10.3969/j.issn.1000-8179.2013.12.016

Objective To observe the efficacy of pulsed dual-wavelength 595 and 1064 nm laser in treating ulcerated infantile hemangioma.Methods Lesions from 40 patients were treated with the laser.The distribution of lesion sites was in buttuck and perianal area (13 patients),vulvar lips (7),groin and inside thigh (5),scrotum (4),neck (3),scalp (2),lip (2),trunk and limbs (4).All lesions including ulcer and non-ulcer were conducted with 595 nm plused dye leaser at the first therapy.Later,595 nm pulsed dye leaser was continued or performed with Group 1 or Group 2 parameter of 595 and 1064 nm multiplex mode until the treatment was finished according to the thickness of the lesion.The frequency of treatment was one to six times.The interval of the treatment was four weeks.Results The ulcer from 39 lesions healed gradually in 1 to 2 weeks (average 9.5 days) after the first treatment.These ulcer lesions were healed four weeks after the first time of therapy.The recovery rate of ulcerated lesion was 97.5 ％.For the whole lesion,after 1 to 6 treatments,13 lesions showed excellent result,19 lesions showed good result,and the effective rate was 80 ％.Pain relieved 2 to 7days (average 3.5 days) after the first laser therapy in 35 patients who accompanied with pain.Pain score decreased from 1.875 to 0.125 before and after treatment,with significant difference(P＜0.05).The tolerance of the treatment was good and no side effect was observed.Conclusions 595and 1064 nm dual-wavelength laser is a safe and effective tool for treating ulcerated infantile hemangioma.

Objective To explore the influence of injury severity on transplantation of embryonic neural stem cells (NSCs) after traumatic brain injury (TBI).Methods The NSCs were isolated from the hippocampus of fetal rats aged at from 12-14 days.The cells were cultured and proliferated in the serum-free medium and identified in vitro.The animals received transplants in the bilateral hippocampal areas at day 3 following mild or moderate TBI separately.Conventional histology,TUNEL and immunohistology were examined to detect BrdU,NSE,GFAP,GalC,NGF and BDNF at day 14 post-implantation.Results BrdU-labeled positive cells in the bilateral hippocampus in the mild TBI group were more than those in the moderate TBI group at day 14 post-implantation.Significant differentiation of the astrocytes recognized as GFAP positive cells in the bilateral hippocampus was found at day 14 post-implantation.The expression of NGF and BDNF proteins was increased following TBI,the most evident in the mild TBI group.Conclusion The influence of injury severity on transplantation may be associated with the change of the microenvironment after TBI.

Objective To study the mechanism of interaction between neuronal nitric oxide syn-thase (nNOS) and inducible nitric oxide synthase (iNOS) following traumatic brain injury (TBI) in rats. Methods A total of 250 male Wistar rats were randomly divided into five groups, ie, sham oper-ation group, trauma group, 7-nitroindazole (7-NI) treatment group, aminoguanidine (AG) treatment group and combined AG and 7-NI treatment group. Severe closed TBI was made by using Marmarou meth-od. Protein expressions of nNOS and iNOS in hippocampus CAI were detected by means of immunohisto-chemical staining at 1,3, 6, 12 hours and at days 1,3, 7 and 14 after TBI. Results The expression of nNOS reached a peak at 6 hour after injury in all groups, with no statistical difference between groups (P > 0. 05), when there was no statistical difference between 7-NI treatment group and trauma group (P > 0. 05) but statistical difference in AG treatment group and combined AG and 7-NI treatment group compared with trauma group at 12 hours after TBI (P <0.05). The expression of iNOS reached maximal level at day 3 after TBI, with lower level in 7-NI group, AG treatment group and combined AG and 7-NI treatment group compared with trauma group (P < 0.05). Conclusions After TBI, nNOS interacts with iNOS by means of the feedback of nitric oxide. The enhanced expression of nNOS is initial factor for increase of iNOS expression, which can down regulate the expression of iNOS.

Objective To optimize human acellular dermal matrix(ADM) and evaluate its biological characters. Methods Human skin was treated with hypertonic saline followed by NaOH maceration(group A), hypertonic saline followed by sodium dodecyl sulfate (SDS) detergent(group B) or Dispase Ⅱ followed by Triton X-100(group C), the resulting ADM were sectioned, and then were stained by special immunohistochemistry method. The cytotoxicity of them were evaluated by methyl thiazolyl tetrazolium (MTT) colorimetry and then cell compatibility was analyzed by cell culture;The optimized ADM resulted was choosen for use. Fibrablasts(FBs)were transfected with adenovirus vector encoding green fluorescent protein gene(Ad-GFP)and the growth of them on the optimized ADM was observed by fluorescent microscopy. Results Collagen and elastic fibers can still be observed in three kinds of ADM. The cells in dermis can be disintegrated both in group A and C, but not in group B. The cytotoxicity scores of the ADM prepared in group A and B were grade 0 or grade 1, while that of group C was more than grade 1.The ADM prepared by NaCl-NaOH maceration had good biocompatibility. There was statistical difference in adhering number of NIH3T3 cells in group A and B. NIH3T3 cells grew well in group A and the resulted ADM was optimized. FBs transfected with Ad-GFP grew well in the optimized ADM. Conclusion The ADM prepared by NaCl-NaOH maceration was a good tissue engineering biomaterial with a little cytotoxicity and rich in resouce.

Objective To study the mechanism of dermal fibroblasts as a feeder layer to support the growth of human keratinocytes. Methods Human dermis fibroblasts were isolated and cultured and then treated with mitomycin-C. The expression of type Ⅰand type Ⅲ precollagen mRNA and relevant protein in feeder layer were examined by RT-PCR and Immunohistochemistry. KCs were cultured both on FB and NIH3T3 feed layer as control, the adhering numbers and the time of fusion were recorded. Results RT-PCR showed an increase of type Ⅰprecollagen mRNA in FB feeder layer as compared with that of normal fibroblasts (P

Objective To explore the mechanism of neurotoxic effects of neuron nitric oxide synthase(nNOS)and inducible nitric oxide synthase(iNOS),and the therapeutic effects of 7-nitroindazole(7-NI)and aminoguanidine(AG),in traumatic brain injury(TBI)rats.Methods Two hundred and fifty adult male Wistar rats were randomly divided into 5 groups:sham operation group,traumatic group,AG group,7-NI group and AG+7-NI group.Animals in each group were divided into 8 subgroups according to the time after trauma(1,3,6,12 hours and 1,3,7,14 days).Severe diffused brain injury model was made with Marmarou method.Expressions of nNOS and iNOS in hippocampus CA1 region were determined by immunohistochemistry,nerve cells apoptosis in hippocampus CA1 region was observed by TUNEL methods,and the relationship between apoptosis and NOS was observed by double staining.Results In trauma group,the expression of nNOS in hippocampus CA1 region peaked at 6h post-trauma,the expression of iNOS and apoptosis of nerve cells in hippocampus CA1 region both peaked at 3d post-trauma,while the apoptosis was alleviated in AG group,7-NI group and AG+7-NI group.The number of both TUNEL staining and nNOS immunostaining positive cells increased at 6h post-trauma in trauma group,significantly higher than that in 7-NI group.The number of both TUNEL staining and iNOS immunostaining positive cells increased at 3d post-trauma in trauma group,significantly higher than that in AG group.Conclusions The over-expression of nNOS and iNOS has toxic effects to neural tissues of brain,serves as one of the factors inducing nerve cell apoptosis in hippocampus CA1 region.7-NI and AG can inhibit the expression of nNOS and iNOS,reduce the nerve cell apoptosis,and play an important role in neuroprotective effect.

Objective To study the relationship between energy metabolic changes tested by phosphorus-31 magnetic resonance(MR)spectroscopy(31P MRS)and the liver damage score(LDS)in rabbit models and investigate the diagnostic value of 31P MRS in acute hepatic injury.Methods A total of 30 rabbits were received different radiation dose(ranging from 5 Gy to 20 Gy)to establish acute hepatic injury models.31P MRS was than carried out 24 hrs after radiation.The rabbits were divided into mild(LDS≤3 U),moderate(LDS 3-6 U)and severe(LDS＞6 U)hepatie injury groups.Ten healthy rabbits were served as controls.MR examination was performed on a 1.5 T imager using a 1H/31P surface coil by the 2D chemical shift imaging technique.The relative quantification of phosphomonoesters(PME),phosphodiesters (PDE), inorganic phosphate (Pi) and adenosine severe hepatic injury groups were 1.83±0.33,1.55±0.24,1.27±0.07 and 0.98±0.18,respectively,with significant differences(P＜0.05).The ATP level was progressively decreased with the increase of severity Of hepatic injury.The relative quantification of PME and Pi were decreased in severe hepatic injury group compared to control group(P＜0.05).There was no significant difference higher in moderate(1.94±0.50)and severe(1.96±0.72)hepatic injury groups compared to control group(1.43±0.31)and mild hepatic injury group(1.40+0.38)(P＜0.05).No significant difference was found in ratio of relative quantification of other phosphorus metabolites.Conclusions 31P MRS is a useful method in evaluating acute hepatic injury.The relative quantification of hepatic ATP level,which can reflect the severity of acute hepatic injury,is correlated with LDS.

Objective To determine the clinical value of pulsed dual-wavelength 595 and 1064 nm laser in treating superficial infantile hemangioma. Methods The treatments were conducted in 260 patients with 270 lesions. The location of the lesions were 41 cases in scalp, 88 in face and neck, 79 in trunk and 62 in arms and legs. All lesions were treated several times with 595 nm (Group 1) and/or 1064 nm (Group 4) multiplex mode until the treatment was finished. The frequency of treatment was one to six times. The interval of the treatment was four weeks. Results The total effective rate was 95.56 %, with atrophic scar (8.15 %) but without severe ulcer. No significance was found as compared Group 1 parameter with Group 4 in formation of atrophic scars (x2 = 1. 870, P＞0.05). Significant difference in the incidence of atrophic scars was found among different location of lesions (x2 = 15. 743, P＜0. 01) : lesion in face had the highest probability. No obvious difference was found between different location of lesion and the frequence of treatment (x2 = 21.164, P＞0.05), and so did with the efficacy (x2 = 11. 597, P＞0.05). The size and the thickness of lesion had significant difference in the time of treatment (x2 = 58. 171, P＜0. 01 and x2 = 11. 583, P＜0.05, respectively).Conclusions 595nm/1064 nm dual-wavelength laser is a safe and effective tool for treating superficial infantile hemangioma. Choice of treatment parameter in the mode depends on the features of vascular lesions.

Objective To investigate the effects of weightlessness on the apoptosis of hepatocytes in rats.Methods Eighty-four adult male Wistar rats were randomly divided into weightlessness group and control group according to the random number table.Tail-suspension was used to simulate the weightlessness model.Six rats were sacrificed in each group at the end of day 1-7.Hepatocyte apoptosis and p53 expression were detected by TUNEL assay and immunohistochemistry PV-6001,respectively.All data were analyzed by variance analysis.Results An increase of apoptosis index and up-regulated expression of p53 were detected in weightlessness group.The level of apoptotic index Was significantly higher in weightlessness group than in control group from day 1 to day 5(F=77.608,P<0.05),and the apoptotic indexes of the 2 groups were close at day 6 and day 7.The p53 proteins with positive expression were mainly detected in the nucleus.The level of positive expression of p53 in liver tissue in weightlessness group was higher than that in control group at the early stage(F=113.063,P<0.05).Few hepatocytes with positive expression were detected in control group and weightlessness group at a later stage.Conclusion There is a close relationship between hepatocyte apoptosis,changes of P53 expression and stress response and tolerance to weightlessness.