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Objective@#To investigate the inhibition of tanshinoneⅡA (TanⅡA) on the proliferation of lung cancer cells.@*Methods@#Human lung cancer cell lines A549, SK-MES-1, H446 and H460 were cultured in vitro and treated with TanⅡA at concentrations of 0.3, 0.6, 1.3, 2.5, 5.0, 10.0 mg/mL, while untreated cells served as controls. Cell proliferation was measured by using CCK-8 assay, and apoptosis was measured using flow cytometry, while the expression of apoptosis-related proteins was determined using Western blotting. The apoptotic rate of lung cancer cells was compared between Tan ⅡA-treated cells and untreated cells.@*Results@# Tan ⅡA inhibited lung cancer cell proliferation in a time-dependent and concentration-dependent manner, and the survival rates of A549, SK-MS-1, H446 and H460 cells reduced with the concentration of TanⅡA (t=4.503, 2.114, 2.103 and 3.567; all P<0.05) and the duration of TanⅡA treatment (t=5.189, 3.079, 3.023 and 3.845; all P<0.05). The 48 h half maximal inhibitory concentrations (IC50 values) of TanⅡA were (1.18±0.12), (0.78±0.08), (1.55±0.16) and (1.27±0.14) mg/mL against A549, SK-MES-1, H446 and H460 cells, respectively. Following 48 h treatment with Tan ⅡA at a concentration of 2.5 mg/mL, the apoptotic rates of A549, SK-MS-1, H446 and H460 cells were (34.97±3.78)%, (37.62±2.48)%, (18.27±2.98)% and (19.17±2.30)%, which were significantly significantly higher than those of untreated cells [(4.86±0.36) %, (3.21±0.48) %, (3.25±0.26)% and (2.66±0.19)%, all P<0.05]. Reduced Akt1 protein expression, elevated Bax/Bcl-2 expression ratio, and elevated caspase-9 and caspase-3 protein expression were detected in lung cancer cells treated with 2.5 mg/mL TanⅡA for 48 h relative to untreated cells@*Conclusion@#TanⅡA may inhibit lung cancer cell proliferation in a time- and concentration-dependent manner via the Bax/Bcl-2/caspase-9/caspase-3 pathway.
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Objective To know the Tanshinone Ⅱ A induce human small cell lung cancer cells (H446 cells) apoptosis,and discuss the possible molecular mechanism.Methods Determined by MTT method of different concentrations of Tanshinone Ⅱ A impact on human small cell lung cancer H446 cells proliferation;Determined by Hoechst 33258 method of different concentrations Tanshinone Ⅱ A influence on human small cell lung cancer H446 cells apoptosis;Determined by qPCR method of different concentrations of Tanshinone Ⅱ A impact on human small cell lung cancer H446 cells apoptosis gene;Determined by Westen blot method of different concentrations of Tanshinone lⅡ A effect on lung cancer H446 cells apoptosis.Results Different drug concentrations of Tanshinone Ⅱ A (0.3,0.6,1.25,2.5,5 and 10μg/ml) inhibit human small cell lung cancer H446 cells proliferation and depend by the concentration and time;With the increase of the concentration of the drug in the formation of human small cell lung cancer H446 cells apoptosis corpuscle number increase,significant difference was found in high concentration group (P ＜ 0.05);High concentration group of Tanshinone Ⅱ A promote apoptosis gene (Bax,caspase-9,caspase-3) expression,and inhibit apoptosis inhibiting genes (Akt,the Bcl-2) expression and there were significant differences (P ＜ 0.05);High concentration group of Tanshinone Ⅱ A promoting apoptosis protein (Bax,caspase-9,caspase-3)expression,and inhibit apoptosis inhibiting protein (Akt,the Bcl-2) expression and there were significant differences (P ＜ 0.05).Conclusion Tanshinone Ⅱ A may be through Akt-Bax/Bcl-2-caspase-9-caspase-3 signaling pathways induced human small cell lung cancer H446 cells apoptosis.
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Objective To detect the changes of microbial community functional diversity of corpses with different postmortem interval(PMI)and to evaluate forensic application value for estimating PMI. Methods The cultivation of microbial community from the anal swabs of aSusscrofaand a human corpse placed in field environment from 0 to 240 h after death was performed using the Biolog-Eco Mi-croplate and the variations of the absorbance values were also monitored. Combined with the technology of forensic pathology and flies succession, the metabolic characteristics and changes of microbial commu-nity on the decomposed corpse under natural environment were also observed.Results The diversity of microbial metabolism function was found to be negatively correlated with the number of maggots in the corpses. The freezing processing had the greatest impact on average well color development value at 0 h and the impact almost disappeared after 48 h. The diversity of microbial metabolism of the samples be-came relatively unstable after 192 h. The principal component analysis showed that 31 carbon sources could be consolidated for 5 principal components(accumulative contribution ratio >90%). The carbon source tsquare-analysis showed thatN-acetyl-D-glucosamine andL-serine were the dominant carbon sources for estimating the PMI(0=240 h)of theSusscrofaand human corpse.Conclusion The Biolog-Eco method can be used to reveal the metabolic differences of the carbon resources utilization of the microbial community on the corpses during 0-240 h after death, which could provide a new basis for estimating the PMI.
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Objective To explore the growing development and community succession of main sarcos-aphagous insects on pig carcasses in summer indoor and outdoor environment in Shenzhen area and to estimate the postmortem interval (PMI). Methods From early May to August in 2013, in Forensic Med-ical Examination Center of Shenzhen Public Security Bureau, the main insect species and the decomposi-tion process were observed in two adult pig carcasses of simulative indoor and outdoor environment. The different decomposition stages and the community succession of insects were recorded. Results The indoor and outdoor pig carcasses showed skeleton 412.5 and 325 hours after death, respectively. The main species of flies on pig carcasses were Chrysomya megacephala, Chrysomya rufifacies and Chrysomya chani. The main species of beetles were Crecphilus maxillosus, Necrobia ruficollis, Saprinus splendens and Dermestes maculatu. The dominant species of flies in the outdoor pig carcasses obviously produced the second generations due to the effect of mass rainfall, nor in the indoor pig carcasses. Conclusion There are regular patterns on the community succession of insects on pig carcasses in summer indoor and out-door environment in Shenzhen area. The activity patterns of seven typical insects and their larva show important value for estimating PMI.
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Objective To detect the expressions of microRNA-126 (miR-126) and microRNA-7 (miR-7) in esophageal squamous cell carcinoma (ESCC) and to analyze their correlations with clinicopathologic features and prognosis of ESCC.Methods The expressions of miR-126 and miR-7 in 116 ESCC samples and matched normal tissue samples were detected by real-time PCR.Statistical analysis was used to find the relationships among the expressions of miR-126 and miR-7,pathological characteristics and prognosis.Results Low expression,normal expression and high expression of miR-126 were found in 73 (62.9％),35 (30.2％) and 8 (6.9％) carcinoma samples respectively.Low expression,normal expression and high expression of miR-7 were found in 52 (44.8％),35 (30.2％) and 29 (25.0％) carcinoma samples respectively.The disease-free survival in patients with low expression of miR-126 and miR-7 was shorter than that in patients with non-low expression (x2 =4.268,P ＜0.05 ; x2 =4.993,P ＜0.05).The low expression of miR-126 was correlated with tumor location,family history and drinking (x2 =14.564,P ＜ 0.05 ; x2 =5.691,P ＜ 0.05 ; x2 =4.971,P ＜ 0.05),but was uncorrelated with gender,age,diferentiation,infiltration,lymphatic metastasis and smoking (all P ＞ 0.05).The low expression of miR-7 was uncorrelated with pathological characteristics of ESCC (all P ＞ 0.05).Conclusion The low expressions of miR-126 and miR-7 may be related to the prognosis of patients with ESCC,and have a certain clinical detection significance.