Biochemical Genetics

Phosphorylase kinase deficiency in I strain mice and in humans both show X-chromosomal inheritance. Neither deficient adult humans nor deficient mice show any sign of disease. Thus the two conditions resemble each other. However, there are differences. The enzyme is only partially deficient in human patients in liver, muscle, and blood cells; in mice the deficiency is complete and seems to be confined to the muscles.

Retinoblastoma (Rb) is the most common pediatric malignant tumor of the eyes. Previous studies demonstrated that miR-491-3p is downregulated in various cancers. However, its function in Rb remains unknown. A total of 15 pairs of primary Rb tissues and adjacent noncancerous tissues were collected. Quantitative real-time PCR (qRT-PCR) was used to investigate the expression profiles of miR-491-3p. qRT-PCR, western blotting and in situ immunocytochemistry were performed to investigate the expression profiles of epithelial–mesenchymal transition-related proteins (E-cadherin, Vimentin and N-cadherin) in Rb tissues and Rb cell lines as well as cell morphology. Cell proliferation was estimated by MTS and colony formation assays. Apoptosis was determined by FACS, cell migration and invasion were analyzed using transwell chambers. MiR-491-3p’s target genes were predicted using target gene prediction databases. The interplay between miR-491-3p and SNN was evaluated through dual luciferase reporter gene assay. MiR-491-3p was significantly downregulated in mixed collection of 15 pairs of Rb tissues and Rb cell lines. Overexpression of miR-491-3p enhanced apoptosis, and significantly suppressed proliferation, migration and invasion of Rb cells. In contrast, the present of miR-491-3p inhibitor showed reversed results which apoptosis decreased, while cell proliferation of ARPE-19 cells increased. In addition, miR-491-3p increased the expression of E-cadherin, and dramatically decreased the expression of Vimentin and N-cadherin in Rb tissues and Rb cell lines, noticeable changes in morphology, too, as cells became less cohesive and more adhering. We found out that SNN was the pairing target of miR-491-3p and result showed that miR-491-3p and SNN interacted with each other. We also found out that the effects of miR-491-3p were in Rb cells were almost entirely canceled out at the overexpression of SNN. Our findings collectively suggest that miR-491-3p is an important tumor suppressor in Rb, which inhibits tumor growth and metastasis in Rb. These implicate it may be explored as a new therapeutic target in Rb.

Starch gel electrophoresis of kidney catalase in inbred strains C3H and C57BL/6, their F1 hybrid, and first and second backcross generations demonstrated that single-component (type A) v. multiple-component (type B) electrophoretic patterns are controlled by a single locus. The type A electrophoretic pattern is dominant. Twenty-five inbred strains of mice were classified according to their kidney catalase electrophoretic pattern. The data indicate that the segregating genetic factor determines a specific substance in the type A kidney which affects the electrophoretic mobility of catalase. A comparison of the F1 hybrid enzyme with a 1:1 mixture of C3H and C57BL/6 enzyme showed that the alteration of electrophoretic mobility is the result of posttranslational modification of the catalase molecule. An association of kidney catalase electrophoretic pattern and the H-2 k haplotype indicates that the locus controlling the electrophoretic pattern is most likely located on chromosome 17 in close proximity to the H-2 complex.

The first 539 bases of mitochondrial DNA D-loop region of six Chinese native chicken breeds (Gallus gallus domesticus) were sequenced and compared to those of the red junglefowl (Gallus gallus), the gray junglefowl (Gallus sonneratii), the green junglefowl (Gallus varius) and Lafayette's junglefowl (Gallus lafayettei) reported in GenBank, and the phylogenetic trees for the chickens were constructed based on the D-loop sequences. The results showed that the four species of the genus Gallus had great differences among each other, the G. g. domesticus was closest to the red junglefowl in Thailand and its adjacent regions, suggesting the Chinese domestic fowl probably originated from the red junglefowl in these regions. The two subs pecies of Thailand, G. g. gallus and G. g. spadiceus, should belong to one subspecies because of their resemblance. In the case of native breeds, there existed a great difference between the egg breeds and general purpose breeds, which suggested different maternal origins of the two types.

To clarify whether genetic polymorphisms in exon 14 of Coagulation factor XIII A-subunit gene (FXIIIA) affect phenotype expressions, we studied genetic polymorphisms in exon 14 of FXIIIA in a Japanese population and the relationship between the genetic polymorphisms and phenotype expression. Genetic polymorphisms in exon 14 of FXIIIA of 144 unrelated Japanese were analyzed by single-strand conformation polymorphism. Plasma FXIIIA antigen concentrations, FXIII activities, and phenotype were also determined by two-dimensional electrophoresis. The frequencies of the three genotypes, the homozygote (AD), the homozygote (BC) and the heterozygote (AD/BC), were 77.1, 0.7, and 22.2%, respectively. The gene frequencies of AD and BC were 0.88 and 0.12. We detected AD (GTTand BC (ATTat codon 650 and 651 of exon 14. There were no significant differences of FXIIIA antigen concentrations and FXIII activities between these genotypes. We detected three pl differences among them as being pls of 5.3, 5.6, 5.8 in the homozygote (AD) and the heterozygote (AD/BC), and a pl of 5.8 in the homozygote (BC). These polymorphisms affected isoelectric mobility, but did not affect protein levels, enzyme activities, or the molecular weight of FXIII.

Circular RNAs function as important regulators in the pathogenesis of human cancers, including nasopharyngeal carcinoma (NPC). We aimed to explore the functions of circ_0028007 in NPC development. Quantitative real-time polymerase chain reaction assay was employed for the levels of circ_0028007, NUAK family kinase 1, microRNA-656-3p (miR-656-3p), and E74 like ETS transcription factor 2 (ELF2). RNase R assay was used to verify the feature of circ_0028007. Cell Counting Kit-8 assay and colony formation assay were performed to assess cell growth. Wound-healing assay and transwell assay were adopted to analyze cell migration and invasion. Tube formation assay was conducted for cell angiogenic capacity. Flow cytometry analysis was performed for cell apoptosis. Western blot assay was conducted for protein levels. Compared to normal tissues and cells, circ_0028007 level was elevated in NPC tissues and cells. Knockdown of circ_0028007 repressed NPC cell growth, migration, invasion, and angiogenesis, facilitated apoptosis in vitro and blocked tumor growth in vivo. Moreover, circ_0028007 silencing could regulate the AMP-activated protein kinase/mammalian target of rapamycin pathway in NPC cells. Circ_0028007 promoted the malignant behaviors of NPC cells via acting as miR-656-3p sponge. In addition, ELF2 was demonstrated to be the target gene of miR-656-3p. MiR-656-3p overexpression restrained NPC cell malignant phenotypes, while ELF2 elevation reversed the effects. Circ_0028007 contributed to the progression of NPC by decoying miR-656-3p and elevating ELF2. The findings might provide potential targets for NPC therapy.

The titi monkeys (Callicebus spp.) are a large, diverse genus of platyrrhines, widely distributed in tropical South America. The genetic variability of these monkeys is still relatively poorly known, especially at the population level. In the present study, four heterologous microsatellite markers were used to investigate genetic diversity in 23 individuals from a wild population of red-bellied titis (Callicebus moloch) in eastern Amazonia. An unexpectedly low level of diversity was found. The average number of alleles was 8.75 (range: 5–15), and the average heterozygosity was 0.33 (range: 0.09–0.65). This preliminary information suggests a reduction of the potential for long-term survival of the population and indicates the putative necessity of implementation of a species conservation program.