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Supplementary Table S1. The primers used for real-time PCR analysis Supplementary Table S2. ABPs obtained from Kyoto Encyclopedia of Genes and Genomes (KEGG) database

S1. The GEPIA website (based on TCGA data) revealed that 48 ABPs were significantly up-regulated in GC tissues. S2. H. pylori induces L-plastin expression in GC cells. S3. Direct contact is required for H. pylori induced L-plastin expression in GC cells. S4. The vacA was not involved in the induction of L-plastin. S5. Decreased expression of L-plastin in presence of U0126 during H. pylori infection. S6. L-plastin expression is increased in GC tissues.

Actin cytoskeleton dynamic rearrangement is required for tumor cell metastasis and is a key characteristic of Helicobacter pylori (H. pylori)-infected host cells. Actin cytoskeleton modulation is coordinated by multiple actin-binding proteins (ABP). Through Kyoto encyclopedia of gene and genomes database, GEPIA website, and real-time PCR data, we found that H. pylori infection significantly induced L-plastin, a key ABP, in gastric cancer cells. We further explored the regulation and function of L-plastin in H. pylori–associated gastric cancer and found that, mechanistically, H. pylori infection induced gastric cancer cells to express L-plastin via cagA-activated ERK signaling pathway to mediate SP1 binding to L-plastin promoter. Moreover, this increased L-plastin promoted gastric cancer cell proliferation and migration in vitro and facilitated the growth and metastasis of gastric cancer in vivo. Finally, we detected the expression pattern of L-plastin in gastric cancer tissues, and found that L-plastin was increased in gastric cancer tissues and that this increase of L-plastin positively correlated with cagA+ H. pylori infection status. Overall, our results elucidate a novel mechanism of L-plastin expression induced by H. pylori, and a new function of L-plastin–facilitated growth and metastasis of gastric cancer, and thereby implicating L-plastin as a potential therapeutic target against gastric cancer.Implications:Our results elucidate a novel mechanism of L-plastin expression induced by H. pylori in gastric cancer, and a new function of L-plastin–facilitated gastric cancer growth and metastasis, implicating L-plastin as a potential therapeutic target against gastric cancer.

At present, the 'green channel' policy is fully implemented. However, the detection is relatively lagging, and truck drivers are susceptible to fake the toll-free logistics vehicles (TFLVs), causing huge losses to the operation. In order to improve the accuracy and efficiency of TFLVs inspection, this paper establishes a toll evasion prediction model for fake TFLVs based on the historical TFLVs traffic dataset derived from the highway network toll collection system. First, according to the importance and reliability, we used the data mining technology to differentiate and extract the data attributes. And the spatiotemporal characteristics and other characteristics of fake TFLVs through-traffic entering and exiting toll booths are studied and analyzed based on the preprocessed data. Then use the Borderline-SMOTE oversampling method to balance the pass data set, use the ChiMerge algorithm to discretize continuous attributes. In order to ensure the effective matching and the correlation between the large contribution attributes and the results, correlation item test and collinearity test are used for discrete related attributes. Finally, adopting the processed discrete TFLVs data which passed correlation item test and collinearity test, and using the decision tree to establish the prediction model. The classification results of the escape behavior prediction model and other models are compared by using the historical TFLVs data set. The results show that the accuracy of the escape behavior prediction model proposed in this paper is 83.4%, which is higher than that of the Logistic regression model (61.8%) and the Random forest model (81%).This research model can effectively warn of fake TFLVs, and on the basis of simplifying the inspection process of TFLVs, reducing the probability of the occurrence of fake TFLVs evasion, which has practical application significance.

Actin cytoskeleton dynamic rearrangement is required for tumor cell metastasis and is a key characteristic of Helicobacter pylori (H. pylori)-infected host cells. Actin cytoskeleton modulation is coordinated by multiple actin-binding proteins (ABP). Through Kyoto encyclopedia of gene and genomes database, GEPIA website, and real-time PCR data, we found that H. pylori infection significantly induced L-plastin, a key ABP, in gastric cancer cells. We further explored the regulation and function of L-plastin in H. pylori–associated gastric cancer and found that, mechanistically, H. pylori infection induced gastric cancer cells to express L-plastin via cagA-activated ERK signaling pathway to mediate SP1 binding to L-plastin promoter. Moreover, this increased L-plastin promoted gastric cancer cell proliferation and migration in vitro and facilitated the growth and metastasis of gastric cancer in vivo. Finally, we detected the expression pattern of L-plastin in gastric cancer tissues, and found that L-plastin was increased in gastric cancer tissues and that this increase of L-plastin positively correlated with cagA+ H. pylori infection status. Overall, our results elucidate a novel mechanism of L-plastin expression induced by H. pylori, and a new function of L-plastin–facilitated growth and metastasis of gastric cancer, and thereby implicating L-plastin as a potential therapeutic target against gastric cancer. Implications: Our results elucidate a novel mechanism of L-plastin expression induced by H. pylori in gastric cancer, and a new function of L-plastin–facilitated gastric cancer growth and metastasis, implicating L-plastin as a potential therapeutic target against gastric cancer.

Background & Aims Rev-erbα represents a powerful transcriptional repressor involved in immunity. However, the regulation, function, and clinical relevance of Rev-erbα in Helicobacter pylori infection are presently unknown. Methods Rev-erbα was examined in gastric samples from H pylori-infected patients and mice. Gastric epithelial cells (GECs) were isolated and infected with H pylori for Rev-erbα regulation assays. Gastric tissues from Rev-erbα–/– and wild-type (littermate control) mice or these mice adoptively transferred with CD4+ T cells from IFN-γ–/– and wild-type mice, bone marrow chimera mice and mice with in vivo pharmacological activation or inhibition of Rev-erbα were examined for bacteria colonization. GECs, CD45+CD11c–Ly6G–CD11b+CD68– myeloid cells and CD4+ T cells were isolated, stimulated and/or cultured for Rev-erbα function assays. Results Rev-erbα was increased in gastric mucosa of H pylori-infected patients and mice. H pylori induced GECs to express Rev-erbα via the phosphorylated cagA that activated ERK signaling pathway to mediate NF-κB directly binding to Rev-erbα promoter, which resulted in increased bacteria colonization within gastric mucosa. Mechanistically, Rev-erbα in GECs not only directly suppressed Reg3b and β-defensin-1 expression, which resulted in impaired bactericidal effects against H pylori of these antibacterial proteins in vitro and in vivo; but also directly inhibited chemokine CCL21 expression, which led to decreased gastric influx of CD45+CD11c–Ly6G–CD11b+CD68– myeloid cells by CCL21-CCR7-dependent migration and, as a direct consequence, reduced bacterial clearing capacity of H pylori-specific Th1 cell response. Conclusions Overall, this study identifies a model involving Rev-erbα, which collectively ensures gastric bacterial persistence by suppressing host gene expression required for local innate and adaptive defense against H pylori., Graphical abstract

Adrenomedullin (ADM) is a multifunctional peptide that is expressed by many surface epithelial cells, but its relevance to Helicobacter pylori (H. pylori)-induced gastritis is unknown. Here, we found that gastric ADM expression was elevated in gastric mucosa of H. pylori-infected patients and mice. In H. pylori-infected human gastric mucosa, ADM expression was positively correlated with the degree of gastritis; accordingly, blockade of ADM resulted in decreased inflammation within the gastric mucosa of H. pylori-infected mice. During H. pylori infection, ADM production was promoted via PI3K–AKT signaling pathway activation by gastric epithelial cells in a cagA-dependent manner, and resulted in increased inflammation within the gastric mucosa. This inflammation was characterized by the increased IFN-γ-producing T cells, whose differentiation was induced via the phosphorylation of AKT and STAT3 by ADM derived from gastric epithelial cells. ADM also induced macrophages to produce IL-12, which promoted the IFN-γ-producing T-cell responses, thereby contributing to the development of H. pylori-associated gastritis. Accordingly, blockade of IFN-γ or knockout of IFN-γ decreased inflammation within the gastric mucosa of H. pylori-infected mice. This study identifies a novel regulatory network involving H. pylori, gastric epithelial cells, ADM, macrophages, T cells, and IFN-γ, which collectively exert a pro-inflammatory effect within the gastric microenvironment.

BHLHE40, a member of the basic helix-loop-helix transcription factor family, has been reported to play an important role in inflammatory diseases. However, the regulation and function of BHLHE40 in Helicobacter pylori (H pylori)-associated gastritis is unknown. We observed that gastric BHLHE40 was significantly elevated in patients and mice with H pylori infection. Then, we demonstrate that H pylori-infected GECs express BHLHE40 via cagA-ERK pathway. BHLHE40 translocates to cell nucleus, and then binds to cagA protein-activated p-STAT3 (Tyr705). The complex increases chemotactic factor CXCL12 expression (production). Release of CXCL12 from GECs fosters CD4+ T cell infiltration in the gastric mucosa. Our results identify the cagA-BHLHE40-CXCL12 axis that contributes to inflammatory response in gastric mucosa during H pylori infection.

Interleukin-17 receptor B (IL-17RB), a member of the IL-17 receptor family activated by IL-17B/IL-17E, has been shown to be involved in inflammatory diseases. However, the regulation and function of IL-17RB in Helicobacter pylori (H. pylori) infection, especially in the early-phase is still unknown. Here, we found that gastric IL-17RB mRNA and protein were decreased in gastric mucosa of both patients and mice infected with H. pylori. In vitro experiments show that IL-17RB expression was down regulated via PI3K/AKT pathway on gastric epithelial cells (GECs) stimulated with H. pylori in a cagA-involved manner, while in vivo studies showed that the effect was partially dependent on cagA expression. IL-17E was also decreased during the early-phase of H. pylori infection, and provision of exogenous IL-17E resulted in increased CD11b+CD11c− myeloid cells accumulation and decreased bacteria colonization within the gastric mucosa. In the early-phase of H. pylori infection, IL-17E-IL-17RB promoted gastric epithelial cell-derived CXCL1/2/5/6 to attract CD11b+CD11c− myeloid cells, and also contributed to host defense by promoting the production of antibacterial protein Reg3a. This study defines a negative regulatory network involving IL-17E, GECs, IL-17RB, CD11b+CD11c− myeloid cells, and Reg3a in the early-phase of H. pylori infection, which results in an impaired host defense within the gastric microenvironment, suggesting IL-17RB as a potential early intervening target in H. pylori infection.

Actin cytoskeleton dynamic rearrangement is required for tumor cell metastasis and is a key characteristic of

Arrestin domain containing 3 (ARRDC3) represents a newly discovered α-arrestin involved in obesity, inflammation, and cancer. Here, we demonstrate a proinflammation role of ARRDC3 in Helicobacter pylori–associated gastritis. Increased ARRDC3 was detected in gastric mucosa of patients and mice infected with H. pylori. ARRDC3 in gastric epithelial cells (GECs) was induced by H. pylori, regulated by ERK and PI3K-AKT pathways in a cagA-dependent manner. Human gastric ARRDC3 correlated with the severity of gastritis, and mouse ARRDC3 from non-BM–derived cells promoted gastric inflammation. This inflammation was characterized by the CXCR2-dependent influx of CD45+CD11b+Ly6C–Ly6G+ neutrophils, whose migration was induced via the ARRDC3-dependent production of CXCL2 by GECs. Importantly, gastric inflammation was attenuated in Arrdc3–/– mice but increased in protease-activated receptor 1–/– (Par1–/–) mice. Mechanistically, ARRDC3 in GECs directly interacted with PAR1 and negatively regulated PAR1 via ARRDC3-mediated lysosomal degradation, which abrogated the suppression of CXCL2 production and following neutrophil chemotaxis by PAR1, thereby contributing to the development of H. pylori–associated gastritis. This study identifies a regulatory network involving H. pylori, GECs, ARRDC3, PAR1, and neutrophils, which collectively exert a proinflammatory effect within the gastric microenvironment. Efforts to inhibit this ARRDC3-dependent pathway may provide valuable strategies in treating of H. pylori–associated gastritis., A regulatory network including H. pylori, GECs, ARRDC3, PAR1, and neutrophils collectively exerts a pro-inflammatory effect within the gastric microenvironment.

Cathepsin C (CtsC) functions as a central coordinator for activation of many serine proteases in immune cells. However, CtsC expression in gastric epithelial cells and its role in Helicobacter pylori infection remain unclear. Real-time PCR, Western blot, and immunohistochemistry analyses identified that CtsC was decreased in gastric mucosa of H. pylori-infected patients and mice. Isolated gastric epithelial cells and cell lines were stimulated with H. pylori and/or TGF-β1 showed that down-regulation of CtsC in gastric epithelial cells largely depended on H. pylori cagA via Src/ERK and Janus kinase (JAK)/signal transducer and activator of transcription 3 (STAT3) pathways, and the effect could be synergistically augmented by TGF-β1 in an autocrine manner. In human gastric mucosa, CtsC expression was negatively correlated with bacteria colonization; accordingly, provision of exogenous active CtsC overwhelmed H. pylori persistence in gastric mucosa of mice. In the presence of active CtsC, isolated human neutrophils activated via NF-κB pathway with augmented bactericidal capacity in vitro. We also found that neutrophils activated and cleared bacteria in active CtsC-injected mice and that there was no bactericidal capacity in mice that were simultaneously neutrophil-depleted by Ly6G antibody. Our findings identified a mechanism that H. pylori abrogate CtsC to impair neutrophil activation and to ensure persistence in gastric mucosa. Efforts to enable and boost this neutrophil activation pathway by active CtsC may therefore become valuable strategies in treating H. pylori infection.-Liu, Y. G., Teng, Y. S., Cheng, P., Kong, H., Lv, Y. P., Mao, F. Y., Wu, X. L., Hao, C. J., Chen, W., Yang, S. M., Zhang, J. Y., Peng, L. S., Wang, T. T., Han, B., Ma, Q., Zou, Q. M., Zhuang, Y. Abrogation of cathepsin C by Helicobacter pylori impairs neutrophil activation to promote gastric infection.

Background Surgical is the optimal therapeutic strategy for sacral tumors, and complete resection can effectively improve the recurrence and survival rates. However, the specialized anatomy, massive bleeding and adhesion to the anterior tissue, especially that caused by giant sacral tumors, makes complete resection difficult. The laparoscopic technique provides a new method to resect sacral tumors. Methods 34 patients with primary giant sacral tumors who underwent surgical resection were enrolled. After bilateral internal iliac artery ligation and anterior laparoscopic tumor separation, the sacral tumors were successfully resected posteriorly. The clinical, radiological and follow-up data were collected and analyzed. Results The average operative time was 276.47 min and that for laparoscopy was 76.24 min. The average intraoperative blood loss was 1757.64 ml. No complications associated with laparoscopic surgery, such as intestinal, urinary tract, or vascular injuries, occurred. Ten patients (29.41%) had perioperative complications, including infection, unhealed wounds, and cerebrospinal fluid leaks in 10, 5 and 2 patients, respectively. Patients with complications had significantly longer total (55.00 ± 34.53 vs 25.13 ± 14.60, P = 0.001) and postoperative (39.10 ± 30.61 vs 14.83 ± 10.00, P = 0.002) hospitalization stays than patients without complications. Postoperatively, bowel and bladder dysfunction, intestinal obstruction, pain, and perianal numbness occurred in 21, 5, 8, and 2 patients, respectively. The recurrence rate was 11.76%. Conclusions Laparoscopically assisted sacral tumor resection is a technically feasible and effective surgical method to resect giant sacral tumors, with the advantages of reduced operative blood loss during internal iliac artery ligation and anterior tumor separation.

Matrix metallopeptidase-10 fosters Helicobacter pylori gastric bacterial colonization and Helicobacter pylori–induced gastritis., The interaction between gastric epithelium and immune response plays key roles in H. pylori–associated pathology. We demonstrated a procolonization and proinflammation role of MMP-10 in H. pylori infection. MMP-10 is elevated in gastric mucosa and is produced by gastric epithelial cells synergistically induced by H. pylori and IL-22 via the ERK pathway. Human gastric MMP-10 was correlated with H. pylori colonization and the severity of gastritis, and mouse MMP-10 from non–BM-derived cells promoted bacteria colonization and inflammation. H. pylori colonization and inflammation were attenuated in IL-22−/−, MMP-10−/−, and IL-22−/−MMP-10−/− mice. MMP-10–associated inflammation is characterized by the influx of CD8+ T cells, whose migration is induced via MMP-10–CXCL16 axis by gastric epithelial cells. Under the influence of MMP-10, Reg3a, E-cadherin, and zonula occludens–1 proteins decrease, resulting in impaired host defense and increased H. pylori colonization. Our results suggest that MMP-10 facilitates H. pylori persistence and promotes gastritis.

Background Mast cells are prominent components of solid tumors and exhibit distinct phenotypes in different tumor microenvironments. However, the nature, regulation, function, and clinical relevance of mast cells in human gastric cancer (GC) are presently unknown. Methods Flow cytometry analyses were performed to examine level and phenotype of mast cells in samples from 114 patients with GC. Multivariate analysis of prognostic factors for overall survival was performed using the Cox proportional hazards model. Kaplan-Meier plots for patient survival were performed using the log-rank test. Mast cells, T cells and tumor cells were isolated or generated, stimulated and/or cultured for in vitro and in vivo function assays. Results Patients with GC showed a significantly higher mast cell infiltration in tumors. Mast cell levels increased with tumor progression and independently predicted reduced overall survival. These tumor-infiltrating mast cells accumulated in tumors by CXCL12-CXCR4 chemotaxis. Intratumoral mast cells expressed higher immunosuppressive molecule programmed death-ligand 1 (PD-L1), and mast cells induced by tumors strongly express PD-L1 proteins in both time-dependent and dose-dependent manners. Significant correlations were found between the levels of PD-L1+ mast cells and pro-inflammatory cytokine TNF-α in GC tumors, and tumor-derived TNF-α activated NF-κB signaling pathway to induce mast cell expression of PD-L1. The tumor-infiltrating and tumor-conditioned mast cells effectively suppressed normal T-cell immunity through PD-L1 in vitro, and tumor-conditioned mast cells contributed to the suppression of T-cell immunity and the growth of human GC tumors in vivo; the effect could be reversed by blocking PD-L1 on these mast cells. Conclusion Thus, our results illuminate novel immunosuppressive and protumorigenic roles of mast cells in GC, and also present a novel mechanism in which PD-L1 expressing mast cells link the proinflammatory response to immune tolerance in the GC tumor milieu. Electronic supplementary material The online version of this article (10.1186/s40425-019-0530-3) contains supplementary material, which is available to authorized users.

Background: Adrenomedullin (ADM) is a multifunctional peptide that is expressed by many surface epithelial cells, but its relevance to Helicobacter pylori (H. pylori)-induced gastritis is unknown. Methods: The expression of ADM was detected and localized in H. pylori infected patients and mice gastric. The signaling pathway of ADM expression was detected. The differentiation and proliferation of T cells was detected after stimulated with ADM. IL-12 was detected in macrophages after stimulated with ADM. Findings: Gastric ADM expression was elevated in gastric mucosa of H. pylori-infected patients and mice. In H. pylori-infected human gastric mucosa, ADM expression was positively correlated with the degree of gastritis, accordingly, blockade of ADM resulted in decreased inflammation within the gastric mucosa of H. pylori-infected mice. During H. pylori infection, ADM production was promoted via PI3K-AKT signaling pathway activation by gastric epithelial cells in a cagA-dependent manner, and resulted in increased inflammation within the gastric mucosa. This inflammation was characterized by the increased IFN-γ-producing T cells, whose differentiation was induced via the phosphorylation of AKT and STAT3 by ADM derived from gastric epithelial cells. ADM also induced macrophages to produce IL-12, which promoted the IFN-γ-producing T-cell responses, thereby contributing to the development of H. pylori associated gastritis. Accordingly, blockade of IFN-γ or knockout of IFN-γ decreased inflammation within the gastric mucosa of H. pylori-infected mice. Interpretation: This study identifies a novel regulatory network involving H. pylori, gastric epithelial cells, ADM, macrophages, T cells, and IFN-γ, which collectively exert a pro-inflammatory effect within the gastric microenvironment. Funding Statement: This work was funded by National Key Research and Development Program of China (grant number: 2016YFC1302200) and National Natural Science Foundation of China (grant number: 81870394). Declaration of Interests: The authors declare that they have no conflict of interest. Ethics Approval Statement: The study was approved by the Ethics Committee of XinQiao Hospital and Southwest Hospital of Third Military Medical University. All human subjects were adult, and the written informed consent was obtained from each subject. All animal experiments were performed in strict accordance with the Guide for the Care and Use of Laboratory Animals issued by the Ministry of Science and Technology of the People's Republic of China. All breeding and experiments were undertaken with review and approval from the Animal Ethical and Experimental Committee of Third Military Medical University.

Objective: To analyze the sperm quality, oxidative stress injury as well as ACP, AC and PDE expression in patients with oligoasthenozoospermia before and after qilin pill treatment. Methods: A total of 60 patients with oligoasthenozoospermia were randomly divided into observation group and control group, control group received routine western medicine treatment, observation group received qilin pill + conventional western medicine treatment, and then differences in sperm quality, oxidative stress injury, ACP, AC and PDE expression, etc. were compared between two groups after treatment. Results: Semen volume and sperm density in semen samples of observation group after qilin pill treatment were higher than those of control group; serum FSH and LH levels were lower than those of control group, and the T level was higher than that of control group; ROS and MDA levels in seminal plasma were lower than those of control group, and SOD level was higher than that of control group; ACP, AC, α-Glu and Fru levels in seminal plasma were higher than those of control group, and PDE level was lower than that of control group. Conclusion: Qilin pill can improve sperm quality and optimize testicular internal environment in patients with oligoasthenozoospermia, and it has positive clinical significance.

Objective: To study the effect of Jianpiyishen prescription on sperm quality and oxidative stress injury of testicular tissue in oligoasthenozoospermia model rats. Methods: Male SD rats were selected as experimental animals and divided into control group, model group and traditional Chinese medicine group, model group and traditional Chinese medicine group received intragastric administration of ornidazole to establish oligoasthenozoospermia models, and traditional Chinese medicine group received Jianpiyishen prescription intervention. After the intervention, sperm quality as well as ROS, SOD, GSH-Px, MDA, AC and PDE levels in testicular tissue and epididymis tissue was determined. Results: Sperm density, the number of A-grade sperm and the number of A+B-grade sperm in epididymis of model group were significantly lower than those of control group, and the sperm density, the number of A-grade sperm and the number of A+B-grade sperm in epididymis of TCM group were significantly higher than those of model group. ROS, MDA and PDE levels in testis and epididymis tissue of model group were significantly higher than those of control group, and SOD, GSH-Px and AC levels were significantly lower than those of control group; ROS, MDA and PDE levels in testis and epididymis tissue of TCM group were significantly lower than those of model group, and SOD, GSH-Px and AC levels were significantly higher than those of model group. Conclusion: After oligoasthenozoospermia model rats receive the Jianpiyishen prescription, the sperm quality is improved and the oxidative stress injury of testicular tissue is relieved.

Objective: To study the effect of ginsenoside Rh2 combined with pirarubicin on bladder cancer BIU-87 cell apoptosis and invasion. Methods: Bladder cancer BIU-87 cells were cultured and divided into control group, pirarubicin group, ginsenoside group and combined treatment group, and the cell growth and invasion as well as the expression levels of related genes was determined. Results: After treatment for 24 h, 48 h and 72 h, cell growth and invasion of pirarubicin group, ginsenoside group and combined treatment group were significantly inhibited, cell growth inhibition rate of combined treatment group was significantly higher than that of pirarubicin group and ginsenoside group, and the number of invasive cells was significantly less than those of pirarubicin group and ginsenoside group. After treatment for 24 h, 48 h and 72 h, the mRNA levels of Bcl-2 and CXCR-4 in cells and SDF-1 levels in supernatant of pirarubicin group, ginsenoside group and combined treatment group were significantly lower than those of control group, and mRNA levels of Bax in cells were significantly higher than that of control group; the mRNA levels of Bcl-2 and CXCR-4 in cells and SDF-1 level in supernatant of combined treatment group were significantly lower than those of pirarubicin group and ginsenoside group, and mRNA level of Bax in cells was significantly higher than those of pirarubicin group and ginsenoside group. Conclusion: Ginsenoside Rh2 combined with pirarubicin can promote bladder cancer cell apoptosis and inhibit bladder cancer cell invasion, and Bcl-2/Bax and SDF-1/CXCR-4 are the molecular targets of drug action.

Adrenomedullin (ADM) is a multifunctional peptide that is expressed by many surface epithelial cells, but its relevance to H. pylori-induced gastritis is unknown. Here, we found that gastric ADM expression was elevated in gastric mucosa of H. pylori-infected patients and mice. In H. pylori-infected human gastric mucosa, ADM expression was positively correlated with the degree of gastritis, accordingly, blockade of ADM resulted in decreased inflammation within the gastric mucosa of H. pylori-infected mice. During H. pylori infection, ADM production was promoted via PI3K-AKT signaling pathway activation by gastric epithelial cells in a cagA-dependent manner, and resulted in increased inflammation within the gastric mucosa. This inflammation was characterized by the increased IFN-γ-producing T cells, whose differentiation was induced via the phosphorylation of AKT and STAT3 by ADM derived from gastric epithelial cells. ADM also induced macrophages to produce IL-12, which promoted the IFN-γ-producing T-cell responses, thereby contributing to the development of H. pylori-associated gastritis. Accordingly, blockade of IFN-γ or knockout of IFN-γ decreased inflammation within the gastric mucosa of H. pylori-infected mice. This study identifies a novel regulatory network involving H. pylori, gastric epithelial cells, ADM, macrophages, T cells, and IFN-γ, which collectively exert a pro-inflammatory effect within the gastric microenvironment.Author summaryH. pylori infect almost half the world’s population. Once infected, most of people carry the bacteria lifelong if left untreated, so that persistent H. pylori infection can lead to chronic gastritis, peptic ulceration and ultimately gastric cancer. H. pylori infection is accompanied with increased inflammation in gastric mucosa, but the mechanisms of chronic gastritis induced by H. pylori infection remains poorly understood. We studied a multifunctional peptide known as adrenomedullin (ADM) in gastric epithelial cells, which was known as a key factor of regulating gastrointestinal physiology and pathology. Here, we found that gastric ADM expression was elevated in gastric mucosa of H. pylori-infected patients and mice, and was positively correlated with the degree of gastritis. ADM production was promoted via PI3K-AKT signaling pathway activation by gastric epithelial cells in a cagA-dependent manner. Blockade of ADM during H. pylori infection resulted in decreased gastric inflammation that was characterized by the increased IFN-γ-producing T cells which was induced via the phosphorylation of AKT and STAT3 by ADM derived from gastric epithelial cells. ADM also induced macrophages to produce IL-12, which promoted the IFN-γ-producing T-cell responses. These data demonstrate that H. pylori-induced ADM modulates FN-γ-producing T-cell responses and contribute to gastritis.

Heat shock proteins (HSPs) are crucial proteins in maintaining the homeostasis of human gastric epithelial cells. Tumor necrosis factor receptor-associated protein 1 (TRAP1), a member of the HSP90 family, has been shown to be involved in various crucial physiological processes, particularly against apoptosis. However, the regulation and function of TRAP1 in Helicobacter pylori infection is still unknown. Here, we found that TRAP1 expression was downregulated on human gastric epithelial cells during H. pylori infection by real-time polymerase chain reaction (PCR) and western blot analysis. Through virulence factors mutant H. pylori strains infection and inhibitors screening, we found that H. pylori vacuolating cytotoxin A ( vacA), but not cytotoxin-associated gene A ( cagA) protein, induced human gastric epithelial cells to downregulate TRAP1 via P38MAPK pathway by real-time PCR and western blot analysis. Furthermore, downregulation of TRAP1 with lentivirus carrying TRAP1 short hairpin RNA constructs impairs mitochondrial function, and increases apoptosis of gastric epithelial cells. The results indicate that H. pylori vacA downregulated TRAP1 is involved in the regulation of gastric epithelial cell apoptosis.

Interleukin-17 receptor B (IL-17RB), a member of the IL-17 receptor family activated by IL-17B/IL-17E, has been shown to be involved in inflammatory diseases. However, the regulation and function of IL-17RB in Helicobacter pylori (H. pylori) infection, especially in the early-phase is still unknown. Here, we found that gastric IL-17RB mRNA and protein were decreased in gastric mucosa of both patients and mice infected with H. pylori. In vitro experiments show that IL-17RB expression was down regulated via PI3K/AKT pathway on gastric epithelial cells (GECs) stimulated with H. pylori in a cagA-involved manner, while in vivo studies showed that the effect was partially dependent on cagA expression. IL-17E was also decreased during the early-phase of H. pylori infection, and provision of exogenous IL-17E resulted in increased CD11b+CD11c− myeloid cells accumulation and decreased bacteria colonization within the gastric mucosa. In the early-phase of H. pylori infection, IL-17E-IL-17RB promoted gastric epithelial cell-derived CXCL1/2/5/6 to attract CD11b+CD11c− myeloid cells, and also contributed to host defense by promoting the production of antibacterial protein Reg3a. This study defines a negative regulatory network involving IL-17E, GECs, IL-17RB, CD11b+CD11c− myeloid cells, and Reg3a in the early-phase of H. pylori infection, which results in an impaired host defense within the gastric microenvironment, suggesting IL-17RB as a potential early intervening target in H. pylori infection.

Myeloid-derived suppressor cells (MDSCs) are a prominent component of the pro-tumoral response. The phenotype of and mechanisms used by MDSCs is heterogeneous and requires more precise characterization in gastric cancer (GC) patients. Here, we have identified a novel subset of CD45+CD33lowCD11bdim MDSCs in the peripheral blood of GC patients compared to healthy individuals. CD45+CD33lowCD11bdim MDSCs morphologically resembled neutrophils and expressed high levels of the neutrophil marker CD66b. Circulating CD45+CD33lowCD11bdim MDSCs effectively suppressed CD8+ T cells activity through the inhibition of CD8+ T cell proliferation and interferon-γ (IFN-γ) and granzyme B (GrB) production. The proportion of CD45+CD33lowCD11bdim MDSCs also negatively correlated with the proportion of IFN-γ+CD8+ T cell in the peripheral blood of GC patients. GC patient serum-derived IL-6 and IL-8 activated and induced CD45+CD33lowCD11bdim MDSCs to express arginase I via the PI3K-AKT signaling pathway. This pathway contributed to CD8+ T cell suppression as it was partially rescued by the blockade of the IL-6/IL-8-arginase I axis. Peripheral blood CD45+CD33lowCD11bdim MDSCs, as well as IL-6, IL-8, and arginase I serum levels, positively correlated with GC progression and negatively correlated with overall patient survival. Altogether, our results highlight that a subset of neutrophilic CD45+CD33lowCD11bdim MDSCs is functionally immunosuppressive and activated via the IL-6/IL-8-arginase I axis in GC patients.

Mast cells are prominent components of solid tumors and exhibit distinct phenotypes in different tumor microenvironments. However, their precise mechanism of communication in gastric cancer remains largely unclear. Here, we found that patients with GC showed a significantly higher mast cell infiltration in tumors. Mast cell levels increased with tumor progression and independently predicted reduced overall survival. Tumor-derived adrenomedullin (ADM) induced mast cell degranulation via PI3K-AKT signaling pathway, which effectively promoted the proliferation and inhibited the apoptosis of GC cells in vitro and contributed to the growth and progression of GC tumors in vivo, and the effect could be reversed by blocking interleukin (IL)-17A production from these mast cells. Our results illuminate a novel protumorigenic role and associated mechanism of mast cells in GC, and also provide functional evidence for these mast cells to prevent, and to treat this immunopathogenesis feature of GC.

Interleukin (IL)-induced inflammatory responses are critical for the pathogenesis of Helicobacter pylori (H. pylori)-induced gastritis. IL-33 represents a recently discovered proinflammatory cytokine involved in inflammatory diseases, but its relevance to H. pylori-induced gastritis is unknown. Here, we found that gastric IL-33 mRNA and protein expression were elevated in gastric mucosa of both patients and mice infected with H. pylori, which is positively correlated with bacterial load and the degree of gastritis. IL-33 production was promoted via extracellular regulated protein kinases (ERK) signaling pathway activation by gastric epithelial cells in a cagA-dependent manner during H. pylori infection, and resulted in increased inflammation and bacteria burden within the gastric mucosa. Gastric epithelial cell-derived IL-33 promoted TNF-α production from mast cells in vitro, and IL-33 increased TNF-α production in vivo. Increased TNF-α inhibited gastric epithelial cell proliferation, conducing to the progress of H. pylori-associated gastritis and bacteria colonization. This study defined a patent regulatory networks involving H. pylori, gastric epithelial cell, IL-33, mast cell, and TNF-α, which jointly play a pathological effect within the gastric circumstances. It may be a valuable strategy to restrain this IL-33-dependent pathway in the treatment of H. pylori-associated gastritis.

Natural killer (NK) cell activity is influenced by a complex integration of signaling pathways activated downstream of both activating and inhibitory surface receptors. The tumor microenvironment can suppress NK cell activity, and there is a great clinical interest in understanding whether modulating tumor-mediated NK cell suppression and/or boosting preexisting NK cell numbers in cancer patients is therapeutically viable. To this light, we characterized the surface receptor phenotypes of peripheral blood NK cells and examined their clinical relevance to human gastric cancer (GC). We found that the proportion of peripheral blood NK cells which expressed the activating receptors NKp30, NKp46, NKG2D, and DNAM-1 was significantly decreased in GC patients compared to healthy donors, and that this decrease was positively associated with tumor progression. At the same time, plasma TGF-β1 concentrations were significantly increased in GC patients and negatively correlated with the proportion of NKp30, NKp46, NKG2D, and DNAM-1 expressing NK cells. Furthermore, TGF-β1 significantly downregulated the expression of NKp30, NKp46, NKG2D, and DNAM-1 on NK cells in vitro, and the addition of galunisertib, an inhibitor of the TGF-β receptor subunit I, reversed this downregulation. Altogether, our data suggest that the decreased expression of activating receptors NKp30, NKp46, NKG2D, and DNAM-1 on peripheral blood NK cells is positively associated with GC progression, and that TGF-β1-mediated NK cell suppression may be a therapeutically targetable characteristic of GC.

The potential contributions of CD8+ memory stem T cells to anti-tumor immunity and immunotherapy responses in gastric cancer has not been demonstrated. We found that CD8+ memory stem T cell frequencies were increased in the peripheral blood of gastric cancer patients compared to healthy donors and declined in frequency with disease progression. Despite minimal in vitro cytotoxic activity, the adoptive transfer of CD8+ memory stem T cells into Rag1−/− tumor bearing mice enhanced tumor regression compared to CD8+ central or effector memory T cell counterparts. This effect was associated with an increase in splenic, draining lymph node and tumor infiltrating CD8+ T cell numbers and the development of an altered CD8+ T cell phenotype not seen during homeostasis. GSK-3β inhibition is known to promote memory stem T cell accumulation by arresting effector T cell differentiation in vivo. Surprisingly however, GSK-3β inhibition conversely increased the cytotoxic capacity of CD8+ memory stem T cells in vitro, and this was associated with the induction of effector T cell-associated effector proteins including FasL. Finally, FasL neutralization following GSK-3β inhibition directly attenuated the anti-tumoral capacity of CD8+ memory stem T cells both in vitro and in vivo. Altogether, our findings identify the therapeutic potential of modulating CD8+ memory stem T cells for improved anti-tumoral responses against gastric cancer.

The potential contributions of CD8+ memory stem T cells to anti-tumor immunity and immunotherapy responses in gastric cancer has not been demonstrated. We found that CD8+ memory stem T cell frequencies were increased in the peripheral blood of gastric cancer patients compared to healthy donors and declined in frequency with disease progression. Despite minimal in vitro cytotoxic activity, the adoptive transfer of CD8+ memory stem T cells into Rag1−/− tumor bearing mice enhanced tumor regression compared to CD8+ central or effector memory T cell counterparts. This effect was associated with an increase in splenic, draining lymph node and tumor infiltrating CD8+ T cell numbers and the development of an altered CD8+ T cell phenotype not seen during homeostasis. GSK-3β inhibition is known to promote memory stem T cell accumulation by arresting effector T cell differentiation in vivo. Surprisingly however, GSK-3β inhibition conversely increased the cytotoxic capacity of CD8+ memory stem T cells in vitro, and this was associated with the induction of effector T cell-associated effector proteins including FasL. Finally, FasL neutralization following GSK-3β inhibition directly attenuated the anti-tumoral capacity of CD8+ memory stem T cells both in vitro and in vivo. Altogether, our findings identify the therapeutic potential of modulating CD8+ memory stem T cells for improved anti-tumoral responses against gastric cancer.

Regulatory T cells (Tregs) are major components of tumor-infiltrating immune cells with potent immunosuppressive properties in gastric cancer (GC) microenvironment. However, different subsets of the Tregs and their relevance to GC are unknown. Here, we found that patients with GC showed a significantly higher Tregs infiltration in tumors, and CD45RA−CCR7− Treg subset constituted most tumor-infiltrating Tregs. Tumor-infiltrating CD45RA−CCR7− Treg subset with an effector/memory phenotype accumulated in tumors and expressed low level of HLA-DR. Gastric tumor-derived TNF-α induced CD45RA−CCR7− Treg subset with similar phenotype to their status in tumors and inhibited their HLA-DR expression via activating STAT3 phosphorylation. These tumor-associated CD45RA−CCR7− Treg subset exerted superior immunosuppressive properties to effectively suppress CD8+ T cells’ anti-tumor function including CD8+ T-cell IFN-γ and granzyme B (GrB) production as well as CD8+ T-cell proliferation in vitro, and also contributed to the growth and progression of human gastric tumors in vivo, via IL-10 secretion and cell–cell contact mechanisms. Moreover, increased tumor-infiltrating CD45RA−CCR7− Treg subset as well as higher intratumoral CD45RA−CCR7− Treg/CD8+ T-cell ratio was associated with advanced disease progression and reduced GC patient survival. This study therefore identifies a novel immunosuppressive pathway involving CD45RA−CCR7− Treg subset development within the GC microenvironment. Efforts to inhibit this pathway may therefore prove a valuable strategy to prevent, and to treat this immune suppressive of GC.

It is presented that a new adaptive dual-voltage method to improve the control performance of high-speed solenoid valve and a control device of high-speed solenoid switch valve that could adapt to the pressure change of supply inlet is developed. Based on AMESim, this paper has processed the static and dynamic performance simulation of adaptive dual-voltage method to improve the performance of high-speed solenoid valve. This paper has built an experiment platform of adaptive dual-voltage method to improve the control performance of high-speed solenoid valve, and carried out corresponding experiments. Both simulation and experiment have proved that the method had obviously effect to improve the control performance of high-speed solenoid switch valve and the developed high-speed solenoid switch valve can effectively control the system

As the development of the Microprocessor technology, Power electronic technology, Motor manufacturing technology and modern control theory, Variable frequency system is used more and more widely.This paper introduces a design of motor variable frequency control system．AC-DC-AC-circuit is used in its main circuit, and U/F control strategy is used in its controlling method．Based on the system parameters，the calculation and selection of the component parameters are carried out．Finally，the intelligent power module(IPM)is used as a power inverter circuit switching devices and TI DSP(TMS320LF2407A) is as a core control to improve the precision and stability．The experimental result indicates that the driving system has the stable performance, high anti-interference and high value.

The alkylation reaction of guanine and N-acetylguanine with model compounds such as isopropyl bromide or 4-heptyl tosylate were studied. The reaction conditions such as temperature, solvent, base, and catalyst were examined for their effects on the reaction rate, and the yield and regioselectivity of the coupling reaction. The highest yield was obtained by using DMSO as the solvent. The reaction proceeded in a homogenous manner to give higher yield of 9-N and 7-N substituted product in a mole ratio of 1:1. The ratio could be raised to 2:1 if dibenzo-18-crown-6 was used as a catalyst. Using the above procedure, three carba-DHPG analogues bearing different 1′-C alkyl side chains were synthesized.

A new and efficient method for the synthesis of potential antiviral agents Carba-ACV (3) and Carba-DHPG (4) is described. The synthesis makes the use of N-acetylguanine to condense with the corresponding tosylates (16) and (11) under the catalysis of dibenzo-18-C-6, resulting in the desired coupling products (17) and (12) in good yield and in the case of Carba-DHPG with high regioselectivity.

Regulatory T cells (Tregs) are major components of tumor-infiltrating immune cells with potent immunosuppressive properties in gastric cancer (GC) microenvironment. However, different subsets of the Tregs and their relevance to GC are unknown. Here, we found that patients with GC showed a significantly higher Tregs infiltration in tumors, and CD45RA−CCR7− Treg subset constituted most tumor-infiltrating Tregs. Tumor-infiltrating CD45RA−CCR7− Treg subset with an effector/memory phenotype accumulated in tumors and expressed low level of HLA-DR. Gastric tumor-derived TNF-α induced CD45RA−CCR7− Treg subset with similar phenotype to their status in tumors and inhibited their HLA-DR expression via activating STAT3 phosphorylation. These tumor-associated CD45RA−CCR7− Treg subset exerted superior immunosuppressive properties to effectively suppress CD8+ T cells’ anti-tumor function including CD8+ T-cell IFN-γ and granzyme B (GrB) production as well as CD8+ T-cell proliferation in vitro, and also contributed to the growth and progression of human gastric tumors in vivo, via IL-10 secretion and cell–cell contact mechanisms. Moreover, increased tumor-infiltrating CD45RA−CCR7− Treg subset as well as higher intratumoral CD45RA−CCR7− Treg/CD8+ T-cell ratio was associated with advanced disease progression and reduced GC patient survival. This study therefore identifies a novel immunosuppressive pathway involving CD45RA−CCR7− Treg subset development within the GC microenvironment. Efforts to inhibit this pathway may therefore prove a valuable strategy to prevent, and to treat this immune suppressive of GC.

For the spacecraft assembly Process Bill of Material (PBOM) traditional construction technology, the product assembly relationship, product assembly information are not visual for spacecraft assembly and integration department, leading to the manual PBOM construction by using Product Data Management (PDM) system, which is inefficient and unreliable. This paper sorts all spacecraft products into seven Primary Product and others are Affiliated Product. Then this paper proposes the rules of PBOM automatic construction technology, which are Product Types mapping rule, assembly constraint rule and geometric rule, and introduces the PBOM automatic construction principle and workflow based on Pro/E. Finally, according to a real satellite manufacture, this paper describes the improved efficiency and reliability of the software developed with the technology, realizes the import into the PDM system, and analyzes the fault of the technology.

It is uncertain whether gender affects the outcomes of catheter ablation (CA) for atrial fibrillation (AF). The objective of the study is to evaluate the efficacy and safety of CA for long-standing persistent AF in women.Between January 2010 and May 2011, 220 consecutive patients (73 females, 33.2%), with long-standing persistent AF who underwent CA were prospectively recruited. Gender-related differences in clinical presentation, periprocedural complications, and outcomes were compared.Women were less likely to have lone AF than men (27.4% vs 47.6%; P = 0.004). The incidence of rheumatic heart disease was higher in women (19.2% in women vs 1.4% in men; P0.001). Women had a lower initial ablation success rate than men (35.6% vs 57.1%; P = 0.003). Hematomas occurred more often in women (6.8% in women vs 0.7% in men; P = 0.027). A Cox regression analysis demonstrated total duration of AF (per month, hazard ratio [HR] 1.003, confidence interval [CI] 1.001-1.006; P = 0.006) and gender (HR 1.663, CI 1.114-2.485; P = 0.013) as the independent predictors for recurrence after the first CA.Women and long AF duration were closely related to the recurrence of AF after the first ablation in patients with long-standing persistent AF. Women also had a higher risk of vascular complications.

This randomized prospective study compared three ablation strategies in patients with long-standing persistent atrial fibrillation (LPeAF). It also explored the best procedural endpoint from among the following: circumferential pulmonary vein isolation (PVI) + left atrial (LA) linear lesions (roofline, mitral isthmus) + complex fractionated atrial electrogram (CFAE) ablation, PVI + LA linear lesions + cavotricuspid isthmus (CTI) ablation + CFAE ablation, and PVI + CFAE ablation.A total of 210 patients with LPeAF referred for catheter ablation were enrolled and randomized into three ablation groups. The patients in group A (n = 70) underwent PVI followed by LA linear and CFAE ablation; in 93% of patients the primary endpoint was achieved (five patients with incomplete linear lesions). Of the 70 patients in group B who were subjected to PVI followed by LA linear, CFAE, and CTI ablations, in 94% of patients the primary endpoint was achieved (four patients with incomplete linear lesions). All patients in group C (n = 70) successfully underwent PVI and CFAE ablation. Direct current cardioversion was performed upon PVI, CFAE elimination, and completion of linear lesions. Patients were followed-up for atrial tachyarrhythmia recurrence for at least 24 months. After a single ablation procedure, group C (36%) exhibited the lowest success compared with group A (54%) and group B (51%) (P = 0.06). At the mean follow-up of 32 ± 9 months after the final ablation procedure, 53 patients (76%) in group A, 53 (76%) in group B, and 41 (59%) in group C were in sinus rhythm without antiarrhythmic drugs (P = 0.03).In LPeAF, linear lesions in the LA help improve outcome of ablation, additional CTI ablation does not.

Our objective was to observe the role of vascular endothelial growth factor (VEGF) 121 gene transfer in promoting vascular reconstruction and bone repair in femur head necrosis of rabbits.The femoral head necrosis model was induced by injection with ethanol. The necrotic femoral head was transfected with a human adenoviral vector expressing VEGF (Ad-hVEGF121). Bone formation in the subchondral necrotic region was analyzed using histology, by measuring the bone mineral density value, and by observing bone trabecular morphology using image analysis.Revascularization level, bone formation rate, bone quality and quantity, and mineralization level in the subchondral necrotic region of the gene transfection group were significantly higher than the control groups. The control groups had more subchondral bone resorption compared with the gene transfection group.VEGF might promote bone formation and revascularization in the subchondral necrotic region of the femoral head, indirectly protecting the necrotic bone trabecula from absorption and avoiding a reduction in the mechanical function of the subchondral region.

To approach the expression and its significance of connection tissue growth factor in articular cartilage during repair after full-thickness cartilage injury in young rabbit model.Young rabbit model of spontaneous healing of full-thickness cartilage defect in the medial femoral knee condyle was reproduced in present study. A total of twenty-five young New Zealand white rabbit were randomly divided into five groups: control group, 24 hour group, 1 week group, 4 week group and 8 week group after full-thickness cartilage injury (5 each). The mRNA and protein expression of CTGF were detected with RT-PCR and immunohistochemistry, respectively.The morphologically good repairs with hyaline-like cartilage appearance were observed at 8 weeks after injury. The expression of CTGF mRNA was detected in all groups. Expression of CTGF mRNA was significantly higher in all groups after cartilage injury than in control group(P0.05).The immunohistochemistry results showed that the expression of CTGF was positive in all groups, and immunolocalization was observed in the middle and deep zone.The up-regulated expression of CTGF is accompanied with spontaneous healing of full-thickness cartilage defect in young rabbit, and CTGF may play a role in the repair of articular cartilage injury.

Objective: To study the role of vascular endothelial growth factor (VEGF) in the process of fracture healing and the effect of VEGF and anti-VEGF polyclonal antibody on fracture healing. Methods: One hundred and five New Zealand white rabbits were subjected to fracture of the middle part of the left radius, and were randomly divided into control, VEGF, and VEGF polyclonal antibody groups. The blood flow at the fracture site was measured by single photoemission computerized tomography after 8 hours, 24 hours, and 72 hours, and 1 weeks, 3 weeks, 5 weeks, and 8 weeks. X-ray films were taken after 1 weeks, 3 weeks, 5 weeks, and 8 weeks to observe the results of fracture healing. Results: The blood flow at the fracture site in the VEGF group significantly increased compared with the control group during 8 hours to 1 week, but no obvious difference was seen on the X-ray films between the two groups. In the VEGF polyclonal antibody group, the blood flow at the fracture sites decreased significantly at all time points compared with the control group. The fracture healing process was disturbed, and nonunion signs were seen at the fracture site. Conclusions: The lack of VEGF may impede the fracture healing process, and results in nonunion at the fracture site.

To investigate the pathological expression and significance of VEGF in patients with active ankylosing spondylitis.The expression of VEGF in the synovial tissues of cacroiliac joint of patients with active AS was detected by using in situ hybridization and the results were compared with those in the patients with pelvic fracture using image analysis system.The positive expressions of VEGF in the synovial tissues of cacroiliac joint of patients with active AS were stronger than those in the control group (P0.01).VEGF are important factors in patients with active AS. They are tightly correlated with the process of osteoclasia and pathological new bone formation in the cacroiliac joint of patients with active AS. If we can reduce the expressions of VEGF in the patients with active AS, the process of osteoclasia and pathological new bone formation will be interrupted and this provides a new strategy for the treatment of ankylosing spondylitis.

This study investigated changes in expression of endothelial-overexpressed lipopolysaccharide-associated factor 1 (EOLA1) in grafts following liver transplantation in rats. Thirty Lewis rats received liver transplants from Lewis rats (tolerance group); 30 received liver transplants from dark Agouti rats (acute rejection group). Changes in serum biochemical indexes (alanine aminotransferase and total bilirubin), graft histology and EOLA1 expression were measured on days 1, 3, 5, 7 and 10 post-operatively. Mean survival time was >100 days in the tolerance group and 16.2 ± 1.4 days in the acute rejection group. Pathological evidence of acute rejection in grafts was seen after day 5 in the acute rejection group. Serum biochemical indexes were significantly higher in the acute rejection group than in the tolerance group from day 5 post-operatively, whereas EOLA1 expression in the liver graft was significantly higher in the tolerance group than in the acute rejection group. EOLA1 expression seems to be negatively correlated with severity of rejection after liver transplantation.

To clone and identify one novel regeneration related gene H(3) (CA854305) from the differential expression genes library we had set up before.Use the method of Northern blot to detect the different expressions of the novel gene under different situations, employ the technique of in silico cloning to scan the span of the novel gene, and analyze their sequences. Also we used reverse transcription PCR to validate the largest open reading frame.Northern blotting results of H(3) (CA854305) showed that the transplanted group had more efficient and extensive expression than untreated and uninjured groups 5 days after spinal cord injury, while the untreated group had more extensive expression than uninjured group. It implied that H(3) might have some relationship with nerve regeneration after spinal cord injury. From the results of in silico cloning we got a longest contig of 1635 bp and an largest open reading frame of 542 bp from 49 to 591 bp correspondent with the Cozak rules. Reverse transcription PCR validated the largest open reading frame sequence primarily.We got the sequence of novel gene H(3) which might be one of the regenerationjrelated genes. Key words:Gene library; Genes; Nerve regeneration; Spinal cord injuries.