Effects of Different Cultivation Modes on Morphological Traits and Correlations between Traits and Body Mass of Crayfish (Procambarus clarkii).

In this study, juvenile crayfish hatched from the same population were cultured in different growing environments: pond (D1), paddy field (D2), and aquaculture barrel (D3), and fed for 60 days. Crayfishes were selected randomly, females and males, 50 tails each from six groups (D1-♀, D1-♂, D2-♀, D2-♂, D3-♀, D3-♂) to measure the following morphological traits: full length (X1), body length (X2), chelicerae length (X3), chelicerae weight (X4), cephalothorax length (X5), cephalothorax width (X6), cephalothorax height (X7), eye spacing (X8), caudal peduncle length (X9), and caudal peduncle weight (X10). We found that the coefficient of variation (CV) of X4 was the largest in each culture mode, and males (28.58%~38.67%) were larger than females (37.76%~66.74%). The CV of X4 of crayfish cultured in D1 and D2 was larger than that of D3. All traits except X8 were positively correlated with body weight (p < 0.05). After pathway analysis, we found that X4, X5, X7, and X10 were significantly correlated with the body weight of D1-♀; the equation was YD1-♀ = -29.803 + 1.249X4 + 0.505X5 + 0.701X7 + 1.483X10 (R2 = 0.947). However, X2, X4, and X6 were significantly correlated with the body weight of D1-♂; the equation was YD1-♂ = -40.881 + 0.39X2 + 0.845X4 + 1.142X6 (R2 = 0.927). In D2-♀, X1, X4, X5, and X10 were significantly correlated with body weight; the equation was YD2-♀ = -12.248 + 0.088X1 + 1.098X4 + 0.275X5 + 0.904X10 (R2 = 0.977). X4 and X5 played a major role in the body weight of D2-♂ with the equation: YD2-♂ = -24.871 + 1.177X4 + 0.902X5 (R2 = 0.973). X3 and X10 mainly contributed to the body weight of D3-♀ with the equation: YD3-♀ = -22.476 + 0.432X3 + 3.153X10 (R2 = 0.976). X1 and X4 mainly contributed to the body weight of D3-♂ with the equation: YD3-♂ = -34.434 + 0.363X1 + 0.669X4 (R2 = 0.918). Comparing the pathway analysis with the gray relation analysis, we could conclude that the traits most correlated with body weight in D1-♀ were X10 and X7; in D1-♂, X6; in D2-♀, X10, X1, and X5; in D2-♂, X5; in D3-♀, X10; and in D3-♂, X4 and X1.

Systematic screening of hepatoprotective components from traditional Chinese medicine: Zuojin Pill as an example.

ETHNOPHARMACOLOGICAL RELEVANCE: Zuojin Pill (ZJP), composed of Coptis chinensis Franch. and Euodia ruticarpa (A. Juss.) Benth. in a mass ratio of 6:1, is a famous traditional Chinese medicine (TCM) formula recorded in "Danxi's Experiential Therapy", an ancient medical book from the Ming Dynasty of China. It is used to treat liver fire invading the stomach, which is caused by liver stagnation transforming into fire and disharmony between the liver and stomach. AIM OF THE STUDY: To develop a systematic strategy to screen hepatoprotective components from TCM using ZJP as a model sample. MATERIALS AND METHODS: A CCl4-induced mouse model of acute liver injury was used for the verification of the hepatoprotective effects of ZJP. UPLC-Q-Exactive Plus Orbitrap MS/MS was used for the identification of the components in mouse serum after intragastric administration of ZJP. The hepatoprotective activities of the components found in mouse serum were tested in primary cultured mouse hepatocytes induced by CCl4. RESULTS: Nine components with significant hepatoprotective activity including berberine, epiberberine, coptisine, palmatine, jatrorrhizine, rutaecarpin, dehydroevodiamine, evocarpine and chlorogenic acid were successfully screened out. CONCLUSIONS: Our developed strategy has the advantages of high efficiency and low cost, and would provide a powerful tool for screening potential hepatoprotective components from TCM.

Study on quality control of Zuojin pill by HPLC fingerprint with quantitative analysis of multi-components by single marker method and antioxidant activity analysis.

Current quality control methods for Zuojin Pill (ZJP) lack comprehensiveness and practicability. This study aimed to develop a comprehensive strategy for the quality evaluation of ZJP and the prediction of potential bioactive components in ZJP. First, an HPLC method with excellent separation of main components was developed and was used to establish the chromatographic fingerprint of ZJP. Similarities were calculated by comparing 28 batches of ZJPs with the reference fingerprint and the resulting similarity values were all greater than 0.976. The 28 samples were classified into different groups according to their origins by Hierarchical Cluster Analysis, Principal component analysis, and orthogonal partial least squares discriminant analysis. Based on the classification, eight quality markers (Q-Markers) affecting the quality of ZJP were discovered. Then, using berberine as an internal standard substance, quantitative analysis of multi-components by single marker method (QAMS) for the determination of eight Q-markers was developed. The results showed that there was no significant difference between QAMS and external standard method (P＞0.05). Finally, using an off-line antioxidant system and partial least-squares model (PLS), the fingerprint-efficacy relationship of ZJP was constructed to explore and predict the bioactive components in ZJP. The present study strategy could be also applied to comprehensive quality study of other TCMs.

National-scale urinary phthalate metabolites in the general urban residents involving 26 provincial capital cities in China and the influencing factors as well as non-carcinogenic risks.

Phthalates (PAEs) are widely used in daily products but can cause a variety of adverse effects in humans. Few studies have been carried out on human internal exposure levels of PAEs on a large-scale, especially in developing countries. In the present study, 1161 urine samples collected from residents of 26 provincial capitals in China were analyzed for nine phthalate metabolites (mPAEs). The chemicals were widely detected, and the median specific gravity adjusted urinary concentration of Σ9mPAEs was 278 µg/L. Di-(2-ethylhexyl) phthalate (DEHP) and di-n-butyl phthalate (DnBP) were the main parent PAEs that the residents were exposed to. Demographic characteristics, such as age and educational level, were significantly associated with PAE exposure. Children and the elderly had higher mPAE levels. Subjects with lower educational levels were more frequently exposed to DnBP and DEHP. However, mono-ethyl phthalate showed the opposite trend, i.e., higher concentrations in subjects aged 18-59 years and with higher educational levels. Geographic differences were detected at the national scale. Residents in northeastern and western China had higher levels of mPAEs than those in central China, most likely because of different industrial usage of the chemicals and different living habits and living conditions of the residents. Health risk assessment showed that hazard indices of PAEs ranged from 0.07 to 9.34, with 20.0% of the subjects being concern for potential non-carcinogenic risk as assessed by Monte Carlo simulation. DEHP and DnBP were the primary contributors, representing 96.7% of total risk. This first large-scale study on PAE human internal exposure in China provides useful information on residents' health in a developing country, which could be used for chemical management and health protection.

Fast Screening of Biomembrane-Permeable Compounds in Herbal Medicines Using Bubble-Generating Magnetic Liposomes Coupled with LC-MS.

A novel strategy based on the use of bionic membrane camouflaged magnetic particles and LC-MS was developed to quickly screen the biomembrane-permeable compounds in herbal medicines. The bionic membrane was constructed by bubble-generating magnetic liposomes loaded with NH4HCO3 (BMLs). The lipid bilayer structure of the liposomes enabled BMLs to capture biomembrane-permeable compounds from a herbal extract. The BMLs carrying the compounds were then separated from the extract by a magnetic field. Upon heat treatment, NH4HCO3 rapidly decomposed to form CO2 bubbles within the liposomal bilayer, and the captured compounds were released from BMLs and analyzed by LC-MS. Jinlingzi San (JLZS), which contains various natural ingredients, was chosen to assess the feasibility of the proposed method. As a result, nine potential permeable compounds captured by BMLs were identified for the first time. Moreover, an in vivo animal study found that most of the compounds screened out by the proposed method were absorbed into the blood. The study provides a powerful tool for rapid and simultaneous prediction of multiple biomembrane-permeable components.

Conducting large, repeated, multi-game economic experiments using mobile platforms.

We demonstrate the possibility of conducting synchronous, repeated, multi-game economic decision-making experiments with hundreds of subjects in-person or remotely with live streaming using entirely mobile platforms. Our experiment provides important proof-of-concept that such experiments are not only possible, but yield recognizable results as well as new insights, blurring the line between laboratory and field experiments. Specifically, our findings from 8 different experimental economics games and tasks replicate existing results from traditional laboratory experiments despite the fact that subjects play those games/task in a specific order and regardless of whether the experiment was conducted in person or remotely. We further leverage our large subject population to study the effect of large (N = 100) versus small (N = 10) group sizes on behavior in three of the scalable games that we study. While our results are largely consistent with existing findings for small groups, increases in group size are shown to matter for the robustness of those findings.

Improving rare disease classification using imperfect knowledge graph.

BACKGROUND: Accurately recognizing rare diseases based on symptom description is an important task in patient triage, early risk stratification, and target therapies. However, due to the very nature of rare diseases, the lack of historical data poses a great challenge to machine learning-based approaches. On the other hand, medical knowledge in automatically constructed knowledge graphs (KGs) has the potential to compensate the lack of labeled training examples. This work aims to develop a rare disease classification algorithm that makes effective use of a knowledge graph, even when the graph is imperfect. METHOD: We develop a text classification algorithm that represents a document as a combination of a "bag of words" and a "bag of knowledge terms," where a "knowledge term" is a term shared between the document and the subgraph of KG relevant to the disease classification task. We use two Chinese disease diagnosis corpora to evaluate the algorithm. The first one, HaoDaiFu, contains 51,374 chief complaints categorized into 805 diseases. The second data set, ChinaRe, contains 86,663 patient descriptions categorized into 44 disease categories. RESULTS: On the two evaluation data sets, the proposed algorithm delivers robust performance and outperforms a wide range of baselines, including resampling, deep learning, and feature selection approaches. Both classification-based metric (macro-averaged F1 score) and ranking-based metric (mean reciprocal rank) are used in evaluation. CONCLUSION: Medical knowledge in large-scale knowledge graphs can be effectively leveraged to improve rare diseases classification models, even when the knowledge graph is incomplete.

Base composition of the Danio chrysotaenitus complete mitochondrial genome.

In this study, 16 sets of primers were used to amplify contiguous, overlapping segments of the complete mitochondrial DNA (mtDNA) of Danio chrysotaenitus in order to characterize and compare their mitochondrial genomes. The total length of the mitochondrial genome is 16,608 bp and deposited in the GenBank with accession numbers KP407138. The organization of the mitochondrial genomes was similar to those reported from other Mountain carp fishes mitochondrial genomes containing 37 genes (13 protein-coding genes, 2 ribosomal RNA, and 22 transfer RNAs) and a major non-coding control region (D-loop region). Most genes were encoded on the H-strand, except for the ND6 and 8 tRNA genes, encoding on the L-strand. The nucleotide skewness for the coding strands of Danio chrysotaenitus (AT-skew = 0.10, GC-skew = -0.25) is biased toward T and G. The complete mitogenome may provide important date set for the study of genetic mechanism of Danio chrysotaenitus.

The complete mitochondrial genome of the hybrid Cyprinus capio Furong. [Cyprinus carpio L. mirror (♀)×Cyprinus Carpio var. singuonensis (♂)].

The complete mitochondrial genome of the hybrid of Cyprinus carpio L. mirror (♀)× Cyprinus Carpio var. singuonensis (♂) was characterized first in this study. The total length of the genome was identical to the female parent as 16,581 bp, and the overall base composition was 31.80% A, 24.85% T, 27.55% C and 15.80% G, with a slight A + T bias. It contained 13 protein-coding genes (PCGs), 22 transfer RNA genes, 2 ribosomal RNA genes and 2 main non-coding regions (the control region and the origin of the light-strand replication). This study discovered the 99.70% sequence identity between the hybrid and its female parent, which confirmed the maternal inheritance pattern followed by the mitochondrial genome of the hybrid. However, the sequence alignment of mitochondrial genomes between the hybrid and its female parent revealed a total of 47 variable sites in 15 genes or regions, especially 25 sense mutations in 6 PCGs. The complete mitochondrial genome sequence of the hybrid Cyprinus capio Furong may provide an important dataset for further study in mitochondrial inheritance mechanism.

The complete mitochondrial genome of the hybrid Furong-crucian [Cyprinus capio Furong (♀) × Carassius auratus red var (♂)].

The complete mitochondrial genome of the hybrid of Cyprinus capio Furong (♀) × Carassius auratus red var. (♂) was characterized first in this study. The total length of the mitochondrial genome of Furong-crucian was identical to the female parent as 16,581 bp, and the overall base composition was 31.87% A, 24.81% T, 27.56% C and 15.76% G, with a slight A + T bias. It contained 13 protein-coding genes (PCGs), 22 transfer RNA genes, 2 ribosomal RNA genes and 2 main non-coding regions (the control region and the origin of the light-strand replication). This study discovered the 99.3% sequence identity between the hybrid and its female parent, which confirmed the maternal inheritance pattern followed by the mitochondrial genome of the hybrid. However, the sequence alignment of mitochondrial genomes between the hybrid and its female parent revealed a total of 109 variable sites in 18 genes or regions, especially 32 sense mutations in 9 PCGs. The complete mitochondrial genome sequence of this hybrid Furong-crucian may provide an important dataset for further study in mitochondrial inheritance mechanism.

Thyroxine and reserpine-induced changes in metabolic profiles of rat urine and the therapeutic effect of Liu Wei Di Huang Wan detected by UPLC-HDMS.

The promise of metabonomics, a new "omics" technique, to validate Chinese medicines and the compatibility of Chinese formulas has been appreciated. The present study was undertaken to explore the excretion pattern of low molecular mass metabolites in the male Wistar-derived rat model of kidney yin deficiency induced with thyroxine and reserpine as well as the therapeutic effect of Liu Wei Di Huang Wan (LW) and its separated prescriptions, a classic traditional Chinese medicine formula for treating kidney yin deficiency in China. The study utilized ultra-performance liquid chromatography/electrospray ionization synapt high definition mass spectrometry (UPLC/ESI-SYNAPT-HDMS) in both negative and positive electrospray ionization (ESI). At the same time, blood biochemistry was examined to identify specific changes in the kidney yin deficiency. Distinct changes in the pattern of metabolites, as a result of daily administration of thyroxine and reserpine, were observed by UPLC-HDMS combined with a principal component analysis (PCA). The changes in metabolic profiling were restored to their baseline values after treatment with LW according to the PCA score plots. Altogether, the current metabonomic approach based on UPLC-HDMS and orthogonal projection to latent structures discriminate analysis (OPLS-DA) indicated 20 ions (14 in the negative mode, 8 in the positive mode, and 2 in both) as "differentiating metabolites".

In vitro study of the effects of lens extract on rat retinal neuron survival and neurite outgrowth.

BACKGROUND: Optic nerve regeneration has previously been achieved by injuring the lens, which results in the release of lentogenic factors. However, these lentogenic factors are still unknown. OBJECTIVES: To investigate what were the lentogenic factors by examining the effects of lens extract and macrophage-conditioned medium (MCM) on the survival and the neurite outgrowth of rat retinal neurons in vitro. METHODS: Retinal neurons were cultured in 4 groups: (1) Dulbecco's modified Eagle's medium (DMEM), (2) DMEM containing lens extract, (3) DMEM containing macrophage-conditioned medium (MCM-D), (4) DMEM and medium from macrophages grown with lens extract (MCM-L). Neurite outgrowth and neuron survival time were observed. The density of retinal neurons with neurites and the longest neurites of the cells were measured on days 1, 3 and 5. RESULTS: Retinal neurons survive for 12-14 days in DMEM containing lens extract. However, the cells only survive for 6 days in DMEM and only 7 days in DMEM containing MCM-L or MCM-D. The present results indicate that lens extract may directly promote survival of rat retinal neurons and neurite outgrowth in vitro. The MCM also promoted cell survival and neurite outgrowth but its effects were weaker than that of the lens extract. We postulate that lens extract exerts its effect by direct neurotrophic effects and/or indirectly by activating macrophages in vitro.

Distribution and expression of RhoA in rat retina after optic nerve injury.

BACKGROUND: RhoA is a small guanosine triphosphatase which participates in signaling pathways of axonal repellents or inhibitors. However, the distribution and expression of RhoA in the rat retina after optic nerve injury has not been elucidated yet. OBJECTIVES: To study the distribution and expression of RhoA in the rat retina after optic nerve injury. METHODS: Immunohistochemistry was used to determine the distribution of RhoA in rat retina after optic nerve injury. The expression of RhoA was analyzed by Western blot. RESULTS: In normal retina and the retina 1 day after optic nerve injury, RhoA was distributed in the retinal ganglion cell (RGC) layer. Three days after optic nerve injury, it existed in RGCs and the inner plexiform layer. However, 7 days after surgery its immunoreactivity was abundant not only in the RGC and inner plexiform layers but also in the inner nuclear and outer plexiform layers. Western blot analysis showed that the expression of RhoA increased significantly in the retina after optic nerve injury in comparison with normal retina. CONCLUSION: These results indicate that the distribution and expression of RhoA were extended and enhanced after optic nerve injury, and that RhoA plays an important role in optic nerve regeneration.

[Influence of fluorescent protein expression on the proliferation of NIH3T3 cells in vitro].

OBJECTIVE: To investigate the influence of fluorescent protein expression on the proliferation of murine NIH3T3 cells, so as to provide a theoretical basis for cell tracing technology. METHODS: NIH3T3 cells were cultured in vitro, and were randomly divided into control, pLEGFP-N1 (with transfection of pLEGFP-N1 retroviral vector), pEGFP-N1 (with transfection of pEGFP-N1 vector) and pDsRed2-C1 (with transfection of pDsRed2-C1 vector) groups. Then the cells were screened by G418 for 3 weeks. The changes in cell adhesive rate were observed and the population doubling times was determined by growth curve. RESULTS: There was obvious fluorescent protein expression in the transfected NIH3T3 cells after G418 selection, and the highest percentage of labeled NIH3T3 cells was found in pLEGFP-N1 group. The population doubling time in pDsRed2-C1 (40.3+/-0.7 h) , PEGFP-N1 (39.6 +/- 0.6 h) and pLEGFP-N1 (36.5 +/- 0.7 h) groups was evidently longer than that in control (27.9 +/- 0.6 h, P < 0.01), with high adhesive rate in each group. CONCLUSION: The expression of fluorescent protein exhibited some inhibitory effect on the proliferation of NIH3T3 cells in vitro. Since the inhibitory effect by retroviral vector was weaker compared with eukaryotic vector, it should be the first choice for fluorescent protein labeling during cell transplantation.