Proteomic analysis reveals that cigarette smoke exposure diminishes ovarian reserve in mice by disrupting the CREB1-mediated ovarian granulosa cell proliferation-apoptosis balance.

Exposure to cigarette smoke (CS) adversely affects ovarian health and it is currently unknown how CS exposure causes ovarian injury. This study compared the differences in proteomics between CS exposure and healthy control groups using liquid chromatography-tandem mass spectrometry quantitative proteomics to further understand the molecular mechanism of ovarian cell injury in mice exposed to CS. Furthermore, western blotting and qPCR were carried out to validate the proteomic analysis outcomes. CREB1 was selected from the differentially expressed proteins, and then the down-regulation of CREB1 and phosphorylated CREB1(Ser133) expressions were confirmed in mice ovarian tissue and human ovarian granulosa cells (KGN cells) after CS exposure. In addition, the expressions of apoptosis-related proteins BCL-2 and BCL-XL were downregulated, and BAX expression was up-regulated. Moreover, the results of cellular immunofluorescence, flow cytometry, and transmission electron microscopy (TEM) showed that cigarette smoke extract (CSE) efficiently stimulated the production of reactive oxygen species, apoptosis, G1 phase arrest, mitochondrial membrane potential decreases, and ultrastructural changes in KGN cells. KG-501 (CREB inhibitor) aggravated CSE-induced mitochondrial dysfunction and apoptosis-proliferation imbalance in KGN cells mediated by down-regulated CREB1/BCL-2 axis. In addition, CREB1 over-expression partially restores mitochondrial dysfunction and apoptosis-proliferation imbalance of KGN cells induced by CSE. The results suggested that CSE diminished ovarian reserve in mice by disrupting the CREB1-mediated ovarian granulosa cell (GCs) proliferation-apoptosis balance and provided possible therapeutic targets for the clinical intervention of premature ovarian failure (POI) caused by CS exposure.

Untargeted metabolomics analysis reveals the metabolic disturbances and exacerbation of oxidative stress in recurrent spontaneous abortion.

BACKGROUND: Recurrent spontaneous abortion (RSA) is characterized by the occurrence of two or more consecutive spontaneous abortions, with a rising prevalence among pregnant women and significant implications for their physical and mental well-being. The multifaceted etiology of RSA has posed challenges in unraveling the molecular mechanisms underlying that underlie its pathogenesis. Oxidative stress and immune response have been identified as pivotal factors in the development of its condition. METHODS: Eleven serum samples from healthy pregnant women and 17 from RSA were subjected to liquid chromatography/mass spectrometry (LC-MS) analysis. Multivariate statistical analysis was employed to excavate system-level characterization of the serum metabolome. The measurement of seven oxidative stress products, namely superoxide dismutase (SOD), catalase (CAT), malonaldehyde (MDA), glutathione (GPx), glutathione peroxidase (GSH), oxidized glutathione (GSSG), heme oxygenase (HO-1), was carried out using ELISA. RESULTS: Through the monitoring of metabolic and lipid alternations during RSA events, we have identified 816 biomarkers that were implicated in various metabolic pathways, including glutathione metabolism, phosphonate and phosphinate metabolism, nucleotide metabolism, sphingolipid metabolism, lysine degradation and purine metabolism, etc. These pathways have been found to be closely associated with the progression of the disease. Our finding indicated that the levels of MDA and HO-1 were elevated in the RSA group compared to the control group, whereas SOD, CAT and GPx exhibited a contrary pattern. However, no slight difference was observed in GSH and GSSG levels between the RSA group and the control group. CONCLUSION: The manifestation of RSA elicited discernible temporal alternations in the serum metabolome and biochemical markers linked to the metabolic pathways of oxidative stress and immune response. Our investigation furnished a more comprehensive analytical framework encompassing metabolites and enzymes associated with oxidative stress. This inquiry furnished a more nuanced comprehension of the pathogenesis of RSA and established the ground work for prognostication and prophylaxis.

Contribution of Sucrose Metabolism in Phloem to Kiwifruit Bacterial Canker Resistance.

Kiwifruit bacterial canker, caused by Pseudomonas syringae pv. actinidiae (Psa), is a catastrophic disease affecting kiwifruit worldwide. As no effective cure has been developed, planting Psa-resistant cultivars is the best way to avoid bacterial canker in kiwifruit cultivation. However, the differences in the mechanism of resistance between cultivars is poorly understood. In the present study, five local kiwifruit cultivars were used for Psa resistance evaluation and classified into different resistance categories, tolerant (T), susceptible (S), and highly susceptible (HS), based on their various symptoms of lesions on the cane. Susceptible and highly susceptible varieties had a higher sucrose concentration, and a greater decrease in sucrose content was observed after Psa inoculation in phloem than in tolerant varieties. Three invertase activities and their corresponding gene expressions were detected in the phloem with lesions and showed the same trends as the variations in sucrose concentration. Meanwhile, after Psa inoculation, enzyme activities involved in antioxidant defense responses, such as PAL, POD, and CAT, were also altered in the phloem of the lesion position. With no differences among cultivars, PAL and POD activities in phloem first increased and then decreased after Psa inoculation. However, great differences in CAT activities were observed between T and S/HS categories. Our results demonstrate that sucrose content was negatively correlated with the disease resistance of different cultivars and that the increase in immune response enzymes is likely caused by increased sucrose metabolism in the phloem.

Single-nucleus RNA Sequencing reveals the mechanism of cigarette smoke exposure on diminished ovarian reserve in mice.

The systematic toxicological mechanism of cigarette smoke (CS) on ovarian reserve has not been extensively investigated. Female 8-week-old C57BL/6 mice at peak fertility were exposed to CS or indoor air only for 30 days (100 mice per group) and the effects of CS on ovarian reserve were assessed using Single-Nucleus RNA Sequencing (snRNA-seq). In addition, further biochemical experiments, including immunohistochemical staining, ELISA, immunofluorescence staining, transmission electron microscopy, cell counting kit-8 assay, flow cytometry analysis, senescence-associated ß-galactosidase staining, and western blotting, were accomplished to confirm the snRNA-seq results. We identified nine main cell types in adult ovaries and the cell-type-specific differentially expressed genes (DEGs) induced by CS exposure. Western blot results verified that down-regulation of antioxidant genes (Gpx1 and Wnt10b) and the steroid biosynthesis gene (Fdx1) occurred in both ovarian tissue and human granulosa cell-like tumor cell line (KGN cells) after CS exposure. Five percent cigarette smoke extract (CSE) effectively stimulated the production of reactive oxygen species (ROS), DNA damage, cellular senescence and markedly inhibited KGN cell proliferation by inducing G1-phase cell cycle arrest. Moreover, down-regulation of Gja1, Lama1 and the Ferroptosis indicator (Gpx4) in granulosa cells plays a significant role in ultrastructural changes in the ovary induced by CS exposure. These observations suggest that CS exposure impaired ovarian follicle reserve might be caused by REDOX imbalance in granulosa cells. The current study systematically determined the damage caused by CS in mouse ovaries and provides a theoretical basis for early clinical prediction, diagnosis and intervention of CS exposure-associated primary ovarian insufficiency (POI), and is of great significance in improving female reproductive health.

Fisetin-induced cell death in human ovarian cancer cell lines via zbp1-mediated necroptosis.

BACKGROUND: Among reproductive cancers, ovarian cancer leads to the highest female mortality rate. Fisetin, a natural flavonoid, exerts pharmacological effects, inhibiting cancer growth with various origins. Although multiple mechanisms are involved in regulating cell death, it is still unclear whether and how fisetin exhibits anticancer effects on ovarian cancer. The present study aimed to evaluate cell apoptotic and necroptotic processes occurring in ovarian carcinoma (OC) cell lines induced by fisetin. METHODS: Cell growth was evaluated by MTT assay in OC cell lines treated with or without fisetin. Annexin V/propidium iodide staining followed by flow cytometry was used to characterize fisetin-induced cell death. The apoptotic process was suppressed by z-VAD intervention, and cell necroptosis was assessed by introducing ZBP1-knockdown OC cell lines coupled with fisetin intervention. The expression of necroptosis-related mediators and the migration capability of the respective cells were evaluated by Western blotting and in vitro cell invasion assay. RESULT: Fisetin successfully reduced cell growth in both OC cell lines in a dose-dependent manner. Both apoptosis and necroptosis were induced by fisetin. Suppression of the cell apoptotic process failed to enhance the proliferation of fisetin-treated cells. The induced cell death and robust expression of the necroptotic markers RIP3 and MLKL were alleviated by knocking down the expression of the ZBP1 protein in both OC cell lines. CONCLUSION: The present study provided in vitro evidence supporting the involvement of both apoptosis and necroptosis in fisetin-induced OC cell death, while ZBP1 regulates the necroptotic process via the RIP3/MLKL pathway.

Mediation of mechanically adapted TiCu/TiCuN/CFR-PEEK implants in vascular regeneration to promote bone repair in vitro and in vivo.

Background/Objective: TiCu/TiCuN is a multilayer composite coating comprising TiN and Cu, which provides excellent wear resistance and antibacterial properties. However, its applicability as a functional coating has not been widely realised, and several aspects pertaining to its properties must still be explored. Methods: This study uses arc ion-plating technology to apply a TiCu/TiCuN coating on the surface of carbon fibre-reinforced (CFR) polyetheretherketone (PEEK) material.The safety and osteogenic activity of TiCu/TiCuN-coated CFR-PEEK materials were explored through cell experiments and animal experiments, and the molecules behind them were verified. Results: The new material exhibits improved mechanical compatibility (mechanical strength and elastic modulus) and superior light transmittance (elimination of metal artifacts and ray refraction during radiology and radiotherapy). The proposed implant delivers excellent biocompatibility for mesenchymal stem cells and human umbilical vein endothelial cells (HUVECs), and it exhibits excellent osteogenic activity both in vitro and in vivo. Additionally, it was determined that the applied TiCu/TiCuN coating aids in upregulating the expression of angiogenesis-related signals (i.e., cluster-of-differentiation 31, α-smooth muscle actin, vascular endothelial growth factor receptor, and hypoxia-inducible factor-1α) to promote neovascularisation, which is significant for characterising the mechanism of the coating in promoting bone regeneration. Conclusion: The current results reveal that the TiCu/TiCuN-coated CFR-PEEK implants may emerge as an advanced generation of orthopaedic implants. Translational potential statement: The results of this study indicate that TiCu/TiCuN coating-modified CFR-PEEK materials can promote bone repair through angiogenesis and have broad clinical translation prospects.

[Phenotypic analysis and variant identification of a fetus with Joubert syndrome 17].

OBJECTIVE: To analyze the phenotype and genetic variant of a fetus with dysplasia of cerebellar vermis. METHODS: Gestational status and family history of the gravida was taken in combination with the imaging results of the fetus. Following elected abortion, fetal tissue and peripheral blood samples of the couple were collected for the extraction of genome DNA. Whole exome sequencing was carried out to screen potential variant associated with the phenotype of the proband. Specific PCR primers were designed to verify the results by Sanger sequencing. RESULTS: Prenatal ultrasound revealed that the fetal vermis cerebellum was poorly developed, which was similar to the previous pregnancy. Whole exome sequencing revealed that the fetus has carried compound heterozygous variants of the CPLANE1 gene, namely c.7978C>T and c.7169delT, which were respectively inherited from the husband and wife. CONCLUSION: The c.7978C>T and c.7169delT compound heterozygous variants of the CPLANE1 gene probably underlay the dysplasia of cerebellar vermis in the fetus, which has provided a basis for genetic counseling and prenatal diagnosis.

Improved diagnosis of thyroid cancer aided with deep learning applied to sonographic text reports: a retrospective, multi-cohort, diagnostic study.

OBJECTIVE: Large volume radiological text data have been accumulated since the incorporation of electronic health record (EHR) systems in clinical practice. We aimed to determine whether deep natural language processing algorithms could aid radiologists in improving thyroid cancer diagnosis. METHODS: Sonographic EHR data were obtained from the EHR database. Pathological reports were used as the gold standard for diagnosing thyroid cancer. We developed thyroid cancer diagnosis based on natural language processing (THCaDxNLP) to interpret unstructured sonographic text reports for thyroid cancer diagnosis. We used the area under the receiver operating characteristic curve (AUROC) as the primary metric to measure the performance of the THCaDxNLP. We compared the performance of thyroid ultrasound radiologists aided with THCaDxNLP vs. those without THCaDxNLP using 5 independent test sets. RESULTS: We obtained a total number of 788,129 sonographic radiological reports. The number of thyroid sonographic data points was 132,277, 18,400 of which were thyroid cancer patients. Among the 5 test sets, the numbers of patients per set were 439, 186, 82, 343, and 171. THCaDxNLP achieved high performance in identifying thyroid cancer patients (the AUROC ranged from 0.857-0.932). Thyroid ultrasound radiologists aided with THCaDxNLP achieved significantly higher performances than those without THCaDxNLP in terms of accuracy (93.8% vs. 87.2%; one-sided t-test, adjusted P = 0.003), precision (92.5% vs. 86.0%; P = 0.018), and F1 metric (94.2% vs. 86.4%; P = 0.007). CONCLUSIONS: THCaDxNLP achieved a high AUROC for the identification of thyroid cancer, and improved the accuracy, sensitivity, and precision of thyroid ultrasound radiologists. This warrants further investigation of THCaDxNLP in prospective clinical trials.

Circadian transcription factor Dbp promotes rat calvarial osteoprogenitors osteogenic differentiation through Kiss1/GnRH/E2 signaling pathway loop.

To determine the mechanism by which D-site-binding protein (Dbp) regulates rat calvarial osteoprogenitors (OPCs) osteogenic differentiation. α-Smooth muscle actin (α-SMA) + rat calvarial OPCs were extracted and purified using immunomagnetic beads. Cells were transduced with Dbp-lentivirus and divided into Dbp knockdown, Dbp overexpression and vehicle groups. After osteogenic induction for 21 days, Alizarin red staining and alkaline phosphatase (ALP) activity were examined. Expression levels of Runx2, Ocn, Osterix, Bmp4, Kiss1, and GnRH were determined using a quantitative real-time polymerase chain reaction. The observed changes in Kisspeptin, GnRH, ERα, and Runx2 were further validated via Western blot analysis. Furthermore, E2 and GnRH secretion levels were detected via an enzyme-linked immunosorbent assay (ELISA). Chromatin immunoprecipitation (ChIP) and luciferase assay were used to assess the effects of Dbp on the Kiss1 gene promoter. The coexpression of Dbp and Kisspeptin or GnRH was also evaluated via immunofluorescence. Following osteogenic induction, Dbp overexpression significantly increased calcium nodule formation and ALP activity, as well as Runx2, Ocn, Osterix, Bmp4, Kiss1, and GnRH messenger RNA expression, while Dbp knockdown presented the opposite results. Western blot analysis and ELISA results showed that Dbp significantly promotes Runx2, E2/ERα, Kisspeptin, and GnRH expression. These findings were confirmed by the ChIP assay, which indicated that the estrogen receptor promotes Kisspeptin expression after binding to the Kiss1 gene promoter, which is regulated by Dbp. Immunofluorescence assay showed that Dbp coexpression with Kisspeptin or GnRH varied depending on Dbp expression levels. Collectively, the circadian transcription factor Dbp promotes α-SMA + rat calvarial OPCs osteoblastic differentiation through Kiss1/GnRH/E2 signaling pathway loop.

Effect of ciRS-7 expression on clear cell renal cell carcinoma progression.

BACKGROUND: Circular RNA ciRS-7 has been reported to be involved in the progression of various cancers. However, ciRS-7 expression and its role in clear cell renal cell carcinoma (ccRCC) progression remains unclear. This study aimed to investigate the effect of ciRS-7 expression on ccRCC and the related signaling pathway. METHODS: ciRS-7 expression was analyzed using quantitative reverse transcription polymerase chain reaction in 87 pairs of ccRCC and matched adjacent normal tissues. The role of ciRS-7 in ccRCC cell proliferation and invasion was determined using the cell counting kit-8 and invasion assays, respectively. Potential mechanisms underlying the role of ciRS-7 in promoting ccRCC progression were explored by Western blotting. The relationship between the expression of ciRS-7 and features of ccRCC was analyzed by the Chi-square test and progression-free survival was determined using a Kaplan-Meier plot. RESULTS: ciRS-7 was overexpressed in ccRCC tissues compared with that in matched adjacent normal tissues. In addition, ciRS-7 up-regulation was closely associated with tumor diameter (P = 0.050), clinical stage (P = 0.009), and distant metastasis (P = 0.007). ciRS-7 knockdown in 786O and 769P cells markedly inhibited their proliferative and invasive abilities. In addition, ciRS-7 inhibition reduced phosphorylated epidermal growth factor receptor (p-EGFR) and phosphorylated serine/threonine kinase (p-Akt) levels. CONCLUSIONS: ciRS-7 up-regulation could promote ccRCC cell proliferation and invasion, which may be related with the EGFR/Akt signaling pathway. ciRS-7 might be a potential ccRCC therapeutic target.

Spontaneous rupture of an unscarred uterus with an intact amniotic sac extrusion and fetal leg entrapment at 28 gestational weeks: a case report.

The spontaneous rupture of an unscarred uterus at 28 gestational weeks is an extremely rare event, particularly when associated with an intact amniotic sac extrusion and fetal leg entrapment, which has not been previously reported. A 27-year-old primigravid woman was referred to our department, due to perpetual abdominal pain, at 28 weeks and 5 days of gestation. The patient, G3p0, had previously undergone two induced abortions. At the time of admission, abdominal ultrasonography suggested a defect in the left uterine horn. An emergency laparotomy was subsequently performed and revealed an intact amniotic sac extrusion and fetal leg entrapment. Considering the risk of placental abruption, and the possibility of a secondary rupture if the gestation was not terminated, an emergency Cesarean section was recommended. Uterine rupture may be suspected whenever a patient complains of durative abdominal pain at 28 weeks and 5 days of gestation, even in the absence of an intra-abdominal hemorrhage or vaginal bleeding.

Effects of microplastic biofilms on nutrient cycling in simulated freshwater systems.

Microplastic surfaces could be colonized by microorganisms and form biofilms in aquatic ecosystem, which can participate in the nitrogen (N) and phosphorus (P) cycles. In this work, polypropylene squares were deployed in a pond for 30 days for microplastic biofilms colonization and then were transported to indoor microcosms at an environmental relevant level to study their effects on N and P cycling. Results showed that microplastic biofilms could accelerate ammonia and nitrite oxidation as well as denitrification. Presence of microplastic biofilms accumulated P temporarily and increased alkaline phosphatase activities (APA) in the system. Later in the experiment, disintegration of matured biofilms released N and P into the water. Mass balance calculation suggested possible N input caused by biological nitrogen fixation. Our results demonstrated that microplastics associated biofilms have the ability to alter the N and P cycling processes in aquatic system. However, additional works are required to further quantify the extent of such impact.

Surgical site infection following operative treatment of open fracture: Incidence and prognostic risk factors.

Considering the high incidence of postoperative complications of open fracture, management of this injury is an intractable challenge for orthopaedist, and surgical site infection (SSI) is the devastate one. Screening for high-risk patients and target them with appropriate interventions is important in clinical practice. The aim of this study was to identify modifiable factors that were associated with SSI following operative treatment of open fractures. This retrospective, multicentre study was conducted at three hospitals. A total of 2692 patients with complete data were recruited between June 2015 and July 2018. Demographic characteristics, operation relative variables, additional comorbidities, and biochemical indexes were extracted and analysed. Receiver operating characteristic analysis was performed to detect the optimum cut-off value for some variables. Univariate and multivariate logistic analysis models were performed, respectively, to identify the independent risk factors of SSI. The overall incidence of SSI was 18.6%, with 17.0% and 1.6% for superficial and deep infection, respectively. Results of univariate and multivariate analyses showed the following: fracture type, surgical duration > 122 minutes, anaesthesia time > 130 minutes, intraoperative body temperature < 36.4°C, blood glucose (GLU) > 100 mg/dL, blood platelet (PLT) < 288 × 109 , and white blood cells (WBC) > 9.4 × 109 were independent risk factors of postoperative wound infection following operative treatment of open fractures. Six modifiable factors such as surgical duration > 122 minutes, anaesthesia time > 130 minutes, intraoperative body temperature < 36.4°C, GLU > 100 mg/dL, PLT < 288 × 109, and WBC > 9.4 × 109 play an important role in the prevention of SSI, and these factors should be optimized perioperatively.

Homeobox A10 promotes the proliferation and invasion of bladder cancer cells via regulation of matrix metalloproteinase-3.

Homeobox A10 (HOXA10) belongs to the family of HOX genes, which are closely connected with embryonic development and serve important roles in various tumors. However, the role of HOXA10 in bladder cancer (BC) remains unclear. In the present study, the role of HOXA10 in BC and the underlying mechanisms by which it promotes the disease progression were investigated. Immunohistochemical analysis demonstrated that the expression of the HOXA10 protein was significantly higher in BC tissues as compared with that in adjacent normal tissues. Subsequent statistical analysis revealed that upregulation of HOXA10 was significantly associated with the pathological grade and clinical stage of BC patients. In the BC cell lines T24 and 5637, silencing of HOXA10 by small interfering RNA transfection suppressed the proliferation, migration and invasion of BC cells, and led to decreased matrix metalloproteinase-3 expression. Taken together, overexpression of HOXA10 may be associated with poor prognosis in BC, and may serve as a novel antitumor therapy target for the treatment of this disease.

Serum Metabolomics Study of Nonsmoking Female Patients with Non-Small Cell Lung Cancer Using Gas Chromatography-Mass Spectrometry.

The incidence of nonsmoking female patients with non-small cell lung cancer (NSCLC) has increased in recent decades; however, the pathogenesis of patients is unclear, and early diagnosis biomarkers are in urgent need. In this study, 136 nonsmoking female subjects (65 patients with NSCLC, 6 patients with benign lung tumors, and 65 healthy controls) were enrolled, and their metabolic profiling was investigated by using pseudotargeted gas chromatography-mass spectrometry. A total of 56 annotated metabolites were found and verified to be significantly different in nonsmoking females with NSCLC compared with the control. The metabolic profiling was featured by disturbed energy metabolism, amino acid metabolism, oxidative stress, lipid metabolism, and so on. Cysteine, serine, and 1-monooleoylglycerol were defined as the biomarker panel for the diagnosis of NSCLC patients. 98.5 and 91.4% of subjects were correctly distinguished in the discovery and validation sets, respectively. The biomarker panel was also useful for the diagnosis of in situ malignancy patients, with an accuracy of 97.7 and 97.8% in the discovery and validation sets, respectively. The study provides a biomarker panel for the auxiliary diagnosis of nonsmoking females with NSCLC.

Diagnosis of thyroid cancer using deep convolutional neural network models applied to sonographic images: a retrospective, multicohort, diagnostic study.

BACKGROUND: The incidence of thyroid cancer is rising steadily because of overdiagnosis and overtreatment conferred by widespread use of sensitive imaging techniques for screening. This overall incidence growth is especially driven by increased diagnosis of indolent and well-differentiated papillary subtype and early-stage thyroid cancer, whereas the incidence of advanced-stage thyroid cancer has increased marginally. Thyroid ultrasound is frequently used to diagnose thyroid cancer. The aim of this study was to use deep convolutional neural network (DCNN) models to improve the diagnostic accuracy of thyroid cancer by analysing sonographic imaging data from clinical ultrasounds. METHODS: We did a retrospective, multicohort, diagnostic study using ultrasound images sets from three hospitals in China. We developed and trained the DCNN model on the training set, 131 731 ultrasound images from 17 627 patients with thyroid cancer and 180 668 images from 25 325 controls from the thyroid imaging database at Tianjin Cancer Hospital. Clinical diagnosis of the training set was made by 16 radiologists from Tianjin Cancer Hospital. Images from anatomical sites that were judged as not having cancer were excluded from the training set and only individuals with suspected thyroid cancer underwent pathological examination to confirm diagnosis. The model's diagnostic performance was validated in an internal validation set from Tianjin Cancer Hospital (8606 images from 1118 patients) and two external datasets in China (the Integrated Traditional Chinese and Western Medicine Hospital, Jilin, 741 images from 154 patients; and the Weihai Municipal Hospital, Shandong, 11 039 images from 1420 patients). All individuals with suspected thyroid cancer after clinical examination in the validation sets had pathological examination. We also compared the specificity and sensitivity of the DCNN model with the performance of six skilled thyroid ultrasound radiologists on the three validation sets. FINDINGS: Between Jan 1, 2012, and March 28, 2018, ultrasound images for the four study cohorts were obtained. The model achieved high performance in identifying thyroid cancer patients in the validation sets tested, with area under the curve values of 0·947 (95% CI 0·935-0·959) for the Tianjin internal validation set, 0·912 (95% CI 0·865-0·958) for the Jilin external validation set, and 0·908 (95% CI 0·891-0·925) for the Weihai external validation set. The DCNN model also showed improved performance in identifying thyroid cancer patients versus skilled radiologists. For the Tianjin internal validation set, sensitivity was 93·4% (95% CI 89·6-96·1) versus 96·9% (93·9-98·6; p=0·003) and specificity was 86·1% (81·1-90·2) versus 59·4% (53·0-65·6; p<0·0001). For the Jilin external validation set, sensitivity was 84·3% (95% CI 73·6-91·9) versus 92·9% (84·1-97·6; p=0·048) and specificity was 86·9% (95% CI 77·8-93·3) versus 57·1% (45·9-67·9; p<0·0001). For the Weihai external validation set, sensitivity was 84·7% (95% CI 77·0-90·7) versus 89·0% (81·9-94·0; p=0·25) and specificity was 87·8% (95% CI 81·6-92·5) versus 68·6% (60·7-75·8; p<0·0001). INTERPRETATION: The DCNN model showed similar sensitivity and improved specificity in identifying patients with thyroid cancer compared with a group of skilled radiologists. The improved technical performance of the DCNN model warrants further investigation as part of randomised clinical trials. FUNDING: The Program for Changjiang Scholars and Innovative Research Team in University in China, and National Natural Science Foundation of China.

Associations of IL-1, 6, and 10 Gene Polymorphisms with Susceptibility to Recurrent Aphthous Stomatitis: Insights from a Meta-Analysis.

AIM: To determine if there are significant associations between polymorphisms of the IL-1, IL-6, and IL-10 genes and susceptibility to recurrent aphthous stomatitis (RAS). METHODS: The PubMed, Embase, and Web of Science databases were searched for all eligible studies using both medical subheadings and free terms through December 2016. A total of 226 citations were retrieved. Odds ratios were used to quantitatively evaluate the associations of IL-1, IL-6, and IL-10 gene polymorphisms with RAS risk. A meta-analysis was performed, and heterogeneity, sensitivity, and subgroup analyses were carried out to clarify and validate the pooled results. RESULTS: A total of 11 studies were identified that met the inclusion criteria and were included in the meta-analysis. This current systematic review indicated that the IL-1b+3954 C/T polymorphism was significantly associated with an elevated risk of RAS onset for all inheritance models, except for the dominant model. For the IL-10-592 C/A polymorphism, protective associations with RAS were found using both the additive and recessive models, while it increased the risk of RAS in the codominant model. In Asian populations, the IL-10-1082 G/A polymorphism was associated with a protective effect for RAS using the allelic, additive, and recessive models. The IL-6-174 G/C polymorphism was not statistically associated with RAS risk. CONCLUSION: The IL-1b+3954 C/T polymorphism significantly increases RAS risk. In addition, the IL-10-1082 G/A polymorphism provided protective effects for RAS in the Asian population.

Induction of reprogramming of human amniotic epithelial cells into iPS cells by overexpression of Yap, Oct4, and Sox2 through the activation of the Hippo-Yap pathway.

The present study has reported a novel method for producing induced pluripotent stem (iPS) cells. Primary human amniotic epithelial cells (HuAECs) were isolated from the amniotic membranes of pregnant women who received Cesarean sections. These cells were infected with retroviruses carrying octamer-binding transcription factor 4 (Oct4), (sex determining region Y)-box 2 (Sox2) and Yes-associated protein (Yap) (OSY). Following in vitro culture for ~14 days, epithelial-like HuAECs exhibited several iPS clone-like cell colonies (OSY-iPS). These cell clones presented positive alkaline phosphatase features and expressed high levels of embryonic stem cell-like markers (Nanog homeobox, Sox2, Oct4, reduced expression protein 1, and SSES3/4). Additionally, epigenetic analysis results indicated that the methylation of CpG islands on endogenous Oct4 and Sox2 promoters was reduced in OSY-iPS cells. Furthermore, the majority of the histone H3 at lysine 9 sites that interacted with the Oct4 and Sox2 promoters were acetylated, suggesting that the transcription activities of the above two transcription factors significantly increased. In vivo and in vitro induced differentiation experiments demonstrated that OSY-iPS could develop into embryoid bodies in vitro, and express numerous cellular markers in the three germ layers. Furthermore, OSY-iPS could form teratomas in immunodeficient mice. The pathological detection results suggest that these teratomas contain numerous types of cells from the three germ layers. However, the results from the quantitative polymerase chain reaction and western blot analyses suggest that the Hippo-Yap signaling pathway was significantly activated in OSY-iPS cells. In conclusion, a novel method for iPS induction was established in the present study. HuAECs were successfully induced to reprogram iPS cells through the introduction of OSY to activate the Hippo-Yap signaling pathway.

Overexpression of SOX18 promotes prostate cancer progression via the regulation of TCF1, c-Myc, cyclin D1 and MMP-7.

Sex determining region Y (SRY)-box 18 (SOX18) gene encodes transcription factors that have been recently confirmed to be overexpressed in various human types of cancer and maintain the malignant behavior of cancer cells. However, the role and its potential function in prostate cancer (PCa) has not been demonstrated and the mechanisms of SOX18 involved in tumor progression remain largely unclear. In the present study, the expression of SOX18 was analyzed in 98 PCa and 81 adjacent non-tumor tissues using immunohistochemistry. The data showed that SOX18 was overexpressed in 72 of 98 (73.5%) PCa tissues compared with that in 28 of 81 (34.6%) non-tumor tissues. In addition, the expression of SOX18 was related with the clinical features of patients with PCa. To explore the potential role of SOX18 in PCa cells, Cell Counting Kit-8 (CCK-8), migration, invasion and xenograft assays were performed. Our data showed that knockdown of SOX18 decreased the proliferation, migration and invasion of PCa cells in vitro, in addition to the tumor growth in vivo. Markedly, SOX18 knockdown caused the decreased expression of TCF1, c-Myc, cyclin D1 and MMP-7. In conclusion, SOX18 was overexpressed in PCa and may regulate the malignant capacity of cells via the upregulation of TCF1, c-Myc, cyclin D1 and MMP-7.

Successful Experience of Laparoscopic Pancreaticoduodenectomy and Digestive Tract Reconstruction With Minimized Complications Rate by 14 Case Reports.

Laparoscopic pancreatic surgery is one of the most sophisticated and advanced applications of laparoscopy in the current surgical practice. The adoption of laparoscopic pancreaticoduodenectomy (LPD) has been relatively slow due to the technical challenges. The aim of this study is to review and characterize our successful LPD experiences in patients with distal bile duct carcinoma, periampullary adenocarcinoma, pancreas head cancer, and duodenal cancer and evaluate the clinical outcomes of LPD for its potential in oncologic surgery applications.We retrospectively analyzed the clinical data from 14 patients who underwent LPD from August 2013 to February 2015 in our institute.We presented our LPD experience with no cases converted to open surgery in all 14 cases, which included 10 cases of laparoscopic digestive tract reconstruction and 4 cases of open digestive tract reconstructions. There were no deaths during the perioperative period and no case of gastric emptying disorder or postoperative bleeding. The other clinical indexes were comparable to or better than open surgery.Based on our experience, LPD could be potentially safe and feasible for the treatment of early pancreas head cancer, distal bile duct carcinoma, periampullary adenocarcinoma, and duodenal cancer. The master of LPD procedure requires technical expertise but it can be accomplished with a short learning curve.