RBM46 is a germ cell-specific RNA-binding protein required for gametogenesis, but the targets and molecular functions of RBM46 remain unknown. Here, we demonstrate that RBM46 binds at specific motifs in the 3'UTRs of mRNAs encoding multiple meiotic cohesin subunits and show that RBM46 is required for normal synaptonemal complex formation during meiosis initiation. Using a recently reported, high-resolution technique known as LACE-seq and working with low-input cells, we profiled the targets of RBM46 at single-nucleotide resolution in leptotene and zygotene stage gametes. We found that RBM46 preferentially binds target mRNAs containing GCCUAU/GUUCGA motifs in their 3'UTRs regions. In Rbm46 knockout mice, the RBM46-target cohesin subunits displayed unaltered mRNA levels but had reduced translation, resulting in the failed assembly of axial elements, synapsis disruption, and meiotic arrest. Our study thus provides mechanistic insights into the molecular functions of RBM46 in gametogenesis and illustrates the power of LACE-seq for investigations of RNA-binding protein functions when working with low-abundance input materials.
Animals ; Mice ; 3' Untranslated Regions/genetics* ; Cell Cycle Proteins/metabolism* ; Gametogenesis/genetics* ; Meiosis/genetics* ; Nuclear Proteins/genetics* ; RNA-Binding Proteins/genetics*

Objective To establish a reliable pretreatment method for the detection of mequindox and its metabolite in pork luncheon meat by ultra-performance liquid chromatography/triple qudrupole tandem mass spectrometry. Methods Samples were extracted with ethyl acetate, and the results of purification and enrichment by PAX and PEP solid-phase extraction columns were analyzed. Acetonitrile/methanol (3:11) - 0.1% formic acid water was used as the mobile phase, and Shimadzu Inertsil ODS-3-column (3µm, 2.1 × 100mm) chromatographic columns were used for qualitative and quantitative analysis using the multi-reaction detection positive ion mode. Results The results showed that PEP cartridge had good recovery rate. The detection limit of mequindox was 0.10µg/kg, and limit of quantitation was 0.30µg/kg. The average recoveries for spiked levels of 0.33, 0.83, and 1.65µg/kg were 127%, 72.0%, and 60.1%, respectively. The detection limit of 2-quinoxalinecarboxylic acid was 0.10µg/kg, and limit of quantitation was 0.40µg/kg. The average recoveries for spiked levels of 0.42, 1.05, and 2.1µg/kg were 125%, 99.0%, and 60.9%, respectively. Conclusion This method is suitable for the determination of mequindox and its metabolite 2-quinoxalinecarboxylic acid in luncheon meat.

Objective:To optimize the extraction process of Shangke Zhitong oil. Methods:Taking the comprehensive score of extracting amounts of acteoside and garlic acid as the index, L9(34)orthogonal test was used to observe the effects of alcohol concentration,soaking time, extraction times and extraction duration on the extraction results. Results:The best extraction process was as follows:the herbs were soaked for 1 h in 65% alcohol,and then refluxed and extracted for three times with 1.5 h for the first time,1 hour for the second time and the third time. The extraction capacity of acteoside and garlic acid was 0.870 8 and 0.717 8 mg·ml-1,respectively. Conclusion:The extraction process is simple,feasible,scientific and reasonable.

OBJECTIVETo analyze two fetuses with multiple malformations revealed by ultrasonography using single nucleotide polymorphism array (SNP array), and to explore the strategy for the prenatal diagnosis of 1p36 deletion syndrome.

METHODSAmniocentesis was performed on the two pregnant women. Amnion fluid cells were cultured, and karyotypes of the fetuses were determined through G-banding analysis. Whole genome SNP array was used to detect genomic anomalies of the two fetuses. The karyotypes of their parents were determined through G-banding analysis of peripheral venous blood samples.

RESULTSG-banding analysis showed a 46,XY,add(1p36)? and a 46,XX,add(1p36)? karyotype for fetuses 1 and 2, respectively. SNP array analysis showed that the fetus 1 had arr[19]1p36.33p36.32 (752 566 - 3 393 462)×1 and 7q35q36.3 (144 480 549 - 159 119 486)×3, and fetus 2 had arr[19]1p36.33p36.23 (752 566 - 8 362 754)×1, 6p25.3p22.3 (204 909 - 20 182 185)×3. The mother of fetus 1 had a 46,XX,t(1;7)(p36;q35) karyotype, and the mother of fetus 2 had a 46,XX,t(1;6)(p36;p22) karyotype. The karyotypes of both fathers appeared to be normal.

CONCLUSIONSNP array has the advantages such as high sensitivity and high accuracy for prenatal diagnosis, and can provide more detailed information for genetic counseling of 1p36 deletion syndrome.

Adult ; Amniocentesis ; Chromosome Banding ; Chromosome Deletion ; Chromosome Disorders ; diagnosis ; Chromosomes, Human, Pair 1 ; Female ; Humans ; Karyotyping ; Polymorphism, Single Nucleotide ; Pregnancy ; Prenatal Diagnosis

Objective To explore the association of Crohn's disease (CD) with T cell immunoglobulin and mucin domain 3 (Tim-3) gene polymorphisms in patients of Zhejiang Han population in China.Methods A total of 308 CD patients and 573 age-and sex-matched healthy controls were enrolled in our study.Two single nucleotide polymorphisms (SNPs) of Tim-3 (rs1036199 and rs10515746) were examined by the improved multiple ligase detection reaction technique (iMLDR).Analyses of linkage disequilibrium and haplotype were also performed by Haploview 4.2 software in all study subjects.Results In general,the allele and genotype frequencies of Tim-3 (rs1036199 and rs10515746) were not statistically different between CD patients and the controls (all P ＞0.05).According to the Montreal Classification,CD patients were divided into different subgroups.The variant allele (C) and genotype (AC + CC) of rs1036199 were more frequent in CD patients with penetrating diseases than in the controls (10.4％ vs 1.7％,P =0.002;20.8％ vs 3.5％,P =0.023).Similar conclusions were also drawn for the variant allele (A) and genotype (CA + AA) of rs10515746 in patients with penetrating diseases when compared with the controls (10.4％ vs 2.2％,P =0.000;20.8％ vs 4.2％,P =0.033,respectively).The two SNPs of Tim-3 were in strong linkage disequilibrium (D'=1.0,r2 =0.928).The haplotype (AC) formed by their wild-type alleles (A) and (C) was decreased in patients with penetrating CD compared with the controls (89.6％ vs 98.3％,P =0.000).However,the haplotype (CA) formed by their variant alleles was more frequent in patients with penetrating CD than in the controls (10.4％ vs 1.6％,P =0.000).Conclusions Tim-3 (rs1036199 and rs10515746) variations might be correlated with the enhanced risk of penetrating diseases in CD patients.Furthermore,the haplotype (AC) and (CA) formed by the two SNPs might be a protective and a risky factor for penetrating CD respectively.

Objective To analyze the clinical features,diagnosis and treatment of adult Moyamoya disease in countryside of the southeast of Hubei province.Methods Sixty-eight adult patients with Moyamoya disease,selected in countryside of the southeast of Hubei Province from May 2010 to May 2015,were enrolled.The clinical data,including age,gender,address (surrounding of residence),living habit (special hobby),health of family members,past medical history,symptom,confirmation related factors,treatment methods and prognoses,were retrospectively analyzed.Results These patients had high and low incidences in distribution.The ratio of male to female was 1.09:1.The peak age of onset was 35 to 44 years.There were 42 bleeding patients,16 ischemia patients,and 7 patients with atypical symptom,and 3 patients were asymptomatic.Thirty-eight patients (55.9％) had early diagnosis and 30 (44.1％) had late conformed diagnosis;patients with early diagnosis had significantly higher percentages of first diagnosis in the tertiary hospitals,hemorrhagic apoplexy as first onset,high education level and economic level than patients with late conformed diagnosis (P＜0.05).Ten patients died.Eight patients received vascular reconstruction,accounting for 11.8％ (8/58).Conclusions The distributions of adult moyamoya disease are regional cluster.The main age of onset is at the life prime.The initial symptom is hemorrhagic stroke.The time for diagnosis was short in the tertiary hospitals.The ratio of patients receiving vascular reconstruction is low.

OBJECTIVETo develop and validate a method for mutation screening and prenatal diagnosis of TSC1/TSC2 mutations among patients with tuberous sclerosis complex (TSC) by Ion Torrent semiconductor sequencing.

METHODSPotential mutations of SC1/TSC2 gene was detected in 2 TSC families and 1 sporadic TSC patient using an Ion Torrent PGM sequencer. Candidate variants were validated by Sanger sequencing. The corresponding site of TSC2 in the fetus of family 2 was also detected with Sanger sequencing.

RESULTSIon Torrent semiconductor sequencing has identified a probably pathogenic TSC2 mutation (c.311-312insGCTG) in the patient from family 1, and a probably pathogenic TSC2 mutation (c.1790A>G) in the patient of family 2.

CONCLUSIONTargeted Ion Torrent PGM sequencing is an accurate and efficient method to detect TSC1/TSC2 mutations in TSC.

Adult ; Base Sequence ; DNA Mutational Analysis ; Female ; Humans ; Male ; Molecular Sequence Data ; Mutation ; Pedigree ; Pregnancy ; Prenatal Diagnosis ; Tuberous Sclerosis ; embryology ; genetics ; Tumor Suppressor Proteins ; genetics ; Young Adult

Objective To probe into the clinical nurse′s ethical cognition of Kazakh Medicine Hospital of Altay Region in Xinjiang,so as to provide basis for improving the level of nursing ethical cognition in minority area.Methods The self-designed questionnaire was used to investigate 51 clinical nurses of Kazakh Medicine Hospital of Altay Region in Xinjiang,and the result of investigation was used nonparametric statistics method and was analyzed by software SPSS 16.0.Results The overall level of ethical cognition in 51 clinical nurses maintained low level,with lowest score 30 and highest score 51 and median score 37;there were 21 nurses acquired score range from 30-36,and 30 nurses acquired over 36.The differences were statistically significant in comprehensive ethical cognition score of different departments′nurses (H=16.916,P <0.05).Conclusions The overall level of ethical cognition in 51 clinical nurses was low,so we should pay attention to its education,encourage multi-channel learning,strengthen ethics training,establish ethical norms,narrow the gap among each departments′nurses ethical cognition,improve the overall quality of nurses,and provide quality service for patients.

Objective To investigate associations of UC with the polymorphisms of TRAIL receptors.Methods From January 2008 to December 2012, 380 consecutive UC patients [215 males and 165 females, the average age was (42.63 ±14.61) years] as well as 539 sex-and age-matched healthy individuals [290 males and 249 females, the average age was (41.29 ±15.86) years] were recruited from four large scale comprehensive hospitals in Wenzhou city.Five single nucleotide polymorphisms of DR4 (rs20575, rs13278062), DR5(rs1047266), DcR2(rs1133782) and OPG (rs3102735) were detected by a SNaPshot technique.Distributions of mutant alleles and genotypes for targeted polymorphisms in TRAIL receptors were analyzed by Chi-square test or Fisher′s exact test. By means of unconditional Logistic regression analysis, it evaluated associations between the polymorphisms and the risk of UC attack as well as the clinical features of UC patients.Furthermore, an unconditional Logistic multiple regression analysis was employed to investigate the independent risk factors of UC and their multiplicative interaction effects on UC.Results The frequencies of mutant allele (G) and genotype (CG+GG) of DR4(rs20575) were higher in UC patients than those in the controls (3.55%vs 1.95%,χ2 =4.512, P=0.034;6.58%vs 3.71%,χ2=3.938, P=0.047, respectively).However, the frequeucies of mutant allele ( A) and genotype ( GA+AA) of DcR2(rs1133782) were decreased in UC patients compared to the controls(6.18%vs 9.09%,χ2=5.183, P=0.023; 11.32% vs 17.44%, χ2 =6.589, P=0.010, respectively).The frequencies of mutant allele (T) and homozygote (TT) of OPG(rs3102735) were significantly higher in UC patients than in the controls (86.32% vs 81.54%, χ2 =7.385, P=0.007;75.26% vs 66.98%, χ2 =7.346, P=0.007, respectively) .Furthermore, the genotype (GG) of DcR2 (rs1133782) was found to be the independent risk factor for UC attack (OR=4.937, 95%CI:2.320-10.504, P<0.001).Moreover, the (GG) of DcR2(rs1133782) and (CC) of DR4(rs20575) had an interactive effect on UC (OR=0.322, 95%CI:0.164-0.633, P=0.001).The same conclusion was drawn for the ( GG) of DR4( rs20575) and (TT) of OPG(rs3102735) (OR=1.580, 95%CI:1.165-2.144, P=0.003).Conclusions The genetic polymorphisms of DR4 ( rs20575 ) , DcR2 ( rs1133782 ) and OPG ( rs3102735 ) were associated with UC. The mutation of DcR2(rs1133782) might play a protective role in UC.Moreover, the DcR2(rs1133782) and DR4(rs20575) gene had a collaborative effect on UC.So did the DR4(rs20575) and OPG(rs3102735) genes.

Objective To explore the expression and significance of hsa-miR-181a (miR-181a) in can-ceration progression of endometrial carcinoma. Methods A total of 75 formalin-fixed paraffin-embedded tissue specimens were studied in this study , of which , 13 were normal endometrium , 18 were endometrial hyperplasia , 44 were endometrial carcinoma. After total RNA had been extracted , real-time PCR was applied to detect the ex-pression level of miR-181a in endometrial tissue in each group. Results miR-181a expression in formalin-fixed paraffin-embedded tissue specimens can be detected. Expression of miR-181a in endometrial carcinoma was high-er than that in endometrial hyperplasia , its expression in endometrial hyperplasia was also higher than that in normal endometrium, and the difference was statistically significant (P < 0.05). The expression of miR-181a in endometrial carcinoma was associated with FIGO stages (P < 0.05). Conclusion The up-regulation of miR-181a expression in women with endometrial carcinoma may play the role of oncogenes. Abnormal expression of miR-181a is probably associated with the occurrence and development of endometrial carcinoma.