[The association between portal vein thrombosis and rebleeding after non-urgent endoscopic treatment of esophagogastric varices].

Objective: To investigate the association between portal vein thrombosis and rebleeding after non-urgent endoscopic treatment of esophagogastric varices. Methods: The cirrhotic patients with esophagogastric varices diagnosed in the People's Hospital of Zhengzhou University from January 2017 to March 2023 were retrospectively collected. The patients were divided into thrombotic group and non-thrombotic group according to the presence or absence of portal vein thrombosis. The failure rate of endoscopic treatment and rebleeding rate in different periods were compared between the two groups. Receiver operating characteristic (ROC) curve was used to select the best cutoff value of gastric varicose diameter that affected total rebleeding during follow-up in both groups. The influencing factors of rebleeding within 12 and 36 months in both groups were analyzed, and the influencing factors of rebleeding within 36 months in thrombus group were further analyzed. Results: A total of 106 patients were enrolled, including 53 patients in the thrombotic group [male 37, female 16, aged 18-78 (54±13) years] and 53 patients in the non-thrombotic group [male 37, female 16, aged 27-83 (55±12) years]. The follow-up time of the two groups were (20±15) and (25±15) months, respectively. The total rebleeding rate in the thrombotic group was higher than that in the non-thrombotic group [30.2% (16/53) vs 13.2% (7/53), P˂0.05]. The rebleeding rates within 6, 12, 24 and 36 months in the thrombotic group were higher than those in the non-thrombotic group [18.9% (10/53) vs 5.7% (3/53), 18.9% (10/53) vs 5.7% (3/53), 28.3% (15/53) vs 9.4% (5/53), 30.2% (16/53) vs 11.3% (6/53), all P˂0.05]. The best cut-off value of the diameter of gastric varices that affects the total rebleeding in the two groups was 10.4 mm (10 mm was selected as the best cut-off value for the convenience of practical clinical application). Hemoglobin ˂ 85 g/L (HR=0.202, 95%CI: 0.043-0.953, P=0.043), 10 mm ˂ the diameter of GV ≤ 15 mm (HR=5.321, 95%CI: 1.161-24.390, P=0.031) and endoscopic variceal ligation combined with endoscopic tissue adhesive injection (EVL+ETAI) (HR=7.172, 95%CI: 1.910-26.930, P=0.004) were the risk factors for the first gastroesophageal variceal rebleeding within 12 months after non-urgent endoscopic treatment. EVL+ETAI (HR=3.811, 95%CI: 1.441-10.084, P=0.007) and portal vein thrombosis (HR=4.026, 95%CI: 1.483-10.932, P=0.006) were the risk factors for the first gastroesophageal variceal rebleeding within 36 months after non-urgent endoscopic treatment. The study found that, 10 mm ˂ the diameter of GV ≤ 15 mm (HR=7.503, 95%CI: 1.568-35.890, P=0.012) was the risk factor for rebleeding within 36 months in the thrombotic group. Conclusion: Portal vein thrombosis is a risk factor for rebleeding after non-urgent endoscopic treatment of esophagogastric varices.

[Risk factors and nutritional status analysis in patients with liver cirrhosis and concomitant chronic periodontitis].

Objective: To study and explore the prevalence, characteristics, preliminary risk factors, as well as their relationship with nutritional scores in liver cirrhotic patient with chronic periodontitis. Methods: 163 patients with liver cirrhosis who were hospitalized in the Hepatology Division, Department of Internal Medicine at Tianjin Third Central Hospital from June to September 2018 were enrolled as the case group, while the control group consisted 140 healthy individuals enrolled during the same period. Periodontal examination, biochemical examination and oral hygiene habits were investigated. The prevalence of periodontitis in the two groups was compared, and the risk factors of severe periodontitis were conducted by multivariate regression analysis. Results: The prevalence of chronic periodontitis was significantly higher in patients with liver cirrhosis than healthy control population, and the differences were statistically significant (P < 0.05). The prevalence of severe periodontitis and full edentulous jaws was significantly higher in patients with liver cirrhosis than healthy control group, and the differences were statistically significant (P < 0.05 and P < 0.001). Compared with the healthy control group, the depth of periodontal pocket and the degree of attachment loss were significantly increased in the liver cirrhosis group (P < 0.001). Multivariate regression analysis showed that liver cirrhosis was the independent risk factors for both groups of patients with severe periodontitis (χ (2) = 11.046, P < 0.001). Univariate and multivariate regression analysis showed that toothbrushing frequency, nutritional risk score, prealbumin level and Child-Pugh grade were independent risk factors for occurrence of severe periodontitis in liver cirrhotic patient (χ (2) = 5.252, P = 0.022; χ (2) = 24.162, P < 0.001; χ (2) = 4.159, P = 0.041; χ (2) = 9.249, P = 0.002). Conclusion: The prevalence of periodontitis is significantly higher in patients with liver cirrhosis than healthy individuals, and liver cirrhosis is an independent risk factor for the occurrence of severe periodontitis. Toothbrushing frequency, nutritional risk score, prealbumin level and Child-Pugh grade are risk factors for severe periodontitis in patients with liver cirrhosis.

[Role of 1, 25-dihydroxyvitamin D3 in liver lipid metabolism induced by methionine-choline-deficient diet in rats].

Objective: To investigate the role of 1, 25-dihydroxyvitamin D3 [1.25(OH) (2)D(3)] in liver lipid metabolism so as to provide the clues for elucidating the mechanism of non-alcoholic fatty liver. Methods: 26 SD rats were randomly divided into control group (methionine-choline-sufficient diet, MCS), model group (methionine-choline-deficiency diet, MCD) and intervention group [MCD+1.25(OH) (2)D(3)]. The intervention, control, and model group was given 3 ng/100 g 1.25(OH) (2)D(3) peanut oil solution per day by gavage according to body mass. After 4 weeks the experiment was ended up, and the blood was collected from the inferior vena cava to detect alanine aminotransferase (ALT) and aspartate aminotransferase (AST). The liver tissue was collected to observe the liver morphological and pathological changes (oil red O and HE staining). The changes in the level of liver total triglyceride (TG) content and liver lipid metabolism-related genes [fatty acid transfer protein (FAT/CD36), acetyl-coenzyme A carboxylase (ACC1)] mRNA and protein were detected. One-way analysis of variance was used to compare the means between groups. Results: Oil red O staining and HE staining showed that lipid droplet-vacuoles were significantly increased in the liver tissue of the model group than that of the intervention group. The liver TG content (2.23 ± 0.98) µmol/g of the intervention group was significantly lower than that of the model group (3.53 ± 1.06) µmol/g (F = 5.930, P = 0.035). The ALT content of the intervention group (35.99±9.54) U/L was significantly lower than that of the model group (57.65 ± 19.42) U/L (F = 13.790, P = 0.034). The AST content of the intervention group (16.9 ± 3.73) U/L was significantly lower than that of the model group (27.81 ± 13.31) U/L (F = 3.084, P = 0.046). The relative expression levels of mRNA and protein (mRNA: 1.21 ± 0.61, protein: 1.54 ± 0.75) of FAT/CD36 in the intervention group were significantly lower than those of the model group (mRNA: 2.31 ± 0.81, protein: 2.83 ± 1.42) (mRNA: F = 8.370, P = 0.001, protein: F = 7.212, P = 0.043). The relative expression level of mRNA and protein of ACC1 (mRNA: 0.89 ± 0.54, protein: 0.28 ± 0.11) were also significantly lower than those in model group (mRNA: 1.39 ± 0.19, protein: 0.47 ± 0.24) (mRNA: F = 3.948, P = 0.036, protein: F = 10.933, P = 0.048). Conclusion: 1.25(OH) (2)D(3) can reduce liver fat deposition in rats fed with MCD by inhibiting the expression of fat / CD36 and ACC1.

[Progress of researches on the interaction between parasitic infections and host cell autophagy].

Autophagy, a conserved intracellular degradation system, is a specific life phenomenon in eukaryocytes. Autophagy is widely accepted as a pathway that double-membrane autophagosomes envelop and sequester intracellular cytoplasmic components and then fuse with lysosomes to form autolysosomes, which degrade their contents to regenerate nutrients. Autophagy may be triggered by starvation and a diverse range of pathogens, including parasites. Following infection with intracellular parasites, host cells may eliminate parasites by autophagy. However, parasites may develop self-defense mechanisms, and promote the self-growth and -development by host cell autophagy. This review describes the advances in the interplay between parasitic infections and host cell autophagy. Understanding autophagy is of great significance for the management of parasitic infections and the development of antiparasitic drugs.

MicroRNA-34a suppresses neuronal apoptosis and alleviates microglia inflammation by negatively targeting the Notch pathway in spinal cord injury.

OBJECTIVE: The purpose of this study was to investigate role of inhibition of microRNA-34a (miR-34a) in neural damage and repair after spinal cord injury, and to explore the underlying mechanism. MATERIALS AND METHODS: In BV2 microglia, we conducted classical activation using lipopolysaccharide (LPS) and pre-treatment using miR-34a mimics. The expressions of miR-34a, Notch 1, and Jagged 1 were detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). Moreover, the protein expressions of inflammatory microglia markers were evaluated by Western blotting. In vivo, SCI model was successfully established in rats. Subsequently, the expression levels of miR-34a, Notch 1, and Jagged 1 levels within 1 week were measured by qRT-PCR. Meanwhile, protein expressions of inflammatory mediators were determined by enzyme-linked immunosorbent assay (ELISA) assay. Immunofluorescence was conducted to display the activation degree of microglia and residual neural structure. Furthermore, locomotor function recovery was estimated using BBB rating scale. RESULTS: Compared with the only LPS-activated group, pre-treatment of miR-34a mimics significantly decreased the expressions of Notch 1 and Jagged 1. Similarly, the protein expressions of CD11b and iNOS were significantly down-regulated. In vivo, the levels of Notch 1 and Jagged 1 within 1 week increased significantly, while miR-34a was negatively regulated following spinal cord injury (SCI). Furthermore, the contents of interleukin-1 beta (IL-1ß) and IL-6 were reduced with the treatment of miR-34a mimics when compared with SCI group. With the treatment of miR-34a, the number of inflammatory microglia decreased significantly, and the remaining neural structure was similarly improved. In addition, locomotor function recovery of hindlimbs in rats was significantly ameliorated after the administration of miR-34a mimics. CONCLUSIONS: Increase of miR-34a suppresses neuronal apoptosis and alleviates microglia inflammation by negatively targeting the Notch pathway, thereby improving neural recovery and locomotor function.

Reduction in inappropriate MRI knee studies after implementation of an appropriateness checklist: Experience at a tertiary care centre.

PURPOSE: To evaluate reduction in inappropriate knee MRI requests following implementation of a mandatory knee MRI appropriateness checklist. METHODS: A retrospective review was performed at a single tertiary care centre. A knee MRI appropriateness checklist was developed based on the ACR Appropriateness Criteria and adherence from referring physicians was mandatory. Reports from 200 consecutive knee MRI studies one year prior to implementation were compared to 200 consecutive knee MRI studies following implementation. The presence of moderate or greater osteoarthritis on MRI reports was used as a marker for inappropriate knee MRIs. Patient demographics, wait times, number of knee MRIs, and number of all MRIs at our centre over a six month period post-intervention and pre-intervention were recorded. Differences between pre-intervention and post-intervention presence of moderate or greater osteoarthritis, patient demographics, wait times, and number of MRIs analyzed. RESULTS: A significant decrease was found in moderate or greater grade osteoarthritis following intervention, decreasing from 36.5% to 20.5% (73 studies versus 41 studies, p = 0.023). Of these, the most profound decrease occurred in studies with severe osteoarthritis, with an 80 % decrease (35 studies versus 7 studies, p < 0.001). Post intervention, 48 % fewer knee MRIs were performed in the same time interval (652 studies pre-intervention versus 336 studies post intervention, p < 0.001). No significant differences were found in the patient demographics. CONCLUSION: Mandatory knee MRI appropriateness checklists are associated with a significant reduction in the number of inappropriate studies performed. Follow up studies will be required to assess long-term impact in a larger population.

[Implication of BRAF V600E and CTNNB1 gene mutations in the pathological classification of craniopharyngioma].

Objective: To investigate the clinicopathological significance of BRAF V600E and CTNNB1 gene mutations in adamantinomatous craniopharyngiomas (ACP) and papillary craniopharyngiomas (PCP). Methods: The retrospective study included a total of 67 craniopharyngiomas diagnosed from October 2009 to August 2018 at Xuanwu Hospital, Capital Medical University. The immunohistochemical staining for ß-catenin and BRAF V600E expression, Sanger sequencing of exon 3 of CTNNB1, BRAF mutation analysis by scorpions amplification refractory mutation system (ARMS) fluorescence quantitative PCR were performed. Univariate survival analysis was used to correlate with tumor recurrence. Results: Of the 67 patients, 53 were ACPs and 14 were PCPs. Four patients underwent multiple operations and one of them presented with malignant transformation into squamous cell carcinoma. Histologically, ACPs were characterized by whorl-like cell clusters, peripheral palisaded layer, stellate reticulum, finger-shaped protrusions, ghost cells and wet keratinous substances. While PCPs usually consisted of mature squamous epithelium associated with fibrovascular stroma resulting in papillary appearance. The nuclear immunopositivity for ß-catenin was observed in 73.6% (39/53) of ACPs, and it was absent in PCPs (0/14). The nuclear translocation of ß-catenin usually presented at whorl-like structures or around ghost cells. Of all the cases, mutations analysis in exon 3 of ß-catenin gene CTNNB1 were successful in 46 cases and 42.1% (16/38) of ACP showed CTNNB1 gene mutation, while none of the PCPs harbored CTNNB1 gene mutation (0/8). The cytoplasmic immunopositivity for BRAF V600E mutant protein was found in all PCPs (14/14) and negative in all ACPs (0/53). ARMS-PCR results showed that BRAF V600E mutations were observed in 13/14 of PCPs but not seen in ACPs (0/53). Follow-up data were available in 35 patients with duration of 2 to 120 months. Ten patients experienced recurrences after the first surgery. Upon univariate survival analysis, only subtotal excision was found to be associated with increased recurrence (P=0.032), while pathological type, postoperative radiotherapy and CTNNB1 gene mutation were not (P>0.05). Conclusions: There is significant difference in the expression of BRAF V600E and CTNNB1 genes between ACP and PCP, and their immunohistochemical and molecular detection therefore can be used in the diagnosis and differential diagnoses of craniopharyngiomas.

[Short-term effects of air pollution on lung function of school-age children in Hangzhou].

A total of 1 685 school-age children selected from Hangzhou received lung function testing to evaluate the short-term effects of air pollution on their lung function. The results showed that in every 10 µg/m(3) increase of average concentration of PM(2.5) and PM(10) on the day of the test and the day before the test,peak expiratory flow (PEF) decreased 0.039 (95%CI: 0.012-0.067) L/s and 0.031 (95% CI:0.011-0.051) L/s,respectively. When the average concentration of SO(2) increased 10 µg/m(3) on the day of test and the day prior to the test, PEF and 75% of the forced vital capacity that has not been exhaled (MEF(75)) decreased 0.437 (95%CI: 0.217-0.658) L/s and 0.396 (95%CI: 0.180-0.613) L/s. After being adjusted for NO(2),with every 10 µg/m(3) increase of average concentration of PM(2.5) and PM(10) on the day of the test and the day before the test,PEF and MEF(75) decreased 0.056 (95%CI: 0.028-0.085), 0.053(95%CI: 0.027-0.081) and 0.047 (95%CI: 0.026-0.068) L/s,0.044 (95%CI: 0.023-0.065) L/s on the day before the test, respectively. The results indicate that air pollution have short-term and lag effects on lung function of school-age children in Hangzhou.

[Correlation between prognosis and laboratory parameters of complete deafness in patients with sudden deafness].

Objective: To study the correlation between the prognosis of sudden deafness and laboratory indicators, and to provide a theoretical basis for clinical evaluation of prognosis and selection of appropriate interventions.Method: A retrospective analysis of 123 cases of total deafness was conducted(all frequencies decreased with 250-8 000 Hz average hearing threshold ≥ 81 dB HL). All patients admitted to the hospital received laboratory test within 24 hours and underwent a unified treatment: alprostadil + mecobalamin + citicoline sodium phosphate + Ginkgo biloba extract + batroxobin + methylprednisolone (patients with hypertension or diabetes were given retro-auricular injection). After 2 weeks of comprehensive treatment, the pure tone audiometry was reviewed and the clinical efficacy was evaluated. According to the prognosis, the patients were divided into the healing group, the marked effective group, the effective group and the ineffective group. The laboratory indicators included: white blood cell count, neutrophil percentage, neutrophil lymphocyte ratio(NHR), erythrocyte sedimentation rate, blood lipid, red blood cell count, hemoglobin(Hb), plasma fibrinogen content(FIB). Result: There were no significant differences in white blood cell count, neutrophil percentage, NHR, erythrocyte sedimentation rate, blood lipid index and red blood cell count between different prognosis groups(all P>0.05). The differences of FIB and Hb were statistically significant(P<0.05). The overall prognosis was negatively correlated with FIB and positively correlated with Hb. Patients with vertigo had a worse prognosis and a higher plasma FIB level than those without vertigo(P<0.05), but there was no significant difference of Hb between patients with and without vertigo(P>0.05). Conclusion: FIB and Hb can be used as correlative indexes to evaluate the prognosis of patients with sudden deafness, and the effects of vertigo and other associated symptoms on the prognosis should be comprehensively analyzed. .

Drug-resistance reversal in colorectal cancer cells by destruction of flotillins, the key lipid rafts proteins.

Multi-drug resistance (MDR) of tumor cells attenuates the efficacy of anticancer drugs and has become the main reason for chemotherapy failure. It is indispensable to establish an effective way to reverse multi-drug resistance. Our previous work has shown that down-regulation of the ERK/MAPK signaling pathway activity can reverse the drug-resistance of resistant cells. Further-more, the effect of signal transduction is strongly associated with lipid rafts. The drug-resistance is reversed successfully after lipid rafts are destroyed by heptakis(2, 6-di-O-methyl)-ß-cyclodextrin (MßCD). However, the reversal of the drug-resistance is not associated with down-regulation of the expression of ERK1/2. Cell membrane permeability may increase when lipid rafts are destroyed by MßCD, causing the reversal of drug-resistance due to an increase in accumulation of the drugs in the cytoplasm. To minimize the influence of MßCD on the cell membrane structure, we selected flotillin, a marker protein of lipid rafts, as the target molecule, to further investigate the mechanism of changes in drug resistance after destruction of the lipid rafts. The effect of flotillin on the reversal of the drug resistance was examined using an RNA interference (RNAi) in a retrovirus system in human drug-resistant strains of colorectal cancer cell line HCT-15. The results demonstrate that flotillin-1 downregulation by RNAi (Flot1-RNAi) reduced the drug resistance, caused cell cycle arrest and decreased the expression of ERK1/2; however, apoptosis was not significantly affected. Knockdown of flotillin-2 by RNAi (Flot2-RNAi) had effects similar to those of Flot1-RNAi except that the effects on expression of ERK1/2 and apoptosis were different. Screening of multiple pathways indicated that the PI3K/Akt signaling pathway was closely related. This experiment demonstrates the association between PI3K and drug resistance through the activation of PI3K and suggests that PI3K may play a key role during the development of resistance in CRC. The results reveal that the levels of IRS-1 and PI3K proteins in the Flot1-RNAi and Flot2-RNAi groups were significantly down-regulated. Knockdown of flotillins by RNAi reduced the resistance of HCT-15/ADM cells; the results investigations of the Akt pathway indicate a decrease in resistance after lipid raft destruction. These data confirm that knockdown of flotillin reduces the resistance of HCT-15/ADM cells, and the mechanism may be relevant to the PI3K/Akt pathway. Additionally, flotillin may be used as a potential target for chemotherapy in the treatment of colorectal cancer.

[Three-dimensional finite element analysis of four-implants supported mandibular overdentures using two different attachments].

Objective: To compare the biomechanical characteristics of four-implants mandibular overdentures supported by Locator attachment or bar-clip attachment under different mechanical loads using three-dimensional finite element analysis method. Methods: Two different models of four-implants supported mandibular overdentures using Locator attachment and bar-clip attachment (hereinafter called Locator model and bar-clip model) were established. Each model was subjected to five different mechanical loading conditions: 100 N vertical loading in central incisor (vertical load of incisor), 100 N vertical loading or oblique loading in canine (vertical or oblique loads of canines), 100 N vertical or oblique loading in mandibular first molar (vertical or oblique loads of mandibular first molar). The stress distributions in implants, peri-implant bone and mucosa were recorded under the above five conditions to evaluate the effects of different attachments on the biomechanical properties of implant-supported mandibular overdentures. Results: Regardless of loading conditions and types of attachments, the stress concentration in implants were located at the neck of implants, and the stress concentration in peri-implant bone was located in the cortical bone. The stress values in mucosa were always much smaller than those in implants and cortical bone. Regardless of loading positions (on canine or on mandibular first molar), the maximum stress at the bone interface around the implant under lateral loading was much higher than that under vertical loading. Under various loading conditions, the stress in implants and cortical bone of the Locator model (the highest von Mise stress value was respectively 79.5 and 22.3 MPa) were lower than that of bar-clip model (the highest von Mise stress value was 110.3 and 28.7 MPa respectively) while the maximum compressive stress in mucosa (0.198 MPa) in Locator model was slightly higher than that in the bar-clip model (0.137 MPa). Conclusions: In clinical practice, the lateral force applied to the implant-retained overdenture should be minimized to avoid complications caused by pathological loads. Under the same loading condition, the stress distributions in overdenture using Locator attachment are more dispersed, which is more conducive to long-term stability of implants.

Arginine bioavailability and endothelin-1 system in the regulation of vascular function of umbilical vein endothelial cells from intrauterine growth restricted newborns.

BACKGROUND AND AIMS: Intrauterine growth restriction (IUGR) is a major risk factor for perinatal morbidity and mortality, leading to long-term adverse cardiovascular outcomes. The present study aimed to investigate the potential mechanisms in IUGR-associated vascular endothelial dysfunction. METHODS AND RESULTS: Human umbilical vein endothelial cells (HUVECs) were derived from IUGR or normal newborns. We found that the proliferation of IUGR-derived HUVECs was accelerated compared to those from normal subjects. Gene profiles related to vascular function including vasomotion, oxidative stress, and angiogenesis were dysregulated in IUGR-HUVECs. Compared with HUVECs from normal newborns, nitric oxide (NO) production was reduced, with imbalance between endothelial nitric oxide synthase (eNOS) and arginase-2 (Arg-2) in IUGR. Meanwhile, intracellular asymmetric dimethylarginine (ADMA) level was elevated with diminished dimethylarginine dimethylaminohydrolase 1 (DDAH1) expression in IUGR-HUVECs. Furthermore, endothelin-1 (ET-1) and hypoxia-inducible factor 1α (HIF-1α) expression were increased, and endothelin receptor type-B (ETBR) was reduced in the IUGR group. IUGR-HUVECs exposed to hypoxia increased the ratio of ADMA to l-arginine, HIF-1α and protein arginine methyltransferase 1 (PRMT1) expression compared to controls. CONCLUSIONS: The present study demonstrated that the reduction of NO bioavailability and release results from elevated Arg-2, accumulation of intracellular ADMA, and imbalance of ET-1 and ETBR, further leading to IUGR-associated vascular endothelial dysfunction. Our study provides novel evidence on the mechanism underlying fetal programming associated with IUGR, which will serve as potential therapeutic targets in the prevention of adverse cardiovascular consequences in adulthood.

Genome-wide association study reveals novel genes for the ear size in sheep (Ovis aries).

Variations in ear size can be observed in livestock such as sheep; however, the genetic basis of variable ear size in sheep is still poorly understood. To investigate causative genes associated with ear size in sheep, a genome-wide association study was performed in 115 adult Duolang sheep with different-sized floppy ears using the Ovine Infinium HD BeadChip. We found 38 significant SNPs at the genome-wide or chromosome-wise 5% significance level after Bonferroni correction. The most significant association (P = 1.61 × 10-6 ) was found at SNP rs402740419, located in the DCC gene, which plays a critical role in ear development. Also, we observed two additional significant SNPs, rs407891215 in PTPRD and rs407769095 in SOX5, both of which are functionally associated with ear developmental processes. Our results are useful for future sheep breeding and provide insights into the genetic basis of ear size development in sheep and other livestock.

A genome-wide association study reveals candidate genes for the supernumerary nipple phenotype in sheep (Ovis aries).

Genome-wide association studies (GWASs) have been widely applied in livestock to identify genes associated with traits of economic interest. Here, we conducted the first GWAS of the supernumerary nipple phenotype in Wadi sheep, a native Chinese sheep breed, based on Ovine Infinium HD SNP BeadChip genotypes in a total of 144 ewes (75 cases with four teats, including two normal and two supernumerary teats, and 69 control cases with two teats). We detected 63 significant SNPs at the chromosome-wise threshold. Additionally, one candidate region (chr1: 170.723-170.734 Mb) was identified by haplotype-based association tests, with one SNP (rs413490006) surrounding functional genes BBX and CD47 on chromosome 1 being commonly identified as significant by the two mentioned analyses. Moreover, Gene Ontology enrichment for the significant SNPs identified by the GWAS analysis was functionally clustered into the categories of receptor activity and synaptic membrane. In addition, pathway mapping revealed four promising pathways (Wnt, oxytocin, MAPK and axon guidance) involved in the development of the supernumerary nipple phenotype. Our results provide novel and important insights into the genetic mechanisms underlying the phenotype of supernumerary nipples in mammals, including humans. These findings may be useful for future breeding and genetics in sheep and other livestock.

Genome-wide association analysis identifies the genetic basis of fat deposition in the tails of sheep (Ovis aries).

Fat-tailed sheep (Ovis aries) can survive in harsh environments and satisfy human's intake of dietary fat. However, the animals require more feed, which increases the cost of farming. Thus, most farmers currently prefer thin-tailed, short-tailed or docked sheep. To date, the molecular mechanism of the formation of fat tails in sheep has not been completely elucidated. Here, we conducted a genome-wide association study using phenotypes and genotypes (the Ovine Infinium HD SNP BeadChip genotype data) of two breeds of contrasting tail types (78 Small-tailed and 78 Large-tailed Han sheep breeds) to identify functional genes and variants associated with fat deposition. We identified four significantly (rs416433540, rs409848439, rs408118325 and rs402128848) and three approximately associated autosomal SNPs (rs401248376, rs402445895 and rs416201901). Gene annotation indicated that the surrounding genes (CREB1, STEAP4, CTBP1 and RIP140, also known as NRIP1) function in lipid storage or fat cell regulation. Furthermore, through an X-chromosome-wide association analysis, we detected significantly associated SNPs in the OARX: 88-89 Mb region, which could be a strong candidate genomic region for fat deposition in tails of sheep. Our results represent a new genomic resource for sheep genetics and breeding. In addition, the findings provide novel insights into genetic mechanisms of fat deposition in the tail of sheep and other mammals.

[Progress of sino-nasal IgG4-related disease].

IgG4-related disease is a newly recognized systemic fibro inflammatory disorder that affects the sino-nasal region. It is a rare and emerging entity that can present with bony and soft-tissue invasion,the final diagnosis of this disease mainly depends on pathological examination and majority of patients receiving corticosteroids responded very well to treatment. Thus,Our goal was to highlight the sino-nasal presentation of this unique disease and to review previously reported cases from 2010 to 2016.We hope that clinical physicians to enhance understanding of the disease in order to ensure early diagnosis and early intervention to prevent serious injury and fibrosis of organs.

An innovative SNP genotyping method adapting to multiple platforms and throughputs.

KEY MESSAGE: An innovative genotyping method designated as semi-thermal asymmetric reverse PCR (STARP) was developed for genotyping individual SNPs with improved accuracy, flexible throughputs, low operational costs, and high platform compatibility. Multiplex chip-based technology for genome-scale genotyping of single nucleotide polymorphisms (SNPs) has made great progress in the past two decades. However, PCR-based genotyping of individual SNPs still remains problematic in accuracy, throughput, simplicity, and/or operational costs as well as the compatibility with multiple platforms. Here, we report a novel SNP genotyping method designated semi-thermal asymmetric reverse PCR (STARP). In this method, genotyping assay was performed under unique PCR conditions using two universal priming element-adjustable primers (PEA-primers) and one group of three locus-specific primers: two asymmetrically modified allele-specific primers (AMAS-primers) and their common reverse primer. The two AMAS-primers each were substituted one base in different positions at their 3' regions to significantly increase the amplification specificity of the two alleles and tailed at 5' ends to provide priming sites for PEA-primers. The two PEA-primers were developed for common use in all genotyping assays to stringently target the PCR fragments generated by the two AMAS-primers with similar PCR efficiencies and for flexible detection using either gel-free fluorescence signals or gel-based size separation. The state-of-the-art primer design and unique PCR conditions endowed STARP with all the major advantages of high accuracy, flexible throughputs, simple assay design, low operational costs, and platform compatibility. In addition to SNPs, STARP can also be employed in genotyping of indels (insertion-deletion polymorphisms). As vast variations in DNA sequences are being unearthed by many genome sequencing projects and genotyping by sequencing, STARP will have wide applications across all biological organisms in agriculture, medicine, and forensics.

Identification of a potential functional single nucleotide polymorphism for fatness and growth traits in the 3'-untranslated region of the PCSK1 gene in chickens.

Prohormone convertase 1/3 is a serine endoprotease belonging to the subtilisin-like proprotein convertase family that is encoded by the () gene, and its major function is the processing and bioactivation of the proproteins of many kinds of neuroendocrine hormones, including insulin, cholecystokinin, and adrenocorticotropic hormone. The results of our previous genomewide association study indicated that the gene might be an important candidate gene for fatness traits in chickens. The objectives of this study were to investigate the tissue expression profiles of gene and to identify functional variants associated with fatness and growth traits in the chicken. The results indicated that mRNA was widely expressed in various tissues, especially neuroendocrine and intestinal tissues. Of these 2 tissue types, mRNA expression in lean males was significantly higher than in fat males. A SNP in the 3' untranslated region of (c.*900G > A) was identified. Association analysis in the Arbor Acres commercial broiler population and Northeast Agricultural University broiler lines divergently selected for abdominal fat content (NEAUHLF) population showed that the SNP c.*900G > A was associated with abdominal fat weight, abdominal fat percentage, BW, metatarsus length, and metatarsal circumference. In the 5th to 19th generation (G to G) of NEAUHLF, the allele frequency of c.*900G > A changed along with selection for abdominal fat content. At G, allele G of c.*900G > A was predominate in the lean line, whereas allele A was predominate in the fat line. Functional analysis demonstrated that allele A of c.*900G > A reduced mRNA stability and consequently downregulated gene expression. These results suggested that c.*900G > A was a functional SNP for fatness and growth traits in the chicken. The results of this study provide basic molecular information for the role of gene in avian growth and development, especially obesity.

Expression analysis of bone morphogenetic protein 4 between fat and lean birds in adipose tissue and serum.

The objectives of the present study were to characterize the tissue expression of chicken (Gallus gallus) bone morphogenetic protein 4 (BMP4) and compare differences in its expression in abdominal fat tissue and serum between fat and lean birds and to determine a potential relationship between the expression of BMP4 and abdominal fat tissue growth and development. The results showed that chicken BMP4 messenger RNA (mRNA) and protein were expressed in various tissues, and the expression levels of BMP4 transcript and protein were relatively higher in adipose tissues. In addition, the mRNA and protein expression levels of BMP4 in abdominal fat tissue of fat males were lower than those of lean males at 1, 2, 5, and 7 wk of age (P < 0.05). Furthermore, the serum BMP4 content of fat males was lower than that of lean males at 7 wk of age (P < 0.05). BMP4 mRNA expression levels were significantly higher in preadipocytes than those in mature adipocytes (P < 0.05), and the expression level decreased during differentiation in vitro (P < 0.05). These results suggested that chicken BMP4 might affect abdominal fat deposition through differences in its expression level. The results of this study will provide basic molecular information for studying the role of BMP4 in the regulation of adipogenesis in avian species.

Serum microRNA-210 as a potential noninvasive biomarker for the diagnosis and prognosis of glioma.

BACKGROUND: MicroRNA-210 (miR-210) is an oncogenic miRNA previously associated with prognosis in human gliomas, an incurable tumour type of the central nervous system. Here miR-210 was investigated as a potential serum biomarker in the diagnosis and prognosis of glioma. METHODS: Serum was immediately prepared from blood samples collected from patients with glioma grades I-IV at primary diagnosis (n=136) and healthy controls (n=50) from February 2007 to March 2014 in the Department of Neurosurgery of the First Affiliated Hospital of Wannan Medical College (Wuhu, China). Total RNA was isolated from serum. cDNA was synthesised with primers specific for miR-210 and miR-16-1 (internal control), and quantitative real-time RT-PCR was performed. Results were statistically analysed to determine the role of miR-210 in the diagnosis and prognosis of human glioma patients. RESULTS: An approximately seven-fold increase in miR-210 expression was detected in serum samples from glioblastoma patients relative to healthy controls. A threshold expression value (2.259) was chosen from receiver operator characteristic curves (ROC), and the low and high miR-210 expression groups were analysed by multivariate Cox proportional hazard regression and Kaplan-Meier analyses. Results revealed an association of high serum miR-210 expression with tumour grade and poor patient outcome (P-values <0.001). CONCLUSIONS: Serum miR-210 is a promising diagnostic and prognostic biomarker that can be detected in the peripheral blood of patients with glioma.