Biosynthesis of proteoglycans in organ cultures of developing kidney mesenchyme

The biosynthesis of proteoglycans was studied in organ cultures of differentiating metanephric mesenchymes. When triggered by a contact-mediated inductive interaction, this tissue undergoes transition from a mesenchyme to an epithelium. In the present study, proteoglycans were extracted by guanidinium hydrochloride in the presence of protease inhibitors. We found that, as a response to induction, the differentiating mesenchyme begins to synthesize large size proteoglycans with an apparent molecular weight (MW) of 1 X 10(6) D. The major glycosaminoglycans detected were chondroitin sulfates. Heparan sulfate proteoglycans were also detected, constituting 20% of the proteoglycans. An inhibitor of glucosamine synthesis, 6-diazo-5-oxo-norleucine (DON) was found to inhibit glycosaminoglycan synthesis by approx. 60%, and the size of the proteoglycans was also diminished. Our studies suggest that the transition of the mesenchyme to epithelium is associated with initiation of synthesis of large size proteoglycans.     

Formation of copper-bleomycin complexes: evidence of a three-step process

The formation of Cu(II)-bleomycin complexes as a function of pH has been studied using circular dichroism, absorption, electron paramagnetic resonance spectroscopy, and potentiometric titration. Our data support the following points: the formation of Cu(II)-bleomycin complexes occurs in a three-step process: a first complex (I) is formed at pH 1.2, which most probably involves the pyrimidine nitrogen, the secondary amine nitrogen, and two water molecules as the four in-plane ligands of copper. A second complex (II) is formed at pH 2.5, through the further coordination of the peptide nitrogen of histidine residue, and histidine imidazole nitrogen giving rise to the release of two protons. The fixation, in apical position, of the alpha-amino nitrogen of beta-aminoalanine occurs in a last step through the release of one additional proton. A value of 2.7 has been obtained for the pK of formation of this third complex, which is the species present at physiological pH. In the Cu(II)-depbleomycin system only one complex (II') has been detected.     

接种不同AM真菌对滇重楼幼苗功能基因表达的影响

为探究接种不同外源性从枝菌根(Arbuscular mycorrhizae,AM)真菌对滇重楼幼苗基因表达的影响,本研究以灭菌土壤为生长基质,将滇重楼(Paris polyphylla var. yunnanensis)新鲜种子与28种AM真菌于室温盆栽条件下共培养,采用实时荧光定量PCR的方法检测鲨烯环氧酶基因(Squalene epoxidase,SE)、共生受体类似激酶基因(Symbiosis-receptor-like kinase,SYMRK)、产生钙离子振荡的通道蛋白基因(Doesn’t making fections 1,DMI1)、钙/钙调依赖性蛋白激酶基因(Calcium/calmodulin-dependent protein kinase,CCa MK)4个功能基因在滇重楼幼苗的差异表达情况。结果表明:28株AM菌株可不同程度的影响SE、SYMRK、DMI1、CCa MK 4个功能基因的表达,其中薄壁两性囊霉(Ambisporaleptoticha,Ale)和崔氏原囊霉(Archaeospora trappei,Atr)可以显著增加4个功能基因的表达量。隐类球囊霉(Paraglo-mus occultum,Po)和透明盾巨孢囊霉(Scutellospora pellucida,Spe)可以增加SE、SYMRK的表达,细凹无梗囊霉(Acau-lospora scrobiculata,Asc),亮色盾巨孢囊霉(Racocetra fulgida,Rfu),哥伦比亚内养囊霉(Entrophospora colombiana,Ec),明球囊霉(Rhizophagus clarus,Rcl),根内球囊霉(Rhizophagus intraradices,Rin)可以增加DMI1的表达。综上所述,接种的28株AM真菌菌株中Ale和Atr能显著的提高4种功能基因的表达,结合前期与滇重楼无菌播种幼苗进行共生培养分析AM真菌对种子萌发及幼苗化学成分的影响,推测这两株菌可望能作为培育滇重楼菌根化苗的理想菌株,人工接种AM真菌可为保护和提高滇重楼开辟了一条新的途径。     

Yersinia pestis and plague: an updated view on evolution,virulence determinants,immune subversion,vaccination and diagnostics

Plague is a vector-borne disease caused by Yersinia pestis. Transmitted by fleas from rodent reservoirs, Y. pestis emerged less than 6000 years ago from an enteric bacterial ancestor through events of gene gain and genome reduction. It is a highly remarkable model for the understanding of pathogenic bacteria evolution, and a major concern for public health as highlighted by recent human outbreaks. A complex set of virulence determinants, including the Yersinia outer membrane proteins (Yops), the broad range protease Pla, pathogen-associated molecular patterns (PAMPs) and iron capture systems play critical roles in the molecular strategies that Y. pestis employs to subvert the human immune system, allowing unrestricted bacterial replication in lymph nodes (bubonic plague) and in lungs (pneumonic plague). Some of these immunogenic proteins as well as the capsular antigen F1 are exploited for diagnostic purposes, which are critical in the context of the rapid onset of death in the absence of antibiotic treatment (less than a week for bubonic plague and less than 48 h for pneumonic plague). In here, we review recent research advances on Y. pestis evolution, virulence factors function, bacterial strategies to subvert mammalian innate immune responses, vaccination and problems associated to pneumonic plague diagnosis.     

滇重楼寄生菌的研究

从滇重楼(Paris polyphylla var.yunnanensis)地下茎中分离和鉴定出两种细菌——蜡状芽孢杆菌(Bacillus cereus)和产碱假单胞菌(Pseudomonas alcaligenes),以及三种真菌——黑团孢霉(Periconia sp.)、白色厚顶孢霉(Pachnocybe albida)和重楼索霉(Hormomyces paridiphilus)。对蜡状芽孢杆菌、产碱假单胞菌和重楼索霉进行了液体培养并测定了胞外多糖含量,结果表明重楼索霉可分泌大量胞外多糖,这可能是导致滇重楼地下茎胶质化和多糖含量增加的原因。     

Detecting latitudinal and altitudinal expansion of invasive bamboo Phyllostachys edulis and Phyllostachys bambusoides (Poaceae) in Japan to project potential habitats under 1.5°C–4.0°C global warming

Rapid expansion of exotic bamboos has lowered species diversity in Japan's ecosystems by hampering native plant growth. The invasive potential of bamboo, facilitated by global warming, may also affect other countries with developing bamboo industries. We examined past (1975–1980) and recent (2012) distributions of major exotic bamboos (Phyllostachys edulis and P. bambusoides) in areas adjacent to 145 weather stations in central and northern Japan. Bamboo stands have been established at 17 sites along the latitudinal and altitudinal distributional limit during the last three decades. Ecological niche modeling indicated that temperature had a strong influence on bamboo distribution. Using mean annual temperature and sun radiation data, we reproduced bamboo distribution (accuracy = 0.93 and AUC (area under the receiver operating characteristic curve) = 0.92). These results infer that exotic bamboo distribution has shifted northward and upslope, in association with recent climate warming. Then, we simulated future climate data and projected the climate change impact on the potential habitat distribution of invasive bamboos under different temperature increases (i.e., 1.5°C, 2.0°C, 3.0°C, and 4.0°C) relative to the preindustrial period. Potential habitats in central and northern Japan were estimated to increase from 35% under the current climate (1980–2000) to 46%–48%, 51%–54%, 61%–67%, and 77%–83% under 1.5°C, 2.0°C, 3.0°C, and 4.0°C warming levels, respectively. These infer that the risk areas can increase by 1.3 times even under a 1.5°C scenario and expand by 2.3 times under a 4.0°C scenario. For sustainable ecosystem management, both mitigation and adaptation are necessary: bamboo planting must be carefully monitored in predicted potential habitats, which covers most of Japan.     

Qualitative and quantitative determination of major saponins in Paris and Trillium by HPLC-ELSD and HPLC–MS/MS

High-performance liquid chromatographic (HPLC) with evaporative light scattering detection (ELSD) and HPLC with electrospray ionization multistage tandem mass spectrometry (HPLC–ESI-MSⁿ) were used to identify and quantify steroid saponins in Paris and Trillium plants. The content of the known saponins such as Paris I, II, III, V, VI, VII, H, gracillin and protodioscin in Paris and Trillium plants was determined simultaneously using the developed HPLC-ELSD method. Furthermore, other 12 steroid saponins were identified by HPLC–ESI(+/−)-MSⁿ detection. In the end, a developed analytical procedure was proved to be a reliable and rapid method for the quality control of Paris and Trillium plants. In addition, the alternative resources for Paris yunnanensis used as a traditional Chinese medicine were discovered according to the hierarchical clustering analysis of the saponin fraction of these plants.     

Urinary cytology and the Paris system for reporting urinary cytology: Implications for urological management

By reducing the rate of indeterminate (atypical) diagnoses and standardising reporting terminology, The Paris System for Reporting Urine Cytology helps focus the application of cytology towards the detection primarily of high‐grade urothelial carcinoma. We present a urology‐based perspective of how the new system has influenced clinical decision‐making.     

Curcumin enhances the anti‐cancer effects of Paris Saponin II in lung cancer cells

Objectives

To investigate the synergistic mechanisms of Paris Saponin II (PSII) and Curcumin (CUR) in lung cancer.

Materials and Methods

The combination changed the cellular uptake of CUR and PSII, apoptosis, cell cycle arrest and cytokine levels were analysed on different lung cancer cells.

Results

The combination displayed a synergistic anti‐cancer effect through promoting the cellular uptake of CUR on different lung cancer cells. Hoechst H33258 staining and FACS assay indicated that the combination of PSII and CUR induced cell cycle arrest and apoptosis. Western blot and cytokine antibody microarray suggested that the combination activated death receptors such as DR6, CD40/CD40L, FasL and TNF‐α to induce cancer cells apoptosis, and up‐regulated IGFBP‐1 leading to inhibition of PI3K/Akt pathway and increase of p21 and p27, which therefore induced a G2 phase arrest in NCI‐H446 cells. Meanwhile, the combination suppressed PCNA and NF‐κB pathway in 4 kinds of lung cancer cells. They activated the phosphorylation of p38 and JNK, and inhibited PI3K in NCI‐H460 and NCI‐H446 cells, enhanced the phosphorylation of JNK in NCI‐H1299 cells, and increased the phosphorylation of p38 and ERK, and suppressed PI3K in NCI‐H520 cells.

Conclusions

PSII combined with CUR had a synergistic anti‐cancer effect on lung cancer cells. These findings provided a rationale for using the combination of curcumin and PSII in the treatment of lung cancer in future.