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 explore the characteristics and advantages of Whole-Mounting immnunofluorescent staining and laser speckle flow imaging technology in the vascular imaging of mouse ear extended flap.Methods:In this study, total of 25 ICR mice were included.Ten ICR mice were cut off the middle and lateral angiosome to establish an extended flap model, and 3 days later, the changes in the blood supply of the ear flap were observed. The ear area , tissue layer thickness and blood vessel distribution in the healthy side were observed at the same time.Obtain the mouse ears 3 days after modeling, and dissect them into three layers, i.e, the anterior skin layer , the cartilage layer and the posterior skin layer. The distribution and morphology of blood vessels, nerves and monocytes/macrophages in the anterior skin layer were stained and detected by the whole-mount immunofluorescence staining.Ten mice were adopted and an incision was made through the ear horizontally above the bifurcation of the middle angiosome of the mouse ear to establish a delayed extended flap model. Then the blood flow changes in the mouse ear were observed by laser speckle flow imaging andt he blood perfusion values were recorded immediately, 1 d, 2 d, 3 d and 4 d after the operation, respectively.Results:The area of the mouse ear was about 1.3 cm 2 , the thickness was about (0.16±0.04) mm, and the blood was supplied by three vascular bundles: the lateral caudal vascular bundle, the middle vascular bundle and the medial cephalic vascular bundle. The thickness of the anterior and posterior skin and cartilage of the mouse ears were (88±5)μm, (41±3)μm and (29±2)μm, respectively. The whole-mount immunofluorescence staining results clearly showed that the diameter of small vessels in the choke area was (50 ± 6) μm on the third day after modeling. It could be seen that the nerve and artery in mouse ear were in concomitant relationship and the nerve segment attached to the surface of the artery without obvious accompany or clinging to the vein. There were a large number of monocyte macrophages distributed in clusters in the dilated and curved arteries, but they were only scattered outside the artery. Laser speckle flow imaging results showed that there were (6 ± 2) transverse vessels in each auricular flap, and the diameter and blood flow increased significantly in the delayed extended earflap model. Immediately after the operation and at 1d, 2d, 3d, and 4d, the average blood perfusion values of transverse vessels were (92±11) PU, (136±26) PU, (147±27) PU and (176±27) PU, respectively. Conclusions:The Whole-Mounting immnunofluorescent staining and the laser speckle blood flow imaging technology can be used to well observe the blood vessels, nerves, mononuclear macrophages and blood flow perfusion of the mouse extended flap, which can play an important role in the study of blood supply of mouse extended flap.

Objective:To explore the characteristics and advantages of Whole-Mounting immnunofluorescent staining and laser speckle flow imaging technology in the vascular imaging of mouse ear extended flap.Methods:In this study, total of 25 ICR mice were included.Ten ICR mice were cut off the middle and lateral angiosome to establish an extended flap model, and 3 days later, the changes in the blood supply of the ear flap were observed. The ear area , tissue layer thickness and blood vessel distribution in the healthy side were observed at the same time.Obtain the mouse ears 3 days after modeling, and dissect them into three layers, i.e, the anterior skin layer , the cartilage layer and the posterior skin layer. The distribution and morphology of blood vessels, nerves and monocytes/macrophages in the anterior skin layer were stained and detected by the whole-mount immunofluorescence staining.Ten mice were adopted and an incision was made through the ear horizontally above the bifurcation of the middle angiosome of the mouse ear to establish a delayed extended flap model. Then the blood flow changes in the mouse ear were observed by laser speckle flow imaging andt he blood perfusion values were recorded immediately, 1 d, 2 d, 3 d and 4 d after the operation, respectively.Results:The area of the mouse ear was about 1.3 cm 2 , the thickness was about (0.16±0.04) mm, and the blood was supplied by three vascular bundles: the lateral caudal vascular bundle, the middle vascular bundle and the medial cephalic vascular bundle. The thickness of the anterior and posterior skin and cartilage of the mouse ears were (88±5)μm, (41±3)μm and (29±2)μm, respectively. The whole-mount immunofluorescence staining results clearly showed that the diameter of small vessels in the choke area was (50 ± 6) μm on the third day after modeling. It could be seen that the nerve and artery in mouse ear were in concomitant relationship and the nerve segment attached to the surface of the artery without obvious accompany or clinging to the vein. There were a large number of monocyte macrophages distributed in clusters in the dilated and curved arteries, but they were only scattered outside the artery. Laser speckle flow imaging results showed that there were (6 ± 2) transverse vessels in each auricular flap, and the diameter and blood flow increased significantly in the delayed extended earflap model. Immediately after the operation and at 1d, 2d, 3d, and 4d, the average blood perfusion values of transverse vessels were (92±11) PU, (136±26) PU, (147±27) PU and (176±27) PU, respectively. Conclusions:The Whole-Mounting immnunofluorescent staining and the laser speckle blood flow imaging technology can be used to well observe the blood vessels, nerves, mononuclear macrophages and blood flow perfusion of the mouse extended flap, which can play an important role in the study of blood supply of mouse extended flap.

Objective: To compare the efficacy of percutaneous transluminal angioplasty (PTA) and neo-anastomosis in juxta-anastomotic stenosis of autologous arteriovenous fistula (AVF) of diabetic hemodialysis patients. Methods: Diabetic patients with juxta-anastomotic stenosis in Beijing Tsinghua Changgung Hospital from February 2016 to December 2018 were retrospectively analyzed. The clinical data, success rate, patency time of PTA and vascular neo-anastomosis were compared. Results: Thirteen patients received PTA, including 1 case of surgical failure and 1 case of thrombosis, and the success rate was 11/13. Vascular neo-anastomosis were performed in 15 cases, and 13 cases succeeded. The success rate was 13/15, and the success rate between two groups had no significant (P > 0.05). The median patency time was 11(7-12) months and 15(5-22) months (P > 0.05). The volume of bleeding [3(2-4) ml vs. 20(10-40) ml], the length of blood vessel wastage [0 vs. (3.6 ± 1.2) cm] and the operating time [(0.6 ± 0.7) h vs. (1.8 ± 0.5) h] of PTA patients were less than those of vascular neo-anastomosis group, and there were significant differences (P < 0.05). Conclusions There are no significant differences between PTA and neo-anastomosis with respect to patency. PTA has the advantages of less vascular injury, shorter operation time, less bleeding and can dilate arteries at the same time.

BACKGROUND:Artificial knee joint replacement is the main surgical method for the treatment of severe knee joint deformation, but due to the limited existing knee prosthesis type. The stature of Fujian population in southeast China was shorter than that of the north. The size of artificial knee joint usual y did not fit for patients in clinic. The substitution effect was poor.
OBJECTIVE:To measure the geometric parameters of proximal surface and cross-section of adult female tibia in southeastern area of Fujian Province of China, and provide basic data for design and replacement of artificial knee joint in females.
METHODS:We chose 85 cases of dry female skeleton in southeastern area of Fujian Province of China, measured tibial plateau width, medial tibial plateau width and anteroposterior diameter, lateral tibial plateau width and anteroposterior diameter. We col ected clinical normal proximal tibial CT data of 36 patients (38-65 years old), with scan range of 30 cm (intercondylar eminence 30 cm) and layer thickness of 0.625 mm. Mimics 16.0 three-dimensional reconstruction software was used to measure above indexes and cut the reconstructed models with thickness of 3 mm, total y 10 layers. We divided its widest distance into four equal parts, and measured tibial anteroposterior diameter on three points. Statistical software was used to analyze the linear relationship between the data.
RESULTS AND CONCLUSION:The experiment has successful y obtained geometric parameters of tibial specimens of adult females in southeastern area of Fujian province of China. Tibia platform inside and outside diameters were greater than the width inside and outside;inside and outside diameter and inside and outside width were similar. Ten segment planes of tibia, inside and outside diameters were always less than middle anteroposterior diameter. Three groups of data increased firstly, and then reduced. There were positive correlations between tibia length and width of the medial tibia platform, the lateral tibia platform width and anteroposterior diameter, the medial tibia platform width and anteroposterior diameter, and the lateral tibia platform width and anteroposterior diameter (P<0.05). These results suggested that relevant parameters of tibial plateau adult women in southeastern area of Fujian province of China showed some regular changes. Our results may provide quantitative reference data for the design and operation of artificial tibial plateau.

Objective To establish a mouse ’ s cross-territory ear flap that enables chronic , in-vivo observation of the change of vascular morphology .Methods 30 ICR mice, weighing 25~40 g, were used for this study .Commercial depilatory cream was used to first remove the hair of the mice , after which the vascular pattern in the ears was investigated . According to the observation of the vascular pattern in the mouse ’ s ear, the eye scissors were used the sever the outer 2/3 of the base of the ear , in which process a ear ’ s flap based on a vascular pedicle but crossed three vascular territories was created.After the creation of the flap , the mice were placed on an automatic controlled movable machine with the ear ’ s flaps spread over a customized Plexiglas .Then the flaps were photographed under the stereoscope ( ×25) at the following time points:1,2,3,5,7,10,14,21,30 d.the necrosis of the flap, and the morphological change of the vessels within theflap were analyzed .Results The ICR mouse ’ s ear was supplied three angiosomes , which were respectively named as the cephalic , median and caudal angiosomes from inside out .Five days after the flap’s creating, necrotic rate of(15 ±7)%was developed .The choke vessels between the medial and median angiosomes expanded rapidly in diameter , reaching the plateau 10d after flap creation, resulting the dilated choke veins and arteries at their peak being 3.9 ±0.5 and 3.5 ±0.7, respectively, than their initial sizes.The diameter of the choke veins began to shrink at approximately 10d, stabilizing after 21d.The diameter of the choke arteries plateaued and stabilized at around 10d.Conclusion ①after harvest of extended flap, the dilation of veins seemed to passive , whereas the dilation of arteries seemed to active;②the number of the choke vessels between the dynamic and potential territories that are involved in dilation and extent of the dilation are much smaller than that of the choke vessels between the anatomic and dynamic territories;③the mouse ’ s ear flap is an excellent model of further study of mechanism underlining the dilation of choke vessels and for the screening of vasoactive drugs that augment the survival of the large flap .

ObjectiveTo investigate anatomical characteristics of the medial foot vessels and effects of different vascular pedicle skin flaps in repair of foot and ankle trauma.MethodsThirty adult cadaveric lower limbs were injected with red latex through the popliteal artery and posterior tibial artery to anatomically observe the cutaneous arterial origin,branches,distribution and anastomosis of the medial foot.Then,anterior medial malleolar artery perforator flaps and distally-based medial foot flaps were harvested and used for repairing foot and ankle trauma of 16 patients.Results The origin of cutaneous blood vessels of the medial foot was diversified and mainly included the anterior medial malleolar artery,medial tarsal artery,and arterial arcades anastomosing with anterior posterior branches of the two former arteries and the superficial branches of plantar digital artery and the medial plantar artery.According to distribution area of the anterior medial malleolar artery and the medial tarsal artery,the vascular anatomy of the medial foot skin was classified into three types.Clinically,all the flaps survived.Follow-up ranged from 2 weeks to 20 months,which showed normal color,good shape and good pain and warm sensation of the flaps.ConclusionThe anterior medial malleolar artery perforator flaps present good reconstruction of soft tissue defects around the ankle,whereas the distally-based medial foot flaps present good reconstruction of soft tissue defects of the mid-forefoot.

Background and purpose: Cetuximab-containing regimen has been increasingly applied in the treatment of patients with metastatic colorectal cancer. Simultaneously, the predictive factors of outcome for cetuximab have been thoroughly researched. The aim of this study was to evaluate the relationship between K-ras status and efficacy of cetuximab containing regimen in the treatment of patients with metastatic eolorectal cancer. Methods: Between March 2006 and June 2008, twenty-seven patients with metastatic colorectal cancer were treated with cetuximab in combination with chemotherapy. The K-ras mutation status [wild-type (wt) or mutation (nat)] was examined by polymerase chain reaction (PCR) and direct sequencing. The influence of K-ras mutation status on efficacy of cetuximab-containing regimen was analyzed. Results: For 27 patiants in this cohort, K-ras wt was detected in 55.6% (15/27) cases and K-ras mt in 44.4% (12/27) cases. Statistically significant differences were found between the patients with K-ras wt and K-ras mt in terms of overall response rate (ORR) (66.7% vs 25.0%,P=0.035) and progression-free survival (PFS) (8 months vs 4 months, P=0.0028). However, there was no significant difference in overall survival (OS) (19 months vs 12 months, P>0.05). The most common treatment-related adverse effect was skin reaction, with incidence rate of 80.0% and 66.7% (P>0.05), respectively. No treatment related death was observed. Conclusion: K-ras mutation status is a predictive factor of good efficacy of cetuximab-containing regimen in the treatment of patients with metastatic colorectal cancer. Patients with K-ras wt could benefit from cetuximab in combination with chemotherapy.