[Circular RNA ame_circ_000115 regulates expression of genes in larval gusts of Apis mellifera ligustica stressed by Ascosphaera apis].

Circular RNAs (circRNAs) are a new class of non-coding RNAs, which have been confirmed to regulate insect gene expression and immune response through multiple manners such as competing endogenous RNA (ceRNA) regulatory network. Currently, function of circRNA in honey bee immune response remains unclear. In this study, PCR and Sanger sequencing were performed to validate the back splicing (BS) site of ame_circ_000115 (in short ac115). RT-qPCR was used to detect the expression profile of ac115 in larval guts of Apis mellifera ligustica stressed by Ascosphaera apis. Dual-luciferase reporter gene assay was conducted to verify the binding relationship between ac115 and ame-miR-13b. Interference of ac115 in larval guts was carried out by feeding specific siRNA, followed by determination of the effect of ac115 interference on expression of six genes relevant to host immune response. The results confirmed the existence of BS site within ac115. Compared with the un-inoculated group, the expression of ac115 in 4-day-old larval gut of the A. apis-inoculated group was up-regulated with extreme significance (P < 0.000 1), while that in 5- and 6-day-old larval guts were significantly up-regulated (P < 0.05). The brightness of specific band for ac115 in 4-, 5- and 6-day-old larval guts of the siRNA-circ_000115-fed group gradually became weak, whereas that of the siRNA-scrambl-fed group was pretty high without obvious variation. Compared with that of the siRNA-scramble-fed group, the expression of ac115 in 4-day-old larval gut of the siRNA-circ_000115-fed group was significantly down-regulated (P < 0.05), whereas that of the 5- and 6-day-old larval guts were down-regulated with extreme significance (P < 0.001). Ame-miR-13b was truly existed and expressed in A. m. ligustica larval guts, and there was true binding relationship between ac115 and ame-miR-13b. Compared with that of the siRNA-scramble-fed group, the expression of antimicrobial peptide genes hymenoptaecin and abaecin in 6-day-old larval gut of the siRNA-circ_000115-fed group was significantly up-regulated (P < 0.05), while that of ecdysone receptor (Ecr) was down-regulated with extreme significance (P < 0.01). These results indicate that ac115 is truly expressed in A. m. ligustica larval guts, BS site truly exists within ac115, and effective interference of ac115 in A. m. ligustica larval guts can be achieved via feeding siRNA. Moreover, ac115 potentially regulates Ecr expression through adsorption of ame-miR-13b and expression of hymenoptaecin and abaecin using a non-ceRNA manner, further participating in host stress-response.

CircRNA-regulated immune responses of asian honey bee workers to microsporidian infection.

Nosema ceranae is a widespread fungal parasite for honey bees, causing bee nosemosis. Based on deep sequencing and bioinformatics, identification of circular RNAs (circRNAs) in Apis cerana workers' midguts and circRNA-regulated immune response of host to N. ceranae invasion were conducted in this current work, followed by molecular verification of back-splicing sites and expression trends of circRNAs. Here, 10185 and 7405 circRNAs were identified in the midguts of workers at 7 days (AcT1) and 10 days (AcT2) post inoculation days post-inoculation with N. ceranae. PCR amplification result verified the back-splicing sites within three specific circRNAs (novel_circ_005123, novel_circ_007177, and novel_circ_015140) expressed in N. ceranae-inoculated midgut. In combination with transcriptome data from corresponding un-inoculated midguts (AcCK1 and AcCK2), 2266 circRNAs were found to be shared by the aforementioned four groups, whereas the numbers of specific ones were 2618, 1917, 5691, and 3723 respectively. Further, 83 52) differentially expressed circRNAs (DEcircRNAs) were identified in AcCK1 vs. AcT1 (AcCK2 vs. AcT2) comparison group. Source genes of DEcircRNAs in workers' midgut at seven dpi were involved in two cellular immune-related pathways such as endocytosis and ubiquitin mediated proteolysis. Additionally, competing endogenous RNA (ceRNA) network analysis showed that 23 13) DEcircRNAs in AcCK1 vs. AcT1 (AcCK2 vs. AcT2) comparison group could target 18 14) miRNAs and further link to 1111 (1093) mRNAs. These target mRNAs were annotated to six cellular immunity pathways including endocytosis, lysosome, phagosome, ubiquitin mediated proteolysis, metabolism of xenobiotics by cytochrome P450, and insect hormone biosynthesis. Moreover, 284 164) internal ribosome entry site and 54 26) ORFs were identified from DEcircRNAs in AcCK1 vs. AcT1 (AcCK2 vs. AcT2) comparison group; additionally, ORFs in DEcircRNAs in midgut at seven dpi with N. ceranae were associated with several cellular immune pathways including endocytosis and ubiquitin-mediated proteolysis. Ultimately, RT-qPCR results showed that the expression trends of six DEcircRNAs were consistent with those in transcriptome data. These results demonstrated that N. ceranae altered the expression pattern of circRNAs in A. c. cerana workers' midguts, and DEcircRNAs were likely to regulate host cellular and humoral immune response to microsporidian infection. Our findings lay a foundation for clarifying the mechanism underlying host immune response to N. ceranae infection and provide a new insight into interaction between Asian honey bee and microsporidian.

Realizing Compact Lithium Deposition via Elaborative Nucleation and Growth Regulation for Stable Lithium-Metal Batteries.

Metallic lithium (Li) has been regarded as an ideal candidate for anode materials in next-generation high-energy-density batteries. However, a ubiquitous spongy Li deposition results in low reversibility, huge interfacial impedance, and even safety issues, hindering its practical application. Herein, we proposed a bifunctional electrolyte (BiFE) to avoid the spongy Li deposition, in which lithium nitrate (LiNO3) facilitates a uniform granular Li nucleation via forming a kinetically favorable solid electrolyte interphase and silicon dioxide (SiO2) adsorbs anions to stabilize the electric field distribution near the electrode surface. Such a BiFE provides an even Li+ ion flux for the subsequent growth of electrochemical Li deposition, which was verified by ζ potential, Raman spectra, and specific capacitance characterizations, thus realizing a compact and uniform Li deposition via elaborative nucleation and growth regulation. An improved Li Coulombic efficiency of 99.1% can be achieved within BiFE. When used in Cu∥Li half-cells and Li∥Li symmetric cells, the high Li utilization prolonged the cycling life span to above 300 cycles and 1200 h, respectively. The compact Li deposition also resisted the corrosion of polysulfides to enhance the cycling performance of Li∥S full cells.

Simplifying the Electrolyte Systems with the Functional Cosolvent.

The state-of-the-art electrolytes utilized in lithium-ion batteries are based on liquid carbonates combining a number of additives to fulfill the practical requirements including safety and low temperature. The plenty of components result in the quadruple times of probable radical groups involved into the interfacial reactions, rendering it too difficult to control the surface layer. This work tends to simplify the system with the fluorine-substituted ether as the functional cosolvent to expand the functions of basic electrolytes. The incorporation of this solvent enables the electrolyte to self-extinguish, reduces its freezing point to ∼75 °C lower, and assists in the formation of LiF-rich protective interlayers, resulting in the improvement of the rate capability, cryogenic performance, and cyclic stability for the LiNi1/3Co1/3Mn1/3O2 cathode. This novel design could significantly diminish the amount of necessary additives and possess the acceptable cost, which provides a probability to revitalize the development of liquid electrolytes.

[Treatment outcome and prognosis of 112 patients with nasal and nasopharyngeal peripheral T cell lymphomas].

OBJECTIVE: To retrospectively analyze the treatment outcomes and prognostic factors of nasal and nasopharyngeal peripheral T cell lymphomas (PTCL) patients. METHODS: One hundred and twelve patients with pathologically confirmed nasal and nasopharyngeal PTCL were included, among which 39 were CD56(+) NK/T cell lymphomas. The median pre-treatment disease course was 4 months. 84 were males and 28 females median age was 46 years. The tumors mainly involved nasal cavity (88 cases) and/or nasopharynx (50 cases) and adjacent structures, and 83 cases with extra-cavity diseases. 91.1% of the patients had Ann Arbor I(E)/II(E) diseases. The International Prognostic Indices (IPI) were less than 2 scores in 78.8% of the patients. Seventy two patients received combined chemo-radiotherapy, 32 chemotherapy only, 3 radiotherapy only and 5 no any treatment. RESULTS: Median follow-up duration was 42 months. Chemotherapy achieved a complete remission (CR) rate of 34.4% for initial treatment, and of 65.1% after primary treatment. The local tumor controlled rate was 50.5%, and the median time to tumor progression (TTP) was 11 months. There were evidences of systemic relapse in more than 30% of the patients. The extra-cavity tumors usually had a shorter TTP (r(s) = -0.191, P = 0.024). The progress-free survival and overall survival rates were 38.8% and 52.4% at 3 years, and 34.9% and 44.8% at 5 years respectively. Univariate analysis showed that favorable prognostic factors for survival were pre-treatment course > 3 months, earlier clinical stage, non NK/T lymphoma, no skin involvement, lower IPI, CR after initial chemotherapy, radiotherapy, CR after primary treatment and local tumor controlled. Multivariate analysis showed that, pre-treatment course > 3 months (P = 0.011), non NK/T lymphoma (P = 0.007), CR after initial chemotherapy (P = 0.008) and radiotherapy (P = 0.000) were favorable prognostic factors for survival. CONCLUSIONS: Although most nasal and nasopharyngeal peripheral T-cell lymphomas were diagnosed at early stage diseases, some of them were highly aggressive with poor prognosis, particularly CD56(+) NK/T cell lymphomas. Combination chemo/radiotherapy, though remained principal treatments, more effective therapeutic modalities are expected.

[Clinical features and prognosis of nasal type NK/T cell lymphoma].

OBJECTIVE: To investigate the clinical features, treatment modalities and the prognosis of nasal type NK/T cell lymphoma. METHODS: The data of 39 such patients treated from June 2000 to December 2003 were retrospectively reviewed. Twenty three patients were treated by combined chemoradiotherapy, basing on anthracycline-containing CHOP or similar regimens (median 5 cycles). Eleven patients by chemotherapy alone, 2 by radiotherapy alone and 2 aged patients by palliative chemotherapy or radiotherapy. Radiotherapy was given by high energy photon ray combined with electron beam with a median curative dose of 56 Gy in conventional fractionation. Bivariate correlations and univariate prognostic factors were analyzed. RESULTS: Median follow-up time for the 21 patients who were still alive was 22.5 months. The overall remission rate (RR) after initial treatment was 66.7% (21 CR, 3 PR). Chemotherapy alone got a CR rate of only 37.5%. The overall local control rate was 59.4%. Local relapse rate after curative radiotherapy was 25.0%. Radiotherapy was positively correlated with local control (P = 0.000) and time to disease progression (TTP, P = 0.002). Skin and intestine were among the extranodal relapse sites. Fifteen patients had highly aggressive tumors with a median survival time of only 5 months. Univariate analysis showed that significant favorable survival prognostic factors were: radiotherapy (P = 0.001); lower risk International Prognostic Index (IPI, P = 0.001); complete remission after primary treatment (P = 0.000); pre-diagnostic history > 2 months (P = 0.024); and free of skin involvement (P = 0.034). CONCLUSION: Most of nasal type NK/T cell lymphoma are in early stage when diagnosed. Radiotherapy remains to be the mainstay of treatment. Combined chemoradiotherapy needs further improvement for the progressive disease type. Some patients may have highly aggressive tumors with poor prognosis. Optimal prognostic factors and individualized treatment regimens need to be investigated.