VP2 mediates the release of the feline calicivirus RNA genome by puncturing the endosome membrane of infected cells.

Feline calicivirus (FCV) is one of the few members of the Caliciviridae family that grows well in cell lines and, therefore, serves as a surrogate to study the biology of other viruses in the family. Conley et al. (14) demonstrated that upon the receptor engagement to the capsid, FCV VP2 forms a portal-like assembly, which might provide a channel for RNA release. However, the process of calicivirus RNA release is not yet fully understood. Our findings suggest that the separation of the FCV capsid from its genome RNA (gRNA) occurs rapidly in the early endosomes of infected cells. Using a liposome model decorated with the FCV cell receptor fJAM-A, we demonstrate that FCV releases its gRNA into the liposomes by penetrating membranes under low pH conditions. Furthermore, we found that VP2, which is rich in hydrophobic residues at its N-terminus, functions as the pore-forming protein. When we substituted the VP2 N-terminal hydrophobic residues, the gRNA release efficacy of the FCV mutants decreased. In conclusion, our results suggest that in the acidic environment of early endosomes, FCV VP2 functions as the pore-forming protein to mediate gRNA release into the cytoplasm of infected cells. This provides insight into the mechanism of calicivirus genome release.IMPORTANCEResearch on the biology and pathogenicity of certain caliciviruses, such as Norovirus and Sapovirus, is hindered by the lack of easy-to-use cell culture system. Feline calicivirus (FCV), which grows effectively in cell lines, is used as a substitute. At present, there is limited understanding of the genome release mechanism in caliciviruses. Our findings suggest that FCV uses VP2 to pierce the endosome membrane for genome release and provide new insights into the calicivirus gRNA release mechanism.

Effect of different bone cement distributions in percutaneous kyphoplasty on clinical outcomes for osteoporotic vertebral compression fractures: A retrospective study.

Osteoporotic fractures and their complications are becoming increasingly harmful to the elderly. This study aimed to evaluate the clinical results of connected or unconnected bilateral cement after bilateral percutaneous kyphoplasty (PKP) in patients with osteoporotic vertebral compression fractures (OVCF). The clinical data of 217 patients with single-segment OVCF were retrospectively collected. Patients were allocated into 2 groups according to the bilateral bone cement in the vertebrae was connected or unconnected after surgery. The surgery-related indexes of the 2 groups were compared, including operation time; bone cement injection volume; contact situation between bone cement and the upper and lower endplates of the vertebral body; visual analogue scale (VAS) scores before surgery, 1 week and 1 year after surgery; Oswestry disability index (ODI) before surgery, 1 week and 1 year after surgery; local kyphosis angle (LKA) before surgery, 1 week and 1 year after surgery; postoperative vertebral body height at 1 week and 1 year after surgery; vertebral body height restoration rate (HRR) at 1 week and 1 year after surgery. The follow-up results of all patients were recorded. The postoperative VAS, ODI, vertebral body height, LKA and other indexes of the 2 groups were significantly improved compared with those before the operation (P < .05), and there was no significant difference between the 2 groups (P > .05). At the same time, there were no significant difference in vertebral body HRR and bone cement leakage rate between the 2 groups (P > .05). X-ray examination showed that 21 of 217 patients (21/217, 9.8%) had a refracture of the injured vertebral body, including 16 cases (16/121, 13.2%) in the unconnected group and 5 cases (5/96, 5.2%) in the connected group (P < .05). Adjacent vertebrae fractures occurred in 25 cases (25/217, 11.5%), while 19 cases (19/121, 15.7%) were in the unconnected group and 6 cases (6/96, 6.3%) were in the connected group (P < .05). PKP has a good therapeutic effect on OVCF no matter whether the bilateral bone cement is connected or not. However, if the bilateral cement inside the vertebra was connected, the risk of recollapse of the injured vertebrae and the new fracture of adjacent vertebrae could be reduced.

Prefabrication of vascularized porous three-dimensional scaffold induced from rhVEGF(165): a preliminary study in rats.

BACKGROUND/AIMS: Several shortcomings have limited the routine use of autogenous vascularized bone graft. The present study investigates the prefabrication of vascularized scaffold with the desired shape and microarchitecture combined with recombinant human vascular endothelial growth factor 165 (rhVEGF(165)) to mimic autogenous vascularized bone graft. METHODS: Eighty-five porous calcium phosphate cement scaffolds constructed by rapid prototyping technology were divided into four groups: group A [rhVEGF(165)-fibrin sealant (FS) scaffold], group B (hVEGF(165) scaffold), group C (FS scaffold), and group D (scaffold alone). The release of rhVEGF(165) from the scaffolds was examined in vitro. The vessel density, relative functionalized vessels, vessel diameter and relative vessel area were also measured. RESULTS: The sustained release of hVEGF(165) lasted 14 days in the absence of plasmin and 12 days in the presence of plasmin in group A and 10 days in group B. There was no statistical difference between groups A and B at 2 or 4 weeks in terms of vessel density, relative functionalized vessels, vessel diameter, and relative vessel area, as between groups C and D. However, the above parameters were greater in groups A and B than groups C and D. CONCLUSION: The scaffolds with the desired shape and microarchitecture combined with rhVEGF(165) could shorten the time needed for the construction of prefabricated vascularized grafts and accelerate the maturation of the vessels.

Effect of one versus two distal locking bolts on the biomechanics of tibial interlocking nail: a comparative study using a new model.

OBJECTIVE: To investigate the impact of one versus two distal locking bolts on the mechanical properties of tibial interlocking intramedullary nails. METHODS: Twenty 9-mm titanium alloy locking nails were divided into two equal groups in which the nails were fixed with only one and two distal locking bolts, respectively. Each group was further divided into two sub-groups for compression and torsion tests separately using a universal material-testing machine. RESULTS: In the compression tests, the average maximum strength of double bolts was greater than that of the single bolt (P<0.05), but the number of the bolts did not significantly affect the results of the torsion tests in terms of the either the maximum torsion moment or angle (P>0.05). CONCLUSION: One distal bolt is sufficient for fixing stable fractures and double bolts are recommended for management of serious fractures. The model we used is convenient and economic for examining the biomechanics of the tibial nails, especially for comparative purposes.

Osteogenesis of the construct combined BMSCs with beta-TCP in rat.

BACKGROUND: The use of artificial bone graft substitutes has increased as the surgical applications widen and the availability of allograft bone decreases. The present study was to evaluate the construct combined bone marrow stromal cells (BMSCs) with beta-tricalcium phosphate (beta-TCP) as bone substitute implanted in rat dorsal muscles. METHODS: To study the osteogenic capability in vivo, specimens were harvested on 1 week, 4 weeks and 8 weeks after implantation, and were analyzed by hematoxylin and eosin (HE) staining. The percentages of new bone formation for each implant type and implantation period were determined by histomorphometry. RESULTS: After 1 week of implantation, new bone formation for both beta-TCP and BMSCs+beta-TCP group had no formed. After 4 weeks of implantation, the amount of bone formation was increased to 1.32 % in beta-TCP group and 6.35% in BMSCs+beta-TCP group. After 8 weeks of implantation, more bone was found in the BMSCs+beta-TCP group (21.58 %), while in the beta-TCP group bone formation was increased to 4.78%. Significant differences between the two groups have been observed. CONCLUSIONS: Based on these results, we conclude that bone substitutes constructed by porous beta-TCP scaffold loaded with osteogenically induced BMSCs could promote newly formed bone.