Tea consumption and risk of bone health: an updated systematic review and meta-analysis.

INTRODUCTION: Current research evaluating the association between tea consumption and bone health still has inconsistent findings. MATERIALS AND METHODS: The electronic databases of Embase, PubMed, Scopus, and Web of Science were systematically searched from inception until December 2022 to identify eligible studies. The calculation of summary relative risks (RRs) and 95% confidence intervals (CIs) was carried out using random-effects models. I2 statistics and Forest plots were used to assess the heterogeneity of RR values across studies. RESULTS: The pooled relative risks for bone health-related outcomes of interest among tea drinkers, compared to non-drinkers, were 0.910 (95% confidence interval 0.845 to 0.980) for fractures, based on 20 studies, 0.332 (0.207-0.457) for BMD (13 studies), 0.800 (0.674-0.950) for osteoporosis (10 studies), and 1.006 (0.876-1.156) for osteopenia (5 studies). Subgroup analysis of locations showed that the pooled relative risks were 0.903 (0.844-0.966) for the hip, 0.735 (0.586-0.922) for the femur, 0.776 (0.610-0.988) for the lumbar, 0.980 (0.942-1.021) for the forearm and wrist, 0.804 (0.567-1.139) for the phalanges, and 0.612 (0.468-0.800) for Ward's triangle. One-stage dose-response analysis revealed that individuals who consumed less than 4.5 cups of tea per day had a lower risk of bone health-related outcomes than those who did not consume tea, with statistically significant results. CONCLUSION: There is an association between tea consumption and a reduced risk of fractures, osteoporosis, hip, femur, and lumbar, as well as increased BMD.

Effects of two fillers and process conditions on the water treatment efficiency of a continuous packed bed biofilm reactor.

This study evaluated the treatment efficiency of two selected fillers and their combination for improving the water quality of aquaculture wastewater using a packed bed biofilm reactor (PBBR) under various process conditions. The fillers used were nanosheet (NS), activated carbon (AC), and a combination of both. The results indicated that the use of combined fillers and the hydraulic retention time (HRT) of 4 h significantly enhanced water quality in the PBBR. The removal rates of chemical oxygen demand, NO2-─N, total suspended solids（TSS）, and chlorophyll a were 63.55%, 74.25%, 62.75%, and 92.85%, respectively. The microbiota analysis revealed that the presence of NS increased the abundance of microbial phyla associated with nitrogen removal, such as Nitrospirae and Proteobacteria. The difference between the M1 and M2 communities was minimal. Additionally, the microbiota in different PBBR samples displayed similar preferences for carbon sources, and carbohydrates and amino acids were the most commonly utilized carbon sources by microbiota. These results indicated that the combination of NS and AC fillers in a PBBR effectively enhanced the treatment efficiency of aquaculture wastewater when operated at an HRT of 4 h. The findings provide valuable insights into optimizing the design of aquaculture wastewater treatment systems.

Advanced perioperative assessment of neurological function in acute Stanford A aortic dissection.

OBJECTIVE: Acute Stanford Type A aortic dissection (AAAD) is a critical condition in vascular surgery, and total aortic arch replacement surgery is the preferred method to save patients' lives. In recent years, as clinical research has advanced, there has been a growing realization of the close association between poor postoperative outcomes in patients and neurological functional deficits. Neurological function monitoring is a medical technique used to evaluate and monitor the functional status of the nervous system. METHODS: This monitoring involves the assessment of various aspects of the nervous system, including but not limited to nerve conduction velocity, neuromuscular function, electroencephalographic activity, and sensory nerve transmission. Neurological function monitoring has broad clinical applications and can be used to diagnose and monitor many neurological disorders, helping physicians understand patients' neurological functional status and guide treatment plans. During the postoperative recovery process, neurological function monitoring can assist physicians in assessing the potential impact of surgery on the nervous system and monitor the recovery of patients' neurological function. RESULTS: Studies have shown that neurological function monitoring holds promise in predicting neurological functional prognosis and interventions for patients with aortic dissection. CONCLUSION: Therefore, the primary objective of this study is to evaluate the effectiveness and reliability of various intraoperative neurological monitoring techniques, neuroimaging examinations, and biomarkers in predicting and assessing postoperative neurological outcomes in patients undergoing AAAD surgery.

Coffee, Green Tea Intake, and the Risk of Hepatocellular Carcinoma: A Systematic Review and Meta-Analysis of Observational Studies.

Several studies suggest an inverse relationship between coffee intake and risk of hepatocellular carcinoma (HCC), but the association between green tea intake and the risk of HCC is still inconclusive. We performed a meta-analysis of observational studies to clarify the association. We identified eligible studies published from January 1, 1992, to February 28, 2022, by searching PubMed, Web of Science, and EMBASE. A total of 32 studies were included in the meta-analysis. Among them, 21 studies involving 2,492,625 participants and 5980 cases of HCC reported coffee intake, 18 studies involving 1,481,647 participants and 6985 cases of HCC reported green tea intake, and seven studies reported both coffee intake and green tea intake. The results showed that a higher coffee (RR = 0.53; 95% CI: 0.47-0.59; I2 = 0.0%; Pheterogeneity = 0.634) or green tea (RR = 0.80; 95% CI: 0.67-0.95; I2 = 72.30%; Pheterogeneity < 0.001) intake may be associated with a lower risk of HCC. The same results were observed in both cohort and case-control subgroups. Our findings suggest that drinking coffee or green tea may be a potentially effective approach for the prevention or mitigation of HCC, but this still needs to be confirmed by further well-designed observational studies and clinical experimental research.

Complete genome analysis of Pseudomonas furukawaii ZS1 isolated from grass carp (Ctenopharyngodon idellus) culture water.

Pseudomonas furukawaii ZS1, isolated from grass carp (Ctenopharyngodon idellus) culture water, exhibits efficient aerobic nitrate reduction without nitrite accumulation; however, the molecular pathway underlying this aerobic nitrate reduction remains unclear. In this study, we constructed a complete genome map of P. furukawaii ZS1 and performed a comparative genomic analysis with a reference strain. The results showed that P. furukawaii ZS1 genome was 6 026 050 bp in size and contained 5427 predicted protein-coding sequences. The genome contained all the necessary genes for the dissimilatory nitrate reduction to ammonia pathway but lacked those for the assimilatory nitrate reduction pathway; additionally, genes that convert ammonia to organic nitrogen were also identified. The presence of putative genes associated with the nitrogen and oxidative phosphorylation pathways implied that ZS1 can perform respiration and nitrate reduction simultaneously under aerobic conditions, so that nitrite is rapidly consumed for detoxication by denitrification. The aim of this study is to indicate the great potential of strain ZS1 for future full-scale applications in aquaculture. This work provided insights at the molecular level on the nitrogen metabolic pathways in Pseudomonas species. The understanding of nitrogen metabolic pathways also provides significant molecular information for further Pseudomonas species modification and development.

Quorum-quenching potential of recombinant PvdQ-engineered bacteria for biofilm formation.

Quorum sensing (QS) is a core mechanism for bacteria to regulate biofilm formation, and therefore, QS inhibition or quorum quenching (QQ) is used as an effective and economically feasible strategy against biofilms. In this study, the PvdQ gene encoding AHL acylase was introduced into Escherichia coli (DE3), and a PvdQ-engineered bacterium with highly efficient QQ activity was obtained and used to inhibit biofilm formation. Gene sequencing and western blot analysis showed that the recombinant pET-PvdQ strain was successfully constructed. The color reaction of Agrobacterium tumefaciens A136 indicated that PvdQ engineering bacteria had shown strong AHL signal molecule quenching activity and significantly inhibited the adhesion (motility) of Pseudomonas aeruginosa and biofilm formation of activated sludge bacteria in Membrane Bio-Reactor (MBR; inhibition rate 51-85%, p < 0.05). In addition, qRT-PCR testing revealed that recombinant PvdQ acylase significantly reduced the transcription level of QS biofilm formation-related genes (cdrA, pqsA, and lasR; p < 0.05). In this study, QQ genetically engineered bacteria enhanced by genetic engineering could effectively inhibit the QS signal transduction mechanism and have the potential to control biofilm formation of pathogenic bacteria in the aquaculture environment, providing an environmentally friendly and alternative antibiotic strategy to suppress biofilm contamination.

Application of the Powell algorithm for estimating optical properties of a semitransparent medium based on time-domain information.

Accurate estimation of the optical properties of a semitransparent medium is crucial in various engineering applications. This study introduces the Powell algorithm to estimate the optical properties of a 2D semitransparent slab. The time-domain radiative transfer equation is solved using the discrete ordinate method. The radiative intensity on the medium's surface serves as the measurement signal for the inverse analysis. The results demonstrate that the Powell algorithm accurately estimates the absorption coefficient, scattering coefficient, and scattering asymmetry factor. For simultaneous reconstruction of these three parameters, it is recommended to use eight signal detectors on both the left and right sides of the medium. Even when the standard measurement error is increased to 15%, the relative errors for these three parameters remain low, at 1.87%, 1.379%, and 0.194%.

LmFKBP24 interacts with LmEaster to inhibit the antifungal immunity of Locusta migratoria.

Locusta migratoria is one of the most destructive pests that threaten crop growth and food production security in China. Metarhizium anisopliae has been widely used to control locusts around the world. Previous laboratory studies have revealed that LmFKBP24 is significantly upregulated after M. anisopliae infection, suggesting that it may play a role in immune regulation, yet the mechanism remains largely unknown. To gain further insight, we conducted an RNA interference (RNAi) study to investigate the function of LmFKBP24 in the regulation of antifungal immunity and analyzed the expression patterns of immune-induced genes. Our research revealed that LmFKBP24 is activated and upregulated when locusts are infected by M. anisopliae, and it inhibits the expression of antimicrobial peptide (AMP) defensin in the downstream of Toll pathway by combining with LmEaster rather than LmCyPA, thus exerting an immunosuppressive effect. To further investigate this, we conducted yeast two-hybrid (Y2H) and pull down assays to identify the proteins interacting with LmFKBP24. Our results provided compelling evidence for revealing the immune mechanism of L. migratoria and uncovered an innovative target for the development of new biological pesticides. Furthermore, our research indicates that LmFKBP24 interacts with LmEaster through its intact structure, providing a strong foundation for further exploration.

The Conserved Cysteine-Rich Secretory Protein MaCFEM85 Interacts with MsWAK16 to Activate Plant Defenses.

Metarhizium anisopliae is an entomopathogenic fungus which may enhance plant growth and resistance when acting as an endophyte in host plants. However, little is known about the protein interactions nor their activating mechanisms. Common in fungal extracellular membrane (CFEM) proteins have been identified as plant immune regulators that suppress or activate plant resistance responses. Here, we identified a CFEM domain-containing protein, MaCFEM85, which was mainly localized in the plasma membrane. Yeast two-hybrid (Y2H), glutathione-S-transferase (GST) pull-down, and bimolecular fluorescence complementation assays demonstrated that MaCFEM85 interacted with the extracellular domain of a Medicago sativa (alfalfa) membrane protein, MsWAK16. Gene expression analyses showed that MaCFEM85 and MsWAK16 were significantly upregulated in M. anisopliae and M. sativa, respectively, from 12 to 60 h after co-inoculation. Additional yeast two-hybrid assays and amino acid site-specific mutation indicated that the CFEM domain and 52th cysteine specifically were required for the interaction of MaCFEM85 with MsWAK16. Defense function assays showed that JA was up-regulated, but Botrytis cinerea lesion size and Myzus persicae reproduction were suppressed by transient expression of MaCFEM85 and MsWAK16 in the model host plant Nicotiana benthamiana. Collectively, these results provide novel insights into the molecular mechanisms underlying interactions of M. anisopliae with host plants.

Hif1α/Dhrs3a Pathway Participates in Lipid Droplet Accumulation via Retinol and Ppar-γ in Fish Hepatocytes.

Excessive hepatic lipid accumulation is a common phenomenon in cultured fish; however, its underlying mechanisms are poorly understood. Lipid droplet (LD)-related proteins play vital roles in LD accumulation. Herein, using a zebrafish liver cell line (ZFL), we show that LD accumulation is accompanied by differential expression of seven LD-annotated genes, among which the expression of dehydrogenase/reductase (SDR family) member 3 a/b (dhrs3a/b) increased synchronously. RNAi-mediated knockdown of dhrs3a delayed LD accumulation and downregulated the mRNA expression of peroxisome proliferator-activated receptor gamma (pparg) in cells incubated with fatty acids. Notably, Dhrs3 catalyzed retinene to retinol, the content of which increased in LD-enriched cells. The addition of exogenous retinyl acetate maintained LD accumulation only in cells incubated in a lipid-rich medium. Correspondingly, exogenous retinyl acetate significantly increased pparg mRNA expression levels and altered the lipidome of the cells by increasing the phosphatidylcholine and triacylglycerol contents and decreasing the cardiolipin, phosphatidylinositol, and phosphatidylserine contents. Administration of LW6, an hypoxia-inducible factor 1α (HIF1α) inhibitor, reduced the size and number of LDs in ZFL cells and attenuated hif1αa, hif1αb, dhrs3a, and pparg mRNA expression levels. We propose that the Hif-1α/Dhrs3a pathway participates in LD accumulation in hepatocytes, which induces retinol formation and the Ppar-γ pathway.

Meat Intake and the Risk of Hepatocellular Carcinoma: A Meta-Analysis of Observational Studies.

The association between meat intake and hepatocellular carcinoma (HCC) risk is still unclear. We conducted a meta-analysis with observational studies to clarify this relationship. A total of 17 studies involving 2,915,680 participants and 4,953 cases of HCC were included in the meta-analysis. Ten studies reported red meat intake, nine reported white meat intake, nine reported fish intake, seven reported processed meat intake, and five reported total meat intake. The results showed that the consumption of red meat (relative risk [RR] = 1.04; 95% confidence interval [CI]: 0.91-1.18; I2=50.50%; P = 0.033) and total meat intake (RR = 1.01; 95% CI: 0.90-1.13; I2 = 15.50%; P = 0.316) were not associated with risk of HCC. However, a higher dietary intake of processed meat (RR = 1.20; 95% CI: 1.02-1.41; I2 = 26.30%; P = 0.228) may increase the risk of HCC. In contrast, the intake of white meat (RR = 0.76; 95% CI: 0.63-0.92; I2 = 68.30%; P = 0.001) and fish (RR = 0.91; 95% CI: 0.86-0.96; I2 =40.90%; P = 0.095) were inversely associated with risk of HCC. Our findings suggest that dietary intervention may be an effective approach to preventing HCC. These need to be verified with further well-designed observational studies and experimental clinical research.

All-Trans Retinoic Acid Prevents the Progression of Gastric Precancerous Lesions by Regulating Disordered Retinoic Acid Metabolism.

Retinoic acid (RA) is the most biologically active metabolite of vitamin A and is important for stomach physiological function. However, little is known about the metabolic status of RA in human gastric lesions. From 2015 to 2018, 1,392 local residents in Lujiang County were recruited into a cross-sectional survey program, which included a questionnaire interview and blood collection. We detected the mRNA and protein expression of RA metabolism-relevant factors in gastric tissues from 68 local patients with gastric lesions. The effects of all-trans retinoic acid (ATRA) supplementation were investigated in a gastric precancerous lesions (GPLs) rat model. In the cross-sectional survey, no significant differences in the level of RA precursor (P > 0.05) between the H. pylori seronegative and seropositive residents were observed. However, the mRNA and protein expression of RA synthesizing enzymes (RDH10 and ALDH1A1) were significantly decreased and catabolic enzyme (CYP26B1) was significantly increased in the patients (P < 0.05). Consistently, in the GPL rat model, we observed a similar disorder; however, ATRA supplementation significantly not only corrected the disorder by increasing Rdh10, Aldh1a1 and decreasing Cyp26b1, but also reduced claudin-18 (P < 0.05). Our study suggested that RA metabolism is disrupted in individuals with gastric lesions, while ATRA supplementation can prevent GPL from progressing to gastric cancer.

Impact of microecological agents on water environment restoration and microbial community structures of trench system in a Baiyangdian wetland ecosystem.

AIMS: To evaluate the effects of periodic application of effective microorganism (EM) bacteria and calcium peroxide on water environment restoration of a trench wetland ecosystem and its impact on microbiota in water and sediment. METHODS AND RESULTS: The EM bacteria and calcium peroxide were alternately sprayed into the trench water, and changes in the physicochemical indices of water and sediment, and in microbiota structure were subsequently investigated. Alternately spraying of calcium peroxide and EM bacteria improved the water quality, especially in terms of the dissolved oxygen and transparency of water body, and could further reduce the levels of total nitrogen, total phosphorus and ammonia nitrogen. At the same time, the microbiota structure of the water body was significantly changed by spraying with calcium peroxide and EM bacteria, and the relative abundances of Pseudanabaena, Legionellaceae, Planktothrix, Planctomyces, Phenylobacterium, Rhodobacter, Rhodoferax and Aquirestis were significantly increased. However, there was no significant effect on the physicochemical indexes and microbiota composition of the sediment. CONCLUSIONS: The water quality of the Baiyangdian trench could be significantly restored by regular sprinkling of calcium peroxide and EM bacteria. SIGNIFICANCE AND IMPACT OF STUDY: The results provide an effective technical method for the restoration of trench wetland water.

Epidemiological survey to determine the prevalence of cholecystolithiasis in Uyghur, Kazakh, and Han Ethnic Groups in the Xinjiang Uyghur Autonomous Region of China: cross-sectional studies.

BACKGROUND: This study was performed to understand the prevalence of and possible risk factors for cholecystolithiasis in Uyghur, Kazakh, Han, and other ethnic groups in the Xinjiang Uyghur autonomous region of China. METHODS: Subjects were enrolled using typical case sampling and multistage stratified random sampling. We collected epidemiological data regarding cholecystolithiasis using a standard questionnaire of risk factors for gallbladder disease in Xinjiang. The subjects completed the questionnaire and underwent an abdominal ultrasound examination of the liver and gallbladder. RESULTS: This study included 5454 Xinjiang residents aged ≥ 18 years. The prevalence of cholecystolithiasis was 15% (11.3% in men and 17.1% in women), and the sex difference was statistically significant (male-to-female odds ratio [OR] 1.867; p < 0.001). The cholecystolithiasis prevalence was also significantly different among the Han, Uyghur, Kazakh, and other ethnic groups (13.1%, 20.8%, 11.5%, and 16.8%, respectively; p < 0.001). The prevalence of cholecystolithiasis in northern Xinjiang was 13.5% and that in southern Xinjiang was 17.5%; this difference was also statistically significant (OR 1.599; p < 0.001). Across all ethnic groups, the cholecystolithiasis prevalence significantly increased with age (all p < 0.01) and body mass index (BMI) (all p < 0.01). A multivariate logistic regression analysis indicated that cholecystolithiasis prevalence was associated with sex, age, BMI, smoking, diabetes, fatty liver disease, and geographical differences between northern and southern Xinjiang. CONCLUSIONS: The prevalence of cholecystolithiasis was significantly higher in the Uyghur ethnic group than in the Han, Kazakh, and other ethnic groups; in women than in men; in southern Xinjiang than in northern Xinjiang; in patients with fatty liver disease; and increased with age and BMI. Our findings could provide a theoretical basis for the formulation of control measures for cholecystolithiasis.

Curcumin alleviates acute kidney injury in a dry-heat environment by reducing oxidative stress and inflammation in a rat model.

Curcumin exhibits anti-inflammatory and antioxidant activities. We investigated the protective effects of curcumin in a renal injury rat model under dry-heat conditions. We divided Sprague-Dawley rats into four groups: dry-heat 0- (normal temperature control group), 50-, 100-, and 150-minute groups. Each group was divided into five subgroups (n = 10): normal saline (NS), sodium carboxymethylcellulose (CMCNa), and curcumin pretreated low, medium, and high-dose (50, 100, and 200 mg/kg, respectively) groups. Compared to the normal temperature group, serum creatinine, blood urea nitrogen, urinary kidney injury molecule-1, and neutrophil gelatinase-associated load changes in lipoprotein (NGAL) levels were significantly increased in the dry-heat environment group (P < .05); inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) expression and malondialdehyde (MDA) and related inflammatory factor levels were increased in the kidney tissue. Superoxide dismutase (SOD) and catalase (CAT) levels were decreased. However, following all curcumin pretreatment, the serum levels of kidney injury indicators and NGAL were decreased in the urine compared to those in the NS and CMCNa groups (P < .05), whereas renal SOD and CAT activities were increased and MDA was decreased (P < .05). Renal tissues of the 150-minute group showed obvious pathological changes. Compared to the NS group, pathological changes in the renal tissues of the 100- and 200-mg/kg curcumin groups were significantly reduced. Furthermore, iNOS and COX-2 expression and inflammatory factor levels were decreased after curcumin treatment. Curcumin exerted renoprotective effects that were likely mediated by its antioxidant and anti-inflammatory effects in a dry-heat environment rat model.

Both TGF-ß1 and Smad4 regulate type I collagen expression in the muscle of grass carp, Ctenopharyngodon idella.

Type I collagen is proven to make an important contribution to fish muscle quality. Our previous study has shown the Smad4-dependent regulation of type I collagen expression in the muscle of crisp grass carp fed with faba bean. However, the regulatory roles of TGF-ß1 or TGF-ß1/Smad4 on type I collagen remain unclear in ordinary grass carp fed with normal diets or in other fish species. To clarify this point, the effect of TGF-ß1 and Smad4 over-expression and RNAi knockdown on type I collagen (COL1-α1 and COL1-α2) expression were tested in vitro (zebrafish ZF4 cells) and in vivo (grass carp) along with the TGF-ß1/Smad4 co-expression and co-knockdown. The mRNA levels of TGF-ß1, Smad4, and type I collagen were upregulated in the groups with over-expressed TGF-ß1 and Smad4 and downregulated in the groups of TGF-ß1 and Smad4 RNAi in comparison to controls in vitro (P < 0.05). Similarly, in the in vivo experiment, the mRNA abundance of TGF-ß1, Smad4, and type I collagen of over-expression group was higher than the controls at 36 h (P < 0.05). Co-injection of TGF-ß1/Smad4 over-expression and RNAi vectors generally showed the higher efficacy. This study revealed that TGF-ß1 and Smad4 genes regulated type I collagen expression in grass carp muscle and zebrafish. These findings will provide references for the collagen regulation of other freshwater fishes.

Comparative analysis of microbial community structure in the ponds with different aquaculture model and fish by high-throughput sequencing.

The pond has a complex microbial ecosystem, including microorganisms in water and sediment, which plays an important role in the health of fish and water quality. The microbial community structure in the ponds can be easily affected by many factors. However, not much is known about the role of different aquaculture model and fish on the microbial community structure in ponds. The purpose of the study was to investigate the microbial diversity and composition of the ponds with different aquaculture model and fish by high-throughput sequencing. A total of 3835072 valid sequences were achieved from 60 samples. Additionally, 2064 and 1917 core OTUs were observed in water and sediment samples, respectively. Our results suggested that sediment samples have a higher abundance and diversity of microbial community than water samples. In all the samples, the four most dominant phyla were Proteobacteria, Cyanobacteria, Actinomycetes and Bacteroides. At the genus level, hgcI_clade and CL500-29_marine_group were the dominant bacteria shared by the water samples and sediment samples. In addition, more bacteria related to eutrophication were found in the group of BF, BC and HSB, which suggested that these ponds may have been eutrophicated. In conclusion, the present study revealed the differences in the structure and diversity of microbial communities in ponds with different aquaculture model and fish. Furthermore, changes in typical bacteria of the ponds contribute to detect water quality and prevent water eutrophication.

Identification and expression analysis of miRNA in hybrid snakehead by deep sequencing approach and their targets prediction.

MicroRNAs (miRNAs) play important regulatory roles in numerous biological processes, but there is no report on miRNAs of hybrid snakehead. In this study, four independent small RNA libraries were constructed from the spleen, liver kidney and muscle of hybrid snakehead. These libraries were sequenced using deep sequencing technology, as result, a total of 1,067,172, 1,152,002, 1,653,941 and 970,866 clean reads from these four libraries were obtained. 252 known miRNAs and 63 putative novel miRNAs in these small RNA dataset were identified. The stem-loop RT-qPCR analysis indicated that eight known miRNAs and two novel miRNAs show different expression in eight different kinds of tissues. For better understanding the functions of miRNAs, 95,947 predicated target genes were analyzed by GO and their pathways, the results indicated that these targets of the identified miRNAs are involved in a broad range of physiological functions.

miR-214-5p targets KLF5 and suppresses proliferation of human hepatocellular carcinoma cells.

MicroRNAs (miRNAs) are small endogenous conserved RNAs regulating genes expression through base pairing with the 3'-untranslated region (3'-UTR) of target messenger RNAs. MiR-214-5p is a newly identified miRNA with its biological role largely unknown. In this study, we explored miR-214-5p expression status in 78 paired tumor and nontumor tissues obtained from patients with hepatocellular carcinoma (HCC) by RT-qPCR. The effects of miR-214-5p expression on HCC cell proliferation, cell cycle progression, and cell migration were measured by CCK-8 assay, flow cytometry, and wound-healing assay. A dual-luciferase activity assay was performed to identify whether KLF5 was a target of miR-214-5p. Kaplan-Meier curve and log-rank test were used to investigate the effects of miR-214-5p and KLF5 on overall survival and disease-free survival of patients with HCC. We found miR-214-5p expression was sharply reduced in HCC tissues and cell lines compared with the normal tissues and cell lines. Functional assay revealed that miR-214-5p overexpression could downregulate cell proliferation, cell migration, and arrested cell cycle at G0/G1 phase. Further, we validated Krüppel-like factor 5 (KLF5) as a direct target of miR-214-5p, and was upregulated in HCC and inversely correlated with the expression of miR-214-5p. Moreover, we found the low expression of miR-214-5p and high expression of KLF5 were correlated with tumor size, tumor stage, and poorer 5-year overall survival and disease-free survival of patients with HCC. In conclusion, our results suggested miR-214-5p functions as a tumor suppressor through targeting KLF5 in HCC. Also, miR-214-5p and KLF5 were identified as potential prognostic markers and might be therapeutic targets in HCC.

Lipid droplets participate in modulating innate immune genes in Ctenopharyngodon idella kidney cells.

Lipid droplets (LDs) are increasingly being recognized as important immune modulators in mammals, in additional to their function of lipid ester deposition. However, the role of LDs in fish immunity remains poorly understood. In this study, the function of LDs in the innate immune response of Ctenopharyngodon idella kidney (CIK) cells, which are the equivalent of myeloid cells in vertebrates, was investigated. LD number and TG content significantly increased in the CIK cells following exposure to lipopolysaccharide (LPS), peptidoglycan (PGN), and polyriboinosinic-polyribocytidylic acid (Poly [I: C]) for 24 h, accompanied by increases in the relative expression of several innate immune genes. However, fatty acid compositions of the triglycerides were not changed after treatment with these three pathogenic mimics. LPS, PGN, and Poly (I: C) did not alter the relative expressions of lipogenic (FAS, SCD, and DGAT) and lipid catabolic (PPARα, ATGL, and CPT-1) genes. However, these treatments did increase the mRNA levels of lipid transportation genes (FATP/CD36, ACSL1, and ACSL4), and also decreased the non-esterified fatty acid level in the medium. To further explore the role of LDs in the immune response, CIK cells were incubated with different concentrations (0, 100, 200, 300, 400, 500 µM) of exogenous lipid mix (LM; oleic acid [OA]:linoleic acid [LA]:linolenic acid [LNA] = 2:1:1), and were then transferred to a lipid-free medium and incubated for 24 h. LD size and number increased with the increase in lipid levels, and this was accompanied by increased expression of innate immune genes, including MyD88, IRF3, and IL-1ß, which were expressed at their highest levels in 300 µM exogenous lipid mix. Interestingly, after incubating with different fatty acids (LM, OA, LA, LNA, arachidonic acid [ARA], and docosahexaenoic acid [DHA]; 300 µM), ARA and DHA were more potent in inducing LD formation and innate immune gene expression in the CIK cells. Finally, atglistatin, an ATGL inhibitor, effectively attenuated the expression of most genes upregulated by ARA or DHA, suggesting that lipolysis may be involved in the regulation of immune genes at the transcriptional level. Overall, the findings of this study demonstrate that LDs are functional organelles that could act as modulators in the innate immune response of CIK cells. Additionally, long-chain polyunsaturated fatty acid enriched LDs play a unique role in regulating this process.