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Abstract Introduction As a crucial component of the nitrogen cycle, ammonia oxidation in soil can be driven by canonical ammonia‐oxidising archaea (AOA) and bacteria, as well as complete ammonia oxidiser (CMX Nitrospira). Land use change can disrupt and alter the soil microbial community and the nitrogen cycle. Materials & Methods We compared the soil ammonia‐oxidising microorganisms and ammonia oxidation potentials in a coastal marsh and nearby reclaimed aquaculture ponds, monthly over a 10‐month period in southeastern China. The abundance of ammonia oxidisers was assessed by real‐time quantitative PCR and the community structure of CMX Nitrospira was evaluated by high‐throughput sequencing. Results The ammonia oxidiser community was dominated by AOA in the marsh (91%) and was made up of similar proportions of AOA and CMX Nitrospira in the aquaculture ponds (46%–47%). The CMX Nitrospira community structure changed significantly between habitat types, mainly driven by opposite change in relative abundance of clade B versus clades A.2 and A.3. Aquaculture reclamation decreased the soil potential ammonia oxidation rate (PAO) by an order of magnitude, and AOA was the only significant predictor of PAO among all ammonia oxidiser groups. Conclusion Our results suggest that aquaculture reclamation from coastal marshes would significantly alter the soil ammonia oxidiser community and decrease ammonia oxidation rate, and CMX Nitrospira appear to play a relative larger role in nitrogen cycling in aquaculture ponds.

Abstract Although chimeric antigen receptor (CAR) T cells have become an important treatment option for patients with relapsed/refractory B-cell malignancies, more than 60% of patients with diffuse large B-cell lymphoma (DLBCL) treated with CAR-T cell therapies fail to achieve a durable response. To reveal changes in CAR-T cell therapy and identify response biomarkers, we conducted a retrospective analysis of pre-manufacture source T cells and CAR-T cell products and their association with outcome in 58 patients with r/rDLBCL who received tandem CD19/CD20 CAR-T cell therapy. We performed bulk RNA-Seq, single-cell RNA-Seq, and paired T cell receptor sequencing on CAR-T cell products and pre-manufacture T cells from DLBCL patients. We note that a CD8+ stem cell-like memory T cell population with a higher proportion and enhanced activating capacity of the CAR-T cell products was key to achieving durable clinical response. By analysing autologously-derived, pre-manufacture T cells, our data suggest that heterogeneity in the cellular and molecular features of pre-manufacture T cells contribute to the variation in efficacy after CAR-T cell therapy in DLBCL. The differences in anti-tumour efficacy of CAR-T cells among patients with different clinical outcomes appear to be due to the loss of CCR7 gene expression, coupled with increased expression of activation- and inhibitor-related genes in the CD8+ naïve-T cell populations among the apheresis T cells from patients with a poor molecular response. These findings significantly advance our understanding of the underlying molecular determinants of pre-manufacture T cell function.

Abstract Starch is the most important component in potato tubers, whose structure and composition play key roles in properties of potato storage, processing, and applications. This review identifies the biodiversity of starch content, structure, and physiochemical properties, particularly the amylose and amylopectin content, molecular and granular structure, gelatinization, retrogradation, swelling power, solubility, and digestibility of potato in the past decade. Various functions of major potato starch biosynthesis‐related enzymes, such as AGPase, starch synthase, starch branching enzyme, debranching enzyme, and glucan/water dikinase, are also summarized. The potential utilization of native and modified potato starch in food and medicine fields owing to its high pasting viscosity is also analyzed. This review will contribute to a better understanding of the genetics–structure–functions of potato starch and may provide insights into potential potato starch utilization and further genetic improvement of starch quality.

IntroductionTubulin epsilon and delta complex 2 (TEDC2) is widely expressed in various human tissues and primarily governs centriole stability. However, the biological significance of TEDC2 in pan-cancer is unclear.MethodsIn this study, we employed R software and various online bioinformatics analysis tools to investigate the functional attributes of TEDC2 in human tumours and its potential involvement in immune response. The status of TEDC2 expression was evaluated in samples from the TCGA and GEO datasets, as well as in tumour and corresponding normal samples from the TCGA database. Subsequently, Kaplan-Meier estimates, clinical correlations, and univariate Cox regressions were used to analyze the 33 types of tumors from TCGA and determine the prognostic significance of TEDC2. Moreover, nomogram models were formulated using three distinct tumours, namely kidney renal clear cell carcinoma (KIRC), lung adenocarcinoma (LUAD), and liver hepatocellular carcinoma (LIHC), to evaluate the prognostic significance of TEDC2 in tumours. Furthermore, TEDC2 was investigated for its correlation with the levels of immune cell infiltration, and a functional enrichment analysis was conducted to identify potential signalling pathways involving TEDC2. ResultsDifferential analysis revealed that 16 tumour types expressed TEDC2 to a greater extent than normal tissues. The abnormal expression of TEDC2 can predict survival outcomes in patients with adrenocortical carcinoma (ACC), KIRC, kidney renal papillary cell carcinoma (KIRP), LUAD, LIHC, lower grade glioma (LGG), and thymoma (THYM). Subsequent results indicated that TEDC2 has the ability to influence ECM regulators, cell cycle, and Immune checkpoint-associated signalling pathways, which could potentially lead to a poor prognosis and tumour progression.DiscussionTEDC2 has been identified as a potential therapeutic target that could predict the prognosis of multiple tumour types, making it a promising target for reversing tumour development.

Abstract Despite the remarkable success of chimeric antigen receptor (CAR) T-cell therapy for treating hematologic malignancies, resistance and recurrence still occur, while the markers or mechanisms underlying this resistance remain poorly understood. Here, via an unbiased genome-wide CRISPR/Cas9 screening, we identified loss of NOXA, a B-cell lymphoma 2 (BCL2) family protein in B-cell malignancies, as a pivotal regulator of resistance to CAR T-cell therapy by impairing apoptosis of tumor cells both in vitro and in vivo. Notably, low NOXA expression in tumor samples was correlated with worse survival in a tandem CD19/20 CAR T clinical trial in relapsed/refractory B-cell lymphoma. In contrast, pharmacological augmentation of NOXA expression by histone deacetylase (HDAC) inhibitors dramatically sensitized cancer cells to CAR T cell-mediated clearance in vitro and in vivo. Our work revealed the essentiality of NOXA in resistance to CAR T-cell therapy and suggested NOXA as a predictive marker for response and survival in patients receiving CAR T-cell transfusions. Pharmacological targeting of NOXA might provide an innovative therapeutic strategy to enhance CAR T-cell therapy.

Microbial evolution-mediated CO2 from litter has aroused widespread concern, and knowing the factors controlling litter-derived CO2 is important when considering the effects of accumulative CO2 release from litter on the global greenhouse. We conducted a short-term N addition (6, 16, and 24 g N m‒2 yr‒1) experiment in Cyperus malaccensis var. brevifolius (shichito matgrass) litter decomosition. Phospholipid fatty acid (PLFA) method and enzyme method were used to analysis litter microbial community composition and enzymatic activity. During a 220-day decomposition period, there was little effect of the N amendments on litter CO2 evolution rates (9.97‒307.54 μg C g−1 h−1) with a notable exception regarding the increase of the high-N treatment at day 20. The accumulative CO2 release significantly increased after N addition in the medium and late phases. The facilitation effect on accumulative CO2 release by N amendments was more and more obvious over the decomposition time, especially for the low- and intermediate-N treatments. At the end of our experiment, compared with the control treatment, accumulative CO2 release increased 69.75%, 76.62%, and 39.93% for low-, intermediate-, and high-N treatments, respectively. These observations highlight that N deposition could cause high losses of litter C as CO2.

Abstract 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.

Sweet potato flour is a key ingredient for the production of new food products worldwide, which imparts desired properties, nutritional value, antioxidants, and natural color to processed foods. However, little information regarding the functional properties of the sweet potato flour is available. In this study, the genetic diversity in the physiochemical, nutritional, and antioxidant properties of wholemeal flour from seven sweet potato varieties was investigated. The total phenolic content (TPC) of the free and bound fractions ranged from 13.85 to 90.74 mg gallic acid equivalent (GAE)/100 g and from 5.07 to 24.29 mg GAE/100 g, respectively. The average protein content of sweet potato was 5.41 g/100 g ranging from 3.40 to 8.60 g/100 g DW. The total amino acid content averaged 45.13 mg/g DW. The average contents of 12 mineral elements were in the order of K > P > Ca > Mg > Mn > Fe > Zn > Cu > Ni > Se > Cr > Cd. K and P contents were the highest among all accessions, which were positively correlated with most of the other minerals. The average starch content of sweet potato was 53.90 g/100 g DW, ranging from 31.68 to 64.90%. The peak viscosity (PV), hot paste viscosity (HPV), and cold paste viscosity (CPV) were in the range of 90.7–318.8 Rapid Visco Unit (RVU), 77.3–208.3 RVU, and 102.6–272.7 RVU, respectively. The hardness values and cohesiveness (Coh) varied among different sweet potatoes, with a range of 8.20–18.48 g and 0.22–0.68, respectively. The gelatinization onset, peak, conclusion temperatures, and enthalpy were in the ranges of 59.39–71.91°C, 70.19–88.40°C, 78.98–95.79°C, 1.85–5.65 J/g, respectively.

De novo DNA methylation has been associated with T cell exhaustion in cancer immunotherapy. Here the authors show that the pre-treatment of CD19 CAR-T cells with the DNA methyltransferase inhibitor decitabine limits exhaustion and confers enhanced proliferative, effector and memory properties upon antigen exposure, with improved tumor control.

Abstract 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 .

Abstract Background Blocking programmed death-1 (PD-1) is considered to be a promising strategy to improve T cell function, and this is being explored in many ongoing clinical trials. In fact, our knowledge about PD-1 is primarily based on the results of short-term experiments or observations, but how long-lasting PD-1 blockade can affect T cell function remains unclear. Methods We planned to use shRNA-based gene knockdown technology to mimic long-lasting PD-1 blockade. We constructed PD-1 steadily blocked chimeric antigen receptor modified T (CAR-T) cells, and with these cells we can clearly study the effects of PD-1 knockdown on T cell function. The anti-tumor function, proliferation ability and differentiation status of PD-1 silenced CAR-T cells were studied by in vitro and animal experiments. Results According to short-term in vitro results, it was reconfirmed that the resistance to programmed death-ligand 1 (PD-L1)-mediated immunosuppression could be enhanced by PD-1 blockade. However, better anti-tumor function was not presented by PD-1 blocked CAR-T cells in vitro or in vivo experiments. It was found that PD-1 knockdownmight impair the anti-tumor potential of CAR-T cells because it inhibited T cells’ proliferation activity. In addition, we observed that PD-1 blockade would accelerate T cells’ early differentiation and prevent effector T cells from differentiating into effect memory T cells, and this might be the reason for the limited proliferation of PD-1 silenced CAR-T cells. Conclusion These results suggest that PD-1 might play an important role in maintaining the proper proliferation and differentiation of T cells, and PD-1 silencing would impair T cells’ anti-tumor function by inhibiting their proliferation activity.

Abstract 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.

Expressed by cancer stem cells of various epithelial cell origins and hepatocellular carcinoma (HCC), CD133 is an attractive therapeutic target for HCC. The marker CD133 is highly expressed in endothelial progenitor cells (EPC). EPCs circulate in increased numbers in the peripheral blood of patients with highly vascularized HCC and contribute to angiogenesis and neovascularization. This phase II study investigated CD133-directed chimeric antigen receptor (CAR) T (CART-133) cells in adults with HCC. Patients with histologically confirmed and measurable advanced HCC and adequate hematologic, hepatic, and renal functions received CART-133 cell infusions. The primary endpoints were safety in phase I and progression-free survival (PFS) and overall survival (OS) in phase II. Other endpoints included biomarkers for CART-133 T cell therapy. Between June 1, 2015, and September 1, 2017, this study enrolled 21 patients who subsequently received CART-133 T cells across phases I and II. The median OS was 12 months (95% CI, 9.3–15.3 months) and the median PFS was 6.8 months (95% CI, 4.3–8.4 months). Of 21 evaluable patients, 1 had a partial response, 14 had stable disease for 2 to 16.3 months, and 6 progressed after T-cell infusion. The most common high-grade adverse event was hyperbilirubinemia. Outcome was correlated with the baseline levels of vascular endothelial growth factor (VEGF), soluble VEGF receptor 2 (sVEGFR2), stromal cell-derived factor (SDF)-1, and EPC counts. Changes in EPC counts, VEGF, SDF-1, sVEGFR2, and interferon (IFN)-γ after cell infusion were associated with survival. In patients with previously treated advanced HCC, CART-133 cell therapy demonstrates promising antitumor activity and a manageable safety profile. We identified early changes in circulating molecules as potential biomarkers of response to CART-133 cells. The predictive value of these proangiogenic and inflammatory factors as potential biomarkers of CART-133 cell therapy in HCC will be explored in prospective trials. This study is registered at ClinicalTrials.gov (NCT02541370)

Abstract 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.

Silica (Si) is a basic nutrient requirement for many aquatic organisms and its biogeochemical cycle plays an important role in estuarine coastal ecosystems. However, little is known about the role Si plays during plant–plant interactive processes in the marsh ecosystems. Here, variations in biomass, biogenic silica (BSi) content, and available Si content of Cyperus malaccensis-dominated marshes, Phragmites australis-dominated marshes, and their ecotonal marshes were studied in the Shanyutan marsh in the Min River estuary, China. Results showed that C. malaccensis and P. australis biomass in ecotones was lower than those in typical communities by 46.4% and 46.3%, respectively. BSi content in aboveground organs of C. malaccensis and culms and roots of P. australis was lower in ecotones than in typical communities, whereas BSi content in other organs showed the opposite trend. Biomass allocation in C. malaccensis and P. australis roots in ecotones was higher by 56.9% and 19.5%, respectively, and BSi stock in C. malaccensis and P. australis roots was higher than that in typical communities by 120.9% and 18.9%, respectively. Available Si content in ecotonal marsh soils was 12.6% greater than that in typical communities. Thus, the two plant species may use different strategies for Si accumulation and allocation in ecotones to adapt to the competitive environment. P. australis may expand primarily via occupation of wider aboveground space, thereby increasing the Si accumulation capacity in aboveground organs. Meanwhile, C. malaccensis may increase the Si allocation capacity of its roots to withstand the pressure from P. australis. This study will provide new insights into marsh plant competition from the perspective of Si, which can also benefit plant management in marsh ecosystems.

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.

Table of contents A1 Proceedings of 2016 China Cancer Immunotherapy Workshop, Beijing, China Bin Xue, Jiaqi Xu, Wenru Song, Zhimin Yang, Ke Liu, Zihai Li A2 Set the stage: fundamental immunology in forty minutes Zihai Li A3 What have we learnt from the anti-PD-1/PD-L1 therapy of advanced human cancer? Lieping Chen A4 Immune checkpoint inhibitors in lung cancer Edward B. Garon A5 Mechanisms of response and resistance to checkpoint inhibitors in melanoma Siwen Hu-Lieskovan A6 Checkpoint inhibitor immunotherapy in lymphoid malignancies Wei Ding A7 Translational research to improve the efficacy of immunotherapy in genitourinary malignancies Chong-Xian Pan A8 Immune checkpoint inhibitors in gastrointestinal malignancies Weijing Sun A9 What’s next beyond PD-1/PDL1? Yong-Jun Liu A10 Cancer vaccines: new insights into the oldest immunotherapy strategy Lei Zheng A11 Bispecific antibodies for cancer immunotherapy Delong Liu A12 Updates on CAR-T immunotherapy Michel Sadelain A13 Adoptive T cell therapy: personalizing cancer treatment Cassian Yee A14 Immune targets and neoantigens for cancer immunotherapy Rongfu Wang A15 Phase I/IIa trial of chimeric antigen receptor modified T cells against CD133 in patients with advanced and metastatic solid tumors Meixia Chen, Yao Wang, Zhiqiang Wu, Hanren Dai, Can Luo, Yang Liu, Chuan Tong, Yelei Guo, Qingming Yang, Weidong Han A16 Cancer immunotherapy biomarkers: progress and issues Lisa H. Butterfield A17 Shaping of immunotherapy response by cancer genomes Timothy A. Chan A18 Unique development consideration for cancer immunotherapy Wenru Song A19 Immunotherapy combination Ruirong Yuan A20 Immunotherapy combination with radiotherapy Bo Lu A21 Cancer immunotherapy: past, present and future Ke Liu A22 Breakthrough therapy designation drug development and approval Max Ning A23 Current European regulation of innovative oncology medicines: opportunities for immunotherapy Harald Enzmann, Heinz Zwierzina

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Expressed by cancer stem cells of various epithelial cell origins, CD133 is an attractive therapeutic target for cancers. Autologous chimeric antigen receptor-modified T-cell directed CD133 (CART-133) was first tested in this trial. The anti-tumor specificity and the postulated toxicities of CART-133 were first assessed. Then, we conducted a phase I clinical study in which patients with advanced and CD133-positive tumors received CART-133 cell-infusion. We enrolled 23 patients (14 with hepatocellular carcinoma [HCC], 7 with pancreatic carcinomas, and 2 with colorectal carcinomas). The 8 initially enrolled patients with HCC were treated by a CART-133 cell dose escalation scheme (0.05–2 × 106/kg). The higher CAR-copy numbers and its reverse relationship with the count of CD133+ cells in peripheral blood led to the determination of an acceptable cell dose is 0.5–2 × 106/kg and reinfusion cycle in 23 patients. The primary toxicity is a decrease in hemoglobin/platelet (≤ grade 3) that is self-recovered within 1 week. Of 23 patients, three achieved partial remission, and 14 achieved stable disease. The 3-month disease control rate was 65.2%, and the median progression-free survival was 5 months. Repeated cell infusions seemed to provide a longer period of disease stability, especially in patients who achieved tumor reduction after the first cell-infusion. 21 out of 23 patients had not developed detectable de novo lesions during this term. Analysis of biopsied tissues by immunohistochemistry showed CD133+ cells were eliminated after CART-133 infusions. This trial showed the feasibility, controllable toxicities, and effective activity of CART-133 transfer for treating patients with CD133-postive and late-stage metastasis malignancies.

Mesenchymal stem cells (MSCs) can affect the microenvironment of a wound and thereby accelerate wound healing. Wnt proteins act as key mediators of skin development and participate in the formation of skin appendages such as hair. The mechanisms of action of MSCs and Wnt proteins on skin wounds are largely unknown. Here, we prepared a Wnt7a-containing conditioned medium (Wnt-CM) from the supernatant of cultured human umbilical cord-MSCs (UC-MSCs) overexpressing Wnt7a in order to examine the effects of this CM on cutaneous healing. Our results revealed that Wnt-CM can accelerate wound closure and induce regeneration of hair follicles. Meanwhile, Wnt-CM enhanced expression of extracellular matrix (ECM) components and cell migration of fibroblasts but inhibited the migratory ability and expression of K6 and K16 in keratinocytes by enhancing expression of c-Myc. However, we found that the CM of fibroblasts treated with Wnt-CM (HFWnt-CM-CM) can also promote wound repair and keratinocyte migration; but there was no increase in the number of hair follicles of regeneration. These data indicate that Wnt7a and UC-MSCs have synergistic effects: they can accelerate wound repair and induce hair regeneration via cellular communication in the wound microenvironment. Thus, this study opens up new avenues of research on the mechanisms underlying wound repair.

Understanding the functional relationship between greenhouse gas fluxes and environmental variables is crucial for predicting the impacts of wetlands on future climate change in response to various perturbations. We examined the relationships between methane (CH4) emission and temperature in two marsh stands dominated by the Phragmites australis and Cyperus malaccensis, respectively, in a subtropical estuarine wetland in southeast China based on three years of measurement data (2007-2009). We found that the Q10 coefficient of CH4 emission to soil temperature (Qs10) from the two marsh stands varied slightly over the three years (P > 0.05), with a mean value of 3.38 ± 0.46 and 3.89 ± 0.41 for the P. australis and C. malaccensis stands, respectively. On the other hand, the three-year mean Qa10 values (Q10 coefficients of CH4 emission to air temperature) were 3.39 ± 0.59 and 4.68 ± 1.10 for the P. australis and C. malaccensis stands, respectively, with a significantly higher Qa10 value for the C. malaccensis stand in 2008 (P < 0.05). The seasonal variations of Q10 (Qs10 and Qa10) differed among years, with generally higher values in the cold months than those in the warm months in 2007 and 2009. We found that the Qs10 values of both stands were negatively correlated with soil conductivity, but did not obtain any conclusive results about the difference in Q10 of CH4 emission between the two tidal stages (before flooding and after ebbing). There were no significant differences in both Qs10 and Qa10 values of CH4 emission between the P. australis stand and the C. malaccensis stands (P > 0.05). Our results show that the Q10 values of CH4 emission in this estuarine marsh are highly variable across space and time. Given that the overall CH4 flux is governed by a suite of environmental factors, the Q10 values derived from field measurements should only be considered as a semi-empirical parameter for simulating CH4 emissions.

Endothelial dysfunction induced by unordered metabolism results in vascular reconstruction challenges in diabetic lower limb ischemia (DLLI). Mesenchymal stem cells (MSCs) are multipotent secretory cells that are suitable for clinical DLLI treatment, but their use has been hampered by poor survival after injection. Hypoxia can significantly enhance the capacity of MSCs to secrete angiogenic factors. We investigated transient hypoxia pretreatment of MSCs to facilitate revascularization in DLLI. Rat bone marrow MSCs (BM-MSCs) were cultured at different oxygen concentrations for varying time periods. The results indicated that transient pretreatment (5% O2, 48 h) not only increased the expression of VEGF-1α, ANG, HIF-1α and MMP-9 in BM-MSCs as assessed by real-time RT-PCR, but also increased the expression of Bcl-2 as determined by western blotting. The transplantation of pretreated BM-MSCs into rats with DLLI demonstrated accelerated vascular reconstruction when assayed by angiography and immunohistochemistry. CM-Dil-labeled tracer experiments indicated that the survival of BM-MSCs was significantly improved, with approximately 5% of the injected cells remaining alive at 14 days. The expression levels of VEGF-1α, MMP-9 and VEGF-R were significantly increased, and the expression of pAKT was up-regulated in ischemic muscle. Double immunofluorescence studies confirmed that the pretreated BM-MSCs promoted the proliferation and inhibited the apoptosis of endothelial cells. In vitro, pretreated BM-MSCs increased the migratory and tube forming capacity of endothelial cells (ECs). Hypoxia pretreatment of BM-MSCs significantly improved angiogenesis in response to tissue ischemia by ameliorating endothelial cell dysfunction and is a promising therapeutic treatment for DLLI.

To uncover the genetics of rice grain weight, we constructed an RIL population derived from a cross between a large grain accession M201 and a small size variety JY293. Specific Locus Amplified Fragment Sequencing (SLAF-Seq) technology was used to genotype two bulked DNA pools made from individual DNA of the heaviest 30 lines and the lightest 30 lines according to the 1000 grain weight (TGW). Bulked segregant analysis (BSA) was used to identify SLAFs strongly associated with TGW. Two marker-intensive regions at 24,600,000-24,850,000 bp and 25,000,000-25,350,000 bp on chromosome 3 were identified tightly related to the TGW. Then a linkage map of chromosome 3 was constructed with SSR markers and some SLAF derived single nucleotide polymorphisms (SNPs). Quantitative trait locus (QTL) mapping for TGW, grain length, grain width, and grain thickness revealed one major QTL in the second hot-region and two other minor QTLs for grain weight. These three QTLs displayed hierarchical effects on grain length and grain weight in order of qTGW3.2 (qGL3) qTGW3.1 (GS3) qTGW3.3. Multiple comparisons of means among the eight combinations of 3 QTLs revealed that the lines with two of three QTLs deriving from M201 displayed a large grain weight phenotype (TGW 40.2g, average data of three years) and lines with both qTGW3.1 and qTGW3.3 alleles from M201 (42.5g) had similar grain weight to the qTGW3.2 (40.8g) alone. Two strategies with similar effectiveness were proposed to improve grain weight by marker-assisted selection (MAS). One is to introduce the novel qTGW3.2 allele alone, and the other is to pyramid qTGW3.1 and qTGW3.3 alleles together. One new allele of GS3 (39 bp deletion in intron 1) and two SNPs in coding sequence of qGL3 identified in this study from M201 are useful in pyramiding elite alleles for molecular breeding for improvement of rice yield.

Litter production and decomposition are key processes controlling the capacity of wetland to store and cycle carbon (C) and nutrients. Typhoons deposit large amounts of green and semi-green (between green and withered) plant tissues and withered litter (normal litter) on wetland soils, generating a pulse of litter production. Climatic models project an increase in typhoon intensity and frequency. Elucidating the impacts of typhoons on C, N and P cycles and storage capacities in subtropical and tropical wetland areas is thus important. We analyzed the patterns and changes of litter decomposition after a typhoon in the Minjiang River estuary in southeastern China. Green litter decomposed the fastest, and the loss of mass did not differ significantly between semi-green litter, withered litter and mixed litter (all soil litter after a typhoon). During the decomposition process the remaining green litter had the highest, and withered litter the lowest N and P concentrations. The biomass loss rate of litter during the studied period was related to the initial litter N and P concentrations. Remaining litter generally increased its N:P ratio during decomposition. The ratio of the released N and P was consequently lower than the initial N:P ratio in all litter types. The typhoon enhanced the release of C, N and P from the litter (884, 12.3 and 6 kg ha ^−1 , respectively) by 264 days after the typhoon. The soil was accordingly enriched with organic matter and nutrients for several months, which should favor microbial growth rates (higher C, N and P availability and lower C:nutrient and N:P ratios) and increase the rates of C and nutrient cycling. If the frequency and/or intensity of typhoons increase, a constant increase in the release of N and P to the soil with lower N:P ratios could change the N and P cycles in wetlands and provide better conditions for the spread of fast-growing species.