The tigecycline resistance gene tetX has an expensive fitness cost based on increased outer membrane permeability and metabolic burden in Escherichia coli.

Livestock-derived tetX-positive Escherichia coli with tigecycline resistance poses a serious risk to public health. Fitness costs, antibiotic residues, and other tetracycline resistance genes (TRGs) are fundamental in determining the spread of tetX in the environment, but there is a lack of relevant studies. The results of this study showed that both tetO and tetX resulted in reduction in growth and an increased in the metabolic burden of E. coli, but the presence of doxycycline reversed this phenomenon. Moreover, the protection of E. coli growth and metabolism by tetO was superior to that of tetX in the presence of doxycycline, resulting in a much lower competitiveness of tetX-carrying E. coli than tetO-carrying E. coli. The results of RNA-seq showed that the increase in outer membrane proteins (ompC, ompF and ompT) of tetX-carrying E. coli resulted in increased membrane permeability and biofilm formation, which is an important reason for fitness costs. Overall, the increased membrane permeability and metabolic burden of E. coli is the mechanistic basis for the high fitness cost of tetX, and the spread of tetO may limit the spread of tetX. This study provides new insights into the rational use of tetracycline antibiotics to control the spread of tetX.

CMKLR1 deficiency maintains ovarian steroid production in mice treated chronically with dihydrotestosterone.

Elevated serum chemerin levels correlate with increased severity of polycystic ovary syndrome (PCOS). However, the role of CMKLR1 signaling in ovarian biology under conditions of excess DHT remains unclear. In this study we compared the effects of continuous 90-day high dose DHT exposure (83.3 □g/day) on wild type and CMKLR1-deficient mice. DHT induced PCOS-like clinical signs in wild type mice as well as significant changes in the expression of hormone receptors, steroid synthesis enzymes, and BMPs and their receptors. In contrast, CMKLR1-deficient mice significantly attenuated DHT-induced clinical signs of PCOS and alterations in ovarian gene expression. To determine whether the BMP4 signaling pathway was involved in the pathogenic effects of CMKLR1 signaling in DHT-induced ovarian steroidogenesis, antral follicles were isolated from wild type and CMKLR1 knockout (KO) mice and treated in vitro with combinations of hCG, DHT, and BMP4 inhibitors. BMP4 inhibition attenuated the induction effects of hCG and DHT on estrogen and progesterone secretion in CMKLR1 KO mice, but not in WT mice, implicating the BMP4 signaling pathway in the CMKLR1-dependent response to DHT. In conclusion, CMKLR1 gene deletion attenuates the effects of chronic DHT treatment on ovarian function in experimental PCOS, likely via BMP4 signaling.

An all permanent magnet electron cyclotron resonance ion source for heavy ion therapy.

A high charge state all permanent Electron Cyclotron Resonance ion source, Lanzhou All Permanent ECR ion source no. 3-LAPECR3, has been successfully built at IMP in 2012, which will serve as the ion injector of the Heavy Ion Medical Machine (HIMM) project. As a commercial device, LAPECR3 features a compact structure, small size, and low cost. According to HIMM scenario more than 100 eµA of C(5+) ion beam should be extracted from the ion source, and the beam emittance better than 75 π*mm*mrad. In recent commissioning, about 120 eµA of C(5+) ion beam was got when work gas was CH4 while about 262 eµA of C(5+) ion beam was obtained when work gas was C2H2 gas. The design and construction of the ion source and its low-energy transportation beam line, and the preliminary commissioning results will be presented in detail in this paper.

[Molluscicidal effect of buried niclosamide sustained-release method in plateau mountainous areas].

OBJECTIVE: To evaluate the molluscicidal effect of the buried niclosamide sustained-release method in Oncomelania snail-infested terraced environments of mountainous areas in Yunnan Province. METHODS: A piece of relatively isolated snail-infested terraced environment was selected as experimental plot and randomly divided into four groups. The buried niclosamide sustained-release methods were performed in Group A, B, C with the doses of 6 g/m2, 12 g/m2, 24 g/m2 respectively, and Group D was sprayed with 50% niclosamide ethanolamine salt wettable powder (2 g/m2) as the control. RESULTS: In Group C (with the buried niclosamide sustained-release method of 24 g/m2), the half-year reduction of living snail density was 89.90%, and one-year reduction of living snail density was 96.80%, which were significantly higher than those of Group D (40.35% and 59.11%) (Both P < 0.01) CONCLUSION: The buried niclosamide sustained-release method is effective and suitable for snail control in mountainous terrace areas.

[Measuring the estrus cycle and its effect on superovulation in mice].

This study compares superovulation efficiency during different stages of the estrus cycle in mice by investigating the pudendum appearance, vaginal epithelial cell type, and the follicular development dynamics, in relation to the estrus cycle. We determined the stages of the estrus cycle by observing the pudendum and making vaginal smears of the Kunming mouse, and ascertaining the follicular development by making paraffin sections. Based on the observation, mice in stages of proestrus, oestrus, metoestrus and dioestrus were superovulated respectively, and their follicular developments and embryo collection results were tracked to ascertain the appropriate estrus cycle stage for superovulation. Our data showed that pudendum appearance and vaginal smears could effectively distinguish different stages of the estrus cycle and that ovarian tissue observation showed that follicular development, corpus luteum formation and luteolysis were regular during the estrus cycle. Likewise, during the estrus cycle of the Kunming mouse, pudendum appearance, vaginal epithelial cell type and follicular development were relational to one another. Superovulating mice in different stages of the estrus cycle showed marked differences both in pregnancy rate and average number of embryos collected. Proestrus was significantly better than metoestrus and dioestrus (P<0.05), and better than oestrus, but the difference between them was not significant (P>0.05). We suggest that pudendum appearance and vaginal smears can be used to distinguish estrus cycle stages and that proestrus is the optimal stage of the estrus cycle for superovulation.

[Somatic nuclear transplantation and serial nuclear transplantation of human finger-domain lacking t-PA gene in goat].

In order to research developmental competence of transgenic somatic cell by serial nuclear transplantation, goat cloned embryos were compared with recloned embryos in ability of in vitro development. Fetal fibroblasts including human finger-domain lacking t-PA gene was microinjected into cytoplasm of the MII oocytes. Goat embryos (G0) were cloned by this procedure. A single blastomere from 16 - 64-cell goat cloned embryos (G0) was microinjected into Intracytoplasm of the MII oocytes. Goat embryos (G) were cloned by this procedure. Goat embryos (G2, G3) were recloned by using 16 - 64-cell recloned embryos. The developmental time of donor embryo affected the developmental rate of recloned embryos (G1, G2). The results show: the cleavage rate of cloned embryos (G0) (76.45% +/- 1.17%) was no difference significantly with recloned embryos (G1 G2 G3) (72.18% +/- 1.97%, 76.05% +/- 2.38%, 75.99% +/- 2.84%); the developmental rate of morulae and blastocysts of cloned embryos (47.20% +/- 2.93%, 11.00% +/- 1.42%) were higher than these of recloned embryos(34.99% +/- 2.66%, 28.23% +/- 2.00%, 23.34% +/- 1.99%) (3.87% +/- 0.67%, 2.08% +/- 1.66%, 0); the morulae rate(29.57% +/- 1.53%, 24.43% +/- 1.87%) and blastocysts rate(1.96% +/- 1.31%, 2.01% +/- 1.34%) of recloned embryos (G1 G2) from 16-cell recloned embryos were lower than those(34.32% +/- 1.31%, 29.76% +/- 1.66% and 3.86% +/- 1.03%, 3.48% +/- 0.34% )from 32 - 64-cell recloned embryos (P > 0.05). In conclusion, nuclear transfer embryos should not were recloned mostly; and the embryos recloned by using 32 - 64-cell embryos achieved higher developmental ability compared with using 16-cell embryos.

Nitric oxide affects preimplantation embryonic development in a rotating wall vessel bioreactor simulating microgravity.

Microgravity was simulated with a rotating wall vessel bioreactor (RWVB) in order to study its effect on pre-implantation embryonic development in mice. Three experimental groups were used: stationary control, rotational control and clinostat rotation. Three experiments were performed as follows. The first experiment showed that compared with the other two (control) groups, embryonic development was significantly retarded after 72 h in the clinostat rotation group. The second experiment showed that more nitric oxide (NO) was produced in the culture medium in the clinostat rotation group after 72 h (P<0.05), and the nitric oxide synthase (NOS) activity in this group was significantly higher than in the controls (P<0.01). In the third experiment, we studied apoptosis in the pre-implantation mouse embryos after 72 h in culture and found that Annexin-V staining was negative in the normal (stationary and rotational control) embryos, but the developmentally retarded (clinostat rotation) embryos showed a strong green fluorescence. These results indicate that microgravity induced developmental retardation and cell apoptosis in the mouse embryos. We presume that these effects are related to the higher concentration of NO in the embryos under microgravity, which have cause cytotoxic consequences.