Plantamajoside modulates the proliferation, stemness, and apoptosis of lung carcinoma via restraining p38MAPK and AKT phosphorylation.

BACKGROUND: Plantamajoside (PMS), an active anti-inflammatory component and antioxidant derived from Herba Plantaginis, has been reported to exert a suppressive effect in liver cancer in vivo. In this study, we tested the effects of PMS on the metastatic 95D cell line. METHODS: 95D cells were characterized as most sensitive to PMS across several lung cancer cell lines. Cell viability within 24 h was tested with CCK-8. Different concentrations of PMS (0, 50, 100, and 200 µg/mL) and 5 µg/mL of cisplatin were established for later 24 h treatment. Relative mRNA and protein expression were assessed with PCR and Western blotting. Cell proliferation and stemness were indicated with colony and sphere formation. Cell metastasis was evaluated with wound healing and Transwell. Apoptotic cells and mitochondrial membrane potential were investigated with flow cytometry. RESULTS: CCK-8 assay showed PMS to inhibit the viability of 95D cells in a dose-dependent manner. PMS decreased colony formation and inhibited stemness in 95D cells. Invasion and migration were also inhibited. Moreover, PMS induced cell apoptosis, and decreased mitochondrial membrane potential. All of these effects were dose dependent. Interestingly, PMS treatment reduced the protein expression of p-p38 MAPK and p-AKT but not that of p38 MAPK and AKT. CONCLUSIONS: PMS inhibited proliferation, stemness, and migration, and initiated apoptosis in 95D cells, possibly through p38 MAPK and AKT dephosphorylation and mitochondria dysfunction. These findings support the promise of PMS as a prodrug in lung cancer treatment.

The overexpression of miRNA-212-5p inhibited the malignant proliferation of liver cancer cells HepG2 and the tumor formation in nude mice with transplanted tumor through down-regulating SOCS5.

BACKGROUND: This study aims to investigate the effect of miR-212-5p overexpression targeting suppressor of cytokine signaling 5 (SOCS5) on the malignant proliferation of liver cancer cells HepG2 and tumor formation in nude mice with transplanted tumors. METHODS: Luciferase reporter assay was used to detect the targeted relationship between miR-212-5p and SOCS5, and SOCS5 was overexpressed by the SOCS5 pcDNA vector. MiR-212-5p mimic and pc DNA-SOCS5 were transfected into liver cancer HepG2 cells alone or in combination, and the cells were randomly divided into four groups, the control group, mimic group, SOCS5 group and mimic + SOCS5 group for subsequent experiments. The orthotopic xenograft mouse models were established by using HepG2 cells in BALB/c athymic nude mice. RESULTS: The results showed that there was a direct targeting relationship between miR-212-5p and SOCS5. Compared with the control group, the clone formation rate, the levels of Ki67, and proliferating cell nuclear antigen (PCNA) protein in the mimic group were significantly lower (P<0.05), but the apoptosis rate was significantly higher (P<0.05). The ratio of Bax/Bcl-2, cleaved Caspase-3/Caspase-3, and cleaved Caspase-9/Caspase-9 was significantly higher (P<0.05), while the ratios of p-phosphatidylinositol 3 kinase (PI3K)/PI3K, p- Protein kinase B (AKT)/AKT, and p-mammalian target of rapamycin (mTOR)/mTOR were significantly reduced (P<0.05). In the SOCS5 group, the result was reversed. Interesting, In the mimic+SOCS5 group the clone formation rate, the protein levels of Ki67, and PCNA were significantly decreased (P<0.05) while the apoptosis rate was significantly increased (P<0.05). The ratio of Bax/Bcl-2, cleaved Caspase-3/Caspase-3, and cleaved Caspase-9/Caspase-9 was significantly increased (P<0.05). The ratios of p-PI3K/PI3K, p-Akt/AKT, and p-mTOR/mTOR were significantly reduced (P<0.05). In vivo, The level of miR-212-5p was significantly increased, with SOCS5 decreased (P<0.05). Furthermore, the number of Ki67 positive cells was significantly reduced (P<0.05), and the apoptosis rate increased significantly (P<0.05). Additionally, the ratio of p-PI3K/PI3K, P-AKT/AKT, P-mTOR/mTOR decreased significantly (P<0.05). CONCLUSIONS: miR-212-5p overexpression down-regulated SOCS5 could inhibit the malignant proliferation of HCC cells HepG2 and tumor formation in nude mice with transplanted tumors.

Role of prophylactic filter placement in the endovascular treatment of symptomatic thrombosis in the central veins.

OBJECTIVES: To evaluate the role of filter implantation in reducing the incidence of fatal pulmonary embolism during the endovascular treatment of thrombosis in the major tributary of the superior vena cava (SVC). METHODS: From October 2004 to October 2008, we conducted a cohort study of 40 patients with thrombosis of the central veins who were preparing for endovascular interventions and received or did not receive filter. The symptom scores were measured, the incidence of pulmonary embolism (PE) was observed, and patient follow-up studies were conducted for three years. RESULTS: One week after therapy, the symptom score improved in both groups compared with before therapy (P <0.001), but no significant difference was found between the scores of the two groups (P >0.05). Four patients in the control group died from PEs after therapy, but no patients in the filter group presented evidence of PE. The survival rates at 1, 2, and 3 years (72.9%, 50%, and 27.1%, respectively) for the filter group were higher than those for the control group (47.6%, 19.0% and 14.3%, respectively; P =0.015). The survival time of patients in the filter group with bronchogenic carcinoma (18 ± 2 months) was longer than that of the patients in the control group (12 ± 2 months) after the endovascular treatment (P <0.001). CONCLUSIONS: Prophylactic filter placement could be a safe and effective method for preventing PE in pre- or post-endovascular-treated patients with thrombi in their central veins.

[Analysis of three metabolite residues of dipyrone in bovine muscle and pork muscle using high performance liquid chromatography].

A method for the determination of metabolite residues of dipyrone, 4-formylaminoantipyrine (FAA), 4-aminoantipyrine (AA) and 4-methylaminoantipyrine (MAA) in bovine muscle and pork muscle has been developed. Homogenized muscle sample was extracted with Na2SO4-Na2SO3 solution. After filtration, the extract was cleaned-up by a C18 solid phase extraction cartridge. Prepared sample was determined using high performance liquid chromatography under the following conditions: Inertsil ODS-3 column, methanol and water as mobile phase with gradient elution, detection wavelength of 265 nm. FAA was quantified by external standard method, while AA and MAA were quantified by internal standard method using isopropylaminoantipyrine as internal standard. Limits of detection of FAA was 12.5 microg/L, while those of AA and MAA were 15.0 microg/L and 20.0 microg/L, respectively. The limit of detection of this method was 50 microg/L for all the three metabolites. The recoveries of FAA, AA and MAA were 81.3% - 92.5%, 82.0% - 96.0%, and 80.4% - 90.6%, respectively, at the spiked levels of 50 - 400 microg/kg. The relative standard deviations of the method were less than 7%.

[Determination of 25 organochlorine pesticides in tea by gas chromatography-mass spectrometry].

A high performance gas chromatography-mass spectrometry (GC-MS) method for the determination of 25 organochlorine pesticides in tea has been developed. The samples of tea were extracted with n-hexane-acetone (2: 1, v/v). The extract was purified by using a Florisil column with n-hexane-ethyl acetate (9: 1, v/v) as elution solvent. Chromatographic analysis was performed on a DB-35MS capillary column. Satisfactory separation and sensitivity of 25 organochlorine pesticides were obtained with the proposed method. The analytical results show the working curves for 25 organochlorine pesticides were linear in the range of 0.010 - 0.500 mg/L by GC-MS on selective ion monitoring mode. The recoveries of 25 organochlorine pesticides at spiked levels of 0.01 -0.20 mg/kg were 70.8% - 105.5%, and relative standard deviations (RSDs) were 1.6% - 12.7%. The limits of quantitation were 0.01 mg/kg except that for endosulfan I and endosulfan II that were 0.02 mg/kg.

[Effects of intermittent and continuous thermochemotherapeutic intra artery infusion on adriamycin concentration in rabbit VX-2 tumor].

BACKGROUND & OBJECTIVE: It has been proved that vital signs of organism can be influenced by heat infusion and the thermochemotherapy with Adriamycin (ADM) is more effective than the general chemotherapy in inhibiting extraneous rabbit VX-2 cells. Intermittent thermochemotherapeutic infusion and continuous thermochemotherapeutic infusion with ADM were performed respectively on the rabbits to evaluate the effectiveness and safety of intermittent thermochemotherapeutic intra artery infusion by comparing their respiration rate, heart rate, body temperature, and the ADM concentration in VX-2 carcinoma. METHODS: VX-2 tumor models were established in the hind legs of 30 New Zealand rabbits, and then they were divided into three groups (10 in each group) randomly. 100 ml saline and ADM in room temperature were infused, 100 ml saline and ADM in 60 degrees C were intermittently infused, and 100 ml saline and ADM in 60 degrees C were continuously infused into the tumor nutrient arteries, which were confirmed by DSA, of the rabbits in each group respectively. During the infusion, the 43-45 degrees C lasting time of the tumor tissues in the two 60 degrees C infusion groups was measured. After the infusion,the respiratory rate,heart rate,body temperature,and the concentration of ADM within the tumors were determined. RESULTS: The concentration of ADM was 7.115+/-2.180 microg/ml in the room temperature infusion group,17.213+/-1.657 microg/ml in the 60 degrees C continuous infusion group, and 16.545+/-3.426 microg/ml in the 60 degrees C intermittent infusion group. There was no significant difference between the 60 degrees C intermittent infusion group and the 60 degrees C continuous infusion group (P >0.05), while there was significant difference between the 60 degrees C intermittent infusion groups and the room temperature infusion group,so was between 60 degrees C continuous infusion groups and the room temperature group (P< 0.05). The 43-45 degrees C lasting time was 22.53+/-1.44 minutes in the continuous infusion group and 24.31+/-2.45 minutes in the intermittent infusion group. There was no significant difference between these two groups (P >0.05). There was no significant difference in the respiration rate, heart rate, and body temperature between the 60 degrees C intermittent infusion group and the room temperature infusion group (P >0.05). CONCLUSION: Compared with continuous infusion, intermittent thermochemotherapy intra artery infusion is a more effective and safer interventional thermochemotherapy.

[Adriamycin thermotherapy through hepatic artery for model of VX2 carcinoma in rabbit liver].

BACKGROUND & OBJECTIVE: It was reported that heating can enhance sensitivity of rabbit VX2 cell to adriamycin and increase intracellular concentration of adriamycin. This study was designed to evaluate the anti-tumor effects of interventional hyperthermia and interventional chemotheramotherapy on VX2 carcinoma in rabbit liver. METHODS: VX2 carcinoma cells were surgically implanted into the right liver lobe of 60 male New Zealand white rabbits, which were randomly divided into 4 groups(15 rabbits per group). To inject physiological saline(37 degrees C), adriamycin (37 degrees C), physiological saline(60 degrees C), and adriamycin (60 degrees C) in different groups via hepatic artery of the rabbits with liver cancer. One week later, to observe the volume of tumor, the serum level of aspartate transaminase(AST), and observe the survival period of VX2 rabbits. RESULTS: In group of ADM(60 degrees C), the tumor growth rate (0.53 +/- 0.21)% was significantly lower than group 2(1.09 +/- 0.26)%, group 3(3.32 +/- 1.28)%, and group 4(3.48 +/- 1.17)% (P < 0.05, P < 0.05, P < 0.01, respectively). The survival period of adriamycin (60 degrees C) group (50.0 +/- 2.0)d was significantly higher than the untreated control group (40.5 +/- 3.0)d, (P < 0.05). The serum level of AST of TNP-470 with lipiodol group was not higher than the other treated groups(P > 0.05), but being significantly higher than the untreated control group after treated(P < 0.05). CONCLUSION: Adriamycin (60 degrees C) greatly decreases the tumour growth rate, and prolongs the survival period.