Gastric cancer (GC) is one of the most common malignancies worldwide. The expression of CD147 protein is associated with GC. However, the clinical role of CD147 in GC has not been investigated extensively. Hence, we focused on studying the association between the expression of CD147 and clinicopathological features of GC patients in this study. Firstly, sixteen publications (1752 cases and 391 controls) and one from our own original research (143 cases) were included in the meta-analysis to obtain a more precise estimation of the diagnostic value of CD147. The results showed that expression rate of CD147 in the GC group is higher than that in control group. Moreover, gender, TNM stage, lymph node metastasis, and depth of invasion are all associated with CD147. Further, sections of gastric tissue from 143 cases underwent immunohistochemical staining for evaluation of CD147 protein expression. Our retrospective analysis demonstrated CD147 protein expression was significantly associated with clinical N stage, and tumor stage. Meanwhile, it can also serve as an independent prognosis biomarker. In conclusion, our results support the role of CD147 as a good indicator of diagnosis and prognosis. Gastric cancer (GC) is one of the most common malignancies in the world and the second leading cause of global cancer death . Based oExtracellular matrix metalloproteinase inducer, also named as cluster of differentiation 147 (CD147) or basigin, is a widely distributed cell surface glycoprotein that is involved in numerous physiological and pathological functions, especially in tumor invasion and metastasis . RiethdoIn fact, meta-analysis can play a key role in generating new hypotheses . In thisSixteen publications were included in our meta-analysis. Ten studies were included for overall analysis and other six studies were included for subgroup analysis. The screening process is presented in detail in Figure P = 0.000, I2 = 86.2%, Figure P = 0.000, I2 = 89.3%, Figure A total of 10 studies , 10–17 . The stability of the study was detected by sensitivity analysis, by excluding one individual study in sequence. Our analysis displayed that result was not changed.P = 0.011), and tumor stage .We calculated the expression levels of CD147 protein in 143 paraffin-embedded GC samples using immunohistochemical staining. The power analysis indicates our sample size is enough (Power = 0.9997). As shown in Figure P = 0.0001, Figure P = 0.001, Figure In order to investigate the association between CD147 expression and clinical outcome, we divided the patients into CD147-high and CD147-low groups, and survival analyses were performed using the Kaplan-Meier method. In summary, the patients with higher CD147 expression had significant lower OS (P < 0.0001) than those with lower CD147 expression Figure . MeanwhiP = 8.95E-08). In multivariate survival analysis, T, N, and M collapsing into stage as a whole, together with age and gender, were included in the Cox proportional hazard models as adjusted variables. Significantly, overexpression of CD147 is negatively correlated with OS , which indicated that CD147 can act as an independent predict factor for GC patients (Table P = 1.66E-05) which has been universally acknowledged   and one original study (143 cases) from our own research, to assess the association between CD147 and the clinicopathological features of GC. Our results indicate that expression rate of CD147 in the GC group is higher than that in control group. Besides, gender, TNM stage, lymph node metastasis, and depth of invasion have a relationship with the expression state of CD147. Further, our retrospective analysis of 143 cases demonstrated CD147 protein expression can be an independent prognosis biomarker for GC with a strong statistical power.In recent years, although surgery, chemoradiotherapy, and targeted therapy make great progress to prolong survival of GC patients, GC is still the third leading cause of cancer-related death worldwide mainly due to tumor local recurrence and distant metastasis . With thCD147 is a 57-kilodalton (kDa) transmembrane glycoprotein which belongs to the immunoglobulin (IgG) superfamily . MoreoveBased on our meta-analysis, results showed that the expression rate of CD147 in the GC group was higher than that in control group. Furthermore, CD147 expressions in GC tissues were significantly higher than normal mucosa tissues. When exploring the diagnostic value of CD147 in GC by immunohistochemistry, we found that CD147 expression level was higher in GC tissues than in adjacent non-tumor tissues. Similar with our study, Xu et al.  and Wu ePrevious studies have identified gender differences that may contribute to the development of GC , 36. It Invading basement membrane (BM) barriers is essential steps in the pathology of GC . Depth oThe TNM Classification of Malignant Tumors (TNM) is a cancer staging notation system that gives codes to describe the stage of a person's cancer. The higher stages represent higher progression of cancer. Previously, Wu et al.  indicateAfter analyzing the cases with clinic information in our retrospective data, we found that the OS of the GC group with low CD147 expression had a longer survival time than that of high CD147 expression group. The univariate and multivariate analyses also demonstrated that CD147 overexpression was significantly associated with poor OS. Notably, the patients with lower CD147 expression showed a longer survival time compared with high CD147 expression patients in low stage group and high stage group. This may indicate that the level of CD147 expression can be used as a molecular biomarker for the prognosis of GC. Since good prognostic prediction is necessary to ascertain the risk and the effectiveness of treatments such as surgery and chemoradiotherapy, CD147 may be a competent prognostic marker to predict treatment response.However, several limitations about our research should be addressed. First, there are high heterogeneity in GC group vs. control group in the meta-analysis. After we excluded Pinheiro et al.'s study , I squarAbove all, CD147 upregulated in GC tissues compared with noncancerous tissues. Moreover, gender, TNM stage, lymph node metastasis, and depth of invasion are all associated with CD147. Our retrospective analysis demonstrated CD147 protein expression was significantly associated with clinical N stage, and tumor stage. Meanwhile, it can also serve as an independent prognosis biomarker. In conclusion, our results support the role of CD147 as a good indicator of diagnosis and prognosis.Eligible publications were retrieved by searching PubMed, Embase, and Chinese National Knowledge Infrastructure (CNKI) databases up to April 22, 2016. The search strategy was based on the following words:  OR “cd147 antigens”[All Fields] OR “cd147”[All Fields]) AND (“stomach neoplasms”[MeSH Terms] OR (“stomach”[All Fields] AND “neoplasms”[All Fields]) OR “stomach neoplasms”[All Fields] OR (“gastric”[All Fields] AND “cancer”[All Fields]) OR “gastric cancer”[All Fields]). Furthermore, we also searched the additional publications from the reference lists of the retrieved articles or reviews which had been previously missed.In this meta-analysis, publications that met the following criteria were selected as candidate articles: (1) inclusion of pathologically confirmed GC patients; (2) investigation of the relationship between the expression of CD147 and GC; (3) available data for calculation of odds ratios (ORs) with their corresponding 95% confidence intervals (CIs). And the following exclusion criteria were used: (1) abstracts and reviews, (2) insufficient data to extract or calculate the ORs, and (3) repeated or overlapping publications.Two authors identified and screened the search findings. A third author reviewed all data entries. The following items were extracted from the eligible studies: name of the first author, publication year, country of the first author, sample size and numbers of cases and controls.2 statistic, where an I2 value from 0 to 25% indicate low heterogeneity, 25-50% moderate heterogeneity and ≥ 50% high heterogeneity. If data was not significantly heterogeneous (P > 0.05 or I2 < 50 %), the pooled effects were calculated using a fixed model, Otherwise, a random-effects model was employed [In this meta-analysis, all statistical analyses were performed using the STATA 12.0 software . Pooled odds ratios (ORs) and 95% confidence intervals (CIs) were calculated to evaluate the strength of the associations. We judged heterogeneity by calculating the Iemployed . PublicaPatients with GC were recruited from a study carried out between 2009 and 2014 at Taihe Hospital, Hubei University of Medicine. Clinic pathologic data for parameters were collected from the pathology report. None of the patients had received radiotherapy or chemotherapy before surgery. From their medical records the data on date of diagnosis, age, sex, tumor size, lymph node metastasis, TNM stage , and the overall survival were extracted. The study has been approved by the institutional review board at Hubei University of Medicine. All methods used in this study were carried out in accordance with the approved guidelines and all experimental protocols were approved by Hubei University of Medicine. Informed consent was obtained from all subjects.After surgical procedures, clinical tissue samples were fixed in 4% paraformaldehyde. Before cutting into 4 μm sections, the samples were embedded in paraffin. The sections were dewaxed and rinsed in 100% xylene for 10 minutes. The sections were then rehydrated in 100% alcohol, 95% alcohol, 90% alcohol, 80% alcohol and 70% alcohol for 5 minutes. Then, 5 % hydrogen peroxide was applied to block endogenous peroxide activity and the samples were washed with phosphate-buffered saline (PBS). Blocking was performed with 5% sodium citrate solution for 15 min at 100 °C. The sections were incubated at 4°C overnight with the anti-CD147 antibody (1:500) . Next day, the slides were washed with PBS and incubated with secondary antibody for 20 minutes at 35 °C, followed by color development with Diaminobenzidine (DAB) for 3 minutes after washed with PBS. The cell nuclei were counterstained with hematoxylin for 3 minutes. Sections were photographed on an Olympus photomicroscope . The degree of IHC staining was evaluated by two independent pathologists. Staining intensity was graded as “0” (negative), “1” (weak), “2” (moderate) and “3” (strong); staining percentage was graded as “0” ( < 5%), “1” (5-25%), “2” (25-50%), “3” (50-75%) or “4” (>75%). A final immunoreactivity scores (IRS) was calculated by multiplying the values of the staining intensity and staining percentage. The IRS value > 4 was defined as high expression and IRS value ≤ 4 as low expression. ImagePro Plus  was used to quantitatively score the tissue sections.The SPSS 19.0 , GraphPad Prism Software 5  and R software (R3.3.0) for windows was used for statistical analysis. Correlation between CD147 expression and clinicopathological characteristics were evaluated by Chi-square test and Fisher's exact tests. Overall survival (OS) rate were estimated by using the Kaplan-Meier method and the log-rank test was used to calculate P Values. Univariate and multivariate Cox regression models were used to calculate hazard ratios and their confidence intervals for the study variables. A one-sided power calculation was performed on the effect size (hazard ratio) of CD147 expression in Cox regression . Histori


Vitis vinifera L. red variety Fetească neagră grown in Romania in 2015 were investigated. Grape pomace extracts total phenolic index, total tannins, total anthocyanins, proanthocyanidins, flavan-3-ol monomers, stilbenes, and DPPH free radical scavenger were measured. The effect of a seven-day pretreatment with grape pomace extracts on the isoprenaline-induced infarct-like lesion in rats was assessed by ECG monitoring, serum levels of creatine kinase, aspartate transaminase, and alanine transaminase. Total serum oxidative status, total antioxidant response, oxidative stress index, malondialdehyde, total thiols, and nitric oxide have been also assessed. Higher phenolic content and antioxidant activity were found in fermented pomace extracts when compared to fresh pomace extracts. Pretreatment with grape pomace extracts significantly improved cardiac and oxidative stress parameters. In conclusion, Fetească neagră pomace extracts had a good in vitro antioxidant activity due to an important phenolic content. In vivo, the extracts had cardioprotective effects against isoprenaline-induced infarct-like lesion by reducing oxidative stress, fresh pomace extracts having a better effect.Grape pomace is a potential source of natural antioxidant agents. Phenolic compounds and antioxidant and cardioprotective properties of fresh and fermented pomace extracts obtained from  Cardiovascular diseases are the first cause of death worldwide. The role of oxidative stress in the pathophysiology of cardiovascular diseases is well documented . Moreove2−), H2O2, and the hydroxyl radical. Initially, they cause mitochondrial depolarization and then activate a positive feedback loop of ROS-induced ROS release , ortho dianisidine, vanadium (III) chloride (VCl3), hydrogen peroxide (HVitis vinifera (L.) var. Fetească neagră  planted in 2006, from Mureş county, Mica parish, part of Târnavelor Plateau , was used in our studies. Grapes were harvested manually at full maturity level, during the 2015 vintage. The GP samples were collected in two winemaking stages: one was supplied immediately after pressing the grapes, the fresh unfermented GP (FNFs), and the other was supplied after 20 days of fermentation at 20°C and must separation, the fermented GP (FNFr). The samples were stored in vacuum bags at −22°C prior to the analysis and were used in the experiments.The grape The GP was submitted to the freeze-drying process and then ground with a domestic blender . The extracts were prepared by adding 50 mL of ethanol 50% and 1 mL of metabisulphite (MBS) 5.25% to 5 g of GP sample. All extracts were placed in a water bath at 50°C for 7 days and stirred daily. Prior to each analysis, the extracts were centrifuged and the supernatants were further used. All samples were processed in 3 repetitions.The total phenolic index (TPI) was determined by measuring 280 nm absorbance of a 1 : 200 dilution of GP extracts . The conThe proanthocyanidins of the GP were extracted and analyzed by acid depolymerization in the presence of an excess of phloroglucinol . The proHPLC identification of GP stilbenes was performed using an Agilent 1200 series system equipped with DAD  and coupled to an AB Sciex 3200 Q TRAP (Applied Biosystems) electrospray ionization mass spectrometry system (ESI-MS/MS) . The chr50  was calculated graphically, using a calibration curve, in the linear range by plotting the extract concentration versus the corresponding scavenging effect (RSA%), over 30 min. Trolox was used as a positive antioxidant control. The percentage of DPPH consumption was converted to trolox equivalents (TE) using a calibration curve (R2 = 0.985) of Trolox standard solutions (0.5–5 μg/mL). An IC50< 50 μg TE/ml is a very good antioxidant activity; an IC50 of 50–100 μg TE/ml is a good antioxidant activity; an IC50 of 100–200 μg TE/ml is a week antioxidant activity; an IC50> 200 μg TE/ml means no antioxidant activity. Assay was performed in triplicate.The antioxidant capacities of the GP samples were measured in terms of their radical scavenging activity (RSA), using the DPPH method , 23. BriFetească neagră fresh GP extract (FNFs) and Fetească neagră-fermented GP extract (FNFr) were obtained with 70% ethanol  by a modified Squibb repercolation method (1/1 g/ml) .The experiments were carried out on male albino Wistar rats, weighing 200–250 g, that were bred in the Animal Facility of the Iuliu Hațieganu University of Medicine and Pharmacy. The animals were housed in standard polypropylene cages (five per cage) under controlled conditions  and with ad libitum access to standard pellet diet  and water.Experimental protocols have been approved by the Ethics Committee (nr. 26/16.12.2015) of the Faculty of Veterinary Medicine, University of Agricultural Sciences and Veterinary Medicine from Cluj-Napoca and the Ethics Committee of the Iuliu Hațieganu University of Medicine and Pharmacy. The experiments were performed in triplicate. All experimental groups began with 3 days of acclimatization to the housing facility, and animals were used only once. At the end of the experiments under anaesthesia using a combination of ketamine (60 mg/kg bw) and xylazine (15 mg/kg bw) , blood wn = 5). The negative control group (CONTROL) and the isoprenaline-induced (ISO) myocardial infarction group received 0.9% saline (1 ml/day p.o.) for 7 days. In FNFs and FNFr groups, each extract was administrated orally by gavage (1 mL/day p.o.) for 7 days. In day 8 and day 9, rats were injected with ISO dissolved in normal saline  at an interval of 24 h to induce experimental MI [The animals were divided into 4 groups  and xylazine . At 15 min after anesthesia, animals were placed in the supine position on a board, electrodes were bound on the paw pads of each rat, and ECG was recorded from the limb lead at position II (right forelimb to left hind limb) with a Biopac MP150 system. The ECG apparatus was calibrated at 1 mV/1 cm with a speed of 50 mm/s.Analysis of ECG waves was done to calculate heart rate (beats/min), RR intervals (msec), QT interval (msec), and ST segment changes (mV). QT interval was measured from the beginning of QRS complex to the end of T wave, and it was calculated in msec. Corrected QT interval (QTc), which is used to rectify the influence of the heart rate on QT interval, according to Bazett formula, is equal to QT interval divided by the square root of RR interval and was also calculated .The creatine kinase-MB (CK-MB), aspartate transaminase (AST), and alanine transaminase (ALT) activities were measured by using commercial kits.2O2) as the oxidative species, and the assay results are expressed in μmol H2O2 equiv/L.The total oxidative status (TOS) of the serum was measured using a colorimetric assay . Assay mThe total antioxidant response (TAR) was measured in serum using a colorimetric assay . This as2O2 equiv/L)/TAR (mmol trolox equiv/L) [The ratio of the TOS to the TAR represents the oxidative stress index (OSI), an indicator of the degree of oxidative stress: OSI (arbitrary unit) = TOS (mol Hequiv/L) .Malondialdehyde (MDA) was assessed as a lipid peroxidation marker, using thiobarbituric acid, as previously described . The absTotal thiols (SH) were estimated using Ellman's reagent . Supernav : v) solution of methanol/diethyl ether [μmol/L [The Griess reaction was used to indirectly determine NO synthesis (NOx). First serum proteins were removed by extraction with a 3 : 1 .r) was used to evaluate relationships between parameters of the same group. The level of significance was set at p < 0.05.Results were expressed as means and standard deviations and were analyzed statistically using the SPSS (version 20) software. Data were compared by one-way ANOVA and post hoc Bonferroni-Holm test. Pearson's correlation coefficient (p < 0.05). TC in GP ranged from 63.15 ± 7.12 mg epicatechin/g d.w. in FNFs to 113.98 ± 4.39 mg epicatechin/g d.w. in FnFr, the difference being very significant (p < 0.001). Higher TAC was found in FNFs  than in FNFr . Between TAC of FNFs and FNFr, the difference was highly significant (p < 0.001).The TPI, TAC, and TC obtained from Fetească neagră GP are shown in p < 0.05). As shown in p > 0.05), PD% in FNFs was significantly bigger than that in FNFr (p < 0.01), and G% in FNFr was higher than that in FNFs but not significant (p > 0.05).The results of analyzing GP proanthocyanidins obtained by acid depolymerization in the presence of excess phloroglucinol are shown in p < 0.01). The analysis of individual flavan-3-ol monomers showed that the difference in total monomers concentrations was due to the higher content of (+)-catechin and (−)-epicatechin in the FNFr samples (p < 0.01). The epicatechin-3-O-gallate concentration was not significantly different between the FNFr and FNFs samples (p > 0.05).The flavan-3-ol monomeric compositions from the whole Fetească neagră GP were described in a detailed form in p < 0.01) than in the FNFr and cis-resveratrol in a higher amount in the FNFr (p < 0.01) than in the FNFs. In both FNFs and FNFr samples, the concentration of cis-resveratrol was higher than the concentration of transresveratrol. Piceid was the most abundant stilbenes in GP from both FNFs and FNFr, and both isomers cis- and transpiceid were in a greater quantity in the FNFs samples as compared to the FNFr samples (p < 0.001) (The transresveratrol was in a higher amount in the FNFs (< 0.001) .50 = 11.18 μg/ml), FNFr (IC50 = 9.95 μg TE/ml) exhibits a better antioxidant activity than FNFs (IC50 = 36.99 μg TE/ml), and there was a statistically significant difference between these extracts (p < 0.001). For FNFs, DPPH test results exhibited significant, positive correlations with TC, TAC, flavan-3-ol monomers, and stilbene results (r = 0.80–0.99). The FNFr DPPH test correlated with proanthocyanidins, PD, mDP, and cis-stilbenes (r = 0.76–0.90).The ethanol extracts of FNFs and FNFr showed very good RSA by having an increased absorbance with increased concentration at 517 nm . CompareECG recording from days one and 7 had no significant changes in all groups. The ECG patterns of normal and experimental group rats in day 10 are shown in p < 0.001). FNFs and FNFr extract pretreatment produced protective effects by reducing significantly AST, ALT, and CK-MB levels. On AST, FNFs effect (p < 0.01) was better than that of FNFr (p < 0.05). ALT was significantly reduced by both pretreatments, FNFs and FNFr (p < 0.01), and the results were almost similar. FNFr lowered CK-MB more than FNFs did (p < 0.001). The cardioprotective effects of FNFs and FNFr on cardiac enzymes were correlated with ST segment changes.The cardioprotective effects of Fetească neagră GP extracts revealed by the serum cardiac marker enzyme AST, ALT, and CK-MB levels are summarized in p < 0.001) and TOS (p < 0.001) with concomitant decrease in TAR (p < 0.01) were seen in the ISO-induced MI rats. In contrast, individual preadministration of FNFs and FNFr resulted in a significant decrease in the levels of TOS (p < 0.001) and OSI (p < 0.001) as well as significant increases in TAR (p < 0.01). However, when FNFs and FNFr groups were compared to each other, the ANOVA analysis revealed that the decrease in TOS and OSI levels and the increase in TAR were not significantly different. Furthermore, OSI, TOS, and TAR changes were correlated. ISO-induced ischemia was associated with a significant elevation of MDA (p < 0.01), and FNFs and FNFr pretreatments reduced MDA production (p < 0.05). FNFs and FNFr were equally efficient on MDA (p > 0.05). SH was reduced by the ISO-induced ischemia (p < 0.001), and pretreatment with FNFs and FNFr extracts had no important effect on SH (p > 0.05). Serum NOx was significantly elevated in the ISO group (p < 0.001) as compared to the control rats. Pretreatment with FNFs and FNFr extracts induced no significant reduction of nitric oxide production (p > 0.05). There was no significant statistical difference between FNFs and FNFr extracts in terms of NOx production (p > 0.05).Oxidative stress tests are summarized in In the present study, polyphenolic composition and antioxidant and cardioprotector effects of fresh and fermented FN GP were analysed. The FNFs and FNFr demonstrated to be rich sources of phenolic compounds. It is already known that phenolic compounds represent one of the largest and important groups of natural plant products, having physiological properties such as antiallergenic, anti-inflammatory, antimicrobial, antioxidant, antithrombotic, cardioprotective, and vasodilatory effects . There iAs in other studies, FNFs and FNFr phenolic contents were high . The difTannins are localized mainly in the skin and seeds of grapes. Tannin content in FN GP followed the same trend as TPI, FNFr having a higher tannin yield than FNFs. These results were in accordance with reports that found tannin extraction efficiency from unfermented grape skins in a hydroalcoholic solution lower than 38% . When saBetween different cultivars, there are differences of TAC . MoreoveThe total proanthocyanidin of wines is affected by maturity and maceration. Proanthocyanidin concentration is higher at veraison, decreases until ripeness, and then remains relatively constant. Then, a longer maceration increases the proanthocyanidin content . By puttProanthocyanidin structures differ according to their origin. In wines, the proanthocyanidin mean degree of polymerization (mDP) and the percentage of PD increased with maturity, whereas the G% decreased. Furthermore, maceration length also has a significant effect on the proanthocyanidin mDP of wines. Throughout maceration time, the proportion of PD tends to decrease , but theFlavan-3-ols are extracted from grapes during the vinification process, but not completely, remaining in great amounts in the winemaking residue. Like in other studies, analysing red GP flavan-3-ol monomers , in FN Gtrans- and cis-piceid, piceatannol, and resveratrol dimers (viniferins) [in vitro studies have shown that resveratrol has antioxidant, anti-inflammatory, cardioprotective, and platelet antiaggregant activities, and glycosylated stilbenes have antioxidant effects [trans- and cis-isomers as previously described for other red GP [cis-form [Stilbenes are among the main polyphenols involved in the health protection effects of drinking wine. The main grape stilbenes are trans- and cis-resveratrol, glycosylated derivatives of resveratrol iferins) . Several effects . Signifi effects  and betw effects . In FNFsr red GP , 51. Totr red GP , but FNFcis-form . Becausecis-form  and may cis-form , high piin vitro antioxidant activities were evaluated with the most frequently used method, the DPPH test [50 than FNFs. This observation is consistent with other studies indicating that the fermentation process increases polyphenol releases from the pomace cells [in vitro antioxidant activities and TPI was found. The different structure of each phenolic compound explains their specific efficiency to scavenge different free radicals [An actual trend is to reduce oxidative stress by using herbal supplements or functional foods rich in natural antioxidants. FNFs and FNFr PPH test . The higce cells . As phenradicals .in vitro and in vivo antioxidant effects of a plant extract have shown that some extracts exhibit antioxidant activity both in vitro and in vivo, but for other extracts, the in vitro antioxidant activity does not apply to in vivo models [in vivo cardioprotective and antioxidant effects in experimental rat ISO-induced MI [The antioxidant capacity of plant extracts cannot be evaluated by using a single test. Studies comparing the o models . Due to duced MI .When injected in rats, isoprenaline undergoes autooxidation and induces myocardial cell membrane injury . ISO indMyocardial injury leads to loss of cell membrane structural integrity, with increased permeability and enzyme leakage. The degree of AST, ALT, and CK-MB increase in serum reflects the severity of myocardial injury . Like inThe myocardium is vulnerable to oxidative damage because it has limited antioxidant defence possibilities . In MI, in vivo activity. The antioxidant activity of plant extracts is the result of the combined effects of many different compounds. It was demonstrated that the antioxidant in vitro activity does not always apply to in vivo models. Moreover, it was proved that in vitro and in vivo antioxidant effects may not correspond because polyphenols may also act as prooxidants via Fenton reaction [In the present study, oxidative stress was evaluated systemically by measuring serum oxidative stress markers. As a result, an increased oxidative stress, characterized by the TOS, OSI, and MDA elevation associated with TAR lowering, has been found. Both FNFs and FNFr had cardioprotective effects by reducing oxidative stress markers, but FNFs had better reaction .Nitric oxide (NO) is an important signalling messenger known to play important roles in many physiological and pathological conditions. Endogenous NO is generated from L-arginine by three major nitric oxide synthases: endothelial, neural, and inducible nitric oxide synthases (iNOS). NO plays an important role in host defence and homeostasis when generated at a low level for a brief period of time, whereas the high level or prolonged induction of NO may contribute to a variety of pathological phenomena associated with inflammatory processes . In the in vitro antioxidant activity due to an important phenolic content. In vivo Fetească neagră grape pomace extracts had cardioprotective effects against ISO-induced myocardial ischemia by reducing oxidative stress. The in vitro and in vivo antioxidant effects were different, respectively, fermented Fetească neagră grape pomace had a better in vitro antioxidant activity, and fresh Fetească neagră grape pomace had a stronger in vivo antioxidant activity. Overall, the findings presented in this study suggest for the first time that Fetească neagră grape pomace extract pretreatment may be an option for heart preconditioning. Further studies are required to investigate the cardioprotective effects of Fetească neagră grape pomace extracts over a longer period.In conclusion, fresh or fermented Fetească neagră grape pomace extract had a good


Polyphenols are antioxidant molecules found in many foods including nuts, fruits, vegetables, chocolate, wine, and tea. Polyphenols have antimicrobial, anti-inflammatory, and antineoplastic properties. Recent studies suggest that tea polyphenols may be used for reducing sebum production in the skin and for treatment of acne vulgaris. This review examines the evidence for use of topically and orally ingested tea polyphenols against sebum production and for acne treatment and prevention. The PubMed database was searched for studies on tea polyphenols, sebum secretion, and acne vulgaris. Of the 59 studies found, eight met the inclusion criteria. Two studies evaluated tea polyphenol effects on sebum production; six studies examined tea polyphenol effects on acne vulgaris. Seven studies evaluated topical tea polyphenols; one study examined systemic tea polyphenols. None of the studies evaluated both topical and systemic tea polyphenols. Tea polyphenol sources included green tea (six studies) and tea, type not specified (two studies). Overall, there is some evidence that tea polyphenols in topical formulation may be beneficial in reducing sebum secretion and in treatment of acne. Research studies of high quality and with large sample sizes are needed to assess the efficacy of tea polyphenols in topical and oral prevention of acne vulgaris and lipid synthesis by the sebaceous glands. Polyphenols are naturally occurring compounds found in plants and common foods such as nuts, fruits, vegetables, chocolate, wine, and tea. After ingestion, polyphenols are absorbed in the intestinal epithelium  and haveCamellia sinensis is the plant that gives rise to a variety of teas depending on specific processing of the plant [Tea is the second most consumed beverage worldwide and an important source of plant polyphenols in the human diet . Camellihe plant . Green the plant . Componehe plant . Catechihe plant .Propionibacterium acnes (P. acnes); and (d) inflammation around pilosebaceous follicles [Sebaceous glands are found throughout the human body, especially on the face and scalp, and they produce sebum—a mixture of lipids . Lipids ollicles ,17.Recent evidence suggests that there is increased phosphoinositide 3-kinase-Akt-mammalian target of rapamycin complex 1 (PI3K-Akt-mTORC1) signaling in the skin of patients with acne vulgaris ,19,20 anThere are multiple acne grading systems but there is no gold standard  that is P. acnes and reduces inflammation caused by P. acnes. The aim of this review is to examine recent clinical trials on tea polyphenols and their effects on sebum production and acne vulgaris management.Due to the complex nature of acne vulgaris, most treatments have only modest efficacy . SystemiOn 4 August 2016 the PubMed database was searched for studies that investigated tea polyphenols, sebum production, and acne. No limits were placed on the publication date. The search combined keywords “tea”, “green tea”, “EGCG”, “polyphenol”, “catechin”, “sebum” and “acne”. No filters were selected.Records were screened by title and/or abstract to exclude studies that did not contribute to answering the question in this review. Inclusion criteria: (1) published in English; (2) intervention included a tea polyphenol; (3) human study. Exclusion criteria: (1) in vitro studies; (2) animal studies; (3) review articles.n, age); (6) study design, duration; (7) major outcome measures; (8) major results; (9) adverse events; (10) reference.Data was extracted from selected studies  as folloOur search yielded a total of 59 studies. Titles and/or abstracts were screened for inclusion and exclusion criteria. Eight articles met the criteria and were subsequently analyzed. Two studies examined the effects of tea polyphenols on sebum production, while six studies investigated the effects of tea polyphenols on acne vulgaris. Seven studies examined the application of topical tea polyphenols and one study examined systemic tea polyphenols. None of the studies examined both topical and systemic tea polyphenols. Tea polyphenol sources included green tea in six studies, while in two studies the tea type was not specified.n = 11). The second group used topical green tea and lotus (GT-L) on one cheek and placebo control on the other (n = 11). The GT topical consisted of 5% green tea and the GT-L formulation contained 2.5% green tea and 2.5% lotus extract. Both groups were instructed to apply their respective topicals at bedtime for 60 days. Sebumeter was used to measure sebum secretion on both cheeks at baseline and days 15, 30, 45, and 60. After 60 days, the GT topical group had a significant reduction in sebum secretion from baseline, compared to placebo control . The GT-L group also had a significant reduction in sebum secretion from baseline to day 60 compared to placebo control . There were no adverse events reported in either GT or GT-L group. This study showed that GT and GT-L topicals were both effective in significantly reducing sebum secretion. Additionally, even though the GT-L combination topical had a smaller percentage of GT extract (2.5% vs. 5% in GT group), GT-L reduced sebum secretion to a higher degree than GT topical alone; this could indicate an additive effect of GT and L extracts. These findings suggest that GT and GT-L combination topicals could be used to treat skin diseases that are associated with increased sebum secretion, such as acne vulgaris. Limitations of this study include a small sample size and that it only included healthy men. Future studies should focus on studying the effects of the two formulations in patients with acne vulgaris.In a single-blinded, placebo controlled monocentric study by Mahmood et al., 2013  conductep < 0.05). Specifically, sebum production decreased by nearly 10% in the first week and as much as 60% by week 8. No side effects were noted. Limitations of this study include small sample size, no comparison treatment or placebo control, and it did not test individuals with skin diseases known to have high sebum production.In a study by Mahmood et al., 2010 , ten hean = 24) or 5% zinc sulphate solution (n = 23); both groups applied their respective topical twice daily for 2 months. Inflammatory lesion counting was performed every 2 weeks and graded as follows: mild acne (<20 pustules and <10 papules) and moderate acne (20–40 pustules and >30 papules). It was considered that subjects had a “good response” to treatment if the inflammatory lesion count decreased by >50%; “moderate response” when decrease was 10%–50%; and “no response” when lesion count decreased by <10%. The 2% tea lotion group experienced a significant reduction in inflammatory lesion count . Eighty-five percent of subjects had a good or moderate response and 15% had no response to treatment. There was no significant difference in inflammatory lesion count (papule or pustule) in the group treated with 5% zinc sulphate compared to baseline. Adverse event reported by the 2% tea lotion group was mild itching (5 subjects) at the beginning of the treatment period. The 5% zinc sulphate group also reported itching (2 subjects), as well as burning sensation (5 subjects). No serious side effects were reported in either group. One limitation of this study is the small sample size (n = 20 in each group that completed the study). Nevertheless, 2% tea lotion is promising for its effect of decreasing inflammatory lesion count. Future studies could be done to compare it to other known topical treatments for acne to further assess its efficacy and safety profile.Sharquie et al., 2008  conductep < 0.05). Specifically, non-inflammatory lesions reduced by 79% and inflammatory lesions by 89% after eight weeks of 1% EGCG use compared to baseline (p < 0.05). The non-inflammatory and inflammatory lesion reduction was similar in 5% EGCG group (p < 0.05), however, the specific lesion counts and percentage improvement were not provided. Overall, this study showed that 1% and 5% EGCG were both effective in improving acne vulgaris without major side effects.A randomly-assigned split body (face) trial was conducted by Yoon et al., 2013  to examip < 0.001; baseline pustules 20.7 ± 5.8, week 8 pustules 8.9 ± 2.3, p < 0.001). Eighty-eight percent of subjects in the treatment group reported being fully or partially satisfied with the treatment outcome and 12% were not satisfied. The control group did not have a significant reduction in lesion count at 8 weeks compared to baseline. In this group, 83.3% of subjects were not satisfied with the outcome and 16.7% were partially or fully satisfied. There were no adverse events reported. This study suggests that 2% tea lotion is an effective topical alternative in acne management.A single-blind randomized controlled study by Sharquie et al., 2006  investigp < 0.0001, 95% confidence interval (CI) = 8.58–19.42). The mean SI also decreased from 2.05 (baseline) to 1.25 after 6 weeks of treatment . Five subjects reported stinging sensation or itching, but these resolved within 2–3 days of treatment. Small sample size and lack of comparison, or control group, were limitations in this study. The true effect of the topical green tea lotion was not clear with the lack of a control treatment. Nevertheless, it showed that 2% green tea lotion could be beneficial in acne treatment by reducing acne lesion count and warrants future controlled studies.Elsaie et al., 2009  studied p < 0.05) and 28% decrease in the number of pustules (p < 0.05) compared to baseline. There was no significant decrease in the number of closed comedones at week 8 compared to baseline. No major adverse events were noted. This study did not have a control group or control treatment.In a study by Jung et al., 2012  conducten = 40) received 1500 mg decaffeinated green tea daily  for four weeks. The second group received cellulose capsules (placebo), which looked identical to GT capsules for blinding purposes. A blinded dermatologist recorded inflammatory and non-inflammatory lesion counts on the forehead, cheeks, nose, perioral area, chin, and entire face at baseline and at four weeks post-treatment. There was a significant decrease in the inflammatory lesion counts on the nose (p = 0.03), perioral area (p = 0.04), and chin (p = 0.03) in the green tea group compared to placebo group. There was no significant difference in lesion counts on forehead, cheek, or in the total lesion count between GT and placebo groups. Within the GT group, subjects had significantly lower inflammatory lesion counts on the forehead (p = 0.04) and cheeks (p = 0.04), as well as lower total lesion count (p = 0.03) at the end of treatment compared to baseline. Within the placebo group, there was significant decrease in inflammatory lesion count on cheeks (p = 0.01) and chin (p = 0.01), as well as total lesion count (p = 0.02). Adverse events included mild constipation and abdominal discomfort (three subjects). Placebo group adverse events included thirst and difficulty falling asleep (two subjects). However, no major side effects were reported. Even though decaffeinated GT treatment resulted in decreased lesion counts on nose, perioral area, and chin compared to placebo, the placebo group itself had significant within group difference in lesion counts before and after treatment. Additional studies with more subjects and longer duration are needed to assess the effectiveness of systemic green tea in treatment of acne.Lu et al., 2016  conducteCamellia sinensis [Polyphenols are naturally occurring compounds found in many common foods and plants, including sinensis , the plasinensis , and thiBioavailability of polyphenols varies depending on the polyphenol forms within the dietary source . EGCG, tThe partition coefficients of tea polyphenols are shown in Overall, studies show that topical and systemic tea polyphenols tend to be well tolerated. A study by Elsaie et al.  showed mPropionibacterium acnes (P. acnes), sebum production by the sebaceous gland, keratinization of the follicular keratinocytes, and inflammation [P. acnes [Propionibacterium granulosum [Staphylococcus aureus [Staphylococcus epidermidis [P. acnes, P. granulosum, S. aureus, and S. epidermidis was inhibited at concentrations of 400 µg gallic acid equivalents (GAE)/mL GT or less.Acne is thought to be due to the activity of bacteria called ammation . VariousP. acnes , Propionanulosum , Staphyls aureus  and Stapdermidis  are bactdermidis  suggestsP. acnes. Altogether, these findings further support the notion that EGCG has anti-microbial activity against P. acnes. EGCG was also found to reduce transcription of the nuclear factor-κB (NF-ĸB) pathway and reduce inflammation. Overall, studies suggest that tea polyphenols exert their effect on sebum production and acne via several mechanisms, including acting as anti-microbial, anti-lipogenic, anti-inflammatory molecules.In addition to the anti-microbial properties of tea polyphenols, other mechanisms have been proposed to explain the improvement in acne from topical green tea administration. Yoon et al., 2013  postulatInterpretation of this review should be considered in light of the limited number of studies that were available. Many of the studies had small sample sizes and some studies had poor research design, including a lack of control group. More studies of high quality are needed to establish the efficacy of tea polyphenols for controlling sebum production, as well as preventing or managing acne vulgaris.Our search produced a limited number of results for how tea polyphenols may be used in reducing sebum secretion and acne pathology. While all the studies used some form of tea polyphenols, the exposures were not uniform. Some studies included unknown types of tea and with unspecified amounts of tea polyphenols as exposure. However, based on this review there is some evidence that tea polyphenols used orally and topically may be beneficial for skin health and more specifically, for reducing sebum production by the sebaceous glands and for the prevention and treatment of acne vulgaris. Physicians and other health care professionals should be aware of the studies examining the beneficial effects of tea polyphenols as they could potentially be used as alternatives in skin care. Research studies of high quality and with large sample sizes are needed to assess the efficacy of tea polyphenols in topical and oral prevention of acne vulgaris and lipid synthesis by the sebaceous glands.


Toxic shock syndrome is a potentially fatal toxin-mediated disease. The role of toxins in this clinical entity made us hypothesize that extracorporeal blood purification with CytoSorb® could play a beneficial role in the clinical management of toxic shock syndrome. This case report describes the successful treatment of toxic shock syndrome using a combination of renal replacement therapy and hemadsorption in a pediatric patient. A 5-year-old girl with Down's syndrome presented with an inflamed area surrounding an insect bite, signs of systemic inflammation, and multiple organ failure. As previous attempts of immune modulation therapy were unsuccessful, renal replacement therapy was supplemented by the cytokine absorber CytoSorb. Treatment using this combination was associated with a rapid and significant stabilization in the hemodynamic situation and a decrease in inflammatory mediators within hours after the initiation of therapy. The application of CytoSorb therapy was simple and safe. The use of extracorporeal blood purification with CytoSorb proved potentially beneficial by removing toxins and inflammatory mediators in this case and could therefore play a role in the clinical management of toxic shock syndrome. Whether CytoSorb has the potential to even positively influence mortality in patients with toxic shock syndrome still needs to be confirmed. Staphylococcus aureus or Group A Streptococcus (GAS). This multisystem disorder is the consequence of an uncontrolled systemic inflammatory response syndrome caused by bacterial superantigens that broadly activate T-cells and lead to hypercytokinemia and hyperinflammation [Toxic shock syndrome (TSS) is a potentially fatal toxin-mediated disease caused by infection fromammation . Consequ S. pyogenes or S. aureus/epidermidis the day before admission by the GP. Despite antibiotics, the erythema continued to migrate proximally towards the inguinal region. On arrival at the emergency department, the conscious patient presented with diarrhea but stable vital signs: breathing rate 26 breaths/min, heart rate 110 beats/min, and blood pressure 98/57 mmHg. Blood examination revealed leukocytosis (15.44 G/L), neutrophilia , mild anemia , a normal platelet count (238 G/L), and an elevated CRP (260.98 mg/L).A 5-year-old girl with Down's syndrome presented with an erythematous, indurated area on her right leg surrounding an insect bite noticed two days prior to admission. Oral cefaclor therapy was started for suspected systemic infection by2 37.6 mmHg, HCO3 5.2 mmol/L, and lactate 7 mmol/L. Septic shock was diagnosed and three intravenous fluid boluses (20 mL/kg) and oxygen therapy were started. Blood cultures were taken and the antibiotic therapy augmented by parenteral clindamycin and vancomycin. Due to an unsatisfactory cardiovascular response to fluid resuscitation, dopamine was started at 5 μg/kg/min and then increased to 10 μg/kg/min. Low ventricular contractility on ultrasonography required additional dobutamine at 5 μg/kg/min while clinical signs of decreased systemic vascular resistance necessitated noradrenaline at 0.1 μg/kg/min which had to be increased up to 0.5 μg/kg/min later. Due to pronounced cardiovascular instability and suspected adrenal gland insufficiency, hydrocortisone was prescribed at 2 mg/kg/12 hours.During the first two hours her status deteriorated so rapidly that she needed to be transferred to the intensive care unit. Capillary blood gas analysis revealed a severe uncompensated metabolic acidosis: pH 6.77, pCO S. pyogenes or S. aureus was given on the day of admission and the following day. Fresh frozen plasma (15 mL/kg) and low molecular weight heparin (enoxaparin 6 mg/kg) were administered for laboratory signs of disseminated intravascular coagulation (DIC). Deep vein thrombosis of the right lower extremity was ruled out by Doppler ultrasonography. Chest X-ray revealed increased perihilar lung markings.Considering the well-known immunodeficiency of patients with Down's syndrome, an antimycotic agent (fluconazole) was administered preventively, and intravenous immunoglobulin therapy (1 g/kg) for septic shock due to2O, PEEP 5 cm H2O, respiratory rate 25/min, and FiO2 0.4.Eight hours after admission to the ICU an altered state of consciousness, hypercapnia, tachydyspnoe, and increasing oxygen needs required intubation and pressure controlled SIMV at PIP 17 cm HBased on the Surviving Sepsis Campaign guidelines multiorgan failure was diagnosed. The patient had cardiovascular instability requiring complex vasoactive therapy ; DIC indicated deteriorating liver function, poor lung function on ventilatory support alluding to Acute Respiratory Distress Syndrome (ARDS), and deteriorating kidney function. Microbiological examinations were inconclusive as to the causative pathogen.During the first 48 hours of therapy diffuse erythroderma with purpura and petechiae appeared, while the right leg became livid, and bullae appeared in places subjected to external pressure . PeriodiIn an attempt to modulate the dysregulated immune response, plasma exchange therapy was performed without complications. Despite an increasing urine output during the first 24 hours from 0.4 to 5.4 ml/kg/hour her fluid balance remained positive. Thereafter, diuresis declined and the patient became anuric by hour 56 such that continuous venovenous hemodialysis was started. Thrombocytopenia, anemia, and hypoproteinemia were supported by blood products and an augmentation of the low neutrophil levels by granulocyte-colony stimulating factor tried on days four and five.Qb) of 40 mL/min could be achieved; dialysis fluid (MultiBic) flow (Qd) of 800 mL/hour was maintained, with an ultrafiltration rate of 100 mL/hour. Because of hepatic failure with DIC, regional citrate anticoagulation was used during the whole course of CRRT without any remarkable complication. As a result of a 72-hour uneventful course of the CVVHD-CytoSorb therapy, cardiovascular stability improved on lower vasopressor doses; FiO2 decreased from 0.95 to 0.4, airway pressures PIP/PEEP from 27/10 to 24/8 cm H2O, and respiratory rate from 25 to 15/min, respectively. In addition, levels of important laboratory parameters decreased . CVVHD was performed with multiFiltrate  machine, using a AV400S polysulfone dialyzer. Blood flow , damage-associated molecular pattern molecules (DAMPs), and cytokines from whole blood in a single compartment in vitro recirculation system provided that substantial quantities of a broad spectrum of DAMPS, PAMPS, and cytokines  were removed [The clinical picture in our case was ambiguous. The cutaneous lesion, as probable origin of the disease, suggested removed . Therefoγ, CRP, and PCT but also of metabolic parameters  started to decrease rapidly. This attenuation of the inflammatory response correlated well with the clinical improvement of the patient.One of the striking features of systemic TSS is the speed of excess cytokine and toxin production. Our patient had been treated with multiple immune-modulatory therapies  achieving only moderate reductions in the laboratory parameters that characterize systemic inflammation. However, from the start of CVVHD/CytoSorb initiation, plasma levels of inflammatory mediators such as IL-6, IL-10, IFN-Previous studies have shown that inflammatory mediators can be removed from the circulation with different blood purification techniques such as plasma exchange, high-volume and very high-volume hemofiltration, and a number of hybrid therapies encompassing high-permeability hemofiltration, super high-flux hemofiltration, hemadsorption or coupled filtration, and adsorption . These tHowever, despite the fact that most experience with CytoSorb in the clinical setting to date is still limited to case reports, the first multicenter randomized controlled study using treatment with CytoSorb hemadsorption in septic patients with acute lung injury showed that application of the device over a 7-day treatment period was safe and effectively decreased the blood concentration levels of several key cytokines –11. FromThe use of extracorporeal blood purification with a novel sorbent, CytoSorb, proved potentially beneficial by removing toxins, cytokines, and other inflammatory mediators in this case of toxic shock syndrome in a pediatric patient and could therefore play a future role in the clinical management of toxic shock syndrome. Whether CytoSorb has the potential to even positively influence mortality in patients with toxic shock syndrome needs to be confirmed in randomized, controlled trials.


TaqI, ApaI, FokI and BsmI) have been reported to influence Hashimoto’s thyroiditis (HT) risk. However, individual studies have produced inconsistent results. We conducted a comprehensive meta-analysis of eleven case-control studies to better understand roles of the four polymorphisms in HT development. The results showed only FokI polymorphism was significantly associated with the risk of HT . Subgroup analyses demonstrated the significant effect was only present in Asian population , but not in Caucasian. For TaqI, ApaI and BsmI polymorphisms, no significant association was found in any model comparison. Based on the current literature, it appears that only VDR FokI polymorphism is associated with HT risk in Asian population, but not in Caucasians; and the TaqI, ApaI and BsmI polymorphisms have not positive association neither in the overall population, nor when stratified by ethnicity. Further well-designed studies with larger sample sizes and different ethnic population are needed to clarify the present findings.Four VD receptor (VDR) gene polymorphisms ( Hashimoto’s thyroiditis (HT) is an autoimmune thyroid disease (AITD), which has been reported to lead hypothyroidism in up to 5% of population12457810121314Vitamin D receptor (VDR) is a ligand inducible transcription factor, which is harbored on many human immune cells16171920222124252628293031323334353637Therefore, it is necessary to carry out a meta-analysis of the available evidence to clarify this inconsistency and provide a much comprehensive and quantitative understanding of the association of VDR gene polymorphisms with HT risk.P-value of chi square test for genotype distribution and HWE in control group. HT is often diagnosed mainly on the basis of laboratory and ultrasonographic features, such as positive serum anti-thyroid antibodies, heterogeneous echo-structure with diffuse or patchy hypoechogenicity at ultrasonography, with hypothyroid or euthyroid metabolic state.As shown in 2829303132333435363728293132343033353728P-value of Egger’s test for publication bias.FokI polymorphism and HT risk. Pooled analyses showed a significant association in the allele model  and the dominant model , but not in the other models . Then, Galbraith plot analyses were performed to further explore the sources of heterogeneity. As shown in et al.et al.I2 = 0% and P = 0.760 for F vs f; I2 = 0% and P = 0.738 for FF vs Ff + ff) while the overall association remained significant in these two models  (P = 0.030), but the P value of the dominant model was borderline . Subgroup analyses by ethnicity indicated that the FokI F allele or FF genotype significantly increased the risk of HT in Asians , but the positive association was not found in Caucasians. However, significant heterogeneity were also detected in two models among studies with Asian population  , with no significant heterogeneity . Subgroup analyses by study quality suggested that this positive association only existed in pooled analyses of high-quality studies .Eight studies including 978 cases and 938 controls examined the association of r models , Fig. 2.= 0.000) . There w= 0.020) . GalbraiBsmI polymorphism and HT risk. Pooled results indicated that there was no significant correlation between BsmI polymorphism and HT risk in all genetic models  in the overall population (31I2 = 52.1% and P = 0.064). A Galbraith plot analysis suggested that Stefanic et al.P = 0.615), with no significant heterogeneity (I2 = 0% and P = 0.621).Six studies including 837 cases and 901 controls evaluated the association of pulation . Similarpulation . MoreoveApaI polymorphism and HT risk. The meta-analyses demonstrated no positive relationship of ApaI polymorphism and HT risk in the overall population . No significant heterogeneity was found in all the comparisons .Six studies including 766 cases and 813 controls evaluated the association of ulations . SensitiTaqI polymorphism and HT risk. The genotype distribution was consistent with HWE in the controls of all studies . In the Galbraith plots, two studies33I2 = 0% and P = 0.635 for heterogeneity; TT vs Tt + tt: OR = 1.16, 95% CI = 0.90–1.50, P = 0.262; I2 = 0% and P = 0.788 for heterogeneity). Subgroup analyses by ethnicity found the similar results in Caucasian or in Asian   .FokI, TaqI and ApaI polymorphism . However, significant publication bias was detected in two models regarding BsmI polymorphism (PEgger’s = 0.001 for Bb vs bb and PEgger’s = 0.005 for BB + Bb vs bb) . The imputed studies produced symmetrical funnel plots  . We usedel plots .BsmI and TaqI polymorphisms and AITD risk has been reported by a previous meta-analysis2953433343536FokI but not TaqI, ApaI and BsmI polymorphism was significantly associated with the risk of HT. Furthermore, the positive association of FokI polymorphism was only detected in Asians, not in Caucasians by subgroup analyses based on ethnicity.To our knowledge, this is the first meta-analysis specially focused on the association of VDR polymorphism with HT risk. A significant association between the FokI (rs2228570), located in the translational initiation site of VDR, which is the only known VDR gene polymorphism that results in the generation of an altered protein414244FokI polymorphism in patients with HT and controls, and six found a positive association, but another two studies36P = 0.010) and the incidence of HT was significantly higher in FF genotype individuals than that of Ff + ff genotype individuals in overall population . In addition, results from subgroup analyses stratified by ethnicity indicated that HT risk was increased in Asians with FF genotype , but not in Caucasians. This inconsistent result in these two ethnicities may be due to the influence of different genetic backgrounds, lifestyle and environment factors (such as sunlight exposure and diet). In addition, an insufficient number of samples for analysis might lead to unreliable conclusions with deviation in Caucasians.Polymorphism BsmI (rs1544410), ApaI (rs7975232), and TaqI (rs731236) SNPs, located near the 3′ end of the VDR gene, are in strong linkage disequilibrium (LD) with each other. These three SNPs don’t change the amino acid sequence of the encoded protein but have been shown to affect gene expression through regulation of mRNA stability3033TaqI polymorphism was associated with risk of HT in Croatian and Turkish population, but four other studies3435ApaI polymorphism was reported no association with HT risk in previous studies with consistent results. Regarding BsmI polymorphism, the study conduct by Stefanic et al.TaqI, ApaI or BsmI polymorphism. Furthermore, subgroup analyses found similar results, and sensitivity analyses did not change the orientation of pooled results.qI rs7312 SNPs, loBsmI, ApaI and TaqI, although functionally most likely anonymous, have been associated with total and allele-specific VDR mRNA expressionet al.ApaI-BsmI LD block were not associated with Chinese patients with HT (P > 0.05). Giovinazzo et al.BsmI-TaqI haplotypes were found to be the predisposing and protective haplotypes, respectively. Similarly, common baT as well as the rare BaT haplotypes was associated with increased and decreased risk, respectively. However, we couldn’t do meta-analysis due to insufficient published data in these studies. These effects, including effects associated with rare variants or specific stimuli need further research.VDR 3′-RFLP haplotypes have been positioned within the regulatory element spanning-3′- untranslated region which contains polymorphic sequences affecting either VDR mRNA stability or VDR transcriptional activity22P = 0.0001), but VDR BsmI, ApaI, and TaqI polymorphisms were not associated with HT risk. The otherP = 0.02) while VDR TaqI, but not FokI polymorphisms is associated with HT. It is unfortunate that neither study analyzed the distributional difference of VDR polymorphisms stratified by vitamin D levels. Therefore, the mechanism and effect for the interaction of vitamin D and VDR in patients with HT need further investigations.Vitamin D, well-known for its role in calcium and bone metabolism, has important effects on immune regulation by binding to the VDR localized in T lymphocytes and macrophages4937385152535755BsmI polymorphism, which might have some impact on the final outcome. However, we used trim and fill method to assess the influence of publication bias and found that the results were not significantly changed with or without the addition of hypothetical missing studies. Heterogeneity among studies was also detected in some analyses due to ethnic difference, geographic characteristics and lifestyle. However, our sensitivity analysis showed that studies that contribute to heterogeneity did not significantly alter the conclusions of the overall OR. Third, the statistical power to detect the association may be lower because number of studies included in our meta-analysis is relatively small. However, Ioannidis et al.Several limitations should be discussed when explaining the results of our meta-analysis. First, lack of adjustments for some factors, such as age, gender, thyroid functional status, circulating vitamin D levels, or dietary vitamin D intake, which may influence the association between VDR variants and risk of HT, might bias the present results. Second, because of unpublished data or limited number of studies, significant publication bias was found in two models regarding FokI polymorphism is associated with HT risk in overall population or in Asians, but not in Caucasians. The TaqI, ApaI and BsmI polymorphisms are not associated with HT risk. Further well-designed studies with larger sample sizes and different ethnic population are needed to clarify the present findings. Furthermore, the exact causality and mechanism for the interaction of VDR and HT development need further experimental or animal mechanism studies.In conclusion, our meta-analysis suggests that the VDR ApaI”, “BsmI”, “FokI”, “TaqI” and “hashimoto’s thyroiditis”. The reference lists of identified articles and related reviews were reviewed for additional studies.We identified all the studies regarding the relationship of VDR gene polymorphisms and HT by searching PubMed, Embase, China National Knowledge Internet (CNKI), and Wan fang databases without language restrictions . The following key words and search terms were used to identify relative publications: “Vitamin D receptor”, “VDR”, “ApaI, BsmI, FokI and TaqI) and HT risk; and (3) providing the frequencies of the variants in cases and controls or providing sufficient data to calculate the estimation of odds ratios (ORs) with 95% confidence interval (95% CI). Exclusion criteria were as follows: (1) overlapping data; (2) studies without genotype frequency and genotype distribution or insufficient information for data extraction; (3) family-based study design; and (4) abstracts, reviews, comments or editorial articles lack of necessary raw data. In the case of overlapping data, only the study with the largest population was selected for this meta-analysis.Studies meeting all of the following inclusion criteria were included: (1) case-control study or cohort study; (2) investigating the association between VDR gene polymorphisms (Two investigators (XF Wang and WL Cheng) extracted data independently. Any disagreement was resolved through discussion. The extracted data included: name of the first author, year of publication, country, ethnicity, number of cases and controls, genotyping method, control sources, and genotype distribution in cases and controls.The quality of included studies was assessed by two independent reviewers (XF Wang and Y Ma) using the Newcastle-Ottawa Scale (NOS)I2 test were used to assess heterogeneity among trials. Q-test reported a P value < 0.1 or I2 > 50% was defined as significant heterogeneityP-values were two-tailed. All analyses were performed using Stata 11.0 . This article follows the PRISMA statementA random-effects model was used to incorporate within- and between-study heterogeneity as this can provide more conservative result than a fixed effects modelHow to cite this article: Wang, X. et al. Vitamin D receptor gene FokI but not TaqI, ApaI, BsmI polymorphism is associated with Hashimoto’s thyroiditis: a meta-analysis. Sci. Rep.7, 41540; doi: 10.1038/srep41540 (2017).Publisher's note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.


Miscanthus × giganteus is wildly cultivated as a potential biofuel feedstock around the world; however, the narrow genetic basis and sterile characteristics have become a limitation for its utilization. As a progenitor of M. × giganteus, M. sinensis is widely distributed around East Asia providing well abiotic stress tolerance. To enrich the M. sinensis genomic databases and resources, we sequenced and annotated the transcriptome of M. sinensis by using an Illumina HiSeq 2000 platform. Approximately 316 million high-quality trimmed reads were generated from 349 million raw reads, and a total of 114,747 unigenes were obtained after de novo assembly. Furthermore, 95,897 (83.57%) unigenes were annotated to at least one database including NR, Swiss-Prot, KEGG, COG, GO, and NT, supporting that the sequences obtained were annotated properly. Differentially expressed gene analysis indicates that drought stress 15 days could be a critical period for M. sinensis response to drought stress. The high-throughput transcriptome sequencing of M. sinensis under drought stress has greatly enriched the current genomic available resources. The comparison of DEGs under different periods of drought stress identified a wealth of candidate genes involved in drought tolerance regulatory networks, which will facilitate further genetic improvement and molecular studies of the M. sinensis. Miscanthus is a species of promising C4 perennial nonfood bioenergy grasses for cellulosic biofuel production [Miscanthus × giganteus, a hybrid generated from a cross between tetraploid Miscanthus sacchariflorus and diploid Miscanthus sinensis, has been intensively studied in Europe and North America as a biomass feedstock [M. × giganteus through breeding, posing limitations to its biomass productivity, abiotic stress tolerance, and climatic adaptation under some extreme conditions [M. × giganteus, M. sinensis was widely distributed around East Asia and it was shown that abundant wild M. sinensis resources were distributed in China providing a comparable yield and well abiotic stress tolerance in some places [The genus oduction . Specifieedstock –7. Howeveedstock , 9. Furtnditions –12. As ae places .Arabidopsis, 299 drought-inducible genes were identified through 7000 full-length cDNA microarray [Arabidopsis, 51 of them performed a similar function and revealed a considerable degree of similarity of drought stress response at the molecular level. Specially, genes involved in antioxidative metabolic pathways play an important role in detoxifying reactive oxygen species that can accumulate under drought stress conditions. In addition, it is well known that the phytohormone abscisic acid (ABA) level is essential for drought stress responses. Several genes involved in ABA biosynthesis and catabolism pathway in the drought stress responses have been identified in model plant and crops. In Arabidopsis, the 9-cis-epoxycarotenoid dioxygenase (NCED) family gene AtNCED3 transcripts are rapidly induced by drought stress, which proved that it plays a crucial role in drought stress-inducible ABA biosynthesis [Drought is a common environmental stress which induces adverse impacts on almost all aspects of plant development, growth, reproduction, and yield in a temperate area, and plants must adapt to this stress to survive , 18. Placroarray  and werecroarray . In ricecroarray . In a coynthesis . CYP707Aynthesis , 25. Sigynthesis . In consM. sinensis in Europe, a much broader range of adaptation than M. × giganteus was found in this diploid species [M. sinensis is considered possible to breed varieties with higher tolerance for frost and drought than M. × giganteus [M. sinensis are not well understood. Recently, high-throughput RNA sequencing technology was proved a powerful tool for gene discovering, gene expression, and physiological and biochemical metabolism realization under abiotic stress [M. sinensis under different drought stress stages based on transcriptome analysis, which contributes to well understand the change of regulatory mechanism processes and provide molecular bases for further revealing of metabolic networks associated with drought tolerance. The drought tolerance-related genes identified in this study aimed to provide varied candidate genes for M. sinensis genetic improvement and crop breeding.It has been well known that many wild plants show high tolerant phenotypes against abiotic stresses, such as salt, drought, and oxidative stresses –31. Base species , 32, indiganteus . Howeverc stress , 34. In M. sinensis drought tolerance genotype “M2010228” were sequenced using the Illumina HiSeq 2000 platform, and 349,393,396 raw reads were generated. After filtering and trimming the raw reads, a total of 316,200,846 high-quality clean reads were assembled into 114,747 unigenes using Trinity [50 was 1854 nt. Compared to previous de novo assembly of M. × giganteus by using ABySS and Phrap, the contigs obtaining longer than 200 bp were greater in this study, and a contig N50 length was longer than that of 1459 bp. All the unigenes were divided into two classes by gene family clustering, with a total of 65,203 distinct clusters identified which contained several similar unigene sequences (more than 70%) in each cluster and the other total of 49,544 distinct singletons generated with single unigene . Amongst them, 91,894 (80.08%) unigenes had significant hits in NT database; 84,456 (73.60%) in NR; 64,145 (55.90%) in GO; 55,807 (48.63%) in Swiss-Prot; 55,557 (48.42%) in KEGG; and 38,458 (33.52%) in COG. In total, 95,897 (83.57%) unigenes were annotated using at least one database.Unigene annotation and classification provide rich information about expression profiles and predict the potential functions of the assembly. Furthermore, various databases for annotation could shed light on intracellular metabolic pathways and biological behaviors of genes. In this study, the 114,747 obtained unigene sequences from six e value was <1e−30 (Sorghum bicolor and 28.0% to Zea mays (Zea mays L.), sorghum (Sorghum bicolor L. Moench), and sugarcane (Saccharum officinarum L.) and species in the genus Miscanthus. Domesticated and wild grass species in the Andropogoneae tribe are important sources of food, feed, fiber, and fuel [M. sinensis in genome-wide association analysis and QTL mapping [M. sinensis using single-nucleotide variant (SNV) markers which were developed by deep RNA sequencing and comparison with the genomes of sorghum maize and rice (Oryza sativa). Ma et al. [M. sinensis and revealed that sorghum has the closest phylogenetic relationship to Miscanthus by comparing the genome sequences to several grass species. Besides, the similarity of Miscanthus transcripts to the gene models and ESTs of sorghum, sugarcane, maize, rice, and Brachypodium distachyon was assessed by Barling et al. [M. sinensis RNA-seq showed that most of the gene annotations (59.5%) were aligned to the sorghum database, which is consistent with previous studies with the high utility of sorghum as a reference genome sequence for genus Miscanthus, supporting that the sequences obtained in our study were annotated properly.Within NR annotation, 74.3% of s <1e−30  and 86% s <1e−30 . For theZea mays . It is nand fuel . Previou mapping , 44–46.  mapping  construca et al.  created g et al.  and showM. sinensis transcriptome under drought stresses were very different in biological functions involving in transport and metabolism, cellular processes and signaling, and information storage and processing.The Clusters of Orthologous Groups (COGs) of proteins were delineated by comparing protein sequences encoded in complete genomes, representing major phylogenetic lineages. Each COG consists of individual proteins or groups of paralogs from at least 3 lineages and thus corresponds to an ancient conserved domain. The results of COG analysis indicated that 38,458 unigenes were annotated under 25 categories , among wM. sinensis, indicating that our transcript assemblies afforded functional annotation of a comparable percentage of gene products to that of annotated plant species despite the current lack of a reference genome sequence.As an international standardized gene functional classification system, Gene Ontology (GO) offers a dynamic updated controlled vocabulary and a strictly defined concept to describe properties of genes and their products in any organism , 51. In M. sinensis leaves when exposed to drought stress. The frequent classes in cellar component were the cell part (72.9%), cell (72.9%), organelle (62.9%), and membrane (28.5%). Under the molecular function group, binding (52.5%) and catalytic activity (49.2%) were found to be the two mainly distributed categories as described, which are in agreement with the active metabolic functions in the examined tissues. With the help of GO functional classification, a large number of the unigenes were assigned to a diverse range of experimentally derived annotation. Our annotations provide a great foundation and a valuable resource for gene expression profile analysis, gene location, and gene isolation experiment in Miscanthus species. In addition, the main GO classifications identified through de novo transcriptome analyses in fundamental biological processes, cellar component, and molecular function were similar to previous reported studies in M. sinensis, Sorghum bicolor, [Hemarthria [Miscanthus transcriptome within the Andropogoneae tribe.The distribution of all annotated unigenes in these three GO categories is shown in  binding .5% and cmarthria , suggestThe networks of gene interactions in cells could be well understood by the KEGG pathway analysis. In this study, all the unigenes were analyzed in the KEGG pathway database and 55,557 unigenes were mapped to twenty main categories including 128 different KEGG pathways . Most ofe value < 0.00001) to protein databases in the priority order of NR, Swiss-Prot, KEGG, and COG. Proteins with the highest ranks in BLAST results are taken to decide the coding region sequences of unigenes, and the coding region sequences are translated into amino sequences with the standard codon table. Unigenes that cannot be aligned to any database are scanned by EST-Scan, producing nucleotide sequence (5′→3′) direction and amino sequence of the predicted coding region. In this study, a total of 84,633 (73.8%) unigenes were predicted to be CDs among the 114,747 assembled (In order to conduct the prediction of protein coding region (CDs), all unigenes are firstly aligned by BLASTx  were detected through the transcriptome comparison of well-watered (M0) versus drought stress 5 days (M1) in genotype “M20102208.” The numbers of DEGs found in M0 versus M2 were 3467 upregulated and 4175 downregulated; in M0 versus M3, 829 upregulated and 4683 downregulated; in M0 versus M4, 4370 upregulated and 3792 downregulated; and in M0 versus M5, 7536 upregulated and 5297 downregulated . The resM. sinensis under drought stress, another group of comparison was conducted , while DEGs from M0 versus M3 have 65.0% matching to M0 versus M2, and the DEGs from M0 versus M4 have 76.9% similar to those from M0 versus M5  is a key enzyme for ABA biosynthesis, which was originally identified from maize viviparous 14 mutants playing a crucial role in drought stress-inducible genes [M. sinensis , 70% relative humidity, and 500 μmol photons m−2·s−1. After three-month establishment, all the replications were subjected to nature drought stress treatment. Prior to treatment, all the pots were well-watered and the soil water content (SWC) was measured by the Soil Moisture Equipment TDR 300 . Naturally, water stress was applied by stopping water for 30 days. Leaf samples were collected and immediately frozen in liquid nitrogen from three replications at 0- (SWC = 91.57%), 5- (SWC = 85.55%), 10- (SWC = 73.12%), 15- (SWC = 51.61%), 20- (SWC = 39.89%), and 30-day (SWC = 26.41%) drought stress treatments for RNA extraction. Total RNA was extracted using the RNeasy Plant Mini Kit  according to the manufacturer's instructions. RNA purity, concentration, and integrity were assessed using the RNA Nano 6000 Kit for the Agilent 2100 Bioanalyzer 2100 System . After RNA isolation and quality assessment, samples were stored at −80°C until the cDNA library construction and transcriptomic assay were completed.A wild drought tolerance genotype “M20102208” was collected from Sichuan Province . All the plants were propagated through rhizome division from a single individual. The plants were planted in plastic pots (20 cm in diameter and 25 cm in height) with soil mixture (50% loam with 50% fine sandy). μg of total RNA per sample was used to construct the cDNA libraries. In all, six cDNA sequencing libraries of M. sinensis were constructed using the NEB Next® Ultra™ RNA Library Prep Kit for Illumina . Initially, the total RNA was treated with RNase-free DNase I (NEB) for 30 min at 37°C and poly(A) mRNA was isolated from total RNA using poly-T oligo-linked magnetic beads. Following purification, the poly(A)-containing mRNA was fragmented into 200–250 bp pieces using fragment buffer (Ambion), and the first-strand cDNA was synthesized using random hexamer primers and the short fragments as templates. The products were then treated with RNase H, and second-strand cDNA was synthesized by DNA polymerase I (16°C for 2 h). Finally, NEBNext Adaptor with hairpin loop structure was ligated to the cDNA and the 3′ ends of the DNA fragments were adenylated in preparation for hybridization. Subsequently, the cDNA fragments were purified and the quality of the library was evaluated using the Agilent Bioanalyzer 2100 system. The index-coded samples were clustered on a cBot System using the TruSeq PE Cluster kit v3-cBot-HS (Illumina). After generating the clusters, Illumina sequencing (paired-end technology in the Illumina HiSeq 2000 platform) of the six libraries was performed for RNA-seq analysis.A total of 5 http://trinityrnaseq.github.io), which is specific for high-throughput transcript assembly of RNA-Seq data without a reference genome [e value < 1e−5). Protein function information can be predicted from the annotation of the most similar protein in those databases. If the results of the databases conflicted with each other, a priority order of NR, Swiss-Prot, KEGG, and COG was followed when deciding the sequence direction of unigenes. The KEGG pathway database records networks of molecular interactions in the cells and variants of them specific to particular organisms. GO functional annotation was conducted with NR annotation which offers a dynamic updated controlled vocabulary and a strictly defined concept to comprehensively describe properties of genes using BLAST2GO program.Raw reads (accession numbers SRP095822 in the NCBI SRA database) from six libraries were transformed from sequencing-received image data. Raw reads were filtered to remove those with only adapters, low quality reads, and unknown or reads with less than 20 bp in length. Following the calculation of sequence duplication, Q20, and the GC content of the clean reads, the de novo assembly of RNA-seq was conducted using Trinity (e genome . The unihttp://cufflinks.cbcb.umd.edu/). For genes with more than one alternative transcript, the longest transcript was selected to calculate the FPKM. With the expression level of each gene calculated, the differential expression analysis was conducted. The false discovery rate (FDR) as a statistical method was used to determine the threshold of the p value in multiple hypothesis testing, and for the analysis, a threshold of the FDR ≤ 0.001 and an absolute value of log2 ratio ≥ 1 were used to judge the significance of the gene expression differences. Tool edgeR23 was used to identify significantly up- and downregulated genes on the read count values of genes. The differentially expressed genes (DEGs) were used for GO and KEGG enrichment analyses. First, all of the DEGs were blasted in the GO database (http://www.geneontology.org/) and the gene numbers were calculated for each GO term with GO-Term Finder version 0.86 (http://search.cpan.org/dist/GO-TermFinder/). GO terms were defined as significantly enriched GO terms in DEGs, if the corrected p value was ≤0.05. Pathway enrichment analysis identifies significantly enriched metabolic pathways or signal transduction pathways in DEGs when compared with the whole genome background. Both GO terms and KEGG pathways with a Q value ≤ 0.05 are significantly enriched in DEGs.The gene expression level was calculated by the number of uniquely mapped reads per kilobase of exon fragments per million mappable reads (FPKM) using Cufflinks  unigenes were annotated to at least one database including NR, Swiss-Prot, KEGG, COG, GO, and NT, and most of the annotations (59.5%) were aligned to the sorghum database, which is consistent with previous studies with the high utility of sorghum as a reference genome sequence for genus Miscanthus, supporting that the sequences obtained in our study were annotated properly. Differentially expressed gene analysis under different stress periods indicates that drought stress 15 days (soil water content reached 51.61%) could be a critical period for M. sinensis response to drought stress. M. sinensis plays an important role in improving the genetic base, abiotic stress tolerance, and climatic adaptation for this genus as a nonfood bioenergy crop. Hence, the transcriptome sequencing of M. sinensis reported here provides useful information for gene identification and greatly enriches the genomic available resources. The comparison of DEGs under different periods of drought stress allowed us to identify a wealth of candidate genes involved in drought tolerance regulatory networks, which will facilitate further advancements in genetic and molecular mechanisms with desired traits in further M. sinensis breeding programs.High-throughput RNA sequencing technology was proved a powerful tool for gene discovering, gene expression, and physiological and biochemical metabolism realization under abiotic stress. By using the Illumina HiSeq 2000 platform to sequence


Xanthomonas oryzae pv. oryzae (Xoo), is one of the devastating diseases of rice. It is well established that the wild rice Oryza meyeriana is immune to BB. In this study, the transcriptomic analysis was carried out by RNA sequencing of O. meyeriana leaves, inoculated with Xoo to understand the transcriptional responses and interaction between the host and pathogen. Totally, 57,313 unitranscripts were de novo assembled from 58.7 Gb clean reads and 14,143 unitranscripts were identified after Xoo inoculation. The significant metabolic pathways related to the disease resistance enriched by KEGG, were revealed to plant-pathogen interaction, phytohormone signaling, ubiquitin mediated proteolysis, and phenylpropanoid biosynthesis. Further, many disease resistance genes were also identified to be differentially expressed in response to Xoo infection. Conclusively, the present study indicated that the induced innate immunity comprise the basal defence frontier of O. meyeriana against Xoo infection. And then, the resistance genes are activated. Simultaneously, the other signaling transduction pathways like phytohormones and ubiquitin mediated proteolysis may contribute to the disease defence through modulation of the disease-related genes or pathways. This could be an useful information for further investigating the molecular mechanism associated with disease resistance in O. meyeriana.Rice bacterial blight (BB), caused by  Oryza sativa L.) is one of the most important food crops in the world. However, rice production is seriously constrained by various diseasesXanthomonas oryzae pv. oryzae (Xoo) has been become a devastating disease of rice worldwide in 1960s and 1970s when the high-yielding varieties were introduced1XooRice (Xanthomonas resistance (Xa) genes specifically resistant to various strains of Xoo were identified or isolated from a variety of rice materialsXa21 is the first resistant gene derived from the wild rice (O. longistaminata)Wild rice, which is growing widely in 77 countries of the Asia, Africa, Latin America and Australia, is considered to be an important and rich gene pool6O. meyeriana Baill is a unique sourceO. meyeriana collected from different regions of South China has been demonstrated highly resistant to Xoo infection, usually accompanied by the typical hypersensitive response (HR)Xoo  level was raised after Xoo infection12O. meyeriana is regarded as a valuable gene pool. The genetic resources of O. meyeriana have been successfully used for BB resistance through asymmetric somatic hybridizationO. meyeriana and O. sativa subsp. japonicaO. meyeriana in response to the Xoo infection by the suppression subtractive hybridizationO. meyeriana under the normal growing conditionXoo infection for O. meyeriana is still limited.The awnless wild rice species of  (HR)Xoo . In contpathogen . PreviouR) genes, typically encoding the nucleotide-binding site and leucine-rich repeat (NBS-LRR) domains, to recognize these effectors and initiate a second layer of response called the effector-triggered immunity (ETI). ETI often leads to HR mediated by the R proteins in plant and is an accelerated and amplified PTI response, resulting in disease resistance to protect the plant itselfIt is well documented that plants have evolved complex innate immune systems in response to the biotic and abiotic stresses. The plant immune system is composed of surveillance systems to perceive the microbe elicitors, such as the well-characterized flagellin and peptidoglycan, which in turn allow plants to switch from the normal growth and development to a defence mode18Xoo in cultivar riceX. axonopodis pv. glycines in soybeanX. arboricola pv. pruni in peachO. meyeriana to Xoo infection, deep RNA sequencing was performed in this work. Through identifying differentially expressed genes (DEGs) of O. meyeriana after Xoo infection, the major metabolic pathways related to disease resistance including the plant innate immunity, phytohormone signaling, ubiquitin mediated proteolysis, and phenylpropanoid biosynthesis were enriched and the candidate genes targeting to the disease resistance in the plant were also inferred.Transcriptomic sequencing based on the next-generation sequencing technologies, provides a powerful and effective strategy to produce genome-scale sequence data, which allows the whole transcriptome to be investigated in a high-throughput and quantitative manner, especially for non-model organisms with genomic sequences that are yet to be measuredO. meyeriana against Xoo, total RNAs from the leaves at 0 h, 1 h, 2 h, 4 h, 24 h, and 48 h after Xoo inoculation were isolated and used for high-throughput sequencing. The RNAs were pooled into three groups: CO acts as the control; TE represents the RNAs from 1 h, 2 h and 4 h after inoculation as the early response; TO signifies the late response derived from the 24 h and 48 h RNA samples. Each group was further divided 3 sub-samples. Altogether, 9 sub-samples were applied to Illumina HiSeq2000 platform for sequencing. The sequencing data are summarized . All the contigs encoding ORFs were further assembled using the software CAP3, resulting in 57,313 unitranscripts with a mean length of 2,637 bp . All theP values ≤0.05 (Xoo infection.The relative expression level of each transcript in terms of fragment per kilobase per million (PKFM) was estimated by mapping all the clean reads from each library back to the 57,313 transcripts. Consequently, 14,143 DEUs (24.7%) were identified between the three libraries of CO, TE and TO with es ≤0.05 . The anaes ≤0.05 . ClusterTo find out the intrinsic differences in the transcriptome of the three libraries, DEUs in the libraries of TE and TO were compared with the control (CO) . TotallyO. meyeriana after Xoo infection was analyzed using GO (Gene Ontology). A total of 8,134 (57.6%) transcripts were assigned to at least one GO term. Among the other biological processes, where the term occurrences took place, metabolic process (25.6%) and cellular process (23.4%) were highly represented . The results revealed that 5,137 DEUs was assigned into 313 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways . Most ofXoo inoculation (18In the plant-pathogen interaction pathway (Ko04626), 19 DEGs containing 133 DEUs were identified as up-regulated at the early stage in response to culation . Except culation . In plan2+ concentration and activates the calcium-dependent protein kinase (CDPK) in plant cellsO. meyeriana, 4 genes involved in Ca2+ signaling were up-regulated at an early stage of Xoo infection  is an essential regulator in SA-signaling was up-regulated at the early stage in O. meyeriana. The downstream gene encoding the transcription factor TGA was showed significantly different expression at different stages, which was down-regulated between TE/CO, while up-regulated between TO/CO , jasmonic acid (JA), gibberellic acids (GAs), ethylene (ET), and brassinosteroids (BRs), act as signaling molecules to trigger or mediate a diverse array of plant immune responsesnfection . In JA sen TO/CO .Xoo infection did not induce the expression of genes encoding ethylene receptor ETR and ethylene-insensitive protein 2 (EIN2).As one of the backbone of the induced defence signaling pathway, ET signal transduction has been well documentedRecent studies indicated that the auxin and cytokinin (CK) signaling pathways may be involved in defence responseXoo infection, among them, three showed up-regulated and another one showed down-regulated  have been identified, conferring resistance to Xoo in riceO. meyeriana, two Xa genes (xa13 and Xa21) were identified as DEGs after Xoo infection  being up-regulated after Xoo infection  play important roles in various cellular processes, which contains four basal components of E1, E2, E3 and ubiquitinnfection . We beliO. meyeriana, thickening of cell wall especially in the leaf xylem tissue was reported as a defence mechanism against XooXoo infection may contribute to the cellular wall thickening of the leaf xylem tissue in O. meyeriana.The secondary phenylpropanoid metabolic pathway was revealed to be overall up-regulated. The similar results have been reported in many plant-pathogen interactionsR genes are regarded as the second mechanism of disease resistance and have been recognized in various plants. To date, about 40 R genes specific to BB have been identified in rice, and nine  have been isolated by map-based cloningXa21 and xa13 were identified as DEGs in this work, but not in previous studiesR genes that were differentially expressed in response to Xoo infection  may comprise the basal defence frontier, and then activates the R gene expression, which in turn induces HR and thickening of xylem cell wall to restrict spreading of the bacterial cells in the leaf xylem tissue of O. meyeriana.In conclusion, the transcriptomic analysis provide us with a glimpse of the transcriptional response of O. meyeriana was collected from the native niches at Yuanjiang county, Yunnan province of China, and grown in a greenhouse. For one individual plant, three leaves were inoculated with X. oryzae pv. oryzae at an inoculum concentration of 3 × 108 cells per milliliter by the leaf clipping methodXoo cell suspension, were served as the control (0 h). Three replicates of leaf samples (3 × 3 leaves) were collected for each time points.260/280 ratio ranged from 1.9 to 2.1, and OD260/230 ratio of 2.0 to 2.5, were used for the next processing.Total RNAs were extracted from all the leaf samples using Trizol reagent  according to manufacturer’s instructions. The genomic DNAs were removed by DNase I . The RNA concentration and quality were measured with the Qubit fluorometer  and the Agilent 2100 Bioanalyzer, respectively. Only the RNA samples with a RIN (RNA integrity number) >8.0, ODFor RNA sequencing, the RNA samples were combined into three groups: CO, TE and TO, which were designed for the control leaves (0 h); early stage ; and the late stage (24 h and 48 h), respectively. Each group was equally divided into 3 sub-samples. For each sub-sample, the mRNAs were further purified from the total RNAs (20 μg) using Dynaloligo (dT) 25 magnetic beads and fragmented into smaller pieces. The fragmented mRNAs were used for the first-strand cDNA synthesis with the random primer by reverse transcriptase. Second strand cDNAs were synthesized using RNase H and DNA polymerase I. The resulting cDNA fragments were further repaired at 3′-end by adding a single base (A), and then ligated with the Illumina adapters. Subsequently, the cDNA fragments were purified and enriched by PCR. The quality of cDNA libraries was measured by Agilent 2100 Bioanalyzer.http://ww.bioinformatics.bbsrc.ac.uk/projects/fastqc/). Finally, 100 bp paired-end (PE) clean reads were obtained.The cDNA libraries were sequenced using the Illumina HiSeq2000 platform. The fluorescent image processing, base-calling and calculation of quality value were performed by the Illumina data processing pipeline 1.4. Meanwhile, sequencing quality was assessed by fastQC http://www.ncbi.nlm.nih.gov/) by using local blastx program with the expect E-value cut-off <10−5.All the reassembled contigs with predicted ORFs generated by CAP3 were used for similarity search against the NR database downloaded from GenBank (http://wego.genomics.org.cn/cgi-bin/wego/index.pl). Meanwhile, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways were assigned to each contig on KEGG Automatic Annotation Server (KAAS) (http://www.genome.jp/kegg/). Thus, multiple contigs could be clustered to the same GO terms and the same KEGG pathway.The blast results were imported into the Blast2GOP-value ≤ 0.05 and the log2fold-change (log2FC) >1.In order to calculate the relative express level of each contig in different samples, all the PE reads pooled from the three sub-samples in each sample group were aligned back to the final annotated contigs by using perl scripts in the Trinity package under default parameters optionO. meyeriana leaves at 0 h, 1 h, 2 h, 4 h, 24 h and 48 h post inoculation with Xoo as described above. After treated with DNase I, first strand cDNAs were synthesized using oligo (dT) and random hexamers as primers by M-MLV reverse transcriptase  according to the manufacturer’s instructions. Real-time PCR were performed in a 25 μl volume using SYBR premix Ex TaqTM II  on the CFX Connect Real-time PCR detection system . The quantitative PCR was followed by the program: 5 min at 95 °C; followed by 45 cycles of amplification with denaturation for 5 s at 95 °C, annealing for 30 s at 55 °C, and extension for 20 s at 72 °C. Triplicate under identical conditions were synchronously performed for all selected genes. The relative expression values of each transcript were then calculated by the delta-delta Ct (2−ΔΔCt) method using the CFX Manager 3.0 of the amplifier.Twenty five DEUs were selected to verify the reliability of the RNA sequencing data by quantitative real-time PCR (RT-qPCR). The designed primers for amplification of the selected transcripts were presented in Raw sequencing data are available through the NCBI Sequence Read Archive . All samples were sequenced as 100 bp paired-end reads on an Illumina HiSeq2000 sequencer.How to cite this article: Cheng, X.-J. et al. Transcriptome analysis confers a complex disease resistance network in wild rice Oryza meyeriana against Xanthomonas oryzae pv. oryzae. Sci. Rep.6, 38215; doi: 10.1038/srep38215 (2016).Publisher’s note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.