Genome-wide analysis of the peanut CaM/CML gene family reveals that the AhCML69 gene is associated with resistance to Ralstonia solanacearum.

BACKGROUND: Calmodulins (CaMs)/CaM-like proteins (CMLs) are crucial Ca2+-binding sensors that can decode and transduce Ca2+ signals during plant development and in response to various stimuli. The CaM/CML gene family has been characterized in many plant species, but this family has not yet been characterized and analyzed in peanut, especially for its functions in response to Ralstonia solanacearum. In this study, we performed a genome-wide analysis to analyze the CaM/CML genes and their functions in resistance to R. solanacearum. RESULTS: Here, 67, 72, and 214 CaM/CML genes were identified from Arachis duranensis, Arachis ipaensis, and Arachis hypogaea, respectively. The genes were divided into nine subgroups (Groups I-IX) with relatively conserved exon‒intron structures and motif compositions. Gene duplication, which included whole-genome duplication, tandem repeats, scattered repeats, and unconnected repeats, produced approximately 81 pairs of homologous genes in the AhCaM/CML gene family. Allopolyploidization was the main reason for the greater number of AhCaM/CML members. The nonsynonymous (Ka) versus synonymous (Ks) substitution rates (less than 1.0) suggested that all homologous pairs underwent intensive purifying selection pressure during evolution. AhCML69 was constitutively expressed in different tissues of peanut plants and was involved in the response to R. solanacearum infection. The AhCML69 protein was localized in the cytoplasm and nucleus. Transient overexpression of AhCML69 in tobacco leaves increased resistance to R. solanacearum infection and induced the expression of defense-related genes, suggesting that AhCML69 is a positive regulator of disease resistance. CONCLUSIONS: This study provides the first comprehensive analysis of the AhCaM/CML gene family and potential genetic resources for the molecular design and breeding of peanut bacterial wilt resistance.

Disease-Specific Autoantibodies Induce Trained Immunity in RA Synovial Tissues and Its Gene Signature Correlates with the Response to Clinical Therapy.

Much evidence suggests that trained immunity is inappropriately activated in the synovial tissue in rheumatoid arthritis (RA), but the underlying mechanism remains unclear. Here, we describe how RA-specific autoantibody deposits can train human monocytes to exert the hyperactive inflammatory response, particularly via the exacerbated release of tumor necrosis factor α (TNFα). Comparative transcriptomic analysis by plate-bound human IgG (cIgG) or ß-glucan indicated that metabolic shift towards glycolysis is a crucial mechanism for trained immunity. Moreover, the cIgG-trained gene signatures were enriched in synovial tissues from patients with ACPA- (anticitrullinated protein antibody-) positive arthralgia and undifferentiated arthritis, and early RA and established RA bore a great resemblance to the myeloid pathotype, suggesting a historical priming event in vivo. Additionally, the expression of the cIgG-trained signatures is higher in the female, older, and ACPA-positive populations, with a predictive role in the clinical response to infliximab. We conclude that RA-specific autoantibodies can train monocytes in the inflamed lesion as early as the asymptomatic stage, which may not merely improve understanding of disease progression but may also suggest therapeutic and/or preventive strategies for autoimmune diseases.

ZBTB24 regulates the apoptosis of human T cells via CDCA7/TRAIL-receptor axis.

Mutations in ZBTB24 and CDCA7 cause the Immunodeficiency, Centromeric Instability and Facial Anomalies syndrome type 2 and 3 (ICF2/3), respectively. Most ICF2 patients carry ZBTB24 nonsense mutations and are thus ZBTB24-deficient. Although the immune deficiency in ICF2 patients is primarily regarded as a B-cell defect due to the greatly reduced serum antibodies and circulating memory B cells, the reduced expansions of PBMCs stimulated by mitogens or recall antigens suggest a T-cell defect in these patients as well. However, the molecular mechanisms behind this T-cell dysfunction remain unknown. In the present study, we demonstrated that ZBTB24-deficiency significantly represses the proliferation of human T cells by promoting TRAIL-induced cell death. Downregulation of ZBTB24 in both Jurkat and human primary T cells upregulates the expression of TRAIL and/or its death receptors (TRAIL-R1/2), and induces significant amount of cells to undergo apoptosis. The profound survival defects of ZBTB24-deficient cells are largely reversed by blocking TRAIL/TRAIL-R interactions with exogenous recombinant TRAIL-R2. Moreover, ZBTB24-downregulation reduces the expression of CDCA7, and knockdown of the latter in human T cells results in a phenotype resembling that caused by ZBTB24-depletion. Functionally, overexpression of CDCA7 abrogates the increased apoptosis in ZBTB24-depleted Jurkat T cells. Together, these data indicated that ZBTB24 regulates human T-cell apoptosis via CDCA7/TRAIL-R axis. Our study thus not only provides a molecular explanation for the T-cell defects in ZBTB24-deficient ICF2 patients, but also highlights a convergence between ZBTB24 and CDCA7, the two ICF genes, in modulating the functions of T cells.

[Establishment of early warning control charts based on the syndromic surveillance data of outpatient diarrhea in Beijing].

OBJECTIVE: To establish the control charts for early warning of diarrhea based on the syndromic surveillance data from enteric clinic in Beijing. METHODS: The outpatient data from enteric clinic of a Grade Three General hospital in Haidian district, Beijing from April 1 to Oct. 31, 2009 and from May 1 to Nov.10, 2010 were collected, according to the moving average method, the baseline calculated, the value of probability α and µα, the early warning value based on the formula "w=Xj+µαSj" calculated and the early warning control charts drew at last. RESULTS: According to the harmfulness, the severity and controllability of diarrheal diseases, the value of probability α was determined as 0.01, then µα (unilateral) as 2, based on the early warning value, the control charts of diarrheal diseases, bacillary dysentery and other infectious diarrhea were established. CONCLUSION: The enteric clinic requires to further collect baseline data to evaluate and continuously adjust the established control charts for the best early warning model in accordance with the enteric clinic.

Prepregnancy body mass index and gestational weight gain with the outcome of pregnancy: a 13-year study of 292,568 cases in China.

PURPOSE: We aimed to investigate the combined associations of prepregnancy body mass index (BMI) and gestational weight gain (GWG) with pregnancy outcomes in Chinese women. METHODS: Data for 292,568 singleton term pregnancies were selected from 1993 to 2005 based on the Perinatal Health Care Surveillance System, with anthropometric measurements being collected prospectively. Prepregnancy BMI was categorized according to the definitions of the World Health Organization (WHO). Total GWG was categorized into four groups. Adjusted associations of prepregnancy BMI and GWG with outcomes of interest were estimated using logistic regression analyses. GWG was categorized as below, within and above the Institute of Medicine (IOM) (2009) recommendations. RESULTS: Maternal overweight and high GWG or GWG above the IOM recommendation were associated with hypertensive disorders complicating pregnancy, cesarean delivery, macrosomia and large-for-gestational-age (LGA) infants. Maternal underweight and low GWG or GWG below the IOM recommendation were risk factors for low-birth-weight (LBW) and small-for-gestational-age (SGA) infants. Moreover, being overweight [odds ratio (OR) 1.2, 95 % confidence interval (CI) 1.0-1.3) and having a low weight gain (OR 1.1, 95 % CI 1.0-1.1) increased the risk of newborn asphyxia. CONCLUSION: Being overweight/obese and having a high weight gain, as well as being underweight and having a low weight gain, were associated with increased risks for adverse pregnancy outcomes in Chinese women.

Complete Genome Sequence Analysis of Ralstonia solanacearum Strain PeaFJ1 Provides Insights Into Its Strong Virulence in Peanut Plants.

The bacterial wilt of peanut (Arachis hypogaea L.) caused by Ralstonia solanacearum is a devastating soil-borne disease that seriously restricted the world peanut production. However, the molecular mechanism of R. solanacearum-peanut interaction remains largely unknown. We found that R. solanacearum HA4-1 and PeaFJ1 isolated from peanut plants showed different pathogenicity by inoculating more than 110 cultivated peanuts. Phylogenetic tree analysis demonstrated that HA4-1 and PeaFJ1 both belonged to phylotype I and sequevar 14M, which indicates a high degree of genomic homology between them. Genomic sequencing and comparative genomic analysis of PeaFJ1 revealed 153 strain-specific genes compared with HA4-1. The PeaFJ1 strain-specific genes consisted of diverse virulence-related genes including LysR-type transcriptional regulators, two-component system-related genes, and genes contributing to motility and adhesion. In addition, the repertoire of the type III effectors of PeaFJ1 was bioinformatically compared with that of HA4-1 to find the candidate effectors responsible for their different virulences. There are 79 effectors in the PeaFJ1 genome, only 4 of which are different effectors compared with HA4-1, including RipS4, RipBB, RipBS, and RS_T3E_Hyp6. Based on the virulence profiles of the two strains against peanuts, we speculated that RipS4 and RipBB are candidate virulence effectors in PeaFJ1 while RipBS and RS_T3E_Hyp6 are avirulence effectors in HA4-1. In general, our research greatly reduced the scope of virulence-related genes and made it easier to find out the candidates that caused the difference in pathogenicity between the two strains. These results will help to reveal the molecular mechanism of peanut-R. solanacearum interaction and develop targeted control strategies in the future.

Neutrophils as regulators of macrophage-induced inflammation in a setting of allogeneic bone marrow transplantation.

Clinical data reveal that patients with allogeneic hematopoietic stem cell transplantation (HSCT) are vulnerable to infection and prone to developing severe sepsis, which greatly compromises the success of transplantation, indicating a dysregulation of inflammatory immune response in this clinical setting. Here, by using a mouse model of haploidentical bone marrow transplantation (haplo-BMT), we found that uncontrolled macrophage inflammation underlies the pathogenesis of both LPS- and E.coli-induced sepsis in recipient animals with graft-versus-host disease (GVHD). Deficient neutrophil maturation in GVHD mice post-haplo-BMT diminished modulation of macrophage-induced inflammation, which was mechanistically dependent on MMP9-mediated activation of TGF-ß1. Accordingly, adoptive transfer of mature neutrophils purified from wild-type donor mice inhibited both sterile and infectious sepsis in GVHD mice post-haplo-BMT. Together, our findings identify a novel mature neutrophil-dependent regulation of macrophage inflammatory response in a haplo-BMT setting and provide useful clues for developing clinical strategies for patients suffering from post-HSCT sepsis.

Membrane Bound CRT Fragment Accelerates Tumor Growth of Melanoma B16 Cell In Vivo through Promoting M2 Polarization via TLR4.

Calreticulin (CRT) is a major calcium-binding luminal resident protein on the endoplasmic reticulum that can also be released extracellular as well as anchored on surface of cells. Previously, we demonstrated that soluble recombinant CRT fragment 39-272 (CRT/39-272) exhibited potent immunostimulatory effects as well as immunoregulation effects on immune cells. Here, we constructed stable B16 melanoma cell lines expressing recombinant CRT/39-272 on the membrane (B16-tmCRT/39-272) to investigate the roles of cell surface CRT on tumor progression. We found that B16-tmCRT/39-272 cells subcutaneously inoculated into C57BL/6 mice exhibited stronger tumorigenicity than the B16-EGFP control cells. The tumor associated macrophages infiltrated in tumors were mainly M2 phenotype. Regulatory T cells (Tregs) were also expanded more in bearing mice. Consistent with the in vivo results, B16-tmCRT/39-272 promoted macrophage polarization toward F4/80+CD206+ M2 macrophages and promoted transforming growth factor beta (TGF-ß) secretion in vitro, which could promote naïve CD4+ T cell differentiation into Tregs. These results imply that the tmCRT/39-272 could accelerate tumor development by enhancing M2 macrophage polarization to induce TGF-ß secretion, and then promoted Treg differentiation in the tumor microenvironment. Our data may provide useful clues for better understanding of the potentiating roles of CRT in tumorigenesis.

Comparative transcriptome analysis revealed molecular mechanisms of peanut leaves responding to Ralstonia solanacearum and its type III secretion system mutant.

Bacterial wilt caused by Ralstonia solanacearum is a serious soil-borne disease that limits peanut production and quality, but the molecular mechanisms of the peanut response to R. solanacearum remain unclear. In this study, we reported the first work analyzing the transcriptomic changes of the resistant and susceptible peanut leaves infected with R. solanacearum HA4-1 and its type III secretion system mutant strains by the cutting leaf method at different timepoints (0, 24, 36, and 72 h post inoculation). A total of 125,978 differentially expressed genes (DEGs) were identified and subsequently classified into six groups to analyze, including resistance-response genes, susceptibility-response genes, PAMPs induced resistance-response genes, PAMPs induced susceptibility-response genes, T3Es induced resistance-response genes, and T3Es induced susceptibility-response genes. KEGG enrichment analyses of these DEGs showed that plant-pathogen interaction, plant hormone signal transduction, and MAPK signaling pathway were the outstanding pathways. Further analysis revealed that CMLs/CDPKs-WRKY module, MEKK1-MKK2-MPK3 cascade, and auxin signaling played important roles in the peanut response to R. solanacearum. Upon R. solanacearum infection (RSI), three early molecular events were possibly induced in peanuts, including Ca2+ activating CMLs/CDPKs-WRKY module to regulate the expression of resistance/susceptibility-related genes, auxin signaling was induced by AUX/IAA-ARF module to activate auxin-responsive genes that contribute to susceptibility, and MEKK1-MKK2-MPK3-WRKYs was activated by phosphorylation to induce the expression of resistance/susceptibility-related genes. Our research provides new ideas and abundant data resources to elucidate the molecular mechanism of the peanut response to R. solanacearum and to further improve the bacterial wilt resistance of peanuts.

[Analysis of syndromic surveillance of 63 325 outpatients in Fever Clinic in Beijing].

OBJECTIVE: To describe findings from syndromic surveillance of Fever Clinic visits and to determine the utility of monitoring Fever Clinic admissions as an indictor of respiratory infectious disease activity in Beijing. METHODS: A census on outpatients in Fever Clinics was conducted in two grade 3 general hospitals in Beijing from April 1, 2009 to March 31, 2010 based on a typical survey, the epidemiological characteristics of outpatients were analyzed and correlation among Fever Clinic visits, acute febrile respiratory illness (ARI) visits, influenza-like illness (ILI) visits and influenza visits determined. RESULTS: The seasonal patterns for Fever Clinic visits, ARI visits and ILI visits were quite similar, but that for influenza visits peaked later than those for ARI and ILI visits. There were high positive relationships between ARI visits, ILI visits or influenza visits and Fever Clinic visits, with a pearson's correlation coefficient of 0.99, 0.99 and 0.48, respectively (P<0.001). CONCLUSION: Syndromic surveillance of Fever Clinic visits is valuable for early warning of respiratory infectious disease outbreaks. The Fever Clinic provides a platform for early diagnosis and treatment of respiratory infectious disease.

Functional Contributions of Antigen Presenting Cells in Chronic Graft-Versus-Host Disease.

Chronic graft-versus-host disease (cGVHD) is one of the most common reasons of late non-relapse morbidity and mortality of patients with allogeneic hematopoietic stem cell transplantation (allo-HSCT). While acute GVHD is considered driven by a pathogenic T cell dominant mechanism, the pathogenesis of cGVHD is much complicated and involves participation of a variety of immune cells other than pathogenic T cells. Existing studies have revealed that antigen presenting cells (APCs) play crucial roles in the pathophysiology of cGVHD. APCs could not only present auto- and alloantigens to prime and activate pathogenic T cells, but also directly mediate the pathogenesis of cGVHD via multiple mechanisms including infiltration into tissues/organs, production of inflammatory cytokines as well as auto- and alloantibodies. The studies of this field have led to several therapies targeting different APCs with promising results. This review will focus on the important roles of APCs and their contributions in the pathophysiology of cGVHD after allo-HSCT.

Cytokine Cocktail Promotes Alveolar Macrophage Reconstitution and Functional Maturation in a Murine Model of Haploidentical Bone Marrow Transplantation.

Infectious pneumonia is one of the most common complications after bone marrow transplantation (BMT), which is considered to be associated with poor reconstitution and functional maturation of alveolar macrophages (AMs) post-transplantation. Here, we present evidence showing that lack of IL-13-secreting group 2 innate lymphoid cells (ILC2s) in the lungs may underlay poor AM reconstitution in a mouse model of haploidentical BMT (haplo-BMT). Recombinant murine IL-13 was able to potentiate monocyte-derived AM differentiation in vitro. When intranasally administered, a cocktail of granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-13, and CCL2 not only promoted donor monocyte-derived AM reconstitution in haplo-BMT-recipient mice but also enhanced the innate immunity of the recipient animals against pulmonary bacterial infection. These results provide a useful clue for a clinical strategy to prevent pulmonary bacterial infection at the early stage of recipients post-BMT.

[Analysis of disease types of outpatients with intestinal diseases from two grade-three general hospitals in Beijing].

OBJECTIVE: To understand the spectrum of diseases and epidemiological characteristics of outpatients at Entric Disease Clinic, with a focus on analysis of the distribution of infectious diarrheal diseases in different populations and to explore disease control strategies on Enteric Infectious Diseases for focal groups. METHODS: A census on outpatients at Entric Disease Clinics was conducted in two class Three comprehensive hospitals in Beijing from April 1 to October 31, 2009 based on a descriptive study using diarrhea-syndrome surveillance system set in the two clinics, thus to depict the spectrum of diseases and epidemiological characteristics of outpatients, and analyze the proportion of infectious diarrhea in diarrheal diseases specifically and the rate changes of infectious diarrhea in different months, age groups and occupational groups. RESULTS: Diseases are varied at the two enteric diseases clinics among the patients and there are mainly 10 kinds of diseases, "non-infectious diarrhea" accounted for the highest percentage (77.4%), followed by "unspecified diarrhea" (11.7%), and infectious diarrhea accounted for the least proportion(8.7%)."Gastroenteritis and enteritis" are the most frequently diagnosed cases among all the diseases, was a total of 7 565 cases, accounting for 70.2%. The volume of visits reached its top during summer and autumn(July to September), and the mean volume of visits in this period is (60.78+/-16.85) cases/day. The volume of visits has an obvious seasonal trend, and visits during July and August are the most frequent (41.82% altogether). Patients with "infectious diarrhea" had a highest ratio(5.3%) in May and lowest (1.1%) in October while patients with "bacillary dysentery" accounted for a highest ratio(8.2%) in September and lowest(3.8%) in April. Outpatients are mainly from Beijing city(61.9%), in which young and middle-aged people accounted for 73.9% in total, and student is the main occupation (28.8%). The distributions of diarrheal diseases are the same in different age groups but differ from different occupational groups. Infectious diarrhea accounted for a highest proportion(9.2%) in 18-to-44-year-old age group when using age grouping, and a highest proportion(15.2%) in restaurant service personnel when using occupation grouping. CONCLUSION: The volume of outpatients attended at general hospitals is overwhelming especially in July and August, and the major type is "non-infectious diseases", which indicates an arduous task on prevention and control of Enteric Infectious Diseases. Infectious diarrhea took up a certain amount, but the rate is not that high, which indicates possible missing diagnosis of patients with infectious diseases.Our focal groups would be young and middle-aged people and students in the city. Therefore, the need to extend the consultation hours is urgent. Meanwhile, for the main goal of surveillance and early warning on Enteric Infectious Diseases, all aspects of construction of Enteric Disease Clinic should be strengthened, such as enhance laboratory tests on pathogens, improve diagnosis level of physicians and etc.