基于数据挖掘技术探讨中医治疗肺癌放疗患者的用药规律

目的 探讨中医治疗肺癌放疗患者的用药规律。方法 检索中国知网数据库（CNKI）、万方数据库、维普数据库（VIP）、中国生物医学文献数据库（CBM）、PubMed数据库公开发表的治疗肺癌放疗患者的中药处方,运用频次分析、聚类分析、关联规则分析方法对收集的处方进行数据挖掘,分析其用药规律。结果 共收集到处方70张,涉及176味中药,使用频次共计890次;使用频次≥9的药物共32味;使用频次前三位的单味中药是黄芪、甘草、麦冬;使用频次前三位的药类是补虚药、清热药、化痰止咳平喘药;对32味药物进行聚类分析和关联规则分析,得出2个有效聚类群和13组药对,以补气药和滋阴药配伍为主,辅以补血活血、化痰止咳药。结论 治疗肺癌放疗患者的中药处方以补益药、清热药和止咳药为主,治法以益气养阴、清肺止咳为主。     

三物白散对Survivin-RNA基因沉默转染胃癌SGC-7901细胞凋亡、迁移的影响

目的 探讨三物白散对Survivin-RNA基因沉默转染胃癌SGC-7901细胞凋亡、迁移的影响。方法 将实验分成6组：空白对照组、LV-RNAi 组、Survivin-RNAi-LV组、空白对照+中药组、LV-RNAi+中药组、Survivin-RNAi-LV+中药组;采用流式细胞仪检测各组细胞的凋亡,划痕实验检测各组细胞的迁移能力。结果 细胞凋亡率：Survivin-RNAi-LV组明显高于空白对照组和LV-RNAi组（P<0.05）;Survivin-RNAi-LV+中药组与空白对照+中药组、LV-RNAi+中药组比较明显较高（P<0.05）;Survivin-RNAi-LV+中药组高于Survivin-RNAi-LV组,差异具有统计学意义（P<0.05）。胃癌细胞的24 h迁移率：Survivin-RNAi-LV组明显低于空白对照组和LV-RNAi组（P<0.05）;Survivin-RNAi-LV+中药组与空白对照+中药组、LV-RNAi+中药组比较,其结果明显低于后两组（P<0.05）;Survivin-RNAi -LV+中药组与Survivin-RNAi-LV组比较,细胞迁移率明显降低,差异有统计学意义（P<0.05）。结论 Survivin基因沉默转染,增强了三物白散诱导胃癌SGC-7901细胞凋亡和抑制其迁移的作用。     

Effect of Linc00152 targeting miR-376c-3p on viability,apoptosis and radiosensitivity of cervical cancer cells

AIM: To investigate the effect of Linc00152 on the viability, apoptosis and radiosensitivity of cervical cancer cells. METHODS: RT-qPCR was used to detect the expression levels of Linc00152 and microRNA-376c-3p(miR-376c-3p) in human cervical cancer HeLa cells and SiHa cells, and normal cervical Ect1/E6E7 cells. The cervical cancer HeLa cells with low Linc00152 expression or miR-376c-3p over-expression were established. MTT assay, flow cytometry, colony formation assay and Western blot were used to determine the cell viability, apoptosis, radiosensitivity and related protein expression. The dual-luciferase reporter assay was used to verify the regulatory relationship between Linc00152 and miR-376c-3p in the HeLa cells. RESULTS: Compared with the Ect1/E6E7 cells, Linc00152 was up-regulated in the HeLa cells and SiHa cells, and miR-376c-3p was down-regulated (P < 0.05). Low expression of Linc00152 or over-expression of miR-376c-3p inhibited the viability of HeLa cells, induced apoptosis, enhanced the radiosensitivity, inhibited the protein expression of cyclin D and Bcl-2, and promoted the protein expression of P21 and Bax (P < 0.05). Linc00152 negatively regulated miR-376c-3p expression in the HeLa cells, and inhibition of miR-376c-3p expression reversed the effect of low expression of Linc00152 on HeLa cell viability, apoptosis and radiosensitivity. CONCLUSION: Linc00152 is highly expressed in the cervical cancer cells. Linc00152 affects the viability, apoptosis and radiosensitivity of HeLa cells by targeting miR-376c-3p, which is a potential diagnosis and treatment target for cervical cancer.     

Reversing effect of naringin on cisplatin resistance in human lung cancer A549/DDP cells

AIM: To investigate the effect of naringin (NRG) on cisplatin (DDP) resistance in human lung cancer A549/DDP cells and its possible mechanism. METHODS: A549/DDP cells were cultured in vitro and treated with NRG and/or DDP at different concentrations for 24 h, and then the cell viability were measured by CCK-8 assay. The combination index (CI) of NRG and DDP were analyzed by Chou-Talalay method. The apoptosis rate was analyzed by flow cytometry. Western blot was performed to detect the protein levels of P-glycoprotein (P-gp), multidrug resistance-associated protein 1 (MRP1), p-Akt, CXC chemokine receptor 4 (CXCR4), cleaved caspase-3, Bcl-2 and Bax.RESULTS: The protein levels of P-gp, MRP1, p-Akt and CXCR4 in the A549/DDP cells were higher than those in the A549 cells (P<0.05). The cell viability was remarkably reduced in a dose-dependent manner when A549/DDP cells were exposed to NRG and/or DDP (P<0.05), and the IC₅₀ values of NRG and DDP were 36.92 μmol/L and 129.77 μmol/L, respectively. When the inhibition rate exceeded 15%, NRG in combination with DDP produced a synergistic effect (CI<1). Combination treatment with NRG and DDP significantly induced apoptosis (P<0.05), up-regulated the protein levels of cleaved caspase-3 and Bax, and down-regulated the protein level of Bcl-2 (P<0.05). Meanwhile, NRG remarkably down-regulated the protein levels of P-gp, MRP1, p-Akt and CXCR4 in a dose-dependent manner (P<0.05). CONCLUSION: NRG may enhance the sensibility of A549/DDP cells to DDP most likely via up-regulating the protein level of Bax and down-regulating the protein levels of Bcl-2, P-gp, MRP1, p-Akt and CXCR4.     

SIRT1 promotes autophagy of pancreatic cancer cells induced by hypoxia via regulating FOXO1/RAB7 signaling pathway

AIM: To investigate the effect of SIRT1 on the autophagy of pancreatic cancer cells under hypoxia condition, and to analyze the underlying mechanism of regulating FOXO1/RAB7 signaling pathway. METHODS: Western blot and immunofluorescence methods were used to determine the expression of SIRT1 in the pancreatic cancer cells. The small interfering RNA targeting SIRT1 and SIRT1 over-expression plasmid were transfected into the pancreatic cancer Panc-1 cells. Confocal microscopy was used to detect the LC3 expression. Western blot was used to analyze the protein levels of LC3, p62 and FOXO1/RAB7 signaling pathway-related molecules. Co-immunoprecipitation was used to detected the protein interaction between SIRT1 and FOXO1. RESULTS: The expression level of SIRT1 in the nucleus of Panc-1 cells was increased under hypoxia condition. Compared with negative control under hypoxia condition, knock-down of SIRT1 expression attenuated the autophagy flux in the pancreatic cancer Panc-1 cells (P<0.05). Over-expression of SIRT1 increased the protein levels of FOXO1 and RAB7. On the contrary, knock-down of SIRT1 expression inhibited the protein levels of FOXO1 and RAB7. The protein interaction between SIRT1 and FOXO1 in the pancreatic cancer cells was observed. CONCLUSION: SIRT1 in pancreatic cancer Panc-1 cells under hypoxia condition is over-expressed in the nucleus. Down-regulation of SIRT1 inhibits autophagy and its mechanism may be related to FOXO1/RAB7 signaling pathway.     

山地五月茶提取物对乳腺癌细胞MDA-MB-231增殖的抑制活性研究

为深入了解海南岛热带野生植物的保健功能及其活性成分,用CCK-8法检测山地五月茶（Antidesma montanum）果和叶的乙醇提取物对MDA-MB-231乳腺癌细胞增殖的抑制活性。结果表明,山地五月茶果和叶的乙醇提取物作用72 h对细胞的半数抑制浓度（IC₅₀）分别为483 μg/mL和306 μg/mL。叶的乙醇提取物分别经石油醚、乙酸乙酯、正丁醇萃取,比较各有机溶剂萃取相及剩余水相对乳腺癌细胞抑制增殖活性,结果显示乙酸乙酯相表现出较高的活性,其处理细胞72 h的IC₅₀为150.5 μg/mL。划痕实验和Transwell检测结果表明,乙酸乙酯相提取物具有抑制MDA-MB-231细胞迁移和侵袭的能力。流式细胞仪检测结果表明,乙酸乙酯相提取物能够使MDA-MB-231细胞周期S期显著延长。UPLC-MS/MS分析结果显示,乙酸乙酯相含有穗花杉双黄酮活性成分。穗花杉双黄酮处理MDA-MB-231细胞72 h的抑制增殖IC₅₀为192.6 μg/mL,表明其是山地五月茶抑癌活性成分之一。     

27-Hydroxycholesterol modulates lung cancer cell proliferation by activation of LXR signaling pathway

AIM:To investigate the effect of cholesterol metabolite 27-hydroxycholesterol (27-OHC) on the proliferation of lung cancer cells. METHODS:Human lung cancer A549 cells were treated with 27-OHC at different concentrations (0, 0.3125, 0.625, 1.25, 2.5, 5 and 10 μmol/L) for 24~48 h. The cell viability, cell cycle, cell prolife-ration, the intracellular cholesterol levels and cholesterol metabolism-related molecule expression were subsequently assessed by CCK-8 assay, flow cytometry, EdU staining, tissue total cholesterol detection kit, real-time PCR and Western blot. RESULTS:27-OHC decreased the viability of the A549 cells in a dose-and time-dependent manner (P<0.01) and inhibited the cell proliferation (P<0.05). The expression of typical liver X receptor (LXR) downstream target proteins including ATP-binding cassette transporter A1 (ABCA1), low-density lipoprotein receptor (LDLR), and 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-CR) were modulated, which promoted the efflux of intracellular cholesterol, and reduced cholesterol influx and de novo synthesis, resulting in decreased intracellular cholesterol levels and cell viability. Furthermore, the inhibitory effect of 27-OHC on A549 cell viability was significantly attenuated after the LXR pathway was partially blocked by 5 μmol/L GSK2033 treatment (P<0.05). CONCLUSION:27-OHC inhibits A549 cell prolife-ration via activation of LXR signaling pathway.     

Correlation between the histopathological diagnosis by AgNOR count and AgNOR area in canine mammary tumors

Background: Mammary tumors are the most common type of tumor in female dogs. The histopathological diagnosis is usually made by a hematoxylin-eosin (HE) staining of the tumor, which then requires a pathologist's judgment for assessment of malignancy. The purpose of this study was to investigate an alternative silver staining of some argyrophilic nucleolar organizer regions (AgNOR) for improving the diagnostic accuracy with mammary tumors.
Hypothesis: There is a correlation between the histopathological diagnosis by AgNOR count and AgNOR area in canine mammary tumors.
Animals: Seventy-three canine mammary tumors from 33 female dogs.
Materials and Methods: The AgNOR staining was evaluated retrospectively in 73 canine mammary tumors with a parallel HE staining as a "Gold Standard." Both a quantitative manual counting method and a qualitative computerized morphometric method were tested.
Result: The result from both methods indicated a clinically relevant difference in the mean values of the AgNOR in the following 4 categories: malignant, benign, hyperplastic, and normal mammary tissue. The counting method was superior, with 89% of the cases given a correct diagnosis of a malignant or a nonmalignant canine mammary tumor. The 2 methods were then compared to test their ability to classify the tumors correctly. Again, the counting method was the most reliable method, with a sensitivity of 80% and a specificity of 76% when the upper 50% of the AgNOR counts were presumed malignant.
Conclusion and Clinical Importance: The results indicated that an AgNOR test could be an aid to pathologists as a prognostic indicator or to assist them in deciding between a benign or a malignant diagnosis in questionable cases.     

云南松松塔提取物对H22实体瘤小鼠模型的抑瘤作用

研究云南松松塔乙醇提取物(PEA)和碱水提取醇沉物(PED)对H22实体瘤小鼠的抑瘤作用。建立H22实体瘤小鼠模型,随机分组,灌胃给药,1次/d,给药10d。末次给药24h后,取血检测γ干扰素(IFN-γ)、白细胞介素-2(IL-2)、白细胞介素-4(IL-4)和白细胞介素-10(IL-10)含量;剥离肿瘤组织并称重,计算抑瘤率;检测肿瘤组织匀浆中环氧化酶-2(COX-2)和前列腺素E2(PGE2)含量;石蜡包埋HE染色观察肿瘤细胞病理组织学变化。结果显示,PEA和PED各剂量组小鼠平均肿瘤重量较模型组显著减小;最大抑瘤率分别达46.11%和58.05%。PEA和PED各剂量组小鼠外周血中IFN-γ和IL-2含量较模型组显著升高;IL-4和IL-10含量较模型组显著降低。PEA和PED各剂量组小鼠肿瘤组织中COX-2和PGE2含量较模型组显著减少;给药组正常肿瘤组织减少,有出血和坏死,脂肪组织增多。结果表明,PEA和PED可抑制H22实体瘤小鼠的肿瘤生长,其机制与调节Th1/Th2细胞平衡和抑制COX-2与PGE2的表达有关。