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Supplementary Figures S1-S6 and Figure Legends. Figure S1. Western blotting and RT-PCR results depicting CDK5 expression levels from 171 patients. Figure S2. Immunofluorescence staining of CDK1, CDK2, CDK3, and CDK4 in gastric cells. Figure S3. Nuclear/cytoplasmic distribution of CDK5 during cell cycle activity in gastric cancer cells. Figure S4. Flow Cytometry assay for identification of synchronized gastric cancer cells. Figure S5. Gastric cancer cells overexpressing nuclear CDK5 or CDK5-KD cells exhibited a much retarded in vitro growth rate than control cells. Figure S6. NS-0011 induced the nuclear translocation of CDK5 in MGC-803 cells by dose dependent.

Supplementary Tables S1-S3 and Table Legends. Table S1. Clinicopathological parameters of gastric cancer patients with Low or High CDK5 Expression. Table S2. Univariate analysis of the correlation between clinicopathological parameters and survival of patients with gastric cancer. Table S3. Multivariate Analysis of the Correlation between Clinicopathological Parameters and Survival Time of Patients with Gastric Cancer.

Purpose: As a cyclin-independent atypical CDK, the role of CDK5 in regulating cell proliferation in gastric cancer remains unknown.Experimental Design: Expression of CDK5 in gastric tumor and paired adjacent noncancerous tissues from 437 patients was measured by Western blotting, immunohistochemistry, and real-time PCR. The subcellular translocation of CDK5 was monitored during gastric cancer cell proliferation. The role of nuclear CDK5 in gastric cancer tumorigenic proliferation and ex vivo xenografts was explored. Furthermore, by screening for compounds in the PubChem database that disrupt CDK5 association with its nuclear export facilitator, we identified a small molecular (NS-0011) that inhibits gastric cancer cell growth.Results: CDK5 level was significantly decreased in the majority of gastric tumor tissues, and the reduction of CDK5 correlated with the severity of gastric cancer based on tumor and lymph node metastasis and patient 5-year fatality rate. Nuclear localization of CDK5 was found to be significantly decreased in tumor tissues and gastric cancer cell lines, whereas exogenously expression of nucleus-targeted CDK5 inhibited the proliferation and xenograft implantation of gastric cancer cells. Treatment with the small molecule NS-0011, which increases CDK5 accumulation in the nucleus, suppressed both cancer cell proliferation and xenograft tumorigenesis.Conclusions: Our results suggest that low CDK5 expression is associated with poor overall survival in patients with gastric cancer, and nuclear accumulation of CDK5 inhibits the proliferation and tumorigenicity of human gastric cancer cells. Clin Cancer Res; 21(6); 1419–28. ©2015 AACR.

Background: Gastric cancer (GC) ranks fifth in incidence and fourth for mortality worldwide. The response to immune checkpoint blockade (ICB) therapy in GC is heterogeneous due to tumor-intrinsic and acquired immunotherapy resistance. Methods: A novel immunophenotype-based subtyping algorithm was established to analyze tumor patient-derived bulk transcriptomic data. Gene set enrichment assays (GSEA), in vitro cell models, multiple syngeneic murine tumor models, and immune-checkpoint blockade (ICB) therapy were utilized. Results: Our algorithm stratified human GC and showed that immune desert- and excluded-type tumors are ICB-resistant compared with immune-inflamed GC. Moreover, epithelial-mesenchymal transition (EMT) signaling is highly enriched in immune desert-type GC, and syngeneic murine tumors exhibiting mesenchymal-like, compared with epithelial-like, properties are T cell-excluded and resistant to CTLA4 blockade. Our analysis further identifies a panel of receptor tyrosine kinases (RTKs) as potential druggable targets in the immune desert-type GC. Dovitinib, an inhibitor of multiple RTKs, strikingly repressed EMT programming in mesenchymal-like immune desert syngeneic GC models. Dovitinib activates the tumor-intrinsic SNAI1/2-IFN-γ signaling axis and impedes the EMT program converting immune desert-type tumors to immune inflamed-type tumors, sensitizing these mesenchymal-like ‘cold’ tumors to CTLA4 blockade. Conclusion: We develop a novel immunophenotype-based algorithm to reclassify GC into immune inflamed, excluded, and desert subtypes. Our analysis identified major signaling pathways and potential druggable targets relevant to patient groups, especially for refractory immune desert-type/‘cold’ tumors. Dovitinib, an RTK inhibitor, sensitizes desert-type immune-cold GCs to CTLA4 blockade and possibly in a broad spectrum of other solid tumors. Citation Format: Heng Lu, Longlong Cao, Mohammed Soutto, Nadeem Bhat, Zheng Chen, Dunfa Peng, Ahmed Gomaa, Sachiyo Nomura, Jashodeep Datta, Alexander Zaika, Wael El-Rifai. Dovitinib restricts EMT and promotes T cell recruitment to immune-desert gastric cancers via tumor-intrinsic IFN-γ signaling [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3237.

The incidence of esophageal adenocarcinoma (EAC) has increased rapidly over the past four decades. Chronic gastroesophageal reflux disease (GERD), where acidic bile salts (ABS) abnormally refluxate into the esophagus, is the leading risk factor for the development of a metaplastic condition known as Barrett’s esophagus (BE) and its progression to EAC. We used 3D organotypic culture, mouse and human tissue samples to assess the role of ABS in Epithelial-to-Mesenchymal Transition (EMT) in EAC. Analysis of public databases revealed significant enrichment of EMT signaling in EAC progression. RNA-seq analysis of EAC cells showed activation of EMT pathway in response to ABS exposure. ABS induced multiple characteristics of the EMT process, such as downregulation of E-cadherin, upregulation of Vimentin, activation of ß-catenin and EMT-transcription factors (EMT-TFs), cell morphological changes, and enhancement of cell migration and invasion capabilities. Mechanistically, we discovered that ABS induced E-cadherin cleavage via MMP14 proteolytic cascade. APE1 silencing, or APE1-redox-specific inhibitor (E3330), abrogated the ABS-induced EMT process and signaling by downregulating MMP14. Furthermore, APE1 and MMP14 co-expression levels were inversely correlated with E-cadherin expression in gastroesophageal junctions of human EAC tissues, and the L2-IL1ß transgenic mouse model of BE/EAC. EAC patients with high APE1 and EMT signatures had worse relapse free survival than those with low signature. In a summary, this study demonstrates the role of ABS in promoting EMT via the redox-sensitive signaling axis of APE1/MMP14/E-cadherin/ß-catenin. Pharmacological inhibition of APE1-redox function could be a potential therapeutic approach to effectively reduce the risk of Barrett’s carcinogenesis. Citation Format: Heng Lu, Longlong Cao, Farah Ballout, Abbes Belkhiri, Dunfa Peng, Lei Chen, Mohammed Soutto, Oliver McDonald, Alexander Zaika, Jianwen Que, Wael El-Rifai. Gastroesophageal reflux disease promotes E-cadherin cleavage and activates EMT via APE1-redox function in esophageal adenocarcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 1232.

Background: Cell cycle dysregulation is a hallmark of cancer, resulting in unregulated cell proliferation and, eventually, tumor development. Cyclin-dependent kinase 1 (CDK1) is a cell cycle regulatory protein that is involved in cell cycle maintenance. CDK1 has been discovered to be substantially elevated in a several tumors and is linked to poor overall and relapse-free survival. The aim of this study is to understand the regulation role of Helicobacter Pylori infection and inflammation on CDK1 expression in gastric cancer. Methods: Using TCGA data and our integrated comprehensive gene expression analysis, we found a significant overexpression of CDK1 in gastric cancer human and mouse tissues. We detected overexpression of CDK1 in human and mouse gastric glands in response to H. pylori infection. Our data demonstrated that H. pylori infection induced phosphorylation (S536) and activation of NF-kB in vitro and in vivo. Furthermore, H. pylori infection and TNF-α treatment increased the CDK1 mRNA and protein levels in gastric cancer cell lines. Using the ChIP assay, we detected direct biding of NF-κB on the CDK1 promoter regulating its transcription. CDK1 promoted activation of the β-catenin signaling pathway. Using the pTOP/pFOP luciferase reporter assays, as a measure of β-catenin/TCF transcription activity, we confirmed CDK1-dependent activation of β-catenin in response to H. pylori infection. Pharmacologic and genetic inhibition of CDK1 reversed these effects and decreased number and size of gastric tumors organoid from mouse and human. Conclusion: Our findings demonstrate NF-kB-mediated induction of CDK1 expression in response to H. pylori infection with subsequent activation of tumorigenic β-catenin signaling. This novel regulatory link between infection, CDK1, and β-catenin suggests the importance of considering CDK1 inhibitors in gastric cancer. Citation Format: Marwah M. Al-Mathkour, Shoumin Zhu, Longlong Cao, Shayan Khalafi, Zheng Chen, Julio Poveda, Dunfa Peng, Heng Lu, Mohammed Soutto, Tianling Hu, Oliver McDonalnd, Alexander Zaika, Wael El-Rifai. CDK1 bridges NF-kB and b-catenin signaling in response to H. pylori infection in gastric tumorigenesis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 5972.

Background Porcine circovirus type 2 (PCV2) is one of the crucial swine viral pathogens, caused porcine circovirus associated diseases (PCVAD). Shandong province is one of the most important pork producing areas and bears a considerable economic loss due to PCVAD. However, there is limited information on epidemiology and coinfection rate of PCV2 with other critical swine diseases in this area, such as porcine reproductive and respiratory syndrome virus (PRRSV), classical swine fever virus (CSFV), Pseudorabies virus (PRV), and porcine epidemic diarrhea virus (PEDV). Results Overall, 89.59% serum samples and 36.98% tissue samples were positive for PCV2 specified ELISA and PCR positive for PCV2, respectively. The coinfection rates of PCV2 with PRRSV, PRV, CSFV, and PEDV were 26.73%, 18.37%, 13.06%, and 3.47%, respectively. Moreover, genetic characteristic of PCV2 were analyzed based on the cap genes showing that PCV2d is the dominant sub-genotype circulating in the province. Conclusions Our findings reveal that PCV2d, as the dominant strain, is prevailing in pig farms in Shandong province at high levels. There was a high frequency of coinfection of PCV2 and PRRSV.

Inhibition of the immune microenvironment is the main cause of tumor recurrence after surgery in patients with gastric cancer (GC). In this study, immunohistochemistry and multiple immunofluorescence staining were used to evaluate immunosuppressive indicators and immune biomarkers in 825 patients with gastric cancer from three centers. We constructed an immunosuppressive recurrence score (IRS) using LASSO Cox regression based on the expression of six immunosuppressive indicators and found that the IRS and IRS-based nomogram were significantly accurate and reliable in predicting recurrence. Moreover, an elevated IRS was associated with locoregional recurrence and postoperative adjuvant chemotherapy failure. Furthermore, an increase in IRS indicated inhibition of the antitumor effect of CD8(+) tumor-infiltrating lymphocytes in the invasive margin. Thus, we propose that the IRS can predict the recurrence outcome of patients with GC by distinguishing the immunosuppressive status, which is helpful in the selection of individualized adjuvant treatment plans.

Background Currently, pathological examination of gastroscopy biopsy specimens is the gold standard for gastric cancer (GC) diagnosis. However, it has a false-negative rate of 10–20% due to inaccurate sampling locations and/or insufficient sampling amount. A signature should be developed to aid the early diagnosis of GC using biopsy specimens even when they are sampled from inaccurate locations. Methods We extracted a robust qualitative transcriptional signature, based on the within-sample relative expression orderings (REOs) of gene pairs, to discriminate both GC tissues and adjacent-normal tissues from non-GC gastritis, intestinal metaplasia and normal gastric tissues. Results A signature consisting of two gene pairs for GC diagnosis was identified and validated in data of both biopsy specimens and surgical resection specimens pooled from publicly available datasets measured by different laboratories with different platforms. For gastroscopy biopsy specimens, 96.20% of 79 non-GC tissues were correctly identified as non-GC, and 96.84% of 158 GC tissues and six of seven adjacent-normal tissues were correctly identified as GC. For surgical resection specimens, 98.37% of 2560 GC tissues and 97.28% of 221 adjacent-normal tissues were correctly identified as GC. Especially, 97.67% of the 257 GC patients at stage I were exactly diagnosed as GC. We additionally measured 21 GC tissues from seven different GC patients, each with three specimens sampled from three tumor locations with different proportions of the tumor epithelial cell. All these GC tissues were correctly identified as GC, even when the proportion of the tumor epithelial cell was as low as 14%. Conclusions The qualitative transcriptional signature can distinguish both GC and adjacent-normal tissues from normal, gastritis and intestinal metaplasia tissues of non-GC patients even using inaccurately sampled biopsy specimens, which can be applied robustly at the individual level to aid the early GC diagnosis. Electronic supplementary material The online version of this article (10.1186/s12967-019-1816-4) contains supplementary material, which is available to authorized users.

Esophageal adenocarcinoma (EAC) is one of the most lethal of all human malignancies. EAC arises in the setting of Barrett’s esophagus, which is an intestinal metaplastic precursor lesion that develops from chronic reflux of gastrointestinal contents (especially acidic bile salts). Here we report that NOTCH signaling is activated in the esophagus through a unique pathway during exposure to acidic bile salts and progression to EAC. Using a combination of public databases, EAC cell line models, transgenic mice, and patient tissue samples, we detected significant upregulation of several NOTCH signaling components in EAC. Activated NOTCH signaling was confirmed by nuclear accumulation of NOTCH1 cleaved fragment (NICD) with corresponding up-regulation of NOTCH targets in EAC cells in response to acidic bile salts. Moreover, we identified DLL1 as the predominant ligand contributing NOTCH1 activation. Remarkably, DLL1 was regulated by direct cross talk between redox and inflammatory pathways that are activated during both reflux and malignant transformation. Mechanistically, the APE1 redox function transcriptionally up-regulated NF-κB in response to bile salts. This licensed NF-κB to transcriptionally up-regulate DLL1 to activate and stimulate downstream NOTCH1 signaling, thereby defining a novel APE1-NF-kB-NOTCH pro-tumorigenic pathway. This pathway was important for maintaining tumor initiating (cancer “stem cell-like”) properties in vitro, recurrently detected in genetically engineered mouse models of EAC and in EAC patient samples in vivo, and portended an overall poor prognosis. Collectively, these findings indicate that progression from chronic injury to malignancy in the esophagus is driven by a unique mechanism that links redox balance, inflammation, embryonic development (NOTCH) together into a common pro-tumorigenic pathway that is intrinsic to EAC cells. Citation Format: Lei Chen, Heng Lu, Dunfa Peng, Longlong Cao, Zheng Chen, Ajaz Bhat, Alexander Zaika, Shutian Zhang, Wael El-Rifai. Activation of NOTCH signaling via DLL1 is mediated by APE1-redox-dependent NF-κB activation in esophageal adenocarcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 6071.

Helicobacter pylori infection is the predominant risk factor for gastric cancer. RAS protein activator like 2 (RASAL2) is considered a double-edged sword in carcinogenesis. Herein, we investigated the role of RASAL2 in response to H pylori infection and gastric tumorigenesis.Bioinformatics analyses of local and public databases were applied to analyze RASAL2 expression, signaling pathways, and clinical significance. In vitro cell culture, spheroids, patient-derived organoids, and in vivo mouse models were used. Molecular assays included chromatin immunoprecipitation, co-immunoprecipitation, Western blotting, quantitative polymerase chain reaction, and immunocyto/histochemistry.H pylori infection induced RASAL2 expression via a nuclear factor-κB (NF-κB)-dependent mechanism whereby NF-κB was directly bound to the RASAL2 promoter activating its transcription. By gene silencing and ectopic overexpression, we found that RASAL2 upregulated β-catenin transcriptional activity. RASAL2 inhibited protein phosphatase 2A activity through direct binding with subsequent activation of the AKT/β-catenin signaling axis. Functionally, RASAL2 silencing decreased nuclear β-catenin levels and impaired tumor spheroids and organoids formation. Furthermore, the depletion of RASAL2 impaired tumor growth in gastric tumor xenograft mouse models. Clinicopathological analysis indicated that abnormal overexpression of RASAL2 correlated with poor prognosis and chemoresistance in human gastric tumors.These studies uncovered a novel signaling axis of NF-κB/RASAL2/β-catenin, providing a novel link between infection, inflammation and gastric tumorigenesis.

Simple Summary Esophageal adenocarcinoma has become a major clinical challenge in the western world due to its rapid increasing incidence and poor overall prognosis. Understanding the molecular events of its tumorigenesis is the key to better diagnosis and development of better therapeutic strategies. In the current study we aimed to identify epigenetic alteration targets in esophageal adenocarcinoma. We focused on a candidate gene, NDRG4 (N-myc downregulated gene 4). We found that NDRG4 was frequent downregulated in esophageal adenocarcinoma through DNA hypermethylation of its promoter region. Re-expression of NRDG4 in cancer cells significantly suppressed tumor growth via inhibition of cell proliferation. These results will improve our understanding on how dysfunction of NDRG4 contributes to esophageal adenocarcinoma. DNA hypermethylation of NDRG4 may be a useful biomarker in clinical monitoring of esophageal adenocarcinoma patients. Abstract The incidence of esophageal adenocarcinoma (EAC) has been rising dramatically in the past few decades in the United States and Western world. The N-myc downregulated gene 4 (NDRG4) belongs to the human NDRG family. In this study, we aimed to identify the expression levels, regulation, and functions of NDRG4 in EAC. Using an integrative epigenetic approach, we identified genes showing significant downregulation in EAC and displaying upregulation after 5-Aza-deoxycitidine. Among these genes, likely to be regulated by DNA methylation, NDRG4 was among the top 10 candidate genes. Analyses of TCGA (The Cancer Genome Atlas) and GEO (Gene Expression Omnibus) data sets and EAC tissue samples demonstrated that NDRG4 was significantly downregulated in EAC (p < 0.05). Using Pyrosequencing technology for quantification of DNA methylation, we detected that NDRG4 promoter methylation level was significantly higher in EAC tissue samples, as compared to normal esophagus samples (p < 0.01). A strong inverse correlation between NDRG4 methylation and its gene expression levels (r = −0.4, p < 0.01) was observed. Treatment with 5-Aza restored the NDRG4 expression, confirming that hypermethylation is a driving force for NDRG4 silencing in EAC. Pathway and gene set enrichment analyses of TCGA data suggested that NDRG4 is strongly associated with genes related to cell cycle regulation. Western blotting analysis showed significant downregulation of Cyclin D1, CDK4 and CDK6 in EAC cells after overexpression of NDRG4. Functionally, we found that the reconstitution of NDRG4 resulted in a significant reduction in tumor cell growth in two-dimensional (2D) and three-dimensional (3D) organotypic culture models and inhibited tumor cell proliferation as indicated by the EdU (5-ethynyl-2′-deoxyuridine) proliferation assay.

Porcine circovirus type 3 (PCV3) is a new member of Circovirus, which could cause various symptoms in infected pigs. It has been reported in many countries and detected from various animals. This study retrospectively analyzed serum samples that were randomly collected from 1,499 clinically healthy cattle in Shandong province from 2011 to 2018. The PCV3 DNA was detected in 28.95% (434/1,499) of samples. Twenty-seven cap genes of PCV3 were sequenced and compared with seventy reference sequences. They were in several different branches, but all belonged to PCV3b. The results indicated that PCV3 was prevalent in health cattle in Shandong province of China. Though infected cattle did not show any clinical symptoms, they could be a reservoir for the virus and probably transferred them back to pigs.

Background Pseudorabies, also known as Aujeszky's disease, is caused by the pseudorabies virus (PRV) and has been recognized as a critical disease affecting the pig industry and a wide range of animals around the world, resulting in great economic losses each year. Shandong province, one of the most vital food animal-breeding regions in China, has a very dense pig population, within which pseudorabies infections were detected in recent years. The data, however, on PRV epidemiology and coinfection rates of PRV with other major swine diseases is sparse. Objectives This study aimed to investigate the PRV epidemiology in Shandong and analyze the current control measures. Methods In this study, a total number of 16,457 serum samples and 1,638 tissue samples, which were collected from 362 intensive pig farms (≥ 300 sows/farm) covered all cities in Shandong, were tested by performing enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR). Results Overall, 52.7% and 91.5% of the serum samples were positive for PRV-gE and -gB, respectively, based on ELISA results. In addition, 15.7% of the tissue samples were PCR positive for PRV. The coinfection rates of PRV with porcine circovirus type 2 (PCV2), porcine reproductive and respiratory syndrome virus, and classical swine fever virus were measured; coinfection with PCV2 was 35.0%, higher than those of the other two viruses. Macroscopic and microscopic lesions were observed in various tissues during histopathological examination. Conclusions The results demonstrate the PRV prevalence and its coinfection rates in Shandong province and indicate that pseudorabies is endemic in pig farms in this region. This study provides epidemiological data that can be useful in the prevention and control of pseudorabies in Shandong, China.

Additional file 1: Figure S1. Library construction.

Background CD47, the integrin-related protein, plays an important role in immune resistance and escape of tumor cells. Antibodies blocking the CD47/SIRPα signal pathway can effectively stimulate macrophage-mediated phagocytosis of tumor cells, which becomes a promising approach for tumor immunotherapy. Nanobodies (Nbs) derived from camelid animals are emerging as a new force in antibody therapy. Results HuNb1-IgG4, an innovative anti-CD47 nanobody, was developed with high affinity and specificity. It effectively enhanced macrophage-mediated phagocytosis of tumor cells in vitro and showed potent anti-ovarian and anti-lymphoma activity in vivo. Importantly, HuNb1-IgG4 did not induce the agglutination of human red blood cells (RBCs) in vitro and exhibited high safety for hematopoietic system in cynomolgus monkey. In addition, HuNb1-IgG4 could be produced on a large scale in CHO-S cells with high activity and good stability. Also, we established anti-CD47/CD20 bispecific antibody (BsAb) consisted of HuNb1 and Rituximab, showing more preference binding to tumor cells and more potent anti-lymphoma activity compared to HuNb1-IgG4. Conclusions Both of HuNb1-IgG4 and anti-CD47/CD20 BsAb are potent antagonists of CD47/SIRPα pathway and promising candidates for clinical trials.

Additional file 7: Table S4. The median values of FC of each signature gene pair across different datasets for the GC, non-GC and GC adjacent-normal groups.

The incidence of esophageal adenocarcinoma (EAC) has been rising dramatically in the past few decades in the United States and Western world. The N-myc downregulated gene 4 (NDRG4) belongs to the human NDRG family. In this study, we aimed to identify the expression levels, regulation, and functions of NDRG4 in EAC. Using an integrative epigenetic approach, we identified genes showing significant downregulation in EAC and displaying upregulation after 5-Aza-deoxycitidine. Among these genes, likely to be regulated by DNA methylation, NDRG4 was among the top 10 candidate genes. Analyses of TCGA (The Cancer Genome Atlas) and GEO (Gene Expression Omnibus) data sets and EAC tissue samples demonstrated that NDRG4 was significantly downregulated in EAC (p &lt, 0.05). Using Pyrosequencing technology for quantification of DNA methylation, we detected that NDRG4 promoter methylation level was significantly higher in EAC tissue samples, as compared to normal esophagus samples (p &lt, 0.01). A strong inverse correlation between NDRG4 methylation and its gene expression levels (r = &minus, 0.4, p &lt, 0.01) was observed. Treatment with 5-Aza restored the NDRG4 expression, confirming that hypermethylation is a driving force for NDRG4 silencing in EAC. Pathway and gene set enrichment analyses of TCGA data suggested that NDRG4 is strongly associated with genes related to cell cycle regulation. Western blotting analysis showed significant downregulation of Cyclin D1, CDK4 and CDK6 in EAC cells after overexpression of NDRG4. Functionally, we found that the reconstitution of NDRG4 resulted in a significant reduction in tumor cell growth in two-dimensional (2D) and three-dimensional (3D) organotypic culture models and inhibited tumor cell proliferation as indicated by the EdU (5-ethynyl-2&prime, deoxyuridine) proliferation assay.

To explore the prognostic assessment value of preoperative blood platelet-lymphocyte ratio (PLR) in patients with gastric mixed adenoneuroendocrine carcinoma (gMANEC) treated with radical surgery.Clinical and pathological data of 84 gMANEC patients who underwent radical resection from 2006 to 2016 in Department of Gastric Surgery, Fujian Medical University Union Hospital were analyzed retrospectively. Receiver operating characteristic (ROC) curve analysis was performed to determine the cutoff value of the PLR for predicting prognosis. The Cox proportional hazards regression model was used to identify prognostic factors of gMANEC.All the patients underwent D2 lymph node dissection, including 26 cases of distal subtotal gastrectomy and 58 cases of total gastrectomy. The postoperative pathological TNM stage system(pTNM) demonstrated that the patients of stage I(, II(, and III( were 9(10.7%), 14(16.7%), and 61(72.6%) cases, respectively. The median follow-up time was 40(3 to 96) months. The recurrence rate was 41.7%(35/84). The median time to recurrence was 10 (1 to 40) months, and 82.9%(29/35) patients experienced recurrence within the first 2 years after operation. The median overall survival time was 27(3 to 39) month, and the median recurrence-free survival time was 21 (1 to 96) months. The 1-, 3-, and 5-year overall survival(OS) rates were 87.6%, 56.6%, and 47.4%, respectively, and the 1-, 3-, and 5-year recurrence-free survival (RFS) rates were 70.5%, 50.7%, and 44.9%, respectively. The best cutoff value of the PLR for predicting prognosis was 133 through ROC curve, which categorized all the patients into low PLR group (≤133) comprising 28 patients and high PLR group (133) comprising 56 patients. The tumor recurrence rate was significantly higher in high PLR group (50.0%, 28/56) than that in low PLR group(25.0%, 7/28)(P=0.028). The live metastasis rate was significantly higher in high PLR group(35.7%, 20/56) than that in low PLR group(10.7%, 3/28)(P=0.015). Cox regression analysis showed that only pTNM stage (P=0.003) was independent prognostic factors of OS, while both pTNM stage (P=0.000) and blood PLR (P=0.015) were independent prognostic factors of RFS.gMANEC patients with high preoperative PLR tend to present recurrence and metastasis, especially to present live metastasis, so they should be kept under surveillance more frequently after surgery.

To investigate the efficacy of laparoscopic surgery in the treatment of gastrointestinal stromal tumors (GIST) in different anatomical locations.Clinical data of 133 patients with primary gastric GIST undergoing laparoscopic resection at our department from January 2006 to December 2014 were retrospectively analyzed. These patients were divided into favorable site group (F group, 90 cases), including gastric fundus, anterior wall and greater curvature of gastric body, and unfavorable site group (UF group, 43 cases),including gastroesophageal junction, posterior wall and lesser curvature of gastric body,antrum and pylorus, according to the 2014 version National Comprehensive Cancer Network Clinical Guidelines. Short-term and long-term efficacy between the two groups was compared.There were no significant differences between the two groups in the general clinicopathological parameters (all P0.05). The operation time of F group and UF group was (107.3±52.3) min and (119±53.4) min respectively (P=0.21). The blood loss in F group and UF group was (35.2±34.2) ml and (35.2±31.2) ml respectively (P=1.00). In addition, there were no significant differences in time to first fluid diet, time to first flatus, postoperative hospital stay and hospitalization expenses between the two groups(all P0.05). In F group and UF group, morbidity of postoperative complication was 6.7%(6/90) and 4.7%(2/43) respectively (P=0.72), morbidity of category I(-II( complication was 4.4%(4/90) and 2.3%(1/43) respectively (P=0.66),and morbidity of category III(-IIII( complication was 2.2%(2/90) and 2.3% (1/43) respectively (P=1.00). Median follow-up time of all the cases was 36(1 to 84) months. The 5-year overall survival rates of F group and UF group were 93.8% and 95.2% respectively, and 5-year relapse-free survival rates were 81.1% and 89.4% respectively, without significant differences(both P0.05).Laparoscopic operation for gastric GIST in unfavorable sites can yield similar short- and long-term outcomes compared with those in favorable sites.

Salmonella is a zoonotic disease widely spread in the environment, causing serious economic losses to the world. Objective: To improve people's understanding of Salmonella disease and reduce economic losses. Methods: Based on the recent research progress, the biochemical characteristics, clinical manifestations, laboratory detection methods and preventive measures of bacteria were discussed in this paper. Conclusion: Salmonella infection in China is becoming more and more serious, which has caused great harm to pig industry and even human beings.

Aim: Aberrant brain functional connectivity patterns have been reported in major depressive disorder (MDD). It is unknown whether they can be used in discriminant analysis for diagnosis of MDD. In the present study we examined the efficiency of discriminant analysis of MDD by individualized computer-assisted diagnosis. Methods: Based on resting-state functional magnetic resonance imaging data, a new approach was adopted to investigate functional connectivity changes in 39 MDD patients and 37 well-matched healthy controls. By using the proposed feature selection method, we identified significant altered functional connections in patients. They were subsequently applied to our analysis as discriminant features using a support vector machine classification method. Furthermore, the relative contribution of functional connectivity was estimated. Results: After subset selection of high-dimension features, the support vector machine classifier reachedup to approximately 84% with leave-one-out training during the discrimination process. Through summarizing the classification contribution of functional connectivities, we obtained four obvious contribution modules: inferior orbitofrontal module, supramarginal gyrus module, inferior parietal lobule-posterior cingulated gyrus module and middle temporal gyrus-inferior temporal gyrus module. Conclusion: The experimental results demonstrated that the proposed method is effective in discriminating MDD patients from healthy controls. Functional connectivities might be useful as new biomarkers to assist clinicians in computer auxiliary diagnosis of MDD.

To explore the feasibility and efficacy of laparoscopic D2 radical gastrectomy in patients with gastric neuroendocrine carcinoma (GNEC).Clinical data of 84 patients with GNEC undergoing laparoscopic D2 radical gastrectomy in Union Hospital from January 2006 to December 2012 were analyzed respectively. Among these patients, 44 cases underwent laparoscopic D2 gastrectomy (LAG group) and 40 cases underwent open gastrectomy (OG group). The short- and long-term outcomes, 3-year survival and recurrence-free survival were compared between two groups.The LAG group and OG group did not differ significantly in terms of clinicopathologic characteristics. All the patients completed operations successfully and no patients in the LAG group ware converted to laparotomy. The operative time was similar (P0.05). As compared to OG group, LAG group had less intra-operative blood loss [(85±21) ml vs. (192±89) ml, P=0.003], lower ratio of transfusion [2.3%(1/44) vs. 15.0%(6/40), P=0.048], shorter time to ambulation after surgery [(2.5±1.1) days vs. (3.5±1.1) days, P=0.001], faster postoperative gastrointestinal function recovery [(2.9±1.1) days vs. (5.1±1.0) days, P=0.001], shorter time to resume soft diet [(4.1±1.2) days vs. (5.7±1.3) days, P=0.001] and shorter postoperative hospital stay [(12.0±3.4) days vs. (15.0±5.5) days, P=0.002]. No significant difference was observed in average dissected lymph node number between LAG and OG group (35.0±16.4 vs. 31.6±12.1, P=0.204). Morbidity of postoperative complication of LAG group and OG group was 11.4%(5/44) and 17.5%(7/40) respectively (P=0.422). The overall 3-year survival rate was 54.0% for all the patients, while 3-year survival rate was 56.3% in LAG group and 51.4 % in OG group (P=0.478). In addition, there was no significant difference in recurrence-free survival between the two group (33.0 months vs. 31.5 months, P=0.703).Compared with open gastrectomy, laparoscopic D2 radical gastrectomy has the advantages of faster recovery and less blood loss, and similar short-term and long-term outcomes in treatment of patients with GNEC, thus it is a safe and feasible treatment for GNEC.

Aberrant brain functional connectivity patterns have been reported in major depressive disorder (MDD). It is unknown whether they can be used in discriminant analysis for diagnosis of MDD. In the present study we examined the efficiency of discriminant analysis of MDD by individualized computer-assisted diagnosis.Based on resting-state functional magnetic resonance imaging data, a new approach was adopted to investigate functional connectivity changes in 39 MDD patients and 37 well-matched healthy controls. By using the proposed feature selection method, we identified significant altered functional connections in patients. They were subsequently applied to our analysis as discriminant features using a support vector machine classification method. Furthermore, the relative contribution of functional connectivity was estimated.After subset selection of high-dimension features, the support vector machine classifier reached up to approximately 84% with leave-one-out training during the discrimination process. Through summarizing the classification contribution of functional connectivities, we obtained four obvious contribution modules: inferior orbitofrontal module, supramarginal gyrus module, inferior parietal lobule-posterior cingulated gyrus module and middle temporal gyrus-inferior temporal gyrus module.The experimental results demonstrated that the proposed method is effective in discriminating MDD patients from healthy controls. Functional connectivities might be useful as new biomarkers to assist clinicians in computer auxiliary diagnosis of MDD.

Interleukin-1 beta (IL-1β) is one of pro-inflammatory cytokines. Recent studies have shown that IL-1β impairs hippocampal neurogenesis, mediates proliferation and differentiation of multipotent neural precursor cells (NPCs), and exerts effects of anti-proliferation, anti-neurogenesis, and pro-gliogenesis on embryonic hippocampal NPCs. The aim of this study was to examine the effect of IL-1β on the differentiation of hippocampal NPCs into functional serotonergic neurons, which play important roles in the pathophysiology and treatment of depression. Hippocampal NPCs were prepared from the hippocampus of neonatal rats (within 24h after birth). After three passages and phenotyping, hippocampal NPCs were cultured in a differentiating medium with various concentrations (5, 10, and 20 ng/mL) of IL-1β for 7 days. At the endpoint, the serotonergic differentiation of hippocampal NPCs in IL-1β-treated cultures decreased in a dose-dependent manner and this effect was blocked by IL-1ra, an IL-1 receptor antagonist capable of blocking the effects of IL-1 by binding to the same receptor (IL-1R1) without triggering signaling; serotonin in the lysate of the differentiated hippocampal NPCs decreased in IL-1β-treated cultures; and levels of Bcl-2 and phosphorylated extracellular-regulated kinase (pERK) were also lower in differentiated hippocampal NPCs with IL-1β treatment. These results support the hypothesis that IL-1β is an important factor in the stress-associated neuropathology and psychopathology and has relevance to the treatment of depressive symptoms in patients with depression.

Background Malingering detection has emerged as an important issue in clinical and forensic settings. The Structured Interview of Reported Symptoms-2 (SIRS-2) was designed to assess the feigned symptoms in both clinical and non-clinical subjects. The aim of the study was to examine the reliability and validity of the Chinese version of this scale. Methods Two studies were conducted to evaluate the reliability and validity of the Chinese Version of SIRS-2. In Study one, with a simulation design, the subjects included a. 40 students asked to simulate symptoms of mental illness; b. 40 general psychiatric inpatients and c. 40 students asked to reply to questions honestly. Scales scores for feigning symptoms among three groups were carried out for discriminant validity of the Chinese Version of SIRS-2. Minnesota Multiphasic Personality Inventory-2(MMPI-2) was administered in 80 undergraduate students. In Study two, with a known-groups comparison design, scales scores for feigning symptoms were compared between 20 suspected malingerers and 80 psychiatric outpatients from two forensic centers using the Chinese Version of SIRS-2. Results The Chinese Version of SIRS-2 demonstrated satisfactory internal consistency in both study one and two. In study one, criterion validity of this scale was supported by its significantly positive correlation with the MMPI-2 (r = 0.282 ~ 0.481 for Infrequency), and by its significantly negative correlation with the MMPI-2 (r = -0.255 ~ -0.519 for Lie and -0.205 ~ 0.391 for Correction). Scores of 10 out of 13 subscales of the Chinese Version of SIRS-2 for simulators were significantly higher than scores of honest students and general psychiatric patients. In study two, the mean scores of the Chinese Version of 13 subscales for suspected malingerers were significantly higher than those of psychiatric outpatients. For discriminant validity, it yielded a large effect size (d = 1.80) for the comparison of the participant groups in study one and two. Moreover, the sensitivity (proportion of malingerers accurately identified by the measure) and specificity (proportion of people accurately classified as responding honestly) of the Chinese version of SIRS-2 in the detection of malingering in these two studies are acceptable. Conclusions The Chinese version of the SIRS-2 has good psychometric properties and is a valid and reliable tool for detection of malingering in Chinese populations.