An ensemble deep learning diagnostic system for determining Clinical Activity Scores in thyroid-associated ophthalmopathy: integrating multi-view multimodal images from anterior segment slit-lamp photographs and facial images.

Background: Thyroid-associated ophthalmopathy (TAO) is the most prevalent autoimmune orbital condition, significantly impacting patients' appearance and quality of life. Early and accurate identification of active TAO along with timely treatment can enhance prognosis and reduce the occurrence of severe cases. Although the Clinical Activity Score (CAS) serves as an effective assessment system for TAO, it is susceptible to assessor experience bias. This study aimed to develop an ensemble deep learning system that combines anterior segment slit-lamp photographs of patients with facial images to simulate expert assessment of TAO. Method: The study included 156 patients with TAO who underwent detailed diagnosis and treatment at Shanxi Eye Hospital Affiliated to Shanxi Medical University from May 2020 to September 2023. Anterior segment slit-lamp photographs and facial images were used as different modalities and analyzed from multiple perspectives. Two ophthalmologists with more than 10 years of clinical experience independently determined the reference CAS for each image. An ensemble deep learning model based on the residual network was constructed under supervised learning to predict five key inflammatory signs (redness of the eyelids and conjunctiva, and swelling of the eyelids, conjunctiva, and caruncle or plica) associated with TAO, and to integrate these objective signs with two subjective symptoms (spontaneous retrobulbar pain and pain on attempted upward or downward gaze) in order to assess TAO activity. Results: The proposed model achieved 0.906 accuracy, 0.833 specificity, 0.906 precision, 0.906 recall, and 0.906 F1-score in active TAO diagnosis, demonstrating advanced performance in predicting CAS and TAO activity signs compared to conventional single-view unimodal approaches. The integration of multiple views and modalities, encompassing both anterior segment slit-lamp photographs and facial images, significantly improved the prediction accuracy of the model for TAO activity and CAS. Conclusion: The ensemble multi-view multimodal deep learning system developed in this study can more accurately assess the clinical activity of TAO than traditional methods that solely rely on facial images. This innovative approach is intended to enhance the efficiency of TAO activity assessment, providing a novel means for its comprehensive, early, and precise evaluation.

Stigma predicting fertility quality of life among Chinese infertile women undergoing in vitro fertilization-embryo transfer.

OBJECTIVE: To investigate stigma and fertility quality of life (FertiQoL) and identify predictors of FertiQoL in Chinese infertile women undergoing in vitro fertilization-embryo transfer (IVF-ET). METHODS: A descriptive correlational design was adopted to investigate the association between stigma and FertiQoL in 588 infertile women undergoing IVF-ET. The personal information questionnaire, Infertility Stigma Scale (ISS) and FertiQoL tool were used to measure study variables. RESULTS: The mean scores of ISS and FertiQoL were 62.59 (SD = 21.58) and 63.64 (SD = 13.72), respectively. There were significant differences of ISS scores among participants with different educational level, residence, occupation, religious belief, financial condition, age group, duration of infertility and infertility treatment, while significant differences of the FertiQoL scores were found in participants with different insurance status, determinism of etiology, infertile type, duration of infertility treatment and cycles of IVF-ET. Pearson's correlation analysis showed stigma was negatively correlated with FertiQoL (r = -0.081 to -0.669, p < .05). The self-devaluation (ß = -0.290, p < .001), social withdrawal (ß = -0.237, p < .001), family stigma (ß = -0.217, p < .001) and insurance status (ß = 0.066, p=.035) were identified as the significant predictor of FertiQoL accounting for 43.5% of variance. CONCLUSIONS: The stigma was significantly associated with FertiQoL in infertile women undergoing IVF-ET with higher level of stigma predicting poorer FertiQoL. More psychological support should be provided to infertile women to reduce stigma and improve FertiQoL.

Isoelectric focusing on microfluidic paper-based chips.

Isoelectric focusing (IEF), a powerful technique for protein separation and enrichment, was successfully integrated into microfluidic paper-based analytical devices (µPADs) in this work. The µPADs for isoelectric focusing were fabricated by octadecyltrichlorosilane (OTS) silanization and subsequent region-selective plasma treatment. The system of IEF on µPADs could be easily assembled. And a series of conditions of the system were investigated, including the suitable concentration of ampholyte to create good pH gradient, the effect of polyvinylpyrrolidone (PVP) on electroosmotic flow (EOF) suppression, and focusing voltage applied on the paper channel. After optimization, simultaneous separation and enrichment of protein sample containing myoglobin and cytochrome C was successfully demonstrated. Besides, parallel IEF on multichannels were also achieved for the separation of multiple protein samples on one single chip, and their performance was compared with that of the conventional gel-IEF system. The developed IEF on µPADs exhibits appealing features such as low cost, simplicity, and disposability and are believed to have great application potentials.

Women with Recurrent Miscarriage Have Decreased Expression of 25-Hydroxyvitamin D3-1α-Hydroxylase by the Fetal-Maternal Interface.

BACKGROUND: Effects of vitamin D deficiency in pregnancy have been associated with some adverse pregnancy outcomes. The 25-hydroxyvitamin D3-1α-hydroxylase (CYP27B1) is integral to the vitamin D metabolic pathway. The enzyme catalyzes localized conversion of pro-hormone 25-hydroxyvitamin D3 to active 1,25-dihydroxyvitamin D3. Our aim was to investigate the expression of CYP27B1 at the fetal-maternal interface in the first trimester pregnancy and to determine whether CYP27B1 was associated with recurrent miscarriage (RM). METHODS: Expressions of CYP27B1 mRNA and protein in villi and decidua from 20 women undergoing primary miscarriage, 20 women with RM and 20 women with normal pregnancy were evaluated by western blot, and quantitative real-time PCR. The co-localization of CYP27B1 and certain cytokines including IL-10, IFN-γ, TNF-α, and IL-2 expression were examined using immunohistochemistry and confocal microscopy. RESULTS: Women with RM had a significantly lower expression of CYP27B1 mRNA and protein in villous and decidual tissues compared with the normal pregnant women (P = 0.000 in villus, P = 0.002 in decidua for mRNA; P = 0.036 in villus, P = 0.007 in decidua for protein.). Compared with the normal pregnancy, immunostaining for CYP27B1 was significantly decreased in villous trophoblasts and decidual glandular epithelial cells in RM women. No significant differences in the localization of CYP27B1, IL-10, IFN-γ, TNF-α, and IL-2 expression were identified between the normal pregnant and RM women. CONCLUSIONS: Women with RM have a lower level of CYP27B1 expression in chorionic villi and decidua compared with normal pregnant women, suggesting that reduced CYP27B1 expression may be associated with RM. The consistent localization of CYP27B1 and IL-10, IFN-γ, TNF-α, and IL-2 expression in villous and decidual tissues suggests the importance of the local production of 1,25(OH)2D3 at the fetal-maternal interface to regulate cytokine responses.

Decreased expression of the vitamin D receptor in women with recurrent pregnancy loss.

The multiple functions of vitamin D3 have stimulated interest in the role that this vitamin may play during pregnancy. The present study investigated the expression of the vitamin D receptor (VDR) in women during the first trimester of pregnancy in order to determine whether VDR is associated with recurrent pregnancy loss (RPL). Forty women at 7-10 weeks gestation with RPL and 40 women of similar gestational age with a healthy pregnancy were recruited. VDR mRNA and protein in chorionic villi and decidua were evaluated by immunohistochemistry, confocal laser scanning microscopy (CLSM), western blot, and quantitative real-time polymerase chain reaction. The serum levels of VDR were measured by an enzyme-linked immunosorbent assay. Women with RPL had a significantly weaker expression of VDR mRNA in villi and decidual tissues compared with the control women (both p < 0.0001). Western blot analysis showed an approximately 46% decrease in VDR expression in villi and a 52% decrease in decidua in the RPL vs. the controls. Serum VDR levels were also significantly lower in the RPL group than in the control group (p = 0.003). Compared with the controls, immunohistochemical and CLSM analysis revealed significantly lower VDR expression in villous cytotrophoblasts and stromal cells, as well as in decidual glandular epithelial and stromal cells (all p < 0.05). In conclusion, these observations show that women with RPL have lower levels of VDR expression in chorionic villi, decidua and serum compared with normal pregnant women, suggesting that decreased VDR expression in the first trimester pregnancy may be associated with RPL.

[ICSI with testicular or epididymal sperm for patients with obstructive azoospermia: A systematic review].

OBJECTIVE: To assess the effects of testicular sperm and epididymal sperm on the outcomes of ICSI for patients with obstructive azoospermia. METHODS: We searched PubMed, MEDLINE, EMBASE, Cochrane, CNKI, VIP, CBM, and Wanfang Database up to December 2015 for published literature relevant to ICSI with testicular or epididymal sperm for obstructive azoospermia patients. According to the inclusion and exclusion criteria, two reviewers independently conducted literature screening, data extraction and quality assessment of the included trials, followed by meta-analysis with the RevMan 5.3 software. RESULTS: A total of 14 studies were identified, involving 1 278 patients and 1 553 ICSI cycles. ICSI with epididymal sperm exhibited a significantly higher fertilization rate than that with testicular sperm (RR = 1.08, 95% CI 1.05－1.11, P<0.01). No statistically significant differences were observed between the epididymal and testicular sperm groups in the rates of cleavage (RR = 1.04, 95% CI 0.99－1.10, P = 0.13), good-quality embryo (RR = 1.01, 95% CI 0.93－1.09,P = 0.85), implantation (RR = 1.14, 95% CI 0.75－1.73, P = 0.55), clinical pregnancy (RR = 1.14, 95% CI 0.98－1.31, P = 0.08), and miscarriage (RR = 0.86, 95% CI 0.53－1.39,P = 0.54). CONCLUSIONS: ICSI with epididymal sperm yields a markedly higher fertilization rate than that with testicular sperm, but has no statistically significant differences from the latter in the rates of cleavage, good-quality embryo, implantation, clinical pregnancy, and miscarriage in the treatment of obstructive azoospermia.

Reduced CD200 and CD200R1 expression in human chorionic villi contributes to early spontaneous abortion.

OBJECTIVE: To examine the expression of CD200 and its receptor (CD200R) in human chorionic villi during the first trimester of normal pregnancy and early spontaneous abortion (ESA). DESIGN: Prospective study. METHODS: Expression of CD200 and CD200R in the chorionic villi was determined using streptavidin-peroxidase immunohistochemistry, confocal laser scanning microscopy and real-time polymerase chain reaction. POPULATION: Thirty-five women diagnosed with ESA and 30 women experiencing a healthy pregnancy in a medical university hospital in China were enrolled in this study between 2011 and 2013. MAIN OUTCOME MEASURES: CD200 and CD200R expression. RESULTS: The expression of CD200 in syncytiotrophoblast cells was significantly higher during normal pregnancy than in ESA (0.51 ± 0.05 vs. 0.35 ± 0.05). In contrast, expression of CD200 in cytotrophoblast cells and CD200R in stromal cells was significantly lower during normal pregnancy when compared with ESA (CD200: 0.16 ± 0.02 vs. 0.32 ± 0.03; CD200R: 0.19 ± 0.03 vs. 0.22 ± 0.02). In villi, the expression of both CD200 protein and CD200R transcripts were significantly higher in healthy first-trimester pregnancy than in ESA (CD200: 156.89 ± 105.65 vs. 37.51 ± 17.62). CONCLUSIONS: There is an increase in inhibitory properties of human chorionic villi during normal pregnancy. The mechanism underlying ESA might be associated with enhanced expression of CD200 and CD200R in the trophoblast, leading to an upregulation of the immune response during the first trimester of pregnancy.

SIRT1 inhibits TNF-α-induced apoptosis of vascular adventitial fibroblasts partly through the deacetylation of FoxO1.

Sirtuin 1 (SIRT1), a NAD(+)-dependent class III histone deacetylase, participates in regulating cellular apoptosis, senescence and metabolism by deacetylating histones and multiple transcription factors. In this study, we aimed to determine the effect of SIRT1 on the apoptosis of vascular adventitial fibroblasts (VAFs) and related signaling pathways. SIRT1 was found in the nucleus of VAFs and translocated into the cytoplasm in response to tumor necrosis factor-α (TNF-α). Moreover, SIRT1 protein expression was reduced in VAFs stimulated with TNF-α. In addition, TNF-α increased the apoptosis of VAFs. Activation of SIRT1 by resveratrol (RSV) or overexpression of SIRT1 attenuated TNF-α-induced VAF apoptosis by decreasing the percentage of apoptotic cells and cleaved caspase-3 protein expression and increasing the Bcl-2/Bax ratio. In contrast, inhibition of SIRT1 by sirtinol/nicotinamide or knockdown of SIRT1 enhanced apoptosis of VAFs. On the other hand, knockdown of FoxO1 reduced TNF-α-induced VAF apoptosis. SIRT1 interacted with FoxO1 in VAFs by the co-immunoprecipitation assay. Further study showed that RSV or SIRT1 overexpression decreased acetylated-FoxO1 (Ac-FoxO1) protein expression in VAFs stimulated with TNF-α. Knockdown of SIRT1 resulted in an increase in Ac-FoxO1 protein expression. Taken together, these findings indicate that SIRT1 inhibits the apoptosis of VAFs, whereas FoxO1 promotes VAF apoptosis. Furthermore, the inhibitory effect of SIRT1 on VAF apoptosis is partly mediated by the deacetylation of FoxO1.

SIRT1 regulates CD40 expression induced by TNF-α via NF-ĸB pathway in endothelial cells.

BACKGROUND: Compelling evidence suggests that SIRT1, NAD(+)-dependent class III protein deacetylase, plays an important role in the prevention and treatment of atherosclerosis by counteracting inflammation. Cluster of differentiation 40 (CD40), as a pro-inflammatory cytokine, has been shown to participate in the pathophysiology of atherosclerosis. The relationship between SIRT1 and CD40, however, remained elusive. The present study was thus designed to explore the potential effect of SIRT1 on CD40 expression induced by tumor necrosis factor-α (TNF-α) and to disclose the underlying mechanism in CRL-1730 endothelial cells. METHODS: mRNA and protein expressions were identified by quantitative real-time PCR and Western blot respectively. Subcellular localization of SIRT1 was detected by immunofluorescence analysis. SIRT1 small-interfering RNA (siRNA) was carried out for mechanism study. RESULTS: TNF-α reduced SIRT1 expression and induced CD40 expression in CRL-1730 endothelial cells in a time- and concentration- dependent manner. Pretreatment with resveratrol (a potent SIRT1 activator) inhibited TNF-α-induced CD40 expression, while pretreatment with nicotinamide (class b HDACs inhibitor nicotinamide) or sirtinol (a known SIRT1 inhibitor), especially SIRT1 siRNA significantly augmented TNF-α-induced CD40 expression. The frther sudy idicated that PDTC (NF-ĸB inhibitor) pretreatment attenuated TNF-α-induced CD40 expression, and SIRT1 siRNA significantly augmented TNF-α-induced acetylated-NF-ĸB p65 (Lys310) expression. CONCLUSION: The present study provides the direct evidence that SIRT1 can inhibit TNF-α- induced CD40 expression in CRL-1730 endothelial cells by deacetylating the RelA/p65 subunit of NF-ĸB at lysine 310, which provides new insights into understanding of the anti-inflammatory and anti-athroscerotic actions of SIRT1.

SIRT1 regulates TNF-α-induced expression of CD40 in 3T3-L1 adipocytes via NF-κB pathway.

Sirtuin1 (SIRT1), a NAD(+)-dependent deacetylase, not only regulates lipid and glucose homeostasis, but also involves the regulation of proinflammatory cytokine involved in inflammation-associated diseases. The activation of CD40 triggers inflammation that plays a crucial role in the development of many chronic inflammatory diseases including obesity. Growing evidence indicated that SIRT1 exerts anti-inflammatory properties by suppressing proinflammatory cytokines production. However, the effect of SIRT1 on the expression of CD40 in adipocytes has not yet been fully elucidated. The present study showed that SIRT1 expressed both in the nucleus and cytoplasm of 3T3-L1 adipocytes. TNF-α significantly reduced the expression of SIRT1 mRNA and protein and increased the expression of CD40 mRNA and protein in time- and concentration-dependent manners. Overexpression of SIRT1 or SIRT1 activation by resveratrol obviously attenuated the expression of CD40 induced by TNF-α in 3T3-L1 adipocytes, whereas knockdown of SIRT1 or SIRT1 inhibition by nicotinamide and sirtinol significantly enhanced TNF-α-induced expression of CD40. Furthermore, overexpression of SIRT1 or SIRT1 activation by resveratrol diminished TNF-α-induced acetylation of NF-κBp65, while knockdown of SIRT1 or SIRT1 inhibition by nicotinamide and sirtinol augmented TNF-α-induced acetylation of NF-κBp65 in 3T3-L1 adipocytes. NF-κB inhibitor PDTC reduced TNF-α-induced mRNA and protein expression of CD40 in 3T3-L1 adipocytes. The combination treatment of resveratrol and PDTC significantly reduced TNF-α-induced expression of CD40, and the inhibitory effects were higher than that of the single treatment. Taken together, SIRT1 exerts anti-inflammatory property by regulating TNF-α-induced expression of CD40 partially through the NF-κB pathway in 3T3-L1 adipocytes. More importantly, the regulation of SIRT1 on the expression of CD40 provides new insight to understand the anti-inflammatory effects of SIRT1.

Nuclear translocation of nuclear factor Kappa B in first trimester deciduas and chorionic villi in early spontaneous miscarriage women.

The nuclear factor kappa B is widely expressed in the distinct subpopulations of chorionic villi and deciduas of first-trimester pregnancies. We examined the cellular distribution and expression of nuclear factor kappa B in the human first-trimester chorionic villi and deciduas of women with early spontaneous miscarriage and viable pregnancy by confocal laser scanning microscope and immunohistochemistry. There is a greater nuclear translocation of nuclear factor kappa B is restricted to villous stromal cells, decidual stromal cells, glandular epithelial cells and vessel endothelial cells in early spontaneous miscarriage than in viable pregnancies. Collectively these observations suggest that over-activation of nuclear factor kappa B has a relationship with early spontaneous miscarriages.

Expression and localization of tumor necrosis factor receptor 1 protein in the chorionic villi in early normal and spontaneous abortion.

BACKGROUND: Increasing tumor necrosis factor alpha (TNFalpha) expression is found to be positively associated with spontaneous abortion. TNFalpha acts through the binding of two types of transmembrane receptors called TNFR1 and TNFR2. Here we compare membrane TNFR1 (mTNFR1) protein expression on chorionic villi in women with spontaneous abortion (SA) and viable pregnancy. METHODS: In a prospective study, 31 women with SA and 30 with a viable pregnancy were studied. Chorionic villous membrane TNFR1 was detected by confocal laser scanning microscopy and immunohistochemistry. RESULTS: The mTNFR1 protein is expressed in villous stromal, vessel endothelial, syncytiotrophoblast, and cytotrophoblast cells in early pregnancy. The intensity of mTNFR1 fluorescence (mean+/-S.D.) in villous stromal cells was higher in the abortion groups than in the control group (35.99+/-6.35 versus 32.64+/-5.01; p<0.05). In the abortion groups, villous cytotrophoblast cells were intensely positive for mTNFR1, whereas mTNFR1 staining of vessel endothelial and syncytiotrophoblast cells was significantly lower (p<0.001). CONCLUSION: Abundant mTNFR1 expression in the cytotrophoblast cells in women with SA may mediate TNFalpha to induce programmed cell death in the mTNFR1-expressing cytotrophoblasts to limit their growth. The over-expressing mTNFR1 in villous stromal cells, mediating TNFalpha effects, may cause pathological changes or tissue damage in chorionic villi locally in nonviable pregnancies.