Advances in MicroRNA-Mediated Reprogramming Technology

The use of somatic cells to generate induced-pluripotent stem cells (iPSCs), which have gene characteristic resembling those of human embryonic stem cells (hESCs), has opened up a new avenue to produce patient-specific stem cells for regenerative medicine. MicroRNAs (miRNAs) have gained much attention over the past few years due to their pivotal role in many biological activites, including metabolism, host immunity, and cancer. Soon after the discovery of embryonic-stem-cell- (ESC-) specific miRNAs, researchers began to investigate their functions in embryonic development and differentiation, as well as their potential roles in somatic cell reprogramming (SCR). Several approaches for ESC-specific miRNA-mediated reprogramming have been developed using cancer and somatic cells to generate ESC-like cells with similarity to iPSCs and/or hESCs. However, the use of virus-integration to introduce reprogramming factors limits future clinical applications. This paper discusses the possible underlying mechanism for miRNA-mediated somatic cell reprogramming and the approaches used by different groups to induce iPSCs with miRNAs

Intronic MicroRNA (miRNA)

Nearly 97% of the human genome is composed of noncoding DNA, which varies from one species to another. Changes in these sequences often manifest themselves in clinical and circumstantial malfunction. Numerous genes in these non-protein-coding regions encode microRNAs, which are responsible for RNA-mediated gene silencing through RNA interference (RNAi)-like pathways. MicroRNAs (miRNAs), small single-stranded regulatory RNAs capable of interfering with intracellular messenger RNAs (mRNAs) with complete or partial complementarity, are useful for the design of new therapies against cancer polymorphisms and viral mutations. Currently, many varieties of miRNA are widely reported in plants, animals, and even microbes. Intron-derived microRNA (Id-miRNA) is a new class of miRNA derived from the processing of gene introns. The intronic miRNA requires type-II RNA polymerases (Pol-II) and spliceosomal components for their biogenesis. Several kinds of Id-miRNA have been identified in C elegans, mouse, and human cells; however, neither function nor application has been reported. Here, we show for the first time that intron-derived miRNAs are able to induce RNA interference in not only human and mouse cells, but in also zebrafish, chicken embryos, and adult mice, demonstrating the evolutionary preservation of intron-mediated gene silencing via functional miRNA in cell and in vivo. These findings suggest an intracellular miRNA-mediated gene regulatory system, fine-tuning the degradation of protein-coding messenger RNAs

Polimorfizm rs10830963 w genie receptora melatoniny 1B a cukrzyca ciążowa w populacji chińskiej — metaanaliza badań

Introduction: Studies have been conducted to investigate the association between rs10830963 of MTNR1B and the risk of gestational diabetes mellitus (GDM), but with inconclusive results. We aimed to clarify these controversies, especially with regard to the association in the Chinese population. Material and methods: A systemic literature reference search inclusive to August 12, 2016 yielded 35 articles, from which 11 studies met the inclusion criteria for the final meta-analysis, including 3889 patients with GDM and 6708 controls. Results: We found statistically significant associations between rs10830963 and GDM using odds ratios (ORs) and 95% confidence intervals (CIs) [GG genotype vs. CC genotype: OR = 1.70, 95% CI: 1.38–2.10; G allele vs C allele: OR = 1.27, 95% CI: 1.20–1.36; GG+CG vs. CC (dominant model): OR = 1.31, 95% CI: 1.20–1.44; GG vs CG+CC (recessive model): OR = 1.41, 95% CI: 1.26–1.58]. In subgroup analyses stratified by ethnicity, we also observed rs10830963 to be associated with significantly increased risk of GDM in all genetic models in the Chinese population. Conclusions: Our meta-analysis indicated that the rs10830963 polymorphism might serve as a risk factor of GDM in the Chinese population.Wstęp: Wyniki dotychczas badań przeprowadzonych w celu ustalenia związku między polimorfizmem rs10830963 w genie MTNR1B a ryzykiem cukrzycy ciążowej (gestational diabetes mellitus, GDM) nie pozwoliły na sformułowanie jednoznacznych wniosków. Niniejsze badanie przeprowadzono w celu wyjaśnienia tych kontrowersji, zwłaszcza w odniesieniu do występowania tych związków w populacji chińskiej. Materiał i metody: W wyniku przeszukania w sposób systematyczny piśmiennictwa obejmującego okres do 12 sierpnia 2016 roku wytypowano 35 artykułów, spośród których 11 badań spełniało kryteria włączenia do metaanalizy. Obejmowały one 3889 chorych z GDM i 6708 osób kontrolnych. Wyniki: Autorzy stwierdzili statystycznie istotny związek między polimorfizmem rs10830963 a GDM, obliczając ilorazy szans (odds ratio, OR) i 95-procentowe przedziały ufności (confidence interval, CI) [genotyp GG vs. genotyp CC: OR = 1,70; 95% CI: 1,38–2,10; allel G vs. allel C: OR = 1,27; 95% CI: 1,20–1,36; GG+CG vs CC (model dominujący): OR = 1,31; 95% CI: 1,20–1,44; GG vs. CG+CC (model recesywny): OR = 1,41; 95% CI: 1,26–1,58]. W analizach podgrup wydzielonych na podstawie pochodzenia etnicznego również stwierdzono, że polimorfizm rs10830963 wiąże się z istotnie wyższym ryzykiem GDM we wszystkich modelach genetycznych w populacji chińskiej. Wnioski: Przeprowadzona przez autorów metaanaliza wskazuje, że polimorfizm rs10830963 może być uważany za czynnik ryzyka GDM w populacji chińskiej

Fluorescence-activated cell sorting and directed evolution of α-N-acetylgalactosaminidases using a quenched activity-based probe (qABP)

10.1039/C3CC42836BChemical Communications49657237-723

Razvoj kvantitativnog PCR testa temeljenog na SYBR Green I za identifikaciju cirkovirusa svinja 1, 2 i 3

Porcine Circovirus (PCV) includes Porcine Circovirus 1(PCV1), Porcine Circovirus 2 (PCV2) and Porcine Circovirus 3 (PCV3). In recent years, co-infection exists between PCV1, PCV2 and PCV3 serotypes. Therefore, it is particularly necessary to establish a fast, specific and sensitive SYBR Green I real-time quantitative PCR detection method for PCV1, PCV2 and PCV3. In this experiment, specific primers were selected and the reaction conditions were optimized. A real-time quantitative PCR identification method was established. The results showed the detection limits of this assay were 40.3 copies/μl for PCV1, 25.2 copies/μl for PCV2 and22.4 copies/ μl for PCV3. There was no cross-reactivity with swine fever virus (CSFV), porcine reproductive and respiratory syndrome virus (PRRSV), porcine pseudorabies virus (PRV) and porcine parvovirus (PPV). The intra-assay and inter-assay coefficients of variation were less than 1%. The test results of 100 PCV suspected positive samples revealed that the PCV1, PCV2 and PCV3 singular infection rate was 10% (10/100), 64% (64/100) and 52% (52/100), respectively. The PCV1 and PCV2 co-infection rate was 8% (8/100), the PCV1 and PCV3 co-infection rate was 7% (7/100), the PCV2 and PCV3 co-infection rate was 26% (26/100), and the PCV1, PCV2 and PCV3 co-infection rate was 7% (7/100). This method has good specificity, sensitivity and stability. It provides a promising tool for rapid differential detection of PCV1, PCV2 and PCV3.Među cirkovirusima svinja razlikujemo cirkovirus svinja tipa 1 (PCV1), cirkovirus svinja tipa 2 (PCV2) i cirkovirus svinja tipa 3 (PCV3). Posljednjih se godina pojavljuje koinfekcija ovim trima serotipovima, stoga je potrebno uspostaviti brzu, specifičnu i osjetljivu metodu kako bi se kvantitativnim PCR testom temeljenom na SYBR Green I mogli identificirati PCV1, PCV2 i PCV3. U ovom su istraživanju upotrijebljene specifične početnice te su optimizirani uvjeti rekacije za uspostavljanje kvantitativnog PCR-a u stvarnom vremenu. Rezultati su pokazali da su granice detekcije ovog testa 40,3 kopije/μL za PCV1, 25,2 kopije/μL za PCV2 i 22,4 kopije/μL za PCV3. Nije bilo križne reaktivnosti s virusom svinjske kuge (CSFV), virusom reproduktivnog i respiratornog sindroma svinja (PRRSV), virusom pseudobjesnoće svinja (PRV) i parvovirusom svinja (PPV). Koeficijenti varijacije unutar testa i među testovima bili su manji od 1 %. Rezultati analize 100 uzoraka sa sumnjom na PCV pokazali su da je stopa infekcije serotipom PCV1 bila 10% (10/100), PCV2 64% (64/100), a PCV3 52% (52/100). Stopa koinfekcije serotipovima PCV1 i PCV2 bila je 8% (8/100), PCV1 i PCV3 7% (7/100), a PCV2 i PCV3 26% (26/100). Koinfekcija svim trima serotipovima, PCV1, PCV2 i PCV3, bila je 7% (7/100). Metoda primjenjena u ovom istraživanju ima dobru specifičnost, osjetljivost i postojanost te je obećavajući alat za brzo otkrivanje serotipova PCV1, PCV2 i PCV3

Hand-drawn sketch and vector map matching based on topological features

In the process of addressing, when people use words to express indistinctly, they often draw simple sketches to assist expression, which helps people to form a simple spatial scene in the brain and correspond to the actual scene one by one, and finally locate and find the target address. How to establish an one-to-one mapping relationship between the spatial objects in the hand-drawn sketch and in the vector map is the key to the realization of map addressing and location, and this process is also the process of map matching. This paper aims to address difficult problems associated with the features of hand-drawn sketches and vector map matching in order to improve the use of all potential matching points designed for application in hand-drawn sketches and spatial relation matrix structures of vector maps. To accomplish this, we use the N-queen problem solving process and improve the tabu search algorithm. In the matching process under the constraint of a single spatial relationship, and the hierarchical matching process under the constraint of multiple spatial relations, this study verifies the quality of the spatial relationship and the feasibility and effectiveness of the matching method of hand-drawn sketches and vector maps using the improved tabu search algorithm

Suppress HBV by therapeutic vaccine

乙肝预防性疫苗显著减少了乙肝新发感染，但目前全球仍有约2.5亿慢性乙肝感染者，若未得到有效治疗，可能发展为肝癌、肝硬化等终末期肝病并导致死亡。夏宁邵教授团队研究发展了一种新型的B细胞表位嵌合型类病毒颗粒乙肝治疗性疫苗（治疗性蛋白），在多种模型中证实了其对慢性乙肝感染的治疗潜力，为研发治疗慢性乙肝的原创药物提供了新思路。 我校博士后张天英、博士生郭雪染和博士生巫洋涛为该论文共同第一作者，夏宁邵教授、袁权副教授、张军教授为该论文的共同通讯作者。【Abstract】Objective: This study aimed to develop a novel therapeutic vaccine based on a unique B cell epitope and investigate its therapeutic potential against chronic hepatitis B (CHB) in animal models. Methods: A series of peptides and carrier proteins were evaluated in HBV-tolerant mice to obtain an optimized therapeutic molecule. The immunogenicity,therapeutic efficacy and mechanism of the candidate were investigated systematically. Results: Among the HBsAg-aa119-125-containing peptides evaluated in this study, HBsAg-aa113-135 (SEQ13) exhibited the most striking therapeutic effects. A novel immuno-enhanced virus-like particle carrier (CR-T3) derived from the roundleaf bat HBV core antigen (RBHBcAg) was created and used to display SEQ13, forming candidate molecule CR-T3-SEQ13. Multiple copies of SEQ13 displayed on the surface of this particulate antigen promote the induction of a potent anti-HBs antibody response in mice, rabbits and cynomolgus monkeys. Sera and purified polyclonal IgG from the immunized animals neutralized HBV infection in vitro and mediated efficient HBV/HBsAg clearance in the mice. CR-T3-SEQ13-based vaccination induced long-term suppression of HBsAg and HBV DNA in HBV transgenic mice and eradicated the virus completely in hydrodynamic-based HBV carrier mice. The suppressive effects on HBsAg were strongly correlated with the anti-HBs level after vaccination, suggesting that the main mechanism of CR-T3-SEQ13 vaccination therapy was the induction of a SEQ13-specific antibody response that mediated HBV/HBsAg clearance. Conclusions: The novel particulate protein CR-T3-SEQ13 suppressed HBsAg effectively through induction of a humoral immune response in HBV-tolerant mice. This B cell epitope-based therapeutic vaccine may provide a novel immunotherapeutic agent against chronic HBV infection in humans.This work was supported by the National Scientific and Technological Major project (2017ZX10202203-001), the National Natural Science Foundation of China (31730029, 81672023, 81871316 and 81702006) and the Xiamen University President Fund Project (20720160063). 该研究获得了“艾滋病和病毒性肝炎等重大传染病防治”科技重大专项、国家自然科学基金等资助

The deubiquitinase USP6 affects memory and synaptic plasticity through modulating NMDA receptor stability

人类与其他动物相比的重要区别在于人类拥有高等认知能力，这种能力集中体现在学习记忆和语言表达方面。厦门大学医学院神经科学研究所王鑫教授团队发现人科动物特异性基因USP6作为一个新的NMDA受体调控因子，可通过去泛素化途径调节NMDA型谷氨酸受体的降解和稳定性，进而调控突触可塑性和学习记忆能力。 本研究工作由王鑫教授指导完成，博士生曾凡伟、马学海与硕士生朱琳为共同第一作者，王鑫教授为通讯作者。Ubiquitin-specific protease (USP) 6 is a hominoid deubiquitinating enzyme previously implicated in intellectual disability and autism spectrum disorder. Although these findings link USP6 to higher brain function, potential roles for USP6 in cognition have not been investigated. Here, we report that USP6 is highly expressed in induced human neurons and that neuron-specific expression of USP6 enhances learning and memory in a transgenic mouse model. Similarly, USP6 expression regulates N-methyl-D-aspartate-type glutamate receptor (NMDAR)-dependent long-term potentiation and long-term depression in USP6 transgenic mouse hippocampi. Proteomic characterization of transgenic USP6 mouse cortex reveals attenuated NMDAR ubiquitination, with concomitant elevation in NMDAR expression, stability, and cell surface distribution with USP6 overexpression. USP6 positively modulates GluN1 expression in transfected cells, and USP6 down-regulation impedes focal GluN1 distribution at postsynaptic densities and impairs synaptic function in neurons derived from human embryonic stem cells. Together, these results indicate that USP6 enhances NMDAR stability to promote synaptic function and cognition.This work was partially supported by the National Natural Science Foundation of China (31871077, 81822014, 81571176 to XW; 81701349 to Hongfeng Z.; 81701130 to QZ; and 81471160 to HS), the National Key R&D Program of China (2016YFC1305900 to XW and HS), the Natural Science Foundation of Fujian Province of China (2017J06021 to XW), the Fundamental Research Funds for the Chinese Central Universities (20720150061 to XW and 20720180040 to ZS), Open Research Fund of State Key Laboratory of Cellular Stress Biology, Xiamen University (SKLCSB2019KF012 to QZ), and China Postdoctoral Science Foundation (2017M612130 to QZ).该研究得到了国家自然科学基金面上项目和优秀青年基金项目的支持

Meta-Analysis Results on the Association Between TP53 Codon 72 Polymorphism With the Susceptibility to Oral Cancer

Objectives: TP53 is an important tumor suppressor gene to maintain genomic integrity, and its mutations increase the susceptibility to oral carcinoma. Previous published studies have reported the relation of TP53 codon 72 polymorphism with the risk of oral carcinoma, but the results remain controversial and inconclusive.Methods: We therefore utilized meta-analysis based on a comprehensive search in PubMed, EMBASE, and Google of Scholar databases up to August 19, 2017.Results: Total 3,525 cases and 3,712 controls from 21 case-control studies were selected. We found no significant association between TP53 codon 72 polymorphism and oral carcinoma susceptibility in all genetic contrast models, including subgroup analysis based on control source and ethnicity. Furthermore, TP53 codon 72 polymorphism was not significant associated with oral carcinoma susceptibility in tobacco or alcohol use, and HPV infection status. Our results were confirmed by sensitivity analysis and no publication bias was found.Conclusions: Taken together, our data indicate that TP53 codon 72 polymorphism is not associated with the susceptibility to oral carcinoma