Novel MEI1 mutations cause chromosomal and DNA methylation abnormalities leading to embryonic arrest and implantation failure.

This study presents a case of a female infertile patient suffering from embryonic arrest and recurrent implantation failure. The primary objective was to assess the copy number variations (CNVs) and DNA methylation of her embryos. Genetic diagnosis was conducted by whole-exome sequencing and validated through Sanger sequencing. CNV evaluation of two cleavage stage embryos was performed using whole-genome sequencing, while DNA methylation and CNV assessment of two blastocysts were carried out using whole-genome bisulfite sequencing. We identified two novel pathogenic frameshift variants in the MEI1 gene (NM_152513.3, c.3002delC, c.2264_2268 + 11delGTGAGGTATGGACCAC) in the proband. These two variants were inherited from her heterozygous parents, consistent with autosomal recessive genetic transmission. Notably, two Day 3 embryos and two Day 6 blastocysts were all aneuploid, with numerous monosomy and trisomy events. Moreover, global methylation levels greatly deviated from the optimized window of 0.25-0.27, measuring 0.344 and 0.168 for the respective blastocysts. This study expands the mutational spectrum of MEI1 and is the first to document both aneuploidy and abnormal methylation levels in embryos from a MEI1-affected female patient presenting with embryonic arrest. Given that females affected by MEI1 mutations might experience either embryonic arrest or monospermic androgenetic hydatidiform moles due to the extrusion of all maternal chromosomes, the genetic makeup of the arrested embryos of MEI1 patients provides important clues for understanding the different disease mechanisms of the two phenotypes.

Extended-spectrum ß-lactamase-producing Enterobacteriaceae from ready-to-eat foods: Genetic diversity and antibiotic susceptibility.

Ready-to-eat (RTE) foods are widely marketed in China and are important components of everyday diet. In this study, a total of 2000 RTE food samples were analyzed, 252 (12.60%) of which were positive for Enterobacteriaceae, and 48 were identified as containing extended-spectrum ß-lactamase (ESBL)-producing Escherichia coli isolates. Furthermore, the antimicrobial resistance patterns of these isolates to 14 antimicrobial agents revealed that most isolates were resistant to aminoglycosides and ß-lactam antibiotics. The TEM-type gene was prevalent in our isolates (79.17%). The isolates (n = 48) were classified into three clusters based on the ERIC-PCR results. Forty-eight sequence types were found without duplicates, revealing genetic variation and relatedness among isolates. Thus, the results demonstrated the presence of ESBL-producing Enterobacteriaceae in Chinese RTE foods. The results of this study provide insights into the spread of antibiotic-resistant strains and improve understanding of microbial risks.

PBRM1 presents a potential prognostic marker and therapeutic target in duodenal papillary carcinoma.

BACKGROUND: Due to its rarity, duodenal papillary carcinoma (DPC) is seldom studied as a unique disease and no specific molecular features or treatment guidelines are provided. METHODS: Whole-exome sequencing was performed to gain new insights into the DPC mutation landscape and to identify potential signalling pathways and therapeutic targets. Mechanistically, immunohistochemistry (IHC), immunofluorescence, RNA-seq, ATAC-seq and in vitro cell function experiments were performed to confirm the underlying mechanisms. RESULTS: We described the mutational landscape of DPC for the first time as a group of rare tumours with a high frequency of dysregulation in the chromatin remodelling pathway, particularly PBRM1-inactivating mutations that are significantly higher than duodenal adenocarcinomas and ampullary adenocarcinoma (27% vs. 0% vs. 7%, p < .01). In vitro cell experiments showed that downregulation of PBRM1 expression could significantly promote the cancer progression and epithelial-to-mesenchymal transition via the PBRM1-c-JUN-VIM axis. The IHC data indicated that PBRM1 deficiency (p = .047) and c-JUN expression (p < .001) were significantly associated with poor prognosis. Meanwhile, the downregulation of PBRM1 expression in HUTU-80 cells was sensitive to radiation, which may be due to the suppression of c-JUN by irradiation. CONCLUSIONS: Our findings define a novel molecular subgroup of PBRM1-inactivating mutations in DPC. PBRM1 play an important role in DPC progression and may serve as a potential therapeutic target and prognostic indicator.

The new role of riluzole in the treatment of pancreatic cancer through the apoptosis and autophagy pathways.

Pancreatic cancer is always diagnosed at an advanced stage. Hence, chemotherapy becomes the best choice for patients. Therefore, new anticancer drugs for pancreatic cancer are needed. Riluzole (RIL) is mainly used to treat amyotrophic lateral sclerosis clinically, but many previous studies have shown that RIL could inhibit tumors. However, no report has explored the association between RIL and pancreatic cancer. To validate this association, we performed this study. Our data showed that RIL could induce cytotoxicity, block the cell cycle, and inhibit clone formation, apoptosis, and migration in pancreatic cancer cells. Moreover, we demonstrated that RIL could suppress autophagy. However, more experiments will be needed to validate the reliability of our conclusions. In summary, our data suggest that RIL might provide clues for the development of a treatment for human pancreatic cancer in the future.

IL35 predicts prognosis in gastric cancer and is associated with angiogenesis by altering TIMP1, PAI1 and IGFBP1.

Tumor angiogenesis is required for tumor growth and metastasis. Interleukin-35 (IL35), a member of the IL12 family, is a dimer composed of IL12A and EBV-induced gene 3（EBI3）. Elevated plasma IL35 levels have been reported to be associated with the occurrence and development of tumors. However, the role of IL35 in the angiogenesis of gastric cancer (GC) is still unclear. Here, we report that expression of IL35 is correlated with higher microvessel density, distant metastasis and poor prognosis in GC. Moreover, in vitro tube formation assays were performed to show that IL35 may contribute to the tube formation abilities of human umbilical vein endothelial cells. IL12A was observed to be the dominant subunit in promotion of tube formation. IL12A also inhibited expression of tissue inhibitor of metalloproteinase 1 and enhanced expression of plasminogen activator inhibitor 1 and insulin-like growth factor-binding protein 1 in a GC cell line. In conclusion, our data suggest that IL35 is involved in angiogenesis and is associated with poor prognosis for GC.

Integrative Analysis of the IQ Motif-Containing GTPase-Activating Protein Family Indicates That the IQGAP3-PIK3C2B Axis Promotes Invasion in Colon Cancer.

BACKGROUND: Colon cancer (CRC) is a common type of tumour, and IQGAP family proteins play an important role in many tumours. However, their roles in CRC remain unclear. METHODS: First, we searched many public databases to comprehensively analyze expression of IQGAPs in CRC. Next, real-time PCR, immunohistochemical (IHC), transwell, siRNA transfection and Western blot assays were used to evaluate relationships among IQGAP3 expression, clinical pathological parameters and CRC prognosis, and the underlying molecular mechanism was investigated. RESULTS: IQGAP3 was elevated in CRC tissues, whereas there was no significant change in expression of IQGAP1 or IQGAP2. Additionally, IQGAP3 expression in CRC tissues was associated with tumour progression, invasion and poor prognosis. In mechanistic studies, we found that IQGAP3 was positively coexpressed with PIK3C2B. In an in vitro assay, the PIK3C2B expression level was increased after exogenous overexpression of IQGAP3, resulting in the promotion of cell invasion, which was blocked by pretransfecting cells with PIK3C2B siRNA. Furthermore, we found that high expression of IQGAP3 and PIK3C2B correlated with tumour stage and vessel invasion in human CRC, whereby patients with high expression of both in tumours had a worse prognosis compared with patients with single-positive or double-negative tumours. CONCLUSION: The results of our current study and corresponding previous studies provide evidence that IQGAP3 is elevated in CRC and promotes colon cancer growth and metastasis by regulating PIK3C2B activation.

Correction: Cu2O template synthesis of high-performance PtCu alloy yolk-shell cube catalysts for direct methanol fuel cells.

Correction for 'Cu2O template synthesis of high-performance PtCu alloy yolk-shell cube catalysts for direct methanol fuel cells' by Sheng-Hua Ye et al., Chem. Commun., 2014, 50, 12337-12340, DOI: 10.1039/C4CC04108A.

Prevalence, antibiotic susceptibility and characterization of Vibrio parahaemolyticus isolates in China.

Vibrio parahaemolyticus is a marine and estuarine bacterium that poses a major threat to human health worldwide. In this study, from 2017 to 2019, we evaluated 900 food samples collected from China in 2017, with the aim of determining the incidence and features of V. parahaemolyticus in ready-to-eat (RTE) foods, shrimp and fish in China. The contamination rates in these were 3.67, 19.33 and 10.67%, respectively, and the prevalence of V. parahaemolyticus was higher in summer than in winter. In addition, 101 V. parahaemolyticus strains were isolated. Our results suggested that most of the isolates were resistant to aminoglycosides based on the antimicrobial resistance patterns of these aquatic product isolates against 14 antimicrobial agents. Furthermore, most of the isolates were multidrug-resistant. Serotyping showed that the isolates of the O2 serotype comprised the maximum proportion. Enterobacterial repetitive intergenic consensus sequence (ERIC)-PCR results indicated that the isolates (n = 101) could be classified into 12 clusters. There were 82 STs suggesting genetic variation and relatedness among these isolates. Our findings demonstrated the presence of V. parahaemolyticus in foods from Chinese retail markets and show that this methodology can be used for microbiological risk assessment in China.

Advantages of Restoring miR-205-3p Expression for Better Prognosis of Gastric Cancer via Prevention of Epithelial-mesenchymal Transition.

PURPOSE: miR-205 is a tumor suppressor and plays an important role in tumor invasiveness. However, the role of miR-205 in human gastric cancer (GC) epithelial-mesenchymal transition (EMT) remains unclear. The aim of this study was to investigate the molecular mechanism of miR-205 in the regulation of EMT in GC invasion. MATERIALS AND METHODS: Quantitative polymerase chain reaction (qPCR) was used to detect the expression of miR-205 in GC. Further, the correlation between the pathological parameters and prognosis of GC was statistically analyzed. A transwell model was used to evaluate the effect of miR-205-3p on the invasion and migration of GC cells. qPCR, western blotting, and luciferase assay were performed to analyze the relationship and target effects between miR-205-3p and the expression of zinc finger electron box binding homologous box 1 (ZEB1) and 2 (ZEB2). RESULTS: We found that the levels of miR-205-3p were significantly lower (P<0.05) in GC tissues than in matched normal tissues. Additionally, the expression of miR-205-3p was related to the tumor invasion depth, lymph node metastasis, lymph node invasion, and tumor, node, metastasis stage. Patients with lower miR-205-3p expression levels in the tumors had a poorer prognosis. The in vitro assays indicated that miR-205-3p could affect the invasion ability and EMT of GC cells by targeting the expression of both ZEB1 and ZEB2. CONCLUSIONS: miR-205-3p promotes GC progression and affects the prognosis of patients by targeting both ZEB1 and ZEB2 to directly influence EMT.

Upregulation of miRNA­301a­3p promotes tumor progression in gastric cancer by suppressing NKRF and activating NF­κB signaling.

MicroRNA­301a (miRNA/miR­301a) and nuclear factor (NF)­κB signaling play important roles in tumor invasion, migration and progression. However, the role of miRNA­301a­3p in human gastric cancer (GC), and specifically in the activation of NF­κB signaling, remains unclear. The aim of the present study was to investigate miRNA­301a­3p expression in GC progression and the molecular mechanisms as regards the regulation of NF­κB signaling. Reverse transcription­quantitative polymerase chain reaction (RT­qPCR) was used to detect miRNA­301a­3p expression in GC and paired normal tissues. The association between the expression of miRNA­301a­3p and patient pathological parameters and the prognosis of GC was statistically analyzed using an in situ hybridization (ISH) assay. An MTS assay and a Transwell assay were performed to evaluate the effects of miRNA­301a­3p on the proliferation, invasion and migration of GC cells. RT­qPCR and western blot analysis were used to analyze the association between miRNA­301a­3p and nuclear factor­κB repressing factor (NKRF) expression and the corresponding downstream NF­κB signaling molecules. A luciferase assay was used to verify the target effect of miRNA­301a­3p and NKRF. It was found that miRNA­301a­3p expression was significantly higher in 30 cases of primary GC compared with matched normal tissues. Additionally, the ISH assay indicated that the high expression of miRNA­301a­3p in GC was associated with tumor invasion depth, lymph node metastasis, lymph node invasion and tumor metastasis stage. Patients whose tumors had a higher miRNA­301a­3p expression level exhibited a poorer prognosis. The in vitro assay indicated that miRNA­301a­3p affected the proliferative and invasive ability of GC cells by targeting the expression of NKRF, which then affected NF­κB signaling. Therefore, it was hypothesize that miRNA­301a­3p promotes GC progression and affects the prognosis of patients with GC by targeting NKRF, which in turn, directly influences NF­κB activation.

COX6B2 drives metabolic reprogramming toward oxidative phosphorylation to promote metastasis in pancreatic ductal cancer cells.

Pancreatic ductal adenocarcinoma (PDAC) is an aggressive cancer type with poor prognosis due to its high metastatic potential, however, the role of metabolic reprogramming in the metastasis of PDAC cell is not known. Here, we report that COX6B2 drive metastasis but not cancer cell proliferation in PDAC by enhancing oxidative phosphorylation function (OXPHOS). Transcriptome and clinical analyses revealed that cytochrome c oxidase subunit 6B2 (COX6B2) positively associated with metastasis of PDAC cells. Knockdown of COX6B2 in PDAC cells tuned down the assembly of complex IV and downregulated the function of OXPHOS, whereas re-expression of COX6B2 restored the function of OXPHOS and metastatic potential. Mechanistically, COX6B2 upregulated OXPHOS function to active purinergic receptor pathway for the metastasis of PDAC cells. Notably, the metastatic potential in PDAC could be reversely regulated by metformin, a drug was found accelerating the degradation of COX6B2 mRNA in this study. Collectively, our findings indicated that a complex metabolic control mechanism might be involved in achieving the balance of metabolic requirements for both growth and metastasis in PDAC, and regulation of the expression of COX6B2 could potentially encompass one of the targets.

Bifidobacterium infantis Relieves Allergic Asthma in Mice by Regulating Th1/Th2.

BACKGROUND Bifidobacteria are among the probiotics used in treating intestinal diseases and are rarely used for allergic asthma treatment. The present study investigated the mechanism of B. infantis in treating allergic asthma in mice. MATERIAL AND METHODS A total of 40 male Balb/c mice were randomized into control, ovalbumin (OVA), montelukast (Mon), and B. infantis (B10) groups, and allergic asthma was induced in the OVA, Mon, and B10 groups. Airway reactivity was measured on day 29 by methacholine at various doses. The numbers of total cells and inflammatory cells in bronchoalveolar lavage fluid (BALF) were counted by blood cell counter and Diff-Quik staining. Hematoxylin-eosin (HE) staining was performed to observe inflammatory cell infiltration in lung tissues. Total IgE and OVA-specific IgE in serum were measured by ELISA. Mucin 5AC expression was detected by Western blot to evaluate airway obstruction. The levels of Th1 (IFN-γ, IL-2) and Th2 (IL-4, IL-5, IL-13) cytokines in BALF and tissues were detected by ELISA and qRT-PCR, respectively. RESULTS The mice in the OVA group had airway hyperreactivity, while the symptoms in the B10 group and Mon group were effectively relieved. B10 reduced the number of inflammatory cells in BALF as well as inflammatory cell infiltration in tissues. Moreover, the levels of total serum IgE, OVA-specific IgE, and Mucin 5AC were increased in the OVA group, but were reduced in the Mon group and B10 group. B. infantis increased the levels of Th1 cytokines and decreased those of Th2 cytokines. CONCLUSIONS B. infantis can reduce the infiltration of inflammatory cells induced by OVA-specific antibodies in mice. B. infantis has therapeutic effects on allergic asthma by promoting Th1 and inhibiting Th2 immune responses.

4-Hydroxyphenylpyruvate Dioxygenase-Like Protein Promotes Pancreatic Cancer Cell Progression and Is Associated With Glutamine-Mediated Redox Balance.

Tumor cells develop a series of metabolic reprogramming mechanisms to meet the metabolic needs for tumor progression. As metabolic hubs in cells, mitochondria play a significant role in this process, including energy production, biosynthesis, and redox hemostasis. In this study, we show that 4-hydroxyphenylpyruvate dioxygenase-like protein (HPDL), a previously uncharacterized protein, is positively associated with the development of pancreatic ductal adenocarcinoma (PDAC) and disease prognosis. We found that overexpression of HPDL in PDAC cells promotes tumorigenesis in vitro, whereas knockdown of HPDL inhibits cell proliferation and colony formation. Mechanistically, we found that HPDL is a mitochondrial intermembrane space localized protein that positively regulates mitochondrial bioenergetic processes and adenosine triphosphate (ATP) generation in a glutamine dependent manner. Our results further reveal that HPDL protects cells from oxidative stress by reprogramming the metabolic profile of PDAC cells toward glutamine metabolism. In short, we conclude that HPDL promotes PDAC likely through its effects on glutamine metabolism and redox balance.

[Expression of tumor-associated vascular insulin receptor in colorectal cancer and its relationship with tumor pathological features].

OBJECTIVE: To study the expression of tumor associated vascular insulin receptor (TVIR) in colorectal cancer with or without metabolic syndrome (MS) and its relationship with the pathological features of colorectal cancer. METHODS: The expression of TVIR in 220 colorectal cancer specimens was detected by tissue microarray and immunohistochemistry. The relationships between the expression of TVIR and the pathological features (pathological subtypes, histological grade, invasion depth, lymph node metastasis and TNM stage) of colorectal cancer with/without MS were analyzed. RESULTS: The insulin receptor expression was observed in colorectal cancer tissue or border area between cancer and normal tissue, but not in normal intestinal tissue. The high-expression rates of TVIR in MS group was remarkably lower than that of non-MS group (21.6%vs. 41.0%, P< 0.05). TVIR high expression was significantly associated with tumor deep invasion, lymph node metastasis and high TNM stage (P<0.05 or P<0.01). Univariate logistic regression analysis showed high-expression of TVIR to be significantly associated with risk of deep invasion (OR=2.21, 95% CI: 1.10-4.44, P<0.05), lymph node metastasis (OR=2.21, 95% CI: 1.26-3.86, P<0.01) and high TNM stage (OR=2.08, 95% CI: 1.19-3.63, P<0.05). Such associations can be observed in patients without MS, but not in patients with MS. CONCLUSIONS: s: High-expression of TVIR is associated with aggressive pathological features such as invasion, lymph node metastasis and high TNM stage of colorectal cancer, especially for those patients without MS. TVIR could be a useful biological marker for prognosis of colorectal cancer.

Antimicrobial resistance and genetic diversity of Salmonella enterica from eggs.

Salmonella enterica is a common foodborne pathogen responsible for major global health problems such as paratyphoid fever and gastroenteritis. Here, we report the prevalence, antibiotic resistance phenotypes, serotypes, and molecular subtyping of Salmonella isolated from eggs in Guangdong, China. Out of 1,000 egg samples, 54 (5.40%) were positive. S. Enteritidis made up the largest proportion of samples with 11 serotypes. Antimicrobial susceptibility test indicated that most strains were resistant to ß-lactam, aminoglycoside, and tetracycline antibiotics (27.00%-40.00%). There were 37 STs based on MLST typing. MLST and ERIC-PCR classified 54 isolates into three and five clusters, respectively, which revealed the genetic relatedness and diversity. In conclusion, frequent monitoring of eggs for Salmonella, antibiotic resistance profiles and genetic diversity is essential for improving food safety.

Tumor-associated macrophages induce invasion and poor prognosis in human gastric cancer in a cyclooxygenase-2/MMP9-dependent manner.

Cyclooxygenase-2 (COX2) and tumor-associated macrophages (TAMs) are associated with invasion, angiogenesis, and poor prognosis in many human cancers. However, the role of TAMs in human gastric cancer (GC) remains elusive. In the present study, we first measured COX2 expression and TAM infiltration in human GC tissues using double immunohistochemical staining. Then, we indirectly cocultured M2-polarized macrophages derived from human THP-1 cells with GC cells as an in vitro model. Transwell assays, siRNA transfection, treatment with a COX2 inhibitor and Western blotting were used to investigate the relationship among TAMs, invasion and COX2 expression as well as the underlying molecular mechanism. Double IHC staining showed that TAMs were aggregated near GC tumor nests and had high COX2 expression; moreover, the number of TAMs that infiltrated the tumor nest was correlated with the depth of invasion, COX2 expression and poor prognosis in human GC. In an in vitro assay, after treatment with phorbol myristate acetate (PMA), the THP-1 cells differentiated into M2 macrophages and induced COX2/MMP9-dependent invasiveness in GC cells. Pretreatment of GC cells with COX2 siRNA or a COX2 inhibitor (Celecoxib) can negate these promoting effects. The results of this study and those of our previous studies indicate that coculture with M2-polarized macrophages can induce the COX2-dependent release of matrix metalloproteinase-9 (MMP9), which subsequently increases the invasiveness of GC cells. Our data may provide a basis for targeting TAMs or for polarizing TAMs through immune regulation to halt GC progression, which could soon become a nonsurgical treatment for human gastric cancer.

Breast cancer prognosis is better in patients who develop subsequent metachronous thyroid cancer.

Breast cancer (BC) and thyroid cancer (TC) are common malignancies among females. However, the connection between TC and BC is not well understood. To explore the relationship between these two cancers and to determine the effect of second metachronous TC on BC survival, we compared BC patients with or without second primary TC using data from the Surveillance, Epidemiology, and End Results (SEER) database. We extracted data from patients with only BC or TC and from BC patients with a second metachronous cancer from 2000-2014. Differences in the clinicopathological and treatment characteristics between BC patients with or without second metachronous TC were analyzed by chi-square tests. Multivariate analyses of BC survival were performed by using Cox regression models. Comparison of disease-specific survival (DSS) curves between these cohorts was performed with the log-rank (Mantel-Cox) test. Survival analyses were also performed using data from 1980-1994. Within this dataset, we found 1,262 BC cases in which a second metachronous TC (BC2TC) developed, accounting for 3.1% of all metachronous cancers following BC from 2000-2014. No significant differences were found in molecular markers. In addition, the mean age at BC diagnosis was younger in the BC2TC group than in the BC group (55.418 y vs 60.273 y). Half of the BC2TC patients developed TC in the first three years following BC diagnosis. Patients with BC2TC showed better DSS than those with BC alone from 2000-2014 (P<0.001). However, this superiority was not significant from 1980-1994 (P = 0.579) or for TNM stage I BC (P = 0.927) and grade I BC (P = 0.431) from 2000-2014. In conclusion, the incidence of BC2TC has increased dramatically during the past 15 years. In addition, patients with BC2TC showed better DSS than patients with BC alone, especially in cases from 2000-2014.

Prevalence and genetic diversity of Vibrio parahaemolyticus strains from salmon in Chinese markets.

Consumption of contaminated salmon in China has led to pathogenic Vibrio parahaemolyticus infections in humans. In this study, 420 salmon samples were collected from supermarkets and restaurants in China that showed a contamination rate of 9.05 and 15.24%, respectively. Eighteen antibiotics were used to test the antibiotic susceptibility of all 51 isolates. The most common resistance was observed to ß-lactam antibiotics and aminoglycosides, including ampicillin (92.16%), streptomycin (88.24%), kanamycin (45.10%) and cephazolin (45.10%). Meanwhile, many strains were resistant to more than two antibiotics (48/51, 94.12%). Only two and five isolates were positive for tdh and trh, respectively. Serotyping results demonstrated that O2 was most prevalence serotype (15/51, 29.41%). Molecular typing (enterobacterial repetitive intergenic consensus sequence polymerase chain reaction and multilocus sequence typing) allowed classification of all the isolates into 5 clusters and 44 sequence types, highlighting genetic variation and relatedness. In general, the high antibiotic resistance is alarming and raises public health concerns. Frequent monitoring of salmon for V. parahaemolyticus contamination, genetic diversity and antibiotic susceptibility is essential to improve seafood safety.