Cell of origin alters myeloid-mediated immunosuppression in lung adenocarcinoma.

Solid carcinomas are often highly heterogenous cancers, arising from multiple epithelial cells of origin. Yet, how the cell of origin influences the response of the tumor microenvironment is poorly understood. Lung adenocarcinoma (LUAD) arises in the distal alveolar epithelium which is populated primarily by alveolar epithelial type I (AT1) and type II (AT2) cells. It has been previously reported that Gramd2 + AT1 cells can give rise to a histologically-defined LUAD that is distinct in pathology and transcriptomic identity from that arising from Sftpc + AT2 cells1,2. To determine how cells of origin influence the tumor immune microenvironment (TIME) landscape, we comprehensively characterized transcriptomic, molecular, and cellular states within the TIME of Gramd2 + AT1 and Sftpc + AT2-derived LUAD using KRASG12D oncogenic driver mouse models. Myeloid cells within the Gramd2 + AT1-derived LUAD TIME were increased, specifically, immunoreactive monocytes and tumor associated macrophages (TAMs). In contrast, the Sftpc + AT2 LUAD TIME was enriched for Arginase-1+ myeloid derived suppressor cells (MDSC) and TAMs expressing profiles suggestive of immunosuppressive function. Validation of immune infiltration was performed using flow cytometry, and intercellular interaction analysis between the cells of origin and major myeloid cell populations indicated that cell-type specific markers SFTPD in AT2 cells and CAV1 in AT1 cells mediated unique interactions with myeloid cells of the differential immunosuppressive states within each cell of origin mouse model. Taken together, Gramd2 + AT1-derived LUAD presents with an anti-tumor, immunoreactive TIME, while the TIME of Sftpc + AT2-derived LUAD has hallmarks of immunosuppression. This study suggests that LUAD cell of origin influences the composition and suppression status of the TIME landscape and may hold critical implications for patient response to immunotherapy.

A cuproptosis-related signature predicts prognosis and indicates cross-talk with immunocyte in ovarian cancer.

PURPOSE: Cuproptosis, programmed cell death by intracellular copper-mediated lipoylated protein aggregation, is involved in various tumorigenesis and drug resistance abilities by mediating the tumor microenvironment. Previous studies have demonstrated that serum copper levels are higher in OC patients than in normal subjects. However, the exact relationship between cuproptosis and ovarian cancer progression remains to be further elucidated. METHODS: The Cancer Genome Atlas (TCGA) and gene expression omnibus (GEO) datasets were utilized to establish a cuproptosis-related prognostic signature in ovarian cancer. Subsequently, the bulk RNA-seq analysis and single-cell RNA-seq analysis were used to identify the relationship between signature with immune cell infiltration, chemotherapy, and cuproptosis-related scoring (CuRS) system. Finally, the potential biological functional roles of target genes in cuproptosis were validated in vitro. RESULTS: By using LASSO-Cox regression analysis to establish the cuproptosis-related prognostic model, our works demonstrated the accuracy and efficiency of our model in the TCGA (583 OC patients) and GEO (260 OC patients) OC cohorts, and the high-scoring groups showed worse survival outcomes. Notably, there were substantial differences between the high and low-risk groups in extensive respects, such as the activating transcription factors, cell pseudotime features, cell intercommunication patterns, immunocytes infiltration, chemotherapy response, and potential drug resistance. KIF26B was selected to construct a prognostic model from the identified 33 prognosis-related genes, and high expression of KIF26B predicted poorer prognosis in ovarian cancer. Ultimately, further in vitro experiments demonstrated that KIF26B participated in the proliferation and cisplatin resistance of OC cells. Knockdown of KIF26B increased the sensitivity of OC cells to elesclomol, a cuproptosis agonists. CONCLUSION: This study constructed a new cuproptosis-related gene signature that has a good prognostic capacity in assessing the outcome of OC patients. This study enhances our understanding of cuproptosis associated with ovarian cancer aggressiveness, cross-talk with immunocytes, and serves as a novel chemotherapy strategy.

Suvemcitug plus chemotherapy for platinum-resistant epithelial ovarian, fallopian tube and primary peritoneal cancer: A phase 1b dose-escalation trial.

OBJECTIVE: The use of bevacizumab has been hampered by safety concerns despite demonstrable progression-free survival (PFS) benefit in subjects with platinum-resistant ovarian cancer, highlighting the need for novel effective and safe antiangiogenic agents. This study aimed to characterize the tolerability, safety, and antitumor activities of escalating doses of anti-VEGF antibody suvemcitug plus chemotherapy in platinum-resistant ovarian cancer patients. METHODS: This open-label, dose-escalation trial enrolled adult patients (≥18 years) with platinum-resistant histologically or cytologically-confirmed epithelial ovarian, fallopian tube and primary peritoneal cancer. Eligible patients received paclitaxel or topotecan plus escalating doses of suvemcitug 0.5, 1, 1.5, or 2 mg/kg once every two weeks. The primary endpoints were safety and tolerability, and antitumor activities of suvemcitug. RESULTS: Twenty-nine subjects received paclitaxel (n = 11) or topotecan (n = 18). No dose-limiting toxicities occurred. The most common adverse events of special interest were proteinuria (41.4%), hypertension (20.7%) and epistaxis (10.3%). No gastrointestinal perforations occurred. Nine subjects (31.0%, 95% CI 15.3-50.8) demonstrated investigators-confirmed objective response, including complete response in 1 and partial response in 8. The median PFS was 5.4 months (95% CI 2.2-7.4). CONCLUSIONS: Suvemcitug demonstrated an acceptable safety profile and promising antitumor activities in platinum-resistant ovarian cancer patients, supporting its further clinical development.

M2 macrophage-derived exosomal circTMCO3 acts through miR-515-5p and ITGA8 to enhance malignancy in ovarian cancer.

Tumor-associated macrophages of the M2 phenotype promote cancer initiation and progression. Importantly, M2 macrophage-derived exosomes play key roles in the malignancy of cancer cells. Here, we report that circTMCO3 is upregulated in ovarian cancer patients, and its high expression indicates poor survival. M2-derived exosomes promote proliferation, migration, and invasion in ovarian cancer, but these effects are abolished by knockdown of circTMCO3. Furthermore, circTMCO3 functions as a competing endogenous RNA for miR-515-5p to reduce its abundance, thus upregulating ITGA8 in ovarian cancer. miR-515-5p inhibits ovarian cancer malignancy via directly downregulating ITGA8. The decreased oncogenic activity of circTMCO3-silencing exosomes is reversed by miR-515-5p knockdown or ITGA8 overexpression. Exosomal circTMCO3 promotes ovarian cancer progression in nude mice. Thus, M2 macrophage-derived exosomes promote malignancy by delivering circTMCO3 and targeting the miR-515-5p/ITGA8 axis in ovarian cancer. Our findings not only provide mechanistic insights into ovarian cancer progression, but also suggest potential therapeutic targets.

Inhibition of USP7 upregulates USP22 and activates its downstream cancer-related signaling pathways in human cancer cells.

Deubiquitinases (DUBs) play important roles in various human cancers and targeting DUBs is considered as a novel anticancer therapeutic strategy. Overexpression of ubiquitin specific protease 7 and 22 (USP7 and USP22) are associated with malignancy, therapy resistance, and poor prognosis in many cancers. Although both DUBs are involved in the regulation of similar genes and signaling pathways, such as histone H2B monoubiquitination (H2Bub1), c-Myc, FOXP3, and p53, the interdependence of USP22 and USP7 expression has never been described. In the study, we found that targeting USP7 via either siRNA-mediated knockdown or pharmaceutical inhibitors dramatically upregulates USP22 in cancer cells. Mechanistically, the elevated USP22 occurs through a transcriptional pathway, possibly due to desuppression of the transcriptional activity of SP1 via promoting its degradation upon USP7 inhibition. Importantly, increased USP22 expression leads to significant activation of downstream signal pathways including H2Bub1 and c-Myc, which may potentially enhance cancer malignancy and counteract the anticancer efficacy of USP7 inhibition. Importantly, targeting USP7 further suppresses the in vitro proliferation of USP22-knockout (USP22-Ko) A549 and H1299 lung cancer cells and induces a stronger activation of p53 tumor suppressor signaling pathway. In addition, USP22-Ko cancer cells are more sensitive to a combination of cisplatin and USP7 inhibitor. USP7 inhibitor treatment further suppresses in vivo angiogenesis and tumor growth and induced more apoptosis in USP22-Ko cancer xenografts. Taken together, our findings demonstrate that USP7 inhibition can dramatically upregulate USP22 in cancer cells; and targeting USP7 and USP22 may represent a more effective approach for targeted cancer therapy, which warrants further study. Video Abstract.

Efficacy and safety of PD-1 inhibitor combined with concurrent chemoradiotherapy in locally advanced cervical cancer with pelvic and/or para-aortic lymph node metastases: a retrospective cohort study.

BACKGROUND: The prognosis remains poor after standard chemoradiotherapy in locally advanced cervical cancer patients with pelvic and/or para-aortic lymph node metastases. Programmed cell death receptor-1 (PD-1) inhibitors have been recommended as the first-line treatment for recurrent cervical cancer. The efficacy of PD-1 inhibitor combined with concurrent chemoradiotherapy in locally advanced cervical cancer was still uncertain. This study aimed to explore the efficacy and safety of PD-1 inhibitors combined with concurrent chemoradiotherapy in locally advanced cervical cancer patients with pelvic and/or para-aortic lymph node metastases. METHODS: This retrospective study included patients with pelvic and/or para-aortic lymph node positive diseases [International Federation of Gynecology and Obstetrics (FIGO) stage IIB-IVA] who had received PD-1 inhibitors plus chemoradiotherapy/radiotherapy between April 1, 2020, and March 31, 2022 at the Hunan Cancer Hospital. The baseline clinicopathological characteristics, treatment, and clinical outcomes were collected. The major clinical outcomes were objective response rate (ORR), progression-free survival (PFS), and treatment-related adverse events (TRAEs). RESULTS: A total of 29 patients were included. The mean age was 55.8 [standard deviation (SD): 8.8] years. Most patients had stage IIIA-IIIB disease (72.4%) and squamous cell carcinoma (93.1%). All patients had lymph node metastases, including 24 (82.8%) with multiple metastases and 11 (37.9%) with para-aortic lymph node metastases. Among the 29 patients, 18 received sintilimab and 11 received camrelizumab concurrently with chemoradiotherapy or radiotherapy. The ORR was 96.6% [95% confidence interval (CI): 0.828, 0.993] at 3 months after radiotherapy (including 15 complete responses and 13 partial responses). At the data cutoff (August 31, 2022), the median follow-up was 14 (range, 5-30) months. The median PFS was not mature. The estimated 1- and 2-year PFS rates were 85.3% (95% CI: 60.1%, 95.2%) and 76.8% (95% CI: 47.0%, 91.2%), respectively. TRAEs of any grade occurred in 27 (93.1%) patients, most commonly as a decrease in white blood counts (82.8%), anemia (58.6%), and fatigue (48.3%). TRAEs of grade 3 or greater occurred in eight (27.6%) patients. There were no treatment-related deaths. CONCLUSIONS: PD-1 inhibitor combined with concurrent chemoradiotherapy showed potential benefit in term of tumor response and PFS in locally advanced cervical cancer patients with pelvic and/or para-aortic lymph node metastases.

A phase 1 trial of fuzuloparib in combination with apatinib for advanced ovarian and triple-negative breast cancer: efficacy, safety, pharmacokinetics and germline BRCA mutation analysis.

BACKGROUND: The effect of the combination of an anti-angiogenic agent with a poly (ADP-ribose) polymerase (PARP) inhibitor in cancer treatment is unclear. We assessed the oral combination of fuzuloparib, a PARP inhibitor, and apatinib, a VEGFR2 inhibitor for treating advanced ovarian cancer (OC) or triple-negative breast cancer (TNBC). METHODS: This dose-escalation and pharmacokinetics-expansion phase 1 trial was conducted in China. We used a standard 3 + 3 dose-escalation design, with 7 dose levels tested. Patients received fuzuloparib orally twice daily, and apatinib orally once daily. The study objectives were to determine the safety profile, recommended phase 2 dose (RP2D), pharmacokinetics, preliminary efficacy, and efficacy in relation to germline BRCA mutation (gBRCAmut). RESULTS: Fifty-two pre-treated patients were enrolled (30 OC/22 TNBC). 5 (9.6%) patients had complete response, 14 (26.9%) had partial response, and 15 (28.8%) had stable disease. Objective response rate (ORR) and disease control rate were 36.5% (95% CI 23.6-51.0) and 65.4% (95% CI 50.9-78.0), respectively. At the highest dose level of fuzuloparib 100 mg plus apatinib 500 mg, the ORR was 50.0% (4/8; 95% CI 15.7-84.3); this dose was determined to be the RP2D. Patients with gBRCAmut had higher ORR and longer median progression-free survival (PFS) than those with gBRCAwt, both in OC (ORR, 62.5% [5/8] vs 40.9% [9/22]; PFS, 9.4 vs 6.7 months) and TNBC (ORR, 66.7% [2/3] vs 15.8% [3/19]; PFS, 5.6 vs 2.8 months). Two dose-limiting toxicities occurred: grade 4 febrile neutropenia (fuzuloparib 100 mg plus apatinib 250 mg) and thrombocytopenia (fuzuloparib 100 mg plus apatinib 375 mg). Maximum tolerated dose was not reached. The most common treatment-related grade ≥ 3 toxicities in all patients were hypertension (19.2%), anaemia (13.5%), and decreased platelet count (5.8%). Exposure of apatinib increased proportionally with increasing dose ranging from 250 to 500 mg, when combined with fuzuloparib 100 mg. CONCLUSIONS: Fuzuloparib plus apatinib had acceptable safety in patients with advanced OC or TNBC. Fuzuloparib 100 mg bid plus apatinib 500 mg qd was established as the RP2D. With the promising clinical activity observed, this combination is warranted to be further explored as a potential alternative to chemotherapy. TRIAL REGISTRATION: ClinicalTrials.gov, NCT03075462 (Mar. 9, 2017).

Adsorption properties and mechanism research of phosphorus with different molecular structures from aqueous solutions by La-modified biochar.

In order to explore the adsorption characteristics of phosphorus from molecules with different molecular structures and varying number of phosphate groups on metal-modified biochar, walnut shell biochar was modified with LaCl3 to prepare lanthanum-loaded biochar (BC-La). Adsorption of four polar components, namely phytic acid (IHP), adenosine-5'-disodium triphosphate (5-ATP), hydroxyethylidene diphosphonic acid (HEDP), and sodium pyrophosphate (PP), was studied. The adsorption properties and mechanism of phosphorus sorption by BC-La were analyzed by SEM-EDS and FTIR for the different structures. The results showed that the maximum adsorption capacity of BC-La for IHP, 5-ATP, HEDP, and PP was 85.85, 9.04, 15.80, and 14.45 mg/g, respectively. The adsorption capacity was positively correlated with the polarity of organic phosphorus. The adsorption behavior conformed to the quasi second-order kinetic fitting equation, and the increase of temperature was conducive to the removal of all four phosphorus pollutants. BC-La adsorbs IHP and HEDP mainly through electrostatic attraction. The adsorption of 5-ATP and PP is dominated by complexation. The La-modified biochar has broad prospects in water remediation, which can provide a theoretical basis for removal of different forms of phosphorus pollutants and prevention and control of water eutrophication.

ARL4C depletion suppresses the resistance of ovarian cancer to carboplatin by disrupting cholesterol transport and autophagy via notch-RBP-Jκ-H3K4Me3-OSBPL5.

Increasing studies indicate that cholesterol plays an important role in drug resistance. ARL4C is implicated in the export and import of cholesterol, therefore this study aimed to explore the effect of ARL4C on the resistance of ovarian cancer (OVC) to Carboplatin. This study collected OVC tissue samples from patients who are sensitive or resistant to carboplatin, and established Carboplatin-resistant OVC cell lines, OVCAR3(R) and SKOV3(R) using OVCAR3 and SKOV3. High throughput sequencing was conducted to find genes that regulated by ARL4C. Cholesterol esterification was performed to evaluate the transport of cholesterol from Lysosome (LY) to Endoplasmic reticulum (ER). The fluorescence of LC3-GFP-mRFP was used to evaluate the function of autophagy flux. As indicated by PCR, western blot and Immunohistochemistry, ARL4C was increased in the Carboplatin-resistant OVC tissues and cells. Knockdown of ARL4C attenuated the resistance of OVCAR3(R) and SKOV3(R) to Carboplatin. By suppressing Notch signal, ARL4C knockdown inhibited the transcriptional function of RBP-Jκ and RBP-Jκ-induced H3K4Me3, which collectively reduced OSBPL5 expression. OSBPL5 deficiency inhibited the transport of cholesterol from LYs to ER, which led to the accumulation of cholesterol in LYs and the dysfunction of autophagy. In summary, ARL4C knockdown attenuated the resistance of OVC to Carboplatin by disrupting cholesterol transport and autophagy. This study revealed a promising target to attenuate the resistance of OVC to Carboplatin and elucidated the potential mechanism.

LncRNA LAMTOR5-AS1 sponges miR-210-3p and regulates cervical cancer progression.

AIM: Cervical cancer has attracted increasing attention in recent years, and the incidence has shown a trend of younger age. Therefore, it is an effective method to regulate the progression of cervical cancer through new prognostic biomarkers. The purpose of this study was to evaluate the potential of lncRNA LAMTOR5-AS1 (LAMTOR5-AS1) as a prognostic biomarker and reveal its regulatory role in cervical cancer. METHODS: A total of 120 patients with cervical cancer were selected as research subjects to verify the prognostic effect of LAMTOR5-AS1 in a series of experiments. The expression of LAMTOR5-AS1 in cervical cancer tissues and cells was determined by polymerase chain reaction assay. The proliferation, migration, and invasion ability of cervical cancer cells were evaluated by Cell Counting Kit-8 (CCK-8) and Transwell assay. Luciferase reporter gene detection was used to determine the mechanism of LAMTOR5-AS1 targeting miR-210-3p, and to reflect the prognostic value of LAMTOR5-AS1 according to statistical methods. RESULTS: LAMTOR5-AS1 decreased in cervical cancer tissues, while miR-210-3p expression increased. In the study of cervical cancer cells, it was found that the LAMTOR5-AS1 sponge miR-210-3p was associated with the malignant progression of cervical cancer. Overexpression of LAMTOR5-AS1 could effectively inhibit the development of cervical cancer cells and might be chosen as a prognostic biomarker of cervical cancer. CONCLUSIONS: LAMTOR5-AS1 sponges miR-210-3p and modulates the progression of cervical cancer, which predict the prognosis of cervical cancer patients.

Transcription factor AP2 enhances malignancy of non-small cell lung cancer through upregulation of USP22 gene expression.

BACKGROUND: Ubiquitin-specific protease 22 (USP22), a putative cancer stem cell marker, is frequently upregulated in cancers, and USP22 overexpression is associated with aggressive growth, metastasis, and therapy resistance in various human cancers including lung cancer. However, USP22 gene amplification seldom occurs, and the mechanism underlying USP22 upregulation in human cancers remains largely unknown. METHODS: A luciferase reporter driven by a promoter region of USP22 gene was selectively constructed to screen against a customized siRNA library targeting 89 selected transcription factors to identify potential transcription factors (TFs) that regulate USP22 expression in human non-small cell lung cancers (NSCLC). Association of identified TFs with USP22 and potential role of the TFs were validated and explored in NSCLC by biological assays and immunohistochemistry analysis. RESULTS: Luciferase reporter assays revealed that SP1 and activating transcription factor 3 (ATF3) inhibit USP22 transcription, while transcription factor AP-2 Alpha/Beta (TFAP2A/2B) and c-Myc promote USP22 transcription. Binding site-directed mutagenesis and chromosome immunoprecipitation (ChIP) assays validated AP2α and AP2ß are novel TFs of USP22. Furthermore, overexpression of AP2A and AP2B significantly upregulates USP22 expression, and its target: Cyclin D1, concurrently enhances the proliferation, migration, and invasion of NSCLC A549 and H1299 cells in a partially USP22-dependent manner. Moreover, AP2 protein level correlated with USP22 protein in human NSCLC tissues. CONCLUSION: Our findings indicate AP2α and AP2ß are important transcription factors driving USP22 gene expression to promote the progression of NSCLC, and further support USP22 as a potential biomarker and therapeutic target for lung cancer. Video Abstract.

An exhaustive investigation on antibiotics contamination from livestock farms within sensitive reservoir water area: Spatial density, source apportionment and risk assessment.

Although the studies on antibiotic contamination are common at present, large-scale sampling studies drawing highly representative conclusions are still scarce. This study conducted a comprehensive investigation on a total of 1183 samples from 70 livestock farms within a sensitive area around reservoir waters. 45 types of antibiotics belonging to 5 different classes were monitored. This is the first analysis to comprehensively investigate the density distribution, source apportionment, ecological and health risk of antibiotics in an entire area of sensitive waters. The results showed that the layer manure samples had highest detection rate of antibiotics (0.0 %-96.1 %, average value = 30.7 %) followed by pig manure samples. Oxytetracycline had the highest concentration of 712.16 mg/kg in a pig manure sample. Different from using antibiotic concentration as a proxy for pollution level, the spatial density was calculated by averaging antibiotic concentration to area and converting different livestock to pig equivalent. The spatial density of pig equivalent can more realistically reflect the pollution caused by different breeds of livestocks. It was shown that the pig farms contributed higher to total antibiotic density than the layer and cattle farms did. After assessed, a few antibiotics (oxytetracycline, chlorotetracycline and tetracycline) have posed high ecological risks to soil around the farms. However, none of them caused hazard quotient (HQ) risk and carcinogenic risk (CR) to human health in the water of reservoir. Children were more likely to be at hazard risk than adults. Antibiotic mass fluctuation rules were analyzed along the chain (feed → livestock waste → soil → surface water). Feed, livestock waste and soil had similar diversity, but the antibiotic concentrations continued to decline, implying the possible sources of antibiotic residues were similar. Thus, it is important to reduce unnecessary antibiotic use to prevent the potential long-term risk of antibiotics.

Oyster Shell Modified Tobacco Straw Biochar: Efficient Phosphate Adsorption at Wide Range of pH Values.

In order to improve the phosphate adsorption capacity of Ca-loaded biochar at a wide range of pH values, Ca (oyster shell) was loaded as Ca(OH)2 on the tobacco stalk biochar (Ca-BC), which was prepared by high-temperature calcination, ultrasonic treatment, and stirring impregnation method. The phosphorus removal performance of Ca-BC adsorption was studied by batch adsorption experiments, and the mechanism of Ca-BC adsorption and phosphorus removal was investigated by SEM-EDS, FTIR, and XRD. The results showed that after high-temperature calcination, oyster shells became CaO, then converted into Ca(OH)2 in the process of stirring impregnation and had activated the pore expansion effect of biochar. According to the Langmuir model, the adsorption capacity of Ca-BC for phosphate was 88.64 mg P/g, and the adsorption process followed pseudo-second-order kinetics. The Ca(OH)2 on the surface of biochar under the initial pH acidic condition preferentially neutralizes with H+ acid-base in solution, so that Ca-BC chemically precipitates with phosphate under alkaline conditions, which increases the adsorption capacity by 3-15 times compared with other Ca-loaded biochar. Ca-BC phosphate removal rate of livestock wastewater (pig and cattle farms) is 91~95%, whereas pond and domestic wastewater can be quantitatively removed. This study provides an experimental basis for efficient phosphorus removal by Ca-modified biochar and suggesting possible applications in real wastewater.

Effectiveness of layered inoculation in solid-state anaerobic co-digestion of pig manure and corn straw: Focus on macro-, micro-, and genetic-levels.

Layered inoculation can achieve rapid start-up and promote methanation performance of anaerobic digesters. Daily specific methane yield (SMY) rapidly increased to 2.93 mL/g VS/d during 0-13 days, and cumulative SMY reached 212 mL/g VS in the solid-state anaerobic co-digestion (SS-AcoD) of pig manure and corn straw. Data were collected at macro-, micro-, and genetic-levels of each substrate layer. The results showed that layered inoculation could improve volatile fatty acids utilization and prevent adverse effects of high total ammonium nitrogen concentrations. Layered inoculation accelerated hydrolysis, acidification, and methanogenesis of substrates, as evidenced by the efficient inoculation of Bacteroidetes, Anaerolineales, Methanosphaerula, and Methanothrix, which were primarily from inocula. The various stages of SS-AcoD were synergistically initiated during the first 13 days, and acetoclastic pathway was boosted. These results further explain why layered inoculation is an efficient method for improving methanation performance of SS-AcoD and achieving efficient utilization of organic solid waste.

Determination of 38 antibiotics in raw and treated wastewater from swine farms using liquid chromatography-mass spectrometry.

The present study firstly aimed at developing a multi-residue method to identify and quantify 38 veterinary antibiotics (belonging to five different classes) not only for raw swine wastewater but also for wastewater differently treated by different units. The proposed method is based on a solid-phase extraction procedure and ultra high performance liquid chromatography with mass spectrometry. For sample preparation, the optimal loading sample volume was selected as 50 mL, the pH of which was adjusted to approximately 3.0 using formic acid. Then 0.1 g/L ethylenediaminetetraacetic acid disodium salt was added. The recovery rates for different types of wastewaters were in the range of 35.94-124.51% and the relative standard deviations were in the range of 0.36-14.62%. All the matrix standard curves exhibited high linearity (0.9956-0.9999). The matrix effects for the target antibiotics ranged from -61.73 to +148.75%. To ensure the practicality of the method, we performed the detection of the actually added concentration to determine method detection limits and quantitation limits. The quantitation limits of most of the target antibiotics were 0.04 µg/L, except for spiramycin (0.1 µg/L) and roxithromycin (0.2 µg/L). This optimized and validated method was applied to analyze antibiotic residues in swine water samples from four swine farms.

Diagnostic accuracy of convolutional neural network-based endoscopic image analysis in diagnosing gastric cancer and predicting its invasion depth: a systematic review and meta-analysis.

BACKGROUND AND AIMS: This study aimed to evaluate the accuracy and effectiveness of the convolutional neural network (CNN) in diagnosing gastric cancer and predicting the invasion depth of gastric cancer and to compare the performance of the CNN with that of endoscopists. METHODS: PubMed, Embase, Web of Science, and gray literature were searched until July 23, 2021 for studies that assessed the diagnostic accuracy of CNN-assisted examinations for gastric cancer or the invasion depth of gastric cancer. Studies meeting inclusion criteria were included in the systematic review and meta-analysis. RESULTS: Seventeen studies comprising 51,446 images and 174 videos of 5539 patients were included. The pooled sensitivity, specificity, positive likelihood ratio (LR+), negative likelihood ratio (LR-), and area under the curve (AUC) of the CNN for diagnosing gastric cancer were 89% (95% confidence interval [CI], 85-93), 93% (95% CI, 88-97), 13.4 (95% CI, 7.3-25.5), .11 (95% CI, .07-.17), and .94 (95% CI, .91-.98), respectively. The performance of the CNN in diagnosing gastric cancer was not significantly different from that of expert endoscopists (.95 vs .90, P > .05) and was better than that of overall endoscopists (experts and nonexperts) (.95 vs .87, P < .05). The pooled sensitivity, specificity, LR+, LR-, and AUC of the CNN for predicting the invasion depth of gastric cancer were 82% (95% CI, 78-85), 90% (95% CI, 82-95), 8.4 (95% CI, 4.2-16.8), .20 (95% CI, .16-.26), and .90 (95% CI, .87-.93), respectively. CONCLUSIONS: The CNN is highly accurate in diagnosing gastric cancer and predicting the invasion depth of gastric cancer. The performance of the CNN in diagnosing gastric cancer is not significantly different from that of expert endoscopists. Studies of the real-time performance of the CNN for gastric cancer diagnosis are needed to confirm these findings.

Preferred method of therapy for patients with early-stage high-grade neuroendocrine carcinoma of the cervix.

High-grade neuroendocrine carcinoma of the uterine cervix (HGNECC) is a rare and overly aggressive malignancy. Due to its rarity, there is no standard treatment. A majority of early-stage patients receive radical hysterectomy and lymph node dissection (RH+LND), followed by adjuvant chemotherapy. To explore the most suitable methods of therapy, a multicenter retrospective review of HGNECC patients was conducted. A total of 133 patients (I-IIA, FIGO 2009) treated from March 2003 to September 2018 were enrolled in this study. The 5-year DFS and OS rates for stages IB and IIA were 44.8% and 39.5%, and 53.8% and 39.6%, respectively. The median DFS and OS for stages IB and IIA were 41 months and 12 months, and 63 months and 45 months, respectively. The RH+LND surgery procedure was associated with a significantly better DFS (P=0.015) and OS (P=0.006), while the bilateral salpingo-oophorectomy (BSOE) was also associated with a better OS (P=0.023). The efficacy of paclitaxel-platinum (TP/C) adjuvant chemotherapy regimens need to be confirmed using clinical trials, especially for tumors with a diameter of >4 cm (P=0.0005). Therefore, the RH+LND+BSOE procedure was recommended for HGNECC patients at stages IB-IIA. TP/C is an alternative chemotherapy regimen that results in optimal survival. Moreover, a tumor diameter of >4 cm, LNM, DSI, and LVSI were confirmed as high-risk factors for worse DFS and OS. Patients without risk factor, 1 or 2 or 3 risk factors, and 4 risk factors had significantly different DFS and OS values.

LncRNA RP11-499E18.1 Inhibits Proliferation, Migration, and Epithelial-Mesenchymal Transition Process of Ovarian Cancer Cells by Dissociating PAK2-SOX2 Interaction.

Background: Ovarian cancer (OC)is a deadly gynecological malignancy worldwide. It is urgent to identify diagnostic biomarkers of OC to disclose the underlying mechanism. Methods and Materials: Bioinformatics analysis was used to identify target genes. Gene expression was detected and altered by qRT-PCR and cell transfection, respectively. The interaction between RP11-499E18.1 and PAK2, as well as that between PAK2 and SOX2, was determined using RNA pulldown, RNA immunoprecipitation (RIP), and co-immunoprecipitation (co-IP) assay, respectively. Localizations of RP11-499E18.1, PAK2, and SOX2 were respectively determined employing immunohistochemical (IHC), IF, and FISH. The regulatory effects of RP11-499E18.1, PAK2, and SOX2 on OC cell proliferation, migration, colony formation, epithelial-mesenchymal transition (EMT)-related factor expression, and SOX2 nuclear translocation were determined. Finally, the effects of RP11-499E18.1 and PAK2 expression on the tumor growth in nude mice were determined. Results: RP11-499E18.1, PAK2, and SOX2 were selected in our study. RP11-499E18.1 was downregulated, while PAK2 and SOX2 was upregulated in OC tissues and cells. RP11-499E18.1 coexists in the nucleus and cytoplasm of OC cells. There is an interaction between RP11-499E18.1 and PAK2, as well as PAK2 and SOX2 in OC cells. Alteration of RP11-499E18.1 and PAK2 expression both had no influence on PAK2 and SOX2 levels, but PAK2 upregulation notably augmented p-SOX2 level. RP11-499E18.1 overexpression suppressed OC cell proliferation, migration, and colony formation, as well as SOX2 nuclear translocation. Besides, it inhibited tumor growth in nude mice. However, these effects were notably reversed by PAK2 upregulation and eventually offset by SOX2 knockdown. Additionally, RP11-499E18.1 overexpression reduced PAK2-SOX2 interaction and SOX phosphorylation, and increased the binding of RP11-499E18.1 by PAK2. Conclusion: These lines of evidence demonstrated that RP11-499E18.1 might play its tumor suppressor roles in OC via regulation of the RP11-499E18.1-PAK2-SOX2 axis. This research indicated that RP11-499E18.1 might be used as a diagnostic biomarker for OC in the future.

Rs3802278 in 3'-UTR of SULF1 associated with platinum resistance and survival in Chinese epithelial ovarian cancer patients.

Ovarian cancer is the leading cause of death from gynecologic cancers, but platinum resistance remains a major obstacle in the chemotherapy of ovarian cancer. This study aims to examine the role of polymorphisms in sulfatase 1 (SULF1) in platinum resistance and survival in advanced epithelial ovarian cancer (EOC) patients. We genotyped 12 SNPs of SULF1 in 195 EOC patients treated with platinum using MassARRAY method and evaluated the association between the SNPs and platinum response. SULF1 rs3802278 was marginal significantly associated with platinum resistance in recessive model with p value of 0.055. The patients with SULF1 rs3802278 AA were more resistant to platinum-based chemotherapy comparing to those with AG/GG genotype (OR: 2.317, 95%CI: 0.982 ∼ 5.465). In survival analysis, rs3802278 was significantly associated with both of PFS and OS after adjusted by FIGO stage and age. Patients with AA genotypes showed a shorter PFS and OS than with AG/GG genotypes (median PFS: 15 months vs. 21 months, p = 0.010, HR = 1.876, 95%CI: 1.165-3.022; median OS: 42 months vs. 73 months, p = 0.031, HR = 1.928, 95%CI: 1.061-3.504). SULF1 rs3802278 may serve as a potential candidate biomarker for the prediction of platinum resistance and prognosis in Chinese EOC patients.

G9a Promotes Invasion and Metastasis of Non-Small Cell Lung Cancer through Enhancing Focal Adhesion Kinase Activation via NF-κB Signaling Pathway.

Potential roles of euchromatic histone methyltransferase 2 (EHMT2 or G9a) in invasion and metastasis are not well understood in non-small cell lung cancer (NSCLC). Here, we investigated the effect and underlying mechanisms of G9a and therapeutic implications of targeting G9a in the invasion and metastasis of NSCLC. Overexpression of G9a significantly enhanced in vitro proliferation and invasion, while knockdown of G9a drastically suppressed in vivo growth and metastasis of A549 and H1299 NSCLC cells. Knockdown or inhibition of G9a significantly decreased the expression of focal adhesion kinase (FAK) protein and activation of FAK pathway. In addition, defactinib, a potent FAK inhibitor, partially abolished the G9a-enhanced invasion in these NSCLC cells. Furthermore, targeting G9a was found to suppress NF-κB transcriptional activity in NSCLC cells through stabilizing NF-κB inhibitor alpha (IκBα), while an NF-κB inhibitor Parthenilide partially abolished the G9a-enhanced FAK activation, which suggests that G9a-enhanced invasion and activation of FAK is mediated by elevated NF-κB activity. Notably, a strong positive correlation between the IHC staining of G9a and phosphorylated FAK proteins was identified in H1299 xenografts and 159 cases of NSCLC tissues (R = 0.408). IMPLICATIONS: The findings of this study strongly demonstrate that G9a may promote invasion and metastasis of NSCLC cells by enhancing FAK signaling pathway via elevating NF-κB transcriptional activity, indicating potential significance and therapeutic implications of these pathways in the invasion and metastasis of NSCLCs that overexpress G9a protein.