RNA reading protein YTHDF2 mediates Benzo(k)fluoranthene induced male reproductive injury by regulating the stability of BCL2.

Benzo (k) fluoranthene (BkF) has adverse effects on male reproduction, but its specific mechanism of action is still unclear. This study focused on the role of RNA reading protein YTHDF2 and its mechanism in BkF induced male reproductive injury. Mouse GC-2 spermatocytes were exposed to 0, 40, 80, 160 µM BkF. It was found that BkF significantly increased the apoptosis of GC-2 cell and decreased its survival rate. BCL2 in spermatocytes decreased significantly, while the expression of P53 and BAX exhibited a notable increase. Interestingly, the expression of RNA reading protein YTHDF2 progressively rose in tandem with the escalating BkF exposure dosage. Overexpression of YTHDF2 significantly reduced the viability of cells and increased the apoptosis rate. Meanwhile, there was a substantial increase in the expression of P53 and BAX, BCL2 was significantly down-regulated. On the contrary, interfering with YTHDF2 increased cell proliferation and reduced cell apoptosis. Furthermore, YTHDF2 overexpression exacerbated the decrease in cell viability under BkF exposure, while YTHDF2 knockdown was the opposite. The results from the RIP assay demonstrated a significant enhancement in the interaction of YTHDF2 protein with BCL2 mRNA following the overexpression of YTHDF2. In addition, animal experiments showed that there was an increase in apoptosis and a decrease in proliferation of testicular cells in mice in the high-dose (30 mg/kg) BkF group by TUNEL staining and Ki67 staining. Immunohistochemical analysis showed that BCL2 levels were significantly lower in the high-dose group than in the control group, while YTHDF2, P53 and BAX were dramatically increased. In summary, our study suggests that YTHDF2 has been implicated in BkF-induced male reproductive injury by promoting the degradation of BCL2.

FTO mediates bisphenol F-induced blood-testis barrier impairment through regulating ferroptosis via YTHDF1/TfRc and YTHDF2/SLC7A11 signal axis.

Bisphenol F (BPF) has been extensively utilized in daily life, which brings new hazards to male reproductive health. However, the specific functional mechanism is still unclear. Both cell and animal models were utilized for exploring the role of RNA methylation and ferroptosis and its underlying mechanisms in male reproductive injury induced by BPF. In animal model, BPF severely destroyed the integrity of the blood-testis barrier (BTB) and induced ferroptosis. Furthermore, BPF significantly affected the barrier function of TM4 cells and promoted ferroptosis. Importantly, ChIP assays revealed that BPF inhibited AR transcriptional regulation of FTO and FTO expression was downregulated in TM4 cells. Overexpression of FTO prevented the impairment of BTB by inhibiting ferroptosis in TM4 cells. Mechanistically, FTO could significantly down-regulate the m6A modification level of TfRc and SLC7A11 mRNA through MeRIP experiment. RIP experiments showed that YTHDF1 can bind to TfRc mRNA and promote its translation while YTHDF2 could bind to SLC7A11 mRNA and reduce its mRNA stability. Therefore, our results suggest that FTO plays a key role in BPF induced male reproductive toxicity through YTHDF1-TfRc axis and YTHDF2-SLC7A11 axis and may provide new ideas and methods for the prevention and treatment of male reproductive diseases associated with environmental pollutants.

Benzo(a)pyrene promotes the malignant progression of malignant-transformed BEAS-2B cells by regulating YTH N6-methyladenosine RNA binding protein 1 to inhibit ferroptosis.

Benzo[a]pyrene (BaP) is associated with the development of lung cancer, but the underlying mechanism has not been completely clarified. Here, we used 10 µM BaP to induce malignant transformation of human bronchial epithelial BEAS-2B cells, named BEAS-2B-T. Results indicated that BaP (6.25, 12.5 and 25 µM) treatment significantly promoted the migration and invasion of BEAS-2B-T cells. Meanwhile, BaP exposure inhibited ferroptosis in BEAS-2B-T, ferroptosis-related indexes Fe2+, malondialdehyde (MDA), lipid peroxidation (LPO) and reactive oxygen species (ROS) decreased significantly. The protein level of ferroptosis-related molecule transferrin receptor (TFRC) decreased significantly, while solute carrier family 7 membrane 11 (SLC7A11), ferritin heavy chain 1 (FTH1) and glutathione peroxidase 4 (GPX4) increased significantly. The intervention of ferroptosis dramatically effected the migration and invasion of BEAS-2B-T induced by BaP. Furthermore, the expression of YTH N6-methyladenosine RNA binding protein 1 (YTHDF1) was markedly increased after BaP exposure. YTHDF1 knockdown inhibited BEAS-2B-T migration and invasion by promoting ferroptosis. In the meantime, the contents of Fe2+, MDA, LPO and ROS increased significantly, TFRC was markedly increased, and SLC7A11, FTH1, and GPX4 were markedly decreased. Moreover, overexpression of YTHDF1 promoted BEAS-2B-T migration and invasion by inhibiting ferroptosis. Importantly, knockdown of YTHDF1 promoted ferroptosis and reduced BEAS-2B-T migration and invasion during BaP exposure, and overexpression of YTHDF1 increased migration and invasion of BEAS-2B-T by inhibiting ferroptosis during BaP exposure. RNA immunoprecipitation assays indicated that the binding of YTHDF1 to SLC7A11 and FTH1 markedly increased after YTHDF1 overexpression. Therefore, we concluded that BaP promotes the malignant progression of BEAS-2B-T cells through YTHDF1 upregulating SLC7A11 and FTH1 to inhibit ferroptosis. This study reveals new epigenetic and ferroptosis markers for preventing and treating lung cancer induced by environmental carcinogens.

Germplasm resources of three wood plant species enriched with nervonic acid.

Nervonic acid (NA) is a very-long-chain monounsaturated fatty acid with pharmaceutical and nutraceutical functions that plays an important role in treating several neurological disorders. One major source of NA is plant seed oil. Here we report fatty acid profiles of seeds and germplasm diversity of six plant species, including three woody plants with high amounts of NA-enriched seed oil, Malania oleifera, Macaranga adenantha, and M. indica. M. oleifera had the largest seed (average 7.40 g single seed), highest oil content (58.71%), and highest NA level (42.22%). The germplasm diversity of M. oleifera is associated with its habitat but not elevation. Seeds of M. adenantha contained higher NA levels (28.41%) than M. indica (21.77%), but M. indica contained a significantly higher oil content (29.22%) and seed yield. M. adenantha germplasm varied among populations, with one population having seeds with high oil content (22.63%) and NA level (37.78%).Although M. indica grow naturally at a range of elevations, no significant differences were detected between M. indica populations. These results suggest that M. indica and M. oleifera have greater potential as a source of NA, which will contribute to constructing a germplasm resource nursery and establishing a selection and breeding program to improve the development of NA-enriched plants.

Diagnostic re-evaluation and predictors of congenital hypothyroidism with eutopic thyroid gland in Jiangxi, China.

OBJECTIVES: An increase in the incidence of congenital hypothyroidism (CH) with eutopic gland has been reported worldwide due to neonatal screening programs. In this study, we aimed to determine the prevalence of transient CH (TCH) and to investigate predictive factors that could distinguish between permanent and transient CH in patients with eutopic thyroid glands. METHODS: We retrospectively reviewed 508 children treated for CH with eutopic thyroid glands between June 1998 and June 2020 in Jiangxi Newborn Screening Center. All patients were treated with levothyroxine and underwent Diagnostic re-evaluation after 2-3 years of age. Patients were classified as having TCH or permanent CH (PCH) during follow-up. RESULTS: Of the 508 patients initially treated for CH with a normally located gland, 335 patients (65.9%) were classified in the TCH group and 173 (34.1%) in the PCH group based on the defined criteria. Multivariate analysis revealed that TCH was associated with a lower levothyroxine dose at 24 months of age (p<0.001) and a lower likelihood of having a first-degree family history of CH (p=0.026) than PCH. Gender, prematurity, low birth weight, initial CH severity such as serum TSH and FT4 levels, or bone maturation delay at diagnosis had no effect. Receiver operating characteristics curve analysis showed that a cutoff of 2.3 µg/kg/day for levothyroxine dose requirement at 24 months of age had a sensitivity of 71% and a specificity of 70% for predicting transient CH, with values below this threshold considered predictive of transient CH. CONCLUSIONS: TCH presents a significant portion of patients with CH. The levothyroxine dose requirement at 24 months of age has a predictive role in differentiating TCH from PCH in CH patients with eutopic thyroid glands.

[Field application of antibody detection in a low transmission area of Schistosoma japonicum].

OBJECTIVE: To evaluate the field application of IHA and ELISA for schistosomiasis japonica detection at low transmission status. METHODS: 728 and 799 persons were examined by Kato-Katz's method, IHA and ELISA for schistosomiasis in an endemic village in the year of 2008 and 2010, respectively. The results of IHA and ELISA was evaluated in comparison to that of Kato-Katz (27 slides with 3 stool specimens) used as gold standard. RESULTS: The positive rate of Kato-Katz's method, IHA and ELISA were 10.3% (75/728), 40.0% (291/728) and 40.1% (292/728) in 2008, and 3.8% (30/799), 31.5% (252/799) and 40.1% (320/799) in 2010 respectively, in which significant difference was observed for the result between Kato-katz's method (chi2 = 26.92, P < 0.05) and IHA (chi2 = 11.82, P < 0.05). The consistency between the result of antibody detection and that of Kato-Katz's method was poor, lower than 0.2 (P < 0.01). If routine screening diagnosis mode was adopted, namely, population screened with IHA or ELISA first and confirmed with Kato-Katz's method, correlation analysis showed that the positive rate of Kato-Katz's method increased with the number of stool specimens and slides (rIHA2008 = 0.922, rELISA2008 = 0.908, rIHA2010 = 0.749, rELISA = 0.798; P < 0.05) . Those with egg positive but missed by IHA or ELISA mainly were cases with low infection intensity. When EPG < or = 40, the rate of detection ranged from 66.1% (39/59) to 87.0% (20/23) with IHA, and 62.7% (37/59) to 100% (23/23) with ELISA. When EPG > 40, however, all cases could be detected with ELISA, but some missed with IHA. CONCLUSION: In low transmission areas, the determination of target population for chemotherapy should be based on the examination of nine slides per stool specimen by Kato-Katz's method after serological screening.

[Evaluation of targeting property for Kuikang colon targeted pellets].

OBJECTIVE: To evaluate colon targeting characteristic of Kuikang colon targeted pellets (KCP) with determination of residual baicalin and baicalein concentration in gastrointestinal tract (GIT). METHOD: The baicalin and baicalein were assayed by HPLC. The recovery differences of the drug between KCP and conventional pellets from GIT were investigated, three and six hours after administration. RESULT: The baicalin recovery of KCP (70%) from rat GIT was higher than that of CP (about 20%). Most of KCP were intact at 3 h after oral administration, and distributed in lower ileum. It indicated that release site of KCP was in lower ileum and colon. Six hours later, a small amount of baicalin was recovered in intestime, which showed that the release of baicalin from KCP was complete. CONCLUSION: The determination of residual baicalin in rat GIT was feasibility for evaluating KCP. The result confirmed KCP of colon targeting property.

[Research progress on oral prolonged-release preparation of traditional Chinese medicine].

This paper reviews the lastest progress on oral prolonged-release preparation of traditional Chinese medicine. Four materials, include component, effective parts, single drug, and compound drugs of traditiong Chinese medicine, have been used to produce oral prolonged-release preparation. The main contents are study of preparation and evaluation of in vitro release. There are also some research works on integrative evaluation, pharmacokinetics and pharmacological activity of the prolonged-release preparation. It believes that the study on oral prolonged-release preparation of traditional Chinese medicine will have good prospect.