ABSTRACT
ObjectiveThis study aims to investigate the efficacy and underlying mechanism of Da Chaihutang （DCHT） in treating hepatocellular carcinoma （HCC） in vitro and in vivo. MethodWe employed methyl thiazolyl tetrazolium （MTT） assay and crystal violet staining to observe the proliferation of Hepa1-6 liver cancer cells treated with DCHT at different doses （0， 125， 250， 500， 1 000 mg·L-1） for different time periods （1， 2， 4， 8 days）. The orthotopic liver cancer model was established by injection of 1×106 Hepa1-6 cells into mouse， and then the model mice were randomly assigned into six groups： blank， model， DCHT （0.21， 0.625， 1.875 g·kg-1， ig， qd）， and positive control （5-fluorouracil， 25 mg·kg-1， ip， qod）. After 14 days of administration， the mice were sacrificed， and the liver samples were collected and fixed in 4% paraformaldehyde for hematoxylin-eosin （HE） staining. The Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP）， Cytoscape 3.7.2， STRING， and DAVID were used for the searching of the key targets of DCHT in treating HCC， the construction of protein-protein interaction （PPI） network， and the Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment. Quantitative real-time PCR was performed to determine the mRNA level of interleukin-6 （IL-6） in Hepa1-6 cells and liver tissue. Western blotting was employed to measure the protein levels of the proteins involved in the mitogen-activated protein kinase （MAPK） and signal transducers and activators of transcription 3 （STAT3） signaling pathways. ResultDCHT （500， 1 000 mg·L-1） treatment for 4 and 8 days inhibited the proliferation of Hepa1-6 cells in a dose- and time-dependent manner （P<0.05）. The in vivo assay showed that DCHT （high dose， 1.875 g·kg-1） treatment for 14 days led to high differentiation and unobvious heterogeneity of HCC cells and small necrotic area compared with the model group. Network pharmacology analysis predicted that the potential targets of DCHT in the treatment of HCC were mainly the inflammation cytokines such as IL-6， interleukin-1β （IL-1β）， and tumor necrosis factor-alpha （TNF-α） in HCC microenvironment. The potential signaling pathways involved in the treatment were mainly associated with HCC growth and differentiation， including MAPK and STAT3 signaling pathways. Compared with the blank group， DCHT （1 000 mg·L-1） treatment for 1， 2， 4， and 8 days down-regulated the mRNA level of IL-6 in Hepa1-6 cells （P<0.05）. Similar results were observed in the livers of mice treated with DCHT （0.625， 1.875 g·kg-1）. The in vitro assay demonstrated that DCHT （1 000 mg·L-1） treatment for 4 and 8 days and DCHT （500， 1 000 mg·L-1） treatment inhibited the phosphorylation of extracellular signal-regulated kinases 1/2 （ERK1/2）， c-Jun NH2-terminal kinase/stress-activated protein kinase （JNK）， p38 MAPK， and STAT3 in a dose- and time-dependent manner （P<0.05）. The in vivo assay showed that DCHT （0.625 and 1.875 g·kg-1） treatment only inhibited the phosphorylation of p38 MAPK and STAT3 （P<0.05）. ConclusionThe present study indicates that DCHT can inhibit liver cancer cell proliferation by regulating p38 MAPK/IL-6/STAT3 signaling pathway.
ABSTRACT
Objective To further improve the value of clinical application of a quantitative analysis method called LF-index (LFI) which based on real-time tissue elastography (RTE). Methods We prospectively enrolled 116 consecutive patients with chronic hepatitis B (CHB) and all patients underwent a liver biopsy and RTE between January 2015 and December 2015 at the First affiliated hospital of Wenzhou Medical University. Univariate and multivariate analyses were performed, and the prediction models for predicting significant fibrosis and cirrhosis were derived from independent predictors. Results (1) In multivariate analyses, spleno-portal index (SPI) (OR=13.956, P=0.002) and LFI (OR=6.283, P=0.023) were confirmed as independent predictors of significant fibrosis. In multivariate analyses of patients with and without cirrhosis, we found significant differences in the γ-Glutamyl transferase (GGT) (OR=1.012, P=0.049), SPI (OR=5.676, P=0.002) and LFI (OR=14.102, P=0.001). (2)A novel model called LFI-SPI score (LSPS) for prediction of significant fibrosis was developed (area under receiver operating characteristic curve [AUROC]=0.87), showing the superiority of diagnostic accuracy than LFI (AUROC=0.76, P=0.0109), aspartate aminotransferase to platelet ratio index (APRI) (AUROC=0.64, P=0.0031), fibrosis-4 index (FIB-4) (AUROC=0.67, P= 0.0044) and Fibroscan (AUROC=0.68, P=0.0021). (3) We also developed a LFI-SPI-GGT score (LSPGS) for predicting cirrhosis, with an AUROC of 0.93. The diagnostic accuracy of LSPGS was similar to that of Fibroscan (AUROC=0.85, P=0.134), and was superior to LFI (AUROC=0.81, P=0.0113), APRI (AUROC=0.67, P<0.0001), and FIB-4 (AUROC=0.72, P=0.0005). Conclusions We developed new formulas, LSPS and LSPGS for predicting significant fibrosis and cirrhosis in this prospective study. LSP score was mainly used for screening of significant liver fibrosis, and LSPG score was mainly used to exclude cirrhosis.
ABSTRACT
Blood viscosity was determined in 50 normal individuals. No significant difference was found between males and females, therefore the values were pooled.In 10 burn patients (burn extent ranged from 30% to 90%). blood viscosity was determined in the first 3 postburn days. All of them received whole blood as a part of the resuscitational regime. The amount of whole blood given was approximately TBSA% burnedxbody weight (kg)?0.3ml. The blood viscosity returned to within normal range in all cases on the 3rd postburn day.Blood viscosity was determined in 20 uninjured dogs to obtain its normal values. A 25% napalm burn was inflicted upon the animals, and they were divided into 2 groups of 7 each. Group A received the same fluid replacement protocol as what was given in human patients, while Group B animals were replenished with balanced salt solution in the amount of 5ml/1%TBS A/kg in the first 24 hours, and dextran was given as a colloid in the second 24 hours. No significant difference in blood viscosity was observed between these two groups during the observation period.The above findings suggested that whole blood as one of the constituents of replacement fluids for burn shock would neither enhance blood (?)scosity nor impede its recovery to normalcy.