Objective To investigate the transfection and expression of pcDNA3 1-VEGF 165 plasmid in mammalian cells. Methods pcDNA3 1-VEGF 165 plasmid was transfected into CHO cells by electroporation and the positive clones were screened by G418. The expression of VEGF 165 gene in the transfected CHO cells were detected by RT-PCR, ELISA and western blotting analysis. The biologic activities of VEGF 165 secreted by the transfected CHO cells were measured by Mile's assay and MTT method. Results G418-resistant CHO cell clones were obtained. RT-PCR, ELISA and western blotting analysis showed that there were the transcription and translation of VEGF 165 gene in anti-G418 CHO cells. The mile's assay showed that the expressed VEGF 165 could increase the vessel permeability, and MTT assay proved that it could promote the proliferation of endothelial cells. Conclusion pcDNA3 1-VEGF 165 plasmid could express VEGF 165 with biological activities in mammalian cells.

ObjectiveTo evaluate prognostic factors affecting local tumor progression after radiofrequency ablation (RFA) of hepatocellular carcinoma (HCC).MethodsA total of 246 HCC patients (343 lesions) underwent RFA treatment in our department and were enrolled into this study.The average tumor size was 3.7 cm ( range 0.9 ～ 3.7 cm).Regular follow-up with enhanced CT was performed to evalutate the treatment results.Kaplan-Meier model and log-rank test were used in univariate analysis and COX regression model was used in multivariate analysis to identify risk factors for local tumor progression.ResultsThe local tumor progression rate was 11.4％ (39/343 lesions),and the average time from initial RFA to local tumor progression was 12.0 months.Univariate analysis indicated tumor size ( P ＜0.001 ),close to intrahepatic vessels ( P ＜0.001),tumor boundary ( P =0.020),pathological grade( P =0.010) and CEUS before RFA ( P =0.001) as risk factors for local progression.The following factors were identified as independent prognostic factors for local tumor progression by multivariate model:tumor size (P ＜ 0.001),isolated or close to intrahepatic vessels( P ＜0.001) and CEUS before RFA(P =0.018).ConclusionsTumor size,CEUS before RFA and close to intrahepatic vessels are the most important factors for local progression after RFA.Being awaring of possible risk factors for local tumor progression may increase the treatment efficacy and help to promote the use of RFA technique.

Objective To investigate the effect of sustentaculum tali screw placement on outcomes following open reduction and internal fixation (ORIF) of intraarticular calcaneal fractures.Methods A retrospective analysis was made on 139 patients with intraarticular calcaneal fractures treated by ORIF from April 2008 to January 2012.According to Sanders classification,there were 9 patients with type Ⅱ fractures,87 with type Ⅲ fractures and 43 with type Ⅳ fractures.Fracture reduction followed by placement of calcaneal anatomic plate or anatomic locking plate was performed via calcaneal lateral L-shape incision.All the patients received X-rays and CT scans within postoperative 2 weeks.Placement of screws in sustentaculum tail was detected by CT volume rendering to visualize implants combined with multiplanar reconstruction imaging (MPR).Patients were divided into accurate fixation group (Group A,n =28),marginal fixation group (Group B,n =58),and non-fixation group (Group C,n =53) based on radiological results.Intra-and peri-operative parameters,time to partial weight-bearing,time to full weight-bearing,fracture healing time,and clinical outcome were compared among groups.Functional outcome was assessed using American Orthopedic Foot and Ankle Society (AOFAS) Ankle Hindfoot Scale.Results Postoperative X-rays indicated accurate fixation of sustentaculum tail in 64 patients,marginal fixation in 22,and non-fixation in 53,but CT images manifested accurate fixation in 28 patients,marginal fixation in 58,and non-fixation in 53.Group A had operation duration of (93 ±11) min and intraoperative blood loss of (252 ± 27) ml,longer or higher than those in Group B [(85 ±8) min,(194±16) ml] and Group C [(57±6) min,(136 ±13) ml] (P＜O.05).There was no significant difference in mean hospital stay among the three groups (P ＞ 0.05).Ninety-two patients were followed-up for 12-38 months (mean 18.5 months) and 85 patients completed foot and ankle exercise as planned.Moreover,no significant differences were observed among groups concerning time to partial and full weight-bearing,fracture healing time and AOFAS score at the final follow-up (P ＞ 0.05).Conclusion Sustentaculum tali screw placement has no apparent effect on the short-term outcome of surgical treatment for calcaneal fractures.

Objective To clone and express bradyzoite antigen 1(BAG1) gene of T. gondii,and analyze the immunoreactivity of the recombinant product. Methods The differentiation of T. gondii RH strain tachyzoites into bradyzoites was induced in vitro,and the coding sequence of BAG1 was amplified from bradyzoites by RT-PCR. The PCR product was analyzed by sequencing. The BAG1 coding sequence was further subcloned into the plasmid pET32a(+). The plasmid pET32a(+)-BAG1 was then transformed into BL21(DE3) to express after IPTG induction. The expression product was purified with Ni-NTA agarose and the purified BAG1 was further analyzed by Western blotting and ELISA. Results BAG1 cDNA was amplified from bradyzoites. After IPTG induction,BAG1 was expressed in a fusional form in E. coli. Western blotting showed that the purified recombinant protein could be specifically recognized by sera from mice chronically infected by T. gondii B36 strain. ELISA showed that the positive rate of T. gondii IgG antibodies of 350 human sera detected by the recombinant BAG1(17.4%) was higher than by recombinant SAG1 (12.6%)(P

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine ; analogs & derivatives ; pharmacology ; Animals ; Cardiomyopathy, Hypertrophic ; drug therapy ; metabolism ; Connexin 43 ; metabolism ; Disease Models, Animal ; Male ; Myocardium ; metabolism ; Rats ; Rats, Wistar

The purpose of this study is to investigate the transdermal delivery characteristics of Gentiana macrophylla complex components system through different parts of the skin under micro-needles conditions. Two-chamber diffusion cells were used, different parts of isolated skin and micro-needle pretreated isolated mouse skin were applied separately, high performance liquid chromatography (HPLC) similarity evaluation methods were used to evaluate transdermal delivery characteristics of Gentiana macrophylla complex components system on receiving pool and the permeation rate and penetration amount of Gentiopicroside at different parts of mouse skin. In the 24 h, the similarity between receiving fluid which was on passive transdermal delivery and micro-needle transdermal delivery conditions and original fluid were ranged from 83.0% to 98.9%; By the micro-needle pretreatment with different parts of the mouse skin, the time that Gentiana macrophylla complex components system though abdominal skin to the receiving fluid which reached 90% similarity compared with that of original fluid was 4 h, which was 18 h at back skin and 12 h at neck skin separately. Micro-needles can be used as the ideal ingredients for traditional Chinese medicine complex transdermal delivery; transdermal absorption time delay could be greatly reduced and its bioavailability was improved. The permeation rate and similarity to original liquid of Chinese medicine complex components increased significantly in the abdominal skin relative to the neck and back skin under micro-needle conditions.

Objective To evaluate the clinical effect of the intertrochanteric fractures with or without lateral femoral wall fractures using proximal femoral nail antirotation (PFNA).Methods From May 2008 to June 2011,102 patients with intertrochanteric fractures were treated with PFNA.In accordance with the preoperative three dimensional CT reconstruction(3D CT) images,the group A included 41 cases with lateral femoral wall fractures,and the other 61 cases with an intact lateral wall were in group B.According to the AO/OTA classification,there were 5 cases in 31-A2,36 in 31-A3 in group A,and 61 in 31-A2 in group B.The operative time,operative blood loss,average length of stay,postoperative X-ray images,and 3D CT images were collected for each patient.Time of partial weight-bearing,full weight-bearing and fracture healing were also recorded.Clinical evaluation was made using the functional recovery scale (FRS) of hip fractures.Results The mean operation time was 56±8 min in group A vs 45±6 min in group B; the mean blood loss was 238±21 ml vs 175±11 ml; the average length of stay was 17±3 days vs 15±3 days.On the postoperative radiography,the blowout of lateral trochanteric wall only occurred in 8 (19.5％,8/41) cases in group A and 3 (4.9％,3/61) in group B.According to the postoperative 3D CT,the similar findings were seen in 36 (87.8％.36/41) cases in group A and 45 (73.8％,45/61) in group B.Eighty-two cases were followed up for 6 to 35 months (mean.19.5 months).The mean FRS score was 64.2±4.8 points in group A and 76.5±7.9 points in group B.Conclusion When treating unstable intertrochanteric fractures,iatrogenic fractures in lateral trochanteric wall could be easily caused with using PFNA.3D CT could effectively evaluate iatrogenic trauma in the intertrochanteric fractures.

Objective To discuss the safety and effectiveness of the drainage of perinephric abscess with retroperitoneal laparoscopy.Methods The clinical data of 12 cases undergoing the drainage of perinephric abscess with retroperitoneal laparoscopy were analyzed retrospectively.There were 3 males and 9 females,with a mean age of 47 years.All the 12 cases presented with lumbar pain and fever,7 cases had chill,5 cases had irritative symptom of bladder,and 4 cases had nausea and vomiting.The average course was 9 d,and the average diameter was 6.4 cm.Four cases were in the left side,and 8 in the right side.The main complications included type-2 diabetes (5 cases),type-1 diabetes (1 case),kidney stones (2 cases) and systemic lupus erythematosus (1 case).All the 12 cases were treated with antibiotics.The indexes were recorded,including operative time,intraoperative blood loss,purulent fluid volume,postoperative indwelling time of the drainage tube,postoperative time of body temperature returned to normal,postoperative hospital stay,and complication rate.Results All the 12 operations were successful without peritoneal damage,kidney damage or other complications.The mean operative time was 56 (48-95) min,the mean intraoperative blood loss was 127 (60-150) ml,the mean intraoperative purulent fluid volume drawed out was 128 (120-230) ml,the mean postoperative indwelling time of the drainage tube was 9 (6-12) d,the mean postoperative time of body temperature returned to normal was 3 (2-4) d,and the mean postoperative hospital stay was 10 (7-12) d.All the patients were followed up for an average of 6.5 (6-12) months,no recurrence occured.Conclusions The drainage of perinephric abscess with retroperitoneal laparoscopy has the advantages of definite effectiveness,complete drainage,little injury and rapid recovery.The drainage of perinephric abscess with retroperitoneal laparoscopy by skilled urologists is safe.

Objective To examine the relationship between gender and intraventricular hemorrhage (IVH) in very low birth weight infants (VLBWI) and extremely low birth weight infants (ELBWI).Methods From January 1,1999 to December 31,2012,data on VLBWI and ELBWI,who were admitted to the neonatal intensive care unit of Yuying Children's Hospital within 14 d after birth,were retrospectively collected.The Chi-square test and t test were used to compare neonatal outcomes between male and female infants.The Logistic model was used to analyze the risk factors for IVH.Results A total of 1 008 cases were enrolled,including 615 males and 393 females,895 VLBWI and 113 ELBWI.The incidence of IVH was 15.1％ (152/1 008) and the incidence of severe IVH was 8.4％ (85/1 008).Compared with females,males had a higher total incidence of IVH [17.2％ (106/615) vs 11.7％ (46/393),x2=5.728,P＜0.05] and severe IVH [9.8％ (60/615) vs 6.4％ (25/393),x2=3.896,P＜0.05].These differences were also seen in VLBWI with a birth weight of 1 250 to 1 499 g [IVH:13.7％ (47/344) vs 7.8％ (17/217),x2=4.473,P=0.034; severe IVH:7.6％ (26/344) vs 2.8％ (6/217),x2=5.684,P=0.017].Logistic regression analysis showed that the risk factors for IVH were as follows:gestational age ＜28 weeks (aOR=2.012,95％CI:1.288-3.143,P＜0.05),neonatal respiratory distress syndrome (aOR=l.584,95％CI:1.007-2.492,P＜0.05),invasive mechanical ventilation (aOR=2.743,95％CI:1.826-4.121,P＜0.05),electrolyte disturbance (aOR=2.128,95％CI:1.092-4.149,P＜0.05) and periventricular leukomalacia (aOR=2.901,95％CI:1.312-6.416,P＜0.05),but not male sex (aOR=1.351,95％CI:0.917-1.991,P=0.128).The risk factors for severe IVH were gestational age ＜28 weeks (aOR=2.200,95％CI:1.305-3.708,P＜0.05),invasive mechanical ventilation (aOR=4.714,95％CI:2.809-7.911,P＜0.05) and electrolyte disturbance (aOR=2.232,95％CI:1.047 4.759,P＜0.05),but not male sex (aOR=1.361,95％CI:0.823 2.252,P=0.247).Conclusions Male VLBWI and ELBWI have a higher incidence of IVH and severe IVH,but male sex is not a risk factor for IVH or severe IVH.

BACKGROUND:Animal studies have shown that cauda equina compression can induce apoptosis of lumbosacral spinal cord anterior horn motor neurons.
OBJECTIVE:To explore the pathological change in lumbosacral spinal cord after acute cauda equina compression in dogs.
METHODS:A total of 27 dogs were randomly divided into nine groups, with three dogs in each group. There were one normal control group, seven experimental groups and one sham surgery group. In the experimental group, an empty water sac was implanted above epidural fat below L6 vertebral plate. Compression was given by injecting water at 4, 8, 12, 24, 48, 72 and 168 hours. In the sham surgery group, an empty water sac was implanted, but compression was not given. At the time of compression, the spinal cord sent out by cauda equina nerve and adjacent to the head end was subjected to histopathological examination.
RESULTS AND CONCLUSION:(1) Results of light microscope:at 4-48 hours of compression, spinal cord anterior horn motor neurons did not alter. At 72 hours, motor neurons became smal , cel membrane shrank and separated from surrounding tissues. Cel s were homogenous and darkly stained. At 168 hours, motor neurons disappeared, but spinal cord sections of the adjacent head end did not shown abnormal motor neurons in the spinal cord anterior horn. (2) Results of electron microscope:at 12 hours, spinal cord tissue began to swel , and the swel ing aggravated with prolonged time of compression. The swel ing of glial cel s was apparent. At 168 hours, myelin sheath structure dissolved;axons showed vacuolization;axoplasm spil ed, and exhibited inflammatory injury-like changes. (3) Apoptotic results of spinal cord anterior horn motor neurons:apoptosis appeared at 12 hours of compression, became increased, and showed an increased trend at 168 hours.