BACKGROUND: Compared with normal physiological flap, main advantage of venous skin flap is that it throws off the limitation of arterial vascular territory on donor site and recipient site of traditional axial skin flap. However, its survival rate is unstable. OBJECTIVE: To explore effects of basic fibroblast growth factor (bFGF)-poly(lactic-co-glycolic-acid)] (PLGA)-sustained release microspheres on the survival of rabbit venous flaps.DESIGN, TIME AND SETTING: A randomized controlled animal study was performed at the University of South China from May to October 2008.MATERIALS: A total of 24 healthy New Zealand rabbits were equally randomly assigned to bFGF-PLGA sustained release microsphere, blank microsphere and blank control groups.METHODS: The formulation of bFGF microspheres was optimized by orthogonal design. bFGF-PLGA microspheres were prepared by optimized method. Lateral abdominal wall skin flap was created in rabbits from 3 groups. Five days before operation, 28.85 g/L bFGF-PLGA microspheres 3 mL (containing bFGF 20 μg) was intradermally injected into rabbits from the bFGF-PLGA sustained release microsphere group. An equal volume of blank microsphere + bFGF was injected in the rabbits of the blank microsphere group. Rabbits from the blank control group were infused with the same volume of saline. MAIN OUTCOME MEASURES: Morphology and particle distribution of bFGF-PLGA microspheres, drug loading volume, encapsulation efficiency, in vitro drug release characteristics were measured. After seven days, the survival area of skin was determined. Rabbit skin samples received CD34~+ immunohistochemical staining to detect the expression of CD34~+ and average number of blood vessels. RESULTS: The bFGF microsphere prepared based on optimized formulation exhibited well-defined properties, with the even and uniform sphere in appearance, regular particles without adhesion, about 98% of particles with a size distribution between 12.50 to 43.49 μm, with a mean particle size of 26.93μm and size span of (0.611 ± 6.60). The drug loading volume and encapsulation efficiency of bFGF microsphere reached [(23.11 ±0.44 )x10~3]% and (86.51±0.83)%, respectively. In the burst release phase, the rate of in vitro drug release amounted to 27.78%, but rose to 81.56% accumulatively 30 days later. The in vitro drug release of bFGF microsphere corresponded with Higuichi equation (r= 0.997). The sustained-release microspheres, blank microspheres and normal saline group, the average survival of the flap and the average number of blood vessels were similar (P=0.597, P=0.336), but still significantly lower than the bFGF-PLGA sustained release microsphere group (P=0.000). Results of immunohistochemical staining revealed that bFGF-PLGA promoted blood supple between flap and surroundings, improved flap survival and abundant CD34~+ expression. CONCLUSION: bFGF microsphere with good morphology, high drug loading volume and encapsulation efficiency can be obtained using W/O/W multiple emulsion evaporation method. The bFGF microsphere can promote the survival of rabbit venous flaps through a long period due to sustained release of bFGF.

BACKGROUND:Compared with normal physiological flap,main advantage of venous skin flap is that it throws off the limitation of arterial vascular territory on donor site and recipient site of traditional axial skin flap.However,its survival rate is unstable.OBJECTIVE:To explore effects of basic fibroblast growth factor (bFGF)-[poly(lactic-co-glycolic-acid)] (PLGA)-sustained release microspheres on the survival of rabbit venous flaps.DESIGN,TIME AND SETTING:A randomized controlled animal study was performed at the University of South China from May to October 2008.MATERIALS:A total of 24 healthy New Zealand rabbits were equally randomly assigned to bFGF-PLGA sustained release microsphere,blank microsphere and blank control groups.METHODS:The formulation of bFGF microspheres was optimized by orthogonal design.bFGF-PLGA microspheres were prepared by optimized method.Lateral abdominal wall skin flap was created in rabbits from 3 groups.Five days before operation,28.85 g/L bFGF-PLGA microspheres 3 mL (containing bFGF 20 ?g) was intradermally injected into rabbits from the bFGF-PLGA sustained release microsphere group.An equal volume of blank microsphere + bFGF was injected in the rabbits of the blank microsphere group.Rabbits from the blank control group were infused with the same volume of saline.MAIN OUTCOME MEASURES:Morphology and particle distribution of bFGF-PLGA microspheres,drug loading volume,encapsulation efficiency,in vitro drug release characteristics were measured.After seven days,the survival area of skin was determined.Rabbit skin samples received CD34+ immunohistochemical staining to detect the expression of CD34+ and average number of blood vessels.RESULTS:The bFGF microsphere prepared based on optimized formulation exhibited well-defined properties,with the even and uniform sphere in appearance,regular particles without adhesion,about 98% of particles with a size distribution between 12.50 to 43.49 ?m,with a mean particle size of 26.93 ?m and size span of (0.611 ? 6.60).The drug loading volume and encapsulation efficiency of bFGF microsphere reached [(23.11?0.44)?10-3]% and (86.51?0.83)%,respectively.In the burst release phase,the rate of in vitro drug release amounted to 27.78%,but rose to 81.56% accumulatively 30 days later.The in vitro drug release of bFGF microsphere corresponded with Higuichi equation (r=0.997).The sustained-release microspheres,blank microspheres and normal saline group,the average survival of the flap and the average number of blood vessels were similar (P=0.597,P=0.336),but still significantly lower than the bFGF-PLGA sustained release microsphere group (P=0.000).Results of immunohistochemical staining revealed that bFGF-PLGA promoted blood supple between flap and surroundings,improved flap survival and abundant CD34+ expression.CONCLUSION:bFGF microsphere with good morphology,high drug loading volume and encapsulation efficiency can be obtained using W/O/W multiple emulsion evaporation method.The bFGF microsphere can promote the survival of rabbit venous flaps through a long period due to sustained release of bFGF.

BACKGROUND: The soft tissues would shrink with nasal framework collapse following surgical trauma, which cause aseptic inflammation, lead to parts of cartilage resorption. Accordingly, long-term saddle nose deformity usually accompanied by short nasal columella. Complications such as skin perforation or ulceration would appear if corrected the deformity using medical silicone rubber with large tension.OBJECTIVE: To explore the effectiveness of repairing post-traumatic saddle nose deformity with autologous costal cartilage transplantation.METHODS: A total of 21 cases with post-traumatic saddle nose deformity accompanied by short nasal columella were selected, including 6 males and 15 females, aged 16 45 years. All of the cases had trauma history and agreed with the treatment. The costal cartilage was obtained from the seventh rib and formed to babylon weeping willow leaf shape and columella nasi stent to repair post-traumatic saddle nose deformity. The "V-Y" progradation suture was used in the philtrum introcession and the botton of nasal columella to extend the nasal columella. The recovery of saddle nose deformity after transplantation, discharge of transplanted cartilage, as well as the incision scar status was observed.RESULTS AND CONCLUSION: The results were satisfactory and there were no complications after transplantation. All the cases were followed up from 6 months to 2 years. No case suffered costal cartilage grafts discharge or chondral deformation. The scar was little at the bottom of nasal columella. It is an ideal method for repairing post-traumatic saddle nose deformity using transplantation of autologous costal cartilage with "V-Y" progradation suture.

Objective:To evaluate the clinical effect of combination of autologous tissue reconstruction of tarsal plate with temporal flap on repair of full-thickness lower eyelid defect.Methods:Eleven patients (11 eyes) underwent hard palate mucosa or ear cartilage combined the emporal flap with the orbicularis oculi muscle to repair full-thickness defect ofpalpebra inferior.Of the 11 patients,6 had more than 75％ eyelid defect area,and 5 had more than 50％ eyelid defect area.Results:All 11 eyes closed completely,with no entropion or ectropion,and returned to normal basically.Postoperative follow-up was performed for 6 months to 5 years,3 years and 4 months on average.The function and form of eyelid remained stable.Infection,leakage or contracture was not found on reconstruction tarsus.Conclusion:Reconstruction of eyelid with autogenous hard palate mucosa or ear cartilage combined the emporal flap with the orbicularis oculi muscle is a simple,convenient and effective method.

[Abstract ] Objective High-mobility group box-1 (HMGB1) is abundantly released in the epileptogenic brain tissue , but few reports are seen about the effect of HMGB 1 on the expression of P-glycoprotein ( P-gp) in the vascular endothelial cells of the epi-leptogenic tissue .This study is to explore whether HMGB 1 can regulate P-gp expression in the brain microvascular endothelial cells of the mouse in vitro . Methods Immortalized brain microvascular endothelial bEnd .3 cells of the mouse were cultured in vitro and al-located to different concentration groups ( treated with culture medium containing 10 , 100 , 500 , and 1000 ng/mL HMGB1 for 8 hours), treatment duration groups (treated with culture medium containing 100 ng/mL HMGB1 for 4, 8, 16, 24, and 32 hours), and a control group ( treated with culture medium without HMGB 1 ) .The mRNA expression of P-gp-encoding gene-multidrug resistance gene 1a (mdr1a) was detected by real-time qPCR, and its protein expression determined by Western blot and immunocytochemistry . Results The results of qPCR manifested that the expressions of mdr 1a mRNA were 1.646 ±0.176, 1.777 ±0.135, 1.617 ±0.043, and 1.398 ±0.182 in the 10, 100, 500, and 1000 ng/mL HMGB1 groups, respectively, significantly higher than 1.030 ±0.284 in the control group (P<0.05), and so were those in the 4, 8, 16, 24 h, and 32 h groups (2.655 ±0.112, 2.168 ±0.212, 1.823 ± 0.232, 1.418 ±0.376, and 1.445 ±0.123) than in the control (1.010 ±0.164) (P <0.05).Western blot showed a significant increase in the P-gp protein expression in all the concentration groups (P<0.05) as well as in the 8 h and 16 h treatment duration groups as compared with the control group (P<0.05).Immunocytochemis-try also revealed a higher P-gp expression in the HMGB1-treated than in the control cells (P<0.01). Conclusion HMGB1 can upregu-late the expressions of mdr1a mRNA and P-gp protein in the brain microvascular endothelial cells of the mouse , which may associated with drug resistance of central nervous system diseases , especially that of epilepsy .

Objective To observe the roles of nuclear factor-κB (NF-κB) and hypoxia-inducible factor-1 o (HIF-1 α) in hippocampal neurodegeneration of status epilepticus (SE) rats, and explore whether HIF-1α activation is regulated by NF-κB.Methods A total of 110 male Sprague-Dawley rats were randomly divided into seven groups : (1) Control group treated with saline (control, n =15), (2) sham group implanted cannula into lateral ventricle and treated with saline (sham, n =15), (3) SE group treated with pilocarpine (SE, n =20), (4) NF-κB activity inhibitor pyrrolidine dithiocarbamate (PDTC) group treated only with PDTC (PDTC, n =15), (5) SE + PDTC group treated with pilocarpine plus PDTC (SE + PDTC, n =15), (6) SE + HIF-1o siRNA group implanted cannula into lateral ventricle and treated with pilocarpine plus HIF-1 α siRNA (SE + HIF-1α siRNA, n =15), (7) SE + control siRNA group implanted cannula into lateral ventricle and treated pilocarpine plus control siRNA (n =15).SE was induced by injecting lithium chloride and pilocarpine.The seizure of rats was observed.The protein expressions of NF-κB and HIF-1 α in hippocampus of rats were examined by Western blotting.The degenerating neurons in hippocampus were detected by Fluoro-Jade C (FJC) staining.Results Twenty-four hours after termination of SE, the nuclear protein expressions of NF-κB and HIF-1α in hippocampus of rats were increased in SE group (0.57 × 0.06, 0.47 ± 0.07) compared with those in control group (0.23 ± 0.03, 0.20 ± 0.03;P ＜0.05);and compared with SE group PDTC significantly decreased the nuclear protein expressions of NF-κB and HIF-1 α in SE + PDTC group (0.23 ± 0.03, 0.14 ± 0.03;P ＜ 0.05);in SE + PDTC group the numbers of FJC positive cells in CA1 area (28.33 ±5.03) were decreased compared with that in SE group (76.67 ± 13.32);HIF-1 o siRNA injected into lateral ventricle of rats significantly decreased the expression of HIF-1α in hippocampus (0.22 ±0.03) and the number of FJC positive cell in CA1 area (27.34 ±7.02) in SE + HIF-1α siRNA group compared with those in SE group (0.39 ±0.06, 76.67 ± 13.32;P ＜0.05).Conclusions These data suggest that SE can result in activation of NF-κB/HIF-1o pathway in brain.Inhibition of the pathway can attenuate hippocampal neurodegeneration caused by SE, which has the brain protective effect.

This paper adopted a series of related analysis methods to comprehensively analyze post-marketing clinical safety data of Shenmai injection from 4,220 cases of SRS and 32,358 cases of multicenter, prospective, registered hospital centralized monitoring in large data background, calculated ADR incidence rate was 0.93 per 1,000, main symptoms of ADR includes chest pain, chills, skin itching, palpitations, fever, nausea, dizziness, vomiting, flushing, numbness, allergic reaction, cyanosis, rash, low back pain, and "breath", "anaphylactoid reaction" and "flush" were the safety warning signals of Shenmai injection. Primary disease for chronic pulmonary heart disease, thyroid disease, and combined with cerebral vascular disease, prior to the injection and continuous use of alprostadil, cyclic adenosine monophosphate, combined with quinolones, penicillins were suspicious influence factors of ADR of Shenmai injection, these promot the clinical safety.
Drug Combinations ; Drugs, Chinese Herbal ; adverse effects ; Humans ; Injections ; Product Surveillance, Postmarketing ; Prospective Studies

Objective:To explore effects of trehalose as a cryopreserve agent on survival rate of fatty tissue after cryopreservation.Methods:The liposuction was used on the abdomen of adult female.After centrifugation and purification,adipose was randomized into the following three groups,the trehalose group,the fetal bovine serum (FBS)+ 10％DMSO group and the physiological saline group.The specimens were cryopreserved at-196 ℃ for 3 months and then the HE staining,glucose transfer method and CK method were used to detect the cell survival rate in each group.Results:The activity of adipose in the trehalose group and FBS+10％DMSO group adipose was higher than that in the physiological saline group (P＜0.05);while there was no significant difference between the trehalose group and FBS+10％DMSO group (P＞0.05).Conclusion:As cryoprotectant,trehalose could keep fat cell viability,and adipose tissue can be used for clinical transplantation after 3 months' freezing.

Purpose To identify the role of tyrosine kinase receptor Axl for anti-apoptosis which was induced by cisplatin (DDP) and methotrexate (MTX) chemotherapy and to analyze the relationship between P-Axl and apoptosis-related proteins in osteosarcoma.Methods Osteosarcoma cell lines MG63,143B and U2OS were used in apoptosis assays,Axl siRNA transfection,cytotoxicity assays,cell cycle analysis,etc.A total of 41 cases of osteosarcom patients were included for immunohistochemistry of EnVision two-step staining and clinico-pathological relative analysis.TUNEL assay was performed in ten cases for apoptosis detection.Results Among the osteosarcoma cell lines,Gas6/Axl could obviously protect tumor cells from apoptosis induced by DDP and MTX (P ＜ 0.05).Axl siRNA transfection enhanced cell apoptosis,whereas Gas6 prone to function upon previous knockdown by Axl siRNA.Among the 41 cases,the positive rate of P-Axl,BCL-2,and Bax was 85.4％,70.7％,and 36.6％,respectively.In contrast,the positive rate of them was 22.2％,11.1％,and 11.1％ in osteofibrous dysplasia,respectively.The expression levels of these apoptosisrelated factors were significantly higher in osteosarcoma than in osteofibrous dysplasia (P ＜ 0.05).Through clinico-pathological analysis,there were significant relationships between the survival status and BCL-2 or Bax expression (P ＜ 0.05).Pearson correlation analysis demonstrated that BCL-2 was positively correlated to P-Axl with statistical significance (r =0.842,P ＜ 0.0001).By Cox univariate analysis,BCL-2 or Bax was correlated with the patients' prognosis.TUNEL assay also demonstrated that P-Axl high expression inhibited apoptosis in osteosarcoma tissues.Conclusion Gas6/Axl protects osteosarcoma cells from the apoptosis induced by DDP and MTX chemotherapy and inhibits apoptosis in osteosarcoma tissue,possibly through the regulation of apoptosis-related protein BCL-2.

Objective:To understand the effect of a new tracheal tube fixation device in the "double fixation" of oral tracheal intubation for critically ill patients in adult emergency department.Methods:A retrospective analysis of patients with oral tracheal intubation admitted to our EICU from January 2016 to June 2017 was performed using traditional tape and inch band fixation as a control group; the experience of treatment from July 2017 to December 2018 for patients with oral tracheal intubation, the new double-fixation method of tracheal tube fixation device was used as the observation group. The degree of displacement, oral cleanliness, facial skin allergies and injuries, and comfort of the two groups of patients was compared.Results:The rate of tracheal displacement in the experimental group was mildly 11.2% (11/98), moderately 5.1% (5/98), and the control group was mildly 24.2% (15/62), moderately 14.5% (9/62) and severe 6.5% (4/62). The incidence of facial skin allergy and skin damage was 6.1% (6/98), 0, and the control group was 24.2% (15/62), 11.3% (7/62), the difference was statistically significant ( Z value was -4.021, χ2 value was 24.48, P<0.05). The comfort score of the test group was (1.05±1.01) points, which was significantly higher than (2.10±1.71) points of the control group ( t value was 4.920, P<0.01). Conclusions:The new type of tracheal tube fixation device "dual fixation" for critically ill patients with mechanical ventilation through oral endotracheal intubation is visually beautiful, and has good firmness and comfort. It can effectively reduce the occurrence of adverse events and has good application value.