Effect of fixed-time artificial insemination on corpus luteum gene expression at the day 16 and 25 pregnancy of gilt.

Utilization of female livestock can be optimized by application of Fixed-Time Artificial Insemination (FTAI), which plays an important role in large-scale livestock production. However, molecular mechanism of FTAI affecting reproductive performance remains unclear. To investigate the effects of FTAI on corpus luteum in 16 and 25 days of pregnancy gilts, 12 pregnancy gilts were selected from Altrenogest + PMSG + GnRH (APG) group and control group. The number of left and right CL in APG group were significantly higher than control (p < 0.05). Furthermore, result showed the number of differentially expressed genes between 16APG group and 16 C group was obviously larger than that between 25APG and 25 C group (2394 vs. 1476). Up regulated genes in APG were mainly associated with cytokine production and secretion, cell communication and transport (day 16) and angiogenesis, cell cycle and cell-cell signaling (day 25). The expression of differentially expressed genes (RPL10, CYP17A1, DCN, IL18, LDLR and PRLR) in luteal tissue were verified by real-time PCR. In summary, APG program significantly improve reproductive efficiency of gilts through up regulation of cytokine production/secretion, cell communication and transport in Day 16 pregnancy and angiogenesis, cell cycle and cell-cell signaling at Day 25 pregnancy in porcine.

NPC1b as a novel target in controlling the cotton bollworm, Helicoverpa armigera.

BACKGROUND: Insects cannot synthesize sterols and must acquire them from food. The mechanisms underlying how insects uptake dietary sterols are largely unknown except that NPC1b, an integral membrane protein, has been shown to be responsible for dietary cholesterol uptake in Drosophila melanogaster. However, whether NPC1b orthologs in other insect species, particularly the economically important pests, function similarly remains to be determined. RESULTS: In this study, we characterized the function of NPC1b in Helicoverpa armigera, a global pest that causes severe yield losses to many important crops. Limiting dietary cholesterol uptake to insects significantly inhibited food ingestion and weight gain. Compared to the wild-type H. armigera, the CRISPR/Cas9-edited NPC1b mutant larvae were incapable of getting adequate cholesterol and died in their early life stage. Gene expression profile and in situ hybridization analyses indicated that NPC1b was mainly expressed in the midgut where dietary cholesterol was absorbed. Expression of NPC1b was also correlated with the feeding life stages and was especially upregulated during early larval instars. Protein-ligand docking and sequence similarity analyses further demonstrated that NPC1b proteins of lepidopteran insects shared a relatively conserved cholesterol binding region, NPC1b_NTD, which, however, was highly divergent from bees-derived sequences. CONCLUSION: NPC1b was crucial for dietary cholesterol uptake and growth of H. armigera, and therefore could serve as an insecticide target for the development of a novel pest-management approach to control this economically significant insect pest with little off-target effect on bees and sterol-autotrophic animals. © 2020 Society of Chemical Industry.

[Effects of salidroside on the secretion of inflammatory mediators induced by lipopolysaccharide in the co-culture of rat alveolar macrophages and type II alveolar epithelial cells].

The aim of the present study was to investigate the effect of salidroside (Sal) on inflammatory activation induced by lipopolysaccharide (LPS) in the co-culture of rat alveolar macrophages (AM) NR 8383 and type II alveolar epithelial cells (AEC II) RLE-6TN. CCK-8 colorimetric method was used to detect cell proliferation percentage. The enzyme-linked immunosorbent assay (ELISA) was used to determine the content of tumor necrosis factor alpha (TNF-α), macrophage inflammatory protein-2 (MIP-2) and interleukin-10 (IL-10) in the supernatant. Western blot was used to examine the expression levels of phosphorylated AKT (p-AKT) and total AKT protein. The results showed that pretreatment of RLE-6TN cells or co-culture of RLE-6TN and NR 8383 cells with 32 and 128 µg/mL Sal for 1 h, followed by continuous culture for 24 h, significantly increased the cell proliferation (P < 0.05). Compared with control group, 32 and 128 µg/mL Sal pretreatment significantly increased the ratio of p-AKT/AKT in RLE-6TN cells (P < 0.05). Pretreatment of 32 µg/mL Sal not only inhibited the secretion of TNF-α and MIP-2 by NR 8383 cells induced by LPS (P < 0.05), but also enhanced the inhibitory effect of RLE-6TN and NR 8383 cells co-culture on the secretion of TNF-α and MIP-2 by NR 8383 cells induced by LPS (P < 0.05). In addition, 32 µg/mL Sal pretreatment promoted LPS-induced IL-10 secretion by NR 8383 cells (P < 0.05), and enhanced the promoting effect of co-culture of RLE-6TN and NR 8383 cells on the IL-10 secretion by LPS-induced NR 8383 cells (P < 0.05). In conclusion, Sal may directly inhibit LPS-induced inflammatory activation of AM (NR 8383), promote the proliferation of AEC II (RLE-6TN) through PI3K/AKT signaling pathway, and enhance the regulatory effect of AEC II on LPS-induced inflammatory activation of AM.

[Simultaneous determination of daphnetin, daphnoretin, daphneticin in rat plasma by LC-MS/MS and its application in pharmacokinetic study].

To establish HPLC-MS/MS method for simultaneous determination of daphnetin, daphnoretin, and daphneticin in rat plasma after oral and intravenous administration of Daphne giraldii extract, and then use them in the calculation of pharmacokinetic parameters. Six sprague-dawley rats received intragastric administration of D. giraldii extract (daphnetin, daphnoretin and daphneticin were 88.40, 3.24 and 4.28 mg•kg⁻¹, respectively). Their drug plasma concentration was determined by LC-MS/MS with schisandrin as an internal standard to draw plasma concentration-time curve. The pharmacokinetic parameters were calculated by Kinetica 4.4. The results showed that the linear range was 5-1 000 µg•L⁻¹ for daphnetin, daphnoretin and daphneticin, and the method ological test showed conformance to the requirements.The intraday and inter-day variable coefficients (RSD) were both less than 15.0%, indicating that both of legitimate precise and accuracy were consistent with the analysis requirements of biological samples. For daphnetin, the pharmacokinetic parameters Tmax, Cmax, AUC0-t, T1/2 and MRT were 4 h, 858.96 µg•L⁻¹, 10 566.4 µg•L⁻¹â€¢h, 5.19 h and 9.43 h, respectively. For daphnoretin, the pharmacokinetic parameters Tmax, Cmax, AUC0-t, T1/2 and MRT were 2.92 h, 178.00 µg•L⁻¹, 905.89 µg•L⁻¹â€¢h, 3.50 h and 6.95 h, respectively. For daphneticin, the pharmacokinetic parameters Tmax, Cmax, AUC0-t, T1/2 and MRT were 2 h, 36.67 µg•L⁻¹, 355.11 µg•L⁻¹â€¢h, 4.95 h and 8.27 h, respectively. The LC-MS/MS analysis method established in this study was proved to be so accurate and sensitive that it can be applied to the pharmacokinetic study of daphnetin, daphnoretin and daphneticin.

Preparation and physicochemical characterization of a solid dispersion of (3, 5, 6-trimethylpyrazin-2-yl) methyl 3-methoxy-4-[(3, 5, 6-trimethylpyrazin-2-yl) methoxy] benzoate (VA-T) and polyvinylpyrrolidone.

Ischemic brain injury is a major disease which threatens human health and safety. (3, 5, 6-trimethylpyrazin-2-yl) methyl 3-methoxy-4-[(3, 5, 6-trimethylpyrazin-2-yl) methoxy] benzoate (VA-T), a newly discovered lead compound, is effective for the treatment of ischemic brain injury and its sequelae. But the poor solubility of VA-T leads to poor dissolution and limited clinical application. In order to improve the dissolution of VA-T, the pharmaceutical technology of solid dispersions was used in the present study. VA-T/polyvinylpyrrolidone (PVP) solid dispersion was prepared by the solvent method. The dissolution studies were carried out and solid state characterization was evaluated by differential scanning calorimetry (DSC), infrared spectroscopy (IR), x-ray diffraction (XRD) and scanning electron microscopy (SEM). The dissolution rate of VA-T was significantly improved by solid dispersion compared to that of the pure drug and physical mixture. The results of DSC and XRD indicated that the VA-T solid dispersion was amorphous. The IR spectra showed the possible interaction between VA-T and PVP was the formulation of hydrogen bonding. The SEM analysis demonstrated that there was no VA-T crystal observed in the solid dispersions. The ideal drug-to-PVP ratio was 1:5. In conclusion, the solid dispersion technique can be successfully used for the improvement of the dissolution profile of VA-T.

Remission of rheumatoid arthritis and potential determinants: a national multi-center cross-sectional survey.

The aim of this study is to investigate the remission rate of rheumatoid arthritis (RA) in China and identify its potential determinants. A multi-center cross-sectional study was conducted from July 2009 to January 2012. Data were collected by face-to-face interviews of the rheumatology outpatients in 28 tertiary hospitals in China. The remission rates were calculated in 486 RA patients according to different definitions of remission: the Disease Activity Score in 28 joints (DAS28), the Simplified Disease Activity Index (SDAI), the Clinical Disease Activity Index (CDAI), and the American College of Rheumatology/European League Against Rheumatism (ACR/EULAR) Boolean definition. Potential determinants of RA remission were assessed by univariate and multivariate analyses. The remission rates of RA from this multi-center cohort were 8.6% (DAS28), 8.4% (SDAI), 8.2% (CDAI), and 6.8% (Boolean), respectively. Favorable factors associated with remission were: low Health Assessment Questionnaire (HAQ) score, absence of rheumatoid factor (RF) and anti-cyclic citrullinated peptide (anti-CCP), and treatment of methotrexate (MTX) and hydroxychloroquine (HCQ). Younger age was also predictive for the DAS28 and the Boolean remission. Multivariate analyses revealed a low HAQ score, the absence of anti-CCP, and the treatment with HCQ as independent determinants of remission. The clinical remission rate of RA patients was low in China. A low HAQ score, the absence of anti-CCP, and HCQ were significant independent determinants for RA remission.

[Esthetic reconstruction of the anterior teeth area following a combined periodontic-orthodontic treatment in adult periodontal patients].

OBJECTIVE: To evaluate the role of the combined periodontic-orthodontic treatment in the esthetic reconstruc- tion of the anterior teeth area following periodontitis. METHODS: Thirteen adult patients with anterior teeth displacements were treated. The probing pocket depth (PD; 102 teeth, 612 sites), bleeding on probing (102 teeth, 204 sites), papilla index (PI; 128 papillae), and papillary height (PH; 128 papillae) of each patient were assessed at baseline, 3 months after the initial therapy, and the end of the orthodontic treatment. Non-parametric and paired-sample t tests were carried out for the statistical analysis of the data. RESULTS: Three months after initial therapy, the sites with PD ≤ 3 mm accounted for 79.58% (487/612) of the observed teeth, and 88.73% (181/204) of the buccal and lingual sites of the teeth showed negative bleeding on probing. These findings were better than those at baseline [26.31% (161/612) and 22.06% (45/204), respectively] (P < 0.05), but no sig- nificant difference was observed compared with pro-orthodontic treatment (P > 0.05). Prior to orthodontic treatment, the levels of the PI of 8 and 21 papillae were III and II, respectively, among the 128 observed papillae. After the orthodontic treatment, 51 papillae were at level III and 68 papillae were at level II. The PH of the 102 papillae was 2.84 mm ± 0.62 mm after ortho- dontic treatment. This result indicated significant difference compared with that of pre-orthodontic treatment (1.69 mm ± 0.57 mm) (P < 0.05). CONCLUSION: After initial therapy, moderate orthodontic teeth movements may reconstruct the interproximal soft tissue, with esthetic improvement of the papillary level and resolution of the periodontal defects.

[Study on applicability of kinetic model for water extracts from puerariae radix].

To establish an appropriate experimental and data processing method on the basis of the general kinetic model for extraction of traditional Chinese medicines, in order to study the effect of total flavonoids in water extracts from Puerariae Radix on the adaptability of the model, with total flavonoids of Puerariae Radix as the determination indicator. The results showed that the natural logarithm of mass concentration of total flavonoids showed a good linearity with the changes in extraction time and solvent volume. Through calculating and fitting, we successfully established the kinetic model for water extraction of total flavonoids from Puerariae Radix, and verified its accuracy. Its good fitting degree and controllable deviation within the range of industrial production requirements indicated a good adaptability of the model. However, its equation correction factors require further studies.

Systematic analysis of human microRNA divergence based on evolutionary emergence.

MicroRNAs (miRNAs) play important roles in post-transcriptional gene expression control. To gain new insight into human miRNAs, we performed comprehensive sequence-based homology search for known human miRNAs to study the evolutionary distribution of human miRNAs. Furthermore, we carried out a series of studies to compare various features for different lineage-specific human miRNAs. Our results showed that major expansions of human miRNA genes coincide with the advent of vertebrates, mammals and primates. Further system-level analysis revealed significant differences in human miRNAs that arose from different evolutionary time points for a number of characteristics, implicating genetic and functional diversification for different human miRNAs during evolution. Our finds provide more useful knowledge for further exploring origins and evolution of human miRNA genes.

Laparoscopic repair of delayed-onset ureter injury.

Ureter injury is typically a rare complication, albeit with serious sequelae, of a hysterectomy procedure, especially after laparoscopic hysterectomy. Herein we report on a successful laparoscopic reanastomosis of the ureter conducted at the ureterovesical junction.

Single-nucleotide polymorphism-gene intermixed networking reveals co-linkers connected to multiple gene expression phenotypes.

Gene expression profiles and single-nucleotide polymorphism (SNP) profiles are modern data for genetic analysis. It is possible to use the two types of information to analyze the relationships among genes by some genetical genomics approaches. In this study, gene expression profiles were used as expression traits. And relationships among the genes, which were co-linked to a common SNP(s), were identified by integrating the two types of information. Further research on the co-expressions among the co-linked genes was carried out after the gene-SNP relationships were established using the Haseman-Elston sib-pair regression. The results showed that the co-expressions among the co-linked genes were significantly higher if the number of connections between the genes and a SNP(s) was more than six. Then, the genes were interconnected via one or more SNP co-linkers to construct a gene-SNP intermixed network. The genes sharing more SNPs tended to have a stronger correlation. Finally, a gene-gene network was constructed with their intensities of relationships (the number of SNP co-linkers shared) as the weights for the edges.