miRNA expression analysis of the sheep follicle during the prerecruitment, dominant, and mature stages of development under FSH stimulation.

Follicle stimulating hormone (FSH) has been widely reported to influence ovarian follicular development, and miRNAs play a significant role in mammalian follicular development by regulating their target genes. Therefore, it is of interest to explore the roles of miRNAs in sheep follicular development during FSH stimulation. In the current study, we constructed miRNA expression profiles of small follicles (SFs, prerecruitment stage), medium follicles (MFs, dominance stage), and large follicles (LFs, maturation stage). Three and 50 significant differentially expressed miRNAs (DEMs) were identified in the MF vs. SF and LF vs. SF comparisons, respectively, and none were identified in the LF vs. MF comparison. Oar-miR-10a was significantly downregulated in MFs compared with SFs. In LFs compared with SFs, miR-212-3p, miR-212-5p and miR-202-5p were significantly upregulated, and miR-27a-3p, miR-181a-5p, miR-204-5p, and miR-182-5p were significantly downregulated. Furthermore, we predicted the target genes of significant DEMs and performed functional enrichment analyses of these target genes. Analyses of KEGG pathways and GO terms showed that the putative target genes were significantly enriched in ovarian steroidogenesis, glutathione metabolism, positive regulation of cell differentiation, positive regulation of cell development, and cellular response to oxygen-containing compounds. Analyses of miRNA-gene regulatory networks suggested that miR-181a-5p-CYP11A1, (miR-27a-3p and miR-129-5p)-LDLR, (miR-212-3p and miR-212-5p)-EFNA5, (miR-181a-5p, miR-182-5p, and miR-27a-3p)-INHBA, and miR-182-5p-SOD2 might be involved in follicular development. The present study provides basic data and suggests research directions for further exploration of the roles of miRNAs in sheep follicular development under FSH stimulation.

Transcriptome analysis of sheep follicular development during prerecruitment, dominant, and mature stages after FSH superstimulation.

Sheep is usually a monovular animal; superovulation technology is used to increase the number of offspring per individual and shorten generation intervals. To date, mature FSH superstimulatory treatments have been successfully used in sheep breeding, but much remains unknown about genes, pathways, and biological functions involved in follicular development. Therefore, in this study, we performed transcriptome profiling of small follicles (SFs; 2-2.5 mm), medium follicles (MFs; 3.5-4.5 mm), and large follicles (LFs; > 6 mm) in Mongolian ewes after FSH superstimulation. Furthermore, we identified differentially expressed genes and performed Kyoto Encyclopedia of Genes and Genomes pathway and Gene Ontology enrichment analyses in 3 separate pairwise comparisons. We found that ovarian steroidogenesis was significantly enriched in the SFs versus MFs analysis; the associated genes, cytochrome P450 family 19 (CYP19) and Hydroxy-delta-5-steroid dehydrogenase 3 beta- and steroid delta-isomerase 1 (HSD3B1), were significantly upregulated. Moreover, proline metabolism, glutathione metabolism, and PPAR signaling pathways were significantly enriched in the LFs versus SFs analysis; the associated genes, glutamate-cysteine ligase modifier subunit (GCLM) and cystathionine gamma-lyase (CTH), were significantly upregulated, whereas peroxisome proliferator-activated receptor gamma (PPARγ) was significantly downregulated. In summary, our study provides basic data and possible biological direction to further explore the molecular mechanism of sheep follicular development after FSH superstimulation.

Characterization and structure of glyceraldehyde-3-phosphate dehydrogenase type 1 from Escherichia coli.

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a key enzyme in the glycolytic pathway that catalyzes the conversion of D-glyceraldehyde 3-phosphate to 1,3-diphosphoglycerate. Here, the full-length GAPDH type 1 from Escherichia coli (EcGAPDH1) was cloned and overexpressed, and the protein was purified. Biochemical analyses found that the optimum reaction temperature and pH of EcGAPDH1 were 55°C and 10.0, respectively. The protein has a certain amount of thermostability. Crystals of EcGAPDH1 were obtained using the sitting-drop vapor-diffusion technique and X-ray diffraction data were collected to 1.88 Šresolution. Characterization of the crystals showed that they belonged to space group P41212, with unit-cell parameters a = b = 89.651, c = 341.007 Å, α = ß = γ = 90°. The structure of EcGAPDH1 contains four subunits, each of which includes an N-terminal NAD+-binding domain and a C-terminal catalytic domain. Analysis of the NAD+-bound form showed some differences between the structures of EcGAPDH1 and human GAPDH. As EcGAPDH1 shares 100% identity with GAPDH from Shigella sonnei, its structure may help in finding a drug for the treatment of shigellosis.

[Perceived organizational support and emotional stability modify the effect of depression tendency on burnout in doctors].

Objective: To explore and analyze the relationship between burnout, depression, perceived organizational support and emotional stability, as well as their interaction on burnout, so as to provide reference for improving the situation of burnout on doctors. Methods: By cluster random sampling, a cross-sectional survey of doctors in a municipal hospitals of Zhengzhou in June to August, 2015, Henan Province was conducted. The questionnaires of Maslach Burnout Inventory-General Survey (MBI-GS) , Perceived organizational support, Eysenck Personality Questionnair-Revission Short Scale of China (EPQ-RSC) and Center for Epidemiological Studies Depression Scale (CES-D) were used for questionnaire survey and analysis. Multiple regression analysis was used to explore the interaction of organizational support and emotional stability on depressive tendency and occupational burnout. Results: Among the 389 doctors, 147 were males and 242 were females. Age M (P(25), P(75)) was 33.0 (29.0, 40.0) years, and length of work M (P(25), P(75)) was 7.0 (4.0, 16.0) years. Burnout was positively correlated with depression tendency and emotional stability (r=0.571, 0.453, P<0.01) , while burnout was negatively correlated with perceived organizational support (r=-0.260, P<0.01) . The interaction among depression tendency, perceived organizational support and emotional stability was statistically significant (ß=-0.002, P<0.05) , the independent effect on burnout is 1.0% (ΔR(2)=0.010) , the positive effect of emotional stability was the largest (ß=0.550) , followed by the positive effect of depression tendency (ß=0.494) . When the tendency of depression increased, the burnout of doctors with high perceived organizational support and emotional stability was higher than those with high perceived organizational support and emotional instability and those with low perceived organizational support (P<0.01) . Conclusion: Emotional stability is the main factor affecting the effect of depression tendency on doctors' burnout. Doctors with stable emotion and high perceived organizational support showed obvious changes in burnout when depression tendency increased, and showed stronger burnout.

A general experimental system for the development of acoustic logging tools.

Laboratory testing is a pre-requisite for the practical application of new methods and techniques, and it is crucial in the research and development of acoustic well-logging tools. Various tools have been developed based on different acoustic logging theories and methods. Thus, these tools are equipped with different acoustic sonde structures. To meet the test requirements of different tools in a laboratory environment, we designed a general experimental system that includes hardware platform, software platform, and model wells according to the common structure of actual logging tools. Similar to the internal electrical structure of downhole tools, the hardware platform consists of several main parts, such as power supply, control and telemetering, acoustic emission, and data acquisition. The functions of this hardware platform include controlling the working sequence of the experiment, exciting the transmitter sonde, and collecting the acoustic signals received by the receiver sonde. The software platform installed in the host computer provides a human-computer interface for the experimental system to complete the data transmission between the host computer and the hardware platform, store measured data, and process the data in real time. The model wells approximate the actual engineering environment and stratum condition for system testing. A series of practical laboratory experiments is conducted in the model wells by using this experimental system. The process proves that the hardware and software of the experimental system can work in coordination, and the experimental system meets the basic testing requirements of conventional acoustic logging tools.

[Expression of type 1 and type 2 cytokines from serum of coal miners and the evaluation in surveillance of coal workers' pneumoconiosis at earlier stage].

Objectives: To explore the expression regulation of type 1 and type 2 (Th1 and Th2) cytokines from serum of coal miners and the evaluation in surveillance of coal workers' pneumoconiosis, 630 coal miners were studied. Methods: A total of 90 male patients diagnosed as coal workers' pneumoconiosis (CWP) in a institute for occupational health and 19 male workers newly diagnosed as CWP patients was chosen as CWP group with simple random sampling method from a coal mine group from January 2013 to December in 2015. 180 male coal miners with abnormal but not diagnosed as CWP were selected as CWP suspected group with simple random sampling methods, meanwhile 180 male coal miners with normal chest X-ray photograph was as dust-exposed group by 1∶1 matched as age. And 161 healthy males accepted pre-employed examination were selected as control group, CWP suspected group, dust-exposed group and control group called as non-CWP group. According to screening test and diagnosis test, the basic information and occupational history of all subjects were collected, and cytokines including IL-1ß, IL-8, IFN-γ, IL-6 and IL-10 of serum were detected. Receiver operator characteristic (ROC) curve was used to determine the optimal cutoff value of each cytokine. Area under curve (AUC), the validity and reliability were calculated and judged. Results: The average age of control group, dust-exposed group, CWP suspected group and CWP group were (27.4±5.0) , (43.4±10.7) , (48.2±6.2) , (64.7±7.0) years old, respectively. The median level of IL-1ß, IL-8, IFN-γ and IL-6 in cases group (1 638.30, 2 099.49, 815.18,140.32 pg/ml) were higher than that of non-cases group (1 445.57, 1 402.26, 736.38, 95.73 pg/ml) (P<0.05) . The level of IL-8 (1 503.99 pg/ml) in CWP suspected group was higher than that of control group (1 295.67 pg/ml) and dust-exposed group (1 376.94 pg/ml) , but the level of IL-10 (654.08 pg/ml) was lower than that of control group (596.64 pg/ml) . The ratio of IFN-γ/IL-6 ranged from 5 to 8, and the ratio in CWP group (5.87) was lower than that of non-CWP group (7.61) . The IL-6 and IL-8 among the subjects of dust-exposed group in terms of the age distribution of among had reached statistical significance. According to ROC, the cutoff value of IL-1ß, IL-6, IL-8, IL-10 and INF-γ reached 1 582.65, 116.53, 1 791.54, 581.08 and 792.69 pg/ml, respectively. The AUC was 0.668, 0.895, 0.859, 0.716 and 0.637, respectively. It was found that IL-6 and IL-8 could be used as biomarkers in detecting CWP, the sensitivity and specificity was 82.6% and 84.6%, 78.0% and 84.8%, respectively; Youden's index was 0.674 and 0.628 and the consistency rate was 84.3% and 83.7%, while Kappa value was 0.55 and 0.52. Conclusion: There was Type 1 and type 2 cytokine dysregulation in CWP patients. IL-6 and IL-8 can be used as effective biomarkers to forecast lung injury before X-ray changes.

[Effects of Personality and Psychological Acceptance on Medical Workers' Occupational Stress].

Objective: To assess psychological acceptance and occupational stress of medical staff, analyze the relationship among personality, psychological acceptance and occupational stress and discuss the direct or indirect effects of personality to occupational stress. Methods: Eysenck Personality Questionnaire (EPQ-RSC) , Acceptance and Action Questionnaire-II (AAQ-Ⅱ) and Revised Occupational Stress Inventory (OSI-R) were administered to 749 medical staff. Results: The level of occupational stress of medical staff was high, the score of PSY was 26.8±7.13 and the score of PHS was 24.3±6.50. Personality and psychological acceptance can predict occupational stress. Psychological acceptance was a protective factor of occupational stress. Medical staff with personality of introversion, neuroticism and psychoticism suffered higher occupational stress. Personality have both direct and indirect effects on occupational stress. Neuroticism have the strongest effect on occupational stress with effect size of 0.496 (psychological stress) and 0.431 (physical strain) . Conclusion: Medical staff have heavier occupational stress. There is a significant correlation between personality and occupational stress. Measures depending on personality should be taken to deal with this situation.

Influence of castration-induced sex hormone deficiency on serum lipid levels and the genes expression in male pigs.

It has been known that the changes in gonadal steroids are closely associated with adipose tissue metabolism. Domestic pigs have been a well-recognized experimental animal in biomedical research because of their similarity to humans in body size and other physiological/anatomical features. The aims of this study were to investigate the influence of castration-induced sex hormone deficiency on serum lipid levels and the genes expression of key enzymes associated with lipogenic and lipolytic processes in male pigs. The experimental animals consisted of 2 groups slaughtered on 147th and 210th day, respectively. In each of the group, 7 full-sib pairs of castrated and intact male hybrids from Yorkshire dams sired by Landrace were contained. The results showed that castration of male pigs led to increased total cholesterol, triglyceride, high density lipoprotein, low density lipoprotein, and leptin levels in serum (p<0.05). No differences in levels of the free fatty acid, insulin, and glucose were observed between boars and barrows (p>0.05). Castration caused upregulation of fatty acid synthase and acetyl-CoA carboxylase alpha genes expression at both 147 and 210 days of age (p<0.05). No differences in expression of hormone sensitive lipase and adipose tissue triglyceride lipase genes were observed between boars and barrows at either 147 or 210 days of age (p>0.05). It is speculated that higher body fat deposition in castrated male pigs might have resulted mainly from increased transcription of the lipogenic genes, but not from decreased transcription of the lipolytic genes.

Alkaloids from Cynanchum komarovii with inhibitory activity against the tobacco mosaic virus.

A pyrroloisoquinoline alkaloid, 2,3-dimethoxy-6-(3-oxo-butyl)-7,9,10,11,11a,12-hexahydrobenzo[f]pyrrolo[1,2-b]isoquinoline (1), whose structure was determined by spectroscopic methods, was isolated from the aerial parts of Cynanchum komarovii, together with two known alkaloids, 7-demethoxytylophorine (2) and 7-demethoxytylophorine N-oxide (3). Alkaloids 2 and 3 had antiviral activities against tobacco mosaic virus.

[Detection of Cre recombinase activity in Mx-Cre transgenic mice induced by INF in vitro].

The Cre recombinase and its activity in C57-TgN(Mx-Cre) transgenic mice is studied by polymerase chain reaction (PCR), Western blot, immunohistochemistry, immunogold electron microscopy and Southern blot. C57-TgN(Mx-Cre) transgenic mice harbouring cre gene in genomic DNA is demonstrated by PCR, and these mice which are induced by INF-alpha 1b could express Cre recombinase, which is confirmed by Western blot. With immunohistochemistry, we find that the Cre recombinase expresses in hepatocyte cytoplasm and nuclear of C57-TgN(Mx-Cre) transgenic mice. Cre recombinase expressed in hepatocyte cytoplasm and nuclear is further confirmed by immunogold electon microscopy. And it is supported that the Cre recombinase which is created from C57-TgN(Mx-Cre) transgenic mice induced by INF-alpha 1b can direct DNA recombination reaction in vitro. All evidence leads us supporting the view that the Cre recombinase expressed in C57-TgN(Mx-Cre) transgenic mice has activity. Thus we find a method to detect the activity of Cre recombinase in vitro.

[Expression of Trial in mammalian cell and its apoptosis activation in culture cells].

To develop a system for expression of Trail in mammalian cell. PCR and sequencing were used to clone and to confirm Trail gene. Then, we constructed the vector for expression of Trail in mammalian cells. HeLa cells, one of sensitive cell lines to Trail, were transfected with the vector DNA. After 48 h, flow cytometry was used to screen HeLa transfected by Trail. 19% of total cells happened to apoptosis. This study conforms that the production of Trail expressing in mammal cell can induce apoptosis and exhibit high bio-active. The results indicate the potential application of Trail to tumor immunotherapy.

[Generation of Mx-cre transgenic mice].

To generate the transgenic mice expressing cyclization recombination enzyme, the recombinant gene, in which the coding region of cre gene is derived by the promoter of mouse Mx gene, was microinjected into pronuclei of fertilized mouse eggs. Founders of transgenic mice harbouring the recombinant gene were screened by polymerase chain reaction (PCR) at genomic DNA level and confirmed by Southern blot. One line of Mx-cre transgenic mice was obtained. Then, the Mx-cre transgenic mouse line was cultured and propagated.

[The presentation of neo gene in chimeric mice obtained from ES cells].

To explore the potentiality of the mouse ES cell line which is employed in our laboratory were transfected with neo gene and selected with G418. Some ES cell clones expressing neo gene were obtained. After ES clones were picked up and expanded, the ES cells were mircoinjected into blastocysts and implanted miceuterus. 60 blastocysts were implanted into 5 mice. Finally, 3 chimeric mice with neo gene were born. PCR was carried out to analyze the presentation of neo gene in tissues of chimeric mice. The results showed that many tissues harbored the neo gene, including skin, liver and blood et al.

Bias correction for segregation ratio estimation in human genetics.

Segregation ratio estimation has long been important in human genetics. A simple truncated binomial model is considered that assumes complete ascertainment and a deterministic genotype-phenotype relationship. A simple but intuitively appealing estimator of the segregation ratio, previously proposed, is shown to have a negative bias. It is also shown that the bias of this estimator can be largely reduced via a randomization device, resulting in a new estimator that has the same large-sample behavior but with a negligible bias (decaying at a geometric rate). Numerical results are given to show the small-sample performance of this new estimator. An extension to incomplete ascertainment is also considered.

[Expression of 3'-truncated preS/S gene in transgenic mice].

The 3'-truncated preS/S region from HBV genome, encoding a transcriptional transactivator, was cloned and a recombination expression vector for 3'-truncated HBV preS/S sequences under the control of the CMV promoter was constructed by recombination DNA techniques. Then, the expression vector DNA was microinjected into pronuclei of fertilized mouse oocytes. Founders of transgenic mice harbouring the recombination gene which can be expressed were screened by polymerase chain reaction (PCR) at genomic DNA level and confirmed by ELISA andlysis at protein level. Two of 15 mice in one series of microinjections showed the expression of 3'-truncated preS/S gene from HBV genome. The expression vector of 3'-truncated preS/S gene might be helpful for further studies of relationship between the expression production of 3'-truncated preS/S sequence and HBV-associated oncogenesis in vitro. The transgenic mice with expressing 3'-truncated preS/S gene will provide new insight into its roles in the development of human hepatocelluar carcinoma (HCC) in vivo.