Chromosome-level Alstonia scholaris genome unveils evolutionary insights into biosynthesis of monoterpenoid indole alkaloids.

Alstonia scholaris of the Apocynaceae family is a medicinal plant with a rich source of bioactive monoterpenoid indole alkaloids (MIAs), which possess anti-cancer activity like vinca alkaloids. To gain genomic insights into MIA biosynthesis, we assembled a high-quality chromosome-level genome for A. scholaris using nanopore and Hi-C data. The 444.95 Mb genome contained 35,488 protein-coding genes. A total of 20 chromosomes were assembled with a scaffold N50 of 21.75 Mb. The genome contained a cluster of strictosidine synthases and tryptophan decarboxylases with synteny to other species and a saccharide-terpene cluster involved in the monoterpenoid biosynthesis pathway of the MIA upstream pathway. The multi-omics data of A. scholaris provide a valuable resource for understanding the evolutionary origins of MIAs and for discovering biosynthetic pathways and synthetic biology efforts for producing pharmaceutically useful alkaloids.

Highly selective photoelectrochemical CO2 reduction by crystal phase-modulated nanocrystals without parasitic absorption.

Photoelectrochemical (PEC) carbon dioxide (CO2) reduction (CO2R) holds the potential to reduce the costs of solar fuel production by integrating CO2 utilization and light harvesting within one integrated device. However, the CO2R selectivity on the photocathode is limited by the lack of catalytic active sites and competition with the hydrogen evolution reaction. On the other hand, serious parasitic light absorption occurs on the front-side-illuminated photocathode due to the poor light transmittance of CO2R cocatalyst films, resulting in extremely low photocurrent density at the CO2R equilibrium potential. This paper describes the design and fabrication of a photocathode consisting of crystal phase-modulated Ag nanocrystal cocatalysts integrated on illumination-reaction decoupled heterojunction silicon (Si) substrate for the selective and efficient conversion of CO2. Ag nanocrystals containing unconventional hexagonal close-packed phases accelerate the charge transfer process in CO2R reaction, exhibiting excellent catalytic performance. Heterojunction Si substrate decouples light absorption from the CO2R catalyst layer, preventing the parasitic light absorption. The obtained photocathode exhibits a carbon monoxide (CO) Faradaic efficiency (FE) higher than 90% in a wide potential range, with the maximum FE reaching up to 97.4% at -0.2 V vs. reversible hydrogen electrode. At the CO2/CO equilibrium potential, a CO partial photocurrent density of -2.7 mA cm-2 with a CO FE of 96.5% is achieved in 0.1 M KHCO3 electrolyte on this photocathode, surpassing the expensive benchmark Au-based PEC CO2R system.

Parameter Optimization of Ultrasonic-Microwave Synergistic Extraction of Taxanes from Taxus cuspidata Needles.

Taxanes are the best-known compounds in Taxus cuspidata owing to their strong anticancer effects. However, the traditional taxanes extraction method is the solid-liquid extraction method, which is limited by a large energy consumption and low yield. Therefore, it is urgent to find an efficient method for taxanes extraction. The ultrasonic microwave synergistic extraction (UME) method integrates the cavitation effect of ultrasound and the intensifying heat transfer (ionic conduction and dipole rotation of molecules) effect of microwave to accelerate the release of intracellular compounds and is used in active ingredient extractions. This study aimed to evaluate the performance of UME in extracting taxanes from T. cuspidata needles (dichloromethane-ethanol as extractant). A single-factor experiment, Plackett-Burman design, and the response surface method showed that the optimal UME parameters for taxanes extraction were an ultrasonic power of 300 W, a microwave power of 215 W, and 130 sieve meshes. Under these conditions, the taxanes yield was 570.32 µg/g, which increased by 13.41% and 41.63% compared with the ultrasound (US) and microwave (MW) treatments, respectively. The reasons for the differences in the taxanes yield were revealed by comparing the physicochemical properties of T. cuspidata residues after the UME, US, and MW treatments. The cell structures were significantly damaged after the UME treatment, and numerous tiny holes were observed on the surface. The absorption peaks of cellulose, hemicellulose, and lignin increased significantly in intensity, and the lowest peak temperature (307.40 °C), with a melting enthalpy of -5.19 J/g, was found after the UME treatment compared with the US and MW treatments. These results demonstrate that UME is an effective method (570.32 µg/g) to extract taxanes from T. cuspidata needles by destroying cellular structures.

The draft genome and multi-omics analyses reveal new insights into geo-herbalism properties of Citrus grandis 'Tomentosa'.

Citrus grandis 'Tomentosa' (CGT) (Huajuhong, HJH) is a widely used medicinal plant, which is mainly produced in Guangdong and Guangxi provinces of South China. Particularly, HJH from Huazhou (HZ) county of Guangdong province has been well-regarded as the best national product for geo-herbalism. But the reasons for geo-herbalism property in HJH from HZ county remains a mystery. Therefore, a multi-omics approach was applied to identify the nature of the geo-herbalism in CGT from three different regions. The comprehensive screening of differential metabolites revealed that the Nobiletin content was significantly different in HZ region compared to other regions, and could be employed as a key indicator to determine the geo-herbalism. Furthermore, the high-quality genome (N50 of 9.12 Mb), coupled with genomics and transcriptomics analyses indicated that CGT and Citrus grandis are closely related, with a predicted divergence time of 19.1 million years ago (MYA), and no recent WGD occurred in the CGT, and the bioactive ingredients of CGT were more abundant than that of Citrus grandis. Interestingly, Nobiletin (Polymethoxyflavones) content was identified as a potential indicator of geo-herbalism, and O-methyltransferase (OMT) genes are involved in the synthesis of Polymethoxyflavones. Further multi-omics analysis led to the identification of a novel OMT gene (CtgOMT1) whose transient overexpression displayed significantly higher Nobiletin content, suggesting that CtgOMT1 was involved in the synthesis of Nobiletin. Overall, our findings provide new data resources for geo-herbalism evaluation, germplasm conservation and insights into Nobiletin biosynthesis pathways for the medicinal plant C. grandis 'Tomentosa'.

An injectable thermosensitive hydrogel loaded with a theranostic nanoprobe for synergistic chemo-photothermal therapy for multidrug-resistant hepatocellular carcinoma.

Multi-drug resistance (MDR) is a complicated cellular defense mechanism for tumor cells to resist chemotherapy drugs, which is also the main cause of chemotherapy failure. In this study, a local injectable hydrogel delivery system was used to construct an on-demand sustained-release platform with the advantages of chemotherapy, photothermal therapy (PTT), and magnetic resonance imaging (MRI). It could achieve synergistic chemo-photothermal therapy and real-time evaluation of the therapeutic effects (via MRI) for MDR hepatocellular carcinoma (HCC). Furthermore, after a single administration, the prepared hydrogel with a theranostic nanoprobe could release the therapeutic agents on demand for up to 14 d. Firstly, doxorubicin (DOX) and gold-manganese oxide (Au-MnO) nanoparticles (NPs) were incorporated into liposome-based self-assembled micelles, then loaded into the thermosensitive hydrogel (F127) to form DOX@Au-MnO-L NPs/F127 hydrogel (DAML/H). The prepared NP complex showed a spherical morphology with a narrow size distribution. The prepared hydrogel drug delivery system had injectable properties and stable photothermal conversion. Both the DOX@Au-MnO-L NPs and DAML/H showed controlled drug release under near infrared (NIR) laser irradiation. The in vitro MRI studies indicated that the prepared DAML/H had a high relaxation rate (14.38 mM-1 s-1) and good MRI scanning sensitivity conditions. The in vitro and in vivo results suggested the synergistic chemo-photothermal therapy of DAML/H with NIR irradiation (808 nm, 1 W cm-2, 10 min) improved the antitumor efficacy for MDR HCC. The in vivo retention experiment of Au in tumors indicated that the prepared hydrogel drug delivery system (DAML/H) had a good ability to retain Au in the tumor for a long time (at least 14 d). The western blotting results revealed that DAML/H with laser treatment could effectively downregulate P-glycoprotein (P-gp), p53 and antiapoptotic protein (Bcl-2), whereas the expression level of proapoptotic protein (Bax) and caspase-3 were increased. Therefore, DAML/H could serve as a promising synergistic chemo-photothermal therapy for MDR HCC, and a single administration might achieve long-term (14 d), on-demand, sustained-release treatment of tumors.

Severe complications and management of a patient with myasthenia gravis undergoing anterior cervical spinal surgery: a case report.

Surgical management of patients with comorbid long-term myasthenia gravis (MG) is particularly challenging and MG thus represents an independent risk factor for perioperative complications. However, few studies have reported on the perioperative assessment, prevention measures, and risks in MG patients undergoing major surgery, especially for anterior cervical spine surgery. We herein report the rare case of a 62-year-old man with a 20-year history of MG, who was admitted to our hospital with diagnosis of degenerative cervical spondylosis. He safely underwent anterior cervical corpectomy of C4, discectomy of C5-6, and fusion of C3-6. Intraoperative motor evoked potential was recorded to detect significant improvement after decompression. However, the patient suffered from progressive dysphagia, bucking, and hyperpyrexia 20 days after the initial operation. Imaging revealed titanium cage sliding and graft dislodgement. Secondary surgery was performed for posterior internal fixation from C2-7 and anterior revision from C3-6 after Halo-Vest traction, antibiotic treatment, and immunoglobulin therapy. He underwent a series of postoperative treatments, including cervicothoracolumbosacral orthosis, atomization inhalation, chest physiotherapy, antibiotics, and nutritional support. His condition improved markedly and he had no recurrence of symptoms during the 6-month follow-up. It is the rare reported case of anterior cervical spinal surgery in a patient with MG. This rare case indicates a relative contraindication to anterior-only approaches especially with multiple levels for MG patients with cervical spondylosis. Posterior approach, intraoperative monitoring, osteoporosis, postoperative strong brace protection, and supportive management should be considered for patients who were on large doses of steroids for long duration of time, given the lack of sufficient bone mineral density.

Combined berberine and probiotic treatment as an effective regimen for improving postprandial hyperlipidemia in type 2 diabetes patients: a double blinded placebo controlled randomized study.

Non-fasting lipidemia (nFL), mainly contributed by postprandial lipidemia (PL), has recently been recognized as an important cardiovascular disease (CVD) risk as fasting lipidemia (FL). PL serves as a common feature of dyslipidemia in Type 2 Diabetes (T2D), albeit effective therapies targeting on PL were limited. In this study, we aimed to evaluate whether the therapy combining probiotics (Prob) and berberine (BBR), a proven antidiabetic and hypolipidemic regimen via altering gut microbiome, could effectively reduce PL in T2D and to explore the underlying mechanism. Blood PL (120 min after taking 100 g standard carbohydrate meal) was examined in 365 participants with T2D from the Probiotics and BBR on the Efficacy and Change of Gut Microbiota in Patients with Newly Diagnosed Type 2 Diabetes (PREMOTE study), a random, placebo-controlled, and multicenter clinical trial. Prob+BBR was superior to BBR or Prob alone in improving postprandial total cholesterol (pTC) and low-density lipoprotein cholesterol (pLDLc) levels with decrement of multiple species of postprandial lipidomic metabolites after 3 months follow-up. This effect was linked to the changes of fecal Bifidobacterium breve level responding to BBR alone or Prob+BBR treatment. Four fadD genes encoding long-chain acyl-CoA synthetase were identified in the genome of this B. breve strain, and transcriptionally activated by BBR. In vitro BBR treatment further decreased the concentration of FFA in the culture medium of B. breve compared to vehicle. Thus, the activation of fadD by BBR could enhance FFA import and mobilization in B. breve and diliminish the intraluminal lipids for absorption to mediate the effect of Prob+BBR on PL. Our study confirmed that BBR and Prob (B. breve) could exert a synergistic hypolipidemic effect on PL, acting as a gut lipid sink to achieve better lipidemia and CVD risk control in T2D.

Production of Margarines Rich in Unsaturated Fatty Acids Using Oxidative-stable Vitamin C-Loaded Oleogel.

Vitamin C (VC)-loaded oleogel (VCOG) with corn oil and monoglyceride stearate was used to replace lipid phase of margarine completely. The oxidative stability of VCOG was evaluated at 60±1°C in a lightproof oven for 18 days and the result showed that VCOG peroxide (> 6 days) and p-anisidine value (> 4 days) was significantly lower than that of bulk oil and VC-free oleogel (p < 0.05). Then, the margarine containing 79.70% VCOG (VCOGM) was in comparison with four commercial butter in sensory and physical characteristic. Results showed that firmness, solid fat content and trans fatty acid of VCOGM were in the lowest values while unsaturated fatty acid and adhesiveness of VCOGM was in the highest values. Furthermore, VCOGM presented the similar springiness, cohesiveness, gumminess, score appearance, texture, taste and overall impression to some/all commercial butters selected in this research (p > 0.05). These results implied that VC-loaded oleogel was an excellent alternative of lipid phase in margarine which confirmed by 55% "definitely buy" and 25% "try once-then decide".

Influences of lead exposure on its accumulation in organs, meat, eggs and bone during laying period of hens.

Elevating levels of environmental lead (Pb) results in serious hazards to health of animals and human beings. In this study, daily diet with three different levels of Pb (Pb nitrate at doses of 1, 10, and 100 mg/kg body weight) were fed to ISA Brown layers. It showed that the kidney and liver have the relatively high Pb concentration (2.34 and 0.51 ppm) after culture, while the meat has the Pb concentration as low as 0.07 ppm (lower than the standard of Codex Alimentarius). It was also confirmed that egg laying worked as a potential pathway for hens to excrete Pb as Pb concentrations in eggshell and yolk increased from 0.10 to 3.11 ppm. However, the Pb concentration in egg white remains at a safe level (<0.10 ppm). Furthermore, even the intake of low dose Pb can cause a decline of bone mineral density and bone strength. Raman spot and mapping analysis indicated that carotenoids content in humerus from the hens of high dose group increased significantly, which hence can be applied as an indicator for resist stress. The degradation of bone quality will further damage the health of laying hens. Therefore, Pb exposure not only toxifies organs and reduces physiological features (e.g., body weight and laying rate) instantly, but also hurts poultry via degrading bone quality in long term. Additionally, the probability of excessive Pb in poultry meat is less than those of viscera and eggs, indicating its low risk to food safety.

Chloride-mediated co-formation of 3-monochloropropanediol esters and glycidyl esters in both model vegetable oils and chemical model systems.

3-Monochloropropanediol esters (3-MCPDEs) and glycidyl esters (GEs) with high toxicity have drawn global concerns due to their widespread occurrence in refined oils and oil-based foods. The effect mechanisms of organic chlorine compound lindane, inorganic chlorine compounds tetra-n-butylammonium chloride (TBAC) and sodium chloride (NaCl) on the formation of 3-MCPDEs and GEs were investigated in model oils and chemical models at 240 °C. Results showed that 3-MCPDEs contents increased with the addition of lindane and TBAC, whereas, surprisingly, GEs presented the same tendency as the results of 3-MCPDEs. This suggested that although chlorine compounds were not involved in the formation reaction of GEs, they could also promote GEs formation. Chemical model experiments confirmed that the presence of chlorine compounds led to the transformation of GEs to 3-MCPDEs and conversely 3-MCPDEs could also transform to GEs. The latter transformation rate from 3-MCPDEs to GEs was higher than the former, which might account for the fact that chlorine compounds promoted GEs formation. Additionally, it was also observed that solid NaCl did not induce the increase of 3-MCPDEs and GEs levels in chemical models, suggesting that the chlorine in NaCl, different from lindane and TBAC, was not available for 3-MCPDEs formation. The present findings give novel insights into the interactions between 3-MCPDEs and GEs formation mechanisms, which offer the theoretical basis for efficient and simultaneous inhibition of 3-MCPDEs and GEs.

The inhibitory effects of sesamol and sesamolin on the glycidyl esters formation during deodorization of vegetables oils.

BACKGROUND: Glycidyl esters (GEs) have attracted worldwide attention for their potential harm to human health. The GEs in edible oils mainly form during the deodorization of the oil refining processes. We used sesamol and sesamolin to inhibit the formation of GEs in model corn oil (MCO), model palm oil (MPO) and model rice bran oil (MRO) during a deodorization process. RESULTS: The results showed that, in the three model oils, the total GE content was in the following order from highest to lowest: MRO (1437.98 µg kg-1 ) > MPO (388.64 µg kg-1 ) > MCO (314.81 µg kg-1 ). The inhibitory effect of the three antioxidants on the formation of GEs in the MCO was in the following order from strongest to weakest: tert-butylhydroquinone (TBHQ) > sesamol > sesamolin. CONCLUSION: When the mass percentage of sesamol was 0.05%, its inhibition percentage on GEs was close to the inhibition percentage of 0.02% added TBHQ. The present study provides a foundation for understanding how to inhibit the formation of GEs in oils by adding sesamol during the deodorization process.

Gut microbiome-related effects of berberine and probiotics on type 2 diabetes (the PREMOTE study).

Human gut microbiome is a promising target for managing type 2 diabetes (T2D). Measures altering gut microbiota like oral intake of probiotics or berberine (BBR), a bacteriostatic agent, merit metabolic homoeostasis. We hence conducted a randomized, double-blind, placebo-controlled trial with newly diagnosed T2D patients from 20 centres in China. Four-hundred-nine eligible participants were enroled, randomly assigned (1:1:1:1) and completed a 12-week treatment of either BBR-alone, probiotics+BBR, probiotics-alone, or placebo, after a one-week run-in of gentamycin pretreatment. The changes in glycated haemoglobin, as the primary outcome, in the probiotics+BBR (least-squares mean [95% CI], -1.04[-1.19, -0.89]%) and BBR-alone group (-0.99[-1.16, -0.83]%) were significantly greater than that in the placebo and probiotics-alone groups (-0.59[-0.75, -0.44]%, -0.53[-0.68, -0.37]%, P < 0.001). BBR treatment induced more gastrointestinal side effects. Further metagenomics and metabolomic studies found that the hypoglycaemic effect of BBR is mediated by the inhibition of DCA biotransformation by Ruminococcus bromii. Therefore, our study reports a human microbial related mechanism underlying the antidiabetic effect of BBR on T2D. (Clinicaltrial.gov Identifier: NCT02861261).

The effects of oil type and crystallization temperature on the physical properties of vitamin C-loaded oleogels prepared by an emulsion-templated approach.

Vitamin C (VC) is widely used as an antioxidant and nutrient to increase the nutritional value and shelf-life of foods. In this article, VC was loaded in oleogels using a simple two-step emulsion-templated approach and the effects of oil type (linseed oil, corn oil, and camellia oil) and crystallization temperature (Tc, -18, 0, 5, and 25 °C) on the physical properties, VC concentration, and oxidation stability of the VC-loaded oleogels were studied. As the amount of saturated fatty acids in the oil phase of the oleogels decreased, the VC loading level, oxidation stability and physical properties of the corn-oil-based oleogel (COG) were better than those of camellia-oil-based oleogels and linseed-oil-based oleogels. At different Tc values, the temperature and frequency dependent storage modulus values for the COG crystallized at 0 °C and 5 °C were not significantly different (P > 0.05), but their values were higher than those for COG crystallized at -18 °C and 25 °C (P < 0.05); the firmness of the COG crystallized at -18 °C and 0 °C was higher than those crystallized at 5 °C and 25 °C (P < 0.05); the network of the COG crystallized at 0 °C was denser than those of the COG crystallized at -18 °C, 5 °C, and 25 °C; and the VC concentration of the oleogels was affected by the crystallization temperature (Tc) and temperature fluctuations. To sum up, a VC-loaded oleogel with excellent mechanical properties was prepared using corn oil and crystallized at 0 °C via an emulsion-templated approach.

Delicate changes of bioapatite mineral in pig femur with addition of dietary xylooligosaccharide: Evidences from Raman spectroscopy and ICP.

Bone mineral is strongly correlated with performance and health of animal bodies. The mineral bioapatite (BAp) is the dominant component in bone tissue. This study investigated mineralogical changes of BAp in pig femur by Raman spectroscopy and inductively coupled plasma optical emission spectrometry (ICP-OES). The pigs had been raised with various xylooligosaccharide (XOS) additions at two stages of growth (growing and fattening periods). The results show that XOS can decrease the degree of carbonate substitution for PO4 in BAp mineral and improve the mineral's crystallinity. ICP data is consistent with the Raman results, that is the low solubility of bone BAp for pigs fed with XOS. Additionally, the effect of XOS is much better in the growing period (before 65 kg) than in the fattening period (after 65 kg). Moreover, the high addition of XOS (within the range of 0.1-0.5 g/kg) would be appropriate to improve the crystallinity of bone BAp. This study sheds light on applying Raman and ICP techniques to investigate the delicate changes of mineral in pig bones undergoing different managements.

[Exploring the pathogenesis and therapy of liver cancer from "damp-heat insidious pathogen" to "cancer toxin"].

From a macro-level analysis of the attributes and pathogenic features of HBV, the main pathogenic factor for chronic liver diseases including viral hepatitis, cirrhosis, and liver cancer, the concept of damp-heat insidious pathogen was obtained, according to which, in-depth discussions were undertaken. Adopting syndrome typing of Wei (defense), qi (vital energy), Ying (nutrients), and blood, the pathogens leading to different syndromes as well as new products such as pathological "sputum", "stasis" in the disease process were understood, and then, the pathological "sputum" and "stasis", as the hub, playing a role in chronic lesions of the liver collateral were explained. Finally the pathological "sputum" and "stasis" blend and form cancer toxin. Through a comprehensive understanding of the development of chronic liver diseases, it is clear that damp-heat insidious pathogen, as its initiating factor, always exists in the whole process. We summed up heat clearing, dampness resolving, and detoxification was the principle for treating chronic liver disease.

[Aitongxiao recipe regulated survivin and Bcl- 2 in rats' transplanted hepatoma carcinoma cell].

OBJECTIVE: To study the main mechanisms of Aitongxiao Recipe (ATXR) for anti-tumor at the molecular level, and to clarify different efficient drugs' roles in anti-tumor, thus in-depth explaining the objectivity and substance of "cancer toxic" theory. METHODS: Walker-256 tumor strain was used for Wistar rat transplanted liver cancer modeling. After successful modeling rats were randomly divided into 5 groups, i. e., the ATXP group, the qi regulating and blood circulating group (as the assembled I group), the heat clearing and detoxification group (as the assembled II group), the body resistance strengthening and cultivating group (as the assembled III group), and the model group, 10 in each group. Corresponding medication was given to rats in each group for 14 successive days. Finally rats were sacrificed and the tumor mass was taken out. The apoptosis rate and the cell cycle of tumor cells were detected by flow cytometry Annexin V/PI. The protein and mRNA expressions of Bcl-2 and survivin were detected using immunohistochemistry and real-time fluorescent quantitative PCR. RESULTS: (1) The apoptosis of hepatoma carcinoma cells could be obviously promoted in the ATXP group. The cell cycle could also be affected, making major cells arrest at G0/G1 phase. The proliferation of hepatoma carcinoma cells was effectively prevented. The efficacy in the assembled II group was in line with that in the ATXP group with no statistical difference (P>0.05). It was also effective in the assembled III group, but its efficacy was not as good as that in the former two groups, showing statistical difference (P<0.01). (2) ATXP could obviously down-regulate the protein and mRNA expressions of Bcl-2 and survivin in hepatoma carcinoma cells. Drugs for heat clearing and detoxification showed significant effects on down-regulating the protein and mRNA expressions of Bcl-2 and survivin in hepatoma carcinoma cells. Their effects were similar to that of ATXP (P>0.05). The effects of drugs for body resistance strengthening and cultivating were not as good as the former two, showing statistical difference (P<0.01). Drugs for blood circulating and stasis removing could up-regulate the protein and mRNA expressions of Bcl-2 and survivin to some extent. CONCLUSIONS: (1) ATXP could increase the apoptosis ratio of hepatoma carcinoma cells obviously through down-regulating the protein and mRNA expressions of Bcl-2 and survivin, thus inhibiting their proliferation. (2) Drugs for heat clearing and detoxification played the most important roles in ATXP. The evil heat and dampness (damp-heat insidious pathogen) is the most fundamental carcinogenic factors. The insufficiency of vital qi is also one of the pathogenic factors. The mechanisms of phlegm, stasis, and other pathological products are not clear and await further studies.