Objective To study the expression of basic fibroblast growth factor (bFGF) in ectopic osteogenesis of autogenetic minimal morselized bone so as to discuss the bone formation of minimal morselized bone. Methods All 48 rabbits were divided into two groups randomly. Then autogenetic minimal morselized bone and bulk bone were implanted into the muscle bag models of gluteus maximus muscle respectively. Samples were harvested on day 1,3,5,7,11,14,21 and 28 postoperatively and tested by the methods of histology, immunohistochemistry (IHC) and in situ hybridization (ISH). Results (1) The morselized bone grew faster than the bulk bone and was replaced by neonatal bone on the 28th day. In the group of bulk bone, the ability of osteogenesis was weaker dominated by bone absorption. (2) In the morselized bone group, the expression peaks of bFGF and bFGFmRNA appeared at day 5-7 postoperatively, mainly appeared in the mesenchymal cells, fibroblast, chondrocyte and osteoblast by the method of IHC and ISH. While in the group of bulk bone, the expressions of bFGF and bFGFmRNA were similar to those in the morselized bone group. The difference between the two groups was significant ( P

Objective To study the efficacy of repairing bone defect with the composite of allogeneic morselized bone, bone morphogenetic protein(BMP)and collagen. Methods 1.5 cm long bone defect was created in each radius of 34 New Zealand rabbits. A composite of 200mg allogeneic minute morselized bone with 10 mg BMP and 0.2 ml collagen was placed into the bone defect of left radius in each rabbit (group A); the contralateral bone defect was filled with a composite of 200 mg allogeneic morselized bone and 0.2 ml collagen (group B). The bone mineral density(BMD)of the defects was evaluated at 8 and 12 weeks. The biomechanic tests were examined at 12 weeks. In group C, only 0.2 ml collagen was implanted as control. Results Radiological and histological examination revealed that the defects healed completely in both experimental groups, but there were more new bone formation, early bridging and rapid healing in group A compared with that of group B. The value of BMD in group A was higher than that in group B. The three-points bending test of group A showed that ultimate strength and bending stiffness were superior than that of group B, the anti- axial compressive stiffness as well; and the torsion rigidity and maximal torque of group A were also greater than that of group B. There were significantly statistic differences in all biomechanical tests between the two groups. Histological evaluations displayed that there were significant differences in matured level of new bone formation and remodeling process between two experimental groups. The new lamellar cortical bone formation was more matured, and the remodeling process was more perfect in group A comparing to that of group B, which might contribute to the better mechanical properties of group A. The defects did'nt achieve any osseous healing in control group. Conclusion The allogeneic morselized bone can be used effectively to repair segmental bone defects. The experiment suggests that the bone healing process can be promoted by the composite morselized bone graft with BMP and collagen.

Objective To explore the possible mechanism of repetitive transcranial magnetic stimulation (rTMS) in learning and memory ability of Alzheimer's disease (AD) rats. Methods 90 SD rats with no differ-ence in learning and memory were selected and 30 rats were randomly selected as control group. The rest 60 were used for establishment of AD model by injecting A beta 1-42 into bilateral hippocampus of rats ,which was verified by Morris water maze test and immunohistochemistry. The successfully prepared model rats were randomly divided into AD group and (rTMS+AD) group,30 rats in each group. The (rTMS+AD) group was treated with rTMS,and the control group and AD group with pseudo stimulation. After intervention,Water maze test,EEG acquisition and analysis,immunohistochemistry and statistical analysis were done. Results (1)Compared with control group, the escape latency of AD group was longer and the average distance from the platform was farther away. Compared with AD group,the escape latency of (rTMS+AD) group was significantly longer and the average distance from the platform was significantly farther away (P<0.05).(2)Compared with control group,the LZ complexity of EEG signals in AD group was decreased. Compared with the AD group,the LZ complexity of EEG signals in (rTMS+AD) group was significantly increased statistically(P< 0.05).(3)The results of immunohistochemistry showed that there was no plaque in the brain tissues of control rats. The morphology of the cells was clear ,and the cells arranged neatly. Myelin staining was deeper and the distribution was long andconnected. There was plaque in the brain tissues of AD rats. Brain tissue cells became atrophy ,the morphology of the cells was not clear ,and the cells arranged randomly. Myelin staining was lighter and the distribution was short and disconnected. Compared with AD group,the cell atrophy of (rTMS+AD) group was reduced,and Myelin staining became deeper. Conclusions rTMS can increase the distribution of myelin in brain tissue ,raise the LZ complexity of EEG signals in AD model rats , and improve the learning and memory ability of them.

Aim To establish a HPLC method for de-termining cimifugin in rat plasma and investigate the pharmacokinetic characteristics of cimifugin in rats. Methods The plasma concentration of cimifugin was detected by HPLC in acetonitrile protein precipitation method after intragastric administration of cimifugin. The pharmacokinetic parameters were calculated by the procedure of DAS 2 . 1 . Results The regression equa-tion of cimifugin in rats plasma was Y =0. 187 X -0. 0236 (R2 =0. 998 2),which shows a good linear re-lation at 1 - 70 mg · L-1 . The concentration-time curves conformed to two-compartment model. The main pharmacokinetic parameters of Tmax, Cmax, T1/2α, T1/2z, Vd ,AUC(0-t) and AUC(0-∞) were 80 min, 10. 359 mg ·L-1 , 93. 131 min, 2. 179 L · kg-1 , 1946. 085 mg ·L-1 · min, 2138. 57 mg · L-1 · min, respectively. Conclusions We established a HPLC method to de-termine the concentration of cimifugin in plasma. The method is so highly specified and sensitive that it can be used in quantitative analysis in vivo on cimifugin. Cimifugin can be rapidly absorbed, reach the highest concentration and produce effect.

Objective This study was to investigate the value of CT guided 125iodine implantation combining transcatheter arterial chemoembolization(TACE) to prevent digestive tract bleeding in patients with portal vein tumor thrombus in primary hepatocellular carcinoma. Methods Forty patients with portal vein tumor thrombus which were diagnosed to have primary hepatocellular carcinomas by diagnostic criteria of Chinese Anti-Cancer Association were collected prospectively. They were divided into the treatment group and the control group, with 20 patients in each group. The treatment group was treated by TACE for hepatic tumor and 125iodine seed implantation for portal vein tumor thrombus, while the control group was treated by TACE for hepatic tumor and only given β-blockers medicines after treatment. Intraoperative and postoperative surgery-related complications were observed. Three months after surgery, enhanced abdominal CT scanning was performed to evaluate treatment effects which were divided into complete response (CR), partial response (PR), and progressive disease (PD) and stable of disease (SD), and the local tumor control rates were calculated. The bleeding rates and mortality after 3 months, 6 months, 12 months were recorded. Treatment effects of the two groups were compared with continuously correction Chi-square test, bleeding rates were compared with Fisher test, and survival rates were analyzed with Kaplan-Meier survival curve and compared with Log-rank test. Results Overall the 40 patients were treated successfully without serious surgery-related complications. In the treatment group, there were 8 patients with PR, 6 with SD and 6 with PD, and the local control rates were 40% (8/20). In the control group, there were 1 patient with PR, 6 patients with SD and 13 with PD. The difference of the local control rates was statistically significant (χ2=5.161, P=0.023).The bleeding rates at 3, 6 and 12 months were 2, 2 and 3 cases in the treatment group, for control group they were 2, 6 and 10 cases respectively. There was no statistical difference between the 3 months and 6 months bleeding rates (P values were 1.000 and 0.235), but for 12 months bleeding rates, the difference was statistically significant (P=0.041).The 1 year cumulative survival rates of the treatment group and control group were 70% (14/20) and 40% (8/20), and the difference was statistically significant (χ2=4.675, P=0.031). Conclusion The treatment of 125iodine implantation combining transcatheter arterial chemoembolization in patients with portal vein tumor thrombus in primary hepatocellular carcinoma can reduce variceal bleeding rate and improve survival rate.

Objective:To artificially design and express a recombinant protein named as ScFv-pLLO by fusing ScFv gene of Rituximab(C2B8)and dominant antigen epitopes from listeriolysin O(LLO),and studying its anti-tumor activity.Methods:VH and VL gene sequences of C2B8 against CD20 were acquired by searching United States Patent database,and ScFv sequence was constructed by linking VL and VH with a short peptide linker.Two CD4+T cell epitopes from LLO were selected and designed to splice ScFv sequence.The recombinant gene of ScFv-pLLO was cloned into prokaryotic expression vector and purified after induction.The capacity of ScFv-pLLO target-binding to B-cell lymphomas was evaluated by flow cytometry ( FCM ) and co-immunoprecipitation ( Co-IP ) .The effects of ScFv-pLLO on B-cell lymphomas proliferation and apoptosis were detected respectively.The immunogenicity of ScFv-pLLO was assessed by lymphocyte proliferation assay.Results: ScFv-pLLO was successfully expressed.It could bind to different B-cell lymphomas cell lines and obviously inhibit the growth of Raji cells as well as inducing apoptosis.Moreover,ScFv-pLLO was able to stimulate proliferation of spleen lymphocytes of immunized mice.Conclusion: The recombinant protein ScFv-pLLO can target-bind to B-cell lymphomas,and perform inhibitory effect and induce apoptosis on Raji cells that indicate ScFv-pLLO retain the capacity of ScFv derived from monoclonal antibody against CD20.Besides, ScFv-pLLO can induce immune response.This study provides a basis for further research about the role of ScFv-pLLO on simulating tumor cell antigens as well as being tumor vaccine adjuvant.

To evaluate the antiosteoporotic effects of benzyl benzoate glucosides from Curculigo orchioides (COBG) in ovariectomized (OVX) rats.

OBJECTIVETo study optimal process conditions for the inclusion of volatile oil from Nardostachyos Radix et Rhizoma using saturated water solution method.

METHODThe optimal technology of inclusion was identified respectively by the utilization ratio of volatile oils and the yield of inclusion compound in an orthogonal experiment design, and the inclusion efficiency was evaluated by TLC, UV, IR, SE and GC-MS.

RESULTThe optimum preparation conditions for inclusion were established as follows: volatile oil: beta-CD was 1: 6, the inclusion temperature was 40 degrees C, the stirring speed and time were in 300 r x min(-1) and for 1 h. The analysis results showed that a new substance was conformed. The change of the volatile oil component was found before and after inclusion.

CONCLUSIONThe optimal process conditions are easy for operation, and high in utilization ratio of volatile oils and the yield of inclusion compound.

Fibro-optic bronchoscopy (FB) examinations were undertaken in 42 cases with lung cancer and 25 cases with benign lung disease; methylation-specific PCR was performed in plasma, sputum and bronchoalveolar lavage fluid (BALF) specimen for detection of p16 gene promoter methylation in all patients.Of the 42 cases of lung cancer, the positive rates of p16 gene promoter methylation were 59.5% in BALF, 52.4% in plasma and 47.6% in sputum, respectively; while p16 gene promoter methylation was detected only in one plasma sample from 25 cases with benign lung disease ( P ＜ 0.05 ).The sensitivity,specificity and overall accuracy of FB were 59.5%, 100.0% and 74.6%, respectively.The sensitivity,specificity and overall accuracy of FB combined with aberrant p16 gene methylation in diagnosis of lung cancer were 92.9%, 96.2% and 94.0%, respectively.The FB examinations combined with detection of aberrant p16 gene methylation can further improve the accuracy to diagnosis of lung cancer.

Aim To explore the expressed level of ini-tiative key factors TSLP and IL-33 in a human kerati-nocyte cell line,HaCaT cells were chosen to be stimu-lated by different stimulants,and develop a stable and effective in vitro model to observe allergic sensitization. Methods HaCaT cells were cultured in K-SFM with different stimulants to screen out the stimulants which could significantly improve the expressed level of TSLP and IL-33.Expressed level of TSLP and IL-33 was an-alyzed by ELISA kits and immunofluorescence.Re-sults (1 )The dose-response relationship of single stimulant indicated that both Poly(I:C)and TNF-αcould significantly improve expressed level of TSLP and IL-33 in HaCaT cells,but the rest of stimulants was not observed significant stimulation in concentration range of this experiment.(2)Dose-effect relationship of combined stimulants indicated that poly(I ∶C)1 00 mg·L -1 combined with TNF-α20 μg·L -1 was the most efficient.(3)Time-effect relationship of the a-bove-mentioned combined stimulants showed that 1 2 h was the optimal time of stimulation.Conclusions Different stimulants and different time result in various expressed levels of TSLP and IL-33 in HaCaT cells.1 2 h stimulus duration of Poly(I:C)1 00 mg·L -1 com-bined with TNF-α20 μg · L -1 is the most efficient stimulating way.This result provides an effective in vitro model to study the pathomechanism and drug effi-cacy of allergic sensitization.