Triptolide Inhibits Invasion and Tumorigenesis of Hepatocellular Carcinoma MHCC-97H Cells Through NF-κB Signaling.

BACKGROUND We investigated whether the plant-derived agent triptolide (TPL) could effectively inhibit the growth and invasion of human hepatocellular carcinoma (HCC) cells. MATERIAL AND METHODS MHCC-97H cells were treated with various concentration of TPL for various times. To detect the effect of NF-κB on TPL-induced signal pathways, MHCC-97H cells were transfected with p65 siRNA or p65 cDNA, then treated with TPL. We detected cell survival and apoptosis by MTT, soft-agar colony formation assay, flow cytometry, and TUNEL assay. Cell migration and invasion was determined by Matrigel invasion and a wound-healing assay. NF-κB activity was detected by electrophoretic mobility shift assay (EMSA); MMP-9 activity was detected by ELISA. Western blot and real-time PCR (RT-PCR) assays were used to detect p65 and MMP-9 protein and mRNA expression. A subcutaneously implanted tumor model of MHCC-97H cells in nude mice was used to assess the effects of TPL on tumorigenesis in vivo. RESULTS We showed that TPL treatment significantly suppressed growth and induced apoptosis of MHCC-97H cells in a dose- and time-dependent manner in vitro. Furthermore, TPL treatment inhibited invasion in vitro and inhibited the growth and lung metastasis of MHCC-97H cells in vivo. NF-κB and MMP-9 were inactivated with TPL treatment. Overexpression of p65 restored MMP-9 activity and inhibited the TPL anti-tumor effect on MHCC-97H cells. Knockdown of p65 blocked MMP-9 activation and enhanced TPL-induced cell apoptosis and survival inhibition, and TPL inhibition of migration and invasion in vitro. CONCLUSIONS TPL treatment inhibited MHCC-97H cell growth, invasion, and metastasis in vitro and vivo, suggesting that TPL could be developed as a potential therapeutic agent for the treatment of HCC.

Efficacy of trimetazidine and plasmin combined with alprostadil in treatment of lower extremity arteriosclerosis obliterans.

Clinical efficacy of trimetazidine and plasmin combined with alprostadil in the treatment of lower extremity arteriosclerosis obliterans was investigated. A retrospective analysis was performed on 132 patients with lower extremity arteriosclerosis obliterans treated in Yantai Yuhuangding Hospital from March 2015 to August 2017. Among them, 68 patients were treated with trimetazidine combined with alprostadil (group A), and 64 patients were treated with plasmin combined with alprostadil (group B). Patients were administered 2 courses of treatment and observed with regard to therapeutic effects, changes in blood flow perfusion indicators (vascular peak velocity and blood flow) of the superficial femoral artery, posterior tibial artery and dorsalis pedis artery, in endothelial function, in left ankle brachial index, in pain-free walking distance and in maximum walking distance. After treatment, the vascular peak velocity of group B patients was lower than that in group A (P<0.05), but the blood flow was higher than that in group A (P<0.05). After treatment, endothelial esterase, high-sensitivity C-reactive protein and circulating endothelial cell count levels after treatment were lower than those before treatment (P<0.05), but nitric oxide level was higher than that before treatment (P<0.05). After treatment, the left ankle brachial index was lower in group A of patients than that in group B (P<0.05). After treatment, the maximum walking distance was significantly higher in group A patients than that in group B (P<0.05). After treatment, the pain-free walking distance and maximum walking distance of the two groups of patients were higher than those before treatment (P<0.05). Both trimetazidine and plasmin combined with alprostadil can effectively treat lower extremity arteriosclerosis obliterans. The former is better than the latter in improving exercise capacity, but the latter is better than the former in improving blood flow perfusion in patients.

Network analysis of HBV­ and HCV­induced hepatocellular carcinoma based on Random Forest and Monte Carlo cross­validation.

Hepatocellular carcinoma (HCC) is one of the leading causes of cancer­associated mortality worldwide. Hepatitis B virus (HBV) and hepatitis C virus (HCV) are two common risk factors for HCC. The majority of patients with HCC present at an advanced stage and are refractory to therapy. It is important to identify a method for efficient diagnosis at early stage. In the present study gene expression profile data, generated from microarray data, were pretreated according to the annotation files. The genes were mapped to pathways of Ingenuity Pathways Analysis. Dysregulated pathways and dysregulated pathway pairs were identified and constructed into individual networks, and a main network was constructed from individual networks with several edges. Random Forest (RF) classification was introduced to calculate the area under the curve (AUC) value of this network. Subsequently, 50 runs of Monte Carlo cross­validation were used to screen the optimal main network. The results indicated that a total of 4,929 genes were identified in the pathways and gene expression profile. By combining dysregulated pathways with Z<0.05 and dysregulated pathway pairs with Z<0.2, individual networks were constructed. The optimal main network with the highest AUC value was identified. In the HCV group, the network was identified with an AUC value of 0.98, including 41 pairs of pathways, and in the HBV group, the network was identified with an AUC value of 0.94, including eight pairs of pathways. In addition, four pairs were identified in both groups. In conclusion, the optimal networks of HCV and HBV groups were identified with the highest AUC values. The use of these networks is expected to assist in diagnosing patients effectively at an early stage.

miR-221 promotes growth and invasion of hepatocellular carcinoma cells by constitutive activation of NFκB.

BACKGROUND AND OBJECTIVE: microRNAs (miRs) are small noncoding RNAs that modulate a variety of cellular processes by regulating multiple targets, which can promote or inhibit the development of malignant behaviors. Accumulating evidence suggests that microRNA-221 (miR-221) plays important roles in human carcinogenesis. It has recently found that miR-221 was overexpressed in hepatocellular carcinoma (HCC), and overexpression of miR-221 has a bad prognosis in these patients. Thus, down-regulation of miR-221 expression in HCC would provide new treatment approaches. This study aimed to study the role of miR-221 on HCC cell growth, apoptosis, invasion and metastasis in vitro and vivo, and explored the possible mechanisms involved. METHODS: Effects of miR-221 upregulation or miR-221 downregulation by miR-221 inhibitor (anti-miR-221) or miR-221 mimic (miR-221) transfection on growth, apoptosis and invasion of HepG2 cells in vitro was detected. Using p65 siRNA and p65 cDNA transfection to examine the NFκB signaling pathway. A subcutaneously implanted tumor model of HepG2 cells in nude mouse was used to assess the effects of anti-miR-221 or miR-221 overexpression on tumorigenesis development. Using an intravenously injected tumor model of HepG2 cells to assess the effects of anti-miR-221 or miR-221 overexpression on lung metastasis. The signaling pathway was analyzed in vivo. RESULTS: Anti-miR-221 inhibited growth, invasion and induced apoptosis of HepG2 cells in vitro. This was accompanied by concomitant attenuation of NFκB, and downregulation of NFκB downstream genes such as Bcl-2, VEGF and MMP-9. In addition, miR-221 overexpression promoted growth and invasion of HepG2 cells in vitro, and accompanied by activation of NFκB, and upregulation of NFκB downstream genes Bcl-2, VEGF and MMP-9. Targeting P65 or P65 overexpression reversed the effect of miR-221, and inhibited or induced miR-221 expression, creating a positive feedback loop in human HepG2, respectively. Morever, stable overexpression of anti-miR-221 in HepG2 cells inhibits establishment of xenografts and lung metastasis in nude mice; Stable overexpression of miR-221 in HepG2 cells promotes establishment of xenografts and lung metastasis in nude mice. CONCLUSIONS: Therapies targeting the miR-221 signaling pathway may be more effective to prevent primary tumor formation and organ metastasis. The ability of this therapy to decrease tumorigenesis and metastasis may be related to NFκB signals.

[Analysis of TNFbeta gene microsatellite polymorphism in Graves disease].

The work is to study the microsatellite polymorphism of tumor necrosis factor betagene(TNFc)in Graves disease. The TNFc microsatellite polymorphism in the first intron of tumor necrosis factor betagene was amplified by polymerase chain reaction. The allele frequency and genotype frequency of all samples were analyzed by polyacrylamide gel electrophoresis. The results showed that TNFc microsatellite polymorphism included two alleles and three kinds of genotype. The TNFc2 allele frequency in Graves disease group was greater than control group (chi2 = 4.02,P<0.05), indicating this allele may have an important role in the pathogenesis of Graves disease.

Extraskeletal myxoid chondrosarcoma: A report of two cases.

Extraskeletal myxoid chondrosarcoma (EMC) is a relatively rare but well-defined neoplasm. This report describes two patients, one with EMC of the buttock and one with EMC of the knee. The two cases presented with large lobed masses and long T1 and T2 signaling identified by magnetic resonance imaging (MRI). An enhanced MRI scan demonstrated enhancement of the tumors. The tumors were composed of strands or cords of oval and spindle cells embedded in abundant myxoid stroma. Pathology results confirmed EMC.