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Additional file 5: Figure S4II. Interaction between CLK1 and SRSF5 in PANC-1 cells, BxPC-3 cells, and 293T cells. (A-E) HA-tagged CLK1 and Flag-tagged SRSF5 were transfected into PANC-1 cells (A), or BxPC-3 cells (B-C), or 293T cells (D-3). Co-immunoprecipitation and western blot assays were performed with the indicated antibodies.

Additional file 1: Figure S1. Higher CLK1 expression in tumors correlated with worse prognosis of PDAC patients. (A) The expression of CLK1 in pancreatic tumor and adjacent normal tissues from TCGA database were analyzed. (T = 179, N = 171). (B) The expression of CLK1 in 156 paraffin-embedded specimens from the external cohort was determined by TMA-based IHC staining. Representative IHC images are shown. (C)The relative CLK1 staining intensity was scored. (D–H) Kaplan–Meier analyses of the correlations between CLK1 expression and overall survival (D), disease-free survival (E) of all PDAC patients, or Stage II patients (F, I), or Stage III patients (G, H) in the internal cohort. *P

Additional file 4: Figure S4I. CLK1 and SRSF5 had prognostic values for PDAC patients. (A) The quantification of the expression level of p-SRSF5 in different PC cell lines. (B) The quantification of the results of Fig. 4L. (C) The quantification of the results of Fig. 4M. (D, E) The phosphorylation level of SRSF5 on Ser250 rely on the expression of CLK1. (B-J) Kaplan–Meier analyses of the correlations between CLK1/SRSF5 expression and overall survival or disease-free survival of PDAC patients.

Additional file 7: Figure S7. METTL14 exon skipping functioned downstream of the CLK1 signaling to control the malignant behaviors of pancreatic cancer cells. (A-D) PANC-1 cells were infected with control lentivirus, or lentivirus expressing CLK-1 alone or CLK-1 together with shRNA specific to METTL14exon10+. The m6A level (A), colony-formation ability (B), cell proliferation ability (C), and migration and invasion ability (D) of the indicated stable cells were evaluated. (E–H) PANC-1 cells were infected with control lentivirus, or lentivirus expressing CLK-1-specific shRNA or CLK-1-specific shRNA together with METTL14exon10+. The m6A level (E), colony-formation ability (F), cell proliferation ability (G), and migration and invasion ability (H) of the indicated stable cells were evaluated. (I-L) PANC-1 cells were treated with TG003 alone or together lentivirus infection-mediated overexpression of METTL14-L or METTL14-S. The m6A level (I), colony-formation ability (J), cell proliferation ability (K), and migration and invasion ability (L) of the indicated stable cells were evaluated. n = 3 for each group; data are shown as mean ± SD for three independent experiments. *P

Additional file 3: Figure S3. The impacts of CLK1 overexpression and knockdown on the migration and invasion of 8988 cells. (A-H) 8988 cells with stable CLK1 overexpression (A-D) and with CLK1 knockdown (E–H) were generated. Cell migration and invasion ability (A-C, E–G), and wound healing ability (D, H) in the indicated cell lines were evaluated. n = 3 for each group; data are shown as mean ± SD from three independent experiments. *P

Additional file 9: Figure S9. Aberrant alternative splicing of METTL14△exon10 and CyclinL2△exon6.3 possessed prognostic values for PDAC patients in external corhort.

Additional file 2: Figure S2. The impacts of CLK1 overexpression and knockdown on the proliferation of 8988 cells. (A, F) 8988 cells with stable CLK1 overexpression or Knockdown were confirmed by western blot. Cell proliferation (B, C, G, H), colony formation (D, I), and cell cycle progression (E, J) in the indicated cell lines were evaluated. n = 3 for each group; (K) The expression of CLK1, p53 and p21 in the tissues from the xenograft tumor were determined by TMA-based IHC staining. Representative IHC images are shown. (L-M) The expressions of p53 and p21 were negatively correlated with the expression of CLK1 in the xenograft tumors. Data are shown as mean ± SD from three independent experiments. *P

Additional file 8: Figure S8. CyclinL2△exon6.3 skipping functioned downstream of the CLK1 signaling to control the malignant behaviors of pancreatic cancer cells. (A-C) PANC-1 cells were infected with control lentivirus, or lentivirus expressing shRNA-specific to SRSF5 alone or together with overexpression of CCNL2-L or CCNL2-S. The cell proliferation ability (A), colony-formation ability (B), and migration and invasion ability (C) of the indicated stable cells were evaluated. (D-F) PANC-1 cells were infected with control lentivirus, or lentivirus expressing shRNA-specific to CLK1 alone or together with overexpression of CCNL2-L or CCNL2-S. The cell proliferation ability (D), colony-formation ability (E), and migration and invasion ability (F) of the indicated stable cells were evaluated. (G-I) PANC-1 cells were infected with control lentivirus, or lentivirus expressing CLK1 alone or CLK1 together with lentivirus expressing shRNA-specific to CCNL2-L. The cell proliferation ability (G), colony-formation ability (H), and migration and invasion ability (I) of the indicated stable cells were evaluated. (J-L) PANC-1 cells were treated with TG003 alone or together with lentivirus infection-mediated overexpression of CCNL2-L or CCNL2-S. The cell proliferation ability (J), colony-formation ability (K), and migration and invasion ability (L) of the indicated stable cells were evaluated. (M) The quantification of the results of Fig. 8n, o. n = 3 for each group; data are shown as mean ± SD from three independent experiments. *P

Additional file 6: Figure S5. CLK1-mediated SRSF5 phosphorylation on Ser250 contributed to the malignant behaviors of pancreatic cancer cells. (A-E) BxPC-3 cells were infected with control lentivirus, or lentivirus expressing wild type (WT)/mutated (S250MU)/phosphorylated (S250P) SRSF5. The cell proliferation ability (A, B), colony formation ability (C), and migration and invasion ability (D, E) of the indicated stable cells were evaluated. (F-J) PANC-1 cells were infected with control lentivirus or lentivirus overexpressing CLK1 or lentivirus overexpressing CLK1 together with shSRSF5. The cell proliferation ability (F, G), colony formation ability (H), and migration and invasion ability (I, J).The stable cells were infected with control lentivirus, or lentivirus expressing wild type (WT)/mutated (S250MU)/phosphorylated (S250P) SRSF5 were simultaneous treatment with TG003 (K–O)were evaluated. The cell proliferation ability (K, L), colony formation ability (M), and migration and invasion ability (N, O). (P)The quantification of the results of Fig. 5B. (Q, R)The quantification of the results of Fig. 5K. n = 3 for each group; data are shown as mean ± SD from three independent experiments.*P

Background: Pancreatic cancer(PDAC) remains one of the most lethal cancers worldwide, The accumulation of many abnormal epigenetic abnormalities contribute to the fatal prognosis of PDAC,While available studies are still limited. Many studies have confirmed that the Shc SH2-domain binding protein 1(SHCBP1) plays as proto-oncogene in cancers. However, whether SHCBP1 plays a role in pancreatic cancer oncogenesis is still unknown.EGF domain-specific O-linked GlcNActransferase (EOGT) acts as a key participant in the O-GlcNAcylation of NOTCH1. EOGT mutations inhibit the NOTCH1 signalling pathway and cause several congenital developmental disorders, However, the role of EOGT in malignancies has not been reported.Methods:SHCBP1 and EOGT were identified as proto-oncogenes in PDAC by High-throughput sequencing and Cox regression analysis,and validated by Internal and External cohort and public databases. The function of SHCBP1 and EOGT was determined in vitro and in vivo, The underlying mechanism was investigated by Western blot,Immunofluorescence（IF），Co-immunoprecipitation (Co-IP), Chromatin immunoprecipitation (ChIP)，and luciferase analyses.Results: The expression of EOGT and SHCBP1 was significantly elevated and correlated with worse prognosis in PDAC patients. In vitro, SHCBP1 overexpression promoted pancreatic cancer cell proliferation,migration and invasion, while knocking down SHCBP1 and EOGT inhibited these malignant processes. In vivo data showed that SHCBP1 overexpression promoted xenograft growth and lung metastasis and shortened survival in mice,whereas knocking down either EOGT or SHCBP1 expression suppressed xenograft growth and metastasis and prolonged survival. Morever，EOGT and SHCBP1 enhance the O-GlcNAcylation of NOTCH1, subsequently promoting the nuclear localization of the Notch intracellular domain (NICD) and inhibiting the transcription of E-cadherin and P21 in pancreatic cancer cells.Conclusion:All in all，This research revealed that SHCBP1 and EOGT act as proto-oncogenes in PDAC by enhancing the O-GlcNAcylation of NOTCH1，Which provides a new field of vision for new therapeutic targets for pancreatic cancer.

Serine/arginine-rich splicing factor 3 (SRSF3) regulates mRNA alternative splicing of more than 90% of protein-coding genes, providing an essential source for biological versatility. This study finds that SRSF3 expression is associated with drug resistance and poor prognosis in pancreatic cancer. We also find that SRSF3 regulates ANRIL splicing and m6A modification of ANRIL in pancreatic cancer cells. More importantly, we demonstrate that m6A methylation on lncRNA ANRIL is essential for the splicing. Moreover, our results show that SRSF3 promotes gemcitabine resistance by regulating ANRIL's splicing and ANRIL-208 (one of the ANRIL spliceosomes) can enhance DNA homologous recombination repair (HR) capacity by forming a complex with Ring1b and EZH2. In conclusion, this study establishes a link between SRSF3, m6A modification, lncRNA splicing, and DNA HR in pancreatic cancer and demonstrates that abnormal alternative splicing and m6A modification are closely related to chemotherapy resistance in pancreatic cancer.

Yuqiao Reservoir, an important adjusting reservoir in the project of diverting water from the Luanhe River to Tianjin City, is an important water resource for Tianjin City. Concentrations and spatial variation characteristics of Cd, As, Pb, Cu, Cr, and Zn from 33 sub watershed sampling sites of Yuqiao reservoir and the relationship to landscape pattern were discussed. The result showed that average concentrations of Cd, As, Cu, Cr in suspended particulate matter were higher than Chinese environmental quality standard Grade one for soils, and there were ecological risks for them because they could arrive at Yuqiao reservoir. The average concentrations of Cd, As, Pb, Cu, Cr, and Zn in surface sediments were 0.32 mg·kg

Based on the study of abroad foundation on visual theory, we conduct depth discussion of the current introduction of visual teaching meaning. For the current computer graphics course content, we determine a visual teaching content and objectives, carry out the study and practice of computer graphics algorithm visualization teaching. This paper uses VB 6.0 visual programming tool to generate the basic primitives, polygon filling, cut a straight line segment, curve generation and visualization of fractal algorithm design and realization. Through analysis, this paper hold that use of visual teaching has a positive effect and influence for professors to change the way teachers help students understand computer graphics algorithm. In the process of teaching, students improve the understanding, exploration, discovery and creativity through observation, conjecture and verify.

A new montmorillonite/humic acid complex preparation method in alkaline environment was studied by experiment, and the complex effect on remediation of heavy metal pollution was verified. The best technologic condition of the montmorillonite/humic acid complex preparation in alkaline environment was that the mass ratio between montmorillonite and humic acid was 100:3, the reaction solution was 0.01 mol/L sodium nitrate solution, the pH value of the reaction solution was 8.5, and the reaction time was 24 hours. The complex has better remediation effect on cadmium waste water. The cadmium adsorption capacity of the complex was 18.96 mg/g, and the pH value ranges of cadmium waste water suited for the complex was from 5 to 9. When the ratio of Cd initiative concentration/the complex quantity was lower than 300:1 and the pH value of the waste water was higher than 8, almost all of the Cd in waste water can be removed.

The impact of inorganic anions on cadmium (Cd) fractions in soils was studied by experiment. The result showed that Cd exchangeable fraction contents in soils increased with the increase in soil salinity during soil saline process; Cd exchangeable fraction contents in soils increased with the increase Cl- concentration of soil solutions, other fractions of Cd decreased with the increase of of Cl- concentration; the changes of SO42- concentration in soil solutions did not affect on Cd exchangeable fraction contents in soils, Cd carbonate fraction of increased, but the concentrations of Cd oxFe-Mn fraction and organic bound fraction decreased with the increase of SO42- concentration; Cd exchangeable fraction content reduced significantly with the increase in HCO3- concentration of soil solutions, Cd carbonate fraction increased, and the concentrations of Cd oxFe-Mn fraction and organic bound fraction changed insignificant.

CDK5RAP3 was isolated as a binding protein of the Cdk5 activator p35. Although CDK5RAP3 has been implicated in cancer progression, its expression and function have not been investigated in gastric cancer. Our study demonstrated that the mRNA and protein levels of CDK5RAP3 were markedly decreased in gastric tumor tissues when compared with respective adjacent non-tumor tissues. CDK5RAP3 in gastric cancer cells significantly reduced cell proliferation, migration, invasion and tumor xenograft growth through inhibition of β-catenin. Secondly, CDK5RAP3 was found to suppress the phosphorylation of GSK-3β (Ser9), leading to the phosphorylation (Ser37/Thr41) and subsequent degradation of β-catenin. Lastly, the prognostic value of CDK5RAP3 for overall survival was found to be dependent on β-catenin cytoplasm/nucleus localization in human gastric cancer samples. Collectively, our results demonstrated that CDK5RAP3 negatively regulates the β-catenin signaling pathway by repressing GSK-3β phosphorylation and could be a potential therapeutic target for gastric cancer.

In order to use the Cd-contaminated saline soil, experiments have been carried out to analyze the differences among effects of three salts on cadmium accumulation capacities of Brassica Napus in Cd-contaminated soil, thus to figure out the phytoremediation effects of planting Brassica Napus in different types of Cd-contaminated saline soils. Brassica Napus(a cadmium hyperaccumulator plant) has been as the research plant, the Brassica Napus was planted in Cd-contaminated soils (Cd: 10 mg•kg-1) with different salt concentrations(0 g•kg-1, 2g•kg-1,4 g•kg-1 and 6 g•kg-1) for 60 days as required by the greenhouse pot soil culture experiment, thus to study the bioconcentration factor(BCF) of Brassica Napus on Cd and the effects of Brassica Napus on the changes of concentrations in the shoots and roots. The three main salts in the soil, namely, sodium chloride, sodium sulfate and sodium carbonate, were chosen as the analysis and research objects. The results showed that the soil containing sodium carbonate inhibited the Brassica Napus from absorbing Cd in the soil, so did the soil containing sodium sulfate, however, the effect was not so obvious as that of the soil containing sodium carbonate. However, the soil containing sodium chloride had little impact on Cd absorption of the Brassica Napus that it could only slightly promote the cadmium accumulation capacities of Brassica Napus under a very high concentration, In different types of saline soils, there were significant differences among the effects of different salts on cadmium accumulation capacities of Brassica Napus, the sodium chloride in the soil had little impact on cadmium accumulation capacities of the roots of Brassica Napus, however, it could enhance the cadmium accumulation capacities of the shoots of Brassica Napus; the sodium carbonate in the soil could significantly inhibit the shoots and roots of Brassica Napus from accumulating the cadmium, therefore, it was not conducive for the Brassica Napus to accumulate cadmium.

A new cell line was established from a liver metastasis of a human pancreatic adenocarcinoma. The cell line, MDAPanc-3, which arose from a moderately differentiated adenocarcinoma, produces carbonic anhydrase II mRNA, but no detectable levels of insulin or alpha amylase mRNA. The stem line chromosome number was determined to be 43, with six marker chromosomes. Growth of MDAPanc-3 is stimulated by cholecystokinin (CCK) fragment 26-33. The cell line will be useful in further studies on the mechanism(s) by which CCK stimulates growth of certain human pancreatic adenocarcinomas and normal human pancreatic exocrine tissue.