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In developing a bisulfite-free methylation assay with the use of restriction enzymes, self-ligation of next generation sequencing adapters (NGS-adapter) is a technological bottleneck to be overcome. In the experiments of this study, a variety of strategies designed to limit or abolish adapter’s self-ligation has been tested. Experimental data have showed that the three strategies tested can either substantially decrease or completely abolish the self-ligation of NGS-adapters. Minimization or elimination of NGS-adapter’s self-ligation is of importance in increasing the sensitivity, efficiency, and reproducibility of enzyme-mediated methylation assay. The strategies reported in the present study may find applications in some other nanotechnologies. In combination with nanotechnologies, either drop-digit PCR or microarray-based sequencing, the methylation-dependent endonuclease mediated methylation assays will facilitate applications of methylation analysis in both fundamental research and clinical epigenetic studies, particularly in early diagnosis of cancer.

In our previous study, serum soluble T-cell immunoglobulin and mucin structure-3 (sTim-3) and galactosin-9 (sGal-9) were found to be associated with renal function after kidney transplantation. However, it is unclear whether these two indicators can predict adverse outcomes after transplantation.Ninety-one recipients of kidney transplantation were enrolled and divided into a stable group and an adverse outcome group (consisting of biopsy-proven rejection, graft loss, death and clinically diagnosed rejection). The expression levels of sTim-3 and sGal-9 before (pre-Tim-3 and pre-Gal-9) and one month after transplantation (post-Tim-3 and post-Gal-9) were measured by ELISA.The level of pre-Tim-3 was significantly higher in the stable group than in the adverse outcome group [median (range), 2275 (840-4236) pg/mL vs. 1589 (353-3094) pg/mL, P = 0.002]. The level of post-Gal-9 was significantly lower in the stable group than in the adverse outcome group [median (range), 4869 (1418-13080) pg/mL vs. 6852: (4128-10760) pg/mL, P = 0.003]. The areas under the curve (AUCs) for pre-Tim-3 and post-Gal-9 were 0.737 (P = 0.002) and 0.751 (P = 0.003), respectively, better than AUC of post-eGFR (0.633) (P = 0.071), according to the receiver operating characteristic (ROC) curve. Through Cox regression analysis, including pre-Tim-3, post-Gal-9, post-eGFR, sex, age, BMI of recipients and donors, pre-Tim-3 and post-Gal-9 were independent risk factors for adverse outcomes after kidney transplantation (P = 0.016, P = 0.033, respectively).Serum sTim-3 and sGal-9 can predict adverse outcomes within two years after kidney transplantation.

Turpiniae Folium, the dried leaves of Turpinia arguta Seem., is a kind of historic traditional Chinese medicine. Here, based on our previous study, we extracted the Turpiniae Folium polysaccharides (TFP) and isolated three polysaccharide fractions from TFP. Then, TFP and one of the major polysaccharide fractions (TFP-1a) were identified through HPLC, HPGPC, and ATR-FTIR. Furthermore, the evaluations of their antioxidative, anti-inflammatory activities and inhibitory effect on angiotensin II-induced vascular smooth muscle cells (VSCMs) proliferation in vitro were conducted. Both TFP and TFP-1a showed strong hydroxyl radical scavenging, DPPH radical scavenging, and Fe

Glacial debris flows (GDFs) often occur in alpine regions that are subject to rapid climate change, and pose a serious threat to road systems. However, the ways that climate change impacts GDF risks along road systems remain poorly understood. Aierkuran Gully, located in eastern Pamir along Karakoram Highway (KKH), is a hotspot for GDF activity and climate change, and was thus selected to investigate the GDF risk to road systems under climate change conditions. RegCM4.6 climate data for northwestern China were selected as climate projections during baseline (2011–2020) and future periods (2031–2040) under the Representative Concentration Pathway (RCP) 8.5. To reflect the coupling effect of rainfall and melt water that triggers GDF, a glacial hydrological model DETIM that considers both factors was applied to calculate the peak debris flow discharge. A FLO-2D model was calibrated based on high-quality data collected from a detailed field investigation and historical debris flow event. The FLO-2D model was used to simulate the debris flow depth and velocity during baseline and future periods under RCP8.5. The debris flow hazard was analyzed by integrating the maximum flow depth and momentum. Road structure vulnerability was further determined based on the economic value and susceptibility of hazard-affected objects. The GDF risk along KKH was assessed based on the GDF hazard and vulnerability analysis. Our results show that climate change would lead to amplified peak debris flow discharge, trigger higher-magnitude GDF, and induce more severe damage and threats to the road system. Compared with the baseline period, the debris flow damage risk for culverts and bridges would increase and the areas that inundate the road and pavement would expand. Our findings provide valuable insights for the development of mitigation strategies to adapt road systems to climate change, especially in alpine regions with highly active GDFs.

Lonicerae Japonicae Flos and Lonicerae Flos, as traditional Chinese medicinal and edible food, are widely used in medicine, food, health products, and other industries. However, there is no comprehensive study on the differences of flavor compounds in Lonicerae Japonicae Flos and Lonicerae Flos. This study applied headspace gas chromatography-ion mobility spectrometry(HS-GC-IMS) to analyze the differences of flavor compounds in Lonicerae Japonicae Flos and Lonicerae Flos. The differential biomarkers were confirmed by multivariate statistical analysis. The results showed that there were significant differences in the forty-seven flavor compounds in Lonicerae Japonicae Flos and Lonicerae Flos. The differential markers were ethyl acetate, propyl alcohol, 1-octanol, 1-hexanol, hexanal, and(Z)-2-hexen-1-ol. Pathway enrichment analysis showed that the above markers were involved in the biosynthesis of major secondary metabolism, sulfate metabolism pathways, and formation of other flavor compounds. This study provides important references for the evaluation of flavor compounds of Lonicerae Japonicae Flos and Lonicerae Flos and the development of medicinal and edible products.

Lonicera japonica Thunb. (LJ) and Lonicera macranthoides Hand. -Mazz. (LM) have been widely used in Chinese medicine for thousands of years. Although the morphological characteristics of LJ and LM are quite similar, there are significant distinctions of medicinal ingredients (mainly the secondary metabolites) and clinical indications between them. However, the in-depth differences of primary metabolites have not thoroughly been studied yet. Therefore, gas chromatography-mass spectrometry- (GC-MS-) based metabolomics method combined with chemometric methods were performed to analyze the distinction in this study. The results showed that LJ and LM were obviously classified into two groups. 10 metabolites were obtained as biomarkers on account of their p values, pcorr values, and differing variable importance in projection (VIP) values. Metabolic pathway analysis showed that the galactose metabolism and starch and sucrose metabolism gathered as potential pathways caused these extraordinary differences of primary metabolites between LJ and LM. Further, we found that the differences of main medicinal ingredients between LJ and LM could be interpreted from these metabolites according to the analysis of mainly related pathways. The metabolites involved in the starch and sucrose metabolism presented upregulated in LJ, while almost all metabolites in the galactose metabolism, the TCA cycle, and the phenolic acid part of phenylpropanoid metabolism were downregulated in LJ. Therefore, the energy stored in the starch and sucrose metabolism may be saved to produce flavonoid, which could be the reason that the level of flavonoid of phenylpropanoid metabolism is higher in LJ compared to LM. Consequently, this study presented an effective tool for quality evaluation of LJ and LM and laid a foundation for further studies of the metabolic mechanisms and high-quality manufacturing of them.

To identify the differences in the composition of gut microbiota of patients with post-stroke cognitive impairment (PSCI) in comparison with the normal cognition healthy controls (HC), and to study the potential association between gut microbiota and cognition function.A total of 24 patients were recruited for the PSCI group, which was matched with 23 healthy subjects with no history of cardiovascular disease recruited over the same period for the control group. Fecal samples were collected for both groups, and Mini-Mental State Examination (MMSE) and Montreal Cognitive Assessment (MoCA) were used to evaluate cognitive functions. The abundance, diversity and group difference of gut bacterial communities were determined with 16S rRNA gene sequencing, and the correlations between differences in bacterial species of the gut microbiota and cognitive function scores were examined with redundancy analysis (RDA)/canonical correspondence analysis (CCA).There was no significant difference in the general data or the alpha diversity of gut microbiota between the two groups (In this study, we observed compositional differences between the gut microbiota of PSCI patients and those of HC, and revealed that the differences were correlated, to some degree, to the cognitive functions, which will provide new perspectives for the clinical diagnosis and treatment of PSCI.

A series of novel oleanolic acid (OA)-linked disulfide, thioether, or selenium ether derivatives was synthesized, and their antiproliferative activity was evaluated against human liver cancer (BEL-7402 and HepG-2), colon cancer (HCT116), and normal liver (L02) cell lines using methyl thiazolyl tetrazolium assay (MTT). Preliminary bioassay results revealed that OA derivatives modified at the C3-OH position, i. e., compound a4 containing sulfide ether, exhibited the best antiproliferative activity against BEL-7402 cells, with an IC

Objective Hedyotis corymbosa (Shui Xian Cao, 水线草, HC) is the main antitumor medicine in China and other southeast Asian countries. A comprehensive quality control system of HC was established in this study, as the current system was ineffective.Methods Microscopy was used to observe the characteristics of the powder and cross-sectional tissue. Thin layer chromatography (TLC) was employed to separate HC from Hedyotis diffusa (Bai Hua She She Cao, 白花蛇舌草, HD). Simultaneously, high-performance liquid chromatography (HPLC) was used to establish the characteristic spectrum of HC and determine the concentrations of hedyotiscone A. Regarding the issue of systematicness, concentrations of water, total ash, acid-insoluble ash, heavy metals (including lead, arsenic, mercury, cadmium and copper) and pesticide residue were measured by the methods described in the 2015 edition of Chinese Pharmacopoeia.Results Calcium oxalate needle crystal bundles and oil drops were observed in the cross-sectional tissue. Three characteristic blue fluorescent spots were observed in the HC samples by TLC. There were five distinct characteristic peaks in HC but not in HD, and hedyotiscone A was the sharpest characteristic peak. The generated linear equation of hedyotiscone A was Y = 3.15 × 108X + 312.04 (r2 = 1.000) between 0.00005270 and 0.002108 mg/mL, and the average recovery was 100.42%. Precision [relative standard deviation (RSD) 0.92%], repeatability (RSD 0.14%), and recovery (RSD 0.85%) met the requirements for quantitative analysis. Hedyotiscone A concentrations ranged from 0.00017%∼0.00113% across different habitats. Concentrations of water, ash and acid-insoluble ash were 8.80%∼11.2%, 7.00%∼15.3% and 0.440%∼7.70%, respectively. Concentrations of Pb, As, Hg, Cd and Cu were 1.60∼4.60, 0.650∼1.60, 0.0300∼0.100, 0.240∼0.690 and 10.1∼16.6 µg/g, respectively. Pesticide residues were less than 0.00100 ppm.Conclusion The quality control system can effectively distinguish HC from HD, so as to comprehensively control the quality of HC, to provide references for the clinical use of HC. Keywords: Hedyotis corymbosa, Hedyotis diffusa, Hedyotiscone A, TLC, HPLC, Quality standard

Fluoride is considered as one of the most ubiquitous environmental pollutants. Numerous studies have linked reactive oxygen species (ROS)-dependent oxidative damage with fluoride intoxication, which could be prevented by antioxidants. However, the metabolomic changes induced by ROS disruptions in fluoride intoxication are yet unknown. The present study aimed to provide novel mechanistic insights into the fluoride-induced oxidative damage and to investigate the potential protective effects of ethanolic extract of Prunella vulgaris (natural antioxidant, PV) against fluoride-induced oxidative damage. The serum biochemical indicators related to fluoride-induced oxidative damage, such as lipid peroxidation parameter, inflammation and marker enzymes in the liver increased significantly in the fluoride-treated group, while antioxidant enzymes were decreased. However, PV treatment restored the level of these biochemical indicators, indicating satisfactory antioxidant, anti-inflammatory, and hepatoprotective potential of PV. The metabolomics analysis in the serum was performed by liquid chromatography-mass spectroscopy, whereas the fluoride treatment caused severe metabolic disorders in rats, which could be improved by PV. The differential metabolites screened by multivariate analysis after fluoride and PV treatment, were organic acids, fatty acids, and lipids. These differential metabolites represented disorders of glyoxylate and dicarboxylate metabolism and the citrate cycle (TCA) according to metabolic pathway analysis in fluoride treatment rats. Interestingly, the result of metabolic pathway analysis of post-treatment with PV was consistent with that of fluoride treatment, indicating that the energy metabolism plays a major role in the progress of fluoride-induced oxidative damage, as well as the therapeutic effect of PV. These findings provided a theoretical basis for understanding the mechanism underlying metabolic disorders of fluoride toxicity and the effect of PV.

Prunella vulgaris(PV) is an edible and traditional medicinal herb which has a wide range application in fighting inflammation and oxidative stress, and protecting liver. Now it has been used to treat various types of liver diseases and has significant clinical efficacy. This study aims to investigate the effects of PV on ethanol-induced oxidative stress injury in rats and its metabolic mechanism. The rats were divided into control group, model group, PV group, and VC group. The liver protection of PV was identified by measuring pharmacological indexes such as antioxidant and anti-inflammatory activity. The metabolic mechanism of long-term ethanol exposure and the metabolic regulation mechanism of PV treatment were studied by LS-MS metabonomics. The pharmacological investigation indicated that ethanol could significantly decrease the contents of SOD, GSH-Px, CAT and other antioxidant enzymes in liver and increase the content of MDA. At the same time, PV could significantly reduce the contents of inflammatory factors(TNF-α, IL-6 and IL-1β) and liver function markers(ALT, AST, ALP) in serum. What's more, long-term ethanol exposure could significantly cause liver injury, while PV could protect liver. Metabolomics based on multiple statistical analyses showed that long-term ethanol exposure could cause significant metabolic disorder, and fatty acids, phospholipids, carnitines and sterols were the main biomarkers. Meanwhile, pathway analysis and enrichment analysis showed that the β oxidation of branched fatty acids was the main influencing pathway. Also, PV could improve metabolic disorder of liver injury induced by ethanol, and amino acids, fatty acids, and phospholi-pids were the main biomarkers in PV treatment. Metabolic pathway analysis showed that PV mainly regulated metabolic disorder of ethanol-induced liver injury through phenylalanine, tyrosine and tryptophan biosynthetic pathways. This study could provide a new perspective on the hepatoprotective effect of natural medicines, such as PV.

Metabolomics is applied to explore the curative effect of complex systems, such as Chinese medicine. Intrauterine adhesion (IUA) harms the reproductive system and affects fertility, and hence is a significant public health concern. Prunella vulgaris oil (PVO) protects the reproductive system and exerts anti-inflammatory effects, but its effect on IUA and the underlying mechanism is unclear. In this study, we established a serum metabolomics method based on GC-TOF-MS to evaluate the mechanism of PVO in the IUA rat model established by mechanical injury and infection. Animal experiments showed that PVO improves the inflammatory response in the uterus of IUA model rats and reduces the content of inflammatory factors to improve the microenvironment of the reproductive system. It also regulates the expression of TGF-β1 and Smad-related mRNA and protein to inhibit fibrosis. Metabolomics indicated a significant abnormality in serum metabolism in IUA rats, and a total of 51 differential markers were screened and identified. After PVO treatment, these metabolic abnormalities improved significantly. The metabolic pathway analysis revealed that PVO affects glyoxylate and dicarboxylate metabolism, and β-alanine metabolism pathways. This study showed that PVO significantly improves inflammation and fibrosis in IUA rats combined with the pharmacological results. The primary mechanism is related to regulating the metabolism of amino acids and their derivatives to balance the associated disorders and control energy metabolism.

Hyperlipidemia, a typical metabolic disorder syndrome, can cause various cardiovascular diseases. The polysaccharides were found to have enormous potential in the therapy of hyperlipidemia. This study was aimed at evaluating the ameliorative effects of polysaccharide from Turpiniae folium (TFP) in rats with hyperlipidemia. A serum metabolomic method based on gas chromatography-mass spectrometry (GC-MS) was used to explore the detailed mechanism of TFP in rats with hyperlipidemia. The oxidative stress indicators, biochemical indexes, and inflammatory factors in serum and histopathological changes in the liver were also evaluated after 10-week oral administration of TFP in rats with high-fat diet-induced hyperlipidemia. TFP significantly relieved oxidative stress, inflammation, and liver histopathology and reduced blood lipid levels. Multivariate statistical approaches such as principal component analysis and orthogonal projection to latent structure square-discriminant analysis revealed clear separations of metabolic profiles among the control, HFD, and HFD+TFP groups, indicating a moderating effect of TFP on the metabolic disorders in rats with hyperlipidemia. Seven metabolites in serum, involved in glycine, serine, and threonine metabolism and aminoacyl-tRNA biosynthesis, were selected as potential biomarkers in rats with hyperlipidemia and regulated by TFP administration. It was concluded that TFP had remarkable potential for treating hyperlipidemia. These findings provided evidence for further understanding of the mechanism of action of TFP on hyperlipidemia.

To evaluate the predictive performance of anthropometric indices for metabolic syndrome (MetS) among Chinese adolescents with different nutritional status.We recruited 9,513 adolescents aged 10-18 years from seven provinces in China during September 2014. Anthropometric indices and blood pressure were measured at recruitment, and blood samples were collected for determining fasting plasma glucose and lipid profile. Receiver operating characteristic (ROC) analyses were used to assess the predictive performance of anthropometric indices, including body mass index (BMI) percentile, waist circumference percentile, waist-height ratio, and waist-hip ratio.Overall, the four anthropometric indices showed good accuracy for predicting MetS with areas under ROC curves (AUCs) ranging from 0.86 to 0.94; similar AUCs ranging from 0.73 to 0.99 were observed for participants with normal weight. The performance of all four indices was poor in overweight and obese participants, with AUCs ranging from 0.66 to 0.77 and from 0.60 to 0.67, respectively. Waist circumference showed relatively better performance in all the subgroup analyses.We suggest using anthropometric indices with the cutoff values presented here for predicting MetS in the overall and normal-weight adolescent population, but not in the overweight and obese adolescent population where more specific screening tests are required.

Prunella vulgaris L., better known as 'self-heal', has been extensively used in the traditional system of medicines. To reveal the regulatory mechanism of its development, TMT-based quantitative proteome analysis was performed in the Prunella vulgaris L. spica before and during ripening (Group A and Group B, respectively). This analysis resulted in the identification of 7655 proteins, of which 1910 showed differential abundance between the two groups. Pronounced changes in the proteomic profile included the following: 1) Stress-responsive proteins involved in protecting cells and promoting fruit ripening and seed development were highly abundant during ripening. 2) The degradation of chlorophyll, inhibition of chlorophyll biosynthesis and increased abundance of transketolase occurred simultaneously in the spica of Prunella vulgaris L., resulting in the spica changing color from green to brownish red. 3) The abundance of protein species related to phenylpropanoid biosynthesis mainly increased during ripening, while flavonoid and terpenoid backbone biosynthesis mostly occurred before ripening. SIGNIFICANCE: This study establishes a link between protein profiles and mature phenotypes, which will help to improve our understanding of the molecular mechanisms involved in the maturation of Prunella vulgaris L. at the proteome level and reveal the scientific connotation for the best time to harvest Prunella vulgaris L. This work provides a scientific basis for the production of high-quality medicinal Prunella vulgaris L., as well as a typical demonstration of molecular research used for the harvest period of traditional Chinese medicine. BIOLOGICAL SIGNIFICANCE: This work provided a comprehensive overview on the functional protein profile changes of Prunella vulgaris L. spica at different growing stages, as well as the scientific rationale of Prunella vulgaris L. harvested in summer after brownish red, thus laid an intriguing stepping stone for elucidating the molecular mechanisms of quality development.

Hyperlipidemia, a typical metabolic disorder syndrome, can cause various cardiovascular diseases. The polysaccharides were found to have enormous potential in the therapy of hyperlipidemia. This study was aimed at evaluating the ameliorative effects of polysaccharide from Turpiniae folium (TFP) in rats with hyperlipidemia. A serum metabolomic method based on gas chromatography-mass spectrometry (GC-MS) was used to explore the detailed mechanism of TFP in rats with hyperlipidemia. The oxidative stress indicators, biochemical indexes, and inflammatory factors in serum and histopathological changes in the liver were also evaluated after 10-week oral administration of TFP in rats with high-fat diet-induced hyperlipidemia. TFP significantly relieved oxidative stress, inflammation, and liver histopathology and reduced blood lipid levels. Multivariate statistical approaches such as principal component analysis and orthogonal projection to latent structure square-discriminant analysis revealed clear separations of metabolic profiles among the control, HFD, and HFD+TFP groups, indicating a moderating effect of TFP on the metabolic disorders in rats with hyperlipidemia. Seven metabolites in serum, involved in glycine, serine, and threonine metabolism and aminoacyl-tRNA biosynthesis, were selected as potential biomarkers in rats with hyperlipidemia and regulated by TFP administration. It was concluded that TFP had remarkable potential for treating hyperlipidemia. These findings provided evidence for further understanding of the mechanism of action of TFP on hyperlipidemia.

Background The flowers and dried fruit spikes of Prunella vulgaris L. (P. vulgaris L.) have been widely used in traditional Chinese medicine and food. P. vulgaris L. is regarded as a good option for treating uterine myoma (UM). However, scientific evidence of anti-UM activity of the extract of P. vulgaris L. (PVE) is lacking. The present study aimed to characterize the chemical composition of PVE and evaluate the pharmacodynamics and mechanism of PVE against UM. Methods The chemical composition of PVE was analyzed by GC-MS. MTT was used to screen and evaluate cell proliferation and toxicity. Double fluorescence flow cytometry method were used to determine the apoptosis and cell cycle progression of UM cells under PVE treatment. The anti-UM activity of PVE was investigated by using a specific-pathogen-free (SPF) rat model of UM. TUNEL staining was used to detect the apoptosis of UM cells. The concentrations of estrogen and progesterone in the serum of SPF rats were detected by ELISA. The expression levels of PCNA, estrogen receptor alpha, estrogen receptor beta, progesterone receptor, survivin, caspase-3, Bax and Bcl-2 in the uterus of SPF rats was detected by immunohistochemistry (IHC). Results The extraction rate of PVE was 8.1%. The main components were squalene (28.3%), linoleic acid (9.96%), linolenic acid (9.95%), stearic acid (6.26%) and oleic acid (5.51%). In vitro, PVE had significant anti-human UM cell activity, exhibited no drug toxicity, promoted the apoptosis of human UM cells, and inhibited the transition of UM cells from the G0/G1 stage into the G2 stage, in which DNA replication occurs. In vivo, PVE had significant anti-UM activity. PVE decreased the concentrations of estrogen and progesterone and downregulated the expression levels of the estrogen and progesterone receptors through the estrogen signaling pathway. PVE also promoted the apoptosis of UM cells by downregulating the expression levels of the survivin and Bcl-2 proteins and upregulating the expression levels of caspase-3 and Bax through the mitochondria-mediated apoptotic pathway. Conclusion PVE has marked anti-UM activity. PVE can be used as an ideal candidate drug to treat UM.

Pelvic inflammatory disease( PID) rat model was induced by the mixture of Escherichia coli,Staphylococcus aureus,and Streptococcus hemolytic-β． Gas chromatography-mass spectrometry( GC-MS) based metabolic profiling method was combined with multivariate statistical analysis,such as PCA,PLS-DA and OPLS-DA to analyze endogenous small molecule metabolites in serum of rats after treatment of Fuke Qianjin Capsules. The results showed that Fuke Qianjin Capsules could significantly improve the inflammatory pathological characteristics and tissue damages in model rats. Based on the principle of VIP1 and P0. 05,a total of 6 different metabolic biomarkers were identified,including L-valine,L-isoleucine,L-threonine,butanedioic acid,serine and D-glucose,respectively.The contents of these six different metabolites were significantly reversed after administration. Further analysis of the metabolite pathways through KEGG database showed that Fuke Qianjin Capsules achieved the effect possibly through glycine,serine and threonine metabolism,aminoacyl-tRNA biosynthesis and valine,leucine and isoleucine biosynthesis. Therefore,this study came to the conclusion that Fuke Qianjin Capsules can be used in the treatment of mixed bacteria induced pelvic inflammatory disease possibly by regulating amino acid and its derivative metabolism.

Background T cell immunoglobulin mucin-3 (Tim-3) has been reported to participate in the regulation of immune response and the induction of allograft tolerance. However, the association between Tim-3 and renal allograft dysfunction is unclear. We studied the expression of cellular and soluble Tim-3 (sTim-3), soluble galectin-9 (sGal-9) and carcinoembryonic antigen-related cell adhesion molecule-1 (sCEACAM-1) in kidney transplantation recipients (KTRs) to explore their roles in allograft dysfunction. Methods 96 KTRs (53 with stable graft and 43 with graft dysfunction) and 30 healthy controls (HC) were enrolled. Among the KTRs, 55 used Tacrolimus (TAC) and 41 used Sirolimus (SRL). In the dysfunction group, 29 recipients have undergone graft biopsy and 14 were classified as biopsy-proven rejection (BPR). Cellular Tim-3 was determined by flow cytometry. sTim-3 was determined by ELISA. sGal-9 and sCEACAM-1 were determined by Bio-Plex® suspension array system. Results KTRs with renal dysfunction showed significantly higher levels of sTim-3 and sGal-9 but similar levels of cellular Tim-3 and sCEACAM-1 compared with stable recipients. Correlation analysis revealed that estimated glomerular filtration rate (eGFR) was negatively associated with sTim-3 and sGal-9. Both BPR and non-BPR groups showed comparable levels of Tim-3, Gal-9 and CEACAM-1. Moreover, SRL group showed significantly higher levels of sCEACAM-1 than TAC and HC groups. Conclusions sTim-3 and sGal-9 were promising biomarkers for allograft dysfunction, but unable to differentiate allograft rejection from other causes of renal dysfunction in KTRs. Moreover, long-term administration of sirolimus would up-regulate sCEACAM-1 level, while exert similar regulatory effects on Tim-3 and Gal-9 compared to tacrolimus.

Objectives In alcoholic liver disease, alcohol and lipopolysaccharide (LPS) are major stimulation factors of hepatic lipogenesis. Our objective was to determine the protective mechanism of acanthoic acid (AA) in EtOH- and LPS-induced hepatic lipogenesis. Methods HSC-T6 cells were treated with ethanol (200 mm) plus LPS (1 μg/ml) for 1 h, followed by AA (10 or 20 μm) for another 6 h. C57BL/6 mice were pretreated with of AA (20 and 40 mg/kg) or equal volume of saline and then exposed to three doses of ethanol (5 g/kg body weight) within 24 h. The mice were sacrificed at 6 h after the last ethanol dosing. Key findings Acanthoic acid significantly decreased the expressions of α-SMA, collagen-I, SREBP-1, and lipin1/2 induced, also decreased fat droplets caused by EtOH/LPS. AA treatment decreased the protein expressions of TLR4, CD14, IRAK4, TRAF3, p-TAK1 and NF-κB increased by EtOH/LPS on HSC cells. Results in vivo were consistent with results in vitro. Conclusions Our data demonstrated that AA might modulate hepatic fibrosis and lipid deposition in HSC-T6 cell stimulated with ethanol combined with LPS by decreasing lipin1/2 via TLR4 and IRAK4 signalling pathways, and AA might be considered as a potential therapeutic candidate for alcoholic liver disease.

BACKGROUND We investigated whether a low fixed Tac starting dose regimen could lead to a better achievement of Tac target concentrations, as well as an effective immunosuppressive treatment, in Chinese kidney transplant recipients (KTRs). MATERIAL AND METHODS We collected whole-blood and serum samples from 189 KTRs and the Tac starting dose was 2, 2.5, or 3 mg/day. Information on Tac C0, dose, body weight, body mass index (BMI), Scr, eGFR, and CYP3A5 genotypes were collected from a routine therapeutic drug monitoring database. The correlation between Tac C0 and body weight (or BMI) was investigated by calculating the goodness of fit. Multivariable logistic regression was performed to estimate the independent associated factors. RESULTS The patients with 3 mg per day of Tac had higher C0 at day 7 compared to those with 2 or 2.5 mg. For patients receiving the same Tac starting dose, no significant difference was found in Tac C0 at day 7 among different body weight or BMI groups. There was no significant difference in Scr or eGFR at 1 year after transplant, nor was there a significant difference in the rates of DGF or AR at post-transplant day 30 among different Tac starting dose groups or among the 3 Tac C0 range groups. CYP3A5 genotype and Tac initial dose were independently associated with Tac C0. CONCLUSIONS CYP3A5 genotype and Tac initial dose were independently associated with Tac C0 in renal transplant recipients. Our results suggest that a low Tac target C0 range (5-8 ng/ml) with a low fixed starting dose (3 mg/day) would be safe and effective among Chinese KTRs.

The aim of this study was to investigate the effects of acanthoic acid (AA) on the regulation of inflammatory response, lipid accumulation, and fibrosis via AMPK- IRAK4 signaling against chronic alcohol consumption in mice. Ethanol-induced liver injury was induced in male mice by Lieber-DeCarli diet for 28d. And mice in AA groups were gavaged with AA (20 or 40mg/kg) for 28d. AA treatment significantly decreased serum AST and TG, hepatic TG levels, serum ethanol and LPS levels compared with chronic ethanol administration. AA ameliorated histological changes, lipid droplets, hepatic fibrosis, and inflammation induced by ethanol. AA significantly increased the expressions of p-LKB1, p-AMPK, and SIRT1 caused by chronic ethanol administration, and attenuated the increasing protein expressions of IRAK1 and IRAK4.siRNA against AMPKα1 blocked AMPKα1 and increased IRAK4 protein expressions, compared with control-siRNA-transfected group, while AA treatment significantly decreased IRAK4 expressions compared with AMPKα1-siRNA-transfected group. AMPK-siRNA also blocked the decreased effect of AA on inflammatory factors. AA decreased over-expression of IRAK4 and inflammation under ethanol plus LPS challenge. AA recruited LKB1-AMPK phosphorylation and activated SIRT1 to regulate alcoholic liver injury, especially, inhibited IRAK1/4 signaling pathway to regulate lipid metabolism, hepatic fibrosis and inflammation caused by alcohol consumption.

Two new diterpenoids, neoorthosiphonones B and C (1 and 2), and one known diterpenoid, were isolated from the aerial parts of Clerodendranthus spicatus. Their structures including absolute configurations were determined by comprehensive spectroscopic analyses and X-ray crystallographic methods. No compound was found to inhibit fibronectin production at the concentration of 20 μM. Electronic supplementary material The online version of this article (doi:10.1007/s13659-017-0128-8) contains supplementary material, which is available to authorized users.