Clinical Efficacy, Analgesic Efficacy, and Effects of Cocktail Analgesic Regimens in Patients Undergoing Total Knee Arthroplasty: A Systematic Review and Meta-Analysis.

Objective: Multimodal cocktail analgesic injection (CAI) is widely used as an adjunct pain-reliever in the postoperative phase of patients undergoing total knee arthroplasty (TKA) due to intense postoperative pain accompanying the procedure leading to complications, thereby extending hospital stays. The aim of this study is to establish the clinical efficacy and effects of utilizing CAI regimens during the TKA procedure and the corresponding postoperative patient outcomes. Methods: A database search for pertinent articles literature search was performed in Embase, PubMed, Cochrane Library, Web of Science, and MEDLINE databases. RevMan version 5.4 was used to perform a meta-analysis on the included studies. Results: Data screening and selection produced 15 relevant articles that met the eligibility criteria of this study. The meta-analysis revealed insignificant difference between cocktail injected and control groups in VAS postoperative pain scores both at rest and during activity (OR 0.79, 95% CI 0.59 to 1.05; I2 = 0%; P = .93) and (OR 0.79, 95% CI 0.57 to 1.10; I2 = 0%; P = .75), respectively. Similarly, there was insignificant differences in postoperative knee flexion ROM, postoperative narcotic consumption, and length of stays between the two groups, (OR 1.20, 95% CI 1.03 to 1.40; P = .53), (OR 0.62, 95% CI 0.36 to 1.07; P = .09), and (OR 0.45, 95% CI 0.29 to 0.70; P = .21), respectively. However, the postoperative complications reveal statistical significance between the cocktail injected and the control group (OR 0.45, 95% CI 0.29 to 0.70; P = .004). Conclusion: It is concluded that CAI can play a crucial role in minimizing post-operative complications for patients undergoing TKA.

Correlation Between Central Vein Smoke Ultrasonography and Thrombus Elasticity Graphy in Severe Patients.

Objective: The objective of this study was to analyze the correlation between central vein smoke ultrasonography (CVSU) and thrombus elasticity graphy (TEG). Methods: A retrospective analysis was made on 300 severe patients with smoky echo changes (SECs) in the internal jugular and femoral veins who were admitted to the hospital from January 2021 to March 2022. According to the ultrasound results, all patients were divided into Group A (n = 75), Group B (n = 75), Group C (n = 75), and Group D (n = 75). TEG examination, ultrasound examination, routine coagulation test were received. The coagulation function and TEG index were compared and analyzed in each group, and their correlation was analyzed. Results: The trends of R value and K value of TEG index of patients in different groups were the same. The R value and K value in group D were the highest, followed by group C, and the lowest in group A; while those in groups C and D exhibited great differences with P < .05 to those in groups A and B. The PT, TT, APTT, and FIB of patients in groups C and D were much higher based on the values in groups A and B. R-value was positively correlated with APACHEII (0.678), TT (0.198), and APTT (0.187), and negatively associated with PT (-0.008), D-D (-0.315), and FDP (-0.298). K value presented a positive correlation with APACHEII (0.692) and TT (0.342) but a negative correlation with PT (-0.187), APTT (-0.053), D-D (-0.497), and FDP (-0.453). Positive correlations were observed between α and PT (0.198), APTT (0.046), D-D (0.602), and FDP (0.532), while negative correlations were found between α and APACHEII (-0.398) and TT (-0.315). MA was positively correlated with PT, D-D, and FDP but negatively with APACHEII, TT, and APTT. Conclusion: TEG parameters had an obvious correlation with the coagulation function test, which can effectively evaluate the CVSU in severe patients. The ultrasonic signs can be undertaken as clinical hypercoagulability detection indicators in severe patients and intervention indicators for early thrombosis prevention in the future, they can guide clinicians to make the best treatment plan for severe patients.

Bedside Ultrasound-guided Nasointestinal Tube Placement in Critically Ill Patients in Intensive Care Unit.

Objective: To verify the efficacy and safety of bedside ultrasound-guided nasointestinal tube (NIT) placement techniques in critically ill patients in the ICU. Methods: 100 Critically ill patients were selected and were randomly enrolled into a bedside ultrasound guidance (BUG) group (BUG guiding the NIT placement) and a traditional blind insertion (TBI) group, with 50 cases in both. The efficacy and safety of these tube placements were compared. Results: The success rate of intubation in the BUG group (74%) was higher than that in the TBI group (44%). The proportion of patients in the BUG group who had catheterization sites in the intestine (72%) was higher than that in the TBI group (46%) (P < .05). The average number of tube insertions and mean time of successful intubation time in the BUG group was slightly higher than those in the TBI group [(1.22 ± 0.00) times vs. (1.20 ± 1.00) times and (24.40 ± 0.50) min vs. (20.72 ± 0.50) min) (P > .05) respectively]. Conclusions: Bedside ultrasound-guided nasojejunal tube has a good outcome in ICU patients with critical conditions, can improve the success rate of intubation, and has a certain safety.

In vitro gastrointestinal digestion of buckwheat (Fagopyrum esculentum Moench) protein: release and structural characteristics of novel bioactive peptides stimulating gut cholecystokinin secretion.

Satiety hormone cholecystokinin (CCK) plays a vital role in appetite inhibition. Its secretion is regulated by dietary components. The search for bioactive compounds that stimulate CCK secretion is currently an active area of research. The objective of this study was to evaluate the ability of buckwheat (Fagopyrum esculentum Moench) protein digest (BPD) to stimulate CCK secretion in vitro and in vivo and clarify the structural characteristics of peptides stimulating CCK secretion. BPD was prepared by an in vitro gastrointestinal digestion model. The relative molecular weight of BPD was <10 000 Da, and peptides with <3000 Da accounted for 70%. BPD was rich in essential amino acids Lys, Leu, and Val but lacked sulfur amino acids Met and Cys. It had a stimulatory effect on CCK secretion in vitro and in vivo. Chromatographic separation was performed to isolate peptide fractions involved in CCK secretion, and five novel CCK-releasing peptides including QFDLDD, PAFKEEHL, SFHFPI, IPPLFP, and RVTVQPDS were successfully identified. A sequence length range of 6-8 and marked hydrophobicity (18-28) were observed among the most CCK-releasing peptides. The present study demonstrated for the first time that BPD could stimulate CCK secretion and clarify the structural characteristics of bioactive peptides having CCK secretagogue activity in BPD.

Analysis of oil synthesis pathway in Cyperus esculentus tubers and identification of oleosin and caleosin genes.

The tubers of the widely distributed Cyperus esculentus are rich in oil, and therefore, the plant is considered to have a high utilization value in the vegetable oil industry. Oleosins and caleosins are lipid-associated proteins found in oil bodies of seeds; however oleosins and caleosins genes have not been identified in C. esculentus. In this study, we performed transcriptome sequencing and lipid metabolome analysis of C. esculentus tubers at four developmental stages to obtain the information on their genetic profile, expression trends, and metabolites in oil accumulation pathways. Overall, 120,881 non-redundant unigenes and 255 lipids were detected; 18 genes belonged to the acetyl-CoA carboxylase (ACC), malonyl-CoA:ACP transacylase (MCAT), ß-ketoacyl-ACP synthase (KAS), and fatty acyl-ACP thioesterase (FAT) gene families involved in fatty acid biosynthesis, and 16 genes belonged to the glycerol-3-phosphate acyltransferase (GPAT), diacylglycerol acyltransferase 3 (DGAT3), phospholipid:diacylglycerol acyltransferase (PDAT), FAD2, and lysophosphatidic acid acyltransferase (LPAAT) gene families playing important roles in triacylglycerol synthesis. We also identified 9 oleosin- and 21 caleosin-encoding genes in C. esculentus tubers. These results provide detailed information on the C. esculentus transcriptional and metabolic profiles, which can be used as reference for the development of strategies to increase oil content in C. esculentus tubers.

Genome-wide identification and expression analysis of diacylglycerol acyltransferase genes in soybean (Glycine max).

Background: Soybean (Glycine max) is a major protein and vegetable oil source. In plants, diacylglycerol acyltransferase (DGAT) can exert strong flux control, which is rate-limiting for triacylglycerol biosynthesis in seed oil formation. Methods: Here, we identified soybean DGAT genes via a bioinformatics method, thereby laying a solid foundation for further research on their function. Based on our bioinformatics analyses, including gene structure, protein domain characteristics, and phylogenetic analysis, 26 DGAT putative gene family members unevenly distributed on 12 of the 20 soybean chromosomes were identified and divided into the following four groups: DGAT1, DGAT2, WS/DGAT, and cytoplasmic DGAT. Results: The Ka/Ks ratio of most of these genes indicated a significant positive selection pressure. DGAT genes exhibited characteristic expression patterns in soybean tissues. The differences in the structure and expression of soybean DGAT genes revealed the diversity of their functions and the complexity of soybean fatty acid metabolism. Our findings provide important information for research on the fatty acid metabolism pathway in soybean. Furthermore, our results will help identify candidate genes for potential fatty acid-profile modifications to improve soybean seed oil content. Conclusions: This is the first time that in silico studies have been used to report the genomic and proteomic characteristics of DGAT in soybean and the effect of its specific expression on organs, age, and stages.

Key knowledge gaps for One Health approach to mitigate nanoplastic risks.

There are increasing concerns over the threat of nanoplastics to environmental and human health. However, multidisciplinary barriers persist between the communities assessing the risks to environmental and human health. As a result, the hazards and risks of nanoplastics remain uncertain. Here, we identify key knowledge gaps by evaluating the exposure of nanoplastics in the environment, assessing their bio-nano interactions, and examining their potential risks to humans and the environment. We suggest considering nanoplastics a complex and dynamic mixture of polymers, additives, and contaminants, with interconnected risks to environmental and human health. We call for comprehensive integration of One Health approach to produce robust multidisciplinary evidence to nanoplastics threats at the planetary level. Although there are many challenges, this holistic approach incorporates the relevance of environmental exposure and multi-sectoral responses, which provide the opportunity to identify the risk mitigation strategies of nanoplastics to build resilient health systems.

Quantitative proteomic and lipidomics analyses of high oil content GmDGAT1-2 transgenic soybean illustrate the regulatory mechanism of lipoxygenase and oleosin.

KEY MESSAGE: Proteomic and lipidomics analyses of WT and GmDGAT1-2 transgenic soybeans showed that GmDGAT1-2 over-expression induced lipoxygenase down-regulatation and oleoin up-regulatation, which significantly changed the compositions and total fatty acid. The main goal of soybean breeding is to increase the oil content. Diacylglycerol acyltransferase (DGAT) is a key rate-limiting enzyme in fatty acid metabolism and may regulate oil content. Herein, 10 GmDGAT genes were isolated from soybean and transferred into wild-type (WT) Arabidopsis. The total fatty acid was 1.2 times higher in T3 GmDGAT1-2 transgenic Arabidopsis seeds than in WT. Therefore, GmDGAT1-2 was transferred into WT soybean (JACK), and four T3 transgenic soybean lines were obtained. The results of high-performance gas chromatography and Soxhlet extractor showed that, compared with those of JACK, oleic acid (18:1), and total fatty acid levels in transgenic soybean plants were much higher, but linoleic acid (18:2) was lower than WT. Palmitic acid (16:0), stearic acid (18:0), and linolenic acid (18:3) were not significantly different. For mechanistic studies, 436 differentially expressed proteins (DEPs) and 180 differentially expressed metabolites (DEMs) were identified between WT (JACK) and transgenic soybean pods using proteomic and lipidomics analyses. Four lipoxygenase proteins were down-regulated in linoleic acid metabolism while four oleosin proteins were up-regulated in the final oil formation. The results showed an increase in the total fatty acid and 18:1 composition, and a decrease in the 18:2 composition of fatty acid. Our study brings new insights into soybean genetic transformation and the deep study of molecular mechanism that changes the total fatty acid, 18:1, and 18:2 compositions in GmDGAT1-2 transgenic soybean.

Bergenin Monohydrate Attenuates Inflammatory Response via MAPK and NF-κB Pathways Against Klebsiella pneumonia Infection.

Background: Klebsiella pneumonia has emerged as a critical pathogen causing severe clinical problems, such as pneumonia and sepsis. Meanwhile, intensified drug resistance induced by antibiotic therapy necessitates discovering novel and active molecules from Traditional Chinese Medicine (TCM) for treatment. Methods and results: In this study, the isolated Bergenin monohydrate showed an anti-inflammatory effect in Klebsiella-infected mice. We initially investigated the anti-inflammatory effects and cytoprotection against oxidative stress in vitro and in vivo. Interestingly, a specific dose of Bm can effectively ameliorate lung injury and suppress the expression of inflammatory cytokines such as TNF-α, IL-6, IL-1ß and PEG2. Moreover, Bm was also shown to reduced the levels of MPO, MDA and increased SOD and GSH activities. Moreover, we assessed the intracellular signaling molecules including p38, ERK, JNK, IκB, NF-κB-p65 by western blotting and verified through MAPK and NF-κB pathways inhibition experiments. These results reveal that Bm executed its effects via the classical MAPK signaling pathway and NF-κB pathway. Conclusion: Given its underlying anti-inflammatory effect, Bm may be used as a promising therapeutic against Klebsiella-induced infection, thus providing a benefit for the future clinical therapy of pneumonia and medicine design.

Potassium supplementation blunts the effects of high salt intake on serum retinol-binding protein 4 levels in healthy individuals.

AIMS/INTRODUCTION: Excessive dietary salt or low potassium intakes are strongly correlated with insulin resistance (IR) and type 2 diabetes mellitus. In epidemiological and experimental studies, increased serum retinol-binding protein 4 (RBP4) contributes to the pathogenesis of type 2 diabetes mellitus. Herein, we hypothesized that RBP4 might be an adipocyte-derived "signal" that plays the crucial role in salt-related insulin resistance or type 2 diabetes mellitus. This study aimed to assess whether salt consumption and potassium supplementation influence serum RBP4 levels in healthy individuals. MATERIALS AND METHODS: A total of 42 participants (aged 25-50 years) in a rural area of Northern China were successively provided normal (3 days at baseline), low-salt (7 days; 3 g/day NaCl) and high-salt (7 days; 18 g/day) diets, and a high-salt diet with potassium additive (7 days; 18 g/day NaCl and 4.5 g/day KCl). Urinary sodium and potassium were measured to ensure compliance to dietary intervention. Then, RBP4 levels were evaluated by enzyme-linked immunosorbent assay. RESULTS: High salt intake significantly raised serum RBP4 levels in healthy participants (17.5 ± 0.68 vs 28.6 ± 1.02 µg/mL). This phenomenon was abrogated by potassium supplementation (28.6 ± 1.02 vs 17.6 ± 0.88 µg/mL). In addition, RBP4 levels presented positive (r = 0.528, P < 0.01) and negative (r = -0.506, P < 0.01) associations with 24-h urinary sodium- and potassium excretion levels. CONCLUSIONS: RBP4 synthesis is motivated by high salt intake and revoked by potassium supplementation. Our pioneer work has contributed to the present understanding of salt-induced insulin resistance or type 2 diabetes mellitus.

Protocatechuic aldehyde from Salvia miltiorrhiza exhibits an anti-inflammatory effect through inhibiting MAPK signalling pathway.

BACKGROUND: The aerial parts of Salvia miltiorrhiza, which was considered to be the waste part and discarded during the root harvest, is rich in protocatechuic aldehyde (PAI). This study investigated the health-promoting effects of extracts and PAI from the aerial parts of Salvia miltiorrhiza, including its anti-inflammatory effects and the underlying mechanisms of action in vitro and in vivo. METHOD: Purification of the sample paste of Salvia miltiorrhiza was accomplished using HPLC analysis. TheMTT (Methylthiazolyldiphenyl-tetrazolium bromide) assay was employed to determine the cell viability. The production of inflammatory factors was detected by ELISA assays. The histopathological analysis was used to analyse the lungs and livers of mice treated with PAI. Western blot was performed to reveal the mechanism of PAI in anti-inflammatory. RESULTS: The extracts and PAI from the aerial parts of Salvia miltiorrhiza inhibited TNF-α, IL-6 production and promoted the production of IL-10 in vivo in mice and in vitro in the macrophage cell line RAW264.7. NF-κB and MAPKs kinase phosphorylation were also suppressed by PAI in vivo and in vitro, indicating that PAI exhibited an anti-inflammatory effect. CONCLUSION: These findings suggest that the aerial parts of Salvia miltiorrhiza extract may serve as potential protective agents for inflammatory.

Effects of multiple N, P, and K fertilizer combinations on adzuki bean (Vigna angularis) yield in a semi-arid region of northeastern China.

Nitrogen (N), phosphorus (P), and potassium (K) exert various effects on adzuki bean yields. Our research was conducted in a semi-arid area, and four test sites were established in environments that have chernozem or sandy loam soils. During a five-year period, the effects of N, P, and K fertilizers on yield were comprehensively investigated in field trials (2014-2016) and for model-implementation trials (2017-2018), with models established prior to the latter. In the field trials, 23 treatments comprising different N, P, and K combinations significantly affected both yield and yield components, and regression analysis indicated that the experimental results were suitable for model establishment. The model subsequently demonstrated that the yield and the yield components were more sensitive to N and K fertilizer than to P fertilizer. Moreover, the yield and yield components increased. These yield increases were intense in response to the 0.5 to 1.34 levels in terms of the single effects; interaction effects; and the effects of combinations of N, P, and K fertilizers. Moreover, the effects of combinations of N, P, and K fertilizers were more significant on yield than were the single or interaction effects of N, P, and K fertilizers. The optimal fertilizer combination that resulted in high yields (≥1941.53 kg ha-1) comprised 57.23-68.43 kg ha-1 N, 36.04-47.32 kg ha-1 P2O5 and 50.29-61.27 kg ha-1 K2O. The fertilizer combination that resulted in the maximum yield was 62.98 kg ha-1 N, 47.04 kg ha-1 P2O5 and 59.95 kg ha-1 K2O (N:P2O5:K2O = 1:0.75:0.95), which produced the model-expected yield in trials at multiple sites. An economical fertilizer combination was determined on the basis of the best fertilizer measures in consideration of the cost of fertilizer and seed; this combination achieved yields of 2236.17 kg ha-1, the profit was 15,653.16 Yuan ha-1, and the corresponding rates were 57.60 kg ha-1 N, 47.03 kg ha-1 P2O5, and 31.64 kg ha-1 K2O (N:P2O5:K2O = 1:0.82:0.55).

Nitrogen, phosphorus, and potassium fertilization to achieve expected yield and improve yield components of mung bean.

Mung bean (Vigna radiata L.) is an important edible bean in the human diet worldwide. However, its growth, development, and yield may be restricted or limited by insufficient or unbalanced nitrogen (N), phosphorus (P), and potassium (K) fertilization. Despite this, there are few long-term studies of the effects of varying levels of N, P, and K combined fertilizers and the optimal fertilization for improving mung bean yield and quality. This study was conducted to optimize the fertilization strategies for high yield and to improve yield components (pods per plant, seeds per pod, and 100-seed weight) in the Bailv9 mung bean cultivar, 23 treatments were tested in 2013-2015, using a three-factor (N, P, and K fertilizers), five-level quadratic orthogonal rotation combination design. Our studies showed that, the N, P, and K fertilizers significantly influenced the pods per plant and yield, which increased and then decreased with the increasing N, P, and K fertilizers. The 100-seed weight was significantly affected by the N and P fertilization, and it was increased consistently with the increasing N fertilizer, and decreased significantly with the increasing P fertilizer. Whereas, the seeds per pod significantly decreased with the increasing N and K fertilizers, and the P fertilizer had no significant effect on it. The NP interaction had a significant effect on yield and pods per plant at high N levels, while the NK interaction had a significant but opposite effect on yield at low N levels. The optimal fertilization conditions to obtain yield >2,141.69 kg ha-1 were 34.38-42.62 kg ha-1 N, 17.55-21.70 kg ha-1 P2O5, and 53.23-67.29 kg ha-1 K2O. Moreover, the optimal N, P, and K fertilization interval to achieve pods per plant > 23.41 and the optimal N fertilization to achieve a 100-seed weight > 6.58 g intersected with the interval for yield, but the seeds per pod did not. The fertilizer ratio for the maximum yield was N:P2O5:K2O = 1:0.5:1.59. Following three years experimentation, the optimal fertilization measures were validated in 2016-2017, the results indicated that yield increased by 19.6% than that obtained using conventional fertilization. The results of this study provide a theoretical basis and technical guidance for high-yield mung bean cultivation using the optimal fertilization measures.

Structure-affinity relationship of the interaction between phenolic acids and their derivatives and ß-lactoglobulin and effect on antioxidant activity.

In this study, 71 phenolic acids and their derivatives were used to investigate the structure-affinity relationship of ß-lactoglobulin binding, and the effect of this interaction on antioxidant activity. Based on a fluorescence quenching method, an improved mathematical model was adopted to calculate the binding constants, with a correction for the inner-filter effect. Hydroxylation at the 3-position increased the affinity of the phenolic acids for ß-lactoglobulin, while hydroxylation at the 2- or 4-positions had a negative effect. Complete methylation of all hydroxy groups, except at the 3-position, enhanced the binding affinity. Replacing the hydroxy groups with methyl groups at the 2-position also had a positive effect. Hydrogen bonding was one of the binding forces for the interaction. The antioxidant activity of phenolic acid-ß-lactoglobulin complexes was higher than that of phenolic acids alone. These findings provide an understanding of the structure-activity relationship of the interaction between ß-lactoglobulin and phenolic acids.

Selenium Deficiency-Induced Apoptosis of Chick Embryonic Vascular Smooth Muscle Cells and Correlations with 25 Selenoproteins.

Selenium deficiency is the major cause of exudative diathesis in chicks. Subcutaneous hemorrhage is one of the typical symptoms of the disease. However, the reason for the occurrence of blood exudation remains unknown. In the present study, the vascular smooth muscle cells (VSMCs) were isolated from 17-day-old broiler chick embryos. Cell viability, cell apoptosis, and intracellular reactive oxygen species level under different concentrations of selenium (0-0.9 µM) were investigated. The mRNA expression levels of 25 selenoproteins and apoptosis-related genes (p53, CytC, Caspase-3, Caspase-8, Bcl-2, and Bax) were also measured. Selenium deficiency significantly decreased cell viability and increased cell apoptosis (p < 0.05). Supplementation with selenium could alleviate these changes. In general, at all levels of selenium addition, Gpx1, Gpx3, Gpx4, SepW1, and Sep15 mRNAs were all highly expressed in VSMCs, whereas Gpx2, Dio1, SepN1, SelO, and SelPb were at lower levels. There was a high correlation between Gpx2, Gpx3, Gpx4, Dio1, Txnrd1, Txnrd2, and Txnrd3 gene expression. Additionally, Gpx3, Gpx4, Dio1, Txnrd1, Txnrd2, Txnrd3, SelS, and SelPb showed a strong negative correlation with pro-apoptotic gene Caspase-3 as well as a strong positive correlation with anti-apoptotic gene Bcl-2, especially SelI (r = 0.913 and r = 0.929, p < 0.01). These results suggest that selenium deficiency could induce VSMC apoptosis, and several selenoproteins may be involved in the development of apoptosis. Our findings provide information on the molecular mechanism of vascular injury by selenium deficiency.

Effects of a Tripeptide Iron on Iron-Deficiency Anemia in Rats.

This study aims to investigate the effects of a tripeptide iron (REE-Fe) on iron-deficiency anemia rats. Sprague-Dawley rats were randomly divided into seven groups: a normal control group, an iron-deficiency control group, and iron-deficiency groups treated with ferrous sulfate (FeSO4), ferrous glycinate (Fe-Gly), or REE-Fe at low-, medium-, or high-dose groups. The rats in the iron-deficiency groups were fed on an iron-deficient diet to establish iron-deficiency anemia (IDA) model. After the model established, different iron supplements were given to the rats once a day by intragastric administration for 21 days. The results showed that REE-Fe had effective restorative action returning body weight, organ coefficients, and hematological parameters in IDA rats to normal level. In addition, comparing with FeSO4 or Fe-Gly, high-dose REE-Fe was more effective on improving the levels of renal coefficient, total iron-binding capacity, and transferrin. Furthermore, the liver hepcidin messenger RNA (mRNA) expression in the high-dose group was significantly higher (p < 0.05) than that in the FeSO4 or Fe-Gly group and showed no significant difference (p > 0.05) with the normal control group. The findings suggest that REE-Fe is an effective source of iron supplement for IDA rats and might be exploited as a new iron fortifier.

Modulation of gut microbiota by polyphenols from adlay (Coix lacryma-jobi L. var. ma-yuen Stapf.) in rats fed a high-cholesterol diet.

This study aimed to evaluate the beneficial effects of polyphenol extract of adlay (Coix lacryma-jobi L. var. ma-yuen Stapf.) (PEA) on gut microbiota in rats fed a high-cholesterol diet (HCD). Rats were fed HCD containing 1% cholesterol (w/w), with or without a daily intragastric supplement of 200 mg/kg body weight PEA. Results showed that PEA significantly ameliorated increases in serum cholesterol and low-density lipoprotein cholesterol values and significantly restored high-density lipoprotein cholesterol values. The HCD-induced imbalance of gut microflora was modulated by the consumption of PEA. Most bacterial strains influenced by PEA are related to host lipid metabolism. The abundances of one Erysipelotrichales strains and two Clostridia strains were lower in the PEA group than in the control. Phenolic compounds in PEA were identified by HPLC. The findings indicate that PEA may be a useful dietary supplement in the treatment of elevated cholesterol levels and the imbalanced gut microbial ecology.

A R2R3-MYB transcription factor, GmMYB12B2, affects the expression levels of flavonoid biosynthesis genes encoding key enzymes in transgenic Arabidopsis plants.

Isoflavones play diverse roles in plant-microbe interactions and are potentially important for human nutrition and health. To study the regulation of isoflavonoid synthesis in soybean, the R2R3-MYB transcription factor GmMYB12B2 was isolated and characterized. Yeast expression experiments demonstrated that GmMYB12B2 showed transcriptional activity. GmMYB12B2 was localized in the nucleus when it was transiently expressed in onion epidermal cells. Real-time quantitative PCR analysis revealed that GmMYB12B2 transcription was increased in roots and mature seeds compared with other organs. The gene expression level in immature embryos was consistent with the accumulation of isoflavones. CHS8 is a key enzyme in plant flavonoid biosynthesis. Transient expression experiments in soybean calli demonstrated that CHS8 was regulated by GmMYB12B2 and produced more fluorescence. The expression levels of some key enzymes in flavonoid biosynthesis were examined in transgenic Arabidopsis lines. The results showed that the expression levels of PAL1, CHS and FLS in transgenic plants were significantly higher than those in wild type plants. However, the expression level of DFR was lower, and the expression levels of CHI, F3H and F3'H were the same in all lines. GmMYB12B2 expression caused a constitutive increase in the accumulation of flavonoids in transgenic Arabidopsis lines compared with wild type plants.

Isolation and molecular characterization of GmERF7, a soybean ethylene-response factor that increases salt stress tolerance in tobacco.

Ethylene-response factors (ERFs) play an important role in regulating gene expression in plant responses to biotic and abiotic stresses. In this study, a new ERF transcription factor, GmERF7, was isolated from soybean. Sequence analysis showed that GmERF7 contained an AP2/ERF domain with 58 amino acids, two putative nuclear localization signal (NLS) domains, an acidic amino acid-rich transcriptional activation domain and a conserved N-terminal motif [MCGGAI(I/L)]. The expression of GmERF7 was induced by drought, salt, methyl jasmonate (MeJA), ethylene (ETH) and abscisic acid (ABA) treatments. However, the expression of GmERF7 decreased under cold treatment. GmERF7 localized to the nucleus when transiently expressed in onion epidermal cells. Furthermore, GmERF7 protein bound to the GCC-box element in vitro and activated the expression of the ß-glucuronidase (GUS) reporter gene in tobacco leaves. Activities of GmERF7 promoter (GmERF7P) upregulated in tobacco leaves with 10h drought, salt and ETH treatments. However, activities of GmERF7P decreased with 10h cold and ABA treatments. Overexpression of GmERF7 in tobacco plants led to higher levels of chlorophyll and soluble carbohydrates and a lower level of malondialdehyde compared with wild-type tobacco plants under salt stress conditions, which indicated that GmERF7 enhanced salt tolerance in transgenic plants.

Expression of Chlamydophila psittaci MOMP heat-labile toxin B subunit fusion gene in transgenic rice.

A DNA fragment encoding the MOMP gene of Chlamydophila psittaci was fused to the heat-labile toxin B subunit gene (LTB-MOMP) and transferred into rice callus by Agrobacterium tumefaciens-mediated transformation. The LTB-MOMP fusion gene was detected in genomic DNA from transformed rice leaves by Southern blot and RT-PCR amplification. Synthesis and assembly of the LTB-MOMP fusion protein into pentamers was detected in transformed leaf extracts by immunoblot analysis. Binding of the pentamers to intestinal epithelial cell membrane glycolipid receptors was quantified by GM1-ganglioside enzyme-linked immunosorbent assay (GM1-ELISA). The ELISA results indicated that LTB-MOMP fusion protein made up 0.0033-0.0054% of the total soluble leaf protein. Meanwhile, this suggested that the fusion protein retained both its native antigenicity and the ability to form pentamers.