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The pharmacological mechanism of curcumin against drug resistance in non-small cell lung cancer (NSCLC) remains unclear. This study aims to summarize the genes and pathways associated with curcumin action as an adjuvant therapy in NSCLC using network pharmacology, drug-likeness, pharmacokinetics, functional enrichment, protein-protein interaction (PPI) analysis, and molecular docking. Prognostic genes were identified from the curcumin-NSCLC intersection gene set for the following drug sensitivity analysis. Immunotherapy, chemotherapy, and targeted therapy sensitivity analyses were performed using external cohorts (GSE126044 and IMvigor210) and the CellMiner database. 94 curcumin-lung adenocarcinoma (LUAD) hub targets and 41 curcumin-lung squamous cell carcinoma (LUSC) hub targets were identified as prognostic genes. The anticancer effect of curcumin was observed in KEGG pathways involved with lung cancer, cancer therapy, and other cancers. Among the prognostic curcumin-NSCLC intersection genes, 20 LUAD and 8 LUSC genes were correlated with immunotherapy sensitivity in the GSE126044 NSCLC cohort; 30 LUAD and 13 LUSC genes were associated with immunotherapy sensitivity in the IMvigor210 cohort; and 12 LUAD and 13 LUSC genes were related to chemosensitivity in the CellMiner database. Moreover, 3 LUAD and 5 LUSC genes were involved in the response to targeted therapy in the CellMiner database. Curcumin regulates drug sensitivity in NSCLC by interacting with cell cycle, NF-kappa B, MAPK, Th17 cell differentiation signaling pathways, etc. Curcumin in combination with immunotherapy, chemotherapy, or targeted drugs has the potential to be effective for drug-resistant NSCLC. The findings of our study reveal the relevant key signaling pathways and targets of curcumin as an adjuvant therapy in the treatment of NSCLC, thus providing pharmacological evidence for further experimental research.

Cancer progression and metastases are the leading causes of poor outcomes in patients with colon cancer. Colon cancer metastasis is a multigene, multistep, multistage complex process in which target genes, microRNAs, epithelial-stromal transformation, tumour stem cells, the tumour microenvironment, and various cell signalling pathways are implicated in the progression and metastasis of colon cancer. Although conventional therapies have made significant advances in treating the progression and metastasis of colorectal cancer, they have failed to improve survival outcomes. Natural compounds may have more significant potential in preventing and treating colon cancer. Active natural compounds exert their antitumor effects by inducing tumour cell differentiation, promoting tumour cell apoptosis, inhibiting tumour vascular growth, and regulating immunity. Natural compounds, combined with conventional therapies, can target mutant genes and various cellular signalling pathways, inhibit epithelial-stromal transformation, and improve the tumour microenvironment to inhibit tumour progression and metastasis. The synergism of natural compounds and conventional therapeutics has the potential to become a promising therapy for treating colorectal cancer progression and metastases.

Background. Compound Taxus chinensis capsule (CTCC), an antitumor Chinese patent medicine, has been commonly prescribed as an adjunctive agent to chemotherapy for the management of non-small-cell lung cancer (NSCLC); however, the effects of CTCC added to chemotherapy for NSCLC patients have never been comprehensively evaluated or summarized. Purpose. To assess the synergistic effects of CTCC and chemotherapy on NSCLC. Study Design. Evidence-based study, systematic review, and quantitative meta-analysis. Methods. This systematic review and meta-analysis was implemented in accordance with the PRISMA (Preferred Reported Items for Systematic Review and Meta-Analysis) guidelines. Eight databases including China National Knowledge Infrastructure, SINOMED, China Biomedical Literature Database, Wanfang Database, VIP, PubMed, Cochrane Library, and EMBASE were searched for relevant RCTs from their inception until May 24, 2021, and hand-searching was also carried out to identify additional studies. All randomized controlled trials (RCTs) that compared CTCC combined with chemotherapy versus chemotherapy alone were included in our study. The Cochrane Risk-of-Bias tool was used to determine the risk of bias and methodological quality of the included RCTs. Review Manager 5.3 software was used for comprehensive analysis. The primary outcome measure for this study was the disease control rate (DCR), and the secondary outcomes included the objective response rate (ORR), adverse reactions, and quality of life (QOL). Results. Six RCTs with a total sample size of 410 were finally included. The pooled data showed that, compared with chemotherapy alone, CTCC combined with chemotherapy significantly improved DCR (RR = 1.15, 95% CI: 1.06–1.25, P = 0.006), ORR (RR = 1.38, 95% CI: 1.18–1.63, P P P P = 0.004), leukopenia ( P = 0.0009), thrombocytopenia ( P = 0.01), rash ( P = 0.002), and fever ( P = 0.007). Conclusion. Based on the available evidence, compared with chemotherapy alone, CTCC used as an adjunctive agent to chemotherapy for NSCLC can improve the clinical efficacy and quality of life and decrease the likelihood of adverse reactions, suggesting that CTCC might be an effective and safe adjunctive medicine to chemotherapy for NSCLC. However, considering the relatively small sample size and the inherent imperfections of the included randomized controlled trials, more high-quality clinical trials with longer follow-up time are needed to further assess the efficacy and safety of this combined treatment regimen.

Some lipid digestion pathways in fish deviate from those in mammals, and many differences may also be species dependent. This report describes a pathway for monoacylglycerol (MAG) and lysophospholipid absorption by intestinal enterocytes in brown trout that may be of significance in salmonids. When culturing primary cells in a medium containing 1- and 2-MAG, we observed a massive hydrolysis of unesterified fatty acids. The hydrolysis activity was retained in the medium even after the removal of the cells. To further characterize these activities, both extracellular and isolated membrane proteins were tested for lipase activity toward triacylglycerol (TAG), diacylglycerol (DAG), MAG, phosphatidylcholine (PtdCho), and lysoPtdCho. In both cases, the main hydrolyzing activity was toward MAG followed by lysoPtdCho with very little activity toward DAG, TAG, or PtdCho. The extracellular and membrane proteins were partially purified by fast protein liquid chromatography and identified by proteomics (liquid chromatography-tandem mass spectrometry) focusing on lipase/hydrolase enzymes. In the membrane protein fraction, the data suggested that MAG was produced as an intermediate in the hydrolysis of lysoPtdCho by either lysophospholipase C or lysophospholipase D activity. Both abhydrolase-domain-containing protein 6 and abhydrolase-domain-containing protein 12 were identified in the membrane protein and they could be responsible for the hydrolysis of MAG. In the culture medium, low-peptide matches were found for ABHD6 and phospholipases and further studies are needed. In summary, trout enterocytes are capable of hydrolyzing MAG and lysoPtdCho. The enzymes are both extracellular and membrane bound. The pathways may be of significance during lipid absorption in fish lacking a 1,3 specific pancreatic lipase.

Domestication of Salmo salar has imposed strong selection for production traits since the 1970s. The domestication has also imposed a radical shift in diet. Whereas wild salmon eats invertebrates, crustaceans and fish, the dietary lipids in domestic feed has since 1990 gradually shifted from fish oil (FO) to vegetable oil (VO), causing a decrease intake of long-chain polyunsaturated fatty acids (LC-PUFA). We tested the hypothesis that this shift has induced domestication-specific features of lipid metabolism in a 96-day feeding trial of domesticated and wild salmon fed diets based on FO, VO or phospholipids (PL). We addressed this by sampling tissues central in fat uptake (pyloric caeca) and processing (liver) and quantifying RNA expression and fatty acid composition. Domesticated salmon grew faster than wild salmon, with higher gene expression in glucose and lipid metabolism pathways. The promoters of differentially expressed genes were enriched for transcription factors involved in circadian clock regulation. Domesticated salmon had lower expression of cry2 and nr1d1, genes involved in negative regulation of circadian rhythm, with possible implications for the diurnal cycle of feed ingestion and basal metabolic rate. Only wild salmon showed a significant impact on growth of VO versus PL or FO feed, whereas domesticated but not wild salmon upregulated key LC-PUFA synthesis genes fads2d5 and fads2d6a in response to VO diet. Domesticated salmon had higher LC-PUFA but lower 18:3n-3 and 18:2n-6 in liver when fed VO, suggesting that domesticated salmon can better compensate for dietary shortage of LC-PUFA compared to wild salmon.

Atlantic salmon can synthesize polyunsaturated fatty acids (PUFAs), such as eicosapentaenoic acid (20:5n-3), arachidonic acid (20:4n-6) and docosahexaenoic acid (22:6n-3) via activities of very long chain fatty acyl elongases (Elovls) and fatty acyl desaturases (Fads), albeit to a limited degree. Understanding molecular mechanisms of PUFA biosynthesis and regulation is a pre-requisite for sustainable use of vegetable oils in aquafeeds as current sources of fish oils are unable to meet increasing demands for omega-3 PUFAs. By generating CRISPR-mediated elovl2 partial knockout (KO), we have shown that elovl2 is crucial for multi-tissue synthesis of 22:6n-3 in vivo and that endogenously synthesized PUFAs are important for transcriptional regulation of lipogenic genes in Atlantic salmon. The elovl2-KOs showed reduced levels of 22:6n-3 and accumulation of 20:5n-3 and docosapentaenoic acid (22:5n-3) in the liver, brain and white muscle, suggesting inhibition of elongation. Additionally, elovl2-KO salmon showed accumulation of 20:4n-6 in brain and white muscle. The impaired synthesis of 22:6n-3 induced hepatic expression of sterol regulatory element binding protein-1 (srebp-1), fatty acid synthase-b, Δ6fad-a, Δ5fad and elovl5. Our study demonstrates key roles of elovl2 at two penultimate steps of PUFA synthesis in vivo and suggests Srebp-1 as a main regulator of endogenous PUFA synthesis in Atlantic salmon.

sn-2-Monoacylglycerol (2-MAG) and sn-1(3)-monoacylglycerol [1(3)-MAG] are important but yet little studied intermediates in lipid metabolism. The current study compared the metabolic fate of 2-MAG and 1(3)-MAG in isolated caecal enterocytes and hepatocytes of brown trout (Salmo trutta). 1(3)-Oleoyl [9,10-3H(N)]-glycerol and 2-Oleoyl [9,10-3H(N)]-glycerol were prepared by pancreatic lipase digestion of triolein [9,10-3H(N)]. The 1(3)-MAG and 2-MAG were efficiently absorbed by enterocytes and hepatocytes at similar rates. The 2-MAG was quickly resynthesized into TAG through the monoacylglycerol acyltransferase (EC: 2.3.1.22, MGAT) pathway in both tissues, whereas 1(3)-MAG was processed into TAG and phospholipids at a much slower rate, suggesting 2-MAG was the preferred substrates for MGAT. Further analysis showed that 1(3)-MAG was synthesized into 1,3-DAG, but there were no accumulation of 1,3-DAG in either enterocytes or hepatocytes, which contrasts that of mammalian studies. Some of the 1(3)-MAG may be acylated to 1,2(2,3)-DAG and then utilized for TAG synthesis. Alternatively, 1(3)-MAG can be hydrolyzed to free fatty acid and glycerol, and re-synthesized into TAG through the glycerol-3-phosphate (Gro-3-P) pathway. The overall data suggested that the limiting step of the intracellular 1(3)-MAG metabolism is the conversion of 1(3)-MAG itself.

The mechanism of essentiality of dietary phospholipid (PL) for larval fish is not clear. The main objective of the present study was to determine if the PL requirement of Atlantic cod larvae was due to any genetic impairment caused by functional immaturity. Cod larvae were sampled at 1, 3, 8, 13, 17, 18, 30, 42 and 60 days post hatch (dph) for transcriptome analysis using a recently developed microarray. The fatty acid profile and gene expression levels of cod larvae at 17 dph were compared after feeding differently enriched rotifers, which contained different DHA levels in PL. No significant differences (p

We have previously shown that K-selection and microbial stability in the rearing water increases survival and growth of Atlantic cod (Gadus morhua) larvae, and that recirculating aquaculture systems (RAS) are compatible with this. Here, we have assessed how water treatment influenced the larval microbiota and host responses at the gene expression level. Cod larvae were reared with two different rearing water systems: a RAS and a flow-through system (FTS). The water microbiota was examined using a 16S rDNA PCR/DGGE strategy. RNA extracted from larvae at 8, 13, and 17 days post hatching was used for microbiota and microarray gene expression analysis. Bacterial cDNA was synthesized and used for 16S rRNA amplicon 454 pyrosequencing of larval microbiota. Both water and larval microbiota differed significantly between the systems, and the larval microbiota appeared to become more dissimilar between systems with time. In total 4 phyla were identified for all larvae: Actinobacteria, Bacteroidetes, Firmicutes, and Proteobacteria. The most profound difference in larval microbiota was a high abundance of Arcobacter (Epsilonproteobacteria) in FTS larvae (34 ± 9% of total reads). Arcobacter includes several species that are known pathogens for humans and animals. Cod larval transcriptome responses were investigated using an oligonucleotide gene expression microarray covering approximately 24,000 genes. Interestingly, FTS larvae transcriptional profiles revealed an overrepresentation of upregulated transcripts associated with responses to pathogens and infections, such as c1ql3-like, pglyrp-2-like and zg16, compared to RAS larvae. In conclusion, distinct water treatment systems induced differences in the larval microbiota. FTS larvae showed up-regulation of transcripts associated with responses to microbial stress. These results are consistent with the hypothesis that RAS promotes K-selection and microbial stability by maintaining a microbial load close to the carrying capacity of the system, and ensuring long retention times for both bacteria and water in the system. Copyright © 2018 Vestrum, Attramadal, Winge, Li, Olsen, Bones, Vadstein and Bakke. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

Facing a bottleneck in the growth of aquaculture, and a gap in the supply and demand of the highly beneficial n-3 long-chain PUFA (LC-PUFA), sustainable alternatives to traditional marine-based feeds are required. Therefore, in the present trial, a novel oil obtained from a genetically engineered oilseed crop, Camelina sativa, that supplied over 25 % n-3 LC-PUFA was tested as a sole dietary-added lipid source in Atlantic salmon (Salmo salar) feed. Three groups of fish were fed three experimental diets for 12 weeks with the same basal composition and containing 20 % added oil supplied by either a blend of fish oil and rapeseed oil (1:3) (COM) reflecting current commercial formulations, wild-type Camelina oil (WCO) or the novel transgenic Camelina oil (TCO). There were no negative effects on the growth, survival rate or health of the fish. The whole fish and flesh n-3 LC-PUFA levels were highest in fish fed TCO, with levels more than 2-fold higher compared with those of fish fed the COM and WCO diets, respectively. Diet TCO had no negative impacts on the evaluated immune and physiological parameters of head kidney monocytes. The transcriptomic responses of liver and mid-intestine showed only mild effects on metabolism genes. Overall, the results clearly indicated that the oil from transgenic Camelina was highly efficient in supplying n-3 LC-PUFA providing levels double that obtained with a current commercial standard, and similar to those a decade ago before substantial dietary fishmeal and oil replacement. This article has been accepted for publication in British Journal of Nutrition . This version is free to view and download for private research and study only. Not for re-distribution, re-sale or use in derivative works. © The Authors 2018

Recent studies have shown that fish larvae require not only a certain quantitative amount of dietary phospholipid (PL) in their feed, but they also depend on the quality of the dietary PL, and their docosahexaenoic acid (DHA) content is particularly important for normal growth and functional development. The most commonly used live feeds in aquaculture, rotifers (Brachionus sp.) and brine shrimp (Artemia sp.), do not contain adequate amounts of DHA in their PL. Therefore, there is an emerging need to learn more on how PL of live feed organisms can be efficiently enriched. In this chapter, we discussed the factors that could affect the enrichment of DHA in PL of live feed and suggested some strategies that could increase the DHA levels in PL of rotifers. The mechanism behind the PL requirement of fish larvae is not well understood and the overall objective of our studies has been to obtain more knowledge of ontogenesis of PL synthesis capability of early stages of Atlantic cod Gadus morhua. Transcriptome analysis of larvae in different stages was carried out using microarray, to evaluate the effect of development on the expression of key genes of PL biosynthesis. Moreover, labeled lipid precursors were tube fed to cod larvae to evaluate their capacity of PL synthesis. The larvae showed relatively high biosynthesis ability of PL compared to neutral lipids. Our overall data suggested that besides the possible limited de novo PL synthesis ability in the intestine, other metabolic constraints should also be considered.

An increasing number of studies suggest that DHA associated with dietary phospholipid (PL) is better metabolically available for fish larvae. Knowledge on how to increase DHA levels in the major PL classes of the live feed is needed. Brachionus Cayman was cultured in conical fibreglass vessels, enriched with commonly used commercial diet Multigain or DHA Selco (DSelco) in flat bottom vessels at 20 °C. The changes of DHA content in the PL of the rotifers through enrichment and through a starvation phase post-enrichment were investigated in order to define an appropriate enrichment time and to test the stability of DHA post-enrichment. The different dietary non-HUFA (non-highly unsaturated fatty acid) composition of DSelco (25.1% SFA, 32.1% MUFA) and Multigain (40.7% SFA, 2.11% MUFA) allowed an evaluation on how SFA and MUFA might affect the efficiency of DHA enrichment of phospholipid in the rotifers. Rotifer samples were collected at 0 h, 1 h, 2 h, 4 h, 8 h, 12 h and 24 h after initiation of enrichment. Total lipid (TL) and the fatty acid composition in TL, phosphatidylcholine (PC) and phosphatidylethanolamine (PE) of rotifers and enrichment diets were analysed. The highest DHA levels in PC (23.0%) and PE (16.5%) in Cayman were found after 24 h enrichment with Multigain, and levels were far higher than obtained earlier. The enriched DHA was stable at 10 °C for at least 24 h post-enrichment under starving conditions ( p > 0.05), whereas a significant ( p Statement of relevance The paper is bringing information about fatty acid enrichment of rotifers in questions that are so far quite unclear and could have a strong impact on larviculture of marine fish larvae. This study shows methods to obtain high level of DHA in phospholipids of rotifer. The results could potentially have a significant impact to improve current fatty acid enrichment protocols for rotifers.

The main objective was to study time kinetics of change in important highly unsaturated fatty acids (HUFAs) in phosphatidylcholine (PC) and phosphatidylethanolamine (PE) of Artemia franciscana nauplii and juveniles following enrichment and subsequent starvation. Samples of Artemia nauplii were taken at variable times (0.5–24 h) following enrichment and starvation. Samples of Artemia juveniles were taken after 2, 3 and 4 days of cultivation. No docosahexaenoic acid (DHA) was found in PC and PE of Artemia nauplii during the first hour of enrichment, while a significant (P

New de novo sources of omega 3 (n-3) long chain polyunsaturated fatty acids (LC-PUFA) are required as alternatives to fish oil in aquafeeds in order to maintain adequate levels of the beneficial fatty acids, eicosapentaenoic and docosahexaenoic (EPA and DHA, respectively). The present study investigated the use of an EPA+DHA oil derived from transgenic Camelina sativa in Atlantic salmon (Salmo salar) feeds containing low levels of fishmeal (35%) and fish oil (10%), reflecting current commercial formulations, to determine the impacts on tissue fatty acid profile, intestinal transcriptome, and health of farmed salmon. Post-smolt Atlantic salmon were fed for 12-weeks with one of three experimental diets containing either a blend of fish oil/rapeseed oil (FO), wild-type camelina oil (WCO) or transgenic camelina oil (DCO) as added lipid source. The DCO diet did not affect any of the fish performance or health parameters studied. Analyses of the mid and hindgut transcriptomes showed only mild effects on metabolism. Flesh of fish fed the DCO diet accumulated almost double the amount of n-3 LC-PUFA than fish fed the FO or WCO diets, indicating that these oils from transgenic oilseeds offer the opportunity to increase the n-3 LC-PUFA in farmed fish to levels comparable to those found a decade ago. © 2017 Betancor et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Rotifer cultures of Brachionus Nevada and Cayman were grown semi-continuously, fed live Rhodmonas baltica algae, and cultures were diluted daily (20% day−1). The enrichment diets Marol E (triacylglycerol-rich) and PL (cod roe emulsion, phospholipid-rich) were added to both cultures (Nevada 40 ng ind−1 day−1 and Cayman 25 ng ind−1 day−1, respectively). After 4 days of enrichment (dilution rate: 20% day−1), Cayman rotifers–fed Marol E had comparable (P > 0.05) quantitative amounts of DHA (docosahexaenoic acid, 22:6n-3) to natural harvested copepods, which were analysed for comparison, and the Cayman strain had generally more efficient incorporation of HUFAs (highly unsaturated fatty acids) than the Nevada strain. However, the percentage DHA of the total fatty acids in copepods were much higher than in enriched rotifers (P

The objectives of the study were to examine if early cod larvae could incorporate long-chain highly unsaturated fatty acids like docosahexaenoic acid (22:6 n−3, DHA) from dietary phospholipids (PL) more efficiently than from dietary triacylglycerides (TAG), and to investigate if because of this, PL from copepods is a better DHA source than PL from rotifers for larval cod. Two diets of Brachionus plicatilis Nevada were compared in one first feeding experiment. Their lipid composition was similar, but %DHA of PL fatty acids was different; 2.1% for Brachionus-Low and 9.4% for Brachionus-High. A second experiment compared the Brachionus-High diet (9.4% DHA in PL) and cultivated nauplii of Acartia tonsa that contained 30% DHA of PL-fatty acids. The total lipid per fry dry weight (DW) and the lipid class composition were similar for all larval groups at 17days post-hatching (dph). The %DHA of total fatty acids in PL of newly hatched larvae (2dph) was 26%, and the values for fed groups varied from 17 to 30% DHA of PL fatty acids. The dominant fatty acids in the PL of larvae and PL of the diets were well correlated (r2=0.84, p

The importance of n-3 long chain polyunsaturated fatty acids (LC-PUFAs) for human health has received more focus the last decades, and the global consumption of n-3 LC-PUFA has increased. Seafood, the natural n-3 LC-PUFA source, is harvested beyond a sustainable capacity, and it is therefore imperative to develop alternative n-3 LC-PUFA sources for both eicosapentaenoic acid (EPA, 20:5n-3) and docosahexaenoic acid (DHA, 22:6n-3). Genera of algae such as Nannochloropsis, Schizochytrium, Isochrysis and Phaedactylum within the kingdom Chromista have received attention due to their ability to produce n-3 LC-PUFAs. Knowledge of LC-PUFA synthesis and its regulation in algae at the molecular level is fragmentary and represents a bottleneck for attempts to enhance the n-3 LC-PUFA levels for industrial production. In the present review, Phaeodactylum tricornutum has been used to exemplify the synthesis and compartmentalization of n-3 LC-PUFAs. Based on recent transcriptome data a co-expression network of 106 genes involved in lipid metabolism has been created. Together with recent molecular biological and metabolic studies, a model pathway for n-3 LC-PUFA synthesis in P. tricornutum has been proposed, and is compared to industrialized species of Chromista. Limitations of the n-3 LC-PUFA synthesis by enzymes such as thioesterases, elongases, acyl-CoA synthetases and acyltransferases are discussed and metabolic bottlenecks are hypothesized such as the supply of the acetyl-CoA and NADPH. A future industrialization will depend on optimization of chemical compositions and increased biomass production, which can be achieved by exploitation of the physiological potential, by selective breeding and by genetic engineering. © 2013 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/3.0/).

The dietary requirement of phospholipid (PL) of fish larvae has been suggested to originate in an inefficient ability for de novo biosynthesis of PL based on dietary triacylglycerol (TAG). The main objective of the present study was to investigate whether cod larvae could synthesis PL from sn-2-monoacylglycerol (2-MAG) and glycerol precursors. A tube feeding method was used to deliver equal molar aliquots of 2-oleoyl-[1,2,3-(3)H]glycerol and [U-(14)C] glycerol together with bovine serum albumin (BSA) bound 16:0 (palmitic acid) and 22:6n-3 (docosahexaenoic acid, DHA), with or without choline chloride to the foregut of anesthetized cod larvae and thereafter monitoring the metabolism of these components in the larvae through 4 h following injection. Our results showed that both 2-MAG and glycerol precursors contributed to the de novo synthesis of phosphatidylcholine (PC) and the 2-MAG pathway predominated over the G-3-P (glycerol-3-phosphate) pathway in the synthesis of TAG and PC. The molecular ratio of PC/TAG obtained from the 2-MAG and the G-3-P pathways was 0.44-0.74 and 1.02-2.06 within the first hour of tube feeding, suggesting they might have comparable biosynthesis ability of PC and TAG under the conditions of the present study. Furthermore, supplementation of choline chloride significantly increased PC/TAG ratio (p < 0.05) for both pathways. However, further studies are needed to quantify the enzyme activity involved in the CDP-choline (cytidine diphosphate choline) pathway, and the function of choline either in simulating PC synthesis or TAG catabolism or both needs further investigation.