Objective To investigate treatment effects of correcte d severe blepharoptosis using an orbicularis oculi muscle flap. Meth ods After the lid creased incision, the subtaneous plane was dissect ed between the skin and the orbicularis oculi muscle, reaching as far as the sup erior margin of eyebrow. In a plane between the obicularis and the orbital sept um, the dissection was extended bluntly upward to above the superior orbital rim . to advance the orbicilaris oculi muscle flap to the tarsal plate and to suture the flap with the tarsus directly. Results Based on cri teria of the ″satisfactory″results by Souther, postoperative results of all bl ep haroptosis of twenty-four eyes in fifteen paitents were satisfactory. Conclusion The orbicularis oculi muscle flap for correction of s evere blepharoptosis is simple and reliable technique, which has several advanta ges over the conventional frontalis muscle flap technique, such as single incis ion on the supratarsal fold, the preservation of the function of frontalis mucle , no depression on the forehead, no risk of neurovascular injury and no asymmet rical eyebrows in unilateral ptosis.

Objective To improve the therapeutic effects and to prevent the recurrence of the blepharoptosis. Methods The frontalis muscle was cut to form the ladder shaped frontalis muscle flaps, which was than were transferred under the orbicularis to touch the tarsus with its broad lower part perfectly. The flap was sutured with the tarsus directly. Results All blepharoptosis of twenty four eyes in sixteen patients were treated with excellent results in function and cosmetic appearance. The upper eyelids could be closed effectively. There were no any recurrence and complications in this group. Conclusions The transfer of the ladder shaped frontalis muscle flaps applied in this group benefits the moving of upper eyelid because of its effectiveness in contraction and stable suture with the tarsus. It is more suitable for the treatment of severe blepharoptosis.

OBJECTIVE To observe the bacterial content on the suspension bed sheet in burns intensive care unit.METHODS To detect the bacterial content on the suspension bed sheet and the common sickbed sheet(20 sheets in each group) after sampling them before use and 12 and 24 hours after use and to compare them.RESULTS The bacterial content on two sorts of the sheets before use was accorded with the Ministry of Health standards.but 12 hours after use the significant bacterial content on the common sickbed sheet [(4.9?2.1) CFU/cm2] was much higher than that on the suspension bed [(3.2?1.1) CFU/cm2,P0.01].CONCLUSIONS Using suspension bed could reduce bacterial content on the sheet and ease nurse workload.

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

BACKGROUND: Transforming growth factor-β(TGF-β) is an essential factor for pathological scar formation. Smad protein group is the signal protein of lower reaches of TGF-β receptor. Asiaticoside can inhibit proliferation of fibroblasts and synthesis of collagen to reduce TGF-β expression in the scar.OBJECTIVE: To study the mechanism of asiaticoside on proliferation of scar fibroblasts and phosphorylated Smad2 and Smad7.DESIGN: Controlled study with observation, in which cell was taken as the object.SETTING: Department of burn and plastic surgery of a hospital affiliated to a university.PARTICIPANTS: The experiment was performed in Surgical Laboratory in Sun Yat-sen University from April 2002 to March 2003. The specimens were selected from 6 inpatients receiving plastic operation due to hyperplasic scar including 3 male and 3 female cases aged varied from 1 to 35 years. The hyperplasic scar fibroblasts were obtained generated from original culture in laboratory of surgical department.INTERVENTIONS: In the research, the experiment group and the control group were divided. In the experiment group, asiaticoside was applied on fibroblasts; in the control group, asiaticoside was not prescribed. The changes of every index were observed before and after medication.MAIN OUTCOME MEASURES: ① Effect of asiaticoside on expression of phosphorylated Smad2 and Smad7; ② Effect of asiaticoside on cell cycle and apoptosis.RESULTS: Asiaticoside inhibited scar fibroblasts entering M phrase from S phrase and reduced the content of phosphorylated Smad2 in fibroblasts, which did not present significant difference in two groups ( t = 1.53, P =0.08).The content of Smad7 in the cells was (50. 80 ± 22.40)% in the experiment group and (32.18 ± 17.84)% in the control group, which indicated significant difference ( t = 2. 17, P = 0. 024).CONCLUSION: Asiaticoside inhibits scar formation by Smad passage.

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.

ObjectiveTo explore the role of vascular endothelial growth factor(VEGF) in the growth and development of hypertrophic scar.MethodsThe burn wound samples of various stages were selected from transition of wound granulation tissue to scar and in long-persisting post-burn hypertrophic scar, and the concentrations of VEGF protein were detected using enzyme-linked immunosorbent essay (ELISA) method. ResultsThe tissue homogenate concentration of VEGF protein increases gradually from the wound granulation tissue to hypertrophic scar before it achieves summit concentration during 4 to 6 month. The concentration of VEGF degreases gradually after the maturation of hypertrophic scar. The high concentration of VEGF is synonymous with the large amount of capillary of the immature scar.ConclusionsThe abnormal expression of VEGF is related to the growth and development of hypertrophic scar and induces excessive and uncontrollable angiogenesis.

Objective To investigate the effect of 15d-PGJ2 on the expression of collagen type,CTGF and a-SMA in the hypertrophic scar in the rabbit ear,and the possibility of hypertrophic scar treated by 15d-PGJ2.Methods 18 New Zealand white rabbits were used to establish a hypertrophic scar model on the rabbit ear.The wounds were established as follows:2 cm × 3 cm wounds with total skin loses on the ventral side,2 wounds for each ear,totally 72 wounds.The wounds were randomly divided into the 15d-PGJ2 treatment group and NS control group.20μl 15d-PGJ2 or NS was injected into the ear scar once a day for 7 days.At 7,14 and 21 days after the injection,12 scars of each group were harvested.The expression of collagen type Ⅰ,CTGF and a-SMA was detected by immunohistochemical method.Results Excessive dermal scars on rabbit ears that were similar to human hypertrophic scar appeared in the two groups.Compared with the NS-treated scars group,the 15d-PGJ2-treated scars appeared to be smaller,softer,flatter and lighter in color.The expression of collagen type Ⅰ,CTGF and a-SMA in the 15d-PGJ2 group was significantly decreased as compared with that in the control group at different time points(P＜0.05).Conclusion 15d-PGJ2,the ligand of PPAR-r,can reduce the expression of collagen type Ⅰ,CTGF and a-SMA of hypertrophic scar in the rabbit ears and plays an important role in the prevention and treatment of hypertrophic scar.It may provide a new approach for the treatment of hypertrophic scar in clinical setting.

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