BACKGROUND:Although microRNAs (miRNAs) play essential roles in spermatogenesis, little is known about seminal plasma miRNAs in infertile men. We investigated the profile of seminal plasma miRNAs in infertile men to identify miRNAs that are altered in infertility; we then evaluated their diagnostic value.

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Methyltransferase-like 1 (METTL1) mediated 7-methylguanosine (m7G) is crucial for the regulation of chemoresistance in cancer treatment. However, the role of METTL1 in regulating chemoresistance of colon cancer (CC) cells to cisplatin is still unclear. This study established the cisplatin-resistant CC (CR-CC) cells and found that METTL1 was low-expressed in CR-CC cells compared to their paired cisplatin-sensitive CC (CS-CC) cells. Besides, overexpressed METTL1 enhanced the cytotoxic effects of cisplatin on CR-CC cells. In addition, miR-149-3p was the downstream target of METTL1, which could be positively regulated by METTL1. Further results validated that miR-149-3p was low-expressed in CR-CC cells comparing to the CS-CC cells. In addition, the promoting effects of overexpressed METTL1 on cisplatin induced CR-CC cell death were abrogated by synergistically knocking down miR-149-3p. Furthermore, S100A4/p53 axis was the downstream target of METTL1 and miR-149-3p, and either overexpressed METTL1 or miR-149-3p increased p53 protein levels in CR-CC cells, which were reversed by upregulating S100A4. Similarly, the promoting effects of overexpressed METTL1 on cisplatin-induced CR-CC cell death were abrogated by overexpressing S100A4. Taken together, overexpression of METTL1 sensitized CR-CC cells to cisplatin by modulating miR-149-3p/S100A4/p53 axis.

The nucleotide sequence of hrpNEcc DNA, cloned from Erwinia carotovora subsp. carotovora strain Ecc71, reveals a coding region of 1,068 bp which matches the size of hrpNEcc transcripts. hrpNEcc is predicted to encode a glycine-rich protein of approximately 36 kDa. Like the elicitors of the hypersensitive reaction (HR) produced by E. chrysanthemi (HarpinEch) and E. amylovora (HarpinEa), the deduced 36-kDa protein does not possess a typical signal sequence, but it contains a putative membrane-spanning domain. In Escherichia coli strains overexpressing hrpNEcc, the 36-kDa protein has been identified as the hrpNEcc product by Western blot analysis using anti-HarpinEch antibodies. The 36-kDa protein fractionated from E. coli elicits the HR in tobacco leaves. Moreover, a HrpN- and RsmA- double mutant (RsmA = regulator of secondary metabolites) does not produce this 36-kDa protein or elicit the HR, although this strain, like the RsmA- and HrpN+ bacteria, overproduces extracellular enzymes and macerates celery petioles. These observations demonstrate that hrpNEcc encodes the elicitor of the HR, designated HarpinEcc. The levels of hrpNEcc transcripts are affected in both RsmA+ and RsmA- strains by media composition and carbon sources, although the mRNA levels are substantially higher in the RsmA- strains. The expression of hrpNEcc in Ecc71 is cell density dependent and is activated by the quorum-sensing signal, N-(3-oxohexanoyl)-L-homoserine lactone (OHL). By contrast, hrpNEcc expression in an RsmA- strain is independent of cell density, and substantial expression occurs in the absence of OHL. The effects of cultural conditions and the occurrence of putative cis-acting sequences, such as consensus sigma 54 promoters and an hrp promoter upstream of the transcriptional start site, indicate that the production of HarpinEcc in wild-type RsmA+ E. carotovora subsp. carotovora is tightly regulated. These observations, taken along with the finding that the HR is caused by RsmA- mutants but not by RsmA+ strains (Cui et al., 1996, Mol. Plant-Microbe Interact. 9:565-573), strongly support the idea that the inability of the wild-type pectolytic E. carotovora subsp. carotovora to elicit the HR is due to the lack of a significant level of HarpinEcc production.

Tumor-induced expansion of Tregs is a significant obstacle to cancer immunotherapy. However, traditional approaches to deplete Tregs are often inefficient, provoking autoimmunity. We show here that administration of IL-27-expressing recombinant adeno-associated virus (AAV-IL-27) significantly inhibits tumor growth and enhances T cell responses in tumors. Strikingly, we found that AAV-IL-27 treatment causes rapid depletion of Tregs in peripheral blood, lymphoid organs, and - most pronouncedly - tumor microenvironment. AAV-IL-27-mediated Treg depletion is dependent on IL-27 receptor and Stat1 in Tregs and is a combined result of CD25 downregulation in Tregs and inhibition of IL-2 production by T cells. In combination with a GM-CSF vaccine, AAV-IL-27 treatment not only induced nearly complete tumor rejection, but also resulted in amplified neoantigen-specific T cell responses. AAV-IL-27 also dramatically increased the efficacy of anti-PD-1 therapy, presumably due to induction of PD-L1 in T cells and depletion of Tregs. Importantly, AAV-IL-27 therapy did not induce significant adverse events, partially due to its induction of IL-10. In a plasmacytoma mouse model, we found that IL-10 was required for AAV-IL-27-mediated tumor rejection. Thus, our study demonstrates the potential of AAV-IL-27 as an independent cancer therapeutic and as an efficient adjuvant for cancer immunotherapy.

A major paradigm in cancer immunotherapy is to use checkpoint inhibitors to break regulatory mechanisms that usually guard the host against autoimmune diseases. CTLA-4-targeting immunotherapy was the first example that help to establish this paradigm. However, the clinically tested anti-CTLA-4 antibodies exhibit suboptimal efficacy but high toxicity. Recent studies have demonstrated that immunotherapy-related adverse events (irAE) and the cancer immunotherapeutic effect (CITE) represent distinct and therapeutically separable activities of anti-CTLA-4 antibodies. The former is attributable to inactivation of CTLA-4 checkpoint, while the latter is due to selective depletion of regulatory T cells (Treg) in tumor microenvironment. Here we argue that for safer and more effective CTLA-4-targeting immune therapy, one should preserve rather than inhibit the CTLA-4 checkpoint while enhancing the efficacy and selectivity of Tregdepletion in tumor microenvironment.

Abstract2D black phosphorus (BP) nanosheets and BP quantum dots (BPQD), as two main material styles of BP, are widely used in the biomedical filed. However, few stimuli‐responsive BP nanocarriers are reported to meet the need of nanomedicine. Herein, near‐infrared region/reactive oxygen species (NIR/ROS) sensitive BPQD vesicles (BPNVs) are prepared by self‐assembly of amphiphilic BPQDs grafted with polyethylene glycol and ROS sensitive poly(propylene sulfide) (PPS). BPNVs exhibit enhanced photo‐absorption in the NIR region, and have high loading efficiency of immunoadjuvant CpG oligodeoxynucleotides (CpG ODNs) in the cavity of the BPNVs. Upon NIR laser irradiation, high levels of ROS are generated from BPNVs to trigger the change of hydrophobic PPS to hydrophilic polymers, leading to disassembly of the vesicles to BPQDs. In this manner, the BPNVs show synergistic photodynamic therapy combined with immunotherapy, due to simultaneous release of small BPQDs with deep tumor penetration and CpG with enhanced immunotherapy. The BPNVs‐CpG achieves potent photodynamic immunotherapy in vivo, in addition to block distant tumor growth and metastasis.

SummaryHepatocellular carcinoma (HCC) is one of the most common cancers worldwide, and is often diagnosed at an advanced stage. We have investigated α-fetoprotein (AFP) as a tumor-associated antigen for HCC. We identified major histocompatibility complex class I-restricted peptide epitopes derived from AFP and studied CD8 + T-cell responses in vivo and in vitro in ongoing immunotherapy studies. Helper T cells are of critical importance in shaping the immune response; therefore, we investigated the frequency and function of AFP-specific CD4 + T cells in the general population and among HCC patients. CD4 + T-cell responses were assessed by direct ex vivo multicytokine enzyme-linked immunospot assay and by measurement of cytokine levels using a multicytokine assay. Our analysis indicates that healthy donors have very low frequencies of AFPspecific CD4 + T-cell responses, which are of T H 1 type, detectable ex vivo. In contrast, these T cells were either reduced or eliminated in HCC patients at advanced stages of disease. To better activate these cells, we compared the stimulatory capacity of both AFP protein-fed and AdVhAFP-engineered dendritic cells (DC). Healthy donors have CD4 + T-cell responses, which were activated in response to AFP protein-fed DC whereas HCC patients do not demonstrate significant responses to AFP protein. AdVhAFP-transduced DC were capable of activating higher frequency T H 1 CD4 + responses to AFP in both healthy donors and AFP-positive HCC patients. Importantly, CD4 + T-cell cytokine expression profiles were skewed towards interleukin-2 and interferon-γ production when activated by adenovirally engineered DC, which has therapeutic implications for vaccination efforts. Financial Disclosure: One of the authors is a coinventor on a recently granted US Patent (L. H. B., US Patent No. 7,098,306, "Method and compositions for treating hepatocellular cancer," granted August 2006), which covers some of the MHC class I peptides used in a recently published clinical trial testing AFP class I peptides pulsed onto autologous DC. The intellectual property is owned by the University of California, has not been licensed, and there has been no financial interest for the inventors, to date. The HCC patient PBMC samples used in this work were from that clinical trial. The remaining authors have declared there are no financial conflicts of interest related to this work. [1][2][3] However, the majority of cases are detected at advanced stages, which, even if adequately treated locally, relapse systemically, and HCC recurs in 75% to 100% of patients at 5 years. 4 There are no effective systemic therapies for this disease. 5,6 This leads to a 9% 5-year survival rate after diagnosis in the United States, the second lowest survival rate for any type of cancer. 7 NIH Public Accessα-fetoprotein (AFP) is the most common serum protein during embryonic development, and is a physiologic counterpart of adult serum albumin. Suppression of AFP synthesis occurs shortly after birth. However, AFP mRNA can be detected in hum...