ObjectiveTo compare the effects of the Rho-associated protein kinase (ROCK) inhibitors Fasudil and Y27632 on promoting angiogenesis in vivo and in vitro and on flap survival in mice. Methods Thirty-six SPF-grade adult male ICR mice were randomly divided into the Y27632 group, Fasudil group, and control group, with 12 mice in each group. From 3 d before surgery, the mice were intraperitoneally injected daily with Y27632 solution, Fasudil solution, and an equal volume of normal saline, respectively. Then, a peninsular flap model pedicled on the iliolumbar vessels on the back of the mice was established surgically. Laser speckle contrast imaging was used to evaluate flap blood perfusion immediately after surgery (day 0) and on day 5, and the pedicle artery diameter was measured and proximal and distal blood flow were calculated. On day 5 after surgery, the flap necrosis rate was calculated, and CD31-positive microvessel density in the distal part of the flap was detected by immunofluorescence. In addition, EdU cell proliferation, scratch, Transwell, and Matrigel tube formation assays were used to compare the effects of Fasudil and Y27632 on the proliferation, migration, invasion, and tube formation ability of human umbilical vein endothelial cells (HUVECs) in vitro, with DMSO treatment as the control. ResultsOn day 5 after surgery, the flap necrosis rate in the Fasudil group was (2.0±1.5)%, which was significantly lower than that in the control group and the Y27632 group (P<0.001), and the necrosis rate in the Y27632 group was also significantly lower than that in the control group (P<0.001). The pedicle iliolumbar artery diameter in the Fasudil group on day 0 and day 5 after surgery was (143.8±4.2) μm and (177.6±7.6) μm, respectively, both significantly larger than those in the control group and the Y27632 group (P<0.001). On day 5 after surgery, the proximal and distal blood flow in the Fasudil group were (453.0±12.5) PU and (79.2±4.8) PU, respectively, both significantly higher than those in the control group(P<0.001) and the Y27632 group (P<0.05). The microvessel density in the distal surviving tissue of the flap was (71.6±3.5) vessels/region, significantly higher than that in the control group (P<0.001) and also significantly higher than that in the Y27632 group (P＜0.05). On day 0 after surgery, the pedicle iliolumbar artery diameter in the Y27632 group was smaller than that in the control group (P＜0.05), while on day 5 after surgery, there was no statistically significant difference between the two groups (P>0.05). The proximal and distal blood flow on day 5 after surgery were also significantly higher than that in the control group (P<0.001), and the microvessel density was also significantly higher than that in the control group (P<0.05), but the distal blood flow showed no significant difference compared with the control group (P>0.05). In vitro experiments showed that the proliferation rate, migration rate, and number of invasive cells of HUVECs in the Fasudil and Y27632 groups were significantly increased compared with the control group (P<0.001), but there was no statistically significant difference between the two drug groups (P>0.05), whereas the tube formation ability of the Fasudil group was significantly better than that of the Y27632 group (P<0.05). ConclusionBoth Fasudil and Y27632 can effectively promote angiogenesis and improve flap survival rate, but the effect of Fasudil is more significant, which may be related to its sustained regulation of hemodynamics and vascular stability.

Objective: To investigate the brain volume alterations in patients with hyposmia using voxel-based morphometry (VBM) and to correlate these alterations with the degree and duration of hyposmia. Methods: Forty patients with hyposmia from Department of Otorhinolaryngology Head and Neck Surgery, Beijing Anzhen Hospital since 2013 to 2016 and forty age and sex matched normal subjects were recruited in this study. Sniffin′ Sticks olfactory test was performed to evaluate the olfactory function of all subjects. We acquired T1-weighted magnetic resonance images from all subject on a 3T scanner. VBM was performed using VBM8 toolbox and SPM8 in a Matlab environment. Independent sample t test analysis was used to compare the volume of gray and white matter between the controls and patients. In addition, the Pearson or Spearman correlation analysis was used for the correlation between the voxel value of cerebral volume alterations area and the degree and duration of hyposmia on patients (threshold discrimination identification, TDI). Results: Compared with the controls, patients showed significantly decreased volume in the gray matter of right orbitofrontal cortex (number of voxel in clusters was 226, t=-4.46, P<0.001, uncorrected). There was negative correlation between decreased gray matter volume of the right orbitofrontal cortex with significantly decreased area and the TDI results (r=-0.40, P=0.01), but positive correlation with duration of hyposmia (r=0.37, P=0.02). Conclusions The patients with hyposmia show gray matter atrophy of the right orbitofrontal cortex. The duration may be an important risk factor for decreased gray matter in patients with hyposmia.