Additional hemoperfusion for patients receiving maintenance hemodialysis: a retrospective analysis.

OBJECTIVE: To determine the efficacy of hemodialysis combined with hemoperfusion with acupuncture on calcium-phosphorus metabolism disorder (CPMD) of patients who had received maintenance hemodialysis and its effect on intact parathyroid hormone (iPTH) and nutritional status. METHODS: Data from 142 patients who were treated and given maintenance hemodialysis in Baoji People's Hospital from March 2018 to February 2020 were analyzed retrospectively. Patients treated with hemodialysis and acupuncture-moxibustion adjuvant therapy were enrolled into the control group (n=58), while those treated with hemoperfusion in addition to hemodialysis and acupuncture-moxibustion adjuvant therapy were enrolled into the research group (n=84). The two groups were compared in terms of changes in iPTH, calcium-phosphorus product, serum calcium (Ca), serum phosphorus (P), ß2 microglobulin (ß2-MG), serum albumin (Alb), creatinine (Scr) and urea nitrogen (BUN). The clinical efficacy in the two groups was compared after therapy, and the two groups were also compared in the improvement of immune function-related indexes (IgG and IgM) and the changes of nutrition-related indexes (Alb, prealbumin (PA) and hemoglobin (Hb)) before and after treatment. A risk prediction model was constructed based on LASSO regression to evaluate the predictive value of the risk score for efficacy of patients. RESULTS: After treatment, the research group presented significantly lower levels of P, iPTH, and calcium-phosphorus product than the control group, but a significantly higher Ca level than the control group (all P<0.05). In addition, after treatment, the research group showed significantly lower levels of ß2-MG, Scr and BUN but a higher Alb level than the control group (all P<0.05). After treatment, the research group had a greater improvement in immune function-related indexes (IgG and IgM) than the control group (all P<0.05), while the control group had significantly decreased Alb, PA and Hb after treatment (all P<0.05), but the levels of these in the research group did not change greatly (all P>0.05). Risk scoring formula was constructed: risk score = (dialysis time * 0.057123881) + (Ca * -0.100413548) + (P * 0.100419363) + (calcium and phosphorus product * 0.03872268) + (iPTH * 0.000358779). According to inter-group comparison of risk score, the Improvement group got a lower risk score than the Non-improvement group (P<0.0001). Moreover, according to ROC curve-based analysis, the area under the curve of risk score in predicting the efficacy of patients was 0.991. CONCLUSION: Hemodialysis combined with acupuncture and blood perfusion can control the immune regulation by increasing the blood calcium content without affecting nutritional status, but it has no significant effect on the efficacy in patients.

Ethanol extract of propolis regulates type 2 diabetes in mice via metabolism and gut microbiota.

ETHNOPHARMACOLOGICAL RELEVANCE: Propolis is a traditional natural medicine with various activities such as antioxidant and anti-inflammatory, immunomodulatory, anti-tumour, gastroenteritis treatment and prevention, anti-microbial and parasitic, as well as glucose regulation and anti-diabetes, and is expected to be an anti-diabetic candidate with few side effects, but the mechanism of action of propolis on type 2 diabetes mellitus (T2DM) has not been fully elucidated. AIM OF THE STUDY: The purpose of this study was to investigate the mechanism of the effect of ethanol extract of propolis (EEP) on the regulation of blood glucose in T2DM mice. MATERIALS AND METHODS: We studied the possible mechanism of EEP on T2DM using an animal model of T2DM induced by a combination of a high-fat diet and intraperitoneal injection of streptozotocin (STZ). The experiment was divided into four groups, namely, the normal group (HC), model group (T2DM), EEP and metformin group (MET). Biochemical indexes and cytokines were measured, and the differences of metabolites in the serum were compared by 1H-NMR. In addition, the diversity of intestinal flora in feces was studied by 16S rDNA amplicon sequencing. RESULTS: The results showed that following treatment with EEP and MET, the weight-loss trend of mice was alleviated, and the fasting blood glucose, insulin secretion level, insulin resistance index, C peptide level and oral glucose tolerance level decreased, whereas the insulin sensitivity index increased, thereby EEP effectively alleviated the occurrence of T2DM and insulin resistance. Compared with the T2DM group, the concentrations of pro-inflammatory cytokines interleukin-1 beta (IL-1ß), interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) decreased significantly in EEP and MET groups, whereas the concentrations of anti-inflammatory cytokine interleukin-10 (IL-10) increased significantly. Metabolomics results revealed that EEP and MET regulate carbohydrate metabolism and restore amino acid and lipid metabolism. Correlation analysis of intestinal flora in mouse feces showed that compared with the HC group, harmful bacteria such as Bilophila, Eubacterium_ventriosum_group, Mucispirillum and Desulfovibrio were found in the T2DM group, whereas the abundance of beneficial bacteria such as Lactobacillus was significantly reduced. Parabacteroides, Akkermansia, Leuconostoc, and Alloprevotella were abundantly present in the EEP group; however, the MET group showed an increase in the genus Parasutterella, which could regulate energy metabolism and insulin sensitivity. CONCLUSIONS: The results showed that EEP and MET reduce fasting blood glucose in T2DM mice, followed by alleviating insulin resistance, improving the inflammatory reaction of mice, regulating the metabolism of mice, and affecting the steady state of gut microbiota. However, the overall therapeutic effect of EEP is better than that of MET.

Evidence for predictions established by phantom sound.

Predictions, the bridge between the internal and external worlds, are established by prior experience and updated by sensory stimuli. Responses to omitted but unexpected stimuli, known as omission responses, can break the one-to-one mapping of stimulus-response and can expose predictions established by the preceding stimulus built up. While research into exogenous predictions (driven by external stimuli) is often reported, that into endogenous predictions (driven by internal percepts) is rarely available in the literature. Here, we report evidence for endogenous predictions established by the Zwicker tone illusion, a phantom pure-tone-like auditory percept following notch noises. We found that MMN, P300, and theta oscillations could be recorded using an omission paradigm in subjects who can perceive Zwicker tone illusions, but could not in those who cannot. The MMN and P300 responses relied on attention, but theta oscillations did not. In-depth analysis shows that an increase in single-trial theta power, including total and induced theta, with the endogenous prediction, is lateralized to the left frontal brain areas. Our study depicts that the brain automatically analyzes internal perception, progressively establishes predictions and yields prediction errors in the left frontal region when a violation occurs.

Genomic characterization of bZIP transcription factors related to andrographolide biosynthesis in Andrographis paniculata.

The basic leucine zipper (bZIP) transcription factor family plays an important role in various biological processes in plants. Andrographis paniculata (Burm.f) Nees, belonging to the family Acanthaceae, has been widely used as an important traditional herb with a wide range of pharmacological activities, such as antivenom, antiretroviral, anticancer and so on. However, there was no comprehensive analysis of bZIP gene family in the Andrographis paniculata been reported. In this study, we identified 62 bZIPs in Andrographis paniculata and grouped them into 12 subfamilies through the phylogenetic tree analysis. The bZIPs in the same groups have similar motif composition, exon-intron structure and domain distribution. In addition, the RNA-seq data gave a reference for selecting candidate bZIPs to make further function verification. Lastly, qRT-PCR analyses revealed seven ApbZIPs (ApbZIP4, ApbZIP19, ApbZIP30, ApbZIP42, ApbZIP50, ApbZIP52, ApbZIP62) were the most highly expressed in leaf and significantly up-regulated with MeJA and ABA treatment which may be involved in biosynthesis regulation of andrographolide. These data pave the way for further revealing the function of the bZIPs in Andrographis paniculata.

Potent Inhibition of Human Cytochrome P450 3A4 by Biflavone Components from Ginkgo Biloba and Selaginella Tamariscina.

CYP3A4-mediated Phase I biotransformation is the rate-limiting step of elimination for many commonly used clinically agents. The modulatory effects of herbal medicines on CYP3A4 activity are one of the risk factors affecting the safe use of drug and herbal medicine. In the present study, the inhibitory effects of nearly hundred kinds of herbal medicines against CYP3A4 were evaluated based on a visual high-throughput screening method. Furthermore, biflavone components including bilobetin (7-demethylginkgetin, DGK), ginkgetin (GK), isoginkgetin (IGK), and amentoflavone (AMF) were identified as the main inhibitory components of Ginkgo biloba L. (GB) and Selaginella tamariscina (P. Beauv.) Spring (ST), which displayed very strong inhibitory effects toward CYP3A4. The inhibitory effects of these biflavones on clinical drugs that mainly undergo CYP3A4-dependent metabolism were evaluated. The IC 50 of GK toward tamoxifen, gefitinib and ticagrelor were found to be of 0.478 ± 0.003, 0.869 ± 0.001, and 1.61 ± 0.039 µM, respectively. These results suggest the potential pharmacokinetic interactions between the identified biflavones and clinical drugs undergoing CYP3A4-mediated biotransformation. The obtained information is important for guiding the rational use of herbal medicine in combination with synthetic pharmaceuticals.

Functional Characterization of a Novel Glycosyltransferase (UGT73CD1) from Iris tectorum Maxim. for the Substrate promiscuity.

Glycosylflavonoids are a class of natural products with multiple pharmacological activities and a lot of glycosyltransferases from various plant species have been reported that they were involved in the biosynthesis of these phytochemicals. However, no corresponding glycosyltransferase has been identified from the famous horticultural and medicinal plant Iris tectorum Maxim. Here, UGT73CD1, a novel glycosyltransferase, was identified from I. tectorum. based on transcriptome analysis and functional identification. Phylogenetic analysis revealed that UGT73CD1 grouped into the clade of flavonoid 7-OH OGTs. Biochemical analysis showed that UGT73CD1 was able to glycosylate tectorigenin at 7-OH to produce tectoridin, and thus assigned as a 7-O-glycosyltransferase. In addition, it also possessed robust catalytic promiscuity toward 12 structurally diverse flavonoid scaffolds and 3, 4-dichloroaniline, resulting in forming O- and N-glycosides. This work will provide insights into efficient biosynthesis of structurally diverse flavonoid glycosides for drug discovery.

Deficient sensory and cognitive processing in children with cochlear implants: An event-related potential study.

Compared with children having normal hearing (NH), those with cochlear implants (CIs) perform poorly in spoken language comprehension which involves both low-level acoustic encoding and higher-level cognitive processing. Here, we performed an electroencephalography study to portray this brain dynamics of speech perception in CI children. We presented a Mandarin Chinese monosyllable or four-syllable idiom to CI and NH children, and infrequently varied its lexical tone to form a novel monosyllable or pseudo-idiom in an oddball paradigm. The tone contrast embedded in the monosyllables evoked similar mismatch negativities (MMNs) in CI and NH children at an early stage (~200 ms). However, the amplitude of the MMN evoked by the tone contrast in the idiom context was significantly lower in CI children than in NH children. Furthermore, robust late discriminative negativity (LDN) at a late stage (~500 ms) was found only in NH children, but not in CI children. The MMN and LDN findings indicate deficits of low-level acoustic encoding in a complex context (such as an idiom) and higher-level cognitive processing in CI children, respectively. Both deficient sensory and cognitive processing may contribute to the speech perception difficulties in CI children.

Effects of pine pollen wall on gut microbiota and biomarkers in mice with dyslipidemia.

Pinus yunnanensis pollen is rich in various physiological functions. However, whether the pine pollen wall (PW) plays a beneficial role in the body has not been studied. In this work, we have analyzed its effects on the metabolism and gut microbiota of mouse models of dyslipidemia. We found that the intake of pine PW prevents the liver pathologic changes and reduce the concentrations of TNF-α, IL-6, TC, and high-density lipoprotein cholesterol. Moreover, it can regulate bile acid and fat metabolism, SCFAs content, and the structure of the gut microbiota. According to the change of carbohydrate metabolites, we speculated that cellulose should be the main component to play the above beneficial role, and sporopollenin cannot be utilized in the intestine. Therefore, we consider this study of great significance as it gives a description of biological effects of the pine PW and paves the road to its use in health products.

Efficacy of Joungal in preventing febrile neutropenia induced by platinum-based doublet chemotherapy in lung cancer.

BACKGROUND: Myelosuppressive chemotherapy often results in febrile neutropenia (FN) in patients with lung cancer, resulting in infection, prolonged hospitalization, higher economic and labor costs, and increased mortality rate. Colony-stimulating factor (CSF) is used to treat FN, but it exhibits limited efficacy and is often underused. We evaluated Joungal, a traditional Chinese medicine, for treatment of neutropenic complications in patients with lung cancer who received chemotherapy. METHODS: A total of 795 patients with lung cancer were treated with platinum-based chemotherapy from 2012 to 2017. Of these, 191 received Joungal during chemotherapy. Three hundred eighty-two patients were included in the control group. The primary end point was incidence of FN. The secondary end points were incidence of neutropenia, granulocyte colony-stimulating factor (G-CSF) use, hospitalization duration, and cost. RESULTS: There were no differences in clinicopathological characteristics such as gender, age, smoking status, stage of disease, hemoglobin, or histologic type between two groups. Joungal significantly decreased the incidence of chemotherapy-induced FN (2.1% vs. 9.4%, OR =0.21, P=0.002), grade 2/3/4 neutropenia (29.8 % vs. 55.8%, OR =0.34, P=0.000), and grade 3/4 neutropenia (13.1% vs. 23.8%, OR =0.48, P=0.013) compared with controls. Furthermore, Joungal decreased G-CSF use (0.68 vs. 1.34/patient/cycle, P=0.001), hospitalization duration (2.56 vs. 4.68 day/patient/cycle, P=0.002), and economic burden ($660 vs. $1,580/ patient/cycle, P=0.001). No drug-related toxicity was observed. CONCLUSIONS: Joungal safely and effectively decreased the incidence of neutropenia and FN induced by doublet platinum-based chemotherapy in patients with lung cancer, and may have potential as a supportive care agent for patients with lung cancer.

Protocol development for discovery of angiogenesis inhibitors via automated methods using zebrafish.

Their optical clarity as larvae and embryos, small size, and high fecundity make zebrafish ideal for whole animal high throughput screening. A high-throughput drug discovery platform (HTP) has been built to perform fully automated screens of compound libraries with zebrafish embryos. A Tg(kdrl:EGFP) line, marking endothelial cell cytoplasm, was used in this work to help develop protocols and functional algorithms for the system, with the intent of screening for angiogenesis inhibitors. Indirubin 3' Monoxime (I3M), a known angiogenesis inhibitor, was used at various concentrations to validate the protocols. Consistent with previous studies, a dose dependant inhibitory effect of I3M on angiogenesis was confirmed. The methods and protocols developed here could significantly increase the throughput of drug screens, while limiting human errors. These methods are expected to facilitate the discovery of novel anti-angiogenesis compounds and can be adapted for many other applications in which samples have a good fluorescent signal.

Development of Industrial Oil Crop Crambe abyssinica for Wax Ester Production through Metabolic Engineering and Cross Breeding.

As an important industrial feedstock, wax esters (WEs) have been used as lubricants in a number of technical processes. There is however currently no large-scale biological source for WE production and alteration in metabolic pathways of plant oils for producing WEs could be attractive to the commercial markets. Here, we present the breeding results of long-term studies on successful development of new crambe lines producing WEs through genetic engineering and cross breeding. The transgenic crambe lines producing WEs at over 25% of the total seed oil were first generated by introduction of the jojoba WE biosynthetic genes ScFAR and ScWS. Further improvement of the lines aiming at improving oxidative stability of WEs was achieved through introducing the CaFAD2-RNAi gene into these lines by crossing. The hybrid lines possessed similar agronomic traits to the wild type and a stable level of WEs over several generations, suggesting a high potential of crambe as an industrial crop for WE production.

Acupuncture through Baihui (DU20) to Qubin (GB7) mitigates neurological impairment after intracerebral hemorrhage.

Inflammation plays an important role in nerve defects caused by intracerebral hemorrhage. Repairing brain damage by inhibiting the macrophage-inducible C-type lectin/spleen tyrosine kinase (Mincle/Syk) signaling pathway is a potential new target for treating cerebral hemorrhage. In this study, we aimed to determine whether acupuncture through Baihui (DU20) to Qubin (GB7) is an effective treatment for intracerebral hemorrhage through the Mincle/Syk signaling pathway. An intracerebral hemorrhage rat model was established by autologous blood infusion into the caudate nucleus. Acupuncture through Baihui to Qubin was performed for 30 minutes, once every 12 hours, for a total of three times. Piceatannol (34.62 mg/kg), a Syk inhibitor, was intraperitoneally injected as a control. Modified neurological severity score was used to assess neurological function. Brain water content was measured. Immunohistochemistry and western blot assay were used to detect immunoreactivity and protein expression levels of Mincle, Syk, and CARD9. Real-time polymerase chain reaction was used to determine interleukin-1ß mRNA levels. Hematoxylin-eosin staining was performed to observe histopathological changes. Our results showed that acupuncture through Baihui to Qubin remarkably improved neurological function and brain water content, and inhibited immunoreactivity and expression of Mincle, Syk, CARD9, and interkeukin-1ß. Moreover, this effect was similar to piceatannol. These findings suggest that acupuncture through Baihui to Qubin can improve neurological impairment after cerebral hemorrhage by inhibiting the Mincle/Syk signaling pathway.

Draft genome sequence of the Tibetan medicinal herb Rhodiola crenulata.

Rhodiola crenulata, a well-known medicinal Tibetan herb, is mainly grown in high-altitude regions of the Tibet, Yunnan, and Sichuan provinces in China. In the past few years, increasing numbers of studies have been published on the potential pharmacological activities of R. crenulata, strengthening our understanding into its putitive active ingredient composition, pharmacological activity, and mechanism of action. These findings also provide strong evidence supporting the important medicinal and economical value of R. crenulata. Consequently, some Rhodiola species are becoming endangered because of overexploitation and environmental destruction. However, little is known about the genetic and genomic information of any Rhodiola species. Here we report the first draft assembly ofthe R. crenulata genome, which was 344.5 Mb (25.7 Mb Ns), accounting for 82% of the estimated genome size, with a scaffold N50 length of 144.7 kb and a contig N50 length of 25.4 kb. The R. crenulata genome is not only highly heterozygous but also highly repetitive, with ratios of 1.12% and 66.15%, respectively, based on the k-mer analysis. Furthermore, 226.6 Mb of transposable elements were detected, of which 77.03% were long terminal repeats. In total, 31 517 protein-coding genes were identified, capturing 86.72% of expected plant genes in BUSCO. Additionally, 79.73% of protein-coding genes were functionally annotated. R. crenulata is an important medicinal plant and also a potentially interesting model species for studying the adaptability of Rhodiola species to extreme environments. The genomic sequences of R. crenulata will be useful for understanding the evolutionary mechanism of the stress resistance gene and the biosynthesis pathways of the different medicinal ingredients, for example, salidroside in R. crenulata.

Responses of Organic Phosphorus Fractionation to Environmental Conditions and Lake Evolution.

Geochemical fractionation is used to assess the significance of environmental factors on organic phosphorus (OP) pools in sediments. Labile, moderately labile, and nonlabile OP pools in the sediments from Lake Hulun, Inner Mongolia, were fractionated, and their responses to environmental conditions and lake evolution were investigated based on the spatial and vertical distribution of OP fractionations. In light of the recalcitrant characteristics of organic matter (OM) in different environmental conditions, the pH presents significant negative effects on the amount of labile OP, while water depth shows an important role in regulating the distribution between the moderately labile and nonlabile OP pools. A latitudinal zonation in the distribution of OP pools in surface sediments from different lakes was apparent with this zonation likely linked to the gradient effects of climate and anthropogenic activities on OM decomposition and thereby on the sediments capacity to hold phosphorus. These results show that OM plays a role in governing the impacts of weather and environmental factors on OP fractionation in aquatic environments. This work suggests that OP pools in the sediment core could be used as an archive for environmental conditions and lake evolution.

[Cloning and expression analysis of a chitinase gene PnCHI1 from Panax notoginseng].

Chitinases(EC3.2.1.14), which are present in various organisms, catalyze the hydrolytic cleavage of chitin and play a vital role in plant defense mechanisms against fungal pathogens.In addition, the chitinases are well known to regulate plant growth and development and are involved in programmed cell death(PCD).A chitinase expressed sequence tag(EST) was isolated from Panax notoginseng, and the full-length cDNA of this EST was cloned with the method of rapid amplification of cDNA ends and named as PnCHI1. PnCHI1 was 1 022 bp in length and contained an intact open reading frame(ORF) of 822 bp, a 26 bp 5'-untranslated region(UTR), and a 174 bp 3'-UTR.The predicted protein of PnCHI1 with 273 amino acid residues belongs to glycoside hydrolase family 19 and fell into the class IV of chitinases through phylogenetic analysis.QRT-PCR analysis showed that the expression of PnCHI1 was induced by methyl jasmonate, ethylene, H2O2, and salicylic acid.PnCHI1 was quickly induced after inoculation with Alternaria panax.Moreover, the expression level of PnCHI1 was increased after pretreatment with methyl jasmonate, and then the transcription level of PnCHI1was sharp increased after inoculation with Fusarium solani,and the highest transcription level was achieved at 4 h post inoculation.But the expression level of PnCHI1 in the sterile water pretreated P.notoginseng was increased gradually after inoculation with F.solani, and the highest expression level was achieved at 48 h post inoculation.All the results of present study indicated that PnCHI1 was involved in defense response of P.notoginseng against the F.solani and A.panax.

Distribution characteristics of organic phosphorus in sediments from Lake Hulun, China.

The amount of organic phosphorus (OP) and its distribution among different pools in lake sediments depend on biotic and abiotic processes driving OP fractionation. The key abiotic factors governing these transformation processes between different OP fractions in sediments were studied on the basis of distribution characteristics of OP pools in sediments from Lake Hulun (HLH). The results showed the order of the size of OP pools in the surface sediments from Lake Hulun to be: Re-OP (residual OP) ≫ FAOP (fulvic acid fraction) ≥ HCl-OP (OP extracted by HCl) > HAOP (humic acid fraction) ≫ LOP (labile OP); lower concentrations of LOP were found in lake surface sediments with high pH (pH > 9) of lake overlying water indicating a governing role of pH in LOP degradation in an aquatic environment. The pattern of total OP (TOP) spatial distribution showed an obvious decreasing trend from the west to the east lake districts in surface sediments from HLH, which was attributed to the inputs of dust and dry grass driven by the prevailing wind and the finer grain size in the west lake districts. However, the spatial distribution pattern of dissolved OP (DOP) in overlying water, which presented different trends with TOP and total organic carbon (TOC) in surface sediments, indicated that the biological factors and terrestrial inputs showed a joint influence on DOP distribution in HLH. The higher levels of Re-OP and lower levels of HCl-OP observed in HLH may reveal that calcium ions and their minerals were the key governing factors of OP fractionation in sediments from HLH. This work revealed the main abiotic process or mechanism driving OP fractionation in the aquatic environment helping to understand the geochemical information archived in OP pools in lake sediments.

RNAi targeting putative genes in phosphatidylcholine turnover results in significant change in fatty acid composition in Crambe abyssinica seed oil.

The aim of this study was to evaluate the importance of three enzymes, LPCAT, PDCT and PDAT, involved in acyl turnover in phosphatidylcholine in order to explore the possibility of further increasing erucic acid (22:1) content in Crambe seed oil. The complete coding sequences of LPCAT1-1 and LPCAT1-2 encoding lysophosphatidylcholine acyltransferase (LPCAT), PDCT1 and PDCT2 encoding phosphatidylcholine:diacylglycerol cholinephosphotransferase (PDCT), and PDAT encoding phospholipid:diacylglycerol acyltransferase (PDAT) were cloned from developing Crambe seeds. The alignment of deduced amino acid sequences displayed a high similarity to the Arabidopsis homologs. Transgenic lines expressing RNA interference (RNAi) targeting either single or double genes showed significant changes in the fatty acid composition of seed oil. An increase in oleic acid (18:1) was observed, to varying degrees, in all of the transgenic lines, and a cumulative effect of increased 18:1 was shown in the LPCAT-PDCT double-gene RNAi. However, LPCAT single-gene RNAi led to a decrease in 22:1 accumulation, while PDCT or PDAT single-gene RNAi had no obvious effect on the level of 22:1. In agreement with the abovementioned oil phenotypes, the transcript levels of the target genes in these transgenic lines were generally reduced compared to wild-type levels. In this paper, we discuss the potential to further increase the 22:1 content in Crambe seed oil through downregulation of these genes in combination with fatty acid elongase and desaturases.

Bottlenecks in erucic acid accumulation in genetically engineered ultrahigh erucic acid Crambe abyssinica.

Erucic acid is a valuable industrial fatty acid with many applications. The main producers of this acid are today high erucic rapeseed (Brassica napus) and mustard (Brassica juncea), which have 45%-50% of erucic acid in their seed oils. Crambe abyssinica is an alternative promising producer of this acid as it has 55%-60% of erucic acid in its oil. Through genetic modification (GM) of three genes, we have previously increased the level of erucic acid to 71% (68 mol%) in Crambe seed oil. In this study, we further investigated different aspects of oil biosynthesis in the developing GM Crambe seeds in comparison with wild-type (Wt) Crambe, rapeseed and safflower (Carthamus tinctorius). We show that Crambe seeds have very low phosphatidylcholine-diacylglycerol interconversion, suggesting it to be the main reason why erucic acid is limited in the membrane lipids during oil biosynthesis. We further show that GM Crambe seeds have slower seed development than Wt, accompanied by slower oil accumulation during the first 20 days after flowering (DAF). Despite low accumulation of erucic acid during early stages of GM seed development, nearly 86 mol% of all fatty acids accumulated between 27 and 50 DAF was erucic acid, when 40% of the total oil is laid down. Likely bottlenecks in the accumulation of erucic acid during early stages of GM Crambe seed development are discussed.

Development of ultra-high erucic acid oil in the industrial oil crop Crambe abyssinica.

Erucic acid (22 : 1) is a major feedstock for the oleochemical industry. In this study, a gene stacking strategy was employed to develop transgenic Crambe abyssinica lines with increased 22 : 1 levels. Through integration of the LdLPAAT, BnFAE1 and CaFAD2-RNAi genes into the crambe genome, confirmed by Southern blot and qRT-PCR, the average levels of 18 : 1, 18 : 2 and 18 : 3 were markedly decreased and that of 22 : 1 was increased from 60% in the wild type to 73% in the best transgenic line of T4 generation. In single seeds of the same line, the 22 : 1 level could reach 76.9%, an increase of 28.0% over the wild type. The trierucin amount was positively correlated to 22 : 1 in the transgenic lines. Unlike high erucic rapeseed, the wild-type crambe contains 22 : 1 in the seed phosphatidylcholine and in the sn-2 position of triacylglycerols (5% and 8%, respectively). The transgenic line with high 22 : 1 had decreased 22 : 1 level in phosphatidylcholine, and this was negatively correlated with the 22 : 1 level at the sn-2 position of TAG. The significances of this study include (i) achieving an unprecedented level of 22 : 1 in an oil crop; (ii) disclosing mechanisms in the channelling of a triacylglycerol-specific unusual fatty acid in oil seeds; (iii) indicating potential limiting factors involved in the erucic acid biosynthesis and paving the way for further increase of this acid and (iv) development of an added value genetically modified oil crop having no risk of gene flow into feed and food crops.

[Effects of rosuvastatin on monocrotaline-induced pulmonary artery hypertension in rats].

OBJECTIVE: To investigate the effects of rosuvastatin on monocrotaline (MCT)-induced pulmonary artery hypertension in rats. METHODS: Pulmonary arterial hypertension was induced by a single subcutaneous injection of monocrotaline (50 mg/kg) in rats. In the prevention protocol, 32 male Sprague-Dawley rats were randomly divided into four groups (n = 8 each): low-dose rosuvastatin prevention group (2 mg×kg(-1)×d(-1)), high-dose rosuvastatin prevention group (10 mg×kg(-1)×d(-1)), pulmonary arterial hypertension group, normal control group. Beginning on the MCT injection day, rats were treated with rosuvastatin by daily gavage for 4 weeks. Normal control group and pulmonary arterial hypertension group received vehicle by gavage. In the treatment protocol, 52 male Sprague-Dawley rats were randomly divided into four groups (n = 13 each): low-dose rosuvastatin treatment group (2 mg×kg(-1)×d(-1)), high-dose rosuvastatin treatment group (10 mg×kg(-1)×d(-1)), pulmonary arterial hypertension group, normal control group. Four weeks after MCT injection, rats were treated with rosuvastatin by daily gavage for 4 weeks. Normal control group and pulmonary arterial hypertension group received vehicle by gavage. At the end of study, survival rates, mean pulmonary arterial pressure (mPAP), wall thickness of small pulmonary artery and right ventricular hypertrophy among groups were compared. The expression levels of proliferating cell nuclear antigen (PCNA) and endothelial nitricoxide synthase (eNOS) protein in small pulmonary artery, the expression levels of Rho kinase 1(ROCK-1) and eNOS mRNA in lung tissue were also detected. RESULTS: All rats in the prevention protocol survived. Rosuvastatin treatment improved survival in the treatment protocol (58%, 75% vs.30%, P < 0.05). Rosuvastatin therapy in both preventive or treatment protocols significantly lowered mPAP [prevention protocol: (27.53 ± 3.43), (25.72 ± 1.76) vs. (36.05 ± 2.45) mm Hg (1 mm Hg = 0.133 kPa), P < 0.01; treatment protocol: (30.39 ± 3.17), (27.59 ± 1.99) vs. (40.68 ± 1.39) mm Hg, P < 0.01], reduced thickening of small pulmonary artery wall (P < 0.01) and right ventricular hypertrophy (P < 0.01). Rosuvastatin also inhibited PCNA expression of SMC (P < 0.01), restored eNOS expression of EC (P < 0.05) and inhibited ROCK-1 mRNA expressions in lung tissue (P < 0.05). CONCLUSIONS: Rosuvastatin therapy reduced mPAP in monocrotaline-induced pulmonary arterial hypertension rat model and this effect is linked with inhibition of ROCK-1 expression, inhibition of smooth muscle cell proliferation and restoration of endothelial cell functions.