Floral morph variation mediated by clonal growth and pollinator functional groups of Limonium otolepis in a heterostylous fragmented population.

Abstract. Heterostyly, a genetic style polymorphism, is linked to symmetric pollen transfer, vital for its maintenance. Clonal growth typically impacts sexual reproduction by influencing pollen transfer. However, the floral morph variation remains poorly understood under the combined effects of pollinators and clonal growth in heterostyly characterized by negative frequency-dependent selection and disassortative mating. We estimated morph ratios, ramets per genet and heterostylous syndrome and quantified legitimate pollen transfer via clonal growth, pollinators and reciprocal herkogamy between floral morphs in Limonium otolepis, a fragmented population composed of five subpopulations in the desert environment of northwestern China, with small flower and large floral morph variation. All subpopulations but one exhibited pollen-stigma morphology dimorphism. The compatibility between mating types with different pollen-stigma morphologies remained consistent regardless of reciprocal herkogamy. Biased ratios and ramets per genet of the two mating types with distinct pollen-stigma morphologies caused asymmetric pollen flow and varying fruit sets in all subpopulations. Short-tongued insects were the primary pollinators due to small flower sizes. However, pollen-feeding Syrphidae sp. triggered asymmetry in pollen flow between high and low sex organs, with short-styled morphs having lower stigma pollen depositions and greater variation. Clonal growth amplified this variation by reducing intermorph pollen transfer. All in all, pollinators and clonal growth jointly drive floral morph variation. H-morphs with the same stigma-anther position and self-incompatibility, which mitigate the disadvantages of sunken low sex organs with differing from the classical homostyly, might arise from long- and short-styled morphs through a 'relaxed selection'. This study is the first to uncover the occurrence of the H-morph and its associated influencing factors in a distylous plant featuring clonal growth, small flowers and a fragmented population.

Asymmetrical disassortative pollination mediated by long-/short-tongued pollinators in a distylous Limonium myrianthum (Plumbaginaceae) with a short corolla tubular small flower.

In heterostylous plants, short-tongued pollinators are often ineffective/inefficient owing to the limitations imposed by a long corolla tube. However, it is unclear how disassortative pollen transfer is achieved in small flowers. We investigated the pollination pattern and floral morph variation by analyzing heterostylous syndrome, pollinator groups, and pollen deposition after a single visitation in two Limonium myrianthum populations with short-corolla-tubular small flowers. The predominant pollinators in the Hutubi population were pollen-seeking short-tongued syrphids, which can only transfer pollen between high-level sexual organs. In the Xishan population, nectar-seeking short-tongued insects were efficient pollinators with symmetrical disassortative pollen transfer between high- and low-level sexual organs, whereas long-tongued pollinators had a low efficiency between high-level sexual organs due to the low contact probability with the stigma of long-styled flowers (L-morph), which no longer offered the same advantage observed in tubular flowers. Asymmetrical disassortative pollination may cause the female fitness of short-styled (S-morph) individuals in the Hutubi and L-morph individuals in the Xishan population to suffer greater selection pressure and exhibit a higher degree of floral morph variation. Limonium myrianthum exhibits an unusual pollination pattern in which the small flowers with short corolla tubes make it possible for short-tongued insects to become effective pollinators. However, factors such as the position of stigma-anther within the flower, pollinator species and their preference further caused asymmetrical disassortative pollen transfer. Therefore, more factors should be considered when evaluating the effectiveness of short- and long-tongued insects in pollination service.

Synergistic inhibition mechanism of quinazolinone and piperacillin on penicillin-binding protein 2a: a promising approach for combating methicillin-resistant Staphylococcus aureus.

The production of penicillin-binding protein 2a (PBP2a), a cell wall synthesis protein, is primarily responsible for the high-level resistance observed in methicillin-resistant Staphylococcus aureus (MRSA). PBP2a exhibits a significantly reduced affinity for most ß-lactam antibiotics owing to its tightly closed active site. Quinazolinones (QNE), a novel class of non-ß-lactam antibiotics, could initiate the allosteric regulation of PBP2a, resulting in the opening of the initially closed active pocket. Based on our previous study, we have a basic understanding of the dual-site inhibitor ceftaroline (CFT) induced allosteric regulation of PBP2a. However, there are still limitations in the knowledge of how combining medicines, QNE and piperacillin (PIP), induce the allosteric response of PBP2a and inhibit its function. Herein, molecular dynamics (MD) simulations were performed to elucidate the intricate mechanisms underlying the combination mode of QNE and PIP. Our study successfully captured the opening process of the active pocket upon the binding of the QNE at the allosteric site, which alters the signaling pathways with a favorable transmission to the active site. Subsequent docking experiments with different conformational states of the active pocket indicated that all three inhibitors, PIP, QNE, and CFT, exhibited higher docking scores and more favorable docking poses to the open active pocket. These findings reveal the implied mechanism of QNE-mediated allostery underlying combination therapy and provide novel insights into developing innovative therapeutic modalities against MRSA.Communicated by Ramaswamy H. Sarma.

Unraveling the atomic mechanisms underlying glyphosate insensitivity in EPSPS: implications of distal mutations.

5-enolpyruvyl shikimate-3-phosphate synthase (EPSPS), as an indispensable enzyme in the shikimate pathway, is the specific target of grasser killer glyphosate (GPJ). GPJ is a competitive inhibitor of phosphoenolpyruvate (PEP), which is the natural substrate of EPSPS. A novel Ls-EPSPS gene variant discovered from Liliaceae, named ELs-EPSPS, includes five distal mutations, E112V, D142N, T351S, D425G, and R496G, endowing high GPJ insensitivity. However, the implicit molecular mechanism of the enhanced tolerance/insensitivity of GPJ in ELs-EPSPS is not fully understood. Herein, we try to interpret the hidden molecular mechanism using computational methods. Computational results reveal the enhanced flexibility of apo EPSPS upon mutations. The enhanced affinity of the initial binding substrate shikimate-3-phosphate (S3P), and the higher probability of second ligands PEP/GPJ entering the pocket are observed in the ELs-EPSPS-S3P system. Docking and MD results further confirmed the decreased GPJ-induced EPSPS inhibition upon mutations. And, the alterations of K98 and R179 side-chain orientations upon mutations are detrimental to GPJ binding at the active site. Additionally, the oscillation of side chain K98, in charge of PEP location, improves the proximity effect for substrates in the dual-substrate systems upon mutations. Our results clarify that the enhanced GPJ tolerance of EPSPS is achieved from decreased competitive inhibition of GPJ at the atomic perspective, and this finding further contributes to the cultivation of EPSPS genes with higher GPJ tolerance/insensitivity and a mighty renovation for developing glyphosate-resistant crops.Communicated by Ramaswamy H. Sarma.

Effects of adding milk to fermented black mulberry (Morus nigra L.) juice on its antioxidant activity in C2C12 cells and changes in volatile flavor compounds during storage.

This study assessed the impact of milk on the bioactive compounds, physicochemical properties, antioxidant activity, ROS inhibition, and volatile flavor compounds of fermented black mulberry juice (FBMJ). Firstly, the results showed that 25% concentration of milk was the most suitable for preparing FBMJ-Milk. Compared to the control group, the addition of milk significantly increased the SOD activity and antioxidant capacity, as well as enhanced the total phenolic content (TPC) and SOD storage stability. Secondly, HS-SPME-GC-MS combined with OPLS-DA analysis identified 49 compounds in FBMJM, including 12 esters, 6 acids, 1 ketone, 2 aldehydes, 19 alcohols and 9 other compounds. During the storage, the levels of ethyl ester compounds decreased significantly, while the degradation of ester produced some acid and alcohol compounds. The findings revealed that the addition of milk was beneficial for maintaining the antioxidant stability of FBMJM during storage and enhancing the richness of product flavor.

Unraveling the mechanism of ceftaroline-induced allosteric regulation in penicillin-binding protein 2a: insights for novel antibiotic development against methicillin-resistant Staphylococcus aureus.

Methicillin-resistant Staphylococcus aureus (MRSA) acquires high-level resistance against ß-lactam antibiotics by expressing penicillin-binding protein 2a (PBP2a). PBP2a is a cell wall-synthesizing protein whose closed active site exhibits a reduced binding affinity toward ß-lactam antibiotics. Ceftaroline (CFT), a fifth-generation cephalosporin, can effectively inhibit the PBP2a activity by binding to an allosteric site to trigger the active site opening, allowing a second CFT to access the active site. However, the essential mechanism behind the allosteric behavior of PBP2a remains unclear. Herein, computational simulations are employed to elucidate how CFT allosterically regulates the conformation and dynamics of the active site of PBP2a. While CFT stabilizes the allosteric domain surrounding it, it simultaneously enhances the dynamics of the catalytic domain. Specifically, the study successfully captured the opening process of the active pocket in the allosteric CFT-bound systems and discovered that CFT alters the potential signal-propagating pathways from the allosteric site to the active site. These findings reveal the implied mechanism of the CFT-mediated allostery in PBP2a and provide new insights into dual-site drug design or combination therapy against MRSA targeting PBP2a.

Molecular Dynamics Simulation of Transport Mechanism of Graphene Quantum Dots through Different Cell Membranes.

Exploring the mechanisms underlying the permeation of graphene quantum dots (GQDs) through different cell membranes is key for the practical application of GQDs in medicine. Here, the permeation process of GQDs through different lipid membranes was evaluated using molecular dynamics (MD) simulations. Our results showed that GQDs can easily permeate into 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) and 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) lipid membranes with low phospholipid molecule densities but cannot permeate into 1-palmitoyl-2-oleoyl phosphatidylethanolamine (POPE) lipid membranes with high phospholipid densities. Free energy calculation showed that a high-energy barrier exists on the surface of the POPE lipid membrane, which prevents GQDs from entering the cell membrane interior. Further analysis of the POPE membrane structure showed that sparsely arranged phospholipid molecules of the low-density lipid membrane facilitated the entry of GQDs into the interior of the membrane, compared to compactly arranged molecules in the high-density lipid membrane. Our simulation study provides new insights into the transmembrane transport of GQDs.

Theoretical Investigation of Switch Effect on the Efficiency and Adaptivity of Molecular Optoelectronic Conversion Devices.

Molecular photoswitch can effectively regulate charge separation (CS) and charge recombination (CR) in donor-acceptor (D-A) systems. However, deformation of the donor-switch-acceptor (D-S-A) systems caused by the switch isomerization will destroy the geometrical stability of the battery. Here we take the planar platinum(II) terpyridyl complex of [Pt(t Bu3 tpy)(-C≡C-Ph)n ]+ as the typical D-A model, designed six D-S-A systems using different photoswitches (dimethyldihydropyrene, fulgimide, arylazopyrazole, N-salicylideneaniline, spiropyran, and dithienylethene, denoted as D-S-A 1-6 hereafter). Our investigations show that the D-S-A 1-6 can absorb visible light of 799 nm, 673 nm, 527 nm, 568 nm, 616 nm, and 629 nm, facilitating electrons transfer from the donor and the switch to the acceptor through the Switch-on channel. Then cationic character of the photoswitch can undergo much more rapid isomerization than the neutral form due to the lower energy barrier. The Switch-off isomer breaks the conjugation of the D-S-A system, effectively turning off the CT channel and forming the CS state. Based on the evaluated conjugated backbone twist (CBT) angle, we found that D-S-A 1, 2, 4, 6 exhibit little configurational change and can be good candidates as the organic solar cell. The proposed D-S-A design controlled by the molecular switch may help to develop a solution for solar-harvesting practical applications.

Designing Self-Adaptive Donor-Switch-Acceptor Systems for Molecular Opto-Electronic Conversion Based on Dimethyldihydropyrene/Cyclophanediene.

The introduction of a self-adaptive molecular switch is an appealing strategy to achieve complete charge separation (CS) in donor-acceptor (D-A) systems. Here, we designed donor-switch-acceptor (D-S-A) systems using a platinum(II) terpyridyl complex as the acceptor, dimethyldihydropyrene/cyclophanediene (DHP/CPD) as the bridge, and methoxybenzene, thieno[3,2-b]thiophene, 2,2'-bifuran, and 4,8-dimethoxybenzo[1,2-b:4,5-b']difuran as donors, respectively. We then systematically studied the whole opto-electronic conversion process of the donor-DHP/CPD-acceptor (D-DHP/CPD-A) systems based on time-dependent density functional theory, time-dependent ultrafast electron evolution, and electron transport property calculations. We first found that the substitution of -CH3 by -H and -CN groups in DHP/CPD can enlarge the range of the adsorption wavenumber in opto-electric conversion. Then the light absorption induces the cationization of DHP switch, largely accelerating the forth-isomerization to CPD form. Once the D-CPD-A molecule is formed, the poor conjugation can realize the complete CS state by inhibiting the radiative and nonradiative charge recombinations. Finally, the repeatable and complete CS can be achieved through the automatic back-isomerization of CPD to DHP. The present work provides valuable insights into design of D-S-A systems for practical utilization of molecule-based solar harvesting.

Functional vulnerability of liver macrophages to capsules defines virulence of blood-borne bacteria.

Many encapsulated bacteria use capsules to cause invasive diseases. However, it remains largely unknown how the capsules enhance bacterial virulence under in vivo infection conditions. Here we show that the capsules primarily target the liver to enhance bacterial survival at the onset of blood-borne infections. In a mouse sepsis model, the capsules enabled human pathogens Streptococcus pneumoniae and Escherichia coli to circumvent the recognition of liver-resident macrophage Kupffer cells (KCs) in a capsular serotype-dependent manner. In contrast to effective capture of acapsular bacteria by KCs, the encapsulated bacteria are partially (low-virulence types) or completely (high-virulence types) "untouchable" for KCs. We finally identified the asialoglycoprotein receptor (ASGR) as the first known capsule receptor on KCs to recognize the low-virulence serotype-7F and -14 pneumococcal capsules. Our data identify the molecular interplay between the capsules and KCs as a master controller of the fate and virulence of encapsulated bacteria, and suggest that the interplay is targetable for therapeutic control of septic infections.

Study on the Efficacy and Safety of Ambroxol Combined with Methylprednisolone in Patients with Acute Lung Injury.

BACKGROUND: There is no better treatment method towards paraquat-induced acute lung injury (ALI) at present. Ambroxol combined with methylprednisolone exhibits a significant improvement effect on ALI treatment, whereas their mechanism in ALI is still unclear. METHODS: 64 patients with ALI caused by paraquat poisoning brought to our hospital from January 2015 to January 2018 were selected. They were separated into a combined treatment group (CTG) and a routine treatment group (RTG) on the basis of different treatment methods. The survival of patients was observed after 7 days of treatment. Arterial blood gas, oxygen partial pressure (PaO2), partial pressure of carbon dioxide (PaCO2), oxygenation index (PaO2/FiO2), patient's spontaneous respiratory rate (RR), tidal volume (VT), and positive end-expiratory pressure (PEEP) were observed before and after treatment for 7 days. Interleukin 6 (IL-6) and tumor necrosis factor α (TNF-α) were analyzed. The differences of indexes between the dead patients and the survivors were observed, and the potential predictive value of death was analyzed. RESULTS: After treatment, the indexes of patients were significantly improved in both groups compared with those before therapy. Further comparison showed that the improvement of PaO2, PaCO2, and PaO2/FiO2 in CTG was obviously higher than that in RTG (p < 0.05). The improvement of RR, PEEP, and VT in CTG was obviously higher than that in RTG (p < 0.05). The decreased degree of IL-6 and TNF-α in CTG was higher than that in RTG (p < 0.05). The 7-day mortality rate of 64 patients was 39.06%, and there was no obvious difference in the 7-day survival rate in both groups (p = 0.649). IL-6 and TNF-α were expected to be potential prediction indexes of paraquat-induced ALI. CONCLUSION: Ambroxol combined with methylprednisolone significantly improved the oxygen partial pressure and oxygenation index of patients with paraquat-induced ALI and inhibited the inflammatory response of patients.

Corrigendum: Identification of Novel Laminin- and Fibronectin-Binding Proteins by Far-Western Blot: Capturing the Adhesins of Streptococcus suis Type 2.

[This corrects the article DOI: 10.3389/fcimb.2015.00082.].

Theoretical Evaluation of DNA Genotoxicity of Graphene Quantum Dots: A Combination of Density Functional Theory and Molecular Dynamics Simulations.

Owing to their unique morphology, ultrasmall lateral sizes, and exceptional properties, graphene quantum dots (GQDs) hold great potential in many applications, especially in the fields of electrochemical biosensors, bioimaging, drug delivery, gene delivery, etc. Their biosafety and potential genotoxicity to human and animal cells have been a growing concern in recent years. Especially, the potential DNA damage caused by GQDs is very crucial but still unclear. In this study, the effect of GQDs on DNA damage has been evaluated by a combination of molecular dynamics (MD) simulations and density functional theory. Our results demonstrate that the DNA damaging mechanism of GQDs depends on the size of GQDs. The small GQDs (seven benzene rings) tend to enter into the interior of DNA molecules and cause a DNA base mismatch. The relatively large GQDs (61 benzene rings) tend to adsorb onto the two ends of a DNA molecule and cause DNA unwinding. Due to the strong interaction between guanine (G) and GQDs, the effect of GQDs is much larger on G than on the other three bases (A, C, and T). In addition, the concentration of GQDs could also affect the results of DNA damaging.

RACK7 recognizes H3.3G34R mutation to suppress expression of MHC class II complex components and their delivery pathway in pediatric glioblastoma.

Histone H3 point mutations have been identified in incurable pediatric brain cancers, but the mechanisms through which these mutations drive tumorigenesis are incompletely understood. Here, we provide evidence that RACK7 (ZMYND8) recognizes the histone H3.3 patient mutation (H3.3G34R) in vitro and in vivo. We show that RACK7 binding to H3.3G34R suppresses transcription of CIITA, which is the master regulator of MHC (major histocompatibility complex) class II molecules and genes involved in vesicular transport of MHC class II molecules to the cell surface, resulting in suppression of MHC class II molecule expression and transport. CRISPR-based knock-in correction of the H3.3G34R mutation in human pediatric glioblastoma (pGBM) cells significantly reduces overall RACK7 chromatin binding and derepresses the same set of genes as does knocking out RACK7 in the H3.3G34R pGBM cells. By demonstrating that H3.3G34R and RACK7 work together, our findings suggest a potential molecular mechanism by which H3.3G34R promotes cancer.

[Effect of sonication on results of ChIP-seq experiments involving PRC2 related proteins].

Sonication is one of the essential strategies of chromatin fragmentation in Chromatin immunoprecipitation (ChIP) assay. The impact of proteins with different molecular weights generated under different duration of sonication on the results of Chromatin immunoprecipitation sequencing (ChIP-seq) experiments involving the Polycomb Repressive Complex 2 (PRC2) related protein EZH2, which is a histone methyltransferase, and its product H3K27me3 was investigated. The results indicate that in the promoter region or nonpromoter region, there were hardly any differences among the H3K27me3 peaks annotated genes from different duration of sonication, which suggesting that the duration of sonication had little effect on histone ChIP-seq results. In contrast, in the promoter region newly gained EZH2 peaks annonated genes at 20 min sonication time were significantly clustered in the gene ontology (GO) pathways related to actin filament bundle compared with 10 min. In the nonpromoter region, compared with 10 min, the GO pathways of newly gained EZH2 peaks annonated genes at 20 min sonication time is much more than that of lost EZH2 peaks annonated genes. And in the nonpromoter region, compared with 20 min, the GO pathways of lost EZH2 peaks annonated genes at 30 min sonication time is much more than that of newly gained EZH2 peaks annonated genes. Most of these pathways are associated with RNA polymerase II (RNAPII), organ development and cell morphogenesis. These suggest that the genomic information of EZH2 will be lost if the duration of sonication is not enough or too long. Different duration of sonication mainly affect the EZH2 peaks in PRC2 unoccupied region and the bivalent promoter in the promoter region, as well as the PRC2 occupied region, PRC2 unoccupied region and the activated enhancer in the nonpromoter region. Therefore, the sonication for the chromatin related proteins with high molecular weights need to be optimized to make chromatin fragments size vary from 100 bp to 500 bp, which will yield relatively comprehensive genomic information of the target protein. For histones, which are of small molecular weights, duration of sonication has little effect on them.

[Analysis of pressure-time and flow-time curve in ventilator].

OBJECTIVE: To better understand the significance of the pressure-time curve and flow-time curve from the perspective of PB840 ventilator working principle. METHODS: (1) Mechanical principle: flow supply valves (air valve and oxygen valve) and exhalation valve in PB840 ventilator were controlled to achieve the ventilation target (volume or pressure) by the central processing unit according to the monitoring data from pressure sensors (P1 at the supply side, P2 at the exhalation side) and flow sensors (Q1 at the air side, Q2 at the oxygen side, Q3 at the exhalation side). (2) The essence of curve: each point means a value of pressure or flow at a certain time measured by the sensors or calculated by the system. (3) The respiratory process could be divided into inspiratory part, expiratory part, and the connection part from expiratory to inspiratory. The air running state and the respiratory mechanics relationship at the three parts could be inferred according to the form of curves. RESULTS: (1) Inspiratory process: at volume-controlled and constant flow ventilation: there should be a relationship "Pc-Pa = XR" between alveolar pressure (Pa) and circuit pressure (Pc) according to Ohm law. So, the Pc curve (pressure-time curve) could indirectly reflect the Pa curve with the flow (X) and resistance (R) being constant. At pressure-set ventilation: it is the goal of ventilator to maintain the Pc at the target level. So, the stability of the target pressure line in pressure-time curve reflects the matching ability of the flow supply valves and the exhalation valve. (2) Expiratory process: it could be divided into pre-expiratory [without basic flow (Ba) or bias flow (Bi)] and post-expiratory (with Ba or Bi), where Ba or Bi is equal to "Q1+Q2". So, the mathematical function are "X(t) = Q3t" in pre-part, and "X(t) = Q3t-(Q1t+Q2t)" in post-part. The relationship between pressure and flow at peak expiratory flow point: it could be found that there is an obvious time span and area formation under the curve from 0 to peak point (Fpeak) after stretching the abscissa axis of flow-time curve. It means that some gas have been discharged from the lung when it arrives at the peak point. So, the alveolar pressure should be lower than the platform pressure at the point (Pplat). The circuit pressure is significantly higher than positive end expiratory pressure (PEEP) at the point in the stretching axis diagram. So, it means that the formula "RE = (Pplat-PEEP)/Fpeak" to calculate the expiratory resistance (E) is unreasonable in the angle of Ohm law. (3) The process from exhalation to inspiratory: according to the difference of the starting point of the conversion, it could be divided into two cases: one is that the inspiratory started from the ending of exhalation. Here, the inhaling starting point is lying in the abscissa axis. The other is that the inspiratory started before the ending of exhalation (with endogenous positive end expiratory pressure). Here, the starting point is lying below the abscissa axis, and the slope of the following curve is obviously larger than the slope of natural expiratory curve. According to the difference of results from the starting point to the end of the inhalation triggering effort, it could be divided into two cases: one is that it reach the trigger point. Here, the expiratory curve extends upward from or below the horizontal axis until an effective air supply is triggered. The other is that it could not reach the trigger point. Here, the expiratory curve extends upward from or below the horizontal axis, but then runs downward (meaning exhaling). CONCLUSIONS: It is helpful to analyze the ventilation state, ventilation failure, and the causes of man-machine confrontation with understanding the ventilation principle and the air route map of the ventilator.

Cost-effectiveness of a primary care multidisciplinary Risk Assessment and Management Program for patients with diabetes mellitus (RAMP-DM) over lifetime.

PURPOSE: The multidisciplinary Risk Assessment and Management Program for patients with diabetes mellitus (RAMP-DM) was found to be cost-saving in comparison with usual primary care over 5 years' follow-up. This study aimed to estimate the cost-effectiveness of RAMP-DM over lifetime. METHODS: We built a Discrete Event Simulation model to evaluate the cost-effectiveness of RAMP-DM over lifespan from public health service provider's perspective. Transition probabilities among disease states were extrapolated from a cohort of 17,140 propensity score matched participants in RAMP-DM and those under usual primary care over 5-year's follow-up. The mortality of patients with specific DM-related complications was estimated from a cohort of 206,238 patients with diabetes. Health preference and direct medical costs of DM patients referred to our previous studies among Chinese DM patients. RESULTS: RAMP-DM individuals gained 0.745 QALYs and cost US$1404 less than those under usual care. The probabilistic sensitivity analysis found that RAMP-DM had 86.0% chance of being cost-saving compared to usual care under the assumptions and estimates used in the model. The probability of RAMP-DM being cost-effective compared to usual care would be over 99%, when the willingness to pay threshold is HK$20,000 (US$ 2564) or higher. CONCLUSION: RAMP-DM added to usual primary care was cost-saving in managing people with diabetes over lifetime. These findings support the integration of RAMP-DM as part of routine primary care for all patients with diabetes.

Zc3h13 Regulates Nuclear RNA m6A Methylation and Mouse Embryonic Stem Cell Self-Renewal.

N6-methyladenosine (m6A) is an abundant modification in eukaryotic mRNA, regulating mRNA dynamics by influencing mRNA stability, splicing, export, and translation. However, the precise m6A regulating machinery still remains incompletely understood. Here we demonstrate that ZC3H13, a zinc-finger protein, plays an important role in modulating RNA m6A methylation in the nucleus. We show that knockdown of Zc3h13 in mouse embryonic stem cell significantly decreases global m6A level on mRNA. Upon Zc3h13 knockdown, a great majority of WTAP, Virilizer, and Hakai translocate to the cytoplasm, suggesting that Zc3h13 is required for nuclear localization of the Zc3h13-WTAP-Virilizer-Hakai complex, which is important for RNA m6A methylation. Finally, Zc3h13 depletion, as does WTAP, Virilizer, or Hakai, impairs self-renewal and triggers mESC differentiation. Taken together, our findings demonstrate that Zc3h13 plays a critical role in anchoring WTAP, Virilizer, and Hakai in the nucleus to facilitate m6A methylation and to regulate mESC self-renewal.

Direct medical costs of diabetes mellitus in the year of mortality and year preceding the year of mortality.

AIM: To report the health resource use and estimate the direct medical costs among patients with diabetes mellitus (DM) in the year of mortality and the year preceding the year of mortality. MATERIALS AND METHODS: We analysed data from a population-based, retrospective cohort study including all adults with a DM diagnosis in Hong Kong between 2009 and 2013, and who died between January 1, 2010 and December 31, 2013. The annual direct medical costs in the year of mortality and the year preceding the year of mortality were determined by summing the costs of health services utilized within the respective year. The costs were analysed by gender, the presence of comorbidities, diabetic complications and primary cause of death. RESULTS: A total of 10 649 patients met the eligibility criteria for analysis. On average, the direct medical costs in the year of death were 1.947 times higher than those in the year before death. Men and women with DM incurred similar costs in the year preceding the year of mortality and in the mortality year. Patients with any diabetic complications incurred greater costs in the year of mortality and the year before mortality than those without. CONCLUSIONS: This analysis provides new evidence on incorporating additional direct medical costs in the mortality year, and refining the structure of total cost estimates for use in costing and cost-effectiveness analyses of interventions for DM.

Health-related quality of life and health preference of Chinese patients with diabetes mellitus managed in primary care and secondary care setting: decrements associated with individual complication and number of complications.

BACKGROUND: Health-related quality of life (HRQoL) and health preference of patients with diabetes mellitus (DM) are essential in health economic evaluations but data on Chinese population is rare. This study aims to evaluate HRQoL and health preference of diabetic patients with different diabetic complications in Chinese population. METHODS: A cross-sectional study was conducted in 1275 patients with DM, including 518 subjects with various DM-related complications. HRQoL and health preference were estimated using SF-12 and SF-6D questionnaires, respectively. Disease status of DM and complications were identified from documented clinical diagnosis. Multivariable regression was used to investigate the effects of specific complications on HRQoL and health preference, adjusting for socio-demographic and clinical parameters. RESULTS: The presence of any diabetic complication was associated with lower physical component summary (-3.81 points, P < 0.01), and end-stage renal disease (ESRD) showed greatest reduction (-7.05 points, P < 0.01). Mental component summary and mental health (MH) scores were not decreased in any of the diabetic complications. The health preference score for diabetic subjects without complications was 0.882 (95% CI, 0.778 to 0.989). The reductions of health preference score were significant for stroke (-0.042, 95% CI -0.072 to -0.012), ESRD (-0.055, 95% CI -0.093 to -0.017), and sight-threatening diabetic retinopathy (STDR) (-0.043, 95% CI -0.075 to -0.010), while heart disease had an insignificant reduction (-0.017, 95% CI -0.042 to 0.008). CONCLUSIONS: The presence of any of the four major diabetic complications (heart disease, stroke, ESRD and STDR) was associated with lower HRQoL and health preference scores. Findings of this study facilitated the cost-effectiveness studies of alternative management strategies for prevention of diabetic complications in Chinese population.