[Objective]To observe the bone repairing efficacy of large segmental femur defects in rabbits with calcium phosphate cement(CPC)combined with bone morphogenetic protein(BMP)and massive bone allograft,which may benefit the clinical application of large segmental bone allograft transplantation.[Method]Fifty-four New Zealand white rabbits were divided into three groups and a 2 cm femur defect was created on one side of each rabbit,followed by implantation with:CPC combined with BMP and massive bone allograft(Group A),bone allograft only(Group B)and autograft transplantation(Group C).Intramedullary nails with a 3 mm diameter fixed all the grafts transplanted.The bone defect repair efficacy was evaluated by radiology and histology exam at 4,8 and 12 weeks after operation.[Result]The bone reparation capacities of allograft with CPC/BMP complex was better than that of the allograft alone after 4 ~8 weeks of transplantation,which were similar to the result of autograft transplantation.Complete bone union was achieved for all the groups after 12 weeks of operation,with better bone remodeling for group A and group C transplantation.The healing process of CPC/BMP combined with allograft transplantation was featured with large amount of bone callus forming surrounding the graft-host bone union area and the surface of allograft,which composd the extra cortical bone bridge and ingrowth(EBBI).Bone invasion,resorption as well as new bone genesis were seen in the surface of bone allograft at early stage,companied by expansion of Haversians canal,with more lanner cells,osteoblasts,osteoclasts and blood cells inside the allograft.CPC was slowly biodegraded with the bone graft resorption and new bone regeneration.[Conclusion]CPC combined with BMP can improve the bone reparation and substitution process in massive bone allograft transplantation for the treatment of large segmental bone defects.

Objective: To explore antitumor responses induced by recombinant vaccinia viruses expressing a p53 antigenic peptide (rVV-p53M) and enhanced effect of recombinant vaccinia viruses expressing costimulatory molecule B7 (rVVB7). Methods: A 135 Cys to Tyr point mutation p53-transduced P815 mastocytoma (P815-mp53) was used as an experimental tumor and the p53 protein as the model of tumor associated antigen． rVV-p53M and rVV-B7 were used as vaccine to test their induction of CTL and antitumor immunity. Results: Immunization of BABL/c mice with rVV-p53M could elicit specific CTLs, which could specifically lyse P815-mp53 cells. Immunization of mice with rVV-p53M could survive a part of mice challenged with 1×106 P815-mp53. Treatment with rVV-p53M could significantly prolong the survival oftumor-bearing mice. Admixture at 1:1 ratio of rVV-p53 M and rVV-B7 could enhance antitumor responses induced by rVV-p53M. Conclusion: Immunization with recombinant vaceinia virus expressing antigenic peptide is a useful alternative to peptide-based vaccine. Costimulatory molecule B7 can enhance antigenic peptide to induce antitumor responses.

Penicillin expandase, also known as deacetoxycephalosporin C synthase (DAOCS), is an essential enzyme involved in cephalosporin C biosynthesis. To evaluate the catalytic behaviors of penicillin expandase under high penicillin G concentration and to identify mutants suitable for industrial applications, the specific activities of wild-type DAOCS and several mutants with increased activities toward penicillin G were determined by HPLC under high penicillin G concentrations. Their specific activity profiles were compared with theoretical predictions by different catalytic dynamics models. We evaluated the specific activities of wild-type DAOCS and previous reported high-activity mutants H4, H5, H6 and H7 at concentrations ranging from 5.6 to 500 mmol/L penicillin G. The specific activities of wild-type DAOCS and mutant H4 increased as penicillin G concentration increased, but decreased when concentrations of substrate go above 200 mmol/L. Other mutants H5, H6 and H7 showed more complex behaviors under high concentration of penicillin G. Among all tested enzymes, mutant H6 showed the highest activity when concentration of penicillin G is above 100 mmol/L. Our results revealed that the substrate inhibition to wild-type DAOCS' by penicillin G is noncompetitive. Other DAOCS mutants showed more complex trends in their specific activities at high concentration of penicillin G (>100 mmol/L), indicating more complex substrate inhibition mechanism might exist. The substrate inhibition and activity of DAOCS mutants at high penicillin G concentration provide important insight to help select proper mutants for industrial application.
Catalysis ; Intramolecular Transferases ; genetics ; Mutation ; Penicillin G ; pharmacology ; Penicillin-Binding Proteins ; genetics ; Streptomyces ; enzymology ; genetics

Objective To evaluate the safety and efficacy of 192Ir intraluminal brachytherapy for the prevention of urethral re-stricture after transurethral incision or transurethral resection of scar. Methods From Mar. 2004 to Jun. 2006,48 patients aging 18-81 years were treated by 192Ir intraluminal brachytherapy. The length of stricture(0.5-5.5 era) was≤3.0 cm in 90% of the patients. The stricture was caused by trauma in 23 patients and prostate hyperplasia operation in 19 patients. The cause of remaining 6 patients was unclear. All patients were diagnosed by urethra photograph or endoscopy. Radiotherapy was the initial treatment in 26 patients and the second time treatment in 22. The irradiation dose was from 14 Gy to 18 Gy.Results The median follow up was 10 months,and the total response rate was 98%. Only one patient recurred and received transurethral incision again. The uresis was fluency in 47 patients and the maximum flow rate was 13.9-36.4(19.2±10.3) ml/s. No secondary urethral bleeding or urethral cancer was observed.Conclusions Being a safe and feasible treatment, ,192Ir intraluminal brachytherapy following transurethral incision or transurethral resection of scar can effectively prevent urethral re-stricture.

BACKGROUND: Now, Shock Wave Therapy is used to cure the ununion and delayed union of bone, the avascular necrosis of the femoral head and the chronic injury of locomotor system, what has got a good curative effect. But, the mechanism is not clear. OBJECTIVE: To investigate the expression of c-Fos, c-Jun protein in human bone marrow mesenchymal stem cells (BMSCs) Influenced by shock wave.DESIGN, TIME AND SETTING: Randomized group, the controlled study was performed at the State Key Laboratory of Pathology, College of Basic Medicine, Jilin University, from March 2007 to March 2008. MATERIALS: Bone marrow was obtained from healthy volunteers.METHODS: Human BMSCs were cultured in vitro. And the fourth generation cells were digested into cell suspension with 2.5 g/L trypsin and adjusted at a density of 1.0×10~9/L with DMEM-LG. Then, the cells were divided into 6 1.5-mL Eerrendorf tubes, one was control group and the other five were experimental groups. The experimental groups were treated with an optimal dose of shock wave (8.5 kV, 120 times) by liquid-electric shock wave lithotripsy. Then the protein was extracted at different time points (5, 15, 30 minutes, 1, 2 hours) after treatment.MAIN OUTCOME MEASURES: Inverted microscope was used to observe the morphologic feature of BMSCs. The growth curves were charted by MTT method. The change of activations of c-Fos, c-Jun were tested by western blot. RESULTS: ①BMSCs were seeded in culture plate. The cells began to divide and proliferate slowly 24 hours later, and became fusiform shape after adhering to the wall. 3 days later, the speed of proliferation quickened, and cells accumulated colony. At day 10-14, the number of hMSCs grew till they covered the bottom of the culture plate. The round passage hMSCs adhered to the wall completely in 24 hours, which were similar to primary cells. The cells connected together during one week, and showed vortex-like. The speed of proliferation became slower and the cells became older when hMSCs were passaged to the tenth generation. ②MTT method showed that the growth curve of original, P2, P3 hMSCs looked like S shape, and the third to fifth days were growing period. ③The phosphorylation level of c-Fos and c-Jun began to increase after induced by shock wave and reached a peak at 30 and 15 minutes, respectively. And their phosphorylation level were 2.56-fold and 1.68-fold (P < 0.01) compared with the average level in control groups, respectively. Then they began to decrease. There was no apparent change in the total dose (P > 0.05). CONCLUSION: Following the treatment of shock wave, the activations of c-Fos and c-Jun in BMSCs increased.

Objective: To explore antitumor responses induced by recombinant vaccinia viruses expressing a p53 antigenic peptide (rVV p53 M) and enhanced effect of recombinant vaccinia viruses expressing costimulatory molecule B7 (rVV B7). Methods: A 135 Cys to Tyr point mutation p53 transduced P815 mastocytoma (P815 mp53) was used as an experimental tumor and the p53 protein as the model of tumor associated antigen. rVV p53 M and rVV B7 were used as vaccine to test their induction of CTL and antitumor immunity. Results: Immunization of BABL/c mice with rVV p53 M could elicit specific CTLs, which could specifically lyse P815 mp53 cells. Immunization of mice with rVV p53 M could survive a part of mice challenged with 1?10 6 P815 mp53. Treatment with rVV p53 M could significantly prolong the survival of tumor bearing mice. Admixture at 1∶1 ratio of rVV p53 M and rVV B7 could enhance antitumor responses induced by rVV p53 M. Conclusion: Immunization with recombinant vaccinia virus expressing antigenic peptide is a useful alternative to peptide based vaccine. Costimulatory molecule B7 can enhance antigenic peptide to induce antitumor responses.

OBJECTIVE To study the isolation,distributive characteristics and drug resistance of AmpC enzyme producing Gram-negative bacilli in nosocomial infection of two years and provide the evidence for treatment. METHODS A total of 528 strains of Gram-negative bacilli collected from daily specimens were identified with Bio-Fosun-Ⅰ,and AmpC enzyme was screened by cefoxitin disk and then corroborated by EDTA disk method. All data were analyzed statistically. RESULTS Among 528 strains collected,136 (25.75%) were AmpC enzyme producing strains,the respective percentage of Pseudomonas aerugionsa,Echerichia coli,Enterobacter cloacae,Klebsiella pneumoniae and Citrobacter was 32.35%,28.67%,18.38%,8.09% and 5.15%,respectively. Most strains (38.9%) were detected in ICU. The common infection sites were lungs. The resistance rate of AmpC enzyme producing strains to the first,second and third-generations cephalosporins was 71.3-99.5%. The susceptive rate of AmpC enzyme producing strains to imipenem,cefepine,amikacin and piperacillin/tazobactant were low. CONCLUSIONS For effective supervision and control of AmpC enzyme producing Gram-negative bacilli in nosocomial infection,detection of AmpC enzyme shoud be paid much attention by clinical microbiology laboratory.

BACKGROUND:Studies addressing coronary heart disease are largely dependent on the establishment of myocardial infarction animal models. It is very important that exploring a safe method with easy operation, less damage, long time survival and high survival rate for myocardial infarction animal model OBJECTIVE:To compare the pros and cons of two kinds of thoracotomy anterior descending coronary artery ligation to do myocardial infarction animal model. METHODS: Thirty healthy male New Zealand white rabbits were randomly divided into three groups: control, median sternotomy incision, and left sternal incision. The anterior descending coronary artery was ligated after thoracotomy. The operation time, amount of intraoperative bleeding, postoperative food intake, and recovery time of eating were monitored during the surgery and within 24 hours after the surgery. And myocardial enzyme indexes were also monitored within 24 hours after the surgery. Rabbits were detected with ultrasonic echocardiogram at 4 weeks. RESULTS AND CONCLUSION:Different levels of ST segment elevation appeared in median sternotomy and left sternal incision groups by echocardiogram. The success rate of modeling was 70% in median sternotomy incision group, and 80% in left sternal incision group. Within 24 hours post-surgery, the myocardial enzyme indexes in the two groups were significantly increased compared with before surgery (P < 0.05). At 4 weeks, the left ventricular ejection fraction and the left ventricular shortening fraction were significantly decreased when compared to before surgery (P< 0.05). The operation time was shorter, the amount of bleeding was less, the time of eating recovery was less and the amount of eating was much in median sternotomy group than in left sternal incision, with significant differences between he two groups (P < 0.05). The median sternotomy incision for the ligation of anterior descending coronary artery is better than the left sternal incision to establish myocardial infarction models.

Objective To investigate the relationship between monocyte chemoattractant protein-1 (MCP-1) A-2518G single nucleotide polymorphism (SNPs) and susceptibility of epithelial ovarian cancer(EOC) in Chinese Han population of Hunan region.Methods MCP-1 A-2518G SNPs of the EOC were genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis in 92 patients with EOC and 38 healthy women as control.Results MCP-1 A-2518G SNPs had AA,AG,and GG genotypes in cancer and control groups.The frequencies of AA,AG,and GG genotypes were not significantly different between cancer and control groups (AA:17.40％ and 15.79％ ; AG:44.56％ and 52.63％ ; and GG:38.04％ and 31.58％ ; P ＞0.05).Multivariate logistic regression analysis showed that there was no significant correlation between MCP-1 A-2518G polymorphism and EOC (P ＞0.05).Conclusions This present study suggested that MCP-1 A-2518G polymorphism should not be related to susceptibility of EOC in the Chinese Han population of Hunan region.

ObjectiveTo evaluate the diagnostic value of ultrasound or CT guided percutaneous Trucut needle biopsy on the diagnosis of pancreatic tumors.Methods One hundred and twenty-four patients clinically diagnosed as pancreatic cancer without pathological diagnosis underwent percutaneous pancreatic biopsy by using Trucut needle under ultrasound or CT guidance.ResultsOne hundred and nine procedures of ultrasound-guided biopsy and 15 procedures of CT-guided biopsy were performed,and one patient received 2.3times of punctures.Tissue samples were obtained in all 124 patients,the diagnostic accuracy was 95.2％,among them 115 were adenocarcinoma,5 were cystadenoma,2 were metastasis cancer,1 was cancer of unknown origin and 1 was normal.The sensitivity,specificity,and accuracy were 99.2％ 100％,and 99.2％,respectively.Transient serum amylase increase was observed in 3 patients; 5 patients' abdominal pain aggravated,but all recovered with conservative management.One patient was found to have tumor seeding on the spot of insertion after 34 days.No other major complications occurred.ConclusionsUltrasound or CTguided percutaneous pancreatic 16 ～ 18G Trucut needle biopsy is a safe and simple procedure with excellent diagnostic value for pancreatic cancer.