Objective:To explore the method and clinical effect of repairing neck radiation ulcer with superficial cervical artery flap.Methods:January 2016 to June 2019, 11 cases of neck radiation ulcer were repaired with superficial cervical artery flap. The ulcer occurred 13.4 years after radiotherapy on average, with an area between 1 cm×2 cm and 3 cm×7 cm, extensive fibrosis.After pathological examination, surgical removal of ulcer and surrounding fibrotic tissue, and avoid injuring the neck’s main blood vessels. Wound size after debridement: 6 cm×9 cm-8 cm×13 cm. The flap rotation point is 4-5 cm beside the spine on the acromion level.The flaps were designed along the skin branch of the superficial cervical artery which located by the Doppler blood flow detector.The distance from the rotation point to the flap’s proximal edge is about 2 cm longer than that of the wound’s proximal edge.The size of the flap is about 2 cm larger than the wound.Then the flaps were excised from far to near after exposing the pedicle. The flap was then transferred to the neck to repair the defect formed by the radiation ulcer’s excision. If the tension is low, the donor sites were dissociated about 2-3 cm, then sutureddirectly after reducing tension.If the tension is high, part of the donor site was sutured to reduce the area and then grafted with medium-thickness skin.The postoperative changes of the patient were carefully observed.Among them, 7 cases underwent one-stage operation, 4 cases underwent two-stage operation after the donor area’s pre-expansion.Results:All patients with radiation ulcers were healed completely.The appearance and function of the operation area were good.Followed up for 6-24 months, no recurrences of radiation ulcer were observed. Among them, the flaps of 10 cases survived completely, and the wound healed in one stage.The distal area of the flap necrosed in one case, and repaired by dressing change and skin grafting.Conclusions:Radiation ulcer of the neck is a serious long-term complication after radiotherapy. Once it occurs, it is difficult to heal by conservative treatment. Skin grafting is difficult to survive as well. The superficial cervical artery flap has a constant and abundant blood supply and a hidden donor area, which is a feasible method for the treatment of radiation ulcer of the neck and reconstruction of the function.

Objective:To explore the clinical effects of flaps with cervical cutaneous branch of transverse cervical artery in repairing neck radiation ulcer.Methods:The retrospective observational research was conducted. From January 2016 to December 2019, 8 cases with neck radiation ulcer were admitted to Guangzhou Red Cross Hospital of Jinan University and repaired with flaps based on cervical cutaneous branch of transverse cervical artery. There were 6 males and 2 females, aged 52-75 years. The ulcers occurred 14.5 years after radiotherapy on average, with ulcer areas of 5.0 cm×3.0 cm-7.0 cm×6.0 cm on admission. The wound areas were ranged from 6.0 cm×5.0 cm to 13.0 cm×6.5 cm after ulcers and fibrotic tissue around were resected. Seven cases underwent resection of flaps and wound repair operation on the first stage, and the other 1 case underwent pre-expansion of flap donor area on the first stage and resection of flap and wound repair operation on the second stage, with flap sizes of 8.0 cm×7.0 cm-15.0 cm×8.5 cm. The wounds in the donor areas of flaps in 7 patients were sutured directly, and the wound in the donor area of flap in the other 1 patient was repaired with thin split-thickness skin graft from thigh after being sutured partially. The preoperative ulcer tissue was collected for pathological examination, and the postoperative survival of the flaps and healing of the flap donor areas were observed. The flaps, the recurrence of the neck ulcers and neck function, and the scar hyperplasia in the donor areas of flaps were observed during follow-up.Results:Preoperative pathological examination of ulcer tissue showed that full-thickness necrosis occurred in ulcer skin, obvious fibrotic tissue hyperplasia, collagenization, and small-scale calcification in the base and surrounding tissue of the ulcers, and a large amount of chronic inflammatory cells and a small amount of acute inflammatory cells infiltration were observed in intercellular substance, which excluded the recurrence of tumor. All the flaps in 8 cases survived, the wounds were repaired effectively, and the postoperative donor areas of flaps healed well. During postoperative follow-up of 6-24 months, the flaps had good appearances without recurrence of ulcer, the movement function of neck was significantly improved, and no obvious scar hyperplasia was observed in the donor areas of flaps.Conclusions:Radiation ulcer in the neck is a serious long-term complication of neck after radiotherapy, which is difficult to heal with conservative treatment. The flap with cervical cutaneous branch of transverse cervical artery is close to the neck, with rich blood supply, constant anatomy, and is easy to cut. Neck radiation ulcers treated with the flaps showed good wound healing and improved functions, with no obvious scar hyperplasia.

Objective:To explore the method and clinical effect of repairing neck radiation ulcer with superficial cervical artery flap.Methods:January 2016 to June 2019, 11 cases of neck radiation ulcer were repaired with superficial cervical artery flap. The ulcer occurred 13.4 years after radiotherapy on average, with an area between 1 cm×2 cm and 3 cm×7 cm, extensive fibrosis.After pathological examination, surgical removal of ulcer and surrounding fibrotic tissue, and avoid injuring the neck’s main blood vessels. Wound size after debridement: 6 cm×9 cm-8 cm×13 cm. The flap rotation point is 4-5 cm beside the spine on the acromion level.The flaps were designed along the skin branch of the superficial cervical artery which located by the Doppler blood flow detector.The distance from the rotation point to the flap’s proximal edge is about 2 cm longer than that of the wound’s proximal edge.The size of the flap is about 2 cm larger than the wound.Then the flaps were excised from far to near after exposing the pedicle. The flap was then transferred to the neck to repair the defect formed by the radiation ulcer’s excision. If the tension is low, the donor sites were dissociated about 2-3 cm, then sutureddirectly after reducing tension.If the tension is high, part of the donor site was sutured to reduce the area and then grafted with medium-thickness skin.The postoperative changes of the patient were carefully observed.Among them, 7 cases underwent one-stage operation, 4 cases underwent two-stage operation after the donor area’s pre-expansion.Results:All patients with radiation ulcers were healed completely.The appearance and function of the operation area were good.Followed up for 6-24 months, no recurrences of radiation ulcer were observed. Among them, the flaps of 10 cases survived completely, and the wound healed in one stage.The distal area of the flap necrosed in one case, and repaired by dressing change and skin grafting.Conclusions:Radiation ulcer of the neck is a serious long-term complication after radiotherapy. Once it occurs, it is difficult to heal by conservative treatment. Skin grafting is difficult to survive as well. The superficial cervical artery flap has a constant and abundant blood supply and a hidden donor area, which is a feasible method for the treatment of radiation ulcer of the neck and reconstruction of the function.

Objective To investigate the effects of quercetin on glial scar formation and axonal regeneration after spinal cord injury (SCI) and its association with the p38 mitogen activated protein kinase (MAPK) signal pathway.Methods 128 female Sprague-Dawley (SD) rats were randomly divided into a control group (SCI + saline),an intervention group (SCI + quercetin + anisomycin),a treatment group (SCI + quercetin) and a sham-operation group (n =32).Basso Beattie Bresnahan (BBB) assessment and footprint analysis of the hind limb were performed on days 1,3,7,14,21 and 28 postoperation in each group.The expression levels of p38MAPK,phosphorylation p38MAPK,glial fibrillary acidic protein (GFAP) and neurofilament protein-200 (NF-200) were detected by Western blot.The numbers of GFAP and NF-200 positive staining cells in the injured spinal cord in each group were detected by immunohistochemistry.Results The BBB scores in the treatment group were significantly higher than in the intervention and control groups at each time point after SCI except on day 3 postoperation (P ＜ 0.05).The expression levels of phosphorylation p38MAPK protein in each SCI group were significantly higher than in the sham-operation group on days 3 and 7 postoperation (P ＜ 0.05).The expression levels of phosphorylation p38MAPK protein in the treatment group were significantly lower than in the control and intervention groups on days 3,7 and 14 postoperation (P ＜ 0.05),but there was no significant difference on day 28 postoperation among all the groups (P ＞ 0.05).The numbers of NF-200 and GFAP positive staining cells were significantly greater than in the sham-operation group at each time point postoperation (P ＜ 0.05);the NF-200 positive staining cells in the treatment group were significantly increased in comparison with the control and intervention groups (P ＜ 0.05);the GFAP positive staining cells in the treatment group were significandy fewer than in the control and intervention groups on days 7,14 and 28 postoperation (P ＜ 0.05).Conclusions Quercetin may have protective effects against acute SCI by decreasing glial scar formation,increasing axonal regeneration and promoting recovery of locomotor and nerve function in rats.The effects may be correlated with inhibition of the p38MAPK signal pathway.

Previous studies have demonstrated that integrin-linked kinases (ILKs) are abundantly expressed in extracellular matrix (ECM) riche dermis, hair follicles, and basal cells of epidermis. ILKs are not only essential for the maintenance of skin structure, but also play important roles in wound healing. ILKs can promote the formation of granulation tissue by stimulating the proliferation of fibroblasts and secretion of ECM, accelerate wound contraction by inducing the differentiation of fibroblasts to myofibroblasts, and boost reepithelization by promoting proliferation, migration, and differentiation of keratinocytes and follicle epidermal stem cells.

Integrin-linked kinase (ILK) has been found for twenty years, and its biological characteristics have been extensively studied by multi-discipline. At present, studies of ILK are mainly focused on its roles in angiogenesis, tumor formation, and tissue fibrosis, etc. In recent years, the regulation effect of ILK in angiogenesis attracts attention of researchers. The studies showed that ILK can stimulate the secretion of angiogenic factor, promote the proliferation and migration of endothelial cells and inhibit their apoptosis, and therefore play an important role in the regulation of angiogenesis. Further research on molecular mechanism about the role of ILK playing in angiogenesis may provide an effective method for the treatment of some diseases.

OBJECTIVETo investigate the role of integrin-linked kinase (ILK) signaling pathway in the skin lesions and wound healing in diabetic rats.

METHODSThirty-six SD rats were divided into diabetic wound group (D) and non-diabetic wound group (N) according to the random number table, with 18 rats in each group. 10 g/L streptozocin (60 mg/kg) was intraperitoneally injected in rats in group D, while the rats in group N were given same quantity of sodium citrate buffer. Two weeks after successful reproduction of diabetic model of rats in group D, two full-thickness skin of an area of 2 cm × 2 cm was resected on both sides of back of rats in the two groups. Wounds of three rats of each group were photographed and examined on post injury day (PID) 1, 3, 7, 10, 14, and 21, and the wound healing rates were calculated. The non-injured skin and wound tissue (central part) on back of three rats of the rest 15 rats in the two groups were harvested on PID 3, 7, 10, 14, and 21, respectively. Morphology of the non-injured skin tissue was observed with HE staining, and the thickness of full-thickness skin and epidermis were measured. The mRNA expression levels of ILK, protein kinase B (Akt), and glycogen synthase kinase-3β (GSK-3β) in non-injured skin tissue were determined with real-time fluorescent quantitative RT-PCR. The protein expression levels of ILK, Akt, phosphorylated Akt, GSK-3β, and phosphorylated GSK-3β in non-injured skin tissue, and ILK, phosphorylated Akt in wound tissue were assessed with Western blotting. Data were processed with two independent-sample t test, one-way analysis of variance, SNK test and analysis of variance of factorial design.

RESULTS(1) After injury, the wound scabs of rats in group N were dry, and red granulation tissue with no excretion were seen when the scabs fell off, and the wound healed fast. After injury, excretion under the wound scabs of rats in group D was seen, and the scabs easily fell off with exposure of pink granulation tissue with much excretion, and the wounds healed slowly. Except for PID 3, the wound healing rate of rats in group D was significantly lower than that in group N on other PIDs (with t values from 3.858 to 13.738, P<0.05 or P<0.01). (2) On PID 3, the hair follicles and blood vessels in the non-injured skin tissue of rats in group N were rich, and the epidermis was composed of stratified cells in form of basal cells and keratinocyte, and the hair follicles and blood vessels in the non-injured skin tissue of rats in group D were scarce, and the epidermis was nearly composed of one-layer of cells. The thickness of full-thickness skin and epidermis of non-injured skin tissue of rats in group N was similar from PID 3 to 21, and the thickness of full-thickness skin and epidermis of non-injured skin tissue of rats in group D on PID 3 was respectively (1 074 ± 66) and (15.1 ± 3.8) μm, and they gradually thinned out to (785 ± 122) and (9.7 ± 2.1) μm on PID 21, respectively. The thickness of full-thickness skin and epidermis of non-injured skin tissue of rats in group N were significantly thicker than those in group D on each PID (with t values from 4.620 to 23.549, P values below 0.001). (3) From PID 3 to 21, the mRNA expression levels of ILK and Akt in non-injured skin tissue of rats in group D were significantly lower than those in group N (with t values respectively 4.779 and 3.440, P values below 0.05), the mRNA expression levels of GSK-3β in non-injured skin tissue of rats were similar in two groups (t=0.363, P>0.05). (4) From PID 3 to 21, the protein expression levels of ILK, Akt and phosphorylated Akt in non-injured skin tissue of rats in group D were significantly lower than those in group N (with t values from 2.630 to 6.209, P<0.05 or P<0.01); the protein expression levels of GSK-3β in non-injured skin tissue of rats in two groups were similar (t=0.652, P>0.05); the protein expression level of phosphorylated GSK-3β in non-injured skin tissue of rats in group D was significantly higher than that in group N (t=4.131, P<0.001). The protein expression levels of ILK in wound tissue of rats in two groups were similar on each PID (with t values from 0.381 to 2.440, P values above 0.05). Except for PID 3, the protein expression levels of phosphorylated Akt in wound tissue of rats in group N were significantly higher than that in group D on other PIDs (with t values from 4.091 to 20.555, P<0.05 or P<0.01). From PID 3 to 21, the protein expression levels of ILK in wound tissue and non-injured skin tissue of rats in group N were similar (F=2.522, P>0.05), and the protein expression level of phosphorylated Akt in wound tissue was significantly higher than that in non-injured skin tissue (F=117.329, P<0.001); the protein expression levels of ILK in wound tissue and non-injured skin tissue of rats in group D were similar (F=1.337, P>0.05), and the protein expression level of phosphorylated Akt in wound tissue was significantly higher than that in non-injured skin tissue (F=184.120, P<0.001).

CONCLUSIONSThe skin lesion of diabetic rats may be related to the declined expression levels of ILK, Akt and phosphorylated Akt in the ILK signaling pathway. The refractory healing of wound in diabetic rats may be related to the declined expression level of phosphorylated Akt.

Animals ; Diabetes Mellitus, Experimental ; enzymology ; Glycogen Synthase Kinase 3 ; metabolism ; Glycogen Synthase Kinase 3 beta ; Phosphorylation ; Protein-Serine-Threonine Kinases ; metabolism ; Proto-Oncogene Proteins c-akt ; metabolism ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; Skin ; injuries ; Wound Healing

Objective To investigate the effects of retinoic acid (RA) on the proliferation and apoptosis of mesenchyme and epithelium at the key stages of palatal development,and to determine the embryonic alterations associated with cleft palate.Methods 100 pregnant C57 females (E12) were equally divided into two groups.The experimental group was administered RA once (70 mg/kg) while the control group was administered corn oil only,and the heads of the embryos at E13.5-17.5 were collected and processed to paraffin.Sections were stained with hematoxylin and eosin for the morphological assessment of anterior,posterior,left and right palate,and BrdU and TUNEL assays for the detection of proliferation and survival of palatal mesenchyme and epithelium.Results Simple treatment of RA at 70 mg/kg caused an incidence of 100％ cleft palate.During E13.5-15.5,cell proliferation was significantly promoted on the anterior palatal mesenchyme in the RA group,while no difference for the posterior palatal mesenchyme.Besides,the cell proliferation on anterior and posterior epithelium was comparable between the control and RA group.Less apoptotic cells were observed on epithelium during El3.5-14.5 in the RA group.Conclusions RA induces excessive cell proliferation of palatal mesenchyme,causing abnormal vertical palatal growth and failure of palatal shelves contact and fusion,which finally lead to cleft palate.RA also increases epithelium apoptosis.

Objective To investigate the effects of retinoic acid (RA) on the proliferation and apoptosis of mesenchyme and epithelium at the key stages of palatal development,and to determine the embryonic alterations associated with cleft palate.Methods 100 pregnant C57 females (E12) were equally divided into two groups.The experimental group was administered RA once (70 mg/kg) while the control group was administered corn oil only,and the heads of the embryos at E13.5-17.5 were collected and processed to paraffin.Sections were stained with hematoxylin and eosin for the morphological assessment of anterior,posterior,left and right palate,and BrdU and TUNEL assays for the detection of proliferation and survival of palatal mesenchyme and epithelium.Results Simple treatment of RA at 70 mg/kg caused an incidence of 100％ cleft palate.During E13.5-15.5,cell proliferation was significantly promoted on the anterior palatal mesenchyme in the RA group,while no difference for the posterior palatal mesenchyme.Besides,the cell proliferation on anterior and posterior epithelium was comparable between the control and RA group.Less apoptotic cells were observed on epithelium during El3.5-14.5 in the RA group.Conclusions RA induces excessive cell proliferation of palatal mesenchyme,causing abnormal vertical palatal growth and failure of palatal shelves contact and fusion,which finally lead to cleft palate.RA also increases epithelium apoptosis.

OBJECTIVETo study the role of integrin-linked kinase (ILK) on the proliferation and differentiation of human fibroblast in hypertrophic scar and its effect on the scar formation.

METHODSThe human scar fibroblasts were isolated and cultured in vitro. The cells were divided into 4 groups. (1) control group: only contains DMEM; (2) jetPRIME group: DMEM with 200 microl jetPRIME buffer and 4 microl jetPRIME; (3) ILK siRNA group: DMEM and ILK siRNA; (4) ILK cDNA group: DMEM and ILK cDNA. The cell proliferation was detected by XTT assay and the mRNA and protein expressions of ILK and alpha-SMA were detected by Real-time qPCR and Western blot.

RESULTS(1) XTT results showed that the cellular proliferation level after 48 h in four groups were 0.820 +/- 0.065, 0.873 +/- 0.041, 0.554 +/- 0.013 and 1.296 +/- 0.094, respectively. The cellular proliferation curve showed that the cellular proliferation level was very flat in ILK siRNA group while the cellular proliferation level gradually increased from 12 h. 48 h after transfection, the cellular proliferation level in ILK siRNA group was significant lower than those in other groups (P value were 0.021, 0.034, 0), while the cellular proliferation level in ILK cDNA group was the highest among all 4 groups (P value were 0.017, 0.009, 0). (2) The Real-time qPCR showed that the expressions of ILK mRNA and alpha-SMA mRNA were 0.693 +/- 0.412 and 0.422 +/- 0.037 in control group, were 0.621 +/- 0.183 and 0.388 +/- 0.005 in jetPRIME group, were 0.052 +/- 0.019 and 0.073 +/- 0.023 in ILK siRNA group, were 240.193 +/- 35.170 and 138.056 +/- 24.060 in ILK cDNA group. The expressions of ILK mRNA and alpha-SMA mRNA in ILK siRNA group were significantly lower than those in other three groups (P < 0.05). And the expressions of ILK mRNA and alpha-SMA mRNA in ILK cDNA group were significantly higher than those in other three groups (P < 0.05). (3) The Western blot also showed that the expression of ILK and alpha-SMA proteins were decreased in ILK siRNA group and increased in ILK cDNA group.

CONCLUSIONILK may promote the proliferation and differentiation of human scar fibroblast. It may play an important role in scar formation and contracture.

Actins ; metabolism ; Adolescent ; Adult ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Cicatrix, Hypertrophic ; metabolism ; Female ; Fibroblasts ; cytology ; drug effects ; metabolism ; Humans ; Male ; Middle Aged ; Protein-Serine-Threonine Kinases ; pharmacology ; RNA, Messenger ; genetics ; Transfection ; Young Adult