Objective:To investigate the feasibility of digital reconstruction technology in virtual planning of the free perforator flap of anterior tibial artery (ATA) for reconstruction of soft tissue defects in foot and ankle.Methods:From May 2018 to April 2023, 10 patients, including 7 males and 3 females, with foot or ankle defects were admitted in the Department of Orthopaedics Surgery, 920th Hospital of Joint Logistics Support Force. There were 5 defects in dorsal foot, 3 in plantar foot and 2 in medial malleolus. The sizes of defects ranged from 3.0 cm×2.5 cm to 5.5 cm×4.0 cm, all with exposed bones or tendons. Preoperative CTA scans from aorta abdominalis to feet were performed, and 3D digital models of bones, arteries and skin were reconstructed with Mimics. The most suitable perforators were selected to design the perforator flaps of ATA with the software, then the digital virtual flaps were superimposed onto the surfaces of donor sites and marked under a translucent image by Sina. During the surgery, flaps were harvested according to preoperative digital designs with the size of 3.5 cm×3.0 cm-6.0 cm×4.0 cm. The perforating branches were dissected along the way, and the origin, diameter, course, location and length of the perforators were recorded. The perforating branches of the flaps were anastomosed to the proximal vessels in the recipient sites, and the flaps were sutured to cover the wound. For the 10 donor sites, skin graft was used in 2 donor sites and direct suture were performed on 8 donor sites. After discharge of the patients, scheduled outpatient or online follow-ups were carried out to assess the progress of fracture healing as well as the appearance, texture and colour of flaps, and the recovery of donor sites. Ankle function was evaluated by Maryland foot scoring system.Results:Three-dimensional digital reconstructions of donor sites were successfully performed on all patients, enabling successful design and harvest of the free perforator flaps of ATA. The flaps were able to be used in complete reconstruction of the respective defects and made the anatomical parameters of perforators of the donor sites closely matching with those of preoperative modeling. Follow-up periods ranged from 6 to 19 months, apart from 1 flap experienced partial necrosis at distal endge, and another flap with partial exfoliation after blistering. The rest of 8 flaps were all survived smoothly, with appropriate thickness, aesthetic appearance, good texture and colour. Sensations in both donor sites and dorsal feet were all normal. Seven patients achieved excellent and 3 were good according to Maryland's ankle-foot function score. The donor sites healed well without scar hyperplasia.Conclusion:Digital reconstruction technology enables an accurate identification of perforators as well as individualised design and harvest procedures for perforator flaps of ATA, thereby it facilitates precise reconstructions of small-to-medium-sized defects in foot or ankle. It is a good method for vascular anatomy and flap harvesting.

Objective:To investigate the feasibility of digital reconstruction technology in virtual planning of the free perforator flap of anterior tibial artery (ATA) for reconstruction of soft tissue defects in foot and ankle.Methods:From May 2018 to April 2023, 10 patients, including 7 males and 3 females, with foot or ankle defects were admitted in the Department of Orthopaedics Surgery, 920th Hospital of Joint Logistics Support Force. There were 5 defects in dorsal foot, 3 in plantar foot and 2 in medial malleolus. The sizes of defects ranged from 3.0 cm×2.5 cm to 5.5 cm×4.0 cm, all with exposed bones or tendons. Preoperative CTA scans from aorta abdominalis to feet were performed, and 3D digital models of bones, arteries and skin were reconstructed with Mimics. The most suitable perforators were selected to design the perforator flaps of ATA with the software, then the digital virtual flaps were superimposed onto the surfaces of donor sites and marked under a translucent image by Sina. During the surgery, flaps were harvested according to preoperative digital designs with the size of 3.5 cm×3.0 cm-6.0 cm×4.0 cm. The perforating branches were dissected along the way, and the origin, diameter, course, location and length of the perforators were recorded. The perforating branches of the flaps were anastomosed to the proximal vessels in the recipient sites, and the flaps were sutured to cover the wound. For the 10 donor sites, skin graft was used in 2 donor sites and direct suture were performed on 8 donor sites. After discharge of the patients, scheduled outpatient or online follow-ups were carried out to assess the progress of fracture healing as well as the appearance, texture and colour of flaps, and the recovery of donor sites. Ankle function was evaluated by Maryland foot scoring system.Results:Three-dimensional digital reconstructions of donor sites were successfully performed on all patients, enabling successful design and harvest of the free perforator flaps of ATA. The flaps were able to be used in complete reconstruction of the respective defects and made the anatomical parameters of perforators of the donor sites closely matching with those of preoperative modeling. Follow-up periods ranged from 6 to 19 months, apart from 1 flap experienced partial necrosis at distal endge, and another flap with partial exfoliation after blistering. The rest of 8 flaps were all survived smoothly, with appropriate thickness, aesthetic appearance, good texture and colour. Sensations in both donor sites and dorsal feet were all normal. Seven patients achieved excellent and 3 were good according to Maryland's ankle-foot function score. The donor sites healed well without scar hyperplasia.Conclusion:Digital reconstruction technology enables an accurate identification of perforators as well as individualised design and harvest procedures for perforator flaps of ATA, thereby it facilitates precise reconstructions of small-to-medium-sized defects in foot or ankle. It is a good method for vascular anatomy and flap harvesting.

BACKGROUNDTat-interacting protein 30 (TIP30) has been reported to be a tumor suppressor, with reduced or absent expression in various tumors. However, its role in bladder urothelial cancer (BUC) has not been investigated. Therefore, herein, we investigated the expression of TIP30 protein in BUC and normal bladder mucosa and the clinical significance of TIP30 expression in the prognosis of BUC.

METHODSWe reviewed data from 79 cases of BUC and 15 adjacent tissue samples from 79 patients treated at our institution between 2004 and 2007. TIP30 expression was examined by immunohistochemistry. The relationship between TIP30 expression and tumor stage, histological grade, and survival was analyzed. Differences between groups were evaluated using the t-test or matched-pairs test, and differences in the survival rates were analyzed with the log-rank test.

RESULTSTIP30 protein expression was significantly reduced in BUC tissue (t = -6.91, P < 0.05) compared with normal tissue samples, and in invasive bladder cancer (t = 10.89, P < 0.05) compared with superficial bladder cancer. TIP30 protein expression differed significantly among different differentiated groups classified either according to the World Health Organization (2004, F = 17.48, P < 0.01) or World Health Organization (1973, F = 10.68, P < 0.01). TIP30 protein expression was significantly reduced in high-grade papillary urothelial carcinoma compared with papillary urothelial neoplasm of low malignant potential (P < 0.05) and low-grade papillary urothelial carcinoma (P < 0.05). Meanwhile, TIP30 protein expression was significantly reduced in Grade III BUC, compared with Grade I (P < 0.05) and Grade II (P < 0.05). Patients with low TIP30 expression showed a higher incidence of disease progression than those with high TIP30 expression (t = 2.63, P < 0.05). Kaplan-Meier survival analysis showed a strong positive relationship between TIP30 expression and overall survival (OS) (χ2 = 17.29, P < 0.05).

CONCLUSIONSTIP30 expression was associated with clinical tumor stage in BUC, suggesting that it might play an important role in disease progression. Furthermore, TIP30 might predict postoperative OS. Thus, its evaluation might be useful for predicting prognosis.

Objective: To investigate the effects and related mechanism of bivalirudin on the survival of random skin flap on the back of rat. Methods: Thirty SD rats were divided into bivalirudin group and normal saline group according to the random number table, with 15 rats in each group. The random flap model with size of 9 cm×3 cm was reproduced on the back of rats in two groups. Immediately post injury, rats in bivalirudin group were intraperitoneally injected with 5 mg/mL bivalirudin (0.8 mL/kg), while rats in normal saline group were intraperitoneally injected with normal saline (0.8 mL/kg) once a day. The continuous injection lasted for 7 days. The flap was divided into distal area, middle area and proximal area averagely based on the flap blood supply. On post injury day (PID) 1, 3, and 7, the overall survival of each area of flap was observed with naked eyes. On PID 7, the survival rate of flap was calculated, and then the morphology of skin tissue at the center of the three areas of flap was observed by HE staining, the microvessel density (MVD) of the middle area of flap was calculated, and the expression of vascular endothelial growth factor (VEGF) of the middle area of flap was detected with immunohistochemical staining. Data were processed with t test. Results: (1) On PID 1, flaps of rats in two groups had different degrees of swelling, mainly concentrated in distal area, but there was no obvious necrosis. The middle area and proximal area of flaps in two groups were survived. On PID 3, the necrosis of flaps of rats in two groups was concentrated in the middle area, while the proximal area of flap was still in survival state, and most distal area of flap was necrosis with a little scab. On PID 7, the necrosis of middle area of flaps of rats in two groups was gradually fused, and the survival area of flap of rats in bivalirudin group was larger than that in normal saline group. The distal area of flap was almost necrotic, and the proximal area of flap was almost survived. (2) On PID 7, the survival rate of flap of rats in bivalirudin group was (64±4)%, significantly higher than that in normal saline group [(45±3)%, t=13.49, P<0.01]. (3) On PID 7, the histological morphology of distal area of flap of rats in two groups was similar, the inflammatory cells were infiltrated abundantly, and tissue edema was obvious. A large number of new blood vessels appeared in the middle area of flap of rats in bivalirudin group, with the formation of collateral vessels, and basic dilation of new blood vessels was seen. There were fewer new blood vessels appeared in the middle area of flap of rats in normal saline group, and dilation of new blood vessels was not obvious. There was little inflammatory cells infiltration in the proximal area of flap of rats in two groups. Compared with that in normal saline group, tissue edema extent of proximal area of flap of rats in bivalirudin group was less, and expansion was observed in more blood vessels. (4) The MVD of middle area of flap of rats in bivalirudin group was (26±5)/mm^2, significantly higher than that in normal saline group [(18±3)/mm^2, t=5.43, P<0.05]. (5) The expression of VEGF of middle area of flap of rats in bivalirudin group was 6 534±384, significantly higher than that in normal saline group (4 659±448, t=12.31, P<0.05). Conclusions Bivalirudin can promote the survival of random skin flap in rats, and the mechanisms may include reducing the formation of thrombosis, improving the blood supply of flap, and increasing the expression of VEGF, promoting the formation of new blood vessels.

The recombinant plasmid carrying the gene encoding 3C protease of Enterovirus 71 (EV71) was constructed, the recombinant protein was then expressed and purified, the functional activity was also measured. Firstly, the 3C protease gene was inserted into pET28a vector, the constructed recombinant plasmid was transformed into E. coli BL21 (DE3) for expression under the induction of IPTG. The expressed protein was purified by affinity chromatography (Ni-NTA) and the N-terminus His-tag was cleaved by enterokinase from 3C protease. The activity of 3C protease was evaluated with fluorescent peptide substrates. It was verified by restriction analysis and sequencing that recombinant plasmid pET28a-3C was constructed correctly and functionally expressed in E. coli BL21 (DE3) resulting in the production of recombinant 3C protease with a size of 22kD. Both His-tag and non-His-tag (cleaved by enterokinase) 3C protease exhibited similar enzyme activity to 3B-3C fluorescent peptide with Km, Vmax and Kcat values of 22 microM, 434nM. Min(-1) and 0.0669 Min(-1), respectively. The optimial pH and temperature were 7.0 and 30-37 degrees C, respectively. The acquirement of recombinant purified 3C protease with high activity has paved the way of further studies on anti-viral inhibitors, structural protein assembly, vaccine development and detection methods of EV71.
Cloning, Molecular ; Cysteine Endopeptidases ; chemistry ; genetics ; metabolism ; Enterovirus A, Human ; enzymology ; genetics ; Escherichia coli ; genetics ; metabolism ; Gene Expression ; Kinetics ; Viral Proteins ; chemistry ; genetics ; metabolism

OBJECTIVE:To study the quality standards of Compound kuqin ointment.METHODS:TLC method was used for the identification of alion,baicalin,menthol.The content of tetrahydropalmatine and baicalin were determined by HPLC.RESULTS:The identification method was specific.The linear range of tetrahydropalmatine was 0.131～3.275 ?g(r=0.999 9) with an average recovery of 102.40%(RSD=0.77%,n=6).The linear range of baicalin was 0.050 35～2.014 ?g (r=0.999 9)with an average recovery of 100.97%(RSD=1.42%,n=6).CONCLUSION:The method is simple,accurate and reproducible for the quality control of Compound kuqin ointment.