The antimicrobial peptide magainin II is expressed in the skin of the African clawed frog, Xenopus laevis, and exhibits a broad spectrum of antimicrobial activity as well as tumoricidal properties at low concentrations. In addition, magaininII plays a synergistic role during antimicrobial and tumoricidal processes with another antimicrobial peptide PGLa that is also expressed in Xenopus laevis. The optimized cDNA sequence of magainin II and magainin II-PGLa hybrid peptide according to E. coli or Pichia pastoris codon usage frequency were synthesized and sub-cloned into prokaryotic expression vector pGEX and Pichia pastoris secreted expression vector pPIC9k. The resulting recombinant plasmids were named as pGEX-magainin II and pPIC9k-magainin II-PGLa. The GST-magainin II fusion protein was highly expressed in E. coli. Furthermore, magainin II was successfully purified by digestion with PreScission Protease to cleave the GST tag. Additionally, our data obtained from the ELISA revealed that magainin II -PGLa hybrid peptide was successfully expressed in Pichia pastoris. These experiments establish a useful system for further studies of these antimicrobial peptides.
Animals ; Escherichia coli ; metabolism ; Genetic Vectors ; Magainins ; biosynthesis ; genetics ; Peptides ; genetics ; metabolism ; Pichia ; metabolism ; Plasmids ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Xenopus Proteins ; biosynthesis ; genetics ; Xenopus laevis

Objective:To explore the clinical effect of the second toenail flap with the dorsal bone of the phalange in repairing the defect of fingernail bed.Methods:From January, 2012 to June, 2019, 10 patients with large area of fingernail bed defect were treated by the second toenail flap with bone on the back of the phalanx. The survival of the flap was observed after the operation, and the fracture healing, the shape of the nail and the flexion and extension function of the finger joint were observed in the outpatient follow-up.Results:All flaps of the second toenail survived. The average follow-up period was 8 (4-12) months. The fractured ends of 10 patients' phalanges healed well without nonunion, good appearance of toenail and deformity of toenail. The recovery of hand function was evaluated according to the evaluation standard of upper limb function of Hand Surgery Society of Chinese Medical Association, 9 cases were excellent, and 1 case was good.Conclusion:The second toenail flap with dorsal bone of the phalanx preserved is easy to cut, simple to operate, and has good clinical effect. It is a good method to repair the defect of the fingernail bed.

Objective To explore the outcomes of free flap from the radial artery superficial palmar branch (RASP) innervated by the lateral cutaneous nerve of the forearm for repairing the defects of finger pulp. Methods From April, 2013 to February, 2015, 20 fingers in 20 cases were treated with free flap from RASP innervated by the lateral cutaneous nerve of the forearm. The area of flap was from 1.8 cm×2.0 cm to 2.0 cm×4.0 cm. Postoperative fol-low up was done termly. Results All 20 flaps survived. All cases were followed-up for 7-20 months ( averag, 12 months). The flaps appeared well with good texture. The two-point discrimination was from 7 mm to 12 mm, and grad-ed with S3 and S3+sensation. There was little scar noted at the donor site in the wrist. There was no influence of hand and wrist function. Conclusion The method of using free flap from SASP innervated by the lateral cutaneous nerve of the forearm to repair the defects of finger pulp has good clinical effect.

Objective:To investigate the clinical effect of Flow-through perforator flap of posterior interosseous artery in repair of dorsal digital soft tissue defect with disorder of blood supply in digital tip.Methods:From January 2018 to June 2020, 12 patients who had digital dorsum soft tissue defect with digital tip blood supply disorder were treated with Flow-through perforator flap of posterior interosseous artery. The size of flaps was 2.0 cm× 2.0 cm-5.5 cm×3.0 cm. The posterior interosseous artery in the flap was bridged with the proper palmar artery of digit, 1 subcutaneous vein in the flap was anastomosed with the dorsal subcutaneous vein, and 1 subcutaneous vein in the posterior interosseous artery with the palmar subcutaneous vein. The donor sites were sutured directly. The wound healing, blood supply of digit and flap survival were observed after operation. The quality of flap survival and digital joint function were observed in the follow-up reviews at outpatient clinic.Results:All the 12 Flow-through perforator flaps of posterior interosseous artery survived, the blood supply of digit was good, and the wounds healed in the first stage. The follow-up period was 6-24 months. The appearance and texture of the flaps were good without obvious bloating. Only linear scar was left in the forearm donor site. According to the Trial Standard of Upper Limb Function Evaluation of Chinese Society of Hand Surgery, the results were excellent in 11 cases and good in 1 case.Conclusion:The perforator flap of posterior interosseous artery has constant anatomy, and the diameter of blood vessel matches the blood vessels of digits. It is suitable for Flow-through technique. It has less subcutaneous tissue, no secondary thinning, and the donor site can be closed directly. It is a good method to repair the dorsal soft tissue defect with disorder of digital end blood circulation.

Objective:To investigate the clinical effect of microdissected peroneal artery perforator flap in repair of soft tissue defect of dorsal side of the fingers.Methods:From August 2015 to July 2020, 19 patients with soft tissue defects on dorsal fingers were treated with microdissected peroneal artery perforator flap. The area of wound defect was 3.8 cm×1.5 cm-5.8 cm×3.0 cm, with exposure of phalanges and tendons. The size of flaps was 4.0 cm×1.8 cm-6.0 cm×3.3 cm. According to the size of soft tissue defects on the dorsal side of the fingers, the flaps were designed with the perforating branch of peroneal artery in the centre. The length and width of a flap were 0.2-0.3 cm bigger and wider than the area of defect. The perforator vessels with a length of 2.0-3.0 cm were arvested in the superficial layer of deep fascia. Most of the adipose tissues of the flap were removed under microscope, and the small arteries between adipose tissues were protected. The flaps were used to cover the defects of fingers. The perforator artery of the flap was anastomosed with the proper palmar digital artery of the recipient site, the accompanying vein of the perforator artery was anastomosed with the dorsal digital vein of the recipient site, and the cutaneous nerve in the flap was anastomosed with the dorsal digital nerve. The donor sites were directly pulled together and sutured intermittently. Outpatient and WeChat follow-up were conducted after operation, including wound healing, flap survival, flap sensation, donor site recovery, and flexion and extension functions of the fingers. Functional recovery was evaluated according to the Evaluation Standard of Upper Limb Partial Functional of Hand Surgery of Chinese Medical Association.Results:All wounds healed in Ⅰ stage, and all 19 flaps survived. The follow-up ranged from 9 to 25 months, with an average of 11.5 months. The appearance of the flaps was satisfactory and the texture was good. Sensation recoveried to S 4 in 4 paitients, S 3 in 9 patients and S 2 in 6 patients, and with only a linear scar was left in the donor sites. The hand function recovery was evaluated according to the Trial Criteria of Upper Limb Function Evaluation of the Hand Surgery Society of the Chinese Medical Association, with 18 cases were excellent and 1 was good. Conclusion:The microdissected peroneal artery perforator flap is an ideal surgical method to repair the soft tissue defect of dorsal side of the fingers, which has good shape and simple operation, avoids the secondary thinning and plastic surgery and offers good therapeutic effects.

Objective:To investigate the surgical method and clinical effect in the repair of thumb tip defect with radial thenar fasciocutaneous flap.Methods:From March, 2016 to January, 2019, 15 patients with thumb tip defect were treated by radial thenar fasciocutaneous flap. There were 6 defects in left thumb and 9 in right thumb. All the thumbtip defects had the exposure of digital bone, and some had defects of digital bone itself. The size of defects ranged from 1.0 cm×2.0 cm to 1.5 cm×3.0 cm, and the size of the flaps ranged from 1.2 cm×2.5 cm to 1.8 cm×4.0 cm. All of the donor sites were directly closed. The patients entered follow-up by outpatient visit, telephone calls or WeChat reviews to evaluate the quality of the flaps and the clinical effects of the surgery.Results:All flaps survived with good quality. All the donor sites achieved primary healing. The follow-ups lasted for 6 to 18 months with an average of 10.5 months. At the final follow-up review, the appearance of flaps was satisfactory in natural colour, flexible and wear-resistant. There were various degrees of sensation recovery of the flaps, with TPD from 6 mm to 12 mm. The sensation around the donor site was close to normal. Function of the thumbs was evaluated according to the total active movement (TAM) evaluation system. Twelve thumbs were graded excellent and 3 in good.Conclusion:The radial thenar fasciocutaneous flap achieved satisfactory clinical effect in the repair of thumb tip defect. The texture of the flaps is close to the recipient site and the damage to the donor site is minimal.

Objective:To explore the clinical effect of a self-designed wound measurement grid on the design of lobulated perforator flap.Methods:From January 2019 to December 2022, soft tissue defects in limbs of 9 patients were reconstructed by lobulated perforator flaps in the Department of Orthopaedics and Traumatology of Shenzhen Hospital of Integrated Traditional Chinese and Western Medicine. The patients were 7 males and 2 females aged 21-55 years old with 40.3 years old in average. There were 6 upper limb defects and 3 lower limb defects. Self-designed wound measuring grids were applied to guide the design and excision of lobulated perforator flaps during surgery. Five patients were treated by lobulated perforator flap pedicled with the descending branch of lateral circumflex femoral artery and 4 by perforator flaps pedicled with dorsal interosseous artery. The areas of soft tissue defect were 4.0 cm×1.5 cm-26.0 cm×8.0 cm, and the sizes of the flaps were 4.5 cm×1.8 cm-22.0 cm×10.0 cm. After surgery, the blood supply of flaps, flap survival and wound healing were monitored. Flap survival, donor site recovery and limb function were observed at outpatient clinic over the postoperative follow-up, and the limb function was evaluated by the Evaluation Standard of Upper Limb Partial Functional of Hand Surgery of Chinese Medical Association and Lower Extremity Measure(LEM).Results:In this study, a total of 17 of 18 lobes of the lobulated perforator flaps survived after surgery. One flap had venous occlusion and after exploration and further treatment, a necrosis occurred in 1 lobe of the lobulated flap and then a free flap transfer was performed again. All the wounds achieved stage-I healing. Postoperative follow-up lasted for 3-18 months with 8.6 months in average. All flaps had good colour and texture, satisfactory appearance and only linear scars remained in the donor sites, without pain and scar contracture. The reconstructed upper extremity defect were evaluated according to the Evaluation Standard of Upper Limb Partial Functional of Hand Surgery of Chinese Medical Association, with 4 patients were in excellent and 2 in good. The reconstructed lower extremity defects were evaluated according to the LEM, and 2 patients were in excellent and 1 in good.Conclusion:The self-designed wound measurement grid was successfully applied in the design of lobed perforator flaps. It can accurately measure a wound surface and quickly extract a wound profile, and it is more convenient and intuitive to guide the design of flaps. It has a good effect in clinical application and further clinical trials are required.

The condition of patients with severe traumatic brain injury (sTBI) complicated by corona virus 2019 disease (COVID-19) is complex. sTBI can significantly increase the probability of COVID-19 developing into severe or critical stage, while COVID-19 can also increase the surgical risk of sTBI and the severity of postoperative lung lesions. There are many contradictions in the treatment process, which brings difficulties to the clinical treatment of such patients. Up to now, there are few clinical studies and therapeutic norms relevant to sTBI complicated by COVID-19. In order to standardize the clinical treatment of such patients, Critical Care Medicine Branch of China International Exchange and Promotive Association for Medical and Healthcare and Editorial Board of Chinese Journal of Trauma organized relevant experts to formulate the Chinese expert consensus on clinical treatment of adult patients with severe traumatic brain injury complicated by corona virus infection 2019 ( version 2023) based on the joint prevention and control mechanism scheme of the State Council and domestic and foreign literatures on sTBI and COVID-19 in the past 3 years of the international epidemic. Fifteen recommendations focused on emergency treatment, emergency surgery and comprehensive management were put forward to provide a guidance for the diagnosis and treatment of sTBI complicated by COVID-19.