人载脂蛋白E7转基因小鼠肾脏cDNA削减文库与cDNA文库

目的　以人apoE7基因制备的高脂血症转基因小鼠模型已建成 ,要用于探查这一突变基因致病的分子机制 ,首先揭示人apoE7在小鼠体内诱发了那些基因的表达异常 ,是重要的环节之一。cDNA削减文库是寻找异常高表达的已知和未知基因有效的手段 ,而cDNA文库更是获取全长基因所必须。方法　自正常小鼠与转基因小鼠肾脏同时提取总RNA ,再分离mRNA ,逆转录制备cDNA后 ,用两次分子杂交削除转基因鼠cDNA中与正常小鼠相同的部分 ,再用PCR与择需PCR法扩增转基因鼠特异的基因 ,克隆入pGEM T载体后 ,制备削减文库。未经削减的转基因小鼠cDNA克隆入λgt11载体 ,制备基因表达文库。结果和结论　自高脂血症转基因小鼠肾脏构建的削减文库得到约 4 0 0个削减克隆 ,测定了部分克隆的序列 ,并进行同源性比较。λgt11 cDNA文库滴度 1 7× 10 7pfu ml,随机测定的克隆其插入片段长约 5 0 0bp以上。     

中国真叶蝇属六新种及中国种类检索表（双翅目：叶蝇科）

记述中国真叶蝇属6新种:短须真叶蝇P.brevipalpis sp.nov.,瘤突真叶蝇P.gangliiformisa sp.nov.,李氏真叶蝇P.lii sp.nov.,长毛真叶蝇P.longisetae sp.nov.,黑腹真叶蝇P.melanogastera sp.nov.和黑缘真叶蝇P.nigrimarginata sp.nov.,编制了本属中国种类的检索表.     

Alterations in the gut microbiota of patients with acquired immune deficiency syndrome

Acquired immune deficiency syndrome (AIDS), caused by infection with human immunodeficiency virus (HIV), is associated with gastrointestinal disease, systemic immune activation and changes in the gut microbiota. Here, we aim to investigate the gut microbiota patterns of HIV‐infected individuals and HIV‐uninfected individuals in populations from South China. We enrolled 33 patients with HIV (14 participants treated with highly active antiretroviral therapy [HAART] for more than 3 months; the remaining 19 individuals had not received treatment) and 35 healthy controls (HC) for a cross‐sectional comparison of gut microbiota using stool samples. Gut microbial communities were profiled by sequencing the bacterial 16S rRNA genes. Dysbiosis was more common among patients with AIDS compared with healthy individuals. Dysbiosis was characterized by decreased α‐diversity, low mean counts of Bacteroidetes, Faecalibacterium, Prevotella, Bacteroides vulgatus, Dialister and Roseburia inulnivorans, and high mean counts of Proteobacteria, Enterococcus, Streptococcus, Lactobacillus, Lachnociostridium, Ruminococcus gnavus and Streptococcus vestibularis. Increased abundance of Bacilli was observed in homosexual patients. Proteobacteria were higher among heterosexual patients with HIV infections. Tenericutes were higher among patients with history of intravenous drug abuse. Restoration of gut microbiota diversity and a significant increase in abundance of Faecalibacterium, Blautia and Bacteroides were found in patients receiving HAART compared to those who did not receive. HIV infection‐associated dysbiosis is characterized by decreased levels of α‐diversity and Bacteroidetes, increased levels of Proteobacteria and the alterations of gut microbiota correlate with the route of HIV transmission. The imbalanced faecal microbiota of HIV infection is partially restored after therapy.     

Albumin's Influence on Carprofen Enantiomers–Hymecromone Interaction

Hymecromone is an important coumarin drug, and carprofen is one of the most important nonsteroidal antiinflammatory drugs (NSAIDs). The present study aims to determine the influence of bovine serum albumin (BSA) on the carprofen–hymecromone interaction. The inhibition of carprofen enantiomers on the UDP‐glucuronosyltransferase (UGT) 2B7‐catalyzed glucuronidation of hymecromone was investigated in the UGTs incubation system with and without BSA. The inhibition capability of increased by 20% (P < 0.001) of (R)‐carprofen after the addition of 0.5% BSA in the incubation mixture. In contrast, no significant difference was observed for the inhibition of (S)‐carprofen on UGT2B7 activity in the absence or presence of 0.5% BSA in the incubation system. The Lineweaver‐Burk plot showed that the intersection point was located in the vertical axis, indicating the competitive inhibition of (R)‐carprofen on UGT2B7 in the incubation system with BSA, which is consistent with the inhibition kinetic type of (R)‐carprofen on UGT2B7 in the incubation system without BSA. Furthermore, the second plot using the slopes from the Lineweaver‐Burk versus the concentrations of (R)‐carprofen showed that the fitting equation was y=39.997x+50. Using this equation, the inhibition kinetic parameter was calculated to be 1.3 μM. For (S)‐carprofen, the intersection point was located in the horizontal axis in the Lineweaver‐Burk plot for the incubation system with BSA, indicating the noncompetitive inhibition of (S)‐carprofen on the activity of UGT2B7. The fitting plot of the second plot was y=24.6x+180, and the inhibition kinetic parameter was 7.3 μM. In conclusion, the present study gives a short summary of BSA's influence on the carprofen enantiomers–hymecromone interaction, which will guide the clinical application of carprofen and hymecromone. Chirality 28:226–229, 2016. © 2015 Wiley Periodicals, Inc.     

Tetramethylpyrazine Nitrone Improves Neurobehavioral Functions and Confers Neuroprotection on Rats with Traumatic Brain Injury

Oxidative stress is one of the major secondary injury mechanisms after traumatic brain injury (TBI). 2-[[(1,1-Dimethylethyl)oxidoimino]-methyl]-3,5,6-trimethylpyrazine (TBN), a derivative of the clinically used anti-stroke drug tetramethylpyrazine armed with a powerful free radical-scavenging nitrone moiety, has been demonstrated promising therapeutic efficacy in ischemic stroke and Parkinson’s models. The present study aims to investigate the effects of TBN on behavioral function and neuroprotection in rats subjected to TBI. TBN (90 mg/kg) was administered twice daily for 7 days by intravenous injection following TBI. TBN improved neuronal behavior functions after brain injury, including rotarod test and adhesive paper removal test. Compared with the TBI model group, TBN treatment significantly protected NeuN-positive neurons, while decreased glial fibrillary acidic protein (GFAP)-positive cells. The number of 4-hydroxynonenal (4-HNE)-positive and 8-hydroxy-2′-deoxyguanosine (8-OHdG)-positive cells around the damaged area after TBI were significantly decreased in the TBN treatment group. In addition, TBN effectively reversed the altered expression of Bcl-2, Bax and caspase 3, and the down-regulation of nuclear factor erythroid-derived 2-like 2 (Nrf-2) and hemeoxygenase-1 (HO-1) proteins expression stimulated by TBI. In conclusion, TBN improves neurobehavioral functions and protects neurons against TBI. This protective effect may be achieved by anti-neuronal apoptosis, alleviating oxidative stress damage and up-regulating Nrf-2 and HO-1 expression.     

Somatic embryogenesis and plant regeneration from different wild diploid cotton (Gossypium) species

Calli were successfully induced from hypocotyls of eight wild diploid cotton species (Gossypium) on MSB (MS salts and B₅ vitamins) medium supplemented with 0.09 μM 2,4-D (2,4-dichlorophenoxyacetic acid) and 2.32 μM KT (kinetin). Plant growth regulator (PGR) combinations, adding GA₃ (Gibberellic acid), high inorganic salt stress, and PGR-free media were used to induce embryogenic calli from nonembryogenic calli. Embryogenic cultures were induced from G. aridum S. (D₄ genome), G. davidsonii K. (D₃-d genome), G. klotzschianum A. (D₃-k genome), G. raimondii U. (D₅ genome), and G. stocksii M. (E₁ genome). We then observed somatic embryogenesis in the five species while calli of G. africanum V. (A₁-2 genome), G. anomalum W. (B₁ genome), and G. bickii P. (G genome) remained nonembryogenic. Somatic embryogenesis was adjusted by changing sugar sources, regulating combinations of PGRs, and using cell suspension culture. Embryos at various developmental stages produced mature and germinating embryos when cultured on filter paper placed on the media containing different sugar sources. The utility of different sugar sources promoted globular embryos developing into cotyledonary stage and increased the frequency of cotyledonary embryos developing into normal plants. Normal plantlets were regenerated from G. davidsonii, G. klotzschianum, G. raimondii, and G. stocksii. Only abnormal plantlets were obtained in G. aridum. This work will contribute to broadening the number of regenerable cotton species and provide foundations for somatic hybridization in cotton to create new germplasm.     

Polo-like kinase 3 is Golgi localized and involved in regulating Golgi fragmentation during the cell cycle

The Golgi apparatus undergoes extensive fragmentation during mitosis in animal cells. Protein kinases play a critical role during fragmentation of the Golgi apparatus. We reported here that Polo-like kinase 3 (Plk3) may be an important mediator during Golgi breakdown. Specifically, Plk3 was concentrated at the Golgi apparatus in HeLa and A549 cells during interphase. At the onset of mitosis, Plk3 signals disintegrated and redistributed in a manner similar to those of Golgi stacks. Nocodazole activated Plk3 kinase activity, correlating with redistribution of Plk3 signals and Golgi fragmentation. In addition, treatment with brefeldin A (BFA), a Golgi-specific poison, also resulted in disappearance of concentrated Plk3 signals. Plk3 interacted with giantin, a Golgi-specific protein. Expression of Plk3, but not the kinase-defective Plk3 (Plk3(K52R)), resulted in significant Golgi breakdown. Given its role in cell cycle progression, Plk3 may be a protein kinase involved in regulation of Golgi fragmentation during the cell cycle.     

Somatic embryogenesis and plant regeneration in wild cotton (Gossypium klotzschianum)

A simple and efficient method for high frequency somatic embryogenesis and plant regeneration from hypocotyl-derived cultures and suspension cultures of Gossypium klotzschianum Anderss, a wild, diploid species of cotton is described here. Embryogenic cultures were induced from hypocotyl sections on MSB medium with 0.9 M 2,4-D and 2.32 M kinetin. MSB medium containing 0.045 M 2,4-D, 0.93 M kinetin, 2.46 M IBA promoted embryogenic culture proliferation and embryo development. Suspension cultures with 0.23 M 2,4-D and 0.93 M kinetin also produced many embryos. Somatic embryos cultured on MSB medium with PGRs produced secondary embryos, and embryos developed into normal plantlets on PGR-free MSB medium. Regenerated plantlets were transferred onto the quarter-strength MSB medium with 0.5% active charcoal to avoid recallusing. Hypocotyls were better than cotyledons for culture induction and plant regeneration. 2,4-D and kinetin were essential for culture induction and maintenance.