Assessment of the efficacy of alkaline water in conjunction with conventional medication for the treatment of chronic gouty arthritis: A randomized controlled study.

BACKGROUND: Chronic gouty arthritis, a prevalent metabolic disorder, has prompted interest in the role of diet and lifestyle in its management. This study examines alkaline water as a non-pharmacological adjunct to traditional medicine, hypothesizing its positive effects on uric acid levels and gout symptoms. METHODS: In this research, 400 chronic arthritis patients from Guangdong Hydropower Hospital (September 2021-September 2023) were randomly assigned to groups receiving varying concentrations of alkaline water alongside conventional Western medicine, or Western medicine alone. A 1-year follow-up involved assessments using visual analogue scales, joint swelling scores, functional assessment scales, and biochemical markers (serum uric acid, creatinine, urea nitrogen) for comprehensive evaluation. RESULTS: Pain relief: High-concentration alkaline water significantly reduced VAS pain scores posttreatment (P < .05). Joint swelling: Greatest improvement observed in high-concentration group (P < .001). Daily activity capability: Notable enhancements in daily activity scores in experimental groups (P < .05). Range of joint motion: All groups showed significant improvement posttreatment (P < .05). Inflammatory markers: Experimental groups experienced a notable decrease in C-reactive protein, especially in the low concentration group (P < .001). Erythrocyte sedimentation rate decreases were marginal and not statistically significant (P > .05). Interleukin-1ß and tumor necrosis factor-α levels significantly decreased, particularly in the low concentration group. Serum uric acid levels: Significant reduction in serum uric acid observed in all alkaline water groups (P < .05), contrasting with the control group. CONCLUSION: Alkaline water, particularly at high concentrations, effectively alleviated pain, reduced joint swelling, enhanced daily activities, and improved joint motion in chronic gouty arthritis treatment. It significantly reduced key inflammatory markers (C-reactive protein, interleukin-1ß, tumor necrosis factor-α) and serum uric acid levels, suggesting its potential as a valuable adjunct in gout management. The limited impact on erythrocyte sedimentation rate warrants further investigation.

Effect of sludge redistribution strategy on stability of partial nitrification-anammox process: Further exploration of the potential value of sludge.

The stability of the two-stage partial nitrification-anammox (PN/A) system was compromised by the inappropriate conversion of insoluble organic matter. In response, a sludge redistribution strategy was implemented. Through the redistribution of PN sludge and anammox sludge in the two-stage PN/A system, a transition was made to the Anammox-single stage PN/A (A-PN/A) system. This specific functional reorganization, facilitated by the rapid reorganization of microbial communities, has the potential to significantly decrease the current risk of suppression. The results of the study showed that implementing the sludge redistribution strategy led to a substantial enhancement in the total nitrogen removal rate (TNRR) by 87.51%, accompanied by a significant improvement of 34.78% in the chemical oxygen demand removal rate (CRR). Additionally, this approach resulted in a remarkable two-thirds reduction in the aeration requirements. High-throughput sequencing revealed that the strategy enriched anammox and ammonia-oxidizing bacteria while limiting denitrifying bacteria, as confirmed by quantitative polymerase chain reaction analysis. Furthermore, the principal component analysis revealed that the location and duration of aeration had direct and indirect effects on functional gene expression and the evolution of microbial communities. This study emphasizes the potential benefits of restructuring microbial communities through a sludge redistribution strategy, especially in integrated systems that encounter challenges with suppression.

How to process synovial fluid samples of gouty arthritis and extract its exosomes for subsequent cytokine analysis.

BACKGROUND: The comparative analysis of ultracentrifugation (UC) and polyethylene glycol (PEG)-based precipitation for the isolation of exosomes in gouty arthritis synovial fluid (GASF) is rarely reported, and it is not known whether different isolation methods can influence subsequent cytokine analysis. METHODS: GA patients were enrolled during a 1-year period from May 2021 to May 2022. Morphology, particle number, size, purity, protein concentration, and biomarker proteins of GASF-derived exosomes in both extraction methods were observed using transmission electron microscopy, nanoparticle tracer analysis, bicinchoninic acid assay, and Western blotting. An ELISA-based assay platform was used to detect the cytokines in exosomes using Meso Scale Discovery. RESULTS: Thirty-two cases of fresh GASF were taken and randomly divided between the UC group (n = 16) and the PEG group (n = 16). Transmission electron microscopy images and nanoparticle tracer analysis results showed round vesicles measuring 100 nm on average. The protein expressions of TSG101, CD63, and CD81 in exosomes of the 2 groups were measured via Western blotting. The number and protein concentration of GASF-derived exosome particles from the PEG group were significantly higher than that of the UC group (P < .001). However, in the purity estimation, the UC group reflected significantly higher exosomes extractability (P < .01). Expression of IL-6 and IL-8 in the GASF-derived exosomes were higher in the UC group (P < .05), showing a median of 3.31 (interquartile range, IQR: 0.84-13.16) pg/mL, and a median of 2.87 (IQR: 0.56-13.17) pg/mL, respectively; moreover, IL-1ß was mostly undetectable in the PEG group. CONCLUSION: The UC method was found to yield exosomes of a higher purity, albeit at a lower quantity but with more abundant inflammatory cytokines; whereas the opposite was the case for the PEG group. The chemical precipitation method might not be suitable in terms of extracting GASF-derived exosomes for inflammation and immunity studies.

LncRNA IUR Is Downregulated in Gastric Carcinoma and Associated with Poor Survival.

Objective: LncRNA IUR has been recently identified as a key regulator of Bcr-Abl-induced tumorigenesis, while its role in gastric carcinoma (GC) is unknown. This study investigated the involvement of IUR in GC. Materials and Methods: Gene expression levels were measured by performing quantitative polymerase chain reaction. Interactions between IUR and ROCK1 were analyzed by transfection experiments. Cell invasion and migration were analyzed by Transwell assay. Results: In this study, the authors showed that IUR was downregulated in GC. A follow-up study showed that low IUR expression levels predicted poor survival. In GC tissues, ROCK1 was upregulated in GC tissues and inversely correlated with IUR. In GC cells, IUR overexpression mediated the downregulation of ROCK1. ROCK1 overexpression resulted in increased GC cell invasion and migration, while IUR overexpression played an opposite role. Conclusion: IUR is downregulated in GC and inhibits GC cell invasion and migration by downregulating ROCK1.

Acupoint application therapy for diarrhea-predominant irritable bowel syndrome: a protocol for systematic review and network meta-analysis.

BACKGROUND: Irritable bowel syndrome is a chronic functional gastrointestinal disease, of which diarrhea-predominant irritable bowel syndrome (IBS-D) is a common subtype. In China, acupoint application therapy is currently widely used as an effective complementary therapy for IBS-D. In the clinical management of IBS-D, acupoint application is usually combined with other therapies, including acupuncture, moxibustion, and Chinese herbal and Western medicine. However, at present, evidence regarding the most effective options for treating IBS-D is insufficient. Therefore, this protocol proposes a systematic review and network meta-analysis for evaluating the effectiveness of acupoint application and its combination therapies in treating IBS-D, and for identifying the acupoint application-related treatments with the highest probability of being the best intervention. METHODS: Six English electronic databases (PubMed, Ovid MEDLINE, Scopus, Web of Science, the Cochrane Library, and EMBASE), four Chinese electronic databases [China National Knowledge Infrastructure (CNKI), China Science and Technology Journal Database (CQVIP), WanFang, and SinoMed), and one Japanese medical database (Citation Information by National Institute of Informatics (CiNii)] will be searched for eligible randomized controlled trials from their inception to June 1, 2022. The efficacy and safety of acupoint application therapy and its combination therapies for patients with IBS-D will be evaluated. The STATA 14.0 (StataCorp, USA) software package will be used for the meta-analysis. A Bayesian network meta-analysis (NMA) will be performed using R (version 4.0.2) and Aggregate Data Drug Information System (ADDIS, version 1.16.8) software packages. Bias risk will be assessed using the Cochrane Collaboration's risk of bias tool; specifically, publication bias will be evaluated using Egger's test and funnel plots. The rank probabilities of various outcomes for each intervention will be calculated, clustered, and ranked using the cumulative ranking curve method. The Grading of Recommendations Assessment, Development, and Evaluation (GRADE) method will be employed to assess the certainty of evidence for NMA outcomes. CONCLUSIONS: This study will aim to determine the clinical efficacy of acupoint application therapy and its combined therapy in the treatment of IBS-D and provide an evidence-based foundation for identifying the best acupoint application program.

Natural lighting enhancing the algae proliferation and nitrogen removal in membrane-aerated bacterial-algal biofilm reactor.

Membrane-aerated bacterial-algal biofilm reactor (MABAR) is an emerging and novel technology in recent years, which has been attracting increasing attention due to its cost-effectiveness and superior removal performance of pollutants by versatile removal pathways in symbiotic bacterial-algal biofilm. However, the wider application of MABAR is hindered by the dilemma of insufficient algae biomass. In this study, an MABAR under natural sunlight was developed and operated for 160 d to access the feasibility of enhancing algae proliferation by natural lighting. Results showed that the MABAR with natural sunlight (nMABAR) demonstrated better performance of pollutants removal. High removal efficiencies of organic matter and NH4-N in nMABAR were 90 % and 92 %, respectively. In particular, the removal efficiency of TN in nMABAR, under less aeration, was up to 80 %, which was 15 % higher than the control reactor. The Chlorophyll-a content indicated that natural sunlight facilitated to algae growth in MABAR, and algae assimilation might be the dominant contributor to NH4-N removal. Moreover, there were microbial shifts in bacterial-algal biofilm in a response to the natural lighting, the nMABAR uniquely possessed a bacterial phylotype termed Thiocapsa, which could play an important role in bacterial nitrification. Algal phylotype Chlorophyceae significantly contributed to pollutants removal and synergistic relationship with bacteria. In addition, the superb performance of nMABAR under less aeration condition suggested that abundant algae were capable of supplying enough O2 for the system. These results provided insight into the natural lighting on algae-bacteria synergistic growth and cost-effective operation strategy for MABAR.

Hot-pressed membrane assemblies enhancing the biofilm formation and nitrogen removal in a membrane-aerated biofilm reactor.

Membrane-aerated biofilm reactor (MABR) is gaining popularity in wastewater treatment as a result of the low-energy delivery of oxygen from the carrier side and reduced sludge waste production, although its wider application suffers from the difficulty in microbial colonization on the smooth, hydrophobic membrane surface. In this study, a newly designed membrane/non-woven fabric assembly, prepared via a facile hot-pressing method, is demonstrated to be efficient in promoting the biofilm formation and nitrogen removal in MABR. The assembly achieved rough surface structure to retain biomass whilst sustained the surface hydrophobicity for a high oxygen transfer ability, which is crucial to support a resilient biofilm. Compared with the slower biomass growth and severe detachment of biofilm in the control, a thicker biofilm was quickly developed on the hot-pressed membrane assembly. High loading rates of organic matter, ammonia nitrogen and total nitrogen (TN) in the MABR using the hot-pressed membrane were 154.9 ± 5.4 g COD/(m2·d), 25.5 ± 0.6 g N/(m2·d) and 22.6 ± 0.7 g N/(m2·d), respectively. Particularly, the removal efficiency of TN was up to 82.8%, which was 2.5 times higher than the control. Furthermore, the biofilm grown on the hot-pressed membrane assembly organized a stable microbial community structure with a steady evolution to achieve a synergistic denitrifying function. Among the bacterial phylotypes, OLB8 might be crucial in denitrification. This study highlighted the significance of this facile membrane modification method to improve the process performance of MABR in wastewater treatment.

Is Chinese herbal formula (nourishing Yin therapy) effective and well tolerated as an adjunct medication to hydroxychloroquine in the treatment of primary Sjögren's syndrome? A meta-analysis of randomised controlled trials.

BACKGROUND: Previously, data mining methodology was used to identify 71 patented prescriptions in Chinese patent databases, indicating that Yin-nourishing therapy (YNT) may be an adjunct medication to hydroxychloroquine in the treatment of primary Sjögren's syndrome (pSS). The purpose of this study was to investigate effects of the addition of YNT, which includes tonifying liver and kidney therapy (TLKT) and replenishing Qi and nourishing Yin therapy (RQNYT), in the treatment of pSS. METHODS: Fourteen databases (including Chinese, English, Japanese, Korean and Latin databases) were searched to identify randomised controlled trials (RCTs) of YNT plus hydroxychloroquine (YNTPH) versus hydroxychloroquine alone in patients with pSS. Relevant publications up to June 2021 were selected. A meta-analysis and trial sequential analysis (TSA) were performed using Review Manager 5.3, Stata 14.0 and TSA 0.9 software. The quality of identified studies was assessed based on the Cochrane risk of bias tool and GRADE (grading of recommendations, assessment, development and evaluation) criteria. RESULTS: We included five RCTs with a total of 345 participants. Pooled results indicated significant differences in short-term outcomes, which were assessed via European League Against Rheumatism (EULAR) Sjögren's Syndrome Patient Reported Index (ESSPRI), EULAR Sjögren's Syndrome Disease Activity Index (ESSDAI), tear production, erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), and immunoglobulin G (IgG) results when YNTPH was compared with hydroxychloroquine use alone (p < 0.05). No significant difference in salivary flow rate was reported. The most common side effect reported for both groups was gastrointestinal reaction. Sensitivity analyses suggested that heterogeneity might be ascribed to clinical methodology. Subgroup analyses revealed heterogeneities regarding salivary flow rate were eliminated. TLKT appeared to be better than RQNYT for improving salivary flow rate. TSA only supported changes in ESSPRI, ESSDAI and ESR values. For all studies, the quality of evidence was low. CONCLUSION: YNTPH may be an effective complementary therapy. Current evidence, however, is insufficient for determining whether YNTPH is more effective than hydroxychloroquine alone. Well-designed RCTs are needed to determine the role of YNT in pSS treatment.

A room temperature all-optical sensor based on two-dimensional SnS2 for highly sensitive and reversible NO2 sensing.

Fiber-optic gas sensors have been considered a low-cost, effective, and robust approach for monitoring nitrogen dioxide (NO2) gas which is a major toxic gaseous pollutant. The integration of functional nanoscale materials provides additional dimensions for realizing ultra-sensitive and selective NO2 detection, however, the trade-off is the need for sophisticated photonic structures or external non-optical peripherals (e.g. electrical heaters). In this work, we demonstrate the development of a room temperature, all-optical, and high-performance NO2 sensor based on a simple D-shaped optical fiber incorporated with ultra-thin two-dimensional (2D) tin disulfide (SnS2). A visible light source at 473 nm is used to power the optical fiber, and at the same time excite the 2D SnS2 layer via the evanescent field, to generate extra charge carriers. Upon exposure to NO2 at room temperature, the physisorbed gas molecules induce charge exchange with the 2D SnS2. This significantly re-distributes the photo-excited charge carriers in the ultra-thin material, therefore manipulating the corresponding optical absorption and scattering. As a result, the optical output power intensity varies as the sensor output through the evanescent field coupling. This all-optical sensor demonstrates an optical power variation of up to 7 µW upon the exposure of NO2 gas at a low concentration of 50 ppb. This response is fully reversible with an extremely low limit of detection (LOD) of 0.464 ppb. We consider that this work provides a feasible and simple solution to realize high-performance optical gas sensors without the integration of external non-optical peripherals for effective monitoring of environmentally hazardous gases.

Hetero-metallic metal-organic frameworks for room-temperature NO2 sensing.

Metal-organic frameworks (MOFs) with exceptional features such as high structural diversity and surface area as well as controlled pore size has been considered a promising candidate for developing room temperature highly-sensitive gas sensors. In comparison, the hetero-metallic MOFs with redox-active open-metal sites and mixed metal nodes may create peculiar surface properties and synergetic effects for enhanced gas sensing performances. In this work, the Fe atoms in the Fe3 (Porous coordination network) PCN-250 MOFs are partially replaced by transition metal Co, Mn, and Zn through a facile hydrothermal approach, leading to the formation of hetero-metallic MOFs (Fe2IIIMII, M = Co, Mn, and Zn). While the PCN-250 framework is maintained, the morphological and electronic band structural properties are manipulated upon the partial metal replacement of Fe. More importantly, the room temperature NO2 sensing performances are significantly varied, in which Fe2Mn PCN-250 demonstrates the largest response magnitude for ppb-level NO2 gas compared to those of pure Fe3 PCN-250 and other hetero-metallic MOF structures mainly attributed to the highest binding energy of NO2 gas. This work demonstrates the strong potential of hetero-metallic MOFs with carefully engineered substituted metal clusters for power-saving and high-performance gas sensing applications.

Heterologous production of α-Carotene in Corynebacterium glutamicum using a multi-copy chromosomal integration method.

The carotenoid, α-carotene, is very beneficial for human health and wellness, but microbial production of this compound is notoriously difficult, due to the asymmetric rings on either end of its terpenoid backbone. Here, we report for the first time the efficient production of α-carotene in the industrial bacterium Corynebaterium glutamicum by using a combined pathway engineering approach including evaluation of the performance of different cyclases and analysis of key metabolic intermediates to determine flux bottlenecks in the carotenoid biosynthesis pathway. A multi-copy chromosomal integration method was pivotal in achieving stable expression of the cyclases. In fed-batch fermentation, 1,054 mg/L of α-carotene was produced by the best strain, which is the highest reported titer achieved in microbial fermentation. The success of increased α-carotene production suggests that the multi-copy chromosomal integration method can be a useful metabolic engineering tool for overexpression of key enzymes in C. glutamicum and other bacterium as well.

Clinical outcomes and patterns of failure of head and neck mucosal melanoma treated with multiple treatment modalities.

OBJECTIVES: The study aims to analyze the clinical characteristics of head and neck mucosal melanoma (MMHN) and the effects of multiple treatment modalities on distant metastasis, recurrence and survival rates to provide a reference for the individualized treatment of MMHN. METHODS: We retrospectively reviewed 262 patients with stage III-IVb MMHN treated from March 1986 to November 2018 at our cancer center. RESULTS: The median follow-up time was 34.0 months (range 1-262 months). The 5-year overall survival (OS), distant metastasis-free survival (DMFS) and disease-free survival (DFS) probabilities were 37.7%, 30.2%, and 20.3%, respectively. The 5-year OS rates for patients with stage III, stage IVA, and stage IVB MMHN were 67.0%, 24.1% and 8.3%, respectively (P < 0.001). A total of 246 (93.9%) patients received surgery, 149 (56.9%) patients received chemotherapy, and 69 (26.3%) patients received immunologic/targeted therapy. A total of 106 (40.5%) patients were treated with radiotherapy: 9 were treated with preoperative radiotherapy, 93 were treated with postoperative radiotherapy, and 4 were treated with radiotherapy alone. In the multivariate Cox regression analysis, primary tumor site, T stage, and immunologic/targeted therapy were independent factors for OS (all P < 0.05). Irradiation technique, T stage, and N stage were independent prognostic factors for DMFS (all P < 0.05). T stage, N stage, and surgery were independent prognostic factors for DFS (all P < 0.05). Distant metastasis was observed in 107 of 262 patients (40.8%), followed by local [74 (28.2%)] and regional [52 (19.8%)] recurrence. CONCLUSIONS: The main reason for treatment failure in MMHN is distant metastasis. Immunologic/targeted therapy and surgery are recommended to improve the survival of MMHN. The American Joint Committee on Cancer (AJCC) 8th edition staging system for MMHN does stage this disease effectively.

Model-based control for column-based continuous viral inactivation of biopharmaceuticals.

Batch low-pH hold is a common processing step to inactivate enveloped viruses for biologics derived from mammalian sources. Increased interest in the transition of biopharmaceutical manufacturing from batch to continuous operation resulted in numerous attempts to adapt batch low-pH hold to continuous processing. However, control challenges with operating this system have not been directly addressed. This article describes a low-cost, column-based continuous viral inactivation system constructed with off-the-shelf components. Model-based, reaction-invariant pH controller is implemented to account for the nonlinearities with Bayesian estimation addressing variations in the operation. The residence time distribution is modeled as a plug flow reactor with axial dispersion in series with a continuously stirred tank reactor, and is periodically estimated during operation through inverse tracer experiments. The estimated residence time distribution quantifies the minimum residence time, which is used to adjust feed flow rates. Controller validation experiments demonstrate that pH and minimum residence time setpoint tracking and disturbance rejection are achieved with fast and accurate response and no instability. Viral inactivation testing demonstrates tight control of logarithmic reduction values over extended operation. This study provides tools for the design and operation of continuous viral inactivation systems in service of increasing productivity, improving product quality, and enhancing patient safety.

Reversible Room Temperature H2 Gas Sensing Based on Self-Assembled Cobalt Oxysulfide.

Reversible H2 gas sensing at room temperature has been highly desirable given the booming of the Internet of Things (IoT), zero-emission vehicles, and fuel cell technologies. Conventional metal oxide-based semiconducting gas sensors have been considered as suitable candidates given their low-cost, high sensitivity, and long stability. However, the dominant sensing mechanism is based on the chemisorption of gas molecules which requires elevated temperatures to activate the catalytic reaction of target gas molecules with chemisorbed O, leaving the drawbacks of high-power consumption and poor selectivity. In this work, we introduce an alternative candidate of cobalt oxysulfide derived from the calcination of self-assembled cobalt sulfide micro-cages. It is found that the majority of S atoms are replaced by O in cobalt oxysulfide, transforming the crystal structure to tetragonal coordination and slightly expanding the optical bandgap energy. The H2 gas sensing performances of cobalt oxysulfide are fully reversible at room temperature, demonstrating peculiar p-type gas responses with a magnitude of 15% for 1% H2 and a high degree of selectivity over CH4, NO2, and CO2. Such excellent performances are possibly ascribed to the physisorption dominating the gas-matter interaction. This work demonstrates the great potentials of transition metal oxysulfide compounds for room-temperature fully reversible gas sensing.

Record-high capture of volatile benzene and toluene enabled by activator implant-optimized banana peel-derived engineering carbonaceous adsorbents.

This work developed a super-high performance of engineering carbonaceous adsorbents from waste banana-peel via an optimized KOH-impregnated approach, which affords outstanding structural property (SBET = 3746.5 m2 g-1, Vtotal = 2.50 cm3 g-1), far outperforming KOH-grinding method-induced counterpart and other known banana peel-derived those. Thereby, this triggers a record-high capture value of volatile organic compounds (VOCs) specific to benzene (27.55 mmol g-1) and toluene (23.82 mmol g-1) in the all known results. The structural expression characters were accurately correlated with excellent adsorption efficiency of VOCs by investigating the synthetic factor-controlling comparative samples. Ulteriorly, the adsorption selectivity prediction at different relative humidity was demonstrated through the DIH (difference of the isosteric heats), highlighting the good superiority in selective adsorption of toluene compared to benzene even under humid atmosphere. Our findings provide the possibility for the practical application and fabrication of waste biomass (banana peel)-derived functional biochar adsorbent in the environmental treatment of threatening VOCs pollutants.

Chimeric Antigen Receptor-T Cells with 4-1BB Co-Stimulatory Domain Present a Superior Treatment Outcome than Those with CD28 Domain Based on Bioinformatics.

BACKGROUND: The second-generation CD19-chimeric antigen receptor (CAR)-T co-stimulatory domain that is commonly used in clinical practice is CD28 or 4-1BB. Previous studies have shown that the persistence of CAR-T in the 4-1BB co-stimulatory domain appears to be longer. METHODS: The expression profile data of GSE65856 were obtained from GEO database. After data preprocessing, the differentially expressed genes (DEGs) between the mock CAR versus CD19-28z CAR T cells and mock CAR versus CD19-BBz CAR T cells were identified using the limma package. Subsequently, functional enrichment analysis of DEGs was performed using the DAVID tool. Then, the protein-protein international (PPI) network of these DEGs was visualized by Cytoscape, and the miRNA-target gene-disease regulatory networks were predicted using Webgestal. RESULTS: A total of 18 common DEGs, 6 CD19-28z specific DEGs and 206 CD19-BBz specific DEGs were identified. Among CD19-28z specific DEGs, down-regulated PAX5 might be an important node in the PPI network and could be targeted by miR-496. In CD19-BBz group, JUN was a hub node in the PPI network and involved in the regulations of miR520D - early growth response gene 3 (EGR3)-JUN and mi-R489-AT-rich interaction domain 5A (ARID5A)-JUN networks. CONCLUSION: The 4-1BB co-stimulatory domain might play in important role in the treatment of CAR-T via miR-520D-EGR3-JUN and miR489-ARID5A-JUN regulation network, while CD28 had a negative effect on CAR-T treatment.

Yeast-Based Synthetic Biology Platform for Antimicrobial Peptide Production.

Antibiotic resistance is one of the most challenging global health threats in our society. Antimicrobial peptides (AMPs) represent promising alternatives to conventional antibiotics for the treatment of drug-resistant infections. However, they are limited by their high manufacturing cost. Engineering living organisms represents a promising approach to produce such molecules in an inexpensive manner. Here, we genetically modified the yeast Pichia pastoris to produce the prototypical AMP apidaecin Ia using a fusion protein approach that leverages the beneficial properties ( e.g., stability) of human serum albumin. The peptide was successfully isolated from the fusion protein construct, purified, and demonstrated to have bioactivity against Escherichia coli. To demonstrate this approach as a manufacturing solution to AMPs, we scaled-up production in bioreactors to generate high AMP yields. We envision that this system could lead to improved AMP biomanufacturing platforms.

Knockdown of REGγ inhibits the proliferation and migration and promotes the apoptosis of multiple myeloma cells by downregulating NF-κB signal pathway.

OBJECTIVES: This study aimed to evaluate the effects of REGγ knockdown on the proliferation, apoptosis and migration of multiple myeloma (MM) cells, and reveal the potential regulatory mechanisms. METHODS: The expression of REGγ on myeloma cells of 28 MM patients was detected by Western blot. shRNA-REGγ-1 and shRNA-REGγ-2 were constructed to downregulate REGγ in RPMI-8226 cells. The proliferation, apoptosis and migration of transfected cells were analyzed by Cell Counting Kit 8 (CCK8), flow cytometry and transwell chamber, respectively. The expression of phosphorylated p65 (p-p65), p65, NF-kappa-B inhibitor ε (IkBε), matrix metalloproteinase 2 (MMP2), B-cell lymphoma xL (Bcl-xL) and X-linked inhibitor of apoptosis protein (XIAP) in transfected cells was detected by Western blot. Using cycloheximide (CHX), the half-life period of IkBε was detected by Western blot. RESULTS: The expression of REGγ was positive in myeloma cells. The proliferation and migration of RPMI-8226 cells were significantly inhibited by shRNA-REGγ-1/shRNA-REGγ-2, while the apoptosis rates were significantly increased (p < 0.05). The expression of p-p65 and IkBε was significantly reduced in RPMI-8226 cells transfected with shRNA-REGγ-1/shRNA-REGγ-2. The degradation of IkBε was significantly lower in RPMI-8226 cells transfected with shRNA-REGγ-1 than the control (longer half-life period). Besides, the expression of MMP2, Bcl-xL and XIAP in RPMI-8226 cells was significantly inhibited by shRNA-REGγ-1/shRNA-REGγ-2. DISCUSSION: Knockdown of REGγ may inhibit the proliferation and migration, and promote the apoptosis of RPMI-8226 cells possibly by downregulating NF-κB signal pathway.

A dual-label time-resolved fluorescence immunoassay for the simultaneous determination of ferritin and ß2 -microglobulin.

BACKGROUND: Lymphocytic leukemia is a kind of primary malignant tumor of hematopoietic tissue. The aim was to establish a dual-label time-resolved fluorescence immunoassay (TRFIA) for the simultaneous determination of ferritin (FER) and ß2 -microglobulin (ß2 -MG) for the early screening and follow-up surveillance of lymphocytic leukemia. METHODS: The sandwich immunoassay was used to detect the concentration of FER, and the competitive immunoassay was used to detect the concentration of ß2 -MG in serum. FER in serum was captured by anti- FER antibody immobilized on microtiter wells, and then banded together with another anti- FER labeled with europium(III) Eu3+ chelate, followed by fluorescence measurement using time-resolved fluorometry (TRF). Sm3+ labeled ß2 -MG and ß2 -MG samples were added to compete with a certain amount of anti-ß2 -MG antibody, followed by fluorescence measurement using TRF. The performance of this dual-label TRFIA was evaluated using the clinical blood and compared with the commercial assays. RESULTS: The linear correlation coefficient (R2 ) of the FER and ß2 -MG standard curves were 0.9914 and 0.9927, respectively. The sensitivity for FER detection was 8 ng/mL (dynamic range 0-1000 ng/mL), the average recovery was 100.51%; The sensitivity for ß2 -MG detection was 1 ng/mL (dynamic range 0-1000 ng/mL), the average recovery was 101.02%. High correlation coefficients (R2 ) were obtained between the commercial assays (R2 =.9966 for FER, and R2 =.9897 for ß2 -MG). CONCLUSION: The present dual-label TRFIA has high sensitivity, specificity, and accuracy in clinical sample analysis. It is an effective detection method for the early screening and follow-up surveillance of the acute and chronic lymphocytic leukemia.

Synthesis and evaluation of multifunctional ferulic and caffeic acid dimers for Alzheimer's disease.

In this study, a series of novel ferulic and caffeic acid dimers was designed and synthesised, and their multifunctional properties against Alzheimer's disease (AD) were evaluated. Results showed that our multifunctional strategy was great supported by enhancing the inhibition of Aß1-42 self-induced aggregation. Moreover, 7b also had potent protective effects against glutamate-induced cell death without significant cell toxicity in mouse hippocampal neuronal HT22 cells and 10c effectively scavenged diphenylpicrylhydrazyl free radicals. Collectively, these data strongly encourage further optimisation of 7b as a new hit to develop multifunctional agents for the treatment of AD.