ABSTRACT
ObjectiveTo analyze the transcriptome characteristics of Xianlian Jiedu prescription （XLJDP） in the intervention of colorectal carcinoma by high-throughput cDNA-sequencing （RNA-seq）. MethodNinety male C57BL/6 mice were randomly divided into the control group， colorectal carcinoma due to dampness， heat， stasis， and toxin model group， and XLJDP group， with 30 mice in each group. Mice in the model group and XLJDP group were fed a high-fat diet and provided with azoxymethane and dextran sodium sulfate （AOM/DSS） for inducing colorectal carcinoma. Those in the XLJDP group were further treated with intragastric administration of 12.9 g·kg-1 XLJDP since the day of modeling for 112 days. The colorectal tissues were collected from each group 4 h after the last drug treatment and stained with hematoxylin-eosin （HE） and methylene blue for observing the pathological changes. The total RNA was extracted from colorectal tissues for RNA-Seq-based transcriptome profiling， followed by gene oncology （GO） and Kyoto encyclopedia of genes and genomes （KEGG） pathway enrichment analysis and the screening and verification of differentially expressed genes. ResultCompared with the model group， XLJDP significantly relieved the colorectal congestion and edema and decreased tumor number and volume in mouse colorectal tissues. The methylene blue staining results indicated that XLJDP significantly suppressed the development of aberrant crypt foci （ACF，P<0.01）. As revealed by HE staining， XLJDP significantly alleviated the injury and dysplasia of colorectal tissues. Transcriptome analysis identified 615 differentially expressed genes （446 up-regulated and 169 down-regulated） between the model group and the blank group and 54 differentially expressed genes （29 up-regulated and 25 down-regulated） between the XLJDP group and model group. XLJDP mainly affected the expression of NIMA-related protein kinase 7 gene （Nek7， P<0.01）， Mucin 16 （Muc16， P<0.01）， SiahE3 ubiquitin protein ligase family member 3 （Siah3， P<0.01）， regenerating islet-derived protein 3-gamma （Reg3g， P<0.01）， RNA polymerase Ⅱ elongation factor-associated factor 2 （Eaf2， P<0.01）， transforming growth factor‐alfa gene （TGF-α， P<0.05）， secretoglobin family 1A member 1 （Scgb1a1， P<0.05）， family with sequence similarity 227 member B （Fam227B， P<0.05）， cytochrome P450 family 2 subfamily c polypeptide 40 （Cyp2c40， P<0.01）， and ankyrin repeat and EF-hand domain containing protein 1 （Ankef1， P<0.05）. Enrichment analysis showed that intestinal epithelial cell proliferation， metabolism of xenobiotics by cytochrome P450， and arachidonic acid metabolism signaling pathway were significantly enriched. ConclusionXLJDP is able to interfere with colorectal tumorigenesis and development due to dampness， heat， stasis， and toxin in mice， which has been proved by transcriptome analysis to be related to the regulation of metabolism-related pathways.
ABSTRACT
<p><b>OBJECTIVE</b>To evaluate the difference of 2 operation modes for patient plasma coagulation test and its intra-batch precision by using SysmexCA-7000.</p><p><b>METHODS</b>The SysmexCA-7000 blood coagulometer with agents, the normal and micro operation modes were respectively used to determine the prothrombin time (PT), fibringen (Fib), activated patial thromboplastine time (APTT) and thrombin time (TT) in 10 mixtures of multiple plasma samples, quality-control samples and 50 patient plasma samples, Among them, the 10 mixed samples and 2 quality-control plasma samples were tested for 10 times by every mode. The average level (̄X) of PT, Fib, APTT and TT, standard deviation (S) and intra-batch precision (coefficient of variation CV%) were respectively calculated. The intra-batch precision and coagulation results under 2 mode operations were analyzed statistically.</p><p><b>RESULTS</b>There was significantly statistical difference (P < 0.001) for intra-batch CVs between 2 mode for APTT, Fib, PT and TT with a level of 0.53%-1.58%, 0.72%-2.08%, 0.51%-1.37%, 0.58%-1.60% and Normal mode with 1.17%-2.10%, 1.10%-2.43%, 0.88%-1.99%, 1.05%-1.98%, respectively. APTT, PT and Fib of 50 patient plasma samples under micro mode operations were statistically different from normal mode, but TT was not different between 2 modes.</p><p><b>CONCLUSION</b>The micro mode detection is more accurate and precise than that of normal mode by using SysmexCA-7000 blood coagulometer.</p>
Subject(s)
Humans , Blood Coagulation Tests , Partial Thromboplastin Time , Plasma , Prothrombin Time , Thrombin TimeABSTRACT
The purpose of this study was to investigate the influence of blood coagulation reagents stored for different time on test results of the specimens prothrombin time (PT), activated partial thromboplastin time (APTT), thrombin time (TT) and fibrinogen (Fib). A total of 21 patient plasma specimens was taken and measured for homeostasis by Sysmex CA7000 automated blood coagulation analyzer and supporting reagent. The PT, APTT, TT and Fib of specimens were measured with the reagents stored in Sysmex CA7000 for different time. The differences of PT, APTT, TT and Fib were analyzed between values measured of the reagents stored for 0 hour and different time (TS:12, 24, 36,48, 60, 72 h; DA:24, 48, 72, 96, 120 h; TT:2, 4, 6, 8, 10, 12 h; TR:4, 8, 12, 16, 20, 24 h; OVB:1, 2, 3, 4, 5 ,6 h), respectively. The results showed that when coagulation reagent TS were stored for more than 48 h , DA 96 h, TT 10 h, TR 16 h and OVB 4 h, the values of PT, APTT, TT and Fib of samples were statistically different from the values measured with fresh coagulation reagent (P < 0.01), respectively. Compared 0 h with TS stored for 48-72, DA 96-120, TT 10-12, TR 16-24 and OVB 4-6 h, the percentage difference of PT, APTT, TT and Fib is in -2.6% ∼ 10.8%, -3.44% ∼ 4.8%, -3.9% ∼ 5.52%, -10.8% ∼ 3.3% and -17.2% ∼ 0.5%, the PT and Fib changes were more significant. Accordingly, the result of PT, APTT and TT had a uptrend as the reagent stored in Sysmex CA7000 analyzer for a long time, while Fib downtrend. It is concluded that the reagents showed be timely replaced when the plasma coagulation test is performed so as to obtain accurate results of examination.
Subject(s)
Humans , Blood Coagulation , Blood Coagulation Tests , Blood Preservation , Methods , Fibrinogen , Hemostasis , Indicators and ReagentsABSTRACT
The purpose of this study was to investigate the effect of glutathione (GSH) on blood coagulation. The normal plasma samples and mixed plasma samples were taken randomly, and into which the normal dose and different concentration of GSH were added. The prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen (FIB) and thrombin time (TT) were detected by using coagulation method before and after treatment with GSH. The detection results of normal plasma and mixed plasma containing GSH of different concentration were compared and analyzed with linear regression. The results showed that the APTT and FIB values of the plasma containing 2.5 mg/L glutathione or more, PT values of the plasma containing 10 mg/L glutathione or more, and TT values of the plasma containing 1250 mg/L glutathione or more were significantly different from those results of normal plasma or mixed plasma (P < 0.01) . There was a linear relation between all of the detection results of PT,APTT, FIB, TT and glutathione concentrations. The results of TT, APTT, PT and FIB detection in patient plasma were statistically different (P < 0.01) before and after treatment with normal concentration GSH. It is concluded that glutathione can influence detection results of coagulation function.