Discovery of novel aporphine alkaloid derivative as potent TLR2 antagonist reversing macrophage polarization and neutrophil infiltration against acute inflammation.

Toll-like receptor 2 (TLR2) mediated macrophages regulate the protective immune response to infectious microorganisms, but the aberrant activation of macrophages often leads to pathological inflammation, including tissue damage. In this study, we identified antagonists of TLR2 by screening 2100 natural products and subsequently identified Taspine, an aporphine alkaloid, as an excellent candidate. Furthermore, analysis of the 10 steps chemical synthesis route and structural optimization yielded the Taspine derivative SMU-Y6, which has higher activity, better solubility, and improved drug-feasible property. Mechanistic studies and seq-RNA analysis revealed that SMU-Y6 inhibited TLR2 over other TLRs, hindered the formation of TLR2/MyD88 complex, and blocked the downstream NF-κB and MAPK signaling pathway, thus suppressing the release of inflammatory cytokines. SMU-Y6 could stabilize TLR2 and bind to TLR2 protein with a Kd of 0.18 µmol/L. Additionally, SMU-Y6 could efficiently reverse the M1 phenotype macrophage polarization, reduce the production of cytokines as well as infiltration of neutrophiles and alleviate the local inflammation in mice with acute paw edema and colitis. Collectively, we reported the first aporphine alkaloid derivative that selectively inhibits TLR2 with high binding affinity and superior drug-feasible property, thus providing an urgently-needed molecular probe and potential drug candidate for inflammatory and autoimmune disease therapy.

Five Rice Seed-Specific NF-YC Genes Redundantly Regulate Grain Quality and Seed Germination via Interfering Gibberellin Pathway.

NF-YCs are important transcription factors with diverse functions in the plant kingdoms including seed development. NF-YC8, 9, 10, 11 and 12 are close homologs with similar seed-specific expression patterns. Despite the fact that some of the NF-YCs are functionally known; their biological roles have not been systematically explored yet, given the potential functional redundancy. In this study, we generated pentuple mutant pnfyc of NF-YC8-12 and revealed their functions in the regulation of grain quality and seed germination. pnfyc grains displayed significantly more chalkiness with abnormal starch granule packaging. pnfyc seed germination and post-germination growth are much slower than the wild-type NIP, largely owing to the GA-deficiency as exogenous GA was able to fully recover the germination phenotype. The RNA-seq experiment identified a total of 469 differentially expressed genes, and several GA-, ABA- and grain quality control-related genes might be transcriptionally regulated by the five NF-YCs, as revealed by qRT-PCR analysis. The results demonstrated the redundant functions of NF-YC8-12 in regulating GA pathways that underpin rice grain quality and seed germination, and shed a novel light on the functions of the seed-specific NF-YCs.

Hypoxia-inducible factor-1α activation can attenuate renal podocyte injury and alleviate proteinuria in rats in a simulated high-altitude environment.

OBJECTIVE: The aims of this study were to understand whether podocyte injury is involved in proteinuria after rapid ascent to high altitude and to explore whether hypoxia-inducible factor (HIF)-1α is involved in the adaptive regulation of this proteinuria. METHODS: Rats in the experimental group were housed in a low-pressure oxygen chamber to simulate a high-altitude environment (5,000 m). The intervention group was placed under the same conditions as the experimental group and prolyl-hydroxylase inhibitor (PHI) was intraperitoneally injected. The control group was housed in a low altitude environment (500 m). On days 0, 7, 14, and 28, urinary albumin quantification and electrophoresis were performed. The expression levels of CD2-associated protein (CD2AP), nephrin and HIF-1α were detected by immunofluorescence. RESULTS: The medium and large molecule proteins with molecular weights ranging from 63 to 75 kD were present in the urine of rats in the experimental group and that the urinary albumin levels first increased and then decreased with time and the increase on day 14 was most significant (24.58 ± 4.30 mg on day 14 VS 5.13 ± 1.58 mg on day 0). Electron microscopy revealed podocyte lesions in rats in the experimental group. Immunofluorescence results showed that the protein expression levels of CD2AP and nephrin in the glomeruli of rats in the experimental group were lower than those in the control group (P < 0.001) and that the expression levels of which in the intervention group were higher than those in the experimental group (P < 0.001). The expression of HIF-1α protein in the renal tissues of rats in the experimental group was higher than that in the control group (P < 0.001) and lower than that in the intervention group (P < 0.001). CONCLUSION: The podocyte injury may be involved in the occurrence of proteinuria after rapid ascent to high altitude. PHI may have a potential role in reducing proteinuria by upregulating local HIF-1α expression in the kidney to alleviate podocyte injury.

Comparative transcriptome analysis of the cold resistance of the sterile rice line 33S.

Rice (Oryza sativa L.) is one of the most important species for food production worldwide. Low temperature is a major abiotic factor that affects rice germination and reproduction. Here, the underlying regulatory mechanism in seedlings of a TGMS variety (33S) and a cold-sensitive variety (Nipponbare) was investigated by comparative transcriptome. There were 795 differentially expressed genes (DEGs) identified only in cold-treated 33S, suggesting that 33S had a unique cold-resistance system. Functional and enrichment analysis of these DEGs revealed that, in 33S, several metabolic pathways, such as photosynthesis, amino acid metabolism, secondary metabolite biosynthesis, were significantly repressed. Moreover, pathways related to growth and development, including starch and sucrose metabolism, and DNA biosynthesis and damage response/repair, were significantly enhanced. The expression of genes related to nutrient reserve activity were significantly up-regulated in 33S. Finally, three NAC and several ERF transcription factors were predicted to be important in this transcriptional reprogramming. This present work provides valuable information for future investigations of low-temperature response mechanisms and genetic improvement of cold-tolerant rice seedlings.

Effectiveness and safety of endoscopy-assisted versus conventional open lateral neck dissection: A meta-analysis.

The objective of this study was to systematically evaluate the effectiveness and safety of endoscopic-assisted lateral neck dissection (EALND) compared with conventional open lateral neck dissection (COLND) for the treatment of thyroid cancer with positive lymph node metastases. Medical literature databases including PubMed, Embase, the Cochrane Library, CNKI, Wan Fang and VIP were systematically searched for articles that compared EALND and COLND for the treatment of thyroid carcinoma with lymph node metastasis, up to June 2019. The quality of included studies was evaluated using the Newcastle-Ottawa Scale (NOS). Meta-analysis was performed using RevMan 5.3 software after two evaluators independently screened the literature, extracted information and evaluated the methodological quality of included studies according to inclusion and exclusion criteria, resulting in the selection of seven studies with a total of 372 patients from six non-RCTs and an RCT. The results of meta-analysis showed that EALND was associated with a longer operative time (MD = 24.86, 95∗CI：21.76 to 27.96, P＜0.05), with a shorter postoperative stay (MD = -1.45, 95％CI：-2.70 to -0.21，P = 0.02), reduced length of scar (MD = -8.14，95％CI：-8.41 to -7.88, P＜0.00001) and a lower incidence of neck discomfort (OR = 0.19, 95％CI：0.07 to 0.58, P = 0.003) compared with COLND. The incidences in both groups of transient hypocalcemia (OR = 0.66，95％CI：0.28 to 1.55，P = 0.343), transient hoarseness (OR = 0.58，95％CI：0.17 to 1.93，P = 0.38)，chylous fistula (OR = 0.69，95％CI：0.26 to 1.83，P = 0.45), choking on water (OR = 0.24，95％CI：0.04 to 1.31，P = 0.10) and the number of lymph nodes retrieved from the lateral cervical region (MD = 0.14，95％CI：-0.36 to 0.65，P = 0.59) were not statistically significant. It was concluded that EALND was safe and feasible compared with COLND, despite the longer operation time. The incision was more aesthetically pleasing and the postoperative recovery was quicker, which makes EALND a clinical procedure worthy of use in such cases.

Harvesting the vibration energy of α-MnO2 nanostructures for complete catalytic oxidation of carcinogenic airborne formaldehyde at ambient temperature.

Vibration is one of the most prevalent energy sources in natural environment, which can also be harvested and utilized to drive chemical reaction. Herein, mechanical vibration is used for enhancing the catalytic decomposition of formaldehyde at ambient temperature with the assistance of four well-defined morphologies α-MnO2 (nanowire, nanotube, nanorod and nanoflower). In particular, α-MnO2 nanowire exhibits the best catalytic activity, which can completely mineralize formaldehyde into carbon dioxide at ambient temperature by harvesting the vibration energy. To the best of our knowledge, this may be the first report that α-MnO2, as a non-noble metal catalyst, can completely decompose formaldehyde to carbon dioxide at ambient temperature. The characterization results show that α-MnO2 nanowire has a much higher oxygen vacancy concentration than other three catalysts. In addition, thermal effect generated from friction between nanoparticles induced by ultrasonic vibration may enhance its catalytic activity. More importantly, it is the vibration that effectively promotes the activation of O2 adsorbed on the surface oxygen vacancy to produce more , thus increasing the catalytic decomposition performance. The strategy presented herein demonstrates a new approach for efficient use of mechanical vibration to improve catalytic activity of traditional catalysts.

A challenging therapeutic method for breast cancer: Non-lipolytic endoscopic axillary surgery through periareolar incisions.

Surgical treatment of breast cancer is becoming increasingly precise, less invasive, and more cosmetically pleasing. Sentinel lymph node biopsy (SLNB) and axillary lymph node dissection (ALND) remain the standard treatment methods for breast cancer. However, these methods still require incisions in the breasts or axilla. Mastoscopic axillary lymph node dissection (MALND) surgery, although first reported several years ago, has not been widely used as it involves lipolysis. Non-lipolytic mastoscopy may be more appealing; however, the lack of a cavity in the breast and the abundant fat and glands make this procedure challenging. In addition, incision of the trocar in the axilla has been shown to have no advantage over traditional breast-conserving surgery. The present study describes 16 cases of non-lipolytic endoscopic axillary surgery without incisions in the axilla.

WGCNA Analysis of Salt-Responsive Core Transcriptome Identifies Novel Hub Genes in Rice.

Rice, being a major staple food crop and sensitive to salinity conditions, bears heavy yield losses due to saline soil. Although some salt responsive genes have been identified in rice, their applications in developing salt tolerant cultivars have resulted in limited achievements. Herein, we used bioinformatic approaches to perform a meta-analysis of three transcriptome datasets from salinity and control conditions in order to reveal novel genes and the molecular pathways underlying rice response to salt. From a total of 28,432 expressed genes, we identify 457 core differentially expressed genes (DEGs) constitutively responding to salt, regardless of the stress duration, genotype, or the tissue. Gene co-expression analysis divided the core DEGs into three different modules, each of them contributing to salt response in a unique metabolic pathway. Gene ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses highlighted key biological processes and metabolic pathways involved in the salt response. We identified important novel hub genes encoding proteins of different families including CAM, DUF630/632, DUF581, CHL27, PP2-13, LEA4-5, and transcription factors, which could be functionally characterized using reverse genetic experiments. This novel repertoire of candidate genes related to salt response in rice will be useful for engineering salt tolerant varieties.

Highly Stable Mesoporous Luminescence-Functionalized MOF with Excellent Electrochemiluminescence Property for Ultrasensitive Immunosensor Construction.

In this work, a novel mesoporous luminescence-functionalized metal-organic framework (Ru-PCN-777) with high stability and excellent electrochemiluminescence (ECL) performance was synthesized by immobilizing Ru(bpy)2(mcpbpy)2+ on the Zr6 cluster of PCN-777 via a strong coordination bond between Zr4+ and -COO-. Consequently, the Ru(bpy)2(mcpbpy)2+ could not only cover the surface of PCN-777 but also graft into the interior of PCN-777, which greatly increased the loading amount of Ru(bpy)2(mcpbpy)2+ and effectively prevented the leaching of the Ru(bpy)2(mcpbpy)2+ resulting in a stable and high ECL response. Considering the above merits, we utilized the mesoporous Ru-PCN-777 to construct an ECL immunosensor to detect mucin 1 (MUC1) based on proximity-induced intramolecular DNA strand displacement (PiDSD). The ECL signal was further enhanced by the enzyme-assisted DNA recycling amplification strategy. As expected, the immunosensor had excellent sensitivity, specificity, and responded wide linearly to the concentration of MUC1 from 100 fg/mL to 100 ng/mL with a low detection limit of 33.3 fg/mL (S/N = 3). It is the first time that mesoporous Zr-MOF was introduced into ECL system to assay biomolecules, which might expand the application of mesoporous metal-organic frameworks (MOFs) in bioanalysis. This work indicates that the use of highly stable mesoporous luminescence-functionalized MOFs to enhance the ECL intensity and stability is a feasible strategy for designing and constructing high-performance ECL materials, and therefore may shed light on new ways to develop highly sensitive and selective ECL sensors.

Long non-coding RNA ANRIL promotes carcinogenesis via sponging miR-199a in triple-negative breast cancer.

Triple-negative breast cancer (TNBC) is a complex breast cancer subtype characterized by the absence of estrogen receptor (ER), progesterone receptor (PR) and human epidermal growth factor receptor 2 (Her2). Long non-coding RNA (lncRNA) antisense non-coding RNA in the INK4 locus (ANRIL) has been verified as oncogenic molecular in series of tumors, however, the role of ANRIL in TNBC carcinogenesis is still unclear. The purpose of present study is to investigate the expression and in-depth regulation of ANRIL on TNBC tumorigenesis. Expression level of ANRIL was up-regulated in TNBC tumor tissue and cell lines compared to noncancerous tissue and non-TNBC cells. Besides, the up-regulated ANRIL expression was closely correlated to poor prognosis. In vitro, loss-of-function experiments showed that ANRIL knockdown interfered by interference oligonucleotide could markedly suppress TNBC cells proliferation and enhance apoptosis. In vivo, ANRIL knockdown inhibited the tumor growth. Bioinformatics analysis and luciferase reporter assay revealed that miR-199a targeted ANRIL at 3'-UTR. Rescue experiments showed that miR-199a inhibitor could reverse the tumor-suppressing role of ANRIL knockdown on TNBC proliferation and apoptosis. Overall, present study demonstrated that ANRIL overexpression modulated TNBC tumorigenesis through acting as molecular 'sponge' for miR-199a, providing a novel insight and therapeutic target for TNBC.

The RhoGAP SPIN6 associates with SPL11 and OsRac1 and negatively regulates programmed cell death and innate immunity in rice.

The ubiquitin proteasome system in plants plays important roles in plant-microbe interactions and in immune responses to pathogens. We previously demonstrated that the rice U-box E3 ligase SPL11 and its Arabidopsis ortholog PUB13 negatively regulate programmed cell death (PCD) and defense response. However, the components involved in the SPL11/PUB13-mediated PCD and immune signaling pathway remain unknown. In this study, we report that SPL11-interacting Protein 6 (SPIN6) is a Rho GTPase-activating protein (RhoGAP) that interacts with SPL11 in vitro and in vivo. SPL11 ubiquitinates SPIN6 in vitro and degrades SPIN6 in vivo via the 26S proteasome-dependent pathway. Both RNAi silencing in transgenic rice and knockout of Spin6 in a T-DNA insertion mutant lead to PCD and increased resistance to the rice blast pathogen Magnaporthe oryzae and the bacterial blight pathogen Xanthomonas oryzae pv. oryzae. The levels of reactive oxygen species and defense-related gene expression are significantly elevated in both the Spin6 RNAi and mutant plants. Strikingly, SPIN6 interacts with the small GTPase OsRac1, catalyze the GTP-bound OsRac1 into the GDP-bound state in vitro and has GAP activity towards OsRac1 in rice cells. Together, our results demonstrate that the RhoGAP SPIN6 acts as a linkage between a U-box E3 ligase-mediated ubiquitination pathway and a small GTPase-associated defensome system for plant immunity.

Intervention of childhood and adolescents obesity in Shantou city.

OBJECTIVES: The purpose of this study was to evaluate the effects of multicomponent school based intervention constituted of diet modification, regular exercise and psychosocial consultation on body status in overweight and obese children and adolescents. And to come up with an appropriate intervention protocol for controlling children and adolescents obesity in Shantou city. METHODS: Two schools were randomised to intervention group and control group respectively. A total of 41 students enrolled were diagnosed as overweight or obese. Twenty-six students of the intervention group completed the one-year intervention programme consisted of diet modification, regular exercise and psychological consultation except two of them were transferred to another school. The differences of BMI, anthropometric measures, metabolic profile and the scores of questionnaire and the scale were compared to evaluate the effects of the intervention programme. RESULTS: After one-year intervention, it observed in the intervention group that BMI-Z score, WHR and WHtR had significant improvements, and there was a nonsignificant trend (P=0.053) for a decrease in BMI-P. Fasting plasma glucose, cholesterol (CH) and low-density-lipoprotein cholesterol (LDL-C) levels in the intervention group showed nonsignificant trend for a decrease (PFPG=0.084, PCH=0.057, PLDLC=0.098), compared with a significant increase of triglycerides (TG) and LDL-C levels in the control group (PTG=0.041, PLDL-C=0.038). There were some positive dietary, physical activity, or sedentary behaviour changes found in the students of the intervention group as the scores of the questionnaire got significant improvement (P=0.04). CONCLUSIONS: Our one-year multicomponent school-based intervention programme did have positive effects to some extents on health state and lifestyle behaviour of overweight and obese children and adolescents, which indicated that it is feasible and important to implement such a school-based intervention programme in Shantou city.

Characterization and fine mapping of the rice blast resistance gene Pia.

Blast, caused by Magnaporthe oryzae, is one of the most widespread and destructive diseases of rice. Breeding durable resistant cultivars (cvs) can be achieved by pyramiding of various resistance (R) genes. Pia, carried by cv. Aichi Asahi, was evaluated against 612 isolates of M. oryzae collected from 10 Chinese provinces. The Pia gene expresses weak resistance in all the provinces except for Jiangsu. Genomic position-ready marker-based linkage analysis was carried out in a mapping population consisting of 800 F(2) plants derived from a cross of Aichi Asahi×Kasalath. The locus was defined in an interval of approximately 90 kb, flanked by markers A16 and A21. Four candidate genes (Pia-1, Pia-2, Pia-3, and Pia-4), all having the R gene conserved structure, were predicted in the interval using the cv. Nipponbare genomic sequence. Four candidate resistance gene (CRG) markers (A17, A25, A26, and A27), derived from the four candidates, were subjected to genotyping with the recombinants detected at the flanking markers. The first three markers completely co-segregated with the Pia locus, and the fourth was absent in the Aichi Asahi genome and disordered with the Pia locus and its flanking markers, indicating that the fourth candidate gene, Pia-4, could be excluded. Co-segregation marker-based genotyping of the three sets of differentials with known R gene genotypes revealed that the genotype of A26 (Pia-3) perfectly matched the R gene genotype of Pia, indicating that Pia-3 is the strongest candidate gene for Pia.

The Pik-p resistance to Magnaporthe oryzae in rice is mediated by a pair of closely linked CC-NBS-LRR genes.

The blast resistance gene Pik-p, mapping to the Pik locus on the long arm of rice chromosome 11, was isolated by map-based in silico cloning. Four NBS-LRR genes are present in the target region of cv. Nipponbare, and a presence/absence analysis in the Pik-p carrier cv. K60 excluded two of these as candidates for Pik-p. The other two candidates (KP3 and KP4) were expressed in cv. K60. A loss-of-function experiment by RNAi showed that both KP3 and KP4 are required for Pik-p function, while a gain-of-function experiment by complementation test revealed that neither KP3 nor KP4 on their own can impart resistance, but that resistance was expressed when both were introduced simultaneously. Both Pikp-1 (KP3) and Pikp-2 (KP4) encode coiled-coil NBS-LRR proteins and share, respectively, 95 and 99% peptide identity with the two alleles, Pikm1-TS and Pikm2-TS. The Pikp-1 and Pikp-2 sequences share only limited homology. Their sequence allowed Pik-p to be distinguished from Pik, Pik-s, Pik-m and Pik-h. Both Pikp-1 and Pikp-2 were constitutively expressed in cv. K60 and only marginally induced by blast infection.

The isolation and characterization of Pik, a rice blast resistance gene which emerged after rice domestication.

• The rice-rice blast pathosystem is of great interest, not only because of the damaging potential of rice blast to the rice crop, but also because both the pathogen and its host are experimentally amenable. The rice blast resistance gene Pik, which is one of the five classical alleles located at the Pik locus on the long arm of chromosome 11, confers high and stable resistance to many Chinese rice blast isolates. • The isolation and functional characterization of Pik were performed in the present study through genetic and genomic approaches. • A combination of Pik-1 and Pik-2 is required for the expression of Pik resistance. Both Pik-1 and Pik-2 encode coiled-coil nucleotide binding site leucine-rich repeat (NBS-LRR) proteins, and each shares a very high level of protein identity with corresponding proteins encoded by the Pik-m and Pik-p alleles. Pik could be distinguished from other Pik alleles, including Pik-m and Pik-p, by the allele-specific, single-nucleotide polymorphism T1-2944G. • The coupled genes probably did not evolve as a result of a duplication event, and are far from any NBS-LRR R gene characterized. Pik is a younger allele at the locus that probably emerged after rice domestication.