Biological mechanisms discriminating growth rate and adult body weight phenotypes in two Chinese indigenous chicken breeds.

BACKGROUND: Intensive selection has resulted in increased growth rates and muscularity in broiler chickens, in addition to adverse effects, including delayed organ development, sudden death syndrome, and altered metabolic rates. The biological mechanisms underlying selection responses remain largely unknown. Non-artificially-selected indigenous Chinese chicken breeds display a wide variety of phenotypes, including differential growth rate, body weight, and muscularity. The Wuding chicken breed is a fast growing large chicken breed, and the Daweishan mini chicken breed is a slow growing small chicken breed. Together they form an ideal model system to study the biological mechanisms underlying broiler chicken selection responses in a natural system. The objective of this study was to study the biological mechanisms underlying differential phenotypes between the two breeds in muscle and liver tissues, and relate these to the growth rate and body development phenotypes of the two breeds. RESULTS: The muscle tissue in the Wuding breed showed higher expression of muscle development genes than muscle tissue in the Daweishan chicken breed. This expression was accompanied by higher expression of acute inflammatory response genes in Wuding chicken than in Daweishan chicken. The muscle tissue of the Daweishan mini chicken breed showed higher expression of genes involved in several metabolic mechanisms including endoplasmic reticulum, protein and lipid metabolism, energy metabolism, as well as specific immune traits than in the Wuding chicken. The liver tissue showed fewer differences between the two breeds. Genes displaying higher expression in the Wuding breed than in the Daweishan breed were not associated with a specific gene network or biological mechanism. Genes highly expressed in the Daweishan mini chicken breed compared to the Wuding breed were enriched for protein metabolism, ABC receptors, signal transduction, and IL6-related mechanisms. CONCLUSIONS: We conclude that faster growth rates and larger body size are related to increased expression of genes involved in muscle development and immune response in muscle, while slower growth rates and smaller body size are related to increased general cellular metabolism. The liver of the Daweishan breed displayed increased expression of metabolic genes.

One-pot domino syntheses of 3-alkyl-3-N-substituted aminobenzofuran-2(3H)-ones based on alkali-promoted Michael addition and lactonization.

In this paper, a novel cascade reaction of caesium carbonate-promoted Michael addition and lactonization for the one-pot synthesis of 3-alkyl-3-N-substituted aminobenzofuran-2(3H)-one derivatives has been established based on the screening of the alkaline reagents and optimization of reaction conditions, in which the N-substituted (ortho-hydroxy)aryl glycine esters were used as the Michael donors to react with different α, ß-unsaturated carbonyl compounds. In the case of using the asymmetric starting material, the epimers could be successfully separated by conventional chromatography. In addition, plausible mechanisms were suggested and the absolute configuration of the epimer was analysed. All the chemical structures of unreported benzofuran-2(3H)-one derivatives were characterized by 1H nuclear magnetic resonance (NMR), 13C NMR, IR and high-resolution mass spectrometry (HRMS).

[Research on standardized preparation of traditional Chinese medicine (III): difference of extracting quantity of anthraquinones from mixed decoction of rhubarb with TCMs containing different ingredients].

OBJECTIVE: To detect the influence of compatibility of rhubarb with different traditional Chinese medicines (TCM) on extracted quantities of AQs, and to provide scientific basis for the clinical code for rhubarb preparation. METHOD: The influence of compatibility of rhubarb with different traditional Chinese medicines (saponin, alkaloids, flavonoids TCMs, animal medicines and mineral medicines) on decocting volume of anthraquinone substance was detected using ultra performance liquid chromatography. RESULT: In comparable conditions, more AQs were extracted from mixed decoction of rhubarb and saponin medicinal materials (Astragali Radix, Notoginseng Radix et Rhizoma, Glycyrrhizae Radix et Rhizoma, Polygalae Radix, Ginseng Radix et Rhizoma) than single decocting of rhubarb. The mixed decoction of rhubarb and alkaloid medicinal materials (Coptidis Rhizoma, Sophorae Flavescentis Radix, Prepared Aconiti Lateralis Radix Praeparata, Phellodendri Chinensis Cortex, Aconiti Lateralis Radix) caused a remarkable decrease in extracted quantities of AQs. And the mixed decoction of rhubarb and mineral medicines (Natrii Sulfas, Gypsum Fibrosum, Ostreae Concha, Alumen) also resulted in less extracted quantities of AQs to varying degrees. Besides, more rhubarb AQs were extracted from mixed decoction with Curcuma than single decoction. But less rhubarb AQs were observed in mixed decoction with Lonicerae Flos, Rehmanniae, Artemisiae Herb and Forsythiae Fructus than single decoction to varying degrees. In the study, the maximum extracted quantities of AQs is 2. 3-fold higher than the minimum, the largest difference existed in the extracted quantity of physcion which was 13.5 times. CONCLUSION: In compatibility between rhubarb and different TMCs, mixed decoction and single decoction show different influences on extracted quantity of rhubarb AQs. It is proved that more AQs may be extracted from mixed decoction between rhubarb and saponin medicinal materials, whereas less AQs may be observed in mixed decoction between rhubarb and alkaloid medicinal materials.

Improved U-Net based on contour prediction for efficient segmentation of rectal cancer.

BACKGROUND AND OBJECTIVE: Segmentation of rectal cancerous regions using 2D Magnetic Resonance Imaging (MRI) images is a critical step in radiation therapy. The shape of rectal cancer has significant variations and the shape of some surrounding organs is similar to that of rectal cancer; these conditions significantly affect the segmentation accuracy of rectal cancer and lead to incorrect segmentation. Therefore, automatic segmentation of rectal cancer is urgently needed, and it is a great challenge. For this task, the existing deep learning-based approaches have two shortcomings: 1) The U-Net network plays an important role in the field of medical segmentation. However, the designs of encoders and decoders in traditional U-Net networks are relatively simple and cannot extract good features, resulting in incorrect segmentation results. 2) Conventional neural networks extract high-level features that often do not include sufficient high-resolution contour information, resulting in ambiguity in contour segmentation. In this paper, we propose an improved U-Net network based on contour prediction, aiming at effective segmentation of rectal cancer. METHODS: We designed a new U-Net network by improving the traditional U-Net network. We made four improvements: 1) We replaced the encoders with the SENet network. 2) A global pooling layer was added after the last encoder. 3) We added the Spatial and Channel Squeeze & Excitation (SCSE) attention mechanism module to each decoder. 4) We concatenated the output results of each decoder. In addition, the model implemented content segmentation and contour segmentation for rectal cancer in parallel, so that both the content and contour information was learned by the network to enhance the segmentation accuracy. RESULTS: Our data were obtained from the Shanxi Provincial Cancer Hospital and included 3773 2D MRI rectal cancer images. The proposed method achieved an Mean Intersection over Union of 0.894 (MIoU) on the test set. Compared with state-of-the-art methods, our method had the best performance on the test set, and its MIoU metric was 0.123 higher than that of the second-best model. At the same time, the effectiveness of the improvements to our method was demonstrated through ablation experiments. CONCLUSIONS: Our method can help radiologists to segment effectively, save their time and energy, and enable them to focus on cases that are not easily segmented because of the complex shape of rectal cancer.

Aging affects artemisinin synthesis in Artemisia annua.

Artemisinin (ART) is the most effective component in malaria treatment, however, the extremely low content restricts its clinical application. Therefore, it is urgent to increase the yield of ART. ART gradually accumulates with aging, small RNA (sRNA) and transcriptome analysis were applied on the leaves of 2-week-old (2 w) and 3-month-old (3 m) A. annua respectively. Among all the annotated sRNAs, 125 were upregulated and 128 downregulated in the 3 m sample compared to the 2 w one. Whereas 2183 genes were upregulated and 2156 downregulated. Notably, the level of miR156 and several annotated miRNAs gradually decreased while SPLs increased. In addition, the genes on ART biosynthesis pathway were significantly upregulated including ADS, CYP71AV1, ADH1, DBR2 and ALDH1, and so were the positive transcription factors like AaERF1, AaORA and AaWRKY1 indicating that age influences the ART biosynthesis by activating the expression of the synthesizing genes as well as positive transcription factors. This study contributes to reveal the regulatory effects of age on ART biosynthesis both in sRNA and transcription levels.

[Antimicrobial activity and chemical differences between the two chemotypes of rhubarbs].

Through our pre-investigation and literature analysis, it was found that rhubarb could be categorized into two types, chrysophanol-type and rhein-type, based on the proportion of the two constituents in the total content of anthraquinones after acid hydrolysis. In this paper, the antimicrobial activities of chrysophanol-type and rhein-type rhubarbs against Staphylococcus aureus were compared with microcalorimetric analysis, in order to illustrate the bioactive differentiability between the two chemotypes. For the aim to display the distinction of chrysophanol and rhein percentage in total anthraquinones, the sampling volume was regulated to make the total anthraquinones equivalent, thus, the antimicrobial difference was only attributed to the difference of chemotypes. The results indicated that the antimicrobial difference between the two chemotypes was confirmable labeled at the biothermokinetic parameters of S. aureus growth affected by the rhubarb samples. The growth rate constant (k1) of the first exponential phase for the growth of S. aureus affected by the rhein-type rhubarb was significantly lower than that of chrysophanol-type (P<0.01), which suggested stronger antimicrobial activity of rhein-type rhubarb than that of chrysophanol-type. However, the antimicrobial activities of rhein-type rhubarbs were not positively correlated to the contents of rhein. It suggested that the antimicrobial activity of rhubarb might be related to some unknown components which were of same accumulating pattern of rhein. The findings in present study provided some experimental evidence on categorizing rhubarb into two chemotypes through the difference of antimicrobial activity on S. aureus by microcalorimetric analysis and, further, offered references to revision of the commercial specification of rhubarb from chemical view.

[Rationality of commercial specification of rhubarb based on chemical analysis].

The differences of 34 rhubarb samples collected on the market and at producing area were investigated by chemical analysis on the contents of anthraquinones and chromatographic fingerprints, in order to assess the rationality of the commercial specification of rhubarb. The results indicated that the commercial specification of rhubarb was not correlated to the contents of anthraquinones as well as the price. The chromatographic fingerprints of rhubarb samples from different producing area were dissimilar, while the commercial specifications were difficult to be separated. Generally, the rhubarb samples produced in famous-region contained more anthraquinones. This demonstrated rationality on the traditional records of the famous-region of rhubarb from a chemical view. In this study, it was firstly reported that rhubarb could be categorized into two types, chrysophanol-type and rhein-type, based on the proportion of the two constituents in the total anthraquinones after acid hydrohysis. It was found that the rhubarb samples of rhein-type were mostly produced in famous-regions, such as Qinghai, Xizang, West Sichuan and Gansu. The literatures reported that rhein was superior to chrysophanol at many pharmacological effects and pharmacokinetic properties. Hence, we primarily considered that rhein-type rhubarb might be high-quality. These results were helpful to improve the commercial specification of rhubarb from a view of chemical information.

[Delphi-based study on scientificity of organoleptic evaluation for rhubarb commercial specification].

OBJECTIVE: In this study the reliability and scientificity of organoleptic evaluation for the commercial specification of Chinese medicinal materials (CMM) was investigated with Delphi method, using rhubarb as an example. The consensus panel of panelists being expert in organoleptic evaluation for CMM was cogitatively selected, with high average authority index of 0.81. The average individual repeatability of selected panelists was 85%, while the group repeatability was above 70%. It was found that the career time of the expert had significant influence on the evaluation result (P < 0.05), which suggested that practice makes repeatability. Some panelists of different career type and education background concluded different results, but no statistical significance was found. Furthermore, four predominant organoleptic parameters, shape, color, texture and section characteristics, were selected based on statistics and panelists' discussion to judge the commercial specification of rhubarb. In conclusion, we firstly demonstrated the organoleptic evaluation is an objective and repeatable approach to judge the commercial specification of CMM; and we recommend Delphi method to do validation and standardization for the conventional experiences of traditional Chinese medicine.

Enhanced anticoagulant activity of hirudin-i analogue co-expressed with arylsulfotransferase in periplasm of E. coli BL21(DE3).

Hirudin, a blood anticoagulant, is the most potent natural thrombin inhibitor of leech origin. Its application is limited because it is difficult to obtain abundant natural hirudin directly from the leech. Although some bioengineering methods can significantly increase the production of hirudin, the reduced efficacy of recombinant hirudin (rH) remains a critical shortcoming. The lack of sulfation of tyrosine 63 in rH is an important cause of its inadequate performance. This article is the first report of periplasmic co-expression of an rH-I analogue with arylsulfotransferase (ASST) in E. coli BL21(DE3). Co-expressed rH-I analogue with sulfate donor substrate (p-nitrophenyl sulfate potassium) showed anticoagulant (rabbit and goat serum) activity twice more than rH-I analogue expressed without ASST, indicating its potential periplasmic sulfation. Moreover, purified rH-I analogue showed above 4.5 times higher anticoagulant activity compared to therapeutic anti-thrombotic heparin (HE). At the same time, pH-dependent differential solubility was employed to purify rH analogues from fermentation broth, which is a simple, fast and inexpensive purification technology, and can potentially be used for larger scale purification. This will also greatly improve the application of rH in clinical treatment.

Comparison of hybridization behavior between double and single strands of targets and the application of asymmetric PCR targets in cDNA microarray.

Double stranded targets on the cDNA microarray contain representatives of both the coding and noncoding strands, which will introduce hybridization competition with probes. Here, the effect of double and single strands of targets on the signal intensity and the ratios of Cy5/Cy3 within the same slide were compared. The results show that single stranded targets can increase the hybridization efficiency without changing the Cy5/Cy3 ratio. Based on these results, a new strategy was established by generating cDNA targets with asymmetric PCR, instead of conventional PCR, to increase the sensitivity of the cDNA microarray. Furthermore, the feasibility of this approach was validated. The results indicate that the cDNA microarray system based on asymmetric PCR is more sensitive, with no decrease in the reliability and reproducibility as compared with that based on conventional symmetric PCR.

Co-expression of Erns and E2 genes of classical swine fever virus by replication-defective recombinant adenovirus completely protects pigs against virulent challenge with classical swine fever virus.

The objective of this study was to construct a recombinant adenovirus for future CSFV vaccines used in the pig industry for the reduction of losses involved in CSF outbreaks. The Erns and E2 genes of classical swine fever virus (CSFV), which encode the two main protective glycoproteins from the "Shimen" strain of CSFV, were combined and inserted into the replication-defective human adenovirus type-5 and named the rAd-Erns-E2. Nine pigs were randomly assigned to three treatment groups (three pigs in each group) including the rAd-Erns-E2, hAd-CMV control and DMEM control. Intramuscular vaccination with 2×10(6) TCID(50) of the rAd-Erns-E2 was administered two times with an interval of 21 days. At 42 days post inoculation, pigs in all groups were challenged with a lethal dose of 1×10(3) TCID(50) CSFV "Shimen" strain. Observation of clinical signs was made and the existence of CSFV RNA was detected. Animals in the hAd-CMV and DMEM groups showed severe clinical CSF symptoms and were euthanized from 7 to 10 days after the challenge. However, no adverse clinical CSF signs were observed in vaccinated pigs after the administration of rAd-Erns-E2 and even after CSFV challenge. Neither CSFV RNA nor pathological changes were detected in the tissues of interest of the above vaccinated pigs. These results implied that the recombination adenovirus carrying the Erns-E2 genes could be used to prevent swine from classical swine fever.

Colorimetric grading scale can promote the standardization of experiential and sensory evaluation in quality control of traditional Chinese medicines.

Experiential and sensory evaluation is an ancient method that remains important in the current quality control system of Traditional Chinese Medicines (TCMs). The process is rapid and convenient when evaluating the quality of crude materials in TCM markets. However, sensory evaluation has been met with skepticism because it is mainly based on experience and lacks a scientific basis. In this study, rhubarb was selected to demonstrate how color-based sensory evaluation could differentiate the quality of herbal medicines objectively. The colors of the rhubarb samples, expressed as RGB values, were obtained from different parts and forms of the plant, including the plant's surface, fracture surface color, and a powdered form with or without treatment with a color-developing reagent. We first divided the rhubarb samples into three grades based on the total content of five hydroxyanthraquinone derivatives, the major pharmacological components in rhubarb. Then, a three-layer back-propagation artificial neural network (BP-ANN), calibrated with selected training samples, was used to correlate the quality of the rhubarb with its color. The color of the rhubarb powder after coloration attained the highest accuracy (92.3%) in predicting the quality grade of the test samples with the established artificial neural networks. Finally, a standardized colorimetric grading scale was created based on the spatial distribution of the rhubarb samples in a two-dimensional chromaticity diagram according to the colors of the powdered rhubarb after color enhancement. By comparing the color between the scale and the tested samples, similar to performing a pH test with indicator paper, subjects without sensory evaluation experience could quickly determine the quality grade of rhubarb. This work illustrates the technical feasibility of the color-based grading of rhubarb quality and offers references for quantifying and standardizing the sensory evaluation of TCMs, foods and other products.