Pancreatic cancer is a highly lethal malignancy and one of the leading causes of cancer-related death. During the development and progression of cancer, tumor angiogenesis plays a crucial role. A great deal of evidence has revealed that human mast cells (MCs) contributed to tumor angiogenesis through releasing several pro-angiogenetic factors, among which tryptase is one of the most active. However, the role of mast cell tryptase (MCT) in human pancreatic cancer angiogenesis is still not well documented. In this study, we examined the MCT levels in serum from pancreatic cancer patients and evaluated the correlationship of the MCT level and tumor angiogenesis. In addition, the effect of MCT on endothelial cell proliferation and tube formation was investigated both in vitro and in nude mice bearing pancreatic tumor. It was found that MCT contributes to endothelial cell growth and tube formation via up-regulation of angiopoietin-1 expression. Moreover, using the MCT inhibitor nafamostat, tryptase-induced angiogenesis was obviously suppressed both in vitro and in vivo. Our findings suggest that MCT plays an important role in pancreatic cancer angiogenesis and tumor growth via activating the angiopoietin-1 pathway, and tryptase inhibitors may be evaluated as an effective anti-angiogenetic approach in pancreatic cancer therapy.
To estimate the age of skeletal muscle contusion, the expression of troponin I mRNA in contused skeletal muscle of rats was detected using real-time polymerase chain reaction (PCR). A total of 51 Sprague-Dawley male rats were divided into control and contusion groups, and another nine rats received contusion injury after death. At 0.5, 1, 6, 12, 18, 24, 30, and 36 h after contusion, the rats were killed with a lethal dose of pentobarbital. Total RNA was isolated from muscle specimens using the SV Total RNA Isolation System and reverse transcribed into first-strand cDNA. Sequence-specific primers were then used to conduct real-time PCR to analyze the expression levels of sTnI mRNA. At 0.5, 1, and 6 h after contusion, the expression levels of sTnI mRNA decreased to 78.17% (P < 0.05), 41.58% (P < 0.05), and 32.13% of that in the control group, respectively. However, there were no significant changes in the expression levels of sTnI mRNA from 6 to 36 h (P > 0.05) after contusion when normalized to RpL32 expression. The expression levels of sTnI mRNA in the normal and contused skeletal muscle of postmortem rats were about 70% of that in the control group (P < 0.05), and no significant changes in the expression levels of sTnI mRNA in the postmortem contusion group were noted among different time points after injury. This result suggests that determination of sTnI mRNA levels by real-time PCR is useful for the estimation of wound age.
Gene expression profiling by quantitative real-time PCR (RT-qPCR) is a valuable tool in forensic science for estimating the age of a wound. To accurately assess gene expression levels over time in injured tissue, the genes used as internal reference standards must be carefully validated for transcriptional stability. This study examined the transcriptional stability of nine potential reference genes (β-actin, GAPDH, RPL32, PGK1, SDHA, RPL13, HPRT, Tbp, and Ywhaz) in contused rat skeletal muscle by RT-qPCR. The raw Ct values were determined for each candidate gene at different time points following contusion, and the data were analyzed by the NormFinder, geNorm, and BestKeeper validation programs. The reference genes RPL13 and RPL32 were the most stably expressed genes in contused skeletal muscle, whereas PGK1 was the least stable. The commonly used reference genes β-actin and GAPDH appeared to be too unstable for normalization of RT-qPCR expression profiling in contused muscle. The reference genes RPL13 and RPL32 were also the best combination for multianalysis. The use of RPL13 and RPL32 as internal standards may improve the accuracy of gene expression studies aimed at determining the age of early wounds in forensic investigations.
To estimate the age of skeletal muscle contusion, the expression of SNAT2 mRNA in contused skeletal muscle of rats was detected by real-time polymerase chain reaction (PCR). In total, 78 Sprague-Dawley male rats were divided into control and contusion groups. At 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, and 48 h (n = 6) after contusion, the rats were sacrificed with a lethal dose of pentobarbital. Another 24 rats received contusion injuries at 6, 12, 18, and 24 h (n = 6) after death. Total RNA was isolated from muscle specimens using the TRIzol reagent and reverse-transcribed into first-strand cDNA. Sequence-specific primers and TaqMan fluorogenic probes for SNAT2 mRNA and RPL13 mRNA were designed using the AlleleID 6 software, and the expression levels of SNAT2 mRNA were determined by real-time PCR. At 4, 16, 20, and 24 h after contusion, expression levels of SNAT2 mRNA normalized to RPL13 mRNA increased by 2.07 (P < 0.05), 2.53 (P < 0.05), 2.68 (P < 0.05), and 2.06 fold (P < 0.05) respectively, versus that in the control group. However, there was no significant change in the expression level of SNAT2 mRNA from 24 to 48 h (P > 0.05) after contusion, when normalized to RPL13 mRNA. There was no change in the expression level of SNAT2 mRNA between the normal skeletal muscle from the left limb of the same injured rat and the control. Also, no degradation of SNAT2 mRNA was detected in the postmortem samples (P > 0.05). This result suggests that the determination of SNAT2 mRNA levels by real-time PCR may be useful for estimating wound age.
Background: Depression is a common mental disorder and the diagnosis is still based on the descriptions of symptoms. Biomarkers can reveal disease characteristics for diagnosis, prognosis, and treatment. In recent years, many biomarkers relevant to the mechanisms of depression have been identified. This study uses bibliometric methods and visualization tools to analyse the literature on depression biomarkers and its hot topics, and research frontiers to provide references for future research.Methods: Scientific publications related to depression biomarkers published between and were obtained from the Web of Science database. The BICOMB software was used to extract high-frequency keywords and to construct binary word-document and co-word matrices. gCLUTO was used for bicluster and visual analyses of high-frequency keywords. Further graphical visualizations were generated using R, CiteSpace and VOSviewer software.Results: A total of , articles related to depression biomarkers were identified. The United States ( . %) and China ( . %), which together account for more than half of all publications, can be considered the research base for the field. Among institutions,
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