Benzene containing polyhydroxyalkanoates homo- and copolymers synthesized by genome edited Pseudomonas entomophila

Microbial synthesis of functional polymers has become increasingly important for industrial biotechnology.For the first time,it became possible to synthesize controllable composition of poly(3-hydroxyalkanoate)(P3HA)consisting of 3-hydroxydodecanoate(3HDD)and phenyl group on the side-chain when chromosome of Pseudomonas entomophila was edited to weaken itsβ-oxidation.Cultured in the presence of 5-phenylvaleric acid(PVA),the edited P.entomophila produced only homopolymer poly(3-hydroxy-5-phenylvalerate)or P(3HPhV).While copolyesters P(3HPhV-co-3HDD)of 3-hydroxy-5-phenylvalerate(3HPhV)and 3-hydroxydodecanoate(3HDD)were synthesized when the strain was grown on mixtures of PVA and dodecanoic acid(DDA).Compositions of 3HPhV in P(3HPhV-co-3HDD)were controllable ranging from 3%to 32%depending on DDDA/PVA ratios.Nuclear magnetic resonance(NMR)spectra clearly indicated that the polymers were homopolymer of P(3HPhV)and random copolymers of 3HPhV and 3HDD.Their mechanical and thermal properties varied dramatically depending on the monomer ratios.Our results demonstrated the possibility to produce tailor-made,novel functional PHA using the chromosome edited P.entomophila.     

Combination of Intratumoral Invariant Natural Killer T Cells and Interferon-Gamma Is Associated with Prognosis of Hepatocellular Carcinoma after Curative Resection

Purpose

To investigate the prognostic value of intratumoral invariant natural killer T (iNKT) cells and interferon-gamma (IFN-γ) in hepatocellular carcinoma (HCC) after curative resection.

Experimental Design

Expression of TRAV10, encoding the Vα24 domain of iNKT cells, and IFN-γ mRNA were assessed by quantitative real-time polymerase chain reaction in tumor from 224 HCC patients undergoing curative resection. The prognostic value of these two and other clinicopathologic factors was evaluated.

Results

Either intratumoral iNKT cells and IFN-γ alone or their combination was an independent prognostic factor for OS (P = 0.001) and RFS (P = 0.001) by multivariate Cox proportional hazards analysis. Patients with concurrent low levels of iNKT cells and IFN-γ had a hazard ratio (HR) of 2.784 for OS and 2.673 for RFS. The areas under the curve of iNKT cells, IFN-γand their combination were 0.618 vs 0.608 vs 0.654 for death and 0.591 vs 0.604 vs 0.633 for recurrence respectively by receiver operating characteristic curve analysis. The prognosis was the worst for HCC patients with concurrent low levels of iNKT cells and IFN-γ, which might be related with more advanced pTNM stage and more vascular invasion.

Conclusions

Combination of intratumoral iNKT cells and IFN-γ is a promising independent predictor for recurrence and survival in HCC, which has a better power to predict HCC patients’ outcome compared with intratumoral iNKT cells or IFN-γ alone.     

Diversity and Abundance of Nitrate Assimilation Genes in the Northern South China Sea

Marine heterotrophic microorganisms that assimilate nitrate play an important role in nitrogen and carbon cycling in the water column. The nasA gene, encoding the nitrate assimilation enzyme, was selected as a functional marker to examine the nitrate assimilation community in the South China Sea (SCS). PCR amplification, restriction fragment length polymorphism (RFLP) screening, and phylogenetic analysis of nasA gene sequences were performed to characterize in situ nitrate assimilatory bacteria. Furthermore, the effects of nutrients and other environmental factors on the genetic heterogeneity of nasA fragments from the SCS were evaluated at the surface in three stations, and at two other depths in one of these stations. The diversity indices and rarefaction curves indicated that the nasA gene was more diverse in offshore waters than in the Pearl River estuary. The phylotype rank abundance curve showed an abundant and unique RFLP pattern in all five libraries, indicating that a high diversity but low abundance of nasA existed in the study areas. Phylogenetic analysis of environmental nasA gene sequences further revealed that the nasA gene fragments came from several common aquatic microbial groups, including the Proteobacteria, Cytophaga–Flavobacteria (CF), and Cyanobacteria. In addition to the direct PCR/sequence analysis of environmental samples, we also cultured a number of nitrate assimilatory bacteria isolated from the field. Comparison of nasA genes from these isolates and from the field samples indicated the existence of horizontal nasA gene transfer. Application of real-time quantitative PCR to these nasA genes revealed a great variation in their abundance at different investigation sites and water depths.     

Chinese tomato yelIow Ieaf curl virus— a new species of geminivirus

Chine tomato yellow leaf curl virus (TYLCV-CHI) and other geminiviruses were analysed with 20 monoclonaI antibodies. It was shown that TYLCV-CHI is serclogicaIly close to Chinese tabacco Ieaf curl virus (TbLCV-CHI). The fragment of TYLCV-CHI DNA including the common region (CR), N-terminal of coat protein gene and AV1 gene was amplified by PCR and cloned, and its DNA sequence was determined. These raults showed that TYLCV-CHI is different from other known geminiviruses in the world, and is a new whitefly-transmitted gerninivirus.     

生长特性差别的胃癌MGC803细胞亚系间细胞骨架、细胞通讯与癌基因表达的比较研究

肿瘤细胞表型差异是临床药物敏感性、抗原性、转移性、潜伏性和进展速度等差异的主要原因。为了弄清恶性表型差别的表现、相关性及调节机理、我们从胃癌MGC 803系分离了单细胞亚系,各亚系按克隆过程的时间差别归纳分组,选快组M 17、中组M 6和慢组M 3进行比较。软琼脂生长实验表明三者的恶性程度有显著差别;M17恶性度最高,M3最低,M6居中。与其它表型联系比较,证明细胞生长速度、细胞微丝、微管骨架破坏及细胞通讯功能抑制都与锚着不依赖性生长的恶性特征有平行相关性。同时,M 17和M 3原癌基因表达有明显差别。     

用人工神经网络方法估计Ligistic方程参数

本文提出Logistic方程参数优化估计的人工神经网络方法,并选取一组标样进行具体尝试。结果表明,用这种方法估计Logistic方程参数效果极好。