[Progesterone-induced microRNA-1a inhibits the proliferation of endometrial epithelial cells].

The aim of the present study was to investigate the effects of progesterone (P4)-induced microRNA-1a (miR-1a) on the proliferation of endometrial epithelial cells (EECs) and the underlying mechanism. In vivo, following subcutaneous injection of estradiol (E2) alone (E2 group) or combined injections of E2 and P4 (E2P4 group) in ovariectomized mice, quantitative real-time PCR (qPCR) was used to check the expression of miR-1a-3p in the directly isolated mouse EECs. The agomir or antagomir specific for miR-1a-3p was injected into one side of the uterine horns of ovariectomized mice pretreated with E2 alone or in combination with P4, and the non-specific control agomir or antagomir was injected into their contralateral horns. Flow cytometry was used to analyze the cell cycle of EECs. Immunohistochemistry (IHC) was used to examine the location and expression of cyclin D2, cyclin E1, and cyclin E2 in the uterine tissue sections. In vitro, primary cultured mouse EECs were pretreated with E2 alone (E2 group) or in combination with P4 (E2P4 group). qPCR was used to detect the expression of miR-1a-3p. Exogenous mimic of miR-1a-3p was transfected into E2-pretreated EECs, and EdU incorporation analysis was used to test the proliferation activity of the EECs. The result of in vivo experiment showed that the expression of miR-1a-3p in E2P4 group was significantly higher than that in E2 group (P < 0.05). The miR-1a-3p agomir arrested cell cycle at G1 to S transition in the mice injected subcutaneously with E2 alone (P < 0.05). Conversely, silencing of miR-1a-3p with transfection of miR-1a-3p antagomir promoted the entry of cells into S phase in the mice injected subcutaneously with both E2 and P4 (P < 0.05). The expressions of cyclin E1 and cyclin E2, except for cyclin D2, in uterine sections were also dramatically reduced by miR-1a-3p overexpression in the uterine epithelium (P < 0.05). In vitro, miR-1a-3p was not expressed in the cells of both E2 and E2P4 groups. The mimic of miR-1a-3p decreased EECs proliferation activity (P < 0.05). These results indicate that P4-induced miR-1a can inhibit the expression of cyclin E1 and cyclin E2, consequently suppressing the proliferation of mouse EECs by arresting cells at G1/S phase.

[Effect of Progesterone-induced MicroRNA-152 on the Proliferation of Endometrial Epithelial Cells].

OBJECTIVE: To determine the expression of microRNA-152 induced by progesterone and its effect on the proliferation of endometrial epithelial cells (EECs). METHODS: Cultured EECs, Ishikawa were divided into four groups: control group (C group), 10(-8) mol/L estrogen treated group (E group), 10(-6) mol/L progesterone group (P group) and estrogen plus progesterone treated group (E&P group). The expression of mature microRNA-152 (microRNA-152-3p) of was detected by qRT-PCR. The estrogen treated cells were transfected with mimic-microRNA-152-3p. The estrogen and progesterone treated cells were transfected with inhibitor-microRNA-152-3p. Cell proliferations were detected by CCK-8 assay. The target gene of microRNA-152-3p proteins was predicted using microRNA target databases and validated by Western blot. RESULTS: qRT-PCR showed no difference between C and E groups (P > 0.05) in the expression of microRNA-152-3p. P group had higher expressions of microRNA-152-3p than C group (P < 0.05). E&P group had higher expressions of microRNA-152-3p than C group and P group. MicroRNA target protein prediction suggested that CDC14A is one of direct target proteins of microRNA-152-3p. The results of CCK-8 assay showed that mimic-microRNA-152-3p transfection blocked proliferations of estrogen treated cells and lowered expressions of CDC14A in these cells; while inhibitor-microRNA-152-3p promotes proliferations of estrogen and progesterone treated cells and increased expressions of CDC14A in these cells. CONCLUSION: Progesterone may suppress proliferations of EECs through inducing expressions of microRNA-152-3p. CDC14A is probably one target protein of microRNA-152-3p for its action on EECs.

A Report of Two Cases of Meningoencephalitis Caused by Streptococcus intermedius.

Objective: In this paper, we analyzed the clinical data of patients with meningoencephalitis caused by Streptococcus intermedius to understand better the clinical characteristics of the disease and recommend auxiliary diagnostic mode as well as treatment experience. Methods: We reviewed the clinical data of two patients admitted to our department in 2019 with meningoencephalitis caused by S. intermedius. Results: Two female patients were examined, one of whom had a history of radiotherapy for nasopharyngeal carcinoma while the other had no underlying disease. These two patients were admitted with symptoms of meningoencephalitis. Cerebrospinal fluid examinations revealed elevated levels of leukocytes and protein. After treatment with meropenem, the condition improved for a brief time, but then worsened with a decline in mental status and limb movement. Blood and cerebrospinal fluid cultures demonstrated the absence of pathogenic bacteria, while genome sequencing of cerebrospinal fluids revealed the presence of S. intermedius. Cranial magnetic resonance imaging revealed multiple cerebral abscesses (CAs). After coadministration of linezolid as an anti-infective, clinical symptoms gradually improved, and the CAs shrank on follow-up imaging. The condition exhibited a pattern of improvement-deterioration-improvement. Conclusion: Meningoencephalitis caused by S. intermedius is complex and prone to fluctuation and formation of multiple CAs. The definitive clinical diagnosis of this disease can be aided by genome sequencing technology, and early clarification of the etiology combined with the use of potent antibiotics is effective.

Progesterone-Induced miR-145/miR-143 Inhibits the Proliferation of Endometrial Epithelial Cells.

Our previous study showed that progesterone (P4) can specifically regulate the expression of some microRNAs (miRNAs) in endometrial epithelium. In the present study, we verified the P4-dependent expression of miR-145/miR-143 in endometrial epithelial cells, explored the regulative mechanism of the P4 receptor (PR), and investigated their effects on the proliferation of endometrial epithelial cells. Our results showed that P4 can induce the expression of miR-145/143 in endometrial epithelial cells by acting on the PR A subtype. P4-induced miR-145/143 can inhibit the expression of cyclin D2 by binding to cyclin D2 mRNA 3'UTR. It can also inhibit cell proliferation in mouse endometrial epithelium by arresting the cell cycle during the G1-S checkpoint. Furthermore, miR-145 and miR-143 can inhibit the proliferation of human endometrial cancer cells. In conclusion, P4-induced miR-145/miR-143 is an important regulator in the proliferation of endometrial epithelial cells, and it can also inhibit the proliferation of human endometrial cancer cells. Our study indicates miRNAs are important mechanism of P4 in inhibiting the proliferation of endometrial epithelial cells. And these miRNAs are potential candidates for the diagnosis of endometrial cancer and therapeutic targets.

Nanostructured Thermoresponsive Surfaces Engineered via Stable Immobilization of Smart Nanogels with Assistance of Polydopamine.

Thermoresponsive surfaces featured with nanostructures have found wide potential applications in biological and chemical fields. Herein, we report nanostructured thermoresponsive surfaces engineered via stable immobilization of thermoresponsive nanogels with the assistance of polydopamine. The results show that the thin layer of polydopamine on the poly( N-isopropylacrylamide) (PNIPAM) nanogels nearly does not affect the thermoresponsive property of the nanogels. The stability of the thermoresponsive nanogels on the substrate surfaces immobilized under different pH conditions of dopamine solutions are quatitively studied by fluid shearing experiments inside capillaries, and the characterization results show that the strong interaction forces between the polydopamine layer on the substrate surfaces and the thermoresponsive nanogels are heavily dependent on the oxidation state of the dopamine molecules. With the proposed strategy, thermoresponsive nanostructured surfaces immobilized with PNIPAM nanogels on two-dimensional and three-dimensional substrate surfaces are generated to achieve smart cell culture plates and smart gating membranes, respectively, which demonstrate versatile applications of the nanostructured thermoresponsive surfaces.

Progesterone-Induced miR-152 Inhibits the Proliferation of Endometrial Epithelial Cells by Downregulating WNT-1.

Progesterone (P4) is an important ovarian hormone that inhibits estrogen-dependent proliferation of endometrial epithelial cells (EECs). miR-152 has been reported to be a cell cycle regulator. In this study, we first demonstrated that P4 induced the expression of miR-152 in ovariectomized mice and Ishikawa cell. miR-152 was detected in the human endometrial cell lines that were stably transfected with P4 receptor. Results showed that P4 induced its expression through its receptor B subtype. Then, using the specific miRNA mimic and inhibitor, we proved that miR-152 impeded G1/S transition in the cell cycle of EECs and inhibited cellular proliferation via downregulating WNT-1 in mice and human endometrial cancer cell lines (Ishikawa, HEC-1-b, and KLE). miR-152 induced by P4 is an important inhibitor for the proliferation of EECs. miR-152 may be an important tumor suppressor microRNA in endometrial cancer.

Identification and characterization of progesterone- and estrogen-regulated MicroRNAs in mouse endometrial epithelial cells.

In endometrial epithelial cells, progesterone (P4) functions in regulating the cell structure and opposing the effects of estrogen. However, the mechanisms of P4 that oppose the effects of estrogen remain unclear. MicroRNAs (miRNAs) are important posttranscriptional regulators that are involved in various physiological and pathological processes. Whether P4 directly induces miRNA expression to antagonize estrogen in endometrial epithelium is unclear. In this study, total RNAs were extracted from endometrial epithelium of ovariectomized mice, which were treated with estrogen alone or a combination of estrogen and P4. MicroRNA high-throughput sequencing with bioinformatics analysis was used to identify P4-induced miRNAs, predict their potential target genes, and analyze their possible biological functions. We observed that 146 mature miRNAs in endometrial epithelial cells were significantly upregulated by P4. These miRNAs were extensively involved in multiple biological processes. The miRNA-145a demonstrated a possible function in the antiproliferative action of P4 on endometrial epithelial cells.