The efficiency of molecular breeding largely depends on the cost of genotyping, which plays an important role in breeding and population genetic studies. Compared to pig, cattle, and sheep, poultry has a small size genome, and lower individual value. Developing a relatively low-density genotyping panel is needed to decrease the genotyping cost for poultries. In the present study, we developed a chicken 11 K single nucleotide polymorphism (SNP) genotyping panel for breeding and genetic analysis using genotyping by targeted sequencing (GBTS). The panel design was based on whole genome sequencing data from 7 local breeds and a commercial breed. In total 11,200 SNPs were included in the final chip. Population analyses of the eight breeds showed high SNP call rates and minor allele frequencies (MAF), enabling clear differentiation between populations. The families within population can also be identified. Additionally, the 11K panel was applied to a genomic prediction of Wenshang Barred chickens, the predictive ability and prediction accuracy between the WGS and chip dataset non-significantly differed. Therefore, the newly developed chicken low-density SNP is efficient, cost-effective, and well-suited for application in local Chinese chickens, which can accelerate chicken breeding improvements and enhance conservation efforts.

Objectives The aim of this study was to investigate the correlation between CDSS (Chinese DIC Scoring System) score and 30-day mortality in patients with intracranial hemorrhage (ICH) with nonacute promyelocytic leukemia (APL) acute leukemia. Methods This cohort study enrolled patients with non-APL acute leukemia complicated by ICH. The CDSS score was assessed in this patient population. Multivariable Cox regression was used to analyze the association between CDSS score and 30-day mortality. Additionally, interaction and stratified analyses were conducted based on variables such as age, sex, white blood cell count, platelet count, and albumin levels. Results In a study involving 82 patients diagnosed as nonAPL acute leukemia with ICH, the overall 30-day mortality rate was 61.0%, with 50 out of the 82 patients succumbing to the condition. Among those with CDSS scores ≥ 6, the mortality rate was 87% (20 out of 23), which was higher than the 50.8% mortality rate of the CDSS scores < 6 group (30 out of 59) (p = 0.002). In multivariate regression models, a 28% increase in 30-day mortality was linked to a one-point increase in CDSS score(HR = 1.28, 95% CI 1.06–1.56). Furthermore, it was associated with a 124% increase in 30-day mortality in CDSS scores ≥ 6 compared with that in the CDSS scores < 6 (HR = 2.24, 95% CI 1.1–4.56). Interaction analysis revealed no significant interactive effect on the relationship between CDSS score and 30-day mortality. Conclusions The CDSS score was linked to a higher 30-day mortality rate, particularly in patients with CDSS scores ≥ 6.

The growth curve is a vital instrument for assessing and forecasting weight and developmental shifts in livestock and poultry, which reflects the changes of bodyweight traits with time and plays a key role in guiding breeding and production approaches. This study performed a genome-wide association study (GWAS) for growth curve parameters generated by nonlinear models which fit original weight-age records, to discover the SNPs and candidate genes correlated with growth traits. Data from 362 Chinese Wenshang Barred Chickens weighed at the age of 0, 2, 4, 6, 8, 10, 12, 14, 16, and 18 weeks were used to fit the Gompertz, Logistic, and von Bertalanffy growth models. The Gompertz model showed the highest coefficient of determination (R2 = 0.974). The mature body weight (A), time scale (b), and maturity rate (k) were treated as phenotypes for single-trait GWAS. The GWAS identified 44, 103, and 5 significant single nucleotide polymorphisms (SNPs) associated with A, b, and K, respectively. Among them, several candidate genes, including LDB2, TOB2, RCBTB1, KPNA3, SLIT2, LCORL, LAP3, and TPRA1, were previously reported to be associated with growth and development. Two lead SNPs (4:76022389, 4:76070237) on the LDB2 gene were significantly associated with the growth curve. Further research of these candidate genes could help explore the full genetic architecture underlying growth and development traits in poultry.

Producing translucent eggs has been found to reduce the quality and safety of the eggs, as well as the demand from consumers. However, the intestinal function and the molecular mechanism for the production of translucent eggs remain uncertain. A total of 120 eggs from 276-day-old Jining Bairi were divided into two groups based on eggshell translucence: the translucent egg group (group T) and the normal group (group C). Group T exhibited thicker eggshells and a lower egg yolk color. Subsequently, we divided the chickens into translucent and normal groups based on their egg quality. We then assessed the plasma biochemical index, intestinal morphology and structure, enzyme activity, and antioxidant capacity of the hens producing translucent eggs compared to those producing normal eggs. The results showed that the ratio of duodenal villus length to crypt depth, succinate dehydrogenase (SDH) activity, chymotrypsin, total ATPase (T-ATPase), alkaline phosphatase (AKP), and glutathione peroxidase (GSH-Px) activities were decreased in the hens produced translucent eggs (p < 0.05), but malondialdehyde (MDA) content was increased (p < 0.05); jejunal lipase activity, Na+K+-ATPase activity, total antioxidant capacity (T-AOC), and GSH-Px activities were decreased (p < 0.05) in group T; ileal amylase and Ca2+Mg2+-ATPase activities were also decreased (p < 0.05) in group T. In addition, we identified a total of 471 differentially expressed genes (DEGs) in duodenal tissue, with 327 up-regulated genes and 144 down-regulated genes (|log2FC| ≥ 1 and p < 0.05). Enrichment analysis showed that the up-regulated genes, such as GSTT1, GSTO2, and GSTA3, were mostly enriched in metabolism of xenobiotics by cytochrome P450, drug metabolism-cytochrome P450, and oxidative phosphorylation pathways. The results of our study indicate that plasma lipid metabolism disorder, decreased intestinal antioxidant capacity, and altered intestinal metabolism capabilities may influence the formation of translucent eggs.

To study the therapeutic effect of sinomenine hydrochloride (SH) on dextran sodium sulfate (DSS)-induced colitis in mice as an animal model and the changes of Notch signaling pathway in colon tissue of mice after treatment. Twenty-four mice were randomly divided into control group, model group, SH low-dose group (20 mg/kg) and SH high-dose group (60 mg/kg), with 6 mice in each group. Disease activity index (DAI), colonic mucosal injury index and colonic histopathological score were calculated. The expression levels of related genes, proteins in Notch signaling pathway and inflammatory factors were quantified. SH can significantly reduce the symptoms of colitis mice, and can significantly reduce the DAI score (Model: 3.44 ± 0.27; SH-20: 2.50 ± 0.18; SH-60: 1.89 ± 0.17; P < 0.001) and histopathological injury degree (Model: 7.67 ± 0.52; SH-20: 5.17 ± 0.75, P < 0.01; SH-60: 3.33 ± 0.52, P < 0.001). SH can down-regulate the expression levels of Notch1, NICD1, Jagged1 and Hes1 proteins in colon tissue of colitis mice (Model: 1.92 ± 0.16, 1.83 ± 0.21, 2.23 ± 0.22, 1.91 ± 0.17; SH-20: 1.56 ± 0.12, 1.39 ± 0.13, 1.58 ± 0.12, 1.38 ± 0.11; SH-60: 1.24 ± 0.09, 1.23 ± 0.10, 1.23 ± 0.11, 1.22 ± 0.09; P < 0.01), and reduce the contents of serum pro-inflammatory cytokines TNF-α, IL-1β and IL-6 (Model: 718.53 ± 81.81, 51.62 ± 2.80, 444.07 ± 67.77; SH-20: 544.72 ± 90.03, 34.10 ± 2.90, 345.43 ± 43.40; SH-60: 434.11 ± 71.75, 29.44 ± 3.70, 236.11 ± 29.35; P < 0.001). The therapeutic effect of SH on DSS-induced colitis in mice may be related to inhibiting the overactivation of Notch signaling pathway.

Many different chicken breeds are found around the world, their features vary among them, and they are valuable resources. Currently, there is a huge lack of knowledge of the genetic determinants responsible for phenotypic and biochemical properties of these breeds of chickens. Understanding the underlying genetic mechanisms that explain across-breed variation can help breeders develop improved chicken breeds. The whole-genomes of 140 chickens from 7 Shandong native breeds and 20 introduced recessive white chickens from China were re-sequenced. Comparative population genomics based on autosomal single nucleotide polymorphisms（SNPs） revealed geographically based clusters among the chickens. Through genome-wide scans for selective sweeps, we identified thyroid stimulating hormone receptor（TSHR, reproductive traits, circadian rhythm）, erythrocyte membrane protein band 4.1 like 1（EPB41L1,body size）, and alkylglycerol monooxygenase（AGMO, aggressive behavior）, as major candidate breed-specific determining genes in chickens. In addition, we used a machine learning classification model to predict chicken breeds based on the SNPs significantly associated with recourse characteristics, and the prediction accuracy was 92%, which can effectively achieve the breed identification of Laiwu Black chickens. We provide the first comprehensive genomic data of the Shandong indigenous chickens. Our analyses revealed phylogeographic patterns among the Shandong indigenous chickens and candidate genes that potentially contribute to breed-specific traits of the chickens. In addition,we developed a machine learning-based prediction model using SNP data to identify chicken breeds. The genetic basis of indigenous chicken breeds revealed in this study is useful to better understand the mechanisms underlying the resource characteristics of chicken.

Background Metabolic derangements and systemic inflammation are related to the progression of colorectal cancer (CRC) and the prognoses of these patients. The survival of stage II and III CRC patients existed considerable heterogeneity highlighting the urgent need for new prediction models. This study aimed to develop and validate prognostic nomograms based on preoperative serum liver enzyme as well as evaluate the clinical utility. Methods A total of 4014 stage II/III primary CRC patients pathologically diagnosed from January 2007 to December 2013 were included in this study. These patients were randomly divided into a training set ( n = 2409) and a testing set ( n = 1605). Univariate and multivariate Cox analyses were used to select the independent factors for predicting overall survival (OS) and disease-free survival (DFS) of stage II/III CRC patients. Next, nomograms were constructed and validated to predict the OS and DFS of individual CRC patients. The clinical utility of nomograms, tumor-node-metastasis (TNM), and the American Joint Committee on Cancer (AJCC) system was evaluated using time-dependent ROC and decision curve analyses. Results Among seven preoperative serum liver enzyme markers, aspartate aminotransferase-to-alanine aminotransferase ratio (De Ritis ratio) was identified as an independent factor for predicting both OS and DFS of stage II/III CRC patients. The nomograms incorporated De Ritis ratio and significant clinicopathological features achieved good accuracy in terms of OS and DFS prediction, with C-index of 0.715 and 0.692, respectively. The calibration curve showed good agreement between prediction by nomogram and actual observation. The results of time-dependent ROC and decision curve analyses suggested that the nomograms had improved discrimination and greater clinical benefits compared with TNM and AJCC staging. Conclusions De Ritis ratio was an independent predictor in predicting both the OS and DFS of patients with stage II/III CRC. Nomograms based on De Ritis ratio and clinicopathological features showed better clinical utility, which is expected to help clinicians develop appropriate individual treatment strategies for patients with stage II /III CRC.

Supplemental tables S1-5 and figures S1-2. Supplemental Tables including: (1)Supplementary Table S1. MOOSE Checklist; (2)Supplementary Table S2. Study Quality Scores of Prospective Cohort Studies Using Newcastle- Ottawa Scale; (3)Supplementary Table S3. Additional Information of Included Studies; (4)Supplementary Table S4. The Dose-Response analysis of TC, HDL, and LDL; and (5)Supplementary Table S5. Sensitivity analysis. Supplemental Figures including: (1)Supplementary Fig. S1. Selection of Studies for Inclusion in Meta-analysis; and (2)Supplementary Fig. S2. Funnel Plots of Publication Bias for Total Cholesterol.

In this study, we explored the characteristics and causes of freckle formation. We collected 15 normal and freckled eggs each for eggshell index testing and hypothesized that the structure and function of the uterus would have a direct effect on freckled egg production given that eggshells are formed in the uterus. To test this hypothesis, we collected uterine tissue from laying hens (418 days of age) that laid normal (Group C, n = 13) and freckled (Group T, n = 16) eggs for 7 consecutive days. When we examined the eggshell quality, we found that the L value was significantly lower (P < 0.05) in the freckled site group of freckled eggs compared to the normal egg group during the detection of blunt pole, equator, and sharp pole of the eggshell color. The a-values of the three positions were significantly higher (P < 0.05) in the freckled site group of freckled eggs, and the a-values of the blunt pole were significantly lower (P < 0.05) in the background site group of freckled eggs, compared to the normal egg group. The b-values were significantly higher (P < 0.05) at three locations in the freckled site group of freckled eggs compared to the normal egg group. During the detection of eggshell thickness, the blunt pole was significantly higher (P < 0.05) in the freckled egg site group of freckled eggs compared to the normal egg group, and there was no significant difference between the other groups (P > 0.05). There was no significant difference (P > 0.05) between the transverse and longitudinal diameters of the eggs in each group.We then performed histopathology and transcriptome analyses on the collected tissue. When compared with group C, uterine junctional epithelial cells in group T showed significant defects and cilia loss, and epithelial tissue was poorly intact. From transcriptomics, genes that met (|log2FC|) ≥ 1 and P < 0.05 criteria were screened as differentially expressed genes (DEGs). We identified a total of 136 DEGs, with 101 up- and 35 down-regulated genes from our RNA-seq data. DEGs identified by enrichment analyses, which were potentially associated with freckled egg production were: IFI6, CCL19, AvBD10, AvBD11, S100A12, POMC, and UCN3. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses showed that pathways were associated with immunoreaction and stress stimulation, e.g., complement activation, interleukin-1 cell reactions, viral responses, cell reactions stimulated by corticotropin releasing hormone, steroid hormone mediated signaling pathways, staphylococcal infections, B cell receptor signaling pathways, and natural killer cell mediated cytotoxicity. From these data, freckled areas deepen freckled eggshell color, but background areas are not affected. At the same time,we reasoned that freckle eggs may result from abnormal immune responses and impaired uterine functions induced by stress. Therefore, the uterus of laying hens in a state of stress and abnormal immune function can cause the appearance of freckled eggs.

This study investigated the effects of ferulic acid (FA) on the oxidative stability and nitrozation of porcine myofibrillar proteins (MPs) under oxidative stress. The addition of FA significantly (P < 0.05) inhibited the formation of carbonyl and dityrosine, and reduced the sulfhydryl and free amine content compared with the oxidized MPs. Moreover, FA intensified oxidation-initiated loss of surface hydrophobicity and α-helix conformation, and enhanced protein cross-linking. A progressive quenching of MP endogenous fluorescence intensity was observed with the increase of FA concentration, and the emission wavelength showed a slight red shift. Beside, the addition of FA significantly eliminated sodium nitrite and almost completely inhibited the formation of 3-nitrotyrosine (3-NT) and N-nitrosodimethylamine (NDMA) at 200 μmol/g. Principal component analysis indicated that there were positive correlations between protein oxidation variables (carbonyl, dityrosine and surface hydrophobicity) and nitrozation indexes (3-NT and NDMA), as well as between 3-NT and NDMA. These results suggest that FA could suppress the nitration via controlling MP's oxidation.

Background Nonpharmacologic interventions that modify lifestyle can lower blood pressure (BP) and have been assessed in numerous randomized controlled trials and pairwise meta‐analyses. It is still unclear which intervention would be most efficacious. Methods and Results Bayesian network meta‐analyses were performed to estimate the comparative effectiveness of different interventions for lowering BP. From 60 166 potentially relevant articles, 120 eligible articles (14 923 participants) with a median follow‐up of 12 weeks, assessing 22 nonpharmacologic interventions, were included. According to the surface under the cumulative ranking probabilities and Grading of Recommendations Assessment, Development and Evaluation (GRADE) quality of evidence, for adults with prehypertension to established hypertension, high‐quality evidence indicated that the Dietary Approach to Stop Hypertension (DASH) was superior to usual care and all other nonpharmacologic interventions in lowering systolic BP (weighted mean difference, 6.97 mm Hg; 95% credible interval, 4.50–9.47) and diastolic BP (weighted mean difference, 3.54 mm Hg; 95% credible interval, 1.80–5.28). Compared with usual care, moderate‐ to high‐quality evidence indicated that aerobic exercise, isometric training, low‐sodium and high‐potassium salt, comprehensive lifestyle modification, breathing‐control, and meditation could lower systolic BP and diastolic BP. For patients with hypertension, moderate‐ to high‐quality evidence suggested that the interventions listed (except comprehensive lifestyle modification) were associated with greater systolic BP and diastolic BP reduction than usual care; salt restriction was also effective in lowering both systolic BP and diastolic BP. Among overweight and obese participants, low‐calorie diet and low‐calorie diet plus exercise could lower more BP than exercise. Conclusions DASH might be the most effective intervention in lowering BP for adults with prehypertension to established hypertension. Aerobic exercise, isometric training, low‐sodium and high‐potassium salt, comprehensive lifestyle modification, salt restriction, breathing‐control, meditation and low‐calorie diet also have obvious effects on BP reduction.

Background Differential DNA methylation panel derived from peripheral blood could serve as biomarkers of CRC susceptibility. However, most of the previous studies utilized post-diagnostic blood DNA which may be markers of disease rather than susceptibility. In addition, only a few studies have evaluated the predictive potential of differential DNA methylation in CRC in a prospective cohort and on a genome-wide basis. The aim of this study was to identify a potential panel of DNA methylation biomarkers in peripheral blood that is associated with CRC risk and therefore serve as epigenetic biomarkers of disease susceptibility. Methods DNA methylation profile of a nested case-control study with 166 CRC and 424 healthy normal subjects were obtained from the Gene Expression Omnibus (GEO) database. The differentially methylated markers were identified by moderated t-statistics. The DNA methylation panel was constructed by stepwise logistic regression and the least absolute shrinkage and selection operator in the training dataset. A methylation risk score (MRS) model was constructed and the association between MRS and CRC risk assessed. Results We identified 48 differentially methylated CpGs sites, of which 33 were hypomethylated. Of these, sixteen-CpG based MRS that was associated with CRC risk (OR = 2.68, 95% CI: 2.13, 3.38, P < 0.0001) was constructed. This association is confirmed in the testing dataset (OR = 2.02, 95% CI: 1.48, 2.74, P < 0.0001) and persisted in both males and females, younger and older subjects, short and long time-to-diagnosis. The MRS also predicted CRC with AUC 0.82 (95% CI: 0.76, 0.88), indicating high accuracy. Conclusions Our study has identified a novel DNA methylation panel that is associated with CRC and could, if validated be useful for the prediction of CRC risk in the future.

黄酮类化合物是植物源食品中最重要的一类活性成分,具有抗炎、抗癌、预防心血管疾病等生物学功能.自噬通过降解细胞内受损的线粒体、折叠的蛋白体、病毒等物质维持细胞内的稳态.它分为巨自噬、微自噬和分子伴侣介导的自噬,其中巨自噬是主要类型.黄酮类物质能够通过不同的信号通路调控自噬进而干预疾病,其中磷脂酰肌醇3激酶/蛋白激酶B/雷帕霉素的哺乳动物靶标(Phosphatidylinosittide 3-kinase/Protein kinaseB/mammalian target of Rapamycin,PI3 K/AKT/mTOR)是主要的调控自噬的信号通路.本文针对植物源食品中几种常见的黄酮类化合物,综述了其通过自噬干预炎症、癌症、肝病以及心血管疾病等发生的分子机制和研究进展.

Abstract Objective The significance of gene methylation in peripheral blood leukocytes (PBLs) for assessing cancer prognosis is poorly understood. Our purpose is to assess the association between PBX3 methylation in PBLs and colorectal cancer (CRC) prognosis. Methods A total of 369 CRC patients were followed up for up to 10 years in this cohort study. PBL PBX3 methylation levels were determined by methylation‐sensitive high‐resolution melting. Cox regression models and Log‐rank tests were used to analyze the associations between PBX3 methylation status and CRC prognosis with a propensity score (PS) method to control confounding biases. Results In this study, we found that CRC patients with PBL PBX3 hypermethylation status had a better overall survival (OS) (hazard ratio [HRPS‐adjusted], 0.72 [95% CI, 0.52‐1.00]; P = 0.049). Subgroup analyses showed that the beneficial effect of PBX3 hypermethylation status on CRC 10‐years OS remained significant among UICC stage III patients ([HRPS‐adjusted], 0.60 [95% CI, 0.38 to 0.95]; P = 0.029) and colon cancer patients ([HRPS‐adjusted], 0.49 [95% CI, 0.26 to 0.92]; P = 0.027). Conclusion PBL PBX3 hypermethylation is positively associated with better prognosis of CRC, especially for the UICC stage III CRC patients and colon cancer patients.

Iron-refractory iron deficiency anaemia (IRIDA), a rare autosomal recessive genetic disorder caused by TMPRSS6 mutations (Finberg et al, 2008), is characterized by hypochromic microcytic anaemia, low serum iron (SI) and transferrin saturation (TS), and normal or low serum ferritin (SF). Due to impaired inhibition of hepcidin gene (HAMP) transcription by TMPRSS6 mutations (Du et al, 2008), IRIDA patients exhibit inappropriately elevated levels of hepcidin, are refractory to oral iron and only partially responsive to parenteral iron administration (De Falco et al, 2013). The optimal alternative therapy still remains uncertain when the patient is unresponsive to intravenous iron and erythropoietin. To date, 32 IRIDA families with 50 patients of different ethnic origin have been reported, accounting for 40 different mutations in the TMPRSS6 gene (De Falco et al, 2013). Several common single nucleotide polymorphisms (SNPs), including the non-synonymous polymorphisms V736A, K253E, G228D, R446W, V795I and K225E, have been significantly associated with haemoglobin (Hb) levels, iron status and erythrocyte mean corpuscular volume (MCV) in the general population (Chambers et al, 2009; Beutler et al, 2010). Sole SNPs in TMPRSS6 are not likely to be associated with IRIDA. However, SNPs may contribute to IRIDA when carried in trans with a sporadic TMPRSS6 mutation (Beutler et al, 2010). A 10-year-old girl, who presented with hypochromic microcytic anaemia at 15 months old, was referred to our hospital in December 2012. She had been diagnosed with nutritional iron deficiency anaemia in a local hospital and treated with oral iron therapy (ferrous succinate 150 mg/d) combined with ascorbic acid supplement (30 mg/d) for 4 months but with little response. She received three red blood cell transfusions (total, three units), with the last transfusion being 13 months earlier. After admission to our hospital, laboratory evidence showed severe microcytic, hypochromic anaemia [Hb, 58 g/l; MCV, 61·8 fl; mean corpuscular haemoglobin (MCH), 17·2 pg], low SI (2·04 μmol/l) and TS (7·0%) and normal SF (143 μg/l). Ultrasonic examination disclosed splenomegaly (14·8 × 4·0 cm). Her parents were healthy with normal blood counts, MCV, MCH, SI, TS and SF. Gastrointestinal defects and blood loss as causes of iron deficiency were ruled out by specific tests. Thalassaemia, sideroblastic anaemia and the common forms of congenital dyserythropoietic anaemia were also excluded. To confirm the diagnosis of IRIDA, serum hepcidin was measured by enzyme-linked immunosorbent assay (DRG Diagnostics GmbH, Marburg, Germany) and TMPRSS6 mutations were screened by DNA sequencing. The study was approved by our institutional review board and written informed consent for genetic analysis and collection of clinical and biochemical parameters were obtained from the patient's guardians according to the tenets of the Declaration of Helsinki. We detected an inappropriately high level of hepcidin (11·96 nmol/l) in the patient, which was above the normal range (3·95 ± 1·54 nmol/l) for females in our laboratory. Exon sequencing of TMPRSS6 revealed unique tri-allelic mutations in exon 7 and 17: homozygous c.757 A>G (K253E) and heterozygous c.2207 T>C (V736A) in the patient with IRIDA (Fig 1). Her mother was haploid bi-allelic heterozygous for K253E and V736A and her father was heterozygous for K253E. V736A is the most common non-synonymous polymorphism and was reported to have significant effects on both iron status (SI and TS) and erythropoiesis (Hb and MCV) in the general population via partial modulation of hepcidin transcription (Chambers et al, 2009). However, the V736A variant has not been associated with IRIDA, and none of the 50 previous cases were reported to carry this non-synonymous polymorphism (De Falco et al, 2013). K253E is also a common non-synonymous polymorphism; a previously reported Japanese female with IRIDA who was homozygous for K253E had mild anaemia (108 g/l) and responded to intravenous iron (131 g/l) (Sato et al, 2011). Here, we identified for the first time, a novel tri-allelic non-synonymous polymorphism resulting in the IRIDA phenotype with a severe hypochromic microcytic anaemia (58 g/l) and no response to oral and intravenous iron therapy in a Chinese girl. Within the first 6 weeks of hospitalization, the patient was administered with intravenous iron (75 mg/week) in combination with recombinant human erythropoietin (rhEPO, 125 μ/kg per week) but with no response. Whether rhEPO therapy is able to improve anaemia in IRIDA remains debatable. To our knowledge, only three cases with IRIDA receiving EPO have been reported so far. One of a pair of twins heterozygous for P686 fs and A118D achieved a satisfactory improvement in their anaemia after treatment with combination of intravenous iron and rhEPO (Ramsay et al, 2009). In contrast, the Hb level of two Turkish patients with K636AfsX17 did not increase after administration of rhEPO alone (Lehmberg et al, 2013). Therefore, we conducted an innovative attempt to improve anaemia by using glucocorticoid alone (i.v. methylprednisolone 0·8 mg/kg per d, for 4 and 6 weeks, respectively). Unexpectedly, during the first course of methylprednisolone, we observed a significant increase in the Hb level, from 58 g/l to 89 g/l after 2 weeks, reaching a maximum of 124 g/l in 1 month (Fig 2). More importantly, the hepcidin levels simultaneously decreased, from 11·96 nmol/l to within the normal range (3·57 nmol/l at 2 weeks; 2·66 nmol/l at 4 weeks). SF also dropped (from 143 μg/l to 104 μg/l), but there were no improvements in SI, TS, MCV and MCH when compared to baseline values. After cessation of methylprednisolone, hepcidin levels sharply increased and reached a peak value of 19·55 nmol/l within 2 months, accompanied by a decrease in the Hb level to 69 g/l. The second course of methylprednisolone was administered and the same results were repeatedly validated. These data suggested that glucocorticoids could ameliorate the IRIDA phenotype by negatively regulating hepcidin levels, but the exact molecular targets remained elusive. Collectively, these results extended the pattern of TMPRSS6 mutations and provided a promising therapeutic approach in IRIDA. This work was supported by PUMC Youth Fund to JS and the Fundamental Research Funds for the Central Universities of China to YZ (no. 2012N05). The authors thank all of the doctors and nurses in Therapeutic Centre of Anaemic Diseases and the researcher team of Clinical Laboratory Centre for their professional assistance. N.N. and J.S. designed the research and wrote the paper. N.N., J.S. and M.G. performed the research and analysed the data. Y.S., X.L., J.H., J.Z., Z.H. and D.L. contributed to the clinical data collection and sample preparation. J.S. and Y.Z. revised the manuscript. All authors made significant contributions to, reviewed, and approved the final version of the manuscript. The authors declare no conflicts of interest.

AIM OF THE STUDY:This study investigated the pattern, predictors, and recurrence of node metastasis in papillary thyroid cancer patients.MATERIAL AND METHODS:One hundred and 65 papillary thyroid cancer (PTC) patients who underwent total thyroidectomy and cervical lymph node (LN) dissection (LND), in which more than 12 lymph nodes were dissected, were examined. The nodes were classified from levels I to VI. Final pathologic diagnosis of positive lymph node metastases in the differential node levels was determined.RESULTS:Cervical metastases of PTC were most commonly encountered in level VI, followed by levels III and IV, and then levels II and V. Metastases in level I seldom occurred. Skip metastases occurred in nine patients. Univariate analysis suggested that multifocality and extracapsular invasion were associated with LN metastases. The metastatic ratio for micro PTC and local canceration derived from benign lesions and encapsulation was low. Multivariate analysis showed that LN metastases were closely related to invasion of the thyroid capsule and primary PTC. Standardized estimation showed that the encapsulating pattern had the greatest impact on developing cervical LN metastases. Lymph node recurrence was observed in 11 patients.CONCLUSIONS:The metastatic pattern of PTC assists in delineating the extent of selective LND. Routine bilateral central node dissection at the time of thyroidectomy is recommended. Comprehensive selective LND is recommended in multifocal PTC and with capsular invasion.

Objective To specifically recognized the clinical features of Aeromonas septicaemia in the patients with acute leukemia,and to provide a basis for estimates of clinical conditions and treatment options.Methods From 2002 to 2009,all patients suffered from Aeromonas septicaemia with hematologic diseases in our hospital(in northern of China) were included for the retrospective study.We analysed the demographic,clinical and microbiologic characteristics,treatment and prognosis in these patients.Results During the 8-year period,16 patients with acute leukemia developed Aeromonas septicaemia.The majority of them were males(12∶4).All of the infection was nosocomial,and Aeromonas septicaemia generally occurred in summer or fall(87.5%).No cluster of Aeromonas septicaemia was found during the study period.The isolates speciated comprised Aeromonas sobria and A.hydrophila,and A.sobria was the most common species.six(24.4%) episodes of septicaemia were polymicrobial,and in the patients of A.sobria septicaemia.All of patients recent had received or underwent antineoplastic chemotherapy,and were in severe neutropenia at the onset of septicaemia.The main clinical manifestations were fever and chill(100%),followed by diarrhea(31.3%),septic shock(25%),and so on.The most severa secondary infections were skin soft-tissue infections/cellulitis(37.5%),anorectal infections(31.3%) and pneumonia(25.0%).Most of β lactam agents(excluding penicillins) had high activity against the isolated strains.All patients received timely treatment of sensitive antibiotics.However,five(31.3%) patients died within 3 to 4 days of onset of septicaemia.A.sobria polymicrobial septicaemia complicated by septic shock and accompanied by other severe infections was the high risk factor for mortality.Conclusion Aeromonas septicaemia is uncommon in the northern of China but can occur in summer or fall,and develop particularly in the patients with acute leukemia.Chemotherapy and neutropenia were the preceding host conditions.Most of β lactam agents(excluding penicillins) had high activity against the isolated Aeromonas.A.sobria was suggested to be more pathogenic.In patients with A.sobria polymicrobial septicaemia,septic shock and secondary infections should be prevented and treated early.

Withaferin A (WA), a naturally occurring steroidal lactone, directly binds to Hsp90 and leads to the degradation of Hsp90 client protein. The purpose of this study is to investigate the structure activity relationship (SAR) of withanolides for their inhibition of Hsp90 and anti-proliferative activities in pancreatic cancer cells. In pancreatic cancer Panc-1 cells, withaferin A (WA) and its four analogues withanolide E (WE), 4-hydroxywithanolide E (HWE), 3-aziridinylwithaferin A (AzWA) inhibited cell proliferation with IC50 ranged from 1.0 to 2.8 μM. WA, WE, HWE, and AzWA also induced caspase-3 activity by 21-, 6-, 11- and 15-fold, respectively, in Panc-1 cells, while withaperuvin (WP) did not show any activity. Our data showed that WA, WE, HWE, and AzWA, but not WP, all directly bound to Hsp90 and induced Hsp90 aggregation,hence inhibited Hsp90 chaperone activity to induce degradation of Hsp90 client proteins Akt and Cdk4 through proteasome-dependent pathway in pancreatic cancer cells. However, only WA, HWE and AzWA disrupted Hsp90-Cdc37 complexes but not WE and WP. SAR study suggested that the C-5(6)-epoxy functional group contributes considerably for withanolide to bind to Hsp90, inhibit Hsp90 chaperone activity, and result in Hsp90 client protein depletion. Meanwhile, the hydroxyl group at C-4 of ring A may enhance withanolide to inhibit Hsp90 activity and disrupt Hsp90-Cdc37 interaction. These SAR data provide possible mechanisms of anti-proliferative action of withanolides.

Objective:To clarify the clinical and experimental characteristics of erythrocyte pyrimidine 5'-nucleotidase(P5'N)deficiency.Method:Nine patients with P5'N deficiency were retrospectively analyzed and related literatures were reviewed.Result:All of the patients had anemia.The concomitant symptoms including splenomegaly in 7 patients,jaundice in 6 patients,cholelithiasis in 2 patients and hepatomegaly in 1 patient.Two patients were complicated with β-thalassaemia trait.One patient was associated with pyruvate kinase(PK)deficiency.In 2 of 4 assessable patients,P5'N deficiency in first-degree relatives was detected.Four patients needed blood transfusions.One patient was splenectomised.Conclusion:The hereditary erythrocyte P5'N deficiency's major characteristic is the erythrocyte P5'N deficiency and marked red cell basophilic stippling.Acquired P5'N deficiency had been well described in associated with β-thalassaemia trait and other diseases.

To evaluate the efficacy and safety of interferon-alpha-2b (IFN-α-2b) in polycythemia vera patients(PV patient) with or without post-polycythemic myelofibrosis (post-PV MF), 30 patients with mutated JAK2V617F were enrolled in this study, from which 29 patients were evaluable. The percentage of mutated JAK2V617F allele (V617F%) was evaluated by real-time polymerase chain reaction (RT-PCR) before and after treatment with IFN-α-2b. The correlation of V617F allele burden with the major clinical outcomes was studied. Adverse effects appeared in patients was observed. The results showed that the median follow-up was 24 (12 - 42) months for 29 evaluable patients. Complete hematologic response was achieved in 10%, 48%, 72% and 78% of patients after treatment for 6, 12, 24 and 36 months respectively. The detection of V617F allele burden revealed that the molecular remission of patients (V617F%) was achieved in 41%, 76%, 89% and 89% after treatment for 6, 12, 24 and 36 months respectively. Molecular complete remission (JAK2V617F undetectable) was achieved in 4 patients, lasted from 6 to 12 months after IFN-α-2b discontinuation. The decrease of V617F% in patients with post-PV MF was significantly higher than that in patients without post-PV MF (53 ± 18% vs 32 ± 22%, respectively; p = 0.031) after treatment for 12 months. PV patients had a good tolerance to IFN-α-2b. It is concluded that IFN-α-2b can decrease the mutated V617F allele burden. Patients with PV, especially with post-PV MF, can achieve molecular remission after treatment with IFN-α-2b.