BACKGROUND: Double gene transfection using bone morphogenetic protein 2 (BMP2) and vascular endothelial growth factor 165 (VEGF165) for bone marrow stromal stem cells (BMSCs) to induce osteogenesis provides experimental basis for the study on tissue engineering bone. OBJECTIVE: To investigate the ability of BMP2 and VEGF165 double gene modified rat BMSCs to induce osteogenesis. METHODS: BMSCs were isolated from the femur and tibia of four 4-week-old Sprague-Dawley rats by whole bone marrow adherent culture. Passage 3 BMSCs were randomized into five groups: non-transfection group, empty plasmid group, BMP2 transfection group, VEGF165 transfection group, BMP2 and BMP2/VEGF165 transfection group (co-transfection group). Then, western blot assay was used to detect expression of BMP2 and VEGF165 at 48 hours after transfection, and the activity of alkaline phosphatase was detected in each group at 7 days after transfection. RESULTS AND CONCLUSION: Highly expressed BMP2 in BMP2 and co-transfection groups and highly expressed VEGF165 in VEGF165 and co-transfection groups were found after transfection. The expression of BMP2 or VEGF165 in the co-transfection group was significantly higher than that in the BMP2 or VEGF165 transfection group after transfection, respectively (P < 0.05). Western blot results showed that the activity of alkaline phosphatase was ranked as: co-transfection group > BMP2 transfection group > VEGF165 transfection group > empty plasmid group and non-transfection group. There was a significant difference in the activity of alkaline phosphatase between co-transfection group and any of single gene transfection groups (P < 0.05). To conclude, BMP2/VEGF165 co-transfection promotes the ectopic osteogenesis of BMSCs.

Objective To explore the efficacy of proximal femur locking plate(PFLP) for the treatment of unstable femoral intertrochanteric fractures in elderly patients.Methods Data of 120 cases of patients with unstable femoral intertrochanteric fractures who were treated by PFLP were retrospectively analyzed.There were 56 males and 64 were females,aging from 62 to 78 year-old (average,69.4 year-old).85 cases were left hip,and 35 cases right hip.According to AO fracture classification,there were 12 cases of A2.2,23 A2.3,34 A3.1,29 A3.2,and 22 A3.3.According to Evans intertrochanteric fractures classification:there were 93 cases of type Ⅲ,19 type Ⅳ,and 8 type Ⅴ.Operation time,length of hospital stay,fracture healing time,postoperative complications and hip joint Harris scores were recorded.Results All patients were followed up for 8-14 months (average,12.4 months).Their operating time were 51-80 min (average,63.1 min);X-ray exposure were 8-15 times (average,11.6 times);intraoperative blood loss were 70-120 ml (average,92.7 ml);postoperative drainage volume were 50-100 ml (average,61.7 ml),and length of hospital stay were 7-14 d (average,10.2 days).The healing time ranged from 12 to 32 weeks (20.4 weeks average).14 cases had delayed union,and healing time was 8-10 months.In addition,4 cases had deep vein thrombosis (after thrombolysis therapy the vein was recanalized);2 had internal fixation broken,who were then treated with PFNA Ⅱ,and bone healed after 6 months;5 had hip varus,who were then treated via hip replacement,and no hip prosthesis fracture were found.No other complications occurred in the remaining patients.At the latest follow-up,according to Harris hip score:there were 38 cases with excellent results,71 good,9 fair,and 2 poor,and the rate of excellent and good was 90.8％ (109/120).Conclusion PFLP for the treatment of unstable femoral intertrochanteric fractures has advantages of less invasion,less complication,reliable fixation and high fracture healing rate,thus it is suitable for unstable intertrochanteric fractures in elderly patients.

Objective To investigate the analgesic effect of oxysophoridine(OSR)and the influence of verapamil(Ver)on the antinociception of OSR when two drugs were co-administrated in mice.Methods The number of writhing within 15 min after ip different doses of OSR was observed in painful mouse mo- dels caused by acetic acid.The hot plate method was used to assess nociceptive sensitivity of CaCl_2 and Ver before ip OSR.Nitric oxide(NO)in serum was measured by spectrophotometry.Results The number of writhing was decreased and the latency of licking the hind paws was prolonged in a dose-dependent manner after ip OSR.The antinociception of OSR could be antagonized by CaCl_2 and enhanced by Ver.No inter- ference was detected in serum volume of NO.Conclusion These results suggest that OSR can antagonize the acute pain caused by acetic acid and hot plate in a dose-dependent manner in mice.Calcium channel blocker could enhance the effect of OSR.

Background Efficient and lowcost way to isolate keratocytes is helpful for research on cornea.Either relatively expensive or inefficient is the shortage of those means now applied,while raising the keratocytes through passage will change the phenotype of them quickly.Our aim is to approach the way getting keratocytes effectively utilizing modified two step enzymatic digestion by type I collagenase. Objective To evaluate the effect of isolating the bovine keratocytes utilizing two step enzymatic digestion and observe the morphological changes of the keratocytes during cultivation in vitro. Methods Keratocytes were isolated from bovine corneas using 0.5 mg/mL and 1 mg/mL type I collagenase digestion.The harvesting rate and viability rate of the primary keratocytes were evaluated.During the primary cultivation in vitro,the morphological changes of the keratocytes and their F-action distribution were observed.Results(1)The extracellular matrix of the bovine corneas were almost dissolved by the two step enzymatic digestion,followed the keratocytes completely isolated from the solid matrix.The amount of the harvested keratocytes from each cornea was(2.11±0.15)X106 on average while the viability rate was(91.69±3.73)% and the inoculation rate Was(81.20±1.25)%.(2)The primary keratocytes attached and spreaded out with dendritic and stellate morphology.After 3 days cultured,the branches of the keratocytes were contacting and formed networks.The F-actin detected by phalloidin binding showed a limited cortical localization. Conclusion (1)The method of two step enzymatic digestion can make the extracellular matrix of bovine cornea stroma completely degraded with the advantages in high efficiency of harvesting keratocytes and high cell viability and relatively simple manipulation. (2) The primary bovine keratocytes have dendritic morphology and with limited F-action distribute in cellular cortex.

Background The incidence of posterior capsular opacification (PCO) is increasing with the growing of cataract surgery rate.Recent researches provend that disintegrin has inhibitory effect on PCO,and echistatin is one of the disintegrin prime families.Objective This study was to investigate the effects of disintegrin and echistatin on proliferation,adhestion and migration in human lens epithelial cells (LECs) line (SRA01/04).Methods Human LECs line at logarithmic growth phase was used in the study.Cells were cocultured with medium and different concentrations of echistatin (0,2.5,5.0,7.5,10.0,15.0,20.0 mg/L) for different time.The proliferative inhibitory rates of LECs were detected by MTT method 24,48 and 72 hours after cultured.Anti-adhesion effect of echistatin were analyzed by the same assay in 90 minutes.Cell scratching test was performed to evaluate the migration ability of LECs.The width of the scratch was recorded in the culture plate covered with cells under an inverted microscope.After being cultured for 24 hours and 48 hours with echistatin,cell migration distances was examined.Results Compared with the 0 mg/L echistatin group,cells proliferation was obviously inhibited.After cultured with 2.5,5.0,7.5,10.0,15.0,20.0 mg/L echistatin,the proliferation inhibitory rate was 2.6％,15.4％,21.2％,34.7％,46.1％,58.2％ at 24 hours;6.6％,21.9％,38.2％,50.0％,60.7％,76.9％ at 48 hours and 9.8％,29.0％,46.6％,63.4％,69.1％,92.4％ at 72 hours,respectively.The absorbance value (A) in the 5.0,7.5,10.0,15.0,20.0 mg/L groups were significantly lower than that in the 0 mg/L group (P＜ 0.05).With the prolongation of acting time of Ecs,the A value of the cells was gradually reduced,with statistically significant difference (P＜0.05).The adhesion inhibitory rate was 2.6％,15.0％,26.1％,35.3％,45.2％ and 54.5％ in the 2.5,5.0,7.5,10.0,15.0,20.0 mg/L group,respectively.Compared with the result in the 0 mg/L group,the A value in the 5.0,7.5,10.0,15.0,20.0 mg/L group was statistically significant (P＜0.05).After cultured for 24 hours and 48 hours,cell migration distance shortened in the 5.0,7.5,10.0,15.0,20.0 mg/L group,showing a statistically significant difference among them (P＜0.05).Cell migration distance was gradually shortened with the lapse of action time of Ecs with the significant difference (P ＜ 0.05).Conclusions echistatin has inhibitory effects on proliferation,adhestion and migration for human LECs in vitro in time-and dose-dependent manner.It is inferred that echistatin may play a role in the prevention and treatment of PCO.

This work was mainly studied the effects of the four alkaloids from Coptidis Rhizoma on the mouse peritoneal macrophages in vitro and preliminarily discussed the regulating mechanisms. The effect of alkaloids from Coptidis Rhizoma on the vitality of macrophages was measured by the MTT assay. The effect of alkaloids on the phagocytosis of macrophages was determined by neutral red trial and respiratory burst activity was tested by NBT. The expressions of respiratory-burst-associated genes influenced by alkaloids were detected by qRT-PCR. The conformation change of membrane protein in macrophages by the impact of alkaloids was studied by fluorospectro-photometer. Results showed that the four alkaloids from Coptidis Rhizoma could increase the phagocytosis of macrophages in different level and berberine had the best effect. Berberine, coptisine and palmatine had up-regulation effects on respiratory burst activity of mouse peritoneal macrophages stimulated by PMA and regulatory activity on the mRNA expression of PKC, p40phox or p47phox, whereas the epiberberine had no significant influence on respiratory burst. Moreover, alkaloids from Coptidis Rhizoma could change the conformation of membrane protein and the berberine showed the strongest activity. The results suggested that the four alkaloids from Coptidis Rhizoma might activate macrophages through changing the conformation of membrane protein of macrophages and then enhanced the phagocytosis and respiratory burst activity of macrophages. Furthermore, the regulatory mechanism of alkaloids on the respiratory burst activity of macrophages may be also related to the expression level of PKC, p40phox and p47phox.
Alkaloids ; pharmacology ; Animals ; Cells, Cultured ; Coptis ; chemistry ; Drugs, Chinese Herbal ; pharmacology ; Female ; Gene Expression ; drug effects ; Macrophages, Peritoneal ; drug effects ; Mice ; Phosphoproteins ; genetics ; metabolism ; Protein Kinase C ; genetics ; metabolism ; Rhizome ; chemistry

Objective To analyze the related factors of femoral head necrosis after internal fixation operation of femoral neck fracture.Methods Clinical data of 524 patients with femoral neck fracture who underwent internal fixation surgery between January 2012 and January 2015 were retrospectively analyzed.Taking the femoral head necrosis as an observable indicator,the clinical data of postoperative femoral head necrosis and those other patients who were treated at the same period were compared.Whether the clinical factors (gender,age,surgical reduction type,fracture type,quality of reduction,post-injury operation time,loading time after operation,and whether to withdraw internal fixation) were in correlation with postoperative femoral head necrosis was analyzed by univariate analysis.Then the statistically significant indicators were integrated into logistic regression analysis to determine the related factors of femoral head necrosis.Results According to inclusion and exclusion criteria,the study group consisted of 212 cases,including 94 males and 118 females;the follow-up time was 2-5 years,and the average follow-up time was 3.6±1.6 years.There were 46 cases with femoral head necrosis and 166 cases without femoral head necrosis after operation.The rate of femoral head necrosis was 21.7％ (46/212).The femoral head necrosis group included 46 cases with 21 males and 25 females;20 cases below 60 years old and 26 cases above 60 years old;17 cases using open reduction and 29 cases using closed reduction;6 cases belong to type Ⅰ/Ⅱ and 40 cases belong to type Ⅲ/Ⅳ according to Garden classification;16 cases got satisfactory reduction while 30 cases with unsatisfactory reduction;30 cases below 48 h and 16 cases above 48 h on post-injury operation time;17 cases below 3 months and 29 cases above 3 months on post-operation loading time;22 cases with internal fixation removal and 24 cases without internal fixation removal.The femoral head without necrosis group included 166 cases with 73 males and 93 females;84 cases below 60 years old and 82 cases above 60 years old;69 cases using open reduction and 98 cases using closed reduction;120 cases belong to type Ⅰ/Ⅱ and 46 cases belong to type Ⅲ/Ⅳ according to Garden classification;160 cases got satisfactory reduction while 6 cases with unsatisfactory reduction;119 cases below 48 h and 47 cases above 48 h on post-injury operation time;70 cases below 3 months and 96 cases above 3 months on post-operation loading time;74 cases with internal fixation removal and 92 cases without internal fixation removal.Univariate x2 analysis suggested that fracture type and quality of reduction were associated with postoperative femoral head necrosis.Other clinical factors (gender,age,surgical reduction type,post-injury operation time,loading time after operation and whether to remove internal fixation or not) were not associated with postoperative femoral head necrosis.The multivariate Logistic regression analysis showed fracture type and quality of reduction were significantly related to postoperative femoral head necrosis.Conclusion Femoral head necrosis after internal fixation operation of femoral neck fracture is affected by many factors.The fracture type and quality of reduction are important factors affecting femoral head necrosis after internal fixation operation of femoral neck fracture.

With the development of treatment, the survival rate of premature infants has significantly increased, especially extremely premature infants and very low birth weight infants. This has led to an increase in incidence of bronchopulmonary dysplasia (BPD) year by year. BPD has been one of the most common respiratory system diseases in premature infants, especially the small premature infants. Arrested alveolar development is an important cause of BPD. Therefore, the mechanism of arrested alveolar development and the intervention measures for promoting alveolar development are the focuses of research on BPD. Selecting the appropriate animal model of BPD is the key to obtaining meaningful results in the basic research on BPD. Based on above, several common methods for establishing an animal model of BPD and the corresponding changes in pathophysiology are summarized and evaluated in order to provide a reference for selecting the appropriate animal model in studies on the pathogenesis, pathophysiology, and prevention and control strategies of BPD.
Animals ; Bronchopulmonary Dysplasia ; etiology ; Disease Models, Animal ; Humans ; Hyperoxia ; complications ; Respiration, Artificial ; adverse effects

A strain was isolated from internal organ of died porcine about 8 weeks with purulent pneumonia,arthritis,pyogenic arthritis and endocarditis in April 2007.Objectives of the study are to confirm the genus of the strain,pathopoiesis,and drug sensitivity.The mainly study methods:the first,the strain was identified by the phenotype and the characteristics of the biochemistry,sequence 16S rDNA genes of the strain was analyzed by molecular biology technology,finally animal experiment and drug sensitivity testing were done.The results of the phenotype and the characteristics of the biochemistry showed that it is greatly similar to Actinomyces hyovaginalis,16S rRNA sequence analysis exhibited the homology achieved to 99.2% com-pared with group III strains of Actinomyces hyovaginalis,and the phyletic evolution analysis also indicated that it has mostly relationship with group III strains of Actinomyces hyovaginalis.Animal experiment dis-covered it has highly pathogenicity to Mus musculus albus;Drug sensitivity testing showed that it is hyper-sensitive to Erycin,Gentamicin and Amikacin.So,the result of the study confirmed that the strain is Actin-omyces hyovaginalis III with the pathogenicity.