Comparison of long non-coding RNAs in adipose and muscle tissues between seven indigenous Chinese and the Yorkshire pig breeds.

lncRNAs play crucial roles in fat metabolism in animals. Previously, we have compared the mRNA transcriptome profiles between seven fat-type Chinese pig breeds and one lean-type Western breed (Yorkshire, YY). The associations between differentially expressed (DE) genes and phenotypical traits were investigated. In the present study, to further explore the underlying regulatory mechanisms, lncRNAs were sequenced and compared between YY and Chinese indigenous breeds. The results showed 9114 and 7538 DE lncRNAs between at least one Chinese breed and the YY breed in the adipose and muscle tissue respectively. KEGG enrichment analysis revealed that the target genes of these DE lncRNAs mainly influenced the glucolipid metabolism, which is an important process affecting meat quality. Correlation analyses between the DE lncRNA and DE mRNA genes related to meat quality and growth traits were performed. The results showed that LTCONS_00073280 was associated with intramuscular fat content. Four lncRNAs (LTCONS_00101781, LTCONS_00037879, LTCONS_00088260 and LTCONS-00128343) might mediate backfat thickness. Overall, this study provides candidate lncRNAs that potentially affect meat quality, which might be useful for molecular breeding of pig breeds in future.

[Clinical prognostic factors of adult supratentorial glioblastoma].

Objective: To analyze the treatment effect of patients with glioblastoma (GBM) and explore prognostic factors. Methods: The clinical data of 635 patients diagnosed as GBM at Neurosurgical Oncology Department Ⅳ of Beijing Tiantan Hospital, Capital Medical University from January 2007 to March 2018 were retrospectively reviewed. There were 386 males and 249 females with an age of (48.7±11.8) years (range: 18-75 years). Patients were divided into three groups according to the time of admission: 2007-2010 group(n=174), 2011-2014 group (n=237) and 2015-2018 group (n=224). Kaplan-Meier plot was used to analyze the effects of different treatment periods, treatment schemes and clinical factors on the survival of patients with GBM. Cox proportion hazard regression analysis was used to identify independent prognostic factors. Results: The median progression-free survival (PFS) and overall survival (OS) of patients in 2007-2010 group, 2011-2014 group, 2015-2018 group was 9.0 months (95% CI: 7.5-10.5), 10.0 months (95% CI: 8.8-11.2), 12.0 months (95% CI: 10.7-13.3) and 17.0 months (95% CI: 13.2-20.8), 20.0 months (95% CI: 16.9-23.1), 23.0 months(95% CI: 17.5-28.5), respectively. The PFS and OS of patients improved significantly over the years (χ(2)=9.693, P=0.008 and χ(2)=8.616, P=0.013). Multivariate survival analysis showed that age, extent of resection, radiotherapy and tumor distant dissemination were independent prognostic factors (all P<0.05). Conclusions: With the continuous development of clinical treatment regimen, the therapeutic effect of Chinese GBM patients has improved remarkably. Age, extent of resection, radiotherapy and tumor distant dissemination are independent prognostic factors associated with survival time.

Establishment of a novel triple-transgenic mouse: conditionally and liver-specifically expressing hepatitis C virus NS3/4A protease.

It is well known that NS3/4A protein plays crucial roles in the hepatitis C virus (HCV) replication. NS3/4A protein also results to virus-mediated immune evasion and persistence of infection through the interaction with host proteins. However, the lack of a suitable animal model hampers studies of HCV NS3/4A protein interaction with host proteins, which impacts immunopathology due to infection. Here, transgenic vector containing transcriptional regulation and Fluc reporter gene was constructed to conditionally express NS3/4A protein under the dual control of Tet-On regulatory system and Cre/LoxP gene-knockout system. NS3/4A transgenic founder mice were continuously crossed with Lap transgenic mice expressing reverse tetracycline-controlled transcriptional activator (rtTA), the NS3/4A/Lap double transgenic mouse lines with liver-specifically and conditionally expressing reporter (luciferase Fluc) under control of Tet-On system were established. The NS3/4A/Lap double transgenic mouse are mated with Lap/LC-1 double transgenic mouse with liver-specifically and conditionally expressing Cre recombinase under control of Tet-On system, NS3/4A/Lap/LC-1 triple transgenic mouse were generated. In vivo bioluminescent imaging, western blotting and immunohistochemical staining (IHS) was used to confirm that NS3/4A protein was strictly expressed in the liver of Doxycycline-induced triple transgenic mice. The results show that we established a triple-transgenic mouse model conditionally expressing the HCV NS3/4A protein under strict control of the Tet-On regulatory system and Cre/loxP system. This novel transgenic mouse model expressing NS3/4A in a temporally and spatially-specific manner will be useful for studying interactions between HCV NS3/4A protein and the host, also for evaluating NS3/4A protease inhibitors.

Stable color-tunable Ca3Y(GaO)3(BO3)4:Bi3+/Tb3+/Eu3+ phosphors for application in n-UV-pumped wLEDs.

For further development of light sources, white light-emitting diodes (wLEDs) have attracted widespread attention as promising next-generation light sources fabricated via the combination of phosphors and LED chips. However, latent defects, such as chemical/thermal instability, low color rendering index (CRI) and high correlated color temperature (CCT), of current mainstream wLEDs seriously hinder their further large-scale implementation. Herein, in order to overcome these limitations, single-phase color-tunable gaudefroyite (Ca3Y(GaO)3(BO3)4 (CYGB)) tridoped with Bi3+/Tb3+/Eu3+ ions was synthesized for the first time and detailed characterisation was performed via high-temperature solid-state reaction and structural/spectral analyses, respectively. Radius difference percentage calculations and Rietveld refinements indicate that dopants occupy both Y3+ and Ca2+ sites but preferably the Y3+ site over the Ca2+ site due to the same valence state. Through subtly regulating the (co)doping contents and skillfully utilizing the energy transfer (ET) strategy from the allowed transition of blue light-emitting Bi to the forbidden transition of green/red light-emitting Tb/Eu, the color hue (including white light) of highly efficient PL can be easily tuned according to the need. Meanwhile, composition/content-optimized white light-emitting CYGB:2%Bi/10%Tb/12%Eu also shows splendid chemical/thermal stability. Finally, as a proof-of-concept experiment, the CYGB:2%Bi/10%Tb/12%Eu phosphor-converted wLED (pc-wLED) was fabricated and encapsulated via the up-to-date remote 'capping' method, which imparted attractive performances. Altogether, the stable CYGB:Bi/Tb/Eu phosphor is a promising candidate for application in lighting/display fields.

Suitable internal control microRNA genes for measuring miRNA abundance in pig milk during different lactation periods.

Determination of an optimal set/number of internal control microRNA (miRNA) genes is a critical, but often undervalued, detail of quantitative gene expression analysis. No validated internal genes for miRNA quantitative PCR (q-PCR) in pig milk were available. We compared the expression stability of six porcine internal control miRNA genes in pig milk from different lactation periods (1 h, 3 days, 7 days, 14 days, 21 days, and 28 days postpartum), using an EvaGreen q-PCR approach. We found that using the three most stable internal control genes to calculate the normalization factor is sufficient for producing reliable q-PCR expression data. We also found that miRNAs are superior to ribosomal RNA (rRNA) and snRNA, which are commonly used as internal controls for normalizing miRNA q-PCR data. In terms of economic and experimental feasibility, we recommend the use of the three most stable internal control miRNA genes (miR-17, -107 and -103) for calculating the normalization factors for pig milk samples from different lactation periods. These results can be applied to future studies aimed at measuring miRNA abundance in porcine milk.

Properties of hepatitis C virus minigenome containing mutated 5'UTR region and luciferase transgene.

UNLABELLED: Sequences at the 3'UTR of Hepatitis C virus (HCV) negative-strand (-)RNA play an important role in the initiation of positive-strand (+)RNA synthesis. However, the underlying mechanism in cellular context is still unclear. In this report, we designed several cDNA-based HCV-like minigenomes containing different mutations at the 5'UTR of (+)RNA. These (+)RNAs transcribed from the minigenomes in vitro were transfected into HCV replicon cells for producing (-)RNAs with deletions of different stem loops (SL) at the 3'-end. The results showed that expression of the antisense transgene from minigenome increased, when the minigenome containing deletion of SL-C1+D1+E1 at the 3'-end of (-)RNA was transfected into the HCV replicon cells compared to that of the full minigenome. The expression of the transgene from minigenome decreased using other mutant minigenomes containing deletions SL-A1, SL-A1+B1, and SL-A1+B1+C1 at the 3'-end of (-)RNA. Finally, the transgene from SL-C1+D1+E1 of (-)RNA using CMV promoter-driven minigenome was expressed at higher level than full minigenome in HCV replicon cell lines. These results indicated that the region of (-)RNA interacting with HCV replicase may locate in the SL-C1+D1+E1 region of (-)RNA. KEYWORDS: Hepatitis C virus; minigenome; RNA dependent RNA polymerase; replication.

Scavenger receptor class B type I mediates cell entry of hepatitis C virus.

This study assessed the functional role of human scavenger receptor class B type I (SR-BI) as a putative hepatitis C virus (HCV) receptor using Chinese hamster ovary (CHO) cells transfected with human SR-BI (CHO-huSR-BI). The expression of SR-BI by primary Tupaia hepatocytes (PTHs), human hepatocarcinoma cell line (HepG2) cells, untransfected CHO cells and CHO-huSR-BI cells was analysed by Western blotting. Receptor competition assays showed that anti-SR-BI antibodies that block the binding of soluble envelope glycoprotein E2 could prevent HCV infection. Pre-incubation of CHO-huSR-BI and HepG2 cells with anti-SR-BI antibodies resulted in marked inhibition of E2 binding. After incubation with HCV RNA-positive serum from a patient with chronic HCV infection, however, HCV infection could not be detected in CHO-huSR-BI cells, but was detected in PTHs. These results demonstrate that, whilst SR-BI represents an important cell surface molecule for HCV infection, the presence of SR-BI alone is insufficient for HCV entry.

Development of a cell-based assay for monitoring hepatitis C virus ns3/4a protease activity.

Hepatitis C virus (HCV) NS3/4A (non-structural 3 and 4 B) protease plays a key role in the processing of polyprotein precursor and it becomes an attractive target for antiviral drug discovery. We developed a cell-based assay for monitoring of the NS3/4A protease activity in mammalian cells that is an important step in screening of specific drugs against the protease. The recombinant caspase 3 (rCasp3) was used as the specific substrate for NS3/4A protease. The endogenous cleavage sites in the procaspase 3 molecule were substituted by decapeptides specific for NS3/4A protease. The activation of rCasp3 depended on its specific cleavage by NS3/4A protease and resulted in an apoptosis of stable cells expressing the protease. The difference in cell viability between the cells expressing NS3/4A protease transfected with rCasp3 and the counterparts pretreated with NS3/4A protease inhibitors could be estimated by a spectrophotometry based on 3-(4,5-dimethylthioazol- 2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) staining of cells in microplates. Thus, we developed a simple and cost-effective colorimetric assay for evaluating NS3/4A protease activity enabling the screening of candidate NS3/4A protease inhibitors.

Mesoporous nanoparticles Gd2O3@mSiO2/ZnGa2O4:Cr3+,Bi3+ as multifunctional probes for bioimaging.

Red/near infrared (NIR) persistent luminescent nanoparticles (PLNPs) hold great potential as a new generation of probes for the detection and imaging of biomolecules. Based upon the consideration that a single nanoprobe could serve multiple purposes, the development of a multimodal nanoprobe that combined the properties of rechargeable persistent emitting luminescence, magnetic resonance imaging (MRI) and drug delivery has attracted our attention as a promising prospect in the field of nanotechnology directed toward biomedical applications. Herein, Gd2O3@mSiO2/ZnGa2O4:Cr3+,Bi3+ (ZGOCB) mesoporous nanoparticles that exhibit enhancement of red (∼695 nm) persistent luminescence (∼18 d) properties were synthesized by using mesoporous silica nanospheres both as morphology-controlling templates and vessels. Being composed of hybrid shell/core architecture and through surface functionalization, Gd2O3@mSiO2/ZGOCB mesoporous nanoparticles possess the capacity for in vivo and in situ real-time monitoring, targeting tumors and drug delivery. Simultaneously, Gd2O3@mSiO2/ZGOCB exhibits a prominent longitudinal relaxivity, indicating that these nanoparticles could also be used as magnetic resonance imaging agents. We believe that this rechargeable red persistent luminescence and MRI-based core/shell structure of the multimodal nanoprobe offers a promising nano-platform for both diagnostics and therapeutics of reactive species in living cells or in vivo.

[Radiotherapy of palpebral carcinoma--a follow-up study].

After radiotherapy, 52 patients of palpebral carcinoma were followed up, including squamous cell carcinoma 24 cases, basal cell carcinoma 19 cases, undifferentiated carcinoma 7 cases, and Meibomian gland carcinoma 2 cases. The tumors of 46 cases (88.5%) resolved in 3-6 months after treatment, and the 3-year survival rate was 92.3% (48 cases), the 5-year survival rate was 82.7% (43 cases), and the 10 to 16-year survival rate was 47.7% (21 cases). The curative effect of radiotherapy on palpebral carcinoma is noteworthy.

A novel hepatitis C virus vaccine approach using recombinant Bacillus Calmette-Guerin expressing multi-epitope antigen.

Hepatitis C virus (HCV) is a major cause of liver disease worldwide. HCV infection is associated with high morbidity and has become a major problem in public health. Until now, there has been no effective prophylactic or therapeutic vaccine. BCG, a live vaccine typically used for tuberculosis prevention, has been increasingly utilized as a vector for the expression of recombinant proteins that will induce specific humoral and cellular immune responses. In this study, recombinant BCG (rBCG) was engineered to express a HCV multi-epitope antigen CtEm, and HLA-A2.1 transgenic mice were immunized with rBCG-CtEm. High levels of specific anti-HCV antibodies targeted to mimotopes of HVR1 were detected in the serum. HCV-specific lymphocyte proliferation assay, cytokine determination and cytotoxicity assay indicated that HCV epitope-specific cellular immune responses were elicited in vitro. The rBCG-CtEm immunization conferred protection against infection with the recombinant vaccinia virus (rVV-HCV-CNS) in vivo. These results suggest that rBCG expressing multi-epitope antigen may serve as an effective vaccine against HCV infection.