Desymmetrization-Addition Reaction of Cyclopropenes to Imines via Synergistic Photoredox and Cobalt Catalysis.

Herein, we designed a reaction for the desymmetrization-addition of cyclopropenes to imines by leveraging the synergy between photoredox and asymmetric cobalt catalysis. This protocol facilitated the synthesis of a series of chiral functionalized cyclopropanes with high yield, enantioselectivity, and diastereoselectivity (44 examples, up to 93% yield and >99% ee). A possible reaction mechanism involving cyclopropene desymmetrization by Co-H species and imine addition by Co-alkyl species was proposed. This study provides a novel route to important chiral cyclopropanes and extends the frontier of asymmetric metallaphotoredox catalysis.

Prenatal and postnatal exposure to heavy metals in PM2.5 and autism spectrum disorder.

Autism spectrum disorder (ASD) is a group of neurodevelopmental disorders, and its incidence is increasing over time. Although several environmental factors have been suspected to be risk factors for ASD, studies on the effects of airborne heavy metals on newly developed ASD are still limited. We conducted a large birth cohort study of 168,062 live term births in Taichung during 2004-2011 to assess the association of heavy metals in particulate matter with an aerodynamic diameter less than 2.5 µm (PM2.5) with ASD, and identify sensitive time windows during prenatal and postnatal periods. Heavy metals, including arsenic (As), cadmium (Cd), mercury (Hg), and lead (Pb) in PM2.5, were estimated using the Weather Research and Forecasting/Chem (WRF/Chem), inserted from the top 75 emission sources for the module. The association between childhood ASD and 4 metals were analyzed from pregnancy to 9 months after birth. The Cox proportional hazard model with a distributed lag nonlinear model (DLNM) was used to estimate the association between heavy metals in PM2.5 and ASD. We identified 666 incident ASD cases in 168,062 participants. A positive association between Hg and ASD was found at 9 months after birth (Hazard Ratio: 1.63; 95% CI: 1.13-2.36). According to the DLNM, there was an increased risk of exposure to Hg during 10-25 weeks after birth, and decreased risk of exposure to Hg during gestational weeks 4-6. Exposure to As and Hg on the risk of ASD were significantly stronger in low birth weight infants (<2500 g) than in those of birth weight ≥2500 g during postnatal period. Postnatal exposure to Hg in PM2.5 may associate with increased ASD incidence. Infants with low birth weight and exposure to As and Hg in PM2.5 are more likely to develop ASD.

The Correlation Between Aldosterone and Leukocyte-Related Inflammation: A Comparison Between Patients with Primary Aldosteronism and Essential Hypertension.

Background: Hypertension patients with primary aldosteronism (PA) have a higher risk of cardiovascular complications than blood pressure-matched essential hypertension (EH) patients. The cause may be closely related to inflammation. We explored the correlations between leukocyte-related inflammation parameters and plasma aldosterone concentration (PAC) in PA patients and clinical characteristics-matched EH patients. Methods: A total of 346 PA and 346 sex, age and 24-h blood pressure-matched EH patients at the 2nd Affiliated Hospital of Nanchang University from January 2020 to June 2021 were enrolled in this study. The differences and correlations of aldosterone and leukocyte parameters between the two groups were analyzed. Results: Compared with EH patients, the lymphocyte count was significantly lower (P = 0.004), the neutrophil-lymphocyte ratio (NLR) (P = 0.023) and the monocyte-lymphocyte ratio (MLR) (P = 0.037) were significantly higher in PA patients. Linear regression analysis and multivariate regression analysis identified that lymphocyte count, NLR and MLR were significantly and independently correlated with PAC in PA patients, and the correlations were stronger with increasing levels of aldosterone. However, in EH patients, only NLR maintained an independent correlation with PAC. Conclusion: Leukocyte-related inflammation parameters, including lymphocyte count, NLR, and MLR, were significantly and independently correlated with PAC in PA patients. The correlations were stronger with increasing levels of aldosterone. However, the above correlations were not always present in patients with EH matched for clinical characteristics.

The m6A reader MhYTP2 regulates the stability of its target mRNAs contributing to low nitrogen tolerance in apple (Malus domestica).

Studies have shown that the m6A reader primarily affects genes expression by participating in the regulation of mRNA localization, splicing, degradation, translation, and other metabolic processes. Previously, we discovered that the apple (Malus domestica) m6A reader MhYTP2 bound with and destabilized m6A-modified MdMLO19 mRNA. In addition, it enhanced the translation efficiency of m6A-modified mRNA of MdGDH1L, encoding a glutamate dehydrogenase, which confers resistance to powdery mildew. In this study, we report the function of MhYTP2 in the regulation of resistance to low nitrogen (N). The overexpression of MhYTP2 enhances the resistance of apple to low N. We show that MhYTP2 binds with and stabilizes the mRNAs of MdALN, which participates in the allantoin catabolic process and cellular response to N starvation in apple; MdPIDL, which participates in root hair elongation; MdTTG1, which is involved in the differentiation process of trichomes; and MdATG8A, which is a core participant in the regulation of autophagy. In addition, MhYTP2 accelerates the degradation of MdRHD3 mRNA, which regulates root development. RNA immunoprecipitation-seq and electrophoretic mobility shift assays show that the mRNAs of MdALN, MdATG8A, MdPIDL, MdTTG1, and MdRHD3 are the direct targets of MhYTP2. Overexpressing or knocking down the above genes in MhYTP2 overexpressing plants dismisses the function of MhYTP2 under low N, suggesting the role of MhYTP2 is dependent on those genes. Together, these results demonstrate that MhYTP2 enhances the resistance of apple to N deficiency by affecting the stability of the bound mRNAs.

The m6 A reader MhYTP2 negatively modulates apple Glomerella leaf spot resistance by binding to and degrading MdRGA2L mRNA.

Glomerella leaf spot (GLS), caused by the fungal pathogen Colletotrichum fructicola, significantly threatens apple production. Some resistances to plant disease are mediated by the accumulation of nucleotide-binding site and leucine-rich repeat (NBS-LRR) proteins that are encoded by a major class of plant disease resistance genes (R genes). However, the R genes that confer resistance to GLS in apple remain largely unclear. Malus hupehensis YT521-B homology domain-containing protein 2 (MhYTP2) was identified as an N6 -methyladenosine RNA methylation (m6 A) modified RNA reader in our previous study. However, whether MhYTP2 binds to mRNAs without m6 A RNA modifications remains unknown. In this study, we discovered that MhYTP2 exerts both m6 A-dependent and -independent functions by analysing previously obtained RNA immunoprecipitation sequencing results. The overexpression of MhYTP2 significantly reduced the resistance of apple to GLS and down-regulated the transcript levels of some R genes whose transcripts do not contain m6 A modifications. Further analysis indicated that MhYTP2 binds to and reduces the stability of MdRGA2L mRNA. MdRGA2L positively regulates resistance to GLS by activating salicylic acid signalling. Our findings revealed that MhYTP2 plays an essential role in the regulation of resistance to GLS and identified a promising R gene, MdRGA2L, for use in developing apple cultivars with GLS resistance.

Prenatal and postnatal exposure to PM2 .5 and the risk of tic disorders.

BACKGROUND: Tic disorders are common neurodevelopmental disorders during childhood. Whether prenatal and postnatal exposure to particulate matter with an aerodynamic diameter less than 2.5 µm (PM2.5 ) plays a role in the development of tic disorders remains unexplored. OBJECTIVES: To investigate the association of exposure between PM2.5 during the pregnancy and infancy periods and the risk of tic disorders. METHODS: This birth cohort study recruited singleton live births at term gestations in central Taiwan from the Taiwan Maternal and Child Health Database between 2004 and 2012 and followed up to the end of 2017. New cases of tic disorders were defined using the ICD-9-CM (307.2) and ICD-10-CM (F95), which include all tic spectrum disorders. We assigned daily PM2.5 concentrations derived from a satellite-based model to individuals based on maternal residential addresses at delivery. We fit Cox proportional hazard model and distributed lag non-linear model to estimate the associations between PM2.5 and tic disorders, with hazard ratio (HR) with 95% confidence interval (CI) as the effect measure. RESULTS: Of the 309,376 singleton live births at term gestations, we identified 5902 (1.9%) tic disorder cases. The HR of tic disorders was positively associated with a 10 µg/m3 increase in PM2.5 : during pregnancy HR 1.09, 95% CI 1.04, 1.15 and during infancy HR 1.12, 95% CI 1.06, 1.18. The vulnerable time window for infants with increased risk of tic disorders was 6-52 weeks after birth. We observed a nonlinear relationship between PM2.5 and the risk of tic disorders, with exposure to PM2.5 between 16 and 64 µg/m3 being associated with the risk of tic disorders. The association was restricted to Tourette's disorder group. Infant sex did not modify these associations. CONCLUSIONS: Infants delivered at term and exposed to PM2.5 are associated with an increased risk of tic disorders (6-52 weeks). Further studies are needed to confirm these associations.

Feature tuning improves MAXENT predictions of the potential distribution of Pedicularis longiflora Rudolph and its variant.

Pedicularis longiflora Rudolph and its variant (P. longiflora var. tubiformis (Klotzsch) Tsoong) are alpine plants and traditional Chinese medicines with important medicinal value, and future climate changes may have an adverse impact on their geographic distribution. The maximum entropy (MAXENT) model has the outstanding ability to predict the potential distribution region of species under climate change. Therefore, given the importance of the parameter settings of feature classes (FCs) and the regularization multiplier (RM) of the MAXENT model and the importance of add indicators to evaluate model performance, we used ENMeval to improve the MAXENT niche model and conducted an in-depth study on the potential distributions of these two alpine medicinal plants. We adjusted the parameters of FC and RM in the MAXENT model, evaluated the adjusted MAXENT model using six indicators, determined the most important ecogeographical factors (EGFs) that affect the potential distributions of these plants, and compared their current potential distributions between the adjusted model and the default model. The adjusted model performed better; thus, we used the improved MAXENT model to predict their future potential distributions. The model predicted that P. longiflora Rudolph and its variant (P. longiflora var. tubiformis (Klotzsch) Tsoong) would move northward and showed a decrease in extent under future climate scenarios. This result is important to predict their potential distribution regions under changing climate scenarios to develop effective long-term resource conservation and management plans for these species.

Comparison of Image Acquisition Techniques in Four-Dimensional Flow Cardiovascular MR on 3 Tesla in Volunteers and Tetralogy of Fallot Patients.

Four-dimensional phase-contrast (PC) velocity-encoded flow magnetic resonance imaging (4D flow MRI) is a potentially valuable tool for studying cardiovascular hemodynamics for disease monitoring and/or treatment planning. In this study we compared the performance of two 4D flow MRI pulse sequences - echo-planar imaging (EPI) and segmented gradient-echo (turbo-field-echo or TFE on vendor's platform) - on a clinical 3T system in 6 human subjects including 3 patients with Tetralogy of Fallot (TOF). For aortic flow rate, the coefficients of variation (COV) between 2D and 4D EPI were 7.0% and 7.7% for controls and patients respectively. The corresponding COV between 2D and 4D TFE were 19.0% and 18.3% for controls and patients respectively. The COV between 4D TFE and 4D EPI were larger than 18.7% in kinetic energy analysis. 4D EPI demonstrated acceptable accuracy of intra-cardiac flow quantification, which was also shown in the ex-vivo phantom measurements.

Development and evaluation of oral Cancer quality-of-life questionnaire (QOL-OC).

BACKGROUND: In this study scales and items for the Oral Cancer Quality-of-life Questionnaire (QOL-OC) were designed and the instrument was evaluated. METHODS: The QOL-OC was developed and modified using the international definition of quality of life (QOL) promulgated by the European Organization for Research and Treatment of Cancer (EORTC) and analysis of the precedent measuring instruments. The contents of each item were determined in the context of the specific characteristics of oral cancer. Two hundred thirteen oral cancer patients were asked to complete both the EORTC core quality of life questionnaire (EORTC QLC-C30) and the QOL-OC. Data collected was used to conduct factor analysis, test-retest reliability, internal consistency, and construct validity. RESULTS: Questionnaire compliance was relatively high. Fourteen of the 213 subjects accepted the same tests after 24 to 48 h demonstrating a high test-retest reliability for all five scales. Overall internal consistency surpasses 0.8. The outcome of the factor analysis coincides substantially with our theoretical conception. Each item shows a higher correlation coefficient within its own scale than the others which indicates high construct validity. CONCLUSIONS: QOL-OC demonstrates fairly good statistical reliability, validity, and feasibility. However, further tests and modification are needed to ensure its applicability to the quality-of-life assessment of Chinese oral cancer patients.

Stability of the V and Co atomic wires: a first-principles study.

We employ density-functional theory calculations plus pseudopotentials with the projector-augmented wave method to investigate the structural stability and electromagnetic characteristics of two infinite atomic wires made of vanadium (V) and cobalt (Co). We identify five stable V atomic wires and four stable Co atomic wires. The H structure of the V atomic wire shows semiconductor characteristics, and the other four structures show metallic properties. None of the V chains has magnetism. On the other hand, the four stable Co atomic wires have metal properties. The dimerized Co atomic chain is shown to be ferromagnetic with a maximum spin magnetic moment.

[Establishment of New Zealand rabbit models of aplastic anemia].

OBJECTIVE: To screen for the optimal dose of benzene and cyclophosphamide using an orthogonal design for establishment of New Zealand rabbit models of aplastic anemia. METHODS: Following an orthogonal experimental design, the effects of 3 levels of 4 factors, namely the dose of benzene (A), the dose of cyclophosphamide (B), the number of benzene injections (C), and the number of cyclophosphamide injections (D) were tested in the establishment of New Zealand rabbit models of aplastic anemia using a L9 (34) orthogonal table, and the optimal protocol for the model establishment was selected from the 9 experimental groups. Each rabbit received subcutaneous injection of benzene on the back every other day, followed by daily cyclophosphamide injection via the ear vein for prescribed times. The blood routine was examined every 6 days, and before modeling and at 36 days after modeling, a small sample of the femoral bone was collected for bone marrow histopathological examination. RESULTS: Comparison of the white blood cell, erythrocyte and platelet counts among the 9 groups showed successful modeling in Groups 4-9, and daily mean reduction rates of the cell counts in Groups 7, 8, and 9 differed significantly from those in the other groups (P<0.05). In Group 7, bone marrow sections showed low myelodysplasia, reduced hematopoietic tissue, reduced or even absence of megakaryocytes, and increased fat cells. Further observation found that the rabbits in Group 7 had sustained bone marrow suppression, consistent with the clinical characteristics of the disease. CONCLUSION: Stable models of aplastic anemia can be established efficiently in New Zealand rabbits by a combination of 8 subcutaneous injections of benzene at 1.5 mL/kg and 4 intravenous injections of cyclophosphamide at 10 mg/kg.

Fc-specific biotinylation of antibody using an engineered photoactivatable Z-Biotin and its biosensing application.

The development of a site-specific and covalent attachment methodology is crucial for antibody-biotin conjugates to preserve the antigen-binding ability of antibodies and yield homogeneous products. In this study, an engineered photoactivatable Z-domain variant [an UV-active amino acid benzoylphenylalanine (Bpa) was genetically incorporated into the Z-domain] carrying one biotin molecule (ZBpa-Biotin) was prepared by employing aminoacyl-tRNA synthetase/suppressor tRNA and Avitag/BirA techniques. The site-specific and covalent attachment of IgG-biotin conjugates, viz. photo-biotinylated IgG, was successfully achieved after UV exposure by combining the inherent Fc-binding capability of the Z-domain with the formation of covalent bond by the photo-crosslinker. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis assay showed that more than 90% of IgGs conjugated with ZBpa-Biotin molecules suffered 3 h UV irradiation. Further pepsin digestion analysis confirmed that the ZBpa-Biotin was conjugated to the Fc fragment of IgG without interference. We took the tumor biomarker carcinoembryoic antigen (CEA) as model to evaluate the detection efficiency of the site-specific photo-biotinylated IgG in biosensing application using surface plasmon resonance (SPR) technology. The photo-biotinylated IgG coated surface gave a limit of detection (LOD) of 2 ng mL-1, is 5-fold lower than that of the randomly NHS-biotinylated IgG (10 ng mL-1). Given that the (strept)avidin-biotin complex is extensively used in immunoassays, the proposed method for biotinylated IgG provides a powerful approach to further expand related applications.

Establishment of New Zealand rabbit models of aplastic anemia / 南方医科大学学报

<p><b>OBJECTIVE</b>To screen for the optimal dose of benzene and cyclophosphamide using an orthogonal design for establishment of New Zealand rabbit models of aplastic anemia.</p><p><b>METHODS</b>Following an orthogonal experimental design, the effects of 3 levels of 4 factors, namely the dose of benzene (A), the dose of cyclophosphamide (B), the number of benzene injections (C), and the number of cyclophosphamide injections (D) were tested in the establishment of New Zealand rabbit models of aplastic anemia using a L(3) orthogonal table, and the optimal protocol for the model establishment was selected from the 9 experimental groups. Each rabbit received subcutaneous injection of benzene on the back every other day, followed by daily cyclophosphamide injection via the ear vein for prescribed times. The blood routine was examined every 6 days, and before modeling and at 36 days after modeling, a small sample of the femoral bone was collected for bone marrow histopathological examination.</p><p><b>RESULTS</b>Comparison of the white blood cell, erythrocyte and platelet counts among the 9 groups showed successful modeling in Groups 4-9, and daily mean reduction rates of the cell counts in Groups 7, 8, and 9 differed significantly from those in the other groups (P<0.05). In Group 7, bone marrow sections showed low myelodysplasia, reduced hematopoietic tissue, reduced or even absence of megakaryocytes, and increased fat cells. Further observation found that the rabbits in Group 7 had sustained bone marrow suppression, consistent with the clinical characteristics of the disease.</p><p><b>CONCLUSION</b>Stable models of aplastic anemia can be established efficiently in New Zealand rabbits by a combination of 8 subcutaneous injections of benzene at 1.5 mL/kg and 4 intravenous injections of cyclophosphamide at 10 mg/kg.</p>

An efficient protocol to enhance the extracellular production of recombinant protein from Escherichia coli by the synergistic effects of sucrose, glycine, and Triton X-100.

Targeting recombinant proteins at highly extracellular production in the culture medium of Escherichia coli presents a significant advantage over cytoplasmic or periplasmic expression. In this work, a recombinant protein between ZZ protein and alkaline phosphatase (rZZ-AP) was constructed. Because rZZ-AP has the IgG-binding capacity and enzymatic activity, it can serve as an immunoreagent in immunoassays. However, only a very small portion of rZZ-AP is generally secreted into the aqueous medium under conventional cultivation procedure. Hence, we emphasized on the optimization of the culture procedures and attempted to dramatically enhance the yield of extracellular rZZ-AP from E. coli HB101 host cells by adding sucrose, glycine, and Triton X-100 in the culture medium. Results showed that the extracellular production of rZZ-AP in the culture medium containing 5% sucrose, 1% glycine, and 1% Triton X-100 was 18.6 mg/l, which was 18.6-fold higher than that without the three chemicals. And the ß-galactosidase activity test showed that the increased extracellular rZZ-AP was not due to cell lysis. Further analysis suggested a significant interaction effect among the three chemicals for the enhancement of extracellular production. Ultrastructural analysis indicated that the enhancement may be due to the influence of sucrose, glycine, and Triton X-100 on the periplasmic osmolality, permeability, or integrity of the cell wall, respectively. This proposed approach presents a simple strategy to enhance the extracellular secretion of recombinant proteins in the E. coli system at the process of cell cultivation.

Cardiovascular magnetic resonance reference ranges for the heart and aorta in Chinese at 3T.

BACKGROUND: Cardiovascular magnetic resonance (CMR) reference ranges have not been well established in Chinese. Here we determined normal cardiac and aortic reference ranges in healthy Singaporean Chinese and investigated how these data might affect clinical interpretation of CMR scans. METHODS: In 180 healthy Singaporean Chinese (20 to 69 years old; males, n = 91), comprehensive cardiac assessment was performed using the steady state free precision technique (3T Ingenia, Philips) and images were analysed by two independent observers (CMR42, Circle Cardiovascular Imaging). Measurements were internally validated using standardized approaches: left ventricular mass (LVM) was measured in diastole and systole (with and without papillary muscles) and stroke volumes were compared in both ventricles. All reference ranges were stratified by sex and age; and "indeterminate/borderline" regions were defined statistically at the limits of the normal reference ranges. Results were compared with clinical measurements reported in the same individuals. RESULTS: LVM was equivalent in both phases (mean difference 3.0 ± 2.5 g; P = 0.22) and stroke volumes were not significantly different in the left and right ventricles (P = 0.91). Compared to females, males had larger left and right ventricular volumes (P < 0.001 for all). Indexed LVM was significantly higher in males compared to females (50 ± 7 versus 38 ± 5 g/m2, respectively; P < 0.001). Overall, papillary muscles accounted for only ~2% of the total LVM. Indexed atrial sizes and aortic root dimensions were similar between males and females (P > 0.05 for all measures). In both sexes, age correlated negatively with left and right ventricular volumes; and positively with aortic sinus and sinotubular junction diameters (P < 0.0001 for all). There was excellent agreement in indexed stroke volumes in the left and right ventricles (0.1±5.7 mL/m2, 0.7±6.2 mL/m2, respectively), LVM (0.6±6.4 g/m2), atrial sizes and aortic root dimensions between values reported in clinical reports and our measured reference ranges. CONCLUSIONS: Comprehensive sex and age-corrected CMR reference ranges at 3T have been established in Singaporean Chinese. This is an important step for clinical practice and research studies of the heart and aorta in Asia.

Site-specific covalent attachment of an engineered Z-domain onto a solid matrix: an efficient platform for 3D IgG immobilization.

Immobilized antibodies with oriented and homogeneous patterns are crucial to solid-phase molecular recognition assay. Antibody binding protein-based immobilization can effectively present the desired antibodies. However, steadily installing the stromatoid protein with site-specific attachment manner onto a matrix surface remains to be elucidated. In this study, we present an optimal protocol to tightly attach an immunoglobulin G (IgG)-binding protein (Z-domain) through covalent incorporation of Cys-tag and maleimide group onto polystyrene surface to guarantee site-specific, oriented, and irreversible attachment, resulting in a highly efficient platform for three-dimensional IgG immobilization. The actual IgG-binding characteristic of immobilized Z-Cys was investigated by employing affinity chromatography and size exclusion chromatography. And the efficacy and potential of this platform was demonstrated by applying it to the analysis of interaction between rabbit anti-HRP IgG and its binding partner HRP. The proposed approach may be an attractive strategy to construct high performance antibody arrays and biosensors given that the antibody is compatible with the Z-domain.

[Effects of Emergent Water Project on vegetation coverage in the lower reaches of Tarim River].

Based on the TM in 1999, ETM in 2000, ASTER in 2004, and the background data of the lower reaches of Tarim River, the dynamic changes of vegetation coverage and land desertification before and after the implementation of Emergent Water Project were studied. The results showed that with the implementation of the Project, the areas with medium and higher vegetation coverage increased from 7,447.16 hm2 and 17,146.80 hm2 in 1999 to 9,129.16 hm2 and 26,236.61 hm2 in 2004, and those with lower and poor vegetation coverage decreased by 9,989.10 hm2 and 782.71 hm2, respectively. In 1990-2004, the total area of desertification decreased from 506,258.06 hm2 to 498,043.93 hm2, and the ecological environment in parts of the desertification area tended to be improved, indicating that the Emergent Water Project played an important role in improving the ecological environment in the lower reaches of Tarim River.