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Analyzing the pattern of altitudinal variation in the leaf traits and their networks of a particular tree species of similar age and its influencing factors could contribute to understanding the impacts of environmental factors on leaf traits and excluding the interference of genetic factors. We investigated the stomatal, structural, chemical, and vein traits of Daphniphyllum macropodum leaves in middle-aged forests, following the altitudinal gradient (1100, 1500, and 1900 m) on Mao'er Mountain. The objectives of this study were to reveal patterns in leaf trait and leaf trait networks variation, the life strategy of the tree species, and the major environmental factors affecting the altitudinal variations. The results showed that leaf area, specific leaf area, leaf thickness, leaf dry matter content, chlorophyll content, nitrogen content, phosphorus content, C:N, C:P, vein density, and vein diameter varied significantly across altitudes. Mean annual temperature and total radiation explained 42.1% and 16.2% of leaf-trait variation, respectively. They served as key environmental factors driving the altitudinal variation in leaf traits. Mean annual temperature exhibited the greatest influence on leaf area ( R 2 =0.73), and total radiation exerted the most prominent effect on leaf thickness ( R 2 =0.72). Both relationships were significantly positive. D. macropodum exhibited low leaf nitrogen and phosphorus at the low altitude of 1100 m, and the overall and local trait networks were loose, adopting a conservative resource strategy. At the medium altitude of 1500 m, leaf nutrient contents were relatively high. The overall network of leaf traits was tightly connected and local network was loose. By enhancing the dependency among leaf traits, and improving phosphorus utilization efficiency, D. macropodum could cope with competition in deciduous forests and adopt resource acquisition strategies. Further, at the highest altitude of 1900 m, D. macropodum had relatively large leaf thickness, chlorophyll content, and leaf dry matter content, but relatively small leaf area. The local network connections were tight while the overall network looseness, indicating a resource conserving strategy. The trade-off relationship between C:P and leaf phosphorus content was closely related to phosphorus use efficiency, and its variation was an important indicator for identifying life strategies of D. macropodum in different altitudes.

Only rarely have polyoxometalates been found to form core-shell nanoclusters. Here, we succeeded in isolating a series of rare giant and all-inorganic core-shell cobalt polyoxoniobates (Co-PONbs) with diverse shapes, nuclearities and original topologies, including 50-nuclearity {Co 12 Nb 38 O 132 }, 54-nuclearity {Co 20 Nb 34 O 128 }, 62-nuclearity {Co 26 Nb 36 O 140 } and 87-nuclearity {Co 33 Nb 54 O 188 }. They are the largest Co-PONbs and also the polyoxometalates containing the greatest number of Co ions and the largest cobalt clusters known thus far. These molecular Co-PONbs have intriguing and atomically precise core-shell architectures comprising unique cobalt oxide cores and niobate oxide shells. In particular, the encapsulated cobalt oxide cores with different nuclearities have identical compositions, structures and mixed-valence Co 3+ /Co 2+ states as the different sized Co-O moieties of the bulk cubic-spinel Co 3 O 4 , suggesting that they can serve as various molecular models of the cubic-spinel Co 3 O 4 . The successful construction of the series of the Co-PONbs reveals a feasible and versatile synthetic method for making rare core-shell heterometallic PONbs. Further, these new-type core-shell bimetal species are promising cluster molecular catalysts for visible-light-driven CO 2 reduction., (© 2023 Wiley-VCH GmbH.)

A rare cadmium-containing windmill-like heteropolyoxoniobate macrocycle has been successfully synthesized with stable 1-D cyclic cluster aggregates. The compound exhibited promising basic catalytic ability for Knoevenagel condensation with a high yield under mild reaction conditions and high cycling stability. The theoretical calculation showed that the promising basic catalytic ability is due to the dense and stronger basic sites of the surface terminal O atoms.

A new species of music frog, Nidiranaguibeiensis sp. nov. , is described from northern Guangxi, China. Based on two mtDNA fragments analyzed, phylogenetic trees reveal that N.guibeiensis sp. nov. is most closely related to N.leishanensis . However, the new species can be identified by conspicuous diagnostic morphological characteristics as well as bioacoustics. In contrast to the known Nidirana species, the advertisement calls of the new species can be divided into three types, calls with one, two, and three notes. In addition, the new species has nest construction behavior, which is inconsistent with N.leishanensis . Nidiranaguibeiensis sp. nov. occurs in paddy fields or still pools at 300-1300 m a.s.l., (Wei-Cai Chen, Jian-Ping Ye, Wan-Xiao Peng, Peng Li, Tong-Ping Su, Gui-Dong Yu, Zhi-Ying Cheng.)

Objective: To compare the effects of artesunate (Art) and fuzheng huayu decoction on mitochondrial autophagy in the treatment of schistosomiasis liver fibrosis. Methods: Eighty C57BL/6 female mice were randomly divided into healthy control group, infection group, Art treatment group and Fuzheng Huayu Decoction treatment group, with 20 mice in each group. Mice in the infection group and treatment group were infected with 16 Schistosoma japonicum cercariae. After 6 weeks, praziquantel (300 mg/kg) was used for 2 days to kill the worms. The Art treatment group was treated with intraperitoneal injection of 100 mg/kg/day, while the Fuzheng Huayu Decoction treatment group was fed 16g of fuzheng huayu decoction per 1kg per day. After 6 weeks, fresh liver tissues of the four groups were collected. Masson staining and Western blot were used to observe the succinate dehydrogenase subunit A (SDHA) and malate dehydrogenase (MDH2), citrate synthase (CS), ketoglutarate dehydrogenase (OGDH), and target of rapamycin 1 (mTORC1) pathway involved in mitochondrial tricarboxylic acid cycle in liver tissues. The relative expression levels of adenylate activated protein kinase (AMPK) and mitochondrial autophagy pathway kinase (PINK1) were detected. Liver tissue samples were extracted from each group to detect the mitochondrial oxygen consumption rate. Two-way ANOVA was used to compare the significance and difference between two sets of samples. Results: Masson staining showed that the infection group mice had significantly higher liver fibrosis area than the healthy control group, while the Art treatment group and Fuzheng Huayu Decoction treatment group mice had lower liver fibrosis area than the infection group. Western blot analysis showed that the infection group (0.82 ± 0.05) had significantly lower relative expression of SDHA protein than the healthy control group (1.00 ± 0.05) ( t = 11.23, P = 0.0035), while the Art treatment group (0.73 ± 0.05) had significantly higher relative expression of SDHA protein than the infection group ( t = 10.79, P = 0.0073). However, there was no significant change in Fuzheng Huayu Decoction treatment group (0.98±0.05) ( t = 1.925, P = 0.1266). The relative expression of p-AMPK protein was significantly higher in the infection group (1.15 ±0.05) than in the healthy control group (0.98 ± 0.07, t = 12.18, P = 0.0029), and the expression of p-AMPK in the Art treatment group (0.50 ± 0.05) was significantly lower than the infection group ( t = 11.78, P = 0.0032). The relative protein expression of AMPK was significantly lower in the infection group (0.80 ± 0.05) than in the healthy control group (1.00 ± 0.05, t = 10.53, P = 0.0046). The expression of AMPK was significantly lower in the Art treatment group (0.54 ± 0.05) than in the infection group ( T = 13.98, P = 0.0036). The relative expression of p-mTORC1 protein (0.93 ± 0.08) was not significantly different in the infection group than in the healthy control group ( t = 2.28, P = 0.065), while the Art treatment group (0.63 ± 0.05) had significantly lower relative expression of p-mTORC1 protein than the infection group ( t = 10.58, P = 0.029). The expression of p-mTORC1/ m-TORC1 was not significantly different in the infection group (0.98 ± 0.03) than in the healthy control group (0.97 ± 0.03, t = 0.98, P = 0.085), while the Art treatment group (0.63 ± 0.05) had significantly lower relative expression of p-mTORC1/ m-TORC1 than the infection group ( t = 14.58, P = 0. 009). The relative protein expression of PINK1 was significantly lower in the infection group (0.55 ± 0.05) than in the healthy control group (1.00 ± 0.03, t = 13.49, P = 0.0011), while the Art treatment group (1.21 ± 0.05, t = 9.98, P = 0.0046) and Fuzheng Huayu Decoction treatment group (1.31 ±0.35, t = 6.98, P = 0.027) had significantly higher relative protein expression of PINK1 than the infection group. Mitochondrial function tests showed that after adding substrate complex II, the oxygen consumption of the infection group was lower than the healthy control group, while the Art treatment group and the Fuzheng Huayu Decoction treatment group had higher oxygen consumption than the infection group. The oxygen consumption was significantly lower after adding the substrate complex III in the infection group than the healthy control group, while the Art treatment group and Fuzheng Huayu Decoction treatment group had higher oxygen consumption than the infection group. Conclusion: Art can alleviate schistosomiasis liver fibrosis by inhibiting AMPK/mTORC1 signaling pathway activity and enhancing mitochondrial oxygen consumption, autophagy and SDHA expression.

Mitochondrial response to inflammation is crucial in the metabolic adaptation to infection. This study aimed to explore the mitochondrial response under inflammatory and anti-inflammatory environments, with a focus on the tricarboxylic acid (TCA) cycle. Expression levels of key TCA cycle enzymes and the autophagy-related protein light chain 3b (LC3b) were determined in raw 264.7 cells treated with lipopolysaccharide (LPS) and metformin (Met). Additionally, reactive oxygen species (ROS) levels and mitochondrial membrane potential were assessed using flow cytometry. Moreover, 8-week-old C57BL/6J mice were intraperitoneally injected with LPS and Met to assess the mitochondrial response in vivo. Upon LPS stimulation, the expression of key TCA enzymes, including citrate synthase, α-ketoglutarate dehydrogenase, and isocitrate dehydrogenase 2, and the mitochondrial membrane potential decreased, whereas the levels of LC3b and ROS increased. However, treatment with Met inhibited the reduction of LPS-induced enzyme levels as well as the elevation of LC3b and ROS levels. In conclusion, the mitochondrial TCA cycle is affected by the inflammatory environment, and the LPS-induced effects can be reversed by Met treatment.

Keratin is widely used in the biomaterial application, but the keratin prepared by the physical or chemical approach has relatively low molecular weight and mechanical properties. Here we report the preparation of high molecular keratin (HMK) with molecular weight of 120 kDa via multi-enzyme cascade pathway and its application in wound healing. Briefly, we prepared the soluble keratin from wool by keratinase and improved the molecular weight of keratin by transglutaminase (TGase). The HMK was coelectrospun with poly(3-hydroxybutyric acid-co-3-hydroxyvaleric acid) (PHBV) and the prepared nanofibrous mats demonstrated improved mechanical properties. Ag nanoparticles (AgNPs) were synthesized on the nanofibers via in situ bioreduction, using the above-mentioned keratinase as the reducing agent. It is demonstrated that the PHBV/HMK/AgNPs nanofibrous mats possess favorable antibacterial properties and good biocompatibility. Moreover, in vivo wound healing assessment, the PHBV/HMK/AgNPs membrane displayed better wound healing ability than the control group. These results indicate that PHBV/HMK/AgNPs mats exhibit significant potential in tissue engineering., Competing Interests: Declaration of Competing Interest This manuscript has not been published and is not under consideration for publication elsewhere in whole or in part. We have no conflicts of interest to disclose. All authors have seen and approved the submission of the manuscript., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Keratin is widely recognized as a high-quality renewable protein resource for biomedical applications. Despite their extensive existence, keratin resources such as feathers, wool, and hair exhibit high stability and mechanical properties because of their high disulfide bond content. Consequently, keratin extraction is challenging and its application is greatly hindered. In this work, a biological extraction strategy is proposed for the preparation of bioactive keratin and the fabrication of self-assembled keratin hydrogels (KHs). Based on moderate and controlled hydrolysis by keratinase, keratin with a high molecular weight of approximately 45 and 28 kDa that retain its intrinsic bioactivities is obtained. The keratin products show excellent ability to promote cell growth and migration and are conferred with significant antioxidant ability because of their intrinsically high cysteine content. In addition, without the presence of any cross-linking agent, the extracted keratin can self-assemble into injectable hydrogels. The KHs exhibit a porous network structure and 3D culture ability, showing potential in promoting wound healing. This enzyme-driven keratin extraction strategy opens up a new approach for the preparation of keratin that can self-assemble into injectable hydrogels for biomedical engineering., (© 2020 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

ATP is an important energy source for cells. Traditionally, intracellular ATP levels are believed to be relatively stable and will not rise consistently in the physiological conditions. However, new studies suggest that ATP levels may rise in multiple tissues under the condition of energy surplus contributing to the metabolic disorders in obesity. However, the molecular mechanism of ATP elevation remains unknown in obesity except the increase in energy supply. Based on our experimental results and the findings reported in the literature, we discuss the cellular and molecular mechanisms by which ATP levels are regulated in cells by multiple factors, including superoxide ions, mitochondrial flash, antioxidants, anti-apoptotic molecule Bcl-xL, AMP-activated protein kinase (AMPK) and metformin. Contribution of these factors to the alteration of ATP set-point will be discussed together with their impact on insulin resistance in type 2 diabetes mellitus. With a focus on the energy surplus in obesity, we explore the mechanism of insulin resistance induced by ATP elevation and provide an answer to the contradiction between the new experimental results and the traditional viewpoint of intracellular ATP. We propose that elevation of intracellular ATP may lead to metabolic disorder in obesity through activation of a feedback mechanism that inhibits mitochondrial function.

Keratinases are promising alternatives over ordinary proteases in several industrial applications due to their unique properties compared with their counterparts in the protease categories. However, their large-scale industrial application is limited by the low expression and poor fermentation efficiency of keratinase. Here, we demonstrate that the expression level of keratinase can be improved by constructing a more efficient enzyme expression system hereby enables the highest production titer as regarding recombinant keratinase production to date. Specially, ten promoters were evaluated and the aprE promoter exhibits a significant promotion of keratinase (kerBv) titer from 165 U/mL to 2605 U/mL in Bacillus subtilis. The batch fermentation mode resulted in a maximum keratinase activity of 7176 U/mL at 36 h in a 5-L fermenter. Furthermore, the extracellular keratinase activity attained up to 16,860 U/mL via fed-batch fermentation within 30 h. The combination of keratinase with l-cysteine brings about 66.4 % degree of degradation of feather. Our work provides a new insight into the development of efficient keratinase fermentation processes with B. subtilis cell factory., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Fully-hydrogenated germanene, named germanane, represents a new nanostructured material for a variety of potential applications, such as electronics and optoelectronics. However, a critical requirement for developing practical and reliable electronic devices based on germanane consists of achieving a flexibly controllable charge carrier and doping level. Different to the conventional doping methods such as ion implantation and diffusion, by first-principles calculations we demonstrate that tetracyanobenzene (TCNB) molecular adsorption could introduce effective p-type doping in germanane due to the combination of germanane with electroactive acceptor molecule TCNB. The corresponding energy difference between the empty band minimum of the dopant and the valence band maximum for electron excitation is 0.173 eV. More importantly, this nondestructive p-type doping could be linearly tuned under an external E-field. Analysis of charge transfer by means of the equivalent capacitor model and the shift of energy levels in the superstructure of germanane/TCNB further reveals that the superposition of the external E-field and molecular adsorption-induced internal E-field plays a key role in the charge transfer between TCNB and germanane, especially in achieving a controllable p-type molecular doping level in germanane. Such convenient and flexible E-field-engineering of p-type molecular doping in germanane would be very helpful for potential applications of germanane-based electronic and optoelectronic devices in the future.

Three typical plant communities (evergreen broad-leaved forest at low-altitude 1100 m, evergreen and deciduous mixed broad-leaved forest at mid-altitude 1500 m, and evergreen conife-rous and broad-leaved mixed forest at high-altitude 1900 m) in Maoer Mountain, Guangxi, China were surveyed along an altitude gradient. We measured the tree layer plant architecture and environmental factors, to analyze the variation of plant architecture traits among the three communities and its influencing factors. The results showed that the tree layer canopy area, basal diameter at 45 cm height, diameter at breast height (DBH), and leaf convergence increased with increasing altitude, whereas tree height, branch height, and canopy thickness first increased and then decreased. Horizontal branches occurred more often in communities at lower altitude , less frequent at high altitude, and the least frequent in middle altitude communities. Correlations among tree layer plant architecture traits were stronger in the mid-altitude community than that in the other altitude communities. Results from the redundancy analysis showed that soil organic matter and total solar radiation were the main factors driving the variation of plant architecture traits in the tree layers, accounting for 39.6% and 23.9% of the total variation, respectively. Soil organic matter had a greater positive impact on canopy area and branch height, whereas total solar radiation was more influential on the DBH and 45 cm basal diameter. In conclusion, tree layer architecture of communities along the altitude gradient in Maoer Mountain was divergent, with soil organic matter and total solar radiation as the main driving forces.

To examine the change of runoff and its reason is an important scientific issue in forest hydrology. In this study, we performed the trend and inflection analysis on the time series of daily precipitation and runoff in the Pengchongjian small watershed from 1983 to 2014 by Mann-Kendall test method, a site with abundant precipitation . Using the empirical statistics method, we analyzed the effects of precipitation variation and vegetation restoration on the runoff and its contribution rates. Furthermore, we calculated the critical value of the hydrological effect of vegetation restoration on the annual precipitation. The results showed that the year 2003 was a consistent abrupt point for annual precipitation and runoff. Compared to the baseline period (1983-2003), annual precipitation and the depth of runoff in the changing period (2004-2014) decreased by 8.7% and 29.2%, with the averaged annual decrease of 12.7 and 22.1 mm, respectively. The averaged depth of runoff in spring, summer, autumn, winter, and in the whole year decreased by 100.2, 105.8, 25.2, 23.4, and 243.0 mm, respectively. The contribution rates of the depth of runoff to precipitation varia-tion were 58.9%, 71.6%, 65.5%, 35.0%, and 57.1%, respectively, while the contribution rates of vegetation restoration were 41.1%, 28.4%, 34.5%, 65.0%, and 42.9%, respectively. The hydrological effect of vegetation restoration was attributed to the annual precipitation, with a critical value of 1181 mm. Vegetation restoration increased annual depth of runoff when annual precipitation was less than the critical value, and increased that when annual precipitation was higher than the critical value. Therefore, the critical value might help to explain the difference in contribution rates of vegetation restoration to runoff in different watersheds and serve as one of the important reasons for the debate and divergence of forest restoration impacts on runoff.

Objective: To analyze the advantages of spatial measurement of anatomical parameters in a 3D model in surgical planning for laparoscopic partial nephrectomy (LPN)., Methods: From February, 2016 to October, 2017, 37 patients diagnosed with T1 renal mass underwent LPN based on 3D reconstruction after enhanced CT scanning using the Uromedix-3D system (group A), and another 38 patients received LPN with conventional CT planning (group B). The anatomical parameters were measured in the reconstructed 3D model and the demographic data, surgical outcome and postoperative data were compared between the two groups., Results: In group A, the average time for 3D model reconstruction was (29.3∓9.7) min; the length, width and depth of the renal defect in 3D model were 3.2∓1.1 cm, 2.6∓0.9 cm and 1.7∓0.7 cm, respectively; The distance of the tumor from the collecting system was 3.8∓2.2 mm; The mean R.E.N.A.L score of the patients was 7∓1.5, and 3 patients had accessory renal artery and 2 had early branching of the renal artery. LPNs were completed via the retroperitoneal approach in all the 75 patients without conversion to open or total nephrectomy. Group A and group B showed significant differences in warm ischemic time (26.7∓6.4 vs 31.9∓7.0 min), tumor-excision time (8.4∓2.6 vs 10.4∓2.8 min), renal defect suture time (18.3∓3.9 vs 21.5∓3.4 min), 24-h volume of retroperitoneal drainage (88.6∓40.2 vs 134.3∓58.3 mL) and 48-h volume of retroperitoneal drainage (127.9∓54.5 vs 198.1∓86.3 mL), but not in the demographic data, operation time, intraoperative blood loss or postoperative hospital stay., Conclusions: 3D reconstruction of the renal masses can be completed efficiently and accurately using this system. Compared with conventional CT-based measurement, 3D spatial measurement of the anatomical structures helps to increase the precision in the performance of LPN and reduce the warm ischemia time.

Background: Alpha-fetoprotein (AFP) has been shown to predict the prognosis of liver disease in several studies. This study aimed to evaluate the prognostic value of stratified AFP in patients with acute-on-chronic hepatitis B liver failure (ACHBLF)., Methods: A total of 192 patients were included and AFP were categorized into quartiles. The prognostic value was determined for overall survival (OS) and assessed by Kaplan-Meier analysis. Univariate and multivariate Cox proportional hazard regression analyses studied the association of all independent parameters with disease prognosis., Results: The optimal cut-off points of AFP were: (Q1) 252.3-4800.0 ng/ml, (Q2) 76.0-252.2 ng/ml, (Q3) 18.6-75.9 ng/ml, and (Q4) 0.7-18.5 ng/ml. Based on the Kaplan-Meier analysis of the OS, each AFP quartile revealed a progressively worse OS and apparent separation (log-rank P = 0.006). The second-highest quartiles of AFP (Q2) always demonstrated an extremely favorable short-term survival. Combining the lowest AFP quartiles with a serum sodium < 131mmol/L or an INR ≥ 3.3 showed a poor outcome (90-days survival of 25.0% and 11.9% respectively)., Conclusions: Stratified AFP could strengthen the predictive power for short-term survival of patients with ACHBLF. Combining AFP quartiles with low serum sodium and high INR may better predict poor outcome in ACHBLF patients.

Nanostarches were successfully prepared by high speed jet (HSJ) after pretreatment of micronization. The nanostarches were obtained at the conditions of micronization treatment for 60min, and then one cycle at 240MPa of HSJ (188.1nm). Moreover, after HSJ treated for three cycles, the particle size could reach the level of nanometer materials (66.94nm). The physicochemical properties of nanostarches had been characterized. Rapid Visco-Analysis (RVA) showed that the viscosity of nanostarches significantly decreased compared with native tapioca starch and slightly decreased with increasing processing cycles of HSJ. Steady shear analysis indicated that all samples displayed pseudoplastic, shear-thinning behavior, while the flow curves of nanostarches were little impact by the processing cycles of HSJ. X-ray diffraction analysis showed that the complete destruction of tapioca starch crystalline structure was obtained after HSJ treatment. Molecular characteristics determination suggested that the degradation of amylopectin chains occurred after the treatment of micronization and HSJ, which was proved by the decrease of weight-average molar mass. The results demonstrated that nanostarches were obtained due to the breakdown of starch molecules. This study will provide useful information of the nanostarches for its potential industrial application., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

A sensitive capsaicin sensor was constructed based on a poly(sodium 4-styrenesulfonate) functionalized graphite modified screen printed electrode (PSS-Grp/SPE) in this study. The PSS-Grp and poly(diallyldimethylammonium chloride) functionalized graphite (PDDA-Grp) were easily synthesized by interacting Grp with PSS and PDDA through sonication, and resulted in negative and with positive charges on the surface, respectively. The prepared PSS-Grp and PDDA-Grp were characterized by scanning electron microscopy (SEM), transmission electron microscopy (TEM) and ultraviolet and visible spectroscopy (UV-vis). The electrochemical performance of PSS-Grp in a 50 μM capsaicin solution presented a current density of 33 μA cm -2 , which was much higher than the PDDA-Grp of 1.5 μA cm -2 . Our study showed that capsaicin could interact better with strong negative charges on the PSS-Grp/SPE surface to give a higher electrochemical response. The direct electrochemical sensing of capsaicin was achieved at PSS-Grp/SPE using differential pulse stripping voltammetry (DPSV) under the optimized conditions.

BACKGROUND The endogenous protein annexin A1 (ANXA1) is an anti-inflammatory mediator in the brain that is thought to contribute to the progression of many neurological conditions. However, its exact role in temporal lobe epilepsy (TLE) remains unclear. We hypothesized that ANXA1 exerts negative actions on TLE by alleviating inflammatory damage in neurons. To identify the potential mechanism of TLE by assessing ANXA1 expression in TLE rats. MATERIAL AND METHODS TLE was induced in rats (n=70) via an intraperitoneal injection of lithium chloride (LiCl) and pilocarpine (PILO). The control group (n=10) received an injection of the equivalent amount of saline. ANXA1 expression was detected via immunohistochemistry and Western blotting. RESULTS Successful establishment of the TLE model in rats resulted in epileptic seizures. ANXA1 was immunohistochemically detected as brownish yellow particles in the dentate gyrus and the CA1 region of the door zone; this expression was predominantly localized to the cytoplasm of glia rather than neurons. ANXA1 expression was stronger in TLE rats compared with the control group. ANXA1 expression in TLE was also assessed via Western blotting, and compared between groups at various time points. ANXA1 expression was significantly increased in the acute (the first 24 h) and chronic (after 1 month) phases (P<0.001) but significantly decreased during the recovery phase (72 h, 1 week, and 2 weeks) (P<0.001). These findings suggest that ANXA1 expression is correlated with TLE activity. CONCLUSIONS Our data suggest that ANXA1 plays an important role in TLE by alleviating inflammatory damage and protecting neurons.

Five cis-clerodane diterpenoids, stephanialides A-E, along with seven known cis-clerodanes, scaparvins A-C, parvitexins B and C, 3-chloro-4-hydroxy-parvitexin A, and scapanialide B, were isolated from the Chinese liverwort Scapania stephanii. Their structures were established unequivocally on the basis of spectroscopic data. The absolute configuration of stephanialide A was determined by analysis of CD data using the octant rule. Phytotoxic activity evaluation showed that this type of diterpenoids can significantly inhibit root elongation of the seeds of Arabidopsis thaliana, Lepidium sativum and Brassica pekinensis., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Sixteen new clerodane diterpenoids, cephaloziellins A-P (1-16), and two known analogues (17 and 18) were isolated from an EtOH extract of the Chinese liverwort Cephaloziella kiaeri. The structures of the new compounds were elucidated from extensive spectroscopic data (IR, UV, HRESIMS, 1D NMR, and 2D NMR), and the structures of 5, 9, and 15 were confirmed by single-crystal X-ray diffraction analyses. The absolute configurations of all new compounds were established by comparing experimental and calculated electronic circular dichroism spectra.

Objective: To explore the role and mechanism of SIRT1 (Sirtuin1) in the differentiation of adipocyte., Methods: SIRT1(-/-) mice with C57BL/6J gene background were generated and litter-mate wild-type (WT) mice were used as controls. Body weight and fat content were detected and their epididymal fat pads were collected at 28 weeks old of age. The tip part of epididymal fat tissue was incubated in phosphate buffered saline (PBS) containing both BODIPY 558/568 (5 µmol/L in PBS) for adipocytes and isolectin Alexa Fluor 488 (40 µg/ml in PBS) for endothelial cells overnight. The stained cells were then visualized under confocal microscope.Reconstruction of 3D data sets was accomplished with image processing software Imaris.Immunohistochemistry was used to detect the protein level of endothelial cell marker CD31.Hematoxylin and eosin staining was performed for fixed epididymal fat tissue. Mouse embryonic fibroblast (MEF) cells were prepared from 13-14 days embryos of SIRT1(-/-) or WT mice and differentiated into adipocytes. Then oil-red O staining was performed., Results: Compared with wild-type controls, both body weight (WT 42.1 g ± 1.6 g vs SIRT1(-/-) 25.4 g ± 1.0 g, P < 0.05) and epididymal fat mass (WT 13.4 g ± 1.0 g vs SIRT1(-/-) 7.8 g ± 0.5 g, P < 0.05) were much smaller in the SIRT1(-/-) mice. HE staining of fat tissue exhibited a significant reduction in adipocyte size and extracellular matrix in SIRT1(-/-) mice.However, the adipogenesis ability of MEF cells was significantly enhanced in vitro in SIRT1(-/-) MEF cells.Further study found that the density of vascular network decreased by 50% in the tip portion of epididymal fat pads of SIRT1(-/-) mice (capillary density:WT 2.92% ± 0.03% vs SIRT1(-/-) 1.34% ± 0.02%, P < 0.05). CD31, a cellular marker of decreased angiogenesis, decreased significantly in epididymal fat pads of SIRT1(-/-) mice., Conclusion: SIRT1 knockout impairs adipocyte differentiation in SIRT1(-/-) mice with C57BL/6J gene background through reduced angiogenesis but not adipogenesis.

Chronic inflammation occurs in obese conditions in both humans and animals. It also contributes to the pathogenesis of type 2 diabetes (T2D) through insulin resistance, a status in which the body loses its ability to respond to insulin. Inflammation impairs insulin signaling through the functional inhibition of IRS-1 and PPARγ. Insulin sensitizers (such as rosiglitazone and pioglitazone) inhibit inflammation while improving insulin sensitivity. Therefore, anti-inflammatory agents have been suggested as a treatment strategy for insulin resistance. This strategy has been tested in laboratory studies and clinical trials for more than 10 years; however, no significant progress has been made in any of the model systems. This status has led us to re-evaluate the biological significance of chronic inflammation in obesity. Recent studies have consistently asserted that obesity-associated inflammation helps to maintain insulin sensitivity. Inflammation stimulates local adipose tissue remodeling and promotes systemic energy expenditure. We propose that these beneficial activities of inflammation provide an underlying mechanism for the failure of anti-inflammatory therapy in the treatment of insulin resistance. Current literature will be reviewed in this article to present evidence that supports this viewpoint.

Objective: To explore the effect of early insulin therapy on sterol regulatory element binding protein 1 (SREBP1) pathway and lipid accumulation in liver of type 2 diabetic rats (DM)., Methods: A high-fat diet plus a low-dose of streptozotocin (STZ) was administered to the Sprague-Dawley (SD) rats to create a type 2 diabetic animal model. Then the rats were divided into 3 groups: normal control (NC), DM (untreated diabetic rats) and INS (a 3-week treatment of NPH insulin initiated from day 3 of STZ injection). Insulin was delivered daily by a 3-week subcutaneous injection (6 - 8 U/day). Liver homogenate was prepared. The protein levels of ER stress marker immunoglobulin binding protein (Bip), oxygen-regulated protein 150 (ORP150), insulin-induced gene 1 (Insig1), SREBP1 and nuclear SREBP1 (nSREBP1) were assayed by Western blot. Adipose tissue mass was measured., Results: In the DM group, ER (endoplasmic reticulum) stress marker Bip and ORP150 were up-regulated (0.67 ± 0.02 vs 0.43 ± 0.01 for Bip; 1.11 ± 0.04 vs 1.83 ± 0.03 for ORP150, P < 0.05 for both) and Insig1 decreased (0.25 ± 0.02 vs 0.80 ± 0.07, P < 0.05). And the expressions of SREBP1 and nSREBP1 were elevated (1.03 ± 0.14 vs 0.41 ± 0.01 for SREBP1; 3.63 ± 0.77 vs 0.96 ± 0.20 for nSREBP1, P < 0.05 for both) in comparison with the normal control rats. In the INS group, all aforementioned changes became attenuated or reversed (0.41 ± 0.04 vs 0.67 ± 0.02 for Bip; 1.83 ± 0.03 vs 1.11 ± 0.04 for ORP150; 0.43 ± 0.02 vs 0.25 ± 0.02 for Insig1; 0.46 ± 0.01 vs 1.03 ± 0.14 for SREBP1; 1.65 ± 0.18 vs 3.63 ± 0.77 for nSREBP1, P < 0.05 for all). Furthermore, adipose tissue mass increased (22.4 g ± 3.6 g vs 12.0 g ± 2.6 g, P < 0.05)., Conclusion: The early insulin therapy induces a fat redistribution from liver to adipose tissue. The mechanism is probably through a reduction of ER stress and a down-regulated pathway of SREBP1 in liver of diabetic rats.

This study explored the feasibility of vertical insulin injection with an insulin pen injector in 40 Chinese diabetic outpatients with a normal body mass index. The patients, who received insulin in the hospital clinic, were assessed for abdominal subcutaneous fat thickness and distribution at four abdominal points using ultrasonography. Abdominal subcutaneous fat thickness and distribution were found to be heterogeneous and to differ significantly at these four points. Abdominal subcutaneous fat thickness was < 5 mm in nine of the 40 patients. In patients with abdominal subcutaneous fat thickness of < 5 mm, vertical insulin injection risks injecting into the muscle layer and is, therefore, not desirable. Vertical injection into pinched skin with a rotary syringe is safe and effective in such patients.

Background: NF-kappaB p65 was shown to inhibit transcription of phosphoenolpyruvate carboxykinase (PEPCK), a rate-limiting enzyme in gluconeogenesis in the liver. To understand the mechanism of action of NF-kappaB p65, we investigated the nuclear receptor corepressor in the regulation of PEPCK transcription., Methods: Rat H4IIE cells, human hepatoma HepG2 cells and human embryo kidney (HEK) 293 cells were used in this study. The transcriptional activity of a rat PEPCK gene promoter (-490/+100) was analyzed in HepG2 cells, a HepG2 super suppressor IkBalpha (ssIkBalpha) stable cell line, and HEK 293 cells. The effects of p65 and ssIkBalpha on a rat PEPCK gene promoter were observed using the PEPCK luciferase reporter system. The interaction of the cAMP-response- element-binding (CREB) protein, histone deacetylase 3 (HDAC3) and silencing mediator for retinoic and thyroid hormone receptors (SMRT) with the PEPCK gene promoter were investigated using the chromatin immunoprecipitation (ChIP) assay. p65 cotransfection and RNAi-mediated gene knockdown were used to determine the corepressor involved in the inhibition of PEPCK by NF-kappaB p65 and the transcriptional regulation of CREB by NF-kappaB p65., Results: NF-kappaB p65 inhibited PEPCK expression and the inhibition was blocked by ssIkBalpha. The inhibitory effect of p65 was completely blocked in a HepG2 stable cell line in which ssIkBalpha was expressed. HDAC3 or SMRT knockdown led to a significant up-regulation of PEPCK reporter activity in the presence of p65 cotransfection. In the ChIP assay the interaction of HDAC3 and SMRT with the PEPCK gene promoter was induced by p65 activation, but the CREB signal was reduced. Transcriptional activity of CREB was inhibited by NF-kappaB p65 cotransfection. The inhibitory effect of NF-kappaB p65 was blocked by HDAC3 RNAi or SMRT RNAi., Conclusions: The study showed that the inhibition of PEPCK by NF-kappaB p65 was dependent on HDAC3 and SMRT, which form a nuclear corepressor complex for transcriptional inhibition. The transcription factors NF-kappaB p65 and CREB share the same corepressor HDAC3-SMRT, and the corepressor exchange leads to inhibition of PEPCK gene transcription by NF-kappaB p65.

Fourteen solid coordination compounds of rare earth(Sm3+, Eu3+, Tb3+ and Dy3+ and Ln3+ (La3+, Gd3+ and Y3+)-doped Tb3+ with p-tert-butyl-calix[8] arene (LH8) have been synthesized and characterized. Elemental analysis, rare erth complexometry, 1H NMR spectra and TG-DTA have been studied. The results suggest that the complexes have the composition of [RE2 (LH2)(DMSO)5]DMSO, [RE2 (LH2) (DMF)5] 2DMF, [LnTb (LH2) (DMSO)5] DMSO and [LnTb (LH2) (DMF)5] 2DMF respectively. Fluorescence spectrum suggests that the Tb3+ complexes have characteristic luminescence, and its fluorescence intensity is enhanced after doping with La3+, Gd3+ and Y3+. The influence of the doping ions and two neutral ligands (DMSO and DMF) has also been discussed. In addition, the fluorescence lifetime of some complexes has been investigated.

In aqueous solution, the micellization and microenvironment characteristics of the micelle assemblies of three anionic surfactants, sodium 1-(n-alkyl)naphthalene-4-sulfonates (SANS), have been investigated by steady-state fluorescence and time-resolved fluorescence decay techniques using pyrene, Ru(bpy)3(2+), and 1,6-diphenyl-1,3,5-hexatriene as fluorescence probes. The critical micelle concentrations (cmc's), effective carbon atom numbers (neff's), hydrophilic-lipophilic balances (HLBs), mean micelle aggregation numbers, micropolarities, and microviscosities of these surfactant micelles have been determined. The logarithmic cmc of the alkylnaphthalene sulfonates decreases linearly with an increase in the neff. The logarithmic aggregation number of the alkylnaphthalene sulfonates increases linearly with an increase in the neff. However, in contrast to the alkylsufonates and the alkylbenzene sulfonates, the aggregation for these alkylnaphthalene sulfonate molecules is less sensitive to the increase in the neff. The micropolarity of these alkylnaphthalene sulfonate micelles is less sensitive to the increase in the alkyl chain length and is lower than that of sodium dodecyl sulfate (SDS). The microviscosity of these alkylnaphthalene sulfonate micelles increases with an increase in the alkyl chain length and is lower than those of nonionic surfactants and zwitterionic surfactants. These results suggest that naphthyl rings have a notable effect on the micellization of SANS.

Photooxygenations of 1,2-, 1,3-, and 2,3-di- and 1,2,3-trisubstituted indolizines 1a-1f under different reaction conditions in methanol and acetonitrile have been investigated to establish the general reaction pattern and mechanism in indolizine photooxygenation in view of the influence of the ring substituents and substitution pattern. Photooxygenations of 1-acyl-2-phenylindolizines 1a and 1b and 1,3-dibenzoyl-2-phenylindolizine (1d) are self-sensitized, while those of 1-(p-nitrobenzoyl)-2-phenylindolizine (1c) and 2-phenyl-3-(p-chlorobenzoyl)indolizine (1e) need to be sensitized by rose bengal (RB) or methylene blue (MB). These reactions proceed via a singlet oxygen mechanism yet follow different pathways in methanol and in acetonitrile, with peroxidic zwitterion D (in methanol) and dioxetane E across the indolizine C2-C3 bond (in acetonitrile) as the intervening intermediates. Methanol trapping of the peroxidic zwitterion results in C3-N bond cleavage and pyrrole ring opening to give the corresponding (E)- and (Z)-3-(2-pyridinyl)-3-benzoylpropenoic acid methyl esters (2 and 3) and 4-(2-pyridinyl)-3-phenyl-5-aryl-5-hydroxyfuran-2-one (4) as products in methanol, while O-O bond homolysis of the dioxetane furnishes 3-(2-pyridinyl)-3-benzoyl-2-phenyloxirane-2-carboxaldehyde (6) and 1-(6-methyl-2-pyridinyl)-2-phenylethanedione (5) as products in acetonitrile. 3-Benzoyl-1-indolizinecarboxylic acid methyl ester (1f) is unreactive toward singlet oxygen; however, it could be photooxygenated under electron transfer conditions with 9,10-dicyanoanthracene (DCA) as a sensitizer. This reaction takes place by the combination of the indolizine cation radical with the superoxide anion radical (or molecular oxygen) to give the pyridine ring oxidized methyl 3-benzoyl-5-methoxy-8-hydroxy-1-indolizinecarboxylate (9f), dimethyl 2-(2-pyridinyl)fumarate (8f), and dimethyl 2-(2-pyridinyl)maleate (7f) as products.

The present study was undertaken to test the hypothesis that the toxicity and carcinogenicity of vanadium might arise from elevation of reactive oxygen species leading to activation of the transcription factor activator protein-1 (AP-1). The AP-1 transactivation response has been implicated as causal in transformation responses to phorbol esters and growth factors. To investigate the possible activity of vanadium in the activation of AP-1, we treated mouse epidermal JB6 P+ cells stably transfected with an AP-1 luciferase reporter plasmid with various concentrations of vanadate. This resulted in concentration-dependent transactivation of AP-1. Superoxide dismutase (SOD) and catalase inhibited AP-1 activation induced by vanadate, indicating the involvement of superoxide anion radical (O2-*), hydroxyl radical (*OH) and/or H2O2 in the mechanism of vanadate-induced AP-1 activation. However, sodium formate, a specific *OH scavenger, did not alter vanadate-induced AP-1 activation, suggesting a minimal role for the *OH radical. NADPH enhanced AP-1 activation by increasing vanadate-mediated generation of O2-*. N-acetylcysteine, a thiol-containing antioxidant, decreased activation, further showing that vanadate-induced AP-1 activation involved redox reactions. Calphostin C, a specific inhibitor of protein kinase C (PKC), inhibited activation of AP-1, demonstrating that PKC is involved in the cell signal cascades leading to vanadate-induced AP-1 activation. Electron spin resonance (ESR) measurements show that JB6 P+ cells are able to reduce vanadate to generate vanadium(IV) in the presence of NADPH. Molecular oxygen was consumed during the vanadate reduction process to generate O2-* as measured by ESR spin trapping using 5,5-dimethyl-L-pyrroline N-oxide as the spin trapping agent. SOD inhibited the ESR spin adduct signal, further demonstrating the generation of O2-* in the cellular reduction of vanadate. These results provide support for a model in which vanadium, like other classes of tumor promoters, transactivates AP-1-dependent gene expression. In the case of vanadium, AP-1 transactivation is dependent on the generation of O2-* and H2O2, but not *OH.

Left ventricular end-diastolic compliance (LVEDC) and transmitral flow velocities were measured in 19 patients with coronary artery disease associated with hypertension and 10 normal subjects by LV catheterization and pulsed-Doppler echocardiography. LVEDC was much lower in the patient than in the normal subjects (P less than 0.01). The data showed that abnormal Doppler diastolic function such as elevated late diastolic peak flow velocity (PVA) and decreased LVEDC occurred in the patients at the same time. In addition, the negative correlation of PVA with LVEDC observed in normal controls but not in patients suggested that in patients with impaired diastolic filling, factors other than the decreased LVEDC itself must also participate in the development of diastolic dysfunction.

Left ventricular (LV) diastolic filling was examined by Doppler echocardiography in 239 patients with essential hypertension and 100 normal subjects. The Doppler study showed an inverse correlation between age and early diastolic peak velocity (PVE, r-0.201, P less than 0.05) [and a positive correlation between age and late diastolic peak velocity (PVA, r = 0.202, P less than 0.05) in normal subjects. Isovolumic relaxation time and late diastolic filling time were prolonged, PVA, A/E and Ai elevated, PVE, E/A and Ei/Ai as well as total filling time decreased in patients with hypertension as compared with the values found in the normal subjects (P less than 0.05 to 0.01). The results showed definite impairement of LV diastolic function in hypertensive patients. PVA correlated positively with systolic blood pressure (r = 0.68, P less than 0.01) and modestly with left atrial dimension in the hypertensives. No significant differences were found in FS and EF between the two study populations, indicating that LV filling abnormalities may occur early in patients with hypertension, even at a time when systolic performance has not yet been affected.