ABSTRACT
Objective To explore the tumorigenic effect of interleukin(IL)-6 on mice with pancreatic carcinoma and the Caspase-3 / Bax / Bcl-2 signaling pathway related mechanism. Methods Forty mice were used to establish tumor-bearing animal model of pancreatic cancer with mouse pancreatic cancer cell line MPC-83. The tumor-bearing mice were divided into blank control group (A), intraperitoneal injection PBS 10 ml / kg; IL-6 group (B), intraperitoneal injection recombinant mouse IL-6 200 μg / kg; IL-6 receptor blocker group (C), intraperitoneal injection tocilizumab 100 mg / kg; and IL-6 + IL-6 receptor blocker group (D), intraperitoneal injection tocilizumab 100 mg / kg, 30 minutes later, intraperitoneal injection recombinant mouse IL-6 200 μg / kg, 10 mice in each group. The mice in each group were administrated corresponding drug once every 3 days for 28 days. The tumor volume was to observe and record at day 0, 7, 14, 21 and 28 after the experiment began. The mice were sacrificed by cervical vertebra dislocation after the last measurement of tumor volume. ELISA method was used to test the contents of survivin and cytochrome C (Cyt-C) in transplanted tumor tissue of mice. Reverse transcription polymerase chain reaction (RT-PCR) and Western blotting method were used to detect the expression levels of Caspase-3, Bax and Bcl-2 mRNA and protein in transplanted tumor tissue of mice. Results Compared with the A group, the transplanted tumor tissue of mice in group B grew rapidly on day 7, 14, 21 and 28, the content of survivin was increased, the content of Cyt-C was decreased, the expression levels of Caspase-3 and Bax mRNA and protein was down-regulated, and the expression levels of Bcl-2 mRNA and protein was up-regulated (P 0. 05). Conclusion The role of IL-6 in promoting the growth and proliferation of pancreatic cancer may be related to the regulation of Caspase-3/ Bax/ Bcl-2 cell apoptosis signaling pathway.
ABSTRACT
Objective:High performance liquid chromatography (HPLC) fingerprints of liposoluble and water-soluble fractions of Xiaojinwan were established and the similarity of fingerprints was evaluated, so as to explore the quality consistency of Xiaojinwan. Method:Chromatographic separation was carried out on Welch Ultimate AQ-C18 column (4.6 mm×250 mm, 5 μm) with the mobile phase of 0.1% phosphoric acid solution (A)-acetonitrile (B) for gradient elution (liposoluble fraction of 0-5 min, 40%B; 5-10 min, 40%-50%B; 10-20 min, 50%-60%B; 20-30 min, 60%-65%B; 30-40 min, 65%-70%B; 40-50 min, 70%-80%B; 50-60 min, 80%-90%B; 60-65 min, 90%-95%B; 65-75 min, 95%-100%B; 75-80 min, 100%B; water-soluble fraction of 0-20 min, 2%-5%B; 20-30 min, 5%-10%B; 30-37 min, 10%-20%B; 37-45 min, 20%-30%B; 45-50 min, 30%-40%B; 50-58 min, 40%B), the flow rate was 1 mL·min-1, the column temperature was 30 ℃. The detection wavelengths of the liposoluble and water-soluble fractions were 202, 250 nm, and their injection volumes were 10, 20 μL, respectively. A total of 30 batches of Xiaojinwan from five manufacturers were detected by HPLC, the chromatographic peaks of each part were analyzed by principal component analysis (PCA) and identified. Result:A total of 55 chromatographic peaks were detected in the fingerprints, and the similarity of fingerprint of 30 batches of Xiaojinwan was quite different. The relative standard deviations (RSDs) of fingerprint similarity of liposoluble and water-soluble fractions of Xiaojinwan were 21.5% and 32.8%, respectively. There were significant differences in the quality of samples from different manufacturers and the same manufacturer, and the chemical consistency evaluation results were dominated by liposoluble fraction, and the main reason for the chemical difference of this preparation was the composition of Liquidambaris Resina. Conclusion:The quality consistency of Xiaojinwan is poor. The establishment of two-fraction fingerprint provides a new idea for the overall quality evaluation and control of Xiaojinwan, and can provide a reference for the quality consistency evaluation of traditional pills.
ABSTRACT
OBJECTIVE@#Human U three protein 14a (hUTP14a) facilitates tumorigenesis through promoting p53 and Rb degradation as well as enhancing c-Myc oncogenic activity. Moreover, hUTP14a expression is up-regulated in human hepatocellular cancer and colorectal cancer tissues. In this study, the expression of hUTP14a in non-small cell lung cancer (NSCLC) tissues was evaluated by immunohistochemistry staining (IHC). The relationship between hUTP14a expression levels and the clinical characteristics of the NSCLC patients were analyzed.@*METHODS@#Lung cancer tissues and the adjacent non-cancerous tissues were collected from 123 cases of NSCLC patients including 53 cases of squamous cell carcinoma (SCC) and 70 cases of adenocarcinoma (ADC), who had accepted surgical resection at Peking University Third Hospital from May 2003 to April 2006. The expression level of hUTP14a was determined by IHC in human NSCLC tissues and the adjacent non-cancerous tissues. The associations between hUTP14a expression and the clinical pathological variables including gender, age, tumor size, histological type, differentiation degree and clinical pathological stage were analyzed using the Pearson's χ2 test.@*RESULTS@#The expression rate of hUTP14a in NSCLC tissues was significantly higher than that in the non-cancerous tissues (37.4% vs. 0, P<0.001). The expressions of hUTP14a in lung ADC and SCC were 48.6% and 20.6%, respectively. The expression rate of hUTP14a in both lung ADC and SCC was significantly higher than that in the adjacent non-cancerous tissues (P<0.001). In addition, the expression rate of hUTP14a in lung ADC was significantly higher than that in SCC (χ2=8.66, P=0.003). Furthermore, the expression rate of hUTP14a in the late pTNM stage of SCC was significantly higher than that in the early pTNM stage of SCC while hUTP14a expression level was not associated with pTNM stage of ADC. No correlation was found between hUTP14a expression and the other clinical pathologic features of the patients.@*CONCLUSION@#Expression of hUTP14a was up-regulated in NSCLC tissues and was correlated with pTNM stage of SCC, suggesting that hUTP14a might possess a potential as a candidate marker for the early diagnosis screening of NSCLC.
Subject(s)
Humans , Adenocarcinoma , Carcinoma, Non-Small-Cell Lung , Carcinoma, Squamous Cell , Lung Neoplasms , Prognosis , Ribonucleoproteins, Small Nucleolar/metabolismABSTRACT
Xiaojin pills, the first choice for clinical treatment of breast hyperplasia, were selected to explore the suitability of a bioactivity assay with chemical fingerprinting for the development of an overall quality evaluation assay. The liposoluble and water-soluble fraction fingerprints of Xiaojin pills were established. The ability to inhibit platelet aggregation and the rate of inhibition of cyclooxygenase-2 (COX-2) for 16 batches of Xiaojin pills from several manufacturers was analyzed; the chemical fingerprints of these samples were correlated with the bioactivity and chemical analysis. The animal protocol was approved by the Committee on the Ethics of Animal Experiments of Affiliated Hospital of Chengdu University of Traditional Chinese Medicine Approval, ID: 2018BL-002. Results showed that the antiplatelet aggregation activity of 16 batches was 0.712-1.278 U∙mg-1, with a relative standard deviation (RSD) of 15.4%. COX-2 inhibition was 52.07%-68.95% and the RSD was 8.91%. The results showed that there was little difference in the biological effects of these samples. However, the chemical fingerprint consistency of these 16 batches of Xiaojin pills was poor, and the similarity of nearly half of the samples was less than 0.9. The total peak area of Xiaojin pills was 32.74%-165.37% across samples, showing very poor chemical consistency. In order to explore the reasons for the poor chemical consistency despite good consistency in the biological assays, the fingerprint chromatogram was analyzed by multivariate statistical analysis. The main chromatographic peaks were identified. The results showed that the similarity of Xiaojin pills was mainly determined by the prominent chromatographic peaks 17, 18, 20, 23 and 27 in the liposoluble fingerprints, which were identified from Liquidambaris resina and Angelica sinensis Radix. However, Liquidambaris resina and Angelicae sinensis Radix had almost no anti-platelet aggregation activity or COX-2 inhibitory effect at the normal prescription ratio. As a result, the ability to utilize chemical fingerprints to evaluate the quality consistency of Xiaojin pills is limited. The selection of biological evaluation methods that reflect clinical efficacy could make up for the shortcomings of chemical evaluation methods for quality assessment, and provide new ideas and methods for the overall quality evaluation of complex Chinese patent medicines.
ABSTRACT
<p><b>OBJECTIVE</b>To investigate the regulatory effect of ATP?binding cassette transporter A1 (ABCA1) knockdown on inflammatory response induced by Pam3CSK4 in mouse mononuclear macrophage RAW264.7 cell line.</p><p><b>METHODS</b>A mouse mononuclear macrophage RAW264.7 cell line with stable ABCA1 knockdown was constructed and stimulated with Toll?like receptor 2 (TLR2) ligand Pam3CSK4, and the changes in the transcriptional levels of the proinflammatory and anti-inflammatory cytokines were analyzed in this cell model.</p><p><b>RESULTS</b>In RAW264.7 cells, ABCA1 knockdown significantly up-regulated Pam3CSK4 stimulation?induced expressions of IL?1β, TNF?α and IL?6 and also enhanced the expression of transcription factor cAMP?dependent transcription factor 3 (ATF3) without obviously affecting the expressions of the transcription factors ATF1, ATF2, ATF4 or ATF5.</p><p><b>CONCLUSION</b>ABCA1 knockdown in macrophages may have both proinflammatory and anti?inflammatory effects. ABCA1 knockdown up?regulates the transcription of ATF3 possibly through a mechanism that is different from that for the other members of the ATF protein family.</p>