The Improved Biocontrol Agent, F1-35, Protects Watermelon against Fusarium Wilt by Triggering Jasmonic Acid and Ethylene Pathways.

Watermelon Fusarium wilt, caused by Fusarium oxysporum f. sp. niveum (FON), is one of the most important diseases, and has become a major limiting factor to watermelon production worldwide. Previous research has found that the improved biocontrol agent, F1-35, had a high control efficiency to watermelon Fusarium wilt. In this study, the control efficiency of F1-35 to watermelon Fusarium wilt was firstly tested, and the control efficiency was 61.7%. Then, we investigated the mode of action of F1-35 in controlling watermelon Fusarium wilt. Using a pairing assay, we found that F1-35 did not inhibit the normal growth of FON. To know more about the interaction between F1-35 and watermelon root, the protein expressions of roots after 12, 24, and 48 h post-inoculation were examined. A total of 1109 differentially expressed proteins were obtained. KEGG analysis found that the most differentially expressed proteins occurred in alpha-linolenic acid metabolism, cysteine and methionine metabolism, plant-pathogen interaction, and the MAPK signaling pathway to the plant. A further analysis of differentially expressed proteins showed that F1-35 triggered the jasmonic acid and ethylene pathways in watermelon. To validate our results, the qRT-PCR was used to analyze the gene expression levels of PAL, LOX1, and CTR1. The gene expression results showed that those genes, which were positive correlated with the JA pathway, were up-expressed, including PAL and LOX1, and the negative associated gene, CTR1, was down-expressed. In conclusion, the improved biocontrol agent, F1-35, improves the resistance of watermelons to FON by triggering the JA and ET pathways.

[Case-control study on a new Kirschner wire tension band with hole in the treatment of adult olecranon fractures in adults].

OBJECTIVE: To compare the efficacy of Kirschner wire tension band with hole and common Kirschner wire tension band in the treatment of olecranon fractures in adults, so as to guide the selection of clinical surgical procedures. METHODS: From July 2011 to October 2015, a total of 49 patients with olecranon fractures underwent open reduction and fixation with Kirschner wire tension band, which were retrospectively analyzed. Of the 49 patients, 21 patients(group A) were treated with Kirschner wire tension band with hole, including 12 males and 9 females, with an average age of(37.6 ±8.2) years old;28 patients(group B) were treated with common Kirschner wire, including 18 males and 10 females, with an average age of(38.9±7.3) years old. There were no statistically significant differences between the two groups in general data. The differences of operative duration, frequency of radiation, fracture healing time, complications and postoperative elbow function scores between the two groups were observed and analyzed by parallel statistical analysis. RESULTS: In group A, the operative duration was(60.4±10.7) min, the average number of radiation times was (12.5±2.9); in group B, the operative duration was (62.3±11.8) min, and the average radiation times was(13.7±3.8); there was no significant difference in operative time and radiation times between the two groups (P >0.05). In group A, the fracture healing time was (13.2±2.6) weeks without K-wire migration, skin irritation and other complications; and fracture healing time in group B was(14.6±1.8) weeks with complications(K-wire migration in 6 cases, skin irritation in 7 cases, internal fixation failure in 2 cases);the fracture healing time of group A was shorter than that in group B. Evaluation of elbow joint function in two groups of patients after operation showed that in group A, pain score was 41.0±3.5, movement function score was 18.0±2.1, stability score was 9.0±0.8, daily activities score was 18.0±4.3, the total average score was 87.0±7.8; and in group B, the pain score was 39.0±5.6, movement function score was 17.0±3.2, the stability score was 8.0±2.4, daily activities score was 16.0±5.2, the total average score was 83.0±10.7. There were no statistically significant in the scores between the two groups. CONCLUSIONS: Compared with ordinary Kirschner wire, the treatment of olecranon fracture with Kirschner tension band with hole can shorten the time of fracture healing, significantly reduce the occurrence of complications, and do not affect the recovery of postoperative function, which is suitable for clinical use.

Early prediction of organ failure under the revised Atlanta classification.

BACKGROUND/AIMS: This study aimed to compare the ability of conventional laboratory markers and scoring systems to early predict organ failure (OF) and to differentiate between transient and persistent OF in patients with acute pancreatitis (AP) using the revised Atlanta classification. MATERIALS AND METHODS: We retrospectively analyzed the medical records of 214 patients with AP between January 2014 and July 2015. The predictive values of laboratory markers were analyzed. The predictive accuracy of individual markers, extrapancreatic inflammation on computed tomography (EPIC), acute physiology and chronic health evaluation II (APACHE II), and bedside index for severity in acute pancreatitis (BISAP) scores were measured using the area under the receiver operating characteristic curve (AUROC). RESULTS: OF was diagnosed in 32 (15%) patients and persistent OF in 14 (6.5%). There were statistically significant differences between patients with and without OF with respect to white blood cell count, creatinine, blood urea nitrogen, lactate dehydrogenase, C-reactive protein, calcium (Ca), arterial partial pressure of oxygen (PaO2), base excess (BE), APACHE II, BISAP scores, and EPIC scores. Logistic regression analysis identified Ca, PaO2, and BE as independent predictors of OF. Using AUROC, the EPIC score had the highest accuracy for the early prediction of OF, which was 0.82. No significant differences were detected between patients with transient and persistent OF. CONCLUSION: Several laboratory markers and score systems were useful for the early prediction of OF in patients with AP, of which Ca, PaO2, and BE had highest predicting value, and EPIC score had the highest accuracy. We could not predict the duration of OF using laboratory markers.

Inhibitory effects of paeoniflorin on lysophosphatidylcholine-induced inflammatory factor production in human umbilical vein endothelial cells.

Lysophosphatidylcholine (LPC) plays an important role in atherosclerosis through initiation of endothelial inflammation response. Paeoniflorin (PEF), isolated from the dry root of Paeonia, has been reported to exert an anti-inflammatory effect, but the exact mechanism is not fully understood. The aim of this study was to investigate the inhibitory effects of PEF on LPC-induced inflammatory factor production and the underlying mechanisms. In human umbilical vein endothelial cells (HUVECs), different concentrations (1, 10 or 100 µmol/l) of PEF were added 2 h prior to exposure to LPC (10 mg/l) for 24 h. The results showed that PEF significantly inhibited LPC-induced inflammatory factor production. In addition, PEF was also able to suppress the enhanced high mobility group box-1 (HMGB1) expression and release, upregulated expression of receptor for advanced glycation end product (RAGE), Toll-like receptor (TLR)-2 and TLR-4, and increased nuclear factor-κB (NF-κB) activity induced by LPC. Our results suggest that PEF suppresses LPC-induced inflammatory factor production through inhibition of the HMGB1-RAGE/TLR-2/TLR-4-NF-κB pathway.

Paeoniflorin protects myocardial cell from doxorubicin-induced apoptosis through inhibition of NADPH oxidase.

Increased intracellular reactive oxygen species (ROS) are involved in doxorubicin (DOX)-induced myocardial cell apoptosis, and paeoniflorin (PEF) has been shown to exert an antioxidant effect. The aim of the present study was to explore the protective effect of PEF on DOX-induced myocardial cell apoptosis and the underlying mechanisms. In cultured H9c2 cells, different concentrations (1, 10, or 100 µmol/L) of PEF was added for 2 h prior to exposure to DOX (5 µmol/L) for 24 h. Cell apoptosis was evaluated by hoechst 33342 staining, and caspase-3 expression and activity. The mRNA and protein expression of NADPH oxidase (NOX) 2 and NOX4 was determined by real-time polymerase chain reaction and Western blot, respectively. Intracellular ROS and NOX activity were measured by assay kit. The results showed that DOX significantly increased myocardial cell apoptosis, increased caspase-3 expression and activity concomitantly with enhanced ROS production, and increased NOX2, NOX4 mRNA and protein expression, and NOX activity. These effects were remarkably inhibited by pretreatment of PEF. Our results suggested that PEF has a protective effect against DOX-induced myocardial cell apoptosis through a mechanism involving a decrease in ROS production by inhibition of NOX2, NOX4 expression, and NOX activity.

Covalent conjugation of avidin with dye-doped silica nanopaticles and preparation of high density avidin nanoparticles as photostable bioprobes.

Progress in biomedical imaging depends on the development of bioprobes with a high sensitivity and stability. Fluorescent silica nanoparticles (NPs) covalent conjugation of avidin has been proposed for cancer cells imaging by fluorescence microscopy. Uniform silica NPs were prepared using water-in-oil (W/O) microemulsion methods and primary amine groups were introduced onto the surface of the NPs by condensation of tetraethyl orthosilicate (TEOS). Optically stable organic dyes, tris(2,2'-bipyridyl) dichlororuthenium(II) hexahydrate (Rubpy), were doped inside the silica NPs. The amine functions were transferred to carboxyl groups coupled with a linker elongation. Avidin was immobilized at the surface of the NPs by covalent binding to the carboxyl linkers. The binding capacity of the avidin-covered NPs for ligand biotin was quantified by titration with biotin(5-fluorescein) conjugate to 1.25 biotin binding sites/100 nm(2). We used biotinylated antibody and cell recognition by fluorescence microscopy imaging technique. The lung carcinoma cells were identified easily with high efficiency using these antibody-coated NPs. By comparison with fluorescein isothiocyanate (FITC), dye-doped silica NPs display dramatically increased stability of fluorescence as well as photostability, as compared to the common organic dye, when under continuous irradiation.

Silica nanoparticles based label-free aptamer hybridization for ATP detection using hoechst33258 as the signal reporter.

In this work, we have developed a simple and sensitive method for ATP detection using silica nanoparticles (NPs) as the platform and hoechst33258 as the signal reporter. The ATP-binding aptamers hybridize with the probe DNA (DNA(p)) immobilized NPs to form the aptamer/DNA(p) duplex on the NPs surface. The conformational change of the aptamer leads to the decrease of the aptamer/DNA(p) duplex on the NPs due to the ATP-binding aptamer switches its structure from the aptamer/DNA(p) duplex to the aptamer/target complex in the presence of ATP. ATP detection can be easily realized by separating the silica nanoparticles and adding the hoechst33258 of intercalating to aptamer/DNA(p) (dsDNA). Good selectivity between ATP and CTP, GTP or UTP has been demonstrated, which is due to the specific recognition between ATP aptamer and ATP. The K(d) was estimated to be ∼1mM from 0 to 4mM and a liner response was observed from 0 to 0.2mM with a detection limit of ∼20µM. Compared with other methods, the carboxyl-modified silica nanoparticles (∼60nm) prepared by the reverse microemulsion method can serve as a stable and sensitive sensor platform because of their smaller size and facile conjugation with amine-containing molecules. In addition, the high sensitivity and selectivity of hoechst33258 was employed for the ssDNA and dsDNA determination, which takes advantage of the label-free aptamer and lower cost.

A new cytotoxic flavonoid from the fruit of Sinopodophyllum hexandrum.

Constituents of the fruit of Sinopodophyllum hexandrum (Royle) Ying (Sinopodophylli Fructus) were investigated. A new flavonoid, 8,2'-diprenylquercetin 3-methyl ether along with 9 known compounds were isolated and identified. Among them, the new compound 8,2'-diprenylquercetin 3-methyl ether exhibited cytotoxic activities against MDA-231 and T47D breast cancer cell lines, quercetin, kaempferol and rutin were isolated from Sinopodophylli Fructus for the first time.

[Immune regulating activity of a novel organoselenium compound ethaselen-1 in C57/BL mice].

OBJECTIVE: To detect the Immune regulating activity of ethaselen-1 (Eb1), a novel organoselenium compound, in C57/BL mice transplanted with Lewis lung cancer(LLC). METHODS: The LLC transplanted C57/BL mice models were established, and the mice was randomly divided into four groups, including high dose Eb1 group (25.0 mg/kg),low dose Eb1 group (12.5 mg/kg), positive control group (levamisole, LMS, 2.0 mg/kg) and negative control group(solvent). Intraperitoneal injections (ip.) of the four pharmaceuticals were performed once a day through the abdominal wall separately, from the second to the eighth day after cancer was transplanted. On the eleventh day, six mice of each group were killed and relative weight of spleen, transforming activity of spleen lymphocytes, NK cell activity, LAK cell activity and percentage of CD4(+)CD8(+)T lymphocyte were detect. RESULTS: Compared with the control group, high dose Eb1 could obviously increase the relative weight of spleen (150.59% and 122.55%), transforming activity of spleen lymphocytes(162.25% and 561.98%), NK cell activity(78.60% and 219.42%) and percentage of CD4(-)/CD8(+) T lymphocyte(104.72% and 105.87%) in normal mice and LLC mice. Compared with the control group, high dose Eb1 could also increase LAK cell activity of LLC mice by 195.11%. CONCLUSION: The novel organoselenium compound Eb1 has immune regulating activity in vivo.

[Development of a novel in-vitro culture device simulating distraction osteogenesis and expression of osteoblast cytoskeleton].

OBJECTIVE: To establish a novel in-vitro culture device simulating distraction osteogenesis, and to understand the the changes of cytoskeleton of osteoblasts under different magnitudes of mechanical distraction. METHODS: A novel in-vitro culture device simulating distraction osteogenesis was developed with titanium and silicon membrane. The expressions of F-actin cytoskeleton subjected to different magnitudes of distraction were observed in situ under a confocal laser scanning microscope, and protein levels of actin and tubulin alpha were analyzed by electrophoresis of Western blot. RESULTS: The mode of culture device simulating distraction osteogenesis was similar to that of distraction osteogenesis used in clinic. After 24 h mechanical distraction with physiological magnitude, F-actin of osteoblasts was organized and paralleled to the direction of distraction forces. After 24 h mechanical distraction with hyperphysiological magnitude, however, F-actin of osteoblasts became reorganized and disrupted, and protein levels of actin and tubulin alpha decreased. CONCLUSIONS: The novel in vitro culture device simulating distraction osteogenesis is suitable for the study of distraction osteogenesis in vitro. Physiological mechanical distraction causes organization and rearrangement of actin cytoskeleton. However, hyperphysiological mechanical distraction results in reorganization and disruption of actin cytoskeleton.

[The antitumor activity of Shuang-Xi-Zuo-Wan-1 in C57/BL mice].

OBJECTIVE: To detect the antitumor activity of Shuang-Xi-Zuo-Wan-1(Eb), a novel organoselenium compound, in C57/BL mice transplanted with Lewis lung cancer(LLC). METHODS: The LLC transplanted C57/BL mice model was established,and the mice were randomly divided into four groups, including high dose Eb group (25.0 mg/kg), low dose Eb group (12.5 mg/kg), positive control group (DDP, 2.0 mg/kg) and negative control group (solvent). Each group had twelve mice. Intraperitoneal injections (ip.) of four pharmaceuticals were performed once a day through the abdominal wall separately, from the second to the eighth days after cancer was transplanted. On the eleventh day, six mice of each group were killed and the influences of Eb on growth speed, size, weight, invasion, inhibitory rate, proliferation index and apoptosis rate of LLC were observed and calculated. The remaining mice were fed till all of them died naturally and the average survival time of each group was calculated. RESULTS: Eb could inhibit the growth and infiltration of LLC (the cancer inhibitory rate of high does Eb was 80.31%) obviously and prolong the average survival time of these with mice cancer. After being given Eb, the nuclear of the cancer cell concentrated and the fission phase cells reduced. In addition,the number of apoptosis cancer cells increased. CONCLUSION: The novel organoselenium compound Eb has antitumor activity in vivo. It can inhibit the growth and infiltration of LLC in mice, and induce the apoptosis of cancer cells.

[Culture of osteoblasts by serial explant culture and comparison of their characteristics].

OBJECTIVE: To Investigate a simple, economical and efficient method of primary osteoblast culture and compare their characteristics. METHODS: Primary osteoblasts of 1 st-4 th series explant culture from calvarial bones of neonatal Sprague-Dawley rats, were collected and osteoblasts shape, mitosis, proliferation, ALP activity and immunohistological expression of osteocalcin and BMP-2 observed. RESULTS: No difference was found in cell shape, spread, confluence, ALP activity as well as immunohistological stain of osteocalcin and BMP-2 of the osteoblasts, which had been harvested from the 1 st-4 th series explant cultures of Sprague-Dawley rats. CONCLUSION: Series explant culture can harvest the same kind of osteoblasts as the explant culture, and more osteoblasts can be obtained at a single time. It saves money and time and is easy to manipulate.