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Currently, feed enzymes are primarily obtained through fermentation of fungi, bacteria, and other microorganisms. Although the manufacturing technology for feed enzymes has evolved rapidly, the activities of these enzymes decline during the granulating process and the cost of application has increased over time. An alternative approach is the use of genetically modified plants containing complex feed enzymes for direct utilization in animal feedstuff. We co-expressed three commonly used feed enzymes (phytase, β-glucanase, and xylanase) in barley seeds using the Agrobacterium-mediated transformation method and generated a new barley germplasm. The results showed that these enzymes were stable and had no effect on the development of the seeds. Supplementation of the basal diet of laying hens with only 8% of enzyme-containing seeds decreased the quantities of indigestible carbohydrates, improved the availability of phosphorus, and reduced the impact of animal production on the environment to an extent similar to directly adding exogenous enzymes to the feed. Feeding enzyme-containing seeds to layers significantly increased the strength of the eggshell and the weight of the eggs by 10.0%-11.3% and 5.6%-7.7%, respectively. The intestinal microbiota obtained from layers fed with enzyme-containing seeds was altered compared to controls and was dominated by Alispes and Rikenella. Therefore, the transgenic barley seeds produced in this study can be used as an ideal feedstuff for use in animal feed.

The (co)variance components and corresponding phenotypic and genetic parameters for growth traits and wool traits of economic importance were estimated in the Alpine Merino sheep population maintained at Gansu Provincial Sheep Breeding Technology Extension Station in northwestern China. Data from a maximum of 49,474 animals sired by 526 rams and born from 22,531 ewes over 20 years from 2000 to 2019 were used in this study. Birth type, age of dam, birth year, sex and/or management group, and age at measurement were initially fitted as fixed effects in an animal model with various random effects. Genetic groups were defined for all animals by the sire breed and breed genotype interacted with dam-strain flocks and were fitted as one of the random effects. Analyses were conducted using a residual maximum likelihood procedure (ASReml). Seven different animal models were fitted for all traits, and the most appropriate model with relevant random effects was selected through log-likelihood ratio testing. After identifying the appropriate model through single-trait analysis, bivariate analyses were used to obtain the phenotypic and genetic correlations among the traits. The estimates of additive direct heritability for birth weight (BWT), weaning weight (WWT), preweaning growth rate (prwADG), postweaning growth rate (powADG), yearling body weight (YWT), average fibre diameter (AFD), greasy fleece weight (GFW), clean fleece weight (CFW), yield (YLD), yearling wool staple length (YSL), coefficient of variation of average fibre diameter (FDcv) and wool visual fineness counts (VFC) were 0.30, 0.18, 0.18, 0.20, 0.29, 0.20, 0.19, 0.20, 0.35, 0.19, 0.16 and 0.13, respectively, with standard errors ranging from 0.02 to 0.05. The corresponding ratios of genetic group variance to additive genetic variance were significant and, respectively, 0.35, 0.80, 0.62, 0.26, 0.13, 1.06, 0.38, 0.64, 0.09, 0.12, 0.06 and 0.58. These results suggest for these traits that there is potential to exploit both the additive genetic variation and between genetic group variation although for most traits the between group variation was smaller than the variation within groups. Favourable genetic correlations were found among the growth traits, and between growth traits and fleece production traits, and among wool traits GFW, CFW, YSL and YLD. This study provides the required estimates of genetic parameters of both growth and wool traits of the new breed for the design of more effective breeding programmes.

One dimensional (1D) organic-inorganic halide hybrid perovskite has the advantages of excellent organic cation modifiability and diversity of inorganic framework structures, which cannot be ignored in the development of multi-functional phase transition materials in photoelectric and photovoltaic devices. Here, we have successfully modified and synthesized an organic-inorganic hybrid perovskite photoelectric multifunctional phase transition material: [C7H13ONCH2F] PbBr3 (1). The synergistic effect of the order double disorder transition of organic cations and the change of the degree of distortion of the inorganic framework leads to its high temperature reversible phase transition point of Tc=374 K/346 K and its ultra-low loss high-quality dielectric switch response. Through in-depth research and calculation, compound 1 also has excellent semiconductor characteristics with a band gap of 3.06 eV and the photoluminescence characteristics of STE broadband emission. Undoubtedly, this modification strategy provides a new choice for the research field of organic-inorganic hybrid perovskite reversible phase transition photoelectric multifunctional materials with rich coupling properties.

The aim of this study was to reveal the effect of let‐7b on osteoporosis (OP). Synthetic let‐7b mimics or inhibitors were transfected into MC3T3‐E1 cells. The expression of let‐7b in MC3T3‐E1 and its effect on cell viability, apoptosis, and the apoptosis‐related proteins (Bcl‐2, Bax, and cleaved caspase‐9) were tested by CCK‐8 assay, flow cytometry and Western blot, severally. The osteogenic differentiation markers (Runx2 and Osterix) and Wnt/β‐catenin pathway related markers (β‐catenin and C‐myc) were detected by qRT‐PCR and Western blot. The relationships between let‐7b and cyclin D1 (CCND1) were confirmed by luciferase reporter assay. The differentiation and mineralization of MC3T3‐E1 cells were analyzed by alkaline phosphatase (ALP) activity assay and alizarin red staining. The outcomes indicated that overexpression/ablation of let‐7b repressed/facilitated MC3T3‐E1 cell viability and accelerated/suppressed MC3T3‐E1 cell apoptosis. Besides, a remarkable decrease/augment of Bcl‐2 protein expression and the distinct fortify/reduction of Bax and cleaved caspase‐9 expression levels were observed in let‐7b mimics/inhibitors group in MC3T3‐E1 cells. Moreover, we discovered that let‐7b overexpression/ablation retrained/facilitated the mRNA and protein expression of Runx2 and Osterix. It was confirmed that CCND1 was a downstream target of let‐7b and was negatively modulated by let‐7b. In addition, high‐expression/deficiency of let‐7b inhibited/increased the expression levels of β‐catenin and C‐myc in MC3T3‐E1 cells. Taken together, our study revealed that let‐7b overexpression/depletion repressed/accelerated MC3T3‐E1 cell proliferation, differentiation, and mineralization while promoted/suppressed MC3T3‐E1 cell apoptosis through targeting CCND1, which might be adjusted by Wnt/β‐catenin pathway. Our findings might offer a basis for developing novel targets for OP treatment.

It was found that the expression level of miR-147a was significantly increased and the pathway of PI3K/AKT was dramatically inhibited after radiation. In view of the relationship between miRNA and target genes, we put forward the question, what is the relationship between PI3K/AKT and miR-147a? In order to find the answer to the question, we used bioinformatics techniques to analyze the relationship between miR- 147 (a or b) and PI3K/AKT signaling pathway. miR-147a overexpression plasmid and PDPK1 3′UTR luciferase reporter gene plasmid were constructed. Dual luciferase reporter gene system validation experiments were carried out on miR-147a and PDPK1 relationship. The verification experiments were also carried out. Bioinformatics analysis showed that there is a miR-147a binding site in the non-coding region (3′UTR) of PDPK1. In the experimental groups transfected with wild type PDPK1 gene of 3′UTR plasmid, the luciferase activity decreased (or increased) significantly in miR-147a (or inhibitor) group compared with miR-NC (or anti-miR-NC); There was no significant difference between the miR-147a group (or inhibitor) and the miR-NC group (or anti- miR-NC) in the transfection of PDPK1–3′UTR-Mut gene vector. PDPK1 was a target gene for direct regulation of miR-147a downstream. Verifying test results showed that the expression of PDPK1 mRNA and protein was reduced after overexpression of miR-147a, which was up-regulated after silencing miR-147a in TC, and V79 cells. These results suggest that miR-147a could be involved in the regulation of PDPK1 transcription by binding to the target site in PDPK1 mRNA 3′UTR, and then regulated AKT.

The aim of this study is to examine the burden of family caregivers of patients with retinoblastoma in China and the relationships between depression, caregiver burden and social support. A descriptive and correlational survey was conducted with 117 Chinese family caregivers of outpatient patients with retinoblastoma from the Department of Ophthalmology of a tertiary hospital in Shanghai, China. Family caregivers of outpatient patients with retinoblastoma were asked to respond to four questionnaires including sociodemographic questionnaire, Becker Depression Inventory, Caregiver Burden Inventory and Social Support Rating Scale. The incidence of depression in this study was 51.3%; the average score for social support indicated moderate social support available to the caregivers, although their level of caregiver burden was heavy. Depression scores were significantly positively correlated with caregiver burden scores and significantly negatively correlated with the social support scores. Heavy caregiver burden was associated with lower monthly income, low subjective social support and less use of social support.

Pulmonary arterial hypertension (PAH) is a life-threatening disease characterized by progressive pulmonary arterial remodeling and right ventricular failure. Despite recent advances in pathophysiological mechanism exploration and new therapeutic approaches, PAH remains a challenging condition. In this study, we investigated the roles of the peptide fragments from proadrenomedullin (proADM) such as adrenomedullin (ADM), adrenotensin (ADT), and proadrenomedullin N-terminal 20 peptide (PAMP) during pulmonary remodeling caused by high pulmonary blood flow, and probed the possible involvement of mitogen-activated protein kinase (MAPK) signal transduction pathways. Sixteen rat models of PAH were artificially established by surgically connecting the left common carotid artery to the external jugular vein. We subcutaneously injected an extracellular signal-regulated protein kinase (ERK1/2) inhibitor, PD98059, in eight rats, treated another eight rats with an equal volume of saline. Eight rats without connections served as the control group. We observed that mRNA expression levels of ADM, stress-activated protein kinase (SAPK), and ERK1/2 were significantly elevated in the shunted rats; furthermore, ERK1/2 levels were significantly inhibited by PD98059. Protein levels of ADM, PAMP, p-SAPK, and p-ERK1/2 were significantly higher ADT was lower, and p-p38 remained unchanged in the rat models compared with the controls. However, the protein expression of both ADM and p-ERK1/2 was significantly inhibited by PD98059. Our results suggest that levels of ADM, ADT, and PAMP respond to pulmonary remodeling, and that activation of the SAPK and ERK1/2 signaling pathways is involved in pulmonary hypertension and artery remodeling caused by high pulmonary blood flow.

Summary Background Baroreflex gain increase up closely to adult level during initial postnatal weeks, and any interruption within this period will increase the risk of cardiovascular problems in later of life span. We hypothesize that this short period after birth might be critical for postnatal development of vagal ganglion neurons (VGNs). Methods To evaluate neuroexcitability evidenced by discharge profiles and coordinate changes, ion currents were collected from identified A- and C-type VGNs at different developmental stages using whole-cell patch clamping. Results C-type VGNs underwent significant age-dependent transition from single action potential (AP) to repetitive discharge. The coordinate changes between TTX-S and TTX-R Na+ currents were also confirmed and well simulated by computer modeling. Although 4-AP or iberiotoxin age dependently increased firing frequency, AP duration was prolonged in an opposite fashion, which paralleled well with postnatal changes in 4-AP- and iberiotoxin-sensitive K+ current activity, whereas less developmental changes were verified in A-types. Conclusion These data demonstrate for the first time that the neuroexcitability of C-type VGNs increases significantly compared with A-types within initial postnatal weeks evidenced by AP discharge profiles and coordinate ion channel changes, which explain, at least in part, that initial postnatal weeks may be crucial for ontogenesis in visceral afferent reflex function.

The mechanisms through which bone marrow adipocytes might influence differentiation and function of osteoblasts are not completely understood. To investigate the direct effects of bone marrow fat cells on osteoblast function, an ex vivo co-culture system was utilized comprising either primary fat cells or differentiated 3T3-L1 adipocytes and osteoblastic cells on transwells. In co-culture, both adipocytes and osteoblastic cells were differentiated into adipocytes or osteoblasts, respectively, before culturing on transwells. Co-culture with either primary fat cells or fully differentiated 3T3-L1 adipocytes significantly decreased mRNA and protein expression of runt-related transcription factor 2 (Runx2) in osteoblastic cells. An increase in mRNA and protein expression of peroxisome proliferator-activated receptor γ (PPARγ) occurred concomitantly with the reduction of Runx2 expression. Adiponectin concentration was increased in the media by co-culture. In addition, co-culture with conditioned media from fat cells increased PPARγ promoter activity and decreased Runx2 promoter activity. Knockdown of PPARγ or adiponectin receptor 1 in osteoblastic cells by siRNA prevented the down-regulation of mRNA expression of Runx2 in osteoblastic cells cultured with fully differentiated 3T3-L1 cells. Furthermore, co-transfection with PPARγ decreased Runx2 promoter activity. A marker of osteogenesis, alkaline phosphatase activity in osteoblastic cells was significantly decreased by co-culture. Annexin V/propidium iodide staining showed that co-culture did not induce apoptosis in osteoblastic cells. Thus, we conclude that adipocytes modulate key metabolic functions of osteoblasts through the release of secretory products. PPARγ plays a key role in mediating the effects of adipocytes on osteoblasts.

Effective delivery of therapeutic genes or small molecular drugs into macrophages is important for cell based immune therapy, but it remains a challenge due to the intracellular reactive oxygen species and endosomal degradation of therapeutics inside immune cells. In this report, the star-like amphiphilic biocompatible β-cyclodextrin-graft-(poly(e-caprolactone)-block-poly(2-(dimethylamino) ethyl methacrylate)x (β-CD-g-(PCL-b-PDMAEMA)x ) copolymer, consisting of a biocompatible cyclodextrin core, hydrophobic poly(e-caprolactone) PCL segments and hydrophilic PDMAEMA blocks with positive charge, is optimized to achieve high efficiency gene transfection with enhanced stability, due to the micelle formation by hydrophobic PCL segments. In comparison with lipofetamine, a currently popular nonviral gene carrier, β-CD-g-(PCL-b-PDMAEMA)x copolymer, shows better transfection efficiency of plasmid desoxyribose nucleic acid in RAW264.7 macrophages. More interestingly, this delivery platform by β-CD-g-(PCL-b-PDMAEMA)x not only shows low toxicity but also better dexamethasone delivery efficiency, which might indicate its great potential in immunotherapy.

Objectives To provide a new, reliable noninvasive method for fetal sex determination. Methods Fetal sex was detected in 32 early pregnant women by identifying the amelogenin gene in maternal plasma using nested PCR analysis. First, the 122/128 bp of X-Y homologous region containing 6 bp deletions in the intron 3 of amelogenin gene in X chromosome was amplified, and then the nested PCR was carried out, whose 3′ end of the upstream primer is just located in the deletion region. The fetus was male or female, depending on whether it had the 89-bp nested PCR product or not. Results The 89 bp of nested PCR product was detected in 19 plasma samples obtained from pregnant women, deducing they bear the male fetus and the remaining pregnant women bear female. When compared with the birth outcome, two samples were pseudo-positive. The coincidence was 93.8%. This method had high sensitivity that even trace amount of target fetal DNA (10 pg) could be detected. Conclusions This conventional nested PCR analysis of amelogenin gene promises to be a reliable method for noninvasive fetal sex determination at early pregnancy using maternal plasma DNA. Copyright © 2005 John Wiley & Sons, Ltd.

Skeletal sphalerite with stellar, cruciform and snowflake-like (or cauliflower-like) shapes included in pyrite is widely found in the Dajing tin-polymetallic deposit. It occurs only in chalcopyrite-pyrite mineralization stage. The compositions of all sphalerites in the chalcopyrite-pyrite stage are characterized by high Cu content (3.9 - 7.0 wt% with a mean of 5.4 wt%), while the skeletal crystal sphalerite has higher zinc and cadmium contents, and lower copper and iron contents, compared with other sphalerites of the same stage. The skeletal crystal sphalerite in pyrite is possibly generated by exsolution.

The southern part of the Da Hinggan Mountains is the only tin-polymetallic concentration area in northern China. Based on ore-forming element assemblages, three metallogenic series, the Sn(W), paragenetic Sn–polymetallic, and poly-metallic series, are recognized. The Sn(W) series, consisting of greisen, skarn and vein types, is associated with ilmenite–series, potassic feldspar (Kf)–granites. The polymetallic Pb–Zn–Cu series with porphyry, skarn and vein types, is related to magnetite–series, granodiorite – monzonite. On the contrary, the paragenetic Sn-polymetallic series mineralization is associated with the coexisting igneous activities of Kf-granite and granodiorite–monzonite, and it is suggested that the paragenetic Sn-polymetallic series is caused by the superimposed mineralization with tin from ilmenite-series magma and polymetallic elements from magnetite-series magma–hydrothermal system. All the three series possess similar metallogenic age, concentrating on J3–K1 (130–150 Ma). The melting of high maturity and tin-rich Xilinhot Proterozoic complex (micro–massif) during Mesozoic period, could generate the ilmenite–series, tin-bearing felsic magma, and cause the tin mineralization in the southern part of the Da Hinggan Mountains.

The skarns and genesis were studied of the Huanggang Fe-Sn deposit and the nearby Sumugou Zn-Pb deposit in Inner Mongolia, China. In the Huanggang mine, Nos. 1 to 4 Fe ore bodies are arranged along a calcareous horizon from proximal to distal in this order to a granite intrusion named Luotuochangliang, while Sn ore body is situated near another granite intrusion named 204. According to the distance from the granitic intrusions, mineral assemblages in skarns are systematically changed. Garnet is the most predominant skarn mineral throughout the deposit. Hastingsitic amphiboles, however, predominate in the proximal skarns. Fluorite is common in the proximal skarns, while instead calcite is common in the distal skarns. Chlorite is characteristically present only in No. 3 ore body, and chlorite geothermometry gives near 300C for the mineralization of later stage. When garnet crystal shows zonal structure, isotropic andraditic garnet occupies the core, and is surrounded with anisotropic less-andraditic garnet. The presence of white skarn along the boundary between main skarns and host sedimentary rocks confirms relatively reducing environment prevailing as a whole in the studied area. However, the compositional relation between coexisting garnet and clinopyroxene demonstrates that relatively oxidizing condition was achieved for garnet skarn and magnetite ore in the distal, Nos. 2 to 4 Fe ore bodies and Sumugou deposit, compared to that for garnet skarn in the proximal, No. 1 and Sn ore bodies. Preliminary study on the tin content of garnets in the studied area revealed a certain degree of contribution brought from granitic intrusives since the early stage of skarn formation, irrespective of proximal or distal. Oxygen isotope study on garnet, magnetite, quartz and skarn calcite, as well as hydrogen isotope study on hastingsitic amphibole, demonstrates mainly meteoric water origin for the skarn– and ore-forming solutions. The occurrence of Sn, W, Mo and F minerals indicates that those elements were mainly supplied to the deposit later than the formation of skarns and iron ores, overlapping to them. These constraints allow to delineate the formation model of the deposit as follows (Fig. 10): At the time of late Jurassic to early Cretaceous, felsic activity occurred in this region as a part of Yanshanian magmatism, and formed granitic intrusions as well as thick volcanic piles on the surface. The circulation of meteoric water was provoked by the heat brought by the intrusions. By this circulation, much amount of iron was extracted from andesites of the Dashizhai Formation, and precipitated as skarns and magnetite ores along calcareous horizons near the bottom of the Huanggangliang Formation. Subsequently, volatile-rich fluids with Sn, W and Mo were expelled from the solidifying granitic magmas, and precipitated these metals in the pre-existing skarns and ores.

Dajing is a large-scale tin–polymetallic ore deposit in north China with Sn, Cu, Pb, Zn, Ag, and minor elements such as Co and In. The deposit is controlled by a passage-host structure system. Two groups of NE direction faults such as F2 and F1 are characterized by sinistral and multiphase activities, and the syn-ore NE faults with their derivative faults nearly in N-S direction constitute an important structure system as channelway for ore fluid. Around F2 fault (mineralization center), metallic elements display horizontal zoning outwards from Sn, Cu to Cu, Sn, Pb, Zn, Ag and to Pb, Zn, Ag. Controlled by sinistral pull-apart vein system, the WNW direction host structure yielded the ore vein to “W” shape on horizontal plan while to staircase shape on vertical profile. Multiperiodic activities of the host structures system lead to multistage of ore mineralization. Four mineralization stages are recognized in the main mineralization, i.e. 1) cassiterite–arsenopyrite–quartz stage, 2) cassiterite-sulfide (or Cu–poly–metallic mineralization) stage, 3) massive pyrite stage, and 4) galena-sphalerite stage. Besides, fluid activities are identified here at the pre-mineral stage of the formation of quartz veins and sheared deformation, and at the post-mineral stage of quartz–calcite–fluorite deposition. The earlier tin-rich ore solution raised from NE faults and its nearly N-S derivatives filled the NW-WNW direction fractures, and formed tin ore veins. Subsequent mineralization of Cu-polymetallic stage, pyrite stage and galena-sphalerite stage, overlapping on the same fracture system, sequentially expanded outwards from the fractures, and produced different kinds of ore types and mineralization zoning.

A new strategy for the synthesis of tetrahydroimidazo[1,2-a]pyridines and unusual tetrahydrobenzo[b]imidazo[1,2,3-ij][1,8]naphthyridines has been successfully developed by cascade reactions including Knoevenagel condensation, aza–ene reaction, imine–enamine tautomerization, cyclocondensation/oxidation, and intramolecular SNAr of precursors 2-(2-chloroaroyl)methyleneimidazolidines as new heterocyclic ketene aminals (HKA), which represent a class of polyfunctional scaffolds with four active reaction sites with aromatic aldehydes and malononitrile or ethyl 2-cyanoacetate under mild conditions. In this domino reaction, nine different active sites are involved, and two C–C bonds, two C–N bonds, and two new rings are constructed with all reactants efficiently utilized in the chemical transformation.