Extraperitoneal versus transperitoneal cesarean section: a retrospective study

To evaluate extraperitoneal cesarean section as a routine elective surgery. In this retrospective study, 461 primiparas were divided into the extraperitoneal and transperitoneal cesarean section groups according to the operation type in a random, but non-blinded, manner. The outcome measures were intraoperative blood loss, operation duration, postoperative gas passage time, postoperative pain, postoperative complications, and neonatal indicators The operation duration of the extraperitoneal cesarean section group was significantly lower than that of the transperitoneal cesarean section group (P  While extraperitoneal cesarean section can be safely performed as a routine procedure in the surgical delivery of primiparas, it must be performed by well-trained surgeons. In view of its advantages, it is worth being promoted in senior general hospitals as a routine choice. Abbreviations: CS: Cesarean section; ECS: Extraperitoneal; TCS: Transperitoneal; VAS: Visual analogue scale.</p

Additional file 1: of Handgrip strength is positively related to blood pressure and hypertension risk: results from the National Health and nutrition examination survey

Table S1. Association between handgrip strength and SBP adjusted for age and BMI stratified by gender. Table S2. Association between handgrip strength and DBP stratified by gender. (DOCX 15 kb

XLSearch: a Probabilistic Database Search Algorithm for Identifying Cross-Linked Peptides

Chemical cross-linking combined with mass spectrometric analysis has become an important technique for probing protein three-dimensional structure and protein–protein interactions. A key step in this process is the accurate identification and validation of cross-linked peptides from tandem mass spectra. The identification of cross-linked peptides, however, presents challenges related to the expanded nature of the search space (all pairs of peptides in a sequence database) and the fact that some peptide-spectrum matches (PSMs) contain one correct and one incorrect peptide but often receive scores that are comparable to those in which both peptides are correctly identified. To address these problems and improve detection of cross-linked peptides, we propose a new database search algorithm, XLSearch, for identifying cross-linked peptides. Our approach is based on a data-driven scoring scheme that independently estimates the probability of correctly identifying each individual peptide in the cross-link given knowledge of the correct or incorrect identification of the other peptide. These conditional probabilities are subsequently used to estimate the joint posterior probability that both peptides are correctly identified. Using the data from two previous cross-link studies, we show the effectiveness of this scoring scheme, particularly in distinguishing between true identifications and those containing one incorrect peptide. We also provide evidence that XLSearch achieves more identifications than two alternative methods at the same false discovery rate (availability: https://github.com/COL-IU/XLSearch)

Deguelin Induces Both Apoptosis and Autophagy in Cultured Head and Neck Squamous Cell Carcinoma Cells

<div><p>Head and neck squamous cell carcinoma (HNSCC) represents more than 5% of all cancers diagnosed annually in United States and around the world. Despite advances in the management of patients with this disease, the survival has not been significantly improved, and the search for potential alternative therapies is encouraging. Here we demonstrate that deguelin administration causes a significant HNSCC cell death. Deguelin induces both cell apoptosis and autophagy by modulating multiple signaling pathways in cultured HNSCC cells. Deguelin inhibits Akt signaling, and down-regulates survivin and cyclin-dependent kinase 4 (Cdk4) expressions, by disrupting their association with heat shock protein-90 (Hsp-90). Deguelin induces ceramide production through de novo synthase pathway to promote HNSCC cell death. Importantly, increased ceramide level activates AMP-activated protein kinase (AMPK), which then directly phosphorylates Ulk1 and eventually leads to cell autophagy. We found that a low dose of deguelin sensitized HNSCC cells to 5-FU. Finally, using a nude mice Hep-2 xenograft model, we also showed a significant anti-tumor ability of deguelin in vivo. Together, we suggest that deguelin may represent a novel and effective chemo-agent against HNSCC.</p> </div

Deguelin down-regulates survivin and Cdk4.

<p>Hep-2 cells were exposed to deguelin at indicated concentration (10, 25, 50 and 100 µM) for 24 hours, expression levels of survivin, Cdk4 and tubulin were measured by western blot (A).The association between survivin, Cdk4 and Hsp 90 as well as the association between survivin, Cdk4 and ubiquitin after deguelin treatment (Deg, 100 µM, 12 hours) were detected by immunoprecipitation (IP) assay (B–C). Effect of MG-132 (1 µM, 12 hour pretreatment) on deguelin-induced Cdk4 and survivin degradation was shown in (D). Western blots was performed to test the expression of Akt, phos-Akt, survivin, Cdk4 and tubulin after deguelin treatment in A253 (E) and SCC-9 cells (F). The mean of at least three independent experiments performed in triplicate is shown. Statistical significance was analyzed by ANOVA.</p

Deguelin activates AMPK signaling.

<p>Hep-2 cells were exposed to 100 µM of deguelin for indicated time points, followed by western blot detecting phospho- and total- level of LKB1-AMPK-ACC using specific antibodies (A). Hep-2 cells were transfected with scramble or AMPKα1RNAi (100 nM each) for 48 hours. Western blot was utilized to test AMPKα1 expression after transfection. Successfully AMPK knockdown cells and their parental cells were treated with deguelin (100 µM) in the presence or absence of AICAR (1 mM), phospho- and total- level of AMPKα1 were analyzed in (B–C), MTT assay was then used to test cell viability after 48 hours (D). Hep-2 cells were pretreated with fumonisin B1 (20 µM), C6-ceramide (10 µg/ml) or NAC (500 µM) for 1 hr before deguelin exposure for indicated time points, phospho- and total level of AMPKα1 as well as tubulin were tested by western blots (E–F). The mean of at least three independent experiments performed in triplicate is shown. Statistical significance was analyzed by ANOVA.</p

Deguelin’s <i>in vivo</i> anti-tumor effects in a Hep-2 xenograft model.

<p>Female BALB/c nude mice, 8–10 weeks of age, weighing 22 to 34 g, were acclimatized for 1 week before being injected s.c. with 3.5×10⁶ Hep-2 cells that had been re-suspended in 100 µL of medium. After 5 days when established tumors around 0.3 cm³ in diameter were detected, mice were randomized and divide in two groups. Ten mice per group were orally treated with deguelin (4 mg/kg) on days 1, 3, and 5 of each week for 3 weeks. Control animals received equal volume of saline solution. Tumor size was measured weekly using method mentioned above (A). Mice survival rate were also measured in (B). <i>p<0.05</i> vs. vehicle control group. Statistical significance was analyzed by ANOVA.</p

AMPK-dependent autophagy pathway contributes to deguelin-induced Hep-2 cell death.

<p>Hep-2 cells were pretreated with Z-VAD-fmk (50 µM) or 3-MA (2 mM) for 1 hr before deguelin exposure for 72 hours, cell viability was measured by MTT assay (A). Hep-2 cells were exposed to 100 µM of deguelin for indicated time points, followed by western blot detecting LC3B, tubulin, phospho- and total- level of Ulk1 (B). The association between AMPKα1 and Ulk1 after deguelin exposure (100 µM, 12 h) was determined by IP assay (C–D). Hep-2 cells were transfected with scramble or AMPKα1 siRNA (100 nM) for 48 hours. Western blot was then utilized to test AMPKα expression in transfected cells. Successfully AMPK knocking-down cells and their parental cells were treated with deguelin for indicated time points. AMPK, LC3B, tubulin, phospho- and total- level of Ulk1 were tested by western blots (E), LC3B and phospho-Ulk1 levels were quantified (F). Hep-2 cells were pretreated with 3-MA (2 mM) for 1 hr before deguelin (100 µM) exposure for 72 hours, cell apoptosis was measured by enzyme-linked immunosorbent cell apoptosis assay (G) and hoechst nuclear staining (H). (I) The proposed signaling pathway in this study: deguelin induces both cell apoptosis and autophagy by modulating multiple signaling pathways in cultured HNSCC cells: deguelin inhibits Akt signaling, and down-regulates survivin and cyclin-dependent kinase 4 (Cdk4) expressions, by disrupting their association with Hsp-90. Deguelin induces ceramide production through <i>de novo</i> synthase pathway to promote HNSCC cell death. Ceramide activates AMPK, which directly phosphorylates Ulk1 to promote an early cell autophagy. Cell autophagy is pro-apoptotic in our system. The mean of at least three independent experiments performed in triplicate is shown. Statistical significance was analyzed by ANOVA.</p

Deguelin induces cellular ceramide synthesis.

<p>Hep-2 cells were exposed to 100 µM of deguelin with or without fumonisin B1 (20 µM) for indicated time points, intracellular ceramide level was analyzed using methods mentioned above (A). Hep-2 cells were treated with fumonisin B1 (20 µM) or C6-ceramide (10 µg/ml), followed by deguelin (100 µM) exposure, MTT assay was used to test cell viability after 72 hours (B), Hoechst staining (C) and enzyme-linked immunosorbent cell apoptosis assay (D) were utilized to test Hep-2 cell apoptosis after 48 hours. The mean of at least three independent experiments performed in triplicate is shown. Statistical significance was analyzed by ANOVA.</p

Cytotoxic effects of deguelin in cultured HNSCC cells.

<p>Hep-2 cells were exposed to deguelin at a various concentration (10, 25, 50, 100 and 200 µM) for 72 hours (A) or exposed to 100 µM of deguelin for different time points (12, 24, 48 and 72 hours) (B), cell viability was then measured by MTT assay. (C) Three different cell lines A253, SCC-9 and PANC-1 were exposed to 100 µM of deguelin (Deg) for 72 hours; cell viability was measured by MTT assay. Hep-2 cells were treated with indicated concentration of deguelin, hoechst nuclear staining and enzyme-linked immunosorbent cell apoptosis assay were utilized to analyze cell apoptosis (D–E), caspase-3 activity was also measured (F), apoptosis related proteins including cleaved caspase-3, cleaved caspase-9, Bcl-Xl were detected by Western blots (G), Bcl-Xl expression level was quantified using image J software (H). The mean of at least three independent experiments performed in triplicate is shown. Statistical significance was analyzed by ANOVA. *<i>p</i><0.01 vs Control.</p