ACSL4 accelerates osteosarcoma progression via modulating TGF-ß/Smad2 signaling pathway.

Osteosarcoma (OS) is a malignant bone sarcoma arising from mesenchymal stem cells. The biological role of Acyl-CoA synthetase long-chain family member 4 (ACSL4), recently identified as an oncogene in numerous tumor types, remains largely unclear in OS. In this study, we investigated the expression of ACSL4 in OS tissues using immunohistochemistry staining (IHC) staining of a human tissue microarray and in OS cells by qPCR assay. Our findings revealed a significant up-regulation of ACSL4 in both OS tissues and cells. To further understand its biological effects, we conducted a series of loss-of-function experiments using ACSL4-depleted MNNG/HOS and U-2OS cell lines, focusing on OS cell proliferation, migration, and apoptosis in vitro. Our results demonstrated that ACSL4 knockdown remarkably suppressed OS cell proliferation, arrested cells in the G2 phase, induced cell apoptosis, and inhibited cell migration. Additionally, a subcutaneous xenograft mice model was established to validate the in vivo impact of ACSL4, revealing ACSL4 silencing impaired tumor growth in the OS xenograft mice. Additionally, we discovered that ACSL4 could regulate the phosphorylation level of Smad2 through cooperative interactions, and treatment with a TGF-ß inhibitor weakened the promoting effects of ACSL4 overexpression. In short, ACSL4 regulated OS progression by modulating TGF-ß/Smad2 signaling pathway. These findings underscore ACSL4 as a promising therapeutic target for OS patients and contribute novel insights into the pathogenesis of OS.

Fe2+-NTA synergized UV254 photolytic defluorination of perfluorooctane sulfonate (PFOS): Enhancing through intramolecular electron density perturbation via electron acquisition.

Perfluorooctane sulfonate (PFOS) is a persistent organic pollutant posing a risk in environmental persistence, bioaccumulation and biotoxicity. This study was to reach a comprehensive and deeper understanding of PFOS elimination in a UV254 photolytic treatment with the co-presence of Fe2+ and nitrilotriacetic acid trisodium salt (NTA). PFOS defluorination was noticeably enhanced in the UV/Fe2+-NTA treatment compared with UV/NTA, UV/Fe2+ and our previously studied UV/Fe3+ treatments. UV-vis, FTIR, and UPLC/MS-MS results indicated the formation of PFOS-Fe2+-NTA complex in PFOS, Fe2+ and NTA mixture. The transition energy gap of PFOS-Fe2+-NTA decreased below the excitation energy supplied by UV254 irradiation, corresponding with red shift appearing in UV-vis scanning spectrum. This favored intramolecular electron transfer from Fe2+-NTA to PFOS under UV254 irradiation to form electron-accepting PFOS. Molecular electrostatic potential and atom charge distribution analyses suggested electron density rearrangement and perturbation in the perfluorinated carbon chain of electron-accepting PFOS, leading to the decrease in bond dissociation energies. Intermediate products detection suggested the parallel defluorination pathways of PFOS desulfonation, middle carbon chain scission and direct C-F cleavage. NTA exhibited crucial functions in the UV/Fe2+-NTA treatment by holding Fe2+/Fe3+ in soluble form as a chelant and favoring water activation to generate hydrated electrons (eaq-) under UV irradiation as a photosensitizer. Fe2+ acting as the conduit for electron transfer and the bridge of PFOS anion and NTA was thought functioning best at 200 µM in this study. The degree of UV/Fe2+-NTA -synergized PFOS defluorination also depended on eaq- yield and UV254 photon flux. The structure dependence on the electron transfer process of PFOS and PFOA was explored incorporating molecular structure descriptors. Because of possessing greater potential to acquire electrons or less likeliness to donate its electrons than PFOA, PFOS exhibited faster defluorination kinetics in the published "reduction treatments" than "oxidation" ones. Whereas, PFOA defluorination kinetics were at similar level in both "reduction" and "oxidation" treatments.

Changes in microcystin concentration in Lake Taihu, 13 years (2007-2020) after the 2007 drinking water crisis.

Since the 2007 water crisis occurred in Lake Taihu, substantial measures have been taken to restore the lake. This study evaluates the effectiveness of these restoration activities. We examined the physicochemical parameters and the distribution of microcystin and Microcystis in both the water column and sediment during the bloom period of May 2020 to October 2020. The mean value of extracellular and intracellular microcystin content was 0.12 µg L-1 and 16.26 µg L-1, respectively. The mean value of microcystin in sediment was 172.02 ng g-1 and peaked in August. The concentration in the water and sediment was significantly lower than the historical average concentration. The abundance of toxigenic Microcystis and total Microcystis in the water column ranged from 2.61 × 102 to 2.25 × 109 copies·L-1 and 8.28 × 105 to 2.76 × 109 copies·L-1, respectively. The proportion of toxic Microcystis in the sediment ranging from 31.2% to 19.12%. The highest and lowest region was Meiliang Bay and Grass-algae type zone, respectively. The copy number of the 16S rRNA gene was 1-4 orders of magnitude higher than that of mcyA gene in populations of Microcystis, indicating that non-toxic Microcystis was the dominant form in the majority of the lake. The abundance of toxic Microcystis in the water column was positively correlated with total phosphorus, PO43--P and pH, while the water temperature played distinct role to the distribution of toxic Microcystis in sediment. Our research indicated phosphorus remains a key factor influencing the toxic Microcystis and microcystins in the water column. pH played distinct roles in the distribution of microcystins in sediment and water column. The increasing water temperature is a threat. Explicit management actions and policies, which take into account nutrient concentrations, pH, and increasing temperatures, are necessary to understand and control the distribution of microcystin and Microcystis in Lake Taihu.

Preliminary evidence for endoscopic surgery combined with postoperative anti-PD-1 immunotherapy in advanced recurrent nasopharyngeal carcinoma.

BACKGROUD: Endoscopic surgery can be used as the main treatment for advanced recurrent nasopharyngeal carcinoma (rNPC). However, there is a huge clinical controversy about the need for consolidated immunotherapy after surgery. METHODS: We performed a retrospective propensity score-matched analysis (1:2) of patients with locally advanced rNPC who underwent endoscopic nasopharyngectomy (ENPG) combined with anti-programmed cell death protein-1 (PD-1) monotherapy or ENPG alone. The survival rate was analyzed by Kaplan-Meier method. The primary endpoint was progression-free survival (PFS). The secondary endpoints included overall survival (OS), objective response rate (ORR) and disease control rate (DCR). Potential surgical-related complications and immune-related adverse events (AEs) were also assessed. RESULTS: We recruited 10 patients receiving ENPG plus anti-PD-1 monotherapy and 20 receiving ENPG alone. During the mean follow-up of 23.8 months, a significant improvement in the 2-year PFS was detected in the consolidation immunotherapy group compared to the ENPG alone group (80.0% vs. 40.0%; HR = 0.258; 95% CI: 0.09-0.72; p = 0.04), while the 2-year OS in the consolidation immunotherapy group was not significantly longer than that in the ENPG alone group (90.0% vs. 75.0%; HR = 0.482; 95% CI: 0.08-3.00; p = 0.50). The incidence of surgical-related complications in the consolidation immunotherapy group and ENPG alone group was 70.0 and 60.0%, respectively. Immune-related AEs were similar between the toripalimab arm (75.0%) and the camrelizumab arm (66.7%). Surgical-related complications depend on symptomatic treatments. Immune-related AEs were mild and tolerable. CONCLUSIONS: Consolidation immunotherapy regimen for patients with advanced rNPC after ENPG compared to ENPG alone provides a superior PFS rate with a manageable safety profile.

miRNA-19a exerts an anti-apoptotic effect in spinal cord injured rats via the PTEN pathway.

Introduction: Contusion spinal cord injury is involved in a number of cellular, biochemical and molecular changes. We studied the overall expression pattern of miRNAs on day 1 and 3 after spinal cord injury and the involved pathways. Material and methods: A spinal cord injury model was developed by contusion injury in rats. Microarray analysis and qRT-PCR were done for expression of miRs. The Basso, Beattie and Bresnahan (BBB) locomotor score was determined after spinal injury. Lesions at the injured site were analyzed by cresyl staining. Western blot analysis was carried out to analyze protein levels. Immunohistochemical staining was done to evaluate immunoreactivity. TUNEL assay was performed to determine the number of apoptotic cells. Results: The microarray analysis data suggested that about eight miRs were upregulated whereas four were downregulated in rats subjected to spinal cord injury on day 1. On comparing sham operated rats from the day 3 group two miRs were overexpressed and four were downregulated. miR-19a was the most deregulated. miR-19a antagomir was used as an inhibitor, which aggravated the functional deficit, decreased the protection of spinal cord tissue and elevated the number of apoptotic cells. The treatment of miR-19a antagomir increased the expression of FasL along with PTEN, but it failed to affect the levels of PDCD4. Conclusions: The results suggested that miR-19a plays a potential role in halting the neuronal cell death spinal cord injury and that the protective role of miR-19a may be due to its regulatory effect on pro-apoptotic genes.

[Research of epigallocatechin gallate in delaying chondrocyte senescence].

Objective: To investigate the effect of epigallocatechin gallate (EGCG) on chondrocyte senescence and its mechanism. Methods: The chondrocytes were isolated from the articular cartilage of 4-week-old Sprague Dawley rats, and cultured with type Ⅱcollagenase and passaged. The cells were identified by toluidine blue staining, alcian blue staining, and immunocytochemical staining for type Ⅱ collagen. The second passage (P2) cells were divided into blank control group, 10 ng/mL IL-1ß group, and 6.25, 12.5, 25.0, 50.0, 100.0, and 200.0 µmol/L EGCG+10 ng/mL IL-1ß group. The chondrocyte activity was measured with cell counting kit 8 after 24 hours of corresponding culture, and the optimal drug concentration of EGCG was selected for the subsequent experiment. The P2 chondrocytes were further divided into blank control group (group A), 10 ng/mL IL-1ß group (group B), EGCG+10 ng/mL IL-1ß group (group C), and EGCG+10 ng/mL IL-1ß+5 mmol/L 3-methyladenine (3-MA) group (group D). After cultured, the degree of cell senescence was detected by ß-galactosidase staining, the autophagy by monodansylcadaverine method, and the expression levels of chondrocyte-related genes [type Ⅱ collagen, matrix metalloproteinase 3 (MMP-3), MMP-13] by real-time fluorescent quantitative PCR, the expression levels of chondrocyte-related proteins (Beclin-1, LC3, MMP-3, MMP-13, type Ⅱ collagen, P16, mTOR, AKT) by Western blot. Results: The cultured cells were identified as chondrocytes. Compared with the blank control group, the cell activity of 10 ng/mL IL-1ß group significantly decreased ( P<0.05). Compared with the 10 ng/mL IL-1ß group, the cell activity of EGCG+10 ng/mL IL-1ß groups increased, and the 50.0, 100.0, and 200.0 µmol/L EGCG significantly promoted the activity of chondrocytes ( P<0.05). The 100.0 µmol/L EGCG was selected for subsequent experiments. Compared with group A, the cells in group B showed senescence changes. Compared with group B, the senescence rate of chondrocytes in group C decreased, autophagy increased, the relative expression of type Ⅱ collagen mRNA increased, and relative expressions of MMP-3 and MMP-13 mRNAs decreased; the relative expressions of Beclin-1, LC3, and type Ⅱ collagen proteins increased, but the relative expressions of P16, MMP-3, MMP-13, mTOR, and AKT proteins decreased; the above differences were significant ( P<0.05). Compared with group C, when 3-MA was added in group D, the senescence rate of chondrocytes increased, autophagy decreased, and the relative expressions of the target proteins and mRNAs showed an opposite trend ( P<0.05). Conclusion: EGCG regulates the autophagy of chondrocytes through the PI3K/AKT/mTOR signaling pathway and exerts anti-senescence effects.

[The outcome of transnasal endoscopic total maxillectomy in the treatment of sinonasal adenoid cystic carcinoma].

Objective:To assess the prognosis of sinonasal adenoid cystic carcinoma with hard palatine invasion treated by transnasal endoscopic total maxillectomy. Methods:Clinical data of twenty-six patients with sinonasal adenoid cystic carcinoma invading hard palatine treated by transnasal endoscopic total maxillectomy between May 2014 and December 2020 was analyzed retrospectively. Survival rate, local recurrence and distant metastasis were analyzed using Kaplan-Meier method. Cox regression was used to investigate the prognosis factors. Masticatory function after maxillectomy has also been assessed using the questionnaire of patients' satisfaction about masticatory function. Results:Margins in 8 patients（30%） were positive. The median time of follow-up was 38 months（6 to 85 months）. Twenty-five patients recurred. Four patients died of distant metastasis. The 5-year overall survival rate and relapse-free survival rate was 79.5% and 89.1%, respectively. Independent predictors of outcome on multivariate analysis were positive margin（P=0.018）, recurrence（P=0.006） and distant metastasis（P=0.04）. Conclusion:Transnasal endoscopic total maxillectomy could be performed for the treatment of the sinonasal adenoid cystic carcinoma with hard palatine invasion. Positive margin, local recurrence and distant metastasis were important predictors for patients' prognosis.

COLEC10 Induces Endoplasmic Reticulum Stress by Occupying GRP78 and Inhibits Hepatocellular Carcinoma.

Collectin subfamily member 10 (COLEC10), a C-type lectin mainly expressed in the liver, is involved in the development of hepatocellular carcinoma (HCC). However, its underlying molecular mechanism in HCC progression remains unknown. In this study, reduced COLEC10 expression in tumor tissues was validated using various HCC cohorts and was associated with poor patient prognosis. COLEC10 overexpression attenuated HCC cell growth and migration abilities in vitro and in vivo. We identified that COLEC10 was a novel interactor of 78-kDa glucose-regulated protein (GRP78), a master modulator of the unfolded protein response in the endoplasmic reticulum (ER). COLEC10 overexpression potentiated ER stress in HCC cells, as demonstrated by elevated expression levels of phosphorylated protein kinase RNA-like ER kinase, phosphorylated inositol-requiring protein 1α, activating transcription factor 4, DNA damage-inducible transcript 3, and X-box-binding protein 1s. The ER in COLEC10-overexpressing cells also showed a dilated and fragmented pattern. Mechanistically, COLEC10 overexpression increases GRP78 occupancy through direct binding by the C-terminal carbohydrate recognition domain in the ER, which released and activated the ER stress transducers protein kinase RNA-like ER kinase and phosphorylated inositol-requiring protein 1α, triggering the unfolded protein response activity. COLEC10-overexpressing HCC cells generated a relatively high reactive oxygen species level and switched to apoptotic cell death under sorafenib-treated conditions. Our study provides the first novel view that COLEC10 inhibits HCC progression by regulating GRP78-mediated ER stress signaling and may serve as a promising therapeutic and prognostic biomarker.

Endoscopic surgery is superior to intensity-modulated radiotherapy in the treatment of advanced recurrent nasopharyngeal carcinoma.

BACKGROUND: The choice between endoscopic surgery and re-radiotherapy as the main treatment modality in patients with advanced recurrent nasopharyngeal carcinoma (rNPC) remains controversial. Therefore, in this study, we compared the efficacies of endoscopic surgery and intensity-modulated radiotherapy (IMRT) in patients with rNPC. METHODS: All patients with advanced rNPC (T3 and T4) who underwent salvage treatment were enrolled from January 2009 to December 2020. Overall survival (OS) was analyzed using a log-rank analysis. Univariate and multivariate analyses of OS were performed using a Cox regression model. Common treatment-related complications of endoscopic surgery were compared with those of IMRT. RESULTS: The numbers of patients with T3 and T4 tumors were 163 (64.2%) and 91 (35.8%), respectively; 192 patients underwent endoscopic surgery, 51 received IMRT, and 11 received three-dimensional conformal radiotherapy (3D-CRT). The 3-year OS of patients treated with endoscopic surgery was 59.3%, which was significantly higher than that of patients treated with IMRT (34.7%, p < 0.001) or 3D-CRT (43.6%, p = 0.012). Multivariate analyses showed that IMRT was an independent risk factor for OS compared with endoscopic surgery (hazard ratio, 2.068; 95% confidence interval, 1.395-3.069, p < 0.001). Complications of aural fullness (p = 0.001), nasopharyngeal necrosis (p = 0.004), nasopharyngeal hemorrhage (p = 0.004), dysphagia (p < 0.001), and cerebral infarction (p = 0.030) were significantly lower in the endoscopic surgery group than in the IMRT group. CONCLUSION: Endoscopic surgery may be a more promising salvage treatment than IMRT to maximize survival and minimize treatment-related complications in advanced rNPC. These results will be significant in deciding the optimal treatment for patients with advanced rNPC.

Efficient degradation and deiodination of iopamidol by UV/sulfite process: Assessment of typical process parameters and transformation paths.

Iopamidol (IPM) is widely used in medical clinical examination and treatment and has immeasurable harm to the ecological environment. The combination of UV and sulfite (UV/sulfite) process was developed to degrade IPM in this study. In contrast to that almost no removal of IPM was observed under sulfite reduction alone, the UV/sulfite process could efficiently reductively degrade IPM with the observed rate constant (kobs) of 2.08 min-1, which was nearly 4 times that of UV irradiation alone. The major active species in the UV/sulfite process were identified as hydrated electrons (eaq-) by employing active species scavengers. The influence of the initial pH, sulfite dosage, IPM concentration, UV intensity and common water matrix were evaluated. The degradation of IPM reached nearly 100% within only 2.5 min at pH 9, and kobs increased at higher initial sulfite dosages and greater UV intensities. HCO3- had a limited effect on the degradation of IPM, while humic acid (HA) was found to be a strong inhibitor in the UV/sulfite process. With the synergistic action of UV/sulfite, most of the iodine in IPM was found to release in the form of iodide ions (up to approximately 98%), and a few formed iodide-containing organic compounds, reducing significantly the toxicity of degradation products. Under direct UV irradiation and free radical reduction (mainly eaq-), 15 transformation intermediates of IPM were produced by amide hydrolysis, deiodination, hydroxyl radical addition and hydrogen abstraction reactions, in which free radical attack accounted for the main part. Consequently, the UV/sulfite process has a strong potential for IPM degradation in aquatic environments.

Transinfratemporal Fossa Transposition of the Temporalis Muscle Flap for Skull Base Reconstruction after Endoscopic Expanded Nasopharyngectomy: Anatomical Study and Clinical Application.

Objective Temporalis muscle flap (TMF) is widely used in traditional skull base surgery, but its application in endoscopic skull base surgery remains rarely reported. We aimed to investigate the surgical anatomy and clinical application of TMF for reconstruction of skull base defects after expanded endoscopic nasopharyngectomy. Methods Nine fresh cadaver heads (18 sides) were used for endoscopic dissection at the University of Pittsburgh School of Medicine in the United States. TMF was harvested using a traditional open approach and then transposed into the maxillary sinus and nasal cavity through the infratemporal fossa using an endoscopic transnasal transmaxillary approach. TMF length was then measured. Moreover, TMF was used for the reconstruction of skull base defects of six patients with recurrent nasopharyngeal carcinoma after expanded endoscopic nasopharyngectomy. Results The length of TMF harvested from the temporal line to the tip of the coronoid process of the mandible was 11.8 ± 0.9 cm. The widest part of the flap was 9.0 ± 0.4 cm. When TMF was dislocated from the coronoid process of the mandible, approximately another 2 cm of reach could be obtained. When the superficial layer of the temporalis muscle was split from the deep layer, the pedicle length could be extended 1.9 ± 0.2 cm. TMF could cover skull base defects in the anterior skull base, sellar, and clivus regions. Conclusion TMF can be used to reconstruct skull base defects after endoscopic expanded nasopharyngectomy and can effectively prevent the occurrence of serious complications in patients with recurrent nasopharyngeal carcinoma.

New insights into ferric iron-facilitated UV254 photolytic defluorination of perfluorooctanoic acid (PFOA): Combined experimental and theoretical study.

UV/Fe3+-facilitated PFOA defluorination was often reported and recognized to proceed through a "ligand-to-metal charge transfer (LMCT)" mechanism in the literatures. Sufficient Fe3+ supply is important for sustaining the LMCT reaction pathway. In this study, an interesting "excessive defluorination" was observed, even the continuous Fe3+ supply was cut off, implying a parallel mechanism strengthening PFOA defluorination. Based the results of intermediate products detection, 19F NMR analysis, and exploration of electron density alternation, transition energy evolution, and bonds characteristics, remarkable electron density perturbation in [PFOA-Fe]2+ was revealed. This effect was triggered by the complexation between PFOA anion and Fe3+, diminishing electron shielding on the perfluorinated carbon chain. Hence, the dissociation energy of C-C bonds was reduced by maximally 53% (C4-C5). Once attacked by high-flux UV254 photons, the perfluorinated carbon chain underwent scission, and subsequent defluorination was achieved via hydrolysis reactions. This parallel mechanism cooperated with the LMCT mechanism, leading to the observed "excessive defluorination." The degree of UV/Fe3+-synergized PFOA defluorination depended on UV254 photon flux and Fe3+ dosage. High UV254 intensity guaranteed fast defluorination kinetics. A [Fe3+]/[PFOA] molar ratio near 1 showed the best UV/Fe3+ synergic effect on PFOA defluorination.

Chloride intercellular channel 3 suppression-mediated macrophage polarization: a potential indicator of poor prognosis of hepatitis B virus-related acute-on-chronic liver failure.

Patients with hepatitis B virus-related acute-on-chronic liver failure (HBV-ACLF) are characterized by immune paralysis and susceptibility to infections. Macrophages are important mediators of immune responses can be subclassified into two main phenotypes: classically activated and alternatively activated. However, few studies have investigated changes to macrophage polarization in HBV-related liver diseases. Therefore, we investigated the functional status of monocyte-derived macrophages (MDMs) from patients with mild chronic hepatitis B (n = 226), HBV-related compensated cirrhosis (n = 36), HBV-related decompensated cirrhosis (n = 40), HBV-ACLF (n = 62) and healthy controls (n = 10), as well as Kupffer cells (KCs) from patients with HBV-ACLF (n = 3). We found that during the progression of HBV-related liver diseases, the percentage of CD163+ CD206+ macrophages increased, while the percentage of CD80+ human leukocyte antigen-DR+ macrophages decreased significantly. MDMs and KCs mainly exhibited high CD163+ CD206+ expression in patients with HBV-ACLF, which predicted poor clinical outcome and higher liver transplantation rate. Transcriptome sequencing analysis revealed that chloride intracellular channel-3 (CLIC3) was reduced in patients with HBV-ACLF, indicating a poor prognosis. To further study the effect of CLIC3 on macrophage polarization, human monocytic THP-1 cell-derived macrophages were used. We found that classical and alternative macrophage activation occurred through nuclear factor kappa B (NF-κB) and phosphoinositide 3-kinase/protein kinase B pathways, respectively. CLIC3 suppression inhibited NF-κB activation and promoted the alternative activation. In conclusion, macrophage polarization gradually changed from classically activated to alternatively activated as HBV-related liver diseases progressed. Both CLIC3 suppression and increased alternatively activated macrophage percentage were potential indicators of the poor prognosis of patients with HBV-ACLF.

Feasibility study on the electrochemical reductive decomposition of PFOA by a Rh/Ni cathode.

The discharge of widely used per- and poly-fluorinated compounds (PFCs) leads to their environmental prevalence, bioaccumulation and biotoxicity; and attracts researches focusing on their treatment in wastewater. Electrochemical reductive treatment is a promising alternative due to its milder reaction conditions and easy operation. The feasibility of electrochemical reductive decomposition of PFOA using a Rh/Ni cathode was explored. The Rh/Ni cathode was fabricated by coating Rh3+ on Ni foil through electrodeposition. The Rh coating was primarily elemental and in a Rh(111) crystalline form. PFOA decomposition and defluorination were observed when using the Rh/Ni cathode where DMF was the solvent and the cathode potential was -1.25 V. A hydrodefluorination reaction was considered having occurred. Because possessing d electrons and empty d orbitals, the Rh coating enhanced PFOA adsorption onto the cathode surface and facilitated CF bond activation through Rh···F interactions. Moreover, the Rh(111) crystal helped chemisorb the generated H* and supply it participating in PFOA decomposition. With the continuous interaction of cathode-supplied electrons, CF bond would ultimately dissociate and transform to CH bond by H* substitution. Adding FeCp2* as a supporting electrolyte enhanced PFOA decomposition by working as the shuttle facilitating PFOA migration to the cathode surface.

Transforming growth factor-ß: An early predictor of a functional cure in chronic hepatitis B treated with interferon.

BACKGROUND: The relationship between the serum transforming growth factor (TGF)-ß level and HBsAg loss has not been clearly elaborated in patients with chronic hepatitis B (CHB). METHODS: Two cohorts of patients with CHB were studied. Cohort A: A total of 207 hepatitis B e antigen (HBeAg)-negative CHB patients who finished ≥1 year nucleos(t)ide analogue monotherapy and sequentially received PEGylated interferon treatment for less than 96 weeks were included. Cohort B: Forty HBeAg-positive patients who initially received entecavir therapy for at least 96 weeks were included. Their viral markers and serum TGF-ß levels were measured at different time points during therapy. RESULTS: The levels of serum TGF-ß and HBsAg (0-24 W) were significantly lower in the patients who had HBsAg< 0.05 IU/mL at 48 weeks than in patients who did not in cohort A. We got the same results when we further divided the patients into subgroups according to the initial HBsAg cut-off values (1000 IU/mL, 100 IU/mL, 50 IU/mL) in cohort A. However, HBeAg seroconversion did not lead to the downregulation of TGF-ß levels. The levels of serum TGF-ß were significantly correlated with HBsAg quantitation in cohort A (12-24 W) but not in cohort B (0-48 W). The levels of TGF-ß at week 12 could be used as an early index to predict a functional cure (AUC=0.818) as well as the levels of HBsAg itself (AUC=0.882) in HBeAg-negative chronic hepatitis B patients treated with PEGylated interferon. CONCLUSIONS: The levels of serum TGF-ß were significantly associated with HBsAg loss but not with HBeAg seroconversion and could be used as an early index to predict a functional cure in CHB patients treated with PEGylated interferon.

Dummy Template-Based Molecularly Imprinted Membrane Coating for Rapid Analysis of Malachite Green and Its Metabolic Intermediates in Shrimp and Fish.

A novel malachite green molecularly imprinted membrane (MG-MIM) with specific selectivity for malachite green (MG) and leucomalachite green (LMG) was prepared using a hydrophobic glass fiber membrane as the polymer substrate, methyl violet as a template analog, 4-vinyl benzoic acid as the functional monomer, and ethyleneglycol dimethacrylate as the crosslinking agent. MG-MIM and non-imprinted membrane (NIM) were structurally characterized using scanning electron microscopy, surface area analyzer, Fourier-transform infrared spectrometer and synchronous thermal analyzer. The results showed that MG-MIM possessed a fluffier surface, porous and looser structure, and had good thermal stability. Adsorption properties of MG-MIM were investigated under optimal conditions, and adsorption equilibrium was reached in 20 min. The saturated adsorption capacities for MG and LMG were 24.25 ng·cm-2 and 13.40 ng·cm-2, and the maximum imprinting factors were 2.41 and 3.20, respectively. Issues such as "template leakage" and "embedding" were resolved. The specific recognition ability for the targets was good and the adsorption capacity was stable even after five cycles. The proposed method was successfully applied for the detection of MG and LMG in real samples, and it showed good linear correlation in the range of 0 to 10.0 µg·L-1 (R2 = 0.9991 and 0.9982), and high detection sensitivity (detection limits of MG and LMG of 0.005 µg/kg and 0.02 µg·kg-1 in shrimp, and 0.005 µg/kg and 0.02 µg/kg in fish sample). The recoveries and relative standard deviations were in the range of 76.31-93.26% and 0.73-3.72%, respectively. The proposed method provides a simple, efficient and promising alternative for monitoring MG and LMG in aquatic products.

Clinical Outcomes of Salvage Endoscopic Nasopharyngectomy for Patients With Advanced Recurrent Nasopharyngeal Carcinoma.

BACKGROUND: Salvage endoscopic nasopharyngectomy has better survival prognosis and fewer complications in the management of early stage rNPC, compared to re-irradiation. However, the treatment modality of advanced recurrent nasopharyngeal carcinoma (rNPC) remains controversial. Thus, the purpose of this study was to investigate the demographics, clinical outcomes, and prognostic factors associated with salvage endoscopic nasopharyngectomy in advanced rNPC. METHODS: This study conducted a retrospective analysis of advanced rNPC patients who underwent salvage surgery betweenm January 2014 and December 2019. The overall survival (OS) and progression-free survival (PFS) were analyzed. Univariable and multivariable analyses of OS and PFS were performed using the Cox regression model. The predicted values of the parameters were determined by means of the receiver operating characteristic (ROC) curve analysis. RESULTS: Among the 120 patients included, there were 75 patients with rT3 stage and 45 patients with rT4 stage. With the median follow-up time of 18 months,the 3 -year OS and PFS were 55.2% and 29.4%, respectively. Multivariate analyses showed that the rNPC patients with older age, low BMI (Body Mass Index), rT4 stage, tumor necrosis, and tumor invasion into the ICA was predictive of worse OS, whereas low BMI and rT4 stage were associated with worse PFS. In addition, the rT stage was identified as a better predictor of OS (area under the ROC curve: 0.669; P=0.003) than the other clinical features. CONCLUSIONS: Salvage treatment using endoscopic nasopharyngectomy appears to be an effective treatment in the management of patients with advanced rNPC. In addition, case matching studies and prospective studies with larger clinical samples are required to further evaluate the efficacy of endoscopic surgery compared with re-irradiation in advanced rNPC.

Expression and clinical significance of inhibitory receptor Leukocyte-associated immunoglobulin-like receptor-1 on peripheral blood T cells of chronic hepatitis B patients: A cross-sectional study.

ABSTRACT: Leukocyte-associated immunoglobulin-like receptor-1 (LAIR-1) is an inhibitory receptor that is expressed on the surface of multiple immune cells and plays key roles in immune modulation. In patients with chronic hepatitis B (CHB), T cell number and functions are abnormal and the expression of inhibitory receptors is elevated. However, the expression of LAIR-1 on T cells in patients with CHB is still undetermined.We recruited 320 patients with CHB in different disease phases and 17 healthy donors. Serum biochemical and virological examinations were performed for each participant, and their demographic and clinical data were collected. According to the latest American Association for the Study of Liver Disease guidelines, we categorized the patients into 4 groups: immune active, immune tolerant, inactive CHB, and gray zone. Additionally, we tested the expression of LAIR-1 on T cells and T cell subsets using flow cytometry.We observed a significant decrease in LAIR-1 expression on CD3+ T cells and its two subsets (CD4+ and CD8+ T cells) in patients with CHB. LAIR-1 expression on T cells was the lowest in the immune active group. LAIR-1 expression levels on CD4+ and CD8+ T cells showed a significant negative association with hepatitis B virus (HBV) DNA load and were lower in hepatitis B e antigen (HBeAg)-positive patients than in HBeAg-negative patients (P < .05). In addition, LAIR-1 expression levels on CD3+, CD4+, and CD8+ T cells were all negatively associated with liver inflammation and fibrosis parameters, such as alanine aminotransferase and aspartate aminotransferase levels, FibroScan value, and aspartate aminotransferase-to-platelet ratio index score.LAIR-1 expression levels on T cells were associated with HBV DNA load and liver inflammation and fibrosis parameters, indicating that LAIR-1 may play an important regulatory role in HBV-induced T cell immune pathogenesis and may be a therapeutic target for CHB.

Serum HBV pregenomic RNA exhibited opposite associations with NKdim and NKbright cell immunity in treatment-naïve chronic hepatitis B patients.

Hepatitis B virus (HBV) pregenomic RNA (pgRNA) is a new biomarker that reflects HBV replication, but its relationship with natural killer (NK) cell immunity in chronic hepatitis B (CHB) is unknown. We assessed serum HBV pgRNA levels in 323 CHB patients by reverse transcription-polymerase chain reaction, assessed cytokine production and activation and inhibitory markers of NK cells by flow cytometry, and measured serum cytokines by enzyme-linked immunosorbent assays (ELISAs). Among the different CHB phases, the serum HBV pgRNA level was highest in the immune-tolerant (IT) and immune-active (IA) phases. Regarding NK and NKdim cells, HBV pgRNA was negatively associated with frequencies, but positively associated with NKp44 and NKp46 expression (activation markers). Regarding NKbright cells, serum HBV pgRNA was positively associated with frequency and programmed cell death protein 1 (PD1) expression (inhibitory marker), but negatively associated with NKp44 and NKp46. Serum HBV pgRNA was not associated with NKp30 (activation marker) on NK cells or subsets. Lastly, serum HBV pgRNA was positively correlated with the levels of serum IL-7 and IL-12P40 (NK cell-promoting cytokines) and negatively correlated with serum prostaglandin E2 (PGE2) level (which negatively regulates NK cells). In conclusion, we found varied relationships between serum HBV pgRNA and NK cells and subsets, indicating that HBV pgRNA may play a complicated role in NK cell-related immunity, providing new information on HBV and host immunity.

Acute and chronic toxicity of microcystin-LR and phenanthrene alone or in combination to the cladoceran (Daphnia magna).

Hazardous substances, such as microcystin-LR (MC-LR) and phenanthrene (Phe) are ubiquitous co-contaminants in eutrophic freshwaters, which cause harms to aquatic organisms. However, the risks associated with the co-exposure of aquatic biota to these two chemicals in the environment have received little attention. In this study, the single and mixture toxic effects of MC-LR and Phe mixtures were investigated in Daphnia magna after acute and chronic exposure. Acute tests showed that the median effective concentrations (48 h) for MC-LR, Phe and their mixtures were 13.46, 0.57 and 8.84 mg/L, respectively. Mixture toxicity prediction results indicated that the independent action model was more applicable than the concentration addition model. Moreover, combination index method suggested that the mixture toxicity was concentration dependent. Synergism was elicited at low concentrations of MC-LR and Phe exposure (≤4.04 + 0.17 mg/L), whereas antagonistic or additive effects were induced at higher concentrations. The involved mechanism of antagonism was presumably attributable to the protective effects of detoxification genes activated by high concentrations of MC-LR in mixtures. Additionally, chronic results also showed that exposure to a MC-LR and Phe mixture at low concentrations (≤50 +2 µg/L) resulted in greater toxic effects on D. magna life history than either chemical acting alone. The significant inhibition on detoxification genes and increased accumulation of MC-LR could be accounted for their synergistic toxic effects on D. magna. Our findings revealed the exacerbated ecological hazard of MC-LR and Phe at environmental concentrations (≤50 +2 µg/L), and provided new insights to the potential toxic mechanisms of MC-LR and Phe in aquatic animals.