Escherichia coli cytosine deaminase ( CD) gene was transfected into murine B16F10 melanoma cells by recombinant adenovirus AdCD in vitro . The tumor cells infected with AdCD were more sensitive to 5-fluorocytosine (5FC) than cells infected with a control adenovirus AdLacZ. The supernatant from B16F10 cells treated with AdCD/5FC was transferred to uninfected cells, and we found that only 6. 25 % of the supernatant could significantly inhibit the growth of wild type B16F10 cells. When AdCD was directly injected into established subcutaneous B16F10 tumors in mice followed by intraperitoneal injection of 5FC for 10 days, a significant reduction in tumor size and prolongation of survival period were observed. These studies not only explored the cytotoxic effects of AdCD/5FC on B16F10 melanoma cells in vitro and in vivo but also elucidated the mechanisms of its bvstander effect.

It was an important strategy in cancer gene therapy that transferring foreign gene to fibroblasts to express synthetic protein to exert antitumor effect. But most of studies regarded the foreign gene-modified fibroblasts as a by-stander, only a delivery source of synthetic protein, and neglected their irnmunological characters. In this paper, human primary dermal fibroblasts were modified by human IL-2 gene by retrovirus vector and induced by human IFN-?. The results showed that the MHC- I , MHC- II and CD40 expression on the surface of IFN-? induced fibroblasts were up-regulated significantly in comparison with those of non-induced cells. The IL-2, IL-1, IL-6 secretion were detected as a increased level in supematants of the IFN-?-induced, IL-2 gene modified fibroblasts. Because these molecules and cytokines play important roles in antigen presentation and effector activation, it can be inferred that the IFN-y induced, IL-2 gene modified fibroblasts could be used as antigen presenting cells in addition to delivery cells to activated the effector cells.

Objective To investigate the correlations between eosinophil counts in induced sputum and lung function (FENO) and evaluate these parameters in medication adjustment in patients with asthma.Methods Sixty-five outpatients with mild to moderate persistent asthma ( mild,32 ; moderate,33 ) from January to August 2008 were enrolled in the study.All were treated with combined medications comprising inhaled corticosteroids plus long-acting β2 agonists for 1 year.Lung function (FEV1％ and PEF％ ),eosinophil counts in induced sputum,FENO,and Asthma Control Test (ACT) scores were obtained at regular follow-up intervals.Twenty-one healthy volunteers served as controls,and lung function,eosinophil counts in induced sputum,and FENO were also obtained.Results Sixty-three subjects completed 1-year or longer follow-up.Lung function of 63 subjects recovered quickly in the early days and improved slowly during the following 6 months.FENO decreased from (61 ± 25 ) nmol/L at baseline to ( 32 ± 19 ) nmol/L by the third month (q =7.32,P＜0.05) and to (22 ± 12) nmol/L by the sixth month,which showed significant difference from normal controls [ ( 13 ± 8) nmol/L; q =6.63,P ＜ 0.05 ].Eosinophil counts in induced sputum of the asthma group at baseline were (0.093 ±0.023) × 109/L and decreased to (0.032 ±0.011)× 109/L by the third month,which was significantly different from baseline and normal controls [ (0.005 ±0.003) × 106/ml; q =5.49,P ＜0.05 and q =5.87,P ＜0.05,respectively].FENO showed a significantly positive correlation with eosinophil counts in induced sputum in the first 6 months (r1 =0.612,r2 =0.558,r3 =0.675; all P＜0.05) and a negative correlation with FEV1 (r1 =-0.537,r3 =-0.658,r6 ＝ -0.623,r9 =-0.537,r12 =-0.597 ; all P ＜0.05 ) at any time point of the study.The ACT score of 63 subjects at baseline was 14 ±3,and the scores after treatment for 1,3,6,9,and 12 months were 18 ±5,19 ±7,23 ±2,24 ± 1,and 24 ± 1,respectively; at the same time,significant difference was found ( F =5.72,P ＜ 0.05).Effectiveness was found according to the ACT score only 1 month after treatment.Conclusion The parameters of FENO and eosinophil counts in induced sputum were sensitive in the detection of airway inflammation and may be useful in evaluation of the efficacy of treatment and adjustment of medication regimens.

AIM: To investigate relationship between activity of matrix metalloproteinases - 2 ( MMP - 2, 72 kD) and invasion, metastasis of breast cancer. METHODS: Useing zymography and computer software assisted analysis, the activitive levels of MMP- 2 (72 kD) in tissues from breast cancer were measeured. RESULTS: Mean activitive levels of MMP- 272 kD (13.93 + 3.60) in breast cancer were lower than those in benign disease (21.43 + 8.31), P < 0.05. There was no difference (P > 0.05) in MMP - 2 62 kD + 72 kD of benign and malignant dis ease, but MMP - 262 kD ( 13.83 + 4.53) and MMP - 262 kD/62 kD + 72 kD (0.48) respectively were significantly higher in malignant disease (P < 0.01). It was also found that MMP- 262 kD/62 kD + 72 kD were apparently higher in invasive carcinomas (0.48) and lymph node metastases (0.61), P < 0.01, respectively. CONCLUSION: These results demonstrated that a clear relationship between MMP - 2 activity and the invasion and metastasis of breast carcinoma.

Nonaqueous capillary electrophoresis is used for the determination of the contents of diosgenin and ruscogenin in Radix Ophiopogonis. The operating buffer was composed of 20 mmol x L(-1) Na2B4O7-HCl (pH 7.61) in 70% methanol. The applied voltage was 25 kV and detection potential was at +0.70 V. With these conditions, the components were successfully separated. The content of diosgenin in Radix Ophiopogonis was 0.018 mg x g(-1) and ruscogenin was 0.008 mg x g(-1). The average recoveries of diosgenin and ruscogenin were 102% and 99.2%, respectively. A new method of the quality control of diosgenin and ruscogenin in Radix Ophiopogonis is provided.

AIM: To investigate relationship between activity of matrix metalloproteinases-2 ( MMP-2, 72 kD) and invasion, metastasis of breast cancer. METHODS: Useing zymography and computer software assisted analysis, the activitive levels of MMP-2 (72 kD) in tissues from breast cancer were measeured. RESULTS: Mean activitive levels of MMP-2 72 kD (13.93?3.60) in breast cancer were lower than those in benign disease (21.43?8.31), P0.05) in MMP-2 62 kD+72 kD of benign and malignant disease, but MMP-2 62 kD (13.83?4.53) and MMP-2 62 kD/62 kD+72 kD(0.48) respectively were significantly higher in malignant disease (P

AIM: To clone and express mouse canstatin (m canstatin)cDNA and provide a basis for the further research on its anti-angiogenic activity and potential application for cancer therapy. METHODS: Total RNA was extracted from mouse liver tissue by Trizol Reagent, and mouse canstatin cDNA was amplified by RT- PCR, then cloned into vector pMD18-T for sequencing. pET30a(+)-m canstatin recombinant plasmid was constructed and expressed in E.coli BL21 with induction of IPTG. RESULTS: Mouse canstatin cDNA is 684 bp coding 227 amino acids. The sequences of both cDNA and amino acid share high homology with human canstatin, with cDNA identity at 89% and amino acids identity at 96% to human canstatin. In the present study, pET30a(+)-m canstatin recombinant plasmid was expressed in E.coli BL21. CONCLUSION: Mouse canstatin cDNA has been cloned for the first time. Constructed pET30a(+)-m canstatin recombinant plasmid is highly expressed in E.coli BL21.

Adenoviruses harboring E. coli cytosine deaminase gene (AdCD) were used to transfect murine FBL-3 ery-throleukemia cells in vitro. FBL3 cells infected with AdCD were more sensitive to 5-fluorocytosine (5-FC) than cells infected with a control adenovirus AdLacZ. Further study indicated that this combination therapy (AdCD and 5-FC) killed tumor cells by inducing apoptosis of FBL-3 cells. The supematants from FBL-3 cells treated with AdCD/5-Fc were transferred on the culture system of uninfected (wild - type) FBL-3 cells, the result indicated that only 6.25% of the supernatant could induce significant cytotoxicity on wild type FBL3 cells. The results demonoustrated that bystander effect plays an important role in AdCD-mediated cytotoxicities. Direct injection of AdCD into established subcutaneous FBL3 tumor in mice followed by daily intraperitoneal injection of 5-FC for 10 days was found to inhibit tumor growth significant-

Antitumor effect of combined transfer of suicide gene and cytokine gene was evaluated in the present study. Adenoviruses expressing E. coli. cytosine deaminase (AdCD) and adenoviruses expressing murine interleukin 2 (AdTL2) were used for the treatment of tumor-bearing mice. The mice were inoculated s. c. with FBL-3 leukemia cells and 3 days later received intratumoral injection of AdCD in the presence or absence of AdIL2 followed by intraperitoneal 5-fluorocytosine (5FC) administration. The results demonstrated that tumor-bearing mice treated with AdCD/5FC in combination with AdTL2 showed more .potent inhibition of tumor growth and survived much longer as compared with mice treated with AdCD/5FC, AdEL2, AdlacZ/5FC or PBS. It was illustrated that the tumor mass showed obvious necrosis and inflammatory cell infiltration, and more CD4+ and CD8+ T cells infiltrated into the tumor after combined therapy. The splenic NK and CTL activities increased significantly in mice after combined transfer of CD gene and EH gene. Our results demonstrated that combined transfer of suicide gene and IL-2 gene could inhibit the growth of established tumor in mice significantly and induce antitumor immunity of the host efficiently.

Aim To identify whether the petroleum e-ther fraction of Gardenia jasminoides Ellis ( GJ-PE ) could effetive exhibit a rapid antidepressant effect and also to investigate the biological mechanism. Methods Tail suspension test ( TST ) , forced swimming test ( FST ) and novelty suppressed-feeding ( NSF ) were used to screen the rapid antidepressant potential of ef-fective fractions of GJ-PE in KM mice at 24 h post a single administration. Tail suspension test ( TST) was also used at 30 min and forced swimming test ( FST ) was used at 2 h to test the initial onset time of effective fractions of GJ-PE in KM mice. Western blot was per-formed to examine the expression of BDNF and p-eEF2 in hippocampus of KM mice at 2 h and 24 h. Results An acute administration of GJ-PE1 decreased the im-mobility time of KM mice in FST at 2 h and 24 h and decreased the latency time in NSF at 24 h. GJ-PE3 de-creased the latency time in NSF at 24 h. GJ-PE4 in-creased the unit food consumption in NSF at 24 h. At 2 h post a single GJ-PE1 treatment, the expression of BDNF was significantly up-regulated while the expres-sion of p-eEF2 was significantly down-regulated. At 24 h post a single GJ-PE1 treatment, the expression of BDNF was significantly down-regulated while p-eEF2 expression was significantly up-regulated. Conclusion GJ-PE1 has the most significant rapid antidepressant potential among the four fractions of GJ-PE. The effec-tive time of GJ-PE1 is 2 h after drug treatment. The mechanism of the rapid antidepressant effect of GJ-PE1 at 2 h is related to the up-regulation of BDNF and down-regulation of p-eEF2 . GJ-PE3 and GJ-PE4 also have some features of rapid antidepressants. GJ-PE2 doesn′t have the rapid antidepressant potential.