AIM:To investigate the effect of suberoylanilide hydroxamic acid ( SAHA) on the proliferation and apoptosis of human hepatocellular carcinoma HepG 2 cells and to explore its possible mechanism .METHODS: HepG2 cells were treated with SAHA at different concentrations for 48 h.The proliferation of HepG2 cells was detected by real-time cellular analysis.The protein levels of acetylated histones H3K9 and H3K27, glucose-regulated protein 78 (GRP78), protein kinase R-like endoplasmic reticulum kinase ( PERK ) and p-PERK were determined by Western blot .The cell apoptosis was analyzed by flow cytometry .RESULTS:Compared with control group , treatment with SAHA at 0.1μmol/L and 1 μmol/L for 48 h showed no significant inhibitory effect on the proliferation of HepG 2 cells, while SAHA at 6 μmol/L and 12 μmol/L significantly inhibited the proliferation of HepG 2 cells (P<0.05).The results of Western blot showed that the protein levels of acH3K9, acH3K27, GRP78 and p-PERK increased significantly after treated with SAHA at diffe-rent concentrations for 48 h, while the protein level of PERK was decreased significantly (P<0.05).The results of flow cytometry analysis showed that the apoptotic rates of the HepG 2 cells increased with the increase in SAHA concentration . CONCLUSION:SAHA up-regulates the acetylation of H3K9 and H3K27 in the HepG2 cells and induces apoptosis of HepG2 cells by activating the endoplasmic reticulum stress-related apoptosis pathway .

AIM:To investigate the role of microRNA-101-3p (miRNA-101-3p) on the proliferation, apopto-sis and invasion of gastric cancer cells and the possible regulatory mechanisms .METHODS: The expression of miRNA-101-3p in two kinds of gastric cancer cells and a gastric mucosal cell line was detected by real -time PCR.The miRNA-101-3p was overexpressed by Lipofectamine 2000 transfection with miRNA-101-3p mimics.The effects of miRNA-101-3p on cell cycle distribution and apoptosis were analyzed by flow cytometry .The effects of miRNA-101-3p on cell proliferation and migration abilities were detected by CCK-8 assay, trypan blue exclusion test and Transwell assay .The protein expression of enhancer of zeste homolog 2 ( EZH2) was determined by Western blot .RESULTS: The expression of miRNA-101-3p in gastric cancer cells was lower than that in gastric mucosal cells (P<0.05).The gastric cancer cell MGC-803 had the low-est expression level of miRNA-101-3p.The result of flow cytometry showed that the population of S phase was reduced , and the population of G0/G1 phase and the early stage apoptotic rate were increased after the expression of miRNA-101-3p was overexpressed (P<0.05).The results of CCK-8 assay, trypan blue exclusion test and Transwell assay showed that overex-pression of miRNA-101-3p significantly reduced the proliferation and migration abilities of gastric cancer cells (P<0.05). Overexpression of miRNA-101-3p decreased the protein level of EZH2 (P<0.05).CONCLUSION:miRNA-101-3p may suppresses the gastric cancer cell proliferation and migration , and promotes the gastric cancer cell apotosis by down-regula-tion of EZH2.

AIM:To investigate the effect of signal transducer and activator of transcription 3 (STAT3) on air-way injury in asthma and its mechanism .METHODS:The expression and activation of STAT 3 in bronchial mucosa tissues of asthmatic patients were measured by Western blot .The expression and activation of STAT 3 in bronchial epithelial cells pretreated with Dermatophagoides pteronyssinus antigen P1 (Derp1) were estimated.Bronchial epithelial cells were trans-fected with STAT3 shRNA.STAT3 expression was measured by Western blot .The cell viability was detected by CCK-8 as-say.The concentrations of TNF-α, IL-1βand IL-6 were detected by ELISA .The content of malondialdehyde ( MDA) and the activity of superoxide dismutase ( SOD) and glutathione peroxidase ( GSH-Px) were also determined for evaluating the status of oxidative stress .The cell apoptosis was analyzed by flow cytometry .RESULTS:The protein level of p-STAT3 was significantly up-regulated in both bronchial mucosa of asthmatic tissues and bronchial epithelial cells pretreated with Derp 1. Knockdown of STAT3 inhibited the releases of TNF-α, IL-1βand IL-6, decreased the content of MDA and enhanced the activity of SOD and GSH-Px with the suppression of cell apoptosis ( P <0.05 ) .CONCLUSION: Down-regulation of STAT3 attenuates Derp1-induced the airway injury .The mechanism may involve that knockdown of STAT3 suppresses the activation of JAK/STAT signaling pathway , the release of asthma-related inflammatory cytokines and oxidative stress in bronchial epithelial cells , thus inhibiting cell apoptosis .

AIM: To investigate whether Toll-like receptor 4 ( TLR4 ) and Nod-like receptor protein 3 (NLRP3) inflammasome were involved in contrast medium (CM)-induced inflammation and injury in renal tubular epithe-lial cells.METHODS: Iopromide was used to injure NRK-52E cells in the study.The cell viability was measured by CCK-8 assay.The protein levels of TLR4, NLRP3, apoptosis-associated speckle-like protein (ASC), caspase-1 and cleaved caspase-3 were determined by Western blot .The releases of interleukin ( IL )-1βand IL-18 were detected by ELISA .The apoptotic rate was evaluated by Hoechst staining , and mitochondrial membrane potential ( MMP) was analyzed by JC-1 staining.siRNA was transfected into the NRK-52E cells to silence NLRP3 expression.RESULTS:CM decreased the viability of NRK-52E cells (P<0.05).CM also elevated the protein levels of cleaved caspase-3, TLR4, NLRP3, IL-1βand IL-18 (P<0.05).Silencing NLRP3 attenuated CM-induced releases of inflammatory cytokines .Moreover, treat-ment with TLR4 inhibitor TAK-242 or knockdown of NLRP3 by siRNA transfection both attenuated cell apoptosis and loss of MMP caused by CM .CONCLUSION:TLR4/NLRP3 inflammasome takes part in the pathogenesis of CM-induced acute kidney injury , and mediates CM-induced injury and inflammation in renal tubular epithelial cells .

AIM: To observe the cardiac function during high-altitude exposure in Chongqing soldiers and to discuss its relationship with acute mountain sickness ( AMS ) by echocardiography .METHODS: The changes of heart function were evaluated during acute high-altitude exposure (3658 m, 3 d) in 42 healthy young male soldiers by echocar-diography.At the same time, the heart rate, blood pressure, blood oxygen saturation, and the incidence of AMS after high-altitude exposure were observed and recorded .RESULTS:Three days after arrival at 3658 m, the left atrial end-systolic dimension (LADs), left ventricular end-diastolic dimension (LVDd) and right atrial end-systolic dimension were signifi-cantly decreased , but the right ventricular outflow tract diameter , pulmonary artery dimension , ejection fraction , cardiac output (CO), pulmonary arterial systolic pressure (PASP) and mean pulmonary arterial pressure were significantly in-creased compared with the baseline levels in all subjects .The mitral peak E velocity was significantly reduced (P<0.05). A total of 42 healthy young men were recruited and divided into AMS group with 15 subjects and non-AMS group with 27 subjects by Lake Louise scoring after high-altitude exposure .The cardiac function in the plain showed that aortic sinus di-ameter and LVDd in AMS group were significantly smaller , and PASP was significantly higher than those in non-AMS group.After high-altitude exposure, the LADs in AMS group was significantly smaller than that in non-AMS group (P<0.05).AMS scores and CO in the plain showed significant negative correlation (r=-0.3814, P<0.05).CONCLU-SION:Upon acute high-altitude exposure , right ventricular functions of the young male soldiers are damaged with the com-pensation of the left ventricular functions .Using echocardiography to observe PASP and CO may be helpful for screening the susceptible people of AMS in the plain .

AIM:To investigate the effect of Eph receptor A2 (EphA2) on drug resistance of colorectal carci-noma cells and its possible mechanisms .METHODS:Real-time PCR and Western blot were used to detect the expression of EphA2 at mRNA and protein levels in LoVo and LoVo/5-FU cells.EphA2 siRNA was transfected to down-regulate the EphA2 expression in LoVo/5-FU cells, and the drug sensitivity was calculated by CCK-8 assay.Meanwhile, cell migration and invasion were measured by wound healing assay and Transwell assay , and the protein levels of E-cadherin,β-catenin, N-cadherin, vimentin, Notch and Snail were determined by Western blot .RESULTS: The expression of EphA2 at both mRNA and protein levels was significantly up-regulated in LoVo/5-FU cells (P<0.05).Knockdown of EphA2 suppressed the cell viability, and migration and invasion abilities , but promoted drug sensitivity of LoVo/5-FU cells.Up-regulation of E-cadherin and β-catenin, and down-regulation of N-cadherin and vimentin were observed , indicating that the epithelial-mesenchymal transition ( EMT) process was suppressed .Knockdown of EphA2 decreased the expression levels of Notch and Snail.CONCLUSION:Down-regulation of EphA2 partly reverses drug resistance of LoVo/5-FU cells.The mechanism may be related to suppressing cell growth , migration, invasion and EMT process via Notch/Snail signaling pathway .

AIM: To synthesis and characterize a multi-functional siRNA delivery agent with effective thera-peutic effects and MR-tracing ability for programmed death ligand-1 ( PD-L1 ) positive gastric cancer SGC-7901 cell line . METHODS:The characterization , binding ability , cytotoxicity , transfection efficiency and cellular internalization of the polyplex were determined .The PD-L1 knockdown effect was analyzed , and cytokines secreted by cocultured T cells were measured.RESULTS:We developed folic acid (FA)-PEG-SS-PEI-SPION as siRNA delivery agent for PD-L1 knockdown. At N/P ratio of 10, the FA-PEG-SS-PEI-SPION bound PD-L1 siRNA to form polyplex in a diameter of (116.7 ±2.5) nm with zeta potential of (9.14 ±0.80) mV.Transfection efficiency of the targeted polyplex was (95.06 ±0.44)%, com-pared with ( 93.87 ±1.05 )% of the untargeted polyplex .Mean fluorescence intensity of the targeted polyplex was 1892.67 ±81.51, significantly higher than 1324.33 ±186.58 of the untargeted.The cellular magnetic resonance (MR) imaging showed the polyplex also acted as T 2 weighted contrast agent for cancer MR imaging .The relative mRNA level of PD-L1 in polymer/siRNA-2 treatment group was (9.07 ±0.79)%.Decreased protein expression of PD-L1 was showed by Western blot .The secretion levels of IFN-γand TNF-αin cocultured T cells increased , while that of IL-10 decreased . CONCLUSION:Our findings highlighted the potential of the multifunctional theranostic nanoparticles for effective targe -ting PD-L1 knockdown therapy and MR imaging diagnosis in gastric cancers .

AIM:To investigate the effects of exosomes secreted by pancreatic cancer cells on the viability and function of βcells and the possible mechanism .METHODS:ExoQuick-TC kit was used to extract exosomes in the super-natants of mouse pancreatic cancer Pan 02 and MPC-83 cells, and the extracted exosomes were identified by transmission electron microscopy.Fluorescence-labeled exosomes were incubated with mouse insulinoma MIN 6 cells for 48 h to detect whether exosomes secreted by pancreatic cancer cells were uptaken by MIN 6 cells.MTT and glucose-stimulated insulin se-cretion ( GSIS) assays were conducted to examine cell viability and insulin secretion of MIN 6 cells after incubating with ex-osomes.The expression of miR-204 and Bcl-2 mRNA in MIN6 cells was detected by qPCR .The protein expression of Bcl-2, Bax, caspase-3 and cytochrome C (Cyt-C) in MIN6 cells was determined by Western blot .RESULTS:The results of transmission electron microscopy showed that both Pan 02 cells and MPC-83 cells secreted exosomes , and Pan02 cells secre-ted more.The co-incubation results of fluorescence-labeled exosomes and MIN6 cells confirmed that MIN6 cells were able to ingest large amounts of exosomes secreted by pancreatic cancer cells .The results of MTT and GSIS assays showed that the viability and the level of high glucose-stimulated insulin secretion of MIN 6 cells in exosome treatment group significantly decreased compared with nontreatment group (P<0.01).The results of qPCR showed that the exosomes secreted by pan-creatic cancer cells were rich in miR-204, and the mRNA expression of Bcl-2 in MIN6 cells was significantly down-regula-ted by exosome incubation ( P<0.01) .The results of Western blot showed that the protein expression of Bcl-2 in the MIN6 cells treated with exosomes was significantly down-regulated (P<0.05), and the protein levels of Bax, cleaved caspase-3 and Cyt-C in exosomes treatment group were significantly up-regulated ( P<0.01 ) .CONCLUSION: Pancreatic cancer cells secrete exosomes .The exosomes secreted by pancreatic cancer cells are ingested by βcells, and reduce the viability and insulin secretion of βcells.The mechanism may be related to the increase in exosomal miR-204 in the βcells.In-creasing miR-204 may inhibit the expression of Bcl-2 and promote the activation of mitochondrial apoptosis in βcells.

AIM:To observe the effects of exogenous zinc on the biological behavior of hepatocellular carcino -ma ( HCC) cell line BEL-7404.METHODS: BEL-7404 cells were cultured with zinc sulfate at various concentrations . The intracellular concentration of zinc , cell viability , cell cycle , cell apoptosis and migration and invasion abilities were measured by TSQ fluorescent probe , MTT assay, DNA ploid analysis, acridine orange/ethidium bromide fluorescence stai-ning and Transwell assay , respectively .The mRNA and protein expression levels of albumin in the BEL-7404 cells were determined by real-time PCR and Western blot , respectively .RESULTS: With the elevated concentration of zinc in cul-ture condition, the concentration of zinc in the BEL-7404 cells was increased (P<0.05).The cell viability and migration and invasion abilities were decreased , while the apoptotic rate was increased (P<0.05).The cells in G0/G1 phase were decreased, while the cells in G2/M phase were increased.Additionally, the mRNA and protein expression of albumin also increased (P<0.05).CONCLUSION:The zinc ion inhibits the cell viability as well as migration and invasion abilities , blocks the cells in G 2/M phase , and may reduce cell malignant phenotype .

AIM:To identify the expression of fermitin family homolog 2 (FERMT2) in hepatocellular carci-noma ( HCC) tissues and the effect of FERMT 2 on the cell growth and related protein expression .METHODS:Real-time PCR and immunohistochemistry were used to detect FERMT 2 expression in the HCC tissues .The technique of CRISPR/Cas9 was applied to construct stable FERMT2 knockout MHCC97H cell line.WST-1 assay and flow cytometry were used to measure the cell viability , cell-cycle distribution and cell apoptosis .Western blot was used to determine the expression of related proteins in the MHCC97H cells.RESULTS:In HCC tissues, the expression level of FERMT2 was higher than that in adjacent liver tissues (P<0.05).High expression of FERMT2 was significantly correlated with postoperative recurrence of tumor.Knockout of FERMT2 gene evidently inhibited MHCC97H cell viability and accelerated cell apoptosis .Mean-while, the expression levels of proliferating cell nuclear antigen , cyclins, cyclin-dependent kinases 2 and anti-apoptotic fac-tors were significantly downregulated in MHCC97H cells with FERMT2 knockout (P<0.05).CONCLUSION:FERMT2 may function as a promoter of hepatocarcinogenesis and progression via regulating the cell viability , cell-cycle distribution and cell apoptosis , which is related with the expression of cell cycle regulators and anti-apoptotic factors .