Objective To investigate the therapeutic efficacy of Video-assisted thoracoscopic surgery (VATS)in treating hemopneumothorax. Methods 83 cases of hemopneumothorax treated with exploratory thoracotomy with the help of VATS were retrospectively analyzed. Results All the 83 cases were completed successfully by the VATS.22 cases were treated with bunamiody resection,42 eases were treated with lung repair operation,12 cases were treated with pleuropexy,5 cases were treated with albumingel injection,2 cases were treated with thoracotomy because of serious injury.All patients were followed up 1 year and were found in good condition. Conclusion VATS could treat bunamiody resection and lung repair operation,can stop the bleeding effectively, cause less injury,good therapeutic effect and quickly recovery.

Objective To observe the diagnosis and surgical treatment of substemal goiter. Methods The clinical data of 26 cases with substemal goiter underwent thyroidresection were retrospectively analyzed. Results All the 26 cases were treated by operation. 20 cases were operated by Lower collar incision. 6 cases were operated by ster-notomy. The pathological results showed nodular goiter in 18 cases,follicular adenoma in 6 cases,and papillary carcinoma in 2 cases. Conclusions Surgery via cervical collar incision was fit for most of retrostemal thyroid nodules,and sternotomy or thoracotoray were safely fit for large or carcinomatous substemal goiter. Most cases of retrostemal goiter could be removed by Lower collar incision.

Objective:To explore the function of the cya gene and the preliminary mechanism of attenuated strain.Methods:The biological characteristics of cya mutant in acid and alkali resistant,salt resistance,motility,biofilm components,poisonous to the cells of epithelial cell adhesion,invasion were analysis.Results:The mobility capabilities,acid and alkali resistance and salt tolerance of cya mutant were significantly lower than the parent strain;the composition testing revealed that the cya mutant did not produce cellulose,curli and biofilm;at the same time the adhesion and invasion to epithelial cells of cya mutant had a prominent depression,and the toxicity to HeLa cells was weaker than the parent strain.Conclusion:The function of cya gene is closely related to athletic ability, penetration of cell membrane, the formation biofilm and virulence.It will provide a theory reference to the functional research of Salmonella typhimurium cya gene and the mechanism of attenuated strain.This will contribute to the development of oral vaccine using attenuated Salmonella typhimurium as vector.

In order to develop a safer vaccine strain exploit Salmonella Pullorum vaccine strain as a live vaccine vector.Methods:AΔcrpΔasd mutant of S.pullorum C79-13 strain was constructed and it was developed E.coli donor strain mutant was generated through the two-step method introduced by χ7213 ( pREΔasd) was conjugated with the recipient of C 79-13Δcrp.The C79-13ΔcrpΔasd the transduction of recombinant suicide plasmid .Results:PCR and sequencing results showed that ΔsipBSL1344 was suc-cessfully constructed.The further study indicated that foreign DAP must be supplied for the ΔcrpΔasd mutant to grow,in addition,the asd gene was transmitted stably .Compared with C79-13 strain,the O antigens was identical to C79-13 strain,but the growth velocity was reduced significantly ,significantly reduced virulence .Conclusion: The ΔcrpΔasd mutant can accept asd+plasmid to construct host-vector balance lethal system , and this system can be used to express foreign gene efficiently and to develop potential oral multivalent vaccines.

Objective:To discuss the effects of recombinant IFN-α-IL-18 fusion protein on chicken lymphocyte transformation and the chicken nuclear factor kappa B ( NF-κB ) activity.Methods: The recombinant plasmids pPICZ-IFN-α, pPICZ-IL-18 and pPICZ-IFN-α-IL-18 were constructed,and transformed into P.Pastoris X-33 strain by electroporation.The recombinant proteins were ex-pressed under the induction of 1% methanol,and detected by SDS-PAGE and Western blot.The biological activities of the expressed products were detected by the lymphocyte transformation test,the NF-κB concentration in chicken lymphocyte and lymphocytic nucleus were detected at different times with ELISA.Results:SDS-PAGE and Western blot showed that the expressed products existed in super-natant,and the molecular weights were about 22 kD,23 kD and 43 kD,respectively.The expressed products IL-18 and IFN-α-IL-18 could stimulate chicken lymphocyte transformation (P<0.05),the biological activity of IFN-αstimulating lymphocyte transform was feeble (P>0.05).Compared with the control group,the total NF-κB concentration in chicken lymphocyte induced by IL-18 and IFN-α-IL-18 were increased (P<0.05),and the NF-κB in lymphocytic nucleus was increased significantly (P<0.01).Otherwise,the total NF-κB in lymphocyte induced by IFN-αincrease was limited (P>0.05),but the NF-κB in lymphocytic nucleus showed the remarkable increase ( P<0.05 ) .Conclusion: IFN-α-IL-18 and IL-18 can promote lymphocyte transformation significantly, the activity of IFN-αinduced lymphocyte transformation is imperfect.The biological activity of stimulating lymphocyte transformation is associated with the NF-κB expression,activation and nucleo-cytoplasmic transport.The study built foundation for the function of IFN-α-IL-18 fusion protein and the exploration of the role of controlling epidemic diseases.

Objective:To discuss the effects of recombinant IFN-α-IL-18 fusion protein on chicken lymphocyte histamine induced and the NF-κB p65 activation and nucleo-cytoplasmic transport.Methods: The healthy chickens blood was sterile adopted with anticoagulant,then separation of the chicken peripheral blood lymphocyte and divided into 10 groups:The yeast expression and purification protein IFN-α-IL-18,IL-18,IFN-α were added with 250 ng/ml,500 ng/ml and 1 000 ng/ml respectively while the control was only added RPMI1640 with 3 repetitions for each group.Then the histidine decarboxylase activity,histamine,IFN-γ,PI3K,MAPK and NF-κB p65 in cell nucleus were detected.Results: The recombinant IFN-α-IL-18 and IL-18 could significantly promote the activity of histidine decarboxylase (P<0.01),increase the contents of histamine (P<0.01),induce IFN-γ (P<0.01),improve the contents of PI3K (P<0.01) and the NF-κB p65 levels in nucleus (P<0.01),and the higher concentration of IFN-α had a similar effect to lymphocytes.The effects of IFN-α-IL-18,IL-18 and IFN-α on MAPK was acratia.Conclusion: IFN-α-IL-18 and IL-18 can stimulate chicken peripheral blood lymphocyte populations increased histamine contents significantly and promote the induction of IFN-γ.IFN-α-IL-18,IL-18 and IFN-α increase PI3K expression in lymphocyte associated with the NF-κB activation and NF-κB p65 nucleo-cytoplasmic transport.The study built foundation for the function of IFN-α-IL-18 and the exploration of the mechanism of controlling epidemic diseases.

Objective:In order to develop an oral live vaccine vector of swine that can stably carry exogenous genes.Methods:Mutant ΔcrpΔcyaΔasdC78-1 was constructed by the method of suicide plasmid pREasd-mediated bacteria homologous recombination on the basis of attenuated Salmonella choleraesuisΔcrpΔcyaC78-1.Complementary plasmid pYA3493 with asd was electrotransformed into the mutant,and thenΔcrpΔcyaΔasdC78-1(pYA3493) host-vector balanced lethal system was constructed.Its biological characteristics were analyzed further.Results:The results of PCR and sequencing showed thatΔcrpΔcyaΔasdC78-1(pYA3493) was constructed suc-cessfully.Biological characteristics showed that the serotype of ΔcrpΔcyaΔasdC78-1(pYA3493) was identical to ΔcyaΔasdC78-1 and vaccine strain C500 and it can stably carry theΔasd gene in vitro;its growth speed was a little slower than ΔcrpΔcyaC78-1 strain,but both of their growth speeds were significantly slower than vaccine strain C500;the biochemical characteristics of ΔcrpΔcyaΔasdC78-1 ( pYA3493 ) were basically the same with ΔcrpΔcyaC78-1 strain.Oral virulence test in mice showed that the virulence ofΔcrpΔcyaΔasdC78-1 ( pYA3493 ) was similar with ΔcrpΔcyaC78-1, but its median lethal dose is 412 times of vaccine strain C500.Conclusion:These results demonstrated that attenuated Salmonella choleraesuisΔcrpΔcyaΔasdC78-1(pYA3493) strain had the potential to be used as an oral live vaccine vector for expressing foreign genes efficiently.

ObjectiveTo explore the feasibility and efficacy of percutaneous transhepatic approach in the treatment of intrahepatic and extrahepatic bile duct stones. MethodsNineteen patients with bile duct stones who were treated in our hospital from January 2000 to October 2014 were enrolled as subjects. In the treatment of bile duct stones, all patients received percutaneous transhepatic approach combined with the application of cholangioscopy and lithotripsy equipment. ResultsIn the 19 patients, 12 had the stones successfully removed in a single session, 5 had the stones completely removed through the liver sinus in three sessions, and 2 had residual stones in intrahepatic tertiary bile ducts. The success rate of lithotomy reached 89.47%(17/19). ConclusionThe percutaneous transhepatic approach achieves minor surgical invasion and slight pain in the treatment of bile duct stones, which can be selectively used based on patients′ condition.

Objective To analyze the biological characteristics of a mutant strain of Salmonella ty-phimurium SL1344 with sseK2-deletion (SL1344△sseK2) in order to provide reference for further study of safe and effective live vaccines. Methods The mutant strain SL1344△sseK2 with a deletion of 1047 bp in sseK2 gene was constructed through a two-step allelic exchange using recombinant suicide plasmid. Its com-plemented strain, SL1344C△sseK2, was also constructed. Biological and immunological characteristics of the mutant strain were detected. Results PCR, double-enzyme digestion and sequencing analysis showed that the mutant strain SL1344△sseK2 and the complemented strain SL1344C△sseK2 were successfully con-structed. The serotype of the mutant strain was 1,4,[5],12:i:1,2, identical to the parent strain SL1344. In addition, the mutant strain showed no significant change in biochemical characteristics or growth rate and was genetically stable in vitro. Compared with the parent strain SL1344, the virulence of SL1344△sseK2 was attenuated in BALB/ c mice. The median lethal dose of SL1344△sseK2 for 6-week-old BALB/ c mice was 3. 44×108 colony-forming units (CFU), which was 1620 times lower than that of SL1344. Oral immuniza-tion with SL1344△sseK2 protected 62. 5％ of the mice against challenge with wild Salmonella typhimurium strains on 17 d after vaccination. The levels of serum IgG antibody peaked on 14 d after immunization. No significant difference in biological characteristics was observed between the complemented and the parent strains, indicating that the mutant strain was basically complemented to the wild-type strain.Conclusions The mutant strain SL1344△sseK2 was constructed successfully and genetically stable with sig-nificantly attenuated virulence and good immunogenicity. This study suggested that sseK2 gene played an im-portant role in regulating the virulence of SL1344, which might provide reference for further study of its func-tion and for assessing its potential as a candidate live attenuated vaccine.

In order to research immunogenicity of the recombinant rVP2-IL-2 fusion protein, we obtained the rVP2-IL-2 fusion protein using Pichia pastoris expression system, and then evaluated its potential to induce immune responses in chicken. The effect was determined in the form of protective anti-IBDV VP2 titers, antibodies (IgG1 and IgG2a), lymphocyte proliferation, the levels of interferon-gamma and interleukin-4 cytokines, and challenge experiment. Antibody titers and proliferation lymphocyte level suggested that the fusion protein could elicit specific humoral immune and cellular immune responses, antibody sub-type results indicated that the rVP2-IL-2 fusion protein induced secretion both of IgG1 and IgG2a. The seem result elicited from cytokines ELISA test, secretion of both of Th1 (gamma-IFN) and Th2 (IL-4) were induced by the rVP2-IL-2 fusion protein. Challenge experiment result shown that chicken immunized the rVP2-IL-2 fusion protein obtained 85% protection. These results confirm that the fusion protein enhances the protection against IBDV through both humoral and cell-mediated immunity, and thus could serve as a candidate for the development of IBDV subunit vaccine.
Animals ; Antibodies, Viral ; biosynthesis ; blood ; Chickens ; immunology ; Immunoglobulin G ; blood ; Interleukin-2 ; biosynthesis ; genetics ; Pichia ; genetics ; metabolism ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology ; Th1 Cells ; immunology ; Th2 Cells ; immunology ; Vaccines, Subunit ; immunology ; Viral Structural Proteins ; biosynthesis ; genetics ; Viral Vaccines ; immunology