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OBJECTIVE To investigate the effects of soybean isoflavones （SI） on the reproductive development of young mice. METHODS C57BL/6 young mice were randomly divided into control group， SI low-dose and high-dose groups （10， 100 mg/kg）， with 10 mice in each group （half male and half female）. The young mice in each group were given corresponding liquid intragastrically， once a day， for 2 consecutive weeks. After the last administration， the percentage of body weight increase was calculated； serum estradiol and testosterone levels， malondialdehyde （MDA） content， total antioxidant capacity （T-AOC）， superoxide dismutase （SOD） and glutathione peroxidase （GSH-Px） activities in the reproductive organs of the young mice were determined. The histopathological changes in the reproductive organs were observed. The cell apoptosis of reproductive organs was detected. RESULTS Compared with the control group， the percentage of body weight increase in female mice was increased significantly in the SI high-dose group， while that of male mice was decreased significantly （P＜0.05 or P＜0.01）. Cystic follicles could be seen in the ovarian tissue in SI groups， a loose arrangement of spermatocytes could be seen in the testicular tissue， and partial epithelial cell shedding could be seen in epididymal tissue. The serum level of testosterone in female young mice and the serum levels of testosterone and estradiol in male young mice in SI groups， GSH-Px activity in the ovarian tissue of female young mice in the SI low-dose group， T-AOC activities in the ovarian tissue of female young mice in SI groups as well as the apoptotic rates of cells in testicular and epididymal tissue of male young mice in SI groups were increased significantly （P＜0.05 or P＜ 0.01）； the serum level of estradiol in female young mice in SI groups， SOD activity in the ovarian tissue of female young mice in the SI high-dose group， and MDA contents in the ovarian tissue of female young mice in SI groups as well as the apoptotic rates of cells in ovarian tissue of female mice in SI groups were decreased significantly （P＜0.05 or P＜0.01）. CONCLUSIONS SI can enhance the antioxidant stress capacity of ovarian tissue in female young mice and reduce their oxidative stress damage， but it has certain toxicity to reproductive organs in male mice.
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Objective To optimize conditions for the preparation of amidoximated polyacrylonitrile (PAO) membrane for U (VI) adsorption that can be easily recycled and reused. Methods Using polyacrylonitrile powder as a raw material, the PAO membrane was prepared using the nonsolvent-induced phase separation method under different conditions. The effects of the polyacrylonitrile concentration and temperature and duration of amidoximation in the preparation of casting solution as well as alkali treatment before adsorption on the U (VI) adsorption performance of PAO membrane were investigated. Results Alkali treatment significantly improved the U (VI) adsorption performance of PAO membrane, and the temperature and duration of alkali treatment affected the U (VI) adsorption capacity. The optimal alkali treatment conditions were 0.1 mol/L NaOH, 80 min, and 60℃. The polyacrylonitrile concentration and temperature and duration of amidoximation in the preparation of casting solution influenced the adsorption performance of PAO membrane. The optimal preparation conditions were 10wt% polyacrylonitrile and amidoximation at 65℃ for 18 h. Under the optimal preparation and alkali treatment conditions, the U (VI) adsorption capacity of PAO membrane reached 255 mg/g. Conclusion The PAO membrane has the advantages of simple preparation, high adsorption capacity, and easy recycling and reuse, which can be used as an ideal material for adsorbing U (VI) in wastewater.
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Objective To observe the inhibitory effect on lymph node metastasis of pancreatic cancer and lymphangiogenesis in mice by injection of KAll gene within xenograft tumor.Methods Pancreatic cancer cell line MiaPaCa-2 wag used to construct the nude mice models bearing tumors,then the mice were divided into normal saline group,Ad group and Ad-KAI1 group.Since the successful model construction,normal saline,Ad,Ad-KAI1 was injected every week for 3 times,respectively in the three groups,then the tumor size was documented.50 d after model construction,the tumor and enlarged lymph nodes were collected and subjected to pathological exam,and the expression of LYVE-1 and the MLVD in xenograft tumor was detected by immunohistochemistry.Results Two weeks after MiaPaCa-2 implantation,the model was 100％ successfully constructed.The growth curve of subcutaneous tumor among 3 groups was not statistically significant (P＞0.05) ; the weights of subcutaneous tumor in the 3 groups were (2514.4 ±351.3),(2466.1 ± 295.5),(2294.4±255.4) mg after 50 d,and the difference among the 3 groups was not statistically significant (P ＞0.05).Enlarged lymph nodes metastasis was observed in 8 mice (80％) in normal saline group,and 20 lymph nodes were collected,with 2.0 lymph nodes per mice; and enlarged lymph nodes metastasis was observed in 7 mice (70％) in Ad group,and 15 lymph nodes were collected,with 1.5 lymph nodes per mice; while enlarged lymph nodes metastasis was observed in 4 mice (40％) in Ad-KAI1 group,and 6 lymph nodes were collected,with 0.6 lymph nodes per mice.All the lymph nodes were confirmed to be metastasis of the primary tumor after pathologic exam.The difference of lymph nodes metastasis,number of lymph nodes metastasis per mice among the 3 groups was statistically significant (F =3.14,3.35,P ＜ 0.05).The MLVD of subcutaneous tumor among the 3 groups was (18.70 ± 5.60),(19.40 ± 4.58),(9.80 ±4.10)/400 times magnification,the MLVD of Ad-KAI1 group was significantly lower than those in normal saline group and Ad group (F10.76,11.36,P ＜ 0.05),but the difference between normal saline group and Ad group was not statistically significant.Conclusions KAI1 can inhibit the lymph node metastasis of pancreatic cancer,and the mechanism may be related with decreased lymphangiogenesis and reduced lymphatic vessel density.
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Objective To investigate the ability of induction of specific cytotoxic T lymphocytes (CTL) stimulated by dendritic cells (DCs) co-transfected with MUC1 and survivin mRNA of human pancreatic cancer,and to provide the experimental basis for the treatment of human pancreatic cancer with multi-epitope DC vaccine.Methods DCs were isolated and cultured from peripheral blood mononuclear cells (PBMCs) of 6 patients with pancreatic cancer.Human pancreatic cancer cell line MiaPaCa-2 was routinely cultured,after being transcripted and amplified by RT-PCR,MUC1 and survivin mRNA were co-transfected or individually transfected into DCs by electroporation,and they were named as DC-MUC1,DC-survivin,DC-MUC1 + survivin.The expression of MUC1 and survivin mRNA in DCs were detected by real-time PCR.The survival rate of transfected DCs were determined by MTT method.The lymphocyte proliferation ability was evaluated by mixed cell culture method.The Th1 cytokine releasing of antigen-specific CTLs were measured by ELISA assay.Results Mature DCs were obtained,the positive expression rates of surface markers CD40,HLA-DR,CD83 and CD86 were 34.31％,50.21％,89.17％ and 73.62％,respectively.The expression amount of MUC1 mRNA of DC-MUC1 was 36.24 ± 5.17,and the expression amount of survivin mRNA of DC-survivin was 34.53 ± 4.02,while the expression amounts of MUC1,survivin mRNA of DC-MUC1 + surviving were 31.79 ±4.26 and 14.67 ± 2.96,which were significantly lower than that in individual transfection group (P ＜ 0.05).The survival rate of DC-MUC1 + surviving was decreased in a time dependent manner,which was significantly lower than that in individual transfection group (about 50.21％ vs 80％ at 24 h,P ＜0.05).When DC/T cells ratio was 1∶ 10,1∶ 20,the autologous T cell proliferation index of MUC1 and survivin mRNA in co-transfection DC group was significantly higher than that in individual transfection group (P ＜ 0.05) ;when DC/T cells ratio was 1∶ 40,1∶ 80,the difference of proliferation index was not statistically significant.When DC/T cells ratio was 1∶ 10,after 14 d culture,the expressions of IL-2 in DC-MUC1,DC-survivin,DC-MUC1 + surviving were (892.73 ± 32.9),(713.62 ± 56.37),(1884.37 ± 95.21) pg/ml,and the expressions of granzyme B were (501.62 ± 12.30),(203.84 ± 12.55),(1193.15 ± 86.04) pg/ml ; and the expressions of IFN-γ were (981.50 ± 47.82),(696.05 ± 41.66),(2237.94 ± 189.55) pg/mL.The corresponding values in DC-MUC1 + surviving group were significantly higher than those in individual transfection group (P ＜ 0.05) ; while the difference of IL-10 was not statistically significant.Conclusions DCs co-transfected with MUC1 and survivin mRNA have a stronger ability to stimulate specific CTL in vitro than individual antigen loaded DCs.
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Objective To investigate the effects of KAll gene transfection on proliferation,migration,invasion and VEGF expression of pancreatic cancer MiaPaCa-2 cells under hypoxic condition,and explore possible mechanism.Methods The pcMV-KAI1 vector which contained the full length of KAI1 cDNA was transfected into pancreatic cancer cells MiaPaCa-2,and KAI1,VEGF C,VEGF A protein expressions were determined by Western blot.The proliferation of pancreatic cancer cells was evaluated by MTT method.Wound-healing assay and cell invasion assay were used to detect the migration and invasion of pancreatic cancer cells.The expression of VEGF C,VEGF A in supernatant of culture was measured by ELISA method.Results The expression of KAI1 protein in MiaPaCa-2 K transfected with KAI1 was significantly higher than that in nontransfected cells [(0.549 ± 0.021) vs 0,P＜0.05].The proliferation under hypoxic condition was not significantly different,but the migration distance was significantly shorter and the number of transmembrane cell was significantly decreased [(14.0 ± 5.8) vs (43.0 ± 14.4),P ＜ 0.05].The expression of VEGF-C in cell was significantly decreased [(0.218 ± 0.043) vs (0.745 ± 0.069),P ＜0.05],but the expression of VEGF-A was not significantly different; the expression of VEGF-C in cell culture supernatants was significantly decreased [(1236 ± 247) vs (2045 ± 221) pg/ml,P ＜ 0.01].Conclusions The migration,invasion ability of pancreatic cancer MiaPaCa-2 cells with KAll transfection under hypoxic condition is decreased,and the possible mechanism of inhibiting lymphatic metastasis is down-regulation of VEGF-C expression.