Knockdown of circ-FURIN suppresses the proliferation and induces apoptosis of granular cells in polycystic ovary syndrome via miR-195-5p/BCL2 axis

Background Polycystic ovary syndrome (PCOS) is an endocrine disease that increases the risk of infertility. Circular RNAs (circRNAs) play important roles in regulating the biological processes of PCOS. Our study was designed to explore the function of circ-FURIN in PCOS. Methods Circ-FURIN expression was detected using RT-qPCR. The protein expression of AVEN, BCL2, XIAP and AREL1 was measured using western blot. Dual-luciferase reporter and RNA pull-down assays were applied to clarify the interaction between miR-195-5p and circ-FURIN or BCL2. Functionally, cell proliferation was assessed by MTT and colony formation assays. Cell apoptosis was analyzed by flow cytometry. Results Circ-FURIN was upregulated in PCOS patients and granular cells (GCs). Knockdown of circ-FURIN inhibited cell proliferation and promoted apoptosis of KGN cells, along with the increased expression of caspase-3 and Bax and the decreased levels of p-PI3K. Gene ontology (GO) analysis indicated circ-FURIN is associated with apoptotic signaling pathway and cell death. Subsequently, BCL2 expression was elevated in patients with PCOS and positively regulated by circ-FURIN. Furthermore, circ-FURIN was served as a sponge of miR-195-5p to directly target to BCL2. The levels of miR-195-5p were reduced in PCOS and KGN cells. Knockdown of circ-FURIN decreased the expression of BCL2, which was abolished by miR-195-5p inhibitor. At last, rescue experiments revealed that overexpression of BCL2 reversed the effects of circ-FURIN knockdown on cell proliferation and apoptosis of KGN cells. Conclusions Loss of circ-FURIN alleviated the development of PCOS via miR-195-5p/BCL2 axis. Circ-FURIN may be the novel biomarker for PCOS.

A Novel Supplement Attenuates Oxidative Stress-Induced TDP-43-Related Pathogenesis in TDP-43-Expressed Cells

Amyotrophic lateral sclerosis (ALS) is caused by selective the loss of spinal motor neurons by multifactorial pathological mechanisms and results in muscle atrophy. Incidence rates of ALS are increasing over time, but there are no effective treatments at present due to limitations on approved therapies (riluzole and edaravone). Therefore, this study investigated whether combined treatment with Bojungikgi-tang and riluzole could act synergistically in transactive response DNA-binding protein 43 (TDP-43) stress granule cells. To examine the effect of combined treatment on oxidative stress-induced cell death, the CCK8 assay was performed for the detection of cell viability. The expression of oxidative stress-induced proteins was determined by Western blot. Quantification of sodium arsenite-induced reactive oxygen species (ROS) was measured in TDP-43 stress granular cells using 2,7-diacetyl dichlorofluorescein diacetate. To investigate the effect of combined treatment on TDP-43 aggregation, immunofluorescence and immunoblotting were performed in TDP-43 stress granular cells. This combined treatment alleviated oxidative stress-induced cell death by increasing the expression levels of antioxidation proteins, such as heme oxygenase-1 and B cell lymphoma-2-associated X protein. Furthermore, it reduced oxidative stress-induced TDP-43 aggregates and lowered the levels of autophagy-related proteins, including p62, light chain 3b, and ATG8, in TDP-43-expressing cells. Our results suggest that this combined treatment could be helpful for autophagy regulation in other neurodegenerative diseases.

Limosilactobacillus fermentum SWP-AFFS02 Improves the Growth and Survival Rate of White Shrimp via Regulating Immunity and Intestinal Microbiota

White shrimp Litopenaeus vannamei is an important species of farmed shrimp. Intestinal bacterial composition and immune activity play important roles in regulating the health condition of shrimp. Lactic acid bacteria Limosilactobacillus fermentum SWP-AFFS02 was isolated from the intestine of sea fish Rachycentron canadum, and the potential of its effect on growth, immunity, and intestinal microbiota of L. vannamei shrimp was investigated. Shrimps received feed with or without the addition of 8 log CFU/g L. fermentum SWP-AFFS02 thrice a day for 8 weeks. After 8-week treatment, weight gain, feed conversion rate, and survival rate of shrimp were greater in the L. fermentum SWP-AFFS02-feed group than in the control group. L. fermentum SWP-AFFS02 treatment increased the number of granular cells and semi-granular cells and decreased hyaline cell number when compared to the control group. L. fermentum SWP-AFFS02 promoted prophenoloxidase (PO) activity through increasing immune-associated gene expression in the hepatopancreas of shrimp. In addition, administration of feed containing L. fermentum SWP-AFFS02 regulated intestinal microbiota via decreasing the ratio of pathogenic bacteria, such as Vibrionaceae and Enterobacteriaceae, in the intestine of shrimp. This study demonstrated that administration of L. fermentum SWP-AFFS02 effectively prevented infection of L. vannamei shrimp by regulating intestinal microbiota and enhancing immunity in shrimp to increase the growth and improve their health status, which acted as a probiotic and provided beneficial effects on shrimp.

Haemocytes during Different Stages of Lifecycle in Bombyx mori (L.)

Haemocytes in insects mediates the cellular responses like phagocytosis, encapsulation and clotting which signifies the immunological functions of any insect. In the domesticated variety of silkworm B.mori five types of haemocytes have been identified in all the stages. The most abundant cell was found to be plasmatocytes followed by Granular cells. Spherule cells and Oenocytoids were found to less during IV and V instars and least or absent in all other stages. Haemocytes found be to fluctuating before and after spinning. Plasmatocytes and granular cells decreased gradually with respect to number of days in pupa. The total haemocytic count increases gradually in the silkworm larval stages and found to be maximum at the last instar and least during adult stages where the role of haemocytes is not required because they die after laying eggs.

Effect of Majapahit (Crescentia cujete L.) fruit powder on the immune profile of Litopenaeus vannamei after infection with Vibrio spp.

Background and Aim: The use of bioactive compounds is a promising tool to improve shrimp health regarding vibriosis. This study aimed to determine the effect of the dietary Majapahit (Crescentia cujete L.) fruit powder on the cellular immune response of vannamei shrimp (Litopenaeus vannamei) infected with Vibrio harveyi, Vibrio alginolyticus, and Vibrio parahaemolyticus. Materials and Methods: Twelve vannamei shrimp (aged 1 month) were randomly divided into four groups (n=3). Vannamei shrimp received experimental feed with different concentrations of Majapahit fruit powder for 20 days: Group A, 0%; Group B, 3.04%; Group C, 6.08%, and Group D, 9.12%. Subsequently, vannamei shrimp was infected with V. harveyi, V. alginolyticus, and V. parahaemolyticus. The total hemocytes, total differential hemocytes (hyaline, semi-granular, and granular cells), and phagocytic activity were assessed. Data were analyzed through analysis of variance (p<0.05) using SPSS ver. 24 for Windows. Results: Majapahit fruit powder at a dose of 3.04% increased the number of hyaline cells of L. vannamei after infection with Vibrio spp. Supplementation of the feed formula with Majapahit fruit powder at a dose of 3.04% increased the number of semi-granular and granular cells compared with the control. Furthermore, Majapahit fruit powder at doses of 3.04% and 6.08% increased the hemocytes compared with a dose of 9.12%. The phagocytic activity of L. vannamei after infection with Vibrio spp. tends to increase after supplementation with 3.04% Majapahit fruit powder. Conclusion: This study showed that the addition of C. cujete L. powder to the feed formula increased the cellular immune response. The most effective dose ranged from 3.04% to 6.08%.

The Different Sensitivity of Dendritic Degeneration in Hippocampal and Cortical Neurons to Estrogen Decline in Female Aging Mice

Aging is the most important risk factor for Alzheimer's disease (AD). Epidemiological studies reported that women have higher incidence of AD than men, which is associated with estrogen deficiency post menopause. The integrity of dendrites ensures normal neuronal function, and dendrite degeneration is one of the hallmarks in AD. However, the contribution of estrogen deficiency in dendritic remodeling during female aging is still unclarified. In the present study, female 18- and 22-month-old mice showed an age-dependent cognitive decline. Interestingly, female 18- and 22-month-old mice induced dendritic degeneration in both hippocampus and cortex, wherein the dendritic degeneration of hippocampal CA1 pyramidal neurons and cortical nonpyramidal neurons were more obvious in 18-month-old mice and that would not deteriorate in 22-month-old mice. The female mice after ovariectomy (OVX) 5 months induced similar changes in female 22-month-old mice indicating by cognitive decline and dendritic degeneration, however, dendritic degeneration of the hippocampal DG granular cells was less bad than that in female 22-month-old mice. Besides, estradiol (E2) level, and estrogen receptor α and β (ERα and ERβ) in hippocampus and cortex decreased in aged female and OVX mice. Importantly, E2 application rescued these changes induced by OVX. In conclusion, the female aging-induced dendritic remodeling existed regional differences, wherein hippocampal CA1 pyramidal neurons and cortical nonpyramidal neurons were the more vulnerable to onset of estrogen deficiency, while DG granular cells were more sensitive to age.