Distribution and Risk Assessment of Organophosphate Esters in Agricultural Soils and Plants in the Coastal Areas of South China.

Organophosphate esters (OPEs) are frequently used as flame retardants and plasticizers in various commercial products. While initially considered as substitutes for brominated flame retardants, they have faced restrictions in some countries due to their toxic effects on organisms. We collected 37 soil and crop samples in 20 cities along the coast of South China, and OPEs were detected in all of them. Meanwhile, we studied the contamination and potential human health risks of OPEs. In soil samples, the combined concentrations of eight OPEs varied between 74.7 and 410 ng/g, averaging at 255 ng/g. Meanwhile, in plant samples, the collective concentrations of eight OPEs ranged from 202 to 751 ng/g, with an average concentration of 381 ng/g. TDCIPP, TCPP, TCEP, and ToCP were the main OPE compounds in both plant and soil samples. Within the study area, the contaminants showed different spatial distributions. Notably, higher OPEs were found in coastal agricultural soils in Guangdong Province and crops in the Guangxi Zhuang Autonomous Region. The results of an ecological risk assessment show that the farmland soil along the southern coast of China is at high or medium ecological risk. The average non-carcinogenic risk and the carcinogenic risk of OPEs in soil through ingestion and dermal exposure routes are within acceptable levels. Meanwhile, this study found that the dietary intake of OPEs through food is relatively low, but twice as high as other studies, requiring serious attention. The research findings suggest that the human risk assessment indicates potential adverse effects on human health due to OPEs in the soil-plant system along the coast of South China. This study provides a crucial foundation for managing safety risks in agricultural operations involving OPEs.

Atypical glandular cells in Pap tests: cytology-histology correlation and risk assessment with human papillopmavirus (HPV) testing.

INTRODUCTION: Atypical glandular cells (AGC) represent less than 1% of Pap test cases and include a variety of lesions in both the cervix and endometrium. The study aimed to investigate the cytology-histology correlation in AGC patients and to evaluate the clinical utility of hrHPV testing in this diagnostic context. MATERIALS AND METHODS: We identified 491 atypical glandular cells (AGC) cases in our quality analysis (QA) database of 336,064 Pap tests interpreted between March 1, 2013 and July 12, 2016. Of these, 251 cases had follow-up biopsies with hrHPV tests in 148 cases. RESULTS: The most common histologic diagnosis associated with AGC was normal/benign or low-grade lesions, comprising 55% of cervical biopsies and 24% of endometrial biopsies. High-grade lesions were identified in 21% of follow-up biopsies. In patients with AGC cytology, a positive hrHPV test significantly increased the likelihood of cervical HSIL or above lesions on biopsy by 26.4 times (OR = 26.4, 95% CI: 5.8-119.4, P < 0.0001). A positive genotyping result for HPV 16 dramatically increased the likelihood of cervical HSIL or above lesions on biopsy (OR = 84, 95% CI: 12.0-590.5, P < 0.0001). The HPV test had a negative predictive value of 97% (CI: 85%-100%). CONCLUSIONS: Our study confirms that AGC is a significant diagnosis with an overall risk for high-grade cervical or endometrial lesions as high as 21%. hrHPV testing with genotyping is an effective tool for identifying high-risk individuals within the AGC population, with excellent positive and negative predictive values. This approach is valuable for clinical risk stratification and differential diagnosis in patients with AGC cytology.

Clinical significance of microscopic polyangiitis with interstitial lung disease and bronchiectasis: probability of preexisting comorbidities.

BACKGROUND: The association between pulmonary involvement and microscopic polyangiitis (MPA) has been increasingly recognized in recent years. Whether interstitial lung disease (ILD) and bronchiectasis (BE) are disease manifestations of MPA, preexisting comorbidities or important complications remains unclear. The purpose of this study was to determine the clinical characteristics and prognosis of MPA with pulmonary involvement to further guide clinical management. METHODS: The data for 97 patients with a definitive diagnosis of MPA were retrospectively reviewed. The MPA diagnosis was based on the 2012 revised Chapel Hill Consensus Conference (CHCC) criteria. The baseline clinical information and laboratory parameters were collected and analysed at each patient's initial diagnosis. RESULTS: Forty-seven out of the 97 (48.5%) patients who were diagnosed with MPA presented with pulmonary involvement, including 37 patients with ILD, 12 patients with BE and two patients with diffuse alveolar haemorrhage (DAH). ILD and BE antedated MPA in 56.76% and 75.00% of the patients, respectively. Compared with that in the MPA-BE group, the serum LDH level (222.86 ± 68.19 vs. 171.58 ± 31.43, p = .016) in the MPA-ILD group was significantly higher. In the multivariate Cox analysis, elevated serum creatinine (HR 4.08, confidence interval (CI) 1.38-12.05, p = .011) was an independent risk factor for shorter survival in MPA patients with pulmonary involvement, and treatment with glucocorticoid pulse cyclophosphamide therapy (HR 0.095, 95% CI 0.019-0.47, p = .004) was independently associated with prolonged survival. Among the patients in the MPA-ILD group, acute exacerbations of ILD (HR 4.55 CI 1.16-17.86, p = .029) and elevated serum creatinine (HR 4.95, CI 1.39-17.54, p = .014) were independently associated with a poor prognosis, and treatment with glucocorticoids (HR 0.057, 95% CI 0.012-0.28, p < .001) was independently associated with significant prolongation of survival. CONCLUSIONS: Patients with MPA have a high prevalence of pulmonary involvement, and ILD is the most common subtype of MPA. ILD and BE can be considered preexisting comorbidities of MPA. Elevated serum creatinine was associated with shorter survival. However, remission induction regimens with glucocorticoids and/or immunosuppressants may improve this outcome.

In-depth study of anticancer drug diffusion through a cross-linked -pH-responsive polymeric vesicle membrane.

Post-encapsulation and release of the anticancer drug doxorubicin hydrochloride (DOX·HCl) through cell-like transmission functions of polymeric vesicles were studied using cross-linked pH-responsive polymeric vesicles. The vesicles were fabricated for the first time via the redox-initiated reversible addition-fragmentation chain transfer dispersion polymerization in ethanol-water mixture, using 2-(diisopropylamino)ethyl methacrylate and glycidyl methacrylate, and the vesicle membrane was modified post-cross-linking by using ethylenediamine. A phase diagram was constructed for reproducible fabrication of the polymeric vesicles, and well-shaped vesicles were formed when the target degree of polymerization of the hydrophobic polymer chains was equal to or higher than 50 with solid content in the range of 10-30 wt%. The cross-linked vesicle membrane served as a gate enabling "open" and "closed" states in response to pH stimulation. Up to 50% drug loading efficiency and 39% drug loading content could be achieved, and in vitro release of the DOX-loaded vesicles in aqueous buffer solutions showed a much faster DOX release rate at pH 5.0 than at pH 6.5. The polymeric vesicles were of very low cytotoxicity to A549 cells up to the concentration of 2 mg/mL, and the IC50 of DOX-loaded vesicles were higher than that of the free DOX. The intracellular DOX release study indicated higher cellular uptake capability for DOX-loaded vesicles than that of free DOX.

Levels, Distribution and Health Risk Assessment of Organochlorine Pesticides in Agricultural Soils from the Pearl River Delta of China.

To reveal the pollution status of agricultural soils along with rapid urbanization and economic growth, a large regional survey of organochlorine pesticides (OCPs) in agricultural soils was conducted in the Pearl River Delta (PRD) of China. The results showed that the total residues of 23 OCPs were in the range of ND-946 ng/g dry weight. OCP residues showed distinct spatial distribution characteristics within the PRD. OCPs were mainly found in areas with high agricultural production and industrial activities. Higher OCP concentrations were observed in the top layer of soil, while the concentration decreases to marginal levels when the soil depth is greater than 50 cm. OCPs are mainly derived from historical use. Hexachlorocyclohexanes (HCHs) in the top soil of the study area are mainly from the use of lindane. Soil pH was negatively and significantly correlated with total OCP concentration. The human health risk assessment showed no health risk for children, while for adults, there is a non-carcinogenic risk, which needs to be noticed. Agricultural activities and industrial production have made the region a pollution hotspot and should arouse more stringent regulation to protect the environment and food safety.

Aptamer Targets Triple-Negative Breast Cancer through Specific Binding to Surface CD49c.

Although targeted cancer therapy can induce higher therapeutic efficacy and cause fewer side effects in patients, the lack of targetable biomarkers on triple-negative breast cancer (TNBC) cells limits the development of targeted therapies by antibody technology. Therefore, we investigated an alternative approach to target TNBC by using the PDGC21T aptamer, which selectively binds to poorly differentiated carcinoma cells and tumor tissues, although the cellular target is still unknown. We found that synthetic aptamer probes specifically bound cultured TNBC cells in vitro and selectively targeted TNBC xenografts in vivo. Subsequently, to identify the target molecule on TNBC cells, we performed aptamer-mediated immunoprecipitation in lysed cell membranes followed by liquid chromatography tandem mass spectrometry (LC-MS/MS). Sequencing analysis revealed a highly conserved peptide sequence consistent with the cell surface protein CD49c (integrin α3). For target validation, we stained cultured TNBC and non-TNBC cells with an aptamer probe or a CD49c antibody and found similar cell staining patterns. Finally, competition cell-binding assays using both aptamer and anti-CD49c antibody revealed that CD49c is the biomarker targeted by the PDGC21T aptamer on TNBC cells. Our findings provide a molecular foundation for the development of targeted TNBC therapy using the PDGC21T aptamer as a targeting ligand.

miR-582-5p inhibits migration and chemo-resistant capabilities of colorectal cancer cells by targeting TNKS2.

BACKGROUND: Metastasis and chemo-resistance are still important factors that limit the overall efficacy of colorectal cancer treatment. Understanding the detailed molecular mechanism and identifying potential biomarkers are of great value in prognosis prediction and risk stratification. OBJECTIVE: We investigated the role of miR-582-5p in colorectal cancer pathogenesis, progression and chemo-resistance. Furthermore, we explored the underlying molecular mechanism of miR-582-5p in modulation of malignant behaviors of colorectal cancer cells. METHODS: Clinical samples and colorectal cancer cell lines were applied to explore miR-582-5p expression level and its significance on tumor cell metastasis and chemo-resistance. Transwell study and cellular survivability study were performed to explore the influences of miR-582-5p expression modulation on tumor cell chemo-resistance and invasion/migration. Dual-luciferase reporter gene assay was conducted to explore the influences of miR-582-5p on its target gene TNKS2. RESULTS: Colorectal cancer patients with lymph node or distal organ metastatic diseases exhibited significantly lower level of miR-582-5p. In vitro studies have indicated that miR-582-5p inhibition significantly increased migration and chemo-resistant capabilities of tumor cells. And dual-luciferase reporter gene assay demonstrated that miR-582-5p exhibited its influences on the biological behavior of tumor cells by targeting TNKS2. CONCLUSIONS: Our study demonstrated for the first time that miR-582-5p played an important role for colorectal tumor cell metastasis and chemo-resistance. Our research also indicated that miR-582-5p and its target gene TNKS2 could be novel biomarkers for metastatic disease prediction, overall prognosis evaluation, as well as potential therapeutic target for colorectal cancer patients.

[Expression of TGF-ß in bladder neck contracture after transurethral enucleation and resection of the prostate].

OBJECTIVE: To explore the role of transforming growth factor-ß (TGF-ß) in bladder neck contracture (BNC) after transurethral enucleation and resection of the prostate (TUERP). METHODS: This study included 300 BPH patients undergoing TUERP, aged 51－89 (69.19 ± 8.43) years, with the prostate volume of 14.4－355.8 (63.18 ± 47.63) ml and preoperative IPSS of 15－35 (26.07 ± 5.9), QOL score of 3－6 (4.43 ± 0.67), PSA content of 0.17－23.16 (2.94 ± 3.77) ug/L, urinary leukocyte increase in 50 cases, post-void residual urine volume (PVR) of 0－440 (83.53 ± 86.85) ml, and maximum urinary flow rate (Qmax) of 2.3－14.5 (7.77 ± 3.47) ml/s. During TUERP, we collected the tissues from the bladder neck at 5 and 7 o'clock as well as the BPH tissue and the tissue from the residual prostate for HE staining, immunohistochemistry (the SP method) and examination of the infiltration degree of inflammatory cells and expressions of TGF-ß1 and TGF-ß3. During the 6－24 months follow-up, 6 of the patients were confirmed with BNC based on the clinical symptoms and the results of uroflowmetry and cystoscopy, and underwent transurethral bladder neck incision and detection of the expressions of TGF-ß1 and TGF-ß3 in the bladder neck tissue with BNC. RESULTS: The bladder neck tissue without BNC was mainly composed of smooth muscle and fibrous tissues with local infiltration of inflammatory cells, and the residual prostate tissue primarily comprised fibrous and muscle tissues, mixed with a little prostatic epithelial tissue. The bladder neck tissue with BNC, compared with that harvested during the initial TUERP, exhibited significantly increased expression of TGF-ß1 (ï¼»68.20 ± 10.88ï¼½% vs ï¼»36.14 ± 7.62ï¼½%, P < 0.05), decreased expression of TGF-ß3 (ï¼»8.55 ± 4.73ï¼½% vs ï¼»20.77 ± 8.69ï¼½%, P < 0.05), and enhanced infiltration of inflammatory cells (P < 0.05). The bladder neck tissue without BNC, in comparison with the BPH tissue, showed dramatically up-regulated expressions of TGF-ß1 (ï¼»27.05 ± 8.21ï¼½% vs ï¼»1.61 ± 0.69ï¼½%, P < 0.001) and TGF-ß3 (ï¼»14.09 ± 4.19ï¼½% vs ï¼»0.32 ± 0.11ï¼½%, P < 0.001) and increased infiltration of inflammatory cells (P < 0.05). CONCLUSIONS: After TUERP, the expression of TGF-ß1 is increased, that of TGF-ß3 decreased and the infiltration of inflammatory cells enhanced in the bladder neck tissue with BNC, which suggests that BNC may be related to the expression of TGF-ß and that BNC after TUERP could be prevented by regulating the expression of TGF-ß.

MnO2-coated porous Pt@CeO2 core-shell nanostructures for photoacoustic imaging-guided tri-modal cancer therapy.

We describe the synthesis of MnO2-coated porous Pt@CeO2 core-shell nanostructures (Pt@CeO2@MnO2) as a new theranostic nano-platform. The porous Pt cores endow the core-shell nanostructures with high photothermal conversion efficiency (80%) in the near-infrared region, allowing for photothermal therapy (PTT) and photoacoustic imaging (PA) of tumors. The combination of the Pt core and porous CeO2 interlayer enhances the separation of photo-generated electrons and holes, which is beneficial for the generation of singlet oxygen. With the porous structures of the cores and interlayers, the Pt@CeO2@MnO2 nanostructures are further loaded with an anti-cancer drug (doxorubicin, DOX). The degradation of the MnO2 shell in the tumor microenvironment (TME) can generate O2 for enhanced photodynamic therapy (PDT) and simultaneously trigger DOX release. PA imaging shows good accumulation and retention of DOX-loaded Pt@CeO2@MnO2 in tumors, which guides precise laser irradiation to initiate combined PTT and PDT. The synergistic PTT/PDT/chemotherapy demonstrated by DOX-loaded Pt@CeO2@MnO2 yields remarkable therapeutic outcomes in vitro and in vivo. Taken together, our DOX-loaded Pt@CeO2@MnO2 provides a new avenue for designing high-performance nano-platforms for imaging and therapeutics.

Quiescin sulfhydryl oxidase 1 promotes sorafenib-induced ferroptosis in hepatocellular carcinoma by driving EGFR endosomal trafficking and inhibiting NRF2 activation.

Sorafenib is a first-line molecular-target drug for advanced hepatocellular carcinoma (HCC), but its clinical effects are still limited. In this study we identify Quiescin sulfhydryl oxidase 1 (QSOX1) acting as a cellular pro-oxidant, specifically in the context of sorafenib treatment of HCC. QSOX1 disrupts redox homoeostasis and sensitizes HCC cells to oxidative stress by inhibiting activation of the master antioxidant transcription factor NRF2. A negative correlation between QSOX1 and NRF2 expression was validated in tumor tissues from 151 HCC patients. Mechanistically, QSOX1 restrains EGF-induced EGFR activation by promoting ubiquitination-mediated degradation of EGFR and accelerating its intracellular endosomal trafficking, leading to suppression of NRF2 activity. Additionally, QSOX1 potentiates sorafenib-induced ferroptosis by suppressing NRF2 in vitro and in vivo. In conclusion, the data presented identify QSOX1 as a novel candidate target for sorafenib-based combination therapeutic strategies in HCC or other EGFR-dependent tumor types.

Predict multicategory causes of death in lung cancer patients using clinicopathologic factors.

BACKGROUND: Random forests (RF) is a widely used machine-learning algorithm, and outperforms many other machine learning algorithms in prediction-accuracy. But it is rarely used for predicting causes of death (COD) in cancer patients. On the other hand, multicategory COD are difficult to classify in lung cancer patients, largely because they have multiple labels (versus binary labels). METHODS: We tuned RF algorithms to classify 5-category COD among the lung cancer patients in the surveillance, epidemiology and end results-18, whose lung cancers were diagnosed in 2004, for the completeness in their follow-up. The patients were randomly divided into training and validation sets (1:1 and 4:1 sample-splits). We compared the prediction accuracy of the tuned RF and multinomial logistic regression (MLR) models. RESULTS: We included 42,257 qualified lung cancers in the database. The COD were lung cancer (72.41%), other causes or alive (14.43%), non-lung cancer (6.85%), cardiovascular disease (5.35%), and infection (0.96%). The tuned RF model with 300 iterations and 10 variables outperformed the MLR model (accuracy = 69.8% vs 64.6%, 1:1 sample-split), while 4:1 sample-split produced lower prediction-accuracy than 1:1 sample-split. The top-10 important factors in the RF model were sex, chemotherapy status, age (65+ vs < 65 years), radiotherapy status, nodal status, T category, histology type and laterality, all of which except T category and laterality were also important in MLR model. CONCLUSION: We tuned RF models to predict 5-category CODs in lung cancer patients, and show RF outperforms MLR in prediction accuracy. We also identified the factors associated with these COD.

QSOX1 promotes mitochondrial apoptosis of hepatocellular carcinoma cells during anchorage-independent growth by inhibiting lipid synthesis.

Anoikis is a programmed death of cell induced upon detachment from the extracellular matrix (ECM). Resistance to anoikis is a critical contributor to cancer invasion and metastasis. High frequency of metastatic recurrence is a huge challenge for current therapy of hepatocellular carcinoma (HCC). Our previous study had identified sulfhydryl oxidase 1 (QSOX1) as a suppressor of HCC metastasis. In the present study, we used the anchorage-independent growth condition to mimic the detachment of HCC cells from ECM. We found that QSOX1 was induced in HCC cells under the anchorage-independent growth condition and that could be blocked by endoplasmic reticulum stress (ERS) inhibitor. Overexpression and knockdown of QSOX1 gene were performed on HCC cells. QSOX1 inhibited de novo synthesis of fatty acids (FAs) and cholesterol (ChE) and reduced their content in the detached HCC cells, and thus mediated mitochondrial apoptosis of HCC cells. In conclusion, QSOX1 is induced under detached culture condition via ERS. QSOX1 promotes mitochondrial apoptosis by suppressing the lipid synthesis of HCC cells in detached condition. QSOX1 appears to accelerate anoikis of HCC cells. These findings offer a new insight into how to overcome anoikis resistance of HCC cells and provide a potential target for prevention of HCC metastasis.

Chloride Distribution and Steel Corrosion in a Concrete Bridge after Long-Term Exposure to Natural Marine Environment.

Chloride-induced steel corrosion is the most concerning issue for the durability of concrete structures. Concrete and steel samples were obtained from a 30-year-old reinforced concrete bridge. The chloride content was measured by a potentiometric titration method and the microstructure of concrete was obtained by scanning electron microscopy and mercury intrusion porosimetry. The rust phases of the steel were detected by X-ray diffraction and Raman analysis. It was found that the convection depth for chloride transport in cracked concrete was significantly larger than that in uncracked concrete. The concrete in a pier column facing upstream had greater porosity due to the water impact and calcium leaching. The coefficients of variability of chloride diffusivity of concrete for the bridge deck and the pier column were significantly different. Rust phases including lepidocrocite, goethite, akaganeite, magnetite, and maghemite were detected using Raman spectroscopy and X-ray diffraction. The major phases of steel rust in the atmospheric zone were lepidocrocite and goethite, while they were lepidocrocite and maghemite in the tidal zone. The results of this study would provide information concerning the chloride-induced steel corrosion under a marine environment in order to predict long-term behaviors of a reinforced concrete structure.

Disruption of SIRT7 Increases the Efficacy of Checkpoint Inhibitor via MEF2D Regulation of Programmed Cell Death 1 Ligand 1 in Hepatocellular Carcinoma Cells.

BACKGROUND & AIMS: Immune checkpoint inhibitors have some efficacy in the treatment of hepatocellular carcinoma (HCC). Programmed cell death 1 ligand 1 (PD-L1), expressed on some cancer cells, binds to the receptor programmed cell death 1 (PDCD1, also called PD1) on T cells to prevent their proliferation and reduce the antigen-tumor immune response. Immune cells that infiltrate some types of HCCs secrete interferon gamma (IFNG). Some HCC cells express myocyte enhancer factor 2D (MEF2D), which has been associated with shorter survival times of patients. We studied whether HCC cell expression of MEF2D regulates expression of PD-L1 in response to IFNG. METHODS: We analyzed immune cells from 20 fresh HCC tissues by flow cytometry. We analyzed 225 fixed HCC tissues (from 2 cohorts) from patients in China by immunohistochemistry and obtained survival data. We created mice with liver-specific knockout of MEF2D (MEF2DLPC-KO mice). We knocked out or knocked down MEF2D, E1A binding protein p300 (p300), or sirtuin 7 (SIRT7) in SMMC-7721, Huh7, H22, and Hepa1-6 HCC cell lines, some incubated with IFNG. We analyzed liver tissues from mice and cell lines by RNA sequencing, immunoblot, dual luciferase reporter, and chromatin precipitation assays. MEF2D protein acetylation and proteins that interact with MEF2D were identified by coimmunoprecipitation and pull-down assays. H22 cells, with MEF2D knockout or without (controls), were transplanted into BALB/c mice, and some mice were given antibodies to deplete T cells. Mice bearing orthotopic tumors grown from HCC cells, with or without knockout of SIRT7, were given injections of an antibody against PD1. Growth of tumors was measured, and tumors were analyzed by immunohistochemistry and flow cytometry. RESULTS: In human HCC specimens, we found an inverse correlation between level of MEF2D and numbers of CD4+ and CD8+ T cells; level of MEF2D correlated with percentages of PD1-positive or TIM3-positive CD8+ T cells. Knockout of MEF2D from H22 cells reduced their growth as allograft tumors in immune-competent mice but not in immune-deficient mice or mice with depletion of CD8+ T cells. When MEF2D-knockout cells were injected into immune-competent mice, they formed smaller tumors that had increased infiltration and activation of T cells compared with control HCC cells. In human and mouse HCC cells, MEF2D knockdown or knockout reduced expression of PD-L1. MEF2D bound the promoter region of the CD274 gene (encodes PD-L1) and activated its transcription. Overexpression of p300 in HCC cells, or knockout of SIRT7, promoted acetylation of MEF2D and increased its binding, along with acetylated histones, to the promoter region of CD274. Exposure of HCC cells to IFNG induced expression of p300 and its binding MEF2D, which reduced the interaction between MEF2D and SIRT7. MEF2D-induced expression of PD-L1 upon IFNG exposure was independent of interferon-regulatory factors 1 or 9. In HCC cells not exposed to IFNG, SIRT7 formed a complex with MEF2D that attenuated expression of PD-L1. Knockout of SIRT7 reduced proliferation of HCC cells and growth of tumors in immune-deficient mice. Compared with allograft tumors grown from control HCC cells, in immune-competent mice, tumors grown from SIRT7-knockout HCC cells expressed higher levels of PD-L1 and had reduced infiltration and activation of T cells. In immune-competent mice given antibodies to PD1, allograft tumors grew more slowly from SIRT7-knockout HCC cells than from control HCC cells. CONCLUSIONS: Expression of MEF2D by HCC cells increases their expression of PD-L1, which prevents CD8+ T-cell-mediated antitumor immunity. When HCC cells are exposed to IFNG, p300 acetylates MEF2D, causing it to bind the CD274 gene promoter and up-regulate PD-L1 expression. In addition to promoting HCC cell proliferation, SIRT7 reduced acetylation of MEF2D and expression of PD-L1 in HCC cells not exposed to IFNG. Strategies to manipulate this pathway might increase the efficacy of immune therapies for HCC.

Age-specific 3-year cumulative risk of cervical cancer and high-grade dysplasia on biopsy in 9434 women who underwent HPV cytology cotesting.

BACKGROUND: High-risk human papillomavirus (HPV)-Papanicolaou (Pap) cotesting is recommended for cervical cancer screening in women aged ≥30 years. The current study analyzed the effectiveness of cotesting on risk management in different age groups. METHODS: A retrospective review of a 5-year cytology database identified 9434 women with HPV-Pap cotesting and follow-up cervical biopsy. The 3-year cumulative risk of developing high-grade cervical lesions (≥high-grade squamous intraepithelial lesion [HSIL]) was analyzed using age stratification. RESULTS: The 3-year cumulative risk of developing ≥HSIL was found to be significantly different in women with baseline cotesting HPV-positive and Pap-positive results (HPV+/Pap+; defined as ≥atypical squamous cells of undetermined significance), HPV+ and Pap-negative results, and HPV-negative and Pap+ results at 19.2%, 7.9%, and 3.1%, respectively (P < .001). The risk of ≥HSIL peaked at ages 30 to 39 years and significantly decreased at ages 50 to 59 years (16.6% vs 6.7%; P < .001). Women aged <30 years shared a high risk similar to that of women aged 30 to 39 years (17.3% vs 16.6%; P = .52), and risk stratification by cotesting was found to be equally effective in the younger age group (HPV+ and Pap+: 19.6%; HPV+ and Pap-negative: 7.2%; and HPV-negative and Pap+: 4.4% [P < .001]). CONCLUSIONS: High-risk HPV-Pap cotesting appears to be extremely sensitive for the prediction of the risk of developing ≥HSIL and is an effective tool for risk stratification. In the current study, the 3-year cumulative risk of developing ≥HSIL varied significantly with age, with the highest risk noted among women aged <40 years and the lowest risk observed in women aged 50 to 59 years. Pap testing significantly impacted risk stratification in the HPV+ positive group, especially in women aged <60 years. Women aged <30 years were found to have a risk profile and cotesting efficacy similar to those of women aged 30 to 39 years. Modification of the current recommendation to offer cotesting to women aged ≥30 years might be considered to include those patients aged <30 years.

Rapid Detection of Mycoplasma-Infected Cells by an ssDNA Aptamer Probe.

Mycoplasmas are unique cell wall-free bacteria. Because they lack a cell wall and have resistance to ß-lactam antibiotics, mycoplasma is the major pathogen that infects cultured cells in research laboratories. For rapid detection of mycoplasma-infected cells, we developed an ssDNA aptamer sequence composed of 40 nucleotides. Flow cytometry analysis showed that the synthetic aptamer probe selectively targeted mycoplasma-infected culture cells with high specificity identical to commercially available PCR-based assays. Additionally, fluorescent microscopy studies revealed that the aptamer probe rapidly stained mycoplasma-infected cells with higher sensitivity compared to Hoechst dye-mediated cellular DNA content stains. Moreover, confocal microscopy studies of trypsin-treated cells validated that the aptamer probes selectively targeted mycoplasma components on the surface of infected cells. Finally, preclinical studies of peripheral blood cells demonstrated that the aptamer probe was able to detect in vitro mycoplasma infection of primary lymphocytes. Taken together, these findings indicate that the aptamer probe will not only allow rapid detection of mycoplasma-infected culture cells for research purposes but also provide a simple method to monitor mycoplasma infection in primary cell products for clinical use.

High GCLC level in tumor tissues is associated with poor prognosis of hepatocellular carcinoma after curative resection.

Glutamate-cysteine ligase catalytic subunit (GCLC) has been reported to overexpress in a variety types of cancer and be related with tumor progression and drug resistance. However, little has been known about GCLC's prognostic significance and biological roles in hepatocellular carcinoma (HCC). In the present study, we evaluated GCLC expression level using immunohistochemical staining (IHC) in tissue microarray (TMA) containing paired tumor and peritumoral liver tissues from 168 patients with HCC who received curative resection. GCLC levels in tumor tissues were significantly higher than in peritumoral liver tissues, and tumor GCLC level was associated with overall survival (OS) and disease-free survival (DFS). Five-year OS and DFS rates were 41.15% and 25.88% for the group with high tumor GCLC level, compared with 68.09% and 47.51% for the group with low tumor GCLC level (P<0.001 and P=0.001, respectively). Moreover, quantitative reverse transcription PCR (qRT-PCR) analysis demonstrated that GCLC was transcriptionally activated in HCC tissues when comparing with peritumoral tissues. Tumor GCLC level, which correlated to tumor differentiation, microvascular invasion and BCLC stage, was independent prognostic factors for both OS (P=0.006) and DFS (P=0.003). Importantly, tumor GCLC level was still significantly associated with OS and DFS in patients with early HCC. GCLC-based nomogram models were further established and exhibit significantly higher predictive accuracy as compared with routine clinical staging systems. In conclusion, tumor GCLC is a potential prognostic biomarker for HCC patients after receiving curative resection.

Tcf7l1 Acts as a Suppressor for the Self-Renewal of Liver Cancer Stem Cells and Is Regulated by IGF/MEK/ERK Signaling Independent of ß-Catenin.

Tcf7l1, which is a key effector molecule of the Wnt/ß-catenin signaling pathway, is highly expressed in various cancers, and it promotes tumor growth. In this study, we demonstrated that unlike its tumor-promoting effects in several other types of cancers, Tcf7l1 expression is downregulated in hepatocarcinoma compared with their adjacent nontumor counterparts. Underexpression of Tcf7l1 is correlated with poorer survival. In liver cancer stem cell (CSC) populations, Tcf7l1 expression is downregulated. Ectopic expression of Tcf7l1 attenuates the self-renewal abilities of liver CSCs. Mechanistically, Tcf7l1 regulates the self-renewal abilities of liver CSCs through transcriptional repression of the Nanog gene, and the effect is independent of ß-catenin. Moreover, we found that Tcf7l1 expression is controlled by extracellular insulin-like growth factor (IGF) signaling, and we demonstrated for the first time that IGF signaling stimulates Tcf7l1 phosphorylation and degradation through the mitogen-activated protein kinase (MEK)/extracellular signal-regulated kinase (ERK) pathway. Overall, our results provide some new insights into how extracellular signals modulate the self-renewal of liver CSCs and highlight the inhibitory roles of Tcf7l1 in cancer. Stem Cells 2019;37:1389-1400.

Primary adrenal gland epithelioid sarcoma: A case report and literature review.

Epithelioid sarcoma is a malignant mesenchymal neoplasm with morphologic and immunophenotypic epithelioid differentiation, which rarely arises in solid organs. We report a case of primary epithelioid sarcoma in the adrenal gland of a 31-year-old female. The patient initially presented with nausea and rectal bleeding, and subsequent imaging studies revealed a 4.4 cm left adrenal gland mass and left retroperitoneal lymphadenopathy. Clinical and radiological studies did not reveal tumor elsewhere in the patient. Histologic features were those of epithelioid sarcoma, proximal type with cohesive clusters of epithelioid tumor cells harboring frequent mitoses, and areas of necrosis. Immunohistochemical stains showed strong, diffuse expression of epithelial markers (pancytokeratin), and CD34 and Fli-1. Partial and focal positive staining of CK7 was also noted. Nuclear expression of SMARCB1 (INI-1) protein was lost. ERG was negative in this case. We believe that this is the second-case report of a primary adrenal gland epithelioid sarcoma. Fli-1 positivity was seen in our case, and ERG was negative as shown in some recent publications regarding epithelioid sarcomas.