The development of trade and economic cooperation between China and Belarus and the main challenges it faces

The article describes the stages of development of economic and trade cooperation between China and Belarus, analyzes the reasons that countries face in the process of cooperation and suggests ways to solve them

Numbers of Publications Related to Laboratory Animals

Laboratory animals are widely utilized in biomedical research, so a search of scientific publications can  give us useful information on the use of animals. We retrieved the PubMed biomedicine database and  searched for publications related to laboratory animals from 1966 to 2005. We found that rats and mice  constitute the vast majority of species used in biomedical research; C57BL and BABL/c inbred mice, and  Sprague Dawley and Wistar outbred rats are the most common strains. Recently, the numbers of publications  relating to traditionally used animals such as rats, guinea pigs, dogs, cats, and sheep decreased slightly,  whereas the numbers relating to mice, fish, Drosophila and Caenorhabditis elegans increased from 1995  to 2005, with annual mean growth rates of 4.5%, 8.22%, 1.95%, and 10.3%, respectively. Publications  involving transgenic mice increased dramatically from the mid-1980s. This survey provides significant  clues for predicting the future direction of biomedical research.

Factors Influencing the Number of Eggs Recovered from Rabbits Superovulated with FSH or PMSG: Analysis of Five Years of Data from 509 Rabbits

To determine the best conditions for superovulation in rabbits, we analyzed the influence of age, season and hormone treatment on the numbers of eggs collected over five years from 509 rabbits aged 4–10 months using follicular stimulating hormone (FSH) or pregnant mare serum gonadotropin (PMSG) hormone stimulation. The number of eggs recovered was significantly higher in younger rabbits in both treated groups (P < 0.01–0.05). The number of eggs collected from rabbits treated with FSH were significantly higher than from rabbits treated with PMSG at all ages (P < 0.01). Seasonal differences were not observed in either hormone treatment group as they were maintained under constant temperature, humidity and light cycle through the year. Thus, younger rabbits are more sensitive to hormonal superovulation treatment with both FSH and PMSG, and FSH offers a better regimen for egg collection

A novel CpG ODN compound adjuvant enhances immune response to spike subunit vaccines of porcine epidemic diarrhea virus

CpG oligodeoxynucleotides (CpG ODNs) boost the humoral and cellular immune responses to antigens through interaction with Toll-like receptor 9 (TLR9). These CpG ODNs have been extensively utilized in human vaccines. In our study, we evaluated five B-type CpG ODNs that have stimulatory effects on pigs by measuring the proliferation of porcine peripheral blood mononuclear cells (PBMCs) and assessing interferon gamma (IFN-γ) secretion. Furthermore, this study examined the immunoenhancing effects of the MF59 and CpG ODNs compound adjuvant in mouse and piglet models of porcine epidemic diarrhea virus (PEDV) subunit vaccine administration. The in vitro screening revealed that the CpG ODN named CpG5 significantly stimulated the proliferation of porcine PBMCs and elevated IFN-γ secretion levels. In the mouse vaccination model, CpG5 compound adjuvant significantly bolstered the humoral and cellular immune responses to the PEDV subunit vaccines, leading to Th1 immune responses characterized by increased IFN-γ and IgG2a levels. In piglets, the neutralizing antibody titer was significantly enhanced with CpG5 compound adjuvant, alongside a considerable increase in CD8+ T lymphocytes proportion. The combination of MF59 adjuvant and CpG5 exhibits a synergistic effect, resulting in an earlier, more intense, and long-lasting immune response in subunit vaccines for PEDV. This combination holds significant promise as a robust candidate for the development of vaccine adjuvant

Effect of the primary cooling rate on the motility and fertility of frozen-thawed rabbit spermatozoa

[EN] In the present study, we examined the effect of primary cooling rates on the motility and fertility of frozen-thawed rabbit spermatozoa. Rabbit semen diluted with an egg-yolk acetamide extender was cooled from room temperature to 5°C at four different rates (-0.1, -0.2, -0.4, -0.8°C/min) as a primary cooling step, then semen was frozen in liquid nitrogen vapour. After thawing, sperm cooled at -0.1°C/min showed the highest motility (40.7 ± 7.3%); there were no significant differences between the motilities of the -0.1, -0.2, and -0.4°C/min groups. The motility of frozen-thawed sperm cooled at -0.8°C/min (29.2 ± 6.8%) was significantly lower than that of sperm cooled at -0.1 and -0.2°C/min. The viability (-0.1°C/min, 38.1 ± 4.0%; -0.8°C/min, 24.3 ± 7.3%) of frozen-thawed sperm was closely related to its motility (-0.1°C/min, 36.7 ± 7.2%; -0.8°C/min, 22.3 ± 4.7%). Quality of post-thaw motile sperm cooled at different rates was estimated by comparing the fertilisation ability of the -0.1 and -0.8°C/min groups following artificial insemination. There were no significant differences in pregnancy rates and mean litter sizes. These data suggest that cooling rabbit semen at rates ranging from -0.1 to -0.8°C/min affects the viability but not the fertilisation capacity of motile spermatozoa after thawing.The authors thank Ms. T. Shimazaki, Ms. R. Tsuneyoshi, and Ms. R. Eriguchi for their technical assistance. This study was partly supported by Grants-in-Aid for Scientific Research from the Japan Society for the Promotion of Science (No. 22500386).Maeda, T.; Liu, E.; Nishijima, K.; Tanaka, M.; Yamaguchi, S.; Morimoto, M.; Watanabe, T.... (2012). Effect of the primary cooling rate on the motility and fertility of frozen-thawed rabbit spermatozoa. World Rabbit Science. 20(2):65-70. doi:10.4995/wrs.2012.1080SWORD657020

MiR-3188 Inhibits Non-small Cell Lung Cancer Cell Proliferation Through FOXO1-Mediated mTOR-p-PI3K/AKT-c-JUN Signaling Pathway

This study investigated the role of miR-3188 on the proliferation of non-small cell lung cancer cells and its relationship to FOXO1-modulated feedback loop. Two non-small cell lung cancer (NSCLC) cell lines A549 and H1299 were used. RNA silencing was achieved by lentiviral transfection. Cell proliferation was assessed by immunohistochemical staining of Ki67 and PCNA, Edu incorporation, and colony formation assay. Western blotting was used to examine expression of FOXO1, mTOR, p-mTOR, CCND1, p21, c-JUN, AKT, pAKT, PI3K, p-PI3K, and p27 proteins. It was found that miR-3188 reduced cell proliferation in NSCLC cells. Molecular analyses indicated that the effect of mammalian target of rapamycin (mTOR) was directly mediated by miR-3188, leading to p-PI3K/p-AKT/c-JUN inactivation. The inhibition of this signaling pathway further caused cell-cycle suppression. Moreover, FOXO1 was found to be involved in regulating the interaction of miR-3188 and mTOR through p-PI3K/p-AKT/c-JUN signaling pathway. Taken together, our study demonstrated that miR-3188 interacts with mTOR and FOXO1 to inhibit NSCLC cell proliferation through a mTOR-p-PI3K/AKT-c-JUN signaling pathway. Therefore, miR-3188 might be a potential target for the treatment of NSCLC

Temporal and Quantitative Analysis of Atherosclerotic Lesions in Diet-Induced Hypercholesterolemic Rabbits

The diet-induced atherosclerotic rabbit is an ideal model for atherosclerosis study, but temporal changes in atherosclerotic development in hypercholesterolemic rabbits are poorly understood. Japanese white rabbits were fed a high-cholesterol diet to induce sustained hypercholesterolemia, and each group of 10–12 animals was then sacrificed at 6, 12, 16, or 28 weeks. The rabbit aortas were harvested, and the sizes of the gross and intima atherosclerotic lesions were quantified. The cellular component of macrophages (Mφs) and smooth muscle cells (SMCs) in aortic intimal lesions was also quantified by immunohistochemical staining, and the correlation between plasma cholesterol levels and the progress of atherosclerotic lesions was studied. The ultrastructure of the atherosclerotic lesions was observed by transmission electron microscopy (TEM). Widely variable atherosclerotic plaques were found from 6 weeks to 28 weeks, and the lesional progress was closely correlated with cholesterol exposure. Interestingly, a relatively reduced accumulation of Mφ, an increased numbers of SMCs, and a damaged endothelial layer were presented in advanced lesions. Moreover, SMCs were closely correlated with cholesterol exposure and lesional progress for the whole period. Cholesterol exposure directly determines atherosclerotic progress in a rabbit model, and the changes in the cellular component of advanced lesions may affect plaque stability in an atherosclerotic rabbit model

Construction of a camelid VHH yeast two-hybrid library and the selection of VHH against haemagglutinin-neuraminidase protein of the Newcastle disease virus

Humoral immune response after immunization. Sera from IIama was collected, two-fold diluted and tested by HI using LaSota as antigen. Figure S1 Amplification of VHH through a nested PCR. (A) First round PCR to separate VH from VHH. The upper 900 bp bands represent the VH-CH1-Hinge-CH2 of conventional Abs (lane 1–8). The lower 600 bp bands represent the VHH-Hinge-CH2 of HCAbs (lane 1–8). (B) VHH amplified through nested PCR using 600 bp fragment recovered from first round PCR as template (lane 1–4). M in A and B was the DL2000 DNA marker. C in A and B represent the negative control. Figure S2 PCR identification of inserted VHH. 47 clones were randomly picked to determine the library functional diversity by PCR using universal primers T7 and 3’AD (Table 1). Meanwhile, Sterile water was used as negative controls. 45 clones have amplified the 500 bp VHH fragments (lane 1–47), while negative templates control haven’t amplified any bands (lane C). M indicated the DL2000 DNA marker. Figure S3 Detection of library capacity and library titer. (A) 10-3 dilution plating of the transformed cells calculated a library capacity of 1.25 × 107 independent clones. (B) 10-5 dilution plating of the cultured library indicated a library titer of 3.45 × 108 cfu/mL. Figure S4 Deduced amino acid aligment of 10 random picked VHH. Deduced amino acid sequences were analyzed according to the Kabat numbering. Differences in the sequences are pinked, and the dash represent the missing sequences. Two hallmark Cys residues are labeled by the thick-line boxes. The four conservative hallmark residues of VHH in FR2 are labeled by the dotted line boxes. Figure S5 pGBKT7-HN bait plasmid construction. (A) PCR was carried out to amplify a truncate HN gene (without transmembrane region) from La Sota strain. M, 5000 DNA marker. 1, Truncate HN. C, Negative control. (B) A truncate HN was cloned into pGBKT7 through BamH I and Sal I. M, 5000 DNA marker. 1, Double restriction enzyme digestion of pGBKT7-HN. Figure S6 pHSIE-VHH plasmid construction. (A) 7 positive VHH fragment were amplified from recovered positive clones containing pGADT7-VHH by PCR. M, 5000 DNA marker. 1–7, VHH 1–7. C, Negative control. (B) Double restriction enzyme digestion of pHSIE-VHHs. M, 5000 DNA marker. 1–7, pHSIE-VHH 1–7. Figure S7 Western blot analysis of bait protein expression. 2 mL of Y2HGold(pGBKT7-HN) culture liquid was extracted using yeast protein extraction reagent (Takara). c-Myc tag monoclonal antibody (1:4000 dilution) was used as first antibody and HRP-labeled goat anti-mouse antibody (1:5000) was used as second antibody. The immunoreactive was visualized with cECL Plus Western blotting detection reagent (CWBIO). (DOC 1129 kb

Effect of Pulsed Electric Field Treatment on N-Hydroxyacetylneuraminic Acid Content and Eating Quality of Pork

This study investigated how to reduce the content of N-hydroxyacetylneuraminic acid (Neu5Gc) in pork. By using Neu5Gc standard solution, porcine submandibular gland mucin (PSM) solution, meat pieces and minced meat as experimental subjects, and high performance liquid chromatography with fluorescence detection (HPLC-FLD) for quantitative analysis, the effect of pulsed electric field (PEF) treatment (with 50 cycles of electric shock at 1–4 kV/cm) on the content of total and free Neu5Gc and eating quality attributes (pH, color difference and texture quality) in pork was explored. The results showed that the content of free Neu5Gc in Neu5Gc standard solution showed a decreasing trend as PEF intensity increased, and significantly decreased at PEF intensity of 3 and 4 kV/cm (P 0.05). Therefore, PEF can effectively reduce the content of Neu5Gc in pork and the chewiness and elasticity of pork pieces, but it also shows an adverse effect on the pH and color of pork