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To investigate the effect of silencing DNA methyltransferase 1(DNMT1) to the methylation of the promoter of the tumor suppressor gene wnt-1 (WIF-1) in human chronic myeloid leukemia (CML) cells.DNMT1 siRNAi plasmid was constructed and DNMT1 siRNAi was transfected into CML K562 cells. RT-PCR and Western blot were used to detect the expression of DNMT1 gene and related protein, and methylation PCR was used to detect WIF-1 gene promoter methylation level. The trypan blue exclusion and MTT assay were used to detect the cell proliferation, flow cytometry were used to detect the cell apoptosis rate, colony formation assay was used to detect cell colony formation ability. Expression of Wnt/β- catenin and its downstream signaling pathway proteins were detected by Western blot after DNMT1 gene was silenced.The expression level of DNMT1 mRNA and its related protein in the experimental group were significantly lower than those in the control group and negative control group (P0.05). After 72 hours of successful transfection, the WIF-1 gene in the control group and negative control group were completely methylated, while in the experimental group, the methylation level significantly decreased. The results of MSP showed that the PCR product amplified by the unmethylated WIF-1 primer in the experimental group increased significantly,while by the methylated WIF-1 primer decreased significantly. After 48 h of transfection, the OD value, viable cell number and colony formation of the cells in experimental group were significantly lower than those in the negative control group and the control group (P0.05). The apoptosis rate of the cells in experimental group was significantly higher than those in the negative control group and control group (P0.05). The expression levels of β- actin, myc, cyclin D1 and TCF-1 in K562 cells in the experimental group were significantly lower than those in the negative control group and control group (P0.05).Silencing DNMT1 gene can inhibit the proliferation and promote the apoptosis of K562 cells. The mechanism may be related to reverse the hypermethylation level of the WIF-1 gene promoter, thereby inhibit the activity of the Wnt/β- catenin signaling pathway.沉默DNMT1减弱WIF-1基因启动子甲基化对慢性髓系白血病K562细胞生物学行为的影响.探讨沉默DNA甲基转移酶（DNMT1）对人慢性髓系白血病细胞内抑癌基因wnt抑制因子-1（WIF-1）基因启动子甲基化的影响。.构建DNMT1 siRNAi质粒，将DNMT1 siRNAi转染至慢性髓系白血病K562细胞中，采用RT-PCR和Western blot法检测DNMT1基因和相关蛋白的表达，并利用甲基化PCR检测WIF-1基因启动子甲基化水平，台盼蓝拒染法和MTT 法检测细胞的增殖能力，流式细胞术检测细胞的凋亡率，集落形成实验检测细胞的集落形成能力，Western blot法检测沉默DNMT1基因后 Wnt/β-catenin及其下游信号通路蛋白的表达情况。.实验组DNMT1 mRNA和相关蛋白表达水平显著低于对照组和阴性对照组（P0.05）；成功转染72 h后，对照组和阴性对照组WIF-1基因处于完全甲基化状态，而实验组WIF-1基因启动子区域甲基化水平显著降低。MSP检测结果显示，实验组未甲基化的WIF-1引物扩增出的PCR产物含量显著增加，而甲基化的WIF-1引物扩增出的PCR产物含量显著降低。转染成功48 h后，实验组的OD值、活细胞数及集落形成数均显著低于阴性对照组和对照组（P0.05）。实验组的细胞凋亡率显著高于阴性对照组和对照组（P0.05）。实验组K562细胞中β-actin、myc、cyclin D1和TCF-1蛋白表达水平均显著低于阴性对照组和对照组（P0.05。.沉默DNMT1基因能够抑制 K562 细胞增殖，促进其凋亡，其作用机制可能与逆转WIF-1基因启动子高甲基化水平，从而抑制Wnt/β-catenin信号通路的活性有关。.

Multiple myeloma (MM) is one of the most common tumors of the hematological system and remains incurable. Recent studies have shown that long noncoding RNA NORAD is a potential oncogene in a variety of tumors. However, the general biological role and clinical value of NORAD in MM remains unknown. In this study, we measured NORAD expression in bone marrow of 60 newly diagnosed MM, 30 post treatment MM and 17 healthy donors by real-time quantitative polymerase chain reaction (qPCR). The NORAD gene was knockdown by lentiviral transfection in MM cell lines, and the effects of NORAD on apoptosis, cell cycle and cell proliferation in MM cells were examined by flow cytometry, CCK8 assay, EDU assay and Western blot, and the differential genes after knockdown of NORAD were screened by mRNA sequencing, followed by in vivo experiments and immunohistochemical assays. We found that knockdown of NORAD promoted MM cell apoptosis, induced cell cycle G1 phase arrest, and inhibited MM cell apoptosis in in vivo and in vitro experiments. Mechanistically, NORAD plays these roles through the BMP6/P-ERK1/2 axis. We discuss a novel mechanism by which NORAD acts pro-tumorigenically in MM via the BMP6/P-ERK1/2 axis.

Natural killer (NK) cells preferentially accumulate at maternal–foetal interface and are believed to play vital immune‐modulatory roles during early pregnancy and related immunological dysfunction may result in pregnant failure such as recurrent miscarriage (RM). However, the mechanisms underlying the establishment of maternal–foetal immunotolerance are complex but clarifying the roles of decidual NK (dNK) cells offers the potential to design immunotherapeutic strategies to assist RM patients. In this report, we analysed RNA sequencing on peripheral NK (pNK) and decidual NK cells during early pregnancy; we identified an immunomodulatory dNK subset CXCR4+CD56brightdNK and investigated its origin and phenotypic and functional characteristics. CXCR4+CD56brightdNK displayed a less activated and cytotoxic phenotype but an enhanced immunomodulatory potential relative to the CXCR4 negative subset. CXCR4+CD56brightdNK promote Th2 shift in an IL‐4‐dependent manner and can be recruited from peripheral blood and reprogramed by trophoblasts, as an active participant in the establishment of immune‐tolerance during early pregnancy. Diminished CXCR4+ dNK cells and their impaired ability to induce Th2 differentiation were found in RM patients and mouse models of spontaneous abortion. Moreover, adoptive transfer of CXCR4+ dNK cells to NK‐deficient (Nfil3–/–) mice showed great therapeutic potential of CXCR4+ dNK via recovering the Th2/Th1 bias and reducing embryo resorption rates. The identification of this new dNK cell subset may lay the foundation for understanding NK cell mechanisms in early pregnancy and provide potential prognostic factors for the diagnosis and therapy of RM.

Chen Yuan,1,* Yanhong Li,1,* Liyun Liu,1 Baihetiya Tayier,1 Lingjie Yang,1 Lina Guan,1– 3 Yuming Mu1– 3 1Department of Echocardiography, First Affiliated Hospital of Xinjiang Medical University, Urumqi, 830011, People’s Republic of China; 2State Key Laboratory of Pathogenesis, Prevention and Treatment of High Incidence Diseases in Central Asia, First Affiliated Hospital of Xinjiang Medical University, Urumqi, People’s Republic of China; 3Xinjiang Key Laboratory of Medical Animal Model Research, Clinical Medical Research Institute of First Affiliated Hospital of Xinjiang Medical University, Urumqi, People’s Republic of China*These authors contributed equally to this workCorrespondence: Yuming Mu; Lina GuanDepartment of Echocardiography, The First Affiliated Hospital of Xinjiang Medical University, 137 Liyushan Road, Ürümqi, Xinjiang Uyghur, Autonomous Region, 830054, People’s Republic of ChinaEmail mym1234@126.com; sanjin_lsx@163.comObjective: To prepare a new type of dual-target microbubble loaded with anti-miR-33 (ANM33).Methods: Carrier core nanobubbles (NBs) were prepared by thin film hydration, and microbubbles loaded with PM1 (PCNBs) were prepared by grafting DSPE-PEG2000-maleimide-PM1 onto the NB surface. ANM33 was connected via electrostatic adsorption and covalent bonding, and hyaluronic acid (HA) was covalently connected. PM1 and HA were the targets, and ANM33 was the intervention drug. To evaluate the general physical and chemical properties of the prepared dual-target microbubbles loaded with ANM33 (HA-PANBs), we observed their morphology, particle size and surface potential while monitoring their stability and in vitro imaging ability, evaluated their toxic effect on cells and verified their ability to target cells.Results: HA-PANBs had a regular morphology and good stability. The average particle size measured by a Malvern potentiometer was 1421.75± 163.23 nm, and the average surface potential was − 5.51± 1.87 mV. PM1 and ANM33 were effectively connected to the NBs. The PM1, ANM33, and HA binding reached 89.0± 1.1%, 65.02± 5.0%, and 61.4± 3.5%, respectively, and the maximum binding reached 2 μg, 5 μg, and 7 μg/108 microbubbles, respectively. HA-PANBs had no obvious toxic effects on cells, and their ability to continuously enhance imaging in vitro persisted for more than 15 minutes, obviously targeting foam cells in the early stage of AS.Conclusion: HA-PANBs are ideal ultrasound contrast agents. The successful, firm connection of PM1 and HA to the NBs significantly increased the amount of carried ANM33. When microbubbles prepared with 2:4:7 PM1:ANM33:HA were used as a contrast agent, they had a high ANM33 carrying capacity, stable physical properties, and significantly enhanced imaging and targeting of foam cells in the early stage of AS.Keywords: ultrasound microbubbles, molecular targeting, dual targeting, atherosclerosis

The efficient triggering of prodrug release has become a challengeable task for stimuli-responsive nanomedicine utilized in cancer therapy due to the subtle differences between normal and tumor tissues and heterogeneity. In this work, a dual ROS-responsive nanocarriers with the ability to self-regulate the ROS level was constructed, which could gradually respond to the endogenous ROS to achieve effective, hierarchical and specific drug release in cancer cells. In brief, DOX was conjugated with MSNs via thioketal bonds and loaded with β-Lapachone. TPP modified chitosan was then coated to fabricate nanocarriers for mitochondria-specific delivery. The resultant nanocarriers respond to the endogenous ROS and release Lap specifically in cancer cells. Subsequently, the released Lap self-regulated the ROS level, resulting in the specific DOX release and mitochondrial damage in situ, enhancing synergistic oxidation-chemotherapy. The tumor inhibition Ratio was achieved to 78.49%. The multi-functional platform provides a novel remote drug delivery system in vivo.

We report that phosphorane can activate (salen)TiCl2 complex to achieve unprecedented excellent enantioselectivity and a broad substrate scope in the cyanation of nitroolefins. Our cyanating reagent Me2(CH2Cl)SiCN proves to be more active than TMSCN in this reaction, allowing 11 β-aliphatic nitrolefins and 12 β-CF3 nitroolefins (either β-aryl or aliphatic) to work well to give the corresponding tertiary or quaternary β-nitronitriles with high to excellent enantioselectivity.

Mg doping effects on the dielectric response and relaxation behavior in Ba2Co2-xMgxFe12O22 are experimentally studied for doping range of 0 ≤ x ≤ 1 through measurements of dielectric constant, loss and impedance spectroscopy as functions of temperature and frequency. It is shown that a colossal dielectric response with dielectric constant higher than 103 near room temperature for a wide range of frequency up to ~1 MHz can be realized by Mg doping at Co sites. Our results demonstrate that Mg doping can cause three obvious effects, namely, (i) to decrease Co3+ ions and in turn increase Fe ions in octahedrons, (ii) to raise the activation energy of relaxation and (iii) to increase resistance and capacitance of grain boundary. Our results also demonstrate that the dielectric relaxation is gradually changed from the intragranular- to the intergranular-dominated relaxation with increasing Mg doping content, perhaps due to the doping-caused increase in both resistance and capacitance of grain boundary.

Three phenylpropanoid glucosides (1 – 3) and one iridoid glucoside (11), together with eleven known glucosides, were isolated from the ethanol extract of the whole plant of Hemiphragma heterophyllum. Their structures were elucidated by means of 1D and 2D NMR spectroscopy, HRMS, and chemical methods. All compounds except 11 and 13 – 15 showed varying degrees of α-glucosidase inhibitory activity. Compounds 5, 9, and 12 were marginally active in the bioassay, while compounds 1 – 4, 6 – 8, and 10 exhibited appreciable inhibitory activity with an IC50 value of 33.6 ~ 83.1 µM, which was much lower than that of the positive control acarbose (IC50 = 310.8 µM).

Macrophage migration inhibitory factor (MIF) has been recognized as a major player in the pathogenesis of atherosclerosis. This study determined the association between polymorphisms of MIF gene and acute coronary syndrome (ACS). The polymorphism of MIF gene (rs755622, rs1007888 and rs2096525) was analyzed in 1153 healthy controls and 699 ACS cases in Chinese Han population. Plasma MIF level was also measured in part of ACS patients (139/19.9%) and healthy controls (129/11.2%) randomly. Most participants including healthy controls and ACS patients carried rs755622 GG (63.1% vs. 56.7%) and CG genotypes (33.1% vs. 38.9%) and G allele of rs755622 (79.6% vs. 76.1%, respectively), while CC genotype (3.8% vs. 4.4%) and C allele (20.4% vs. 23.9%) carriers were the lowest. Multivariate logistic regression analysis showed that carriers with rs755622 C allele had a higher risk of ACS compared to other genotypes (AOR = 1.278, 95% CI: 1.042–1.567). In addition, CC genotype carriers had the highest plasma levels of MIF than other genotype carriers. The MIF level in ACS patients with CC genotype was significantly higher than ACS patients carrying GG genotype and healthy controls carrying 3 different genotypes of MIF gene rs755622. Our findings indicate that MIF gene rs755622 variant C allele is associated with increased risk of ACS. Identification of this MIF gene polymorphism may help for predicting the risk of ACS.

Poly(amidoamine) (PAMAM) dendrimers are widely used as templates for synthesizing small nanoparticles. Herein, the surfaces of amine-terminated generation 4 (G4-NH2) poly(amidoamine) (PAMAM) dendri...

Guang-Lei Qiao,*,1 Li Chen,*,2 Wei-Hua Jiang,*,1 Cheng Yang,3 Chun-Mei Yang,4 Li-Na Song,1 Ying Chen,1 Hong-Li Yan,5 Li-Jun Ma11Department of Oncology, Tongren Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, People’s Republic of China; 2Department of Emergency Medicine, Renji Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai, People’s Republic of China; 3Department of Special Treatment, Third Affiliated Hospital of Second Military Medical University, Shanghai, People’s Republic of China; 4Department of Clinical Laboratory Diagnostics, Bei Hua University School, Jilin, People’s Republic of China; 5Department of Laboratory Diagnosis, and Reproductive Medicine Center, Changhai Hospital, Second Military Medical University, Shanghai, People’s Republic of China*These authors contributed equally to this workBackground: Circular RNAs (circRNAs) play important roles in the progression of cancers, but the precise role of circRNAs in the diagnosis and prognosis of hepatocellular carcinoma (HCC) remains to be clarified. The aim of the current study was to explore the diagnostic and prognostic values of hsa_circ_0003998 in HCC.Methods: CircRNAs expression was measured using RNA-seq analysis from HCC tissues (n=6) (three cases with or without portal vein invasion). Hsa_circ_0003998 in 200 pairs of HCC and adjacent noncancerous tissues and HCC cell lines was examined using qRT-PCR and the clinicopathologic significance was determined. We also detected the plasma levels of hsa_circ_0003998 in HCC, hepatitis B patients and healthy controls. The clinical diagnosis and prognostic values were further determined using receiver operating characteristic (ROC) curve, Kaplan–Meier curve and Cox regression.Results: Hsa_circ_0003998 was upregulated in HCC tissues (P

The activities and gene expression of antioxidative enzymes and the ROS content were analyzed in two typical storage-tolerant cultivars (Xushu 32 and Shangshu 19) and another two storage-sensitive cultivars (Yanshu 25 and Sushu 16) to explore the association between the storage capacity of sweetpotato (Ipomoea batatas (L.) Lam) and ROS scavenging capability. The storage roots of the storage-tolerant cultivars maintained higher activities and expression levels of antioxidative enzymes, including ascorbate peroxidase (APX), peroxidase (POD), catalase (CAT), and superoxide dismutase (SOD); lower activity and expression of lipoxygenase (LOX); and lower accumulation of ROS metabolites compared with the storage-sensitive cultivars. The antioxidative capability and ROS parameters of leaves were positively correlated with those of storage roots. Our results provide valuable insight for evaluating the storability of sweetpotato cultivars by analyzing the capabilities of the antioxidative system and the contents of ROS metabolites.

Nanoradiosensitizers are promising agents for enhancing cancer radiotherapeutic efficiency. Although many attempts have been adopted to improve their radiation enhancement effect through regulation of their size, shape, and/or surface chemistry, few methods have achieved satisfactory radiotherapeutic outcomes. Herein, we propose a sequential drug treatment strategy through cell cycle regulation for achieving improved radiotherapeutic performance of the nanoradiosensitizers. Docetaxel (DTX), a clinically approved first-line drug in breast cancer treatment, is first used to affect the cell cycle distribution and arrest cells in the G2/M phase, which has been proven to be the most effective phase for endocytosis and the most radiosensitive phase for radiotherapy. The cells are then exposed to a commonly used nanoradiosensitizer, gold nanoparticles (GNPs), followed by X-ray irradiation. It is found that by arresting the cancer cells in G2/M phase via the DTX pretreatment, the cellular internalization of GNPs is significantly promoted, therefore enhancing the radiosensitivity of cancer cells. The sensitization enhancement ratio of this sequential DTX/GNP treatment reaches 1.91, which is significantly higher than that (1.29) of GNP treatment. Considering its low cost, simple design, and high feasibility, this sequential drug delivery strategy may hold great potential in radiotherapy.

Aspergillus niger, which is a fungal pathogen, causes rot in a variety of fruits. In this study, the cystathionine β-synthase cbsA gene was deleted by homologous recombination to study its role in sulfur metabolism and pathogenicity of A. niger. The results showed that Δ cbsA strain maintained normal mycelia growth and sporulation compared with the control strain A. niger MA 70.15, whereas the contents of cysteine and glutathione (GSH) increased significantly after cbsA deletion. However, Δ cbsA strain showed reduced endogenous H2S production. Further results showed that cbsA gene deletion induced higher resistance to cadmium stress and stronger infectivity to pears. It was also found that a stronger response of reactive oxygen species (ROS) production was induced in Δ cbsA mutant-infected pear compared with the control strain. In all, the present research suggested the important role of cbsA in sulfur metabolism and pathogenicity of A. niger in pear fruit.

Assessment of the bacterial diversity associated with a decaying fern, Athyrium wallichianum Ching, revealed the presence of a novel bacterial strain named M46T. It was Gram-stain-negative, rod-shaped, non-motile and aerobic with cellulose and xylan degradation abilities. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain M46T was affiliated to the genus Sphingobacterium , exhibiting the highest sequence similarity of 97.9 % to Sphingobacterium ginsenosidimutans THG 07T, Sphingobacterium canadense CR11T and Sphingobacterium detergens 6.2 ST. Multilocus sequence analysis (MLSA) based on concatenated sequences of the rpoB, cpn60 and 16S rRNA genes showed that strain M46T clustered together with S. canadense CR11T. The genome of strain M46T had a G+C content of 40.6 mol% and chromosome of 6 853 865 bp. Average nucleotide identity (ANI) between strain M46T and S. detergens 6.2 ST and S. siyangense SY1T was 85.1 and 78.1 %, respectively. DNA–DNA relatedness values among strain M46T and other closely related Sphingobacterium species were

Anemoside B4 (AB4) is reported to prevent acute colitis when given via intraperitoneal injection by two recent studies. However, whether oral AB4 protects against chronic colitis which resembles the clinical phenotype of ulcerative colitis (UC) and its mechanism of action are largely unknown.To systemically investigate the effects of oral AB4 against chronic colitis and illustrate the underlying mechanism of action.The preventive, therapeutic, and dose-dependent effects of AB4 against UC were examined in mice with acute or chronic relapsing colitis induced by dextran sulfate sodium (DSS). The inflammatory responses, colonic transcriptome, and 16S rDNA sequencing of the intestinal content of mice were analyzed.Oral administration of AB4 alleviated disease severity and colon shortening in mice with chronic relapsing colitis in a dose-dependent manner. The effects of AB4 were comparable to those of two positive-control compounds: tofacitinib and berberine. Unlike tofacitinib, AB4 did not have a deleterious effect on DSS-induced splenic swelling and anemia. Furthermore, AB4 inhibited the inflammatory responses of colitis, as evidenced by in-vivo, ex-vivo, and in-vitro studies. Transcriptomics revealed that AB4 treatment reversed the DSS-mediated decrease in the expression of colonic Pelo, B3gat2 and Mir8010. In addition, AB4 reversed DSS-induced alterations in the intestinal microbiome in mice. Through fecal microbiota transplantation, we proved that AB4 partially exerted its anti-colitis effects by modulating the gut microbiota.We demonstrated for the first time that AB4 has dose-dependent therapeutic effects against chronic relapsing colitis by modulating the inflammatory response, colonic gene expression, and intestinal microbiota.

In order to better understand the function of a putative GH8 xylanase gene in the xylan-degrading bacterium Paenibacillus physcomitrellae XB, a novel reducing-end xylose-releasing exo-oligoxylanase (Rex) PphRex8A of GH8 family was characterized. Phylogenetic analysis showed that it was clustered tightly with other published GH8 Rexs and exhibited the highest amino acid sequence identity (77.4 %) with the Rex of PbRex8 from P. barengoltzii G22. The three-dimensional (3D) structure of PphRex8A was also built based on the template of PbRex8 (5YXT) and three conserved catalytic active sites (Glu71, Asp263, and Asp129) were predicted and further confirmed by the enzymatic inactivity of their mutants (E71A, D129A, and D263A). The hydrolysis assay of PphRex8A showed that it could hydrolyze xylo-oligosaccharides (XOSs) with a degree of polymerization (DP) ≥ 3, such as xylotriose (X3) through xylohexaose (X6), and some natural XOSs, such as corncob xylan (CCX) and oat spelt xylan (OSX) to release xylose. However, it could not hydrolyze p-nitrophenyl-β-D-xylopyranoside (pNPX), which suggested that it mainly released xylose from the reducing-end of XOSs and belonged to a Rex. In addition, PphRex8A also could deconstruct xylans with high DP, such as wheat arabinoxylan (AX) and beech wood xylan (BWX) to produce XOSs with DP3-6. Moreover, PphRex8A had synergistic effects with other xylanolytic enzymes of P. physcomitrellae XB, such as with PphXyn10 or PphXyn11 at a ratio of 1:3, or with PphXyl43B as a ratio of 3:1, significantly increasing the amounts of reducing sugars toward different xylan substrates. Thus, PphRex8A could be an exo-xylanase toward XOSs and could improve the deconstruction capability of high DP xylans, thereby complementing other xylanolytic enzymes to contribute to xylan degradation and improve the efficiency of converting hemicellulose biomass into energy by P. physcomitrellae XB.

Anemoside B4 (AB4) is a representative component of Pulsatilla decoction that is used in traditional Chinese medicine for treating inflammatory conditions. It is not known whether AB4 has beneficial effects on multiple sclerosis (MS).In the present study, we examined the preventative and therapeutic effects of AB4, and the possible mechanism by which it protects female mice against experimental autoimmune encephalomyelitis (EAE).Preventative treatment with AB4 (given orally at 100 and 200 mg/kg for 18 days) reduced the clinical severity of EAE significantly (from 3.6 ± 1.3 to 1.8 ± 1.5 and 1.6 ± 0.6, respectively), and inhibited demyelination and inflammatory infiltration of the spinal cord. In the therapeutic protocol, oral administration of 200 mg/kg AB4 for 21 days after initiation of EAE significantly alleviated disease severity (from 2.6 ± 1.3 to 0.9 ± 0.6) and was as effective as the clinically used drug fingolimod (0.3 ± 0.6). Furthermore, both doses of AB4 significantly inhibited mRNA expression of TNF-α, IL-6, and IL-17, and STAT3 activation, in the spinal cord; and the ex vivo and iv vitro AB4 treatment markedly inhibited secretion of the three cytokines from lymphocytes of EAE mice upon in vitro restimulation. In addition, AB4 reversed the changes in the composition of the intestinal microbiome observed in EAE mice.We reveal for the first time that AB4 protects against EAE by modulating inflammatory responses and the gut microbiota, demonstrating that AB4 may have potential as a therapeutic agent for treating MS in humans.

BACKGROUND Limited data are available on the discriminatory capacity of quick sequential [sepsis-related] organ failure assessment (qSOFA) versus IDSA/ATS minor criteria for predicting mortality in patients with community-acquired pneumonia (CAP). METHODS An observational prospective cohort study of 2116 patients with CAP was performed. Construct validity was determined using Cronbach α. Discrimination was assessed using the area under the receiver operating characteristic curve (AUROC) and net reclassification improvement (NRI). RESULTS Overall in-hospital mortality was 6.43%. Mortality was 25.96% for patients with a qSOFA score of 2 or higher versus 3.05% for those with a qSOFA score less than 2 (odds ratio for mortality 6.57, P

Natural killer (NK) cells preferentially accumulate at maternal–foetal interface and are believed to play vital immune‐modulatory roles during early pregnancy and related immunological dysfunction may result in pregnant failure such as recurrent miscarriage (RM). However, the mechanisms underlying the establishment of maternal–foetal immunotolerance are complex but clarifying the roles of decidual NK (dNK) cells offers the potential to design immunotherapeutic strategies to assist RM patients. In this report, we analysed RNA sequencing on peripheral NK (pNK) and decidual NK cells during early pregnancy; we identified an immunomodulatory dNK subset CXCR4+CD56brightdNK and investigated its origin and phenotypic and functional characteristics. CXCR4+CD56brightdNK displayed a less activated and cytotoxic phenotype but an enhanced immunomodulatory potential relative to the CXCR4 negative subset. CXCR4+CD56brightdNK promote Th2 shift in an IL‐4‐dependent manner and can be recruited from peripheral blood and reprogramed by trophoblasts, as an active participant in the establishment of immune‐tolerance during early pregnancy. Diminished CXCR4+ dNK cells and their impaired ability to induce Th2 differentiation were found in RM patients and mouse models of spontaneous abortion. Moreover, adoptive transfer of CXCR4+ dNK cells to NK‐deficient (Nfil3–/–) mice showed great therapeutic potential of CXCR4+ dNK via recovering the Th2/Th1 bias and reducing embryo resorption rates. The identification of this new dNK cell subset may lay the foundation for understanding NK cell mechanisms in early pregnancy and provide potential prognostic factors for the diagnosis and therapy of RM., Decidual CXCR4+ NK cell subset is recruited from peripheral blood and reprogramed by trophoblasts. CXCR4+ dNK cells featured with low activity and cytotoxicity but high capacity of inducing Th2 differentiation, are benefit to pregnancy immunotolerance. CXCR4+ dNK cell subset decreases in miscarriage and is less potent to mediate immune tolerance. Adoptive transfer of CXCR4+ dNK cells alleviates pregnancy loss.

The Schistosoma japonicum fatty acid-binding protein (FABP) is used in the cell membrane to absorb and transport fatty acids, which cannot be resynthesized by the organism and combined with hydrophobic ligands. Among the 5 stages of the worm life cycle examined, FABP messenger ribonucleic acid (mRNA) expression was highest in male adult worms, followed by the liver-stage schistosome, and was the lowest in the lung-stage schistosome. The fabp gene-coding region was cloned and expressed to obtain recombinant S. japonicum FABP (rSjFABP) with a molecular weight of approximately 18 kDa. Mice were then immunized against rSjFABP to prepare anti-FABP serum. Using immunohistochemical techniques, FABP protein was found to localize to the eggshell, parenchyma, and digestive tract. Double-stranded RNA-mediated knockdown of FABP mRNA by RNA interference decreased the number of transcripts by >70%. Moreover, the egg production rate decreased, whereas the abnormal egg ratio was significantly increased in the FABP-silenced group compared with the negative control group (P < 0.05). These results demonstrate that FABP localizes in adults and in various stages. FABP contributes to the egg-laying capacity of adults, which may be related to the reproductive function of S. japonicum.

Obesity is prevalent among women of reproductive age and is associated with increased risk of developing multiple pregnancy disorders. Pregnancy must induce immune tolerance to avoid fetal rejection, while obesity can cause chronic inflammation through activating the immune system. Impaired maternal immuno-tolerance leads to pregnancy failure, such as recurrent spontaneous abortion (RSA), one of the most common complications during early pregnancy. How does maternal immune response change under obesity stress in normal pregnancy and RSA? In turn, is obesity affected by different gestational statuses? Limited information is presently available now. Our study investigated pregnancy outcomes and maternal immune responses in two murine models (normal pregnancy and spontaneous abortion models) after obesity challenge with a high-fat diet (HFD). Abortion-prone mice fed HFD had significantly higher weight gains during pregnancy than normal pregnant mice with HFD feeding. Nonetheless, the embryo implantation and resorption rates were comparable between HFD and normal chow diet (NCD)-fed mice in each model. Evaluation of immune cell subsets showed HFD-induced obesity drove the upregulation of activated NK cell-activating receptor (NKp46)+ NK cells and pro-inflammatory macrophages (MHCIIhigh Mφ) as well as CD4+ and CD8+ T cells in the normal pregnancy group. However, in the abortion-prone group, relative more immature NK cells with decreased activity phenotypes were found in obese mice. Moreover, there were increased DCreg (CD11bhigh DC) cells and decreased CD4+ and CD8+ T cells detected in the HFD abortion-prone mice relative to those fed the NCD diet. Our findings reveal how pregnancy obesity and maternal immune regulation are mutually influenced. It is worth noting that the abortion-prone model where active maternal immune status was intensified by obesity, in turn stimulated an overcompensation response, leading to an over-tolerized immune status, and predisposing to potential risks of perinatal complications.

Main differences of incubators are humidity, temperature and gas control ways, which play important roles in regulating the steady state of culture media. In this study, we compared the effects of different types of incubators (air jacket incubators and water jacket incubators) on embryo development and clinical outcomes in human assisted reproduction. We found that temperature recovery time in air jacket incubators was significantly shorter than that in water jacket incubators. Although the O2 recovering time was also significantly shorter in air jacket incubators as compared with the water jacket incubator, no significant differences were observed in CO2 recovering time between two groups, which was also verified by pH recovering time of culture media. Besides, the temperature of culture medium in the dish covered with oil recovered more quickly in the air jacket incubators. However, there were no significant differences observed in the fertilization rate, Day 3 high-quality embryo rate, blastocyst rate, good blastocyst rate and clinical outcomes between two groups. These results indicate that the microenvironment, especially the temperature, in air jacket incubator recover faster than that in water jacket incubators, however, there were no significant differences in embryo development and clinical outcomes between two types of incubators.

To observe the effect ofA total of 120 patients with CRD were randomly divided into an observation group and a control group, 60 cases in each group. Based on the routine treatment of oncology, the patients in the control group were treated with sertraline hydrochloride tablets, 50 mg per time, once a day, and the patients in the observation group were additionally treated withCompared before treatment, the SDS scores and HAMD scores in the two groups were reduced after 2-week, 4-week and 6-week treatment (

Maize-peanut intercropping is an important element of China’s agricultural planting model, as it confers ecological benefits, promotes species diversity, and increases economic efficiency and yield. The aim of this study was to explore the yield differences between intercropping and monoculture, and to determine the mechanism underlying the high yield efficiency of the intercropping system using the 13C isotope tracer labelling method. The early maturing corn hybrid Denghai 618 and the early maturing and high-yielding peanut variety Huayu 22 were used as test materials. Three kinds of planting methods were employed, i.e. the sole maize (SM), the sole peanut (SP) and maize–peanut intercropping (intercropped maize, IM; intercropped peanut, IP), for two consecutive years. IM increased yield by 59.7% and 62.3% comparing with SM in 2015 and 2016, respectively. IP reduced yield by 31.3% and 32.3% comparing with SP in 2015 and 2016, respectively. IM significantly increased the photosynthetic rate, leaf area, 13C assimilation distribution, and dry matter accumulation of summer maize, which led to an increase in kernel number, resulting in an increased yield. The decrease in intercropped peanut yield was mainly caused by a decrease in the percent of plump pod and number of pods per plant. The decrease in peanut yield did not affect the production of intercropping, because of the large intercropping advantage and land equivalence ratio. Maize-peanut intercropping provided greater economic benefits than monoculture. These results showed the utility of the peanut-maize intercropping model.

The success of pregnancy relies on the fine adjustment of the maternal immune system to tolerate the allogeneic fetus. Trophoblasts carrying paternal antigens are the only fetal-derived cells that come into direct contact with the maternal immune cells at the maternal–fetal interface. The crosstalk between trophoblasts and decidual immune cells (DICs) via cell–cell direct interaction and soluble factors such as chemokines and cytokines is a core event contributing to the unique immunotolerant microenvironment. Abnormal trophoblasts–DICs crosstalk can lead to dysregulated immune situations, which is well known to be a potential cause of a series of pregnancy complications including recurrent spontaneous abortion (RSA), which is the most common one. Immunotherapy has been applied to RSA. However, its development has been far less rapid or mature than that of cancer immunotherapy. Elucidating the mechanism of maternal–fetal immune tolerance, the theoretical basis for RSA immunotherapy, not only helps to understand the establishment and maintenance of normal pregnancy but also provides new therapeutic strategies and promotes the progress of immunotherapy against pregnancy-related diseases caused by disrupted immunotolerance. In this review, we focus on recent progress in the maternal–fetal immune tolerance mediated by trophoblasts–DICs crosstalk and clinical application of immunotherapy in RSA. Advancement in this area will further accelerate the basic research and clinical transformation of reproductive immunity and tumor immunity.

The objective is to understand the diversity of bacteria-degrading pesticide pollutants in Xinjiang saline-alkali soil environment and resolve the lack of suitable degrading bacteria resources for bioremediation of pesticide pollution in this environment. The soil of long-term continuous cropping cotton fields in Xinjiang was used to culture the degrading bacterial communities under long-term stress of five pesticides, such as beta-cypermethrin. Then, the degradation rate and structural composition of each bacterial communities were analyzed. The soil bacterial diversity in Xinjiang saline-alkali cotton fields was high, from which not only imidacloprid and other commonly and once used pesticide-degrading bacterial communities were enriched but also isoprocarb-degrading bacterial communities, which had never been used, were enriched. After long-term passage, the structural composition of each degrading bacterial communities was stable, and the degradation rates were between 17 and 48%, respectively, in a specific culture period. Each degrading bacterial communities covers many reported pesticide-degrading bacterial genera and contains unique bacterial genera in each 3. These results laid a foundation for studying the metabolic pathway of pesticide pollutants in saline-alkali environment and exploring microbial resources in Xinjiang. Graphical Abstract Variety of pesticide degrading bacteria resources in saline alkali soil of Xinjiang.

Immunofluorescence staining is a crucial tool for studying the structure and behavior of intracellular proteins and organelles. During the staining process, the permeabilization treatment is usually required to enhance the penetration of a fluorescent antibody into the cells. However, since most of the membrane imaging dyes as well as the membrane lipids will detach from the cell surface after permeabilization, membrane labeling using these dyes is not compatible with immunofluorescence staining. Herein, by linking cholesterol-polyethylene glycol (PEG-Chol) and fluorescein isothiocyanate (FITC) with the amine-rich glycol chitosan (GC), we prepared a multifunctional polymeric construct, GC-PEG Chol-FITC, and realized permeabilization-tolerant plasma membrane imaging. Owing to the presence of abundant amine groups in the labeling reagent and the membrane proteins/lipids, the addition of paraformaldehyde in the fixation step induces the amine-cross-linking between the labeling reagents and the membrane proteins/lipids, thus preventing the detachment of fluorophores from the cell surface after permeabilization. Besides, the large molecular weight effect of the imaging reagent may also account for its antipermeabilization property. Furthermore, by combining immunofluorescence staining with the plasma membrane labeling by GC-PEG Chol-FITC, we simultaneously imaged the plasma membrane and cytoskeletons, and clearly observed metaphase cells and binucleated cells. The concept of using amine-rich polymeric dyes for plasma membrane imaging will inspire the development of more permeabilization-resistant membrane labeling dyes with better performance, which can realize simultaneous membrane and intracellular protein imaging and facilitate the future studies of membrane-intracellular protein interactions.

BackgroundThe strain Paenibacillus physcomitrellae XB isolated from moss of Physcomitrella patens was found have the xylan degradation ability, but its degradation characteristics and the related mechanism has not been revealed.ResultsIn this study, Paenibacillus physcomitrellae XB exhibited different xylan degradation ability under the different substrates of corncob xylan (CCX), oat spet xylan (OSX), wheat flour arabinoxylan (AX) and beech wood xylan (BWX). Genomic analysis showed that ~ 38 genes were related to xylan degradation, and quantitative real time RT-PCR showed that two glycoside hydrolase family 43 genes (Pph_0602 and Pph_2344) were up-regulated on 1% CCX and xylose. Substrate-specific experiments with purified proteins Ppxyl43A (Pph_0602) and Ppxyl43B (Pph_2344) revealed that both of them exhibited β-xylosidase activity toward chromogenic substrate p-nitrophenyl–D-xylopyranoside and α-L-arabinofuranosidase activity toward p-nitrophenyl-α-L-arabinofuranoside, indicating at least bifunctionality. Combined their degradation features on the natural substrates of different xylans with the hydrolytic products separated by thin-layer chromatography and high-performance anion exchange chromatography profiles, it was found that both Ppxyl43A and Ppxyl43B were with the similar degradation ability on xylo-oligosaccharides (like CCX, OSX, xylohexaose and xylobiose). Both of them even could hydrolyze xylohexaose and xylobiose completely to xylose, but could not hydrolyze BWX and AX to produce xylooligosaccharides or xylose, suggesting they have no endo-xylanase activity and mainly hydrolyze xylo-oligosaccharides by β-xylosidase activity. Moreover, the kinetic parameters of β-xylosidase and α-L-arabinofuranosidase of both two proteins indicated their affinity with all the detected natural substrate (CCX) and chromogenic substrates were nearly similar. In addition, despite having no signal peptides, both of them might export outside the cell by the nonconventional pathways. However, Ppxy143B exhibited wider temperature and pH ranges, higher pH and thermostability, and was less influenced by metal ions than Ppxyl43A. Given its enzymatic characteristics and predicted structure, it is likely that the C-terminus domain (GH43_C2) of Ppxyl43B enhances the stability of the two enzymes and also restricts the substrates’ or metal ions’ access to the active sites.ConclusionsPpxyl43A and Ppxyl43B were β-xylosidase/α-L-arabinofuranosidase bifunctional enzymes with different structures from Paenibacillus physcomitrellae XB and exhibited similar xylo-oligosaccharides hydrolyse ability, which would be useful in the further lignocellulosic biomass conversion.