Effect of Bushen Qudu Decoction on TGF-β1/Smads signal transduction pathway in rats with chronic renal failure

Purpose: To study the impact of Bushen Qudu Decoction on renal function, renal tissue morphology and TGF-β1/Smads signal transduction pathway in adenine-induced chronic renal failure rats.Methods: 76 rats were assigned to normal group, model group, uremic clearance group, benazepil group, Bushen Qudu decoction gunshan A group and Bushen Qudu decoction Guiza group. Renal failure was induced in rats using intragastric administration of adenine for 30 days. The expressions of TGF-β1, Smad2, Smad3 and Smad6 were determined with immunohistochemistry. Real-time quantitative PCR was employed to assay the mRNA expression of TGF-β1 in each group, while protein expressions of Smad2, Smad3 and Smad6 were determined with western blot. The scores of glomerular and tubulointerstitial lesions in each group were obtained by histopathological examination.Results: Bushen Qudu decoction significantly reduced BUN and creatinine in rats with chronic renal failure; furthermore, it also lowered the protein expressions of TGF-β1 and SMAD2/3, but Smad6 protein, relative to control (p < 0.05). The TGF-β1mRNA expression was down-regulated (p < 0.05), relative to control group.Conclusion: Bushen Qudu decoction reduces renal interstitial fibrosis in rats and delays the progression of chronic renal failure by regulating TGF-β1/Smads signaling pathway. These findings provide some insight that should facilitate the development of new drugs that would delay the onset of chronic renal failure

Rapid Determination of Saponins in the Honey-Fried Processing of Rhizoma Cimicifugae by Near Infrared Diffuse Reflectance Spectroscopy.

ObjectiveA model of Near Infrared Diffuse Reflectance Spectroscopy (NIR-DRS) was established for the first time to determine the content of Shengmaxinside I in the honey-fried processing of Rhizoma Cimicifugae.MethodsShengmaxinside I content was determined by high-performance liquid chromatography (HPLC), and the data of the honey-fried processing of Rhizoma Cimicifugae samples from different batches of different origins by NIR-DRS were collected by TQ Analyst 8.0. Partial Least Squares (PLS) analysis was used to establish a near-infrared quantitative model.ResultsThe determination coefficient R² was 0.9878. The Cross-Validation Root Mean Square Error (RMSECV) was 0.0193%, validating the model with a validation set. The Root Mean Square Error of Prediction (RMSEP) was 0.1064%. The ratio of the standard deviation for the validation samples to the standard error of prediction (RPD) was 5.5130.ConclusionThis method is convenient and efficient, and the experimentally established model has good prediction ability, and can be used for the rapid determination of Shengmaxinside I content in the honey-fried processing of Rhizoma Cimicifugae

Simple and efficient expression of codon-optimized mouse leukemia inhibitory factor in Escherichia coli

Purpose: To obtain a higher yield of mouse leukemia inhibitory factor to maintain the proliferation potential of pluripotent stem cells at a low cost.Methods: A method was designed to produce recombinant mLIF protein (rmLIF) in Escherichia coli. Through analysis of rmLIF sequence, it was found that rare codons were interspersed. After mutation from rare codons to Escherichia coli (E. coli) preferred ones were selected, the mutated gene mLIFm was cloned into pET15b vector. The pET15b-mLIFm was then transformed into Rosetta-gami strain and induced with optimal conditions at 18 oC for 16 h. Mass spectrometry was carried out to identify the peptides.Results: After purification, the yield of the codon-optimized rmLIFm was 141 mg/L, compared with 110 mg/L for the original rmLIF. Mass spectral analysis showed the presence of four major peptides each with an intensity > 10 % at m/z 1031.57, 1539.82, 1412.01 and 2229.10 in mLIFm, respectively. Histagged rmLIFm fusion protein displayed the potential to maintain the morphology of undifferentiated mouse embryonic stem cells (mESCs), which were positive for mESCs markers (Oct-4, Nanog, Sox-2, stage-specific embryonic antigen-1).Conclusion: The findings provide a means to produce mLIF in a short, useful, cost-effective and environmentally-friendly manner, and thus lays a foundation for further studies of mLIF.Keywords: Leukemia inhibitory factor, Mutated gene, Protein expression, Purification, Stem cells, Peptides, Escherichia col

Inhibitory Effects of Columbianadin on Nociceptive Behaviors in a Neuropathic Pain Model, and on Voltage-Gated Calcium Currents in Dorsal Root Ganglion Neurons in Mice

Radix angelicae pubescentis (RAP) has been used in Chinese traditional medicine to treat painful diseases such as rheumatism and headache. A previous study has reported that columbianadin (CBN), a major coumarin in RAP inhibits acute and inflammatory pain behaviors. However, the effects of CBN on neuropathic pain behaviors, and the potential underlying mechanism have not been reported. In the present study, the effects of CBN, compared to another major coumarin of RAP osthole (OST), on oxaliplatin-induced neuropathic pain behaviors and on the voltage-gated calcium currents in small dorsal root ganglion (DRG) neurons were studied in mice. It was found that CBN and OST inhibited both mechanical and cold hypersensitivity induced by oxaliplatin. Moreover, CBN and OST might preferentially inhibit T- and L-type calcium currents (Ica). The inhibitory effects of CBN and OST on the oxaliplatin-induced mechanical allodynia were prevented by gabapentin. These results suggest that CBN, as well as OST might inhibit neuropathic pain behaviors through an inhibition of T- and L-type calcium currents in nociceptive DRG neurons

Genetic Differentiation and Delimitation between Ecologically Diverged Populus euphratica and P. pruinosa

The fixed genetic differences between ecologically divergent species were found to change greatly depending on the markers examined. With such species it is difficult to differentiate between shared ancestral polymorphisms and past introgressions between the diverging species. In order to disentangle these possibilities and provide a further case for DNA barcoding of plants, we examine genetic differentiation between two ecologically divergent poplar species, Populus euphratica Oliver and P. pruinosa Schrenk using three different types of genetic marker.We genotyped 290 individuals from 29 allopatric and sympatric populations, using chloroplast (cp) DNA, nuclear (nr) ITS sequences and eight simple sequence repeat (SSR) loci. Three major cpDNA haplotypes were widely shared between the two species and between-species cpDNA differentiation (F(CT)) was very low, even lower than among single species populations. The average SSR F(CT) values were higher. Bayesian clustering analysis of all loci allowed a clear delineation of the two species. Gene flow, determined by examining all SSR loci, was obvious but only slightly asymmetrical. However, the two species were almost fixed for two different nrITS genotypes that had the highest F(CT), although a few introgressed individuals were detected both in allopatric and sympatric populations.The two species shared numerous ancestral polymorphisms at cpDNA and a few SSR loci. Both ITS and a combination of nuclear SSR data could be used to differentiate between the two species. Introgressions and gene flow were obvious between the two species either during or after their divergence. Our findings underscore the complex genetic differentiations between ecologically diverged species and highlight the importance of nuclear DNA (especially ITS) differentiation for delimiting closely related plant species

Scientometric review of construction project schedule studies: trends, gaps and potential research areas

Scheduling plays a fundamental role in construction projects’ success and thus has drawn attention from both academic researchers and industry practitioners. A large number of research articles tend to solve emerging challenges in construction project schedule (CPS). Therefore, there is a strong need of systematic review on existing studies. In this study, a total of 332 articles were retrieved from Scopus database using title, abstract and keywords with respect to CPS and filtered by document type, language type and abstract content. In particular, science mapping approach was adopted to analyse selected journal articles. These articles were examined using three sequential processes, including bibliometric search, scientometric analysis, and in-depth qualitative discussion. It could demonstrate the most influential journals, researchers, published articles, and active countries/regions in this area. In addition, major CPS knowledge areas were identified and summarized as CPS constructability, applications of variety of CPS methods, CPS optimization models and algorithms, identification and quantification of schedule risks and uncertainties, CPS performance management, and adopting new emerging CPS technologies and methods. Furthermore, knowledge gaps and future potential research directions were also discussed in detail. Finally, a comprehensive CPS framework was proposed as a sound reference in future research

Analysis of labor resources wastage in China’s real estate brokerage: from the perspective of opportunity costs

Real estate brokerage has experienced the rapid growth over the past two decades in China, with a significant increase of employees. In particular, in the megacities like Beijing, the growth of employees exceeds the growth of real estate transaction volume. This may lead to the wastage of labor resources. In this regard, the optimal employee size (OES) in China’s real estate brokerage is proposed from the perspective of opportunity costs, which include both under-size and over-size costs. In the proposed OES models, a real estate brokerage firm makes the optimal decisions of number of employees by minimizing expected opportunity costs. In addition, an iterative algorithm is employed to obtain the optimal employee size in different scenarios. The result reveals that high profit gained from the business does attract more employees than what is needed. By addressing various scenarios based on the game model, it is found that asymmetric competition, the increase of market participants, and demand fluctuations also contribute to the labor resources wastage in real estate brokerage industry. The theoretical analysis results are verified by taking Beijing as the case study. Finally, suggestions for reducing labor resources wastage in real estate brokerage of China are provided

A Novel α-Galactosidase A Splicing Mutation Predisposes to Fabry Disease

Fabry disease (FD) is a rare X-linked α-galactosidase A (GLA) deficiency, resulting in progressive lysosomal accumulation of globotriaosylceramide (Gb3) in a variety of cell types. Here, we report a novel splicing mutation (c.801 + 1G > A) that results in alternative splicing in GLA of a FD patient with variable phenotypic presentations of renal involvement. Sequencing of the RT-PCR products from the patient’s blood sample reveals a 36-nucleotide (nt) insertion exists at the junction between exons 5 and 6 of the GLA cDNA. Splicing assay indicates that the mutated minigene produces an alternatively spliced transcript which causes a frameshift resulting in an early termination of protein expression. Immunofluorescence shows puncta in cytoplasm for mutated GLA whereas uniform staining small dots evenly distributed inside cytoplasm for wild type GLA in transfected HeLa cells. The increased senescence and decreased GLA enzyme activity suggest that the abnormalities might be due to the altered localization which further might result from the lack of the C-terminal end of GLA. Our study reveals the pathogenesis of splicing mutation c.801 + 1G > A to FD and provides scientific foundation for accurate diagnosis and precise medical intervention for FD