Objective: To systematically analyze the confirmatory positivity of different combinations of HBsAg screening results in blood testing, providing data to support the optimization of blood donor eligibility management. Methods: A retrospective analysis was conducted on blood screening data from 174 266 voluntary blood donor samples at the Chongqing Blood Center between October 2021 and September 2022. Samples with inconsistent results between the two HBsAg enzymelinked immunosorbent assays (ELISA) and individual donor nucleic acid testing (NAT) were confirmed using an electrochemiluminescence immunoassay (ECLIA) and a neutralization test. The detection efficacy of four different HBsAg ELISA reagents was compared using the HBsAg-confirmed positive samples. Results: A total of 767(0.44%) HBV-reactive (HB-sAg and/or HBV DNA reactive) samples were detected. Among them, 344 samples with discordant serological and NAT results were collected, of which 64(18.6%) were confirmed positive by neutralization test. Additionally, 5 samples that were neutralization-negative but double-reactive for HBsAg and HBV DNA were confirmed as positive according to FDA guidance, resulting in a total of 69(20.1%) confirmed HBsAg-positive samples. There were significant differences in the neutralization test confirmation rates among different screening result categories (P<0.05): The group with dual HBsAg reagent reactivity (double reactive) & NAT-negative had the highest confirmation rate (96.9%, 31/32); the group reactive to only reagent 2 (single reactive) had a rate of 25.7% (29/113); while the confirmation rates for samples reactive to only reagent 1 and samples with isolated HBV DNA positivity were extremely low [0(0/34) and 2.4%(4/165), respectively]. The four commercial reagents showed significant differences in their ability to detect confirmed positive samples that were initially single reactive (P<0.05). Conclusion: Given the performance variations among HBsAg screening reagents, thorough performance verification is essential before implementation. When NAT is negative, dual HBsAg reactivity in screening can serve as a basis for confirming infection and directly deferring blood donors. However, confirming infection in donors with single HBsAg reactivity is more challenging, necessitating supplementary tests to rule out infection risk.

Objective To investigate the effect of different radiotherapy doses on the prognosis of patients with stage cT1-4N0M0 esophageal squamous cell carcinoma(ESCC)who received radical radio(chemo)therapy categorized into subgroups with different tumor local factors.Methods A retrospective analysis was conducted on 256 patients with clinically nonmetastatic esophageal squamous cell carcinoma.The optimal cutoff for tumor local factors was determined.The relationship between latest treatment efficacy and tumor local factors was analyzed,and independent indicators affecting patient overall survival(OS)were examined using multivariate analysis.The subgroup analysis was performed to determine the correlation between selected factors and radiation therapy doses.Results The shorter the X-ray length of esophageal tumor lesion and the smaller the lesion canal wall thickness and gross tumor volume(GTV),the better the latest treatment efficacy of the patients(χ2=9.066,10.310,15.661,respectively,P=0.011,0.006,P＜0.001).Multivariate analysis results showed that GTV(P＜0.001),radiation dose(P=0.038),and latest treatment efficacy(P＜0.001)were independent predictors of the patients'OS,and the latter two were also independent predictors of the patients'progression-free survival(PFS)(P=0.033,＜0.001).Subgroup analysis further showed that high doses of radiotherapy(over 60 Gy)resulted in good OS(χ2=5.040,4.588,5.400,P=0.025,0.032,0.020)and PFS(χ2=6.089,4.353,6.459,P=0.014,0.037,0.011)in the subgroup with maximum wall thickness below 3.7 cm,with esophageal lesions with GTV below 37.34 cm3,or not receiving simultaneous chemotherapy.Conclusion Local tumor factors are important prognostic factors of ESCC patients treated with radical radio(chemo)therapy.Patients with thin lesion walls and small tumor volumes may greatly benefit from high doses of radiation(over 60 Gy).

Liquiritigenin(LG)is a flavone of pharmacological importance,however,its application potential is severely limited due to its poor water solubility.LG could be disassociated slightly in water to form phenolate anion,therefore,better solubilization effect is expected by inclusion with cationic cyclodextrins(CCDs).In this work,four kinds of CCDs modified with amino groups at the primary face were synthesized,and their solid inclusion complexes with LG were successfully prepared by preparing their saturated solutions.The formation of the solid inclusion complexes was confirmed by scanning electron microscopy(SEM)and powder X-ray diffraction(PXRD),and their supramolecular binding behavior in solution was studied using multiple techniques.A 1∶1 inclusion stoichiometry of inclusion complexation was defined using Job plot by ultraviolet-visible(UV-vis)spectroscopy,and their binding stability constants(Ks)were determined as 2862.77,3494.70,6521.85 and 9599.48 L/mol using UV-vis spectroscopic titration,far more superior to that of nativeβ-CD(Ks=236.79 L/mol).This indicated that the amino side chains on CCDs could actively participate in the inclusion complexation through anion-cation interactions,significantly strengthening the host-guest binding between CCDs and LG.The inclusion modes were further elucidated based on proton and two-dimensional rotating-frame overhauser enhancement spectroscopy(2D-ROESY)nuclear magnetic resonance(NMR)experiments and molecular docking.Water solubility of LG was dramatically promoted up to 4.9 mg/mL,which was 70-fold higher than that of native LG.This study could draw inspiration for the binding and solubilization of phenols such as flavones by design of cationic macrocyclic molecules.

Haloacetic acids(HAAs),as a class of disinfection byproducts in drinking water,pose potential threats to human health,so the rapid,accurate and simultaneous detection of HAAs is of great significance for ensuring drinking water safety.Aiming at the challenges in HAAs detection and risk analysis,a novel method for synchronous rapid detection of seven kinds of HAAs in drinking water based on in situ derivatization technology and headspace gas chromatography was developed in this study.Through single-factor optimization experiments,the optimal reaction parameters for in situ derivatization were determined,including the type and dosage of salting-out agent,the acidity of reaction system,the amount of phase transfer catalyst,the dosage of derivatization agent,and the extraction solvent volume.Methodologic validation showed that the seven kinds of HAAs exhibited excellent linear relationships within their respective detection concentration ranges(R2>0.998).The method detection limits(MDLs)ranged from 0.04 to 0.33 μg/L,and the limits of quantification(LOQs)were between 0.14 and 1.34 μg/L.For real water samples,the average spiked recoveries of the seven HAAs ranged from 90.9%to 107.7%,with relative standard deviation(RSDs)between 1.55%and 6.49%,and the HAAs contents in all tested samples were below the limits specified in the Standards for Drinking Water Quality(GB 5749-2022)of China.This method was featured with simple operation,fast analysis speed,high sensitivity,and good accuracy,providing an efficient and reliable technical support for routine monitoring of HAAs contaminants in drinking water and showing promising application value for widespread promotion.

BACKGROUND:Enhancing the differentiation of induced pluripotent stem cells into lung stem cells is crucial for repairing lung injuries.NKX2.1 is the earliest marker of lung epithelial differentiation and plays a significant regulatory role in lung development.However,the impact of its expression on the differentiation of induced pluripotent stem cells into lung stem cells remains inadequately understood.OBJECTIVE:To investigate the effect of NKX2.1 on the differentiation of induced pluripotent stem cells into lung stem cells.METHODS:Induced pluripotent stem cells were cultured in vitro.The expression of specific pluripotent stem cell genes was assessed using real-time fluorescence quantitative PCR.NKX2.1 was overexpressed in induced pluripotent stem cells,which were then induced to differentiate into lung stem cells.The expression of FoxA2,SOX9,and P63 was determined via quantitative PCR and immunofluorescence on day 7 of induction of differentiation.The expression of the alveolar marker SPB and SPC was evaluated through immunofluorescence staining on day 7 of induction of differentiation.RESULTS AND CONCLUSION:(1)Induced pluripotent stem cells in vitro were tightly packed and showed typical clonoid growth and significantly expressed stem cell-specific genes OCT-4,SOX2,and NANOG.(2)Compared with the non-transfected control group,the expression of NKX2.1 in human induced pluripotent stem cells was significantly increased in the NKX2.1 overexpression group(P＜0.000 1).(3)Seven days after induction of differentiation,compared with the non-transfected control group,the expression of lung stem cell-related markers FoxA2,SOX9,and P63 was significantly increased in the NKX2.1 overexpression group(P＜0.000 1).(4)Thirteen days after induction of differentiation,compared with the non-transfected control group,the fluorescence intensity of alveolar cell marker molecules SPB and SPC increased significantly in the overexpression NKX2.1 group.The results show that NKX2.1 can promote the differentiation of induced pluripotent stem cells into lung stem cells.

BACKGROUND:Enhancing the differentiation of induced pluripotent stem cells into lung stem cells is crucial for repairing lung injuries.NKX2.1 is the earliest marker of lung epithelial differentiation and plays a significant regulatory role in lung development.However,the impact of its expression on the differentiation of induced pluripotent stem cells into lung stem cells remains inadequately understood.OBJECTIVE:To investigate the effect of NKX2.1 on the differentiation of induced pluripotent stem cells into lung stem cells.METHODS:Induced pluripotent stem cells were cultured in vitro.The expression of specific pluripotent stem cell genes was assessed using real-time fluorescence quantitative PCR.NKX2.1 was overexpressed in induced pluripotent stem cells,which were then induced to differentiate into lung stem cells.The expression of FoxA2,SOX9,and P63 was determined via quantitative PCR and immunofluorescence on day 7 of induction of differentiation.The expression of the alveolar marker SPB and SPC was evaluated through immunofluorescence staining on day 7 of induction of differentiation.RESULTS AND CONCLUSION:(1)Induced pluripotent stem cells in vitro were tightly packed and showed typical clonoid growth and significantly expressed stem cell-specific genes OCT-4,SOX2,and NANOG.(2)Compared with the non-transfected control group,the expression of NKX2.1 in human induced pluripotent stem cells was significantly increased in the NKX2.1 overexpression group(P＜0.000 1).(3)Seven days after induction of differentiation,compared with the non-transfected control group,the expression of lung stem cell-related markers FoxA2,SOX9,and P63 was significantly increased in the NKX2.1 overexpression group(P＜0.000 1).(4)Thirteen days after induction of differentiation,compared with the non-transfected control group,the fluorescence intensity of alveolar cell marker molecules SPB and SPC increased significantly in the overexpression NKX2.1 group.The results show that NKX2.1 can promote the differentiation of induced pluripotent stem cells into lung stem cells.

This study aims to investigate the effect of Lactiplantibacillus plantarum LP12 on obe-sity prevention.In our study,Lactiplantibacillus plantarum LP12 was added to the diet for feed-ing,and the blood biochemistry status of rabbit,as well as the antioxidant effect of serum and liver samples were analyzed by determining the body weight change and feed intake of Japanese White rabbits.The changes in colony structure and abundance were also analyzed by 16S rDNA sequen-cing.The results showed that supplementation of Lactiplantibacillus plantarum LP12 inhibits weight gain,decreases serum glucose and ALT levels,and increases SOD activity in the liver.16S RNA gene sequencing analysis showed that the addition of Lactiplantibacillus plantarum LP12 increases the abundance of Bacteroidetes and Desulfovibrioides at the phylum level,and the supple-mentation of Lactiplantibacillus plantarum LP12 increases the abundance of Muribaculaceae at the genus level.Predictive analysis of microbiota function revealed that the supplementation of Lactiplantibacillus plantarum LP12 positively regulated iron-sulfur clusters and Zn-dependent proteases.In conclusion,the addition of Lactiplantibacillus plantarum effectively inhibits weight gain in Japanese White rabbits,enhances the antioxidative activity of the liver,and induces altera-tions in the gut microbiota composition of these rabbits.These findings lay an experimental foun-dation for further exploring the mechanisms by which Lactobacillus plantarum LP12 exerts its preventive effects against obesity and promotes metabolic health.

AIM To study the chemical constituents from salt-processed Litchi Semen and their antioxidant activities.METHODS The 85％ethanol extract from salt-processed Litchi Semen was isolated and purified by silica gel,Sephadex LH-20,MCI,ODS and semi-preparative HPLC,then the structures of obtained compounds were identified by physicochemical properties and spectral data.DPPH and ABTS+free radical scavenging method were used to evaluate their antioxidant activities.RESULTS Fifteen compounds were isolated and identified as dehydrocostuslactone(1),ananosmoside A(2),funingensin A(3),(2S)-pinocembrin-7-O-(6-O-α-L-rhamnopyranosyl-β-D-glucopyranoside)(4),liquiritienin(5),quercetin(6),rutin(7),isorhamnetin-3-O-β-rutinoside(8),procyanidin A2(9),procyanidin A1(10),ethyl protocatechuate(11),5-hydroxymethylfurfural(12),di(2-ethyl-hexyl)phthalate(13),nicotinamide(14),(10E,15Z)-9,12,13-trihydroxyoctadeca-10,15-dienoic acid(15).Compounds 6-7,9-10 exhibited scavenging activities against DPPH radicals with IC50 values of(12.929±1.232),(14.104±0.946),(10.417±1.736),(6.944±0.030)μmol/L,respectively.Compounds 6-10 exhibited scavenging activities against ABTS+radicals with IC50 values of(21.952±0.577),(25.683±0.625),(22.970±1.336),(20.210±1.435),(18.725±0.324)μmol/L,respectively.CONCLUSION Compounds 1,5,14-15 are isolated from Litchi genus for the first time.Compounds 6-7,9-10 have strong in vitro antioxidant activities.

Sodium-glucose co-transporter 2(SGLT2)inhibitors are a new class of oral hypoglyce-mic drugs with definite hypoglycemic effects,low risk of hypoglycemia,cardiovascular protection,and kidney benefits.In recent years,SGLT2 inhibi-tors have been widely used in clinical practice,and their interactions with other drugs have gradually attracted attention.The SGLT2 inhibitors commonly used in China's clinic include canagliflozin,dapa-gliflozin,empagliflozin,ertugliflozin and hena-gliflozin currently,they are mainly metabolized by the phase Ⅱ metabolic enzyme uridine diphos-phate glucuronosyltransferase(UGT),and various transporters are involved in the disposal of SGLT2 inhibitors in vivo.This article reviews the pharma-cokinetic characteristics of different SGLT2 inhibi-tors mentioned above,as well as their pharmacoki-netic interaction studies with various drugs such as statins,antineoplastic drugs,antimicrobials,nonste-roidal anti-inflammatory drugs and traditional Chi-nese medicine,in order to promote the safe and ra-tional use of SGLT2 inhibitors in clinical practice.

Objective:To explore the molecular mechanism of miR-4508 regulating the inflammatory response of human bronchial epithelial cells induced by representative chrysotile asbestos.Methods:The chrysotile asbestos was ground into ultrafine dust using a horizontal planetary instrument, and human bronchial epithelium (16HBE) cells were taken as the object of infection. Cell survival rate was detected by cell counting kit-8 method, cytotoxicity was detected by lactate dehydrogenase (LDH) kit. The released of inflammatory factor IL-6 was detected by electrochemical luminescence. The released inflammatory factor IL-8 was detected by enzyme-linked immunosorbent assay. The expression level of miR-4508 was screened and verified by reverse transcription-quantitative real-time polymerase chain reaction. After 16HBE cells were treated with AKT inhibitor MK2206, the phosphorylation levels of AKT and PTEN were detected by western blot. The expression levels of AKT and PTEN and the contents of IL-6 and IL-8 were detected in miR-4508 overexpression and interference experiments.Results:With the increase of chrysotile asbestos exposure concentration, the cell survival rate decreased in a concentration-dependent manner, and the LDH content gradually increased. The secretion of IL-6 and IL-8 in chrysotile 25, 50 and 75 μg/ml groups were (325.92±8.61) pg/ml, (331.51±4.96) pg/ml, (378.74±13.77) pg/ml, and (94.95±3.11) pg/ml, (357.60±1.80) pg/ml, (537.19±3.11) pg/ml, respectively, while the group with 0 μg/ml chrysotile was (95.85±1.20) pg/ml and (7.81±0.00) pg/ml ( P＜0.05). In addition, chrysotile asbestos exposure to 16HBE could induce the high expression of miR-4508 . After pretreatment with MK2206, the phosphorylation levels of AKT and PTEN were decreased, the contents of IL-6 and IL-8 were significantly decreased, and the expression level of miR-4508 was significantly reduced. Overexpression of miR-4508 significantly increased the expressions of AKT and PTEN, and the contents of IL-6 and IL-8 ( P＜0.01). After interfering with miR-4508, the expressions of AKT and PTEN were significantly decreased, and the contents of IL-6 and IL-8 were significantly decreased ( P＜0.01). Conclusions:Chrysotile asbestos can induce the inflammatory response of 16HBE cells and up-regulate the expression level of miR-4508. The up-regulation of miR-4508 promotes the 16HBE inflammatory response induced by chrysotile asbestos through the PI3K/AKT pathway.