BACKGROUND: Methyl cobalamin, a derivative of vitamin B12, .can promote the metabolism of nucleic acid protein, axoplasmic transport and axonal regeneration.OBJECTIVE: To observe the role of target muscular injection of methyl cobalamin ofdifferent dosages in the regeneration of peripheral nerve injury in rats.DESIGN: A randomized and controlled animal experiment.SETTING: Changzhou Second People's Hospital affiliated to Nanjing Medical University.MATERIALS: The experiment was carried out in March 2003. Thirty healthy Wistar rats were randomly divided into three groups with 10 rats in each group: high-dosage group, low-dosage group and blank control group.METHODS: All the rats were made into models of sciatic nerve injury by epineural suture immediately after transection of left sciatic nerve. Rats in high- and low- dosage groups were injected with methyl cobalamin of 300and 100 μg/kg per day respectively, and those in blank control group were injected with 1 mL normal saline of the same volume, and target muscular injection was given once a day postoperatively. At postoperative 4 and 8weeks, 5 rats were killed at one time; the wet mass of left triceps surae muscle was measured. Morphological observation of sciatic nerve under light and electron microscopes and imaging analysis were made as the detected indexes to analyze the effect of methyl cobalamin of different dosages on sciatic nerve injury in rats.sciatic nerve myelin sheath at postoperative week 8.parison of the wet mass of triceps surae muscle at postoperative 4 weeks: It was higher in high-dosage group than in low-dosage group and blank control group [(1.367±0.012) g, (1.164±0.011) g, (0.950±0.009) g, P ＜ 0.05].week 8: It washigher in high-dosage group than in low-dosage group and blank control group [(2.205±0.015), (1.611±0.013), (1.230±0.014) g, P ＜ 0.05].weeks postoperatively: It was obviously higher in high-dosage group than in low-dosage group and blank control group [(13.30±1.167), (9.453±1.233),atic nerve myelin sheath at 8 weeks postoperatively: It was thicker in highdosage group than in low-dosage group and blank control group [(1.145±0.108),(0.806±0.065), (0.560±0.045) mm, P ＜ 0.05].CONCLUSION: Target muscular injection of methyl cobalamin can promote the regeneration of peripheral nerve, and methyl cobalamin of high dosage can better play its role in nourishing injured nerve.

Objective To examine the effect of target muscle injection with different dosages of Methycobal in the sciatic nerve regeneration of adult rats so as to determine a better method and proper dosage of Methycobal for clinical treatment of peripheral nerve injury. Methods A total of 30 healthy Wistar rats were employed to make sciatic nerve injury model through immediate suture of the adventitia posterior to transection of the left sciatic nerve. The rats were divided randomly into three groups, ie, high dosage group (Methycobal for 300 ?g/kg, Group A), low dosage group (Methycobal for 100 ?g/kg, Group B) and control group (1 ml normal saline once a day, Group C),10 rats in each group. At 4 and 8 weeks, 5 rats from each group were taken to observe the effect of different dosages of Methycobal on rat sciatic nerve regeneration, where the detection indices included morphological changes and image analysis of sciatic nerves by optic and electron microscopic investigations as well as tricipital muscle wet weight. Results Four weeks after operation, tricipital muscle wet weight was (1.367?0.012) g in the Group A, (1.164? 0.011) gin the Group B and (0.950?0.009) g in the Group C, with significant difference among groups A, B and C (P

Objective To introduce a sort of method about thumb reconstruction to complex thumb defect. Methods From January 2003 to December 2006, 13 patients who incur sever thumb defect above5 grade adopt the upper limb lateral bone-skin flap combined with the second toe transplant primary thumb reconstruction, pestop follow-up visit 12 months to 3 years, according to hand surgery society of Chinese Medical Associaition thumb and finger reconstruction functional assessment probation standard to evaluatethe function of reconstituted thumb. Results All of the transplanted upper limb lateral bone-skin flaps and the second toes take, and function well. Conclusion The upper limb lateral bone-skin flap combined with the second toe transplant primary repair complex thumb defect had been tested one good method for thumb reconstruction to above 5 grade thumb defect.

Objective To evaluated the feasibility and accuracy of obtaining the myocardial perfusion image and coronary artery image of pig models of acute myocardial infarction by the second generation dual-source CT dual-energy myocardial perfusion one-stop scanning. Methods The pig models of acute myocardial infarction was established by using the interventional gelatin sponge embolism method on 5 pigs. The myocardial perfusion scanning and coronary angiography were respectively used 20 min before and 24-48 h after modeling. The heart were taken out immediately after the inspection for multiple punch biopsy and HE staining, then the heart was sliced for triphenyltetrazolium chloride (TTC) staining. According to the gold standard of coronary angiography and pathological staining, the sensibility and speciifcity of the coronary artery image of acute myocardial infarction of pig reconstructed by myocardial perfusion scanning and the myocardial perfusion image were respectively evaluated, and the consistency of the results was veriifed by Kappa test. Results Compared with the pathologic gold standard, the sensitivity, speciifcity, of myocardial perfusion iodine image obtained by myocardial perfusion scanning were 93%, 91%; and Kappa value was 0.68. Compared with the coronary angiography gold standard, the sensitivity and speciifcity of coronary artery image obtained by myocardial perfusion scanning were 93%, 81%, and Kappa value was 0.71. Conclusions The second generation dual-source CT dual-energy myocardial perfusionone-stopscanning can accurately get the coronary artery and myocardial perfusion images of the pig models of acute myocardial infarction.

Objective To investigate the anaesthetic effect of propofol combined with remifentanil by target-controlled infusion (TCI) used in gynecological laparoscopic myomectomy.Methods Fifty cases,who were scheduled for gynecological laparoscopic myomectomy in our hospital from June 2010 to June 2011,was randomly divided into propofol combined with remifentanil group (n =25) and inhalation anesthesia (remifentanil combined with sevoflurane) group (n =25).The heart rate and blood pressure were recorded at the time of before induction of anesthesia (T0),30 min after carbon dioxide pneumoperitoneum,the end of operation and 3 min after extubation.The awake time,time of extubation and surgery time were also recorded.Results The hemodynamics were kept stable in propofol combined with remifentanil group,and there were no significant difference with respect to SABP,DABP and heart rate at all time points compared with baseline (P ＞0.05) in propofol group.However,in inhalation anesthesia group,SABP,DABP and heart rate were increased significantly at 30 min after carbon dioxide pneumoperitoneum and 3 min after extubation when compared with baseline (P ＜ 0.05) and were higher than those of propofol group (P ＜ 0.05) at counterpart time points.In inhalation anesthesia group,the awake time ((9.3 ± 1.5) min vs (4.9 ± 1.1) min,t =10.56,P =0.017) and time of extubation ((12.9 ± 2.4) min vs.(6.8 ± 1.2) min,t =12.36,P =0.01) were significantlv longer than that of propofol group (P ＜ 0.05).Conclusion Propofol combined with remifentanil TCI-based anesthesia could achieve the optimal hemdynamic stability during anesthesia maintance and better recovery profile from anesthesia in gynecological laparoscopic myomectomy.

Objective To test the protective immunity in mice induced by recombinant Schistosoma japonicum Sj14FABP through several adjuvant formulations. Methods The recombinant Schistosoma japonicum Sj14FABP was prepared by expression in E. coli as a GST fusion protein (rSj14/GST) and used to vaccinate outbred Kunming mice by using complete Freund's adjuvant (FCA)/incomplete Freund's adjuvant (FIA), Bacillus Calmette-Guerin (BCG) and the immunostimulating complex (ISCOM) as adjuvant respectively. Results The purified recombinant protein rSj14/GST was immunogenic in mice, and 34.3% and 36.0% worm reduction rates were obtained in outbred Kunming mice immunized intradermally with BCG adjuvant and immunized subcutaneously with ISCOM adjuvant respectively, compared with non-vaccinated control group. However, intramuscularly vaccination with rSj14/GST in FCA/FIA was not protective, although the high level of IgG antibody was induced. Conclusion Both BCG and ISCOM are suitable adjuvants for rSj14/GST.

Objective To investigate the pharmacokinetics and relative bioavailability of praziquantel injection in buffaloes in contrast to praziquantel tablet. Methods A single oral administration of praziquantel tablet at a dose of 20 mg/kg or intramus-cular administration of praziquantel injection at a dose of 10 mg/kg was performed in six healthy adult buffalos according to a two-period crossover design. The praziquantel concentration in plasma was determined by a high performance liquid chromatography (HPLC)method. The pharmacokinetic parameters were calculated by non-compartmental analysis. Results The main pharma-cokinetic parameters of praziquantel tablet were as follows:Tmax=(0.60±0.29)h,Cmax=(0.57±0.37)μg/ml,T1/2β=(0.70±0.42) h,AUC=(0.80±0.70)(μg/ml)·h. The main pharmacokinetic parameters of praziquantel injection were as follows:Tmax=(0.65± 0.49)h,Cmax=(3.82 ± 1.17)μg/ml,T1/2β=(1.00 ± 0.73)h,AUC=(1.61 ± 0.89)(μg/ml)·h. The relative bioavailability of pra-ziquantel injection was 402.5％in contrast to praziquantel tablet. Conclusion The praziquantel injection has pharmacokinetic characteristics of rapid absorption,high bioavailability and extensive distribution,and the clinical recommended dosage of pra-ziquantel injection is 10 mg/kg.

Objective To express the gene encoding mature protease of Schistosoma japonicum asparaginyl endopeptidase (Sj32) and evaluate the potential of the recombinant protein rSj32 in diagnosis of domestic animal schistosomiasis. Methods The DNA fragment encoding mature protease of Schistosoma japonicum asparaginyl endopeptidase was cloned with PCR from pET-28(a)/Sj32, and a recombinant plasmid was previously constructed in the laboratory, which contained the ORF of the gene encoding the pro-enzyme Sj32. The amplified DNA fragment was subcloned into pET-28a( + ) and the recombinant plasmid was transformed into E. coli BL21 (DE3) to express the mature protease of Sj32. Then the recombinant antigen (rSj32) was used in ELISA assay to diagnose schistosomiasis of mice, rabbits and water buffalo artificially infected. The detection effects of soluble Schistosoma japonicum egg antigen (SEA) , rSj32 and the recombinant 23 KDa membrane protein were compared. Results The recombinant antigen rSj32 with a molecular weight 41 KDa was successfully produced in E. coli BL21 ( DE3) and was purified with His Column with a yield of 25 mg/L E. coli culture. By using rSj32 as coating antigen in ELISA assay to detect the specific antibody in artificially infected mice, rabbits and buffalo, the sensitivities were 88.9% , 85.0% and 71.8% , respectively, the specificities were 100% , 96.7% and 96. 9% , respectively. There were no significant differences among the detection results of rSj32, SEA and rSj23. Conclusion rSj32 is a promising antigen for serological diagnosis of domestic animal schistosomiasis.

Objective To screen protective antigen genes and construct the T7 phage display library from adult worms of Schistosoma japonicum.Methods Total RNA was extracted from adult worms of S.japonicum by Trizol reagent and mRNA was isolated from the total RNA.The ds cDNA was synthesized by reverse transcription using random primer.Directional EcoRⅠ/HindⅢ linkers were ligated into the ends of ds cDNA and the ds cDNA was digested with EcoRⅠand HindⅢ,which resulted in ds cDNA with EcoRⅠand HindⅢ adhering ends.The digested ds cDNA fragments longer than 300 bp in length were fractionated and ligated into T7 Select 10-3b vector.After packaging in vitro,the T7 Select 10-3b vector was transformed into BLT5403 to construct the T7 phage display cDNA library.Plaque assay and PCR were used to evaluate the library.Seven known objective genes of S.japonicum were screened by PCR to detect the representation of the library.Result Primary library capacity was 4.98?106 pfu,and the titer of amplified library was 3.85?1011 pfu/mL.The PCR identification result of 96 clones picked at random showed that recombination rate was 93.8%,in which 95.6% inserted cDNA fragments were longer than 300 bp in length.All the seven known objective genes of S.japonicum were amplified from the library.Conclusion The T7 phage display library from adult worms of Schistosoma japonicum was constructed.

Objective To look for the genes of Schistosoma japonicum related to the Schistosoma-resistance of Microtus fortis.Methods The fresh sera of Microtus fortis were used to screen a T7 phage display cDNA library from worms of Schistosoma japonicum established in our lab.The positive clones were sequenced and functionally analysed through bioinformatics.Results The specific phages binding to the sera of Microtus fortis were enriched 857-fold after three rounds of biopanning,and 58 positive clones picked at random were sequenced and 10 ESTs were obtained.BLASTn results showed that 7 ESTs had 99%-100% similarity to the genes of Shistosoma japonicum reported in GenBank and 1 EST had 82% similarity to a zinc finger protein encoden gene from Pan troglodytes.The results of these ESTs function prediction indicated most of them were involved in the regulation of gene expresion of Schistosoma japonicum.Conclusions Several target genes of Schistosoma japonicum related to the Schistosoma-resistance of Microtus fortis are obtained and those would lay foundation to expatiate the native resistance mechnism of Microtus fortis to Schistosoma japonicum.