Exploring the clinical and cellular mechanisms of LncRNA-KCNQ1OT1/miR-29a-3p/SOCS3 molecular axis in cases of unexplained recurrent spontaneous abortion.

OBJECTIVE: The objective of this study is to explore the functions and mechanisms of the LncRNA-KCNQ1OT1/miR-29a-3p/SOCS3 molecular pathway in the context of unexplained recurrent spontaneous abortion (URSA). METHODS: We conducted qRT-PCR to assess the levels of LncRNA-KCNQ1OT1, miR-29a-3p, and SOCS3 in both abortion tissues from women who experienced URSA and healthy early pregnant women. A dual-luciferase assay was employed to investigate whether miR-29a-3p targets SOCS3. Furthermore, RNA IP and RNA Pull-Down assays were employed to confirm the interaction between KCNQ1OT1 and SOCS3 with miR-29a-3p. RNA FISH was used to determine the cellular localization of KCNQ1OT1. Additionally, trophoblast cells (HTR8/SVneo) were cultured and the CCK-8 assay was utilized to assess cell proliferation, while flow cytometry was employed to analyze cell apoptosis. RESULTS: Compared to abortion tissues obtained from healthy early pregnant individuals, those from women who experienced URSA displayed a notable downregulation of KCNQ1OT1 and SOCS3, accompanied by an upregulation of miR-29a-3p. Suppression of KCNQ1OT1 resulted in the inhibition of cell proliferation and the facilitation of apoptosis in HTR8/SVneo cells. Our findings suggest that KCNQ1OT1 may exert a regulatory influence on SOCS3 through a competitive binding mechanism with miR-29a-3p. Notably, KCNQ1OT1 exhibited expression in both the cytoplasm and nucleus, with a predominant localization in the cytoplasm. Furthermore, we observed a negative regulatory relationship between miR-29a-3p and SOCS3, as the miR-29a-3p mimic group demonstrated significantly reduced cell proliferation and an increased rate of apoptosis when compared to the negative control (NC mimic) group. Additionally, the SOCS3 Vector group exhibited a substantial improvement in proliferation capability and a marked reduction in the apoptosis rate in comparison to the NC Vector group. The miR-29a-3p mimic + SOCS3 Vector group demonstrated a remarkable enhancement in proliferation and a reduction in apoptosis when compared to the miR-29a-3p mimic group. CONCLUSION: The competitive binding of miR-29a-3p to LncRNA-KCNQ1OT1 appears to result in the elevation of SOCS3 expression, consequently fostering the proliferation of trophoblast cells while concomitantly suppressing apoptosis.

Calorie restriction remodels gut microbiota and suppresses tumorigenesis of colorectal cancer in mice.

Colorectal cancer (CRC) is one of the most common cancers worldwide and the consumption of a high-calorie diet is one of its risk factors. Calorie restriction (CR) slows tumor growth in a variety of cancers, including colorectal cancer; however, the mechanism behind this remains unknown. In the present study, CR effectively reduced the tumor volume and weight in a xenograft BALB/c male nude mouse model. In addition, tumor immunohistochemistry revealed that the CR group had significantly higher expression of Bax (P<0.001) and significantly lower levels of Bcl2 (P<0.0001) and Ki67 (P<0.001) compared with control group. Furthermore, data from 16S ribosomal (r)RNA sequencing implied that CR was able to reprogram the microbiota structure, characterized by increased Lactobacillus constituent ratio (P<0.05), with amelioration of microbial dysbiosis caused by CRC. Further receiver operating characteristic curves demonstrated that the bacteria Bacteroides [area under the curve (AUC)=0.800], Lactobacillus (AUC=0.760) and Roseburia (AUC=0.720) served key roles in suppression of CRC in the mouse model. The functional prediction of intestinal flora indicated 'cyanoamino acid metabolism' (P<0.01), 'replication initiation protein REP (rolling circle plasmid replication)' (P<0.01), 'tRNA G10 N-methylase Trm11' (P<0.01) and 'uncharacterized protein with cyclophilin fold, contains DUF369 domain' (P<0.05) were downregulated in CR group. These findings implied that CR suppressed CRC in mice and altered the gut microbiota.

Chronic Ethanol Consumption Induces Osteopenia via Activation of Osteoblast Necroptosis.

Chronic high-dose alcohol consumption impairs bone remodeling, reduces bone mass, and increases the risk of osteoporosis and bone fracture. However, the mechanisms underlying alcohol-induced osteoporosis are yet to be elucidated. In this study, we showed that excess intake of ethyl alcohol (EtOH) resulted in osteopenia and osteoblast necroptosis in mice that led to necrotic lesions and reduced osteogenic differentiation in bone marrow mesenchymal stem cells (BMMSCs). We found that EtOH treatment led to the activation of the RIPK1/RIPK3/MLKL signaling, resulting in increased osteoblast necroptosis and decreased osteogenic differentiation and bone formation both in vivo and in vitro. We further discovered that excessive EtOH treatment-induced osteoblast necroptosis might partly depend on reactive oxygen species (ROS) generation; concomitantly, ROS contributed to necroptosis of osteoblasts through a positive feedback loop involving RIPK1/RIPK3. In addition, blocking of the RIPK1/RIPK3/MLKL signaling by necrostatin-1 (Nec-1), a key inhibitor of RIPK1 kinase in the necroptosis pathway, or antioxidant N-acetylcysteine (NAC), an inhibitor of ROS, could decrease the activation of osteoblast necroptosis and ameliorate alcohol-induced osteopenia both in vivo and in vitro. Collectively, we demonstrated that chronic high-dose alcohol consumption induced osteopenia via osteoblast necroptosis and revealed that RIPK1 kinase may be a therapeutic target for alcohol-induced osteopenia.

miR-100 promotes the proliferation of spermatogonial stem cells via regulating Stat3.

Micro RNAs play important roles during mammalian spermatogenesis, but the function(s) of specific miRNAs remain largely unknown. Here, we report that miR-100 is predominantly expressed in undifferentiated murine spermatogonia, including spermatogonial stem cells (SSCs). We utilized a miRNA mimic and inhibitor to knock down and overexpress miR-100 in cultured SSCs, respectively, finding that the miR-100 promotes the proliferation of SSCs in vitro. Furthermore, signals promoting SSC maintenance induced, whereas retinoic acid repressed, expression of miR-100. Stat3 expression was modulated by miR-100, with increased transcript and protein abundance in the presence of the miR-100 inhibitor versus reduced protein levels following miR-100 overexpression. Stat3 silencing also mimicked the reduced SSC proliferation phenotype associated with elevated miR-100 levels. Importantly, Stat3 silencing rescued the anti-proliferation capacity of miR-100 inhibitor on cultured SSCs. Given that the Stat3 3' untranslated region was not repressed by pre-miR-100 in a standard luciferase reporter assay, we suggest that miR-100 promotes SSC proliferation by indirect regulation of Stat3.

[Effect of Qilin Pill Combined Metformin on Polycystic Ovaries Induced Infertility Patients].

Objective To observe the effect and clinical efficacy of Qilin Pill (QP) combined met- formin on matrix metalloproteinase 9 (MMP-9), vascular endothelial growth factor (VEGF) , hepatocyte growth factor (HGF) , sex hormones, insulin resistance (IR) related indicators in polycystic ovaries induced infertility women. Methods Totally 58 polycystic ovarian syndrome (PCOS) complicated infertility patients admitted to authors' hospital were serial numbered according to visit sequence. The odd num- bers were recruited as the control group, and the even numbers were recruited as the test group, 29 ca- ses in each group. Patients in the control group took Metformin Hydrochloride Tablets, while those in the test group additionally took QP. All patients received 3 months of treatment. After treatment serum con- tents of VEGF, MMP-9, HGF, sex hormones, IR related indicators, and clinical efficacy were detected in all patients. Results (1) There was no statistical difference in serum contents of MMP-9, VEGF, HGF, sex hormones [luteinizing hormone (LH) , testosterone (T) ] , or serum levels of fasting serum insulin (FINS) and insulin sensitive index (IS]) related indicators between the two groups before treatment (P> 0. 05) ; (2) Compared with before treatment in the same group, there was statistical difference in serum contents of MMP-9, VEGF, HGF, and levels of LH, T, FINS, ISI in the two groups after treatment (P < 0. 05) ; (3) Compared with the control group, patients' serum contents of MMP-9, VEGF, HGF,and levels of LH, T, FINS, ISI were lower in the test group after treatment (P <0. 05) ; (4) After treatment the ovulation rate (93.2%) and the pregnancy rate (48.3%) were higher in the test group than in the control group (P <0.05). Conclusion QP combined metformin could significantly reduce serum levels of LH. FINS and T, contents of VEGF, HGF, and MMP-9, improve IR, and elevate the ovulation rate and the pregnancy rate in PCOS induced infertility patients.

Potential Paths for the Hydrogen-Bond Relaxing With (H2O)N Cluster Size.

Relaxation of the inter- and intra-molecular interactions for the hydrogen bond (O:H-O) between undercoordinated molecules determines the unusual behavior of water nanodroplets and nanobubbles. However, probing such potentials remains unreality. Here we show that the Lagrangian solution [Huang et al., J. Phys. Chem. B, 2013. 117: 13639] transforms the observed H-O bond (x = H) and O:H nonbond (x = L) lengths and phonon frequencies (dx, x) [Sun et al., J. Phys. Chem. Lett., 2013. 4: 2565] into the respective force constants and bond energies (kx, Ex) and hence enables the mapping of the potential paths for the O:H-O bond relaxing with water cluster size. Results show that molecular undercoordination not only reduces the molecular size (dH) with enhanced H-O energy from the bulk value of 3.97 to 5.10 eV for a H2O monomer, but also enlarges the molecular separation (dL) with reduced O:H energy from 95 to 35 meV for a dimer. The H-O energy gain raises the melting point from bulk value 273 to 310 K for the skin and the O:H energy loss lowers the freezing temperature from bulk value 258 to 202 K for 1.4 nm sized droplet, by dispersing the quasisolid phase boundaries.

[Effect of ruji recipe on the post-surgical survival of female breast cancer patients].

OBJECTIVE: To observe the effect of Ruji Recipe (RR) in preventing disease recurrence/metastasis and improving quality of life (QOL) for female breast cancer patients after operation. METHODS: Totally 102 female patients with stage I - III breast cancer were retrospectively analyzed. They were assigned to the treatment group (54 cases) and the control group (48 cases) according to whether they would rather accept RR therapy. Estrogen receptor/progesterone receptor (ER/PR) positive patients also accepted endocrine therapy. The overall survival (OS), disease-free survival (DFS), recurrence and metastasis, and QOL were compared between the two groups. RESULTS: Totally 100 patients completed the study. The median follow-up was 59 months. The median OS was 60 months in the treatment group and 52.5 months in the control group (chi2 = 3.274, P > 0.05). The median DFS was 55.0 months in the treatment group and 47.5 months in the control group (chi2 = 10.145, P < 0.01). The DFS rate was 75.9% (41/54) in the treatment group and 54.3% (25/46) in the control group (chi2 = -2.259, P < 0.05). There was statistical difference in the 2-, 3-, and 5-year DFS between the two groups (P < 0.01). There was statistical difference in the 2-year DFS 3-year DFS between stage II and III and stage III (P < 0.05, P < 0.01). There was statistical difference in the ER positive patients between 2-year DFS and 3-year DFS (P < 0.01, P < 0.05). There was statistical difference in the 3-and 5-year distant metastasis rate (DMR) in the treatment group, lower than that of the control group (3.7% vs 31.0%, 20.7% vs 60.7%; P < 0.01). By the end of follow-up, disease progression occurred in 13 cases of the treatment group, local recurrence in 3 cases, single organ metastasis in 7 cases, multi-metastasis in 3 cases, while the corresponding numbers were 21, 1, 11, and 9 in the control group (P < 0.05). As for 1 week before study and at 2-year follow-up using Functional Assessment of Cancer Therapy for Breast Cancer (FACT-B) system, there was statistical difference in the QOL between the two groups (P < 0.05), and better effect was obtained in the treatment group. CONCLUSION: RR, as an assistant therapy, could improve the OS rate, the DFS rate, and the QOL for post-surgical female breast cancer patients in 2 -3 years.

[Analysis of the automatic brightness controlling for x-ray imaging systems].

In this paper, the question about automatic brightness control for x-ray imaging systems based on CCD camera is discussed, and the structure and principle of an auto brightness control loop are analyzed along with the working procedure of the x-ray imaging system. A kind of digital brightness controller about a typical device and the designing idea of the computer brightness intelligent control software is introduced.