Dual-Locked Probe with Activatable Sonoafterglow Luminescence for Precise Imaging of MET-Induced Liver Injury.

Metformin (MET) is currently the first-line treatment for type 2 diabetes mellitus (T2DM). However, overdose and long-term use of MET may induce a serious liver injury. What's worse, diagnosis of MET-induced liver injury remains challenging in clinic. Although several probes have been reported for imaging MET-induced liver injury utilizing upregulated hepatic H2S as a biomarker, they are still at risk of nonspecific activation in complex physiological environments and rely on light excitation with limited imaging depth. Herein, we rationally designed and developed a dual-locked probe, DPA-H2S, for precise imaging of MET-induced liver injury by H2S-activated sonoafterglow luminescence. DPA-H2S is a small molecule consisting of a sonosensitizer protoporphyrin IX (PpIX) and an afterglow substrate that is dual-locked with a H2S-responsive 2,4-dinitrobenzene group and a 1O2-responsive electron-rich double bond. When employing DPA-H2S for imaging of MET-induced liver injury in vivo, since the PpIX moiety can produce 1O2 in situ at the liver site under focused ultrasound (US) irradiation, the two locks of DPA-H2S can be specifically activated by the highly upregulated H2S at the liver injury sites and the in situ generated 1O2, respectively. Thus, the sonoafterglow signal of DPA-H2S is significantly turned on, enabling precise imaging of the MET-induced liver injury. In vitro results showed that, through H2S-activated sonoafterglow luminescence, DPA-H2S was capable of imaging H2S with good sensitivity and high selectivity and realized deep tissue imaging (∼20 mm, signal-to-background ratio (SBR) = 3.4). Furthermore, we successfully applied DPA-H2S for precise in vivo imaging of MET-induced liver injury. We anticipate that our dual-locked probe, DPA-H2S, may serve as a promising tool in assisting the diagnosis of MET-induced liver injury in clinics and informing the clinical utilization of MET in the near future.

Quantifying the direct effects of long-term dynamic land use intensity on vegetation change and its interacted effects with economic development and climate change in jiangsu, China.

Vegetation change reflects sensitive responses of ecosystem environment to global climate change as well as land use. It is well known that land use type and its transformation affect vegetation change. However, how the changes in land use intensity (LUI) within different land use types impact vegetation and the interactions with other drivers remain poorly understood. We measured the LUI of Jiangsu Province, China, within the main land use types in 1995, 2000, 2005, 2010, 2015 and 2018 by combining remote sensing-based land use data with representative county scale economic and social indicators. Structural equation models (SEMs) were built to quantify the influences of long term LUI on vegetation change interacting with economic development, climate change and topographical conditions in transformed land, cropland, rural settlements and urbanized land, respectively. Seventy percent of significant vegetation change existed in non-transformed land use types. Although the area with a vegetation greening trend is larger than that with a vegetation browning trend, the vegetation browning areas is prominent in urbanized lands and some croplands in south basins. The constructed SEMs suggested the dominant negative effect of fast economic development regardless of land use types, while LUI played important and different direct and indirect effects on affecting vegetation change significantly interacting with economic development and climate change in different land use types. The LUI increasing led a vegetation greening in cropland, and stronger than climate warming with both positive direct and indirect effects for influencing climate change. The LUI change took negative effects on vegetation change in rural and urban areas, while a positive indirect effect of LUI increasing in urbanized land signaled the positive results of human managements. We then provided some land use-specific suggestions on basin scale for land management in Jiangsu. Our results highlight the necessity of long-term LUI quantification and promote the understanding of its effects on vegetation change interacted with other drivers within different land use types. This can be very helpful for sustainable land use and managements in regions with fast economic development.

[Bladder wall neourethra technique improves early urinary continence after laparoscopic radical prostatectomy].

OBJECTIVE: To study the effect of the bladder wall neourethra (BWN) technique on early urinary continence after laparoscopic radical prostatectomy (LRP). METHODS: We prospectively selected 40 cases of LRP performed in our hospital from August 2020 to August 2021 and randomly divided them into a BWN group (n = 20) and a control group (n = 20). We recorded the urinary continence rate of the two groups of patients at 7, 30, 90 and 180 days, and measured the maximum urethral pressure (MUP), functional urethral length (FUL) and functional urethral area (UFA) and observed the shape of the neourethra closure by MRI at 1 month after catheter removal. RESULTS: The urinary continence rates were significantly higher in the BWN than in the control group at 7 days (90.0% vs 25.0%, P < 0.001), 30 days (95.0% vs 35.0%, P < 0.001), 90 days (100% vs 60.0%, P < 0.05) and 180 days (100% vs 90.0%, P > 0.05) after catheter removal. No statistically significant difference was observed in MUP between the two groups (P > 0.05). FUL and FUA were remarkably higher in the BWN than in the control group (P < 0.01). MRI showed tight closure of the neourethra in the BWN group in the urine storage period. CONCLUSION: The BWN technique can significantly prolong FUL and improve early urinary continence after LRP.

[Chemical constituents from the leaves of Cinnamomum camphora var. linaloolifera and their anti-inflammatory activities].

Thirteen compounds were isolated and purified from the leaves of Cinnamomum camphora by the macroporous resin,silica gel,and Sephadex LH-20 column chromatographies. Those compounds were further identified by IR,UV,MS,and NMR techniques:( 2 S)-1-( 3″,4″-methylenedioxy phenyl)-3-( 2',6'-dimethoxy-4'-hydroxyphenyl)-propan-2-ol( 1),( 2 R,3 R)-5,7-dimethoxy-3',4'-methylenedioxy flavanol( 2),9-hydroxysesamin( 3),sesamin( 4),piperitol( 5),kobusin( 6),(-)-aptosimon( 7),acuminatolide( 8),1ß,11-dihydroxy-5-eudesmene( 9),lasiodiplodin( 10),vanillin( 11),p-hydroxybenzaldehyde( 12),and p-hydroxybenzoic acid ethyl ester( 13). Compound 1 was a novel compound,and compounds 2,6,7,9 and 10 were isolated from Cinnamomum plants for the first time. Compounds 4,7 and 10 were found to possess good inhibitory effect on IL-6 production in LPS-induced BV2 cells at a concentration of 20 µmol·L-1 in the in vitro bioassay,with inhibition rates of 51. 26% ± 4. 13%,67. 82% ± 3. 77% and85. 81%±1. 19%,respectively.

Coexpression network analysis of platelet genes in sickle cell disease.

Platelets play important roles in vascular health. Activation of platelet may contribute to coagulation and inflammation. Evidence suggests circulating platelets are chronically activated in sickle cell disease (SCD) patients with steady state and further activated in vaso-occlusive crisis. However, the molecular basis of sickle platelet dysfunction remains obscure. Here, we used weighted gene coexpression network analysis combined with differentially expressed genes (DEGs) analysis to further investigate this basis. We found 57 DEGs were closely related to platelet dysfunction in SCD. Enrichment analysis showed that these 57 genes were mostly related to protein synthesis, adenosine triphosphate (ATP) synthase activity and inflammation, suggesting a hyperactivation status of platelets in SCD. We identified six hub genes from the 57 DEGs according to their Gene Significance value ranking, including CRYM, CCT6P1, SUCNR1, PRKAB2, GSTM3 and FCGR2C. Altogether, our results offered some new insight into platelet activation and identified novel potential targets for antiplatelet therapy in SCD.

Association of vitamin D receptor gene polymorphisms and serum 25-hydroxyvitamin D levels with Crohn's disease in Chinese patients.

BACKGROUND AND AIM: The vitamin D receptor (VDR) regulates immune responses and inflammation through binding with 1,25-dihydroxyvitamin D, the active form of vitamin D. The serum 25-hydroxyvitamin D (25(OH)D) level clinically reflects vitamin D status in the human body. We investigated the association of VDR polymorphisms and 25(OH)D levels in Chinese patients with Crohn's disease (CD). METHODS: Vitamin D receptor polymorphisms (FokI, BsmI, ApaI, and TaqI) were genotyped by SNaPshot. Serum 25(OH)D levels were measured by electro-chemiluminescence immunoassay. RESULTS: A total of 297 patients with CD and 446 controls were recruited. Compared with controls, mutant alleles and genotypes of BsmI and TaqI were less prevalent in patients with CD (all P < 0.05/4 = 0.0125). The AAC haplotype formed by BsmI, ApaI, and TaqI was also less prevalent in patients with CD (P = 0.004). Furthermore, 124 patients and 188 controls were randomly selected for measurements of 25(OH)D levels. Average 25(OH)D level was lower in patients with CD than in controls (15.46 ± 8.11 vs 21.64 ± 9.45 ng/mL, P < 0.001) and negatively linked to CD activity index (ß = -0.829, P < 0.001), platelet count (ß = -0.253, P < 0.001) and neutrophil percentage (ß = -0.136, P = 0.005) in patients with CD. The ApaI mutant genotype and vitamin D deficiency (<20 ng/mL) were independently associated with CD (P = 0.009, P < 0.001, respectively). In patients with CD, vitamin D deficiency interacted with FokI, ApaI, and TaqI mutant genotypes (P = 0.027, P = 0.024, and P = 0.040, respectively). CONCLUSIONS: Vitamin D receptor (BsmI, ApaI, and TaqI) mutations and lower 25(OH)D levels are associated with CD in Chinese patients. Moreover, VDR (FokI, ApaI, and TaqI) mutations and vitamin D deficiency may have a combined impact on CD.

Association of vitamin D receptor gene polymorphisms with the susceptibility to ulcerative colitis in patients from Southeast China.

The association studies from different ethnic groups showed that vitamin D receptor (VDR) gene polymorphisms might be connected with the susceptibility to ulcerative colitis (UC); however, the conclusions were less consistent. Our study aimed to analyze the associations of UC with common mutations of VDR in Chinese patients. A total of 382 UC patients and 489 healthy controls were recruited. The genotypes of VDR FokI (rs2228570), BsmI (rs1544410), ApaI (rs7975232) and TaqI (rs731236) were examined by SNaPshot assays. Haplotype analysis was performed in all study subjects. After Bonferroni correction, the mutant alleles and genotypes of VDR FokI, BsmI, ApaI and TaqI did not statistically differ between UC patients and the controls (all p > 0.0125). However, the mutant allele C and genotype TC + CC of FokI gene were significantly increased in patients with mild and moderate UC compared to those with severe UC (C allele: 54.1% versus 39.3%, OR = 1.83, 95% CI: 1.21-2.75, p = 0.004; TC + CC genotype: 81.6% versus 57.1%, OR = 3.32, 95% CI: 1.83-6.06, p < 0.001, respectively). Haplotype analysis showed that the VDR BsmI, ApaI and TaqI polymorphic loci were in a strong linkage disequilibrium. Furthermore, the frequency of AAC haplotype was statistically lower in UC patients than that in the controls (3.8 versus 5.9%, OR = 0.63, 95% CI: 0.39-1.01, p = 0.039). In conclusion, the mutation of FokI gene influenced severity of the disease in UC patients. Moreover, the AAC haplotype formed by the VDR BsmI, ApaI and TaqI gene might engender a reduced risk of UC attack.

Shadow Detection in Remote Sensing Images Based on Spectral Radiance Separability Enhancement.

Shadow detection is a basic task of remote sensing image analysis, but it is often seriously disturbed by vegetation, water bodies, and black objects. It is observed that vegetation and dark objects often show a dark look in visible bands but brighter in the near-infrared (NIR), and is also noticed that the reflection of inland water bodies in the green band is stronger than that in the blue band. Taking advantage of these physical properties and combining them with the bluish and dark appearance of shadows, we propose a simple but effective shadow detection method for multispectral remote sensing images. These physical properties are used to create transformation models that suppress features such as vegetation, water bodies, etc., but at the same time enhance shadows. Then, we transform the shadow representation into a color space to generate candidate shadows using dominant color components. To separate shadows from the others, we propose two indexes, the normalized Color Difference Composite Index (CDCI) and Color Purity Index (CPI), and fuse them to achieve shadows and their confidence. The experimental results indicate that the proposed method can effectively detect the shadows in multispectral images and outperforms the state-of-the-art approaches.

Insulin induces C2C12 cell proliferation and apoptosis through regulation of cyclin D1 and BAD expression.

Insulin is a secreted peptide hormone identified in human pancreas to promote glucose utilization. Insulin has been observed to induce cell proliferation and myogenesis in C2C12 cells. The precise mechanisms underlying the proliferation of C2C12 cells induced by insulin remain unclear. In this study, we observed for the first time that 10 nM insulin treatment promotes C2C12 cell proliferation. Additionally, 50 and 100 nM insulin treatment induces C2C12 cell apoptosis. By utilizing real-time PCR and Western blotting analysis, we found that the mRNA levels of cyclinD1 and BAD are induced upon 10 and 50 nM/100 nM insulin treatment, respectively. The similar results were observed in C2C12 cells expressing GATA-6 or PPARα. Our results identify for the first time the downstream targets of insulin, cyclin D1, and BAD, elucidate a new molecular mechanism of insulin in promoting cell proliferation and apoptosis.

Efficacy of Baishao Luoshi decoction on synaptic plasticity in rats with post stroke spasticity.

OBJECTIVE: To evaluate the efficacy of Baishao Luoshi decoction (, BD) on synaptic plasticity in rats with post stroke spasticity (PSS), and to study the mechanism behind the action. METHODS: The PSS model of rat was established by middle cerebral artery occlusion (MCAO). The neurological deficit symptoms were evaluated by modified neurological deficit score (mNSS). Muscle tension were evaluated by Modified Ashworth score (MAS). Transmission electron microscopy (TEM) was used to observe the synaptic ultrastructure. The expression of synaptic plasticity-related protein brain derived neurotrophic factor (BDNF), growth associated protein-43 (GAP43), synaptophysin (p38) and microtubule-associated protein 2 (MAP2) in the brain tissue around the infarct were detected by Western blotting. RESULTS: We found that mNSS were significantly improved and limb spasticity was ameliorated treated by BD. The thickness of postsynaptic density and the synaptic curvature increased significantly. The expression of synaptic plasticity-related protein BDNF, GAP43, p38, MAP2 in the brain tissue around the infarct were raised remarkably after treated by BD. CONCLUSIONS: Alleviating PSS by BD may be related to rescuing the synaptic plasticity, which provides a probable new therapeutic method for PSS.

Hyaluronic acid increases MMP-2 and MMP-9 expressions in cultured trabecular meshwork cells from patients with primary open-angle glaucoma.

PURPOSE: A depletion of hyaluronic acid (HA) in patients' eyes may be associated with primary open-angle glaucoma (POAG), but the exact mechanism remains unclear. We investigated the effect of HA on the expression of matrix metalloproteinases (MMP-2 and MMP-9) in cultured trabecular meshwork cells. METHODS: Trabecular meshwork cells were cultured from trabecular tissues obtained from the POAG patients aged 23 to 45. The mRNA expression of MMP-2 and MMP-9 was determined by reverse transcription-polymerase chain reaction, and the protein expression of MMP-2 and MMP-9 by gelatin zymography analysis and qualified by the gel electrophoresis image analysis in different HA concentrations. RESULTS: The expression of MMP-2 and MMP-9 by the two methods significantly increased with HA concentration in a dose-response manner. Mean values of the MMP-2 expression by the gelatin zymography analysis were 176, 264, 353, and 448 mg/ml, and mean values of the MMP-9 expression were 547, 659, 895, and 1,147 mg/ml, for HA concentration level of 0, 1, 3, and 6 mg/ml, respectively. CONCLUSIONS: In POAG trabecular meshwork cells, the level of HA concentration increases the activities of MMP-2 and MMP-9. The lack of HA in aqueous humor can result in a reduction in activities of MMPs and therefore may be involved in the pathogenesis of POAG.

Towards constructing multi-bit binary adder based on Belousov-Zhabotinsky reaction.

It has been proposed that the spatial excitable media can perform a wide range of computational operations, from image processing, to path planning, to logical and arithmetic computations. The realizations in the field of chemical logical and arithmetic computations are mainly concerned with single simple logical functions in experiments. In this study, based on Belousov-Zhabotinsky reaction, we performed simulations toward the realization of a more complex operation, the binary adder. Combining with some of the existing functional structures that have been verified experimentally, we designed a planar geometrical binary adder chemical device. Through numerical simulations, we first demonstrated that the device can implement the function of a single-bit full binary adder. Then we show that the binary adder units can be further extended in plane, and coupled together to realize a two-bit, or even multi-bit binary adder. The realization of chemical adders can guide the constructions of other sophisticated arithmetic functions, ultimately leading to the implementation of chemical computer and other intelligent systems.

[On-orbit response variation analysis of FY-3 MERSI reflective solar bands based on Dunhuang site calibration].

MERSI is the keystone payload of FengYun-3 and there have been two sensors operating on-orbit since 2008. The on-orbit response changes obviously at reflective solar bands (RSBs) and must be effectively monitored and corrected. However MERSI can not realize the RSBs onboard absolute radiometric calibration. This paper presents a new vicarious calibration (VC) method for RSBs based on in-situ BRDF model, and vector radiometric transfer model 6SV with gaseous absorption correction using MOTRAN. The results of synchronous VC experiments in 4 years show that the calibration uncertainties are within 5% except for band at the center of water vapor absorption, and 3% for most bands. Aqua MODIS was taken as the radiometric reference to evaluate the accuracy of this VC method. By comparison of the simulated radiation at top of atmosphere (TOA) with MODIS measurement, it was revealed that the average relative differences are within 3% for window bands with wavelengths less than 1 microm, and 5% for bands with wavelengths larger than 1 microm (except for band 7 at 2.1 microm). Besides, the synchronous nadir observation cross analysis shows the excellent agreement between re-calibrated MERSI TOA apparent reflectance and MODIS measurements. Based on the multi-year site calibration results, it was found that the calibration coefficients could be fitted with two-order polynomials, thus the daily calibration updates could be realized and the response variation between two calibration experiments could be corrected timely; there are large response changes at bands with wavelengths less than 0.6 microm, the degradation rate of the first year at band 8 (0.41 microm) is about 14%; the on-orbit response degradation is maximum at the beginning, the degradation rates slow down after one year in operation, and after two years the responses even increase at some band with wavelengths larger than 0.6 microm.

[Study on the optical properties of ultra-thin metal films in the THz band].

Thin metal films are good candidates of terahertz detectors, reflectors, waveguides and terahertz quantum-cascade lasers (THz-QCLs). The optical parameter is the basis not only for designing the THz components but also for developing novel optoelectronic materials. In the present paper, the complex refractive indices of the ultra-thin metal (Cr, Ni and Ti) films in the THz band were obtained by the THz differential time-domain spectroscopy. The reflection spectra of the GaAs/metals interface were calculated according to the Fresnel formula. The mean reflectance of 25 nm Cr, Ni and Ti are over 80% from 0.3 to 1.5 THz. The results show that ultra-thin metal films can be used for reflectors as well as the electrodes in the THz band.

Appendiceal bleeding: A case report.

BACKGROUND: Acute lower gastrointestinal bleeding is common in clinical practice, and the colon is responsible for the majority of cases. However, appendiceal bleeding is an extremely rare cause. Appendiceal bleeding due to vascular diseases, such as angiodysplasia and Dieulafoy's lesion, may result in massive lower gastrointestinal bleeding. Appendectomy is a reliable and effective option for treatment. CASE SUMMARY: A 32-year-old male presented to our hospital with hematochezia that had lasted for 6 h, with approximately 600-800 mL bloody stools and loss of consciousness for a few seconds. Persistent bleeding from the orifice of the appendix was observed by colonoscopy. Following the new diagnosis of appendiceal bleeding, the patient was treated by an emergency laparoscopic appendectomy. Finally, the patient was pathologically diagnosed with appendiceal Dieulafoy's lesion. The patient was uneventfully discharged, and follow-up 2 wk later showed no evidence of rebleeding. CONCLUSION: Although appendiceal bleeding is a rare cause of acute lower gastrointestinal bleeding, clinicians should consider it during differential diagnosis.

Impact of socio-economic environment and its interaction on the initial spread of COVID-19 in mainland China.

Coronavirus disease 2019 (COVID-19) has strongly impacted society since it was first reported in mainland China in December 2020. Understanding its spread and consequence is crucial to pandemic control, yet difficult to achieve because we deal with a complex context of social environment and variable human behaviour. However, few efforts have been made to comprehensively analyse the socio-economic influences on viral spread and how it promotes the infection numbers in a region. Here we investigated the effect of socio-economic factors and found a strong linear relationship between the gross domestic product (GDP) and the cumulative number of confirmed COVID-19 cases with a high value of R2 (between 0.57 and 0.88). Structural equation models were constructed to further analyse the social-economic interaction mechanism of the spread of COVID-19. The results show that the total effect of GDP (0.87) on viral spread exceeds that of population influx (0.58) in the central cities of mainland China and that the spread mainly occurred through its interplay with other factors, such as socio-economic development. This evidence can be generalized as socio-economic factors can accelerate the spread of any infectious disease in a megacity environment. Thus, the world is in urgent need of a new plan to prepare for current and future pandemics.

Effectiveness and Safety of Acupoint Catgut Embedding for the Treatment of Poststroke Constipation: A Systematic Review and Meta-Analysis.

Objectives: Acupoint catgut embedding therapy has shown effectiveness in treating functional constipation; however, relevant, high-quality clinical evidence is scarce. This study aimed to systematically assess the effectiveness and safety of acupoint catgut embedding in treating poststroke constipation. Methods: Correlative randomized controlled trials were identified through a comprehensive literature search of PubMed, Cochrane Library/Cochrane Central Register of Controlled Trials, Web of Science, Embase, China National Knowledge Internet, Chinese Biomedical Literature Database, Wanfang, and VIP databases from inception until February 2022. Meta-analysis was performed using RevMan 5.3 software. Results: Fifteen trials involving 1084 patients were identified. The meta-analysis revealed that the acupoint catgut embedding group was significantly superior to the non-catgut embedding group with regard to the efficacy rate (RR = 1.27, 95% CI (1.19, 1.37), P < 0.05), the first defecation time (MD = -3.08, 95% CI (-4.53, -1.63), P < 0.05), the defecation sensation score (MD = -0.44, 95% CI (-0.61, -0.26), P < 0.05), the degree of difficulty in defecation (MD = -0.73, 95% CI (-1.10, -0.37), P < 0.05), the PAC-QOL scale score (MD = -10.06, 95% CI (-13.47, -6.64), P < 0.05), and the symptom integral (MD = -3.15, 95% CI (-3.60, -2.71), P < 0.05). However, there was no significant difference in the stool property score (MD = 0.06, 95% CI (-0.39, 0.50), P > 0.05) as well as the incidence of adverse reactions (RD = 0.01, 95% CI (-0.01, 0.03), P > 0.05) between the two groups. Conclusions: The results showed that acupoint catgut embedding is probably an effective and safe acupuncture treatment strategy for poststroke constipation. Nevertheless, more rigorously designed, standardized, large-sample, and multicenter randomized controlled designs are warranted to further verify the findings of this study.

Direct and indirect impacts of urbanization on vegetation growth across the world's cities.

Urban environments, regarded as "harbingers" of future global change, may exert positive or negative impacts on urban vegetation growth. Because of limited ground-based experiments, the responses of vegetation to urbanization and its associated controlling factors at the global scale remain poorly understood. Here, we use satellite observations from 2001 to 2018 to quantify direct and indirect impacts of urbanization on vegetation growth in 672 worldwide cities. After controlling for the negative direct impact of urbanization on vegetation growth, we find a widespread positive indirect effect that has been increasing over time. These indirect effects depend on urban development intensity, population density, and background climate, with more pronounced positive effects in cities with cold and arid environments. We further show that vegetation responses to urbanization are modulated by a cities' developmental status. Our findings have important implications for understanding urbanization-induced impacts on vegetation and future sustainable urban development.

Low-grade elevation of palmitate and lipopolysaccharide synergistically induced ß-cell damage via inhibition of neutral ceramidase.

High concentrations of free fatty acids (FFAs) or lipopolysaccharide (LPS) could lead to ß-cell apoptosis and dysfunction, while low-grade elevation of FFAs or LPS, which are more common in people with type 2 diabetes mellitus (T2DM) or obesity, have no obvious toxic effect on ß-cells. Palmitate is a component closely related to metabolic disorders in FFAs. Recent studies have found that low-grade elevation of palmitate and LPS synergistically affects the sphingolipid signaling pathway by activating Toll-like receptor 4 (TLR4) and further enhances the expression of inflammatory cytokines in immune cells. Previous studies demonstrated that sphingolipids also played an important role in the occurrence and development of T2DM. This study aimed to investigate the synergistic effects of low-grade elevation of palmitate and LPS on viability, apoptosis and insulin secretion in the rat pancreatic ß-cell line INS-1 or islets and the role of sphingolipids in this process. We showed that low-grade elevation of palmitate or LPS alone did not affect the viability, apoptosis, glucose-stimulated insulin secretion (GSIS) or intracellular insulin content of INS-1 cells or islets, while the combination of the two synergistically inhibited cell viability, induced apoptosis and decreased basal insulin secretion in INS-1 cells or islets. Treatment with palmitate and LPS markedly upregulated TLR4 protein expression and downregulated neutral ceramidase (NCDase) activity and protein expression. Additionally, low-grade elevation of palmitate and LPS synergistically induced a significant increase in ceramide and a decrease in sphingosine-1-phosphate. Blocking TLR4 signaling or overexpressing NCDase remarkably attenuated INS-1 cell injury induced by the combination of palmitate and LPS. However, inhibition of ceramide synthase did not ameliorate injury induced by palmitate and LPS. Overall, we show for the first time that low-grade elevation of palmitate and LPS synergistically induced ß-cell damage by activating TLR4 signaling, inhibiting NCDase activity, and further modulating sphingolipid metabolism, which was different from a high concentration of palmitate-induced ß-cell injury by promoting ceramide synthesis.

Mechanism of pod shattering in the forage legume Medicago ruthenica.

Pod shattering is a seed dispersal strategy and an important agronomical trait in domesticated crops. The relationship between pod shattering and pod morphology in the genus Medicago is well known; however, the detailed mechanism underlying pod dehiscence in Medicago ruthenica, a perennial legume used for forage production, is unknown. Here, the pod ventral sutures of shatter-resistant and shatter-susceptible M. ruthenica genotypes were examined at 8, 12, 16, and 20 d after flowering. The mechanism of pod shattering was analyzed through microscopic observations, polygalacturonase (PG) and cellulase (CE) activity analyses, and RNA-sequencing (RNA-Seq), and the results were verified via reverse transcriptase-quantitative polymerase chain reaction. Pod shattering at the ventral suture in M. ruthenica occurs via a combination of two mechanisms: degradation of the middle lamella at the abscission layers (ALs) and detachment of lignified cells on either side of the ALs triggered by physical forces. Increased PG and CE activities in the pod ventral suture are essential for AL cell-autolysis in the shatter-susceptible genotype. RNA-Seq revealed that 11 genes encoding PG and CE were highly expressed in the ventral sutures of the shatter-susceptible genotype. The expression levels of auxin biosynthesis-related genes decreased in the AL cells and they were negatively associated with pod dehiscence. These results enhance our understanding of the pod shattering mechanism not only in M. ruthenica but also in other leguminous plants.