Expression of Recombinant Carcinoembryonic-Antigen-Human-β-Defensin-2 Gene in Colon Cancer HCT116 Cells

The objective of this study was to construct a system driving high expression of human-β-denfensin-2 (HBD-2) system in eukaryotic cell. To construct recombinant retrovirus expression vector, CEA signal peptide-HBD-2 (mature peptide sequence) was cloned in the retrovirus expression vector PLNCX2. The retroviral vector was transfected into PT67 cells by DOTAP; after screening and amplifying single clone cell by G418, the virus particles were collected and infected to eukaryotic, colon cancer HCT116 cells. Furthermore, the HCT116 cells were also screened by using G418 and the resistance clones were obtained. Finally, the expression of HBD-2 was detected by Western blotting, which suggested a high level of expression of HBD-2 in HCT116 cells. In conclusion, the results indicate that high level of HBD-2 expression was obtained in HCT116 cells which will help in effective commercial production and purification of HBD-2 protein.     

Long-Term Outcome of Sensorineural Hearing Loss in Nasopharyngeal Carcinoma Patients: Comparison Between Treatment with Radiotherapy Alone and Chemoradiotherapy

The purpose of this study is to assess the long-term effect of sensorineural hearing loss (SNHL) resulted from radiotherapy (RT) alone versus chemoradiotherapy in nasopharyngeal carcinoma patients (NPC). Seventy-two patients initially diagnosed with NPC were enrolled from Shandong Tumor Hospital between March 2003 and May 2007. They were assigned into two groups: RT alone and chemoradiotherapy according to the different treatment regimens. Intensity-modulated radiation therapy was applied for both groups, concurrent and adjuvant cisplatin were administered for chemoradiotherapy group additionally. Hearing threshold test was performed at various time periods after completion of RT. Mean radiation dose to the cochlea in each ear was calculated to determine the correlation between cochlear dose and SNHL. We found that the hearing loss is more severe in the chemoradiotherapy group compared with RT group, from completion of RT up to the 5 years of follow-up period. This is especially obvious in the high frequency range. Hearing level is seriously damaged when cochlea dose exceeds 46 GY. We concluded that concurrent/adjuvant chemotherapy plus RT aggravates SNHL in NPC patients than RT alone and thus inner ear tissue tolerance should be redefined in those patients.     

抗体治疗的重生

近年来,治疗性单克隆抗体已成为基础和临床医学研究者及企业关注的热点。目前,针对免疫检查点的治疗性抗体用于肿瘤治疗已显示出较好疗效。在微生物耐药性日益增多、全球突发传染病威胁依然存在及持续性微生物感染难以治愈的当下,抗微生物领域中的抗体治疗正在积极研发中。本文综述了抗体治疗在抗微生物感染领域中的进展,并展望了其应用前景。     

Genomewide identification of PPR gene family and prediction analysis on restorer gene in <Emphasis Type="BoldItalic">Gossypium</Emphasis>

Pentatricopeptide repeat (PPR) gene family plays an essential role in the regulation of plant growth and organelle gene expression. Some PPR genes are related to fertility restoration in plant, but there is no detailed information in Gossypium. In the present study, we identified 482 and 433 PPR homologues in Gossypium raimondii (\(\hbox {D}_{5}\)) and G. arboreum (\(\hbox {A}_{2}\)) genomes, respectively. Most PPR homologues showed an even distribution on the whole chromosomes. Given an evolutionary analysis to PPR genes from G. raimondii (\(\hbox {D}_{5}\)), G. arboreum (\(\hbox {A}_{2}\)) and G. hirsutum genomes, eight PPR genes were clustered together with restoring genes of other species. Most cotton PPR genes were qualified with no intron, high proportion of \(\upalpha \)-helix and classical tertiary structure of PPR protein. Based on bioinformatics analyses, eight PPR genes were targeted in mitochondrion, encoding typical P subfamily protein with protein binding activity and organelle RNA metabolism in function. Further verified by RNA-seq and quantitative real-time PCR (qRT-PCR) analyses, two PPR candidate genes, Gorai.005G0470 (\(\hbox {D}_{5}\)) and Cotton_A_08373 (\(\hbox {A}_{2}\)), were upregulated in fertile line than sterile line. These results reveal new insights into PPR gene evolution in Gossypium.     

Treatment of donor cells with recombinant KDM4D protein improves preimplantation development of cloned ovine embryos

Incomplete epigenetic reprogramming is one of the major factors affecting the development of embryos cloned by somatic cell nuclear transfer (SCNT). Histone 3 lysine 9 (H3K9) trimethylation has been identified as a key barrier to efficient reprogramming by SCNT. The aim of this study was to explore a method of downregulating H3K9me3 levels in donor cells by using histone lysine demethylase (KDM) protein. When sheep fetal fibroblast cells were treated with recombinant human KDM4D protein (rhKDM4D), the levels of H3K9 trimethylation and dimethylation were both significantly decreased. After SCNT, rhKDM4D-treated donor cells supported significantly higher percentage of cloned embryos developing into blastocysts as compared to non-treated control cells. Moreover, the blastocyst quality was also improved by rhKDM4D treatment of donor cells, as assessed by the total cell number in blastocysts and the expression of developmental genes including SOX2, NANOG and CDX2. These results indicate that treatment of donor cells with recombinant KDM4D protein can downregulate the levels of H3K9 trimethylation and dimethylation and improve the developmental competence of SCNT embryos. This strategy may be convenient to be used in KDM4-assisted SCNT procedure for improving the efficiency of cloning.     

糖化酶基因的克隆和在酿酒酵母中表达

以穿梭质粒pLCl为载体,大肠杆菌C600为受体构建了扣囊拟内孢霉(Endomycopsis fibuligera)Sau34基因文库。从基因文库中提取重组质粒DNA,转化酿酒酵母BJ1991,选出具有淀粉水解酶活性的转化子,它含有编码扣囊拟内孢霉胞外糖化酶的基因片段,能发酵淀粉。琼脂糖凝胶电泳证明所插入DNA片段为5.3kb,SDS-聚丙烯酰胺凝胶电泳测得糖化酶的分子量为54000。酶活最适温度为50℃,最适pH为5.5。     

Observation and modeling of plant biomass of meadow steppe in Tumugi, Xingan League, Inner Mongolia, China

We examined the long-term dynamics of plant biomass of Filifolium sibiricum steppe, Stipa baicalensis steppe and Leymus chinense (syn. Aneurolepidium chinense) steppe relative to interannual variation of precipitation and temperature during 1981–1990 in the Tumugi, Xingan League, eastern Inner Mongolia of China. On the average, annual precipitation was 41.1 cm. Peak live aboveground biomass (PLAB) was 152 g m^-2 for F. sibiricum steppe and S. baicalensis steppe and 162 g m^-2 for L. chinense steppe. Peak live belowground biomass (PLBB) varied between 968 g m^-2 for F. sibiricum steppe and 1022 g m^-2 for L. chinense steppe. The coefficient of variation (CV) in annual precipitation (25%) was lower than the CV in PLAB (35% to 37%) but larger than the CV in PLBB (10%) of the three meadow steppe sites. Rain use efficiency was 3.6 gDM m^-2 cm^-1 yr^-1 for F. sibiricum steppe and S. baicalensis steppe, and 3.9 gDM m^-2 cm^-1 yr^-1 for L. chinense steppe, respectively.Using the CENTURY ecosystem model, simulation results agreed reasonably well with the observed soil organic matter, seasonal dynamics and interannual variation of plant biomass of these three steppe sites during 1981–1990. The CENTURY model is slightly more successful than the empirical regression models that use annual precipitation to estimate PLAB of these meadow steppe over time. Both seasonal distribution and interannal variation in precipitation and temperature are the important controls of temporal dynamics of plant biomass, rain use efficiency, carbon flux and storage of these meadow steppe ecosystems over time.