-wall preparations of NHE was greater than that in HE under high Zn(2+) concentration. All these results suggested that root-cell-wall of the two ecotypes Sedum alfredii had the same ability to adsorb Zn(2+), whereas the desorption characteristics were different, and with most of (65)Zn binding on root of HE being available for loading into the xylem, as a result, more (65)Zn was translocated to the shoot.
Adsorption ; Biodegradation, Environmental ; Cells, Cultured ; Kinetics ; Metabolic Clearance Rate ; Plant Roots ; cytology ; metabolism ; Sedum ; cytology ; metabolism ; Zinc ; pharmacokinetics

Objective To build a multi‐level hierarchical structure model of the influencing factors for hierarchical medical system ,to identify the role relationship between all the factors and transmission pathways ,and to recommend on developing China′s hierarchical medical system . Methods Thirty influencing factors were identified in a screening based on literature review for the hierarchical medical system .On such basis ,16 influencing factors were identified by three health policy experts ,which affect operations of the current system .Interpretative structural modeling was called into play in the end to analyze the hierarchy relationship between various influencing factors and the conduction loops .Results There exist among the 16 factors a 3‐level hierarchical structureand two conduction loops .The factor directly limiting the hierarchical medical system is two‐way referral,and most internal core drivers arehuman resources development and governance mechanism.By means of self‐growth and external constraints ,they exert their influence on the operation of hierarchical medical system .Conclusions There are interactive hierarchical effects among the factors ,merging into three node elements of functional role,inter‐entity relationshipand patient participation.

Objective Radionuclide-labeled low molecular weight polypeptide is reeently advocated for the diagnosis and treatment of malignant tumor. The purpose of this study was to evaluate the anti-tumor effect of 131Ⅰ-Tyr-octreotide in nude mice bearing human non-small cell lung cancer (NSCLC). Methods 131Ⅰ-Tyr-octreotide was prepared by Ch-T method. The radiochemical purity was measured and biodistribution was evaluated. The nude mice models bearing human NSCLC were studied and divided into four groups: group A injected 131Ⅰ-Tyr-octreotide through tail vein, group B injected normal saline, group C injected 131Ⅰ-Tyroctreotide through stroma and group D injected 131Ⅰ through stroma. The radioactivity ratio of tumor to normal tissue (T/NT) was calculated over region of interest (ROI). The tumor cell cycle and cell apoptosis were analyzed by flow cytometry (FCM), terminal deoxynucleotidyl transferase mediated dUTP-biotion nick end labeling (TUNEL) and histopathological analysis. Statistical analysis was performed with SPSS 11.0, and the comparison for difference between groups performed with one-way ANOVA analysis. Results The labeled radiochemical purity was (95.23±1.67)% and specific activity of 3.5×106Bq/ug. The biodistributiou showed high uptake in kidney, and low uptake in liver and spleen. The radioactive uptake in group C was higher than the other groups, and the retention time was longer. The T/NT was 52.74±0.13 after 24 h, which was much higher than that the other groups (group D: 8.90±0.23, group A: 6.42±0.02, q=628.81 and 664.33, all P<0.05). The resuits of tmnor cell cycle determined by FCM showed that the G1 phase was blocked mast remarkably in group C than the other groups [group C: (83.17±6.86)%, group A: (57.02±18.81)%, group D: (49.29±7.80)%, group B: (45.88±5.13)%, q=5.29, 6.86, 7.55, 1.56, 2.26, 0.69, all P<0.05]. Apeptotic cells were observed by TUNEL, and apoptotic body was detected by immuno-histochemical examination. Conclusions 131Ⅰ-Tyr-octreotide was easily labeled by Ch-T. 131Ⅰ-Tyr-octreotide could induce tumor cell apoptosis and inhibit the tumor cell of NSCLC. It might be a potential target-directed agent in NSCLC.

OBJECTIVETo investigate the changes in whole brain gray matter volume in patients with trigeminal neuralgia using voxel-based morphometry (VBM).

METHODSTwenty-eight patients with trigeminal neuralgia and 28 healthy controls underwent magnetic resonance imaging with a Philips 1.5T MRI scanner. VBM was used to compare the structural differences in the whole brain gray matter between the two groups based on the DARTEL after data preprocessing with SPM8 software package.

RESULTSCompared with the healthy controls, the patients with trigeminal neuralgia presented with decreased gray matter volume in several brain regions including the bilateral middle temporal gyrus, bilateral superior/middle frontal gyrus, left pre-/post-central gyrus, right fusiform and anterior cingulate gyrus.

CONCLUSIONSPatients with trigeminal neuralgia had abnormal gray matter volume in some brain regions associated with perception and processing of pain sensation. These changes may provide clues for further exploration of the neuropathogenic basis of trigeminal neuralgia.

Brain Mapping ; Case-Control Studies ; Frontal Lobe ; pathology ; Gray Matter ; pathology ; Humans ; Magnetic Resonance Imaging ; Temporal Lobe ; pathology ; Trigeminal Neuralgia ; pathology