ALK Family Inhibitor A83-01 Promotes the Proliferation of Mouse Male Germline Stem Cells （mGSCs） Under Serum- and Feeder-Free Conditions

A83-01 is a selective inhibitor of the TGF-β type I receptor ALK,which inhibits the TGF-β-induced epithelial-to-mesenchymal transition(EMT) via the inhibition of Smad2 phosphorylation.Previous studies have showed that A83-01 promoted somatic cellular reprogramming significantly.Male germline stem cells(mGSCs),as an alternative resource of pluripotent stem cells derived adult testis,have promising valuable in clinic medicine and regeneration,however,the derivation of mGSCs was complex and difficult.What the role A83-01 plays in promoting the proliferation of mGSCs is still unknown.In this study,combined with A83-01 and knockout serum replacement(KSR) medium,we obtained a relatively feeder-and serum-free system for mGSCs culturing in vitro and the optimal concentration of A83-01 was 0.25 μmol L-1.After continuous culturing,the proliferation efficiency of undifferentiated mGSCs and differentiation capacity of mGSC were examined as well.Results showed that,A83-01 dramatically increased the number of mGSCs and AP positive colonies,and the mitosis index according to the BrdU assay.A83-01 could also increase the expression of pluripotent markers including Oct4,Klf4,Nanog and c-Myc,analyzed byreal-time quantative PCR.mGSCs cultured in the optimal feeder-and serum-free system combined with A83-01 could form embryoid bodies(EBs),which consisted of three embryonic layers detected by immunofluorescence and RT-PCR.Remarkably,the results demonstrated 0.25 μmol L-1A83-01 could promote the proliferation of mouse mGSC colonies and maintain their undifferentiated status under feeder-and serum-free systems.     

The environmental prospects of cultured meat in China

To deal with concerns in China about environmental degradation and a growth in population accompanied by increased consumption of livestock products, a meat alternative is required. This study compared the environmental impacts of producing different protein sources for nutrition, including crops, livestock products, and cultured meat. The results showed that cultured meat has the lowest land use per unit of protein and unit of human digestible energy. China’s crops have the lowest energy use and greenhouse gas(GHG) emissions per unit of energy and protein. The energy use in cultured meat production is slightly higher than that of current pork production in China, whereas GHG emissions are lower. It is concluded that the overall impact of replacing livestock products with cultured meat would be beneficial for China’s environment and would potentially improve food security because less land is needed to produce the same amount of protein and energy.     

雌二醇对体外培养绒山羊初级毛囊的影响(英文)

[Objective] The aim of this study was to preliminarily explore the effects of estradiol on morphology and growth of cashmere goat primary hair follicles. [Method] Cashmere goat primary hair follicles were cultured in serum-free Williams E media supplemented with different doses of 17 β-E2 (0, 0.1, 1.0, 10.0, 100.0 nmol/L), and their growth rates and morphological changes were observed. [Result] The growth rate of 0.1 nmol/L 17 β-E2 group was quite comparable with that of the control group(0 nmol/L), but the 17 β-E2 with concentrations of 1.0, 10.0 and 100.0 nmol/L displayed different degrees of inhibition on the growth of hair follicles. Different morphological changes of hair follicles could also be discovered in different concentration treatments. [Conclusion] The study laid a certain foundation for exploring the regulation mechanism of estrogen on growth of cashmere goat hair follicles.     

Revenue Sharing in Dairy Industry Supply Chain-A Case Study of Hohhot,China

Dairy industry has become an increasingly important enterprise in China as people＇s dietary preferences and composition have changed dramatically with rapid economic development in the past several decades.A number of problems,however,exist in China＇s relatively young dairy industry,including the imbalanced allocation of profits throughout the dairy supply chain.One of the root causes of the melamine infant powered milk scandal in 2008 was the unfair profit allocation mechanism in dairy supply chain.The revenue sharing contract approach has proven to be effective in generating market shares and total profits.In this study,we apply the three-stage revenue sharing contract model of Giannoccaro and Pontrandolfo（2004） in an analysis of dairy supply chain to explore its problems in profit allocation and possible solutions to them.The analysis was conducted by a case study of Hohhot,often called as ＂milk capital of China＂.Our results show that the current profit distribution in the dairy supply chain is not balanced：the supermarket＇s profitfarmer＇s profitmanufacturer＇s profit.Under the revenue sharing contract setting,the dairy industry＇s total profit increased by 12.49%.By exploring different parameters in the revenue sharing contract model,we have found that a win-win situation can be created among all the members of the supply chain.In dairy supply chain,the ratio of the revenue reserved for the supermarket itself is equal or greater than 47% and the ratio of the revenue reserved for the manufacturer itself is between 46.4 and 50.2%.The values of the parameters that generate a sustainable or win-win situation are related to the bargaining position in the dairy supply chain.The revenue sharing contract has proven to be effective and desirable by all the dairy chain partners in dairy supply chain.The results of this study provide relevant information for improving the dairy supply chain structure and the revenue sharing contract model can be applied to other industries,sectors and regions.     

Polymorphism of Exon 2 of FSHβ Gene and Its Relationship with Reproduction Performance in Two Goat Breeds

The objectives of the present study were to detect the polymorphism in follicle stimulating hormone beta （FSHβ） gene and investigate the relationship between FSHβ gene and high prolificacy in goats.According to the sequence of ovine FSHβ gene,two pairs of primers were designed to detect single nucleotide polymorphism of exon 2 of FSHβ gene in two goat breeds,Xinong Saanen dairy goat and Boer goat,by PCR-SSCP.The least square mean and genetic variance of different genotypes at polymorphic locus were analyzed,and the selection reaction was forecasted by genetic correlation between marker locus and genetic variance of litter size traits.There was one polymorphic locus occurring in exon 2 （P2） with three genotypes （EE,EF and FF）.In Xinong Saanen dairy goat,EE genotype had significantly higher （P 〈 0.05） litter size than EF and FF genotype in the first to fourth parity and average parity;EF genotype had significantly higher （P 〈 0.05） litter size than FF genotype in the second and average parity.In Boer goat,EE genotype had significantly higher （P 〈 0.01） litter size than EF and FF genotype in the second to fourth parity and average parity;EF genotype had significantly higher （P 〈 0.05） litter size than FF genotype in average parity.In Xinong Saanen dairy and Boer goat,the heredity of litter size was mainly influenced by genetic additive effect and the EE genotype is a favorable marker genotype for litter size in these two breeds.     

Segmentation of Somatic Cells in Goat Milk Using Color Space CIELAB

Somatic cell counts （SCCs） levels indicate the occurrence of infections in goat udders and are related to the productivity of goat milk, cheese and yoghurt. This work presents a segmentation method for counting somatic cells in goat milk images, intending to detect an infection known as mastiffs, which is the major cause of loss in dairy farming. The image segmentation procedure is devised by using the lab color space and the watershed transform. A large number of samples under variable preparation conditions are treated with the proposed method. A comparison between manual and the proposed technique is presented. Promising results indicates that video-microscopy systems may be employed to develop automated SCC for goat milk.     

Effect of sucrose on cryopreservation of pig spermatogonial stem cells

Sucrose is known to play an important role in the cryopreservation of sperm and female gonads; however, its effect on the cryopreservation of pig spermatogonial stem cells(p SSCs) has not been tested. The aim of this work was to study the effect of sucrose during p SSC cryopreservation and to find the most effective concentration in freezing medium. p SSCs were cryopreserved with freezing media containing different concentrations of sucrose(70, 140, 210, and 280 mmol L~(–1)) and a control group without sucrose. The survival rates, plasma membrane integrity, and mitochondrial membrane potential of thawed cells were detected by trypan blue(TB) staining, SYBR-14/propidium iodide(PI) dual staining, and JC-1 staining, respectively. All the staining results showed an obvious increase in cell survival in the sucrose-treated groups as compared to that in the control group, with the exception of 280 mmol L~(–1) sucrose. Moreover, the 210 mmol L~(–1) sucrose group yielded the highest survival rate among all the groups(P0.05). The results of SYBR-14/PI dual staining and JC-1 staining were consistent with those of TB staining as above described. Quantitative real-time PCR(q RT-PCR) indicated that the m RNA levels of three apoptosis-promoting genes(BAX, APAF1 and CASPASE9) were significantly higher in thawed cells than in cells before freezing(P0.05). Moreover, the mR NA level of one anti-apoptotic gene(XIAP) was significantly lower in thawed cells than in cells before freezing(P0.05). When comparing the m RNA expression of apoptosis-related genes in thawed cells, the m RNA level of the anti-apoptotic genes in the control group was significantly lower than that in the sucrose-treated groups(P0.05). Western blot analyses showed that the expression levels of cleaved CASPASE9, CASPASE3 and PARP-1 in the sucrose-treated groups were lower than those in the control group and were the lowest in the 210 mmol L~(–1) sucrose group. Both q RT-PCR and Western blot analyses suggested that sucrose inhibited cell apoptosis during freezing and thawing. Briefly, sucrose promoted p SSCs survival after freezing and thawing, especially at a concentration of 210 mmol L~(–1), which possibly assisted p SSC dehydration and inhibited cell apoptosis. These findings hold great promise for further studies of the regulatory mechanism of proliferation and differentiation of p SSCs.     

Conophylline Promotes the Proliferation of Immortalized Mesenchymal Stem Cells Derived from Fetal Porcine Pancreas (iPMSCs)

Conophylline, is a bis (indole) alkaloid consisting of two pentacyclic aspidosperma skeletons, isolated from Tabernaemontana divaricata, which has been found to induce β-cell differentiation in rat pancreatic acinar carcinoma cells and in cultured rat pancreatic tissue. However, the precise role of conophylline in the growth and survival of immortalized pancreatic mesenchymal stem cells (iPMSCs) derived from fetal porcine pancreas were not understood at present. To determine whether this molecule is involved in controlling the proliferation of iPMSCs, we examined the effects of conophylline on iPMSCs. We found that conophylline can robustly stimulate iPMSCs proliferation, even promote their potential differentiation into islet-like clusters analyzed by cell counting, morphology, RT-PCR and real-time PCR, Western blotting, glucose-stimulated insulin release and insulin content analysis. The effects of conophylline were inhibited by LY294002, which is the inhibitor of the PI3K pathway. These results suggest that conophylline plays a key role in the regulation of cell mass proliferation, maintenance of the undifferentiated state of iPMSCs and also promotes iPMSCs differentiated into insulin-producing cells.     

Effects of Different Dietary Cation-Anion Difference on Fiber Degradation in Rumen of Laoshan Dairy Goats

The experiment was conducted to determine effects of different dietary cation-anion difference（DCAD） in diets on ruminal fluid pH and fiber degradation in rumen of Laoshan dairy goats. A 4×4 latin square design was adopted. DCAD in different groups was 0, 50, 100, 200 mEq·kg^-1 of DM, respectively. The results of ruminal pH indicated that different DCAD could significantly influence the ruminal pH （P〈0.05） and ruminal fluid pH increased as DCAD increased from 0 to 200 mEq·kg^-1 of DM at different sampling time-points. There was no effect of DCAD on carboxymethyl cellulase in ruminal fluid at 4 h and 8 h postfeeding （P〉0.05）. Degradation ofNDF, ADF, CF peaked at a DCAD of 100 mEq·kg^-1 of DM. It could be concluded that DCAD of 100 mEq·kg^-1 of DM was advantage to non-pregnancy, non-lactication Laoshan dairy goat.     

Galactopoietic Activity of Dibutyl Phthalate Isolated from Vaccaria segetalis

A galactopoietic compound, identified as dibutyl phthalate（DBP）, was isolated from Vaccaria segetalis. The activity of DBP on lactation ability of dairy cow mammary gland epithelial cells（DCMECs） cultured in vitro and dairy cow was evaluated. Results showed that DBP could promote cell viability, proliferation ability, lactose and β-casein secretion of DCMECs, which could also raise the milk yields of dairy cows significantly.     

Construction of Antibacterial Peptide CecropinB Eukaryotic Recombinant Vector and Its Expression in Dairy Goat Mammary Gland Epithelial Cells

To investigate the expression of antibacterial peptide CecropinB cDNA in dairy goat mammary gland epithelial cells,the CecropinB gene was cloned and was inserted into a eukaryotic vector pECFP-C1 to construct the recombinant plasmid pECFP-B by genetic engineering technique.Recombinant plasmid pECFP-B was transfected into dairy goat mammary gland epithelial to detect the bactericidal activity of CecropinB.The expression of CecropinB was also detected.The result of RT-PCR demonstrated CecropinB gene was expressed in transfected cells.CecropinB recombinant plasmid DNA was injected into udders and CecropinB was expressed in mammary gland,exhibiting bactericidal activity to Staphylococcus aureus in vivo experiments.     

Genome Array on Differentially Expressed Genes of Skin Tissue in Cashmere Goat at Early Anagen of Cashmere Growth Cycle Using DNA Microarray

In order to study the molecular mechanism involved in cashmere regeneration, this study investigated the gene expression profile of skin tissue at various stages of the cashmere growth cycle and screen differentially expressed genes at proangen in 10 cashmere goats at 2 years of age using agilent sheep oligo microarray. Significance analysis of microarray （SAM） methods was used to identify the differentially expressed genes, Hierarchical clustering was performed to clarify these genes in association with different cashmere growth stages, and GO （Gene ontology） and the pathway analyses were con-ducted by a free web-based Molecular Annotation System3.0 （MAS 3.0）. Approximately 10200 probe sets were detected in skin tissue of 2-yr-old cashmere goat. After SAM analysis of the microarray data, totally 417 genes were shown to be differentially expressed at different cashmere growth stages, and 24 genes are significantly up-regulated （21） or down-regulated （3） at proangen concurrently compared to angen and telogen. Hierarchical clustering analysis clearly distinguished the differentially expressed genes of each stage. GO analysis indicated that these altered genes at proangen were predominantly involved in collagen fibril organization, integrin-mediated signaling pathway, cell-matrix adhesion, cell adhesion, transforming growth factor-β （TGF-β） receptor signaling pathway, regulation of cell growth. Kyoto encyclopedia of genes and genomes （KEGG） analysis showed that the significant pathways involved mainly included focal adhesion and extracellular matrixc （ECM）-receptor interaction. Some important genes involved in these biological processes, such as COL1A1, COL1A2, COL3A1, SPARC, CYR61 and CTGF, were related to tissue remolding and repairing and detected by more than one probe with similar expression trends at different stages of cashmere growth cycle. The different expression of these genes may contribute to understanding the molecular mechanism of cashmere regeneration.     

Effects of Yimu Shenghuasan Preparation on the Cytochrome P450 in Endometrial Cells and Immune Function of Dairy Cows

In order to investigate the mode of action ofYimu Shenghuasan preparation in endometrial cells of dairy cows, the primary cultured endometrial cells in cows were isolated and the inflammatory models were made by lipopolysaccharide （LPS） induction. The inflammatory cells were treated with gradient concentration of herbal medicine preparation, Yimu Shenghuasan for 48 and 72 h. The expression of cytochrome P450 （CYP450） was detected by Western blot. The amounts of IgG and lgA in sera were also detected in the endometritis of dairy cows. The expression level of CYP450 in the endometrial cells of dairy cow was increased gradually, and the amounts of IgG, IgA were increased significantly as compared with those in the control group. The expression level of CYP450 in the inflammatory cells was increased significantly in the treatment of 2 000 μg mL^-1 of Yimu Shenghuasan after 48 h of treatment.     

Physiological traits of rice seedlings in response to inorganic arsenic

The aim of our study was to better understand the different responses of rice seedling to different species of inorganic arsenic As203 （As（Ill）） and Na2HAsO4 （As（V））. Our results indicate that the biomass of rice seedling decreased as arsenic concentration increased, with the decrease being more significant at higher arsenic concentrations. In addition, the analysis of superoxide dimutase （SOD）, peroxidase （POD）, and catalase （CAT） in rice roots and leaves showed that the activity of these three enzymes significantly decreased in rice tissues, especially in rice roots, as arsenic concentration was increased,. Further, the uptake and utilization efficiencies of N, P, and K were found to decrease as arsenic concentration was increased. However, the uptake and utilization efficiencies of P and K were mainly affected by As（IlI）, whereas those of N were mainly affected by As（V）. Inductively coupled plasma-mass spectrometry （ICP-MS） was used to assay arsenic accumulation in rice tissues; the results indicate that the arsenic content in rice tissues was enhanced when arsenic concentration was increased, especially in rice roots after arsenic treatment.     

Fermented Goat Milk and Cow Milk Produced by Different Starters of Lactic Acid Bacteria： Quality Studies

Lactic acid bacteria （LAB） is widely used as culture starters in dairy fermentation. The aim of this study was to investigate the quality of fermented goat milk and cow milk, as well as the viability of LAB in the same products. Fermentations were performed with pasteurized goat milk or cow milk added with skim milk （18% of solids） using three separately different starters; yoghurt starter （a combination of Streptococcus thermophilus FNCC-0040 and Lactobacillus bulgaricus FNCC-0041）, single starter of Lactobacillus acidophilus FNCC-0029 and Lactobacillus casei FNCC-0051. The parameters observed were pH, acidity, nutritional quality including protein, fat and lactose content and product＇s viscosity. Acidity, pH and viability of LAB were also monitored during storage at refrigerated temperature （4 ℃） for 28 days. Results show that the different LAB starters did not affect the pH, acidity, lactose and protein content. Differences on LAB starters affected fat content and viscosity. The highest score of viscosity （30.00 Pa.s ± 7.02 Pa.s） was observed on products fermented by yoghurt starters, followed by products obtained using starter of L. acidophilus （17.7 ±11.4） and L. casei （8.62 ±0.35）. Protein content, acidity, pH and viscosity were not significantly different between products obtained from goat milk and cow milk. Fat content in fermented goat milk was higher （5.03% ±0.62%） than in fermented cow milk （3.52% ±0.37%）, however, lactose content was higher in fermented cow milk （5.16% ±0.40%） than in fermented goat milk （4.53% ±0.35%）. Total LAB concentration in fermented cow milk during storage was 8.03± 0.52 logt0 cfu/mL, while in fermented goat milk was 7.81 loglo cfu/mL ± 0.67 loglo cfu/mL. There was a 10.83% decrease in LAB viability in fermented cow milk and 11.40% in fermented goat milk after 28 days of storage. In conclusion, quality of fermented milk is affected by the starters applied, raw milk source and storage period.     

Variations on conserved signaling pathways in biocontrol and development:G protein and MAPK genes of Trichoderma. atroviride and T. virens

Filamentous fungi employ conserved eukaryotic signaling pathway to detect and respond to environmental signals, including the presence of the host. Genetic experiment in which a particular signaling protein is lost, or its activity enhanced, have defined some of the function of heterotrimeric G proteins and MAP kinases in development and virulence. A hallmark of these studies is that orthologs in different species may have different functions. Antagonistic fungal-fungal interactions form …     

Kaempferol inhibits Pseudorabies virus replication in vitro through regulation of MAPKs and NF-κB signaling pathways

Pseudorabies virus(PRV), in the family Herpesviridae, is a pathogen of Aujeszky's disease, which causes great economic losses to the pig industry. Recent outbreaks of Pseudorabies imply that new control measures are urgently needed. The present study shows that kaempferol is a candidate drug for controlling PRV infection, as it possesses the ability to inhibit PRV replication in a dose-dependent manner in vitro. Kaempferol at a concentration of 52.40 μmol L–1 could decrease PRV-induced cell death by 90%. With an 50% inhibitory concentration(IC50) value of 25.57 μmol L–1, kaempferol was more effective than acyclovir(positive control) which has an IC50 value of 54.97 μmol L–1. A mode of action study indicated that kaempferol inhibited viral penetration and replication stages, decreasing viral loads by 4-and 30-fold, respectively. Addition of kaempferol within 16 h post infection(hpi) could significantly inhibit virus replication, and viral genome copies were decreased by almost 15-fold when kaempferol was added at 2 hpi. Kaempferol regulated the NF-κB and MAPKs signaling pathways involved in PRV infection and changed the levels of the target genes of the MAPKs(ATF-2 and c-Jun) and NF-κB(IL-1α, IL-1β and IL-2) signaling pathways. The findings of the current study suggest that kaempferol could be an alternative measure to control PRV infection.     

The Differentiation-and Proliferation-Inhibitory Effects of Sporamin from Sweet Potato in 3T3-L1 Preadipocytes

The aim of this study was to investigate the effect of different concentrations of sporamin on the differentiation and proliferation of 3T3-LI preadipocytes, providing the theoretical basis for the development of food to treat obesity and diabetes, The isolation and purification of sporamin from sweet potato species 55-2 were performed by ammonium sulphate precipitation in combination with ion-exchange and gel filtration chromatography. With berberine as a positive control, different concentrations ofsporamin （0.000, 0.125, 0.025, 0.250, 0.500, and 1.000 mg·mL^-1 were used to treat 3T3-L1 preadipocytes. Intracellular fat accumulation and the degree of adipogenesis were quantified using Oil Red O staining and colorimetry, Preadipocytes differentiation was measured by 3（4,5-dimethylthiazolyl-2-yl）-2,5-diphenyltetrazolium bromide （MTT） spectrophotometric assay. Two sporamin proteins, which were separated into sporamin A （31 kD） and sporamin B （22 kD）, could be purified by ion-exchange and gel filtration chromatography. After being treated by different concentrations of sporamin, the differentiation of 3T3-L1 preadipocytes was significantly inhibited, compared with the positive control. When the sporamin solution concentration was 0.500 mg mL-1, the accumulation of lipid droplets within the cells was significantly decreased and the optical density （OD） value of the solution from destained Oil Red O reached to 0.35, which was the lowest value （P〈 0.05）. The proliferation of 3T3-L1 preadipocytes was significantly inhibited by treating at higher sporamin concentrations. In addition, the inhibitory effect was more obvious with the prolonged treatment time （P〈 0.05）. The differentiation and proliferation of 3T3-L1 preadipocytes could be inhibited significantly by the addition of higher concentration sporamin. It was, therefore, suggested that the sporamin was potentially effective for weight loss.