Complete chloroplast genome sequencing of five Salix species and its application in the phylogeny and taxonomy of the genus.

In this study, whole chloroplast genomes of five Salix species (S. argyracea, S. dasyclados, S. eriocephala, S. integra 'Hakuro Nishiki', and S. suchowensis) were sequenced. These chloroplast genomes were 155 ,605, 155, 763, 155, 552, 155, 538, and 155 ,550 bp in length, harboring 131 genes (77 unigenes), 37 tRNA genes, 8 rRNA genes, and 86 mRNA genes, respectively. The genes ycf1, psaI, ycf2-2, rpoC2, rpl22, atpF, and ndhF were under positive selection among the 21 Salix species. psaI, ycf2-2, atpF, and ycf1-2 were under positive selection between the tree willow and shrub willow, and rpoC2, rpl22, and ycf1-2 were positively selected among the shrub genomes. The gene rps7 was most variable among the genomes. Phylogenetic analysis of 21 Salix species and Chosenia arbutifolia provide evidence that the cp genome data partially support the relationship with traditional taxonomic concepts in the Flora of China. This chloroplast genome elucidates Salix taxonomy and provides evidence for evolutionary research.

Mutation analysis of the TUBB8 gene in primary infertile women with arrest in oocyte maturation.

Tubulin beta eight class VIII (TUBB8) is a subtype of ß-tubulin that only exists in primates. Mutations in the TUBB8 gene have been proven to cause oocyte maturation arrest. The aim of this study was to identify the new types of mutations in TUBB8. Six women (families) with oocyte maturation arrest and 100 healthy controls were recruited. The sequence of the TUBB8 gene was amplified and analyzed by Sanger sequencing, which revealed a de novo heterozygous variant c.292G > A (p.G98R) of TUBB8 in one affected individual. This TUBB8 variant was absent in the 100 fertile females and was predicted to be highly damaging to the function of the TUBB8 protein by SIFT and PolyPhen-2. This novel variant extends the spectrum of TUBB8 mutations and the presence of a TUBB8 mutation is being considered to be indicative of a poor prognosis for the success of assisted reproductive treatment.

Outcomes of vitrified-warmed cleavage-stage embryo hatching after in vitro laser-assisted zona pellucida thinning in patients.

The aim of the present study was to determine whether the size of the zona pellucida (ZP) thinning area by laser-assisted hatching affected the potential development of vitrified-warmed embryos. A total of 196 vitrified-warmed cleavage-stage embryos (from 49 patients, four sister embryos per patient) were used in the study, i.e., four sister embryos from each patient were randomly assigned to four groups: a control group of embryos that were not zona-manipulated (zona intact, group A); one experimental group of embryos in which a quarter of the zona pellucida was thinned using laser-assisted ZP thinning (group B); a second experimental group of embryos in which half of ZP was thinned (group C); and a third group in which two-thirds of the ZP was thinned (group D). Subsequent blastocyst development was assessed. Microscopy was performed to study the hatching process of the embryos after zona thinning. The blastocyst formation rates were 71.43% in group A, 67.35% in group B, 65.31% in group C, and 51.02% in group D (groups B-D vs. group A, P=0.661, P=0.515, P=0.038, respectively). The rates of complete hatching were 30.61% in group A, 38.78% in group B, 61.22% in group C, and 48.98% in group D (groups B-D vs. group A, P=0.396, P=0.002, P=0.063, respectively). For a subgroup of patients, there was a significant difference in the complete hatching in all the groups for women aged <35 years (P=0.011), and there was a significant difference in the complete hatching in all the groups for secondary infertility women (P=0.022). There was no significant difference in the blastocyst formation rates in the different groups of women aged ≥35 years (P=0.340). In addition, there was no significant difference in the complete hatching in the different groups among women aged ≥35 years (P=0.492). The results of the present study showed that in vitrified-warmed embryo transfers at the cleavage-stage, and the two-thirds zona pellucida thinning group demonstrated a significantly decreased blastocyst formation rate compared with the control group, while the half zona pellucida thinning group demonstrated a significantly increased complete hatching rate compared with the control group, which may have a high value in clinical application.