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Severe fever with thrombocytopenia syndrome (SFTS) is caused by a phlebovirus of the Bunyaviridae family, which is designated as SFTS virus (SFTSV). To our knowledge, no efficient SFTSV vaccine exists. Here, we report the identification of a standard virus strain for the eight major SFTSV strains circulating in China for use in evaluating the SFTSV vaccine. Rabbits were immunized with the SFTSV strains and the cross-neutralization capacities of SFTSV anti-sera were determined in microculture cytopathic effect (CPE)-inhibition assays. The mean cross-neutralization capacity of the eight SFTSV anti-sera ranged from 62.4 to 142.6%, compared to autologous strains. The HB29 strain demonstrated strong cross-reactivity with heterologous antibodies, and 33 serum samples from SFTS patients efficiently neutralized HB29, suggesting its broad cross-reactivity. In addition, HB29 demonstrated good replication in Vero and MRC-5 cells (8.0 and 6.0 lg 50% cell culture-infectious dose/mL, respectively) and significant CPE, which satisfied the requirements for a standard virus strain. The HB29 isolate was proven identical to the reported HB29 strain by DNA sequencing, and showed high homology in the S segments with other SFTSV strains (94.8–99.7%). Our results suggest that HB29 may be the best candidate standard strain for use in SFTS vaccine development in China.

Phytoestrogen has previously been proposed as an alternative to hormone replacement therapy. Hispolon has been found to have phytoestrogenic properties. However, the possible effects of hispolon on estrogen receptors and other related molecules remain to be determined. This study was performed mainly to confirm the estrogenic function of hispolon as it relates to estrogen receptors, aromatase, and cyclooxygenase 2 (COX-2). Hispolon was shown to increase the serum 17β-estradiol in vivo. Immunohistochemical staining methods showed that hispolon exhibited a biphasic effect on ERα/β and aromatase expression in MCF-7 cells. Hispolon could also significantly inhibit aromatase activity, assessed using the enzyme-linked immunosorbent assay. Western blotting showed that COX-2 and aromatase could be inhibited by hispolon. These results further prove the phytoestrogenic features of hispolon and explore some pharmacological mechanisms that suggest that hispolon could be useful in the treatment of breast cancers or other gynecologic diseases.

Extremely metal-poor galaxies with metallicity below 10% of the solar value in the local universe are the best analogues to investigating the interstellar medium at a quasi-primitive environment in the early universe. In spite of the ongoing formation of stars in these galaxies, the presence of molecular gas (which is known to provide the material reservoir for star formation in galaxies, such as our Milky Way) remains unclear. Here, we report the detection of carbon monoxide (CO), the primary tracer of molecular gas, in a galaxy with 7% solar metallicity, with additional detections in two galaxies at higher metallicities. Such detections offer direct evidence for the existence of molecular gas in these galaxies that contain few metals. Using archived infrared data, it is shown that the molecular gas mass per CO luminosity at extremely low metallicity is approximately one-thousand times the Milky Way value., Comment: 12 pages, 3 figures, 1 table. Supplementary data at http://www.nature.com/article-assets/npg/ncomms/2016/161209/ncomms13789/extref/ncomms13789-s1.pdf

ObjectivesOur previous studies indicated that the triterpenes from the fruits of Chaenomeles speciosa (Sweet) Nakai (TCS) owned effectively therapeutic effects on gastric ulcer patients and animals, but its mechanisms have not been fully understood. The current study was to further investigate its protective effect on indomethacin (IND)-damaged RGM-1 cells and rats, as well as its mechanisms involved.MethodsThe gastroprotection of TCS was evaluated with IND-induced gastric lesions model in RGM-1 cells and rats. In vitro, the proliferation, migration, mitochondrial viability and apoptosis were assessed. In vivo, ulcer index, ulcer inhibition rate, gastric juice acidity, gastric wall mucus (GWM) and histopathology of gastric mucosa were detected. The gastroprotective effects of TCS through the TFF1-mediated EGF/EGFR and apoptotic pathways were measured by qRT-PCR and Western blot assays.Key findingsThe results demonstrated that TCS had gastroprotective function, which was related to the amelioration in promoting IND-damaged RGM-1 cell proliferation and migration, hoisting gastric juice acidity and GWM, improving ulcer index and ulcer inhibition rate, attenuating the haemorrhage, oedema, epithelial cell loss and inflammatory cell infiltration of gastric mucosa, upregulating PCNA, Bcl-2, Bcl-xl mRNA and TFF1, EGF, p-EGFR, p-Src, pro-caspase-3, pro-caspase-9 protein expressions, mitochondrial viability, mitochondrial cytochrome c concentration and p-EGFR/EGFR, p-Src/Src, Bcl-2/Bax, Bcl-xl/Bad ratioes, downregulating Bax, Bad, Apaf-1 mRNA and cleaved-caspase-3, cleaved-caspase-9, cleaved PARP-1 protein expressions and cytosol cytochrome c concentration.ConclusionsOur present study demonstrated that TCS’s gastroprotective effect was closely connected with boosting TFF1 expression, activating TFF1-mediated EGF/EGFR pathway, thus restraining mitochondrial-dependent apoptosis, which provided new insights into interpreting its underlying mechanism and promised to act as a candidate drug to treat gastric mucosal injury.

To investigate the pharmacokinetics of PEGylated recombinant human endostatin (M2ES) in rhesus monkey. M2ES was administered to rhesus monkeys by intravenous bolus injection, and serum M2ES concentration was determined by a self-developed ELISA assay. Pharmacokinetic parameters were calculated using a non-compartmental model of WinNonlin V2.1A software. The standard curve of self-developed ELISA assay was fitted by four-parameter method. The limit of detection (LOD) and LOQ were 0.3050 ng/mL and 0.9140 ng/mL, respectively. Following IV infusions of M2ES at 0.3, 1, and 3 mg/kg in rhesus monkeys, the serum M2ES concentration-time curve was fitted with a non-compartment model. The pharmacokinetic parameters were evaluated as follows: Terminal elimination half-life (T1/2) of M2ES were 3.30 ± 0.70, 29.50 ± 18.80 and 24.60 ± 8.90 h. Systemic clearance (CLsys) of M2ES were 339.60 ± 66.30, 161.40 ± 18.20 and 260.10 ± 43.70 mL/h/kg. AUC(0-∞) values of M2ES were 909.30 ± 199.60, 6251.00 ± 739.60 and 11758.00 ± 2010.10 ng∙h/mL. The dosage was positively correlated with AUC, and the correlation coefficient r2 = 0.9327. Self-developed ELISA assay could meet the requirements of serum M2ES concentration detection. PEGylation modification substantially expands the circulation time of recombinant human endostatin and effectively improves its pharmacokinetic behavior.

To develop fusion protein of a GnRH Fc fragment and the integrin targeting AP25 antitumor peptide for GnRH receptor-expressing cancer therapy. The LMRAP fusion protein was constructed. A transwell invasion assay was performed. The gene mRNA and protein levels of GnRHR-I, α5β1, and αvβ3 in different cancer cell lines were assessed. Cell proliferation was measured using a cell counting kit-8. An antagonist assay was performed on GnRH receptors. Anti-tumor activity was evaluated with a mouse xenograft tumor model. Immunohistochemistry (IHC) was applied to detect CD31 and CD34 expressions. Pharmacokinetic characteristics were determined with an indirect competition ELISA. The developed bifunctional fusion protein LMRAP not only inhibited HUVEC invasion, but also inhibited proliferation of GnRHR-I, α5β1, and αvβ3 high expression cancer cells. The IC50 for LMRAP in the GnRH receptor was 6.235 × 10−4 mol/L. LMRAP significantly inhibited human prostate cancer cell line 22RV1 proliferation in vivo and in vitro. LMRAP significantly inhibited CD31 and CD34 expressions. The elimination half-life of the fusion protein LMRAP was 33 h in rats. The fusion protein made of a GnRH Fc fragment and the integrin targeting AP25 peptide retained the bifunctional biological activity of GnRHR blocking, angiogenesis inhibition, prolonged half-life and good tolerance., Graphical abstract LMRAP fusion protein, constructed by GnRH Fc fragment and the integrin targeting AP25 antitumor peptide, not only retained the bifunctional activity of GnRH receptor blocking and angiogenesis inhibition, but also prolonged the half-life. This study provides clues for developing a new strategy for GnRH receptor-expressing cancer treatment.Image 1

We report the results of our systematic survey for Galactic 6.7 GHz Class II CH$_3$OH maser emission toward a sample of young stellar objects. The survey was conducted with the Shanghai Tianma Radio Telescope (TMRT). The sample consists of 3348 sources selected from the all-sky \emph{Wide-Field Infrared Survey Explorer (WISE)} point source catalog. We have discussed the selection criteria in detail and the detection results of those at high Galactic latitudes (i.e. $|b|>$ 2$^\circ$) in a previous paper (paper I). Here, we present the results from the survey of those at low Galactic latitudes, i.e. $|b, 46 pages, 9 figures, 6 tables

Trametenolic acid B (TAB) was a lanostane-type triterpenoid isolated from the trametes lactinea (Berk.) Pat. We have previously reported that extract from trametes lactinea (Berk.) Pat and TAB could efficiently improve learning and memory ability of the cerebral ischemia injury rats and suppress mitochondrial-mediated apoptosis in hydrogen peroxide damaged SH-SY5Y cells. However, the potential mechanisms have not been fully understood yet. The current study was to further investigate the protective effect of TAB on oxygen glucose deprivation/reoxygenation (OGD/R)-damaged SH-SY5Y cells and cerebral ischemia/reperfusion (I/R) injury rats, as well as its mechanisms involved. Cell experiments demonstrated that TAB (10, 20 and 40 μg/mL) protected OGD/R-induced SH-SY5Y cell injury by promoting cell proliferation and suppressing LDH leakage; Meanwhile, the results in vivo showed that TAB (20, 40 and 80 mg/kg) might significantly ameliorate the neurological deficit score, cerebral edema, neuronal cell loss and apoptosis, suppress cerebral infarction volume of the cerebral I/R injury rats. Further studies in vitro and in vivo indicated TAB could efficiently reduce OGD/R-damaged SH-SY5Y cell and cerebral I/R rat serum ROS, LDH and MDA levels, elevate SOD, GSH-Px and CAT activities, downregulate miR-10a mRNA and Bax, cytochrome C, cleaved-caspase-3 and cleaved-caspase-9 protein expressions, upregulate p-PIK3CA, p-Akt, p-mTOR, Bcl-2, pro-caspase-9 and pro-caspase-3 protein expressions and p-PIK3CA/PIK3CA, p-Akt/Akt, p-mTOR/mTOR ratios (P

JE vaccination is the most effective and economical method of preventing JE. A live attenuated JE vaccine has been widely used in many countries since 1989, playing an important role in controlling JE outbreaks. However, whether the large-scale use of the live attenuated JE vaccine will lead to the dissemination of the vaccine virus in the environment and whether reversion of the neuroattenuation of the virus will occur during the transmission process remain major concerns for some researchers. To evaluate the transmission of a live attenuated JEV vaccine in mosquitoes and hosts, JE SA14-14-2 attenuated vaccine virus was intrathoracically (i.t.) inoculated into Culex tritaeniorhynchus, a native vector. Subsequently, virus harvested from inoculated mosquitoes was inoculated into pigs, a mammalian reservoir. The virus was isolated from the pigs and passaged once again in Culex tritaeniorhynchus. The genome sequences and virulence of the passaged viruses were then investigated. While a few nucleotide substitutions occurred during passaging, there was no change in the encoded amino acids. After intracerebral (i.c.) inoculation of mice with the vaccine, no pathological effects were observed. In addition, virus virulence remained low after inoculation of suckling mouse brains. These results indicate that vaccination of individuals with the live vaccine will not result in transmission of the live SA14-14-2 vaccine virus through mosquito biting and virus amplified in pigs.

During the past two decades, recombinant human interleukin-12 (rhIL-12) has emerged as one of the most potent cytokines in mediating antitumor activity in a variety of preclinical models and clinical studies. Purity is a critical quality attribute (CQA) in the quality control system of rhIL-12. In our study, rhIL-12 bulks from manufacturer B showed a different pattern in non-reduced SDS-PAGE compared with size-exclusion chromatography (SEC)-HPLC. A small fragment was only detected in non-reduced SDS-PAGE but not in SEC-HPLC. The results of UPLC/MS and N-terminal sequencing confirmed that the small fragment was a 261&ndash, 306 amino acid sequence of a p40 subunit of IL-12. The cleavage occurs between Lys260 and Arg261, a basic rich region. With the presence of 0.2% SDS, the small fragment appeared in both native PAGE and in SEC-HPLC, suggesting that it is bound to the remaining part of the IL-12 non-covalently, and is dissociated in a denatured environment. The results of a bioassay showed that the fractured rhIL-12 proteins had deficient biological activity. These findings provide an important reference for the quality control of the production process and the final products of rhIL-12.

The long-acting growth hormone (LAGH) is a promising alternative biopharmaceutical to treat growth hormone (GH) deficiency in children, and it was developed using a variety of technologies by several pharmaceutical companies. Most LAGH preparations, such as Fc fusion protein, are currently undergoing preclinical study and clinical trials. Accurate determination of bioactivity is critical for the efficacy of quality control systems of LAGH. The current in vivo rat weight gain assays used to determine the bioactivity of recombinant human GH (rhGH) in pharmacopoeias are time-consuming, expensive, and imprecise, and there are no recommended bioassays for LAGH bioactivity in pharmacopoeias. Therefore, we developed a cell-based bioassay for bioactivity determination of therapeutic long-acting Fc-fusion recombinant human growth hormone (rhGH-Fc) based on the luciferase reporter gene system, which is involved in the full-length human GH receptor (hGHR) and the SG (SIE and GAS) response element. The established bioassay was comprehensively validated according to the International Council for Harmonization (ICH) Q2 (R1) guidelines and the Chinese Pharmacopoeia, and is highly precise, time-saving, simple, and robust. The validated bioassay could be qualified for bioactivity determination during the research, development, and manufacture of rhGH-Fc, and other LAGH formulations.

Despite significant advances in health care, outbreaks of infections by enteroviruses (EVs) continue to plague the Asia-Pacific region every year. Enterovirus 71 (EV71) causes hand-foot-and-mouth disease (HFMD), for which there are currently no therapeutics. Here, we report two new antibodies, A9 and D6, that potently neutralize EV71. A9 exhibited a 50% neutralizing concentration (neut50) value of 0.1 nM against EV71, which was 10-fold lower than that observed for D6. Investigation into the mechanisms of neutralization revealed that binding of A9 to EV71 blocks receptor binding but also destabilizes and damages the virus capsid structure. In contrast, D6 destabilizes the capsid only slightly but interferes more potently with the attachment of the virus to the host cells. Cryo-electron microscopy (cryo-EM) structures of A9 and D6 bound with EV71 shed light on the locations and nature of the epitopes recognized by the two antibodies. Although some regions of the epitopes recognized by the two antibodies overlap, there are differences that give rise to dissimilarities in potency as well as in the mechanisms of neutralization. Interestingly, the overlapping regions of the epitopes encompass the site that the virus uses to bind SCARB2, explaining the reason for the observed blocking of the virus-receptor interaction by the two antibodies. We also identified structural elements that might play roles in modulating the stability of the EV71 particles, including particle integrity. The molecular features of the A9 and D6 epitopes unveiled in this study open up new avenues for rationally designing antiviral drugs., IMPORTANCE During the course of viral infections, the human body produces neutralizing antibodies which play a defining role in clearing the virus. From this study, we report two new, highly potent neutralizing antibodies, A9 and D6, against enterovirus 71 (EV71), the causative agent of HFMD. Both antibodies prevent the virus from entering the host cell, a step that is important for establishing a successful infection. A9 destabilizes and damages the virus capsid that forms an outer protective covering around the genome of the virus, while also interfering with virus attachment to the host cells. In contrast, D6 only prevents binding of the virus to its receptor(s). The mechanism of neutralization of A9 is unique and has not been observed before for neutralizing antibodies targeting EVs. The two antibodies that we are reporting in this study have potential to be developed into much-needed therapeutic interventions for treatment of HFMD, outbreaks of which are reported every year in the Asia-Pacific region.

According to the No. 13 Mine of Pingdingshan Coal Co. Ltd., the fault dimension of the study area is calculated based on fractal theory. The four impact factors such as water inflow, water pressure, water inrush coefficient, and fault fractal dimension of 21 water boreholes are used as evaluation indices, and a mathematical model for identifying the water-filled source of the coal seam floor is established by coupling an entropy weight method and fuzzy variable set theory. The model is used to identify the water-filled source of 7 boreholes, which provide a reliable reference for the identification of the water-filled source. According to the calculation result of the entropy weight method, the water volume per unit of the borehole and the groundwater pressure have a significant impact on the water source identification, which accounts for 89.93% of the weight value. In the fuzzy variable set model, when the distance parameter p is 1 and the optimization criterion parameter α is 2, the accuracy of the water sample identification of the water source category to be identified is 85.71%, which is at a high recognition level. The more typical the impact factors selected and the more samples, the accuracy of water-filled source identification is much higher.

We performed a survey in the SiO $J=5\rightarrow4$ line toward a sample of 199 Galactic massive star-forming regions at different evolutionary stages with the SMT 10 m and CSO 10.4 m telescopes. The sample consists of 44 infrared dark clouds (IRDCs), 86 protostellar candidates, and 69 young \HII\ regions. We detected SiO $J=5\rightarrow4$ line emission in 102 sources, with a detection rate of 57\%, 37\%, and 65\% for IRDCs, protostellar candidates, and young \HII\ regions, respectively. We find both broad line with Full Widths at Zero Power (FWZP) $>$ 20 \kms and narrow line emissons of SiO in objects at various evolutionary stages, likely associated with high-velocity shocks and low-velocity shocks, respectively. The SiO luminosities do not show apparent differences among various evolutionary stages in our sample. We find no correlation between the SiO abundance and the luminosity-to-mass ratio, indicating that the SiO abundance does not vary significantly in regions at different evolutionary stages of star formation., 25 pages, 9 figures, 5 tables, accepted for publication in ApJ

We detected carbon-chain molecules (CCMs) HC2n+1N (n=1-3) and C3S in Ku band as well as {high-energy excitation lines including C4H N=9-8, J=17/2-15/2, 19/2-17/2, and CH3CCH J=5-4, K=2 in the 3 mm band toward a starless core called the eastern molecular core (EMC) of L1489 IRS. Maps of all the observed lines were also obtained. Comparisons with a number of early starless cores and WCCC source L1527 show that the column densities of C4H and CH3CCH are close to those of L1527, and the CH3CCH column densities of the EMC and L1527 are slightly higher than those of TMC-1. The EMC and L1527 have similar C3S column densities, but they are much lower than those of all the starless cores, with only 6.5% and 10% of the TMC-1 value, respectively. The emissions of the N-bearing species of the EMC and L1527 are at the medium level of the starless cores. These comparisons show that the CCM emissions in the EMC are similar to those of L1527, though L1527 contains a protostar. Although dark and quiescent, the EMC is warmer and at a later evolutionary stage than classical carbon-chain--producing regions in the cold, dark, quiescent early phase. The PACS, SPIRE, and SCUBA maps evidently show that the L1489 IRS seems to be the heating source of the EMC. Although it is located at the margins of the EMC, its bolometric luminosity and bolometric temperature are relatively high. Above all, the EMC is a rather particular carbon-chain-producing region and is quite significant for CCM science., Comment: 16 pages, 7 figures

We performed a systematic 6.7 GHz Class \uppercase\expandafter{\romannumeral2} methanol maser survey using the Shanghai Tianma Radio Telescope toward targets selected from the all-sky \emph{Wide-Field Infrared Survey Explorer (WISE)} point catalog. In this paper, we report the results from the survey of those at high Galactic latitudes, i.e. $|b|>$ 2$^\circ$. Of 1473 selected \emph{WISE} point sources at high latitude, 17 point positions that were actually associated with 12 sources were detected with maser emission, reflecting the rarity (1$-$2\%) of methanol masers in the region away from the Galactic plane. Out of the 12 sources, 3 are detected for the first time. The spectral energy distribution (SED) at infrared bands shows that these new detected masers occur in the massive star forming regions. Compared to previous detections, the methanol maser changes significantly in both spectral profiles and flux densities. The infrared \emph{WISE} images show that almost all of these masers are located in the positions of the bright \emph{WISE} point sources. Compared with the methanol masers at the Galactic plane, these high-latitude methanol masers provide good tracers for investigating the physics and kinematics around massive young stellar objects, because they are believed to be less affected by the surrounding cluster environment., 27pages, 8 figures, 3 tables. Published in ApJ

Using the new equipment of the Shanghai Tian Ma Radio Telescope, we have searched for carbon-chain molecules (CCMs) towards five outflow sources and six Lupus I starless dust cores, including one region known to be characterized by warm carbon-chain chemistry (WCCC), Lupus I-1 (IRAS 15398-3359), and one TMC-1 like cloud, Lupus I-6 (Lupus-1A). Lines of HC3N J=2-1, HC5N J=6-5, HC7N J=14-13, 15-14, 16-15 and C3S J=3-2 were detected in all the targets except in the outflow source L1660 and the starless dust core Lupus I-3/4. The column densities of nitrogen-bearing species range from 10$^{12}$ to 10$^{14}$ cm$^{-2}$ and those of C$_3$S are about 10$^{12}$ cm$^{-2}$. Two outflow sources, I20582+7724 and L1221, could be identified as new carbon-chain--producing regions. Four of the Lupus I dust cores are newly identified as early quiescent and dark carbon-chain--producing regions similar to Lup I-6, which together with the WCCC source, Lup I-1, indicate that carbon-chain-producing regions are popular in Lupus I which can be regard as a Taurus like molecular cloud complex in our Galaxy. The column densities of C3S are larger than those of HC7N in the three outflow sources I20582, L1221 and L1251A. Shocked carbon-chain chemistry (SCCC) is proposed to explain the abnormal high abundances of C3S compared with those of nitrogen-bearing CCMs. Gas-grain chemical models support the idea that shocks can fuel the environment of those sources with enough $S^+$ thus driving the generation of S-bearing CCMs., Comment: 7 figures, 8 tables, accepted by MNRAS

Leukocyte immunoglobulin (Ig)-like receptors (LILRs), also known as CD85 and immunoglobulin-like transcripts (ILTs), play pivotal roles in regulating immune responses. These receptors define an immune checkpoint that immune therapy can target. Through cis or trans interactions with human leukocyte antigen (HLA)-G, the two most abundantly expressed inhibitory LILRs, LILRB1, and LILRB2 (LILRB1/2, also known as CD85j/d and ILT2/4), are involved in immunotolerance in pregnancy and transplantation, autoimmune diseases, and immune evasion by tumors. Although the discrete domains of LILRB1/2 are clear, the assembly mode of the four extracellular Ig-like domains (D1, D2, D3, and D4) remains unknown. Previous data indicate that D1D2 is responsible for binding to HLA class I (HLA-I), but the roles of D3D4 are still unclear. Here, we determined the crystal structure of the four Ig-like domain LILRB2 and four-domain LILRB1 in complex with HLA-G1. The angles between adjacent domains and the staggered assembly of the four domains suggest limited flexibility and limited plasticity of the receptors during ligand binding. The complex structure of four-domain LILRB1 and HLA-G1 supports the model that D1D2 is responsible for HLA-I binding, while D3D4 acts as a scaffold. Accordingly, cis and trans binding models for HLA-I binding to LILRB1/2 are proposed. The geometries of LILRB1/2 in complex with dimeric and monomeric HLA-G1 suggest the accessibility of the dimeric receptor, which in turn, transduces more inhibitory signals. The assembly of LILRB1/2 and its binding to HLA-G1 could aid in the design of immune regulators and benefit immune interference.

The noncoding sequences in the UTR between the Gene Start signal (GS) and the initial codon AUG from 25 measles virus genomes were compared. The conserved sequences 5′-U(NNNNN)5-8-3′ and 5'-(CNNNNNC)-3′ were found in the genome and the UTR of phosphoprotein (P) and Fusion (F) mRNA. These sequences were named the U-Unit and the C-Unit, respectively. These two sequences are different in the P and F mRNAs. To investigate whether these conserved sequences can be used to regulate expression of foreign proteins in additional transcription units (ATU), a series of minigenomes with various 5′ UTRs were generated. From reporter gene assays and quantitation of gene copies by qPCR, we found that the introduction of the C-unit into the 5′UTR of mRNA can down-regulate the expression of foreign proteins in additional transcription units in vivo.

The 2014 Ebola outbreak in West Africa brought great threat to public health worldwide. There was no approved antiviral therapy or vaccine available to control the disease at that time. Several kinds of Ebola vaccines were urgently under development across the world. Among these, the novel recombinant adenovirus type 5 vector-based Ebola vaccine (Ad5-EBOV)-the first Ebola vaccine based on the 2014 Zaire Guinea epidemic strain-was developed in China, and its safety and immunogenicity were demonstrated in China and Sierra Leone. The license to market the drug was approved on October 19, 2017, by the Chinese Food and Drug Administration. In order to standardize the test on the Ad5-EBOV virus titer, China's national standard substance for the virus titer of Ad5-EBOV was established according to the recommendations for the preparation, characterization, and establishment of international and other biological reference standards from the World Health Organization and Chinese Pharmacopoeia (third edition). The standard for the Ad5-EBOV virus titer was prepared with a volume of 0.5 mL per ampoule in lyophilized form. The samples of the standard, designated as A, B, C, D with different aims, were blinded and distributed to five laboratories to be collaboratively calibrated. The virus titer for this standard was determined with the antibody staining method according to the instructions in the Adeno-X™ Rapid Titer Kit. The homogeneity and stability of the standard substance were also satisfied. The virus titer standard value was 8.54 lg infectious units (IFU)/mL, and the 95% confidence interval was between 7.94 lg IFU/mL and 9.14 lg IFU/mL. This standard was approved by the Chinese national committee and is available on the National Institutes for Food and Drug Control Web site ( www.nifdc.org.cn ; lot no. 250019-201501).

Bioassay of recombinant protein products is important tests to ensure protein effectiveness. Some recombinant protein products have no cells used in their bioassay but instead use animal models, while others have no suitable method. Here, we developed a method to obtain responsive cells used in bioassay of proteins. After screening of a CRISPR/Cas9 library, we obtained a responsive cell line that grew faster in the presence of rhEGF (recombinant human epidermal growth factor) than that of control cells. We used this cell line for bioassay of rhEGF. This cell line, compared with the control cells, had a 2 day shorter operation time and had lower interference. The responsive cell line is more suitable for use in bioassay of rhEGF.

Rift Valley fever virus (RVFV) is a mosquito-borne pathogen that causes substantial morbidity and mortality in livestock and humans. To date, there are no licensed human vaccines or therapeutics available. Here, we report the isolation of monoclonal antibodies from a convalescent patient, targeting the RVFV envelope proteins Gn and Gc. The Gn-specific monoclonal antibodies exhibited much higher neutralizing activities in vitro and protection efficacies in mice against RVFV infection, compared to the Gc-specific monoclonal antibodies. The Gn monoclonal antibodies were found to interfere with soluble Gn binding to cells and prevent infection by blocking the attachment of virions to host cells. Structural analysis of Gn complexed with four Gn-specific monoclonal antibodies resulted in the definition of three antigenic patches (A, B and C) on Gn domain I. Both patches A and B are major neutralizing epitopes. Our results highlight the potential of antibody-based therapeutics and provide a structure-based rationale for designing vaccines against RVFV.

Background Pathogenic Gram-negative bacteria interact with their eukaryotic hosts by deploying the type III translocon to inject effector proteins into the cytosol of eukaryotic cells. The translocon compositions, the number and biochemical characteristics of type III translocators in animal-pathogenic bacteria have been well elucidated, but information is lacking for plant-pathogenic bacteria. With extensive studies on biological functions of the Hpa1 protein secreted by the type III secretion system in Xanthomonas oryzae pv. oryzae (Xoo), we show here that Hpa1 is a type III translocator based on measurements of two proteins categorized as transcription activator-like (TAL) effector. Results Hpa1 was functionally associated with the TAL effector PthXo1 or AvrXa10 by genetic analysis of the wild-type Xoo strain and related mutants or recombinant strains. Inoculation experiments suggested that Hpa1 is required not only for the virulent role of PthXo1 in the susceptible rice variety Nipponbare, but also for the avirulent function of AvrXa10 on the resistant rice variety IRBB10. Hpa1 is unrelated to the secretion of PthXo1 and AvrXa10 out of bacterial cells. However, Hpa1 is critical for both TAL effectors to be translocated from bacterial cells into the cytosol of rice cells based on replicate experiments performed on the susceptible and resistant varieties, respectively. Hpa1-mediated translocation of PthXo1 is coincident with induced expression of rice SWEET11 gene, which is the regulatory target of PthXo1, resulting in the occurrence of the bacterial blight disease in the susceptible rice variety. By contrast, the immune hypersensitive response is induced in agreement with induced expression of rice Xa10 gene, which is the target of AvrXa10, only when AvrXa10 is translocated from bacteria into cells of the resistant rice variety. All the virulent or avirulent performances of the TAL effectors are nullified by directed mutation that removes the α-helix motif from the Hpa1 sequence. Conclusions The genetic and biochemical data demonstrate that Hap1 is a type III translocator at least for TAL effectors PthXo1 and AvrXa10. The effect of the directed mutation suggests that Hpa1 depends on its α-helical motif to fulfil the translocator function. Electronic supplementary material The online version of this article (10.1186/s12866-018-1251-3) contains supplementary material, which is available to authorized users.

Development of anti-VEGF based biologic agents has been a focus in cancer treatment for the past decades, and several anti-VEGF pharmaceuticals have been already approved for treatment of various medical indications especially in cancer. The first anti-angiogenic agent approved by FDA was bevacizumab (BVZ, trade name Avastin, Genentech/Roche), a humanized anti-VEGF monoclonal antibody. Accurate determination of bioactivity is crucial for the safety and efficacy of therapeutic antibodies. The current method widely used in the lot release and stability test for clinical trial batches of BVZ is anti-proliferation assay using primary human umbilical vein endothelial cells (HUVEC), which is tedious with high assay variations. We describe here the development and preliminary validation of a reporter gene assay (RGA) that is based on an HEK293 cell line stably expressing vascular endothelial growth factor receptor 2 (VEGFR-2), and a luciferase reporter under the control of nuclear factor activated T cell (NFAT) response elements. Our study shows this assay not only to be superior on precision, sensitivity and assay simplicity compared with HUVEC assay, but also applicable to other VEGF-targeted biotherapeutics. These results show for the first time that this new reporter assay, based on the VEGF-VEGFR-NFAT pathway, can be a viable supplement to the HUVEC assay and employed in potency determination of BVZ and other kinds of anti-VEGF antibody-based biotherapeutics.

Previous observations have shown that the distribution of 36.2-GHz class I methanol megamaser (MM) emission in Arp 220 is highly correlated with the diffuse X-rays. On this basis it was suggested that methanol MM may be produced either by the effects of galactic-outflow-driven shocks and/or cosmic rays. Here we report the results of a single-dish survey undertaken with the Greenbank Telescope (GBT) to improve our understanding of the pumping conditions of extragalactic class I methanol masers and their relationship to starburst and feedback processes within the host galaxies, towards a sample which includes 16 galaxies which show both extended soft X-ray emission, and either OH or H2O MM emission. Large baseline ripples in the GBT spectra limited our results to tentative detections towards 11 of the target galaxies. Analysis of these tentative detections shows that there are significant correlations between the methanol intensity and the host-galaxy infrared, radio and OH MM emission, but no correlation with the X-ray and H2O MM emission. Some sources show methanol emission significantly offset from the systemic velocity of the galaxy (by up to 1000 km s−1) and we propose that these are associated with galactic-scale outflows from active galactic nuclei (AGNs) feedback. The combined observational properties suggest that class I methanol MMs are related to significant starburst and molecular outflow activity and hence may provide a potential probe of AGN feedback and starburst processes in the host galaxies.

We present observations of isotopologue lines of dense-gas tracers at 3 mm and 1 mm towards the nearest ultra-luminous infrared galaxy Arp 220. The 3-mm and 1-mm observations were performed with the Institut de Radioastronomie Millimetrique 30-m telescope and the Atacama Pathfinder Experiment 12-m telescope, respectively. We detected (HCN)-C-13 and (HNC)-C-13 in 1-0 and 3-2, and (HCN)-N-15 1-0, among which (HCN)-N-15 1-0 and (HNC)-C-13 1-0 are detected in Arp 220 for the first time. The (HCO+)-C-13 1-0 and 3-2 lines are unlikely to be detected because of the confusion of SiO lines. We find that the ratio of the line brightness temperatures of (HNC)-C-13 3-2/1-0 is 2.4, which is significantly higher than that of (HCN)-C-13 3-2/1-0 (0.73). This indicates that (HNC)-C-13 and HNC molecules are in denser regions than (HCN)-C-13 and HCN molecules. With the line ratio of (HCN)-C-13 1-0 and (HCN)-N-15 1-0, the N-14/N-15 ratio was estimated to be 440(-82)(+140), which is larger than that of the local interstellar medium.

We present Atacama Large Millimeter/submillimeter Array (ALMA) and Karl G. Jansky Very Large Array (JVLA) observations of the massive infrared dark cloud NGC 6334S (also known as IRDC G350.56+0.44), located at the southwestern end of the NGC 6334 molecular cloud complex. The H$^{13}$CO$^{+}$ and the NH$_{2}$D lines covered by the ALMA observations at a $\sim$3$^{\prime\prime}$ angular resolution ($\sim$0.02 pc) reveal that the spatially unresolved non-thermal motions are predominantly subsonic and transonic, a condition analogous to that found in low-mass star-forming molecular clouds. The observed supersonic non-thermal velocity dispersions in massive star forming regions, often reported in the literature, might be significantly biased by poor spatial resolutions that broaden the observed line widths due to unresolved motions within the telescope beam. Our 3~mm continuum image resolves 49 dense cores, whose masses range from 0.17 to 14 $M_{\odot}$. The majority of them are resolved with multiple velocity components. Our analyses of these gas velocity components find an anti-correlation between the gas mass and the virial parameter. This implies that the more massive structures tend to be more gravitationally unstable. Finally, we find that the external pressure in the NGC 6334S cloud is important in confining these dense structures, and may play a role in the formation of dense cores, and subsequently, the embedded young stars., 30 pages, 10 figures, 2 tables, accepted for publication in The Astrophysical Journal

We performed a radio recombination line (RRL) survey to construct a high-mass star-forming region (HMSFR) sample in the Milky Way based on the all-sky Wide-Field Infrared Survey Explorer ($\textit{All-WISE}$) point source catalog. The survey was observed with the Shanghai 65m Tianma radio telescope (TMRT) covering 10 hydrogen RRL transitions ranging from H98$\alpha$ to H113$\alpha$ (corresponding to the rest frequencies of 4.5$-$6.9 GHz) simultaneously. Out of 3348 selected targets, we identified an HMSFR sample consisting of 517 sources traced by RRLs, a large fraction of this sample (486) locate near the Galactic plane ($|$$\textit{b}$$|$ $, Comment: 24 pages, 19 figures

Metals are thought to have profound effects on the internal structures of molecular clouds in which stars are born. The absence of metals is expected to prevent gas from efficient cooling and fragmentation in theory. However, this effect has not yet been observed at low metallicity environments, such as in the early Universe and local dwarf galaxies, because of the lack of high spatial resolution maps of gas. We carried out ALMA observations of the carbon monoxide (CO) J=2-1 emission line at 1.4-parsec resolutions of a molecular cloud in DDO 70 at 7% solar metallicity, the most metal-poor galaxy currently known with a CO detection. In total, five clumps have been identified and they are found to follow more or less the Larson's law. Since the CO emission exists in regions with visual extinction A_V around 1.0, we converted this A_V to the gas mass surface density using a gas-to-dust ratio of 4,594+-2,848 for DDO 70. We found that the CO clumps in DDO 70 exhibit significantly larger (on average four times) sizes than those at the same gas mass surface densities in massive star-formation regions of the Milky Way. The existence of such large clumps appears to be consistent with theoretical expectations that gas fragmentation in low metallicity clouds is suppressed. While our observation is only for one cloud in the galaxy, if it is representative, the above result implies suppressed gas fragmentation during the cloud collapse and star formation in the early Universe., Comment: ApJ in press, 8 figures

Complex structure and structure-function relationship of biopharmaceuticals require extensive analytical characterization and appropriate quality control of the products. Despite rapid development of sophisticated physicochemical techniques, biological activity measurement remains the critical role in inferring the high-order structure of biopharmaceuticals. Cell-based biological assays are mostly applied to determine the biological activity of biopharmaceuticals, however, refined biological assays are continually needed to increase their robustness. Reporter gene assays (RGAs) which are mechanism of action (MOA) related, less variable, accurate, precise, and labor-saving are becoming more and more recognized and adopted in the quality control. Here we discuss the importance of bioactivity determination, the strength and weakness of various assay formats with RGAs. We also introduce the mechanism of RGAs, and present a number of examples for RGAs to determine the bioactivity of various biopharmaceuticals, which indicate their extensive use in the screening, characterization, quality control, stability and biosimilarity study. We believe that with the rapid development of biotechnology, new strategies of bioassays based on RGAs will be more widely applied in various fields of biopharmaceuticals.

We have performed a multi-wavelength study toward a quasi-sinusoidal filament (CFG028.68–0.28). A new large-scale 12CO J = 3 – 2 map was obtained from the China-Cologne Observation for SubMillimeter Astronomy (CCOSMA) 3m radio telescope. Based on the ATLASGAL catalog, we have identified 27 dust clumps in the filament. Through the relationship between the mass and radius of these clumps, 67% of these clumps are dense and massive enough to potentially form massive stars. The obtained CFE is ∼11% in the filament. The filament has a linear mass density of ∼305 M ⊙ pc−1, which is smaller than its critical mass to length ratio. This suggests that the external pressure from the neighboring H ii regions may help prevent the filament from dispersing under the effects of turbulence. Comparing the energy injection from outflows and H ii regions in the filament, the ionization feedback from the H ii regions can help maintain the observed turbulence.

The aim of this study was to elucidate the gastroprotective activity and possible mechanism of involvement of araloside A (ARA) against ethanol- and aspirin-induced gastric ulcer in mice. The experimental mice were randomly divided into control, model, omeprazole (20 mg/kg, orally) and ARA (10, 20 and 40 mg/kg, orally). Gastric ulcer in mice was induced by intragastric administration of 80% ethanol (10 mL/kg) containing 15 mg/mL aspirin 4 h after drug administration on day 7. The results indicated that ARA could significantly raise gastric juice volume and acidity; ameliorate gastric mucosal blood flow, gastric binding mucus volume, ulcer index and ulcer inhibition rate; suppress H

Soluble glycoprotein 130 kDa (sgp130)-Fc fusion protein, an innovative therapeutic bio-macromolecular drug specifically targeting IL-6 trans-signaling, proved to have good potential for application in the treatment of chronic inflammatory diseases. A simple and quick bioassay for sgp130-Fc was developed in this study. First, a stable reporter cell line was obtained by transfecting CHO-K1 cells with a sis-inducible element (SIE)-driving luciferase reporter gene (CHO/SIE-Luc). Sgp130-Fc could inhibit the expression of luciferase induced by IL-6/sIL-6R&alpha, complex, and the dose&ndash, response curve fitted the four-parameter logistic model, with 50% inhibitive concentration (IC50) being about 500 ng/mL and detection range between 40 and 5000 ng/mL. Both the intra-assay and inter-assay coefficient of variation (CV) were below 10.0%, and the accuracy estimates ranged from 94.1% to 106.2%. The assay indicated a good linearity (R&sup2, = 0.99) in the range of 50% to 150% of optimized initial concentration. No significant difference was found between the test results of new assay and BAF3/gp130 proliferation assay (unpaired t test, p = 0.4960, n = 6). The dose-response effect and copy number of the luciferase gene was basically unchanged after long-term culture (up to passage 60), demonstrating the stability of CHO/SIE-Luc cells. These results suggested that the new reporter assay was suited to routine potency determination of therapeutic sgp130-Fc.

We present HCN J=4-3 and HCO^+ J=4-3 maps of six nearby star-forming galaxies, NGC 253, NGC 1068, IC 342, M82, M83, and NGC 6946, obtained with the James Clerk Maxwell Telescope as part of the MALATANG survey. All galaxies were mapped in the central 2 arcmin $\times$ 2 arcmin region at 14 arcsec (FWHM) resolution (corresponding to linear scales of ~ 0.2-1.0 kpc). The L_IR-L'_dense relation, where the dense gas is traced by the HCN J=4-3 and the HCO^+ J=4-3 emission, measured in our sample of spatially-resolved galaxies is found to follow the linear correlation established globally in galaxies within the scatter. We find that the luminosity ratio, L_IR/L'_dense, shows systematic variations with L_IR within individual spatially resolved galaxies, whereas the galaxy-integrated ratios vary little. A rising trend is also found between L_IR/L'_dense ratio and the warm-dust temperature gauged by the 70 \mu m/100 \mu m flux ratio. We find the luminosity ratios of IR/HCN(4-3) and IR/HCO^+(4-3), which can be taken as a proxy for the efficiency of star formation in the dense molecular gas (SFE_dense), appears to be nearly independent of the dense-gas fraction (f_dense) for our sample of galaxies. The SFE of the total molecular gas (SFE_mol) is found to increase substantially with f_dense when combining our data with that on local (ultra)luminous infrared galaxies and high-z quasars. The mean L'_HCN(4-3)/L'_HCO^+(4-3) line ratio measured for the six targeted galaxies is 0.9+/-0.6. No significant correlation is found for the L'_HCN(4-3)/L'_HCO^+(4-3) ratio with the SFR as traced by L_IR, nor with the warm-dust temperature, for the different populations of galaxies., Comment: 29 pages, 12 figures, accepted for publication in ApJ

Outbreaks of the highly pathogenic avian influenza H5N1 and H7N9 viruses have spurred an unprecedented research effort to develop antivirals and vaccines against influenza. Standardized methods for vaccine evaluation are critical for facilitating vaccine development. Compared with hemagglutination inhibition assays, mounting evidence suggest that microneutralization tests (MNTs) is a better choice for the evaluation of candidate pandemic influenza vaccines because they measure neutralizing antibody activity in cell cultures and are more sensitive in detecting H5 and H7. Here, we report a MNT measuring neuraminidase activity as the read-out (NA-MNT) for quantitative analysis of neutralizing antibodies against avian influenza viruses. Compared to the conventional microneutralization assay (ELISA-MNT), the NA-MNT is faster with lower intra- and inter-assay variations, while no difference in geometric mean titers was found between these two assays for the evaluation of H5N1 and H7N9 vaccines. These results suggest that NA-MNT is a reliable and high throughput method which could facilitate the development of candidate pandemic influenza vaccine.

IL-6 has an important role in the pathogenesis of autoimmunity and chronic inflammation. Several mAbs that target IL-6 or the IL-6 receptor (IL-6R) have been established and approved for the treatment of various diseases such as multicentric Castleman's disease and rheumatoid arthritis. Quality control of therapeutic antibodies requires accurate determination of bioactivity. However, current cell-based anti-proliferation assays are tedious, time consuming, and result in high variation. We therefore developed a reporter gene assay (RGA) based on an IL-6-dependent DS-1 cell line that stably expressed the reporter luciferase controlled by the serum-induced element (SIE) response element, which was a key element located downstream of the IL-6 signaling pathway. The RGA method demonstrated good performance characteristics after careful optimization, including high specificity, stability, accuracy, precision, and robustness. It also had superior precision and sensitivity. The assay is simple compared with the traditional anti-proliferation assay. This novel RGA based on the IL-6-IL-6R-STAT3 pathway can be useful, in conjunction with the anti-proliferation bioassay, to determine the bioactivity of anti-IL-6/anti-IL-6R therapeutic mAbs. Graphical abstract The mechanism sketch of the reporter gene assay for the bioactivity determination of anti-IL-6/anti-IL-6Rα mAbs.

MicroRNAs (miRNAs) have been reported to be associated with heart valve disease, which can be caused by inflammation. This study aimed to investigate the functional impacts of miR-27a on TNF-α-induced inflammatory injury in human mitral valve interstitial cells (hMVICs). hMVICs were subjected to 40 ng/mL TNF-α for 48 h, before which the expressions of miR-27a and NELL-1 in hMVICs were altered by stable transfection. Trypan blue staining, BrdU incorporation assay, flow cytometry detection, ELISA, and western blot assay were performed to detect cell proliferation, apoptosis, and the release of proinflammatory cytokines. We found that miR-27a was lowly expressed in response to TNF-α exposure in hMVICs. Overexpression of miR-27a rescued hMVICs from TNF-α-induced inflammatory injury, as cell viability and BrdU incorporation were increased, apoptotic cell rate was decreased, Bcl-2 was up-regulated, Bax and cleaved caspase-3/9 were down-regulated, and the release of IL-1β, IL-6, and MMP-9 were reduced. NELL-1 was positively regulated by miR-27a, and NELL-1 up-regulation exhibited protective functions during TNF-α-induced cell damage. Furthermore, miR-27a blocked JNK and Wnt/β-catenin signaling pathways, and the blockage was abolished when NELL-1 was silenced. This study demonstrated that miR-27a overexpression protected hMVICs from TNF-α-induced cell damage, which might be via up-regulation of NELL-1 and thus modulation of JNK and Wnt/β-catenin signaling pathways.

Both influenza viral hemagglutinin and neuraminidase can induce protective immune responses in humans. Although the viral hemagglutinin antigens have been quantified in influenza vaccines, the amounts of neuraminidase remain undetermined. Using comprehensive bioinformatics analyses of all neuraminidase sequences, we identified highly conserved and subtype-specific peptide epitopes within each of N1, N2 and type B neuraminidase groups. Mono-specific antibodies generated against these peptides bound to their respective subtype/type only while demonstrating remarkable specificity against the viral neuraminidase sequences without any cross-reactivity with allantoic and cellular proteins. Moreover, the subtype/type-specific antibodies were found not to interfere with one another when a mixture of vaccine samples was analysed. Importantly, immunoassay based on these antibodies can quantitatively determine neuraminidase in commercial trivalent vaccine samples. Analyses of vaccines from eight manufacturers using the same vaccine seeds revealed significant differences in neuraminidase levels. Specifically, while the ratio between neuraminidase and hemagglutinin in some products are found to be close 1/5, other products have a ratio of approximately 1/100, a level which is far below the theoretical ratio between neuraminidase and hemagglutinin in a virus. The antibody-based assays reported here could be of great value for better quality control of both monovalent and trivalent vaccines.

We present results of millimeter line observations toward four local gas-rich galaxies (NGC 3079, NGC 4258, NGC 6240 and VII Zw 31) with the IRAM 30 meter millimeter telescope. More than 33 lines in these four sources were detected, including normal dense gas tracers (HCN 1-0, HCO$^+$ 1-0, and C$_2$H 1-0, etc) and their isotopic species. H$^{13}$CN (1-0) and H$^{13}$CO$^+$ (1-0) are detected for the first time in NGC 4258. Optical depths of HCN 1-0 and HCO$^{+}$ 1-0 were estimated with detected isotopic lines in NGC 4258, which were 4.1 and 2.6, respectively. HC$_3$N $J=29-28$, which requires high volume density and high temperature to excite, was detected in NGC 6240. High ratios of HCO$^+$/HCN in NGC 4258 and NGC 6240 imply that this ratio might not be a perfect diagnostic tool between AGN and starburst environments, due to contamination/combination of both processes. The low HC$_3$N/HCN line ratios with less than 0.15 in NGC 4258, NGC 6240 and the non-detection of HC$_3$N line in NGC 3079 and VII Zw 31 indicates that these four galaxies are HC$_3$N-poor galaxies. The variation of fractional abundance of CN in different types of galaxies is large., Comment: 15pages, 13 figures; accepted for publication in MNRAS

This paper puts forward a alternative and direct current electric power iteration algorithm based on twice sample. The algorithm is aimed at non-sine signal model overlaid by DC and AC component and is used for measuring alternative and direct current power of specified-frequency electric signal by calculating the power value of each DC and AC component. By means of MATLAB simulation, the accuracy and rapidity of this algorithm is verified. And obtain the influence of bandwidth parameter on power tracking error. Adjusting the parameter can improve the rate of power estimation.

MRC-5 represents the most frequent human diploid cells (HDCs)-type cell substrate in the production of human viral vaccines. However, early-passage MRC-5 is diminishing and, due to both technical and ethical issues, it is extremely difficult to derive novel HDCs from fetal lung tissues, which are the common sources of HDCs. Our previous studies suggested that human umbilical cord may represent an alternative but convenient source of new HDCs. Here, we established a three-tiered cell banking system of a hUC-MSC line, designated previously as Cell Collection and Research Center-1 (CCRC-1). The full characterization indicated that the banked CCRC-1 cells were free from adventitious agents and remained non-tumorigenic. The CCRC-1 cells sustained its rapid proliferation even at passage 30 and were susceptible to the infection of a wide spectrum of viruses. Interestingly, the CCRC-1 cells showed much higher production of EV71 or Rubella viruses than MRC-5 and Vero cells when growing in serum-free medium. More importantly, the EV71 vaccine produced from CCRC-1 cells induced immunogenicity while eliciting no detectable toxicities in the tested mice. Collectively, these studies further supported that CCRC-1, and likely other hUC-MSCs as well, may serve as novel, safe and high-yielding HDCs for the production of human viral vaccines.

We report the detection of widespread CH$_2$OHCHO and HOCH$_2$CH$_2$OH emission in Galactic center giant molecular cloud Sagittarius B2 using the Shanghai Tianma 65m Radio Telescope. Our observations show for the first time that the spatial distribution of these two important prebiotic molecules extends over 15 arc-minutes, corresponding to a linear size of approximately 36 pc. These two molecules are not just distributed in or near the hot cores. The abundance of these two molecules seems to decrease from the cold outer region to the central region associated with star-formation activity. Results present here suggest that these two molecules are likely to form through a low temperature process. Recent theoretical and experimental studies demonstrated that prebiotic molecules can be efficiently formed in icy grain mantles through several pathways. However, these complex ice features cannot be directly observed, and most constraints on the ice compositions come from millimeter observations of desorbed ice chemistry products. These results, combined with laboratory studies, strongly support the existence of abundant prebiotic molecules in ices., 20 pages, 7 figures, accepted by ApJ

Several studies have reported the preventive or therapeutic effect of curcuminoids on chronic heart failure (CHF), but their application was limited due to low solubility and bioavailability. Our previous study indicates that self-emulsifying drug delivery system (SEDDS) improves the solubility and bioavailability of curcuminoids. Thus, the aim of this work was to investigate whether SEDDS could improve preventive effect of curcuminoids on CHF in rats. CHF model was were established by coronary artery ligation. Ninety rats were randomly and averagely divided into sham, model, low- or high-dose suspension or SEDDS of curcuminoids (66.68 or 266.68 mg/kg) groups. Hemodynamic indices were recorded by multipurpose polygraph. Serum oxidative indices, B-type natriuretic peptide (BNP) and heart weight index were determined by kits and electronic balance. Myocardial infarct area, ventricular dilatation degree and collagen volume fraction of myocardial interstitium were analyzed by Masson staining, picric acid and sirius red staining, light microscopy and image analysis system. Myocardial histopathology was observed by hematoxylin and eosin staining, Masson staining and light microscopy. Reduction of ventricular pump function, increase of BNP level and heart weight index, myocardial lipid peroxidation damage, myocardial infarction, myocardial fibrosis, and cardiac enlargement were detected or observed in model group relative to those in sham group. After treatment with suspension or SEDDS of curcuminoids, the above-mentioned pathological changes were obviously reversed relative to those in model group. Meanwhile, the ameliorative effect of SEDDS of curcuminoids was markedly better than that of suspension of curcuminoids. This work provides a valuable reference from pharmacodynamics for development of curcuminoids pharmaceutics. Keywords: Curcuminoids, Self-emulsifying drug delivery system, Chronic heart failure, Ventricular pump function, Lipid peroxidation damage

Bisabolangelone is the main active ingredient of Angelica polymorpha, which has the good pharmacological activities such as anti-celialgia, anti-dysmenorrhea and anti-gastralgia. In this study, the neuroprotective effect of bisabolangelone against hydrogen peroxide (H2O2)-induced neurotoxicity in human neuroblastoma cells was studied. The binding capacity between bisabolangelone and calcium channel was calculated by molecular docking method, and calcium concentration was detected by the calcium fluorescent probe of Fura-2 AM. The results indicated that bisabolangelone had a good neuroprotective effect on nerve cells by interacting with calcium channels, mainly blocking the voltage-gated calcium channels, suppressing calcium releasing from the calcium store.

The present study aimed to develop a symbiotic selection-marker-free plasmid and host system that would allow successful plasmid maintenance and amplification for use in gene therapy. Initially, the chromosomal aspartate‑semialdehyde dehydrogenase (asd) gene was disrupted in DH10B Escherichia coli using Red recombinase‑mediated homologous recombination. This method required the use of linear DNA fragments carrying kan‑kil genes, and/or homologous extensions to the targeted locus. The resultant auxotrophic cell wall‑deficient strain (DH10BΔasd) was evaluated as a symbiotic host for amplification of the marker‑free plasmid, allowing it to supply ASD activity. In order to construct the plasmid, an asd expression cassette was inserted, under the control of the nirB promoter, into a eukaryotic expression vector, and its kanamycin resistance gene was subsequently removed. The symbiotic plasmid and host system was assessed for numerous plasmid production and stability parameters, including structure, yield, plasmid‑retention rate, and bacterial storability, under various conditions. The presence of the plasmid was subsequently confirmed by growth test, restriction enzyme mapping, and sequencing. The plasmid yield and copy number produced in the symbiotic cells, in the absence of antibiotic selection, were shown to be similar to those produced under kanamycin selection, in the cells containing the precursor plasmid and kanamycin resistance gene. Furthermore, the results of the present study demonstrated that when inoculated with1% inoculant volume,98% of the cells in the culture retained the plasmid regardless of the number of passages. The strain was stable when stored at ‑70˚C, with negligible viability loss over 12 months. The constructed plasmid is stable and has potential in future gene therapy, while much work is still required.

Up to now, all tested Ebola virus vaccines have been based on the virus strain from the Zaire outbreak in 1976. We aimed to assess the safety and immunogenicity of a novel recombinant adenovirus type-5 vector-based Ebola vaccine expressing the glycoprotein of the 2014 epidemic strain.We did this randomised, double-blind, placebo-controlled, phase 1 clinical trial at one site in Taizhou County, Jiangsu Province, China. Healthy adults (aged 18-60 years) were sequentially enrolled and randomly assigned (2:1), by computer-generated block randomisation (block size of six), to receive placebo, low-dose adenovirus type-5 vector-based Ebola vaccine, or high-dose vaccine. Randomisation was pre-stratified by dose group. All participants, investigators, and laboratory staff were masked to treatment allocation. The primary safety endpoint was occurrence of solicited adverse reactions within 7 days of vaccination. The primary immunogenicity endpoints were glycoprotein-specific antibody titres and T-cell responses at day 28 after the vaccination. Analysis was by intention to treat. The study is registered with ClinicalTrials.gov, number NCT02326194.Between Dec 28, 2014, and Jan 9, 2015, 120 participants were enrolled and randomly assigned to receive placebo (n=40), low-dose vaccine (n=40), or high-dose vaccine. Participants were followed up for 28 days. Overall, 82 (68%) participants reported at least one solicited adverse reaction within 7 days of vaccination (n=19 in the placebo group vs n=27 in the low-dose group vs n=36 in the high-dose group; p=0·0002). The most common reaction was mild pain at the injection site, which was reported in eight (20%) participants in the placebo group, 14 (35%) participants in the low-dose group, and 29 (73%) participants in the high-dose vaccine group (p0·0001). We recorded no statistical differences in other adverse reactions and laboratory tests across groups. Glycoprotein-specific antibody titres were significantly increased in participants in the low-dose and high-dose vaccine groups at both day 14 (geometric mean titre 421·4 [95% CI 249·7-711·3] and 820·5 [598·9-1124·0], respectively; p0·0001) and day 28 (682·7 [424·3-1098·5] and 1305·7 [970·1-1757·2], respectively; p0·0001). T-cell responses peaked at day 14 at a median of 465·0 spot-forming cells (IQR 180·0-1202·5) in participants in the low-dose group and 765·0 cells (400·0-1460·0) in those in the high-dose group. 21 (18%) participants had mild fever (n=9 in the placebo group, n=6 in the low-dose group, and n=6 in the high-dose group). No serious adverse events were recorded.Our findings show that the high-dose vaccine is safe and robustly immunogenic. One shot of the high-dose vaccine could mount glycoprotein-specific humoral and T-cell response against Ebola virus in 14 days.China National Science and Technology, Beijing Institute of Biotechnology, and Tianjin CanSino Biotechnology.