This study investigated the protection against the ND in chickens by a recombinant DNA vaccine. A plasmid vector encoding NDV F protein, which is reqired for virus cell fusion and is important for vaccine induced immunity, was used as a model to study how DNA vaccines may be modulated by the simulaneous expression of chicken IL-2. The NDV D26 strain F gene with CMV promotor and BGH polyA signal sequence was amplified by PCR from eukaryotic plasmid pcDNA-F, which contains the full-length NDV F gene, and clond into reconstructed eukaryotic plasmid pcDNA-IL2, which contains chicken IL-2 gene. Restriction endonuclease cleavage and PCR amplification showed that a bicistronic plasmid encoding NDV F gene and chicken IL-2 separately was successfully constructed. Two-week-old SPF chickens were intramuscularly innoculated the recombinant plasmid. Antibody and lymphocyte proliferative assay showed that the humoral and cellular immunity of chickens vaccinated the recombinant plasmid greatly increased compared with those innoculated only plasmid expressing NDV F protein. Challenged with the lethal dose of NDV F48E9 strain, 72% chickens vaccinated recombinant plasmid were survived, and 30% chickens vaccinated plasmid expressing F protein were survived. These results proved the adjuvant effect of chicken IL-2, and further showed that the efficacy of a DNA vaccine can be greatly improved by simultaneous expression of IL-2.

Objective To evaluate the efficacy of preoperative aspects and dimensions used for an anatomical (PADUA) scores in determining the surgical approach for T1 stage renal masses.Methods From Jan 2010 to Dec 2012,clinical data of 122 cases (76 males and 46 females),who underwent surgery for T1 stage renal masses,were collected retrospectively.The mean age was 51 years(range 21-81) and mean body mass index was (22.8±3.9) kg/m2.Sixty-three tumors were found in left kidney and 59 in right kidney.Among them,78 patients were diagnosed as T1a stage and 44 patients were T1b stage.In patients with T1a stage,56 received nephron sparing surgery (NSS) and 22 received radical nephrectomy (RN).In patients with T1b stage,21 received NSS and 23 received RN.The PADUA nephrometry score was analyzed to evaluate their relationships to surgical type and the approach of NSS.Results According to the PADUA nephrometry score,the number of low risk,middle risk and high risk patients were 24,62,26,respectively.Inlow risk group,middle risk group and high risk group,the proportion of RN and NSS was 8.3％/ 91.7％,30.6％/69.4％,66.7％/33.3％.In 77 patients received NSS,the unmber of laparoscopic NSS and open NSS was 18 ∶ 4,25 ∶ 18,2 ∶ 10,respectively.The PADUA nephrometry score was significantly associated with the type of surgery (x2 =23.16,P＜0.01),and the NSS approach (x2 =13.57,P＜0.01).Tumor size (HR =2.79 ; 95％ CI,1.29-6.02 ; P＜ 0.01),percentage of tumor deepening into the kidney (HR =3.82; 95％CI,1.77-8.09; P＜0.01),longitudinal (HR=4.00;95％CI,1.83-8.72; P＜0.01),tumor relationships with renal sinus(HR=103.13; 95％CI,21.85-486.81 ; P＜0.01),tumor relationships with urinary collecting system (HR =15.11 ; 95％ CI,5.95-38.35 ; P＜ 0.01),rim tumor location (HR =3.50 ; 95％ CI,1.61-7.59; P＜0.01) were closely related with surgery approach.The correlation coefficients of relationship with renal sinus was highest (r=0.70).Conclusions The PADUA nephrometry score provides a simple,useful and stable system to character the salient renal anatomy and guide the surgery.Low risk group should consider the NSS as the first line therapy.NSS could also be chosen in the middle risk group.However,the renal anatomy in those patients should be referred.RN should be chosen in high risk group.

Objective: To investigate whether cancer-associated- fibroblasts (CAF), the key component of tumor microenvironment, regulate the chemoresistant capacity of lung cancer cell line A549 through SDF-1 secretion. Methods: Primary cell isolation techniques was used to isolate cancer-associated-fibroblasts from lung cancer patients. MTT assay was applied to determine the proliferation and chemoresistance of A549 cells. Quantative PCR was used to detect the mRNA changes of Bcl-xL. Western blotting was used to detect the protein expression of Bcl-xL. ELISA was applied to detect the SDF-1 secretion from normal fibroblasts (NF) and CAF. Results: CAF promoted the proliferation of A549 cells, while NF had no significant effect on them. After 72 hrs incubation, the absorbance value of A549+ CAF medium group was 0.814±0.006, significantly different from the 0.753±0.006 of the A549+ NF medium group (P<0.05). The Q-PCR assay indicated that mRNA expressions of Bcl-xL in the A549 group, A549+ NF medium group and A549+ CAF medium group were 1.00±0.11, 1.10±0.09 and 3.50±0.30, respectively, showing a significant difference between the A549+ NF medium group and A549+ CAF medium group (P<0.05). The Western blot showed that protein expressions of Bcl-xL in the A549 group, A549+ NF medium group and A549+ CAF medium group were 1.00±0.08, 1.10±0.12 and 3.10±0.25, respectively, with a significant difference between the A549+ NF medium group and A549+ CAF medium group (P<0.05). The ELISA results showed that the SDF-1 concentrations in the A549+ NF medium group and A549+ CAF medium group were 3.23±0.02 and 9.53±0.10, respectively, significantly different from each other (P<0.05). The MTT assay indicated that the absorbance values of OD of A549 group, A549+ AMD3100 group, A549+ NF medium group, A549+ NF medium+ AMD3100 group, A549+ CAF medium and A549+ CA Fmedium+ AMD3100 group were 0.43±0.03, 0.25±0.02, 0.48±0.03, 0.31±0.03, 0.72±0.06 and 0.45±0.03, respectively. The data of A549+ NF medium group was significantly different from that of A549+ CAF medium group (P<0.05). Conclusions Cancer-associated-fibroblasts enhance the drug resistance of A549 cells through SDF-1 secretion, upregulating the expression level of Bcl-xL through interaction with CXCR4. Our study not only illustrates that tumor microenvironment is able to enhance drug resistance of tumor, but also provides experimental evidence for the cancer-associated-fibroblasts as a potential therapeutic target for the treatment of lung cancer.

Non-conventional yeasts such as Yarrowia lipolytica, Pichia pastoris, Kluyveromyces marxianus, Rhodosporidium toruloides and Hansenula polymorpha have proven to be efficient cell factories in producing a variety of natural products due to their wide substrate utilization spectrum, strong tolerance to environmental stresses and other merits. With the development of synthetic biology and gene editing technology, metabolic engineering tools and strategies for non-conventional yeasts are expanding. This review introduces the physiological characteristics, tool development and current application of several representative non-conventional yeasts, and summarizes the metabolic engineering strategies commonly used in the improvement of natural products biosynthesis. We also discuss the strengths and weaknesses of non-conventional yeasts as natural products cell factories at current stage, and prospects future research and development trends.
Yeasts/genetics* ; Yarrowia/metabolism* ; Gene Editing ; Metabolic Engineering