Identification of the gene imds-60 in mouse epididymis

Sperm maturation, including the acquisition of motility and the full ability to fertilize oocyte, occurs during its transit through the dynamic environment of the epididymis. However, the roles of many genes involved in the process of sperm maturation still remain to be found. Based on an expressed sequence tag named imds-60, which was first found in uterus but is highly expressed in epididymis, the full-length cDNA sequence of imds-60 with a complete open reading frame was obtained in mouse epididymis by GenBank searching, polymerase chain reaction-based procedures, and 5＇- and 3＇-rapid amplification of cDNA ends. This protein was predicted to have an N-terminal signal peptide and a C-terminal DNase I-like domain with nine transmembrane motifs in the middle part of the protein. Northern blot analysis showed that the mRNA of imds-60 was highly expressed in epididymis but at a rather lower level in uterus, seminal vesicle gland, and stomach. Further study revealed that the mRNA of imds-60 is only expressed in corpus and cauda regions of epididymis, not in caput. It is regulated partially by androgen and peaked in male mice aged from 3 weeks to adult. The imds-60 protein might play an important role in cell communication during sperm maturation.     

虫草多糖治疗家犬夹闭肾动脉引起的急性肾衰

探讨了虫草多糖对家犬夹闭肾动脉造成的急性肾衰的治疗作用。将实验动物家犬分为假手术组、阴性模型组、阳性组、虫草多糖高、中、低三个剂量组,观察治疗后犬的肾功能、血液生化指标、尿常规、肾脏病理的变化。发现虫草多糖组肾功能受损程度明显轻于阴性模型组（P〈0．01）,肾脏病理损伤明显减轻。实验证明虫草多糖能够治疗肾缺血诱发的急性肾功能衰竭,其机理可能与改善肾功能、增加肾血流量、纠正代谢紊乱、促进肾小管的修复与再生等作用有关。     

高效液相色谱法测定大黄素在Caco-2细胞中的摄取

目的:采用反相高效液相色谱法,观察大黄素在Caco-2细胞中的摄取特点。方法:将大黄素与Caco-2细胞共同孵育,收集细胞样品,液氮反复冻融。取细胞裂解液,加入甲醇提取,提取液采用HPLC进行分析。色谱分析柱为C18柱(250mm×4．6mm,5μm,Diamonsil),流动相组成为85%乙腈及15%水(含0．1%乙酸),流速1ml·min-1,进样量20μl,柱温25℃,3D模式采集数据。结果:检测Caco-2细胞中大黄素的工作曲线的回归方程为Y=0．278x 0．148(Y=0．9996,n=5),线性范围为0．037～4．8μmol·L-1,最低检测浓度为0．018μmol·L-1。当细胞中大黄素的浓度为0．05、2和8．5μg·ml-1时,回收率分别为(101．3±7．3)%、(96．7±3．0)%和(98．7±2．1)%(n=5);相应的日内标准偏差分别为0．25%、2．9%和1．4%;相应的日间标准偏差分别为2．3%、5．6%和6．3%。大黄素在Caco-2细胞中的摄取达峰时间为10分钟,峰浓度为108．56±11．57 nmol/L·mg·protein,10分钟后Caco-2细胞中大黄素的含量迅速下降。浓度处于2-50μM之间时,Caco-2细胞对大黄素的摄取量呈线性增加,浓度达50μM后,随着剂量的增加大黄素的摄取量变化不明显。结论:大黄素可被Caco-2细胞迅速摄取,随着剂量的增加,大黄素在Caco-2细胞中的摄取存在饱和现象。     

籼稻稻米蛋白质含量与外观品质性状间不同遗传体系的条件遗传相关分析

选用稻米蛋白质含量等品质差异较大的7个籼型不育系(A)及相应的保持系(B)与5个籼型恢复系(R)杂交,组成7×5不完全双列杂交组合。应用包括胚乳、细胞质和母体植株基因的遗传主效应以及基因型×环境互作效应的数量性状遗传模型及非条件和条件分析方法,研究了籼稻稻米蛋白质含量与外观品质性状间的遗传相关性,进一步揭示了籼稻糙米重、稻米直链淀粉含量对稻米蛋白质含量与外观品质性状间遗传相关性的影响。非条件分析的结果表明,除了与糙米厚的相关性未达到显著水平以外,蛋白质含量与其它稻米外观品质性状间的遗传相关性均达极显著水平,其中与糙米宽间的相关性表现为正值,其余为负向相关。条件分析的结果显示,糙米重和直链淀粉含量对稻米蛋白质含量与外观品质性状间的遗传相关性均可产生较大的影响。糙米重主要通过基因型×环境互作效应影响蛋白质含量与外观品质性状间的遗传相关性,其中对蛋白质含量与糙米长、糙米宽、糙米厚间遗传相关性的影响主要表现为负向作用,而对蛋白质含量与糙米长宽比、糙米长厚比间遗传相关性的影响则表现为正向作用。稻米直链淀粉主要通过细胞质遗传主效应和母体加性效应影响蛋白质含量与糙米长、糙米长宽比、糙米长厚比间的遗传相关性,而对蛋白质含量与糙米宽间的相关性影响主要表现为负向作用。     

Breeding and application of Jiafuzhan, a new elite early indica rice cultivar in China

After 20 years of dedicated research,Jiafuzhan has been successfully developed under the new technologies in breeding high-quality early indica rice cultivars.Its rice quality has almost reached the A-level Editable Rice of Agriculture Department of China,and its average production reaches 400-500 kg/(666.7 m2).This new cultivar also has other characteristics such as enhanced resistance of blast and fallen,steady productivity,and strong adaptability.Jiafuzhan has been put into production of over 11.4×104 hm2 in Fujian Province and has been introduced and extended in other Provinces like Jiangxi,Guangdong,and Guangxi,China.The successes of breeding Jiafuzhan is a solution to the existing perennial problems in the rice industry,such as poor grain quality of big-grain rice and early indica rice,low productivity,and poor blast resistance of elite rice.     

The retention properties of nucleobases in alkyl C8-/C18- and IAM-chromatographic systems in relation to log Pow

In order to evaluate the differences in the partition properties of 35 structurally congeneric nucleobases of biological interests in octanol-water biphasic, alkyl C(8)/C(18), and IAM systems, a comparative chromatographic study was performed. Comparing with the reversed-phase C(8)/C(18) retention data, most of the purines possessed weaker IAM retention except for those with specific H-bond and/or electrostatic interactions. Quantitative correlations between the experimental log P(ow) literature values and the IAM, C(8), and C(18) log k were evaluated (R(2)=0.943, 0.794, and 0.767, respectively). Although IAM retention correlated significantly better (larger R(2) value) with the log P(ow) values statistically, the latter was revealed apparently behaving more like (slope approaching unity) alkyl C(8)/C(18) retention and hence also has the same shortcoming in under-representing analytes capable of forming short-term H-bond/electrostatic interactions with polar head-groups of phospholipids. A chemically meaningful structure-retention model (q(2)=0.824 and R(2)=0.968) was derived, in which the hydrophobic interaction is identified as the underlying factor for the retention of purines in IAM system modulated non-trivially by H-bond/electrostatic interactions.     

Molecularly imprinted polymer microspheres for solid-phase extraction of chloramphenicol residues in foods

Preparation of molecularly imprinted polymer microspheres (MIPMs) for chloramphenicol (CAP) by aqueous suspension polymerization is reported for the first time in this study. The resulting MIPMs had the ability to specifically adsorb CAP, and the molecularly imprinted solid phase extraction (MISPE) based on the MIPMs was shown to be applicable for clean-up and preconcentration of trace CAP in milk and shrimp samples with high recoveries of 92.7% and 84.9%, respectively. Combined with MISPE, the conventional HPLC-UV analysis sensitivity for CAP in foods could be significantly increased.     

Quantitative determination of trans-polydatin, a natural strong anti-oxidative compound, in rat plasma and cellular environment of a human colon adenocarcinoma cell line for pharmacokinetic studies

A simple, accurate, precise, specific and reproducible high-performance liquid chromatography (HPLC) method was developed for determination of trans-polydatin, a natural strong anti-oxidative compound, in rat plasma and cell suspension. The assay procedure involved simple liquid-liquid extraction, the supernatant liquid was added an equal volume of water to avoid solvent effect. The detection of the analyte peak was achieved by monitoring the eluate using a UV detector set at 303 nm. The analysis used a Hypersil ODS2 C18 column (5 microm, 4.6 mm x 250 mm) and methanol/distilled water as the mobile phase (flow rate=1 mL/min). A total analytical run was achieved within 6.0 min and calibration curve was linear over a wide concentration range of 0.25-40 microg/mL for plasma sample and 1.0-500 microM for cell suspension, the coefficients of correlation were 0.9997 and 0.9999 or better, respectively. There was 80.7+/-7.86%, 96.8+/-3.20% and 102.7+/-9.72% recovery from 0.5, 10, and 40 microg/mL plasma samples, respectively. Intra- and inter-batch accuracy and precision were acceptable for the both matrices. The RSD of intra- and inter-day assay variations were all less than 10%. Both analyte and IS were stable in the battery of stability studies, freeze-thaw cycles. The described assay method was applied to pharmacokinetic studies in rats and a human colon adenocarcinoma cell line (Caco-2) successfully. The application of the assay to determine the pharmacokinetic is described.     

Membrane subproteomic analysis of Mycobacterium bovis bacillus Calmette-Guérin

Mycobacterium bovis bacillus Calmette-Guérin (BCG) vaccine has been known for a long time to prevent tuberculosis (TB) worldwide since 1921. Nonetheless, we know little about BCG membrane proteome. In the present study, we utilized alkaline incubation and Triton X-114-based methods to enrich BCG membrane proteins and subsequently digested them using proteolytic enzyme. The recovered peptides were further separated by 2-D LC and identified by ESI-MS/MS. As a result, total 474 proteins were identified, including 78 integral membrane proteins (IMPs). Notably, 18 BCG IMPs were described for the first time in mycobacterium. Further analysis of the 78 IMPs indicated that the theoretical molecular mass distribution of them ranged from 8.06 to 167.86 kDa and pI scores ranged from 4.40 to 11.60. Functional classification revealed that a large proportion of the identified IMPs (67.9%, 53 out of 78) were involved in cell wall and cell processes functional group. In conclusion, here we reported a comprehensive profile of the BCG membrane subproteome. The present investigation may allow the identification of some valuable vaccine and drug target candidates and thus provide basement for future designing of preventive, diagnostic, and therapeutic strategies against TB.     

In situ proteolysis for protein crystallization and structure determination

We tested the general applicability of in situ proteolysis to form protein crystals suitable for structure determination by adding a protease (chymotrypsin or trypsin) digestion step to crystallization trials of 55 bacterial and 14 human proteins that had proven recalcitrant to our best efforts at crystallization or structure determination. This is a work in progress; so far we determined structures of 9 bacterial proteins and the human aminoimidazole ribonucleotide synthetase (AIRS) domain.