Genetically engineered pre-microRNA-34a prodrug suppresses orthotopic osteosarcoma xenograft tumor growth via the induction of apoptosis and cell cycle arrest.

Osteosarcoma (OS) is the most common primary malignant bone tumor in children, and microRNA-34a (miR-34a) replacement therapy represents a new treatment strategy. This study was to define the effectiveness and safety profiles of a novel bioengineered miR-34a prodrug in orthotopic OS xenograft tumor mouse model. Highly purified pre-miR-34a prodrug significantly inhibited the proliferation of human 143B and MG-63 cells in a dose dependent manner and to much greater degrees than controls, which was attributed to induction of apoptosis and G2 cell cycle arrest. Inhibition of OS cell growth and invasion were associated with release of high levels of mature miR-34a from pre-miR-34a prodrug and consequently reduction of protein levels of many miR-34a target genes including SIRT1, BCL2, c-MET, and CDK6. Furthermore, intravenous administration of in vivo-jetPEI formulated miR-34a prodrug significantly reduced OS tumor growth in orthotopic xenograft mouse models. In addition, mouse blood chemistry profiles indicated that therapeutic doses of bioengineered miR-34a prodrug were well tolerated in these animals. The results demonstrated that bioengineered miR-34a prodrug was effective to control OS tumor growth which involved the induction of apoptosis and cell cycle arrest, supporting the development of bioengineered RNAs as a novel class of large molecule therapeutic agents

[μ-3-(8-Quinol­yloxy)propanedi­yl]bis­[dicarbon­yl(η5-methoxy­carbonyl­cyclo­penta­dien­yl)molybdenum(III)]

The crystal structure of the title dimolybdenum complex, [Mo2(C12H9NO)(C7H7O2)2(CO)4], has a quasi-tetra­hedral Mo2C2 cluster core attached to one 3-(8-quinol­yloxy)propanediyl (L) and two methoxy­carbonyl­cyclo­penta­dienyl (Cp’) ligands which are coordinated to two Mo atoms: one Mo atom bonds two terminal CO ligands while the other links one terminal and one semi-bridging CO ligand. An intra­molecular C—H⋯N hydrogen bond results in the quinolyl plane of the L ligand approaching and being nearly perpendicular to one of the Cp’ rings [88.09 (12)°]. In the supra­molecular structure, a one-dimensional comb-shaped infinite chain is formed approximately along the crystallographic c axis by a combination of inter­molecular C—H⋯O hydrogen bonds and locally generates a C(6) motif. Finally, pairs of inversion-related comb-shaped chains associate into a new ladder-shaped infinite chain through weak π–π stacking inter­actions between neighbouring quinolyl systems (pyridyl centroid–centroid distance = 3.853 Å)

Genetic Diversity and Genome-Wide Association Study of Major Ear Quantitative Traits Using High-Density SNPs in Maize

Kernel and ear traits are key components of grain yield in maize (Zea mays L.). Investigation of these traits would help to develop high-yield varieties in maize. Genome-wide association study (GWAS) uses the linkage disequilibrium (LD) in the whole genome to determine the genes affecting certain phenotype. In this study, five ear traits (kernel length and width, ear length and diameter, cob diameter) were investigated across multi-environments for 2 years. Combining with the genotype obtained from Maize SNP50 chip, genetic diversity and association mapping in a set of 292 inbred lines were performed. Results showed that maize lines were clustered into seven subgroups and a total of 20 SNPs were found to be associated with ear traits significantly (P < 3.95E-05). The candidate genes identified by GWAS mainly encoded ubiquitin-activation enzymes (GRMZM2G015287), carotenoid cleavage dioxygenase (GRMZM2G446858), MYB-CC type transfactor, and phosphate starvation response protein 3, and they were associated with kernel length (KL) and ear diameter (ED), respectively. Moreover, two novel genes corresponding to RNA processing and fructose metabolism were found. Further, the SNPs detected by GWAS were confirmed by meta-QTL analysis. These genes and SNPs identified in the study would offer essential information for yield-related genes clone and breeding program in maize

Arbitrarily primed sequence-related amplified polymorphism (AP-SRAP)

Sequence-related amplified polymorphism (SRAP) is a new-type molecular technique that targets coding sequences in the genome and results in a moderate number of co-dominant markers. Based on the SRAP program, the random primer combinations of SRAP, amplified fragment length polymorphism (AFLP) and simple sequence repeat (SSR) were used as new primers in marker analysis. We defined this technique as arbitrarily primed sequence-related amplified polymorphism (AP-SRAP). Of 256 tested AP-SRAP primers, 37.6% primers produced polymorphic patterns from the DNA of one or more species, which showed that AP-SRAP is an effective method to screen markers. Additionally, 80 SRAP primers were used to screen markers in seven plant species (Chinese cabbage, Chinese kale, eggplant, pepper, cucumber, rose and lily), which indicated obvious polymorphism. The primers of AP-SRAP combine simply and reliably. It can overcome the limitation of the number of standard SRAP primers, make greater use of the supply of alternative primers, and potentially reduce laboratory costs. We expect that AP-SRAP may be of wide application in identity testing, population studies, linkage analysis and genome mapping.Keywords: Arbitrarily primed amplification, DNA markers, plantsAfrican Journal of Biotechnology Vol. 12(29), pp. 4588-459

Synthesis of Enantiomers of Chiral Ester Derivatives Containing an Amide Group and their Chiral Recognition by ¹H NMR Spectroscopy

Enantiomers of Chiral Ester Derivatives Containing an Amide Group and Possessing One or Two Stereogenic Centers Were Prepared from L- and D-Α-Amino Acids, and Glycine with (S)- and (R)-Mandelic Acid for Probing their Chiral Recognition as a New Class of Chiral Guests by 1H NMR Spectroscopy, Since Chiral Ester Derivatives Have Been Rarely Used as Chiral Substrates for Chiral Recognition by 1H NMR Technology. the Results Indicated that These Chiral Ester Derivatives Have Been Successfully Differentiated in the Presence of Tetraaza Macrocyclic Chiral Solvating Agents (TAMCSAs) 1 A–1 C. in Order to Better Understand their Chiral Discriminating Behavior, Job Plots, Association Constants (Ka), and Theoretical Calculations of (S,S)-G1 and (R,R)-G1, as a Representative Example, Were Performed, respectively. in Order to Evaluate their Practical Application, the 1H NMR Spectra of G1 and G9 with Various Optical Purities Were Measured (Up to 98 % Ee). in This Work, a Practical Strategy Has Been Effectively Established for Chiral Recognition of Chiral Ester Derivatives Containing an Amide Group and Possessing One or Two Chiral Centers in the Presence of Tetraaza Macrocyclic Chiral Solvating Agents 1 A–1 C by Means of 1H NMR Spectroscopy

3-Benzyl-6-butyl-5-propyl-3H-1,2,3-triazolo[4,5-d]pyrimidin-7(6H)-one

In the title compound, C18H23N5O2, the triazolopyrimidine ring system is essentially planar, with a maximum displacement of 0.032 (2) Å, and forms a dihedral angle of 87.59 (15)° with the phenyl ring. In the crystal, mol­ecules are linked by inter­molecular C—H⋯O hydrogen bonds and C—H⋯π inter­actions into chains parallel to the c axis

Cashmere growth control in Liaoning cashmere goat by ovarian carcinoma immunoreactive antigen-like protein 2 and decorin genes

Objective The study investigated the biological functions and mechanisms for controlling cashmere growth of Liaoning cashmere goat by ovarian carcinoma immunoreactive antigen-like protein 2 (OCIAD2) and decorin (DCN) genes. Methods cDNA library of Liaoning cashmere goat was constructed in early stages. OCIAD2 and DCN genes related to cashmere growth were identified by homology analysis comparison. The expression location of OCIAD2 and DCN genes in primary and secondary hair follicles (SF) was performed using in situ hybridization. The expression of OCIAD2 and DCN genes in primary and SF was performed using real-time polymerase chain reaction (PCR). Results In situ hybridization revealed that OCIAD2 and DCN were expressed in the inner root sheath of Liaoning cashmere goat hair follicles. Real-time quantitative PCR showed that these genes were highly expressed in SF during anagen, while these genes were highly expressed in primary hair follicle in catagen phase. Melatonin (MT) inhibited the expression of OCIAD2 and promoted the expression of DCN. Insulin-like growth factors-1 (IGF-1) inhibited the expression of OCIAD2 and DCN, while fibroblast growth factors 5 (FGF5) promoted the expression of these genes. MT and IGF-1 promoted OCIAD2 synergistically, while MT and FGF5 inhibited the genes simultaneously. MT+IGF-1/MT+FGF5 inhibited DCN gene. RNAi technology showed that OCIAD2 expression was promoted, while that of DCN was inhibited. Conclusion Activation of bone morphogenetic protein (BMP) signaling pathway up-regulated OCIAD2 expression and stimulated SF to control cell proliferation. DCN gene affected hair follicle morphogenesis and periodic changes by promoting transforming growth factor-β (TGF-β) and BMP signaling pathways. OCIAD2 and DCN genes have opposite effects on TGF-β signaling pathway and inhibit each other to affect the hair growth

Effects of RNA interference-mediated gene silencing of JMJD2A on human breast cancer cell line MDA-MB-231 in vitro

Previous data demonstrate that JMJD2A is a cancer-associated gene and may be involved in human breast cancer by demethylation of H3K9me3. The aim of this study was to investigate depressive effects on JMJD2A by transfection with JMJD2A-sepcific siRNA in human breast cancer cell line MDA-MB-231 and effects on cell proliferation, invasion and migration. JMJD2A-specific siRNA was chemically synthesised and transfected into human breast cancer cell line MDA-MB-231. Expression levels of JMJD2A were detected by quantitative real-time PCR and Western blot analysis. Cells proliferation was evaluated by using flow cytometric anlysis and MTT assay. The abilities of invasion and migration were evaluated by cell migration and invasion assay with Boyden chambers. The results showed that the transfection was successful and expression levels of JMJD2A mRNA and protein in siRNA group were both down-regulated. By MTT assay, the mean actual absorbance in siRNA group was significantly lower than that in blank control group (P < 0.05) and negative control group (P < 0.05). In addition, the percentage of cells in G0/G1 phase in siRNA group was significantly more than that in blank control group (P < 0.05) and negative control group (P < 0.05). Furthermore, by cell invasion and migration assay, the decreased number of migrated cells in siRNA group was observed (P < 0.05). These data imply that silencing JMJD2A gene could result in cell cycle change and proliferation inhibition, and lead to suppress tumor cell invasion and migration. It provides a new perspective in understanding the pleiotropic functions of JMJD2A and its contribution to human breast cancer