The study of the relationship between trace elements and drug resistance of tumor has been going on for a long time.It is clear that some trace elements are related to the tumor occurrence and its progress, such as selenium,zinc,copper.So they can be used as an sub index to diagnosis and to evaluate prognosis. Further study in recent years has shown that trace elements are related to the tumor drug risistance. Improveing nourishment of tumor cells may change its drug resistance and influence the curative effect of tumor. The significance of this study is to search for a simple and approach in reducing drug resistance in order to improve the chemotherapeutic effect and the patient′s prognosis.

OBJECTIVE:To screen the optimal technology of dog testes and penis processed with talcum powder,and to pro-vide reference for the formulation of quality control standard. METHODS:Taking the comprehensive score of crushing rate and the content of alcohol soluble extract as evaluation indexes,L9(34) orthogonal test was used to optimize processing temperature,pro-cessing time and the amount of talcum powder;validation test was conducted. According to related method in Chinese Pharmaco-poeia(2015 edition),the contents of moisture,ash,nitrogen and alcohol soluble extract were determined,and automatic amino ac-id analyzer was used to determine the content of amino acids in samples. RESULTS:The optimal processing technology were that 100 kg dog testes and penis should add into 40 kg talcum powder;the processing temperature was 350-380 ℃;processing lasted for 4 min. Results of verification test showed that the average crushing rate and the content of alcohol soluble extract were 80.2%(RSD=0.95%,n=3)and 15.7%(RSD=2.30%,n=3). In processed dog testes and penis,the contents of moisture,ash,nitro-gen and alcohol soluble extract were 5.7%,18.7%,8.3%,15.7%,respectively. Besides,there still were 16 kinds of amino ac-ids,and their total content was 42.44%. CONCLUSIONS:The optimized talcum powder processing technology of dog testes and penis is stable and practical. The content of moisture,ash,nitrogen,alcohol soluble extract and amino acid can be used as impor-tant quality standard control indexes for processed dog testes and penis.

ObjectiveTo find out the inhibitory effects of CD4 - CD8 - DNT cells on growth of which depresses the pancreatic cancer in vitro and in vivo.Methods The inhibitory effects of DNT cells on the growth of Panc- 1 were studied in vitro by MTT method.Eighteen BALB/c mice were divided into 3 groups randomly.Human pancreatic cancer xenografts were established in 2 groups randomly.The last group was injected the cell suspension which comprises DNT and Panc- 1 cells ( Panc- 1∶ DNT =1∶ 5 ).When the diameter of tumor was about 5 mm,the first 2 group mice were further divided into 2 groups randomly.One was control,treated with distilled water.The other was treated with celebrex (4 mg/d).The size of the tumors was calculated every 2 weeks and tumor growth curve was depicted.At the end of the treatments,the mice were sacrific and the tumors were harvested.The tumor inhibition rate was calculated.Results( 1 ) MTT study showed that DNT cells produced a dose- dependent inhibition of Panc- 1 proliferation in vitro.(2) The growth of transplanted pancreatic cancer was down-regulated by treatment of DNT cells.ConclusionDNT cells can inhibit the growth of pancreatic cancer in vitro and in vivo.

Objective To explore the mechanism of anti-tumor effects of transferring tumor-specif-ic lymphocytes obtained from pre-immunized BALB/c mice with inactive rolL-21 tumor vaccine (mIL-21-Sp2/0)to syngeneic mice, associated with mIL-21 tumor vaccine immunization, in the condition of cyclo-phosphamide (Cy)-induced lymphopenia. Methods Activated lymphocytes of spleen and lymph nodes ob-tained from pre-immunlzed syngeneic mice with irradiated mIL-21-Sp2/0 cells were infused into BALB/cmice treated with Cy 2 days before, subsequently vaccinated with mlL-21 tumor vaccine, after 7 days, chal-lenged with Sp2/0 tumor cells, observed the growth of tumor of mice. T lymphocyte subsets differentiation was measured by flow cytometry (FCM) analysis. The proliferation and cytotoxie activities of activated lym-phocytes were analyzed by FCM, respectively, staining with CFSE and 7-AAD. The number of IFN-γ-secre-ting cells was evaluated by ELISPOT. Results The lymphopenic mice were transferred with activated lym-phocytes and inoculated with raiL-21 tumor vaccine might provide superior anti-tumor immunoprotection, re-tard tumor growth of the mice. The proliferating capabilities and killing rate of transferred tumor Ag-specific lymphocytes enhanced obviously, the number of IFN-γ-secreting cells was significantly higher compared with the control groups. Conclusion Under Cy-induced lymphopenia condition, tumor Ag-specific lymphocytes sensitized by raiL-21 tumor vaccine were transferred to mice and immunized with mlL-21 tumor vaccine at the same time, benefit the proliferation of transferred effective cells and immune cells itself, assist to form and sustain special anti-tumor effects.

Glycyrrhiza uralensis Fisch. ex DC is widely used in traditional Chinese medicine (TCM). Among its various active components, glycyrrhizic acid is believed to be the marker component. Squalene synthase (SQS) and beta-amyrin synthase (beta-AS) are key enzymes in the biosynthetic pathway of glycyrrhizic acid in G uralensis. To reveal the effects of co-expression of SQS1 and beta-AS genes on this pathway, 7 yeast expression vectors harboring different SQS1 variants and beta-AS were constructed and expressed in Saccharomyces cerevisiae as fusion proteins. TLC and GC-MS results showed that co-expression of SQS1 and beta-AS enhanced the accumulation of beta-amyrin. The effects of SQS12 were more obvious than the other two SQS1 variants. This study is significant for further investigations concerned with exploring the biosynthesis of glycyrrhizic acid in vitro and strengthening the efficacy of G. uralensis by means of increasing the content of glycyrrhizic acid.

The rate-limiting enzyme in the mevalonic acid (MVA) pathway which can lead to triterpenoid saponin glycyrrhizic acid (GA) is 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR). In order to reveal the effect of copy number variation in the HMGR gene on the MVA pathway, the HMGR gene from Glycyrrhiza uralensis Fisch. (GuHMGR) was cloned and over-expressed in Pichia pastoris GS115. Six recombinant P. pastoris strains containing different copy numbers of the GuHMGR gene were obtained and the content of ergosterol was analyzed by HPLC. The results showed that all the recombinant P. pastoris strains contained more ergosterol than the negative control and the strains with 8 and 44 copies contained significantly more ergosterol than the other strains. However, as the copy number increased, the content of ergosterol showed an increasing-decreasing-increasing pattern. This study provides a rationale for increasing the content of GA through over-expressing the GuHMGR gene in cultivars of G. uralensis.