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Objective: To investigate the inhibitory effects of ampelopsin (AMP) on proliferation, autophagy and apoptosis to human colon cancer cells and its possible mechanism. To observe the function of autophagy in the apoptosis of colon cancer cells as well. Methods: The lentivirus infection method was adopted here to construct double fluorescence [green fluorescent protein (GFP) and red fluorescent protein (RFP)] labeled microtubule-associated protein light chain 3 (LC3) overexpressed recombinant HCT116-RFP-GFP-LC3 cells. The GFP and RFP labeled LC3 expression in HCT116-RFP-GFP-LC3 cells were observed by fluorescence microscope, and Western blotting was used to detect the expression of LC3 in HCT116-RFP-GFP-LC3 cells at the same time. The proliferation inhibition of recombination HCT116-RFP-GFP-LC3 cells and parental HCT116 cells treated with different concentrations of AMP (0, 50, 100, 175, 250, 350, 400 and 450 μg/mL) was detected by CCK-8 assay and the cell morphology was observed. The apoptotic rate of HCT116 cells treated with AMP (0, 25 50, and 75 μg/mL) was detected by FCM method, and the apoptotic-related proteins Bax, nuclear factor-kappa B (NF-κB) and Bcl-2 expression levels were detected by Western blotting. Autophagy induction was detected by counting fluorescence dots triggered by transformation of LC3to LC3Ⅱ visualized by laser confocal microscopy after HCT116-RFP-GFP-LC3 cells treated with AMP (0, 25 50, and 75 μg/mL) alone or combined with bafilomycin A1(Baf A1) (an autophagy inhibitor) (20 nmol/L). After the HCT116 cells were treated with AMP (25 μg/mL) alone or combined with Baf A1 (20 nmol/L), the apoptotic rate was detected by FCM method, and the expression level of autophagy-related proteins LC3Ⅱ and p62 as well as apoptotic-related proteins were detected by Western blotting. Results: RFP-GFP-LC3 could stably express in HCT116-RFP-GFP-LC3 cells, indicating that HCT116-RFP-GFP-LC3 recombinant cells were successfully constructed. The results of CCK-8 assay suggested that AMP could significantly inhibit the proliferation of recombinant and parental cells in a concentration-dependent manner. Meanwhile, there was no significant difference in cell morphology or cell proliferation inhibition rate between these two cells (P > 0.05). The apoptosis rate of HCT116 cells increased remarkably along with the enhancement of AMP concentration which means there is a concentration-dependent manner (all P < 0.05). The expression levels of pro-apoptotic protein Bax and NF-κB were increased, and the apoptosis inhibitory protein Bcl-2 was down-regulated (all P < 0.01). The confocal microscopy results showed that with the increase of AMP concentration, the numbers of autophagy points were significantly increased (P < 0.01), and the expression level of autophagy protein LC3Ⅱ was up-regulated, indicating that AMP could trigger autophagy on colon cancer cells. Comparing with AMP alone group, AMP combined with Baf A1 could increase the apoptosis rate of HCT116 significantly (P < 0.05), and the expression level of apoptotic protein Bax promoted remarkably (P < 0.05). The expression level of the apoptosis inhibitory protein Bcl-2 demoted as well. All these results suggest that autophagy blocking could promote AMP induced apoptosis. Conclusion: AMP can inhibit the proliferation of colon cancer cells HCT116, inducing autophagy and apoptosis; inhibition of autophagy can promote AMP-induced apoptosis on colon cancer cells.
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Objective To investigate the clinical efficacy and safety of Xiao'er Chiqiaoqingre granule in the treatment of acute upper respiratory tract infection.Methods From January 2017 to December 2017,a total of 116 children with upper respiratory tract infection in Taizhou Integrated Traditional Chinese and Western Medicine Hospital were selected and randomly divided into control group and observation group according to different treatment regimens,with 58 cases in each group.The control group was given ribavirin granules,ibuprofen suspension drops for treatment.The observation group was given Xiao'er Chiqiaoqingre granule on the basis of the control group.The two groups were treated for 5 days,then the clinical effect,the improvement of clinical symptoms,the time of cure,the level of serum cytokines and safety were compared between the two groups.Results After treatment,the cure rate of the observation group was 96.6％,which was significantly higher than 81.0％ of the control group (P ＜0.05).The antipyretic time,cough disappeared time,healing time in the observation group were (1.4 ± 0.5) d,(2.1 ± 0.4) d,(4.5 ± 1.4) d,respectively,which were significantly shorter than those in the control group[(2.6 ± 0.9) d,(3.4 ± 1.1) d,(5.8 ±1.9) d] (all P ＜ 0.05).The throat irritation subsided time of the observation group was (3.5 ± 1.1) d,which of the control group was (3.8 ± 1.3) d,the difference was not statistically significant (P ＞ 0.05).The serum levels of IL-6,IL -10 and TNF-in the observation group were (108.45 ± 25.61) μg/L,(34.88 ± 9.07) μg/L,(1.26 ± 0.86) mg/mL,respectively,which were significantly lower than those in the control group[(129.27 ± 28.31) μg/L,(43.27 ±10.09)μg/L,(2.11 ± 1.03)mg/mL] (all P ＜ 0.05).There were no other serious adverse reactions in the two groups.Conclusion Xiao'er Chiqiaoqingre granule in the treatment of acute upper respiratory tract infection can significantly improve clinical symptoms,improve clinical curative effect,and has good safety and certain clinical value.
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Objective To investigate the effect of Azithromycin combined with terbutaline on the treatment of pneumonia in children and its effect on symptom improvement.MethodsThe clinical data of children with pneumonia treated in Taizhou Hospital of traditional Chinese and Western medicine from January 2014 to December 2016 were retrospectively analyzed, according to the treatment methods,the patients were divided into control group and observation group, the control group was given azithromycin treatment, the observation group on this basis to give terbutaline atomization treatment.The therapeutic effects of the two groups were observed, and the differences of clinical symptom score, inflammatory factor level were compared between the two groups before and after treatment.ResultsThe effective rate of the observation group was 98%, which was significantly higher than that of the control group (84%);The clinical symptom score had no difference between the two groups before treatment, after treatment, the observation group of cough, expectoration, fever and rales were lower than those in the control group;The two groups had no difference between the levels of proinflammatory cytokines, after treatment, the observation group IL-6, IL-8, TNF-and CRP levels were lower than the control group.ConclusionAzithromycin combined with terbutaline in the treatment of children with pneumonia has a better therapeutic effect, can significantly improve the clinical symptoms, reduce the level of inflammatory factors, has a good clinical value.
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<p><b>OBJECTIVE</b>To study the effects and mechanism of lovastatin on cell proliferation and expression of proinflammatory cytokines in cultured human glomerular mesangial cells.</p><p><b>METHODS</b>The influence of lovastatin on HMC proliferation was evaluated with 3H-thymidine incorporation. mRNA expression of proinflammatory cytokines (IL-1 beta, IL-6, TNF-alpha, and MCP-1) and activation of NF-kappa B of HMC were measured using Reverse transcription-polymerase chain reaction (RT-PCR) and electrophoretic mobility shift assay (EMSA) respectively.</p><p><b>RESULTS</b>Lovastatin was found to have inhibitory effects on human mesangial cell (HMC) proliferation and lipopolysaccharide (LPS)-mediated human mesangine cell HMC mRNA expression of proinflammatory cytokines via activation of NF-kappa B. The effect of lovstatin on HMC could be prevented when the mevalonate and farnesol were added to the culture.</p><p><b>CONCLUSION</b>Lovastatin may decrease HMC proliferation and production of proinflammatory cytokines through the inhibition of NF-kappa B activation. This provided experimental evidence for further evaluation of the renal protective effect of HRI, suggesting that it may be a potent agent for prevention of progressive renal diseases aside from its lipid-lowering effect.</p>