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Recent studies have indicated the antitumor activity and reduced allogeneic response of universal chimeric antigen receptor-modified T (UCAR T) cells lacking endogenous T cell receptors and beta-2 microglobulin (B2M) generated using gene-editing technologies. However, these cells are vulnerable to lysis by allogeneic natural killer (NK) cells due to their lack of human leukocyte antigen (HLA) class I molecule expression. Here, constitutive expression of mutant B2M-HLA-E (mBE) and B2M-HLA-G (mBG) fusion proteins in anti-CD19 UCAR T (UCAR T-19) cells was conducted to protect against allogeneic NK cell-mediated lysis. The ability of cells expressing mBE or mBG to resist NK cell-mediated lysis was observed in gene-edited Jurkat CAR19 cells. UCAR T-19 cells constitutively expressing the mBE and mBG fusion proteins were manufactured and showed effective and specific anti-tumor activity. Constitutive expression of the mBE and mBG fusion proteins in UCAR T-19 cells prevented allogeneic NK cell-mediated lysis. In addition, these cells were not recognizable by allogeneic T cells. Additional experiments, including those in animal models and clinical trials, are required to evaluate the safety and efficacy of UCAR T-19 cells that constitutively express mBE and mBG.

Increasing the remission rate and reducing the recurrence rate can improve the clinical efficacy of chimeric antigen receptor (CAR) T cell therapy in recurrent/refractory non-Hodgkin lymphoma (r/rNHL). In this open-label, single-arm phase I/II trial, 87 patients with r/rNHL, including 58 patients with aggressive diffuse large B-cell lymphoma and 24 with high tumour burden, received an infusion at doses of 0.5 × 106–8 × 106 TanCAR7 T cells per kilogram of body weight after conditioning chemotherapy. The best overall response rate was 78% (95% confidence interval [CI], 68–86); response rates were consistent across prognostic subgroups. The median follow-up was 27.7 months. The median progression-free survival was 27.6 months (95% CI, 11 to not reached). Cytokine release syndrome (CRS) occurred in 61 patients (70%) with 60% of cases being grade 1 or 2 and 10% being grade 3 or greater. Grade 3 CAR T cell-related encephalopathy syndrome (CRES) occurred in 2 patients (2%). Two patients died from treatment-associated severe pulmonary infection, and one died from CRS-related pulmonary injury between 1 and 3 months post infusion. Long-term remissions were observed following the use of TanCAR7 T cells in r/rNHL with a safety profile that included CRS but few cases of CRES.

The current clinical outcome for patients with metastatic pancreatic carcinoma (PC) remains poor. Epidermal growth factor receptor (EGFR) is detectable in PC, suggesting that EGFR is a rational target in PC. We conducted a phase I clinical trial to evaluate the safety and efficacy of autologous anti-EGFR chimeric antigen receptor–modified T (CAR T–EGFR) cells in patients with metastatic PC. The expression levels of EGFR on tumor cells detected by immunohistochemistry were required to be more than 50%. Sixteen patients were enrolled and received one to three cycles of the CAR T–EGFR cell infusion within 6 months (median dose of CAR T cells: 3.48 × 106/kg; range, 1.31 to 8.9 × 106/kg) after the conditioning regimen with 100 to 200 mg/m2 nab-paclitaxel and 15 to 35 mg/kg cyclophosphamide. Grade ≥3 adverse events included fever/fatigue, nausea/vomiting, mucosal/cutaneous toxicities, pleural effusion and pulmonary interstitial exudation and were reversible. Of 14 evaluable patients, four achieved partial response for 2–4 months, and eight had stable disease for 2–4 months. The median progression-free survival was 3 months (range, 4–months) from the first cycle of CAR T–EGFR cell treatment, and the median overall survival of all 14 evaluable patients was 4.9 months (range, 2.9–30 months). Decreased EGFR expression on tumor cells was observed in patients who achieved stable disease with shrinkage of metastatic lesions in the liver, and enrichment of central memory T cells in infused cells improved the clinical response. In conclusion, the treatment with CAR T–EGFR cells is safe and effective in patients with metastatic PC. This trial was registered at www.clinicaltrials.gov (identifier no: NCT01869166).

Habitat destruction caused by human activities is threatening wetland plants. Based on Landsat 5 TM, Landsat 8 OLI, and Google Earth images, we analysed the spatiotemporal dynamics of plant populations in a degraded wetland in the Minjiang River Estuary from 2009 to 2019. Field survey records, tasselled cap transformation, and supervised classification were used for the image interpretation. The results indicate that the plant habitat area decreased by 42.1% over these 10 years. Consequently, the distribution area of each dominant species declined, and the proportion of total vegetated area in the habitat decreased from 53.1% to 32.6%. Moreover, the dominance relations of the species changed from Spartina alterniflora > Phragmites australis > Cyperus malaccensis > Scirpus triqueter to P. australis > S. alterniflora > C. malaccensis > S. triqueter. We predict that the vegetation landscape in the wetland will be composed exclusively of P. australis and S. alterniflora if habitat destruction continues. Finally, our research shows that the NPCS (net plant carbon sequestration) and PCS (plant carbon stock) in the wetland and the average values of methane flux, NPCS, PCS, and SOCS (sediment organic carbon stock) in the vegetated area have all decreased due to plant population declines.

Fresh shiitake (Lentinus edodes) is prone to brown, pileus-opening and flavor-loss during storage. Therefore, it is important to find an effective preservation method for fresh shiitake. Negative air ions (NAI) are negatively-charged molecules or atoms in the air, and can affect the physiological metabolism of live cells and be conveniently used with low cost. In this study, NAI treatment was performed at different times and the physico-chemical characteristics, microstructure, membrane potential and energy metabolism of shiitake were determined during storage. Results showed that NAI treatment for 40 min could reduce 29% of browning index and maintain the hardness of shiitake. NAI treatment groups had higher content of sweetness amino acids, umami amino acids, 5′-IMP, eight-carbon alcohols compounds and cyclic sulfides compounds than the control, and comprehensive quality of the group being treated for 40 min was the best. The mitochondria of shiitake swelled and the membrane potential decreased after being treated by NAI. However, NAI treatment for 40 min could improve the contents of ATP and ADP, maintain a relatively stable energy charge level, and promote energy utilization of shiitake during storage. The results demonstrated that NAI treatment had the potential to improve the quality shiitake during storage.

CD19-directed chimeric antigen receptor T cell (CART19) therapy is efficient and approved for relapsed/refractory diffuse large B cell lymphoma (DLBCL). To increase durable antitumor response, we previously designed tandem CART19/20 cells and shown longer progression-free survival. However, a proportion of CART19/20-treated patients will finally progress and require salvage therapies. In this study, we analyzed data from five patients with relapsed/refractory DLBCL who had disease progression or relapse following CART19/20 therapy and then treated with PD-1-blocking antibodies as salvage therapy. Two of five patients acquired complete remissions after anti-PD-1 therapy, including one patient remained ongoing remission for more than 21 months. One patient achieved a partial remission, and the other two had progressive diseases. No ≥ grade 3 treatment-related adverse events or cytokine release syndrome was observed. Immunohistochemistry of tumor specimens revealed higher PD-1/PD-L1 expression in responsive patients with anti-PD-1 therapy as compared to that in non-responders. After anti-PD-1 treatment, circulating T cells were activated in responders, and no significant expansion of CART19/20 cells was detected. Our data suggest that PD-1 blockade therapy can be active in patients with relapsed/refractory DLBCL after failure of CAR T cell therapy who had PD-L1 expression in tumor cells and high PD-1 level in tumor-infiltrated T cells. Supplementary Information The online version contains supplementary material available at 10.1186/s13045-021-01120-3.

Chimeric antigen receptor T-cell (CAR T) therapy is a kind of effective cancer immunotherapy. However, designing CARs remains a challenge because many targetable antigens are shared by T cells and tumor cells. This shared expression of antigens can cause CAR T cell fratricide. CD38-targeting approaches (e.g., daratumumab) have been used in clinical therapy and have shown promising results. CD38 is a kind of surface glycoprotein present in a variety of cells, such as T lymphocytes and tumor cells. It was previously reported that CD38-based CAR T cells may undergo apoptosis or T cell-mediated killing (fratricide) during cell manufacturing. In this study, a CAR containing a sequence targeting human CD38 was designed to be functional. To avoid fratricide driven by CD38 and ensure the production of CAR T cells, two distinct strategies based on antibodies (clone MM12T or clone MM27) or proteins (H02H or H08H) were used to block CD38 or the CAR single-chain variable fragment (scFv) domain, respectively, on the T cell surface. The results indicated that the antibodies or proteins, especially the antibody MM27, could affect CAR T cells by inhibiting fratricide while promoting expansion and enrichment. Anti-CD38 CAR T cells exhibited robust and specific cytotoxicity to CD38+ cell lines and tumor cells. Furthermore, the levels of the proinflammatory factors TNF-α, IFN-γ and IL-2 were significantly upregulated in the supernatants of A549CD38+ cells. Finally, significant control of disease progression was demonstrated in xenograft mouse models. In conclusion, these findings will help to further enhance the expansion, persistence and function of anti-CD38 CAR T cells in subsequent clinical trials.

Coastal wetlands in East China are essential stopover places for birds along the East Asian-Australian Flyway. However, numerous wind turbines have been built in or near these wetlands in recent years, which might disturb the bird community in the area. Therefore, investigating the bird community and its responses to wind farms in coastal wetlands of East China is of great significance for bird conservation. In the spring and autumn of 2019 and 2020, we investigated the bird community in the Rudong coastal wetland in East China using point counts. We determined 4 geographical factors at each census point, i.e., distance to the wind farm boundary (DW), distance to the suburbs, distance to the sea, and vegetation area, and analysed the relationship between bird number and DW through partial correlation analysis. A total of 11 orders and 103 species of birds, including 4 endangered species, were observed during our survey. Charadriiformes was the dominant taxon in the wetland, and Calidris alpina was the most common species in both spring and autumn. Passeriformes exhibited high species richness but low numbers. The results of partial correlation analysis indicated that birds’ responses to the wind farm varied depending on their dominance and category: dominant and subdominant birds tended to avoid the wind farm, whereas rare birds tended to approach them; aquatic birds were alert to the wind farm, whereas terrestrial birds better adapted to them. We concluded that the dominant aquatic birds, including the endangered species Calidris tenuirostris, were most negatively impacted by the wind farm; the occasional birds and rare aquatic birds might be disturbed by wind farm but not significantly so; and the rare terrestrial birds were least disturbed by or even benefited from the wind farm.

In order to develop biodegradable phase change materials (PCMs) with high latent heat for cold chain logistics, superabsorbent resin (SAR) was prepared based on starch graft copolymerization. FTIR and DSC demonstrated that acrylic acid was successfully grafted onto starches and optimum latent heat of PCM was 330.4 J/g with 10% (w/w) starch. The water retention of PCM with 10% (w/w) starch was 0.49 after heating at 50 °C for 200 h, which was 4.9 folds higher than that of non-starch PCM. Biodegradation rate of PCM was 60.12% within 75-day burial, which was 6 folds higher than that of non-starch PCM. Moreover, significant reduction in browning index, odor, decay, relative conductivity and malondialdehyde (MDA) content was observed in Lentinus edodes treated by biodegradable PCM. These results indicated that the application of biodegradable PCM could extend the shelf life of fresh L. edodes stored at 25 °C.

Mechanical damage and microbial infestation were critical issues during strawberry transportation. This study prepared a modified polyvinyl formal (PVFM) vibration-damping material and investigated its application in strawberry transportation. This modified PVFM was primarily composed of polyvinyl alcohol, starch, formaldehyde, and allyl isothiocyanate. Our results showed that the modified PVFM had good vibration-damping and antifungal properties with 0.137 g/m3 of density, a water absorption ratio value of 6.24, 278 Pa of tensile strength, 3.86 Pa of elastic modulus, a buffer coefficient of 0.318, and antifungal rates of 68.8% and 80.6% against Bacillus anthracis and Botrytis cinerea, respectively. Moreover, this modified PVFM could enhance the storage life of strawberry by improving PAL activity from 376 to 3,125 U/g, inhibiting POD activity with a maximum value of 3.1 U/g, and decreasing CAT activity from 324 to 35 U/g. The modified PVFM presented a good vibration-damping capacity and the potential to extend strawberry shelf life. PRACTICAL APPLICATIONS: Berry fruits were vulnerable to mechanical damage caused by vibration during transportation, resulting in quality deterioration, rot, and aging. In this study, modified polyvinyl formal (PVFM) was prepared and was of low density and displayed good mechanical, vibration-damping, and antifungal properties. The modified PVFM could improve the storage life of strawberry. Thus, the modified PVFM could have potential application in berry fruits transportation.

Small-scale aquaculture operation is being increasing practised around the world, particularly in the developing countries, but the greenhouse gas (GHG) dynamics and fluxes from small aquaculture ponds are still poorly assessed. In this study, the dissolved concentrations and fluxes of CO2 and CH4 were determined in three coastal earthen shrimp ponds over one whole year, covering both the farming and non-farming periods, in the Min River Estuary, southeastern China. Both ebullitive and diffusive CH4 fluxes were measured, which were rarely done in previous studies. The average concentrations of dissolved CO2 and CH4 in water column in the farming period varied between 18.1 ± 0.1 and 79.6 ± 1.1 μmol L−1, and 1.3 ± 0.1 and 55.9 ± 3.2 μmol L−1, respectively. When averaged across the whole year, the mean CO2 and CH4 fluxes from the ponds were − 18.4 ± 7.4 and 22.6 ± 6.9 mg m−2 h−1, respectively, suggesting that the shrimp ponds acted as a CO2 sink and a CH4 source. Based on the sustained-flux global warming potential (SGWP) and sustained-flux global cooling potential (SGCP) models, the annual warming potential was estimated to be 7.1 × 103 g CO2-eq m−2 yr−1, with approximately 90% of this arising from the farming period. Ebullition was the dominant emission pathway for CH4, accounting for over 90% of the total CH4 emission during the farming period. This full-year study improves our understanding of carbon cycling in coastal aquaculture ponds and provides a scientific basis for updating the GHG inventories.

The tidal estuarine wetlands of China are rich in plant diversity, but several human-driven processes, such as species invasion, can affect the biogeochemical cycles of these ecosystems, and by changing soil conditions can inhibit the regeneration of native vegetation. We seasonally analyzed the carbon (C), nitrogen (N) and phosphorus (P) concentrations in soils and in leaves, stems and roots of the invasive species Spartina alterniflora and of the native species Cyperus malaccensis var. brevifolius Boeckeler. This latter species was analyzed both in natural non-invaded stands and in stands that had been invaded by Spartina but from which it had been removed and replaced by Cyperus. The aim was to investigate the effect of plant invasion, subsequent removal and replanting with a native species on C, N and P stoichiometry of the plant–soil system in the tidal wetlands of the Minjiang River. C and N concentrations averaged across seasons did not differ significantly among the plant species. P concentration was lower in the stems of Spartina than in the stems of the native species Cyperus but was not significantly different in the roots of the two species. The soil C and N concentrations were higher in the Spartina stand than in the Cyperus stand, whereas the soil P concentrations were not significantly different. The invasive species had a higher N-resorption capacity, N:P ratios in stem and roots, biomass, absolute growth and biomass N and had a lower relative growth rate and litter production than the native species. After the removal of the invasive plants, the regenerating native plants have a higher capacity to resorb N and lower relative growth rates. All these traits show that a conservative strategy and a high N-use efficiency and internal plant control of the N in the ecosystem underlie the invasive success of Spartina in this N-limited wetland. Relative growth rate was associated with lower plant N:P ratios, whereas absolute growth rate was associated with higher nutrient-use efficiency and lower C and N turnover and storage capacities in the biomass. Changes in soil properties produced by the establishment of an invasive plant can condition the later regeneration of native plants.

Background and aims: Nitrogen (N) and phosphorus (P) are essential nutrients for plant growth, and their availability and stoichiometry play pivotal roles in trophic dynamics and community composition. The biogeochemical niche (BN) hypothesis claims that each species should have an optimal elemental composition and stoichiometry as a consequence of its optimal function in its specific ecological niche. Little attention, however, has been given to N and P stoichiometric patterns and test the BN hypothesis in coastal wetland communities from the perspective of organ and species-specific comparisons.Methods: We investigated factors responsible for changes in N and P stoichiometry patterns in different functional groups in coastal wetlands and tested the BN hypothesis by evaluating N and P composition in whole aboveground plants and organs.Results: Both plant N and P concentrations were high in coastal wetlands, indicating that N and P were not likely limiting, although the N:P ratio was slightly lower than the ratio reported in global and Chinese terrestrial flora. N and P concentrations and N:P ratios varied strongly between C₃ and C₄ species, among species, and among organs within species. N and P concentrations were not correlated with latitude, mean annual temperature and precipitation, although N:P ratio was weakly correlated with these factors. The differences in N and P concentrations and N:P ratios along the wetland gradients were mainly because of the species-specific community composition of each site.Conclusions: The results are consistent with the BN hypothesis. First, N and P composition is species-specific (homeostatic component of BN), each species tends to maintain its own composition even growing in different sites with different species composition. Second, different species, despite maintaining their own composition, have distinct degree of composition phenotypic flexibility (flexibility component of BN); this different size of "biogeochemical space" was observed when comparing different species living in the same community and the shifts in species BN space and size was observed when comparing populations of the same species living in different sites.

Background Anti-CD19-directed chimeric antigen receptor (CAR) T-cell therapy has had a resounding effect on the treatment of B-ALL. However, CAR T cells have been less effective against B-cell non-Hodgkin lymphoma (B-NHL), in part because they become a exhausted state triggered by chronic antigen stimulation and characterized by upregulation of inhibitory receptors and loss of effector function.1-4 It has recently been demonstrated that de novo DNA methylation promoted T-cell exhaustion, whereas methylation inhibition enhanced ICB-mediated T-cell rejuvenation in vivo.5 6 FDA-approved DNA demethylating agents, such as decitabine (DAC), may provide a means to modify exhaustion-associated DNA methylation programs that restrict ICB-responsiveness. Methods We treated CAR (CAR-CD19-expressing) T cells with low-dose DAC (dCAR T cells), to determine its effects on antitumor activities, exhaustion- and memory-associate cell phenotype change, cell cytokine production, and cell proliferation. Its impact on antitumor activities was evaluated in vitro functional assays and mouse in vivo studies. We also conducted western blot, flow cytometry, methylation analysis, RNA in situ hybridization and high throughput RNA sequencing to determine the underlying mechanisms of dCAR T cell function. Results The low-dose, short-term DAC treatment in vitro enhanced the central memory (Tcm) population and the ration of CD4/CD8, and induced degradation of DNMT3a.CAR T cell treated by DAC developing into less-differention status by enhancing memory. dCAR T cells exhibit enhanced antitumour reactivity and the maintenance of a memory-like phenotype at low effector:target ratios. Especially shown by the ‘stress test’, the dCAR T cells at very low doses could efficiently control tumours with a very large burden, and have effective recall responses upon tumour re-challenge in vivo. Importantly, the dCAR T cells maintained a higher proportion of cells with a memory phenotype than did the CAR T cells under long-term tumour stimulation. Transcription of gene sets involved in memory maintenance, proliferation, cytokine production and anti-inhibitor processes was triggered by antigen-expressing target cells upon DAC exposure before antigen stimulation. dCAR T cells avoided the exhaustion programme induced during tumour cell stimulation; they did not upregulate the expression of genes encoding inhibitory receptors and retained relatively high expression of memory related transcription factors and genes. Conclusions CAR T cells underwent DNA reprogramming after DAC treatment, which induced significant sustained cell expansion, cytotoxicity, and cytokine production and reduced exhaustion after antigen exposure. Acknowledgements We thank Professor Lin Xin of Tsinghua University and Professor Mingzhou Guo of Chinese PLA General Hospital for support of data analysis. References Wherry EJ, Kurachi M. Molecular and cellular insights into T cell exhaustion. Nat Rev Immunol 2015;15:486–499. doi:10.1038/nri3862 Wherry EJ, et al. Molecular signature of CD8+ T cell exhaustion during chronic viral infection. Immunity 2007;27:670–684. doi:10.1016/j.immuni.2007.09.006 Schietinger A, et al. Tumor-specific T cell dysfunction is a dynamic antigen-driven differentiation program initiated early during tumorigenesis. Immunity 2016;45:389–401. doi:10.1016/j.immuni.2016.07.011 Schietinger A, Greenberg PD. Tolerance and exhaustion: defining mechanisms of T cell dysfunction. Trends Immunol 2014;35:51–60. doi:10.1016/j.it.2013.10.001 Ghoneim HE, et al. De novo epigenetic programs inhibit PD-1 blockade-mediated T cell rejuvenation. Cell 2017;170:142–157.e119. doi:10.1016/j.cell.2017.06.007 Pauken KE, et al. Epigenetic stability of exhausted T cells limits durability of reinvigoration by PD-1 blockade. Science 2016;354:1160–1165. doi:10.1126/science.aaf2807

Chimeric antigen receptor T (CAR T) cells targeting CD19 have achieved breakthroughs in the treatment of hematological malignancies, such as relapsed/refractory non-Hodgkin lymphoma (r/rNHL); however, high rates of treatment failure and recurrence after CAR T cell therapy are considerable obstacles to overcome. In this study, we designed a series of tandem CARs (TanCARs) and found that TanCAR7 T cells not only showed dual antigen targeting of both CD19 and CD20 but also formed superior and stable immunological synapse (IS) structures, which may be related to their robust antitumor activity. In an open-label, single-arm phase I/IIa trial (ClinicalTrials.gov number NCT03097770), we enrolled 33 patients with r/rNHL, and a total of 28 patients received an infusion after conditioning chemotherapy. The primary objective was to evaluate the safety and tolerability of TanCAR7 T cells. Efficacy, progression-free survival and overall survival were evaluated as secondary objectives. Cytokine release syndrome (CRS) occurred in 14 patients (50%), with 36% grade 1 or 2 and 14% grade 3. No cases of CAR T cell-related encephalopathy syndrome (CRES) of grade 3 or higher were confirmed in any patient. One patient died from a treatment-associated severe pulmonary infection. The overall response rate was 79% (95% confidence interval [CI], 60 to 92), and the complete response rate was 71%. The progression-free survival rate at 12 months was 64% (95% CI, 43 to 79). In this study, TanCAR7 T cells elicited a potent and durable antitumor response but not grade 3 or higher CRES in patients with r/rNHL.

Background Despite the impressive complete remission (CR) induced by CD19 CAR-T cell therapy in B-ALL, the high rate of complete responses is sometimes limited by the emergence of CD19-negative leukemia. Bispecific CAR-modified T cells targeting both CD19 and CD22 may overcome the limitation of CD19-negative relapse. Methods We here report the design of a bispecific CAR simultaneous targeting of CD19 and CD22. We performed a phase 1 trial of bispecific CAR T cell therapy in patients with relapsed/refractory precursor B-ALL at a dose that ranged from 1.7 × 106 to 3 × 106 CAR T cells per kilogram of body weight. Results We demonstrate bispecific CD19/CD22 CAR T cells could trigger robust cytolytic activity against target cells. MRD-negative CR was achieved in 6 out of 6 enrolled patients. Autologous CD19/CD22 CAR T cells proliferated in vivo and were detected in the blood, bone marrow, and cerebrospinal fluid. No neurotoxicity occurred in any of the 6 patients treated. Of note, one patient had a relapse with blast cells that no longer expressed CD19 and exhibited diminished CD22 site density approximately 5 months after treatment. Conclusion In brief, autologous CD19/CD22 CAR T cell therapy is feasible and safe and mediates potent anti-leukemic activity in patients with relapsed/refractory B-ALL. Furthermore, the emergence of target antigen loss and expression downregulation highlights the critical need to anticipate antigen escape. Our study demonstrates the reliability of bispecific CD19/CD22 CAR T cell therapy in inducing remission in adult patients with relapsed/refractory B-ALL. Trial registration ClinicalTrials.gov identifier: NCT03185494.

The cuticular wax layer in fruit is a hydrophobic barrier which protects fruit from biotic and abiotic stress. The cuticular wax also affect fruit quality. This paper investigated the effects of salicylic acid on fruit quality and the cuticular wax in blueberry fruit during storage at room temperature (25 °C). ‘Powderblue’ blueberries (Vaccinium virgatum Ait) were treated with 1.0 mmol L-1 salicylic acid. The composition of cuticular wax layer and structure of epicuticular wax layer were analyzed at 4 d intervals during storage. Salicylic acid could efficiently delay the reduction of total wax content and affected the proportions of its constituents, including triterpenoids, esters, sterols, fatty acids, alcohols and alkanes. Our results also showed that there was no significant difference on the structure of epicuticular wax in salicylic acid treated fruits compared with that of the control. Salicylic acid delayed deterioration of blueberry fruit and enhanced its resistance to disease during storage.

Conductive materials/minerals can promote direct interspecies electron transfer (DIET) between syntrophic bacteria and methanogens in defined co-culture systems and artificial anaerobic digesters; however, little is known about the stimulation strategy of carbon material on methane production in natural environments. Herein, the effect of carbon cloth, as a representative of conductive carbon materials, on methane production with incubated wetland soil was investigated. Carbon cloth significantly promoted methanogenesis. With the application of electrochemical technology, calculation of the apparent electron transfer rate constant showed that carbon cloth significantly increased electron transfer rate (ETR) compared with the control experiment in presence of cotton cloth, from 0.0017 ± 0.0003 to 0.0056 ± 0.0015 s−1. Results obtained from both stable carbon isotope measurements and application of specific inhibitor (CH3F) for acetoclastic methanogenesis indicated that carbon cloth obviously promoted acetoclastic methanogenesis instead of CO2 reduction. High-throughput sequencing showed that methane production may stem from the involvement of Methanosarcina for both treatments. Our findings suggested that conductive carbon material can promote acetoclastic methanogenesis instead of CO2 reduction in a natural environment.

The increasing success of invasive plant species in wetland areas can threaten their capacity to store carbon, nitrogen, and phosphorus (C, N, and P). Here, we have investigated the relationships between the different stocks of soil organic carbon (SOC), and total C, N, and P pools in the plant-soil system from eight different wetland areas across the South-East coast of China, where the invasive tallgrass Spartina alterniflora has replaced the native tall grasses Phragmites australis and the mangrove communities, originally dominated by the native species Kandelia obovata and Avicennia marina. The invasive success of Spartina alterniflora replacing Phragmites australis did not greatly influence soil traits, biomass accumulation or plant-soil C and N storing capacity. However, the resulting higher ability to store P in both soil and standing plant biomass (approximately more than 70 and 15 kg P by ha, respectively) in the invasive than in the native tall grass communities suggesting the possibility of a decrease in the ecosystem N:P ratio with future consequences to below- and aboveground trophic chains. The results also showed that a future advance in the native mangrove replacement by Spartina alterniflora could constitute a serious environmental problem. This includes enrichment of sand in the soil, with the consequent loss of nutrient retention capacity, as well as a sharp decrease in the stocks of C (2.6 and 2.2 t C ha-1 in soil and stand biomass, respectively), N, and P in the plant-soil system. This should be associated with a worsening of the water quality by aggravating potential eutrophication processes. Moreover, the loss of carbon and nutrient decreases the potential overall fertility of the system, strongly hampering the reestablishment of woody mangrove communities in the future.

Ecosystem respiration (Reco) is an important pathway for gaseous carbon loss in coastal wetland environments. However, little is known regarding the dynamics of Reco and its temperature sensitivity (Q10) under different tidal stages, and vegetation types. In this study, Reco was measured with dark chambers for nearly 2 years in three different brackish marsh stands dominated by Cyperus malaccensis, Phragmites australis, and Spartina alterniflora, respectively, in the Min River estuary of southeast China. The mean Reco across tidal stages and vegetation types exhibited strong temporal variations, and the Q10 value for Reco was higher in the warm months than in the cold months. The mean annual Reco was negatively correlated with electrical conductivity and soil water content, but positively correlated with both tidal water level and soil pH (P 0.05). Overall, the C. malacensis and S. alterniflora stands had the highest and lowest average rates of Reco, respectively, while the mean Qs10 values were lower in the Phragmites australis stand than the other two vegetation types. Our findings point to the need of strengthening the in situ measurements of gas fluxes at multiple time scales in order to develop a better understanding of the response of Reco to changing environmental conditions in the subtropical coastal wetlands. Our results also reiterate the importance of considering the influence of vegetation types in characterizing the magnitude and temperature sensitivity of Reco in estuarine tidal marshes for predicting ecosystem responses to future global change.

Although the role of microbial iron respiration in tidal marshes has been recognized for decades, the effect of rhizosphere processes on dissimilatory ferric iron reduction (FeR) is poorly known. Herein, we examined the FeR surrounding the root zone of three tidal marsh plants. Using in situ rhizoboxes, we accurately separated rhizobox soil as one rhizosphere zone, and three bulk soil zones. Dissimilatory and sulfidic-mediated FeR were quantified by accumulation of non-sulfidic Fe(II) and Fe sulfides over time, respectively. The rates of dissimilatory FeR attained 42.5 μmol Fe g−1 d−1 in the rhizosphere, and logarithmically declined by up to 19.1 μmol Fe g−1 d−1 in the outer bulk soil. The rates of sulfidic-mediated FeR were less than 2 μmol Fe g−1 d−1 among all zones. Poorly crystalline Fe(III), DOC and DON, porewater Fe2+, and SO42− were all enriched in the rhizosphere, whereas non-sulfidic Fe(II) and Fe sulfides gradually accumulated away from the roots. Iron reducers (Geobacter, Bacillus, Shewanella, and Clostridium) had higher populations in the rhizosphere than in the bulk soil. Higher rates of dissimilatory FeR were observed in the Phragmites australis and Spartina alterniflora rhizoboxes than in the Cyperus malaccensis rhizoboxes. The radial change pattern of dissimilatory FeR rates were determined by allocation of poorly crystalline Fe(III) and dissolved organic carbon. The interspecies difference of rhizosphere dissimilatory FeR was associated with the root porosity and aerenchyma of the tidal marsh plants.

Psoriasis is a chronic inflammatory skin disorder mediated by the cells and molecules of both the innate and adaptive immune systems. Autologous cytokine-induced killer (CIK) cell infusion is considered an effective and safe cancer treatment and is licensed for this use in China. Accumulated evidence indicating that CD3+CD56+ cells are significantly decreased in psoriatic patients prompted us to investigate if the restoration of CD3+CD56+ cells may be beneficial for psoriatic patients. We designed a clinical trial for psoriasis treatment that involved CIK cell infusion because CIK cells include a large amount of CD3+CD56+ T cells (NCT01894373 at www.clinicaltrials.gov). Six patients with severe psoriasis were initially enrolled, and four of them exhibited markedly lower levels of CD3+CD56+ cells in their peripheral blood (PB) relative to healthy donors. CIK cell infusion-associated toxicity was not observed in any infusion. The percentage of CD3+CD56+ cells in the PB markedly increased and the psoriasis area and severity index (PASI) synchronously decreased in four patients with lower CD3+CD56+ cell contents, and two of them obtained a more than 4-month PASI75 after completing a four-cycle treatment. However, a decrease in the CD3+CD56+ cells was observed concomitantly with disease recurrence after short-term amelioration. In contrast, no obvious improvement was observed in the two patients with nearly normal CD3+CD56+ cells in the PB before treatment. These observations suggest that the normalization of the CD3+CD56+ cell level may improve the skin lesions of severe psoriasis and warrant further clinical trials for severe psoriasis using repeated CIK adoptive immunotherapy.

Tidal marshes play an important functional role in removing nitrogen (N) pollution before delivery to coastal and ocean systems; however, little is known about their removal capacity as N2O gas emissions from different plant species. To evaluate the effects of N inputs on N2O emissions from tidal marshes, we measured N2O fluxes from native (Cyperus malaccensis) and invaded (Spartina alterniflora) tidal marshes in the Min River estuary, and fertilized with exogenous N at the rates of 0, 21 and 42 g N m−2 yr.−1, respectively. S. alterniflora invasion did not significantly influence N2O emissions from the C. malaccensis marsh under natural conditions, but under N addition conditions, the invasion of S. alterniflora decreased N2O emissions, primarily owing to its stronger N uptake capacity. Exogenous N had significant positive effects on N2O fluxes in both native and invaded tidal marshes. Moreover, significant temporal variability of N2O fluxes was observed after N was gradually added to the native and invaded marshes. Within 3 h of N addition, N2O fluxes were significantly higher in plots receiving N additions relative to controls. After 8 days, few significant differences were found between treatments. Moreover, electrical conductivity, pH and oxidation-reduction potential at different soil depths were not significantly affected by N addition. Considering N addition showed extremely high positive effects on N2O fluxes at the hours scale, the overall increase of N2O emissions from wetlands in response to N addition may be significantly underestimated. To better assess the global climatic role of salt marshes that have been affected by N addition, the short-term temporal variability of N2O emissions should receive greater attention.

Bacterial community composition influences the formation of flavourful substances in fermented foods. However, the bacterial structure of fermented vegetables from different origins has scarcely been reported. Here, 10 different fermented vegetables of Chinese origin were collected to explore the bacterial community diversity using 16S rDNA sequencing. Of these samples, the number of identified operational taxonomic units (OTUs) ranged from 107 to 290, which could be categorized into 12 phyla, 21 classes, 52 orders, 97 families, 223 genera and 348 species. Firmicutes and Proteobacteria were the top two dominant phyla with a relative abundance of 88.2–99.3% of the total OTUs. Meanwhile, Lactobacillus alimentarius , L. versmoldensis , Bacillus and unclassified Weissella spp. were detected as the predominant Firmicutes bacteria, while Chromohalobacter japonicus , Halomonas jeotgali , Pediococcus and unclassified Vibrio spp. were the most abundant proteus bacteria. Hierarchical cluster and principal component analysis indicated that the 10 samples could be grouped into three groups, with the main two components explaining 49.1 and 28.6% of the total variance. These results indicated a high level of diversity in the bacterial community structure in traditional fermented vegetables and demonstrated the possible influences of manufacturing location and conditions, and/or the type of raw material, on bacterial diversity.

Invasion of an exotic C 4 plant Spartina alterniflora has been shown to increase soil organic carbon (SOC) concentrations in native C 3 plant-dominated coastal wetlands of China. However, little is known about the effects of S. alterniflora invasion on SOC concentrations and fractions in tidal marshes dominated by native C 4 plants. In this study, a field experiment was conducted in a tidal marsh dominated by the native C 4 plant Cyperus malaccensis in the Minjiang River estuary, China. Concentrations of SOC and liable SOC fractions, dissolved organic carbon (DOC), microbial biomass carbon (MBC), and easily oxidizable carbon (EOC), were measured in the top 50-cm soils of the C. malaccensis community, as well as those of three S. alterniflora communities with an invasion duration of 0–4 years (SA-4), 4–8 years (SA-8), and 8–12 years (SA-12), respectively. Results showed that both SOC stocks in the 50-cm soils and mean SOC concentrations in the surface soils (0–10 cm) of the C. malaccensis community increased with the duration of S. alterniflora invasion, whereas SOC concentrations in the 10–50-cm soils decreased slightly during the initial period of S. alterniflora invasion, before increasing again. The pattern of changes in labile SOC fractions (DOC, MBC, and EOC) with invasion duration was generally similar to that of SOC, while the ratios of labile SOC fractions to total SOC (DOC:SOC, MBC:SOC, and EOC:SOC) decreased significantly with the duration of S. alterniflora invasion. The findings of this study suggest that invasion of the exotic C 4 plant S. alterniflora into a marsh dominated by the native C 4 plant C. malaccensis would enhance SOC sequestration owing to the greater amount of biomass and lower proportion of labile SOC fractions present in the S. alterniflora communities.

Methane emissions from wetlands contribute to the currently rising levels of methane in the atmosphere. Nonetheless, the assessments of methane-producing Archaea (methanogens) and their environmental controls are still rare particularly in soils from estuarine tidal marshes. In estuarine tidal marshes, the salinity gradient is commonly observed, and its effects on the methanogenic community are hardly known. In this study, in the Min River estuary, southeast China, we analyzed two freshwater and two brackish (oligohaline) marshes in a subtropical estuary, where we assessed methanogenic community structure and abundance by molecular cloning, sequencing, and quantitative PCR methods, as well as by means of soil physicochemical characteristics and concentration of methane dissolved in pore water. The common and dominant genera of these methanogens include Methanoregula, Methanobacterium, Methanosarcina, and Methanolinea in the four marshes. Phylotypes of the methanogens using H2/CO2 (Methanomicrobiales) or acetate (Methanosarcinales) were identified in the mcrA gene clone libraries and by restriction fragment length polymorphism (RFLP) analyses. The abundance levels of methanogens in the two freshwater marshes were significantly higher than those in the two brackish marshes. The value of the Shannon Index (H′) of diversity and Simpson index (D) of evenness of methanogen mcrA clone libraries decreased along the salinity gradient from freshwater to brackish water. Methanogen community structure varied with soil depth in the four wetlands. The concentrations of methane in pore water also decreased from the freshwater sites to the brackish sites; this pattern is consistent with a change in the abundance of methanogens. Our results showed that in a low-level salinity gradient of a subtropical estuary, salinity is still a major factor controlling the overall community structure and abundance of methanogens.

We measured methane emissions together with acetate, DMS, sulfate, and dissolved methane concentrations in pore waters from adjacent marsh plant communities in an oligohaline marsh of the Min River estuary, southeastern China—one community dominated by a native species, Cyperus malaccensis, the other by the exotic Spartina alterniflora. The objective was to determine if the exotic species altered soil pore-water chemistry, metabolite pools, and methane emissions. We found that the invasive S. alterniflora, significantly increased the concentration of pore-water acetate (6.1 vs. 1.2 μM), but did not increase the pools of pore-water DMS (54.6 vs. 45.9 nM). We observed significantly greater annual emissions of methane from the marsh areas dominated by S. alterniflora (15.1 vs. 5.1 mg m−2 h−1 annual average) and a significant correlation between annual methane emission rate and pore-water acetate concentration at soil depths between 1 cm and 20 cm. There was no significant correlation between methane emission and the concentration of pore-water DMS in either marsh community. The results indicate that a change in the dominant plant community can alter soil biogeochemistry. In particular, in an oligohaline environment, S. alterniflora supported higher rates of methane production than the native plant community. It probably does so through a variety of metabolic pathways, probably including acetoclastic methanongenesis.

Human epidermal growth factor receptor 2 (HER2) proteins are overexpressed in a high proportion of gastric cancer (GC) cases and affect the maintenance of cancer stem cell (CSC) subpopulations, which are used as targets for the clinical treatment of patients with HER2-positive GC. Despite improvements in survival, numerous HER2-positive patients fail treatment with trastuzumab, highlighting the need for more effective therapies. In this study, we generated a novel type of genetically modified human T cells, expressing a chimeric antigen receptor (CAR), and targeting the GC cell antigen HER2, which harbors the CD137 and CD3ζ moieties. Our findings show that the expanded CAR-T cells, expressing an increased central memory phenotype, were activated by the specific recognition of HER2 antigens in an MHC-independent manner, and effectively killed patient-derived HER2-positive GC cells. In HER2-positive xenograft tumors, CAR-T cells exhibited considerably enhanced tumor inhibition ability, long-term survival, and homing to targets, compared with those of non-transduced T cells. The sphere-forming ability and in vivo tumorigenicity of patient-derived gastric cancer stem-like cells, expressing HER2 and the CD44 protein, were also inhibited. Our results support the future development and clinical application of this adoptive immunotherapy in patients with HER2-positive advanced GC. Electronic supplementary material The online version of this article (doi:10.1007/s13238-017-0384-8) contains supplementary material, which is available to authorized users.

Despite the growing recognition regarding the carbon cycle in the rhizosphere of upland ecosystems, little is known regarding the rhizosphere effect on soil organic carbon (SOC) mineralization in tidal marsh soils. In the current study, in situ rhizobox experiments (including rhizosphere and inner and outer bulk soil) were conducted in an estuarine tidal marsh. Our results showed that a higher abundance of total bacteria, Geobacter, dsrA and mcrA and lower α-diversity were observed in the rhizosphere relative to the bulk soil. Rhizosphere effects shifted the partition of terminal metabolic pathways from sulfate reduction in the bulk soil to the co-dominance of microbial Fe(III) and sulfate reduction in the rhizosphere. Although the rhizosphere effect promoted the rates of three terminal metabolic pathways, it showed greater preference towards microbial Fe(III) reduction in the tidal marsh soils. Plant species had little impact on the partitioning of terminal metabolic pathways, but did affect the potential of total SOC mineralization together with the abundance and diversity of total bacteria. Both the rhizosphere effect and plant species influenced the bacterial community composition in the tidal marsh soils; however, plant species had a less pronounced impact on the bacterial community compared with that of the rhizosphere effect.

Background Chimeric antigen receptor T (CAR-T) cell therapy simultaneously against CD19 and CD22 is an attractive strategy to address the antigen escape relapse after CD19-directed CAR-T cell therapies. However, the potential of optimizing the durability of remission by this approach in patients with B cell acute lymphoblastic leukemia (B-ALL) remains a critical unanswered question so far. Case presentation We treated an adult patient with relapsed and refractory B-ALL after haploidentical hematopoietic stem cell transplantation (HSCT) by administering haploidentical CAR-T cells targeting both CD19 and CD22 following preparative lymphodepleting chemotherapy. This patient has remained in minimal residual disease-negative remission for more than 14 months and has been tapered off graft versus host disease prophylaxis. Conclusions CAR simultaneously targeting CD19 and CD22 has the potential of inducing long-term remission in patients with B-ALL. Electronic supplementary material The online version of this article (10.1186/s13045-019-0741-6) contains supplementary material, which is available to authorized users.

Rice lipids are a major class of nutritional components in rice grain, having relative significance on rice grain quality. In this chapter, the main class and concentration of natural lipids in rice grain, as well as their relations with aroma, flavor, hardness, storage, milling, eating, and cooking qualities are systemically reviewed. The genetic and environmental factors have both positive and negative influences on native lipid distribution and function in rice. Numerous rice lipids synthesis related genes and quantitative trait loci (QTLs) are summarized for understanding of the genetic basis and metabolism of lipid biosynthesis in rice grain. Rice bran oil (RBO) as a valuable by-product in rice industry is one of the healthiest edible oils. The primary bioactive ingredients such as γ-oryzanol in RBO and their health-promoting effects are also discussed. The available methods for RBO extraction, analysis and processing, and variations in bioactive phytochemicals in RBO due to these different methods are compared. This chapter may enhance our understanding of the importance of lipids in rice grain and provide insights into the potential applications in the rice industry.

Background: Despite the impressive CR induced by CD19 CAR-T cell therapy in B-ALL, the high rate of complete responses is sometimes limited by the emergence of CD19-negative leukemia. Bispecific CAR-modified T cells targeting both CD19 and CD22 may overcome the limitation of CD19-negative relapse. Methods: Here, we report the design of a bispecific CAR that triggers robust cytolytic activity against target cells by targeting either CD19 or CD22. We performed a phase 1 trial of bispecific CAR T cell therapy in patients with relapsed/refractory precursor B-ALL at a dose that ranged from 1.7×106 to 3×106 CAR T cells per kilogram of body weight. Finding: MRD-negative CR was achieved in 6 out of 6 enrolled patients. Autologous CD19/CD22 CAR T cells proliferated in vivo and were detected in the blood, bone marrow, and cerebrospinal fluid. No neurotoxicity occurred in any of the 6 patients treated. Of note, one patient had a relapse with blast cells that no longer expressed CD19 and exhibited diminished CD22 site density approximately 5 months after treatment. Interpretation: In brief, autologous CD19/CD22 CAR T cell therapy is feasible and safe and mediates potent antileukemic activity in patients with relapsed/refractory B-ALL. Furthermore, the emergence of target antigen loss and expression downregulation highlights the critical need to anticipate antigen escape. Our study demonstrates the reliability of bispecific CD19/CD22 CAR T cell therapy in inducing remission in adult patients with relapsed/refractory B-ALL. Trial Registration: NCT03185494. Funding Statement: National Key Research and Development Program of China and National Natural Science Foundation of China. Declaration of Interests: The authors declare that they have no competing interests. Ethics Approval Statement: The study protocols were approved by the institutional review board at the Chinese PLA general hospital, and the patients provided written informed consent. This clinical investigation was conducted according to the principles of the Declaration of Helsinki.

Wetlands play an important role in the global carbon cycle but are intensively disturbed by humans. Estimating the gaseous carbon budget of degraded wetlands has great significance for wetland conservation. The Shanyutan wetland is a degraded tidal wetland in Southeast China. We compared the landscape patterns of Shanyutan in 2009 and 2019 based on remote sensing images and determined the sediment CO2 flux (excluding root respiration) in various land types. We combined the sediment CO2 flux with the wetland CH4 flux, and the CH4 and CO2 fluxes of aquaculture ponds, as well as net plant carbon sequestration (NPCS) from previous studies, and calculated the gaseous carbon budget of Shanyutan for the 10-year period. Our results show that the areas of aquaculture ponds and mudflats increased over the 10-year period, whereas those of vegetated marsh and sand decreased. Determination of the sediment CO2 flux indicates that sediment CO2 flux is higher in vegetated marsh and lower in bare land, with the highest annual flux in Spartina alterniflora marsh and the lowest in sand. The gaseous carbon budget shows that aquaculture ponds and bare land are sources of atmospheric carbon, whereas vegetated marsh serves as a sink. The entire wetland was an atmospheric carbon sink in 2009, with a carbon sequestration rate of 1364.4 tC·a−1; however, it became a source in 2019 due to destruction, with a carbon efflux of 1072.6 tC·a−1. Moreover, the gaseous carbon balance in Shanyutan had a climate-warming potential, with an equivalent CO2 production of 5971.8 t·a−1 in 2009, and this potential increased ~9 times over the 10-year period. Finally, we conclude that restoring the carbon cycle of Shanyutan has considerable future economic value.

Assessing the diverse communities of methanogenic Archaea and sulfate-reducing bacteria (SRB) is important to understand methane (CH4) production in wetland ecosystems. However, the vertical distribution of composition and diversity, and the effects of environmental factors on the methanogen and SRB communities in the sediments of subtropical estuarine brackish marshes have been poorly characterized. To assess the effects of variable environmental conditions on methanogenic and SRB communities in marshes, we studied three brackish marsh zones dominated by Phragmites australis, Cyperus malaccensis and Spartina alterniflora, respectively, in the Min River estuary, southeastern China. Methanogens of the Methanomicrobiales order was the dominant group at sediment depths of 0–30 cm, which indicated that the main pathway of methane production was H2/CO2 in this zone. In general, methanogens of the genus Methanoregula were dominant in the three marsh zones. For SRB, Desulfobacterales was the dominant group, and Desulfobacterium and Desulfosarcina were the predominant genera at the depth of 0–30 cm. The community composition of methanogens and SRB changed with vegetation type and soil depth. Compared with SRB, vegetation type demonstrated a stronger influence on the community composition of methanogens. Canonical correspondence analysis (CCA) analysis further revealed that the main factors affecting the methanogens community composition were EC (electric conductivity) and pH, and the main factors affecting SRB community composition were pH, SOC and TN, suggesting that pH is a common factor influencing the community compositions of both methanogen and SRB in the sediments of brackish marshes.

High yield in rice mainly depends on large grain weight, ideal plant architecture and proper flowering time adapting to various geographic regions. To help achieve higher yield, phenotype variations of heading date (HD), plant architecture and grain shape in a panel of 416 rice accessions were investigated in this study. A total of 143 markers including 100 simple sequence repeat (SSR) markers and 43 gene-tagged markers were employed in association mapping to detect quantitative trait loci (QTL) responsible for these variations. Among the 7 subpopulations, POP5 in japonica group showed the largest values of HD and grain width (GW), but the smallest values of grain length (GL) and grain length to width ratio (GLW). Among the six indica groups, POP7 had the largest values of HD, GL, GLW, and 1 000-grain weight (TGW). A total of 27 QTLs were detected underlying these phenotypic variations in single year, while 12 of them could be detected in 2006 and 2007. GS3 marker was closely associated with GL, GW and GLW, and widely distributed in different groups. The starch synthesis related gene markers, SSI, SSIIa, SBE1, AGPL4, and ISA1, were linked to plant height (PH), panicle length (PL), flag leaf length (FLL), GW, and GLW. The SSR markers, RM267, RM340 and RM346, were linked to at least two traits. Therefore, these new markers will probably be used to improve rice grain yield or plant architecture when performing marker-assisted selection of proper alleles.

Invasion of plants in wetland ecosystems is often associated with changes in litter decomposition and in nutrient use, uptake and cycling between invasive and native plants. We studied litter decomposition rates, N and P release and elemental composition and stoichiometry during the invasion of Phragmites australis and Spartina alterniflora into native Cyperus malaccensis wetlands in the Minjiang River estuary (China). Aboveground litter in mono-specific stands decomposed faster for Cyperus malaccensis than for Spartina alterniflora and for Phragmites australis. Cyperus malaccensis litter decomposed slower under the stands of both invasive species. In contrast, the litter of both invasive species decomposed faster under Cyperus malaccesis stands. We observed that the invasion of these species was associated with an increased rate of aboveground litter decomposition and large absolute amounts of C, N and P released from the litter when litter from invasive species was mixed with that of native species. Our results suggest that the large nutrient release from litter during early stages of the invasion favored invasive species with larger size and higher nutrient-uptake capacity than the native species.

The removal of phosphorus (P) from fields in the harvested seeds of staple crops is a major driver of the global P cycle, and consequently, there is interest in breeding grain crops with lower seed P concentrations. While it is expected that a reduction in seed total P would result in lower concentrations of the P-rich anti-nutrient phytate, the potential consequences for starch lysophospholipids (LPLs), which affect rice grain quality and health outcomes in humans, are unknown. We examined the accumulation of phytate and starch LPLs in developing indica (cv. IR64) and japonica (cv. Nipponbare) rice grains as well as the stability of starch LPLs and phytate under varying P supply treatments in hydroponics. Source-sink relationships during grain filling (drought and floret abortion) were also investigated in a pot study. Accumulation of phytate and starch LPLs in seeds continued until 18 days after anthesis (DAA) in cv. Nipponbare and 21 DAA in cv. IR64, which mirrored the accumulation of biomass in seeds. In the hydroponic study, permanent withdrawal of P from the nutrient solution at anthesis (until maturity) led to significant reductions in both phytate and starch LPLs with a similar trend observed when P was withdrawn at 8 DAA. In the pot study using soil, alteration of source-sink systems through drought stress during grain filling or floret abortion (manual removal of the top half of each panicle) also led to significant reductions in grain phytate and starch LPLs compared to control plants. These results indicate that in addition to affecting phytate concentrations, reduced P supply to rice plants during grain filling and alteration of source-sink relationships also impacts starch LPL concentrations. Given these complexities, breeding and selection for reduced seed P concentration in rice and other cereals should be undertaken with caution.

Methane production is influenced by the abundance of methanogens and the availability of terminal substrates. Sulfate-reducing bacteria (SRB) also play an important role in the anaerobic decomposition of organic matter. However, the relationships between methane production and methanogen populations, pore water terminal substrates in estuarine brackish marshes are poorly characterized, and even to our knowledge, no published research has explored the relationship between methane production rate and abundance of SRB and pore water dimethyl sulfide (DMS) concentration. We investigated methane production rate, abundances of methanogens and SRB, concentrations of pore water terminal substrates and electron acceptors at a brackish marsh landscape dominated by Phragmites australis, Cyperus malaccensis and Spatina alterniflora marshes zones in the Min River estuary. The average rates of methane production at a soil depth of 30 cm in the three marsh zones were 0.142, 0.058 and 0.067 μg g−1 d−1, respectively. The abundance of both methanogens and SRB in the soil of the P. australis marsh with highest soil organic carbon content was higher than in the C. malaccensis and S. alterniflora marshes. The abundance of methanogens and SRB in the three soil layers was statistically indistinguishable. Mean pore water DMS concentrations at a soil depth of 30 cm under the S. alterniflora marsh were higher than those in the C. malaccensis and P. australis marshes. Methane production rate increased with the abundance of both methanogens and SRB across three marsh zones together at the landscape scale, and also increased with the concentration of pore water acetate, but did not correlate with concentrations of pore water DMS and dissolved CO2. Our results suggest that, provided that substrates are available in ample supply, methanogens can continue to produce methane regardless of whether SRB are prevalent in estuarine brackish marshes.

Projections of climate change impacts over the coming decades suggest that rising sea level will flood coastal wetlands. We studied the impacts of three intensities of flooding on litter decomposition in the native Cyperus malaccensis, and the invasives Spartina alterniflora and Phragmites australis in Shanyutan wetland (Minjiang River estuary, China). Invasive species had larger C, N and P stocks in plant-litter compartments and higher fluxes among plant-litter-soil, which increased with flooding intensity. Litter mass remaining (% of initial mass) were correlated with the N:P ratio in remaining litter, consistently with the N-limitation in this wetland. P. australis had the highest accumulated N release (P

Mineral elements in brown rice grain play an important role in human health. In this study, variations in the content of iron (Fe), zinc (Zn), selenium (Se), cadmium (Cd), and lead (Pb) in 378 accessions of brown rice were investigated, and association mapping was used to detect the quantitative trait loci (QTLs) responsible for the variation. Among seven subpopulations, the mean values of Zn and Cd in the japonica group were significantly higher than in the indica groups. The population structure accounted for from 5.7% (Se) to 22.1% (Pb) of the total variation. Correlation analyses showed that Pb was positively correlated with the other minerals (P < 0.001) except for Se. For the five mineral elements investigated, 20 QTLs, including some previously reported and new candidate loci, were identified. Particularly, three cases of QTL colocalization, i.e. Cd and Pb on chromosome 5, Zn and Pb on chromosome 7, and Se and Pb on chromosome 11, were observed. This study suggested that the identified markers could feasibly be used to enhance desired micronutrients while reducing the heavy metal content in whole rice grain by marker-assisted selection (MAS).

Insulin resistance, a major characteristic of type 2 diabetes (T2D), is closely associated with adipose tissue macrophages (ATMs) that induce chronic low-grade inflammation. Recently, mesenchymal stem cells (MSCs) have been identified in alleviation of insulin resistance. However, the underlying mechanism still remains elusive. Thus, we aimed to investigate whether the effect of MSCs on insulin resistance was related to macrophages phenotypes in adipose tissues of T2D rats. In this study, human umbilical cord-derived MSCs (UC-MSCs) infusion produced significantly anti-diabetic effects and promoted insulin sensitivity in T2D rats that were induced by a high-fat diet combined with streptozotocin and directed ATMs into an alternatively activated phenotype (M2, anti-inflammatory). In vitro, MSC-induced M2 macrophages alleviated insulin resistance caused by classically activated macrophages (M1, pro-inflammatory). Further analysis showed that M1 stimulated UC-MSCs to increase expression of interleukin (IL)-6, a molecule which upregulated IL4R expression, promoted phosphorylation of STAT6 in macrophages, and eventually polarized macrophages into M2 phenotype. Moreover, the UC-MSCs effect on macrophages was largely abrogated by small interfering RNA (siRNA) knockdown of IL-6. Together, our results indicate that UC-MSCs can alleviate insulin resistance in part via production of IL-6 that elicits M2 polarization. Additionally, human obesity and insulin resistance were associated with increased pro-inflammatory ATMs infiltration. Thus, MSCs may be a new treatment for obesity-related insulin resistance and T2D concerning macrophage polarized effects.

Hair follicle morphogenesis and regeneration depend on intensive but well-orchestrated interactions between epithelial and mesenchymal components. Therefore, an alternative strategy to reproduce the process of epithelial-mesenchymal interaction in vitro could use a 3D system containing appropriate cell populations. The 3D air-liquid culture system for reproducibly generating hair follicles from dissociated epithelial and dermal papilla (DP) cells combined with a collagen-chitosan scaffold is described in this study. Wnt-CM was prepared from the supernatant of Wnt1a-expressing bone marrow mesenchymal stem cells (BM-MSCs) that maintain the hair-inducing gene expression of DP cells. The collagen-chitosan scaffold cells (CCS cells) were constructed using a two-step method by inoculating the Wnt-CM-treated DP cells and epidermal (EP) cells into the CCS. The cells in the air-liquid culture formed dermal condensates and a proliferative cell layer in vitro. The CCS cells were able to induce hair regeneration in nude mice. The results demonstrate that Wnt-CM can maintain the hair induction ability of DP cells in expansion cultures, and this approach can be used for large-scale preparation of CCS cells in vitro to treat hair loss. Copyright © 2015 John Wiley & Sons, Ltd.

Nine units in new-born intertidal zone of the Yellow River estuary, China were examined for concentrations of heavy metals (Pb, Cr, Cu, Zn and Ni) in sediments and plants. Heavy metal levels in surface sediments were in the order of Zn > Pb approximate to Cr > Cu approximate to Ni and generally increased in a seaward direction except for Z6 (Tamarix chinensis-Suaeda salsa zone) and Z7 (S. salsa-T. chinensis zone) units. Significant differences in metal concentrations of the 9 units were observed in the profiles (p Cu > Pb > Ni > Cr and differed among plants or tissues. The roots at Z2 (Calamagrostis pseudophragmites zone), Z3 (Imperata cylindrical zone) and Z4 (Phramites australis zone) units accumulated greater metals than shoots [TFs (translocation factors) 1), implying that intertidal plants showed different pathways in metal accumulation and internal transportation. Except for Pb, the concentrations of Cr, Cu, Zn and Ni in sediments were lower than the criteria of Class I recommended by the Environmental Quality Standard for Soils of China, Although heavy metal levels in intertidal zone were generally the lowest (Cr, Cu, Zn and Ni) or relatively moderate (Pb) compared with other estuaries or bays in Asia and Europe, high eco-toxic risk of Pb and Ni exposure still could be observed at Z4, Z6 and Z9 (mudflat zone) units. S. salsa was more suitable for the potential biomonitor or phytoremediation of all five heavy metals if intertidal sediments was seriously contaminated with increasing of pollutants loading in the Yellow River estuary. (C) 2014 Elsevier B.V. All rights reserved.

Human umbilical cord–derived mesenchymal stem cells (UC-MSCs) are multipotent, primitive, and have been widely used for skin tissue engineering. Their transdifferentiation is determined by the local microenvironment. In this study, we investigated the potential epidermal differentiation of UC-MSCs and the formation of epidermis substitutes in a 3-dimensional (3D) microenvironment, which was fabricated by UC-MSCs embedded into collagen–chitosan scaffolds (CCSs) combined with an air–liquid interface (ALI) culture system. Using fluorescence microscope, we observed that UC-MSCs were spindle-shaped and evenly distributed in the scaffold. Methyl thiazolyl blue tetrazolium bromide assay and Live/Dead assay indicated that the CCSs have good biocompatibility with UC-MSCs. Immunohistochemistry and western blotting assay showed that UC-MSCs on the surface of the CCSs were positive for the epidermal markers cytokeratin 19 and involucrin at 14 days. In addition, hematoxylin–eosin staining indicated that multilayered epidermis substitutes were established. The constructed epidermis substitutes were applied to treat full-thickness wounds in rats and proved to promote wound healing. In conclusion, manipulating the 3D microenvironment is a novel method for inducing the epidermal differentiation of MSCs to engineer epidermal substitutes, which provides an alternative strategy for skin tissue engineering.

It is important to establish an efficient vascularization for the long-term acceptance of bioengineered skin equivalents treating the cutaneous wounds of diabetic rats with hindlimb ischemia. This study investigates the possible use of a collagen-chitosan sponge scaffold encapsulated with thymosin beta 4 (CCSS-eTβ4), an angiogenic factor, to accelerate cutaneous wound healing in streptozotocin (STZ)-induced diabetic rats with hindlimb ischemia. CCSSs-eTβ4 was fabricated using a freeze-drying method. The scaffolds were analyzed by scanning electron microscopy, swelling and degradation assays, mechanical properties, and scaffolds of 50:50 collagen-chitosan were selected and applied. The controlled release of Tβ4 from the scaffolds elicited localized and prolonged effects over 12 days, as shown by an enzyme-linked immunosorbent assay (ELISA). In vivo, CCSSs-eTβ4 improved diabetic cutaneous wound healing, with faster wound reepithelialization, better dermal reorganization, and higher wound vascularization. Furthermore, CCSSs-eTβ4 downregulated inflammatory genes and upregulated angiogenic genes in the wound tissue. Significant increases in CD31-positive endothelial cells and new vessel density were also observed. In vitro, Tβ4 increased the migratory and proliferative activity of high glucose (HG)-treated human umbilical vein endothelial cells (HUVECs). Meanwhile, we found that Tβ4 could promote HG-treated HUVECs migration and improve angiogenesis by activation of the VEGF/AKT pathway. Overall, these findings demonstrated the promising potential of CCSSs-eTβ4 to promote more effective wound healing and suggest its possible application for diabetic cutaneous wound treatment.

Methane production is influenced by the abundance of methanogens and the availability of substrates. Sulfate-reducing bacteria (SRB) also play an important role in the anaerobic decomposition of organic matter. However, the relationships between methane production and abundance of methanogen and pore water substrates in estuarine brackish marshes are poorly characterized, and even to our knowledge, no published research has explored the relationship between methane production rate and pore water dimethyl sulfide (DMS) concentration. We investigated methane production rate, abundances of methanogens and SRB, sediment organic carbon contents, and concentrations of pore water substrates (acetate and DMS) and terminal electron acceptors (SO4 2−, NO3 −, and Fe3+) at a brackish marsh landscape dominated by Phragmites australis marsh, Cyperus malaccensis marsh, and Spartina alterniflora marsh in the Min River estuary, southeast China. The average rates of methane production over the entire 30 cm of the sediment profile (5 cm sampling interval) in the three marshes were 0.142, 0.058, and 0.067 μg g−1 day−1, respectively. The abundance of both methanogens and SRB in the sediment of the P. australis marsh with the highest sediment organic carbon content was higher than in the C. malaccensis and S. alterniflora marshes. Mean pore water DMS concentrations over the entire 30 cm of the sediment profile under the S. alterniflora marsh were higher than those in the C. malaccensis marsh and P. australis marsh. Methane production rate correlated weakly with the abundance of methanogens across the three marsh zones together, but did not correlate with the concentrations of pore water acetate and DMS. Our results suggest that the abundance of methanogens is controlled by sediment organic carbon supply, and further, methane production is affected by the abundance of methanogens in the subtropical estuarine brackish marshes.

As a fundamental component of nucleic acids, phospholipids, and adenosine triphosphate, phosphorus (P) is critical to all life forms, however, the molecular mechanism of P translocation and distribution in rice grains are still not understood. Here, with the use of five different low phytic acid (lpa) rice mutants, the redistribution in the main P-containing compounds in rice grain, phytic acid (PA), lysophospholipid (LPL), and inorganic P (Pi), was investigated. The lpa mutants showed a significant decrease in PA and phytate-phosphorus (PA-P) concentration with a concomitant increase in Pi concentration. Moreover, defects in the OsST and OsMIK genes result in a great reduction of specific LPL components and LPL-phosphorus (LPL-P) contents in rice grain. In contrast, defective OsMRP5 and Os2-PGK genes led to a significant increase in individual LPL components. The effect of the Os2-PGK gene on the LPL accumulation was validated using breeding lines derived from a cross between KBNT-lpa (Os2-PGK mutation) and Jiahe218. This study demonstrates that these rice lpa mutants lead to the redistribution of Pi in endosperm and modify LPL biosynthesis. Increase LPLs in the endosperm in the lpa mutants may have practical applications in rice breeding to produce "healthier" rice.

Human ageing is a complex trait that involves the synergistic action of numerous biological processes that interact to form a complex network. Here we performed a network analysis to examine the interrelationships between physiological and psychological functions, disease, disability, quality of life, lifestyle and behavioural risk factors for ageing in a cohort of 3,270 subjects aged ≥55 years. We considered associations between numerical and categorical descriptors using effect-size measures for each variable pair and identified clusters of variables from the resulting pairwise effect-size network and minimum spanning tree. We show, by way of a correspondence analysis between the two sets of clusters, that they correspond to coarse-grained and fine-grained structure of the network relationships. The clusters obtained from the minimum spanning tree mapped to various conceptual domains and corresponded to physiological and syndromic states. Hierarchical ordering of these clusters identified six common themes based on interactions with physiological systems and common underlying substrates of age-associated morbidity and disease chronicity, functional disability, and quality of life. These findings provide a starting point for indepth analyses of ageing that incorporate immunologic, metabolomic and proteomic biomarkers, and ultimately offer low-level-based typologies of healthy and unhealthy ageing. ASTAR (Agency for Sci., Tech. and Research, S’pore) Published version