The etiology of polycystic ovary syndrome (PCOS) is complex and the pathogenesis is not fully understood. Some studies have shown that dysregulation of ovarian granulosa cells may be related to abnormal follicles and excessive androgen in women with PCOS. Our team has also confirmed the high expression status of H19 in PCOS patients in the early stage. However, the relationship between H19 and miR-19b in the development of PCOS is still unknown. Therefore, we used bioinformatics to predict the binding sites of human H19 and miR-19b, and of miR-19b and CTGF genes. After the silencing and overexpression of H19, real-time polymerase chain reaction (PCR) was used to detect the expressions of H19, miR-19b, and CTGF. Western blotting was used to detect CTGF protein. Proliferation of KGN cells after H19 silencing was detected by CCK8. Flow cytometry was used to detect the apoptosis of KGN cells after H19 silencing. After the overexpression of H19, it was found that the expression of miR-19b gene decreased and the expression of CTGF increased, whereas silencing of H19 did the opposite. In addition, H19 could promote cell proliferation and decrease cell apoptosis. Finally, luciferase reporter assays showed that the 3′-end sequences of lncRNA H19 and CTGF contained the binding site of miR-19b. In conclusion, our study indicated that lncRNA H19 acted as a ceRNA to bind to miR-19b via a “sponge” to regulate the effect of CTGF on KGN cells, which may play a vital role in PCOS.

Abstract Background Cimicifuga racemosa is one of the herbs used for the treatment of climacteric syndrome, and it has been cited as an alternative therapy to estrogen. Apart from hectic fevers, dyspareunia and so on, dry mouth also increase significantly after menopause. It has not yet been reported whether C. racemosa has any impact on the sublingual gland, which may relate to dry mouth. In an attempt to determine this, we have compared the effects of estrogen and C. racemosa on the sublingual gland of ovariectomized rats. Results HE staining showed that the acinar cell area had contracted and that the intercellular spaces were broadened in the OVX (ovariectomized rats) group, while treatment with estradiol (E2) and iCR (isopropanolic extract of C. racemosa) improved these lesions. Transmission electron microscopy showed that rough endoplasmic reticulum expansion in mucous and serous acinar epithelial cells and apoptotic cells was more commonly seen in the OVX group than in the SHAM (sham-operated rats) group. Mitochondria and plasma membrane infolding lesions in the striated ducts were also observed. These lesions were alleviated by both treatments. It is of note that, in the OVX + iCR group, the volume of mitochondria in the striated duct was larger than in other groups. Immunohistochemical staining showed that the ratio of caspase-3 positive cells was significantly increased in the acinar cells of the OVX group compared with the SHAM group (p < 0.05); and the MA (mean absorbance) of caspase-3 in the striated ducts also increased (p < 0.05). Estradiol decreased the ratio of caspase-3 positive cells and the MA of caspase-3 in striated ducts significantly (p < 0.05). ICR also reduced the ratio of caspase-3 positive cells and the MA in the striated ducts (p < 0.05), but the reduction of the MA in striated ducts was inferior to that of the OVX + E2 group (p < 0.05). Conclusion Both estradiol and iCR can inhibit subcellular structural damage, and down-regulate the expression of caspase-3 caused by ovariectomy, but their effects were not identical, suggesting that both drugs confer a protective effect on the sublingual gland of ovariectomized rats, but that the specific location and mechanism of action producing these effects were different.

Para investigar os efeitos da sanguinarina (SAN) em lesões induzidas em ratos por radiação aguda, 45 ratos foram aleatoriamente divididos em grupo controle, grupo 10 Gy e grupo SAN+10 Gy. Os ratos dos grupos 10 Gy e SAN+10 Gy foram expostos à radiação de raio-X simples com uma dose acumulada de 10 Gy. Aos ratos do grupo SAN+10 Gy administraram-se, intraperitonealmente, 2.5 mg/kg de peso de SAN antes da radiação. Aos 5 dias de exposição à radiação, sacrificaram-se 5 ratos de cada grupo e retiraram-se amostras do intestino delgado, pulmões, baço e fígado para exames histopatológicos. Comparando com o grupo 10 Gy, a doença por radiação foi claramente atrasada e atenuada no grupo SAN+10 Gy. A análise de sobrevivência mostrou diferença significativa entre os dois grupos de radiação (P<0.05) e o tempo de sobrevivência média foi de mais 3 dias no grupo SAN+10 Gy do que no grupo 10 Gy (7.21±0.19 vs 4.20±0.13, P<0.001). Danos induzidos nos órgãos por radiação, baseados em exames histopatológicos, foram reduzidos pelo pré-tratamento com SAN. As pontuações de classificação da patologia Chiu, um índice para os danos intestinais, foi significativamente menor no grupo SAN+10 Gy do que no grupo 10 Gy (2.77±0.48 vs 4.37±0.31, P<0.01). Resultado semelhante foi obtido na pontuação patológica do pulmão (1.67±0.21 vs 2.33±0.38, P<0.01). As nossas descobertas preliminares mostram que SAN protege os animais contra doenças induzidas pela radiação e danos agudos nos órgãos seguidos de morte animal.

To investigate the effects of sanguinarine (SAN) on acute radiation induced injury in mice, 45 mice were randomly divided into control, 10 Gy and SAN+10 Gy groups. Mice in the 10 Gy and SAN+10 Gy groups were exposed to single X-ray radiation with an accumulated dose of 10 Gy. Mice in the SAN+10 Gy group were administered intraperitoneally with 2.5 mg/kg body weight of SAN before radiation. Five days after radiation exposure, 5 mice from each group were sacrificed and samples of the small intestine, lung, spleen and liver were fixed for histopathological examinations. Compared with the 10 Gy group, radiation sickness was obviously delayed or attenuated in the SAN+10 Gy group. Survival analysis showed a significant difference between 2 radiation groups (P<0.05) and mean survival time was 3 days longer in the SAN+10 Gy group than in the 10 Gy group (7.21±0.19 vs. 4.20±0.13, P<0.001). Radiation-induced organ damage, based on histopathological examinations, was decreased by SAN pretreatment. Chiu’s pathology grading scores, which is an index of intestinal damage, was significantly lower in the SAN+10 Gy group than in the 10 Gy group (2.77±0.48 vs. 4.37±0.31, P<0.01). A similar result was obtained in the pathological score of lung (1.67±0.21 vs. 2.33±0.38, P<0.01). Our preliminary findings demonstrated that SAN protects animals against radiation-induced sickness and acute damage to organs and following animal death.

The genetic diversity and genetic structure of three Chinese silkworm species Bombyx mori L., Antheraea pernyi Guérin-Meneville and Samia cynthia ricini Donovan were comparatively assessed based on RAPD markers. At the species level, A. pernyi and B. mori showed high levels of genetic diversity, whereas S. cynthia ricini showed low level of genetic diversity. However, at the strain level, A. pernyi had relatively highest genetic diversity and B. mori had lowest genetic diversity. Analysis of molecular variance (AMOVA) suggested that 60% and 72% of genetic variation resided within strains in A. pernyi and S. cynthia ricini, respectively, whereas only 16% of genetic variation occurred within strains in B. mori. In UPGMA dendrogram, individuals of A. pernyi and B. mori formed the strain-specific genetic clades, whereas those of S. cynthia ricini were distributed in a mixed way. The implications of these results for the conservation and utilization in breeding programs of three silkworm species are discussed.