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The spotted scat (Scatophagus argus) is an important ornamental species with sexually biased ornamental values that favor males. Therefore, it makes sense to breed mono-male fingerlings as ornamental fish. The spotted scat has an XX/XY sex determination system; therefore, the first step in producing genetically all-male offspring should be the induction of a fertile sex-reversed XY pseudo-female, which would then be mated with a normal XY male to produce a YY super-male. However, the XY pseudo-female produced by estradiol (E2) treatment failed due to ovarian malformations. Here, male to female sex reversal was induced in spotted scat through a combination of E2 and an androgen inhibitor (trilostane, TR). Spotted scat fingerlings of approximately 2.8–3.5 cm were fed a diet containing both E2 (300 µg/g) and TR (300 µg/g). The fish were treated for 90 days and then fed a normal diet until they reached one year of age, when they were sampled. Twenty-eight treated XY individuals were identified using sex-linked markers. According to their gonadal histological characteristics, these treated XY fish could be divided into three groups: males with testes (n = 21), intersex individuals with ovaries–testes (n = 3), and fully sex-reversed individuals with ovaries (n = 4). All treated XX fish (n = 8) developed into normal females. There were no obvious abnormalities in the ovaries of the XY sex-reversed fish compared to the treated XX fish. Serum 11-ketotestosterone (11-KT) levels were significantly lower in XY-reversed females and treated XX females than in XY males. Compared with XY male fish, the expression of female (42sp50, foxl2, figla, zar1, and zp2) and male (dmrt1, gsdf, amh and cyp11b2) biased genes was up- and down-regulated, respectively, in the gonads of XY-completely sex-reversed fish. Immunohistochemical results indicate that 42Sp50 was expressed in oocytes of XY-partially and completely sex-reversed fish, while strong Gsdf signals were mainly detectable in testicular somatic cells of XY-partially sex-reversed fish and XY male fish. XY females were successfully produced by the combined treatment of E2 and TR in the spotted scat. Their fertility will be tested in the future. [ABSTRACT FROM AUTHOR]

Hypothalamus are the important endocrine organs, which play key roles in regulating body growth, reproduction and hormone activity in other endocrine glands and organs. Estrogen, an important sex steroid hormone, is mainly synthesized and secreted in the gonads. It could regulate the reproduction and growth related genes expression in hypothalamus through feedback, and then participate in the regulation of corresponding physiological activities. Spotted scat (Scatophagus argus) is a good model to investigate the sexual growth dimorphism with a faster growth in females than males. The effect of 17β-estradiol (E2) injection in vivo on genes expression in hypothalamus of XY male spotted scat and the differences in genes expression between XY males and XX females remain unclear. In order to explore gene expression difference between XY male and XX female spotted scat hypothalamus and effect of in vivo injection in XY males, transcriptome analysis of hypothalamus were carried out, including sequencing data quality control, gene function annotation, differentially expressed gene (DEGs) screening and identification and pathway enrichment analysis, then real-time quantitative PCR (qPCR) was performed to detect genes expression. A total of 275833710 Clean reads were sequenced in this study, and the Q30 and GC value were more than 95% and 48%, respectively. In Ctrl-XX-H vs. Ctrl-XY-H group, 91 DEGs were screened and identified successfully, including 36 up-regulated and 55 down-regulated genes. In Ctrl-XX-H vs. E2-XY-H group, 28 DEGs were screened and identified, including 11 up-regulated and 17 down-regulated genes. GO and KEGG pathway analysis showed that DEGs was significantly enriched in biological processes such as cells, single cell, membrane, membrane components, and signal pathways such as ubiquinone and other terpene quinone biosynthesis, steroid hormone biosynthesis. By transcriptome and qPCR analyses, the expression of nrip1b, prl, hspa8b, snx21, cep63, ccna1, hsp90aa1.1, slc1a5, zar1, slc25a1b and loc124068175, were significantly different between XX and XY spotted scat, while the reproductive axis genes (sbGnRH, sGnRH and cGnRH) and growth axis genes (ghrh, sst1, sst3, sst5 and sst6) expression showed no significant differences. In XY male fish, six hours after E2 injection, the expression of cdc42ep1a, cyp19a1b, greb1, kank1a, snx21, and tmem30c in hypothalamus were significantly up-regulated, while prl expression was significantly down-regulated. Above results suggest that E2 might regulate some reproduction related genes expression in hypothalamus through feedback effects, which plays the critical roles in gametogenesis and reproduction regulation. [ABSTRACT FROM AUTHOR]

Marker‐5, which is linked to the major sex‐determination gene amhy on LG23, has been used for sex genotyping of various strains of Nile tilapia. Marker‐5 specifically amplifies fragments of approximately 1,422 and 982 bp of the X and Y chromosomes, respectively. Herein, we evaluated Marker‐5 polymorphism in Nile tilapia of Hainan strain, China. Agarose gel electrophoresis showed that in 13 out of 85 female fish, Marker‐5 amplified a larger band, distinct from the standard X‐specific band. Sanger sequencing showed that this larger band was 1,688 bp in length and showed high similarity with both 1,422‐bp X‐specific and 982‐bp Y‐specific fragments. The larger band was proved to be a female sex‐linked allele of Marker‐5 and was labeled as XL. When the polymerase chain reaction (PCR) products of XLY fish, amplified by Marker‐5, were tested by agarose gel electrophoresis, an unusual band, smaller than XL but larger than that of Y fragment, was observed. The unusual band could be a heteroduplex of the XL and Y fragments. In very few XXL male, amhy was found to be linked with 1,422‐bp fragment amplified by Marker‐5. Understanding the Marker‐5 is useful for its utilization in sex‐controlled breeding in farmed Nile tilapia. [ABSTRACT FROM AUTHOR]

A retrospective study was performed on 3 patients with cerebral venous sinus thrombosis (CVST) who were characterized with thalamic lesions in MRI examination. Among 3 CVST cases, one were male, and 2 was female, aged from 17 to 68. All cases had risk factors of CVST such as acute dehydration and puerperium. The main clinical symptoms included headache, nausea and vomiting, hypomnesis, hemiplegia and coma. MRI examination showed one case with unilateral thalamic lesions and 2 cases with bilateral thalamic lesions. Magnetic resonance venography (MRV) and DSA revealed straight sinus lesions in 3 cases, inferior sagittal sinus lesions in 2 cases, superior sagittal sinus lesion in one case, and transverse sinus lesion in one case, respectively. The patients with unilateral or bilateral thalamic lesions, especially those who had risk factors of CVST, should be diagnosed correctly and timely. [ABSTRACT FROM AUTHOR]

Objective: To investigate the current status of exclusive breastfeeding for the second child in the context of the universal two-child policy and the factors influencing exclusive breastfeeding., Methods: A self-designed questionnaire for the current status of breastfeeding and related factors influencing breastfeeding for the second child were used to survey 836 mothers with a second child, who were selected by cluster sampling, in Quzhou, Zhejiang, China., Results: A total of 680 usable questionnaires were obtained. The rate of exclusive breastfeeding for the second child was significantly lower than for the first child (34.9% vs 42.2%; P<0.05). The univariate analysis revealed that there were significant differences between the exclusive and non-exclusive breastfeeding groups in the mother′s age, education background, occupation and time of maternity leave, mode of delivery of the first child, sex of the first child, feeding pattern of the first child, mode of delivery of the second child, whether the second child was admitted to the intensive care unit, whether the father supported breastfeeding, and whether the grandmother/maternal grandmother supported breastfeeding (P<0.05). The multivariate logistic regression analysis showed that artificial feeding+partial breastfeeding for the first child (OR=12.286, P<0.05), cesarean section for the second child (OR=1.724, P<0.05), and having no breastfeeding support from the maternal grandmother (OR=1.651, P<0.05) were main factors for influencing exclusive breastfeeding., Conclusions: The current status of exclusive breastfeeding for the second child is not optimistic in the context of the universal two-child policy. Education about breastfeeding should be taken seriously at the birth of the first child, the rate of cesarean section should be reduced, and the family members should support exclusive breastfeeding, in order to improve the status of exclusive breastfeeding.

The CrylA Crystal Protein from Bacillus thuringiensis is associated with DNA, but the role and sequences of these DNA molecules are unknown. CrylA bipyramidal crystals from B. thuringiensis strain 4.0718 was selectively dissolved and associated DNA was extracted from protoxin. The DNA was digested with Nde I to obtain 3 to 5 kb fragments and then the fragments were subcloned into pMD18-T vector, screening of recombinants were done by PCR-RFLP and sequencing. The ORF of cry1Ac gene was amplified by primers designed and then subcloned. The 3.5 kb BamH I and Sal I fragments of pMDX35 was inserted into the pET30a vector, giving 8.9 kb recombinant plasmid, pETX35. ETX35 strain were obtained by transformed pETX35 into B121 (DE3). A 141 kD fusion protein was superexpressed as inclusion bodies. Quantitative protein analysis indicated that the amount of 141 kD protein was above the level of 51.36% of total cellular protein. Plasmid pHTX42 constructed from shuttle vector pHT304 was transformed B. thuringiensis acrystalliferous strain XBU001 with electroporation to obtain the recombinant HTX42. The recombinant protein was found with a molecular mass of 130 kD on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Scanning analysis indicated that the expressed protein accounted up to 79.28% of total cellular proteins and accumulated in the cells mounted up to 64.13% of cellular dry weight. Under Atomic Force Microscopy (AFM), typical bipyramidal crystals from HTX42 strain were found with a size of 1.2 microm x 2.0 microm. Bioassay showed that these inclusion bodies of ETX35 strain and crystals from HTX42 strain were highly toxic against the larvae of Plutella xylostella. On such a base, constructing insecticidal recombinant and analyzing the source, structure, and function of the 20 kb DNA can be further achieved.