LncRNA NEAT1 inhibits apoptosis and autophagy of ovarian granulosa cells through miR-654/STC2-mediated MAPK signaling pathway.

Long non-coding RNA (lncRNA) anomalies cause early ovarian failure. LncRNA nuclear enriched abundant transcript 1 (NEAT1) was down-regulated in premature ovarian failure (POF) mice and connected to the illness, however, the mechanism remained unclear. The levels of gene and protein were measured by using quantitative real-time polymerase chain reaction, Western blot, and immunofluorescence. Follicle stimulating hormone (FSH), estradiol (E2), and luteinizing hormone (LH) levels were determined using enzyme-linked immunosorbent assay (ELISA). 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) and flow cytometry were used to determine cell viability and apoptosis. The interaction of NEAT1, miR-654, and stanniocalcin-2 (STC2) was verified by dual-luciferase reporter assay or RNA binding protein immunoprecipitation (RIP) assays. The results showed NEAT1 and STC2 down-regulated, while miR-654 up-regulated in POF mice. Overexpression of NEAT1 reduced apoptosis and autophagy in cyclophosphamide (CTX)-treated ovarian granulosa cells (OGCs), and Bax, cleaved-caspase3, LC3B, LC3II/LC3I ratio were decreased and Bcl-2 and p62 were raised. NEAT1 suppressed miR-654 expression by directly targeting miR-654. The inhibition of NEAT1 overexpression on apoptosis and autophagy in OGCs was reversed by miR-654 mimics. STC2 was a target gene of miR-654, and miR-654 inhibitor reduced the apoptosis and autophagy by regulating the STC2/MAPK axis. To sum up, NEAT1 reduced miR-654 expression and modulated the STC2/MAPK pathway to decrease apoptosis and autophagy in POF, indicating a potential therapeutic target.

Seasonal Disparities of Human Health Risk and Particle-Bound Metal Characteristics Associated with Atmospheric Particles in a Fishery Harbor.

The effects of atmospheric pollution from ship emissions have been considered for several harbors worldwide. The health risk assessment and source apportionment of particle-bound metals in a fishery harbor were investigated in this study. The most abundant metal elements in particulate matter (PM) on all sampling days in three seasons were Fe (280.94 ± 136.93 ng/m3), Al (116.40 ± 71.25 ng/m3), and Zn (110.55 ± 26.70 ng/m3). The ratios of V/Ni were 1.44 ± 0.31, 1.48 ± 0.09 and 1.87 ± 0.06 in PM10, PM2.5, and PM1, respectively. Meanwhile, the ratios higher than 1 indicated that fuel oil combustion from ship emission in fishery harbor. The highest deposits of total particle-bound metals in the human respiratory tract were in the head airway (HA), accounting for 76.77 ± 2.29% of the total particle-bound metal concentration, followed by 5.32 ± 0.13% and 2.53 ± 0.15% in the alveolar region (AR) and tracheobronchial (TB) region, respectively. The total cancer risk (CR) of inhalation exposure to local residents exceeded 10-6. Mean total CR values followed the sequence: autumn (1.24 × 10-4) > winter (8.53 × 10-5) > spring (2.77 × 10-6). Source apportionment of related metal emissions was mobile pollution emissions (vehicle/boat) (37.10-48.92%), metal fumes of arc welding exhaust (19.68-34.42%), spray-painting process (12.34-16.24%), combustion emissions (6.32-13.12%), and metal machining processes (9.04-16.31%) in Singda fishing harbor. These results suggest that proper control of heavy metals from each potential source in fishing harbor areas should be carried out to reduce the carcinogenic risk of adverse health effects.

Assessment of the Association between Genetic Polymorphisms in the CHI3L1 Gene and Asthma Risk.

BACKGROUND: Chitinase 3-like 1 (CHI3L1) is an important factor involved in the development of asthma. This meta-analysis assessed the association of the CHI3L1 polymorphisms rs4950928, rs10399931, rs883125, rs880633, and rs10399805 with asthma risk. METHODS: The literature searches were conducted in PubMed, Web of Science, Wanfang, and China National Knowledge Infrastructure up until September 4, 2021, for relevant studies. Sixteen publications with 18 studies involving 5,005 asthma patients and 9,725 controls were included in this meta-analysis. RESULTS: The meta-analyses showed that among East-Asian subjects, increased asthma risk was associated with CHI3L1 rs4950928 (GG + CG vs. CC: odds ratio [OR] = 1.43, 95% confidence interval [CI]: 1.09-1.88, p = 0.011; GG vs. CG + CC: OR = 1.64, 95% CI: 1.20-2.26, p = 0.002; GG vs. CC: OR = 1.97, 95% CI: 1.41-2.75, p = 0.000; and G vs. C: OR = 1.36, 95% CI: 1.12-1.66, p = 0.002) and rs883125 (G vs. C: OR = 1.42, 95% CI: 1.01-1.99, p = 0.043), whereas CHI3L1 rs10399931 was associated with reduced asthma risk (TT vs. CT + CC: OR = 0.79, 95% CI: 0.64-0.99, p = 0.038; TT vs. CC: OR = 0.77, 95% CI: 0.61-0.98, p = 0.030). In addition, we found an association between CHI3L1 rs4950928 and asthma risk in adult subjects but not children, while CHI3L1 rs883125 was associated with asthma risk in children. CONCLUSION: The CHI3L1 polymorphisms rs4950928, rs10399931, and rs883125 are important genetic factors for asthma among East-Asian subjects.

Dietary Fiber and Human Papillomavirus Infection among US Women: The National Health and Nutrition Examination Survey, 2003-2016.

The impact of dietary fiber on human papillomavirus (HPV) infection is still underway. The aim of our study was to investigate the association between intake of dietary fiber and HPV infection. Overall, 14,151 eligible women, aged 18-59 years old, who submitted an adequate sample for HPV test, were collected from an ongoing, large scale population-based survey for seven cycles. The association of dietary fiber intake and HPV infection was assessed in multivariate logistic models. For sensitivity analysis, generalized additive model (GAM) and smooth curve fitting were employed to verify the robustness of the results. Among 14,151 eligible participants, intake of dietary fiber was negatively associated with HPV infection. Each additional increase in log10 dietary fiber consumption was associated with a 57% lower risk of HPV infection (OR, 0.43; 95% CI 0.38-0.48). The result is stable in minimally and fully adjusted model. The possibility of nonlinear association of dietary fiber and HPV infection has been excluded by GAM and smooth curve fitting. There was an inverse linear correlation between intake of dietary fiber and HPV infection. Our findings obtained from NHANES dataset suggested that increasing dietary fiber consumption may be associated with the prevalence of HPV infection.

The women's heart health programme: a pilot trial of sex-specific cardiovascular management.

BACKGROUND: There is increasing knowledge of sex-specific differences in cardiovascular disease and recognition of sex disparities in management. In our study, we investigated whether a cardiovascular programme tailored to the specific needs of women could lead to improved outcomes. METHODS: We randomised 100 female patients to receive cardiology follow-up with the conventional sex-neutral cardiac programme (control), or the sex-tailored Women's Heart Health Programme (intervention). The intervention group was managed by an all-women multidisciplinary team and received culture-centred health intervention workshops, designed through in-depth interviews with the participants. The primary outcome was cardiovascular risk factor improvement at 1 year. Secondary outcomes include cardiovascular event rates, quality of life scores, and self-reported improvement in knowledge, attitudes, intentions and practices. Generalised structural equation model analysis was used to determine if the intervention group had better outcomes at alpha level 0.1. RESULTS: The mean age was 67.3 ± 12.7 years, with an ethnic distribution of 70% Chinese, 18% Malays, and 12% Indians. The majority of these patients had no formal or primary level of education (63%), and were mostly unemployed (78%). Patients in intervention group had better control of diabetes mellitus (lower HbA1c of 0.63% [CI 0.21-1.04], p = 0.015) and lower body-mass-index (0.74 kg/m2 [CI 0.02-1.46], p = 0.092) at 1 year, but there was no significant difference in blood pressure or lipid control. Overall, there was a trend towards better risk factor control, 31.6% of intervention group versus 26.5% of control group achieved improvement in at least 1 CV risk factor control to target range. There was no significant difference in incidence of cardiovascular events, quality of life, or domains in knowledge, attitudes, intention and practices. CONCLUSION: This pilot study is the first of its kind evaluating a new model of care for women with heart disease. The potential to improve outcomes needs to be studied in a larger trial with longer follow up. TRIAL REGISTRATION: This trial was prospectively registered clinicaltrials.gov on 6 May 2013. Trial Number: 2013/00088. Identifier: NCT02017470.

Single nucleotide polymorphisms of IL-13 and CD14 genes in allergic rhinitis: a meta-analysis.

PURPOSE: The aim of this study was to conduct a meta-analysis for single nucleotide polymorphisms (SNPs) in interleukin-13 (IL-13) and cluster of differentiation 14 (CD14) genes and the risk for allergic rhinitis (AR). METHODS: We screened studies identified through seven databases including Pubmed, Medline, Web of Science, Embase, China Biology Medicine disc, Wanfang, and China Academic Journal Network Publishing Database. The odds ratios (ORs) and 95% confidence intervals (CIs) were determined to assess the association under allelic, dominant and recessive models. RESULTS: Twelve studies with a total of 8547 participants (3223 cases and 5324 controls) investigated IL-13 SNP rs20541, five studies combining 4580 participants (1411 cases and 3169 controls) examined IL-13 SNP rs1800925, and nine studies with 2301 participants (1174 cases and 1127 controls) assessed CD14 SNP rs2569190. We found that the A allele of IL-13 SNP rs20541 was associated with an increased risk of AR (OR 1.19, 95% CI 1.11-1.28, P < 0.001). Stratifying studies by ethnic group produced significant results in Asians (OR 1.21, 95% CI 1.11-1.32, P < 0.001), but not in Caucasians (OR 1.14, 95% CI 1.00-1.30, P = 0.051). No association of IL-13 SNP rs1800925 and CD14 SNP rs2569190 with AR risk was found in either Asians or Caucasians (P > 0.05). CONCLUSION: Our findings suggest that IL-13 SNP rs20541 is significantly associated with AR risk in Asians but not in Caucasians. However, the accumulating evidence does not support an association of IL-13 SNP rs1800925 and CD14 SNP rs2569190 with AR risk.

Lack of Association Between the IL1B (-511 and +3954), IL1RN VNTR Polymorphisms and Tuberculosis Risk: A Meta-analysis.

BACKGROUND: Several recent studies have provided evidence that polymorphisms in the interleukin-1 (IL1) gene are implicated in tuberculosis (TB). However, results of different studies are inconsistent. The aim of this study was to perform a meta-analysis investigating the association of the IL1B (-511 and +3954) and IL1RN VNTR polymorphisms with TB risk. METHODS: A systematic review of the English literature was conducted by searching Pubmed, Scopus, and ISI Web of Knowledge databases for relevant studies. Pooled odds ratios (OR) with 95 % confidence intervals (CI) were calculated using fixed effects models. Between-study heterogeneity and publication bias were also evaluated. RESULTS: Nine case-control studies including 3327 participants were reviewed and analyzed. Our results did not indicate any association of the IL1B (-511 and +3954) and IL1RN VNTR polymorphisms with TB risk in the overall populations. The pooled OR of the IL1B -511 polymorphism was 1.09 (95 % CI 0.87-1.36) for the dominant model, 1.11 (0.89-1.38) for the recessive model, 1.15 (0.87-1.50) for the homozygote model, and 1.07 (0.94-1.23) for the allelic comparison model. ORs for the IL1B +3954 and IL1RN VNTR polymorphisms were similar. In subgroup analysis stratified by ethnicity, the results revealed no association between these polymorphisms and TB risk in black people, Asians, and Caucasians, respectively. We did not identify significant between-study heterogeneity across all studies, and there was no evidence of publication bias. CONCLUSIONS: Our results indicate there is a lack of association between the IL1B (-511 and +3954), IL1RN VNTR polymorphisms and TB risk.

[Influencing factors for severity of bronchopulmonary dysplasia in preterm infants].

OBJECTIVE: To explore the influencing factors for the severity of bronchopulmonary dysplasia (BPD) in preterm infants. METHODS: The clinical data of 110 preterm infants who were diagnosed with BPD and had a hospital stay of over 28 days between January 2011 and December 2013 were analyzed. These BPD infants were divided into 3 groups according to the clinical criteria: mild group (n=52), moderate group (n=44), and severe group (n=14). The relationship between the severity of BPD and the gestational age, birth weight, asphyxia, oxygen therapy, pregnancy complications, intrauterine pneumonia and mechanical ventilation was analyzed. RESULTS: The severity of BPD was correlated with the following factors: gestational age, birth weight, prenatal infection, duration of oxygen inhalation with a concentration of >40%, use of mechanical ventilation, parameters and duration of mechanical ventilation, duration of continuous positive airway pressure, adoption of intubation surfactant extubation (INSURE) approach, Ureaplasma urealyticum infection, intrauterine pneumonia and patent ductus arteriosus. Logistic regression analysis indicated that the mechanical ventilator parameter peak inspiratory pressure (OR=1.260, 95%CI: 1.096-1.448) and duration of mechanical ventilation (OR=1.010, 95%CI: 1.005-1.016) were independent risk factors for the severity of BPD, while the INSURE approach was a protective factor (OR=0.208, 95%CI: 0.060-0.923). CONCLUSIONS: The severity of BPD is associated with various factors in preterm infants. The important measures for preventing BPD include avoiding the birth of preterm infants with a very low birth weight, shortening the duration of mechanical ventilation, preventing and reducing pulmonary infections, and applying the INSURE approach.

Shikonin from lithospermum erythrorhizon induces pyroptosis in trophoblast cells by activating the CTSB-NLRP3 inflammasome.

BACKGROUND: With the decline of global fertility, drug therapeutic of ectopic pregnancy is of great significance. Lithospermum erythrorhizon is using for embryo killing as herbal medicine. Shikonin is the critical nucleus of Lithospermum erythrorhizon; however, the mechanism is still unclear. The study aimed to explore the mechanism of shikonin against ectopic pregnancy. MATERIAL AND METHODS: In this study, we examined the viability and LDH release of HTR-8/SVneo cells by assays, observed pore formation in cell membranes by microscopy imaging and PI staining, and IL-1ß release by WB and ELISA assay kit. Then, we used network pharmacology to analyse the potential interaction between shikonin, ectopic pregnancy and pyroptosis and used molecular docking techniques to verify interactions between shikonin and core common targets. Finally, western blotting and immunofluorescence assay were used to explore the mechanism of shikonin-inducing pyroptosis of HTR-8/SVneo cells. RESULTS: Shikonin could cause a significant inhibition of HTR-8/SVneo cell viability in a concentration- and time-dependent manner. In HTR-8/SVneo cells, shikonin-induced cell swelling, bubble formation, an increase in the release of lactate dehydrogenase (LDH) and up-regulation of several pyroptosis-associated factors. And network pharmacology showed that The main targets of shikonin-ectopic pregnancy-pyroptosis were IL-1ß and caspase-1, and molecular docking results showed that shikonin can closely bind to IL-1ß, caspase-1 and GSDMD. Additionally, the necroptosis inhibitor GSK'872 could not suppress the expression of mature-IL-1ß and prevent the pyroptosis phenotype from developing. However, the nucleotide oligomerization domain-like receptor family pyrin domain containing 3 (NLRP3) inhibitor MCC-950 could downregulate the expression of pyroptosis-associated factors and prevent the pyroptosis phenotype from developing. Shikonin led to an elevation in the expression of cathepsin B (CTSB), and the CTSB inhibitor CA-074 abolished pyroptosis induced by shikonin; however, the NLRP3 inhibitor MCC-950 could not inhibit the expression of CTSB. CONCLUSIONS: Our results suggest that shikonin activates CTSB to induce NLRP3-dependent pyroptosis in HTR-8/SVneo cells. This study has important clinical implications for the treatment of ectopic pregnancy.

Association of the MMP-3, MMP-9 and MMP-12 gene polymorphisms with COPD risk: a meta-analysis.

Introduction: Given the evidence that the matrix metalloproteinases (MMPs) play an important role in the pathogenesis of chronic obstructive pulmonary disease (COPD), a number of case-control studies have attempted to assess the relationship between genetic polymorphisms in MMP genes and COPD risk. However, reliable measures of these results are lacking. Material and methods: We assessed the published evidence for association of the MMP-3, MMP-9 and MMP-12 polymorphisms with COPD risk using meta-analytic techniques. The odds ratio (OR) and 95% confidence interval (CI) were calculated for each study using fixed or random effect models. Results: A total of 23 case-control studies were included in the meta-analysis. No significant association was observed between the MMP-9 rs3918242 polymorphism and COPD risk in the overall populations under the dominant (T/T + C/T vs. C/C: OR = 1.30, 95% CI: 1.00-1.69, p = 0.054) and allele contrast (T allele vs. C allele: OR = 1.22, 95% CI: 0.97-1.53, p = 0.088) models. However, in sub-group analysis the polymorphism rs3918242 was significant in Asians under the dominant model (T/T + C/T vs. C/C: OR = 1.66, 95% CI: 1.02-2.72, p = 0.043). The results for MMP-12 rs2276109 showed an association with COPD only in mixed populations (G/G + A/G vs. A/A: OR = 1.57, 95% CI: 1.10-2.24, p = 0.013; G allele vs. A allele: OR = 1.52, 95% CI: 1.09-2.14, p = 0.015). We did not find any significant association of the MMP-12 rs652438 and MMP-3 rs35068180 polymorphisms with COPD. Conclusions: The findings of this meta-analysis suggest that there is a risk of COPD associated with the MMP-9 rs3918242 and MMP-12 rs2276109 polymorphisms in certain ethnic groups.

[Changes of CD(4)(+)Foxp3(+) regulatory T cells and CD(4)(+)IL-17(+)T cells in cigarette smoke-exposed rats].

OBJECTIVE: To evaluate the changes of CD(4)(+)IL-17(+) T (Th17) and CD(4)(+)Foxp3(+) regulatory T (Treg) cells in peripheral blood and bronchoalveolar lavage fluid (BALF), and therefore to explore the role of Th17 and Treg in cigarette smoke-induced airway inflammation/COPD in rats. METHODS: Forty male Wistar rats were randomly divided into 4 groups: a 12 wk smoke-exposure group, a 24 wk smoke-exposure group, a 12 wk control group and a 24 wk control group (n = 10 each). Cells in BALF were collected and analyzed by absolute and differential cell counts. IL-17 and IL-6 levels in serum and BALF were tested by enzyme linked immunosorbent assay (ELISA). The proportion of CD(4)(+)IL-17(+) T and CD(4)(+)Foxp3(+) Treg in peripheral blood and BALF were determined by flow cytometry. The mRNA expressions of IL-17 and Foxp3 were measured by real-time PCR. Comparisons of the data between different groups were performed using one-way ANOVA, and SNK and Games-Howell test were used for comparison between 2 groups. RESULTS: Levels of IL-17 were remarkable increased in the 12 wk smoke-exposure group and the 24 wk smoke-exposure group in serum [(52.6 ± 1.8) ng/L, (75.4 ± 6.0) ng/L] and BALF [(78.1 ± 5.8) ng/L, (95.0 ± 6.8) ng/L] compared with the 12 wk control group [(40.0 ± 3.2)ng/L, (54.5 ± 4.6) ng/L] and the 24 wk control group [(36.7 ± 3.2) ng/L, (53.9 ± 3.7) ng/L], all P < 0.05. IL-6 in serum was significantly increased in the 24 wk smoke-exposure group [(31.4 ± 2.1) ng/L] compared with the 24 wk control group [(11.5 ± 0.5) ng/L], and it was increased in the 12 wk and the 24 wk smoke-exposure group [(33.3 ± 2.3) ng/L, (44.6 ± 3.0) ng/L] compared with the 12 wk and the 24 wk control group [(15.6 ± 1.8) ng/L, (18.0 ± 1.9) ng/L] in BALF. Ratio of Th17 was higher in the 12 wk and the 24 wk smoke-exposure groups in peripheral blood [(1.81 ± 0.19)%, (3.74 ± 0.55)%] and BALF [(7.84 ± 0.28)%, (8.01 ± 0.39)%] compared with the12 wk [(0.97 ± 0.08)%, (5.64 ± 0.54)%] and the 24 wk control group [(1.08 ± 0.10)%, (5.95 ± 0.48)%]. Ratio of Treg in BALF was higher in the smoke-exposure groups [(8.81 ± 0.49)%, (11.98 ± 0.72)%] compared with the control groups [(4.34 ± 0.28)%, (5.21 ± 0.42)%]. The level of IL-17 mRNA was increased in the 12 wk and the 24 wk smoke-exposure group in peripheral blood (25.7 ± 2.0, 33.9 ± 1.5) and in BALF (22.2 ± 1.8, 34.7 ± 4.2) compared with the 12 wk (11.3 ± 2.6, 11.6 ± 2.4) and the 24 wk (11.1 ± 2.0, 13.5 ± 3.4) control groups. Foxp3 mRNA was increased in the smoke-exposure groups (24.4 ± 2.7, 30.3 ± 2.7) compared with the control groups (12.7 ± 2.7, 14.6 ± 3.8). Th17 in smoke-exposure groups was positively correlated with counts of total cells and macrophages (r = 0.512, 0.543, all P < 0.05). CONCLUSIONS: An elevated expression of Th17 and Treg cells and an increase of inflammatory cytokines were evident in airway inflammation of cigarette smoke-exposed rats, suggesting that Treg was involved in the immunological regulation and Th17 was associated with the persistent inflammation in cigarette smoke-induced airway inflammation in rats.

Research status and hotspots of cesarean scar pregnancy from 2001 to 2020: a bibliometric analysis.

BACKGROUND: Due to the significant increase in cesarean section rates worldwide, cesarean scar pregnancy (CSP) has attracted extensive attention. This study aimed to investigate the global research status and developmental directions of CSP to discern promising research hotspots by means of bibliometrics and visualized analyses. METHODS: The English CSP-related literature from 2001 to 2020 was retrieved from The Web of Science Core Collection (WOSCC) database. Microsoft Excel 2019 was used to analyze the years, countries, institutions, journals, authors, citations, H indices, titles, abstracts, and keywords of related literature, and VOSviewer software was used to visualize developmental directions and promising hotspots in CSP. RESULTS: The study included 1,186 articles in total. The number of CSP articles has presented an overall increasing trend over the last two decades. China has the maximum number of publications, but America's H-index is higher than that of China, with the average number of citations per item ranking fourth. The Journal of Obstetrics and Gynaecology Research published the most on the subject, while Zhejiang University from China published the largest number of articles; Jurkovic D, Timor-Tritsch IE, and Monteagudo A have achieved considerable progress in the CSP domain. There are 5 clusters that the current research orientation can fall into: "selection of the delivery mode with a scarred uterus after cesarean section", "risk factors", "diagnosis", "treatment" and "related basic and clinical experiments". High-intensity focused ultrasound (HIFU) and the formulation of consensus guidelines are the focus of the current research. CONCLUSIONS: The analysis of global trends shows that CSP research is being actively studied and that the number of published studies is continuously increasing. According to the comprehensive quality and quantity of the literature, the United States maintains a leading position in CSP research. In recent years, the research directions have mainly focused on "diagnosis", "treatment" and "risk factors", while HIFU and the formulation of consensus guidelines may become research hotspots in the field of CSP.

Shikonin Could Be Used to Treat Tubal Pregnancy via Enhancing Ferroptosis Sensitivity.

Background: Albeit oxidative stress has been implied in the pathogenesis of tubal pregnancy (TP), there are scant data to suggest that ferroptosis occurs in TP. Shikonin plays a pivotal role in redox status, but whether it can regulate ferroptosis to treat TP remains unknown. Methods: We collected and analyzed ferroptosis-related indices from the villous tissue (VT) of women suffering from TP and from women with a normal pregnancy. In vitro, we used shikonin and/or RAS-selective lethal 3 (RSL3) to intervene HTR-8/SVneo cells and further detected ferroptosis indices and cell functions. Finally, the expression of the nuclear factor erythroid 2-related factor 2 (Nrf2) is pharmacologically activated to explore the effect of Nrf2 on shikonin regulating ferroptosis. Results: Increased malondialdehyde content, reduced levels of glutathione and glutathione peroxidase (GPx), and upregulated protein expression which promoted ferroptosis were observed in the VT of TP patients, suggesting that ferroptosis occurred during TP. In vitro, shikonin enhanced ferroptosis sensitivity in HTR-8/SVneo cells induced by RSL3 via amplifying lipid peroxidation, which mainly included increasing cellular reactive oxygen species (ROS), lipid ROS and Fe2+ level. RSL3 and/or shikonin inhibited Nrf2 and downregulated protein expression of SLC7A11 and GPx4 caused by RSL3 + shikonin co-treatment, which could be reversed under activation of Nrf2. Hence, shikonin facilitated lipid peroxidation by inhibiting Nrf2 signaling. Additionally, shikonin and/or RSL3 potently inhibited the invasion and migration of HTR-8/SVneo cells. Conclusion: This study firstly showed that ferroptosis may be involved in TP pathogenesis and shikonin potentially targeted ferroptosis to treat TP.

LncRNA GATA3-AS1 promoted invasion and migration in human endometrial carcinoma by regulating the miR-361/ARRB2 axis.

Endometrial carcinoma (EC) is a kind of fatal female malignancy. lncRNA GATA3-AS1 has been identified as an oncogene in various cancers. However, the functions and mechanisms of GATA3-AS1 in EC remain to be explored. Human EC tissues and four EC cell lines were used. Western blotting and quantitative real-time PCR (qRT-PCR) were used to evaluate the expression of GATA3-AS1, miR-361, and ARRB2. Dual-luciferase reporter and RNA immunoprecipitation (RIP) assays were used to validate the interaction among GATA3-AS1, miR-361, and ARRB2. Flow cytometry, colony formation assay, scratch assay, and transwell assay were used to examine the cell apoptosis, proliferation, migration, and invasion of EC cells, respectively. In vivo tumor growth was monitored in nude mice. GATA3-AS1 and ARRB2 were upregulated while miR-361 was downregulated in human EC tissues and EC cells. GATA3-AS1 knockdown constrained cell proliferation, invasion, migration, and EMT while promoting the apoptosis of EC cells by upregulating miR-361. GATA3-AS1 negatively regulated miR-361 expression. ARRB2 was the direct target of miR-361 and could activate the Src/Akt pathway. In vivo, GATA3-AS1 knockdown suppressed tumor progression by upregulating the miR-361 expression. lncRNA GATA3-AS1 promoted EC invasion and migration by the miR-361/ARRB2 axis, which indicated that GATA3-AS1 might be a promising therapeutic option for advanced EC progression. KEY MESSAGES: GATA3-AS1 knockdown suppressed EC proliferation, invasion, and migration. GATA3-AS1 directly inhibited miR-361 as a ceRNA. MiR-361 knockdown reversed the tumor suppressive effect caused by GATA3-AS1 knockdown. MiR-361 bound to ARRB2 directly and suppressed its expression. The GATA3-AS1/miR-361/ARRB2 axis regulated EC cell proliferation, invasion, and migration.

Association Between White Blood Cells at Baseline and Treatment Failure of MTX for Ectopic Pregnancy.

Purpose: The aim of this study was to evaluate white blood cell (WBC) count as a risk factor related to methotrexate (MTX) treatment failure in patients with ectopic pregnancy (EP). Methods: A total of 236 women diagnosed with EP and treated with a single dose of MTX were included. The exposure variable was WBC count at baseline, and the outcome was MTX treatment outcome. Both a multivariate binary logistics regression model and subgroup analysis were performed to evaluate the association between WBC and MTX non-response. Results: WBC count was associated with the risk of treatment failure, and the odds ratio (OR) in different multivariate models was stable [minimally adjusted model: OR 1.2, 95% confidence interval (CI): 1.0-1.3, p = 0.008; fully adjusted model: OR 1.2, 95% CI: 1.0-1.4, p = 0.026]. For WBCs in group T3 (>8.9 × 109/L), the association between WBC count and treatment failure was significant (minimally adjusted model: OR: 2.0, 95% CI: 1.0-3.8, p = 0.050; fully adjusted model: OR: 2.2, 95% CI: 1.1-5.6, p = 0.034). Subgroup analysis showed that in participants with regular menstruation (OR 1.1, 95% CI: 1.0-1.3), WBC count was significantly different from irregular menstruation (OR 1.8, 95% CI: 1.2-2.8); p for interaction was 0.031. Conclusions: We found a reliable and non-linear relationship between WBC count and MTX treatment failure for EP.

[Change of airway inflammation induced by Th1/Tc1 and the expression of T regulatory cells in smoking cessation rats].

OBJECTIVE: to study the change of airway inflammation induced by Th1/Tc1 and the expression of CD4(+)CD25(+) regulatory T cells (Treg) in smoking cessation rats. METHODS: fifty healthy male Wistar rats were randomly divided into five groups: 12-week normal control (group A, n = 10), 24-week normal control (group B, n = 10), 12-week smoke exposure (group C, n = 10), 24-week smoke exposure (group D, n = 10) and smoking cessation (group E, n = 10). Groups C, D and E were exposed to cigarettes for 12 weeks. At Week 12, groups A and C were sacrificed. Group D continued smoke exposure and group E had smoking cessation for 12 weeks. At Week 24, groups B, D and E were sacrificed. Pathomorphological changes of small airway were analyzed. The cells in BALF (bronchoalveolar lavage fluid) were collected and analyzed by absolute and differential cell counts. Enzyme linked immunosorbent assay (ELISA) was used to detect the levels of IFN-γ, IL-4, IL-8 and TNF-α. And flow cytometry was employed to determine the Foxp3 + Treg cell populations and reverse transcription-polymerase chain reaction (RT-PCR) to assay the mRNA expression for Foxp3. RESULTS: (1) compared with groups A and B, the airway inflammation score of groups C, D and E increased significantly (all P < 0.01). Compared with group C, the airway inflammation score of groups D and E both increased (all P < 0.01), especially group D; (2) compared with groups A and B, the levels of IFN-γ, TNF-α and IL-8 in groups C, D and E increased (all P < 0.01) while those of IL-4 decreased. The levels of IFN-γ, TNF-α and IL-8 showed no difference between groups C and E. The levels of IFN-γ, TNF-α and IL-8 were higher in group D than those in groups C and E; (3) the ratio of Foxp3 + Treg cells in BALF was higher in group C (7.4% ± 0.8%), group D (7.8% ± 1.7%) and group E (7.0% ± 1.4%) than group A (4.8% ± 1.2%) and group B (4.7% ± 1.2%) (all P < 0.01). There were no differences in the ratio of Foxp3 + Treg cells among groups C, D and E (all P < 0.05); (4) there was an elevated expression of Foxp3 mRNA in group C (0.22 ± 0.02), group D (0.23 ± 0.03), group E (0.20 ± 0.04) versus group A (0.13 ± 0.01) and group B (0.11 ± 0.02) (all P < 0.01). But there was no difference in the expression of Foxp3 mRNA among groups C, D and E (all P > 0.05). CONCLUSIONS: airway inflammation induced by Th1/Tc1 and an elevated expression of Treg cells in BALF are found in smoke exposure rats. Upon smoking cessation, the above-mentioned airway inflammation still persists and the expression of Treg cells in BALF shows no decrease. It suggests that an immune imbalance may be involved in the progression of Th1/Tc1-induced airway inflammation upon smoking cessation.

[CD4(+)Foxp3(+) regulatory T cells in inflammation and emphysema after smoking cessation in rats].

OBJECTIVE: To evaluate the expression of Treg in a cigarette smoke-induced rat model of emphysema and after smoking cessation in the rats. METHODS: Fifty male Wistar rats were randomly divided into control group 1 (12 weeks), control group 2 (24 weeks), smoke-exposure group 1 (12 weeks), smoke-exposure group 2 (24 weeks) and smoking cessation group, with 10 rats in each group. Alveolar airspace enlargement was observed by hematoxylin-eosin (HE) staining. IL-8 and TNF-α levels in bronchoalveolar lavage fluid (BALF) were tested by ELISA. The proportion of CD4(+)Foxp3(+) Treg in peripheral blood and lungs of rats was determined by flow cytometry. The mRNA expression of Foxp3 was measured by real-time PCR. Comparisons of the data between different groups were performed using one-way ANOVA, and SNK and Games-Howell test was used for comparison between 2 groups. RESULTS: The mean linear intercept (MLI) in smoke-exposure group 1 and group 2 [(64.9 ± 5.3) µm, (77.9 ± 11.5) µm] was higher than those in the control group 1 and group 2 [(39.0 ± 3.8) µm, (40.3 ± 2.7) µm], all P < 0.01. Compared with smoke-exposure group 2, the MLI in smoking cessation group (71.5 ± 5.8) µm showed a lower value (P < 0.01), but still higher than that in smoke-exposure group 1 (P < 0.01). The IL-8 and TNF-α levels in BALF of smoke-exposure group 1 and group 2 [(68 ± 17) ng/L, (85 ± 16) ng/L], [(14.1 ± 1.8) ng/L, (20.1 ± 8.7) ng/L] were higher than those in control group 1 and group 2 [(44 ± 8) ng/L, (43 ± 9) ng/L], [(6.3 ± 2.3) ng/L, (5.8 ± 1.6) ng/L], all P < 0.05. The IL-8 and TNF-α levels were not statistically different between in smoking cessation group (56 ± 6) ng/L, (14.7 ± 4.7) ng/L and smoke-exposure group 1. The percentage of Treg in the lungs of smoke-exposure group 1 and group 2 [(6.6 ± 0.8)%, (5.3 ± 0.9)%] was significantly decreased as compared to control group 1 and group 2 [(9.0 ± 1.0)%, (9.6 ± 0.9)%], all P < 0.01. The percentage of Treg in lungs was not statistically different between smoke-exposure group 1 and smoking cessation group (7.2 ± 0.6)%. In peripheral blood, there was no significant difference between groups in the percentage of Treg. In the lung, Foxp3 mRNA expression in smoke-exposure group 1 and group 2 [(17 ± 7), (9 ± 7)] was less than that in control group 1 and group 2 [(39 ± 6), (42 ± 7)], all P < 0.01. The Foxp3 mRNA expression was not statistically different between smoke-exposure group 1 and smoking cessation group (21 ± 9). No significant differences in peripheral blood Foxp3 mRNA expression was found between groups. CONCLUSIONS: Decreased Treg was present in lungs of cigarette smoke-induced model of emphysema despite 12 weeks' smoking cessation, suggesting that down-regulation of Treg may be involved in the amplified and persistent inflammation after smoking cessation.

Shikonin suppresses trophoblast cell growth via regulation of GLI1, and p62 mediated caspase 8 activation.

Unruptured ectopic pregnancy (UEP) is a common cause of morbidity and, occasionally, of mortality in women of reproductive age. Pharmacological intervention is a common therapeutic approach for early-stage UEP. Herein, we investigated the cytotoxic effect and novel mechanism of shikonin, a natural naphthoquinone pigment purified from Lithospermum erythrorhizon, in human trophoblast cells. These data demonstrated that shikonin suppressed proliferation and induced apoptosis in a time-dependent manner in HTR-8/SVneo cells. Shikonin blocked autophagic flux and promoted p62 interaction with caspase 8, resulting in caspase 8 activation. Moreover, shikonin suppressed GLI1 expression, and GLI1 overexpression attenuated shikonin-induced cell apoptosis. Although silencing GLI1 slightly promoted cell apoptosis, p62 overexpression enhanced GLI1 silencing-induced cell apoptosis by activating caspase 8. Furthermore, rapamycin increased shikonin-induced cell apoptosis in HTR-8/SVneo cells, whereas 3-MA attenuated the cytotoxic effect of shikonin. In conclusion, shikonin suppressed trophoblast cell growth by silencing GLI1 and increasing p62 co-mediated activation of caspase 8, which suggested a potential novel therapeutic target for UEP.

Thymosin beta4 is cytoprotective in human gingival fibroblasts.

Thymosin beta4 (Tbeta(4)) is a naturally occurring, ubiquitous, non-toxic protein with documented wound-healing, anti-inflammatory, anti-apoptotic, and tissue-repair properties in skin, the ocular surface, and the heart. The ability of Tbeta(4) to demonstrate similar protective properties in cells of the oral cavity was analyzed using an in vitro model of cultured human gingival fibroblasts. Thymosin beta 4 significantly suppressed the secretion of interleukin-8 (IL-8) following stimulation with tumor necrosis factoralpha (TNF-alpha), suggesting that it may suppress the inflammatory response initiated by pro-inflammatory cytokines. By contrast, Tbeta(4) was not effective in protecting fibroblasts from challenge with lipopolysaccharide purified from Porphyromonas gingivalis or Escherichia coli. Thymosin beta 4 was able to protect gingival fibroblasts against the known cytotoxic effects of chlorhexidine digluconate, a mouthrinse containing chlorhexidine digluconate, and carbamide peroxide. Additionally, Tbeta(4) was able to protect gingival fibroblasts from the apoptosis that is induced by stimulation with TNF-alpha or by exposure to chlorhexidine. Because of its multifunctional roles in protecting cells against damage, Tbeta(4) may have significant potential for use as an oral heathcare aid with combined antimicrobial, anti-inflammatory, anti-apoptotic, and cytoprotective properties.