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SummaryTo explore whether embryo culture with melatonin (MT) can improve the embryonic development and clinical outcome of patients with repeated cycles after in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) failure, immature oocytes from controlled ovarian superovulation cycles were collected for in vitro maturation (IVM) and ICSI. The obtained embryos were cultured in 0, 10–11, 10–9, 10–7 and 10–5 M MT medium respectively, and 10–9 M was screened out as the optimal concentration. Subsequently, 140 patients who underwent failed IVF/ICSI cycles received 140 cycles of embryo culture in vitro with a medium containing 10–9 M MT, these 140 MT culture cycles were designated as the experimental group (10–9 M group), and the control group was the previous failed cycles of patients (0 M group). The results showed that the fertilization, cleavage, high-quality embryo, blastocyst, and high-quality blastocyst rates of the 10–9 M group were significantly higher than those of the 0 M group (P < 0.01; P < 0.01; P < 0.0001; P < 0.0001; P < 0.0001). To date, in total, 50 vitrified-warmed cycle transfers have been performed in the 10–9 M group and the implantation rate, biochemical pregnancy rate and clinical pregnancy rate were significantly higher than those in the 0 M group (all P < 0.0001). Two healthy infants were delivered successfully and the other 18 women who achieved clinical pregnancy also had good examination indexes. Therefore the application of 10–9 M MT to embryo cultures in vitro improved embryonic development in patients with repeated cycles after failed IVF/ICSI cycles and had good clinical outcomes.Trial registration: ChiCTR2100045552.

STUDY QUESTION Is it possible to establish a new in-vitro activation (IVA) protocol for infertility treatment? SUMMARY ANSWER A new IVA procedure is an efficient and easily performed approach for infertility treatment of patients with diminished ovarian reserve (DOR). WHAT IS KNOWN ALREADY IVA of primordial follicles with or without stimulators has been developed to treat patients with primary ovarian insufficiency (POI) successfully. However, the efficiency of the procedure is still very low. There is a requirement to optimize the protocol with increased efficiency for clinical application. STUDY DESIGN, SIZE, DURATION Newborn mouse ovaries were used to establish a new 1-h IVA protocol with the mechanistic target of rapamycin (mTOR) stimulator phosphatidic acid (PA, 200 µM) and the phosphatidylinositol-3-kinase (PI3K) stimulator 740Y-P (250 µg/ml); a prospective observational cohort study in POI patients was performed on 15 POI patients and 3 poor ovarian response (POR) patients in three different centers of reproductive medicine in China. PARTICIPANTS/MATERIALS, SETTING, METHODS One-third of ovarian cortex was removed and processed into bigger strips (1 × 1 cm2, 1–2 mm thickness). Strips were then sutured back after treatment. The new approach only requires one laparoscopic surgery. MAIN RESULTS AND THE ROLE OF CHANCE Follicular activation and development increased in cultured mouse and human ovarian tissues after 1 h of stimulator treatment. Compared with tiny ovarian cortex pieces (1 × 1 mm2), large ovarian strips (1 × 1 cm2) showed the lowest apoptotic signals after incubation. We applied the orthotropic transplantation procedure with large strips in the clinic, and 9 of 15 POI patients showed at least one-wave follicular growth during the monitoring period. One patient was reported with one healthy delivery after natural conception and another patient with a healthy singleton delivery after IVF. All the contacted patients (n = 13) responded with no side effects on their health 2–4 years after IVA procedure. LIMITATIONS, REASONS FOR CAUTION Further clinical trials with a large number of well-defined patients are required to compare different IVA protocols. A long-term follow-up system should be set up to monitor patient’s health in the future cohort study. WIDER IMPLICATIONS OF THE FINDINGS By using stimulators, the findings in the study provide a more efficient IVA protocol for the treatment of patients with DOR. It requires only one laparoscopic surgery and thus minimizes patients’ discomfort and costs. This strategy could be useful for patients diagnosed with POI and desire pregnancy as soon as possible after the operation. STUDY FUNDING/COMPETING INTEREST(S) This work was supported by the National Key Research and Development Program of China (2018YFC1003703 and 2018YFC1004203); the National Natural Science Foundation of China (81871221); Co-construction of Provincial Department (201601006). The authors have no conflict of interest to disclose. TRIAL REGISTRATION NUMBER ChiCTR2000030872.

Quantitative differential phase contrast (qDPC) imaging has become an important method of optical measurement and life science research in microscopy because of its high reconstruction resolution and non-invasive, high-contrast and quantitative imaging of biological samples. Despite the continuous development of the principle and algorithm, the frame rate of the existing qDPC algorithm is still much lower than that of camera acquisition, so it is hardly applied to real-time image the fast-moving biological samples. In this paper, based on color-coded multiplexing strategy, a compact real-time quantitative phase imaging system is designed to realize multi-mode imaging. The system employs a programmable LED array to illuminate directly, and the phase reconstruction algorithm is deployed in the graphics processing unit (GPU) of the laptop to accelerate the calculation. The system can achieve high-speed quantitative phase imaging of non-stained biological samples, and the frame rate can reach 60fps. The device has the advantages of compact structure, low cost and portability. Thus, it is suitable for mobile medical applications.

SummaryTo explore whether different polyvinylpyrrolidone (PVP) concentrations affect the results of intracytoplasmic sperm injection (ICSI), a prospective study was conducted for 194 couples undergoing 210 ICSI therapy cycles. These cycles were divided into three groups (10, 7 and 5% groups) using the corresponding concentration of PVP for sperm immobilization. The main outcome measures were analyzed. Results indicated that, with a decrease in PVP concentrations, all of the main outcome measures increased. In particular, the high-quality cleavage embryo rate in the 7% group was significantly lower than in the 5% group (P < 0.01), and the cleavage, high-quality cleavage embryo, and high-quality blastocyst rates in the 5% group were significantly higher than those in the 10% group (all P < 0.001). For high-/intermediate-quality semen, all of the main outcome measures were significantly increased with 5% PVP. For the poor-quality semen, only the high-quality cleavage embryo and high-quality blastocyst rates were significantly higher in the 5% group. Therefore, lowering PVP concentrations greatly promoted the development of embryos in ICSI cycles, with an optimal concentration of 5% for ICSI.

Shunqing Su,1 Xiumei Ding,1 Huijuan Zou,1 Yuechun Lin,1 Jianmin Huang,1 Disheng Xiong,1 Jinan Kuan,1 Yanhong Zhang,2 Rurong Xie2 1Department of Burn&Plast, Dalang Hospital of Dongguan, Dongguang, Guangdong, 523780, People’s Republic of China; 2Department of Wound Repair, Hongmei Hospital of Dongguan, Dongguang, Guangdong, 523160, People’s Republic of ChinaCorrespondence: Xiumei DingDepartment of Burn&Plast, Dalang Hospital of Dongguan, No. 85 of Jinlangzhong Street, Dalang District, Dongguang, 523780, People’s Republic of ChinaTel +86 769-83009512Fax +86 769-83033122Email ding_xm123@outlook.comObjective: The present study aims to explore the individualized treatment options for multisite pressure ulcer (PU) at various stages in elderly patients with multiple medical conditions.Methods: Stages 1 and 2 PU at 146 sites were treated with closed negative pressure suction combined with continuous micro-oxygen perfusion and the local application of foam dressings, silver ion dressings, and moist burn cream. Stages 3 and 4 PU in the sacrococcygeal region were treated with skin or myocutaneous flap transplantation.Results: Stages 1 and 2 PU healed after treatment with closed negative pressure suction combined with continuous micro-oxygen perfusion and dressing changes. One case died during hospitalization due to an illness. Skin or myocutaneous flap repair was conducted in 34 cases of stage 3 or 4 PU in the sacrococcygeal area. Of these cases, 28 achieved primary healing, and 6 required two or three surgeries, 5 of which received micro-skin implantation. In addition, 10 small deep PU at other sites were repaired by direct excision and suturing or local flap repair. Seven cases were transferred to other departments or hospitals due to concomitant diseases or were discharged automatically without surgical treatment.Conclusion: Home care for geriatric patients is difficult. PU often occur at multiple sites because of the duration of various pressures, and different sites may demonstrate different stages because of varying degrees of pressure. When actively treating stages 3 and 4 PU, the trauma management of stages 1 and 2 PU should not be neglected.Keywords: oldest-old patients, pressure ulcer, wound management, flap transplantation, micro-skin implantation

Background: Melatonin (MT), a powerful antioxidant, can effectively ameliorate the in vitro development of animal embryos, but few studies have been performed on human embryos. Therefore, we investigated whether the application of MT in embryo cultures can improve embryonic development and clinical outcomes of patients with repeated cycles after failed in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) cycles.Methods: Human immature oocytes from controlled ovarian hyperstimulation cycles were collected for in vitro maturation culture and ICSI fertilization. The obtained embryos were cultured in vitro in medium containing 0, 10-11, 10-9, 10-7 or 10-5 M MT, and 10-9 M was determined to be the optimal concentration. Subsequently, 140 patients who experienced failed IVF/ICSI cycles underwent 140 cycles of embryo culture in vitro with medium containing 10-9 M MT. High-quality blastocysts were collected and cryopreserved for three months before vitrified-warmed embryo transfer. These culture cycles served as the experimental (10-9 M) group. The control (0 M) group comprised previous failed cycles. The patients were further divided into subgroups of 1, 2 or ≥3 failed cycles. The fertilization and embryo development statuses were compared.Results: The fertilization, cleavage, high-quality embryo, blastocyst, and high-quality blastocyst rates of the 10-9 M group were significantly higher than those of the 0 M group (87.7% vs. 83.6%, p p p -9 M group and the implantation rate, biochemical pregnancy rate and clinical pregnancy rate were significantly higher than those in the 0 M group (65.6% vs. 9.7%, p p p Conclusion: The application of MT to embryo cultures in vitro improved embryonic development in patients with repeated cycles after failed IVF/ICSI cycles and had good clinical outcomes. The optimal concentration of MT was 10-9 M.Trial registration: Name in the registry: Effect of melatonin on the clinical outcome of patients with repeated cycles after failed cycles of in vitro fertilization and intracytoplasmic sperm injection; registration number: ChiCTR2100045552; date of registration: April 19, 2021(retrospectively registered); URL of trial registry record: www.medresman.org.cn.

Background: Melatonin (MT), as a powerful antioxidant, can effectively ameliorate the in vitro development of animal embryos, but few studies have been made on human embryos. Therefore, we investigated whether the application of MT in embryo cultures can improve embryonic development and clinical outcomes in patients with repeated cycles after failed in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) cycles.Methods: Human immature oocytes from controlled ovarian hyperstimulation cycles were collected for in vitro maturation culture and ICSI fertilization. The obtained embryos were cultured in vitro in medium containing 0, 10-11, 10-9, 10-7 or 10-5 M MT, and 10-9 M was determined to be the optimal concentration. Subsequently, 140 patients who experienced failed IVF/ICSI cycles received 140 cycles of embryo culture in vitro with medium containing 10-9 M MT. High-quality blastocysts were collected and cryopreserved for three months before vitrified-warmed embryo transfer. These culture cycles served as the experimental (10-9 M) group. The control (0 M) group comprised previous failed cycles. The patients were further divided into subgroups of 1, 2 or ≥3 failed cycles. The fertilization and embryo development status were compared.Results: The fertilization, cleavage, high-quality embryo, blastocyst, and high-quality blastocyst rates of the 10-9 M group were significantly higher than 0 M (87.7% vs. 83.6%, p p p Conclusion: The application of MT to embryo cultures in vitro improved embryonic development in patients with repeated cycles after failed IVF/ICSI cycles and obtained good clinical outcomes. The optimal concentration of MT was 10-9 M.Trial registration: Name of the registry: Effect of melatonin on clinical outcome of patients with repeated cycles after failed cycles of in vitro fertilization and intracytoplasmic sperm injection; registration number: ChiCTR2100045552; date of registration: April 19, 2021(Retrospectively registered); URL of trial registry record: www.medresman.org.cn.

Background: Preimplantation embryonic lethality is a rare cause of primary female infertility. Transducin-like enhancer of split 6 (TLE6) is a maternal effect gene which encodes a component of the subcortical maternal complex located in the oocytes and early embryos. It has been reported that biallelic variants in TLE6 can lead to preimplantation embryonic lethality. However, the incidence of TLE6 variants in patients with preimplantation embryonic lethality is not fully understood.Methods: Whole-exome sequencing and bioinformatics analyses were used to analyze a cohort of 28 patients with preimplantation embryonic lethality. We retrospectively analyzed the process and outcome of their attempts at in vitro fertilization and intracytoplasmic sperm injection. Results: In this study, four patients (14.29 %, 4/28) from three unrelated families in a cohort of 28 individuals with preimplantation embryonic lethality were identified as carrying biallelic TLE6 variants, including two homozygous variants and one compound heterozygous variant. These novel frameshift variants in TLE6 caused truncation of the TLE6 protein likely impairing its function. Immunofluorescence staining of oocytes for TLE6 indicated that it is localized in the oocyte cytoplasm, and that TLE6 was almost absent in the oocytes of the patients carrying biallelic TLE6 variants compared with normal control oocytes. A retrospective analysis showed that the four affected individuals underwent a total of nine in vitro fertilization and intracytoplasmic sperm injection attempts, in which a total of 119 metaphase II oocytes were retrieved, but none of them obtained high-quality blastocysts or obtained a successful pregnancy. However, one of these patients became pregnant on the first attempt using donated oocytes.Conclusions: Biallelic TLE6 variants cause preimplantation embryonic lethality that cannot be rescued by in vitro fertilization or intracytoplasmic sperm injection. Thus, oocyte donation may be the preferred treatment for patients harboring biallelic TLE6 variants.

Cryopreservation causes cryoinjury to oocytes and impairs their developmental competence. Melatonin (MLT) can improve the effect of cryopreservation in animal oocytes. However, no such studies on human oocytes have been reported. In this study, collected in vitro-matured human oocytes were randomly divided into the following groups: fresh group, MLT-treated cryopreservation (MC) group, and no-MLT-treated cryopreservation (NC) group. After vitrification and warming, viable oocytes from these three groups were assessed for their mitochondrial function, ultrastructure, permeability of oolemma, early apoptosis, developmental competence, and cryotolerance-related gene expression. First, fluorescence staining results revealed that oocytes from the 10-9 M subgroup showed the lowest intracellular reactive oxygen species and Ca2+ levels and highest mitochondrial membrane potential among the MC subgroups (10-11 , 10-9 , 10-7 , and 10-5 M). In subsequent experiments, oocytes from the 10-9 M-MC group were observed to maintain the normal ultrastructural features and the permeability of the oolemma. Compared with those of the oocytes in the NC group, the early apoptosis rate significantly decreased (P

Aim To compare embryonic developmental competence and clinical outcomes of oocytes matured in vivo (IVF oocytes) and those matured in vitro (IVM oocytes) from the same IVM/IVF cycles, and to analyze the clinical efficiency of a melatonin-supplemented in vitro maturation system combined with a modified IVM/IVF protocol. Main methods We randomly recruited 22 patients undergoing IVM/IVF treatment protocol in our medical centre. The fertilization, cleavage and blastocyst formation rates, as well as clinical pregnancy, implantation and live birth/ongoing pregnancy rates were analysed and compared between IVF and IVM oocytes. We evaluated mitochondrial function indicators by fluorescence staining and confocal microscopy, including mitochondrial membrane potential, reactive oxygen species and calcium (Ca2+) levels in 15 IVF and 15 IVM oocytes. Key findings There were no significant differences in fertilization or blastocyst formation rates between the IVF and IVM groups, whereas the cleavage rate was significantly higher in the IVF versus IVM group (100% vs 93.4 ± 10.9%, p = 0.03). There were no significant differences in the clinical pregnancy, implantation or live birth/ongoing pregnancy rates between the two groups. The cumulative clinical pregnancy and ongoing pregnancy/live birth rate per pick-up oocyte in the IVM/IVF treatment cycles were 68.2% (15/22) and 54.5% (12/22), respectively. The reactive oxygen species and Ca2+ levels were significantly increased, and mitochondrial membrane potential was significantly decreased, in IVM compared with IVF oocytes. Significance The modified IVM/IVF protocol can be effectively applied to the treatment of some indicated patients and achieve ideal clinical outcomes, even though the developmental potential of IVM oocytes may not be as high as IVF oocytes.

By application of preparative high-speed counter-current chromatography (HSCCC) to the crude quinolone alkaloids (1.1 g) from the fruit of Tetradium ruticarpum, 1-methyl-2((6Z,9Z)-pentadecadienyl)-4(1H)-quinolone (1, 8.4 mg), dihydroevocarpine (2, 27.0 mg), and 1-methyl-2-pentadecyl-4(1H)-quinolone (3, 18.8 mg) were isolated in one step with sufficient purity using the solvent system composed of hexane–ethyl acetate–methanol–water (Hex–EtOAc–MeOH–H2O, 5:2:5:3). Further purification of the subfraction was performed by amending the solvent composition and achieved another three quinolone alkaloids, i.e., 1-methyl-2-undecylquinolin-4(1H)-one (4, 13.7 mg), (Z)-1-methyl-2-(tridec-5-en-1-yl) quinolin-4(1H)-one (5, 14.0 mg) from subfraction FR3-A3-85 using Hex–EtOAc–MeOH–H2O (5:3.5:8.75:8.25), and 1-methyl-2-nonylquinolin-4(1H)-one (6, 15.1 mg) from subfraction FR3-A3-36 using Hex–EtOAc–MeOH–H2O (5:3.8:5:4.8). The relationship between the structure of the six alkaloids and their affinities for bovine serum album...

Melatonin (MT) regulates reproductive performance as a potent antioxidant; however, its beneficial effects on oocyte development remain largely unknown, especially in human oocytes. The collected 193 immature oocytes from the controlled ovarian hyperstimulation (COH) cycle underwent in vitro maturation (IVM) in IVM medium contained 10 μmol/L MT (n = 105, M group) and no MT (n = 88, NM group), followed by insemination and embryo culture. The results showed that the high-quality blastocyst formation rate in the M group (28.4%) was significantly higher than that in the NM group (2.0%) (P = .0001), and the aneuploidy rate was low (15.8%). In the subsequent clinical trials, three healthy infants were delivered. Next, single-cell RNA-seq data revealed 1026 differentially expressed genes (DEGs) between the two groups, KEGG enrichment analysis revealed that the majority of DEGs involved in oxidative phosphorylation pathway, which associated with ATP generation, was upregulated in the M group. Finally, confocal fluorescence staining results revealed that the mitochondrial membrane potential in the oocytes significantly increased and intracellular ROS and Ca2+ levels greatly decreased in the M group. Melatonin can promote the development of immature human oocytes retrieved from the COH cycle into healthy offspring by protecting mitochondrial function.

Does calcium ionophore treatment of oocytes improve fertilization rate, embryo development and outcomes in specific groups of infertile couples?This retrospective cohort study involved 796 couples undergoing oocyte activation with calcium ionophore (A23187) after intracytoplasmic sperm injection (ICSI) between 2016 and 2018. All metaphase II oocytes were exposed to 5 μmol/l ionophore for 15 min immediately after ICSI, cultured in vitro to the blastocyst stage, and transferred to the uteri of recipients on day 5 or cryopreserved for transfer in the next cycle. The previous cycles of the same patients formed the control group.Among 1261 ICSI cycles and 796 ICSI-artificial oocyte activation (ICSI-AOA) cycles, implantation, positive beta-HCG, clinical pregnancy and live birth rates were significantly (P0.05 to P0.001) improved for all groups, compared with previous cycles, except live birth rate in women with primary ovarian insufficiency (POI). Compared with previous cycles, rates of blastulation (all P0.001) and high-quality blastocysts (P0.05 to P0.001) were increased significantly for couples with male factor (oligoasthenoteratozoospermia [OAT]), unexplained infertility and couples with both factors in the ICSI-AOA cycles. High-quality blastocyst rate was increased in couples with polycystic ovary syndrome (PCOS) (P = 0.0453). Miscarriage rates were decreased significantly (P0.05 to P0.001) in couples with OAT, PCOS and unexplained infertility in the treatment cycles. No significant differences were found for fertilization rate, embryo development or live birth rate in patients with POI between both groups.Artificial oocyte activation was able to 'rescue' the poor reproductive outcomes in certain types of infertile couples with history of failure to achieve pregnancy.

Enhancement of the susceptibility of HL-60 cells to NK cell-mediated cytolysis induced by all-trans-retinoic acid (ATRA) plus valproate (VPA) was evaluated. In addition to the synergistic effect of ATRA plus VPA on HL-60 cells, the optimal concentration of 1 mM VPA plus 0.5 µM ATRA increased the cytotoxic sensitivity of HL-60 cells to NK cells. The expression of the activated receptors NKp30 and NKG2D on NK-92 cells was higher compared with the levels noted for the other receptors, and the expression of NKG2D ligands MICA/B on HL-60 cells was not significantly upregulated in the ATRA plus VPA goup compared with the control. Moreover, it was observed that the ligands of NKp30 on HL-60 cells presented the same variation trend. As to the co-stimulatory and adhesion molecules on NK-92 and their ligands on HL-60 cells post exposure to ATRA and VPA alone or their combination, there was no obvious change in the expression of CD112, CD48 and CD70 on the HL-60 cells. However, the expression of CD54 on HL-60 cells was significantly upregulated. In contrast, the expression of NKG2A ligands HLA-ABC on HL-60 cells was obviously downregulated. In addition, the expression of HLA-E on the HL-60 cells in the group treated with ATRA plus VPA was not significantly increased. In conclusion, the combination of VPA and ATRA not only induced the differentiation of HL-60 cells, but also induced enhancement of the sensitivity of HL-60 cells to NK cells by downregulating the expression of HLA-ABC and upregulating the expression of CD54, but not MICA/MICB. The results provide experimental and theoretical basis for the clinical combination of a low-dose of ATRA plus VPA for the treatment of leukemia.

A large number of recipients are in a compromised immune defense condition because of the routine application of immunosuppressive regimens after heart transplantation. Our previous work demonstrated that blockade of high-mobility group box 1 (HMGB1) prolongs the graft survival. Whether and how HMGB1 blockade impacts respiratory responses against pathogen-like challenge in organ transplant recipients when it improves cardiac graft are not elucidated. At the present study, after abdominal heterotopic heart transplantation, the recipient mice were treated with HMGB1 mAb, and then challenged with poly(I:C) or LPS intratracheally on day 7 post transplantation. We found that the level of bronchoalveolar lavage (BAL) HMGB1 was elevated after heart transplantation, and aggravated responses to respiratory tract poly(I:C)/LPS challenge were observed. HMGB1 neutralizing mAb treatment in poly(I:C)-challenged recipient mice alleviated pulmonary histopathological changes, neutrophil infiltration and inflammatory cytokine release, but unaffected the level of IFN-β, the distribution of CD11b(+)CD27(+)/CD11b(+)CD27(-) NK cell subsets, and CD8(+) T cell responses. In LPS-exposed recipient mice, HMGB1 mAb treatment ameliorated pulmonary inflammatory damage and enhanced the phagocytosis of phagocytic cells. Thus, this study may establish a basis for the application of HMGB1 blockade to improve the outcomes of heart transplant recipients because HMGB1 inhibition ameliorates pulmonary inflammation, but maintains defense-associated responses.

© The Author(s), 2016. This is the authors version of the work and is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Chemical Geology 423 (2016): 19-33, doi:10.1016/j.chemgeo.2016.01.003. Chromium (Cr) isotopes are an emerging proxy for redox processes at Earth’s surface. However, many geological reservoirs and isotope fractionation processes are still not well understood. The purpose of this contribution is to move forward our understanding of (1) Earth’s high temperature Cr isotope inventory and (2) Cr isotope fractionations during subduction-related metamorphism, black shale weathering and hydrothermal alteration. The examined basalts and their metamorphosed equivalents yielded δ53Cr values falling within a narrow range of -0.12±0.13‰ (2SD, n=30), consistent with the previously reported range for the bulk silicate Earth (BSE). Compilations of currently available data for fresh silicate rocks (43 samples), metamorphosed silicate rocks (50 samples), and mantle chromites (39 samples) give δ53Cr values of -0.13±0.13‰, -0.11±0.13‰, and -0.07±0.13‰, respectively. Although the number of high-temperature samples analyzed has tripled, the originally proposed BSE range appears robust. This suggests very limited Cr isotope fractionation under high temperature conditions. Additionally, in a highly altered metacarbonate transect that is representative of fluid-rich regional metamorphism, we did not find resolvable variations in δ53Cr, despite significant loss of Cr. This work suggests that primary Cr isotope signatures may be preserved even in instances of intense metamorphic alteration at relatively high fluid-rock ratios. Oxidative weathering of black shale at low pH creates isotopically heavy mobile Cr(VI). However, a significant proportion of the Cr(VI) is apparently immobilized near the weathering surface, leading to local enrichment of isotopically heavy Cr (δ53Cr values up to ~0.5‰). The observed large Cr isotope variation in the black shale weathering profile provides indirect evidence for active manganese oxide formation, which is primarily controlled by microbial activity. Lastly, we found widely variable δ53Cr (-0.2‰ to 0.6‰) values in highly serpentinized peridotites from ocean drilling program drill cores and outcropping ophiolite sequences. The isotopically heavy serpentinites are most easily explained through a multi-stage alteration processes: Cr loss from the host rock under oxidizing conditions, followed by Cr enrichment under sulfate reducing conditions. In contrast, Cr isotope variability is limited in mildly altered mafic oceanic crust. Funding for this research was provided by Agouron Institute to XLW, National Science Foundation (NSF) EAR-0105927 and EAR-1250269 to JJA, and NSF EAR-1324566 to ES. NJP and CTR acknowledge funding from the Alternative Earths NAI. 2017-01-12

Aims HMGB1 has been reported to play a crucial role in the physiological and pathophysiological responses during pregnancy. However, it is still unknown whether excessively expressed HMGB1 at the maternal-fetal interface related to Unexplained Recurrent Spontaneous Abortion (URSA). This study was designed to investigate the local capability of HMGB1 in the pathology of URSA, determined the distributions and characteristics of HMGB1, its receptors (RAGE/TLR2/TLR4) and important signaling molecule NF-κB p65 expression at the maternal-fetal interface，as well as compared the differences of HMGB1 expression between the URSA group, control group and aspirin treatment group. Material and methods H&E staining, Western blot analysis, immunofluorescence assay and immunohistochemical staining were applied to determine the effect of HMGB1 and its receptors at the maternal-fetal interface. ELISA was utilized to detect the concentration of HMGB1 in plasma. Key findings Our study demonstrated that the activation of the HMGB1-RAGE/TLR2/TLR4-NF-κB pathway at the maternal-fetal interface may have occurred in the URSA group. HMGB1 concentration in plasma was higher in the URSA group than the control group. Furthermore, the levels of HMGB1 of subjects with URSA could be reduced by administrating low doses of aspirin (ASPL). Significance This is the first report indicating the roles of HMGB1 at the maternal-fetal interface of URSA patients and broadening the horizons for clinical treatment of URSA. HMGB1-RAGE/TLR2/TLR4-NF-κB signaling pathway may be activated at the maternal-fetal interface in URSA and account for its pathogenesis. HMGB1 have the potential to be promising biomarkers in prevention and therapy of URSA.

Oxygenic photosynthesis fundamentally transformed all major biogeochemical cycles and increased the size and complexity of Earth's biosphere. However, there is still debate about when this metabolism evolved. As oxygenic photosynthesis is the only significant source of O-2 at Earth's surface, O-2 -sensitive trace element enrichments and isotopic signatures in Archean sedimentary rocks can potentially be used to determine the onset of oxygenic photosynthesis by tracking shifts in the oxidative capacity of Earth's surface environment. Here, we present an extensive new Archean U isotope record from iron formations, organic-rich shales, and paleosols. Variability in delta U-238 values gradually increased from Archean to Phanerozoic, consistent with current view of gradual oxidation of Earth's surface. In addition, statistical analysis on available delta U-238 data indicates a turning point of delta U-238 variability at roughly 3.0 billon years ago. We suggest that such a turning point in delta U-238 variability indicates the initiation of relatively large-scale oxidative weathering of U(IV)-bearing minerals, implying that oxygenic photosynthesis may have evolved before 3.0 billion years ago.

Interleukin-33 (IL-33) is usually expressed in the nucleus as a non-histone chromatin-associated protein. After passively released by necrotic cells, it functions as an IL-1 family member. IL-33 is highly expressed in the central nervous system (CNS), whether IL-33 is actively released in the CNS and involved in experimental autoimmune encephalomyelitis (EAE) remains unclear. In this study, we found that IL-33 and receptor ST2 were expressed in the spinal cord of naïve mice. Compared to naive situation, the intracellular IL-33 was dramatically decreased and extracellular IL-33 was markedly increased in the spinal cord in the pre-onset, onset and peak stage of EAE. In the chronic stage, the reverse happened. The decrease of intracellular IL-33 was related to the activation of astrocytes and the damage of neurons in situ during EAE. Astrocytes secreted IL-33 actively upon inflammatory stimulation in vitro. Furthermore, blockage of the CNS-derived IL-33 exacerbated EAE development. Our data demonstrated that IL-33 was released by activated astrocytes actively, and by damaged neurons during EAE. It plays a suppressive role in EAE development via an autocrine or paracrine manner. Our findings are helpful to understand the release feature and function of the CNS-derived IL-33 and supply a potential therapeutic target for multiple sclerosis.

The alkaloid rutaecarpine and its derivatives have been described as cytotoxic and hold potential as antitumor agents. Nevertheless, their synthesis is demanding and compounds display poor water solubility. Herein, we describe the synthesis of two sets of rutaecarpine derivatives with amine functions to improve solubility. Using a classic shake-flask experiment and a potentiometric titration platform, the water solubility of the compounds was determined. Solubility improved significantly with the amine functions connected over the indole-N atom. Reduction of metabolic activity and cell viability on HeLa cells was in the same range or better for these derivatives compared to the chemically unaltered parent compounds prepared in a new synthetic procedure established in our group.

Eurycoma longifolia is a tropical plant of diverse applications in folk medicine, which occurs in Southeast Asia. In this study, pre-purified fraction (0.86g) of the crude extracts from the roots of E. longifolia, was subjected to preparative high-speed counter-current chromatography (HSCCC) with a two-phase solvent system composed of hexane-ethyl acetate-methanol-water (HEMWat) at a volume ratio of 5:2:5:2 (v/v). Longifolione A (1, 19mg, purity 96.0%) and longifolione C (3, 317mg, purity 96.2%), together with longifolione B (2, purity 77.6%) were isolated in one run. The whole mobile and stationary phase was then blown out, concentrated in vacuo, and subjected to second HSCCC purification. Using HEMWat at a volume ratio of 6:1:6:1.2 (v/v), this fraction yielded two more new polyacetylenenes, longifolione D (4, 5mg purity 94.5%) and longifolione E (5, 33mg purity 96.3%). All of these five compounds are new natural products and isolated from E. longifolia for the first time. The established protocol for large-scale isolation of these polyacetylenes from E. longifolia was simple, efficient, and economical.

NK cells participate in the development of human multiple sclerosis (MS) and mouse experimental autoimmune encephalomyelitis (EAE), but the roles of different NK cell subsets in disease onset remain poorly understood. In this study, murine NK cells were divided into CD27high and CD27low/− subsets. The CD27high subset was decreased and the CD27low/− subset was increased in lymphoid organs during the pre-onset stage of EAE. Compared with the counterpart in naïve mice, the CD27high subset showed lower expression of Ly49D, Ly49H and NKG2D, and less production of IFN-γ, whereas the CD27low/− subset showed similar expression of the above mentioned surface receptors but higher cytotoxic activity in EAE mice. Compared with the CD27high subset, the CD27low/− subset exhibited increased promotion of DC maturation and no significant inhibition of T cells proliferation and Th17 cells differentiation in vitro. Additionally, adoptive transfer of the CD27low/− subset, but not the CD27high subset, exacerbated the severity of EAE. Collectively, our data suggest the CD27 NK cell subsets play different roles in controlling EAE onset, which provide a new understanding for the regulation of NK cell subsets in early autoimmune disease.

High mobility group box 1 (HMGB1), a nonhistone chromatin associated protein, plays different roles according to the expression pattern such as the amount, cell location and sub-cellular location. It has been recently demonstrated that the systemic HMGB1 is associated with autoimmune encephalomyelitis. However, the dynamic change of HMGB1 expression pattern in spinal cords that may be involved in the progression of disease is not fully understood. In this study, the amount, cell location and subcellular location of HMGB1 in adult mice spinal cords during various stages of experimental autoimmune encephalomyelitis (EAE) are investigated. HMGB1 is expressed in the nuclei of spinal cord resident cells such as some astrocytes, microglia and a few neurons in normal situation. During EAE progression, the total and extracellular HMGB1 in the spinal cord are increased, more HMGB1 positive astrocytes and microglia are observed, and the intra-neurons HMGB1 in the ventral horn and around the central canal localize majorly in the cytoplasm accompanied by the increasing extracellular HMGB1. Blockade of HMGB1 in central nervous system (CNS) locally attenuates the severity of EAE significantly. Our findings indicate that the HMGB1 expression pattern in the spinal cord is associated with the progression of EAE. HMGB1 may be a potential target for autoimmune encephalomyelitis (multiple sclerosis in human) therapy.

Blockade of extracellular high mobility group box 1 (HMGB1) can significantly prolong murine cardiac allograft survival. Here, we determined the role of HMGB1 in xenotransplantation. Sprague-Dawley rat hearts were transplanted heterotopically into BALB/c mice. Xenografts without any treatment developed predominant acute vascular rejection within 6 days. Both passively released HMGB1 from xenografts and actively secreted HMGB1 from infiltrated immune cells were significantly increased after xenotransplantation. HMGB1-neutralizing antibody treatment significantly prolonged xenograft survival and attenuated pathologic damage, immune cell infiltration, and HMGB1 expression and release in the xenografts. Compared to control IgG treatment evaluated at study endpoint, treatment with HMGB1-neutralizing antibody markedly suppressed xenoreactive B cell responses, as evidenced by the significant inhibition of anti-rat antibody production and deposition in xenografts at Day 6 posttransplant. Furthermore, treatment with anti-HMGB1 antibody suppressed B cell activation and reduced IFN-γ and IL-17A production after xenotransplantation. These results demonstrate for the first time that HMGB1 plays an important role in mediating acute xenograft rejection. Thus, we have shown that neutralization of extracellular HMGB1 can significantly inhibit xenoreactive B cell responses and delay xenograft rejection in a rat-to-mouse model of xenotransplantation, uncovering new insights in the role of HMGB1 in transplantation.

BACKGROUND High-temperatures in the cockpit environment can adversely influence pilot behavior and performance. OBJECTIVE To investigate the impact of high thermal environments on Chinese pilot performance in a simulated cockpit environment. METHODS Ten subjects volunteered to participate in the tests under 40°C and 45°C high-temperature simulations in an environmentally controlled chamber. Measures such as grip strength, perception, dexterity, somatic sense reaction, and analytical reasoning were taken. The results were compared to the Combined Index of Heat Stress (CIHS). RESULTS CIHS exceeded the heat stress safety limit after 45 min under 40°C, grip strength decreased by 12% and somatic perception became 2.89 times larger than the initial value. In the case of 45°C, CIHS exceeded the safety limit after only 20 min, while the grip strength decreased just by 3.2% and somatic perception increased to 4.36 times larger than the initial value. Reaction and finger dexterity were not statistically different from baseline measurements, but the error rate of analytical reasoning test rose remarkably. CONCLUSION Somatic perception was the most sensitive index to high-temperature, followed by grip strength. Results of this paper may help to improve environmental control design of new fighter cockpit and for pilot physiology and cockpit environment ergonomics research for Chinese pilots.

Chronic rejection that leads to diffuse narrowing and occlusion of graft vessels is the most important cause of morbidity and mortality following cardiac transplantation. The role and underlying mechanism of high-mobility group box 1 (HMGB1), as an established inflammatory mediator in acute rejection, remains poorly understood in chronic rejection. Here, we assessed the effects and mechanisms of HMGB1 on the chronic rejection using single MHC Class II-mismatched mouse cardiac transplantation model. It was found that HMGB1 was increased accompanying with the development of chronic rejection, while blockade of HMGB1 with specific neutralizing mAb substantially ameliorated chronic rejection-mediated vasculopathy and fibrosis of allograft, as well as markedly decreased T cell infiltration and production of IL-17A and interferon-gamma in allograft and recipient's spleen. Further, anti-HMGB1 antibody treatment significantly declined the number and frequency of mature dendritic cells (DCs) in allograft and recipient's spleen, especially CD11b(+) Ly6C(high) matured DCs that share the phenotypes with inflammatory-DCs. These findings indicate that HMGB1 contributes to chronic rejection, and HMGB1 blockade may be a novel mean to disrupt the proinflammatory loop after heart transplantation.