At photonic Dirac points, electromagnetic waves are governed by the same equations as two-component massless relativistic fermions. However, photonic Dirac points are known to occur in pairs in “photonic graphene” and other similar photonic crystals, which necessitates special precautions to excite only one valley state. Systems hosting unpaired photonic Dirac points are significantly harder to realize, as they require broken time-reversal symmetry. Here, we report on the observation of an unpaired Dirac point in a planar two-dimensional photonic crystal. The structure incorporates gyromagnetic materials, which break time-reversal symmetry; the unpaired Dirac point occurs when a parity-breaking parameter is fine-tuned to a topological transition between a photonic Chern insulator and a conventional photonic insulator phase. Evidence for the unpaired Dirac point is provided by transmission and field-mapping experiments, including a demonstration of strongly non-reciprocal reflection. This unpaired Dirac point may have applications in valley filters and angular selective photonic devices. Observing the photonic analogue of an unpaired Dirac point is hard, as it requires breaking the time-reversal symmetry. Here, the authors use gyromagnetic materials to do that, and thus succeed in observing an unpaired Dirac point in a planar photonic crystal operating at microwave frequencies. It is well known that the bandstructure of the 2D material graphene3 possesses Dirac points at the two inequivalent corners of its hexagonal Brillouin zone (BZ), and similar pairs of Dirac points have been realized in optical lattices6 as well as classical acoustic9, photonic16, and plasmonic18 structures. In 2D systems preserving time-reversal symmetry (T), Dirac points necessarily occur in pairs19. The simultaneous existence of two Dirac cones, or “valleys”, implies that in order to probe features of 2D Dirac particles , special precautions must be taken to ensure that only one valley is ever excited. Depending on the system geometry, this is not always possible, because defects and interfaces can give rise to strong inter-valley scattering. The realization of systems hosting unpaired Dirac points would thus open up many interesting new experimental possibilities.In a two-dimensional (2D) bandstructure, the linear intersection of two bands at a single point in momentum space is called a Dirac point. Near such a point, the Bloch waves are governed by the same Dirac Hamiltonian that describes a two-component massless relativistic fermion3 or photonic graphene12 belong to the first type, which occur in lattices with unbroken T. These occur in pairs and are stabilized by a symmetry of the underlying lattice. The second type of Dirac point is associated with topologically protected surface states of three-dimensional (3D) topological insulators. Weak 3D topological insulators host topologically protected surface states consisting of paired Dirac points27; a photonic version of this has been demonstrated recently14. Strong 3D topological insulators, on the other hand, have topological surface states that form unpaired Dirac points30; this has been observed in condensed-matter systems30 but has never been experimentally achieved in a classical-wave system such as a photonic crystal (PhC).There are three distinct ways in which Dirac points can appear in bandstructures. Dirac points in graphene32. They correspond to the transition point at which the bulk bandgap closes. This is exemplified theoretically by the Haldane model of a 2D honeycomb lattice, where an unpaired Dirac point appears during a topological transition between the conventional insulator and Chern insulator phases32. Notably, this requires T to be broken at the transition point (the sign of the T-breaking determines which BZ corner the Dirac point occurs at). Similarly, unpaired Dirac points can also arise in Floquet systems that are driven periodically in time31. Floquet systems have been experimentally simulated using 3D photonic structures of coupled waveguides36, and a single Dirac cone has been demonstrated to exist in the “quasienergy” spectrum (where on-axis momentum plays the role of energy)34. However, many phenomena associated with unpaired Dirac points, such as one-way Klein tunneling37, are challenging to observe in that platform due to small system sizes, lack of frequency selectivity, and other technical limitations. To our knowledge, there is no previous realization of an unpaired Dirac point in the energy (rather than quasienergy) bandstructure of any 2D classical-wave system, corresponding to the closing of a bulk bandgap in a topological phase transition.The third type of Dirac point occurs at topological phase transitions between topologically trivial and nontrivial phases with broken T39. Transmission measurements reveal that the closing and re-opening of the band gaps occur at the orientation angle predicted by numerical calculations. Using direct field mapping, we show that an unpolarized point source excites Bloch states in a single valley, centered on one of the two inequivalent BZ corners. Finally, we demonstrate the phenomenon of non-reciprocal reflection: microwave beams incident from different directions outside the PhC are selectively reflected, depending on the availability of an unpaired Dirac point within the PhC to couple to. The realization of an unpaired photonic Dirac point in a PhC setting opens up many possibilities for exploring the behavior and applications of 2D Dirac modes, including valley filters40 and angular selective devices41.Here, we report on the experimental observation of an unpaired photonic Dirac point in a 2D PhC operating at microwave frequencies. Here T symmetry is broken by using gyromagnetic materials biased by an external magnetic field. The PhC is fine-tuned by varying the orientation of three dielectric scatterers in each unit cell, which controls the magnitude of parity symmetry (P) breaking. At a specific orientation angle, an unpaired Dirac point appears between the second and third transverse-magnetic (TM) polarized bands, at a corner of the hexagonal BZ (some of the methods we use to probe the unpaired Dirac point will exploit the fact that it occurs at a nonzero wave vector). The unpaired Dirac point corresponds to the transition point between a Chern insulator phase (with nonzero Chern numbers) and a topologically trivial insulator phase (with zero Chern numbers). This set of features is similar to the theoretical Haldane model in the T-broken regime a = 17.5 mm. Each unit cell comprises a gyromagnetic cylinder surrounded by three right-triangular dielectric pillars . The PhC is placed in an air-loaded parallel-plate waveguide . For θ = 0° and zero biasing magnetic field, the PhC is P and T symmetric, and its bandstructure contains two Dirac points located at the K and Κ′ points (the corners of the BZ), similar to graphene. Applying a static magnetic field B = 0.4 Tesla along the z-axis breaks T, and hence opens a complete photonic bandgap between the second and the third TM bands, as shown numerically in Fig. θ widens the bandgap at K while narrowing the bandgap at Κ′. At the specific value θ = 12.9°, the bandgap at Κ′ closes completely to form a Dirac point at 9.01 GHz. Further increasing θ, e.g. to 30°, reopens the bandgap. Figure K and Κ′ versus θ, showing the formation of unpaired Dirac points at θ = −12.9° (on K) and θ = 12.9° (on Κ′).The PhC, depicted in Fig. Κ (Κ′) isΚ (Κ′) point, K and Κ′. An unpaired Dirac point forms at K or Κ′ when θ = −12.9° and 12.9° in the actual PhC with a B = 0.4 Tesla applied magnetic field. Besides tuning 32. Three states with Chern number +1, −1, and 0 appear in the phase diagram. The unpaired Dirac point exists at the boundary between trivial and nontrivial states ; the sample configuration is shown schematically in the inset to Fig. θ = 0° and B = 0 Tesla, with the results shown in Fig. An important property of this unpaired Dirac point is that it occurs at the θ = 12.9° and B = 0.4 Tesla) from the left-upper empty region at 9.05 GHz (close to the Dirac frequency). The measured electric field pattern shows that reflection is suppressed at an angle of α = 39.1°; this is confirmed by numerical simulations  can couple to the single valley states at Κ′. Because the zigzag interface between the PhC and air preserves the valley degree of freedom44, the beam incident at α undergoes “topologically protected refraction”45, experiencing almost perfect transmission  is difficult to been achieved in similar angular selective devices.Finally, we show that the unpaired Dirac point enables the phenomenon of non-reciprocal reflection. As shown in Fig. 32 at terahertz46 or even optical frequencies47, provided suitable gyromagnetic or gyroelectric materials can be adopted. Moreover, the availability of a practical experimental platform hosting an unpaired Dirac point enables multiple avenues for follow-up explorations of the properties and uses of photonic Dirac modes. Two exemplary applications can be envisioned. The first is photonic valley filtering40: in contrast to previous time-reversal-invariant valley PhCs, which possess two opposite valleys, the current time-reversal-broken PhC has a single valley near the unpaired Dirac point, which allows for the filtering of unwanted valley signals in valley photonics. The second is non-reciprocal angular selectivity41: the current PhC is a perfect reflector for almost all incident angles, with an extremely narrow near-100% transmission window in a particular direction matching the unpaired Dirac point. This angular selectivity may be used to enhance the performance of non-reciprocal photonic devices.We have observed the experimental signatures of an unpaired photonic Dirac point in a 2D planar gyromagnetic PhC at microwave frequencies. The unpaired Dirac point occurs at the phase transition between a topologically nontrivial Chern insulator and a topologically trivial photonic insulator, as verified by transmission measurements. We confirm the existence of valley states at one corner of the BZ but not the other by direct field mapping of the microwaves refracting into an empty waveguide, as well as non-reciprocal reflection from the surface of the PhC. This work may provide useful guidelines for constructing unpaired Dirac pointsMs = 1780 Gauss, with a ferromagnetic resonance linewidth of 35 Oe. Typically, the magnetization will decrease to a negligible value without the presentence of the external magnetic field , a ferrite with measured relative permittivity 13.0 and dielectric loss tangent 0.0002. Its measured saturation magnetization is a, far exceeding practical structural dimensions (10a).The dispersion relations and field patterns are simulated using the finite element software COMSOL Multiphysics. The bandstructures in Fig. Supplementary InformationPeer Review File


Diabetic nephropathy (DN) is mainly regarded as diabetic glomerulopathy, and its progression is tightly correlated with tubular epithelial lesions. However, the underlying molecular mechanisms linking tubular damage and glomerulopathy are poorly understood.Methods: We previously reported that the upregulation of Bim mediated proximal tubular epithelial cell (PTEC) apoptosis and was crucial in the early stages of DN. Herein we modulated Bim expression in PTECs and subsequently determined podocyte (PC) cytoskeletal arrangement by building a Transwell co-culture system in high glucose (HG).Results: Compared to normal glucose, exposure to 40 mM of HG for 48 h induced significant expression of Bim in PTECs and disorganization in the PC cytoskeleton. When cocultured with PTECs in HG, exacerbated filamentous actin (F-actin) rearrangement and reduced synaptopodin levels were detected in PCs. In contrast, gene knockdown of Bim in PTECs was correlated with the absence of PC cytoskeletal disorganization. NFAT2 level and its nuclear translocation in PTECs were decreased by suppressing Bim expression. Upregulating NFAT2 disrupted the beneficial effects on F-actin organization in PCs obtained by inhibiting Bim. LncRNA microarray analysis identified NONHSAT179542.1, which was implicated in Bim-mediated PC cytoskeletal disorder.Conclusion: Our study clarified the functional role of Bim, a pro-apoptotic factor, which is involved in the crosstalk between PTECs and PCs. Bim promotes NFAT2 activation in PTECs, inducing the downregulation of lncRNA NONHSAT179542.1 in PCs, contributing to the cytoskeletal damage. Identification of the role of the Bim/NFAT2 pathway may represent a promising research direction for a better understanding of DN development. Diabetes mellitus (DM) is a growing public health problem worldwide with a high risk of severe complications Accumulating evidence has indicated that tubular damage is a key cause of chronic kidney injury The glomerulus is centrally involved in the development of DN  in vitro co-culture system to emulate in vivo environment, we have identified the underlying molecular mechanism of the crosstalk between PCs and PTECs during the early stage of DN pathogenesis.In this study, we elucidated the functional role of the Bim/NFAT2 pathway in the crosstalk between renal tubules and PCs that was involved in DN development. The early dysfunction of PCs in DN was preliminarily caused by actin remodeling abnormalities and cytoskeletal rearrangement 2 and were passaged every other day.Conditionally immortalized human PCs were donated by Dr. Yi Fan  and cultured in RPMI-1640 medium  supplemented with 10% fetal bovine serum (FBS) , 100 units/mL penicillin and 100 mg/mL streptomycin  until the cells became confluent. HK-2, a human PTEC line which had been confirmed by short tandem repeat (STR) profiling, was purchased from ProCell Corporation  and maintained in RPMI-1640 medium with 10% FBS, 100 units/mL penicillin and 100 mg/mL streptomycin. All cells were grown at 37 °C with 5% COTo determine the optimal concentration of treatment with HG and treatment time, PCs were cultured in 6-well plates for 24 h, 48 h, and 72 h in RPMI-1640 medium containing 5.5 mM glucose supplemented with 0, 10, 20, 30, 40, or 50 mM of D-glucose . Subsequently, PCs were collected for detecting the level of the cytoskeleton-related protein, synaptopodin. Mannitol was used to achieve equivalent osmotic pressure, and L-glucose  served as a control to determine the uniqueness of D-glucose.4/well in the lower chamber (a 6-well plate), which contained 40 mM HG medium; the upper chambers containing the HG medium were seeded with 5 × 104/mL control PTECs or PTECs transfected with Lenti-virus (Lenti-Bim-shRNA) as described in the “Bim Lentiviral vector construction” section below. There was communication between different cells in the system through a Polyester (PET) membrane for 24 h, 48 h, and 72 h. As controls, PCs were also cultured in 6-well plates under normal glucose (NG) and HG medium without inserts.Transwell cell-culture inserts  were placed in RPMI-1640 medium with 10% FBS and 1% antibiotics in the upper and lower compartments. The heights of the medium in the upper and lower compartments were maintained at similar levels, so the bulk flow was not due to a hydrostatic pressure gradient. PCs were resuspended at 5 ×10The experimental groups included: monoculture of PCs in NG and HG, coculture of PCs with control PTECs in HG, coculture of PCs with PTECs transfected with Lenti-Bim-shRNA (PTEC-Bim-shRNA) in HG, and coculture of PCs with PTECs transfected with negative control virus (PTEC-Bim-shNC) in HG. All co-cultures were set up in triplicate.Bim Lentiviral vector was purchased from GeneChem Corporation . Bim short hairpin RNA (shRNA) target sequence, CAC AGT TCG AGC GAT CTG TTA, was cloned into the hU6-MCS-Ubiquitin-EGFP-IRES-puromycin vector (Lenti-Bim-shRNA). The same vector was used to insert the sequence TTC TCC GAA CGT GTC ACG T as a negative control (Lenti-Bim-shNC). PTECs at 30-40% confluency were transfected with Lenti-Bim-shRNA (PTEC-Bim-shRNA) or Lenti-Bim-shNC (PTEC-Bim-shNC) according to the manufacturer's protocol . After 48 h transfection, the cells were selected with 1 μg/mL puromycin for one week. The PTEC lines with stable knockdown of Bim were established. RNA was extracted, and lysates were collected for Western blotting to confirm the efficiency of the intervention.The RNA of PTECs from different upper chambers of the Transwell co-culture system was extracted, including control PTECs, PTEC-Bim-shRNA, and PTEC-Bim-shNC. The RNA was reverse-transcribed into cDNA, which was further amplified. Quantitative real-time polymerase chain reaction (RT-PCR) was performed to detect the mRNA expression level of NFAT family members, including NFATc1, NFATc2, NFATc3, NFATc4, and NFATc5, following alteration by Bim. The subtype of NFAT, which was regulated by Bim, was determined, and Western blotting was conducted to confirm the relationship between Bim and the downstream target effector.PTEC-Bim-shRNA and PTEC-Bim-shNC as well as un-transfected controls were fixed in 4% paraformaldehyde for 30 min followed by permeabilization in 0.5% Triton X-100 for 5 min. Subsequently, cells were incubated with primary antibodies against NFAT2  at 4 °C overnight. After washing, the slides were stained with Alexa Fluor 594-conjugated secondary antibodies  and double-stained with Hoechst 33342  to visualize nuclei. Slides were rinsed twice with PBS and mounted on glass coverslips using ProLong™ Glass Antifade Mountant . Cells were observed, and images were captured at randomly selected fields using an LSM780 confocal microscope .The PTEC-Bim-shRNA were transiently transfected with NFATc1  or control empty plasmids. Briefly, Opti-MEM medium , containing 2.5 μg p-NFATc1 or empty plasmids, was mixed with 5 μL lipofectamine 3000 transfection reagent  and the mixture was added to the cells. After 8-12 h, the Opti-MEM medium was removed, and the cells were incubated with RPMI-1640 medium containing 10% FBS. Forty-eight hours after the transfection, plasmid transfection efficiency was evaluated by RT-PCR and Western blotting.The siRNAs against NFATc1 were transfected into control PTECs using lipofectamine 2000 transfection reagent  by following the manufacturer's instructions. The siRNAs against NFATc1 (siNFATc1) were synthesized by Shanghai GenePharma. Sense sequences for NFATc1 siRNAs were 5'-GCA UGG CUA CUU GGA GAA UTT-3' (siNFATc1-1) and 5'-AUU CUC CAA GUA GCC AUG CTT-3' (siNFATc1-2). Sense sequences for the negative control were 5'-UUC UCC GAA CGU GUC ACG UTT-3' (siNC-1) and 5'-ACG UGA CAC GUU CGG AGA ATT-3' (siNC-2). The siRNA knockdown efficiency was verified by RT-PCR and Western blotting. Thus, PTECs with stable knockup (p-NFATc1) and knockdown of NFATc1 (siNFATc1) were established. Each transfection was repeated in triplicate. After successful transfection, the cells were seeded in the upper chamber of the Transwell system and were cocultured with PCs in HG condition by following the instructions described.After monoculture of PCs and coculturing with various PTECs in which Bim expression was regulated by various treatments in HG for specific periods , synaptopodin expression in PCs was detected as described in the “Immunofluorescence (IF) staining” and “Western blotting” section below.After the downstream target of Bim in PTECs was verified, synaptopodin expression in various PCs was examined as follows: PC groups cocultured with PTECs carrying various vectors including, siNFATc1, siNC, Lenti-Bim-shRNA, Lenti-Bim-shRNA carrying p-NFATc1, and Lenti-Bim-shRNA carrying empty plasmid as well as control PTECs.PCs were seeded on sterile glass coverslips in the lower chamber of the Transwell system or a sole 6-well plate and cultured under various conditions. Subsequently, glass coverslips with PCs were extracted to detect IF intensity. The slides were fixed for 30 min with 4% paraformaldehyde, followed by permeabilization with 0.5% Triton X-100 in PBS for 5 min. Thereafter, the slides were incubated with the TRITC Phalloidin  or synaptopodin antibody  at 4 °C overnight. Next day, Alexa Fluor 594-conjugated secondary antibodies , along with DAPI stain to visualize the nuclei, were applied to detect synaptopodin. Images were taken at randomly selected fields using an OLYMPUS FSX100 imaging system .After the regulation of NFAT2, IF staining for synaptopodin and F-actin fiber arrangement in PCs was also performed in a similar manner as delineated above.-ΔΔCT method was used to calculate the relative expression levels of each mRNA. RT-PCR was performed in triplicate.Total RNA was extracted from the cultured cells using TaKaRa RNAiso Plus (Cat.#9108) following the manufacturer's protocol, and the concentration of the total RNA was quantified by measuring the absorbance at 260 nm. RNA from each sample (1 μg) was added to 20 μL reaction mixture, and cDNA was synthesized using the PrimeScript™ RT reagent Kit with genomic DNA Eraser . Quantitative RT-PCR was performed using UltraSYBR Mixture (low ROX)  to detect Bim, NFATc1, NFATc2, NFATc3, NFATc4 and NFATc5 mRNA expression. β-actin was used as an internal control. The primer sequences are shown in Proteins were isolated from various cultured PTECs and PCs, and protein levels were detected. Briefly, protein extracts were boiled in RIPA buffer  and separated by SDS-PAGE electrophoresis. Western blotting was performed with the antibodies against synaptopodin (syn)  to detect its protein level in PCs. Anti-Bim antibodies  were used to examine the HG-induced Bim expression and the inhibition effect on PTECs after transfection with the Lentiviral vector. Antibodies against NFAT2  were used to detect NFAT2 levels after transfection with siNFATc1 or p-NFATc1 into PTECs or PTEC-Bim-shRNA. Anti-MICAL2 antibodies  were used to detect the MICAL2 level in PCs after coculturing with un-transfected PTECs and PTEC-Bim-shRNA, and after transfection of NONHSAT179542.1 siRNA and siNC into PCs. β-actin  was used as a reference protein.Total RNA was isolated using TaKaRa RNAiso Plus (Cat. #9108) according to the manufacturer's instructions. Total RNA was assessed for a RIN number using Agilent Bioanalyzer 2100 . The biotinylated cRNA targets for the Sino-Human lncRNA array V3.0. were generated by RNA samples of each group and then were hybridized with probe on the slides. Upon hybridization, slides were scanned on the Agilent Microarray Scanner . Data extraction was conducted with Feature Extraction software 10.7 . The Quantile algorithm of R package “limma” was used to normalize raw data. The microarray experiments and data analysis were performed by Shanghai Sinomics Corporation  following the protocol of Agilent technologies Inc at Sinotech Genomics Corporation. The lncRNAs were constructed using the public transcriptome databases . The microarray also included an entire collection of 25353 protein-coding mRNAs. Genes with a fold change of at least 1.5 were selected for further analysis. Heatmap was generated by an R package “pheatmap”.Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis was performed to identify significant pathways. Target gene prediction of lncRNAs by cis/trans was conducted. Cis target gene prediction was to seek target mRNA located near lncRNA within 10 kb. Trans target gene prediction was carried out following the principle of sequence complementary pairing. Complementary mRNA with lncRNA was acquired by blast alignment. We used RNAplex software to calculate the thermodynamic parameters of lncRNA complementary with mRNA, and the sequence e ≤30 was extracted. The target network was constructed using Cytoscape.Expression of fourteen lncRNAs was validated by RT-PCR. The cDNA was synthesized by using a PrimeScript™ RT reagent kit (TaKaRa). Then, RT-PCR was performed using UltraSYBR Mixture (low ROX) . Primers for NONHSAT253617.1, XR_929282.2, NONHSAT179542.1, NONHSAT251382.2, XR_934942.2, NONHSAT220341.1, ENST00000542022, NONHSAT040129.2, NONHSAT227535.1, NONHSAT203700.1, NONHSAT243847.1, NONHSAT040287.2, NONHSAT179858.1, and NONHSAT179237.1 were synthesized by RiBo-Bio . All RT-PCR primer sequences are shown in After validation, the sequence of siRNA against NONHSAT179542.1 was designed by RiBo-Bio . The sequence of siRNA against NONHSAT179542.1 was as follows: 5'- ACTTGGAAACTGCCAGAAA-3'. PCs after coculturing with PTEC-Bim-shRNA were transfected with 50 nM NONHSAT179542.1 siRNA or siNC using Lipofectamine™ 2000 (Invitrogen) according to the manufacturer's protocols. After transfection for 48 h, the RNA was extracted and the intervention effect was validated by RT-PCR.To determine the influence of NONHSAT179542.1 on the PC cytoskeleton organization, IF staining for F-actin and synaptopodin was conducted. Synaptopodin expression in PCs after downregulation of NONHSAT179542.1 was examined by Western blotting.The groups involved cocultures of control PTECs, PTEC-Bim-shRNA, and PTEC-Bim-shNC with PCs. Also, PTEC-Bim-shRNA were cocultured with PCs transfected with NONHSAT179542.1 siRNA (PTEC-Bim-shRNA + PC-lncRNA-siRNA), or with NONHSAT179542.1 siNC (PTEC-Bim-shRNA + PC-lncRNA-siNC). All co-cultures were set up in triplicate.P < 0.05.All statistical analyses were performed using SPSS Statistics 22.0 . A Student's t-test was used to assess significance for data within two groups. Multiple statistic comparisons were analyzed using one-way ANOVA, followed by post hoc tests. Data were presented as mean ± standard error of the mean (S.E.M.), and the level of statistical significance was estimated at P < 0.05); other glucose concentrations showed no significant effect. HG treatment of D-glucose for 24 h and 72 h had no significant difference in the synaptopodin level compared with the normal group . To investigate the role of increased Bim in PCs cytoskeletal organization, Bim expression was further regulated. The expressions of Bim in PTECs after alteration were investigated by RT-PCR and Western blotting analysis. Compared with the vehicle control, Bim expression was significantly decreased in cultured PTECs (P < 0.05) following transfection with Lenti-Bim-shRNA , which inhibited Bim expression in PTECs . Results of Western blotting analysis were consistent with RT-PCR and NFAT2 expression was found to be regulated by Bim in PTECs .To confirm Bim-mediated PC cytoskeletal disarrangement via NFAT2, we regulated Bim and NFAT2 expression in PTECs and determined F-actin arrangement and synaptopodin level in PCs. RT-PCR and Western blotting analyses revealed that PTECs after transfection with siNFATc1 showed downregulation of NFAT2 compared with control PTECs . Variations of lncRNA expression from the lncRNA microarray data are shown in Figure To systematically identify lncRNAs in PCs after various treatments, we compared differential lncRNA expression profile by microarray analysis of PCs between the two cocultured groups (PTEC-Bim-shRNA vs control PTECs). A heatmap between two groups revealed differentially expressed lncRNAs with statistical significance  have been shown to correlate biomarkers of PC damage with proximal tubule dysfunction, raising the possibility of an AGE-mediated impact on both glomeruli and proximal tubules Besides pro-apoptosis, the additional regulatory role of Bim in inducing cytokine production via activation of the nuclear factor of activated T cells (NFAT) was clarified In the present study, we analyzed the expression changes of all five NFAT family subtypes in PTECs, and identified NFAT2 as a potential downstream target of Bim. NFAT2 functions as a transcription factor and is involved in gene regulation by binding to specific gene promoters, requiring its translocation to the nucleus. We observed increased nuclear levels of NFAT2 in HG-treated PTECs that overexpressed Bim, which was highly suggestive of nuclear translocation of NFAT2. On the contrary, low levels of Bim led to reduced nuclear translocation of NFAT2, again indicating Bim-induced NFAT2 nuclear translocation. These findings indicated that the downstream target of Bim, NFAT2, might be important in mediating the cytoskeletal disorganization in PCs. Further studies revealed that inhibition of NFAT2 abrogated cytoskeletal disruption in PCs after coculturing with PTECs, while up-regulation of NFAT2 induced synaptopodin reduction and cytoskeletal rearrangement in PCs despite suppression of Bim. These results revealed NFAT2's role as a potential target of Bim involved in the cytoskeletal damage of PCs.2+-dependent remodeling of the actin cytoskeleton in PCs was shown to be essential to counteract mechanical forces in response to mechanical load in vitro and mice resistant to synaptopodin degradation were protected from proteinuria in vivoPreviously, CaAn important role of hyperglycemia in abnormal activation of T cells has been described previously To further explore the molecular mechanisms involved in the cytoskeletal disruption in PCs, we performed lncRNA microarray analysis of PCs cocultured with PTECs after the regulation of Bim. LncRNAs, with sequence length greater than 200 nucleotides, have been suggested to regulate the gene expression during normal development and diseases, including DM and other complex disorders To address this issue, we used Sino-human lncRNA array V3.0 to detect the differentially expressed lncRNA profile in PCs after coculturing with PTECs. Several differentially expressed lncRNAs were selected for further verification by RT-PCR, identifying NONHSAT179542.1, NONHSAT040129.2, and NONHSAT203700.1 for subsequent studies. Based on KEGG functional annotation data and target gene prediction of lncRNA microarray analysis of PCs, NONHSAT179542.1 was chosen for further analysis. Functional studies showed that inhibition of NONHSAT179542.1 in PCs markedly exacerbated the cytoskeletal damage which was alleviated by suppression of Bim in PTECs, indicating NONHSAT179542.1's role in Bim-mediated PC cytoskeletal reorganization in DN. Downstream mechanism of NONHSAT179542.1 was further explored and 218 genes were identified as the potential targets of NONHSAT179542.1 by cis/trans target gene prediction. By microarray data analysis and the NONHSAT179542.1 regulation study, MICAL2, a flavoenzyme that binds to F-actin and triggers its depolymerization through redox modification In this study, we have provided evidence that HG-induced Bim overexpression in PTECs exacerbated PC cytoskeletal disruption by targeting NFAT2 and the key downstream targets, lncRNA NONHSAT179542.1/MICAL2 were implicated in it. Our data clarified insightful molecular mechanisms of tubule-PC crosstalk as a potential approach in DN treatment.Supplementary figures and tables.Click here for additional data file.


Malnutrition and MR could be reduced by increasing economic level, supporting people living alone or being single, widowed, and divorced, and improving lifelong learning.Malnutrition in the elderly could be tackled by addressing socioeconomic factors. This study aimed to determine the magnitude of the relationship between socioeconomic factors and the malnutrition or malnutrition risk (MR) in the elderly. The PubMed and SCOPUS databases were searched for observational studies that included assessment of malnutrition or/and MR and socioeconomic variables  in ≥60-year-old subjects, published in English among 2000–2018 (PROSPERO: CRD42019137097). The systematic review included 40 observational studies  and 16 cross-sectional studies in the meta-analysis  of malnutrition and MR in relation to low educational level (Odds Ratio (OR): 1.48; 95% Confidence Interval (CI): 1.33–1.64;  Nowadays, the aging of the population is a consequence of the increase in life expectancy . In Euro2 or by meeting two of these three criteria: unintentional weight loss (>10% in an indefinite time period or >5% over the last three months) combined with either a low BMI  or a low fat-free mass index (FFMI) score  [Aging can be associated with malnutrition, which is a public health problem characterized by a multifactorial physiological state ,4. Malnuctively) . In addictively) . As a reHowever, before reaching the need of a malnutrition diagnosis, the use of validated tools could be used to determine malnutrition risk ,7. AccorConsequently, a state of malnutrition can cause an impairment of quality of life, especially in the elderly, and can lead to increased healthcare costs and hospital stays .Additionally, malnutrition is related to health, environmental, and social factors or determinants . The eldThus, the hypothesis was that social and economic factors—such as low educational level, living alone, being single, widowed, or divorced, low income level, low previous (before 60 years old) or current occupational level, feeling of loneliness, living in rural areas, and low food expenditure—are related to malnutrition and malnutrition risk in the elderly.The main objective of the present systematic review and meta-analysis was to assess and determine the magnitude of the relationship between socioeconomic factors and the malnutrition or malnutrition risk in the elderly ≥60 years old.We performed a systematic review and meta-analysis of observational studies regarding malnutrition and malnutrition risk and their relation to socioeconomic factors in the elderly. This study was reported in accordance with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA)  and it wAn exhaustive literature search in the PubMed and SCOPUS electronic databases was carried out, as these databases are considered the largest and the most multidisciplinary, covering medical and social sciences journals that include the principal outcomes of the present meta-analysis . Depending on the study, socioeconomic factors were assessed separately as social factors and economic factors. Regarding nutritional status, some studies assessed the malnutrition and the malnutrition risk separately, while other studies evaluated them together. For this reason, the search was done using the following keywords combined with each other: malnutrition, nutritional status, older, elderly, social factors, social, economic, and socioeconomic. The search strategies in PubMed were: “older” or “elderly” and “social” and “malnutrition”, “older” or “elderly” and “economic” and “malnutrition”, “nutritional status” and “social factors”; and in SCOPUS were: “economic” and “malnutrition” and “older” or “elderly”, “social” and “malnutrition” and “older” or “elderly”.The inclusion criteria were observational studies , with the target population ≥60 years old, in which publications with data on malnutrition status, as well as social and economic variables, were published in English between January 2000 and December 2018.The studies that did not meet all of the inclusion criteria were excluded. Additionally, studies that only included populations with illness or comorbidities were also excluded in order to focus on the general population, who was not selected taking into account a specific disease, to be able to generalize the results.www.covidence.org). Firstly, the studies were selected based on the title information. Subsequently, the abstracts of these selected studies were checked to see if they met or not the inclusion and exclusion criteria. Finally, those studies that met the inclusion criteria were full-text assessed. When the studies did not have all of the information necessary to be included in the present study, an email was sent to the authors of the publication requesting this information. The data extraction was carried out independently. Of the included studies, the following variables were collected: authors, year of study development, the country of implementation, the type of study, number of participants, target population’s age, gender, social factors , economic factors , nutritional status screening tool, nutritional status , and study quality. The previous or current occupational level was divided into low occupational levels  and high occupational levels .The search strategy of the studies that matched the inclusion criteria was carried out with the online platform Covidence  . To select studies for the meta-analysis, it was taken into account whether they used the MNA as a nutritional evaluation tool  and evaluated the socioeconomic variable with the same criteria. To include a study, information about the percentage or number of subjects with a risk of malnutrition or malnutrition-related to the socioeconomic variable was required. In contrast, studies that assessed all outcomes via multivariate analysis were excluded from the meta-analysis. Thus, we only performed the meta-analysis of each socioeconomic factor, in which at least 4 studies evaluated this factor and followed the inclusion criteria described previously. In addition, an analysis was performed in general  and by a subgroup of the same variables. To evaluate the heterogeneity of the studies, we used the In = 48), text not available (n = 8), non-representative population sample (n = 5), and wrong study design (n = 5). Finally, as shown in A total of 363 studies were obtained from the literature search, of which 60 were eliminated by duplicity. Of the remaining 303 studies, 197 were excluded according to the inclusion and exclusion criteria based on the information provided in the title and the abstract. A total of 106 studies were full-text assessed, and 66 were excluded because of the following reasons: results not related to the outcomes studied n = , text noOf the total 40 included studies, 36 were cross-sectional ,51,52,53Of the 40 observational studies included in the systematic review, regarding the tools used to assess the nutritional status, 29 studies used the MNA ,51,52,53Of the total of 40 observational studies included in the systematic review, eight different socioeconomic variables were evaluated: 31 studies evaluated the educational level ,51,52,53Of the 31 studies that evaluated educational level, 15 established significant relationships with regard to malnutrition or malnutrition risk ,49,52,53A total of 27 studies evaluated the relationship between living alone or cohabiting and the malnutrition or malnutrition risk, of which 13 showed statistically significant results ,50,51,53A total of 24 studies evaluated the relation between the marital status and malnutrition or malnutrition risk, of which 11 showed significant results ,42,45,53Of the total of 20 studies that evaluated the income level, 13 established significant relationships ,45,47,51Of the total of 12 studies that evaluated the previous (<60 years old) or current occupational level, six obtained statistically significant results ,45,46,53Of the total six studies that evaluated the feeling of loneliness, four were related significantly with malnutrition or malnutrition risk ,17,21,33A total of three studies evaluated the place of residence, which was differentiated between urban and rural. Only one study established significant relationships with malnutrition or malnutrition risk  and two Only one study evaluated the relation between food expenditure and malnutrition or malnutrition risk, and it did not show statistically significant results  Table S.Of the total 40 studies included, 16 were of high quality ,48,51,53p < 0.001; I2 = 94%; p about heterogeneity < 0.001)  . The 16 p < 0.001; I2 = 91%; p about heterogeneity < 0.001)  .p < 0.001; I2 = 90%; p about heterogeneity < 0.001)  .p < 0.001; I2 = 91%; p about heterogeneity < 0.001)  .p < 0.001; I2 = 89%; p about heterogeneity < 0.001)  .The present systematic review and meta-analysis of observational studies, which were mainly cross-sectional, supports the hypothesis that social and economic factors are related to malnutrition and malnutrition risk in people aged ≥ 60 years. Focused on the meta-analysis outcomes, according to the socioeconomic factor, in a descendant order, low income level, living alone, being single, widowed, or divorced, and low educational level had a greater relationship with malnutrition and malnutrition risk in the elderly. Meanwhile, there were not enough articles providing scientific evidence to make a meta-analysis of some of the other socioeconomic factors, such as low occupational level , feeling of loneliness, living in rural areas, and food expenditure. Thus, it was not possible to prove whether these factors act as a malnutrition risk factor in the elderly.In the current meta-analysis, it was observed that low educational level is a risk factor for malnutrition, suggesting the predisposition to malnutrition. A possible explanation is that individuals with a higher educational level have more knowledge about nutrition and health , leadingAnother identified risk factor from the results of the present meta-analysis is living alone, which showed a significant association with the risk of developing malnutrition. The elderly who lived alone had a greater malnutrition risk compared to others who cohabit with someone—either family, acquaintances, or friends. Three literary reviews ,57,58 obIn reference to marital status, the present meta-analysis established an association between being single, widowed, and divorced and the risk of malnutrition. In the same way to the present results, one review concluded that widowed individuals, independently of gender, were more vulnerable to the risk of nutritional deficiencies . The posFurthermore, the present meta-analysis also established an association between lowest income levels and a higher risk of developing malnutrition in the elderly. One explanation is based on the fact that the healthiest or freshest food is the most expensive, so are not accessible for the elderly with few economic resources . Thus, aAnother socioeconomic factor evaluated is the low previous (<60 years old) or current occupational level, namely, farmer, breeder, housewife, laborer, hard manual or physical work, or unemployment, which were identified, in the present systematic review, as a malnutrition risk factor. However, there were not enough articles to perform a meta-analysis. A possible explanation is that those elderly with a low occupational level usually have a lower economic level too, so they might not have access to high-quality nutritional food.Regarding the feeling of loneliness, which is separate from social isolation, the present systematic review obtained low numbers of articles relative to the risk of malnutrition. Only two cross-sectional studies showed that the elderly with a greater feeling of loneliness were at greater risk of malnutrition ,33, whilThe present meta-analysis could not establish a reliable association regarding the place of residence, either urban or rural, and malnutrition risk in the elderly. Of all of the studies included in the review, only one cross-sectional study showed that people in Iran who lived in rural areas had a greater risk of malnutrition . In the This systematic review and meta-analysis presents different limitations. First of all, the present systematic review was about observational studies, mainly cross-sectionals. Future studies should evaluate the influence of socioeconomic factors in malnutrition of randomized controlled intervention studies to obtain maximum evidence . The secAll of the results of the present systematic review and meta-analysis serve to better understand the social and economic risk factors associated with malnutrition in the elderly ≥60 years old. Moreover, the identification of social and economic risk factors allows us to avoid them, and thus to reduce or prevent malnutrition or/and malnutrition risk and to maintain a good nutritional status of the elderly. Thus, regarding the evidence provided in the present systematic review and meta-analysis, the following table shows a proposal of actions on socioeconomic risk factors to reduce or prevent malnutrition and malnutrition risk. These actions can provide a basis for future nutritional interventions on socioeconomic factors to diminish malnutrition in the elderly. However, more research is needed to confirm that these interventions are effective in combating each socioeconomic factor .Malnutrition and malnutrition risk could be reduced in the elderly by increasing their economic level, supporting those living alone or who are single, widowed, or divorced, and improving lifelong learning.


On the eve of independence in 1962, malnutrition was the largest single cause of death in Jamaica for children under one. Although child malnutrition rates have rapidly declined since 1962, today Jamaica experiences a double burden of malnutrition: the coexistence of pockets of high child malnutrition with rising levels of childhood obesity. Based on a wide range of sources, including public documents, newspaper reports, scientific studies and reports by international agencies, this article examines a gradual decline in child malnutrition and the rise of the double burden of malnutrition in Jamaica from independence to the present. It will first of all show that changes in the global economy and overseas loans and aid both aided and limited the ability of the Jamaican government to lower child malnutrition levels and also contributed to a rise in childhood obesity. Second, it will illustrate that a traditional deficit-led approach to child malnutrition was followed in post-independent Jamaica, focussing on the public and individuals as targets for intervention and using quantitative measures to trace progress. And third, it will question whether the double burden of child malnutrition will give rise to ‘healthy publics’—‘dynamic collectives of people, ideas and environments that can enable health and well-being’. On the eve of independence in 1962, malnutrition was the largest single cause of death in Jamaica for children under one , p. 6. AThis study examines the gradual decline in child malnutrition and the emergence of the ‘double burden of malnutrition’ in Jamaica from independence in 1962 to the present. Child malnutrition refers here to undernutrition and includes: stunting, low height for age; wasting, low weight for height; and underweight, low weight for age. During the period under consideration, a range of measuring methods and classification systems were used to assess child malnutrition. While the World Health Organization (WHO) and other organisations now use the term child malnutrition for children under the age of 5, studies on child malnutrition in Jamaica after 1962 measured a range of age groups, including children enrolled in primary school. Also some key interventions adopted focussed on older children, including school feeding programmes. For that reason, this study focusses not just on children under 5 but also older children.The Government of Jamaica (GoJ), with the support of international donors, aid agencies and others, tried in various ways to lower child malnutrition rates as it realised that high levels posed a threat to the future of the island because children who suffer from malnutrition are in a critical stage of growth and if they do not catch up, are more likely not to do well in school and hence can be trapped in a cycle of poverty. And today, it undertakes efforts to lower childhood obesity rates because obese children run the risk of developing non-communicable diseases (NCDs) later in life, which in turn will place large burdens on the public health system, economy, and society. The following will show that changes in the global economy and conditions imposed by international lenders significantly affected the GoJ’s ability to lower child malnutrition levels and also contributed to a rise in childhood obesity. But it will also highlight that without international loans and overseas aid, the island would not have witnessed a gradual decline in child malnutrition in the decades following independence. In other words, it will argue that external factors, in complex interplay with local factors, acted as a double-edged sword.1 and is divided into five sections. The first provides historical background and shows that it was not until the interwar years before child malnutrition became a problem in Jamaica and surveys were undertaken to measure it. It also sets out the historical factors that gave rise to this problem and attempts made from the Second World War onwards to solve it. The second section zooms in on the years immediately leading up to and following independence and illustrates that methods pioneered in developed countries were increasingly replaced by interventions deemed more realistic within the local context, and that various international organisations, including the United States Agency for International Development (USAID) and the WHO’s regional office in the Americas—the Pan-American Health Organisation (PAHO)—offered crucial help that supplemented local efforts. In the 1970s, some of this aid declined at a time when Jamaica witnessed a nutrition crisis. The third section explains why child malnutrition levels did not increase during these challenging years, while the fourth examines how the GoJ tried to further lower child malnutrition in the 1980s and 1990s, amidst conditionalities imposed by international lenders that significantly affected household food security. And the last section traces the rise of the double burden of child malnutrition and mentions some methods adopted to address it.This study is based on a range of primary sources, including scientific studies, newspaper reports, public documents, and reports by international donors,This article will show that throughout the post-independence period, a mostly traditional deficit-led approach to child malnutrition was adopted in Jamaica; that is, focussing on the public and individuals as targets for intervention and using quantitative measures to trace progress. For instance, the GoJ ran campaigns to increase exclusive breastfeeding, set up nutrition clinics that monitored the weight of children, handed out supplementary foods for pregnant and nursing women and undernourished children, and employed public health workers who taught women how to prepare nutritious weaning foods. The conclusion will question whether it is possible for Jamaica to create what By examining the process by which child malnutrition rates declined and a double burden of malnutrition emerged in Jamaica after 1962, this article will contribute to several (overlapping) sets of scholarship. Firstly, it will add to the slowly growing body of work on health and medicine in the Caribbean. Despite its medical and cultural importance, nutrition has often been a marginal topic in Caribbean historiography, being only briefly addressed in discussions of diseases directly or indirectly linked to diet, such as tuberculosis or hookworm e.g. . In centThirdly, this study will contribute to scholarship on the history of nutrition in colonial and post-colonial contexts more generally. This body of work has focussed mostly on the first half of the 20th century. Various studies have examined attempts by colonial governments to alleviate hunger in the years before the Second World War and the science underpinning them, focussing particularly on African colonies e.g. . And somMost of this recent body of work focusses on Africa and is particularly concerned with the role of nutritional science and humanitarian actors in approaches to and discussions around child malnutrition. This case study, which pays less attention to nutritional science, will show that approaches to child malnutrition in Jamaica mirrored in many ways those adopted in former African colonies, including the reliance on overseas aid agencies and international lenders. Yet during the period under consideration, Jamaica never experienced (the threat of) famine and because it was more urbanised and a Small Island Developing State (SID), making it more reliant on global trade networks, it experienced the ‘nutrition transition’—a shift towards consuming more energy-dense processed foods and becoming less active, resulting in obesity and obesity-related NCDs—earlier than many former African colonies so that by the turn of the 21st century already a significant proportion of children were overweight or obese. Although now becoming more prevalent amongst children, in most former African colonies today obesity is still largely an adult phenomenon . This caChild malnutrition was studied and problematised long before Jamaica gained independence and was part of broader regional and imperial-wide investigations into nutrition. For instance, in 1937 a local nutrition committee was set up that measured the nutritional status of the population, which fed into the report of the Committee on Nutrition in the Colonial Empire . This coLike other plantation economies in the Caribbean, Jamaica had long been reliant on the importation of large quantities of food, a dependency that dated back to the formation of sugar plantations in the 17th century. Largely as a means to reduce costs, sugar planters reserved some land on their estates for slaves to cultivate their own food and supplemented their slaves’ diet with imported foods, such as salted, dried fish. The land allocated to slaves was usually the least fertile land, and slaves were only able to grow staple foods, such as yams, the surplus of which they were allowed to sell on slave markets. Thus during slavery, as These arrangements continued to structure food production and consumption in Jamaica after emancipation in 1838. Despite many ex-slaves seeking to become farmers, they struggled to obtain land. Various government schemes were created to facilitate an increase in peasant ownership and cultivation by the end of the nineteenth century, but most purchases under these programmes were small-scale (under 10 acres) and encompassed poor quality, hard to reach land , p. 170.Changes in demographic and economic circumstances made it even more difficult to secure adequate nutrition after the First World War. Between 1921 and 1943, the population increased by 1.6 per cent per annum , p. 135 In the 1950s, child malnutrition in Jamaica became defined as a medical-nutritional problem. Although local health researchers, centring around the Tropical Metabolism Research Unit (TMRU) at the University College of the West Indies, agreed that child malnutrition was largely the result of poverty, they did not assess the extent to which household income and other socio-economic factors contributed to child malnutrition . InsteadThe GoJ and various UN agencies operating in the island in the 1950s equally medicalised child malnutrition. The GoJ adopted various schemes largely based on child feeding practices used in developed countries, such as cod liver oil distribution schemes , p. 133,The WHO, for instance, questioned the suitability of cod liver oil distribution schemes in tropical regions where sunlight exposure throughout the year allowed for optimal levels of Vitamin D, and started issuing guidelines for large-scale nutrition surveys that included attention to social and cultural factors .2 for instance, reinforced a racialised view of the ‘normal’ and ‘abnormal’; it divided malnutrition into mild, moderate and severe forms, but used children in Boston, measured between 1930 and 1956 and of mostly European descent, for its reference population  colonies in the 1960s. The use of nutrition anthropometry in the colonies was not without its problems. The use of the so-called Gomez scale,pulation , p. 774.pulation  p. 146. The first survey undertaken in Jamaica after independence was carried out in 1963 by the nutrition unit of the Jamaican Scientific Research Council. It looked at the food intake of 665 pre-school children from 369 families in 59 different areas. Another survey took place in 1970, carried out with support from the Caribbean Food and Nutrition Institution (CFNI), a specialist centre of PAHO set up in 1967. And a PAHO study into nutritional problems in the Caribbean from 1963 to 1964 also included Jamaica . These vThe various Jamaican nutrition studies concluded that the high rate of infant deaths caused by malnutrition was the result of various socio-economic and cultural factors, most notably poverty, mirroring similar findings in other British (former) colonies , p. 239.Early and exclusive breastfeeding has long been seen as a means to enhance the survival chances of infants; breastmilk is easily digested and uncontaminated with infectious organisms so it protects against gastroenteritis. The various studies carried out in Jamaica in the 1960s found a rapid decline in breastfeeding: few mothers breastfed beyond 3 months and the trend was towards less breastfeeding and earlier introduction of the bottle see . Social Like women in other tropical countries, Jamaican women found it difficult to sterilise bottles and teats (many only had one bottle and not a separate pan to sterilise it) and because formula was expensive, they also tried to stretch it so that the baby was given diluted milk. One study estimated that it would cost half a family’s weekly income to feed a child formula according to the guidelines on the tin . Mothers who could not afford to buy expensive formula often used condensed milk, which was cheap but high in sugar. By the late 1960s, more women started to use dried skimmed milk, which contained more protein than whole milk and was cheaper , p. 146.Jamaican mothers’ assumptions about appropriate weaning foods was also singled out in various studies as a factor accounting for high child malnutrition rates. For example, mothers rarely gave children under two fish, meat or sweet potatoes as they thought that these foods produced worms. The high status attached to commercially produced weaning foods also meant that they forewent bananas and other local foods for weaning mixtures , p. 29. It was strongly recommended that infant feeding was taught through practical demonstrations, using foods, cooking equipment and methods that were feasible in local homes . Such teGradually, the teaching of infant feeding did become more ‘realistic’, ‘affordable’ and ‘local’ as it did in many other developing countries, such as Uganda . PAHO 1. For insRecognising that poverty was a main cause of child malnutrition, it was also recommended that more and better day-care facilities be set up for children of working mothers , p. 79. State interventions in this period were also stimulated by international agencies and encompassed a mix of technical and structural solutions. For instance, 3 Expanding its efforts, the GoJ began to control the price of some 15 food items to protect low-income groups, pursued a policy of import controls to encourage the use and production of local foods stuffs, ordered research into ‘indigenous agricultural produce’ that could be marketed locally, and used radio to convey messages about healthy child nutrition  government led by Michael Manley, which adopted a democratic socialist programme that included policies to reduce poverty and social inequalities. As suggested by various UN agencies, a year after it took office, it adopted a nutrition programme that addressed nutrition from different angles—health, agriculture, and education—and focussed on food supply, food demand, and the ‘biological use of food’  .The import-substitution scheme Growing and Reaping our Wealth (GROW), which was launched in 1973, was mostly a response to a rising bill of imported food stuffs, caused largely by devaluation. The food bill increased from Jamaican dollars (J$) 43.3 million in 1968 to J$60.2 million in 1971 and rose even further after the oil crisis . In factoverall improvement in the nutritional status of young children during the 1970s. First, a rapid increase in the informal sector—jobs not recognised as normal income and on which no taxes are paid, including illegal activities—provided unemployed or underemployed communities with some income. Second, policies adopted by the PNP government led to better income and employment, especially for rural households. Particularly important here was Project Land Lease under which privately held lands were leased by the government, and then (for period of 5 years) to small farmers in the area to allow them to supplement their own holdings. By 1980, almost 38,000 farmers had been placed on 75,000 acres of land . This use of lay health workers first began in 1969 when the GoJ, the University of the West Indies and Cornell University Medical College set up a rural health project in Elderslie, in the parish of St. Elisabeth, which had a high rate of infant mortality caused by malnutrition , p. 1168plements . This moThe impact of the increased use of CHAs on malnutrition varied from parish to parish. In Elderslie, for instance, it did not reduce overall levels of malnutrition but succeeded in lowering high mortality associated with nutritional deficiency because children that were mildly or moderately malnourished were sought out and then treated in their homes. Yet in Hanover parish, malnutrition levels of children under 2 fell by about 40 per cent after the introduction of CHAs. This was largely because the parish employed nearly twice as many CHAs as other parishes and also had a relatively continuous supply of food aid from the US . In 1983, 63.2 per cent of all new mothers exclusively breastfeed for the first 6 weeks , financed amongst others by USAID and the UN World Food Programme to the tune of J$141 million, was adopted that consisted of two key elements. The first was a food stamp programme for most-at-risk groups, including pregnant and nursing women, all children under 5 registered at a health centre, and some moderately to severely malnourished children. The stamps entitled them to buy powdered skimmed milk, cornmeal, and rice. In 1988, some 23 per cent of all households and 38 per cent of children under 5 received food stamps. The second element was a school feeding programme under which some 600,000 children received the nutribun or a cooked lunch either for free or a small fee . As in t10 For example, only 34 per cent of food subsidies went to the poorest 40 per cent of the population but stamps went to 57 per cent of the poorest 40 per cent. Yet only half of households with malnourished children received food stamps, largely because of problems with registration and administration of the programme , for instance, sponsored a project to set up backyard gardens in two urban communities, while Food for the Poor ran breakfast programmes for adults and children, gave fishermen in some communities equipment, ran a soup kitchen and set up chicken farms , Mother’s (1981) and Island Grill (1991). A reduction in corporate tax from 45 per cent in 1986 to 33 per cent in 2006 and other fiscal incentives that the GoJ was encouraged to adopt by the international lenders go some way to explain the rise in locally owned fast-food outlets and also the growth of local food producers and distributors like Wisynco and Grace Kennedy . BetweenNumerous studies have tried to assess the link between trade liberalisation and the prevalence of malnutrition, overweight, and obesity. A recent literature review of quantitative studies has concluded that in low-income and middle-income countries trade openness has reduced malnutrition most likely because trade policies enhanced food security and provided a cushion against the impact of international food price rises on the domestic price of staple foods. Moreover it did not discover a positive link between trade openness and overnutrition, possibly because the sheer availability of food is not enough to change consumption patterns. It did, however, find a positive correlation between such globalisation processes as increased TV ownership and internet access and overnutrition . Foreignall forms of malnutrition’. Some of the action undertaken to date to meet this global target include: a national infant and young child feeding policy that aims to increase exclusive breastfeeding for the first 6 weeks; a new poor relief programme in addition to PATH; measures to enhance the knowledge of small farmers; a new food and nutrition policy that aims to make the country more self-sufficient in food production; the formation of a National Food Industry Task Force; and a ban on certain types of sugary drinks in and around schools  and their successor the sustainable development goals (SDGs) have provided the GoJ with an impetus to address both conventionally understood malnutrition and rising rates of childhood obesity. The SDGs, for instance, include the aim to ‘eradicate  schools . Given JThe foregoing has shown that child malnutrition in Jamaica gradually declined after independence but that there remained pockets of high child malnutrition, particularly in the poorer rural areas. The overall decline in child malnutrition rates after independence was the result of various factors: an expansion of the primary health care system from the late 1960s onwards, making it easier for mothers to access child welfare and nutrition clinics; the vernacularisation of nutrition education; the adoption of pro-poor policies, ranging from food subsidies and land leasing schemes to food stamps and PATH; and loans and food aid from international aid agencies and lenders. This integration into the global economy, however, also underpinned Jamaica’s rising rates of overweight and obesity towards the end of the 20th century. The resulting exposure to global economic downturns and to international institutions that demanded fiscal austerity worsened the food security of poorer households.The approach to child malnutrition in post-independence Jamaica shifted from a medicalised approach, focussing on treatment of severe cases, to a more holistic approach that combined treatment with prevention, and which paid attention to cultural beliefs and values, involved multiple sectors, and used lay health workers. Nonetheless, it was very much what There are some signs that the GoJ realises that traditional approaches to nutrition are ineffective to address the double burden of child malnutrition. For example, recognising the contribution of corporations to the problems of childhood obesity, it is increasingly involving the private sector in initiatives to address these issues. For instance, the creation of a National Food Industry Task Force in 2017 brought together stakeholders from the food industry and various other interests groups, including the National Consumers League, the Jamaican Agricultural Society, PAHO and the University of Technology, to develop new dietary guidelines. Also the participation of various Jamaican charities and NGOs in the Healthy Caribbean Coalition’s programme to prevent childhood obesity signals a step away from a traditional deficit-led approach.Yet it is questionable how easy it is for Caribbean SIDSs, such as Jamaica to create ‘healthy publics’ to tackle the double burden of child malnutrition. First of all, these small economies, heavily reliant on external markets for both exports and imports, are extremely vulnerable to external shocks that can significantly affect the nutritional status of their populations. Take, for example, the Covid-19 pandemic which will reduce tourism earnings, the key economic driver and foreign exchange earner in Jamaica, limiting the country’s ability to import food. The pandemic will also increase unemployment so that more families will struggle to purchase food. Furthermore, children from low-income households will miss out on nutritious school meals because of school closures. And the crisis will also drive up food prices as a result of increased demand for food, changes in the supply chain, etc. .And second, Jamaica and most other Caribbean SIDS are patronage democracies; that is, democracies in which ‘citizens, especially those in lower-income groups, are integrated into politics through clientelistic relationships with their political parties’ , p. 5. F


HNF1A, and a subtype of hepatocellular adenoma (HCA) is caused by biallelic somatic HNF1A mutations; rare HCA may be related to MODY 3. This study aimed to investigate the cosegregation of HNF1A mutations with diabetes and HCA in two families.Maturity-onset diabetes of the young type 3 (MODY 3) is a consequence of heterozygous germline mutations in HNF1A germline and somatic mutations using direct sequence analysis and methylation-specific multiplex-ligation-dependent probe amplification (MS-MLPA) assay. Further, we screened eight relatives in the two independent families for diabetes, HCA and HNF1A variants. Additionally, we reviewed the literature concerning the phenotypes of MODY 3 and HCA at the background of HNF1A mutations.Two patients suffering from HCA and diabetes were screened for HNF1A identified initially using direct sequence analysis (c.686G>A in family A and c.526 + 1G>A in family B). Somatic deletion of the second allele of HNF1A was found in liver tumor tissues in both probands who were diagnosed with HCA. There are a total of ten cases of both MODY 3 and HCA phenotypes reported in the literature to date; incomplete penetrance for HCA was observed, and all the patients with HCA developed diabetes. The onset of diabetes and HCA was highly variable, the treatment of diabetes varied from diet to insulin, and the clinical expression of HCA ranged from silent to hemorrhage. Further, the severity of diabetes mellitus was not related to the occurrence of HCA.Here we reported two families  with two missense germline mutations of HNF1A mutations and highlights the importance of screening for HCA in MODY 3 families to avoid the possibility of severe complications. Further, the current study indicated that there may be a special mutational spectrum of HNF1A correlated with HCA in MODY 3 families.This study describes the association of HCA and MODY 3 at the background of  HNF1A) gene, which accounts for nearly 2–5% of insulin independent diabetes and 30–65% of MODY and is characterized by positive glycosuria, microvascular complications tendency, and sulfonylurea priority [HNF1A is mainly expressed in pancreatic beta cells, the intestine, and the liver [HNF1A lead to decreased insulin secretion, and progressive damage of beta cells and thus cause the onset of diabetes. Hepatocellular adenoma (HCA), which is known as a liver tumor characterized as being rare and benign, is widely believed to be related to oral contraception [HNF1A, and 90% of the cases are somatic mutations [HNF1A mutations and phenotypes of both MODY 3 and HCA, sometimes with the two pathologies appearing in the same individuals [HNF1A mutations.Maturity-onset diabetes of the young (MODY) is an autosomal dominantly inherited type of diabetes that results from heterozygous mutations in various transcription factors acting in the development and function of pancreatic beta cells . MODY 3 aception . RegardlHNF1A mutations, and reviewed the literature concerning phenotypes of MODY 3 and HCA.We herein report the cosegregation phenotypes of HCA and diabetes in two unrelated Chinese MODY 3 families with germline and somatic The probands from two families exhibited early-onset noninsulin-dependent diabetes as well as HCA. HCA was discovered by noninvasive radiological imaging, and confirmed by histological evaluation Fig. , which wPeripheral blood samples (4 ml) were obtained from the two MODY probands and the relatives available from the two families. Genomic DNA was extracted from whole blood using a QIAamp DNA Mini Kit  according to the manufacturer’s recommendations .Liver tumor tissue samples collected from two probands with proven HCA by surgical biopsy were frozen immediately in liquid nitrogen and stored at −80 °C. Tumor DNA was extracted using a salting-out procedure. Informed consent was obtained from all individual participants and/or parents/guardians included in the study.HNF1A were screened for mutations in the two probands by direct sequencing of PCR products as previously described [HNF1A). The direct diagnosis of the mutation identified in the probands were then offered to relatives independently of their clinical status.The ten exons, the exon–intron boundaries, and the promoter region of escribed . The NCBHNF4A, GCK, HNF1A, and HNF1B genes and is therefore specific for MODY 1, 2, 3, and 5. For the HNF4A gene, 12 probes are included, furthermore 11 for the GCK gene, 11 for the HNF1A gene, and 10 for the HNF1B gene are included. In addition, eight reference probes are included in this probe mix. The identity of the genes detected by the reference probes is available online (www.mlpa.com). The validation experiment should result in a standard deviation <0.10 for all probes over the experiment . The threshold for chromosomal abnormalities was established as follows: the lower limit was 0.7 for deletion and the upper limit was 1.33 for duplication. Abnormal values were plotted outside the threshold line. Probes for HNF1A are listed in Supplementary Table The direct sequencing of PCR products and the methylation-specific multiplex-ligation-dependent probe amplification (MS-MLPA) assay were performed in tumor DNA. MS-MLPA was performed with SALSA MLPA Probemix P241-E1 MODY Mix 1  in Beijing Novocardio Biotechnology Co., Ltd. The P241-E1 MODY Mix 1 Probemix contains 52 MLPA probes with amplification products between 130 and 500 nt. It contains probes for the HNF1A-MODY”, and liver neoplasms, such as “liver tumor”, “hepatocellular adenoma” and “liver neoplasms”, were used in various combinations and permutations across the databases. Language restriction (English) was applied. There were a total of four papers that included ten patients that met the requirements [The literature search occurred in March 2019. We systematically identified all potentially relevant articles from the following three electronic databases: MEDLINE, PubMed and Web of Science. Search terms about diabetes such as “maturity-onset diabetes of the young (MODY)” and “irements –9  in HNF1A was found in proband 2, and in other three relatives in family 2, and the mutation cosegregated with the clinical phenotypes of MODY 3 within the pedigree. The mutation is in the first nucleotide of intron 2 in HNF1A, and this variant is a splicing mutation that may affect protein function and cause diabetes. Neither of the two mutations were found in the ExAC database [The familial history of diabetes and the age at diagnosis of diabetes in both families suggested that the two probands may be diagnosed as MODY Fig. ; Table 1nds Fig. . A heterdatabase .Fig. 1Pe2. Laboratory testing was only notable for elevated fasting glucose (7.5 mmol/L), 2 h-postprandial glucose (9.1 mmol/L) and glycated albumin (17.3%) in proband 1 and for elevated 2 h-postprandial glucose (11.3 mmol/L) in proband 2.The clinical manifestations of diabetes in the two families were extraordinarily variable, the age of onset ranged from 13 to 52, and the treatment varied from diet to sulfonylurea. Further, they had BMI ranges from 19.2 to 24.3 kg/mHCA was detected in the two probands but was not in other relatives diagnosed with MODY 3. In both probands, patients did not have abdominal symptoms or liver enzyme disturbances, and therefore HCA was diagnosed through ultrasonography screening. The tumor markers  were normal in both probands. Computed tomography (CT) scans were performed in the two probands  is the most abundant miRNA in the liver, and its deregulation is associated with hepatocellular carcinoma metastasis. Interestingly, miR-122 is under the transcriptional control of HNF1A. These results further indicate the strong correlation between HNF1A and liver adenomas.HNF1A is a transcription factor that is expressed in a series of tissues, including the pancreas, liver, and digestive tract. However, the role of function , and thec nodules; thus, HHNF1A mutations are found in ~30% of HCAs; however, most of these patients do not suffer from MODY 3. The mutations are biallelic in most cases of HCA, which is consistent with the role of HNF1A as a tumor suppressor [HNF1A contain the majority of the DNA binding domain, and mutations in these positions may result in a dominant-negative effect on HNF1A function, thus leading to the high penetrance of HCA in carriers of those specific mutations. In the current study, we found biallelic mutations in the DNA binding domain of HNF1A in the tumor tissues of the two probands. However, this phenomenon was only observed in two patients suffering from both HCA and MODY 3 in the literature [HNF1A may occur outside the dimerization domain in patients with MODY 3; therefore, the encoded proteins may dimerize with the wild-type protein but disrupt transcriptional function [HNF1A may be associated with the development of HCA.ppressor . Exons 2terature . Furtherfunction . Consistfunction , 17. Thefunction . The splTo date, there are a total of four case reports describing ten patients with MODY 3 and HCA –9; the eHNF1A, and advised systematic screening of HCA through noninvasive liver imaging in MODY 3 pedigrees. In the future, additional studies may help to establish the prevalence of HCA in MODY 3 patients and the potential mechanism beneath this connection.In summary, our study enabled us to evaluate the cosegregation of HCA and MODY 3 with loss of function mutations of Supplementary Table 1


Editor-in-ChiefRob DeSalle - American Museum of Natural History, 79th Street at Central Park W, New York, NY 10024, USA. http://desalle.amnh.orgAssociate EditorMikhail Alexeyev - University of South Alabama, Dept. Physiology and Cell Biology, 5851 USA Dr. North, MSB3074, Mobile, AL 36688, USAManaging & Handling EditorSergios-Orestis Kolokotronis - SUNY Downstate Medical Center, School of Public Health, 450 Clarkson Avenue, Mail Stop Code 43, Brooklyn, NY 11203-2098, USA


Paenibacillus polymyxa are plant growth-promoting rhizobacteria (PGPR) with the potential to promote plant growth and suppress phytopathogens and have been used as biological control agents (BCAs). However, the growth promotion and biocontrol mechanisms of P. polymyxa have not been thoroughly elucidated thus far. In this investigation, the genome sequences of two P. polymyxa strains, ZF129 and ZF197, with broad anti-pathogen activities and potential for growth promotion were comparatively studied. Comparative and functional analyses of the two sequenced P. polymyxa genomes showed that the ZF129 genome consists of one 5,703,931 bp circular chromosome and two 79,020 bp and 37,602 bp plasmids, designated pAP1 and pAP2, respectively. The complete genome sequence of ZF197 consists of one 5,507,169 bp circular chromosome and one 32,065 bp plasmid, designated pAP197. Phylogenetic analysis revealed that ZF129 is highly similar to two P. polymyxa strains, HY96-2 and SQR-21, while ZF197 is highly similar to P. polymyxa strain J. The genes responsible for secondary metabolite synthesis, plant growth-promoting traits, and systemic resistance inducer production were compared between strains ZF129 and ZF197 as well as other P. polymyxa strains. The results indicated that the variation of the corresponding genes or gene clusters between strains ZF129 and ZF197 may lead to different antagonistic activities of their volatiles or cell-free supernatants against Fusarium oxysporum. This work indicates that plant growth promotion by P. polymyxa is largely mediated by phytohormone production, increased nutrient availability and biocontrol mechanisms. This study provides an in-depth understanding of the genome architecture of P. polymyxa, revealing great potential for the application of this bacterium in the fields of agriculture and horticulture as a PGPR.Many bacteria belonging to  Paenibacillus polymyxa (formerly Bacillus polymyxa), which is an important beneficial member of the PGPR, has been applied in the fields of agriculture and horticulture in the form of soil inoculants to control a wide array of plant pathogens  have been identified as environmentally friendly alternatives to traditional agrochemicals for improving crop yield and quality . Paenibaathogens . P. polyigitatum  and the nacearum . AlthougP. polymyxa has been reported to produce various potent antimicrobial and volatile compounds that reduce plant disease severity . Currently, comparative genomics is recognized as an important tool for identifying and understanding major biocontrol mechanisms and key functional genes among related organisms , and compare the genomes with the thirteen representative P. polymyxa strains that are beneficial to plant growth. Strains ZF129 and ZF197 both exhibit significant broad inhibitory spectra against various plant-pathogenic fungi and bacteria and possess excellent biocontrol characteristics and potential for the biocontrol of vegetable diseases. However, their specific biocontrol mechanisms, especially at the molecular level, are still unclear. By comparing the genomic analysis with the P. polymyxa strains, our aim was to better understand their biocontrol mechanisms at the molecular level. Besides, secondary metabolite biosynthesis, IAA biosynthesis, phosphate solubilization, nitrogen fixation, and systemic resistance inducer production were analyzed via genomic comparison. These data will provide important insights for the study of biocontrol mechanisms and benefit the practical application of strains ZF129 and ZF197 in the field.In this study, we demonstrate the sequences and annotations of two strains 600 = 0.8) using a TIANamp Bacteria DNA kit . The antagonistic activities of strains ZF129 and ZF197 against pathogenic bacteria and fungi were assessed through plate bioassays, and all of the experiments were repeated three times.Strain ZF129 and ZF197 were isolated from the rhizosphere soil of potatoes grown in Guyuan and Gaomi, respectively, in China. Strains ZF129 and ZF197 were cultivated in LB (Luria broth) medium at 28°C with shaking for 36 h. The morphology of the two strains was observed by scanning electron microscopy (SEM) and transmission electron microscopy . Genomic DNA was extracted from cultured ZF129 and ZF197 cells  = [(growth in control – growth in treatment)/growth in control] × 100.To measure the inhibitory activity against mycelial growth, the prepared cell-free supernatant was added to agar plates (1.5% w/v agar) containing potato dextrose agar  to achieve a final concentration of 10% (v/v). LB medium was used as the control. Then, a 6-mm mycelial plug was removed from the margin of the P. polymyxa ZF129 and ZF197 against F. oxysporum was performed in sealed dishes using a previously described method  of the fungal colonies were measured, and inhibition activity was calculated in the same way.A bioassay of volatiles from d method  with somP. polymyxa ZF129 and ZF197 were sequenced by Igenecode, Beijing, China1. Whole-genome sequencing was performed using the Pacific Biosciences (PacBio) RS II platform, and a 20-kb SMRTbell template was used for library construction. The sequences were assembled de novo using the HGAP v.2.3 program, which uses an overlap-layout-consensus algorithm with the parameter Genome Length of 6000000. The HGAP pipeline uses the longest reads as seeds to recruit all other reads for the construction of highly accurate preassembled reads through a directed acyclic graph-based consensus procedure, which we follow with assembly using off-the-shelf long-read assemblers (2)  database5 (The genomes of semblers . Graphicsemblers . The idelers (2) . Transfelers (2) . The funs (2)  . The gene sequences were aligned using MUSCLE and trimmed to remove ambiguously aligned regions. Subsequently, the phylogenetic tree was constructed using the maximum likelihood method in MEGA 6.0  and P. polymyxa SQR-21 (GenBank Accession No. CP006872.1), were selected for genome comparison with ZF129. Likewise, P. polymyxa J (GenBank Accession No. CP015423.1) was selected for genome comparison with ZF197. Average nucleotide identities (ANIs) (in silico DNA-DNA hybridization (DDH) 7, respectively. Furthermore, complete genome comparisons were conducted with the progressive alignment option of Mauve 2.3.1 comparison software using the ZF129 and ZF197 genomes as the reference genomes  based on six housekeeping genes   and in son (DDH)  were cal genomes . The Mau genomes .8. Comparative analyses of secondary metabolite gene clusters identified among P. polymyxa ZF129, P. polymyxa ZF197, P. polymyxa HY96-2, P. polymyxa SQR-21, P. polymyxa SC2, and P. polymyxa J were performed based on the Kyoto Encyclopedia of Genes and Genomes database (KEGG9) and the GenBank database.Secondary metabolite gene clusters prediction was performed using antiSMASH 2.0 on the authors’ Web servers using the default parametersP. polymyxa ZF129, P. polymyxa ZF197, P. polymyxa HY96-2, P. polymyxa SQR-21, P. polymyxa SC2, and P. polymyxa J using the BLAST .Functional genes involved in plant growth promotion or plant-bacterial interactions, such as those genes responsible for Indole-3-Acetic Acid (IAA) production, phosphate solubilization, and nitrogen fixation, were searched in the NCBI and KEGG databases. The identities of different functional genes at the amino acid level were compared between F. oxysporum, 27 bacteria were isolated from the rhizosphere of potato plants. Among these bacteria, two strains, ZF129 and ZF197, exhibited the highest inhibitory rates of 60.26 and 59.83%, respectively , while the inhibition rate of the cell-free supernatant of ZF129 against F. oxysporum was just 7.73%  and inhibited the mycelial growth of ectively . The celst 7.73% . After tPaenibacillus polymyxa ZF129 and ZF197 were determined to be motile, Gram-positive, rod-shaped, endospore-forming, facultative anaerobic bacteria belonging to the Paenibacillaceae family , P. polymyxa SQR-21 (46.50%), and P. polymyxa J (45.70%), but higher than those of P. polymyxa M1 (44.80%) and P. polymyxa SC2 (44.58%) (P. polymyxa ZF129 and ZF197 are presented in The complete genome of  plasmid . The ave(44.58%) . GraphicThe functional categorization of the CDSs of the ZF129 and ZF197 genomes was analyzed using the Clusters of Orthologous Groups of proteins (COG) database . The resP. polymyxa ZF129 and ZF197, five publicly available complete genome sequences of P. polymyxa strains, including HY96-2, SQR-21, SC2, M1, and J, were selected   . As expepolymyxa . Based oBacillus standard strains were used as reference genomes. These findings revealed that strains ZF129, HY96-2, and SQR-21 clustered closely with each other and occupied the same taxonomic position.Average nucleotide identity and DNA-DNA hybridization analyses are widely used to calculate whole-genome sequence similarities by comparing genomic data, and strains exhibiting ANI values ≥96% and DDH values ≥70% are typically regarded as the same species . In thisP. polymyxa strains, the whole-genome sequences were compared using Mauve. The alignments between ZF129 and ZF197 showed that several gene inversions or deletions were detectable in P. polymyxa ZF129 , three encoding transAT-PKSs (trans-acyl transferase polyketide synthetases) or NRPSs, one encoding an NRPS-like protein, three encoding lanthipeptides, and one each encoding one lassopeptide, one betalactone and one bacteriocin. Ten gene clusters related to secondary metabolite production were retrieved in strain ZF197, including five encoding NRPSs and one each encoding an NRPS-like protein, one lassopeptide, one bacteriocin, one NRPS or beatlactone and one phosphonate .The comparison of genes or gene clusters related to antibiotic synthesis suggested that five secondary metabolites could be synthesized by strains ZF129, HY96-2, SQR-21, ZF197, and J, including fusaricidin, tridecaptin, paeninodin, and two unknown antibiotics. Furthermore, 14 gene clusters related to the biosynthesis of secondary metabolites existed in strains ZF129, HY96-2, and SQR-21 with high identities , and sevP. polymyxa ZF129 and ZF197 were retrieved from the NCBI and KEGG databases, and the identities of these genes among ZF129, ZF197, HY96-2, SQR-21, and J were compared. The results indicated that six key genes  responsible for IAA (indole-3-acetic acid production) production were all found in the genomes of ZF129, ZF197, HY96-2, SQR-21 and J, with sequence identities exceeding 94% between P. polymyxa ZF129 and the other strains  were only found in the genomes of strains ZF197 and J , methanethiol , and isoprene  synthesis were found in the genomes of all these strains, with sequence identities exceeding 92% . Phylogenetic trees, which were constructed based on the 16S rRNA gene and five housekeeping genes  using maximum likelihood methods, show significant difference with each other. The phylogenetic trees show that the eleven P. polymyxa strains  form a monophyletic group. The strain ZF129 was clustered with HY96-2 and SQR-21 in Poly-clade subspecies. However, two P. polymyxa strains (J and ZF197) form the other monophyletic group. Population structure analysis also support that the Poly-clade strains evolved from a common ancestor.In this study, the evolutionary position of ZF129 and ZF197 relative to other eleven P. polymyxa genomes are presented in General features of the seven completely sequenced P. polymyxa strains from different branches of the phylogenetic tree was analyzed on the basis of ANI and DDH. ANI and DDH values among the representative P. polymyxa strains are shown in P. polymyxa genomes were no significant difference except strain J. Lower ANI similarity (<90%) was obtained when P. polymyxa J were used as reference genomes and DDH values were lower than 70%. This genetic difference maybe due to host-specific strain diversity, adaptation and ecological niche. A possible explanation for the result may that strain ZF129 and ZF197 belonged to different subspecies in P. polymyxa subspecies.Average nucleotide identity and DNA-DNA hybridization were used as effective tools for phylogenetic analysis at the genomic level. Strains with ANI values >96% and DDH values ≥70% are considered to be the same species . Genome P. polymyxa genomes  totaling 32.4 kb and one large ORF (fusA) of approximately 23.7 kb, encoding a six-module non-ribosomal peptide synthetase involved in fusaricidin production  and gad (encoding gluconic acid dehydrogenase) are the main genes responsible for the production of gluconic acid and its conversion (gcd and gad have been reported in P. polymyxa strains M1 and E681 (phn) is responsible for the bacterial degradation of phosphonates, which release biologically available phosphate for nearby plants. Many P. polymyxa strains have been reported to carry phn genes (phnABCDEWXM), including strains CR1 and E681  is one of the major nutrients in plants, second only to nitrogen in terms of its requirement, and is considered a major essential macronutrient for the growth and development of plants . The minnversion . Howeverand E681 . Anotherand E681 ; howeverB. subtilis  is an ATP-binding protein , and this operon might be associated with phosphonate uptake in response to phosphate deficiency, which needs to be further verified.In addition, the PST (phosphate-specific transport) system, a high-affinity, low-velocity, free-Pi transport system, serves as a major Pi transport system in subtilis . The pst protein . Our com2) into ammonium (NH4), thereby improving plant growth and crop yields by increasing the concentration of biologically available nitrogen  genes vary significantly among the different N2-fixing organisms, which raises the question of the origins and evolution of Monitrogenase  have been found to be related to nitrogen fixation  was identified only in P. polymyxa ZF197 and P. polymyxa J and not in three other P. polymyxa strains, ZF129, HY96-2 and SQR-21. Nevertheless, the 7 nif genes identified in ZF197 are of particular interest because of their potential use as a source of transferable genetic elements related to nitrogen fixation to facilitate the development of genetically modified PGPR.Nitrogen is an important limiting element for plant growth and production in agricultural systems since plants only absorb reduced forms of nitrogen, such as ammonia and nitrates. Nitrogenase is an oxygen-sensitive dinitrogen reductase that is produced by certain microorganisms . It can nitrogen . NitrogeChlorobi . P. polyfixation . For ins enzymes . Howeverrogenase . For exaeumoniae , wherease genome . It has fixation . The nifynthesis . Howeverdentical . Our resBacillus spp. were shown to be novel determinants of ISR elicitation in Arabidopsis  production, although their sequence identities were different. The results indicated that the ZF129, ZF197, HY96-2, SQR-21, and J strains could induce similar systemic resistance in plants but with varying efficacies owing to the differences in the related genes.Induced systemic resistance is one of the biocontrol mechanisms whereby beneficial microorganisms control plant diseases . ISR, whbidopsis . The VOCla ES426 . In thisP. polymyxa ZF129 and ZF197 have been deposited in NCBI GenBank under accession numbers CP040829.1, CP040830.1, CP040831.1, CP042272.1, and CP042273.1, respectively. The two strains have also been deposited in the China General Microbiological Culture Collection Center (CGMCC) for Type Culture Collection under accession numbers 17631 and 17632, respectively.The complete genome sequences of QW and J-YL conceived and designed the experiments. J-YL performed the experiments. J-YL, T-TG, and QW analyzed the data and wrote the manuscript. All authors have read and approved the final manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.