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 construct the recombinant plasmid carrying shRNA to ATX and analyze the nucle- ic acid sequence for further searching new gene therapy method of tumor.Methods Two DNA sequences containing short hairpin structure were designed and synthesized.The complement form was obtained by annealing and inserted into vector Psilcncer2.1-U6 neo,and the recombinant plasmid was transformed into DH5a strain.Finally the plasmid identified by restriction enzyme was used for sequence analysis.Results The recombinant Psileneer2.1-U6 neo car- rying shRNA to ATX had been constructed and the aim sequence had been obtained.Conclusion The construction of the recombinant plasmid carrying shRNA to ATX lays the basis for the study of its inhibitive effect on tumor.

Objective To analyze the operative effect of fracture of ulna coronoid process and evalu- ate the repairing effect of reconstruction of combined injury of ligamental structure or radius head.Methods During 2001 to 2004,16 cases(14 males and 2 females)of fractures of ulna coronoid process,aged 18-54 years(mean 36.6 years),were treated operatively in our department.Routine medial approach on the elbow or anterior trance-joint approach was used.Once reduction was achieved,screws or Kirschner wires were used to fix the fracture and the capsule was repaired at the same time.If there was combined fracture of the radius head or rupture of the collateral ligament,simultaneous repair or resection was applied.Functional exercise was requested in all patients.Results The follow-up was 8 to 24 months.Fracture union needed 4 to 6 weeks. Average Mayo elbow score was 80.Conclusion Ulna coronoid process fractures need active therapy. The radius head should be reserved as complete as possible during treatment.It is necessary to work out a standard management plan,and a notice should be taken to ligamental structure examination.

To analyze the regularity in combined medication with Xiyanping injection (Xiyanping for short) in the real world by as- sociation rules. Totally 5 822 patients using Xiyanping injection was collected from the 18 Class III Grade I hospitals nationwide to study the combined medication information of the patient with lung infection and make the analysis by using association rules and Apriori. According to the results, major drugs combined with Xiyanping in treatment of lung infection included compound amino acid, inosine, coenzyme A, cytidine triphosphate, vitamin C. Common drugs combined with Xiyanping can be divided into 5 categories: nutrition support therapy (vitamin C, compound amino acid) , coenzymes (coenzyme A, cytidine triphosphate, inosine), expectorants and antiasthmatics (ambroxol, salbutamol, doxofylline), hormones (dexamethasone, budesonide), antibiotics (mainly cefminox). The main combined medicines mostly conformed to the regularity for drugs treating lung infection. In addition, there were two most common medical combination models: the model for Xiyanping combined a single medicine is Xiyanping + nutrition support therapy, while the model for Xiyanping combined two or more than two medicines is Xiyanping + nutrition support therapy + coenzyme. Pharmacologically, Xiyanping is mostly combined with western medicines with similar pharmacological effects to substitute or supplement the antibiotic effect in treating lung infection. However, further studies shall be conducted for the safety and rationality of the combined medication based on clinical practices, in order to provide reference for clinical medication.
Adult ; Anti-Bacterial Agents ; administration & dosage ; Ascorbic Acid ; administration & dosage ; Cephamycins ; administration & dosage ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Hospital Information Systems ; Humans ; Lung Diseases ; drug therapy ; Male ; Middle Aged ; Young Adult

OBJECTIVETo explore a new experimental method for screening of allergens in post-market traditional Chinese medicine injections by confirming allergens in Huachansu injection.

METHODFirst of all, the serum of patients allergic to Huachansu injection were collected, at the same time, the dubious allergen was conjugated to bovine serum albumin (BSA) by EDC coupling procedure to form complete antigen (BNP-BSA), which makes it possible to reproduce the allergic reaction of Huachansu injection in vitro. The histamine liberation ratio, the level of TNF-alpha and Histamine released from RBL-2H3 mast cell were detected; the above data were compared with those obtained in vivo.

RESULTThe difference of the histamine liberation ratio, the levels of TNF-alpha and histamine of the resibufogenin-BSA group, group of patients allergic to Huachansu injection were not significant compared with those of normal control group. However, there were significant difference in those data among the cinobufagin-BSA group, the blank control and normal control group (P<0.05).

CONCLUSIONThe allergen in the serum collected from patients allergic to Huachansu injection is resibufogenin.

Allergens ; adverse effects ; immunology ; Amphibian Venoms ; adverse effects ; immunology ; Animals ; Anura ; Bufanolides ; adverse effects ; immunology ; Drug Hypersensitivity ; immunology ; Histamine Release ; Humans ; Mast Cells ; immunology ; Medicine, Chinese Traditional

To control the quality of Humulus scandens, the quality standard was established in this study. According to the method recorded in the Appendix of Chinese Pharmacopoeia (2010 Edition) , the water and ash inspections were carried out. The component luteoloside and cosmosiin in Humulus scandens were identified and assayed by TLC and HPLC. The results showed a strong characteristics microscopic of Humulus scandens, and trichoromethane-methanol-formic acid (10: 3: 0. 3) as the mobile phase of TLC, the spots at 365 nm with a UV lamp was clear. The 16 batches of samples were analyzed by HPLC with a gradient elution of acetonitrile and phosphate solution (0.2%) at a flow rate of 1.0 mL · min(-1) and detected at 350 nm. The content of luteoloside was 0.015%- 0.651% (average 0.148%); the content of cosmosiin was 0.003%-0.118% (average 0.036%). The linear calibration curve of luteoloside and cosmosiin was acquired in the ranges of 0.011-0.364 g · L(-1) (r = 1.000 0) and 0.003-0.096 g · L(-1) (r = 1.000 0), respectively. The average recovery was 100.5% and 98.5%, respectively. The methods are convenient and reliable, which can be ap- plied for quality assessment of Humulus scandens.
China ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; analysis ; standards ; Humulus ; anatomy & histology ; chemistry ; Quality Control

Objective To investigate the effects of puerarin on the expression of human umbilical vein endothelial cells (HUVECs) tissue factor (TF) and tissue factor pathway inhibitor (TFPI) induced by oxidized low-density lipoprotein (ox-LDL).Methods After HUVECs were incubated with different concentrations of puerarin and 50 mg/L ox-LDL,the expression of TF and TFPI mRNA and protein were detected by real-time fluorescent quantitative PCR and Western blot respectively.Results Compared with control,treatment with ox-LDL caused the augment of TF mRNA and protein expression (P<0.01),and the decrease of TFPI mRNA and protein expression.However,50,100,and 200 μmol/L puerarin blunted the augment of TF mRNA and protein expression and weakened the inhibition of TFPI mRNA and protein expression induced by ox-LDL(P<0.01).Conclusions Puerarin reduces HUVECs TF and TFPI mRNA and protein induced by ox-LDL.

AIM: To explore the role of phosphatidylinositiol 3-kinase/protein kinase B/endothelial nitric oxide synthase (PI3K/Akt/eNOS) signaling pathways in the inhibitory effects of puerarin on oxidized low-density lipoprotein (ox-LDL)-induced tissue factor (TF) expression in vascular endothelial cells.METHODS: The mRNA expression of TF was detected by real-time fluorescent quantitative PCR.The protein levels of TF and Akt was determined by Western blot.The content of the nitric oxide (NO) was measured by nitrate reduction method.RESULTS: Compared with control group, incubating endothelial cells with ox-LDL significantly induced TF expression at mRNA and protein levels and the dephosphorylation of Akt protein, and decreased NO production.Incubation of the endothelial cells with puerarin for 1 h and then treatment of the cells with ox-LDL decreased the TF expression at mRNA and protein levels, increased Akt protein phosphorylation and intracellular NO content.Co-incubation of the endothelial cells with PI3K inhibitor LY294002 and puerarin for 1 h and then treatment of the cells with ox-LDL augmented the TF expression at mRNA and protein levels and the Akt protein dephosphorylation, and decreased NO production.Co-incubation of the endothelial cells with eNOS inhibitor NG-nitro-L-arginine methyl ester (L-NAME) and puerarin significantly decreased the inhibitory effect of puerarin on ox-LDL-induced TF expression at mRNA and protein levels in the endothelial cells, and reduced Akt protein phosphorylation and NO production.CONCLUSION: Puerarin inhibits ox-LDL-induced TF expression at mRNA and protein levels in the human umbilical vein endothelial cells via activation of PI3K/Akt/eNOS signaling pathway.