To investigate the expression of human epidermal growth factor receptor 2 （Her2） and Split hairy enhancer 1 （Hes1） among chronic superficial gastritis （CSG）-intestinal metaplasia （IM）-intestinal type gastric cancer （IGC） in Correa cascade， exploring the molecular mechanism of IGC pathogenesis.
Methods
Immunohistochemistry （IHC） was used to detect the expression levels of Her2 and Hes1 proteins in 30 CSG， 41 IM， and 135 IGC specimens.Western blot was used to detect the expression levels of these two proteins in the gastric epithelial cells GES-1， human IGC cell lines NCl-N87， and MKN74. The spatial location information of Her2 and Hes1 proteins in CSG gastric gland was observed， their different expressions in the gastric pit，isthmus， neck， basal regions， IM， IGC， and different cell lines were compared One-way analysis of variance was used for comparison of means among multiple groups， and LSD t test was used for post hoc pairwise comparisons. Mann-Whitney U test was used to compare the medians of two groups， and Kruskal-Wallis （KW） H test was used to compare the medians of multiple groups with Bonferroni correction. Spearman rank analysis was used for correlation statistics.
Results
IHC results showed that the expression of Her2 （+++） in the isthmus， neck， basal region， IM and IGC was lower than that in the pit [1（3.3 %），0（0.0 %），0（0.0 %），4（9.8 %），10（7.4 %） vs 14（46.7 %）；all P ＜ 0.05］.The expressions of Her2 in the isthmus and IM was moderate， both higher than that in the neck， base，and IGC （P ＜ 0.001）. The expression of Her2 was the weakest in the neck， base， and IGC， and there was no significant difference among them （P ＞ 0.05）. The expression of Hes1 （++） in the isthmus，neck， basal region， IM and IGC was higher than that in the pit [9（30.0 %），4（13.3 %），21（51.2 %），39（28.9 %）比0（0.0 %）； all P ＜ 0.05］. The expression of Hes1 is strongest in the basal region，higher than that in the isthmus， neck， IM and IGC （P ＜ 0.001）. The expression of Hes1 in IM and IGC was stronger than that in the isthmus and neck， respectively （P ＜ 0.05）. There was no significant difference of the expression of Hes1 between IM and IGC， and between isthmus and neck （P ＞ 0.05）.The expression of Her2/Hes1 was significantly negatively correlated in the pit， isthmus， neck， basal region， IM， and IGC （rs = - 0.794， rs = - 0.300， rs = - 0.834，rs = - 0.921， rs = - 0.258， rs = - 0.528，all P ＜ 0.05）. Western blot results revealed that the expression of Her2/Hes1 was significantly negatively correlated in GES-1， NCl-N87 and MKN74 （rs = - 0.776， rs = - 0.671， rs = - 0.614， all P ＜ 0.05）.
Conclusions
In the Correa cascade from CSG to IM， eventually progressing to IGC，Her2 and Hes1 proteins exhibited a negative correlation. Compared with the pit， Her2 expression was gradually decreased with the progression of the disease， while Hes1 expression was moderately upregulated.
To investigate the anti-tumor effects of sarsasapogenin （SAR），a major constituent of Smilax china L.， on hepatocellular carcinoma and its potential molecular mechanism.
Methods
In this study， HepG2 cells were treated with SAR. Cell proliferation activity was assessed using the CCK-8 assay and EdU staining. Cell apoptosis was determined by flow cytometry. Cell migration and invasion were detected by transwell assay. Dfferential expressed gene functions， signaling pathways， and protein interactions were analyzed through RNA sequencing， GO，KEGG， and PPI analysis. The independent sample t test was used for comparison between two groups，the one-way analysis of variance was used for comparison between multiple groups， and the LSD-t test was used for pairwise comparison between mutiple groups.
Results
The CCK- 8 experiment revealed that SAR treatment significantly inhibited HepG2 cell proliferation with an increase of the concentration， and the IC50 value was 22.5 μg/mL. Compared to control （DMSO）， SAR inhibited cell proliferation， increased cell apoptosis level significantly， and led to a significant decrease in cell migration [（40.00 ± 2.00）% vs （80.00 ± 5.00）%］and invasion rates [（35.00 ± 2.00）% vs（70.00 ±4.00）%］ （all P ＜ 0.01）. RNA-seq showed significant changes in gene expression in HepG2 cells after SAR treatment. 1 308 genes were upregulated， while 2 966 genes were downregulated.The differentially expressed genes were significantly enriched in biological processes such as cell proliferation and signal receptor binding. Further KEGG analysis revealed that the differentially expressed genes mainly enriched in steroid biosynthesis， complement and coagulation cascade，TNF， MAPK， and NF-κB pathway. The hub genes CDK3， EPHA7， LRRK2， PRKCB， ANK3， TEK and ZNP70 were identified as the targets of SAR by PPI network analysis.
Conclusion
SAR could reduce the proliferation， migration and invasion and promote apoptosis of HepG2 cells. The underlying mechanism may involve the TNF signaling pathway， the NF-κB signaling pathway， and other related signaling pathways.
To explore the mechanism of the Forkhead box protein A2 （FoxA2）low-expressing hepatic progenitor cells response to differentiation reagent and their differentiation towards hepatocytes.
Methods
Rat liver progenitor cells were transfected with non-specific shRNA and FoxA2 shRNA and induced to differentiate towards hepatocytes by sodium butyrate or the combination of sodium butyrate and Ly294002， an inhibitor of PI3K/Akt signal pathway. Giemsa staining was used for morphology analysis， and glycogen staining was used for glycogen storage analysis. RNA sequencing was used to analyze differentially expressed genes between FoxA2 knockdown cells and the control cells， and KEGG was used to find the key signal pathway based on the differentially expressed genes. Western blot was used to detect the expression levels of FoxA2，albumin， Akt and phosphorylated Akt. RT-qPCR was used to detect the expression levels of albumin，HNF4α， Lama1， IL6， PI3K， and Myc mRNA. Independent-sample t-test was used to compared the difference between two groups； and ANOVA analysis was used to compare the differences among groups. LSD-t test was used for further comparison between two groups.
Results
Compared to control group， knocking down FoxA2 reduced the expression of albumin （0.27 ± 0.05 vs 1.01 ±0.30） and HNF4α mRNA （0.41 ± 0.10 vs 1.08 ± 0.30）. The glycogen synthesis function and albumin expression （0.59 ± 0.08 vs 1.09 ± 0.08） were reduced （P ＜ 0.01） in FoxA2 knockdown cells after sodium butyrate treatment when compared to those in the control group. RNA sequencing analysis found there were 1 243 upregulated expression genes in the FoxA2 knockdown cells， and KEGG analysis revealed that PI3K/Akt signal pathway is the pathway with the most upregulated expression genes. Compared with control group， the expression levels of Lama1 （5.49 ± 0.62 vs 1.01 ± 0.19）， IL6 （2.20 ± 0.10 vs 1.00 ± 0.06）， PI3K （1.44 ± 0.09 vs 1.02 ± 0.23） and Myc （1.44 ±0.19 vs 1.01 ± 0.14） and p-Akt （P ＜ 0.05） of Akt （0.95 ± 0.11 vs 0.71 ± 0.03） in FoxA2 knockdown cells were increased （P ＜ 0.05）. Furthermore， the combined treatment with Ly294002 and sodium butyrate increased the expression level of albumin （0.97 ± 0.10 vs 0.76 ± 0.03， P ＜ 0.05） and HNF4α mRNA （5.36 ± 0.34 vs 0.41 ± 0.15， P ＜ 0.01） in FoxA2 knockdown cells when compared with sodium butyrate-treated FoxA2 knockdown cells. In addition， glycogen synthesis function in FoxA2 knockdown cells treated with Ly294002 and sodium butyrate was better than that in sodium butyrate- treated FoxA2 knockdown cells， which was similar to that in the control group treated with Ly294002 and sodium butyrate.
Conclusion
Blocking PI3K/Akt signal pathway contributes to enhance the response to differentiation reagent of FoxA2 low-expressing liver progenitor cells to differentiate towards hepatocytes.
To investigate the effect of butorphanol（ BT） on neuroinflammation and Janus kinase 2（ JAK2）/signal transduction and activator of transcription 3（ STAT3） signaling pathwayin rats with cerebral ischemia reperfusion injury（ CIRI）.
Methods
A CIRI rat model was established by middle cerebral artery occlusion method. After modeling， the rats were randomly divided into sham operation group（ Sham group）， Model group， low dose BT group（ L- BT group，50 mg/kg）， medium dose BT group（ M-BT group， 100 mg/kg）， and high dose BT group（ H-BT group， 200 mg/kg）. Longa 5-point method was applied to assess the neurological deficits in rats；HE staining was applied to observe the pathological changes of brain tissue； the volume of cerebral infarction was measured by TTC staining； TUNEL staining was applied to detect the apoptosis rate； the levels of tumor necrosis factor-α （TNF-α）， interleukin （IL）-1β， IL-6， inducible nitric oxide synthase （iNOS）， IL-4， and IL-10 were measured by ELISA method； and Western blot was applied to detect the expression of inhibiting G protein （Gi）， stimulating G protein （Gs）， p-JAK2，JAK2， p-STAT3， and STAT3 proteins. The data were tested for normality and homogeneity of variance. If they met the requirements， one-way ANOVA was used for comparison among multiple groups， and LSD-t test was used for comparison between two groups. If not， the rank sum test was used for comparison between multiple groups， and the Bonferroni test was pairwise compared.
Results
Compared with Sham group， pathological damage of brain tissue in Model group was more serious， and the neurological function defect score， apoptosis rate， cerebral infarction volume， TNF-α[（352.47 ± 21.46） pg/mL vs （179.34 ± 10.21） pg/mL］， IL-1β[（42.27 ± 3.67） pg/ mL vs（20.31 ±1.54） pg/mL］， IL-6， iNOS， Gs， p-JAK2/JAK2 （0.82 ± 0.10 vs 0.25 ± 0.05）， p-STAT3/ STAT3（0.91 ± 0.06 vs 0.37 ± 0.06） were increased， while Gi， IL-4 and IL-10 were decreased （all P ＜0.05）. Compared with Model group， the pathological damage of brain tissue in L-BT， M-BT and H-BT groups was alleviated， the neurological function defect score， apoptosis rate， cerebral infarction volume， TNF-α [（294.53 ± 20.17） pg/mL， （257.29 ± 17.13） pg/mL， （210.14 ± 15.46） pg/ mL vs（352.47 ± 21.46） pg/mL］， IL-1β [（37.14 ± 2.11） pg/mL， （30.18 ± 2.04） pg/mL， （25.49 ± 1.73）pg/mL vs （42.27 ± 3.67） pg/mL］， IL-6， iNOS， Gs， p-JAK2/JAK2 （0.68 ± 0.07， 0.52 ± 0.06， 0.39 ±0.06 vs 0.82 ± 0.10）， p-STAT3/STAT3 （0.77 ± 0.05， 0.61 ± 0.04， 0.45 ± 0.03 vs 0.91 ± 0.06） were decreased， while Gi， IL-4 and IL-10 increased （all P ＜ 0.05）， and these changes were in a dose dependent manner.
Conclusion
BT can alleviate neuroinflammation and inhibit the JAK2/STAT3 signaling pathway in CIRI rats.
Islet transplantation is the most effective method to restore endogenous insulin secretion and control blood glucose， but there are still problems such as low survival rate， decline of long-term function and immune rejection of islet grafts. Mesenchymal stem cells （MSCs） have attracted much attention due to their ability to secrete the extracellular matrix （ECM） with a variety of cytokines， or promoting the restoration of cellular injury， angiogenesis and anti-inflammatory effects through direct cell-cell interaction. MSCs for islet transplantation has the advantages of wide source，low immunogenicity， and can effectively solve the problems of low survival rate， poor function and immune response of islet grafts， so they has become a research hotspot in islet transplantation. This paper reviews progress in MSCs in islet transplantation， including the role of MSCs and their secreted cytokines in islet culture in vitro， transplantation research in vivo and clinical application， as well as the problems in the application of MSCs， aiming to further promote the application of MSCs in clinical islet transplantation.
Wound healing is a complex and sophisticated biological process， the speed and quality of which could be affected by a variety of external and internal factors， such as infection，foreign bodies， necrosis and comorbidities. Among them， disorders in the body′s immune regulatory mechanism are the main factors that lead to wound healing. Therefore， effective regulation of the body′s immune system to control inflammation is essential for wound healing. Mesenchymal stem cells （MSCs） are a class of pluripotent stem cells originating from the mesoderm， which are widely used in the treatment of various clinical diseases due to their immunomodulatory and paracrine effects. In addition to their own proliferation and differentiation abilities， MSCs can also control the inflammatory environment of wounds and promote wound healing by regulating the proliferation，differentiation， and apoptosis of immune cells， including the regulation of macrophage polarisation and inhibition of neutrophil proliferation. In conclusion， this review provides an overview of MSCs′ability to promote wound healing by influencing the body′s immune cells and further regulating the body′s immune mechanisms.
Neuromuscular disorders （NMDs） are highly heterogeneous in etiology.Different genetic variants contribute to different disease entities. More than 800 genes for hereditary neuromuscular diseases have been reported. In addition， some disorders， such as congenital nemaline myopathy， is caused by variants of several different genes. Such genetic complexity of the neuromuscular disease presents significant challenges for in vitro disease modeling. However， due to the maintenance of the genetic background of the donor cell during cell reprogramming， induced pluripotent stem cells （iPSCs） has been used broadly in the studies of NMDs. In this paper， the progress in applications of iPSCs technology in the studies of NMDs are reviewed.
The translational research of mesenchymal stem cells （MSCs）in the treatment of respiratory system diseases has attracted widespread attention. They mainly exerts therapeutic effects through mitochondrial transfer， extracellular vesicles， and paracrine. In recent years， preclinical studies have found that MSCs migrate to lung tissues of respiratory system diseases and exhibit effective functions with repairing lung damage and inhibiting inflammation in the lung microenvironment.This review summarizes the recent progress of MSCs in the treatment of respiratory system diseases，expatiates their lung tissue distribution and therapeutic mechanism in respiratory system diseases models. This article will provide theoretical basis and strategies for the clinical research of MSCs in the treatment of respiratory system diseases.
