目前,国内尚无将体外转译体系应用于IL 2研究的报道。本文以长春1号小麦种制备麦胚无细胞体系,经与兔网织红细胞、腹水癌细胞两体系比较筛选,结果证明麦胚无细胞转译体系适于实验室应用。通过对ML A 144mRNA作转译活性鉴定,证明该系统...目前,国内尚无将体外转译体系应用于IL 2研究的报道。本文以长春1号小麦种制备麦胚无细胞体系,经与兔网织红细胞、腹水癌细胞两体系比较筛选,结果证明麦胚无细胞转译体系适于实验室应用。通过对ML A 144mRNA作转译活性鉴定,证明该系统可以合成具有生物活性的、完整的IL2分子。展开更多
In this study, the IL-2 mRNA levels of T lymphocytes in normal mice stimulated by nine Chinese herbal medicinal ingredients （CHMIs） were measured using semiquantification reverse transcription polymerase chain react...In this study, the IL-2 mRNA levels of T lymphocytes in normal mice stimulated by nine Chinese herbal medicinal ingredients （CHMIs） were measured using semiquantification reverse transcription polymerase chain reaction. The results showed that astragalus polysaccharide （APS）, epimedium polysaccharide （EPS）, Chinese angelica polysaccharide （CAPS）, propolis flavone （PF）, and astrogalosides （AS） promoted IL-2 mRNA levels in T lymphocytes in vitro and in vivo to differing degrees, and the level of IL-2 mRNA induced by propolis polysaccharide （PPS） in vitro was higher than that induced by the control, which differed from that of PPS in vivo.展开更多
The purpose of the study is to establish a fluorescence quantitative reverse transcription polymerase chain response （FQ-RT-PCR） method for the quantitative determination of IL-2 mRNA and IL-4 mRNA in Th cells, with...The purpose of the study is to establish a fluorescence quantitative reverse transcription polymerase chain response （FQ-RT-PCR） method for the quantitative determination of IL-2 mRNA and IL-4 mRNA in Th cells, with which the Th cells status of the patients with gynaecological tumors and chronic renal failure （CRF） can be analyzed. IL-2 eDNA and IL-4 eDNA were prepared, and the plasmid pMD18 carrying IL-2 eDNA or IL-4 eDNA fragment was constructed and cloned as the template for quantitative determination. The primers and probes labelled with 6-carbexy-fluorescein （FAM） and 6-carboxy- tetramethylrhodamine （TAMRA） were prepared, and the experimental conditions were optimized to set up the FQ-RT-PCR method for quantitative determination of IL-2 mRNA and IL-4 mRNA. Th cells enriched from peripheral blood mononuclear cells （PBMCs） of 20 healthy volunteers （HVs）, 16 gynaecological benign （GB） cases, 18 gynaecological malignant （GM） tumor cases and 16 chronic renal failure （CRF） patients were tested for IL-2 mRNA and IL-4 mRNA by FQ-RT-PCR. The house-keeping gene β-actin was used as the internal control gene of the experiment. The standard curve for log concentration of series of quantitative templates vs threshold cycle （CT） was established by linear regression, and the linear range was 102-107 copies/μl. The imprecision test showed the CV of inter-assay and intra-assay of a high cont- ent sample by FQ-RT-PCR were 7.8% and 12.5%, respectively. The CV of inter-assay and intra-assay of a low content sample were 10.8% and 19.5%, respectively. The IL-2 mRNA expressions in Th of the patients with gynaecological malignant tumor （compared with the HVs and the patients with gynaecological benign disease） and in Th of the CRF patients （compared with the HVs） were declined significantly and at the same time the IL-4 mRNA expression increased significantly （ P 〈 0. 001 ）. A simple, sensitive and accurate FQ-RT-PCR method for the quantitative detection of IL-2 mRNA and IL-4 mRNA has been established. The IL-2 mRNA and IL-4 mRNA expressions in Th cells of the patients with gynaecological malignant tumor and the CRF patients were polarized and displayed Th2 response. It suggests that the function of the Th cells of the patients with gynaecological malignant tumor or CRF is at unbalance.展开更多
BACKGROUND： Previous studies have indicated preventive acupuncture and moxibustion at the Guanyuan point have positive effects on the immune system of menopausal rats. OBJECTIVE： To observe the effects of preventive...BACKGROUND： Previous studies have indicated preventive acupuncture and moxibustion at the Guanyuan point have positive effects on the immune system of menopausal rats. OBJECTIVE： To observe the effects of preventive acupuncture or moxibustion at the Guanyuan point on interleukin-2 （IL-2） and its receptor （IL-2R mRNA） levels in hypothalamus, serum, and uterus of menopausal rats. DESIGN： Randomized control animal experiment. SETTING： School of Acupuncture and Moxibustion, Beijing University of Chinese Medicine. MATERIALS： This study was performed at the Laboratory of Beijing University of Chinese Medicine from November 2004 to July 2005. The vaginal smear method was used to determine the estrus cycle of 9.5-month-old Sprague Dawley （SD） female rats. Three continuous estrus cycles were measured, and 160 rats with estrus cycle disorder, weighing （300 _＋ 20） g, were included in this study. The rats were randomly divided into four groups： preventive acupuncture group （n =48）, preventive moxibustion group （n =48）, model groups （n =48）, and 10-month-old group （n =16）. Sixteen 3.5-month-old healthy SD female rats, weighing （240 ± 20） g, were also assigned to the young group. All rats were clean grade and were purchased from the Laboratory Animal Center, Beijing University of Chinese Medicine. The experiment was in accordance with animal ethics standards. METHODS： Acupuncture was applied to the preventive acupuncture group at 10 months of age. The needle was inserted upward and perpendicularly two fen to a point that was two fen below the Guanyuan point. The rats freely moved around during needle retention （20 minutes）. Acupuncture was administered twice a week and continued for eight weeks. A moxa cone （small） moxibustion was applied to the preventive moxibustion group at 10 months of age. The rats were caught gently and kept at the same position as the preventive acupuncture group. The moxa cone was ignited and held to the Guanyuan point directly. Only one moxa cone was used each time. After moxibustion, the local skin turned slightly red. Moxibustion was used twice a week and continued for eight weeks. Nothing except gently catching and lying was administered to the model group and the young group. MAIN OUTCOME MEASURES： Radioimmunoassay, in situ hybridization, and similar methods were employed to detect IL-2 in serum and the uterus, as well as IL-2R mRNA expression in the hypothalamus and uterus arcuate nuclei, of the preventive acupuncture group, preventive moxibustion group, model groups （at 12, 14, and 16 months）, 10 month old group （at 10 months）, and the young group （at 4 months）. RESULTS： All 176 rats were included in the final analysis. （1） IL-2R mRNA： IL-2R mRNA expression decreased significantly in the uterus of 12-, 14-, and 16-month-old model groups and in the hypothalamus of 14- and 16-month-old model groups （P 〈 0.05-0.01）. Compared to the same month-aged model groups, the expression of IL-2R mRNA increased significantly in the hypothalamus and uterus of 16-month-old preventive acupuncture or moxibustion groups （P 〈 0.05-0.01）.（2） IL-2：IL-2 in serum of 12- and 14-month-old model groups and in the uterus of 12-, 14-, and 16-month-old model groups decreased significantly compared to the young group （P 〈 0.05-0.01）. IL-2 serum levels increased significantly in the 16-month-old group with preventive moxibustion; in addition, IL-2 expression levels increased significantly in the uterus of the 12-month-old group with preventive moxibustion, as well as the 12- and 16-month-old groups with preventive acupuncture （P 〈 0.05-0.01）. CONCLUSION： As a rat ages and becomes menopausal, IL-2R mRNA expression in the hypothalamus and uterus, as well as IL-2 levels in the serum and uterus, decrease. However, preventive acupuncture and preventive moxibustion can repress the decrease to some extent. This effect is even greater in the late stages of menopause （16-month-old rats）.展开更多
基金This work was supported by the National Natural Science Foundation of China(30070566).
文摘In this study, the IL-2 mRNA levels of T lymphocytes in normal mice stimulated by nine Chinese herbal medicinal ingredients （CHMIs） were measured using semiquantification reverse transcription polymerase chain reaction. The results showed that astragalus polysaccharide （APS）, epimedium polysaccharide （EPS）, Chinese angelica polysaccharide （CAPS）, propolis flavone （PF）, and astrogalosides （AS） promoted IL-2 mRNA levels in T lymphocytes in vitro and in vivo to differing degrees, and the level of IL-2 mRNA induced by propolis polysaccharide （PPS） in vitro was higher than that induced by the control, which differed from that of PPS in vivo.
文摘The purpose of the study is to establish a fluorescence quantitative reverse transcription polymerase chain response （FQ-RT-PCR） method for the quantitative determination of IL-2 mRNA and IL-4 mRNA in Th cells, with which the Th cells status of the patients with gynaecological tumors and chronic renal failure （CRF） can be analyzed. IL-2 eDNA and IL-4 eDNA were prepared, and the plasmid pMD18 carrying IL-2 eDNA or IL-4 eDNA fragment was constructed and cloned as the template for quantitative determination. The primers and probes labelled with 6-carbexy-fluorescein （FAM） and 6-carboxy- tetramethylrhodamine （TAMRA） were prepared, and the experimental conditions were optimized to set up the FQ-RT-PCR method for quantitative determination of IL-2 mRNA and IL-4 mRNA. Th cells enriched from peripheral blood mononuclear cells （PBMCs） of 20 healthy volunteers （HVs）, 16 gynaecological benign （GB） cases, 18 gynaecological malignant （GM） tumor cases and 16 chronic renal failure （CRF） patients were tested for IL-2 mRNA and IL-4 mRNA by FQ-RT-PCR. The house-keeping gene β-actin was used as the internal control gene of the experiment. The standard curve for log concentration of series of quantitative templates vs threshold cycle （CT） was established by linear regression, and the linear range was 102-107 copies/μl. The imprecision test showed the CV of inter-assay and intra-assay of a high cont- ent sample by FQ-RT-PCR were 7.8% and 12.5%, respectively. The CV of inter-assay and intra-assay of a low content sample were 10.8% and 19.5%, respectively. The IL-2 mRNA expressions in Th of the patients with gynaecological malignant tumor （compared with the HVs and the patients with gynaecological benign disease） and in Th of the CRF patients （compared with the HVs） were declined significantly and at the same time the IL-4 mRNA expression increased significantly （ P 〈 0. 001 ）. A simple, sensitive and accurate FQ-RT-PCR method for the quantitative detection of IL-2 mRNA and IL-4 mRNA has been established. The IL-2 mRNA and IL-4 mRNA expressions in Th cells of the patients with gynaecological malignant tumor and the CRF patients were polarized and displayed Th2 response. It suggests that the function of the Th cells of the patients with gynaecological malignant tumor or CRF is at unbalance.
基金the National Nature Science Foundation of China, No. 90209026Natural Science Foundation of Beijing, No. 7052037
文摘BACKGROUND： Previous studies have indicated preventive acupuncture and moxibustion at the Guanyuan point have positive effects on the immune system of menopausal rats. OBJECTIVE： To observe the effects of preventive acupuncture or moxibustion at the Guanyuan point on interleukin-2 （IL-2） and its receptor （IL-2R mRNA） levels in hypothalamus, serum, and uterus of menopausal rats. DESIGN： Randomized control animal experiment. SETTING： School of Acupuncture and Moxibustion, Beijing University of Chinese Medicine. MATERIALS： This study was performed at the Laboratory of Beijing University of Chinese Medicine from November 2004 to July 2005. The vaginal smear method was used to determine the estrus cycle of 9.5-month-old Sprague Dawley （SD） female rats. Three continuous estrus cycles were measured, and 160 rats with estrus cycle disorder, weighing （300 _＋ 20） g, were included in this study. The rats were randomly divided into four groups： preventive acupuncture group （n =48）, preventive moxibustion group （n =48）, model groups （n =48）, and 10-month-old group （n =16）. Sixteen 3.5-month-old healthy SD female rats, weighing （240 ± 20） g, were also assigned to the young group. All rats were clean grade and were purchased from the Laboratory Animal Center, Beijing University of Chinese Medicine. The experiment was in accordance with animal ethics standards. METHODS： Acupuncture was applied to the preventive acupuncture group at 10 months of age. The needle was inserted upward and perpendicularly two fen to a point that was two fen below the Guanyuan point. The rats freely moved around during needle retention （20 minutes）. Acupuncture was administered twice a week and continued for eight weeks. A moxa cone （small） moxibustion was applied to the preventive moxibustion group at 10 months of age. The rats were caught gently and kept at the same position as the preventive acupuncture group. The moxa cone was ignited and held to the Guanyuan point directly. Only one moxa cone was used each time. After moxibustion, the local skin turned slightly red. Moxibustion was used twice a week and continued for eight weeks. Nothing except gently catching and lying was administered to the model group and the young group. MAIN OUTCOME MEASURES： Radioimmunoassay, in situ hybridization, and similar methods were employed to detect IL-2 in serum and the uterus, as well as IL-2R mRNA expression in the hypothalamus and uterus arcuate nuclei, of the preventive acupuncture group, preventive moxibustion group, model groups （at 12, 14, and 16 months）, 10 month old group （at 10 months）, and the young group （at 4 months）. RESULTS： All 176 rats were included in the final analysis. （1） IL-2R mRNA： IL-2R mRNA expression decreased significantly in the uterus of 12-, 14-, and 16-month-old model groups and in the hypothalamus of 14- and 16-month-old model groups （P 〈 0.05-0.01）. Compared to the same month-aged model groups, the expression of IL-2R mRNA increased significantly in the hypothalamus and uterus of 16-month-old preventive acupuncture or moxibustion groups （P 〈 0.05-0.01）.（2） IL-2：IL-2 in serum of 12- and 14-month-old model groups and in the uterus of 12-, 14-, and 16-month-old model groups decreased significantly compared to the young group （P 〈 0.05-0.01）. IL-2 serum levels increased significantly in the 16-month-old group with preventive moxibustion; in addition, IL-2 expression levels increased significantly in the uterus of the 12-month-old group with preventive moxibustion, as well as the 12- and 16-month-old groups with preventive acupuncture （P 〈 0.05-0.01）. CONCLUSION： As a rat ages and becomes menopausal, IL-2R mRNA expression in the hypothalamus and uterus, as well as IL-2 levels in the serum and uterus, decrease. However, preventive acupuncture and preventive moxibustion can repress the decrease to some extent. This effect is even greater in the late stages of menopause （16-month-old rats）.
