Objective To observe polyclonal antibodies immobilized on magnetic submicron particles (MSP) as affinity adsorbents and test the reliability of predicted maximum adsorption activity of an enzyme/mutant from a cell lysate (Vs) in recognizing positive mutants.Methods Escherichia coli alkaline phosphatase (ECAP) and Pseudomonas Aeruginosa arylsulfatase (PAAS) were purified by affinity chromatography to serve as immunogens for the preparation of their antisera, which after fractionation by 33％ ammonium sulfate and DEAE-cellulose chromatography yielded the respective polyclonal antibodies.After activation of COOH on MSP, polyclonal antibodies of each enzyme were immobilized to give MSP-polyAb.Activities of an adsorbed enzyme were measured with a chromogenic substrate of 4-nitrphenol by determining absorbance at 405nm after the termination of reaction by alkali.Based on the response curve of activities of the adsorbed enzyme to protein quantities of a lysate, Vs was predicted for comparison.Results The maximum adsorption quantity of ECAP or PAAS on the respective MSP-polyAb was about 2.0mg/g.Specific activity of ECAP after affinity purification was about 70-fold of that of PAAS.ECAP mutant R168K showing about 50％ activity improvement versus ECAP was recognized by comparison of Vs predicted with only 2.5μg of MSP-polyAb;with PAAS mutant G138S as the starting one, the use of 10.0μg of MSP-polyAb to predict Vs recognized the mutants bearing more than 20％ activity improvement.Conclusion With an optimized quantity of MSP-polyAb to predict Vs, weak positive mutants of an enzyme of low activity can be recognized when activities of the adsorbed enzyme/mutant are reliably measured.

Objective : To investigate the role of squalene epoxidase (SQLE) knockout in anti-tumor effect vial im- proving CD8 + T cell infiltration in melanoma tumor microenvironment． Methods : Both immunodeficient and immu- nocompetent mice were inoculated with SQLE knockout B16F10 cells to determine the cell-autonomous and non-au- tonomous regulation of malignancy．Antibody blockade ，Luminex multiplex assays ，and flow cytometry were em- ployed to explore the impact of SQLE gene knockout on the secretion of cytokines / chemokines and immune cell in- filtration．Bioinformatics analysis was conducted to validate the correlation between SQLE expression and immune infiltration as well as clinical prognosis in melanoma patients. Results : Compared with immunodeficient mice， SQLE knockout significantly inhibited melanoma proliferation in immunocompetent mice and prolonged their surviv- al．SQLE knockout induced the secretion of cytokines and chemokines from tumor cells，improved CD8 + T cell in- filtration in the tumor microenvironment，thereby potentiating anti-tumor immunity．Bioinformatics analysis sugges- ted a significant correlation between SQLE and its corresponding immune infiltration markers with the prognosis of melanoma patients． Conclusion SQLE regulates anti-tumor immunity by controlling cytokines and chemokines re- leasing in tumor microenvironment，thus holding promise as a novel tumor immunotherapy target and efficacy predic- tion molecular indicator.

BACKGROUNDOveractive bladder (OAB) is a series of symptoms with high prevalence in elderly people. This study was conducted using the overactive bladder symptom score (OABSS) to evaluate the efficacy of solifenacin succinate for the treatment of OAB.

METHODSThis was a prospective, multicenter, single-arm, 12-week study that enrolled 241 OAB patients. The patients received 5-10 mg/day solifenacin. Changes in OABSS, symptoms from voiding diary, perception of bladder condition (PPBC) score, international prostate symptom score (IPSS) and quality of life (QOL) were evaluated at weeks 0, 4, and 12. The relationship between OABSS and PPBC score or parameters of voiding diary was also evaluated.

RESULTSAt baseline, the mean OABSS for all patients was 9.41 ± 2.40, and was reduced significantly at week 12 (-3.76 points; 61.21%, P < 0.0001). The OABSS subscore, PPBC score, IPSS, and QOL were also significantly reduced during the study (P < 0.0001). The overall incidence of adverse events was 19.91% (44 cases). The gastrointestinal system was the most commonly affected (11.31%). Around 5.88% of the cases had adverse events related to the genitourinary system. There was a strong correlation between OABSS and urinary symptoms that was recorded in the 3-day voiding dairy.

CONCLUSIONSWe showed that solifenacin was clinically effective for relieving OAB symptoms, considering the balance between efficacy, patients' well-being, and tolerability. OABSS integrates four OAB symptoms into a single score and can be a useful tool for research and clinical practice.

Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Muscarinic Antagonists ; therapeutic use ; Prospective Studies ; Quality of Life ; Quinuclidines ; therapeutic use ; Solifenacin Succinate ; Tetrahydroisoquinolines ; therapeutic use ; Treatment Outcome ; Urinary Bladder, Overactive ; drug therapy