Differentially expressed microRNAs and affected signaling pathways in placentae of transgenic cloned cattle.

Placental deficiencies are related to the developmental abnormalities of transgenic cattle produced by somatic cell nuclear transfer, but the concrete molecular mechanism is not very clear. Studies have shown that placental development can be regulated by microRNAs (miRNAs) in normal pregnancy. Thus, this study screened differentially expressed miRNAs by the next-generation sequencing technology to reveal the relationship between miRNAs expression and aberrant development of placentae produced by the transgenic-clone technology. Expressions of miRNAs and mRNAs in different placentae were compared, the placentae derived from one natural pregnancy counterpart (PNC), one natural pregnancy of a cloned offspring as a mother (PCM), and two transgenic (human beta-defensin-3) cloned pregnancy: one offspring was alive after birth (POL) and the other offspring was dead in 2 days after birth (POD). Further, signaling pathway analysis was conducted. The results indicated that 694 miRNAs were differentially expressed in four placental samples, such as miR-210, miR-155, miR-21, miR-128, miR-183, and miR-145. Signaling pathway analysis revealed that compared with PNC, significantly upregulated pathways in POL, POD, and PCM mainly included focal adhesion, extracellular matrix-receptor interaction, pathways in cancer, regulation of actin cytoskeleton, endosytosis, and adherens junction, and significantly downregulated pathways mainly included malaria, nucleotide binding oligomerization domain-like receptor signaling, cytokine-cytokine receptor interaction, Jak-STAT signaling pathway. In conclusion, this study confirmed alterations of the expression profile of miRNAs and signaling pathways in placentae from transgenic (hBD-3) cloned cattle (PTCC), which could lead to the morphologic and histologic deficiencies of PTCC. This information would be useful for the relative research in future.

Dual effects of molybdenum on mouse oocyte quality and ovarian oxidative stress.

A sub-acute toxicity test was performed to investigate the effects of molybdenum (Mo) on ovarian function. ICR adult female mice were exposed to Mo by free access to distilled water containing the Mo at 5, 10, 20, and 40 mg/L for 14 days. Compared to the control group, M II oocyte morphology, ovary index, and ovulation improved within the 5 mg/L Mo group, but were negatively affected by Mo at 40 mg/L. Morphologically abnormal ovarian mitochondria were observed at ≥ 20 mg/L. These alterations accompanied the changes in superoxide dismutase (SOD), glutathione peroxidise (GPx), and malondialdehyde (MDA) levels in ovaries. In conclusion, Mo affects oocyte quality possibly through regulating ovarian oxidative stress in a dose-dependent manner. It appears that Mo may improve ovarian function at a suitable concentration, which might be a candidate for the treatment of female infertility.

Effects of molybdenum on sperm quality and testis oxidative stress.

In order to investigate the effects of molybdenum (Mo) on sperm parameters and testicular oxidative stress, the ICR strain of adult mice were exposed to different doses of molybdenum for a sub-acute toxicity test. Compared to the control, our results showed that the sperm parameters, including the epididymis index, sperm motility, sperm count, and morphology, increased by a moderate dose of Mo (25 mg/L), but were negatively affected at high doses (≥ 100 mg/L). In addition, the changes of sperm parameters were accompanied with changes of the superoxide dismutase (SOD) activities, the glutathione peroxidase (GPx) activities, and the malondialdehyde (MDA) levels in testes. In conclusion, Mo affects the sperm quality through regulating the testicular oxidative stress in a complex manner.

The effect of molybdenum on the in vitro development of mouse preimplantation embryos.

The object of this study was to investigate the effect of molybdenum on the development of mouse preimplantation embryos cultured in vitro. Zygotes were flushed from one outbred mouse strain (Kunming), and then were cultured in potassium simplex optimized medium (KSOM) containing 0, 5, 10, 20, 40, 80, 120, and 160 µg/ml of molybdenum for 5 days until the mid-blastocyst stage. The addition of ≤ 20 µg/ml molybdenum did not affect the blastocyst and birth rates. Molybdenum at doses of 40 µg/ml and higher significantly decreased the cleavage, blastocyst and birth rates, the average cell number, and significantly increased the proportion of degenerative blastocysts. At 120 µg/ml molybdenum inhibited the blastocysts development to birth. At 160 µg/ml molybdenum caused overall developmental arrest (up to 16-cells) of embryos and their massive degeneration. In conclusion, molybdenum negatively affected the development of embryos in a dose-dependent manner. With lower doses (≤ 20 µg/ml), mouse embryos were not apparently damaged. With very high doses (≥ 40 µg/ml), embryo quality significantly decreased. This assessment of the effect of molybdenum on the preimplantation embryo is an initial survey of toxicological risk.

[Study of cuttings identification using laser-induced breakdown spectroscopy].

Cutting identification is one of the most important links in the course of cutting logging which is very significant in the process of oil drilling. In the present paper, LIBS was used for identification of four kinds of cutting samples coming from logging field, and then multivariate analysis was used in data processing. The whole spectra model and the feature model were built for cuttings identification using PLS-DA method. The accuracy of the whole spectra model was 88.3%, a little more than the feature model with an accuracy of 86.7%. While in the aspect of data size, the variables were decreased from 24,041 to 27 by feature extraction, which increased the efficiency of data processing observably. The obtained results demonstrate that LIBS combined with chemometrics method could be developed as a rapid and valid approach to cutting identification and has great potential to be used in logging field.

Blastocysts cloned from the Putian Black pig ear tissues frozen without cryoprotectant at -80 and -196 degrees Celsius for 3 yrs.

The Putian Black pig, as one of elite cultivars of endemic species in China, has been on the verge of extinction and urgently needs protection. Somatic cell nuclear transfer (SCNT) and noncryoprotected frozen tissue technology have successfully resurrected several mammalian species. Therefore, this study explored the primary feasibility of conserving this breed using a combination of both technologies. Skin tissues obtained from the ears of adult Putian Black boars were frozen without cryoprotectant at -20, -80, or -196 °C and stored for 3 yrs. Primary cell culture, passage and subculture were performed on frozen samples after being rapidly thawed at 39 °C and on fresh pig ear tissues (control). Cloned embryos were reconstructed using fibroblasts (from frozen and fresh tissues) with enucleated oocytes. Live cell lines were obtained from tissues frozen at -80 and at -196 °C and appeared to have normal proliferative activity after passage; furthermore, they directed cloned embryos to develop to the blastocyst stage after nuclear transfer. We concluded that the population of Putian Black pig might be increased in the future by transferring cloned blastocysts into synchronized recipient pigs.

[Affecting factors on capture efficacy of sex pheromone traps for Cydia pomonella L].

Codling moth Cydia pomonella L. (Lepidoptera, Olethreutidae) is the most serious pest of orchards, and one of the most important quarantine pests in China. This paper studied the effects of the color, shape, placement location, lure color, and lure number of sex pheromone traps on the capture efficacy for C. pomonella L. male moth. It was found that the capture efficacy of white and green traps was two times higher than that of blue trap (P < 0.05), and water bottle and triangle traps had no significant difference in their trapping effect. The traps hung in the middle of crown gave two times higher catches than those hung in the upper portion of crown (P < 0.05). Lure color and number had no significant effect on the capture efficacy. The present results could be used in better monitoring C. pomonella by using its sex pheromones.