Objective To explore the effect of epidermal growth factor (EGF) on tube formation of HUVEC induced by the secretion of angiogenesis factors of adipose-derived stem cells (ADSCs).Methods ADSCs were primarily cultured by enzyme digestion method.The flow cytomertry was performed to detect the expression of cell surface marker.ELISA was used to detect the expression of VEGF,HGF,and SDF-1 after given different doses of EGF.Tube formation assay was used to examine the effect of EGF on the tube formation induced by ADSCs.Results ADSCs were successfully isolated and cultured from human liposuction tissue and specific markers were expressed on ADSCs.EGF promoted the secretion of angiogenesis factors VEGF,HGF,and SDF-1,which were secreted by ADSCs.EGF pretreatment increased the ability of tube formation of HUVECs induced by ADSCs.Conclusions ADSCs induce the secretion of angiogenesis factors in vitro,and thus increase the ability of tube formation of HUVECs.EGF promotes the secretion ability of ADSCs,and the best concentration is 15 mg/L.

Objective To investigate the method of dermal-fat graft combined with secondary autologous fat transplantation in repairing severe facial depression and to evaluate the clinical effects.Methods Twelve cases of facial depression had been repaired by the transplantation of dermal-fat flap which was removed from the abdomen at the first stage.They were given fat granules injection 1 to 3 times postoperatively,and 3,6 and 3 patients were given fat granules injection three times,twice and once,respectively,with the interval period of 3 to 6 months.The result was based on comparison of the photos taken from preoperation and postoperation.Results All patients were healed primarily except one of which was formed hematoma after operation and scavenged thereafter.After 6 months to 2 years follow-up,all the patients had satisfactory facial contour.Conclusions Combined autologous fat granules with free dermal-fat graft to reconstruct severe facial depress is an easy,safe and effective technique and deserves to be recommended.

BACKGROUND:Early vascularization is crucial for wound healing. A high-porosity, macrovoid alogeneic skin leads to the rapid vascularization and celular infiltration.
OBJECTIVE: To obtain a new alogeneic skin product with high porosity, good cel permeability and good histocompatibility using an improved preparation method of human acelular dermal matrix.
METHODS: Cel components of healthy human skins were removed by the improved method and the traditional method, respectively. The improved method was to remove the subcutaneous fat, eliminate the epidermis (1 mol/L NaCl solution at 37℃ for 24 hours) folowed by shaking processing (2% NaOH at 45℃ for 4 hours), and then, the solution was neutralized with PBS rinsing, dried and stored at 4℃ for standby. We detected the porosity and degradation time in vitroof the acelular dermal matrices prepared by two methods and the cytotoxicity of the material infiltration liquid on the adipose-derived stem cels. Hematoxylin-eosin staining was used for the detection of the cel residual, the integrity of colagen and cel biocompatibility. Scanning electron microscopy was used to detect the pore size.
RESULTS AND CONCLUSION: Both the two methods could completely remove the cel components, and maintain the integrity of the colagen scaffold. The porosity of acelular dermal matrix with the improved method was (93.22±0.99)%, which was significantly higher than that with the traditional method [(74.28±2.06)%;P < 0.001]. However, there was no significant difference in in vitrodegradation time between the two kinds of acelular dermal matrices(P > 0.05). No obvious cytotoxicity of the acelular dermal matrix prepared with the improved method was detected. At 3-7 days of co-culture, the adipose-derived stem cels cultured on the acelular dermal matrix prepared with the improved method could penetrate the basement membrane to the deep dermis, while there was no obvious cel invasion and growth in the deep dermis prepared by the traditional method. Compared with the traditional method, the improved method is more suitable for cel infiltration and growth with higher porosity and larger pore size.

Objective To explore the effects of recombinant plasmids of pGPU6/GFP/NeoshRNA-CTGF (shRNA-CTGF) on the type Ⅰ collagen (COL-Ⅰ) protein expression in keloid,through RNA interference on connective tissue growth factor (CTGF) in vivo and in vitro.Methods Recombinant plasmids were designed and constructed by specific shRNA-CTGF; After transfeced human keloid fibroblast with shRNA-CTGF in vitro,RT-PCR was used to detect the CTGF mRNA level,and Western blot to detect the secretion of COL-Ⅰ.After transfected the keloid of nude mice with shRNA-CTGF in vivo,RT-PCR was used to detect the CTGF and COL-Ⅰ mRNA level,and Western blot was used to detect the protein expression of COL-Ⅰ.Results Recombinant plasmids of CTGF were successfully constructed,and the CTGF gene expression was significantly decreased in vivo and in vitro by 86.8％ and 54.1 ％,respectively; Down-regulation of CTGF in vitro significantly inhibited the mRNA and protein level of COL-Ⅰ by 76.8％ and 65.6％,respectively; Down-regulation of CTGF in vivo significantly reduced the COL-Ⅰ mRNA and protein level by 52.7％ and 48.0％,respectively.Conclusions CTGF gene expression is successfully down-regulated by the recombinant plasmid of shRNA-CTGF in vivo and in vitro.shRNA-CTGF significantly reduces the COL-Ⅰ protein level in keloid.It implies that CTGF gene is a potential target in the therapy of pathological scar.

Objective To evaluate the clinical efficacy of “Shamrock” ultrasound images?guided lumbar sympathetic ganglion blockade ( LSGB) . Methods Sixty patients of both sexes, aged 18-60 yr, weighing 50-70 kg, of American Society of Anesthesiologists physical statusⅠorⅡ, undergoing unilater?al LSGB, were divided into groupⅠ ( n=30) and group Ⅱ ( n=30) using a random number table. In group Ⅰ, unilateral LSGB was performed at the L2 level under ultrasound guidance with paramedian trans?verse scanning. In groupⅡ, unilateral LSGB was performed at the L2 level under ultrasound guidance with“Shamrock” ultrasound images. After final needle position was confirmed, 2% lidocaine 6?0 ml was ad?ministered in each patient. At 20 min before and after LSGB, the visual analogue scale scores and skin temperature of the big toe of the affected foot were recorded, and the successful blockade and visibility of important paravertebral structures on ultrasound images were recorded during puncture. Results The visu?al analogue scale scores were significantly lower, and the skin temperature on the affected side was signifi?cantly higher after LSGB than before LSGB in both groups ( P<0?05) . The important paravertebral struc?tures such as erector spinae, quadratus lumborum, psoas major, transverse process of L2 vertebrae, and the curved edge of L2 vertebrae were visible in both groups. The visibility rate of the inferior vena cava or ab?dominal aorta on ultrasound images and the success rate of blockade were significantly lower in group Ⅰthan in group Ⅱ (P<0?01). Conclusion Compared with paramedian transverse scanning, LSGB has some advantages such as real?time monitoring, higher success rate of blockade, better efficacy and avoiding damage to great vessels when performed under “Shamrock” ultrasound image guidance.

Objective:To investigate the prevalence of chronic obstructive pulmonary disease (COPD) in the adults aged 40 years old and over in Changchun urban area,and to clarify the important significance of early application of pulmonary function examination.Methods:With the help of community workers,10 communities which around Second Hospital of Jilin University were selected and the participants aged 40 years old and over were randomly collected.COPD questionnaire and pulmonary function examination were conducted in all subjects.The patients with COPD were diagnosed according to the criteria established by the Global Initiative for COPD (GOLD).The patients with COPD were divided into the diagnostic group and undiagnostic group,then the patients with COPD who had smoking history were divided into smoking cessation group and smoking group.The pulmonary function of each groups were compared.Results:A total of 1 392 participants aged 40 years old and over were recruited in this survey,including 568 males and 824 females,and their average age was (59.59±6.60) years.Totally 176 subjects were diagnosed as COPD,and the prevalence was 12.64％.Furthermore,160 of them were newly diagnosed,which showed the undiagnostic rate of COPD was 90.91 ％.Among the 176 patients with COPD,16 had been confirmed before,which meant only 9.09％ patients got the accurate diagnose before.The analysis of high-risk factor (smoking) showed that the population had an increased prevalence of COPD (20.71％) compared with non-smokers (8.8％) (P＜ 0.05).The correlational analysis between smoking index and the pulmonary function of the patients with COPD showed that all of the pulmonary function indexes except forced vital capacity (FVC％) and maximum midexpiratory flow (MMEF％) in smoking cessation group were descended along with the increase of smoking index (P＜0.05).All of the pulmonary function indexes except FVC％ in smoking group were descended along with the increase of smoking index (P ＜ 0.05).Conclusion:The high prevalence and underdiagnostic rate of COPD are found in the residents aged 40 years old and over in Changchun urban area.More patients with COPD should be screened by performing pulmonary function examination to treat them at an early stage.

Objective To explore the operating methods and its related questions of auricular reconstruction with totally expanded skin in combination with laser hair removal for the treatment of adolescent microtia.Methods From Jan.2013 to Dec.2016,30 adolescent microtia patients were treated with totally expanded skin.At the first stage,the 100 ml kidney-shaped expander was implanted under the skin of mastoid.After expanding capacity of 80 ml,the hair on the expanded skin was depilated once a month with reference to the healthy ear;at the second stage,after expanding capacity of 150 ml,the expander was taken out and the fiber capsule was removed;the tautologous rib cartilage was harvested and the scaffolds were sculptured;the cartilage was implanted and the expanded skin flap was used to cover the frontal surface and back surface of the scaffold;at the third stage,the earlobe transposition,conchal excavation and tragus construction were performed at the same time.Results All the patients were followed up for 3 to 24 months;the results showed 1 case of leakage of expander,4 cases of hematoma,2 case of expanded skin burst,and the complications were treated correctly,all patients were satisfied with the appearance;the color,texture,location,size;and height of ear cranial angle were matched with health ear;there was no obvious scar and auricle subunit structure was clear.Conclusions The laser in combination with the large capacity tissue expander in auricular reconstruction is simple,less trauma and less scarring.

Objective To evaluate the changes in the expression of neuroligin1 in excitatory postsynaptic membrane of the spinal dorsal horn in a rat model of inc isional pain.Methods Forty-eight pathogen-free healthy adult male Sprague-Dawley rats,aged 6-8 weeks,weighing 280-320 g,were divided into control group (group C,n =12) and incisional pain group (group Ⅰ,n=36) using a random number table.A1 cm long incision was made in the plantar surface of the right hindpaw in group Ⅰ.Cumulative pain score (CPS) was assessed and mechanical paw withdrawal threshold to von Frey stimuli was measured at 3 hand 1 and 3 days after operation (T1,2,3).The animals were then sacrificed and their lumbar segments (L3-6) of the spinal cord were removed for detection of the expression of neuroligin1,postsynaptic density-95 protein (PSD-95),glutamate receptor 1 (GluR1) and GluR2 in the postsynaptic membrane of spinal dorsal horn (by Western blot) and co-expression of neuroligin1 with PSD-95 in spinal dorsal horn (by co-immuno-precipitation).Results Compared with group C,CPS was significantly increased and mechanical paw withdrawal threshold was decreased at T1-3,and the expression of neuroligin1 and GluR1 in the postsynaptic membrane of spinal dorsal horn at T1,2 and co-expression of neuroligin1 with PSD-95 at T1 were up-regulated in group Ⅰ (P＜0.01 or 0.05).Conclusion The development and maintenance of incisional pain is related to the signaling pathway regulated by neuroligin1 in excitatory postsynaptic membrane of the spinal dorsal horn of rats.

Objective To study the cell morphology and differentiation efficiency when rabbit bone marrow mesenchymal stem cells (BMSCs) were induced osteogenic differentiation as culturing by autologous platelet-rich plasma (PRP) instead of serum,and to explore a new method of inducing BMSCs osteogenic differentiation.Methods The PRP was prepared by arterial blood of rabbit.Punctured and The bone marrow was sampled from rabbit's iliac bone,and BMSCs were collected,which divided into PRP group,fetal calf serum (FBS) group and serum-free control group,and cultured in 10％ autologous PRP,10％ FBS and serum-free respectively,combined with DMEM-F12 medium.The second generation cells were divided into experimental and control groups.The experimental groups' medium was added dexamethasone,β-sodium glycerophosphate and ascorbic acid,and the control groups went on.The cell morphological difference of each group was Observed between anterior and after inducing differentiation,and compared between each group.Results BMSCs of PRP and FBS groups grew quickly,presented like fusiform form before induction,and increasd in volume,became a triangle,polygonal and round form gradually after osteogenic induction.Cells of PRP and FBS groups aggregated spontaneously and multilayered,and formed calcium nodules and bone-like structure after induced 7 days averagely,which could be stained red by alizarin red S;cells of serum-free groups were induced 14 days averagely,only three samples showed osteogenesis performance.Cells of PRP and FBS groups differentiation efficiency was superior to serum-free groups when inducd 20 days,the difference was statistically significant (P＜0.05),and the difference between efficiency of PRP and FBS groups was not significant (P＞0.05).Conclusions Autologous PRP could be used to proliferate and induce osteogenic differentiation of BMSCs instead of serum.

Objective To observe the clinical efficacy of propranolol and 595 nm pulsed dye la ser (PDL) in treatment of infantile hemangioma.Methods 26 infants admitted to our hospital from January 2013 to January 2015 with hemangioma underwent oral propranolol 2 mg/(kg · d) treatment after excluding of taboos.The daily doses were divided equally to two parts,taken on the time of 8:00 and 20:00,when the electrocardiograph and pulse oxygen were monitored and recorded persistently.The patients were discharged from the hospital when it was stable,with review of blood routine examination,fasting blood glucose,liver and kidney function,and the change of size,character and color of hemangioma were recorded,and taken photos every two weeks after discharge.The 595 nm PDL was used to treat the hemangioma faded incompletely when the propranolol was terminated.Results The tension and color of all hemangioma decreased in varying degrees in taking propranolol for 72 hours,and evaluated the efficacy as recovery completely 19 cases;signifivantly effective in 3 cases and partial efficacy in 4 cases;the latter 7 cases were further treated with 595 nm PDL.Followed-up for 6-12 months showed that efficacy of recovery reached 100％.10 cases showed heart rate was mild reversibly slow,with no special treatment.5 cases had diarrhea,and healed with symptomatic treatment.No adverse reactions like liver and kidney dysfunction and so on were found.Conclusions Propranolol and 595 nm PDL can effectively treat infantile hemangioma,and thus it can be used as the recommended treatment of infantile hemangioma.