CircMAST1 inhibits cervical cancer progression by hindering the N4-acetylcytidine modification of YAP mRNA.

BACKGROUND: Cervical cancer (CCa) is the fourth most common cancer among females, with high incidence and mortality rates. Circular RNAs (circRNAs) are key regulators of various biological processes in cancer. However, the biological role of circRNAs in cervical cancer (CCa) remains largely unknown. This study aimed to elucidate the role of circMAST1 in CCa. METHODS: CircRNAs related to CCa progression were identified via a circRNA microarray. The relationship between circMAST1 levels and clinicopathological features of CCa was evaluated using the clinical specimens and data of 131 patients with CCa. In vivo and in vitro experiments, including xenograft animal models, cell proliferation assay, transwell assay, RNA pull-down assay, whole-transcriptome sequencing, RIP assay, and RNA-FISH, were performed to investigate the effects of circMAST1 on the malignant behavior of CCa. RESULTS: CircMAST1 was significantly downregulated in CCa tissues, and low expression of CircMAST1 was correlated with a poor prognosis. Moreover, our results demonstrated that circMAST1 inhibited tumor growth and lymph node metastasis of CCa. Mechanistically, circMAST1 competitively sequestered N-acetyltransferase 10 (NAT10) and hindered Yes-associated protein (YAP) mRNA ac4C modification to promote its degradation and inhibit tumor progression in CCa. CONCLUSIONS: CircMAST1 plays a major suppressive role in the tumor growth and metastasis of CCa. In particular, circMAST1 can serve as a potential biomarker and novel target for CCa.

The storage stability of Bacillus subtilis spore displaying cysteine protease of Clonorchis sinensis and its effect on improving the gut microbiota of mice.

Bacillus subtilis (B. subtilis) spore can serve as an ideal vehicle for expressing heterologous antigens, and elicit specific immune responses by oral administration. In previous studies, we successfully constructed the recombinant B. subtilis spores expressing cysteine protease of Clonorchis sinensis (C. sinensis, B.s-CsCP), and confirmed that oral administration of B.s-CsCP could elicit good protective immune responses in mice. In this study, Gram staining was used to observe the morphology of B.s-CsCP in different form, and the storage of liquid spores and lyophilized spores at different temperatures was compared. The mice were orally immunized with three different doses of spores (2×108, 1×109, and 5×109 CFU/day) for three times in total at biweekly interval. Then, antibody levels of mice were measured, the safety of spores was evaluated, and the changes of gut microbiota after oral gavage of spores (1×109 dose) were investigated. Results showed that B. subtilis was a typical Gram-positive bacterium, and its spore had good resistance to chemical dye. Liquid B. subtilis spores resuspended in sterile water could be stored for a long time at 4 °C or below, while lyophilized spores could be well stored even at RT and better at lower temperatures. Oral administration of B. subtilis spores to mice could stimulate both local mucosal and systemic immune responses in a dose-dependent manner without toxic side effects. Besides, beneficial bacteria producing butyrate such as Odoribacter were increased, while potential pathogens such as Escherichia-Shigella were decreased in mice intestine. Therefore, our work further confirmed that B. subtilis spores expressing CsCP could be a promising oral vaccine against C. sinensis with the advantages of stability, safety, easy storage, and promotion of intestinal health.Key Points• Recombinant CsCP B. subtilis spores could be easily preserved in either liquid or freeze-dried state.• Oral immunization of recombinant spores in mice could increase both local and system immune levels in a dose-dependent manner.• Oral administration of recombinant spores increased the number of beneficial bacteria and reduced the number of harmful bacteria in the intestinal tract of mice.

Acquirement of HRP conjunct IgG anti-IgMs from most widely cultured freshwater fishes in China and its immunoreactivity.

Until now, custom-made or commercial polyclonal antibody against only one kind of fish IgM limited application of the antibody. During our research on development of vaccine against infection of Clonorchis sinensis (C. sinensis) in several kinds of fish, we were conscious of the urgency of secondary antibody to evaluate immune effect and screen C. sinensis infection with immunological technology instead of labor-intensive and time-consuming squash or artificial digestion of fish flesh. So that, we purified IgM of grass carp, bighead carp, crucian carp, common carp and tilapia which were widely cultured freshwater fishes in most areas of China. On this basis, we generated HRP-conjunct rabbit IgG anti-fish IgMs with high titers. IgM of other freshwater fishes including oshima, yellow catfish, bream, silver carp and so on could be recognized by the IgG sensitively. Additionally, The ELISA detection displayed that the IgG could be more specific and sensitive than custom-made rabbit IgG anti-grass carp IgM. The acquirement of HRP-conjunct rabbit IgG anti-fish IgMs was the cornerstone for studying the immune system of teleost fish, developing immunoassay methods and evaluation of fish vaccine with more convenience.

Evaluation of immune response to Bacillus subtilis spores expressing Clonorchis sinensis serpin3.

Clonorchis sinensis (C. sinensis) is one of the most serious food-borne parasites, which can lead to liver fibrosis or cholangiocarcinoma. Effective measures for clonorchiasis prevention are still urgently needed. Bacillus subtilis (B. subtilis) is an effective antigen delivery platform for oral vaccines. Chonorchis sinensis serpin (CsSerpin) was proved to be potential vaccine candidates. In this study, CsSerpin3 was displayed on the surface of B. subtilis spore and recombinant spores were orally administrated to BALB/C mice. CsSerpin3-specific IgA levels in faecal, bile and intestinal mucous increased at 4-8 weeks after the first administration compared with those in control groups. The mucus production and the number of goblet cells in intestinal mucosa elevated in B.s-CotC-CsSerpin3 (CotC, coat protein of B. subtilis spore) spores treated group compared to those in blank control. No significant difference in the activities of glutamic-pyruvic transaminase/ alanine aminotransferase and glutamic oxalacetic transaminase/aspartate aminotransferase were observed between groups. There was no side effect inflammation and observable pathological damage in the liver tissue of mice after administration. Moreover, collagen deposition and Ishak score were statistically reduced in B.s-CotC-CsSerpin3 spores treated mice. In conclusion, B. subtilis spores displaying CsSerpin3 could be investigated further as an oral vaccine against clonorchiasis.

Oral delivery of Bacillus subtilis spores expressing Clonorchis sinensis paramyosin protects grass carp from cercaria infection.

Clonorchis sinensis (C. sinensis), an important fishborne zoonotic parasite threatening public health, is of major socioeconomic importance in epidemic areas. Effective strategies are still urgently expected to prevent against C. sinensis infection. In the present study, paramyosin of C. sinensis (CsPmy) was stably and abundantly expressed on the surface of Bacillus subtilis spores. The recombinant spores (B.s-CotC-CsPmy) were incorporated in the basal pellets diet in three different dosages (1 × 105, 1 × 108, 1 × 1011 CFU/g pellets) and orally administrated to grass carp (Ctenopharyngodon idella). The immune responses and intestinal microbiota in the treated grass carp were investigated. Results showed that specific anti-CsPmy IgM levels in sera, skin mucus, bile, and intestinal mucus, as well as mRNA levels of IgM and IgZ in the spleen and head kidney, were significantly increased in B.s-CotC-CsPmy-1011 group. Besides, transcripts levels of IL-8 and TNF-αin the spleen and head kidney were also significantly elevated than the control groups. Moreover, mRNA levels of tight junction proteins in the intestines of B.s-CotC-CsPmy-1011 group increased. Potential pathogenetic bacteria with lower abundance and higher abundances of candidate probiotics and bacteria associated with digestion in 1 × 1011 CFU/g B.s-CotC-CsPmy spores administrated fishes could be detected compared with control group. The amount of metacercaria in per gram fish flesh was statistically decreased in 1 × 1011 CFU/g B.s-CotC-CsPmy spores orally immunized group. Our work demonstrated that B. subtilis spores presenting CsPmy on the surface could be a promising effective, safe, and needle-free candidate vaccine against C. sinensis infection for grass carp.

Lesion distribution characteristics of deep infiltrating endometriosis with ovarian endometrioma: an observational clinical study.

BACKGROUND: To investigate the characteristics of deep infiltrating endometriosis (DIE) lesion distribution when associated with ovarian endometrioma (OEM). METHODS: The present study analyzed retrospective data obtained by the First Affiliated Hospital of Sun Yat-sen University, between June 2008 to June 2016. A total of 304 patients underwent laparoscopic surgery for complete removal of endometriosis by one experienced surgeon, and histological confirmation of OEM associated with DIE was conducted for each patient. Clinical data were recorded for each patient from medical, operative and pathological reports. Patients were then divided into two groups according to unilateral or bilateral OEM. Patients with unilateral OEM were subsequently divided into two subgroups according to OEM location (left- or right-hand side) and the diameter of the OEM (≤50 and > 50 mm). The distribution characteristics of DIE lesions were then compared between the groups. RESULTS: DIE lesions were widely distributed, 30 anatomical sites were involved. Patients with associated unilateral OEM (n = 184 patients) had a significantly increased number of DIE lesions when compared with patients with bilateral OEM (n = 120 patients; 2.76 ± 1.52 vs. 2.33 ± 1.34; P = 0.006). Compared with bilateral OEM with DIE, there was a higher rate of intestinal (39.1% vs. 18.3%; P < 0.01) and vaginal (17.4% vs. 6.7%; P < 0.01) infiltration by DIE lesions in unilateral OEM with DIE. The mean number of DIE lesions was not significantly correlated with the location or size of the OEM (2.83 ± 1.56 vs. 2.74 ± 1.53; P = 0.678; and 2.65 ± 1.42 vs. 2.80 ± 1.43; P = 0.518, respectively). There was no significant difference between the groups with OEM ≤50 mm and > 50 mm. CONCLUSION: Lesion distribution characteristics in women diagnosed with histologically proven OEM associated with DIE were frequently multifocal and severe.

Oral delivery of Bacillus subtilis spores expressing grass carp reovirus VP4 protein produces protection against grass carp reovirus infection.

Grass carp (Ctenopharyngodon idellus) hemorrhagic disease (GCHD), caused by grass carp reovirus (GCRV), has given rise to an enormous loss in grass carp industry during the past years. Up to date, vaccination remained to be the most effective way to protect grass carp from GCHD. Oral vaccination is of major interest due to its advantages of noninvasive, time-saving, and easily-operated. The introduction of oral vaccination has profound impact on aquaculture industry because of its feasibility of extensive application for fish in various size and age. However, the main challenge in developing oral vaccine is that antigens are easily degraded and are easy to induce tolerance. Bacillus subtilis (B. subtilis) spores would be an ideal oral vaccine delivery system for their robust specialty, gene operability, safety and adjuvant property. VP4 protein is the major outer capsid protein encoded by GCRV segment 6 (S6), which plays an important role in viral invasion and replication. In this study, we used B. subtilis spores as the oral delivery system and successfully constructed the B. subtilis CotC-VP4 recombinant spores (CotC-VP4 spores) to evaluate its protective efficacy in grass carp. Grass carp orally immunized with CotC-VP4 spores showed a survival rate of 57% and the relative percent survival (RPS) of 47% after the viral challenge. Further, the specific IgM levels in serum and the specific IgZ levels in intestinal mucus were significantly higher in the CotC-VP4 group than those in the Naive group. The immune-related genes including three innate immune-related genes (IL-4/13A, IL-4/13B, CSF1R), four adaptive immune-related genes (BAFF, CD4L, MHC-II, CD8), three inflammation-related genes (IL-1ß, TNF-α, TGF-ß) and interferon type I (IFN-I) related signaling pathway genes were significantly up-regulated in the CotC-VP4 group. The study demonstrated that the CotC-VP4 spores produced protection in grass carp against GCRV infection, and triggered both innate and adaptive immunity post oral immunization. This work highlighted that Bacillus subtilis spores were powerful platforms for oral vaccine delivery, and the combination of Bacillus subtilis spores with GCRV VP4 protein was a promising oral vaccine.

Immune response induced by oral delivery of Bacillus subtilis spores expressing enolase of Clonorchis sinensis in grass carps (Ctenopharyngodon idellus).

Clonorchiasis, caused by the consumption of raw or undercooked freshwater fish containing infective metacercariae of Clonorchis sinensisis (C.sinensis), remains a common public health problem. New effective prevention strategies are still urgent to control this food-borne infectious disease. The previous studies suggested Bacillus subtilis (B. subtilis) spores was an ideal vaccines delivery system, and the C.sinensis enolase (CsENO) was a potential vaccine candidate against clonorchiasis. In the current study, we detected CsENO-specific IgM levels by ELISA in sera, intestinal mucus and skin mucus in grass carps (Ctenopharyngodon idella) through oral administration with B. subtilis spores surface expressing CsENO. In addition, immune-related genes expression was also measured by qRT-PCR. Grass carps orally treated with B. subtilis spores or normal forages were used as controls. The results of ELISA manifested that specific IgM levels of grass carps in CsENO group in sera, intestine mucus and skin mucus almost significantly increased from week 4 post the first oral administration when compared to the two control groups. The levels of specific IgM reached its peak in intestine mucus firstly, then in sera, and last in skin mucus. qRT-PCR results showed that 5 immune-related genes expression had different degree of rising trend in CsENO group when compared to the two control groups. Our study demonstrated that orally administrated with B. subtilis spores expressing CsENO induced innate and adaptive immunity, systemic and local mucosal immunity, and humoral and cellular immunity. Our work may pave the way to clarify the exact mechanisms of protective efficacy elicited by B. subtilis spores expressing CsENO and provide new ideas for vaccine development against C. sinensis infection.

Oral delivery of Bacillus subtilis spores expressing cysteine protease of Clonorchis sinensis to grass carp (Ctenopharyngodon idellus): Induces immune responses and has no damage on liver and intestine function.

Clonorchis sinensis (C. sinensis) is a fish-borne trematode. Human can be infected by ingestion of C. sinensis metacercariae parasitized in grass carp (Ctenopharyngodon idella). For induction of effective oral immune responses, spores of Bacillus subtilis (B. subtilis) WB600 were utilized as vehicle to delivery CsCP (cysteine protease of C. sinensis) cooperated with CotC (B.s-CotC-CP), one of coat proteins, to the gastrointestinal tract. After routine culture of 8-12 h in LB medium, B. subtilis containing CotC-CsCP was transferred into the sporulation culture medium. SDS-PAGE, western blotting and the growth curve indicated that the best sporulation time of recombinant WB600 was 24-30 h at 37 °C with continuous shaking (250 rpm). Grass carp were fed with three levels of B.s-CotC-CP (1 × 106, 1 × 107, and 1 × 108 CFU g-1) incorporated in the basal pellets diet. The commercial pellets or supplemented with spores just expressing CotC (1 × 107 CFU g-1) were served as control diet. Our results showed that grass carp orally immunized with the feed-based B.s-CotC-CP developed a strong specific immune response with significantly (P < 0.05) higher levels of IgM in samples of serum, bile, mucus of surface and intestinal compared to the control groups. Abundant colonization spores expressing CsCP were found in hindgut that is conducive to absorption and presentation of antigen. Moreover, B. subtilis spores appeared to show no sign of toxicity or damage in grass carp. Our cercariae challenge experiments suggested that oral administration of spores expressing CsCP could develop an effective protection against C. sinensis in fish body. Therefore, this study demonstrated that the feed-based recombinant spores could trigger high levels of mucosal and humoral immunity, and would be a promising candidate vaccine against C. sinensis metacercariae formation in freshwater fish.

Early cervical cancer impact of peritoneal vaginoplasty combined with laparoscopic radical hysterectomy improved sexual function.

OBJECTIVE: We investigated the impact of laparoscopic radical hysterectomy (LRH) in combination with peritoneal vaginoplasty (PV) in improving sexual function after radical hysterectomy (RH) in patients with early cervical cancer. METHODS: A total of 79 patients with early-stage cervical cancer younger than 45 years were assigned to receive LRH in combination with PV (the LRH-PV group; n = 31) or LRH alone (the LRH group; n = 48). Other 40 healthy females were selected as controls (the control group). The sexual function was assessed with Female Sexual Functioning Index (FSFI). The FSFI scores and sexual function in the postoperative 1 year were compared between the LRH-PV and LRH groups, LRH-PV and control groups, and LRH and control groups, respectively. RESULTS: Patients with LRH-PV showed significantly higher scores in sexual satisfaction, lubrication, pain, and total score than those with LRH alone (P < 0.05) but were not statistically different in scores regarding sexual desire, arousal, and orgasm (P > 0.05). Healthy controls showed the highest in total scores and 6 domains among all subjects. In addition, the FSFI total scores in the LRH-PV group, LRH group, and LRH-PV + LRH group were significantly decreased compared with the control (P < 0.05). CONCLUSIONS: Peritoneal vaginoplasty to lengthen the vagina improves sexual function of patients with early cervical cancer receiving LRH in sexual satisfaction, lubrication, and pain.

Analyzing simultaneous positive expression of EZH2 and P53 protein to improve predictive value in cervical squamous cell carcinoma.

OBJECTIVE: The current study was undertaken to investigate the predictive value of simultaneous enhancer of zeste homolog 2 (EZH2) and P53 expression in lesions of patients with cervical squamous cell carcinoma. METHODS: Quantum dot double fluorescence staining was applied to detect EZH2 and P53 protein in biopsy tissue of 168 patients with cervical squamous cell carcinoma. The expression was classified into double positive (EZH2 and P53 were positively expressed), single positive (either EZH2 or P53 was positively expressed), and double negative (neither was positively expressed). The EZH2 and P53 expression, clinical stages of cervical cancer, lymph node metastasis, progression-free survival, and overall survival were analyzed. RESULTS: A relationship of EZH2 and P53 expression with the clinical stage of cervical cancer and lymph node metastasis was indicated. Simultaneous detection of both proteins could partly predict prognosis, consistent with previous studies' results. The current study demonstrated that the expression levels of EZH2 and P53 in tumor tissue and the proportion of cases with double-positive expression significantly increased with increasing clinical stages of cancer, also confirming up-regulated expression of EZH2 and P53 with increasing stages. These findings may suggest that the 2 proteins were involved in the development of cervical cancer, but the nature of their interaction is undefined. The increase of EZH2 and P53 expression in patients with lymph node metastasis indicated that they may be involved in metastasis of cervical cancer. CONCLUSION: Simultaneous positive EZH2 and P53 expression could improve the predictive value of a poor prognosis in cervical cancer.

[Clinical significance of expression of enhancer of zeste homolog 2 and 53 in cervical squamous cell carcinoma and intraepithelial neoplasia].

OBJECTIVE: To investigate the expression and clinical significance of enhancer of zeste homolog 2 (EZH2) and p53 proteins in cervical squamous cell carcinoma (SCC) and cervical intraepithelial neoplasia (CIN). METHODS: The expression and distribution of EZH2 and p53 were determined with reference to clinicopathological features and patient survival. 168 cervical SCC, 19 CINII, 35 CINIII patients and 30 normal control cases were collected for immunohistochemical analysis. RESULTS: The expression of EZH2 in the normal cervix, CIN and SCC was 6.7% (2/30), 37.0% (20/54) and 75.6% (127/168), respectively, with a significant difference between them (P < 0.05). The expression of p53 was 3.3% (1/30), 20.4% (11/54) and 39.3% (66/168), respectively, also (P < 0.05). In the 168 SCC cases, the positive rate of EZH2 in the cases with lymph node metastasis was 82.9%, and that of p53 was 45.7%; the positive rate of EZH2/p53 protein expression in the cases with negative lymph nodes was 70.4%, and that of p53 was 34.7%, with a significant difference between the two subgroups (P < 0.05). Among the 143 followed-up SCC patients, the EZH2(+) p53(+) cases had a progression-free survival of (51.3 ± 3.8) months, that of EZH2(+) or p53(+) cases was (66.1 ± 2.0) months, and that of EZH2(-) p53(-) cases was (71.3 ± 1.9) months, with a very significant difference among the three subgroups (P < 0.001). The overall survival times of EZH2(-) p53(-), EZH2(+) or p53(+), and EZH(2+) p53(+) cases were (72.9 ± 1.1), (68.6 ± 1.8), and (57.4 ± 3.4) months, respectively, with a significant difference among the three subgroups (P < 0.001). The multivariate analysis showed that EZH2 expression, lymph node metastasis and tumor staging were independent prognostic factors. CONCLUSIONS: Both EZH2 and p53 proteins may play important roles in the occurrence and development of cervical squamous cell carcinoma. There is a close relationship between the expression of both EZH2 and p53 proteins and the prognosis of SCC patients.

LNMAC Promotes Cervical Squamous Cell Carcinoma Lymphatic Metastasis via Epigenetic Regulation of FGF2-Induced Lymphangiogenesis.

The lymph node is the most common site of distant metastasis of cervical squamous cell carcinoma (CSCC), which elicits dismal prognosis and limited efficiency for treatment. Elucidation of the mechanisms underlying CSCC lymphatic metastasis would provide potential therapeutic strategies for nodal metastatic of CSCC. Here, based on in vivo lymphatic metastasis screening model, a circular RNA is identified that is termed as lymph node metastasis associated circRNA (LNMAC), is markedly upregulated in lymphatic metastatic CSCC and correlated with lymph node metastasis. Overexpression of LNMAC dramatically augments the metastatic capability of CSCC cells to the lymph node via inducing lymphangiogenesis. Mechanistically, LNMAC epigenetically upregulates fibroblast growth factor 2 (FGF2) expression by directly associating with histone acacetylase 1 (HDAC1), preventing Importin α6/8-mediated nuclear translocation of HDAC1 and eliciting histone H3K27ac-induced FGF2 transcriptional activation. Treatment with 3F12E7, an anti-FGF2 monoclonal antibody, effectively inhibits LNMAC-induced CSCC lymphatic metastasis. Taken together, these findings indicate that LNMAC plays a crucial role in FGF2-mediated lymphangiogenesis and lymphatic metastasis, highlighting that LNMAC might be a therapeutic target for lymph node metastasis in CSCC patients.

Fatty Acid Oxidation Supports Lymph Node Metastasis of Cervical Cancer via Acetyl-CoA-Mediated Stemness.

Accumulating evidence indicates that metabolic reprogramming of cancer cells supports the energy and metabolic demands during tumor metastasis. However, the metabolic alterations underlying lymph node metastasis (LNM) of cervical cancer (CCa) have not been well recognized. In the present study, it is found that lymphatic metastatic CCa cells have reduced dependency on glucose and glycolysis but increased fatty acid oxidation (FAO). Inhibition of carnitine palmitoyl transferase 1A (CPT1A) significantly compromises palmitate-induced cell stemness. Mechanistically, FAO-derived acetyl-CoA enhances H3K27 acetylation (H3K27Ac) modification level in the promoter of stemness genes, increasing stemness and nodal metastasis in the lipid-rich nodal environment. Genetic and pharmacological loss of CPT1A function markedly suppresses the metastatic colonization of CCa cells in tumor-draining lymph nodes. Together, these findings propose an effective method of cancer therapy by targeting FAO in patients with CCa and lymph node metastasis.

Management and reproductive outcome of complete septate uterus with duplicated cervix and vaginal septum: review of 21 cases.

AIM: To evaluate the reproductive outcomes of complete septate uterus with duplicated cervix and vaginal septum. METHODS: Twenty-one patients who have complete septate uterus with duplicated cervix and vaginal septum were retrospectively reviewed in this study. In Group I, 11 patients with a poor reproductive outcome (spontaneous miscarriage or infertility) or dyspareunia underwent hysteroscopic metroplasty and removal of vaginal septum with the preservation of cervical septum. In Group II, 10 patients without a history of spontaneous miscarriage did not undergo hysteroscopic transection of the uterine septum. Of 10, four underwent vaginal septum incision due to dyspareunia, two underwent mere laparoscopic pelvic adhesiolysis because of infertility, and four without symptoms had no intervention. The primary endpoints included the pregnancy rate and outcomes of pregnancies. RESULTS: In Group I, the pregnancy rate after surgery is 81.8 % (9/11). Of nine women who conceived, six had term delivery, one encountered induced abortion due to the malformation of the fetal heart, and two had ongoing pregnancy. In Group II, among six patients accepting surgery, three had term delivery. The pregnancy rate after operation is 50 % (3/6). CONCLUSIONS: The uterine septum may not necessarily be transected for patients who have complete septate uterus with duplicated cervix and vaginal septum, and meanwhile have no a history of poor reproductive outcome.

Expression of human ß-defensin-2 in the eutopic and ectopic endometrial tissues in patients with endometriosis.

OBJECTIVE: To investigate the expression of human ß-defensin-2 (hBD-2) in the endometrium of patients with endometriosis (EMS) and explore the potential role of hBD-2 in the pathogenesis of EMS. DESIGN: Prospective controlled study. SAMPLE: 50 women including EMS patients undergoing laparoscopic ovarian cystectomy and non-EMS patients undergoing hysterectomy for uterine fibroids. SETTING: Large university teaching hospital. METHODS: Patients were divided into EMS and non-EMS groups. The gene expressions of hBD-2, interleukin (IL)-1ß and tumor necrosis factor (TNF)-α in the endometrial tissues of each group were detected with real-time quantitative polymerase chain reaction (PCR), and hBD-2 protein expression with immunohistochemical method. RESULTS: The gene expression levels of hBD-2, TNF-α, and IL-1ß as well as the positive expression rate of hBD-2 protein in the ectopic endometrium of EMS patients were significantly higher than those in the eutopic endometrium of EMS and non-EMS patients (all P < 0.05). Correlation analysis showed that the gene expression levels of hBD-2 in the ectopic and eutopic endometrium of EMS patients were positively correlated with the gene expression levels of IL-1ß and TNF-α (P < 0.01). CONCLUSION: High levels of hBD-2 gene and protein expressions in the ectopic endometrium of EMS patients may be an important contributor in the pathogenesis of EMS. TNF-α and IL-1ß may promote the upregulation of hBD-2 expression.

lncRNA GAS5 suppression of the malignant phenotype of ovarian cancer via the miR-23a-WT1 axis.

Background: Growth arrest-specific 5 (GAS5) is a long noncoding RNA (lncRNA) that regulates cell viability. GAS5 lncRNA has been shown to decrease colorectal and breast cancer carcinogenesis. Although the function and mechanisms related to lncRNA GAS5 in the development of ovarian cancer (OC) remains unclear. The goal of this study was to clarify the essential functions of lncRNA GAS5 in regulating OC progression and its underlying mechanism. Methods: Relative levels of lncRNA GAS5 in OC and normal ovarian tissues were identified by quantitative real-time polymerase chain reaction (qRT-PCR). The regulatory effects of lncRNA GAS5 on the proliferation and apoptosis of SK-OV-3 cells were evaluated. Moreover, bioinformatics tools were used to predict a novel target [microRNA (miRNA)] of lncRNA GAS5. To explore the key functions of the lncRNA GAS5/miRNA-23a/WT1 regulatory loop in mediating OC progression, we performed functional experiments and a dual-luciferase reporter (DLR) gene assessment. Results: Downregulation of lncRNA GAS5 was found in tissues of OC, which was positively correlated with a poor prognosis. In addition, SK-OV-3 cells with a lower expression of lncRNA GAS5 and accelerated cancer cell migration demonstrated a lower percentage of apoptosis in in vitro experiments. It was demonstrated that lncRNA GAS5 acts as a molecular sponge for miR-23a in OC cells. Additionally, WT1 was detected as a miR-23a target gene in OC cells, and through sponging miR-23a, lncRNA GAS5 positively regulated WT1 expression. Rescue tests demonstrated that enhancing the outputs of the miR-23a-WT1 axis reversed the impacts of lncRNA GAS5 silencing on cell proliferation and apoptosis in OC. Conclusions: The lncRNA GAS5/miR-23a/WT1 cascade was found participate in the progression of OC. lncRNA GAS5 also decreases OC progression by upregulating WT1 and attenuating miR-23a, suggesting that it could be an advantageous therapeutic target for OC intervention.

Effects of leisure activities and general health on the survival of older people: a cohort study in China.

Objective: This study aimed to examine the influence of physical and cognitive leisure activities on the survival of older adults in China, while also exploring the potential mediating and moderating effects of general health. Methods: This study utilized the China Longitudinal Healthy Longevity Survey (CLHLS) datasets spanning from 2008 to 2018, and 10,347 eligible participants were included. The primary study outcome was all-cause mortality, and independent variables included physical leisure activities (PLA), cognitive leisure activities (CLA), and self-rated general health. Three sets of covariates were adjusted, including socio-demographic characteristics, health behaviors, and health status. Results: The longest survival time was the older people participating in PLA & CLA (mean = 50.31 months), while those participating in neither exhibited the lowest (mean = 29.60 months). Significant differences in survival status were observed in different types of leisure activities participation (Log-rank test, Chi-square = 576.80, p < 0.001). Cox regression indicated that PLA (HR = 0.705, 95% CI: 0.651-0.764), CLA (HR = 0.872, 95% CI: 0.816-0.933), and the both PLA & CLA (HR = 0.700, 95% CI: 0.656-0.747) were protective factors for the survival. Additionally, general health significantly moderated the relationship between PLA and reduced mortality risk (Coefficient = -0.089, p = 0.042). While CLA indirectly influenced the survival through general health (Coefficient = -0.023, p < 0.001). For the older people participating in PLA and CLA, general health played mediating (Coefficient = -0.031, p < 0.001) and moderating (Coefficient = -0.026, p = 0.013) role in the relationship between leisure activities and survival. Conclusion: Leisure activities and self-rated general health were important predictors of survival of the older adults, and general health exhibited a mediator and moderator in the relationship between leisure activities and survival status.

Impact of lifestyle and psychological resilience on survival among the oldest-old in China: a cohort study.

Introduction: Healthy lifestyles and psychological resilience are important factors influencing the life expectancy of the oldest-old (≥80 years). Stratified by urban and rural groups, this study used a 10-year cohort to examine the mechanism of lifestyle and psychological resilience on the survival of the oldest-old in China. Methods: This study used the China Longitudinal Healthy Longevity Survey datasets spanning from 2008 to 2018, and 9,250 eligible participants were included. The primary outcome variable was all-cause mortality, and independent variables included healthy lifestyle index and psychological resilience. Six covariates were included in the survival analysis and moderation-mediation model, such as gender and annual household income. Results: This study found that the oldest-old with five healthy lifestyles had the longest survival time, averaging 59.40 months for urban individuals and 50.08 months for rural individuals. As the lifestyle index increased, the survival rate significantly increased. The Cox regression showed that for the urban oldest-old, the lifestyle index served as a protective factor for survival outcomes. However, this effect lost statistical significance among rural oldest-old individuals. For urban oldest-old individuals, psychological resilience significantly mediated and moderated the effect of the lifestyle index on survival status, but the moderating effect was not statistically significant for the rural ones. Discussion: Overall, healthy lifestyles and psychological resilience can be effective in enhancing the survival of the oldest-old, and there are differences between urban and rural population, so different interventions should be adopted for urban and rural areas to achieve longer life in China.

Fucosterol exhibits selective antitumor anticancer activity against HeLa human cervical cell line by inducing mitochondrial mediated apoptosis, cell cycle migration inhibition and downregulation of m-TOR/PI3K/Akt signalling pathway.

[This retracts the article DOI: 10.3892/ol.2018.7769.].