Significant genes in response to low temperature in Fenneropenaeus chinensis screened through multiple transcriptome group comparisons.

Fenneropenaeus chinensis is a migratory marine species with a suitable growth at 18-30°C. To prolong breeding season and reduce mortality in winter, breeding new shrimp varieties with cold tolerance is essential. Genes upregulated and highly expressed at low temperature are reasonable candidate genetic markers for the breeding of cold tolerant strain variants. This study screened genes with these features by comparing multiple low-vs. normal-temperature transcriptome groups. The results showed that nine genes were upregulated and highly expressed at low temperature in more than seven of the nine comparison groups. Six of them were identified as genes encoding transcription factor ATF2, RNA recognition motif domain-containing protein, cytochrome b5-like protein, troponin C, tubulin alpha-1, and 18S/5.8S/28S rRNA, respectively. Cold-inducible upregulations of ATF2, cytochrome b5, and rRNAs were novel findings in this study. The other three novel genes were predicted to encode a membrane-bound extracellular protein and two lncRNAs. Four of the screened genes were verified by real time RT-PCR, and their expression levels were consistent with the sequencing results, demonstrating the accuracy of the transcriptome sequencing data. Function analysis showed that ATF2 might be the master transcription factor regulating the expressions of proteins involved in cellular responses to cold. The other genes played a role in events such as enhancing translation, increasing energy, inhibiting apoptosis, and preserving cell integrity. The expression features of these nine genes suggested that they were of great significance to the cold tolerance of shrimp.

Shorter Grazing Time and Supplementation Are Beneficial for Gastrointestinal Tract Development and Carcass Traits of Growing Lambs.

The effects of restricted grazing durations on the gastrointestinal development and carcass quality of growing lambs are poorly understood. In this study, 32 lambs were randomly assigned to four groups (n = 8, body weight = 21.86 kg) corresponding to 2, 4, 8 and 12 h of grazing per day. When off-pasture, all lambs were housed and fed concentrate and hay. When the grazing time decreased from 12 h to 2 h, the abomasum weight and large intestine length decreased (p = 0.019; p = 0.069). Compared to lambs grazed for 12 h, animals grazed for 2−4 h had a greater villus height and villus-to-crypt ratio in the duodenum, jejunum and ileum segments (p < 0.05); the 2 h lambs had superior carcass quality and a smaller diameter and area of the gluteus medium muscle fibers (p < 0.05), with no significant change after 4 h of grazing. The results indicated that shorter grazing times and supplementation were beneficial for the gastrointestinal tract development and carcass quality of growing lambs. Therefore, a better grazing management approach in Inner Mongolia could be to restrict the grazing of lambs to 4 h per day instead of grazing for more extended periods.

Comparison of Three Feeding Regimens on Blood Fatty Acids Metabolites of Wujumqin Sheep in Inner Mongolia.

Feeding regimens influence the fatty acid composition of animal-derived products. However, there is limited information on the effect of feeding regimens on the blood fatty acid composition and metabolic pathways of ruminant animals. In this study, 30 Wujumqin sheep were randomly assigned to three groups, PF (pasture feeding), PSF (pasture feeding plus corn supplementation) and BF (barn feeding), to examine the effects of feeding regimens on blood fatty acid composition and metabolic pathways through a metabolomic approach. The results showed that the BF sheep had increased serum n-6 polyunsaturated fatty acids levels, while the PF and PSF sheep had increased serum n-3 PUFA levels. Compared to the BF and PSF sheep that were fed ground corn, the PF sheep that only ate natural grass had up-regulated serum DHA levels. Meanwhile, blood metabolites from linoleic acid and arachidonic acid, including pro-inflammatory products (20-HETE, LTs, TX etc.) and anti-inflammatory products (LXB4, DHETs, HPETEs etc.) were elevated in the BF group. It was found that, compared to grazing, concentrate supplement feeding regimens, including either grazing plus supplements or feeding indoors, down-regulated blood n-3 PUFA biosynthesis and up-regulated the blood inflammatory compound metabolism by n-6 PUFA.

Host-guest modes and supramolecular frameworks of complexes of tetramethyl cucurbit[6]uril with 4-chloroaniline and 4,4'-diaminostilbene.

Since the first reportal on decamethylcucurbit[5]uril (Me10Q[5]) in 1992, substituted cucurbit[n]urils have attracted considerable research interest. In this study, the host-guest modes between the tetramethyl cucurbit[6]uril (TMeQ[6]) as a host and 4-chloroaniline and 4,4'-diaminostilbene (G1 and G2) as guests were investigated by single-crystal X-ray diffraction, NMR, ITC, UV-Vis spectrum, and MALDI-TOF mass spectrometry analyses. The experimental results showed that TMeQ[6] formed a 1 : 1 inclusion compound with G1, and the carbonyl portal of TMeQ[6] formed a 1 : 1 self-assembly with G2. Further, multi-dimensional supramolecular frameworks were formed driven by weak interaction forces in the system (hydrogen bonding, C-H⋯π interactions, ion-dipole interactions, and dipole-dipole interactions).

Thicker Shell Eggs with Enriched N-3 Polyunsaturated Fatty Acids and Lower Yolk Cholesterol Contents, as Affected by Dietary Nettle (Urtica cannabina) Supplementation in Laying Hens.

The nettle Urticacannabina (U.cannabina) is not only a functional edible food for humans but also a potential alternative feed resource for poultry, providing protein, fatty acids, vitamins, and minerals. The present study was conducted to evaluate the effects of dietary U.cannabina on the production of high-quality eggs with enriched n-3 polyunsaturated fatty acids (PUFA) and lower cholesterol contents. One hundred and twenty laying hens were assigned to three groups [control, 15% alfalfa meal supplementation (TRTA), and 15% U.cannabina supplementation (TRTU)]. The results showed that the feed intake, yolk color, and shell thickness were increased (p < 0.05) in the U.cannabina group. Moreover, cholesterol contents of the yolk and serum were lower in the U.cannabina group (p < 0.05). The total n-3 PUFA concentration in the yolk was increased while the ratio of n-6/n-3 was reduced in the U.cannabina group (p < 0.01). In conclusion, the dietary inclusion of U.cannabina increased shell thickness, yolk n-3 PUFA levels, and yolk color, and reduced cholesterol contents of the yolk and serum without any negative impacts on health or laying performance.

Tianxia120: A Multimodal Medical Data Collection Bioinformatic System for Proactive Health Management in Internet of Medical Things.

A digital medical health system named Tianxia120 that can provide patients and hospitals with "one-step service" is proposed in this paper. Evolving from the techniques of Internet of Medical Things (IoMT), medical dig data, and medical Artificial Intelligence, the system can systematically promote the change of service status between doctors and patients from "passive mode" to "proactive mode" and realize online service that is similar to offline medical treatment scenarios. The system consists of a patient terminal and a doctor terminal. They can perform online inquiry (through graphic, voice, telephone, video, etc.), electronic prescription, multiparameter self-diagnosis, cold chain logistics, medicine distribution, etc. The system can provide rich medical health information, medical tools browsing, and health care big data aggregation processing functions. Compared with the traditional medical system, this system has the characteristics of full function, rich data, and high security. It is expected to be applied to hospital applications and medical research to promote the construction and innovation of clinical medical disciplines.

A study on the self-assembly mode and supramolecular framework of complexes of cucurbit[6]urils and 1-(4-methoxyphenyl)piperazine.

Self-assembly between symmetrical dicyclohexyl-substituted cucurbit[6]uril (abbreviated as (CyH)2Q[6]) and cyclopentanocucurbit[6]uril (CyP6Q[6]) as hosts and 1-(4-methoxyphenyl)piperazine (MeOPP) as a guest molecule has been studied by means of single-crystal X-ray diffraction analysis, NMR, MALDI-TOF mass spectrometry, and other characterization methods. The experimental results showed that the self-assembly was driven by the formation of exclusion complexes by the cucurbit[n]uril and the guest, that is, supramolecular interaction between the negative charge of the cucurbit[n]uril portals and a coordination polymer guest. Complexes were formed between the positive charge of the cucurbit[n]uril outer wall and inorganic anions, thus generating self-assemblies with multi-dimensional and multi-level supramolecular frameworks.

Ginsenosides promote proliferation of granulosa cells from chicken prehierarchical follicles through PKC activation and up-regulated cyclin gene expression.

The effect of GS (ginsenosides) on proliferation of chicken GCs (granulosa cells) from prehierarchical SYF (small yellow follicles) was evaluated, and involvement of the PKC (protein kinase C) signalling pathway as well as mRNA expression of cyclins and CDK (cyclin-dependent kinase) were investigated. Whole SYF or GCs isolated from SYF were cultured in Medium 199 supplemented with 0.5% FCS (fetal calf serum). After 16 h, the cells were challenged with GS alone or in combination with PKC inhibitor H7 or activator PMA (phorbol 12-myristate 13-acetate) for 24 h in serum-free medium. Results showed that in both whole follicles and pure GCs monolayer culture system, GS (0.1-10 microg/ml) significantly increased the number of GCs in SYF in a dose-dependent manner, and this stimulatory effect was inhibited by H7, but enhanced by PMA. Meanwhile, the PCNA-LI (proliferating cell nuclear antigen labelling index) of GCs displayed similar changes with the cell number. Mechanism of GS action was further evaluated in cultured GCs separated from SYF. Western blot analysis showed that 10 microg/ml GS increased PKC translocation from cytoplasm to the plasma membrane of the GCs to become the active state. This effect was blocked by H7. Furthermore, GS up-regulated the expression of cyclin D1/CDK6 and cyclin E/CDK2 mRNAs in GCs; however, inhibition of PKC with H7 attenuated this stimulatory effect. These results indicated that GS could stimulate proliferation of chicken GCs through activated PKC-involved up-regulation of cyclin D1/CDK6 and cyclin E/CDK2 genes, subsequently promoting development of the chicken prehierarchical follicles.

Interactive actions of prostaglandin and epidermal growth factor to enhance proliferation of granulosa cells from chicken prehierarchical follicles.

The interactive actions of prostaglandin (PG) and epidermal growth factor (EGF) on proliferation of granulosa cells was investigated in prehierarchical small yellow follicles (SYF) of laying hens. The granulosa layers were dispersed into single cells by 12.5 microg/ml collagenase. After 16 h pre-incubation in 0.5% fetal calf serum-supplemented medium, the medium was replaced with serum-free medium. Immunocytochemical staining showed that granulosa cells expressed EGF and its receptor, and their expression was increased by PGE(1) (1-100 ng/ml) or forskolin (10(-7) to 10(-5)M) treatments. EGF receptor was also induced by its ligand EGF. The specific prostaglandin synthase inhibitors SC560 (for COX-1) and NS398 (for COX-2) suppressed EGF-stimulated increase of the granulosa cell number. Furthermore, the effect of EGF was confirmed by the immunocytochemical staining of the proliferating cell nuclear antigen in granulosa cells. Though EGF promoted the expression of both COX-1 and COX-2, the rescue experiment indicated that combined treatment of PGE(1) showed better rescuing effect on NS398 inhibition than SC560 at 10(-6)M, which implies COX-2 plays the predominant role in mediating EGF action. The above results indicate that reciprocal stimulation of intracellular PG and EGF production may enhance proliferation of granulosa cells, hence to facilitate development of chicken prehierarchical follicles.

Pro-proliferating effect of homologous somatic cells on chicken primordial germ cells.

Many studies demonstrated that chicken primordial germ cells (PGCs) could maintain undifferentiated state on mouse embryonic fibroblast feeders supplemented with growth factors and cytokines. However, the xenosupport systems may run risk of cross-transfer of animal pathogens from the other animal feeder, matrix to the PGCs, then influencing later transgenic technology. In this study, chicken PGCs were identified by alkaline phosphatase, stage-specific embryonic antigen-1 and Oct-4 immunocytochemical stainings. Three different homologous somatic cell feeder layers (chicken embryonic fibroblast feeder layer, CEF; embryonic skeletal myoblast feeder layer; follicular granulosa cell feeder layer) were used to support growth and proliferation of PGCs to find a better supporting culture system. In addition, the effects of fetal calf serum (FCS), leukemia inhibitory factor (LIF) and the combination of insulin, transferring and selenite (ITS) on PGC proliferation were compared. Results showed that CEF was the best supporter for PGC growth and proliferation, which was verified by 5-bromo-2'-deoxyuridine incorporation stain. FCS alone or in combination with LIF could significantly promote PGC proliferation in the presence of CEF in ITS medium. This study will contribute to providing a safer supporting system for chicken PGC amplification in vitro, and may be applied in transgenic chicken production and transplantation therapy.

Prostaglandin involvement in follicle-stimulating hormone-induced proliferation of granulosa cells from chicken prehierarchical follicles.

The aim of the present study was to evaluate the role of prostaglandin (PG) on proliferation of granulosa cells from prehierarchical small yellow follicles (SYF) of buff laying hens. The granulosa layers were separated by mechanic method and dispersed into single cells. After 16 h pre-incubation in 0.5% FCS medium, the medium was replaced with serum-free medium, which was supplemented with 10 microg/ml insulin, 5 microg/ml transferrin and 3 x 10(-8)M selenite. Cells were challenged with PGE1 and FSH for 24 h and then assessed for proliferation. The results showed that PGE(1) (0.1-10 ng/ml) had a similar proliferating effect as FSH on granulosa cells, and these stimulating effects were restrained by the PGE receptor antagonist SC19220 at 10(-7) to 10(-5)M. Prostaglandin synthase antagonist indomethacin (10(-7) to 10(-5)M) suppressed FSH-induced increase in the number of granulosa cells in a dose-dependent manner. Downstream activation of protein kinase A by forskolin-activated adenylate cyclase resulted in elevated proliferation of granulosa cells, an effect unobserved by phorbol-12-myristrate-13-acetate-activated protein kinase C. In addition, PGE1-stimulated proliferation of granulosa cells was hindered by H89 (PKA inhibitor) but not by H7 (PKC inhibitor). Furthermore, the proliferating cell nuclear antigen labeling index (PCNA-LI) of granulosa cells displayed similar changes with the number of cells. These results indicated that PGE1 promoted the proliferation of granulosa cells from SYF and was also involved in mediating FSH-stimulated intracellular PKA signal transduction.