IL-3 has effects on a wide variety of cell types, including immature cells of the immune cells, and mature immune cells such as granulocytes. In the purpose of studying the immunoregulatory function of IL-3 and its potential role in cancer therapy, we established a IL-3-secreting tumor model using gene transfection to deliver locally IL-3 to tumor site. First, we constructed IL-3 expression vector BMGNeo-mIL-3, then transfected it into B16 murine melanoma cells. By G418 resistant screening and limiting dilution, we obtained a transfectant that expressed high levels of IL-3 (806U / ml) . The IL-3 expression of the transfectant was confirmed by Northern blot analyses. Although the wild-type B16 cells and the B16-Neo cells transfected with BMGNeo did not express IL-3, the IL-3 expression in B16 cells had no obvious effect on the in vitro proliferation of the transfectant. These results showed we had successfully established a IL-3-secreting tumor cell clone which would enable us to further study its in vivo tumorigenicity and immune function.

In the present study, we prepared cationic liposomes encapsulated IL-2 DNA or IL-6 DNA with reverse-phase evaporation to investigate their efficiency in mediating gene transfer. After transfection with IL-2 DNA liposome or IL-6 DNA liposome, the supernatants of NIH3T3 cell contained high levels of IL-2 or IL-6. The peritoneal cells from the mice injected i.p. of IL-2 DNA or IL-6 DNA liposome 10 days later could also be detected to secrete IL-2 or IL-6. The B16F10 melanoma cells from tumor-bearing mice which were injected intratumorally of IL-2 DNA or IL-6 DNA liposome exhibited resistant to G418 and secreted high levels of IL-2 or IL-6. The results show that the liposome can mediate cytokine gene transfer efficiently in vitro and in vivo.

The Ad-IL-2 and/or Ad-IL-3 were i. p. injected into experimental leukemic mice after high dose chemotherapy. When Ad - LacZ was i. p. injected into leukemic mice, there were leukemic cells in bone marrow, vessal, liver and spleen. After i. p. injection of Ad-IL-2 and/or Ad-IL-3, the leukemia - bearing mice showed significant inhibition of tumor growth. As i.p. injection of Ad-IL-2, the splenic NK, CTL activity increased; as i. p. injection of Ad - IL - 3, the number and cytotoxicity of peritoneal macrophages were significantly higher than that of the control groups. After i. p. injection of Ad - IL - 2 and Ad - IL - 3 at the same time, the anti - leukemia activity was the best. Though there were less leukemic cells in the bone marrow, there were no leukemic cells in the liver and spleen. Our results suggested that i. p. injection of Ad - IL - 2 and Ad - IL - 3 could significantly enhance the anti - leukemia activity of chemotherapy.

The human Interleukin-2 cDNA containing full-length of encoding region was cloned by RT-PCR from PBMNC and confirmed by DNA sequencing. The hIL-2 recombinant retroviral expressing vector (pLXSN-hIL2) was constructed by inserting hIL-2 cDNA into the BamHI cloning site of pLXSN retroviral vector. After packaged with CRIP packaging cell line, the hIL-2 retrovirions were produced at the titer of 7.6?105CFU/ml. Then a fibroblast cell clone(NIH3T3-hIL2) secreting 118.2U/ml hIL-2 was obtained by infecting the NIH3T3 fibroblast cells with hIL-2 retrovirions. The integration of hIL-2 provirus into the genome of NIH3T3-ML2 cells was confirmed by PCR analysis for NeoR gene. Our data showed that the hIL-2 retroviral vector was successfully constructed, which enables us to further the investigation of the hIL-2 gene therapy in clinical trial.

To evaluate whether in vitro and in vivo transfer of E. Coli cytosine deaminase gene will confer sensitivity of a solid tumor to prodrug 5-fluorocytosine(5FC), we used an adenovirus vector(AdexCMV. CD) carrying the cytosine deaminase gene driven by the CMV promoter, infected SMMC-7721 or HepG2 cells hepatocellular carcinoma cells in vitro, and found AdexCMV. CD vector could effectively suppressed the growth of SMMC-7721 and HepG2 cells. When the two cells were infected with AdexAFP. CD vector in which the CD gene was driven by the AFP gene 5'-flanking region, only HepG2 cells were conferred sensitivity to 5FC. (Infection with AdexCMV.CD, when as few as 20% of cells transfected the CD gene, SMMC-7721 cells were associated with a bystander effect when combined with 5FC in cell mixing studies.) Consistent with these in vitro observations, AdexCMV. CD was directly injected into established subcutaneous SMMC-7721 tumors in nude mice receiving 5FC,there was a 60% reduction in tumor size at day 8, 70% reduction at day 24. Our results suggested that adenovirus-mediated tumor-specific gene transfer of CD gene and concomitant administration of 5 FC may have potential as a strategy for local control of tumor growth.

The human CD80 full-length encoding cDNA was cloned by RT-PCR, and inserted into El-substituted aden-ovirus vector pAxlcw. Subsequently, the hCD80 recombinant adenovirus vector was cotransfected into 293 cells together with EcoT22I-digested Ad5-TPC, and the replication-deficient hCD80 recombinant adenovirus was generated efficiently by homologous recombination, with the tilers of 4?10~9pfu/ml. Infected with prepared hCD80 recombinant adenovirus in vitro, Hela cells expressed high levels of hCD80 48 hours after infection. These suggest that COS/TPC is an efficient method to prepare recombinant adenoviruses and the prepared hCD80 adenovirus could be used in cancer gene therapy.

Objective To investigate clinicopathologic characteristics of malignant tumors initiated with cervical lymph nodes enlargement.Methods Clinical and pathological data of 887 cases with cervical malignant tumors kept in the college during January 1998 to December 2003 were analyzed for their histological patterns and primary site of metastatic carcinoma.Results Cervical cancer with lympb nodes metastasis was mostly seen at age of 51~60 years (182/837,21.74%), and much more common in men than in women. Major metastatic carcinomas in cervical lymph nodes originated from the nasopharynex, lung, thyroid, stomach and esophagus (591/671,88.08%), with 60.66%(407/671)of squamous cell carcinoma mostly from the nasopharynex, lung, or esophagus (380/407,93.37%). There was significant difference in the site, size, number, quality, appearance of lymph nodes, course of disease, and clinical manifestations between lymphoma and metastatic cancer.Conclusions Cervical metastatic carcinomas in Guangdong mainly originated from the nasopharynx. Some specific immunohistochemical markers, CT, MRI and ultrasonic technology played a very important role in diagnosis for lymphoma and determination of primary site of metastatic cancer.

Objective To investigate the correlation between levels of glycohemoglobin and cognitive function in elderly patients with type 2 diabetes. Methods Cognition function of 40 elderly patients with type 2 diabetes was assessed by Wechsler Memory Scale (WMS), Trail Marking Test and Wisconsin Card Sorting Test (WCST) and was compared with 40 healthy controls matched for age, gender, education level and profession. Meanwhile, the correlation between cognition function and levels of glycohemoglobin was analyzed by regression analysis.Results The scores of Wechsler Memory Scale in the patients were lower than that in normal controls (P＜0.01，P＜0.05). The patients needed much more time to complete the Trail Marking Test (P＜0.001). The scores of Wisconsin Card Sorting Test were significantly different between two groups (P＜0.01，P＜0.05). The regression analysis showed that there was an obvious correlation between serum level of glycohemoglobin and cognition functions in patients with type 2 diabetes(r=-0.38, -0.43, -0.48, -0.46, 0.41, P＜0.05). Conclusion Cognition function decreases and significantly correlates with the level of glycohemoglobin in the elderly patients with type 2 diabetes.

A novel gene, which was a homologue of Arabidopsis COI1 was isolated from rice (Oryza sativa L.) by RT-PCR and designated as OsCOI1. It encoded a protein of 595 amino acids. The similar F-box motif and 16 leucine-rich repeats were found in the deduced protein OsCOI1. OsCOI1 and COI1 showed high homology (74%) at amino acid level. Semi-quantitative RT-PCR and Northern blot analysis demonstrated that the expression of OsCOI1 in rice varied obviously after treatment with MeJA and ABA but was not affected by SA and ET, suggesting that the specific function of OsCOI1 in JA signal pathway and related ABA pathway.

Objective To investigate the clinical utility of anti-dsDNA antibody in the diagnosis of systemic lupus erythematosus (SLE).Methods Total and high affinity anti-dsDNA antibodies of 431 serum samples were measured by total and high affinity anti-dsDNA antibody EIA kits.Modified SLE disease activity index (M-SLEDAI) was scored for each SLE patients at the time of serum collection.The agreements between total and high affinity anti-dsDNA EIA tests and the correlation of anti-dsDNA antibody levels with disease activity and clinical manifestations were analyzed.Statistical analysis was performed using x2 test,Spearman correlation,Mann Whitney U test & Fisher exact,Student t test.Results ① The overall agreement between total and high affinity anti-dsDNA antibodies was 93.3％,and the total agreement of SLE patients was 89.6％.Twenty-two out of 23 (95.7％) SLE patients with positive total anti-dsDNA antibody but high affinity antidsDNA antibody were inactive with the M-SLEDAI score less than 4.Four patients with other autoimmune diseases had positive total anti-dsDNA antibody but negative on high affinity anti-dsDNA antibodies.② The total and high affinity anti-dsDNA antibody levels were significantly positively correlated with disease activity (M-SLEDAI) and negatively correlated with serum C3 and C4 levels (P＜0.01).③ The ratio of high affinity to total anti-dsDNA antibody was significantly higher in patients with M-SLEDAI ≥4 or with active kidney damage (P＜0.01).Conclusion Both total and high affinity anti-dsDNA antibodies are significantly correlated with SLE disease activity and kidney damage.However,the high affinity anti-dsDNA antibody may be more specific for active SLE and helpful in the differential diagnosis with other autoimmune diseases.High affinity anti-dsDNA antibody is more valuable in monitoring SLE disease activity status,especially on kidney damage,compared to total anti-dsDNA antibodies.