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Osteoarthritis is a common degenerative joint disease, and cartilage damage is often considered an early factor in irreversible joint degeneration. Repairing damaged cartilage remains a medical challenge due to its limited ability to self-repair and regenerate. In recent years, the application of tissue engineering strategies to treat cartilage defects has been recognized as an emerging therapeutic avenue. Acellular cartilage matrix (ACM) is an ideal material for cartilage repair and regeneration as it retains the extracellular matrix structure and bioactive components of natural cartilage, mimicking the extracellular environment of natural cartilage to the greatest extent. Type II collagen is the main type of hyaline cartilage and plays an important role in regulating the mechanical properties of cartilage tissue. It has been shown that type II collagen, growth factors and the hypoxic microenvironment play important roles in promoting cartilage regeneration. Type II collagen induces cell aggregation and chondrogenic differentiation in a specific way; Various growth factors contained in the ACM induce Sox9 expression and promote chondrogenic differentiation of stem cells; The hypoxic microenvironment upregulates the expression of type II collagen (COL2A1), Sox9 and maintains chondrocyte phenotype. In addition, ACM has been widely used in cartilage regeneration studies, either as a decellularized scaffold, hydrogel or 3D bioprinting technique for the repair of defective cartilage. Although the ACM-derived biomaterials discussed in this paper have many advantages, there are still some difficulties in their practical applications, such as loss of ACM components and reduced scaffold performance, which are still worth exploring in depth.
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Objective:To analyze the necessity of anastomosis of the cutaneous nerve by comparing anterolateral femoral flaps with versus without anastomosis of the anterolateral femoral cutaneous nerve in the repair of extremity soft tissue defects.Methods:A retrospective analysis was made of the clinical data of 30 patients with extremity soft tissue defects who had been admitted to Department of Orthopedics, The Sixth People＇s Hospital affiliated to Shanghai Jiaotong University School of Medicine from September 2019 to March 2022. The patients were assigned into 2 groups according to whether the anterolateral femoral cutaneous nerve was anastomosed or not in the repair of extremity soft tissue defects using anterolateral femoral flaps. In the anastomosis groups of 14 cases, there were 7 males and 7 females, with an age of (46.4±15.2) years and a flap size of (22.4±7.3) cm×(8.5±1.3) cm. In the non-anastomosis group of 16 cases, there were 11 males and 5 females, with an age of (39.9±15.8) years and a flap size of (23.0±6.4) cm×(9.0±2.1) cm. The 2 groups were compared in terms of flap survival, Semmes-Weinstein recovery degree and area of monofilament tactile sensation, and time periods for temperature sensation and two-point discrimination.Results:There was no statistically significant difference between the 2 groups in the preoperative general data, showing they were comparable ( P>0.05). All the flaps survived completely without vascular crisis. In the anastomosis group, the time periods required for Semmes-Weinstein recovery of monofilament tactile sensation to the areas of 20.0%, 50.0%, and 80.0% [(2.5±0.7) months, (6.7±1.1) months, and (11.0±1.2) months] were significantly shorter than those in the non-anastomosis group [(3.6±1.3) months, (8.6±1.4) months, and (15.0±2.2) months], the recovery area at the last follow-up [100.0% (100.0%, 100.0%)] was significantly larger than that in the non-anastomosis group [84.6% (81.7%, 89.9%)], and the time period for recovery of temperature sensation [(3.9±0.7) months] significantly shorter than that in the non-anastomosis group [(6.1±1.1) months] (all P<0.05). The time for recovery of two-point discrimination in the 14 patients in the anastomosis group was (10.4±1.7) months while only 7 of the 16 patients in the non-anastomosis group recovered two-point discrimination after (14.7±1.4) months, showing a significant difference between the 2 groups ( P<0.05). Conclusion:In the repair of extremity soft tissue defects using anterolateral femoral flaps, compared with no anastomosis of the cutaneous nerve, anastomosis of the anterolateral femoral cutaneous nerve may ensure more or less the sensory recovery of the flaps.
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Objective To explore the operative technique and clinical results of posterior tibial artery perforator flap within saphenous nerve branch for sensory reconstruction.Methods From January,2016 to June,2018,9 patients suffered from soft tissue defect were treated by the posterior tibial artery perforator flap containing saphenous nerve branch.Seven patients were males and 2 were females,with age ranged from 31 to 62 years.Soft tissue defects located in hands in 5 patients,plantar in 2 patients,ankle in 1 patient and dorsal foot in 1 patient.The size of soft tissue defects ranged from 8.0 cm×2.5 cm to 21.0 cm×4.0 cm.The regular post-operative followed-up was performed.Results All flaps survived without complications.The size of flap ranged from 10.0 cm×3.5 cm-23.0 cm×5.0 cm.Donor sites were primarily closed in 5 patients and secondary closed in 4 patients.Followed-up ranged from 6 to 15 months with 10 months in average.The contour of flaps were satisfied and the sensory function of the donor sites were normal.At 6 months followed-up,SW test reached 5.07 in all flaps,and 2PD ranged from 14 to 35 mm.Conclusion The novel sensory flap can provide satisfied sensory outcome without sacrificing main artery and saphenous nerve,and is a good candidate for sensory reconstruction of soft tissue defects.
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Objective To investigate the feasibility and clinical effect of palmar venous anastomosis in fingertip replantation.Methods From October,2008 to May,2017,the clinical data of 15 patients with 24 finger replantation were retrospectively analyzed by outpatient,telephone and WeChat followed-up.One artery was anastomosed for revascularization with or without nerve repair,and 1 palmar venous anastomosis was performed to reestablish the outflow system.The finger nerve was sutured randomly according to the injury.Results The patients were followed-up for 18-65 (mean,39.9) months.Except for 1 replantation finger with rotation avulsed injury failed,the remaining 14 cases with 23 fingers survived completely.Excellent restoration of finger motion was observed during the follow-up.The mean regained static 2-point discrimination sensation was 3.6 (2.8-4.0) mm.All patients were satisfied with the treatment effect.Conclusion Successful palmar venous anastomosis can improve the survival rate of replanted fingertips,simplify postoperative care,reduce postoperative complications,and improve the treatment effect.
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Objective@#To evaluate osteotomy, bone shortening and lengthening by one stage with external fixation for a large bone defect following open femoral fracture.@*Methods@#From April 2014 to April 2019, 11 consecutive patients with a large bone defect following open femoral fracture were treated at Department of Orthopaedics, The Sixth People’s Hospital of Shanghai. They were 9 males and 2 females, with an average age of 40.5 years (from 18 to 70 years). The left side was involved in 6 cases and the right side in 5. All their primary open fractures had deteriorated into osteomyelitis and bone defects after treatment. Their bone defects averaged 60.5 mm after thorough debridement. Postoperative X-ray examinations were taken regularly. Bony union time and index, time till removal of external fixator, external fixation index, flap survival, soft tissue healing, visual analogue scale (VAS), Association for Studying and Application of Methods of Ilizarov (ASAMI) scores for bone healing and lower limb function were recorded.@*Results@#All the patients were followed up for 9 to 30 months (mean, 21.9 months). The bony union time averaged 10.5 months (from 5.2 to 22.3 months). The bony union indexes averaged 52.9 d/cm (from 33.4 to 73.3 d/cm). The time till removal of external fixator averaged 15.5 months (from 9.8 to 27.5 months). The external fixation indexes averaged 86.0 d/cm (from 60 to 113 d/cm). All the bone nonunions healed with no recurrence of osteomyelitis. Complications like union failure, refracture at ends of bone lengthening or fracture, or leg length discrepancy>2.5 cm, happened in none. The VAS scores ranged from 0 to 3 points. By ASAMI evaluation, the bony union was excellent in 10 cases and good in one case, and the lower limb function was excellent in 3 cases, good in 7 cases and fair in one.@*Conclusion@#Osteotomy, bone shortening and lengthening by one stage with external fixation is a reliable treatment for large femoral bone defects because this strategy can remove the lesions and restore the femoral length at the same time.
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Objective@#To report our clinical application of free fibular composite tissue flap with peroneus longus and brevis in the treatment of complicated extremity trauma.@*Methods@#From August 2014 to September 2017, 5 patients with complicated extremity trauma were treated using a free fibular composite tissue flap with peroneus longus and brevis at Department of Orthopaedics, The Sixth People's Hospital of Shanghai. They were 4 men and one woman, aged from 23 to 52 years (average, 35.1 years). All of them had tendon defects; one had a radius defect, one an ulnar defect and three a tibial defect. The length of bone defects ranged from 8 cm to 18 cm; the size of soft tissue defects ranged from 10 cm × 5 cm to 18 cm ×8 cm. A fibular composite tissue flap was designed according to the soft tissue condition and defect area to repair and fixate the bone defect after thorough debridement and cover the wound before functional reconstruction. Postoperatively, survival of the flap and functional recovery of the corresponding tendon at the recipient site, healing time of the tibia, and wound healing, ankle motion and complications at the donor site were all observed.@*Results@#The 5 patients were followed up for 18 to 38 months (average, 25.2 months). All flaps survived without any vascular crisis or infection. The union time for the fibular graft ranged from 4 to 16 months (average, 8.4 months). In the 3 patients with a tibial defect, the maximum angle of dorsal extension ranged from 0° to 10° and no foot drop was observed. Thumb and digital flexion was reconstructed in the patients with a forearm bone defect only to achieve opposition of index finger and thumb but we failed to make them have a fist. At the 5 flap donor sites, the wound was sutured by the second stage after vacuum suction and healed well; obvious scar formed in one of them. No obvious foot varus was observed at the donor site. The patients were satisfactory with their ankle joint motion. No refracture of the fibular graft occurred during follow-up.@*Conclusion@#A free fibular composite tissue flap with peroneus longus and brevis is a good choice for complicated extremity trauma.
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Objective To evaluate osteotomy,bone shortening and lengthening by one stage with external fixation for a large bone defect following open femoral fracture.Methods From April 2014 to April 2019,11 consecutive patients with a large bone defect following open femoral fracture were treated at Department of Orthopaedics,The Sixth People's Hospital of Shanghai.They were 9 males and 2 females,with an average age of 40.5 years (from 18 to 70 years).The left side was involved in 6 cases and the right side in 5.All their primary open fractures had deteriorated into osteomyelitis and bone defects after treatment.Their bone defects averaged 60.5 mm after thorough debridement.Postoperative X-ray examinations were taken regularly.Bony union time and index,time till removal of external fixator,external fixation index,flap survival,soft tissue healing,visual analogue scale (VAS),Association for Studying and Application of Methods of Ilizarov (ASAMI) scores for bone healing and lower limb function were recorded.Results All the patients were followed up for 9 to 30 months (mean,21.9 months).The bony union time averaged 10.5 months (from 5.2 to 22.3 months).The bony union indexes averaged 52.9 d/cm (from 33.4 to 73.3 d/cm).The time till removal of external fixator averaged 15.5 months (from 9.8 to 27.5 months).The external fixation indexes averaged 86.0 d/cm (from 60 to 113 d/cm).All the bone nonunions healed with no recurrence of osteomyelitis.Complications like union failure,refracture at ends of bone lengthening or fracture,or leg length discrepancy ＞ 2.5 cm,happened in none.The VAS scores ranged from 0 to 3 points.By ASAMI evaluation,the bony union was excellent in 10 cases and good in one case,and the lower limb function was excellent in 3 cases,good in 7 cases and fair in one.Conclusion Osteotomy,bone shortening and lengthening by one stage with external fixation is a reliable treatment for large femoral bone defects because this strategy can remove the lesions and restore the femoral length at the same time.
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Objective To report our clinical application of free fibular composite tissue flap with peroneus longus and brevis in the treatment of complicated extremity trauma.Methods From August 2014 to September 2017,5 patients with complicated extremity trauma were treated using a free fibular composite tissue flap with peroneus longus and brevis at Department of Orthopaedics,The Sixth People's Hospital of Shanghai.They were 4 men and one woman,aged from 23 to 52 years (average,35.1 years).All of them had tendon defects;one had a radius defect,one an ulnar defect and three a tibial defect.The length of bone defects ranged from 8 cm to 18 cm;the size of soft tissue defects ranged from 10 cm × 5 cm to 18 cm × 8 cm.A fibular composite tissue flap was designed according to the soft tissue condition and defect area to repair and fixate the bone defect after thorough debridement and cover the wound before functional reconstruction.Postoperatively,survival of the flap and functional recovery of the corresponding tendon at the recipient site,healing time of the tibia,and wound healing,ankle motion and complications at the donor site were all observed.Results The 5 patients were followed up for 18 to 38 months (average,25.2 months).All flaps survived without any vascular crisis or infection.The union time for the fibular graft ranged from 4 to 16 months (average,8.4 months).In the 3 patients with a tibial defect,the maximum angle of dorsal extension ranged from 0° to 10° and no foot drop was observed.Thumb and digital flexion was reconstructed in the patients with a forearm bone defect only to achieve opposition of index finger and thumb but we failed to make them have a fist.At the 5 flap donor sites,the wound was sutured by the second stage after vacuum suction and healed well;obvious scar formed in one of them.No obvious foot varus was observed at the donor site.The patients were satisfactory with their ankle joint motion.No refracture of the fibular graft occurred during follow-up.Conclusion A free fibular composite tissue flap with peroneus longus and brevis is a good choice for complicated extremity trauma.
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Objective To evaluate the outcomes of internal fixation combined with bone grafting in the treatment of distal humeral nonunion.Methods This retrospective study included 48 patients who had undergone internal fixation combined with iliac bone grafting for distal humeral nonunion between January 2011 and December 2015 at Department of Orthopedic Surgery,The Sixth People's Hospital of Shanghai.They were 31 males and 17 females,with a mean age of 35.4 years (from 22 to 49 years).The outcomes were evaluated by clinical examination,X-ray film,Mayo elbow performance score (MEPS) and visual analogue scale (VAS) during follow-up.Results All the 48 patients were followed up for 18 to 60 months (mean,36 months).No wound-related complications were reported in this cohort.Solid bone union was achieved at 3 to 8 months (mean,4.2 months) after surgery.At the last follow-up,the range of elbow flexion-extension was improved from preoperative 66° to 101°,and the ranges of pronation and supination were enhanced from 65° and 45° preoperatively to 82° and 75°,respectively.The MEPS increased from 54 points preoperatively to 82 points postoperatively.The results were excellent in 26,good in 16 and fair in 6 cases (with an excellent to good rate of 87.5％).The VAS decreased from 4.5 points preoperatively to 1.2 points postoperatively,indicating notable relief of the elbow pain due to distal humeral nonunion.Follow-ups revealed no ulnar nerve injury,nonunion,heterotopic ossification,implant loosening or breakage,or instability of the elbow joint.Conclusion Internal fixation combined with bone grafting is reliable in treatment of distal humeral nonunion,leading to satisfactory outcomes.
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Objective To report clinical application of free chimeric vascularized fibular graft combined with sural flap for reconstruction of composite extremity defects after open fracture.Methods From June 2010 to July 2014,free chimeric vascularized fibular grafts and sural flaps were used to treat 4 patients with composite extremity defects at Department of Orthopaedics,The Sixth People's Hospital.They were 3 men and one woman,aged from 39 to 61 years(average,48.5 years).There were 2 cases of soft tissue defects on the forearm complicated with radial defect,one case of soft tissue defects on the forearm complicated with ulnar shaft defect,and one case of soft tissue defects on the leg complicated with tibial defect.The length of bone defect ranged from 8 cm to 18 cm (average,13.0 cm);the size of soft tissue defects ranged from 22 cm × 6cmto23cm × 15 cm (average,22.3cm × 9.7cm).Results The area of flap ranged from 25 cm × 9 cm to 26 cm × 18 cm (average,25 cm × 13 cm);the length of fibular graft ranged from 8 cm to 18 cm (average,13 cm).The 4 patients were followed up for 8 to 42 months (average,20.5 months).All the chimeric flaps survived.All the fractures united after an average of 8.5 months.The last follow-ups revealed no refracture.All the patients were satisfied with the outcomes.Conclusion Free chimeric vascularized fibular graft combined with sural flap is a reliable choice for reconstruction of composite extremity defects after open fracture.
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Objective To determine the outcome of the combined use of flaps transfer and ilizarov technique reconstruct the large soft tissue defects and bone lose in the lower leg.Methods Sixteen patients were identified from a retrospective review from July,2008 to July,2013,who suffered the large soft tissue defects and bone lose in the lower leg and underwent single-stage soft tissue and osseous reconstruction using the flap technique and Ilizarov method.There were 12 males and 4 females aged from 22 to 62 years old (average 42.6 years old).The size of soft tissue defect ranged from 8 cm×9 cm to 30 cm×20 cm.The length of the bone discrepancy ranged from 2 to 14 cm.According to the local condition of the lower leg and the size of the composite tissue defects,10 patients received the free flap covering,6 patients repaired by the saphenous neurocutaneous perforator flap (3 cases) and sural nerve neurocutaneous flap (3 cases).Daily monitoring the skin temperature postoperative.Distraction was commenced on postoperative day 10 to 14 at the rate of 1 mm/day and continued in 4 equal increments.Results The follow-up time ranged from 18 to 36 months.Sixteen flaps survived completed,only 1 flap was observed the venous congestion in postoperative day 2.The duration of ilizarov application ranged from 3.5 to 18.0 months.All patients achieved final union.All patients were satisfied with the outcome of the surgery.Conclusion The combined use of neurocutaneous flap and Ilizarov technique for reconstruction of large composite soft tissue defect in the lower leg.Significantly reduce patient treatment time,improving traction osteogenesis of long bones and the ability of resistance to infection.
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BACKGROUND:Studies have shown that the vacuum sealing drainage technology can effectively promote the wound healing, and it has a wide prospect of clinical application, but there are few reports addressing the treatment of diabetic foot. OBJECTIVE:To discuss the clinical effect of vacuum sealing drainage technology in the treatment of diabetic foot wounds. METHODS: Sixty diabetic foot patients were randomly divided into two groups: traditional treatment group, regulating blood sugar level, dressing and traditional debridement; vacuum sealing drainage group, conventional treatment combined with the vacuum sealing drainage technology. The clinical efficacy of two treatments for diabetic foot was evaluated. RESULTS AND CONCLUSION: Compared with the traditional treatment group, the vacuum sealing drainage showed better outcomes in switching frequency, stable blood sugar control, preparation time, wound healing time and cure rate (P < 0.05). It indicates that the vacuum sealing drainage technology in the treatment of diabetic foot ulcers can resolute wound inflammation, stimulate the growth of granulation, create a favorable surgical condition for secondary skin grafting or flap coverage, significantly shorten the treatment time, and exhibit better curative effects than the traditional treatment.
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Objective To evaluate the clinical effect of the reversed saphenous fasciocutaneous island flap combined with the VSD therapy in plate exposure of the distal tibia.Methods From January 2008 to July 2010,seven patients with hardware exposure following internal fixation of the distal tibia fracture were treated by VSD therapy and reversed saphenous fasciocutaneous island flap reconstruction.The VSD therapy was performed from 7 to 26 days after hardware exposure,and the flap transfer was performed 7 to 14 days after the VSD therapy.The size of the soft-tissue defect ranged from 4 cm × 2 cm to 13 cm × 4 cm.The mean follow-up was 10.6 months after the fracture (range,8-14 months).Results The average times of the VSD therapy was 1.3.After VSD was removed,the exposed hardware was covered by healthy granulation tissue in all cases.All flaps were successfully transplanted,and the size of flap ranged from 6 cm × 3 cm to 15 cm × 6 cm.Six of 7 flaps survived completely without further procedures.The necrosis of the distal margin of the flap occurred in 1 patient.A solid bony union was confirmed by the X-ray in all patients after 4-6 months postoperatively.No skin defects or fistulae were observed.Conclusion The combination of the reversed saphenous fasciocutaneous island flap and VSD therapy could save the exposed hardware and cover the soft tissue defects in the distal tibia.
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Objective To study the anatomy of superficial palmar digital veins in fingertip,and explore the clinical application of superficial palmar digital veins as venous drainage in fingertip replantation.Methods One hundred fingers were studied with three methods: microanatomy,sectional anatomy,and X-ray.According to microanatomical observation,thirteen fingers in 10 cases with fingertip amputations and dorsal veins defect were replanted by anastomosis of palmar digital veins to reconstruct the venous drainage of the amputated digits. Results ( 1 ) 1 to 3 tiny superficial palmar digital veins can be found at the level of the onychorrhiza.The average diameters of the superficial palmar digital veins were 0.2-0.5 mm.(2)In clinical practice,13 replanted fingers were survived.After 3-18 months following-up,the appearance and function were satisfactory. Conclusion The distribution of the palmar digital veins was in some pattern.Anastomosis of the superficial palmar digital veins can not only improve the success rate of the fingertip replantation,but also avoid the complications of the other venous drainage methods.
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Objective To report the anatomical study and clinical application of the free peroneal perforator-based sural neurofasciocutaneous flap. Methods Latex injection studies were performed on 10 adult cadaveric lower limbs. The presence, prevalence, and location of the peronel perforators that were suitable for vascular anastomosis were documented. From April 2007 to January 2010, 6 patients with large soft tissue defects in the upper limb underwent the reconstructive procedures with the free peroneal perforatorbased sural neurofasciocutaneous flap transfers. 4 patients were men and 2 were women, with the age ranging from 19 to 60 years. The causes of the injury included 4 motor vehicle accidents, and 2 massive machinery trauma. The area of the flap ranged from 16 cm × 8 cm to 30 cm × 10 cm, and the length of the perforator pedicle was 4 cm to 6 cm. Results A mean of 5.3 perforating vessels of the peroneal artery was noted in the specimen. The peroneal perforator that located at the junction of the middle and lower thirds of the fibula was found suitable for microsurgery, with the external diameter and the length being (1.21 ± 0.13)mm and (4.6 ± 0.8)cm respectively. All the 6 flaps survived completely without necrosis. No severe venous congestion was observed. All the patients were satisfied with the aesthetic outcome postoperatively at 3 to 12 months'follow-up. There were no major donor site complications. Conclusion The free sural neurofasciocutaneous flap based on a single peroneal perforator is an excellent tool for reconstruction of extensive soft tissue defects in the upper limb.
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Objective To discuss the donor site complications and the treatments of the anterolateral thiIgh flap transfer. Methods From July 1988 to July 2007,427 patients treated by anterolateral thish flap surgery were followed up.The donor sites were treated by several methods,including direct closure(49 cases),skin graft transfer(258 cases),anterolateral fascial flap transfer(27 cases),musculocutaneous flap and skin sraft transfer(65 cases), superficial epigastric artery flap transfer(28cases).The area of these flaps ranged from 30 cm×10 cm to 18 cm×7 cm.and the average follow-up was 21 months(ranged from 8 months to 54 months). All the data was analyzed by SPSS for Windows,Version 11.0.1. Results Short-term complications(within 4 weeks)and long-term complications (over 6months)were observed.The former included skin necrosis(5.4%),wound infection(4.0%),and necrosis of rectus femoris(0.5%); and the latter included chronic ulcer(4.9%), aerious scar(6.6%), serious discomfortable(5.9%),and quadriceps femoris disfunction(3.7%).The statistical results showed that there was a close correlation between the donor site morbidity and the surgical methods. Conclusion The complications of the donor site cannot be ignored.Protect the soft-tissue of the donor site carefully,design all appropriate operation plan.Careful preoperative examination and local flap transplantation can effectively decrease the rate of complications.
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<p><b>OBJECTIVE</b>To report the operative technique and clinical application of the neurocutaneous flap with anterior cutaneous branch of the femoral nerve supplied by the perforator of saphenous artery.</p><p><b>METHODS</b>The reverse neurocutaneous flap with anterior cutaneous branch of the femoral nerve supplied by the perforator of saphenous artery was used for repairing the defect around knee or at the upper pad of leg. Since Oct. 2005, 16 cases were treated. The flap size ranged from 15 cm x 7 cm to 30 cm x 15 cm. Flap rotation angle ranged from l00 degrees to 180 degrees.</p><p><b>RESULTS</b>13 flaps survived completely. Flap necrosis happened at the 1/7 - 1/5 distal end of the 3 flaps, which healed with dressing or local flap advancement. The patients were followed up for 6 to 24 months with satisfactory functional and cosmetic results. There was no morbidity at the donor site.</p><p><b>CONCLUSION</b>The flap has the advantages of both the perforator flap and the neurocutaneous flap. The size of the neurocutaneous flap with the anterior cutaneous branch of the femoral nerve can be enlarged for the large defect at lower extremity.</p>
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Adolescent , Adult , Female , Humans , Male , Middle Aged , Young Adult , Arteries , General Surgery , Femoral Nerve , General Surgery , Skin Transplantation , Methods , Surgical FlapsABSTRACT
<p><b>OBJECTIVE</b>To detect the RNA of severe acute respiratory syndrome virus (SARS-CoV) by using reverse transcription polymerase chain reaction (RT-PCR) targeted for a two loci and a modified nested real-time RT-PCR as to improving the reliability and sensitivity of tests.</p><p><b>METHODS</b>A nested RT-PCR was used for detecting one fragment of SARS-CoV RNA in oropharyngeal swabs from 3 SARS probable patients, 4 SARS suspect patients and other 27 patients with fever in Hangzhou, and the nested RT-PCR product from one SARS probable patient was sequenced. Meanwhile in these 3 SARS probable patients, other three RT-PCR methods, including a hemi-nested RT-PCR targeted for another fragment of SARS-CoV RNA, a real-time RT-PCR and a modified nested real-time RT-PCR, were employed to detect SARS-CoV RNA.</p><p><b>RESULTS</b>Two positives were found in the 3 SARS probable patients, and none positive in 4 SARS suspect patients and other 27 patients with fever, using the nested RT-PCR. The sequence of the nested RT-PCR product from one SARS probable patient was identified with the counterpart of SARS-CoV genomes published in public database. The results of the hemi-nested RT-PCR, the real-time RT-PCR and the modified nested real-time RT-PCR in the 3 SARS patients were consistent with the one of the nested RT-PCR. During detecting specimen with low copies of RNA, a weak positive signal was produced after about 35 cycles in the real-time RT-PCR, but a strong positive signal was found only after 10 cycles in the modified nested real-time RT-PCR.</p><p><b>CONCLUSION</b>It might improve the reliability of test by employing RT-PCR targeted for two or more fragments in SARS-CoV genome. The modified nested real-time RT-PCR might have higher sensitivity than the routine real-time RT-PCR.</p>
Subject(s)
Adolescent , Adult , Aged , Aged, 80 and over , Humans , Middle Aged , Young Adult , Base Sequence , RNA, Viral , Genetics , Metabolism , Reproducibility of Results , Reverse Transcriptase Polymerase Chain Reaction , Methods , Severe acute respiratory syndrome-related coronavirus , Genetics , Sensitivity and Specificity , Severe Acute Respiratory Syndrome , Diagnosis , VirologyABSTRACT
<p><b>OBJECTIVE</b>To explore the possible factors causing the increase of rabies cases and to provide references for the development of related prevention and control strategy in Hunan.</p><p><b>METHODS</b>Data was collected and analyzed on epidemic situation of rabies in the past ten years, and studies were carried in some counties.</p><p><b>RESULTS</b>From 1994 to 1999, the annual cases sustained between 17 and 78. However, the number of cases has continued to increase since 2000. The annual reported cases in 2001 and 2002 were 311 and 313, and accounted for 34.8% and 30.0% of the total cases in the whole country respectively. The epidemic areas were mainly located in the south and middle parts of Hunan. Furthermore, the epidemic areas had been expanding. In 1997, human rabies cases were only reported in 7 cities but increased to 12 cities in 2004.</p><p><b>CONCLUSION</b>Factors as increasing number but low inoculating rate to dogs, and incorrect treatment on the wounds etc. might have served important roles for the recurrence of rabies. Hence, it is necessary to take comprehensive preventive measures to control and prevent the epidemics of rabies in Hunan.</p>