ObjectiveTo investigate the influence of connective tissue growth factor (CTGF) onthe collagen metabolism in human keloid fibroblasts with RNA interference (RNAi).Methods Human keloid fibroblasts (KFB) in vitro were transfected by 3 pairs of specific small interfering RNA (siRNA) CTGF plasmid synthesized for human CTGF,respectively.Reverse transcriptase-polymerase chain reaction (RT-PCR) contributed to the screening of the best siRNA in interfering of CTGF expression in human keloid fibroblasts to construct the plasmids,with the application of RNAi,to test the changes of expression level and collagen content of CTGF in transfected keloid fibroblasts through RT-PCR and Western blotting compared to its control groups.ResultsThe 3rd pair (C3) siRNA- CTGF expression of genes and proteins was remarkably inhibited after being interfered with human keloid fibroblasts,with inhibitory rates of 86.8 ％ and 65.6 ％.ConclusionsKeloid fibroblasts transfected by plasmid siRNA-CTGF effectively inhibite the expression of CTGF and deposition of collagen,and CTGF promotes the collagen synthesis in keloid development.

Objective To investigate the effect of butylphthalide on the memory,movement and ERP (event-related potentials) in patients with cerebral infarction cognitive impairment.Methods A total of 134 patients with cerebral infarction cognitive impairment were randomly divided into treatment group (67 patients) and control group (67 patients).All patients were given conventional cerebral infarction treatment as well as cognitive improvement intervention.Oral administration of butylphthalide was added onto the patients in the treatment group.Curative effects were observed after 30 days′ continued treatment.The cognitive function scores,memory scores,movement scores,ERP test and adverse reaction were compared between the two groups before and after treatment.Results After 30 days′ treatment,two groups of the montreal cognitive assessment scale (MoCA) scores were 27.2±4.9 and 25.1±4.1 respectively;the total scores of memory were 5.5±0.5 and 4.9±0.5 respectively;the scores of motor assessment scale (Fugl-meyer) were 85.6±6.2 and 74.2±6.1 respectively and P3 amplitudes were (9.5±0.9)μV and (8.1±0.9)μV respectively.All the indexes mentioned above in treatment group were significantly increased compared with those of before treatment(P<0.05).The results of N1,N2,P2 and P3 in the latency detection were significantly decreased compared with those of before treatment(P<0.05).And the results of P2 and P3 in the latency detection of the treatment group were shorter than those of the control group(P<0.05).After treatment,no adverse reaction was observed in both groups.Conclusion Based on conventional cerebral infraction treatment,oral administration of butyraldehyde effectively improve the therapeutic effect.

Aim To study the proliferative effect of resveratrol pretreatment on oxygen-glucose deprivation/reoxygenation ( OGD/R ) injury of rat cortical neural stem cells ( NSCs ) in vitro. Methods Isolation and purification of NSCs in neonatal Sprague-Dawley( SD) rats were conducted by suspended cultivation. The third passage NSCs of adherent culture was cultured under oxygen and glucose deprivation for 150 min and reoxygenation for 24 h. The experimental subjects were divided into normal, control, ethanol and resveratrol pretreatment groups. Immunofluorescence was used to identify NSCs. Cell viability was detected with CCK-8 assay. Flow cytometry cell cycle and BrdU assay were used to measure cell proliferation. Results Cells both in suspended and adherent cultivation highly expressed neuroepithelial stem cell protein ( nestin ) . Compared with the control group, NSCs viabilities and prolifera-tion in resveratrol groups (1, 5, 20 μmol·L-1 ) were significantly heightened, and highest in the 5 μmol · L-1 resveratrol group ( P<0. 05 ) . Conclusion Res-veratrol pretreatment can reduce injury and promote proliferation of NSCs after oxygen-glucose deprivation /reoxygenation.

Objective To investigate the specific silencing of connective tissue growth factor (CTGF) in a nude mouse keloid model,using RNA interference (RNAi) technique,and to provide the basis for gene therapy of keloid.Methods The nude mouse keloid model was established,and then transfected in vivo with well-amplifiating plasmid.The mRNA expression levels of CTGF mRNA and type Ⅰ collagen mRNA were detected with reverse transcription-polymerase chain reaction (RT-PCR).The distribution and protein expression levels of CTGF and type Ⅰ collagen were determined quantitatively using immunohistochemistry.Results The expression of CTGF at mRNA and protein levels was decreased in the experiment group,and the expression of type Ⅰ collagen at mRNA and protein levels was also decreased after transfection,as compared with negative control group and blank group,with significant difference between groups (P＜0.05).Moreover,the expression of type Ⅰ collagen and CTGF was positively correlated (r=0.979).Conclusions Keloid type Ⅰ collagen can be decreased through in vivo inhibiting CTGF expression.The transfection of CTGF gene in vivo may have effects on type Ⅰ collagen generation,and thus inhibit the keloid growth.

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 study the effect of the lentivirus encoding acidic fibroblast growth factor transfecting human adipose-derived stem cells (ADSCs) on the cell cycle and proliferation of ADSCs.Methods ADSCs were isolated and extracted by enzymatic digestion from the liposuction aspirate.ADSCs were cultured,identified and osteogenic induced reagent was used to induce differentiation of ADSCs towards bone cells.To obtain lentivirus encoding FGF-1,the plasmid PWPXLd FGF-1 was co-transfected with plasmid psPAX2,pMD2.G in 293T cells.ADSCs were infected with lentivirus encoding FGF-1.Expression of green fluorescent protein (GFP) in infected FGF-1 was observed by fluorescence microscope and expression of FGF-1 in ADSCs was verified by Western blot analysis.Flow cytometry was used to detect the cell cycle of ADSCs infected with lentivirus encoding FGF-1.EDU assay was performed to examine cell viability.Results Lentivirus encoding FGF-1 was obtained.After ADSCs being infected green fluorescence was found in about 70％ ADSCs,and overexpression of FGF-1 protein was detected in infceted cells by Western blot analysis.The percentage of G2/M phase cells was significantly increased compared with the control group,and the proliferation of ADSCs infected with lentivirus encoding FGF-1 was promoted as compared with the control group.Conclusions FGF-1 can enhance G2/M phase of ADSCs and promote the proliferation of ADSCs.

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 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 discuss the application of deep hypothermic circulatory arrest in surgical treatment of complex thoracoabdominal aortic aneurysms and its near-midterm effect.Methods The clinical data of 34 cases of thoracoabdominal aortic aneurysm in the center from August 2009 to June 2018 were analyzed retrospectively.All the patients underwent surgery under deep hypothermic circulatory arrest.There were 23 males and 11 females; aged 23 -67 years, mean(42.26 ±10.96) years old; Crawford type Ⅰ in 12 cases and Crawford type Ⅱ in 22 cases; aneurysms with a maximum diameter of 50 -120 mm, mean(65.26 ±16.09) mm;Marfan syndrome 15 cases, atherosclerosis 14 cases, aortic coarctation in 5 cases;22 cases of hypertension;28 cases of first aortic surgery, 6 cases of re-aortic surgery.Surgical transthoracic and abdominal incision, ext-racapsular approach, femoral artery and inferior vena cava intubation, deep hypothermic circulatory arrest technique to complete proximal anastomosis, arterial tube reconstruction of intercostal artery, abdominal organ blood supply artery and four The bifur-cated vessels were anastomosed, and the bifurcated vessels were anastomosed with the "Y"type artificial blood vessel trunk. The bilateral radial arteries were end-to-end anastomosis in the 10 mm artificial blood vessels of the "Y"type artificial blood vessels.Results There were no complications of cranial nerve system in the whole group , deep hypothermic circulatory arrest (17.68 ±4.88) min, ventilator assist time(34.88 ±16.04) hours, postoperative renal failure in 5 cases, after CRRT treat-ment After recovery, 1 case of paraplegia after operation, muscle strength recovered after cerebrospinal fluid drainage and de-compression, and 1 case died in the whole group, and died of multiple organ failure.The patients were followed up for 3 months to 5 years, and the results were satisfactory.The survivors did not die.The survivors did not die.However, 5 patients underwent thoracic aortic replacement under deep hypothermic circulatory arrest for the first time , and 4 patients underwent reo-peration because of distal vasodilation.The reconstructed intercostal artery occlusion occurred in 4 patients, but no paraplegia occurred.Conclusion When cross clamping the aorta is not feasible,it is safe to perform proximal anastomosis with deep hy-pothermic circulatory arrest.

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.