BACKGROUND: It is a new tendency to treat central nervous system injury or tumor therapy using the combination of seed cells and gene therapy.OBJECTIVE: To observe the dose-relationship between transfection and expression of rabbit bone marrow mesenchymal stem cell (BMSCs) with adenovirus vector carrying green fluorescent protein (Ad-GFP), and to study its effects on cell biological properties, in addition, to explore the feasibility of using Ad-GFP vector to construct gene modified BMSCs.DESIGN, TIME AND SETTING: A randomized grouping, contrast observation. The experiment was performed at the Southern Medical University between August 2008 and March 2009.MATERIALS: New Zealand white rabbits, irrespective of genders, weighing 2.0-3.0 kg, were selected.METHODS: BMSCs were separated and cultured in vitro, and then the cell immunophenotypes were detected by flow cytometry.The adenovirus was obtained by packaging 293 cells and was used to transfect BMSCs with various liters (1 ×-10~3-1×10~(10) PFU/mL).Cytometry was used to analyze the transfection efficiency.MAIN OUTCOME MEASURES: Cell morphological changes were detected under an invert microscope. The cell proliferation was detected by CCK8 kits. BMSCs transfected with Ad-GFP were induced differentiating into neuron-like cells by adding of β-mercaptoethanol.RESULTS: The surface markers of 3-6-generation BMSCs were negative to CD34 and CD45, but positive for CD29 and CD44.When the virus titers were 1 ×10~7 PFU/mL, the transfection rate was 55%, which were 85% when the virus titers were 1 ×10~9 and1×10~(10) PFU/mL. However, cell pathology phenomenon occurred when the virus titer was 1 ×10~(10) PFU/mL. The fluorescence was strongest expressed at day 7, and it still can be seen at day 28. The BMSCs trasfected with Ad-GFP could differentiate into neuron-like cells under induction of p-mercaptoethanol, with positive neuron-specific enolase.CONCLUSION: Ad-GFP with suitable titers can infect BMSCs effectively with little influence on the biology property or differentiation function. BMSCs can serve as seeds cell in gene therapy field when utilizing ad-GFP vector system.

Objective To investigate the curative effect of using new irrigation pump in acute hematogenous osteomyelitis.Methods Forty patients with acute hematogenous osteomyelitis were divided into the control group and observation group,20 cases in each group.The control group adopted the conventional lavage for drainage,while the observation group used self-made irrigation pump.Then the clinical curative effect and cure time were compared between the two groups.Results The total effective rate in the observation group was 100.00 ％,which was significantly higher than 80.00 ％ in the control group(P＜0.05),and the difference was statistically significant(P＜0.05).The cure time in the observation group was(10.13 ± 8.03)d,which was significantly shorter than (18.14-±5.43)in the control group,and the difference was statistically significant(P＜0.05).Conclusion The curative effect of new type irrigation pump in acute hematogenous osteomyelitis is more acurate than that of conventional lavage for vacuum darianage.

BACKGROUND:Many scholars have developed a variety of dynamic elastic spine fixator. After biomechanical research, animal experiments and clinical application found that no one elastic spine fixator was general y recognized clinical y. OBJECTIVE:To compare the stress difference between“X”-shaped spine dynamic fixation and traditional pedicle screw fixation. METHODS:Three-dimensional finite element models of“X”-shaped spine dynamic fixation and traditional pedicle screw fixation were established according to adult spine imaging data. Mechanical differences in vertical compression, flexion, extension, lateral bending and rotation were compared between the two groups. RESULTS AND CONCLUSION:The stress at vertical compression was lower than that at flexion, extension, lateral bending and rotation in both groups. The stress at“X”-shaped spine dynamic fixation mainly focused on“X”-shaped connecting rod, but the stress of traditional pedicle screw fixation mainly focused on conjunction of screw-rod. Moreover, the stress of the screw of“X”-shaped spine dynamic fixation was significantly less than that of traditional pedicle screw fixation (P<0.001). These results suggest that“X”-shaped spine dynamic fixation system can share stress of screws and reduce the postoperative stress concentration compared with traditional pedicle screw fixation.

Objective To optimize the extraction technology of Taxus x media by using the contents of Paclitaxel and 10-deacetylbaccatin(10-DAB),two representative active diterpene alkaloids of taxane type from T.x media,as evaluation standard.Methods The smashing tissue extraction(STE)of Paclitaxel and 10-DAB from T.x media,was investigated by comparing with ultrasonic extraction(UE)which was one of the modern technologies of extraction.Results STE was more efficient than UE,and the contents of 10-DAI3 and Paclitaxel in the extracts obtained by STE were higher than those by UE.Conclusion STE is a fast,high-performance,and energy-saving technology for the extraction of diterpene alkaloids of taxane type.STE also provides a simple,component-safe,workable,and highly efficient method for the extraction of active natural product.

ObjectiveTo investigate the clinical effects of posterior atlantoaxial fusion fixation in treatment of old atlantoaxial injury secondary to atlantoaxial dislocation.MethodsA retrospective analysis was carried out on 16 patients ( 14 males and 2 females) with old atlantoaxial injuries secondary to atlantoaxial dislocations managed with posterior atlantoaxial fusion fixation from March 2008 to March 2012.The time from injury to operation lasted for 3-36 months ( average 10.5 months).Posterior atlantoaxial transpedicular fixation was performed in 13 patients including 10 patients with old odontoid fractures and three with old traumatic transverse ligament disruptions of the atlas combined with atlantoaxial dislocations.Also,posterior atlantal arch transpediclar fixation combined with axial pedicle screw fixation was performed in three patients who had old odontoid fractures combined with atlantoaxial dislocations.All patients had simultaneous autogenous bone grafting between atlas and axis during reduction and fixation.The preoperative and postoperative Japanese Orthopaedic Association (JOA) scores were compared.Follow-up X-ray films and CT was performed to evaluate the atlantoaxial reduction and fusion.ResultsAll the patients were followed up for 9-18 months ( mean 13 months).None of the patients had spinal cord or vertebral artery injuries.Follow-up CT showed that two patients had partial penetration of one side axial pedicle screws into transverse foramen without nerve and blood vessel injuries.Clinical symptoms obtained different degree of improvement.The postoperative JOA scores ranged from 13 to 16 points ( mean 14.8 points) and the improvement rate of JOA was 71％-92％ ( mean 82％ ).The X-ray films and CT showed sound bone fusion,with good location of screws but with no signs of atlantoaxial instability or loss of reduction,or loosening or breakage of the screws.ConclusionPosterior atlantoaxial fusion fixation can effectively reconstruct atlantoaxial stability,improves neurologic function of spinal cord and has reliable curative effects.

Objective To compare the effects of two surgical procedures via comparison on tibial fractures with/without proximal fibula fractures ,and to explore the reference importance of proximal fibula fractures in surgery for complex tibial plateau fracturesinvolvingposterior and lateral spine .Methods Totally 69 cases of patients with complex tibial plateau fracturesinvolving‐posterior and lateral spine (Schatzker Ⅴ and Ⅵ) were selected ,who accepted treatment between January 2008 and October 2012 . The patients were screed according to the quality of fracture reduction immediately after operation .39 cases were screened whose scores were excellent and the general condition was similar .The patients were divided into group A1 ,A2 and B1 and B2 according to the whether there was proximal fibula fractures and whether accepted posterior lateral support steel plate:group A1 (n=10):without proximal fibula fractures but with posterior lateral support steel plate;group A2 (n=11):without proximal fibula fractures and without posterior lateral support steel plate;group B1 (n=9):with proximal fibula fractures and with posterior lateral support steel plate;group B2 (n=9):with proximal fibula fractures but without posterior lateral support steel plate .According to the knee scoring system of American Hospital for Special Surgery (HSS) ,a total of 39 cases of patients were followed up for 12-14 months (average 12 .8 months) to evaluate the recovery condition of knee function .And correlations between the scores of the 4 groups were compared .Results There were statistically significant differences between group B1 and group B2 on HSS scores (P<0 .01) ,but there were not between group A1 and A2 (P>0 .05) .Conclusion For patients with complex tibial plateau fracturesin‐volvingposterior and lateral spine and with proximal fibula fractures ,posterior lateral support steel plate can achieve more satisfac‐tory rehabilitation function .

Objective To explore the relationship between plasma homocysteine levels and diabetic retinopathy (DR).MethodIn 40 DR patients and 63 non-DR patients,the indexes including plasma homocysteine concentrations,duration of diabetes,ages,blood pressure,preprandial blood glucose(PBG),plasma GHbA,e,plasma lipids,serum folie acid and semm vitamin B12 were analyzed.ResultsPlasma homocysteine level was significantly higher in DR patients (12.9μmol/L) than that in non-DR patients (7.8μ.mol/L) (P＜0.01),and the relationship remained significant correlation after adjusted for duration of diabetes,GHbA1c,ages,serum folic acid and serum vitamin B12(OR = 1.23,P＜0.05).ConclusionHyperhomocysteinimia is a risk factor and an independent predieator of DR.

Objective To investigate the the operative time of two stage soft ureteroscope lithotripsy for hemorrhagic embolism after percutaneous nephrolithotomy(PCNL).Methods The clinical data of 1 patient with massive hemorrhage after PCNL who treated with superselective renal artery embolization and ureteral soft lens were analyzed.Results The patient with postoperative bleeding after PCNL was treated with superselective renal artery branch embolization,after 30d embolization,the holmium laser lithotripsy under the soft ureteroscope for ureteral calculi was performed,and the renal pelvis mucosa smooth,no bleeding and scar formation were intraoperative visible.After operation,the stone was removed basically,and the double J tube was removed 2 weeks after operation.The patient had no special discomfort and the renal function was normal.Conclusion Postoperative 14-30d is a relatively safe time to perform flexible ureteroscopic lithotripsy for super selective renal artery embolization in the treatment of patient with massive hemorrhage after PCNL.

Objective To investigate the associations between the single nucleotide polymorphisms of TERT rs2736098, CLK3 rs11543198 and bladder cancer. Methods 201 bladder cancer cases and 200 healthy controls were included in the research, and the genotypes of TERT rs2736098 and CLK3 rs11543198 were determined using the PCR-RFLP method. Relationship between genotypes and bladder cancer risks was investigated. Results There were statistical significance in the rs2736098 genotype frequencies and allele frequencies between cases and controls (χ2= 6.973, P = 0.031; χ2= 7.412, P = 0.006). Compared with the individuals with the GG genotype , the risk of bladder cancer increased 2.069 times with the AA genotype (OR = 2.069, 95%CI: 1.181-3.624, P = 0.011). And there were no significant associations between the rs11543198 genotype frequencies and allele frequencies between cases and controls (χ2 = 0.202, P = 0.904; χ2 = 0.188, P = 0.665). Rs2736098 and rs11543198 genotype distribution in bladder cancer pathologic grade and stage had no statistical significance (P > 0.05). Conclusion Rs2736098 polymorphism is associated with risk of bladder cancer and rs11543198 polymorphism was not associated with risk of bladder cancer.

Objective To explore the relationship between plasma homocysteine levels and diabetic peripheral neuropathy (DPNP). Methods A crossectional analysis was conducted on 227 patients with type 2 diabetes. Peripheral neuropathy was confirmed using electromyography (EMG). The risk factors possibly associated with diabetic neuropathy or plasma homocysteine levels were analyzed in relation to likelihood of occurrence of DPNP. Results Eighty patients with neuropathy and 147 patients without neuropathy were included. Plasma homocysteine levels were significantly higher in patients with diabetic neuropathy [( 12. 6 ± 3.6 ) μmol/ L] than without diabetic neuropathy [( 8. 2 ± 0. 9 ) μmol/L] ( P ＜0. 001 ), and the relationship remained significant after adjusting for duration of diabetes, glycosylated hemoglobin A1c (HbA1c), age, renal status, serum folate acid and vitamin B12, and metformin [OR 1.15( 1.02-1.28 ) ,P ＜ 0. 05]. In addition, per increase of 4. 0 μmol/L plasma homocysteine was closely related to the occurrence of neuropathy after controlling for per unit increase of other confounding factors [OR 1.17(0. 94-1.33), P ＜ 0. 05]. Conclusions Hyperhomocysteinemia was an independent risk factor for the occourence of diabetic peripheral neuropathy.