Objective To study the modulation of interleukin 6 (IL 6) generation from human thyroid cells. MethodsThyroid tissues were obtained at surgery from patients with Graves disease. IL 6 in the supernatant of cultured...Objective To study the modulation of interleukin 6 (IL 6) generation from human thyroid cells. MethodsThyroid tissues were obtained at surgery from patients with Graves disease. IL 6 in the supernatant of cultured thyrocytes was detected with ultra sensitive ELISA. mRNA was extracted respectively from primary human cultured thyroidal cells under stimulated and unstimulated conditions,and semiquantitative reverse transcriptase polymerase chain reaction (RT PCR) technique was used for IL 6 mRNA detection. Results① In the basic conditions,thyroid cells can produce high concentration IL 6. Thyrotropin (TSH,10 3mU/L), tumor necrosis factor α(TNF α,10 -1 U/ml),interleukin 1(IL 1,10 U/ml) and adrenaline(10 -5 mol/L) significantly increased IL 6 levels in the supernatant of cultured thyroid cells,whereas dexamethasone (10 mmol/L) markedly suppressed IL 6 release from thyrocytes. NaI of 10 -4 mol/L exerted no effect on the production of IL 6. ② Compared with the basal results,at a concentration of 10-10 3 mmol/L,dexamethasone could gradually suppress IL 6 gene expression on human thyrocytes,while TNF α could obviously stimulate the production of IL 6 mRNA in the range of 10 -1 -10 2 U/ml. On the level of 10 2 U/ml,IL 1 increased the expression of IL 6,but the same effect could not be shown when IL 1 was 10 -1 or 10 U/ml. Sodium iodine (NaI) could not affect IL 6 gene from the level of 10 -8 to 10 -4 mol/L. ConclusionIL 6 produced by thyrocytes might be regulated by many factors that modulate thyroid functions in vitro as well as in vivo.展开更多
Analysis of proliferative activity and the ability to neuron differentiation was performed in cultured cells of the brain and spinal cord of juvenile masu salmon Oncorhynchus masou. Proliferating cell nuclear antigen ...Analysis of proliferative activity and the ability to neuron differentiation was performed in cultured cells of the brain and spinal cord of juvenile masu salmon Oncorhynchus masou. Proliferating cell nuclear antigen (PCNA) was used as a proliferative marker, while the markers of neuronal differentiation—a neuron protein HuCD, and a neuron-specific transcriptional factor with two DNA- binding sites Pax6—detected neurons. The results showed that cell proliferation occurred mainly in the suspension cell fraction. In monolayer, a few cells were only found to express PCNA. The results of morphological and immunohistochemical analysis allow us to conclude that proliferative activity in primary cultures from the O. masou brain is mainly connected with the suspension fraction of small cells. In contrast, a positive correlation between the cells expressing cystathionine β-synthase (CBS), a marker of H2S synthesis, and the cells expressing PCNA in the monolayer, indicates the participation of H2S in proliferative activity of neurons in primary cultures. The data obtained suggest that the hydrogen sulphide is also involved in the process of differentiation.展开更多
BACKGROUND： Pseudomonas aeruginosa mannose-sensitive hemagglutinin （PA-MSHA） parenteral injection is used as a broad-spectrum immunomodulator. It remains unclear whether PA-MSHA exhibits inhibitory effects on tumor...BACKGROUND： Pseudomonas aeruginosa mannose-sensitive hemagglutinin （PA-MSHA） parenteral injection is used as a broad-spectrum immunomodulator. It remains unclear whether PA-MSHA exhibits inhibitory effects on tumor cell growth. OBJECTIVE： To investigate inhibitory mechanisms of PA-MSHA-induced proliferation in rat C6 glioma cells in vitro. DESIGN, TIME AND SETTING： Comparative observation and in vitro experiments were performed at the Key Laboratory of Natural Medicine, Kunming Medical College, China from July 2008 to April 2009. MATERIALS： Rat C6 glioma cell line （Shanghai Institute of Cell Biology, Chinese Academy of Sciences, China） and PA-MSHA parenteral injection （Beijing Wanteer Bio-Pharmaceutical, China） were used in the present study. METHODS： Rat C6 glioma cells in logarithmic growth phase were harvested in vitro. Adherent monolayer cells were respectively treated with PA-MSHA at final colony-forming units （cfu） of 1 ×10^8 cfu/mL, 2 × 10^8 cfu/mL, 4 × 10^8 cfu/mL, 6 × 10^8 cfu/mL, and 8 ×10^8 cfu/mL following 24 hours of conventional culture. MAIN OUTCOME MEASURES： MTT colorimetric assay was utilized to determine the inhibitory rate of C6 glioma cells following treatment with various concentrations of PA-MSHA at different times. Cell apoptosis was detected by fluorescent microscopy following Hoechst 33258 staining. Flow cytometry was used to measure PA-MSHA effects on C6 cell cycle. RESULTS： Inhibitory rate of C6 glioma cells increased with prolonged time and increased dose. Hoechst 33258 staining revealed obvious morphological changes in apoptotic C6 glioma cells. Flow cytometry revealed hypodiploid peaks, Le., apoptotic peak, and the apoptotic rate in cells during S-phase significantly increased with increased concentrations in the experimental groups. CONCLUSION： With in vitro experiments, PA-MSHA preparations inhibited C6 glioma cell proliferation in a time- and dose-dependent manner. These mechanisms are likely associated with cell apoptosis induction and inhibition of the S phase.展开更多
The objectives of this study were to explore whether heparin-binding proteins, separated by fast protein liquid chromatography from boar seminal plasma influence the release of prostaglandins F2α, (PGF2α), E2 (PGE2)...The objectives of this study were to explore whether heparin-binding proteins, separated by fast protein liquid chromatography from boar seminal plasma influence the release of prostaglandins F2α, (PGF2α), E2 (PGE2) and interleukin-6 (IL-6) by porcine endometrial and cervical cells and even bovine endometrial cells. In Experiment I, we showed that release of PGF2α by endometrial epithelial, endometrial stromal and cervical stromal cells to the medium was inhibited (p α (TNFα) stimulated release of IL-6 by endometrial and cervical stromal cells after 24 h incubation, but in the presence of heparin-binding proteins, this stimulation was attenuated. Release of PGF2α by cryopreserved (Experiment II) and primary (Experiment III) cervical stromal cells was significantly inhibited after 3 h incubation with 66 - 95.4 μg of heparin- binding proteins. A significant inhibition of PGE2 release by cryopreserved and primary cervical stromal cells was already achieved after incubation with 16.5 - 23.9 μg of heparin-binding proteins. The release of IL-6 by cryopreserved cells was stimulated after 3 h incubation with heparin- binding proteins in a dose dependent manner in contrast to the release of IL-6 by freshly isolated cervical stromal cells. We also found (Experiment IV) that porcine heparin-binding seminal plasma proteins inhibited release of PGF2α and stimulated release of IL-6 by bovine endometrial epithelial cells. In conclusion, a group of heparin-binding proteins separated by fast protein liquid chromatography from boar seminal plasma inhibit PGF2α, PGE2 and stimulate IL-6 release by porcine endometrial and cervical cells and even by bovine endometrial cells. Thus, these proteins have a similar effect as the entire seminal plasma.展开更多
基金This paper is supported by Jiang su Province Natural Science Fund (BK95080314 )
文摘Objective To study the modulation of interleukin 6 (IL 6) generation from human thyroid cells. MethodsThyroid tissues were obtained at surgery from patients with Graves disease. IL 6 in the supernatant of cultured thyrocytes was detected with ultra sensitive ELISA. mRNA was extracted respectively from primary human cultured thyroidal cells under stimulated and unstimulated conditions,and semiquantitative reverse transcriptase polymerase chain reaction (RT PCR) technique was used for IL 6 mRNA detection. Results① In the basic conditions,thyroid cells can produce high concentration IL 6. Thyrotropin (TSH,10 3mU/L), tumor necrosis factor α(TNF α,10 -1 U/ml),interleukin 1(IL 1,10 U/ml) and adrenaline(10 -5 mol/L) significantly increased IL 6 levels in the supernatant of cultured thyroid cells,whereas dexamethasone (10 mmol/L) markedly suppressed IL 6 release from thyrocytes. NaI of 10 -4 mol/L exerted no effect on the production of IL 6. ② Compared with the basal results,at a concentration of 10-10 3 mmol/L,dexamethasone could gradually suppress IL 6 gene expression on human thyrocytes,while TNF α could obviously stimulate the production of IL 6 mRNA in the range of 10 -1 -10 2 U/ml. On the level of 10 2 U/ml,IL 1 increased the expression of IL 6,but the same effect could not be shown when IL 1 was 10 -1 or 10 U/ml. Sodium iodine (NaI) could not affect IL 6 gene from the level of 10 -8 to 10 -4 mol/L. ConclusionIL 6 produced by thyrocytes might be regulated by many factors that modulate thyroid functions in vitro as well as in vivo.
文摘Analysis of proliferative activity and the ability to neuron differentiation was performed in cultured cells of the brain and spinal cord of juvenile masu salmon Oncorhynchus masou. Proliferating cell nuclear antigen (PCNA) was used as a proliferative marker, while the markers of neuronal differentiation—a neuron protein HuCD, and a neuron-specific transcriptional factor with two DNA- binding sites Pax6—detected neurons. The results showed that cell proliferation occurred mainly in the suspension cell fraction. In monolayer, a few cells were only found to express PCNA. The results of morphological and immunohistochemical analysis allow us to conclude that proliferative activity in primary cultures from the O. masou brain is mainly connected with the suspension fraction of small cells. In contrast, a positive correlation between the cells expressing cystathionine β-synthase (CBS), a marker of H2S synthesis, and the cells expressing PCNA in the monolayer, indicates the participation of H2S in proliferative activity of neurons in primary cultures. The data obtained suggest that the hydrogen sulphide is also involved in the process of differentiation.
文摘BACKGROUND： Pseudomonas aeruginosa mannose-sensitive hemagglutinin （PA-MSHA） parenteral injection is used as a broad-spectrum immunomodulator. It remains unclear whether PA-MSHA exhibits inhibitory effects on tumor cell growth. OBJECTIVE： To investigate inhibitory mechanisms of PA-MSHA-induced proliferation in rat C6 glioma cells in vitro. DESIGN, TIME AND SETTING： Comparative observation and in vitro experiments were performed at the Key Laboratory of Natural Medicine, Kunming Medical College, China from July 2008 to April 2009. MATERIALS： Rat C6 glioma cell line （Shanghai Institute of Cell Biology, Chinese Academy of Sciences, China） and PA-MSHA parenteral injection （Beijing Wanteer Bio-Pharmaceutical, China） were used in the present study. METHODS： Rat C6 glioma cells in logarithmic growth phase were harvested in vitro. Adherent monolayer cells were respectively treated with PA-MSHA at final colony-forming units （cfu） of 1 ×10^8 cfu/mL, 2 × 10^8 cfu/mL, 4 × 10^8 cfu/mL, 6 × 10^8 cfu/mL, and 8 ×10^8 cfu/mL following 24 hours of conventional culture. MAIN OUTCOME MEASURES： MTT colorimetric assay was utilized to determine the inhibitory rate of C6 glioma cells following treatment with various concentrations of PA-MSHA at different times. Cell apoptosis was detected by fluorescent microscopy following Hoechst 33258 staining. Flow cytometry was used to measure PA-MSHA effects on C6 cell cycle. RESULTS： Inhibitory rate of C6 glioma cells increased with prolonged time and increased dose. Hoechst 33258 staining revealed obvious morphological changes in apoptotic C6 glioma cells. Flow cytometry revealed hypodiploid peaks, Le., apoptotic peak, and the apoptotic rate in cells during S-phase significantly increased with increased concentrations in the experimental groups. CONCLUSION： With in vitro experiments, PA-MSHA preparations inhibited C6 glioma cell proliferation in a time- and dose-dependent manner. These mechanisms are likely associated with cell apoptosis induction and inhibition of the S phase.
文摘The objectives of this study were to explore whether heparin-binding proteins, separated by fast protein liquid chromatography from boar seminal plasma influence the release of prostaglandins F2α, (PGF2α), E2 (PGE2) and interleukin-6 (IL-6) by porcine endometrial and cervical cells and even bovine endometrial cells. In Experiment I, we showed that release of PGF2α by endometrial epithelial, endometrial stromal and cervical stromal cells to the medium was inhibited (p α (TNFα) stimulated release of IL-6 by endometrial and cervical stromal cells after 24 h incubation, but in the presence of heparin-binding proteins, this stimulation was attenuated. Release of PGF2α by cryopreserved (Experiment II) and primary (Experiment III) cervical stromal cells was significantly inhibited after 3 h incubation with 66 - 95.4 μg of heparin- binding proteins. A significant inhibition of PGE2 release by cryopreserved and primary cervical stromal cells was already achieved after incubation with 16.5 - 23.9 μg of heparin-binding proteins. The release of IL-6 by cryopreserved cells was stimulated after 3 h incubation with heparin- binding proteins in a dose dependent manner in contrast to the release of IL-6 by freshly isolated cervical stromal cells. We also found (Experiment IV) that porcine heparin-binding seminal plasma proteins inhibited release of PGF2α and stimulated release of IL-6 by bovine endometrial epithelial cells. In conclusion, a group of heparin-binding proteins separated by fast protein liquid chromatography from boar seminal plasma inhibit PGF2α, PGE2 and stimulate IL-6 release by porcine endometrial and cervical cells and even by bovine endometrial cells. Thus, these proteins have a similar effect as the entire seminal plasma.
