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Long non-coding RNAs modulate tumor occurrence through different molecular mechanisms. It had been reported that HNF1A-AS1 (HNF1A Antisense RNA 1) was differently expressed in multiple tumors. The role of HNF1A-AS1 in colorectal cancer was less analyzed, and the mechanism of regulating the cell cycle has not been completely elucidated.Differentially expressed lncRNAs were screened out from the TCGA database. HNF1A-AS1 was examined in CRC clinical samples and cell lines by RT-qPCR. CCK8 assay, colony formation assay, flow cytometry, transwell assays, tube forming assay and vivo experiments were performed to study the function of HNF1A-AS1 in CRC tumor progression. Bioinformatic analysis, luciferase report assay, RNA pull-down and RIP assays were carried out to explore proteins binding HNF1A-AS1 and the potential downstream targets.Our results showed that HNF1A-AS1 was upregulated in CRC and associated with unfavorable prognosis. HNF1A-AS1 promoted proliferation, migration and angiogenesis, accelerated cell cycle and reduced cell apoptosis in CRC. Bioinformatics prediction and further experiments proved that HNF1A-AS1 could promote CCND1 expression by suppressing PDCD4 or competitively sponging miR-93-5p. Meanwhile, METTL3 mediated HNF1A-AS1 mIn summary, our result reveals the novel mechanism in which m

Several photosensitizers have recently been proposed as novel approaches against β-lactamase-producing drug-resistant bacteria. However, these reported photosensitizers are rarely used for accurate recognition of drug-resistant bacteria. To tackle this challenge, the structurally modified photosensitizer CySG-2 based on a lipophilic cationic heptamethine indocyanine near-infrared (NIR) dye (IR-780) and an important synthesis intermediate of cephalosporin antibiotic (GCLE) not only achieved the accurate recognition of TEM-1 methicillin-resistant

Introduction LncRNA MIR503HG has been reported to participate in liver cancer and ALK-negative anaplastic large-cell lymphoma, while its role in non-small cell lung cancer (NSCLC) is unknown. We therefore investigated the functions of lncRNA MIR503HG in NSCLC. Methods MIR503HG expression in paired cancer and non-cancer tissues from NSCLC patients was analyzed by RT-qPCR. The interaction between cyclin D1 and MIR503HG was analyzed by overexpression experiments. Cell cycle analysis was performed by flow cytometry. Cell proliferation was analyzed by CCK-8 assay. Results MIR503HG was downregulated in NSCLC and low levels of MIR503HG were associated with poor survival. In contrast, cyclin D1 was upregulated in NSCLC, and cyclin D1 and MIR503HG were inversely correlated. In NSCLC cells, overexpression experiments revealed that MIR503HG functioned as an upstream inhibitor of cyclin D1. MIR503HG overexpression led to G1 cell cycle arrest, while overexpression of cyclin D1 attenuated the effects of MIR503HG overexpression. Similarly, MIR503HG overexpression resulted in reduced cell proliferation rate, while overexpression of cyclin D1 caused the increased cell proliferation rate and attenuated effects of MIR503HG overexpression. Conclusion MIR503HG inhibits NSCLC cell proliferation by inducing cell cycle arrest through the downregulation of cyclin D1.

Nitric oxide (NO) has an important class of endogenous diatomic molecules that play a key regulatory role in many physiological and biochemical processes. However, the type of nitrosamine NO donor stimulated by light has many advantages compared to the conventional NO donors such as diazeniumdiolates and

This study aimed to evaluate the role of human epididymis protein 4 (HE4) in the diagnosis and determination of the severity of interstitial lung disease (ILD) in rheumatoid arthritis (RA) patients.HE4 levels in peripheral blood (PB) and bronchoalveolar lavage fluid (BALF) samples were determined via electrochemiluminescence immunoassays in 102 RA patients (46 patients with ILD and 56 patients without ILD) and 51 healthy controls (HCs).Serum HE4 levels were significantly higher in RA-ILD patients (141.8±65.92 pmol/l) than those in the RA-no ILD patients (82.67±26.17 pmol/l) and healthy controls (35.72±7.6 pmol/l) (p0.0001). Consistent with serum HE4 levels, BALF HE4 levels were significantly higher in RA-ILD patients (637.6±154.9 pmol/l) than those in the RA-no ILD patients (427.3±111.2 pmol/l) and healthy controls (206.9±30.46 pmol/l) (p0.0001). In RA-ILD patients, HE4 levels were positively correlated with HRCT (high-resolution computed tomography) fibrosis scores, whereas a significant inverse relationship was found between HE4 levels and lung function parameters (such as, diffusion capacity of the lung for carbon monoxide (DLCO)). The logistic regression analysis showed that high levels of BALF HE4 (≥595 pmol/l) were associated with RA-ILD (odds ratio [OR] =8.09; 95% confidence interval [CI] =1.317-49.682; p=0.024).Serum and BALF HE4 levels were elevated in RA-ILD patients and strongly associated with the severity of ILD, thus supporting their potential clinical value as a new diagnostic aid for patients with RA-ILD.

ObjectiveTo test the antifibrotic effect of dermatan sulphate in a bleomycin-induced mouse model of pulmonary fibrosis.MethodsC57 mice were randomly divided into four experimental groups: saline-treated control group, bleomycin-induced fibrosis group, prednisolone acetate group and dermatan sulphate group. Lungs were assessed using the lung index, and the extent of interstitial fibrosis was graded using histopathological observation of haematoxylin & eosin-stained lung tissue. Lung tissue hydroxyproline levels and blood fibrinogen levels were measured using a hydroxyproline colorimetric kit and the Clauss fibrinogen assay, respectively. Tissue-type plasminogen activator (tPA) was measured using a chromogenic tPA assay kit.ResultsLung index values were significantly lower in the dermatan sulphate group versus the fibrosis group. Histopathological analyses revealed that dermatan sulphate treatment ameliorated the increased inflammatory cell infiltration, and attenuated the reduction in interstitial thickening, associated with bleomycin-induced fibrosis. Hydroxyproline and fibrinogen levels were decreased in the dermatan sulphate group versus the fibrosis model group. Dermatan sulphate treatment was associated with increased tPA levels versus controls and the fibrosis group.ConclusionsDamage associated with bleomycin-induced pulmonary fibrosis was alleviated by dermatan sulphate.

Background Long noncoding RNAs (lncRNAs) are involved in various human diseases, including cancers. However, their mechanisms remain undocumented. We investigated alterations in lncRNA that may be related to pancreatic cancer (PC) through analysis of microarray data. Methods In the present study, quantitative real-time PCR analysis was used to examine the expression of taurine upregulated 1 (TUG1) in PC tissue samples and PC cell lines. In PC cell lines, MTT assays, colony formation assays, and flow cytometry were used to investigate the effects of TUG1 on proliferation, cell cycle regulation, and apoptosis. Moreover, we established a xenograft model to assess the effect of TUG1 on tumor growth in vivo. The molecular mechanism of potential target genes was detected through nuclear separation experiments, RNA immunoprecipitation (RIP), chromatin immunoprecipitation assays (ChIP), and other experimental methods. Results The findings suggest that the abnormally high expression of TUG1 in PC tissues was associated with tumor size and pathological stage. Knockdown of TUG1 blocked the cell cycle and accelerated apoptosis, thereby inhibiting the proliferation of PC cells. In addition, RIP experiments showed that TUG1 can recruit enhancer of zeste homolog 2 (EZH2) to the promoter regions of Rho family GTPase 3 (RND3) and metallothionein 2A (MT2A) and inhibit their expression at the transcriptional level. Furthermore, ChIP experiments demonstrated that EZH2 could bind to the promoter regions of RND3 and MT2A. The knockdown of TUG1 reduced this binding capacity. Conclusion In conclusion, our data suggest that TUG1 may regulate the expression of PC-associated tumor suppressor genes at the transcriptional level and these may become potential targets for the diagnosis and treatment of PC.

Bingqing Hui,1,2,* Yetao Xu,3,* Benpeng Zhao,4,* Hao Ji,1,2 Zhonghua Ma,1,2 Shufen Xu,1,2 ZhenYu He,1,5 Keming Wang,1,2 Jianwei Lu6 1Department of Oncology, Second Affiliated Hospital, Nanjing Medical University, Nanjing 210000, Jiangsu, China; 2Department of Oncology, Second Clinical Medical College of Nanjing Medical University, Nanjing 210000, Jiangsu, China; 3Department of Obstetrics and Gynecology, The First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, Jiangsu, China; 4Basic Medicine Faculty of Shanghai Jiaotong University, Core Facility of Basic Medical Sciences, Shanghai 200000, China; 5Department of General Surgery, Second Affiliated Hospital, Nanjing Medical University, Nanjing 210000, Jiangsu, China; 6Department of Medical Oncology, Jiangsu Cancer Hospital, Jiangsu Institute of Cancer Research, Nanjing Medical University Affiliated Cancer Hospital, Nanjing 210000, Jiangsu, China *These authors contributed equally to this work Background: Long noncoding RNAs (lncRNAs) are involved in various human diseases, including cancers. However, their mechanisms remain undocumented. We investigated alterations in lncRNA that may be related to pancreatic cancer (PC) through analysis of microarray data. Methods: In the present study, quantitative real-time PCR analysis was used to examine the expression of taurine upregulated 1 (TUG1) in PC tissue samples and PC cell lines. In PC cell lines, MTT assays, colony formation assays, and flow cytometry were used to investigate the effects of TUG1 on proliferation, cell cycle regulation, and apoptosis. Moreover, we established a xenograft model to assess the effect of TUG1 on tumor growth in vivo. The molecular mechanism of potential target genes was detected through nuclear separation experiments, RNA immunoprecipitation (RIP), chromatin immunoprecipitation assays (ChIP), and other experimental methods. Results: The findings suggest that the abnormally high expression of TUG1 in PC tissues was associated with tumor size and pathological stage. Knockdown of TUG1 blocked the cell cycle and accelerated apoptosis, thereby inhibiting the proliferation of PC cells. In addition, RIP experiments showed that TUG1 can recruit enhancer of zeste homolog 2 (EZH2) to the promoter regions of Rho family GTPase 3 (RND3) and metallothionein 2A (MT2A) and inhibit their expression at the transcriptional level. Furthermore, ChIP experiments demonstrated that EZH2 could bind to the promoter regions of RND3 and MT2A. The knockdown of TUG1 reduced this binding capacity. Conclusion: In conclusion, our data suggest that TUG1 may regulate the expression of PC-associated tumor suppressor genes at the transcriptional level and these may become potential targets for the diagnosis and treatment of PC. Keywords: LncRNA, ncRNA, regulate, mechanism, cancer, EZH2, transcriptional level, tumor suppressor genes

Hui B, Xu Y, Zhao B, et al. Onco Targets Ther. 2019;12:1043–1057. The Editor and Publisher of OncoTargets and Therapy wish to retract the published article. Concerns were raised over alleged image duplication between Figures 2F and 6E. Specifically: Figure 2F, panel AsPC-1 Empty vector appears to have been duplicated with Figure 6E, panel BxPC-3 si-NC. The author responded to our queries but were unable to provide a satisfactory explanation for the alleged duplication and could not provide the original data for the study. The Editor requested for the article to be retracted and the authors agreed with this decision. The authors wish to apologise for this error. Our decision-making was informed by our policy on publishing ethics and integrity and the COPE guidelines on retraction. The retracted article will remain online to maintain the scholarly record, but it will be digitally watermarked on each page as “Retracted”. This retraction relates to this paper

Cholangiocarcinoma (CCA) is the second most malignant tumor of the hepatobiliary system. Due to its cumbersome early diagnosis and rapid progression, chemotherapy has become the main treatment option. Primary drug resistance is a major cause of the poor efficacy of chemotherapeutic drugs. Therefore, it is considered urgent to explore new drugs to overcome primary drug resistance of CCA. Western blot and qRT-PCR assays were used to assess the expression of myotrophin (MTPN) and microRNA-885-5p (miR-885-5p) in CCA tissues and cells. The viability of CCA cells treated with arsenic trioxide (ATO), 5-fluorouracil (5-Fu) and cisplatin (CDDP) was analyzed using a CCK-8 assay. A luciferase reporter assay was used to assess the interaction between miR-885-5p and MTPN. Kaplan-Meier analyses were used for survival assessments. We found that ATO can reduce the resistance of CCA cells to 5-Fu and CDDP and promote the killing effect of 5-Fu and CDDP. Low-dose ATO showed an anti-drug-resistance effect through up-regulation of the expression of miR-885-5p. Combined with sequencing results and database predictions, we found that MTPN may serve as a direct target of miR-885-5p. After MTPN knockdown, the sensitivity of CCA cells to 5-FU and CDDP was increased. Finally, we found that ATO can reverse chemotherapy resistance induced by overexpression of MTPN. Our data indicate that the ATO/miR-885-5p/MTPN axis may serve as a target for improving the sensitivity of CCA cells to chemotherapy.

Background LncRNA can regulate gene at various levels such as apparent genetics, alternative splicing, and regulation of mRNA degradation. However, the molecular mechanism of LncRNA in cholangiocarcinoma is still unclear. This deserves further exploration. Methods We investigated the expression of AGAP2-AS1 in 32 CCA tissues and two CCA cell lines. We found a LncRNA AGAP2-AS1 which induced by SP1 has not been reported in CCA, and Knockdown and overexpression were used to investigate the biological role of AGAP2-AS1 in vitro. CHIP and RIP were performed to verify the putative targets of AGAP2-AS1. Results AGAP2-AS1 was significantly upregulated in CCA tumor tissues. SP1 induced AGAP2-AS1 plays an important role in tumorigenesis. AGAP2-AS1 knockdown significantly inhibited proliferation and caused apoptosis in CCA cells. In addition, we demonstrated that AGAP2-AS1 promotes the proliferation of CCA. Conclusions We conclude that the long non-coding RNA AGAP2-AS1 plays a role in promoting the proliferation of cholangiocarcinoma.

Additional file 1: Table S1. The list of primers and the sequence of siRNAs.

Background: LncRNA can regulate gene at various levels such as apparent genetics, alternative splicing, and regulation of mRNA degradation. However, the molecular mechanism of LncRNA in cholangiocarcinoma is still unclear. This deserves further exploration. Methods:We investigated the expression of AGAP2-AS1 in 32 CCA tissues and two CCA cell lines. We found a LncRNA AGAP2-AS1 which induced by SP1 has not been reported in CCA, and Knockdown and overexpression were used to investigate the biological role of AGAP2-AS1 in vitro. CHIP and RIP were performed to verify the putative targets of AGAP2-AS1. Results:AGAP2-AS1 was significantly upregulated in CCA tumor tissues.SP1 induced AGAP2-AS1 plays an important role in tumorigenesis. AGAP2-AS1 knockdown significantly inhibited proliferation and caused apoptosis in CCA cells. In addition, we demonstrated that AGAP2-AS1 promotes the proliferation of CCA. Conclusions: We conclude that the long non-coding RNA AGAP2-AS1 plays a role in promoting the proliferation of cholangiocarcinoma.

Backgroud: In recent years, the incidence of cholangiocarcinoma (CCA) has increased, and it has become the second most malignant tumor of the hepatobiliary system. Chemotherapy has become the main treatment for cholangiocarcinoma due to its difficulty in diagnosis and rapid progress. Primary drug resistance is the main reason for the poor efficacy of chemotherapeutic drugs. Methods Western blot and quantitative real-time PCR assays were used to detect the expression levels of myotrophin (MTPN) and microRNA-885-5p(miR-885-5p) in CCA tissues and cells; Cell viabilities treated with arsenic trioxide (ATO), 5-fluorouracil (5-Fu) and cisplatin (CDDP) were analyzed by a CCK-8 kit. Luciferase reporter assay detected the relationship between miR-885-5p and MTPN. Kaplan-Meier analysis showed survival time curve. Results We found that ATO can reduce the resistance of CCA cells to 5-Fu and CDDP and promote the killing effect of 5-Fu and CDDP. Low-dose ATO played an anti-drug-resistance role through up-regulating the expression of miR-885-5p. Combined with sequencing results and database prediction, we found that MTPN was a direct target gene of miR-885-5p. The sensitivity of CCA cells to 5-Fu and CDDP was increased after MTPN was knocked out. After MTPN knockout, the sensitivity of cholangiocarcinoma cells to 5-FU and CDDP was increased. ATO can reverse chemotherapy resistance induced by overexpression of MTPN. Conclusions Our study suggests that the ATO/miR-885-5p/MTPN axis is a potential therapeutic strategy for improving the sensitivity of CCA to chemotherapy.

Introduction LncRNA MIR503HG has been reported to participate in liver cancer and ALK-negative anaplastic large-cell lymphoma, while its role in non-small cell lung cancer (NSCLC) is unknown. We therefore investigated the functions of lncRNA MIR503HG in NSCLC. Methods MIR503HG expression in paired cancer and non-cancer tissues from NSCLC patients was analyzed by RT-qPCR. The interaction between cyclin D1 and MIR503HG was analyzed by overexpression experiments. Cell cycle analysis was performed by flow cytometry. Cell proliferation was analyzed by CCK-8 assay. Results MIR503HG was downregulated in NSCLC and low levels of MIR503HG were associated with poor survival. In contrast, cyclin D1 was upregulated in NSCLC, and cyclin D1 and MIR503HG were inversely correlated. In NSCLC cells, overexpression experiments revealed that MIR503HG functioned as an upstream inhibitor of cyclin D1. MIR503HG overexpression led to G1 cell cycle arrest, while overexpression of cyclin D1 attenuated the effects of MIR503HG overexpression. Similarly, MIR503HG overexpression resulted in reduced cell proliferation rate, while overexpression of cyclin D1 caused the increased cell proliferation rate and attenuated effects of MIR503HG overexpression. Conclusion MIR503HG inhibits NSCLC cell proliferation by inducing cell cycle arrest through the downregulation of cyclin D1.

Background: Long non-coding RNA with a length of more than 200 nucleotides, called long non-coding RNA (LncRNAs), lncRNAs can regulate its activity or change its location by interfering with the activity of mRNA and directly binding to proteins. It can also affect the expression of downstream genes by inhibiting RNA polymerase, And as a competitive endogenous RNAs to regulate gene expression, thus playing an important role in the biological process. Colorectal cancer is a malignant lesion of colorectal mucosa caused by environmental, genetic and epigenetic factors. The occurrence of colorectal cancer is a multi-stage, multi-gene process, but its pathogenesis is not fully understood. With the discovery of new molecular and epigenetic mechanisms in CRC, lncRNA has become a biological target for diagnostic, prognostic and therapeutic applications. Methods: Therefore, by screening the GEO database, we found that the expression of Linc01003 in cancer tissues was significantly different from that in paracancerous tissues, and then the expression was detected in 60 pairs of colorectal cancer tissues, five colorectal cancer cell lines and normal intestinal epithelial cell lines. In order to verify the phenotype of linc01003 by knockdown and overexpression experiments, and to explore the mechanism of linc01003 regulating mechanism of proliferation and apoptosis, the upstream transcription factors were predicted by database and verified by related experiments. The related downstream targets were screened by high throughput sequencing and verified by qRT-PCR, Western blot, RIP, ChIP, RNA-FISH and other experiments. Results: Highly expressed linc01003 is closely related to more advanced pathological stages and shorter life cycles.The results showed that the expression of LncRNA Linc01003 induced by transcription factor FOXP1 was significantly increased in colorectal cancer tissues and cells. Knockdown of Linc01003 in colorectal cells could significantly inhibit cell proliferation and induce apoptosis, while overexpression could promote cell proliferation. This can also be proved in animal experiments. After high throughput sequencing, it was found that the expression of crude oncogene HKDC1 was up-regulated and the expression of tumor suppressor gene DIC was down-regulated. Then through bioinformatics prediction and RIP experiments, it was proved that linc01003 binds to EZH2 and AGO2 proteins. CHIP experiments showed that Linc01003 could bind EZH2 to participate in the transcriptional regulation of HKDC1. Luciferase reporter gene confirmed that linc01003 competitive adsorption of miR-106b-5p promoted SLC25A10( DIC) expression. Therefore, we conclude that long non-coding RNA Linc01003 with high expression is associated with later stage and worse prognosis, and transcription factor FOXP1 can induce high expression of long non-coding RNA Linc01003. The high expression of linc01003 promotes the proliferation and inhibits the apoptosis of colorectal cancer cells. Lin01003 promotes the proliferation of colorectal cancer cells through EZH2/HKDC1 and miR-106b-5p/DIC regulatory axes. Conclusions: Therefore, we conclude that the transcription factor FOXP1 induced linc01003 promotes proliferation and inhibits apoptosis of colorectal cancer cells through EZH2/HKDC1 and miR-106b-5p/DIC regulatory axis . It is expected to be a diagnostic marker for CRC and a potential biotherapeutic target. Funding Statement: This study was supported by grants from the National Natural Sciences Foundation of China (grant no. 81772603), Key talents of young medicine in Jiangsu Province(grant no.QNRC2016662), Nanjing Health Science and Technology Development Medical Key Technology Development Project (grant no.YKK18190). Declaration of Interests: The authors declare no competing financial interests. Ethics Approval Statement: All patients provided written informed consent, and all the experiments were approved by the Research Ethics Committee of Nanjing Medical University. This study was conducted in strict accordance with the guidelines of the National Institutes of Health (NIH) on the use of laboratory animals. Our plan has been approved by the Animal Experimental Ethics Committee of Nanjing Medical University.

Misregulation of vascular endothelial growth factor A (VEGF‑A) has been implicated in numerous types of ovarian disease, such as polycystic ovarian syndrome, ovarian hyperstimulation syndrome, endometriosis and ovarian cancer. VEGF regulates blood vessel permeability and angiogenesis. In our previous study, VEGF‑regulated gene expression was profiled in the uterus of a transgenic mouse model with repressed VEGF expression, which indicated that VEGF is an important regulator in controlling gene expression in the uterus. The anti‑Müllerian hormone (AMH) is expressed by ovarian granulosa cells (GCs) and acts through its type 2 receptor, AMH receptor 2 (AMHR2). Serum AMH levels are used to predict ovarian reserves and the small antral follicles contribute markedly to the serum AMH level. AMH recruits primordial follicles and inhibits excessive follicular development by follicular stimulating hormone (FSH). However, AMH may be influenced by suppression of gonadotrophin secretion and VEGF inhibition. In the current study, human primary ovarian GCs were isolated from ovarian follicle fluid of in vitro fertilization/intracytoplasmic sperm injection cycles (IVF/ICSI). It was identified that the FSH receptor was consistently expressed in the isolated cells. VEGF‑A treatment stimulated AMHR2 overexpression at the gene and protein levels. In addition, VEGF induced AMHR2 expression on the surface of the isolated GCs from mature follicles. The VEGF treatment was also performed in an ovarian granulosa‑like cell line, KGN. AMH and AMHR2 are co‑expressed in normal GCs; however, as a result of VEGF misregulation, AMHR2 overexpression increases AMH binding, which may attenuate follicular or oocyte maturation. However, the associated function and underlying mechanism requires further investigation.

Aim To study the value of ultrasound-guided core needle biopsy (CNB) in the diagnosis of T3 or T4 stage laryngeal and hypopharyngeal cancer, which is difficult by routine methods. Patients and methods Nineteen cases of T3 or T4 stage laryngeal or hypopharyngeal carcinoma with abnormal pharyngeal sensitivity, severe dyspnea, submucous cancer recurrence, cardiovascular and pulmonary dysfunction were reviewed retrospectively from October 2012 to October 2014 in the Yuhuangding Hospital of Qingdao University. Ultrasound-guided coarse needle biopsies were used on primary lesions after assessing the patients with neck-enhanced computed tomography (CT) or magnetic resonance imaging (MRI) scan(s). The clinical value of ultrasound-guided CNB in the diagnosis of laryngeal and hypopharyngeal cancer was analyzed. Results All patients underwent successful pathological diagnosis by ultrasound-guided CNB without any serious complications. Dyspnea, cardiovascular and pulmonary dysfunction did not deteriorate. Conclusion Ultrasound-guided CNB is a highly safe and efficient method for the pathological diagnosis of T3 or T4 stage laryngeal and hypopharyngeal cancer. It should be used especially when the fiberoptic or laryngoscope biopsy are of high risk.

The efficacy of immunization with DNA plasmids for single truncated carcinoembryonic antigen (CEA) peptide or triple repeated CEA peptides in mice was evaluated. A DNA fragment the truncated CEA gene (nucleotide 625-667) encoding two helper T lymphocyte (HTL) epitopes was amplified by PCR and cloned for generating recombinant plasmids for single CEA(625-667) (pcDNA-CEA(625-667)) or triple CEA(625-667) (pcDNA-triCEA(625-667)), respectively. Subsequently, groups of BALB/c female mice were intramuscularly injected with pcDNA-CEA(625-667,) pcDNA-triCEA(625-667) or control pcDNA3.0 vector, respectively. Ten days after the last immunization, the frequency of splenic CD4(+) and CD8(+) T cells in the mice was determined by flow cytometry. The antigen-specific proliferation of splenic T cells and cytokine production ex vivo were analyzed by (3)H-TdR uptake and cytokine ELISA, respectively. The levels of serum antibodies against CEA in the mice were detected by Western blot and ELISA. Although immunization with plasmid for the CEA(625-667) peptide(s) did not alter the frequency of CD4(+) and CD8(+) T cells in mice, vaccination with plasmid for CEA peptide induced strong antigen-specific T cell proliferation, particularly for the plasmid encoding the triple-repeated CEA peptides, accompanied by significantly elevated levels of IFN-γ secreted by T cells from the mice immunized with triple-repeated peptides. Furthermore, immunization with the plasmid for CEA peptide stimulated higher levels of antigen-specific antibody responses in mice. Vaccination with the plasmid for the triple repeated CEA peptides induced stronger Th1 responses. Our findings may be useful for the development of effective DNA vaccine for the immunotherapy of cancer.

A two-step synthesis of N-heterocyclic carbene (NHC) precatalysts by diastereoselective or enantioselective lithiation of pyrrolo[1,2-c]imidazol-3-ones followed by POCl(3)-induced salt formation is described. The resulting 3-chloro-pyrroloimidazol(in)ium salts may be coordinated to palladium(II) upon NHC generation with t-BuLi at low temperature. The method may facilitate exploitation of these compounds as chiral organocatalysts or ligands in metal catalysis.

Several 2- and 2,9-substituted 1,10-phenanthrolines are reduced asymmetrically for the first time using a Hantzsch dihydropyridine in the presence of BINOL-derived phosphoric acid catalysts. The best results are obtained with phenanthrolines bearing unbranched or nitrogen-containing alkyl groups in the 2- or 2,9-positions, which afford chiral octahydrophenanthrolines in a range of yields (40-88%) and good to excellent levels of enantiomeric purity (78-99% ee).

Variously substituted 1,10-phenanthrolines are reduced to octahydrophenanthrolines in moderate to good yields with NaBH3CN in acetic acid/methanol. The exact solvent composition is important to avoid the formation of tetrahydrophenanthrolines and N-alkylated byproducts, and to optimize the formation of octahydrophenanthrolines. Resolution of a racemic reduction product gives an enantiomerically pure C2-symmetric diamine from which the corresponding rigid benzimidazolylidene is prepared, whereas reduction of chiral phenanthrolines derived from bicyclic ketones affords diastereomerically pure diamines, which may also be converted to benzimidazolylidenes.

A two-step synthesis of N-heterocyclic carbene (NHC) precatalysts by diastereoselective or enantioselective lithiation of pyrrolo[1,2-c]imidazol-3-ones followed by POCl3-induced salt formation is described. The resulting 3-chloro-pyrroloimidazol(in)ium salts may be coordinated to palladium(II) upon NHC generation with t-BuLi at low temperature. The method may facilitate exploitation of these compounds as chiral organocatalysts or ligands in metal catalysis.