An ultrasensitive assay format for detecting ULK1 inhibition by monitoring the phosphorylation status of Atg13

A new technology from Quanterix called SiMoA (single molecule array) which employs a fully automated system capable of ultrasensitive sandwich based ELISA detection was explored. Our studies focused upon the inhibition of the autophagy initiating kinase ULK1 by measuring the both total Atg13 and the phosphorylation of Atg13(pSer³¹⁸) from control and following compound treatment in either overexpressing or wild type tissue culture samples. The results show linear protein concentration dependence over two orders of magnitude and provide an assay window of 8- to 100-fold signal to background for inhibition of phosphorylation for both wild type and overexpressed samples, respectively. Moreover, overexpressed samples displayed 17-fold pSer³¹⁸-Atg13 above wild type levels of with no apparent differences in compound potency. Lastly, the inhibition of ULK1 from mouse derived wild type xenografts also demonstrated loss of pSer³¹⁸-Atg13 upon ULK1 inhibitor treatment that compared favorably to Western blot. These results show that the SiMoA technology can detect quantitatively low levels of endogenous biomarkers with the ability to detect the loss of pSer³¹⁸-Atg13 upon ULK1 inhibition.     

Characterization of genes coding for galacturonosyltransferase-like (GATL) proteins in rice

In the present study, seven galacturonosyltransferase-like (GATL) genes (OsGATLs) in rice (Oryza sativa L.) were genome-widely identified and the chromosomal locations and the gene structures of which were characterized. Under normal condition, OsGATL2 and OsGATL3 are highly expressed in root, while OsGATL4 is highly expressed in stem and leaf. Many cis-elements related to stress response and plant hormone were found in the promoter sequence of each OsGATL. The expression patterns of these OsGATL genes under treatment with abscisic acid (ABA), drought and low temperature were assessed by qRT-PCR. The expression levels of most OsGATLs significantly increased following the treatments with drought or low temperature. In addition, physicochemical properties of OsGATLs and phylogenetic analysis with GATL from rice and several other species were performed. 3D structures and protein–protein interaction (PPI) network of OsGATLs were further predicted by Swiss-model and STRING 9.0 database, respectively. The identification and bioinformatic analysis of GATL family in rice could provide reference data for further study on their biological functions, especially in the responsiveness to hormones and stress signaling.     

CpG_MPs: identification of CpG methylation patterns of genomic regions from high-throughput bisulfite sequencing data

High-throughput bisulfite sequencing is widely used to measure cytosine methylation at single-base resolution in eukaryotes. It permits systems-level analysis of genomic methylation patterns associated with gene expression and chromatin structure. However, methods for large-scale identification of methylation patterns from bisulfite sequencing are lacking. We developed a comprehensive tool, CpG_MPs, for identification and analysis of the methylation patterns of genomic regions from bisulfite sequencing data. CpG_MPs first normalizes bisulfite sequencing reads into methylation level of CpGs. Then it identifies unmethylated and methylated regions using the methylation status of neighboring CpGs by hotspot extension algorithm without knowledge of pre-defined regions. Furthermore, the conservatively and differentially methylated regions across paired or multiple samples (cells or tissues) are identified by combining a combinatorial algorithm with Shannon entropy. CpG_MPs identified large amounts of genomic regions with different methylation patterns across five human bisulfite sequencing data during cellular differentiation. Different sequence features and significantly cell-specific methylation patterns were observed. These potentially functional regions form candidate regions for functional analysis of DNA methylation during cellular differentiation. CpG_MPs is the first user-friendly tool for identifying methylation patterns of genomic regions from bisulfite sequencing data, permitting further investigation of the biological functions of genome-scale methylation patterns.     

肺结核合并糖尿病54例临床分析

目的:探讨肺结核合并糖尿病的临床表现及其糖尿病控制对结核病治疗的影响.方法:回顾性分析我院收治的54例肺结核合并糖尿病患者的临床资料.结果:肺结核合并糖尿病以2型糖尿病为主,肺结核X线表现以大片渗出、干酪病变为主,痰菌阳性率高,合并症多为临床特点,血糖大多数控制在7.8mmol/L以下.结论:对肺结核合并糖尿病患者应进行早期正规抗结核治疗,并适当延长疗程,同时有效控制血糖是治疗结核合并糖尿病的关键.     

ICAM-1与脑梗死关系的研究进展

炎症在脑梗死的发病机制中扮演着非常重要的角色,动脉粥样硬化是脑梗死的病理基础,动脉粥样硬化被认为是一种慢性炎症过程,这种炎症过程与黏附分子的表达有关,如细胞间黏附分子-1(ICAM-1),它能黏附循环中的白细胞,促进内皮细胞表面粥样斑块的形成,许多研究表明,ICAM-1与脑梗死密切相关,本文就二者的关系做一综述.     

Field evaluation of juvenile in vitro embryo transfer (JIVET) in sheep

The practicality of using juvenile in vitro embryo transfer (JIVET) on a field scale in China was evaluated in each of three seasons (summer, autumn and winter) from 2006 to 2007. A total of 102 donor Merino lambs (18 summer, 69 autumn and 15 winter) aged 4-8 weeks were stimulated with 4 x 40 mg FSH administered at 12h intervals plus 400 IU PMSG given at the time of the first FSH treatment. Overall, 89.2% (91/102) of the lambs exhibited follicle development and 79.1+/-65.5 (mean+/-S.D.) cumulus-oocyte complexes were recovered per donor lamb. Compared with the groups of summer (84.9+/-55.3) and autumn (83.6+/-70.8) lambs, the number of recovered cumulus-oocyte complexes was significantly decreased in winter (51.4+/-43.7; p<0.05). After recovery, the cumulus-oocyte complexes were matured and fertilized in vitro using frozen-thawed semen and culture in synthetic oviduct fluid medium to the 2-4-c stage of development, when they were transferred surgically in groups of 3-8 (5.33+/-1.47) to the ipsilateral uterine horn of a total of 603 synchronized recipients. The overall mean proportion of cumulus-oocyte complexes developing to 2-c embryos was 61.4% (4308/7013) and differed significantly between seasons (summer 38.5%, autumn 66.1%, winter 74.6%; p<0.01). Pregnancy rate assessed by ultrasound examination approximately 60 days after embryo transfer was 54.4% (328/603) overall, and 36.7% (221/603) of the recipients maintained their pregnancy to full-term, producing an average 1.49 (330/221) offspring, of which 1.21 (267/221) were viable and healthy lambs, per pregnant recipient. Pregnancy rate at day 60 was affected by season (summer 40.5%, autumn 56.7%, winter 55.7%; p<0.05), but did not differ significantly between seasons at full-term (summer 34.2%, autumn 38.9%, winter 30.4%; p>0.05). Based on the number of donors stimulated, the total number of offspring and viable progeny produced per donor lamb in autumn (5.81 and 4.87) was significantly (p<0.01) higher than that of summer (2.79 and 1.94) and winter (4.24 and 3.31). This study showed that each donor lamb after stimulation produced an average of 48.6 transferable embryos that resulted in 4.04 viable and healthy progeny. These results indicate that JIVET is a cost-effective method of multiplying desirable sheep genotypes in China.     

Electrochromatographic characterization of methacrylate ester-based monolith and capillary electrochromatography separation of flavonoids

A porous polymethacrylate ester-based monolithic column for capillary electrochromatography (CEC) was designed by mean of in situ co-polymerizing lauryl methacrylate (LMA), ethylene dimethacrylate (EDMA) and 2-acrylamido-2-methyl-1-propanesulfonic acid (AMPS) in a ternary porogenic solvent including cyclohexanol, 1,4-butanediol and water. After investigating the influence factors of the CEC monolithic columns, four flavonoids (i.e., Rutin, Quercetin, Kaempferol, and Quercitrin) were separated and assayed to evaluate this monolithic column with CEC method. Under optimum conditions, the CEC method exhibited high separation efficiency, with rapid separation time of 3–4 min, for the four flavonoid samples using 10 mM phosphate buffer containing 70% acetonitrile (pH 9.0). Importantly, the proposed method could provide a promising approach for rapid separation and detection in biomedicine.     

银杏雌雄配子体发育及胚胎形成的研究进展

银杏（Ginkgo biloba）是现存最古老的裸子植物之一, 其生殖过程表现出许多原始特征和独特性状, 长期以来备受国内外专家的关注。经过近100年的研究取得了显著成果： （1） 银杏雄配子体发育周期长, 经历了从平周分裂到斜背式分裂,并最终垂周分裂形成带有鞭毛的游动精子; （2） 银杏雌配子体发育经历较长的游离核期和细胞化期, 分化形成颈卵器母细胞并经平周分裂、垂周分裂和斜向分裂形成成熟的颈卵器（包括有4个颈细胞和1个卵细胞）; （3） 推测其精细胞中的液泡状结构为受精过程中的遗传物质载体; （4） 原胚的形成经历了游离核期和细胞化期。该文针对国内外最新银杏生殖生物学方面的研究进展, 对银杏雌雄配子体发育、受精过程以及胚胎形成等方面进行较为系统全面的分析和总结, 为进一步的银杏生殖生物学研究提供有价值的参考资料。     

Systematic analysis of multigene predictors in gastric cancer exploiting gene expression signature

Gastric cancer (GC) is the second most common cause of cancer death worldwide but could be more curable if diagnosed at an earlier stage. At present, the capability to predict the efficaciousness of molecular diagnosis for GC for each patient remains elusive. The purpose of this study was to identify tumor biomarkers through systems analysis of multigene predictors exploiting the available data resource. In this study, we investigated the top 10% overexpressed genes in GC from five data sets of the Oncomine platform, with 265 GC samples versus 174 normal gastric mucosa samples. Sixteen candidate genes were identified as predictors of GC, of which 14 genes were verified through the comparison of expression levels in specimens from normal (chronic gastritis, 21 samples) and GC groups (38 samples). In addition, unique molecular portraits of diffuse adenocarcinoma (DA), intestinal adenocarcinoma (IA), and mixed adenocarcinoma (MA) were studied through Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway analysis, where DA showed higher extracellular matrix alteration while IA and MA showed higher cell-cycle alteration than other types. We also found that the elevated expressions of genes during GC progression were independent of gene mutations, and high core-binding factor subunit β expression is correlated with a high overall survival rate in GC patients. Our research may provide an efficient clinical diagnosis of GC at an early stage with high accuracy and thus help improve the overall survival rate through early therapeutic interventions.