BackgroundThe immune response to hepatitis B vaccine may be influenced by numerous factors, and patients with non/low response re-exposed to hepatitis B virus remain susceptible. Thus, a better understanding of the underlying mechanisms of non/low immune response in infants born to Hepatitis B surface antigen (HBsAg)-positive mothers is essential.Methods100 infants born to HBsAg-positive mothers from 2015 to 2020 were enrolled in the study, further divided into the non/low response group (n=13) and the moderate strong response group (n=87) based on the quantification of hepatitis B surface antibody at 12 months of age. The differential expression of 48 immune-related cytokines in the two groups was compared and analyzed in detail. The key cytokines were further identified and clinically predictive models were developed.ResultsWe found that 13 cytokines were lowly expressed and one cytokine was highly expressed in the non/low response group, compared with the moderate strong response group at birth. In addition, 9 cytokines were lowly expressed and one cytokine was highly expressed in the non/low response group at 12 months of age. Furthermore, we found that IL-5 and HGF were promising predictors for predicting the immunization response to hepatitis B vaccine in infants, and the combination of the two cytokines showed the best predictive efficiency, with an area under the curve (AUC) value of 0.844.ConclusionThe present study provides a theoretical basis on cytokines for developing and implementing effective immunotherapies against non/low immune response in infants born to HBsAg-positive mothers.

The coexistence of congenital diaphragmatic hernia and esophageal atresia with or without tracheoesophageal fistula is extremely rare; only 36 cases have been reported. We report a case of a preterm male newborn infant with left congenital diaphragmatic hernia, esophageal atresia, and tracheoesophageal fistula and review 27 related cases.

Purpose: As there have been few studies on the effects of the receptor for activated C kinase 1 (RACK1) on gastric cancer (GC), we aimed to explore such effects and the mechanism that may be involved. Patients and methods: Normal gastric epithelial cells and six GC cell lines were used to detect the mRNA expression of RACK1. Overexpressing RACK1 was transfected in HGC27 and MGC803 cells. The effects of overexpressing RACK1 on cell viability, migration, and invasion were determined by cell counting kit-8, wound scratch, and Transwell assay, respectively. The expressions of epithelial-mesenchymal transition (EMT) and Wnt/β-catenin signaling related genes were detected using quantitative real-time PCR or Western blot. Wnt pathway agonist LiCl was added into RACK1 overexpressing GC cells, and then cell viability, migration, and invasion were also detected. Results: RACK1 was downregulated in GC cell lines. Under the circumstance that overexpressing RACK1 was successfully transfected in the two lowest RACK1-expressing GC cells, significant inhibition of cell viability, migration, and invasion, promotion to the mRNA and protein expression of E-cadherin, as well as a decrease in the N-cadherin and Snail expressions could be observed. Overexpressing RACK1 also enhanced the protein level of phosphorylation-β-catenin/β-catenin and attenuated c-Jun protein expression. Additionally, LiCl could partially reverse the inhibitory effects of cell viability, migration and invasion by overexpressing RACK. Conclusion: We found RACK1 possibly inhibited epithelial-mesenchymal transition of GC cells through limitation of the Wnt/β-catenin pathway, thereby suppressing cell migration and invasion; RACK1 could also suppress cell growth.

Matrine, also known as oxymatrine, is an important active ingredient of traditional Chinese herb Sophora flavescens. Recent studies have found that matrine may inhibit multiple tumors through inhibiting the tumor cell proliferation, inducing cell apoptosis, blocking cell cycle, suppressing cell invasion and migration and assisting in the synergy, and attenuation of radiotherapy and chemotherapy. This study mainly investigated the role of matrine in gastric cancer and its possible mechanism. The real-time fluorescence quantitative polymerase chain reaction technique showed that matrine inhibited the proliferation and migration of gastric tumor cells and significantly suppressed the expression of miR-93-5p. The dual-luciferase reporter gene assay indicated that AHNAK was a target gene of miR-93-5p and regulated by miR-93-5p and matrine. The torsion test demonstrated that matrine exerted its role via miR-93-5p while miR-93-5p played a role by targeting AHNAK. Thus, this study found that matrine affected the progression of gastric cancer by inhibiting the function of gastric cancer cells through the possible mechanism of inhibiting miR-93-5p expression to increase the expression level of the downstream target gene AHNAK.

The purpose of this study was to investigate the expression profile of circular RNA (circRNA) in gastric cancer cells MGC-803, SGC-7901 and NCI-N87 with different degrees of differentiation and normal gastric epithelial cells GES-1. High throughput circRNA microarray technique was used to detect the differential expression of circRNA between three kinds of differentiated gastric cancer cells and normal gastric epithelial cells. The interaction of microRNAs (miRNAs) with circRNAs was predicted by bioinformatics software, and circRNA, which might have great significance in gastric cancer, was identified by literature search. The results showed that there were 79 up-regulated circRNAs and 229 down-regulated circRNAs in gastric cancer cells with different degrees of differentiation compared with those in normal gastric epithelial cells. Through bioinformatics software analysis and literature retrieval, it was found that hsa_circ_0001897 was related to the staging and metastasis of gastric cancer, while the miR-150-5p, which combined with it, was closely related to many kinds of digestive tract tumors, such as colon cancer, liver cancer, and cholangiocarcinoma. The miR-16-5p, which has binding sites with hsa_circ_0008106 and hsa_circ_0060456, has been confirmed to be involved in the development of gastric cancer. The above results suggest that hsa_circ_0001897, hsa_circ_0008106 and hsa_circ_0060456 may be closely related to the occurrence and development of gastric cancer by interacting with miRNA.

Aims: This study evaluated the mechanism by which salinomycin-induced autophagy blocks apoptosis in PC-3 prostate cancer cells. Main methods: The anti-cancer effects of salinomycin in PC-3 cells were confirmed by flow cytometry, JC-1 staining and western blotting. Then, the autophagic effects were measured by western blotting, GFP-LC3 puncta formation assay, immunofluorescence staining and electron microscopy. Furthermore, we used lentivirus-mediated shRNA to silence ATG3, ATG5 and ATG7 expression in PC-3 cells to investigate the regulatory mechanisms of salinomycin-induced autophagy. Key findings: Salinomycin could induce apoptosis and autophagy in PC-3 cells. Interestingly, autophagy inhibition could enhance salinomycin-induced apoptosis. We further showed that ATG3, a known critical regulator of autophagy, was down regulated and involved in the inhibition of apoptosis by salinomycin-induced autophagy via the AKT/mTOR signaling axis. Significance: Our data indicated that salinomycin-induced autophagy blocks apoptosis via the ATG3/AKT/mTOR signaling axis in PC-3 cells, which provides new clues for the mechanisms of underlying the anti-cancer effects of salinomycin.

MicroRNAs (miRs) serve a role in promoting and suppressing tumors in various types of malignant cancer, such as cervical cancer. However, the regulatory mechanism of miR-124-3p in cervical cancer remains unclear. In the present study, miR-124-3p was significantly downregulated in cervical cancer tissues and cell lines compared with matching adjacent non-tumor tissues and the normal cervical epithelial cell line End1/E6E7, respectively. Decreased expression of miR-124-3p was associated with advanced cervical cancer and the results of an in vitro study demonstrated that the ectopic expression of miR-124-3p significantly decreased the proliferation, migration and invasion of cervical cancer Caski cells. Furthermore, insulin-like growth factor 2 mRNA binding protein 1 (IGF2BP1) was identified as a novel target of miR-124-3p. Overexpression of miR-124-3p decreased the expression of IGF2BP1, whereas miR-124-3p knockdown promoted IGF2BP1 expression at the post-transcriptional level in Caski cells. Additionally, overexpression of IGF2BP1 attenuated the suppressive effects of miR-124-3p on the proliferation, migration and invasion of Caski cells. IGF2BP1 was upregulated in cervical cancer tissues and cell lines compared with matching adjacent non-tumor tissues and the End1/E6E7 cell line, respectively. Therefore, the present study suggests that miR-124-3p suppresses the growth and metastasis of cervical cancer by directly targeting IGF2BP. Thus, miR-124-3p may be developed as a novel method of treating cervical cancer.

The present study profiled differentially expressed microRNAs (miRs) in gastric cancer cell lines and then investigated miR-7 expression in gastric cancer tissue specimens and the effects of miR-7 on the growth, invasion and metastasis of gastric cancer cells and the underlying molecular events. A microRNA microarray was used to profile differentially expressed miRNAs in human gastric cancer cell lines relative to a normal stomach mucosal epithelial cell line. The miRNA miR-7 was selected for further investigation, which included real-time reverse-transcription PCR (qRT-PCR) analysis of miR-7 levels in different gastric cancer cell lines and tissues and distant non-tumor tissues from patient resections. Cell counting kit-8 (CCK-8), Transwell migration and invasion, and western blot assays were performed to assess tumor cell viability, invasion and gene expression, respectively, after miR-7 transfection. The miRNA microarray profiling revealed 14 upregulated miRNAs (including miR-21, miR-26b and miR-30b) and 19 downregulated miRNAs (including let-7i, miR-7 and miR-622) between gastric cancer and normal cell lines. The qRT-PCR analysis confirmed that reduced miR-7 expression occurred more frequently in poorly and moderately differentiated gastric cancer MGC-803, MKN-45 and SGC-7901 cell lines than in the well-differentiated gastric cancer NCI-N87 cell line, which was consistent with the results for gastric cancer tissues. Expression of miR-7 was downregulated in 86.9% (20/23) of the gastric cancer tissues compared with that in the distant non-tumor tissues. Restoration of miR-7 expression significantly inhibited tumor cell viability, invasiveness and migration when compared with the control cells. Luciferase assay confirmed the epidermal growth factor receptor (EGFR) as a target gene of mR-7, and expression of miR-7 significantly suppressed EGFR expression at both the mRNA and protein levels. The data from the present study demonstrated that reduced miR-7 expression contributes to gastric cancer development and progression. Further study will investigate miR-7 in the regulation of EGFR expression in vitro and in vivo.

AIM: To profile expression of microRNAs (miRNAs) in gastric cancer cells and investigate the effect of miR-374b-5p on gastric cancer cell invasion and metastasis.METHODS: An miRNA microarray assay was performed to identify miRNAs differentially expressed in gastric cancer cell lines (MGC-803 and SGC-7901) compared with a normal gastric epithelial cell line. Upregulation of miR-374b-5p was newly identified and confirmed via quantitative real-time reverse transcription-PCR (qRT-PCR). MGC-803 cells were transfected with a synthesized anti-miR-374b-5p sequence or a control vector using Lipofectamine reagent, or treated with transfection reagent alone or phosphate-buffered saline as controls. Rate of transfection was verified after 48 h by qRT-PCR. Cells were then subjected to transwell migration, wound scratch and cell counting kit-8 assays. A bioinformatic analysis to identify miR-374b-5p target genes was performed using miRanda, PicTar and TargetScan software. A dual luciferase reporter assay was performed to evaluate the influence of miR-374b-5p on target gene activation, and qRT-PCR and Western blot were used to evaluate the levels of target mRNA and protein following transfection with miR-374b-5p antisense oligonucleotides.RESULTS: The microarray profiling revealed down-regulation of 14 (fold change < 0.667; P < 0.05) and upregulation of 12 (fold change > 1.50; P < 0.05) miRNAs in MGC-803 and SGC-7901 cells compared with GES-1 controls. The upregulation of miR-374b-5p (fold change = 1.75 and 1.64 in MGC-803 and SGC-7901, respectively; P < 0.05) was confirmed by qRT-PCR. Compared with the control groups, the restoration of miR-374b-5p expression with anti-miR-374b-5p significantly suppressed the metastasis, invasion and proliferation of MGC-803 cells. The bioinformatic analysis predicted that the 3' untranslated region (UTR) of reversion-inducing cysteine-rich protein with Kazal motif (RECK) contains three miR-374b-5p target sequences. RECK was verified as a target gene in a dual luciferase reporter assay showing that activation of RECK 3'UTR-pmirGLO was increased by co-transfection with miR-374b-5p. Finally, transfection of miR-374b-5p antisense oligonucleotides increased mRNA and protein levels of RECK in MGC-803 cells (P < 0.05).CONCLUSION: These findings indicate that upregulation of miR-374b-5p contributes to gastric cancer cell metastasis and invasion through inhibition of RECK expression. (C) 2014 Baishideng Publishing Group Inc. All rights reserved.

Gastric cancer mainly metastasizes via lymphatic vessels. Thus, it is critical to identify efficacious chemopreventive agents for lymphangiogenesis. The present study was undertaken to explore the effects of rosiglitazone (ROSI) on the growth and lymphangiogenesis of human gastric cancer. We established a model of gastric cancer by subcutaneously inoculating the human gastric cancer cell line SGC-7901 into nude mice. Mice were randomly divided into 4 groups and each group received a different agent by oral gavage. The control group received normal saline and treatment groups received different doses of ROSI once every 2 days. The growth of the tumor in vivo was assessed by measuring tumor volume. After 42 days, the mice were sacrificed and the tumors were removed. H&E staining was used to observe the histomorphological features; immunohistochemistry staining for lymphatic vessel density (LVD) was used to evaluate tumor lymphangiogenesis, RT-PCR was performed to determine the mRNA expression of vascular endothelial growth factor C (VEGF-C) and VEGF receptor-3 (VEGFR-3), and western blotting was used to detect the protein expression of VEGF-C and VEGFR-3. Compared with the control group, all treatment groups had smaller tumor volume and higher tumor growth inhibitory rate every day. The number of typical tumor cells in the control group was higher compared to that in the treatment groups, and the highest level of LVD was found in the control group. Furthermore, both the expression of VEGF-C and VEGFR-3 mRNA and proteins in the control group were significantly higher compared to those in the treatment groups. Markedly, these changes were correlated in a dose-dependent manner with ROSI. These results demonstrated that, through simultaneously blocking the expression of VEGF-C and VEGFR-3, ROSI suppresses lymphangiogenesis. This may represent a powerful therapeutic approach for controlling gastric cancer cell growth and metastasis.

Polyglutamine(PolyQ) diseases comprise a group of inherited neurodegenerative disorders with significant clinical and genetic heterogeneity. Although they share a common mechanism involving dynamic expansion of CAG trinucleotide repeats, their clinical features may vary and there has been no specific treatment. Recently, much attention had been attracted to microRNAs which, as a new type of posttranscription regulatory factor, have proven to significantly affect the progress of PolyQ disease. This review will focus on the roles of microRNAs in the pathogenesis of PolyQ diseases and their potential use for therapy.

Introduction: To explore the mechanisms of rosiglitazone (ROS), a selective peroxisome proliferator-activated receptor gamma ligand, in reversing mitomycin C (MMC) resistance in a human drug-resistant gastric cancer cell line. Methods: The vincristine-resistant human gastric cancer cell line SGC7901/VCR and its parental cell line SGC7901 were treated with ROS, MMC (negative control), cyclosporine A+MMC (positive control) or ROS+MMC. A tetrazolium blue (methyl thiazolyl tetrazolium) assay was used to evaluate the sensitivity to these treatments. Flow cytometry analysis and acridine orange-ethidium bromide (AO-EB) fluorescent staining were used to determine the effects of ROS on MMC-induced apoptosis. Reverse transcription polymerase chain reaction and western blotting were used to measure the expression of multidrug resistant 1 (MDR1), Livin and P-glycoprotein (P-gp). Results: ROS administration dose dependently increased the reversal index in MMC-treated SCG7901/VCR cells. ROS increased apoptosis in SGC7901/VCR cells compared with the blank group and MMC group. ROS+MMC also increased apoptosis in SGC7901/VCR cells compared with other groups (P < 0.05 or P < 0.01). The mRNA expression of MDR1 and Livin and the protein expression of P-gp in SGC7901/VCR cells were significantly higher than those in SGC7901 cells (P < 0.01). However, ROS or ROS+MMC treatment markedly upregulated the mRNA expression of MDR1 and Livin and the protein expression of P-gp in SGC7901/VCR cells (P < 0.01). Conclusions: ROS reverses MMC resistance in human gastric cancer SGC7901/VCR cells by reducing expression of MDR1, Livin and P-gp and increasing apoptosis.

Background: Recent studies demonstrated that celecoxib, a selective inhibitor of cyclooxygenase-2 (COX-2), has anticancer effect. Aims: To investigate the effect of celecoxib on expressions of macrophage migration-inhibitory factor (MIF), tumor suppressor gene protein PTEN and apoptosis-associated protein Caspase-3 in human colon cancer cell line HT-29, and its potential molecular mechanism in the prevention and treatment of colon cancer. Methods: Expression of MIF mRNA in HT-29 cells treated with 25, 50 and 100 μmol/L celecoxib, respectively for 24 hours was determined by reverse transcriptase polymerase chain reaction (RT-PCR), and expressions of MIF, PTEN and Caspase-3 proteins were determined by Western blotting. Results: Expressions of MIF mRNA and protein in HT-29 cells treated with 25-100 μmol/L celecoxib were down-regulated in a dose-dependent manner as compared with normal controls (P<0.05), whereas the expressions of PTEN and Caspase-3 proteins were up-regulated in a dose-dependent manner (P<0.05). Conclusions: The anti-colon cancer effect of celecoxib might be related to the down-regulation of MIF and up-regulation of PTEN and Caspase-3.

Background: Macrophage migration inhibitory factor (MIF) is a multifunctional cytokine, its mechanism in the genesis and development of colorectal cancer has not been clarified. Aims: To investigate whether MIF regulates the proliferation of human colon cancer cells and expression of angiogenesis factor via PI3K/Akt signal pathway. Methods: The cultured human colon cancer cell line HT-29 was treated with RPMI-1640 (control group), recombinant human MIF (rhMIF), PI3K inhibitor LY294002 and LY294002 pretreating added with rhMIF, respectively. Methyl thiazolyl tetrazolium (MTT) assay was used to measure the proliferation of HT-29 cells. The mRNA and protein levels of vascular endothelial growth factor (VEGF) and interleukin-8 (IL-8) in HT-29 cells were detected by reverse transcriptase polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA). Western blotting was performed to determine the expressions of Akt and phosphorylated Akt (p-Akt) proteins. Results: rhMIF could promote cell proliferation, increase the mRNA and protein levels of VEGF and IL-8, and induce phosphorylation of Akt protein in HT-29 cells (P<0.05). When pretreated with LY294002, the above-mentioned changes induced by rhMIF were inhibited significantly. The proliferation of HT-29 cells decreased significantly (P<0.05), no statistically significant differences were found in VEGF, IL-8 mRNA and protein levels, as well as in protein level of phosphorylated Akt between LY294002 plus rhMIF group and the controls. Total Akt protein level was similar among all groups. Conclusions: MIF can induce cell proliferation and angiogenesis factor expression in human colon cancer cells via activation of PI3K/Akt signal pathway.

AIM:To investigate the effects of macrophage migration inhibitory factor (MIF) on proliferation of human gastric cancer MGC-803 cells and expression of cyclin D1 and p27(Kip1) in them, and further determine whether the effects are related to the PI3K/Akt signal transduction pathway.METHODS:Gastric cancer MGC-803 cells were cultured and then treated with 50 microg/L recombinant human MIF (rhMIF) with and without a PI3K inhibitor, LY294002 (25 micromol/L). MTT assay was used to detect the proliferation of MGC-803 cells. Cell cycle was detected by flow cytometry. Expression of cyclin D1 and p27(Kip1) mRNA was by reverse transcription-polymerase chain reaction. Protein expression of phosphorylated Akt (p-Akt), Akt, cyclin D1 and p27(Kip1) was examined by immunocytochemistry and Western blotting.RESULTS:rhMIF significantly stimulated the proliferation of MGC-803 cells and cell cycle progression from G1 phase to S phase in a concentration- and time-dependent manner. After the MGC-803 cells were treated with rhMIF for 24 h, the expression of cyclin D1 was significantly up-regulated compared with the cells not treated with rhMIF at both mRNA and protein levels (0.97 +/- 0.02 vs 0.74 +/- 0.01, P = 0.002; 0.98 +/- 0.05 vs 0.69 +/- 0.04, P = 0.003). The p27(Kip1) was down-regulated but only statistically significant at the protein level. rhMIF significantly increased the expression of p-Akt, which reached the peak at 30 min, but did not affect the expression of Akt. However, LY294002 inhibited all the effects of rhMIF.CONCLUSION:Macrophage MIF increases the proliferation of gastric cancer cells, induces the expression of cyclin D1 at the transcriptional level and inhibits the expression of p27(Kip1) at the post-transcriptional level via the PI3K/Akt pathway.

OBJECTIVE To detect the relations of c-erbB-2 oncogene protein, epidermal growth factor receptor (EGFR) and transforming growth factor-beta1 (TGF-beta1) to the progression or metastasis of pancreatic carcinoma. METHODS Using streptavidinbiotin complex (SABC) method, c-erbB-2 oncongene protein, we examined immunohistochemically EGFR and TGF-beta1 expressions in wax-tissue sections from 10 individuals with normal pancreas (NP), 13 patients with chronic pancreatitis (CP) and 36 patients with pancreatic ductal adenocarcinoma (PC). RESULTS The positive expression rates of c-cerbB-2 oncogene protein, EGFR and TGF-beta1 in the NP, CP and PC groups were 0, 0, 10%; 7.7%, 7.7%, 7.7%; and 41.7%, 50.0%, 44.4%, respectively. The positive expression rates of the three specific proteins increased more significantly in the PC group than in the NP and CP groups (P<0.05). The individual expression of c-erbB-2, EGFR and TGF-beta1 was not related to the age and sex of the patients as well as the site, size and histopathological grade of tumors (P>0.05), but to the clinical stage of tumors (P<0.01). The coexpression rate of the three proteins was 27.8% (10/36). This coexpression in the PC group was correlated with the histopathological grades and clinical stages of tumors (P<0.01). CONCLUSION Detection of c-erbB-2 oncogene protein, EGFR, and TGF-beta1 expressions in pancreatic tissue is helpful to judge the malignancy, progression, and metastasis of PC.