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BACKGROUND: As a combination of reverse transcription (RT) and polymerase chain reaction (PCR), RT-PCR has been used to detect gene expression levels in cells and tissues, RNA virus contents in cells and specific gene cloned cDNA sequences.OBJECTIVE: To detect the inhibitory effcet of Stealth siRNAs on the expression of transforming growth factor β1 (TGF-β1).METHODS: There were blank control, empty vector transfection, stealth-48, stealth-166, and stealth-594 groups. Three stealth siRNAs aimed at different sequences in TGF-β1 mRNA were made, and were then transfected into BALB/c mouse lung fibroblasts in vitro. The expressions of TGF-β1 and connective tissue growth factor were detected by RT-PCR.RESULTS AND CONCLUSION: In different time periods, the TGF-β1 expression was differentially depressed by three stealth siRNAs, especially stealth-166. The inhibitory effects varied with time, which could be detective at 48 hours,reached the peak at 72 hours and then began to attenuate at 96 hours. Our findings show that the inhibitory effect of stealth siRNAs on the TGF-β1 expression in mouse lung fibroblasts can be detected by RT-PCR.
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Objective:To determine the inhibitory effect of Rupixiao ( RPX) granule on mammary gland hyperplasia ( HMG) in rabbits and explore the possible mechanism to provide reference for clinical medication. Methods:Rabbit model of mammary hyperpla-sia was established by estradiol benzoate and progesterone. Xiaoyao pills and tamoxifen were used as the positive control, and the RPX granule group was respectively at low, medium and high dose (0. 525,1. 05,2. 1 g·kg-1). Each group was with intragastric adminis-tration for 30 days. The levels of E2 , PROG, FSH, LH and PRL in serum and the expression of VEGF in the homogenate tissue were determined by an Elisa method. The breast tissue of rabbits in each group was withdrawn to observe the structure changes after Hema-toxylin-eosin staining ( HE) . Results:Ovarian preservation-benzoic acid, estradiol combined with progesterone could be used to estab-lish mammary gland hyperplasia in rabbits. RPX granule could significantly decrease the serum levels of E2 and PRL (P<0. 01)and increase that of PROG (P<0. 01 or P<0. 001). The results of pathological section showed that the breast structure in RPX granule high dose group could be restored to the level of the blank control group, which showed its therapeutic effect on HMG was better than that of tamoxifen and Xiaoyao pills. The expression of VEGF in each treatment group significantly decreased compared with that in the model group by the Elisa reagent (P<0. 001). Conclusion:RPX granule has good efficacy in HMG rabbits. The inhibitory effects of RPX granule on HMG in rabbits maybe related to the inhibition of VEGF in rabbit breast tissues, and the inhibition of RPX granule was similar to that of tamoxifen and Xiaoyao pills.
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Objective To evaluate the effects of the recombinant human interleukin-1 receptor antagonist(rhIL-1ra)on a tol?uene-2,4-diisocyanate(TDI)-induced guinea pig allergic rhinitis (AR)model. Methods An AR model was established via sensiti?zation and challenge of two-step procedure using TDI in guinea pigs. Normal animals were treated only with the olive oil(TDI vehicle). Sixty adult guinea pigs were randomly divided into six groups(n=10):normal group,model group(rhIL-1ra vehicle),positive con?trol group(budesonide,25.6μg/kg),rhIL-1ra treated groups(rhIL-1ra 50,100 and 200μg/kg,respectively). From day 8 after sensi?tization,animals of all the groups were treated respectively with the agents or vehicle once a day for 14 days. During the observation pe?riod,the index of clinic score was recorded for every animal. At day 14 of the dosing,guinea pigs were sacrificed 30 min after the last TDI challenge and observation. Blood samples were taken from the abdominal aorta to prepare the serum for detection of histamine , and the nasal mucosase were dissected for histamine detection and histopathological observation. Results Compared with the guinea pigs in normal group,those in the model group exerted the typical symptoms of AR. It was shown that rhIL-1ra could improve nasal symptoms and cause a significant decrease in the instances of nasal sneezing as well. In addition,rhIL-1ra significantly reduced the concentrations of histamine in the nasal mucosa and IgE in the blood compared with those in the model group(P<0.05). Moreover, the pathological results showed that less edema,vasodilation and inflammatory cell infiltration were found in the nasal mucosa after rhIL-1ra application. Budesonide also showed the above effects with no significant difference compared with rhIL-1ra. Conclusion A guinea pig allergic rhinitis model is successfully induced by TDI. The results indicated that rhIL-1ra(50-200μg/kg)is effective in im?proving allergic rhinitis. Our findings indicated that rhIL-1ra might serve as a potential new drug for allergic rhinitis therapy.
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Objective To improve the filtering technique and the addition methods of distillate for Tongtian oral liquid with a new type of filtering equipment. Methods The filter paper in the current filtering process was replaced with a combination of terylene filter cloth and pulvis talci. Distillate was added into the runny paste of preparation directly without using the solubilizer of Tween-80. Then the admixture was agitated sufficiently. After that, 10% NaOH was used to adjust the pH to 4.5-6.5, and the solution was added to 1200 L by water. At last, a clear preparation was obtained by using the above filtering equipment. Results Compared with the product made by the current method,the improved preparation was qualified and tasty. Conclusion The improved method is feasible, simple and economical. The product is tasty.
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Objective To reveal the alterations of metabolic profiles by detecting changes in hippocampus 1H NMR spectra in acute brain injury (ABI) rats. Methods The model of ABI was achieved by trauma, and 2 hours after modeling, the hippocampus samples were collected for NMR analysis. Results Compared to age-matched control rats, the ABI rats exhibited markedly different metabolic profiles, which showed significant changes in glutamate(P<0.01), taurine(P<0.05), myo-inositol(P<0.05), and glutamine(P<0.05). Conclusion The metabonomics results suggest that amino acid metabolism is disordered when ABI occurs. This study provides clues for further understanding to cure ABI.