Progress on regulatory elements of plant plastid genetic engineering.

With the advancement of plant synthetic biology, plastids have emerged as an optimal platform for the heterologous production of numerous commercially valuable secondary metabolites and therapeutic proteins. In comparison on nuclear genetic engineering, plastid genetic engineering offers unique advantages in terms of efficient expression of foreign genes and biological safety. However, the constitutive expression of foreign genes in the plastid system may impede plant growth. Therefore, it is imperative to further elucidate and design regulatory elements that can achieve precise regulation of foreign genes. In this review, we summarize the progress made in developing regulatory elements for plastid genetic engineering, including operon design and optimization, multi-gene coexpression regulation strategies, and identification of new expression regulatory elements. These findings provide valuable insights for future research.

Correlation of the spin state and catalytic property of M-N4 single-atom catalysts in oxygen reduction reactions.

The rational design of high-performance catalysts for oxygen reduction reactions (ORRs) is of great importance for large-scale applications in the field of proton-exchange membrane fuel cells and the green synthesis of H2O2. The effect of spin states of paramagnetic metal ions on the selectivity of ORRs is significant for single-atom catalysts (SACs). In this work, via spin-polarization density functional theory (DFT) calculations, we systematically investigated the popular paramagnetic metal-nitrogen graphene (M-N4-C, M = Mn, Fe, and Co) SACs to mainly focus on the correlation of spin states and catalytic performance (e.g. activity and selectivity). Both thermodynamically and kinetically, it was found that Co-N4-C (S = 1/2) has excellent 2e- oxygen reduction performance (hydrogen peroxide production) with an ultralow overpotential of 0.03 V, and the hydrogenation of OOH* is the rate-determining step (RDS) with an energy barrier of 1.20 eV. The 4e- ORR tends to occur along the OOH dissociation pathway (O* + OH*) on Co-N4-C (S = 3/2), in which OOH* decomposition is the RDS with an energy barrier of 1.01 eV. It is proved that the spin magnetic moment is the key factor to regulate the ORR property via multi-angle electronic analysis. The spin states of catalysts play a crucial role in the activity and selectivity of ORRs mainly by manipulating the bond strength between OOH and catalysts. This will provide new insights for the rational design of ORR catalysts with magnetic metals.

Comparative transcriptome and metabolome analyses reveal the methanol dissimilation pathway of Pichia pastoris.

BACKGROUND: Pichia pastoris (Komagataella phaffii) is a model organism widely used for the recombinant expression of eukaryotic proteins, and it can metabolize methanol as its sole carbon and energy source. Methanol is oxidized to formaldehyde by alcohol oxidase (AOX). In the dissimilation pathway, formaldehyde is oxidized to CO2 by formaldehyde dehydrogenase (FLD), S-hydroxymethyl glutathione hydrolase (FGH) and formate dehydrogenase (FDH). RESULTS: The transcriptome and metabolome of P. pastoris were determined under methanol cultivation when its dissimilation pathway cut off. Firstly, Δfld and Δfgh were significantly different compared to the wild type (GS115), with a 60.98% and 23.66% reduction in biomass, respectively. The differential metabolites between GS115 and Δfld were mainly enriched in ABC transporters, amino acid biosynthesis, and protein digestion and absorption. Secondly, comparative transcriptome between knockout and wild type strains showed that oxidative phosphorylation, glycolysis and the TCA cycle were downregulated, while alcohol metabolism, proteasomes, autophagy and peroxisomes were upregulated. Interestingly, the down-regulation of the oxidative phosphorylation pathway was positively correlated with the gene order of dissimilation pathway knockdown. In addition, there were significant differences in amino acid metabolism and glutathione redox cycling that raised our concerns about formaldehyde sorption in cells. CONCLUSIONS: This is the first time that integrity of dissimilation pathway analysis based on transcriptomics and metabolomics was carried out in Pichia pastoris. The blockage of dissimilation pathway significantly down-regulates the level of oxidative phosphorylation and weakens the methanol assimilation pathway to the point where deficiencies in energy supply and carbon fixation result in inefficient biomass accumulation and genetic replication. In addition, transcriptional upregulation of the proteasome and autophagy may be a stress response to resolve formaldehyde-induced DNA-protein crosslinking.

Cooperatively interface role of surface atoms and aqueous media on single atom catalytic property for H2O2 synthesis.

Direct electrosynthesis of hydrogen peroxide (H2O2) from H2 and O2 is a promising alternative to currently industrial Riedl-Pfleiderer route. Utilizing a combination of density functional theory (DFT) and ab-initio-molecular dynamic simulation (AIMD), we presented an effective computational framework to identify the cooperative role of surface atoms(e.g. O, N and S) and aqueous media on catalytic performance of single-atom catalysts (SACs) supported Nb2C MXenes. Computational results shown that both Ni/Nb2CN2 and Co/Nb2CS2 have low overpotentials of 0.17 V and 0.20 V, and the barrier of 0.89 eV and 0.67 eV for 2e- ORR under gas phase, respectively, while in aqueous phase, hydrogen bond framework on the surface promotes the transfer of proton, resulting in the lower 2e- ORR overpotential (0.05 V) in Co/Nb2CS2 and lower barrier (almost 0.01 eV) for rate-determining step (RDS) in Ni/Nb2CN2. Electronically, we found that the less-electronegativity N and S relative to O more benefit to mediate the activation degree of O2 on SACs and thereby improve catalytic selectivity. Thus, it is concluded that both surface atom and aqueous medium synergistically promote catalytic property for H2O2 synthesis.

A new highly oxygenated germacranolide from Carpesium nepalense var. lanatum (C.B.Clarke) Kitam.

A new highly oxygenated germacranolide, carcerlane A (1), together with four known highly oxygenated germacranolides (2-5), was isolated from an ethanol extract of the whole plant of Carpesium nepalense var. lanatum (C.B.Clarke) Kitam. The structures were determined by HRESIMS and extensive analysis of their spectroscopic data including IR, 1 D and 2 D NMR spectra. To our best knowledge, it was the first time to report the phytochemical investigation on this plant. The anti-Alzheimer's disease (AD) activities of 1-5 were evaluated using Caenorhabditis elegans AD pathological model. All the tested compounds showed that they have the anti-AD bioactivities of delaying worms paralysis.

miR-4443 targets TRIM14 to suppress metastasis and energy metabolism of papillary thyroid carcinoma (PTC) in vitro.

Tripartite motif-containing protein 14 (TRIM14) is a tumor-promoter in papillary thyroid carcinoma (PTC). We found that miR-4443 expression was significantly downregulated in PTC tumor tissue, and was negatively associated with TRIM14. This study was designed to investigate the relationship between miR-4443 and TRIM14 on metastasis and energy metabolism in PTC and the underlying mechanisms. To this end, human PTC cells (SW1736 and MZ-CRC-1) were transfected with a miR-4443 mimic or miR-4443 inhibitor + siRNA-TRIM14, and then dual-luciferase assay, Transwell, Seahorse, and western blot analyses were performed to assess the function of miR-4443 and the underlying mechanism. We found that ectopic expression of miR-4443 inhibited PTC cell migration, invasion, ATP production, and aerobic glycolysis, while inhibition of miR-4443 had the opposite effect. miR-4443 directly targeted TRIM14 and reduced both TRIM14 mRNA and protein levels. Silencing TRIM14 significantly reversed miR-4443 inhibition-induced PTC cell migration, invasion, ATP production, aerobic glycolysis, and phosphorylation of the transcription factor STAT3. These findings suggest that miR-4443 is a tumor suppressor in PTC and inhibits metastasis and energy metabolism via the suppression of TRIM14 signaling.

Value of perfusion parameters and histogram analysis of triphasic computed tomography in pre-operative prediction of histological grade of hepatocellular carcinoma.

BACKGROUND: Pre-operative non-invasive histological evaluation of hepatocellular carcinoma (HCC) remains a challenge. Tumor perfusion is significantly associated with the development and aggressiveness of HCC. The purpose of the study was to evaluate the clinical value of quantitative liver perfusion parameters and corresponding histogram parameters derived from traditional triphasic enhanced computed tomography (CT) scans in predicting histological grade of HCC. METHODS: Totally, 52 patients with HCC were enrolled in this retrospective study and underwent triple-phase enhanced CT imaging. The blood perfusion parameters were derived from triple-phase CT scans. The relationship of liver perfusion parameters and corresponding histogram parameters with the histological grade of HCC was analyzed. Receiver operating characteristic (ROC) curve analysis was used to determine the optimal ability of the parameters to predict the tumor histological grade. RESULTS: The variance of arterial enhancement fraction (AEF) was significantly higher in HCCs without poorly differentiated components (NP-HCCs) than in HCCs with poorly differentiated components (P-HCCs). The difference in hepatic blood flow (HF) between total tumor and total liver flow (ΔHF = HFtumor - HFliver) and relative flow (rHF = ΔHF/HFliver) were significantly higher in NP-HCCs than in P-HCCs. The difference in portal vein blood supply perfusion (PVP) between tumor and liver tissue (ΔPVP) and the ΔPVP/liver PVP ratio (rPVP) were significantly higher in patients with NP-HCCs than in patients with P-HCCs. The area under ROC (AUC) of ΔPVP and rPVP were both 0.697 with a high sensitivity of 84.2% and specificity of only 56.2%. The ΔHF and rHF had a higher specificity of 87.5% with an AUC of 0.681 and 0.673, respectively. The combination of rHF and rPVP showed the highest AUC of 0.732 with a sensitivity of 57.9% and specificity of 93.8%. The combined parameter of ΔHF and rPVP, rHF and rPVP had the highest positive predictive value of 0.903, and that of rPVP and ΔPVP had the highest negative predictive value of 0.781. CONCLUSION: Liver perfusion parameters and corresponding histogram parameters (including ΔHF, rHF, ΔPVP, rPVP, and AEFvariance) in patients with HCC derived from traditional triphasic CT scans may be helpful to non-invasively and pre-operatively predict the degree of the differentiation of HCC.

Isolation, Characterization, and Possible Anti-Alzheimer's Disease Activities of Bisabolane-Type Sesquiterpenoid Derivatives and Phenolics from the Rhizomes of Curcuma longa.

One new bisabolane-type sesquiterpenoid, together with four known bisabolane-type sesquiterpenoid derivatives and seven phenolics, was isolated from the rhizomes of Curcuma longa. Their structures were elucidated by extensive spectroscopic (IR, HR-ESI-MS, and NMR) data analysis. The possible anti-Alzheimer's disease (AD) activities of the isolated compounds were also evaluated using Caenorhabditis elegans AD pathological model, and 1ß-hydroxybisabola-2,10-dien-4-one had the highest possible anti-AD activity.

Cadinane-type sesquiterpenes from the resinous exudates of Commiphora myrrha and their anti-Alzheimer's disease bioactivities.

Commiphoins A-C (1-3), three new cadinane-type sesquiterpenes, together with two known cadinane-type sesquiterpenes (4 and 5) were isolated from the resinous exudates of Commiphora myrrha. Their structures and relative configurations were established on the basis of comprehensive spectroscopic methods, including HRESIMS, 1D and 2D NMR analyses. Compounds 1 and 3-5 were screened for anti-Alzheimer's disease (AD) activities using the AD pathological model in Caenorhabditis elegans. The results showed that they all had significant anti-AD activities.

Eupulcherol A, a triterpenoid with a new carbon skeleton from Euphorbia pulcherrima, and its anti-Alzheimer's disease bioactivity.

Eupulcherol A (1), a novel triterpenoid with an unprecedented carbon skeleton, was isolated from Euphorbia pulcherrima. Its structure was determined by comprehensive analysis of spectroscopic data, including HRESIMS and 1D and 2D NMR, and the absolute configuration was defined by single crystal X-ray diffraction analysis. Biological studies showed that compound 1 possessed anti-Alzheimer's disease (AD) bioactivity, which could delay paralysis of transgenic AD Caenorhabditis elegans. A plausible biogenetic pathway for eupulcherol A (1) was also proposed.

Narjatamanins A and B, a pair of novel epimers possessing a 2,3-seco-iridoid skeleton with an unusual 1,10-oxygen bridge from Nardostachys jatamansi and evaluation of their effects on worm paralysis in AD C. elegans.

Narjatamanins A (1) and B (2), a pair of epimers possessing a novel 2,3-seco-iridoid skeleton with an unusual 1,10-oxygen bridge, were isolated from the whole plants of Nardostachys jatamansi. Their structures were elucidated by a combination of various spectroscopic methods, including HRESIMS, IR and 1D and 2D NMR techniques. The absolute configurations of 1 and 2 were established by electronic circular dichroism (ECD) calculations. The pharmacological activities of 1 and 2 to alleviate AD-like symptoms were also evaluated using the Caenorhabditis elegans Alzheimer's disease (AD) pathological model, and narjatamanins A (1) and B (2) showed statistically significant delay in the worm paralysis phenotype of AD-like symptoms.

Four novel sesquiterpenoids with their anti-Alzheimer's disease activity from Nardostachys chinensis.

Nardochinins A-D (1-4), four novel sesquiterpenoids, along with four known ones were isolated from the underground parts of Nardostachys chinensis Batal in ethanol. Their structures were determined by extensive spectroscopic methods and single-crystal X-ray diffraction. Nardochinin A (1) possessed a norsesquiterpene skeleton with an unusual 3/6/5/5 tetracyclic ring system, which had not appeared in natural products. Nardochinins B (2) and C (3) were the first time found naturally occurring sesquiterpenoids with a 4,5-seco-nardosinane skeleton. Besides, compound 3 represented an unprecedented 4,5-seco-nardosinane type norsesquiterpenoid with losing an isopropenyl at C-6 compared with 2 in the structural framework. Nardochinin D (4) was a novel, highly oxygenated valerenane-type sesquiterpenoid possessing a rare 3,12-epoxy group and an unusual 9,11-epoxy group. The anti-Alzheimer's disease (AD) activities of 1-4 were also evaluated using the Caenorhabditis elegans AD pathological model, and nardochinin B (2) had the highest anti-AD activity.

The feasibility of 1-stop examination of coronary CT angiography and abdominal enhanced CT.

This study aims to evaluate the feasibility of performing coronary computed tomography angiography (CCTA) and abdominal enhanced computed tomography (CT) with 1-time injection of the agent.CCTA images (right coronary artery, left anterior descending coronary artery, and left circumflex coronary artery) were collected from 20 patients who completed a 1-stop combined examination of CCTA and abdominal enhanced CT (group A), 20 patients who only underwent abdominal enhanced CT (group B1), and 20 patients who only underwent CCTA (group B2). These images were interpreted using the 5-point Likert scale system by 2 experienced radiologists, and abdominal images were observed for breathing artifact. CT value, signal-to-noise ratio (SNR), and CTDI were recorded and compare among the 3 groups.The difference in image quality of the coronary and total volume of the contrast agent between group A and group B1 was not statistical significant (P > .05). The CT value and SNR in group B1 (CCTA) (CT: 394.65 ±â€Š59.23, SNR: 17.38 ±â€Š4.13) increased, compare with Group A (CT: 360.35 ±â€Š34.16, SNR: 13.76 ±â€Š1.84, P = .03, .01), while CTDI was undifferentiated between group A (17.14 ±â€Š6.20) and group B1 (18.38 ±â€Š9.79) (P = .64). The difference in CT value and SNR at the arterial phase and CT value at the venous phase between group A (abdomen) and group B2 were statistically significant, the CTDI in group A (9.09 ±â€Š1.05) increased, compared with group B2 (8.23 ±â€Š1.33) (P = .03), and SNR at the venous phase in group B2 (12.50 ±â€Š2.43) increased, compared with group A (10.89 ±â€Š2.03) (P = .03).Revolution CT can capture full images and very rapidly switch to the scan mode, enabling a 1-stop axial CCTA and enhanced helical abdominal scan. The 1-stop combined scan resulted in a satisfactory image quality, which reduced the contrast agent dose and simplified the workflow.The 1-stop combined scan allows for the high success rate of the examination, reduces the number of examinations, and decreases the dose and risk of injection of the contrast agent. This would be helpful for patients to obtain diagnostic images in time.

[Establishment of a genetic transformation system for hairy root culture of Erigeron breviscapus].

The electroporation method was performed to transfer plasmid DNA of PBI-1300 carrying GFP gene into Agrobacterium rhizogenes C58C1 strains. Mediated by A. rhizogenes C58C1, the GFP gene were transformed into Erigeron breviscapus aseptic leaves by leaf disc method, then the hairy roots were induced and the infected hairy roots were screened by hygromycin resistance. The chromosomal DNA of the hairy root was used as the templates for the PCR amplification with the GFP-specific primers and then the expected amplified DNA bands appeared, the green fluorescent of GFP in the cut hairy roots was observed by two-photon microscope. These results indicated that GFP gene was integrated into the genome of E. breviscapus and was expressed stably. This study laid the groundwork for foreign gene high-efficiency expression inthe genetic transformation system for hairy root culture of E. breviscapus.

Application of 3D printing rapid prototyping-assisted percutaneous fixation in the treatment of intertrochanteric fracture.

The aim of the present study was to investigate the application of 3D printing (3DP) rapid prototyping (RP) technique-assisted percutaneous fixation in the treatment of femoral intertrochanteric fracture (ITF) using proximal femoral nail anti-rotation (PFNA). A total of 39 patients with unstable ITF were included in the current study. Patients were divided into two groups: 19 patients were examined using computed tomography scanning and underwent PFNA with SDP-RP whereas the other 20 patients underwent conventional PFNA treatment. Anatomical data were converted from the Digital Imaging and Communications in Medicine format to the stereolithography format using M3D software. The 3DP-RP model was established using the fused deposition modeling technique and the length and diameter of the main screw blade was measured during the simulation. The postoperative femoral neck-shaft angle (NSA), surgery duration, intraoperative and postoperative blood loss, and the duration of hospital stay were recorded and compared with the corresponding values in conventional surgery. No significant differences were observed in mean PFNA size between the implants used and the preoperative planning estimates. It was demonstrated that the 3DP-RP assisted procedure resulted in more effective reduction of the NSA. Furthermore, patients undergoing 3DP-RP experienced a significant reduction in duration of surgery (P<0.01), as well as reductions in intraoperative (P=0.02) and postoperative (P=0.03) blood loss, compared with conventional surgery. At 6 months post-surgery, no cases of hip varus/vague deformities or implant failure were observed in patients that underwent either the 3DP-RP-assisted or conventional procedure. The results of the present study suggest that the 3DP-RP technique is able to create an accurate model of the ITF, which facilitates surgical planning and fracture reduction, thus improving the efficiency of PFNA surgery for ITFs.

[Habitat suitability and quality division of Mentha haplocalyx].

In this study, ecological factors, occurrence records, the essential oil components content were used to predict the potential geographical distribution and quality division of Mentha haplocalyx in China based on the MaxEnt modeling and geographic information system(GIS). The AUC of ROC curve was above 0.950,indicating that the predictive results with the maximum model were highly precise. The results showed that the main environmental factors determining the potential distribution were annual average precipitation (the contribution rate, 45.87%), mean temperature of wettest quarter (11.92%), mean temperature of warmest quarter (7.84%), average monthly precipitation of May (6.80%), standard deviation of seasonal temperature variation (4.42%), mean temperature of the coldest quarter (3.47%) and altitude (2.92%). The environmental variables in the highly potential areas were determined as annual average precipitation around [530,1 465] mm, mean temperature of wettest quarter around [24.5,29] ℃, mean temperature of the warmest quarter around [25.5,29] ℃, average monthly precipitation of May around [67,133] mm, standard deviation of temperature seasonal change around [8 333,9 643], mean temperature of the coldest quarter around [1.7,8.3] ℃ and the altitude around [0,165] mm. The best quality distribution of M. haplocalyx was mainly located in Jiangsu, Anhui, Shandong, Zhejiang and Heilongjiang. The zoning results basically coincide with the actual situation. The quality division of M. haplocalyx can be used for providing a scientific basis for selection of artificial planting base and guidance of its production.