Objective To investigate the effect of sevoflurane preconditioning on CCAAT/enhancer binding protein (C/EBP) homologous protein (CHOP) expression in the cerebral cortex after focal cerebral ischemiareperfusion (I/R) injury in rats and the mechanism. Methods Thirty-six male SD rats weighing 250-280 g were randomly divided into 3 groups ( n = 12 each) : sham operation group (group S) , focal cerebral I/R group (group I/R) and sevoflurane preconditioning group (group Sevo-pc). The animals were anesthetized with intraperitoneal chloral hydrate 300 mg/kg. In groups I/R and Sevo-pc, focal cerebral ischemia was induced by middle cerebral artery occlusion using a nylon thread with rounded tip inserted into the right internal carotid artery and advanced cranially until resistance was met. The occlusion was maintained for 1 h followed by 24 h reperfusion. Group Sevo-pc inhaled 2.7% sevoflurane for 1 h before ischemia. Neurological deficits were assessed and scored at the end of 24 h reperfusion and then the rats were decapitated. Their brains were immediately removed. The cerebral infarct size was determined by TTC staining. The CHOP expression in the ischemic cerebral cortex was determined by immunohistochemistry. The number of apoptotic neurons was counted using TUNEL. Results The neurological deficit scores were significantly higher, the cerebral infarct size was significantly larger, and the CHOP expression and the number of apoptotic neurons were significantly higher in groups I/R and Sevo-pc than in group S ( P ＜ 0.01) . The neurological deficit scores were significantly lower, the cerebral infarct size was significantly smaller, and the CHOP expression and the number of apoptosis neurons were significantly lower in group Sevo-pc than in group I/R ( P ＜ 0.05 or 0.01) . Conclusion Sevoflurane preconditioning may protect the brain against focal cerebral I/R injury by down-regulating CHOP expression in the cerebral cortex in rats.

OBJECTIVETo explore the clinical effects of relaying reversed perforator flaps in repairing skin and soft tissue defects at distal end of finger and donor site.

METHODSSeventeen patients (17 fingers) with skin and soft tissue defects at distal end of finger were hospitalized from June 2011 to June 2013. The reversed digital artery perforator flap with branch of digital nerve was used to repair the defect. The first donor site was repaired by dorsal metacarpal artery perforator flap; the second donor site was closed by suturing. The area of skin defect at distal end of finger ranged from 2.0 cm x 1.5 cm to 3.0 cm x 2.0 cm, and the area of digital artery perforator flap and dorsal metacarpal artery perforator flap ranged from 2.2 cm x 1.5 cm to 3.6 cm x 2.5 cm and 2.5 cm x 2.0 cm to 4.2 cm x 3.0 cm, respectively.

RESULTSAll the 34 flaps survived completely. Cyanosis and partial necrosis of the epidermis appeared in 1 flap, which was healed after dressing change. All the patients were followed up for 1 to 18 months, with mean time of 8 months. The color, texture and appearance of flaps were satisfactory. There was no depression or breakdown in the first donor sites. Some linear scars appeared in the second donor sites, but they did not affect the general appearance. The donor sites at joint or tendon did not affect the joint activity after healing. The results of function evaluation of range of active movement of the fingers were excellent in 15 cases and good in 2 cases. The results of sensation of the flaps were S3 in 1 finger, S4 in 2 fingers, and S5 in 14 fingers. The distance of two-point discrimination of flaps ranged from 5 to 7 mm, with mean distance of 6 mm.

CONCLUSIONSRelaying reversed perforator flap, with reliable blood supply and both donor sites in the hand, can improve the appearance and function of the first donor site as well as repair skin and soft tissue defects at distal end of finger.

Cicatrix ; Depression ; Epidermis ; Extremities ; Finger Injuries ; surgery ; Humans ; Perforator Flap ; Reconstructive Surgical Procedures ; methods ; Skin ; Skin Transplantation ; methods ; Soft Tissue Injuries ; surgery ; Surgical Flaps ; blood supply ; Sutures ; Tendons ; Treatment Outcome ; Wound Healing

OBJECTIVETo study the expressions of Notch1-4 gene in human keloid-derived mesenchymal-like stem cells, and to explore the Notch signaling pathway's role in the formation of keloid.

METHODSKeloid samples were collected to harvest human keloid-derived mesenchymal-like stem cells through two-step enzymatic dissociation method. By flow cytometry, cell phenotype of primary and P3 generation were analyzed. By immunocytochemistry, the expressions of Oct4, vimentin and CK19 were examined. Keloid-derived mesenchymal-like stem cells were induced into osteoblasts in vitro and calcium deposition was detected by Alizarin red S stain. Realtime polymerase chain reaction (RT-PCR) was used to detect the expressions of Notch1-4 mRNA in keloid-derived mesenchymal-like stem cells.

RESULTSFlow cytometry showed that keloid-derived mesenchymal-like stem cells of primary and P3 generation highly expressed CD29, CD44, CD90 from the typical MSC phenotype marker, but they failed to express HSC phenotype markers, such as CD34 and CD45. The results of immunocytochemistry showed that Oct4 from pluripotent stem cell markers and vimentin from mesenchymal cell markers was positive and CK19 from epithelial cell markers was negative. After induced differentiation into osteoblasts in vitro after 21 day, calcium nodules could be seen clearly; Notch1-4 gene were expressed in keloid-derived mesenchymal-like stem cells through RT-PCR. The relative quantitative of Notch2, Notch3 gene were higher than Notch1, Notch4 gene (P < 0.05).

CONCLUSIONSThe expression difference of different subtypes from Notch gene in human keloid-derived mesenchymal-like stem ceils may be related to self-renewal, proliferation, differentiation, and participate in the formation of keloid.

Adolescent ; Cells, Cultured ; Child ; Female ; Humans ; Keloid ; metabolism ; pathology ; Male ; Mesenchymal Stromal Cells ; metabolism ; Receptors, Notch ; metabolism

Objective: To investigate the effect of human adipose-derived stem cells (hADSCs) on pressure ulcers in mouse. Methods: The subcutaneous adipose tissue from voluntary donation was harvested. Then the hADSCs were isolated and cultured by mechanical isolation combined with typeⅠcollagenase digestion. The 3rd generation cells were identified by osteogenic, adipogenic, chondrogenic differentiations and flow cytometry. The platelet rich plasma (PRP) from peripheral blood donated by healthy volunteers was prepared by centrifugation. The pressure ulcer model was established in 45 C57BL/6 mice by two magnets pressurized the back skin, and randomly divided into 3 groups ( n=15). The wounds were injected with 100 μL of hADSCs (1×10 6 cells) transfected with a green fluorescent protein (GFP)-carrying virus, 100 μL human PRP, and 100 μL PBS in hADSCs group, PRP group, and control group, respectively. The wound healing was observed after injection. The wound healing rate was calculated on the 5th, 9th, and 13th days. On the 5th, 11th, and 21st day, the specimens were stained with HE staing, Masson staining, and CD31 and S100 immunohistochemical staining to observe the vascular and nerve regeneration of the wound. In hADSCs group, fluorescence tracer method was used to observe the colonization and survival of the cells on the 11th day. Results: The cultured cells were identified as hADSCs by induced differentiation and flow cytometry. The platelet counting was significantly higher in PRP group than in normal peripheral blood group ( t=5.781, P=0.029). General observation showed that the wound healing in hADSCs group was superior to those in PRP group and control group after injection. On the 5th, 9th, and 13th days, the wound healing rate in hADSCs group was significantly higher than those in PRP group and control group ( P<0.05). Histological observation showed that compared with PRP group and control group, inflammatory cell infiltration and inflammatory reaction were significantly reduced in hADSCs group, collagen deposition was significantly increased, and skin appendage regeneration was seen on the 21st day; at each time point, the expression of collagen was significantly higher in hADSCs group than in PRP group and control group ( P<0.05). Immunohistochemical staining showed that the number of neovascularization and the percentage of S100-positive cells in hADSCs group were significantly better than those in PRP group and control group on the 5th, 9th, and 13th days ( P<0.05). Fluorescent tracer method showed that the hADSCs could colonize the wound and survive during 11 days after injection. Conclusion: Local transplantation of hADSCs can accelerate healing of pressure ulcer wounds in mice and improve healing quality by promoting revascularization and nerve regeneration.

OBJECTIVETo explore the curative effect of reverse bi-pedicle posterior interosseous artery perforator flap in repairing skin and soft tissue defects on the wrist.

METHODSSeven patients with soft tissue defects on the wrist, including simple skin and soft tissue defects in 4 cases and skin and soft tissue defects combined with radial tendon injury in 3 cases, were hospitalized from December 2010 to March 2012. The area of skin defect on the volar side of the wrist ranged from 4.8 cm x 4.0 cm to 6.2 cm x 4.5 cm, while that on the dorsal side ranged from 3.5 cm x 3.2 cm to 6. 5 cm x 5.4 cm. These wounds were respectively caused by traffic injury (3 cases), reamer injury (2 cases), burn (1 case), and tumor resection (1 case). Reverse bi-pedicle posterior interosseous artery perforator flaps were used to repair these defects, with area of one pedicle ranging from 2.5 cm x 2.0 cm to 3.5 cm x 2.5 cm and the area of the other pedicle ranging from 2.5 cm x 2.5 cm to 4.0 cm x 3.0 cm. The donor sites were closed by suturing.

RESULTSAll flaps survived completely. Patients were followed up for 6 to 36 months. The color, texture, and appearance of all flaps were satisfactory. At last follow-up, distances of two-point discrimination of flaps ranged from 9 to 13 mm. The dorsal extension and palmar flexion functions of wrist were satisfactory. The results of function evaluation of 7 wrists were excellent in 6 cases and good in 1 case according to the tentative standards for the evaluation of upper extremity function of Society of Hand Surgery of Chinese Medical Association. A linear scar was formed at the donor site.

CONCLUSIONSThe reverse bi-pedicle posterior interosseous artery perforator flap, with advantages of flexible design, easy to achieve, less injury to donor site, and reliable blood supply, etc., is another choice for repairing skin and soft tissue defects over the wrist.

Burns ; Cicatrix ; Humans ; Perforator Flap ; blood supply ; Range of Motion, Articular ; Reconstructive Surgical Procedures ; methods ; Skin ; Skin Transplantation ; methods ; Soft Tissue Injuries ; surgery ; Surgical Flaps ; blood supply ; Tendon Injuries ; Treatment Outcome ; Ulnar Artery ; Wound Healing ; Wrist Injuries ; surgery

Adipose-derived stem cells (ADSCs) are adult mesenchymal stem cells in adipose tissue with self-renewal and multi-directional differentiation potential. The application of ADSCs in the treatment of wounds has achieved good results. Because of its extensive sources, high content in vivo, low immunogenicity, slight injury to body when obtained, the clinical application prospect of ADSCs is promising. The reasons why diabetic wound is difficult to heal may be closely related to the increase of advanced glycation end products, long-term chronic inflammatory response, and peripheral neurologic dysfunction. The abnormal internal environment of diabetic patients can affect the biological function of ADSCs, which further affects wound healing. This article reviews the general feature, differentiation, proliferation, migration, secretion, and pro-angiogenic function of diabetic ADSCs.

Objective:To investigate the clinical effect of the anterolateral thigh perforator chimeric flap in the treatment of the wound of diabetic foot ulcer (DFU) .Methods:From January, 2018 to December, 2019, 14 cases wound of DFU of type II diabetic were treated by anterolateral thigh chimeric perforator flap. The patients were 10 males and 4 females, at 49 to 58 years old. Of the 14 patients, 10 with simple peripheral neuropathy, 4 with peripheral neuropathy complicated with vascular disease, and none with single vascular disease. With strict control of patients' blood glucose, antibiotics blended bone cement was applied or filled onto grade 2 or higher grade Wagner's DFU after debridement. In addition, the anterolateral thigh chimeric perforator flap was transferred 2 to 3 weeks later. The size of flap was 8 cm×3 cm-27 cm×7 cm. Regular followed-up were made after surgery.Results:Thirteen flaps survived in one stage after surgery. The other 1 flap had venous vascular crisis, and survived completely after active exploration. The patients were followed-up for 6-12 months. All the flaps survived well in good shape and texture. The donor and recipient areas healed well. The functional recoveries of the DF were satisfactory.Conclusion:Application of anterolateral thigh perforator chimeric flap in repair of the refractory wound of DF achieves a good clinical outcome and effectively improves the life quality of patients.

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.

To investigate the effects of adipose-derived mesenchymal stem cells (AMSCs) from type 2 diabetes mellitus patients on wound healing of pressure ulcers in mice.Methods: (1) In September 2016, the subcutaneous adipose tissue of a 60-year-old woman with type 2 diabetes mellitus was harvested, and then AMSCs were extracted by collagenase digestion and cultured. The third passage of cells were used for subsequent experiments. The morphology of cells was observed, and their osteogenic, chondrogenic, and adipogenic differentiation abilities were identified. The expressions of cell surface markers CD90, CD105, CD73, and CD34 were detected by flow cytometer (n=3). (2) Sixteen female C57BL/6 wild-type mice aged 6-8 weeks were selected, and one pressure ulcer wound was created on each side of the spine of each mouse by pressing the skin with two magnets. The two wounds of each mouse were paired and divided into diabetic AMSCs group and negative control group, injected with 100 μL phosphate buffer solution (PBS) containing green fluorescent protein-labeled AMSCs (1×10⁶ cells) and 100 μL PBS, respectively. The wound healing status of the two groups within post injection day (PID) 21 was observed, and their wound healing rates on PID 5, 13, and 17 were calculated. Three mice were sacrificed on PID 11 and 21, respectively, and tissue of three wounds was harvested from each group. The skin structure was observed by hematoxylin-eosin staining, the collagen deposition was evaluated by Masson staining, and the positive expression of CD31, i. e., the number of new blood vessels was counted by immunohistochemistry. Wound tissue samples of two groups prepared on PID 21 as above-mentioned were harvested, and the positive cell rate of S100, representing the regeneration of Schwann cells, was detected by immunohistochemistry. Wound tissue samples of diabetic AMSCs group prepared on PID 11 as above-mentioned were harvested, and the colonization of AMSCs was observed by fluorescence tracer method. Data were processed with paired t test and Bonferroni correction. Results: (1) The third passage of cells isolated and cultured from the subcutaneous adipose tissue of a type 2 diabetes mellitus patient grew adherently to the wall in a long spindle and vortex-like manner. After induction, the cells showed osteogenic, chondrogenic, and lipogenic differentiation abilities. The positive expression rates of CD90, CD105, and CD73 on the cell surface were higher than 90.00%, and the expression rate of CD34 was 0.46%. The cells were identified as AMSCs. (2) The mice wounds of diabetic AMSCs group healed quickly, and all the wounds healed completely on PID 17, while the mice wounds in negative control group were not completely closed at this time, and there was still scab on the surface. On PID 5, 13, and 17, the healing rates of mice wounds of diabetic AMSCs group were (35.6±6.5)%, (87.1±2.5)%, and 100.0%, respectively, significantly higher than (19.8±7.2)%, (66.2±5.2)%, and (86.9±5.3)% of negative control group (t=6.49, 14.31, 9.73, P<0.05). Compared with that of negative control group, the inflammatory cell infiltration was reduced in mice wounds tissue of diabetic AMSCs group on PID 11, and thicker epidermis and dermis as well as regenerated skin appendages were observed on PID 21. On PID 11 and 21, the collagen percentages of mice wounds tissue in diabetic AMSCs group was (48.3±1.3)% and (54.1±1.7)%, respectively, significantly higher than (41.4±1.7)% and (50.3±1.2)% of negative control group (t=6.98, 3.99, P<0.01). On PID 11 and 21, the numbers of new blood vessels in mice wounds tissue of diabetic AMSCs group were 17.2±1.3 and 18.0±2.1, respectively, significantly more than 8.0±1.4 and 14.0±1.5 of negative control group (t=10.69, 3.38, P<0.01). On PID 21, the S100 positive cell percentage in mice wounds tissue of diabetic AMSCs group was (1.76±0.12)%, significantly higher than (0.55±0.03)% of negative control group (t=21.68, P<0.001). On PID 11, the colonization of AMSCs in mice wounds tissue of diabetic AMSCs group was observed. Conclusions Transplantation of AMSCs from type 2 diabetic mellitus patients can accelerate wound healing of pressure ulcers in mice by promoting angiogenesis, collagen deposition, and Schwann cell regeneration.

Objective: To explore the effects of local transplantation of autologous adipose-derived stromal vascular fraction (SVF) on the hyperplastic scar (HS) formation in rabbit ears and the mechanism. Methods: Twenty-four New Zealand white rabbits were used to reproduce HSs by making four full-thickness skin defect wounds with a diameter of 1 cm on the ventral surface of left ear of each rabbit. Wound epithelization and local-tissue proliferation were observed, and wound healing (complete epithelization) time and formation time of HS were recorded. The 24 rabbits were divided into SVF group, pure DMEM group, and pure HS group according to the random number table, with 8 rabbits and 32 wounds in each group. On post injury day (PID) 25 (after the complete epithelization of wounds), 0.2 mL of low glucose DMEM medium containing CM-Dil labeled autologous SVF was injected into HSs of rabbits in SVF group, while the same amount of low glucose DMEM medium was injected into HSs of rabbits in pure DMEM group. The frequency of injection was once every 5 days, totally for 3 times. HSs of rabbits in pure HS group did not receive any treatment. On PID 40, HSs of rabbits′ ears in each group were harvested, then the histological form was observed by hematoxylin and eosin staining, the arrangement of collagen in HS was observed by Van Gieson staining, the distribution of CM-Dil-labeled SVF in the HS was observed with fluorescence microscope, and the mRNA expression and the protein expression of transforming growth factor β₁ (TGF-β₁), Smad3, and Smad7 in HS were determined by real-time fluorescent quantitative reverse transcription-polymerase chain reaction and Western blotting, respectively. Data were processed with one-way analysis of variance and Tukey test. Results: (1) Complete epithelization time of wounds of rabbits′ ears was (20.0±2.0) d post injury, and HSs were formed on PID 25. On PID 40, HSs of rabbits′ ears in pure DMEM group and pure HS group were still in hyperplasia, while those in SVF group became smaller, flat, soft, and light colored. (2) On PID 40, compared with those in pure DMEM group and pure HS group, the number of epithelium foot like structures was more and the amount of inflammatory cells was less. The collagen of HSs of rabbits′ ears in SVF group was arranged more regularly with broader gap between collagens. (3) On PID 40, CM-Dil-labeled SVF could still be observed in the HSs of rabbits′ ears in SVF group. (4) On PID 40, compared with those in pure DMEM group and pure HS group, the mRNA expressions of TGF-β₁ and Smad3 in the HSs of rabbits′ ears in SVF group were significantly down-regulated (P<0.05), while the mRNA expression of Smad7 was significantly up-regulated (P<0.05). There were no significant differences in the mRNA expressions of TGF-β₁, Smad3, and Smad7 in the HSs of rabbits′ ears between pure DMEM group and pure HS group (P>0.05). (5) On PID 40, compared with those in pure DMEM group (0.74±0.03, 0.73±0.10, 0.54±0.09) and pure HS group (0.72±0.08, 0.71±0.12, 0.53±0.06), the protein expressions of TGF-β₁ and Smad3 in the HSs of rabbits′ ears in SVF group (0.57±0.06, 0.42±0.09) were significantly down-regulated (P<0.05), while the protein expression of Smad7 (0.71±0.05) was significantly up-regulated (P<0.05). The protein expressions of TGF-β₁, Smad3, and Smad7 in the HSs of rabbits′ ears in pure DMEM group and pure HS group were close (P>0.05). Conclusions Autologous SVF transplantation can inhibit the formation of HS in the early stage of scar formation of rabbit, the mechanism may be related to the TGF-β₁/Smad signaling pathway.