[Clinical effects of antibiotic bone cement combined with free anterolateral thigh flap in sequential treatment of diabetic foot ulcer].

Objective: To investigate the clinical effects of antibiotic bone cement combined with free anterolateral thigh flap in sequential treatment of diabetic foot ulcer (DFU) wounds. Methods: A retrospective observational study was conducted. From August 2018 to August 2021, 15 patients with DFU who met the inclusion criteria were admitted to the Affiliated Hospital of Zunyi Medical University, including 12 males and 3 females, aged 42-65 years, with a history of type 2 diabetes for 5-19 years. All the wounds of patients were complicated with local bone, muscle, or tendon defects or exposure. The wounds were covered with antibiotic bone cement after debridement in stage Ⅰ+free anterolateral thigh chimeric perforator flap (perforator flap+muscle flap) or simple free anterolateral thigh flap grafting in stage Ⅱ. The defect area of the wound after bone cement removal and debridement was 9.0 cm×5.0 cm-20.0 cm×7.0 cm, the incision area of the flap was 10.0 cm×5.0 cm-22.0 cm×7.0 cm, and the incision area of the muscle flap was 5.0 cm×3.0 cm-8.0 cm×4.0 cm. The donor sites of flaps were sutured directly. During follow-up, the situations of donor site healing and flap survival were observed. At the last follow-up, the texture and shape of the flap, the presence of new ulcers on both limbs, and the walking ability of the patient were observed. Results: During the follow-up of 8 to 21 months after operation in stage Ⅱ, the donor sites healed well with only residual linear scar; flaps in 14 patients survived completely, and the flap in 1 patient developed partial necrosis at 3 weeks after stage Ⅱ surgery, which was healed after debridement and skin grafting. At the last follow-up, the flaps were good in texture and appearance, there were no new ulcers in the affected limb or opposite limb, and the patients had no obvious impairment in daily walking function. Conclusions: To repair DFU wounds with antibiotic bone cement combined with free anterolateral thigh flap can rapidly control the infection, achieving a high survival rate of flap after operation with no obvious impairment in daily walking function of patients.

[Research advances on the mechanism of nerve regeneration-related protein in skin fibrosis].

The healing process after skin injury is a dynamic process of interaction between various cells, cytokines, and extracellular matrix. Fibrosis is one of the main ways of skin injury repair. The process of fibrosis involves the regulation of many factors. Studies have shown that nerve regeneration-related protein (NREP) plays a key role in the fibrosis of skin tissue and organs. Based on the mechanism of skin fibrosis, this paper discusses the construction of tertiary structure of NREP, summarizes the effects of NREP and different cells in the skin on skin fibrosis and the research progress of mechanism of NREP in skin fibrosis, thus providing new ideas for the treatment of skin fibrosis diseases.

[Clinical effects of superficial temporal artery lobulated perforator flaps in repairing skin and soft tissue defects after temporal tumor resection].

Objective: To explore the feasibility and clinical effects of using superficial temporal artery lobulated perforator flaps in repairing skin and soft tissue defects after tumor resection in the temporal region. Methods: A retrospective observational study method was used. From March 2017 to October 2022, ten patients with temporal skin tumors were admitted to the Affiliated Hospital of Zunyi Medical University, including six women and four men, with age ranging from 42 to 87 years. Among them, three patients had squamous cell carcinoma and seven patients had basal cell carcinoma, with disease duration ranging from 6 months to 5 years. All temporal tumors underwent expanded resection, leaving wound areas of 5.4 cm×4.2 cm to 7.0 cm×4.0 cm after tumor resection. Superficial temporal artery frontal branch flaps with areas of 5.5 cm×1.2 cm to 7.0 cm×1.5 cm, superficial temporal artery descending branch flaps with areas of 4.2 cm×3.5 cm to 5.0 cm×4.0 cm, and superficial temporal artery parietal branch flaps with areas of 4.2 cm×1.0 cm to 5.0 cm×1.0 cm were designed to repair the wounds and reconstruct the hairline. The donor areas of the flaps were closed and sutured directly. The survival of the flaps was observed on 3 to 5 days after surgery, and the healing of wounds on the donor and recipient sites was observed when the stitches were removed on 5 to 7 days after surgery. During follow-up after surgery, the appearance of the temporal area, scar hyperplasia, hairline reconstruction, and tumor recurrence were observed in the temporal region on the affected side. Results: All the flaps survived well on 3 to 5 days after surgery, and all the donor and recipient site wounds healed well on 5 to 7 days after surgery. During follow-up of 3 to 6 months after surgery, the surgical incisions were concealed; the flaps were not swollen, with a consistent color to the surrounding skin; there were no obvious hypertrophic scars; the reconstructed hairline on the affected side was not significantly different from that of the healthy side; there was no tumor recurrence in the local area. Conclusions: For large areas of skin and soft tissue defects in the temporal region, the use of superficial temporal artery lobulated perforator flaps can repair the wounds in different regions and suture the donor sites in the primary stage simultaneously. The surgical operation is simple, and the facial appearance conforms to the aesthetic requirement after surgery with no tumor recurrence in the local area but a good repair effect. This method is particularly suitable for repairing large areas of skin and soft tissue defects in the temporal region in elderly patients.

[Clinical application effects of two longitudes three transverses method in perforator location of thoracodorsal artery perforator flap and deep wound repair].

Objective: To explore the clinical application value of two longitudes three transverses method in the location of the perforator of thoracodorsal artery perforator and deep wound repair. Methods: The retrospectively observational study was conducted. From December 2018 to June 2020, 17 patients with deep wounds who were admitted to the Affiliated Hospital of Zunyi Medical University met the inclusion criteria and were included in this study, including 7 males and 10 females, aged 12 to 72 years. The wound areas of patients after debridement were 7 cm×3 cm to 11 cm×7 cm. Two longitudinal lines were located through the midpoint of the armpit, the posterior superior iliac spine, and the protruding point of the sacroiliac joint, and three transverse lines were located 5, 10, and 15 cm below the midpoint of the armpit between the two longitudinal lines, i.e. two longitudes three transverses method, resulting in two trapezoidal areas. And then the thoracodorsal artery perforators in two trapezoidal areas were explored by the portable Doppler blood flow detector. On this account, a single or lobulated free thoracodorsal artery perforator flap or flap that carrying partial latissimus dorsi muscle, with an area of 7 cm×4 cm to 12 cm×8 cm was designed and harvested to repair the wound. The donor sites were all closed by suturing directly. The number and location of thoracodorsal artery perforators, and the distance from the position where the first perforator (the perforator closest to the axillary apex) exits the muscle to the lateral border of the latissimus dorsi in preoperative localization and intraoperative exploration, the diameter of thoracodorsal artery perforator measured during operation, and the flap types were recorded. The survivals of flaps and appearances of donor sites were followed up. Results: The number and location of thoracodorsal artery perforators located before operation in each patient were consistent with the results of intraoperative exploration. A total of 42 perforators were found in two trapezoidal areas, with 2 or 3 perforators each patient. The perforators were all located in two trapezoid areas, and a stable perforator (the first perforator) was located and detected in the first trapezoidal area. There were averagely 1.47 perforators in the second trapezoidal area. The position where the first perforator exits the muscle was 2.1-3.1 cm away from the lateral border of the latissimus dorsi. The diameters of thoracodorsal artery perforators were 0.4-0.6 mm. In this group, 12 cases were repaired with single thoracodorsal artery perforator flap, 3 cases with lobulated thoracodorsal artery perforator flap, and 2 cases with thoracodorsal artery perforator flap carrying partial latissimus dorsi muscle. The patients were followed up for 6 to 16 months. All the 17 flaps survived with good elasticity, blood circulation, and soft texture. Only linear scar was left in the donor area. Conclusions: The two longitudes three transverses method is helpful to locate the perforator of thoracodorsal artery perforator flap. The method is simple and reliable. The thoracodorsal artery perforator flap designed and harvested based on this method has good clinical effects in repairing deep wound, with minimal donor site damage.

[Effects of free mini-flap on tibial side of third toe on repairing skin and soft tissue defect of finger pulp at the end of finger].

Objective: To investigate the effects of free mini-flap on tibial side of third toe on repairing skin and soft tissue defect of finger pulp at the end of finger. Methods: From August 2013 to May 2017, 18 patients with skin and soft tissue defect of finger pulp at the end of finger were admitted to our unit, with 12 men and 6 women aged 16 to 54 years. As the skin and soft tissue defect sites, there were 3 cases of thumb, 8 cases of index finger, 4 cases of middle finger, and 3 cases of ring finger. The area of defects ranged from 2.0 cm×1.4 cm to 3.5 cm×2.4 cm. Free mini-flaps on tibial side of third toes were designed according to area and shape of defects, and the length and width of flaps were 0.1 to 0.2 cm longer than the length and width of the defects, respectively. The area of flaps ranged from 2.1 cm×1.5 cm to 3.7 cm×2.6 cm. The end-to-end anastomosis of subcutaneous veins of flaps and superficial veins of the finger-palm side or superficial dorsal digital vein, the end-to-end tension-free anastomosis of the base metatarsal arteries on tibial side of third toe and proper digital arteries of recipient finger were performed. Besides, anastomosis of base metatarsal nerve on tibial side of third toe and proper digital nerve of recipient finger was performed. The donor sites on feet were sutured directly or repaired with full-thickness skin grafts on medial upper leg of the same side. The survival of flaps after operation and the follow-up of patients were observed. Results: All flaps survived well, with good blood supply. Among the 18 patients, 2 patients lost to follow-up, and 16 patients were followed up for 4 to 36 months. The shape and texture of flaps were good. After reconstruction, finger pulps at the end of finger were plump, with fingerprint. Function of the finger restored well, and the two-point discriminatory distances of flaps were 5 to 10 mm. The donor sites on feet of 14 patients healed after the operation, the other 2 patients had necrosis on edge and central area of skin grafts, and the necrotic area healed after dressing change. The skin graft areas on feet were wear-resistant, with slight damage to donor sites and did not influence shoes wearing and walking. Besides, patients did not feel uncomfortable. Conclusions: Skin and soft tissue defects of finger pulp at the end of finger repaired by free mini-flaps on tibial side of third toe are with good shape and slight damage to donor sites, and the operation is simple. It is worthy of popularization and application in clinic.

[Effects of combined transplantation of rat Schwann cells and fibroblasts on nerve regeneration of denervated perforator flaps in rats and the mechanism].

Objective: To explore the effects of combined transplantation of the rat Schwann cells and fibroblasts (Fbs) on the nerve regeneration of denervated perforator flaps in rats and the mechanism. Methods: (1) Fbs were isolated from the trunk of 2 Sprague-Dawley (SD) rats embryos of 14-16 days' pregnancy and cultured, and the morphology of the cells was observed. The third passage of cells were used for subsequent experiments. The protein expressions of fibronectin and Ephrin-B2 were observed by immunohistochemical method. The mRNA expression of Ephrin-B2 was detected by real-time fluorescent quantitative reverse transcription polymerase chain reaction (n=3). (2) Schwann cells were isolated from the bilateral sciatic nerves and brachial plexus nerves of 45 SD rats born for 1-3 days and cultured, and the morphology of the cells was observed. The third passage of cells were used for subsequent experiments. The rate of S100 positive cells was detected by immunofluorescence method and flow cytometer, with sample numbers of 9 and 3 respectively. (3) In Dulbecco's modified Eagle medium (DMEM) high glucose medium, 1 mL Fbs and 1 mL Schwann cells both in the concentration of 1×10(5) cells/mL were co-cultured as Schwann cells+ Fbs co-culture group, and 2 mL Schwann cells in the concentration of 1×10(5) cells/mL were cultured alone as Schwann cells alone culture group, with 5 wells in each group. The clusters of Schwann cells in the two groups were observed and counted under inverted phase contrast microscope at post culture hour (PCH) 6 and 24 respectively. The clusters of Schwann cells in Schwann cells+ Fbs co-culture group were observed by immunofluorescence method at PCH 24 too. The protein expressions of EphB2, Sox2, and N-cadherin in Schwann cells of two groups at PCH 24 were detected by Western blotting (n=20). (4) Totally 100 8-week-old male SD rats were selected, and an in situ replanted peritoneal denervated perforator flap was made in each rat. According to the random number table, the rats were divided into simple flap group, Fbs alone transplantation group, Schwann cells alone transplantation group, Schwann cells+ Fbs co-transplantation group, with 25 rats in each group. Flaps of rats in Fbs alone transplantation group and Schwann cells alone transplantation group were injected with 0.4 mL Fb and 0.4 mL Schwann cells respectively (2×10(6) cells each). Flaps of rats in Schwann cells+ Fbs co-transplantation group were injected with 0.4 mL Fbs and Schwann cells mixed cells (totally 2×10(6) cells, cell number ratio: 1∶1), and flaps of rats of simple flap group were injected with the same volume of DMEM high glucose medium. On post injection day (PID) 2, 5, 7, 9, and 14, 5 rats in each group were selected respectively according to the random number table. The flap tissue was collected, and the number, diameter, and arrangement of regenerated nerves were observed by immunofluorescence method. Data were processed with completely random designed t test, analysis of variance for repeated measurement, t test, and Bonferroni correction. Results: (1) The third passage of cells isolated and cultured from the rat embryo trunks were uniform in size and shape, long spindle-shaped, with a large proportion of nuclei. Strong positive expressions of fibronectin and Ephrin-B2 protein in cells were observed, and the mRNA expression of Ephrin-B2 was 0.004 1±0.000 8. The cells were identified as Fbs. (2) After 5 days of culture, the primary cells isolated from the sciatic nerves and brachial plexus nerves of neonatal rats were elongated in cell bodies and grew in nest, fence, or vortex-like shape. The third passage of cells were detected by immunofluorescence method and flow cytometer, and the corresponding S100 positive cell rates were (95.9±1.0)% and (95.8±1.1)% respectively. The cells were identified as Schwann cells. (3) At PCH 6 and 24, the cluster numbers of Schwann cells in Schwann cells+ Fbs co-culture group were significantly higher than those of Schwann cells alone culture group (t=6.500, 10.614, P<0.01). At PCH 24, the Schwann cells in Schwann cells+ Fbs co-culture group aggregated into clusters, Fbs dispersed around the Schwann cell clusters, and the protein expressions of EphB2, N-cadherin, and Sox2 in Schwann cells were significantly higher than those in Schwann cells alone culture group (t=2.975, 19.717, 11.159, P<0.05 or P<0.01). (4) On PID 2, a small number of scattered, disordered, short, and thin nerve fibers were observed in the flap tissue of rats in the four groups. From PID 5 to 14, the number of nerve fibers in the flap tissue of rats of Schwann cells+ Fbs co-transplantation group increased gradually, and the nerve fibers were with long diameter and arranged orderly. The number of nerve fibers in the flap tissue of rats of Schwann cells alone transplantation group increased, but the nerve fibers were with short diameter and arranged disorderly, and the number was smaller than that of Schwann cells+ Fbs co-transplantation group. In simple flap group and Fbs alone transplantation group, the nerve fibers in the flap tissue of rats gradually degenerated with gradually decreased number or even disappeared. Conclusions: The combined transplantation of Fbs and Schwann cells in rats can regulate Schwann cells migration and clustering by activating Ephrin/Eph-Sox2-N-cadherin signaling pathway, thus promoting the orderly nerve regeneration of denervated perforator flaps in rats.

[Clinical effects of middle and low peroneal artery perforator flap with pedicle on repairing skin and soft tissue defects of ankle].

Objective: To investigate clinical effects of middle and low peroneal artery perforator flap with pedicle on repairing skin and soft tissue defects of ankle. Methods: Twenty patients with skin and soft tissue defects of ankle and exposure of tendon and bone were admitted in our burn wards from April 2012 to December 2015. The size of skin and soft tissue defects ranged from 5 cm×4 cm to 23 cm×10 cm. Patients were treated with debridement and vacuum sealing drainage (VSD) after admission. After VSD treatment for 1 week, flap transplantation operation was performed. Middle and low perforating branches of peroneal artery were detected by portable Doppler blood flow meter before the operation. Flaps were designed and resected according wounds during the operation, with 1 or 2 middle and low perforating branches of peroneal artery in flaps. Seventeen patients were treated with middle and low peroneal artery perforator flap. Larger wounds with exposure of tendon and bone were repaired with middle and low peroneal artery perforator flap, and the other wounds were repaired with intermediate split-thickness skin graft of thigh on the same side in three patients. The size of flap ranged from 6 cm×5 cm to 25 cm×12 cm. The donor sites were sutured directly or repaired with intermediate split-thickness skin graft of thigh on the same side. Results: After operation, 1 patient with partial skin necrosis at the distal of the flap because of disorder of venous circulation healed after dressing change and physiotherapy, and flaps of the other 19 patients survived well. During follow-up of 3 to 36 months, flaps of all patients were in good appearance, with no obvious cicatrix, and the affected limbs and ankle joints functioned well. Conclusions: Middle and low peroneal artery perforator flap with advantages of stable perforating branch, reliable blood supply, and large resected size, can repair skin and soft tissue defects of ankle.