Construction of multiple shRNAs expression vector that inhibits FUT1 gene expression and production of the transgenic SCNT embryos in vitro.

Enterotoxigenic Escherichia coli F18 is a major pathogen that causes postweaning diarrhoea and edema disease in piglets. The alpha(1,2)-fucosyltransferase (FUT1) gene has been identified as an ideal candidate gene for controlling the expression of the receptor for ECF18 bacteria. Therefore, the use of RNA interference (RNAi) to study the function of the FUT1 gene and to produce FUT1 knockdown transgenic pig would be highly beneficial. We developed an effective strategy for the expression of multiple small hairpin RNA simultaneously using multiple RNA polymerase III (hU6, hH1, mU6 and h7SK) promoters in a single vector to knockdown the FUT1 gene. Stable FUT1 knockdown transgenic fibroblast lines were generated by transfecting porcine fetal fibroblasts with the constructed vectors. Real-time RT-PCR indicated that the mRNA level of FUT1 in the transgenic fibroblast lines was significantly lower than that in the control, as much as 29 %. Finally, we successfully obtained transgenic SCNT porcine embryos. Overall, the results demonstrated that this vector-based RNAi expression system is an efficient approach to knockdown FUT1 gene expression in porcine fetal fibroblast cells, which could thereby provide donor cells for somatic cell nuclear cloning and the potential production of a marker-free transgenic pig resistant to F18 related diseases. Furthermore, it also provides strong evidence that this approach could be useful both in the production of transgenic livestock resistant to disease, and in the development of effective strategies for the suppression of gene expression in clinical gene therapy.

Effect of a hybrid team-based advanced cardiopulmonary life support simulation program for clinical nurses.

BACKGROUND: During in-hospital cardiac arrest events, clinical nurses are often the first responders; therefore, nurses require sufficient advanced cardiac life support (ACLS) competency. This study aimed to verify the effects of a hybrid team-based ACLS simulation (HTAS) program (developed in this study) on nurses' ACLS performance, specifically ACLS knowledge, cardiopulmonary resuscitation (CPR) self-efficacy, and CPR-related stress. METHODS: The developed HTAS comprised four lecture videos, one team-based skills training video, and a team-based ACLS simulation. A quasi-experimental pretest-posttest design with a comparison group (CG) was used to evaluate the effectiveness of the HTAS. Of the 226 general ward nurses with more than 6 months of clinical experience, 117 were allocated to the intervention group (IG), which attended the HTAS, and 109 to the CG, which attended only basic ACLS training. RESULTS: The IG's ACLS performance significantly improved (t = 50.8, p < 0.001) after the training. Relative to the respective pretest conditions, posttest ACLS knowledge (t = 6.92, p < 0.001) and CPR self-efficacy (t = 6.97, p < 0.001) of the IG also significantly increased. However, when the mean difference values were compared, there was no significant difference between the two groups with respect to ACLS knowledge (t = 1.52, p = 0.130), CPR self-efficacy (t = -0.42, p = 0.673), and CPR stress (t = -0.88, p = 0.378). CONCLUSION: The HTAS for ward nurses was effective at enhancing the nurses' ACLS performance. It is necessary to develop effective training methods for team-based ACLS and verify the sustained effects of such training.

TcpC inhibits neutrophil extracellular trap formation by enhancing ubiquitination mediated degradation of peptidylarginine deiminase 4.

TcpC is a multifunctional virulence factor of uropathogenic E. coli (UPEC). Neutrophil extracellular trap formation (NETosis) is a crucial anti-infection mechanism of neutrophils. Here we show the influence of TcpC on NETosis and related mechanisms. We show NETosis in the context of a pyelonephritis mouse model induced by TcpC-secreting wild-type E. coli CFT073 (CFT073wt) and LPS-induced in vitro NETosis with CFT073wt or recombinant TcpC (rTcpC)-treated neutrophils are inhibited. rTcpC enters neutrophils through caveolin-mediated endocytosis and inhibits LPS-induced production of ROS, proinflammatory cytokines and protein but not mRNA levels of peptidylarginine deiminase 4 (PAD4). rTcpC treatment enhances PAD4 ubiquitination and accumulation in proteasomes. Moreover, in vitro ubiquitination kit analyses show that TcpC is a PAD4-targetd E3 ubiquitin-ligase. These data suggest that TcpC inhibits NETosis primarily by serving as an E3 ligase that promotes degradation of PAD4. Our findings provide a novel mechanism underlying TcpC-mediated innate immune evasion.

Robust and nanostructured chitosan-silica hybrids for bone repair application.

In this study, chitosan-silica hybrids (CSHs) with superior mechanical strength and homogeneous dispersion of nano-sized silica particles were synthesized via a facile sol-gel method aiming for bone regeneration. The effects of varied acidic conditions of sol-gel reaction and inorganic/organic ratios on the performance of the hybrid were investigated. CSHs synthesized under weak acidic conditions (acetic acid, pH 4.0) showed a homogeneous nanostructure and robust strength (maximum compressive strength: 42.6 ± 3.3 MPa and 271 ± 31 MPa in wet and dry forms, respectively). However, those developed under the strong acidic condition (HCl, pH 4.0) and the strong acid condition plus lower pH (HCl, pH 2.8) tended to aggregate and exhibited inferior mechanical properties (compressive strength: 6.3 ± 0.3 MPa in wet form at pH 2.8). Under the latter conditions, the interactions between silica and chitosan were weak. Moreover, the mechanical properties of the CSHs could be tuned in a wide range by conveniently varying the inorganic/organic composition ratio between 50% and 70%. In vitro cytocompatibility study indicated that CSHs were non-cytotoxic. These results suggested that the weak acidic sol-gel process were essential for fabricating chitosan-silica hybrids with high mechanical strength, which had potential to be applied as a bone substitute.

Clinical effect of laparoscopic splenectomy and pericardial devascularization in portal hypertension patients with Child-Pugh A/B liver function / 临床肝胆病杂志

ObjectiveTo investigate the clinical effect of laparoscopic splenectomy and pericardial devascularization (LSPD) in patients with portal hypertension and the long-term effect of LSPD. MethodsA total of 40 portal hypertension patients with Child-Pugh A/B liver function who received LSPD in The First Hospital of Jilin University from August to December 2017 were enrolled as surgical group, and 44 portal hypertension patients with Child-Pugh A/B liver function who received conservative treatment during the same period of time was enrolled as internal medicine group. The patients were followed up to June 30, 2019, and liver function parameters, upper gastrointestinal bleeding, and portal vein thrombosis were recorded for all patients at each time point. The t-test was used for comparison of normally distributed continuous data between two groups; an analysis of variance was used for comparison between multiple groups, and the Bonferroni test was used for further comparison between two groups. The Kruskal-Wallis H test was used for comparison of continuous data with skewed distribution; between multiple groups, and the Mann-Whitney U test was used for further comparison between two groups. The chi-square test was used for comparison of categorical data between groups. ResultsAt 6, 12, and 24 months after discharge, compared with the internal medicine group, the surgical group had a significantly higher level of cholinesterase (t=3.527, 3.849, and 5.555, all P＜0.05) and a significantly lower Child-Pugh score (t=2.498, 2.138, and 2.081, all P＜0.05). Compared with the internal medicine group at 12 and 24 months after discharge, the surgical group had a significantly higher level of albumin (t=3.120 and 2.587, both P＜0.05) and a significantly lower incidence rate of upper gastrointestinal bleeding (χ2=4.947 and 5.155, both P＜0.05). At 24 months after discharge, the surgical group had a significantly lower number of patients who had a significant increase in alpha-fetoprotein level than the internal medicine group (χ2=4.648, P=0.031). At 12 months after discharge, the surgical group had a significantly higher incidence rate of portal vein thrombosis than the internal medicine group (χ2=4.395, P=0.036). The surgical group had significant improvements in albumin (F=2.959, P=0.013), cholinesterase (F=11.022, P＜0001), prothrombin time (H=94.100, P＜0.001), and Child-Pugh score (F=3.742, P=0.003) from admission to 12 and 24 months after surgery. ConclusionIn portal hypertension patients with Child-Pugh A/B liver function, LSPD can improve liver function and reduce the incidence rate of upper gastrointestinal bleeding, and the high incidence rate of portal vein thrombosis can be effectively reduced by oral aspirin and rivaroxaban.