Microbial ß-Glucuronidase Hydrogel Beads Activate Chemotherapeutic Prodrug.

Bacteria-assisted chemotherapeutics have been highlighted as an alternative or supplementary approach to treating cancer. However, dynamic cancer-microbe studies at the in vitro level have remained a challenge to show the impact and effectiveness of microbial therapeutics due to the lack of relevant coculture models. Here, we demonstrate a hydrogel-based compartmentalized system for prodrug activation of a natural ingredient of licorice root, glycyrrhizin, by microbial ß-glucuronidase (GUS). Hydrogel containment with Lactococcus lactis provides a favorable niche to encode GUS enzymes with excellent permeability and can serve as an independent ecosystem in the transformation of pro-apoptotic materials. Based on the confinement system of GUS expressing microbes, we quantitatively evaluated chemotherapeutic effects enhanced by microbial GUS enzyme in two dynamic coculture models in vitro (i.e., 2D monolayered cancer cells and 3D tumor spheroids). Our findings support the processes of prodrug conversion mediated by bacterial GUS enzyme which can enhance the therapeutic efficacy of a chemotherapy drug under dynamic coculture conditions. We expect our in vitro coculture platforms can be used for the evaluation of pharmacological properties and biological activity of xenobiotics as well as the potential impact of microbes on cancer therapeutics.

The 8th AJCC classification is inferior to a new neck stage based on intraparotid lymph node in parotid gland cancer.

BACKGROUND: Lymph node (LN) status is an important prognostic factor for parotid gland cancer (PGC). This study aimed to analyze the impact of extranodal extension (ENE) of intraparotid LN and LN metastasis burden on survival in PGC. METHODS: Patients with surgically treated PGC and at least one metastatic cervical LN were retrospectively enrolled. Primary outcome variables were distant metastasis-free survival (DMFS), disease-specific survival (DSS), and overall survival (OS). The impact of ENE and LN metastasis burden was assessed using the Cox model. RESULTS: A total of 292 patients were included. ENE in cervical or intraparotid LN was not associated with DMFS, DSS, or OS. Intraparotid LN metastasis had a significant impact on prognosis, and the presence of only one metastatic intraparotid LN offered an approximately 1.5-fold risk of distant metastasis. Prognostic models based on the number of positive LNs (1 vs. 2-3 vs. 4+) were superior to the AJCC N stage in terms of DMFS, DSS, and OS. CONCLUSIONS: ENE of cervical or intraparotid LN has a limited effect on the prognosis of PGC, and the number of positive LNs is better than the AJCC N stage in LN status evaluation.

Overexpressed Trophoblast Glycoprotein Contributes to Preeclampsia Development by Inducing Abnormal Trophoblast Migration and Invasion Toward the Uterine Spiral Artery.

BACKGROUND: Preeclampsia is a significant pregnancy disorder with an unknown cause, mainly attributed to impaired spiral arterial remodeling. METHODS: Using RNA sequencing, we identified key genes in placental tissues from healthy individuals and preeclampsia patients. Placenta and plasma samples from pregnant women were collected to detect the expression of TPBG (trophoblast glycoprotein). Pregnant rats were injected with TPBG-carrying adenovirus to detect preeclamptic features. HTR-8/SVneo cells transfected with a TPBG overexpression lentiviral vector were used in cell function experiments. The downstream molecular mechanisms of TPBG were explored using RNA sequencing and single-cell RNA sequencing data. TPBG expression was knocked down in the lipopolysaccharide-induced preeclampsia-like rat model to rescue the preeclampsia features. We also assessed TPBG's potential as an early preeclampsia predictor using clinical plasma samples. RESULTS: TPBG emerged as a crucial differentially expressed gene, expressed specifically in syncytiotrophoblasts and extravillous trophoblasts. Subsequently, we established a rat model with preeclampsia-like phenotypes by intravenously injecting TPBG-expressing adenoviruses, observing impaired spiral arterial remodeling, thus indicating a causal correlation between TPBG overexpression and preeclampsia. Studies with HTR-8/SVneo cells, chorionic villous explants, and transwell assays showed TPBG overexpression disrupts trophoblast/extravillous trophoblast migration/invasion and chemotaxis. Notably, TPBG knockdown alleviated the lipopolysaccharide-induced preeclampsia-like rat model. We enhanced preeclampsia risk prediction in early gestation by combining TPBG expression with established clinical predictors. CONCLUSIONS: These findings are the first to show that TPBG overexpression contributes to preeclampsia development by affecting uterine spiral artery remodeling. We propose TPBG levels in maternal blood as a predictor of preeclampsia risk. The proposed mechanism by which TPBG overexpression contributes to the occurrence of preeclampsia via its disruptive effect on trophoblast and extravillous trophoblast migration/invasion on uterine spiral artery remodeling, thereby increasing the risk of preeclampsia.

Nano oxygen chamber by cascade reaction for hypoxia mitigation and reactive oxygen species scavenging in wound healing.

Hypoxia, excessive reactive oxygen species (ROS), and impaired angiogenesis are prominent obstacles to wound healing following trauma and surgical procedures, often leading to the development of keloids and hypertrophic scars. To address these challenges, a novel approach has been proposed, involving the development of a cascade enzymatic reaction-based nanocarriers-laden wound dressing. This advanced technology incorporates superoxide dismutase modified oxygen nanobubbles and catalase modified oxygen nanobubbles within an alginate hydrogel matrix. The oxygen nano chamber functions through a cascade reaction between superoxide dismutase and catalase, wherein excessive superoxide in the wound environment is enzymatically decomposed into hydrogen peroxide, and this hydrogen peroxide is subsequently converted into oxygen by catalase. This enzymatic cascade effectively controls wound inflammation and hypoxia, mitigating the risk of keloid formation. Concurrently, the oxygen nanobubbles release oxygen continuously, thus providing a sustained supply of oxygen to the wound site. The oxygen release from this dynamic system stimulates fibroblast proliferation, fosters the formation of new blood vessels, and contributes to the overall wound healing process. In the rat full-thickness wound model, the cascade reaction-based nano oxygen chamber displayed a notable capacity to expedite wound healing without scarring. Furthermore, in the pilot study of porcine full-thickness wound healing, a notable acceleration of tissue repair was observed in the conceived cascade reaction-based gel treated group within the 3 days post-surgery, which represents the proliferation stage of healing process. These achievements hold significant importance in ensuring the complete functional recovery of tissues, thereby highlighting its potential as a promising approach for enhancing wound healing outcomes.

Intraparotid lymph node metastasis affects distant metastasis in parotid adenoid cystic carcinoma.

To evaluate the relationship between factors of metastatic intraparotid lymph node (IPLN) and distant metastasis in parotid adenoid cystic carcinoma (ACC). Patients with surgically treated parotid ACC were retrospectively enrolled, and primary outcome variable was distant metastasis free survival (DMFS). The effect of factors of metastatic IPLN on DMFS was evaluated using Cox model. In total, 232 patients were included. Extranodal extension of IPLN and cervical lymph nodes did not impact the DMFS, and the 7th but not 8th AJCC N stage was associated with DMFS. Groups of 0 and 1 metastatic IPLN had comparable DMFS, but presence of 2+ positive IPLN was related to increased worse DMFS (p = 0.034, HR 2.09). A new N stage (0 vs 1-2 vs 3+) based on total positive lymph node number exhibited better C-index than traditional N stage. IPLN metastasis increased the risk of distant metastasis, and the impact was mainly determined by the number of metastatic IPLN. Our proposed N stage provided better DMFS prediction than the 8th AJCC N classification.

Hyperoside suppresses osteoclasts differentiation and function through downregulating TRAF6/p38 MAPK signaling pathway.

Hyperoside (HP), as a natural product, can promote proliferation and differentiation of osteoblasts and presents a protective effect on ovariectomized (OVX) mice. However, the inhibitory effect of HP on osteoclasts (OCs) and the potential mechanism remain to be elucidated. In this study, it was found that HP could effectively inhibit the differentiation and bone resorption of OCs, and its intrinsic molecular mechanism was related to the inhibition of TRAF6/p38 MAPK signaling pathway. Therefore, HP could be a promising natural compound for lytic bone diseases.

Comparison of Clinical Results and Quality-of-Life in Tongue Cancer Patients Undergoing Submental Island Flap and Radial Forearm Free Flap Reconstruction.

PURPOSE: Our goal was to compare the clinical results and quality-of-life (QoL) in tongue cancer patients undergoing submental island pedicled (SIP) flap and radial forearm free (RFF) flap reconstruction. MATERIALS AND METHODS: Patients undergoing SIP or RFF flap reconstruction for primary tongue squamous cell carcinoma were prospectively enrolled and were asked to complete the University of Washington Quality-of-Life (UW-QOL) questionnaire, version 4, at 12 months after the operation. The study's main interest was QoL as well as locoregional recurrence control. RESULTS: A total of 190 patients were enrolled for analysis, and the SIP and RFF groups showed significant differences in patient age, American Society of Anesthesiologists classification, and hospital cost. In the survival analysis, locoregional recurrence occurred in 35 patients in the SIP group and 48 patients in the RFF group; the difference was not significant (P = .440). In the QoL analysis, compared with patients in the SIP group, those in the RFF group had higher scores in the domains of activity and recreation. No significant differences were found with respect to the other domains. CONCLUSIONS: The SIP flap and RFF flap have comparable survival control in tongue squamous cell carcinoma patients. The RFF flap might lead to better QoL, but the SIP flap imposes fewer limitations on patients' health status and is associated with lower hospital cost.

Oxygen therapy alleviates hepatic steatosis by inhibiting hypoxia-inducible factor-2α.

Non-alcoholic fatty liver disease (NAFLD) is difficult to manage due to the lack of effective treatments. Increased oxygen consumption caused by overnutrition, along with reduced oxygen delivery to liver cells induces hepatic steatosis. Here, we investigated the efficacy of oxygen therapy (OT) to alleviate hepatic steatosis. The effect of OT on hepatic steatosis was evaluated in high-fat-diet (HFD)-fed mice and palmitic acid (PA)-treated primary hepatocytes. Liver biopsy tissue samples were used to determine the relationship between the expression of hypoxia-inducible factor-2α (HIF-2α) and the progression of NAFLD. The role of HIF-2α in the OT group was determined based on the overexpression of HIF-2α in vitro. OT safely alleviated hepatic hypoxia and improved hepatic steatosis by inhibiting hepatic de novo lipogenesis in HFD-fed mice and PA-treated primary hepatocytes, and this was accompanied by reduced expression of HIF-2α and hepatic de novo lipogenesis. The analysis of liver tissues from individuals with or without NAFLD revealed a positive correlation between hepatic HIF-2α expression and NAFLD progression. Overexpression of HIF-2α in vitro inhibited the beneficial effect of OT against hepatic lipogenesis and steatosis. OT might be a viable treatment option for NAFLD and functions by alleviating hypoxia and inhibiting the liver HIF-2α signaling pathway.

An Oxygen Self-Evolving, Multistage Delivery System for Deeply Located Hypoxic Tumor Treatment.

The hypoxia-induced resistance to radiotherapy (RT) is a great threat to cancer patients. Therefore, overcoming the hypoxia tumor microenvironment is a vital issue. Herein, spindle-shaped CuS@CeO2 core-shell nanoparticles combining self-supplied oxygen, photothermal ability, and RT sensitive to cancer therapy are introduced. The spindle shape of CuS@CeO2 core-shell nanoparticles can potentiate their tumor penetration and subsequent internalization by cancer cells. The presence of CeO2 , functioning as a nanoenzyme, catalyzes the endogenous H2 O2 in tumor tissue into O2 , which remodels the hypoxic microenvironment into one susceptible to RT. CuS nanoparticles encapsulated in CeO2 undergo a steady release and deep tumor penetration, allowing the regression of lesions less affected by RT. Furthermore, in vitro and in vivo studies reveal that the design not only mitigates the dosage of RT, but more importantly allows the entire tumor to be treated without relapses.

Management and prognosis of cancers in the accessory parotid gland.

OBJECTIVE: This study was performed to analyze the clinical management of accessory parotid gland (APG) cancer and possible risk factors for disease-related death. METHODS: Patients diagnosed with primary APG cancers in the largest medical center in Northeast China were enrolled from January 1990 to December 2016. RESULTS: All 43 patients underwent resection of the tumors and superficial parotid gland by a standard Blair incision. Seven (16.3%) patients also required selective neck dissection. The most common lesion was mucoepidermoid carcinoma. Temporary facial paralysis occurred in 11 (25.6%) patients, and permanent facial paralysis occurred in 3 (7.0%) patients because of surgical resection of the facial nerve, which was involved with the tumor. The 5- and 10-year disease-specific survival rates were 86.0% and 66.0%, respectively. The tumor stage, neck status, neck dissection, and tumor grade were significantly associated with disease-related death, but only the tumor grade was an independent risk factor. CONCLUSION: Superficial parotidectomy is a reliable surgical procedure associated with a high survival rate and low morbidity in treating APG cancers. The tumor grade is the key prognostic factor.

Effect of Taoren Quyu Decoction on human endometrial cells and its anti-endometriosis activity in rats.

OBJECTIVE: To study the effect of Taoren Quyu Decoction (TQD) on endometrial cells in patients with endometriosis (EMs) and EMs in rats. METHODS: A total of 60 female Wistar rats were randomly divided into 4 groups, namely, normal group, model group, positive group and TQD group, each group having 15 rats. Except the normal group, EMs model was established in the other three groups by transplanting the rat autologous endometrium. After 4 weeks of intragastric administration, blood, eutopic and ectopic endometrial tissues of rats in each group were collected to detect the serum levels of estrogen (E2), cancer antigen 125 (CA125), endometrial antibody (EMAb), and expressions of microvessel density (MVD), vascular endothelial growth factor (VEGF) and angiopoietin (Ang-2). The volume of endometriosis cyst was determined simultaneously. For the in vitro culture of human endometrial cells, 4 groups, namely, normal group, model group, positive group and TQD group were used. The positive group and TQD group were treated with danazol and TQD respectively. Then 24 h after the treatment, the expressions of survivin and tumor suppressor gene (p53) of each group were detected. RESULTS: The volumes of the endometriosis cysts in the positive group and the TQD group were significantly reduced compared with the model group (P < 0.05). The serum levels of E2, CA125 and EMAb, and the expressions of MVD, VEGF and Ang-2 in the model group were significantly increased compared with the normal group (P < 0.05); while they were all significantly reduced in the positive group and TQD group (P < 0.05). Compared with the normal group, the expression of survivin in the model group was significantly up-regulated (P < 0.05), and expression of p53 was significantly reduced (P < 0.05); compared with the model group, the expressions of survivin in the positive and TQD groups were significantly decreased (P < 0.05), and expression of p53 was significantly up-regulated (P < 0.05). The difference between positive group and TQD group was not statistically significant (P > 0.05). CONCLUSIONS: TQD has a significant anti-EMs effect, and its mechanism of action may be related to anti-angiogenesis and promoting apoptosis of ectopic endometrial cell.

MicroRNA-216a promotes the metastasis and epithelial-mesenchymal transition of ovarian cancer by suppressing the PTEN/AKT pathway.

MicroRNAs, a group of posttranscriptional regulators of numerous genes, are active participators during the development and progression of ovarian cancer (OC). This study confirmed for the first time that miR-216a was gradually increased in normal, benign, borderline, and OC tissues and that its expression was significantly upregulated in all OC cell lines. Analysis of its clinical association demonstrated that elevated expression of miR-216a was associated with lymph node metastasis and advanced FIGO stage and was correlated with the poor survival of OC patients. Functional experiments showed that miR-216a overexpression potentiated the migration and invasion of CAOV3 cells while miR-216a inhibition reduced the migration and invasion of SKOV-3 cells. Both gain and lose of function assay showed that miR-216a promoted epithelial-mesenchymal transition (EMT) of OC cells. Mechanistically, phosphatase and tensin homolog (PTEN) was confirmed as a direct downstream target of miR-216a in OC cells. Alerting miR-216a expression in OC cells modulated the activity of PTEN/AKT pathway in OC cells. Furthermore, this study confirmed that miR-216a exerted its promoting effects on the metastatic behaviors and EMT of OC cells by inhibiting PTEN/AKT pathway. Taken together, this study demonstrates that miR-216a exerts a promoting role in the metastasis of OC and can serve as a promising biomarker and an attractive therapeutic target of OC.

Streptomyces luozhongensis sp. nov., a novel actinomycete with antifungal activity and antibacterial activity.

A novel actinomycete strain, designated TRM 49605T, was isolated from a desert soil sample from Lop Nur, Xinjiang, north-west China, and characterised using a polyphasic taxonomic approach. The strain exhibited antifungal activity against the following strains: Saccharomyces cerevisiae, Curvularia lunata, Aspergillus flavus, Aspergillus niger, Fusarium oxysporum, Penicillium citrinum, Candida albicans and Candida tropicalis; Antibacterial activity against Bacillus subtilis, Staphylococcus epidermidis and Micrococcus luteus; and no antibacterial activity against Escherichia coli. Phylogenetic analysis based on 16S rRNA gene sequences affiliated strain TRM 49605T to the genus Streptomyces. Strain TRM 49605T shows high sequence similarities to Streptomyces roseolilacinus NBRC 12815T (98.62 %), Streptomyces flavovariabilis NRRL B-16367T (98.45 %) and Streptomyces variegatus NRRL B-16380T (98.45 %). Whole cell hydrolysates of strain TRM 49605T were found to contain LL-diaminopimelic acid as the diagnostic diamino acid and galactose, glucose, xylose and mannose as the major whole cell sugars. The major fatty acids in strain TRM 49605T were identified as iso C16:0, anteiso C15:0, C16:0 and Summed Feature 5 as defined by MIDI. The main menaquinones were identified as MK-9(H4), MK-9(H6), MK-9(H8) and MK-10(H6). The polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine, phosphatidylinositol and phosphatidylinositol mannoside. The G+C content of the genomic DNA was determined to be 71.2 %. The DNA-DNA relatedness between strain TRM 49605T and the phylogenetically related strain S. roseolilacinus NBRC 12815T was 60.12 ± 0.06 %, which is lower than the 70 % threshold value for delineation of genomic prokaryotic species. Based on the phenotypic, chemotaxonomic and phylogenetic data, strain TRM 49605T (=CCTCC AA2015026T = KCTC 39666T) should be designated as the type strain of a novel species of the genus Streptomyces, for which the name Streptomyces luozhongensis sp. nov. is proposed.

Streptomyces canalis sp. nov., an actinomycete isolated from an alkali-removing canal.

A novel actinomycete strain, designated TRM 46794-61T, was isolated from an alkali-removing canal in 14th Farms of Xinjiang Production and Construction Corps, north-west China. The isolate contained ll-diaminopimelic acid as the diagnostic diamino acid. The whole-cell sugar patterns of the isolate contained ribose, mannose and glucose. The polar lipids were diphosphatidylglycerol, phosphatidylmethylethanolamine, phosphatidylethanolamine, phosphatidylcholine, phosphatidylinositol, phosphatidylinositol mannoside and two unidentified phospholipids. The predominant menaquinones were MK-9(H2), MK-9(H4), MK-9(H6) and MK-9(H8). The major fatty acids were iso-C16 : 0, anteiso-C17 : 0 and anteiso-C15 : 0. The G+C content of the DNA was 70.4 mol%. Phylogenetic analysis showed that strain TRM 46794-61T had a 16S rRNA gene sequence similarity of 97.6 % with the most closely related species with a validly published name, Streptomyces aidingensis TRM 46012T, and it could be distinguished from all species in the genus Streptomyces based on data from this polyphasic taxonomic study. However, DNA-DNA hybridization studies between strain TRM 46794-61T and S.aidingensis TRM 46012T showed only 45.4 % relatedness. On the basis of these data, strain TRM 46794-61T should be designated as a representative of a novel species of the genus Streptomyces, for which the name Streptomyces canalis sp. nov. is proposed. The type strain is TRM 46794-61T (=CCTCC AA 2015006T=KCTC 39568T).

Impact of the International Association for the Study of Lung Cancer/American Thoracic Society/European Respiratory Society classification of stage IA adenocarcinoma of the lung: Correlation between computed tomography images and EGFR and KRAS gene mutations.

The aim of the present study was to compare pathological diagnoses, as determined by the new International Association for the Study of Lung Cancer/American Thoracic Society/European Respiratory Society (IASLC/ATS/ERS) classification, with conventional radiological features. In addition, the present study aimed to evaluate the correlation among clinical characteristics, computed tomography (CT) images and gene mutation status in patients with stage IA adenocarcinoma of the lung. A total of 212 patients with stage IA lung adenocarcinoma were included in the study. The patients were classified into pure ground-glass opacity (pGGO), mixed GGO (mGGO) and solid GGO (sGGO) by CT imaging. Histological subtype was classified according to the IASLC/ATS/ERS classification of lung adenocarcinoma. In addition, epidermal growth factor receptor (EGFR) and Kirsten rat sarcoma (KRAS) mutation assays were performed, and 36.8% of patients (78/212) were determined to have an EGFR mutation, while 8.5% of patients (18/212) were found to have a KRAS mutation. According to the IASLC/ATS/ERS classification, 44 cases were diagnosed as adenocarcinoma in situ (AIS; 20.8%), 62 cases were diagnosed as minimally invasive adenocarcinoma (MIA; 29.2%) and 106 cases were classified as invasive adenocarcinoma (IAC; 50.0%). pGGO image patterns were observed in 39.2% of patients (n=83), while mGGO and sGGO patterns were observed in 28.8% (n=61) and 32.0% (n=68) of patients, respectively. From pGGO to sGGO, cases of AIS and MIA were shown to have a decreasing trend, while IAC cases exhibited an increasing trend (P=0.036). Analysis of the correlation between CT image patterns and gene mutations demonstrated that L858R point mutations, exon 19 deletions and KRAS mutations were more common in lesions with a lower GGO proportion (P=0.029, 0.027 and 0.018, respectively). Therefore, according to the IASLC/ATS/ERS classification, GGO imaging patterns were shown to correlate with subtypes of adenocarcinomas. In addition, EGFR and KRAS mutations were found to be associated with lesions with a low GGO proportion. Therefore, analysis of GGO lesions may offer useful indications of the histological subtype of an adenocarcinoma in patients with stage IA lung adenocarcinoma, and predictive value for EGFR and KRAS mutations.

Glycomyces fuscus sp. nov. and Glycomyces albus sp. nov., actinomycetes isolated from a hypersaline habitat.

Two actinomycete strains, designated TRM 49117(T) and TRM 49136(T), were isolated from a hypersaline habitat in Xinjiang Province, north-west China and were characterized taxonomically by using a polyphasic study. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain TRM 49117(T) had 93.93% similarity with the type strain Glycomyces halotolerans TRM 40137(T) (GenBank accession no. HQ651156) and TRM 49136(T) had 94.32% similarity with G. halotolerans TRM 40137(T). The 16S rRNA gene sequence similarity between the two new isolates was 93%. The isolates contained meso-diaminopimelic acid as the diagnostic diamino acid and anteiso-C15 : 0, iso-C16 : 0 and anteiso-C17 : 0 as major cellular fatty acids. The predominant menaquinones of the isolates were MK-9(H4) and MK-9(H6). The whole-cell sugar patterns of these strains contained xylose and ribose, and strain TRM 49136(T) also contained arabinose. The polar lipid pattern of strain TRM 49117(T) comprised phosphatidylglycerol, diphosphatidylglycerol, phosphatidylinositol mannosides, phosphatidylcholine, phosphatidylinositol and three additional unknown phospholipids. The polar lipid pattern of strain TRM 49136(T) comprised phosphatidylglycerol, phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylinositol, glycolipids and two phosphoglycolipids of unknown composition. Genotypic and phenotypic data confirmed that strains TRM 49117(T) and TRM 49136(T) represent two novel species, clearly different from related species of the genus Glycomyces, for which the names Glycomyces fuscus sp. nov. (type strain TRM 49117(T) = CCTCC AA 2013003(T) = NRRL B-59998(T) = KACC 17682(T)) and Glycomyces albus sp. nov. (type strain TRM 49136(T) = CCTCC AA 2013004(T) = NRRL B-24927(T) = KACC 17681(T)) are proposed.