Hoppel-family of mobile elements of Drosophila melanogaster, flanked by short inverted repeats and having preferential localization in the heterochromatin regions of the genome

A mobile element (ME) having 91% homology with Dm1360 (Kholodilov et al., 1987) has been cloned from the Drosophila melanogaster genome and sequenced. The family of ME was designated hoppel. The members of this family are flanked by short inverted repeats likewise P, hobo and HB. The hoppel is hybridized with 10-30 euchromatic sites of polytene chromosomes of different Drosophila stocks. Abundant hybridization with heterochromatic regions of chromosomes-chromocenter, pericentric heterochromatin, the 4 chromosome and telomeres was observed in all stocks of D. melanogaster examined and in D. simulans. At least six genomic variants of ME differing in length of the central part were revealed. Hoppel possesses ARS activity similar to the P element. Two ME hoppel were shown to be arranged as a direct repeat in the recombinant phage.     

Specificity of the carboxypeptidase inhibitor from potatoes

Carboxypeptidases from animal, plant, fungal, and bacterial sources were tested for their ability to bind to the carboxypeptidase inhibitor from Russet Burbank potatoes. Enzymes which participate in the degradation of dietary protein were partially purified from animal species as diverse as the cow and the limpet, and all were potently affected by the inhibitor. However, several zymogens of the enzymes in this group were tested and shown not to bind immobilized inhibitor. With the exception of an enzyme from mast cells and a novel carboxypeptidase A-like enzyme from bovine placenta, all animal carboxypeptidases which were not of digestive tract origin were not affected by the inhibitor. The inhibitor had no effect on the enzymic activities of all plant and most microbial carboxypeptidases. However, a strong association between the inhibitor and Streptomyces griseus carboxypeptidase has been noted previously and a low affinity (K_i about 10 micromolar) for a carboxypeptidase G₁ from an acinetobacterium was found in this study.     

An improved procedure for production of white spruce (Picea glauca) transgenic plants using Agrobacterium tumefaciens.

An efficient and reproducible procedure for the transformation of white spruce (Picea glauca [Moench] Voss) embryogenic tissues was developed using A. tumefaciens-mediated gene transfer. Rapidly dividing white spruce embryogenic tissues were co-cultivated with disarmed A. tumefaciens strains containing additional copies of the virulence regions from plasmid PToK47. The plasmid pBi121, containing the neomycin phosphotransferase II (nptII) gene providing kanamycin resistance as a selectable marker and the beta-glucuronidase (uidA) reporter gene, was used as binary vector. The highest frequency of transformation (15 transformed tissues g(-1) FW of treated embryogenic tissue) was obtained with 5-d-old tissues grown in liquid medium and co-cultivated with Agrobacterium for 2 d in the same medium but containing 50 microM acetosyringone. Recovery of kanamycin-resistant tissues was improved when tissues were first grown for 10 d on a timentin-containing medium (400 mg l(-1)), to prevent bacterial overgrowth, before application of the selection pressure. After 6 weeks on kanamycin-selection medium, resistant tissues were obtained and showed stable uidA expression. The presence of the transgenes was demonstrated by PCR analysis and their integration into the genome was confirmed by Southern hybridization. Transgenic plants were regenerated from transformed tissues within 4 months after co-culture.     

反例微视频教学法在“微生物学实验”课程中的实践研究

"微生物学实验"是生物类相关专业的一门必修课程,该课程的开展对提高学生实验技能和理解微生物学知识原理至关重要。因此,不断探索和改进实验课程教学方法、减少学生的错误实验操作、提高实验课堂教学效率是任课教师的基本任务。反例微视频教学法是将传统的案例教学法、反例教学法和当下流行的微课程相结合的一种教学方法。本文阐述了该教学法的原理、基本操作流程、课堂应用效果和存在的问题。该教学法的实施能较好地提高实验课堂效率,大幅减少学生错误实验操作的次数,值得在本科生物类实验课堂中进行推广。     

Evolution of heat‐shock protein expression underlying adaptive responses to environmental stress

Heat‐shock proteins (Hsps) and their cognates are primary mitigators of cell stress. With increasingly severe impacts of climate change and other human modifications of the biosphere, the ability of the heat‐shock system to affect evolutionary fitness in environments outside the laboratory and to evolve in response is topic of growing importance. Since the last major reviews, several advances have occurred. First, demonstrations of the heat‐shock response outside the laboratory now include many additional taxa and environments. Many of these demonstrations are only correlative, however. More importantly, technical advances in “omic” quantification of nucleic acids and proteins, genomewide association analysis, and manipulation of genes and their expression have enabled the field to move beyond correlation. Several consequent advances are already evident: The pathway from heat‐shock gene expression to stress tolerance in nature can be extremely complex, mediated through multiple biological processes and systems, and even multiple species. The underlying genes are more numerous, diverse and variable than previously appreciated, especially with respect to their regulatory variation and epigenetic changes. The impacts and limitations (e.g., due to trade‐offs) of natural selection on these genes have become more obvious and better established. At last, as evolutionary capacitors, Hsps may have distinctive impacts on the evolution of other genes and ecological consequences.     

Downregulation of microRNA‐15a‐3p is correlated with clinical outcome and negatively regulates cancer proliferation and migration in human osteosarcoma

In this study, we investigated the expression, correlation to clinical outcomes and biological functions of microRNA‐15a‐3p (miR‐15a‐3p) in human osteosarcoma. MiR‐15a‐3p expressions in osteosarcoma cell lines and clinical tissues of osteosarcoma patients were measured by qPCR. Relevance of endogenous miR‐15a‐3p to osteosarcoma patients' clinicopathological factors or overall survival was statistically analyzed. In addition, the independence of miR‐15a‐3p predicting cancer patients' overall survival was analyzed by Cox regression method. Furthermore, in osteosarcoma cell lines, Saos‐2 and HOS cells, miR‐15a‐3p was overexpressed through stable lentiviral transduction. The functional regulations of miR‐15a‐3p overexpression on cancer ell proliferation and migration were then analyzed. MiR‐15a‐3p was significantly downregulated in osteosarcoma cell lines and human osteosarcoma tumors. Downregulation of endogenous miR‐15a‐3p in osteosarcoma tumors was significantly associated with cancer patient's poor clinical outcomes and low survival rate. Also, endogenous miR‐15a‐3p was confirmed to be an independent biomarker for predicating cancer patients' survival. In Saos‐2 and HOS cells, lentivirus‐induced miR‐15a‐3p overexpression had significantly tumor suppressing functions, by inhibiting both proliferation and migration. Significant downregulation of miR‐15a‐3p in osteosarcoma may be an independent biomarker to predicting cancer patients' poor prognosis. Overexpression miR‐15a‐3p may be an efficient functional meaning to suppress osteosarcoma development.     

In vitro culture expansion impairs chondrogenic differentiation and the therapeutic effect of mesenchymal stem cells by regulating the unfolded protein response

In vitro expansion of mesenchymal stem cells (MSCs) has been implicated in loss of multipotency, leading to impaired chondrogenic potential and an eventual therapeutic effect, as reported in our previous study. However, the precise regulatory mechanism is still unclear. Here, we demonstrate that endoplasmic reticulum (ER) stress and the unfolded protein response (UPR) were involved in transformation of MSCs induced by in vitro culture based on the comparative profiling of in vitro cultured bone marrow MSCs at passage 3 (P3 BMSCs) vs. fresh P0 BMSCs by microarray analysis. Indeed, RT-PCR and Western blot analysis showed significantly lower expression levels of three key UPR-related molecules, ATF4, ATF6 and XBP1, in P3 BMSCs than P0 BMSCs. Further, we found that UPR suppression by 4-phenylbutyrate (4-PBA) reduced the chondrogenic potential of P0 BMSCs and further cartilage regeneration. Conversely, UPR induction by tunicamycin (TM) enhanced the chondrogenic differentiation of P3 BMSCs and the therapeutic effect on cartilage repair. Thus, the decline in the chondrogenic potential of stem cells after in vitro culture and expansion may be due to changes in ER stress and the UPR pathway.     

氟西汀联合同步放化疗治疗局部晚期非小细胞肺癌的临床观察

目的：观察氟西汀联合同步放化疗治疗局部晚期非小细胞肺癌（none small cells lung carcinoma,NSCLC）的临床疗效及其安全性。方法：选择2010年10月～2011年4月我院收治的III期不能手术的NSCLC患者47例,根据患者的入院顺序,随机分为两组,同步放化疗组（对照组）22例,接受三维适形或调强放疗：肿瘤处方剂量60~66Gy,常规分割：1．8～2．0Gy／次;放疗期间同步两周期“紫杉醇＋顺铂”方案化疗,放疗后再原方案巩固化疗2--3个周期;氟西汀联合治疗组（实验组）25例,从同步放化疗开始起同时服用氟西汀（20-40mg／day）,持续服药半年。随访至1年,观察和评价两组的疗效以及不良反应的发生情况。结果：对照组与实验组的客观有效率分别为77-3％（17／22）和80％（20／25）（P〉0．05）;1年生存率分别为68．2％（15／22）和80％（20／25）（P〉0．05）;1年局控率分别为45．5％（10／22）和76％（19／25）,差异有统计学意义（P〈0．05）。治疗后,实验组患者CD＋／CD8＋比率由1．34±0．23升至1．58±0．22（P〈0．05）,而汉密尔顿抑郁量表（HAMD）总分由13．4±4．8降至9．6±3（P〈0．05）;全组患者主要不良反应为骨髓抑制、消化道反应、放射性食管炎和放射性肺炎,来发生4级不良反应,两组3级不良反应发生率无明显差异（P〉0．05）。结论：对于不能手术的局部晚期NSCLC患者,在同步放化疗的基础上联合氟西汀治疗可以提高肿瘤的1年局控率,促进抗肿瘤免疫,并缓解患者的抑郁情绪,且不加重不良反应。