Objective To develop a gas chromatography-tandem mass spectrometry(GC-MS/MS)method for the simultaneous determination of ephedrine and pseudoephedrine in urine.Methods Urine samples containing ephedrine and pseudoephedrine components were extracted with ethyl acetate,centrifuged to collect the supernatant and evaporated to dryness under a nitrogen stream and then derivatized with heptafluorobutyric anhydride 60 μL at 70 ℃ for 30 min,and re-evaporated under nitrogen,and then solubilized with 50 μL of methanol,and then analyzed by GC-MS/MS.Results The method demonstraed excellent linearity for ephedrine(0.05～10 μg/mL,r=0.999 8)and pseudoephedrine(0.02～5 μg/mL,r=0.999 5).Extraction recoveries ranged from 89.4％～95.8％(ephedrine)and 90.3％～93.8％(pseudoephedrine).Limits of detection and quantification of ephedrine and pseudoephedrine were 0.005 μg/mL and 0.01 μg/mL,the intra-day precision and accuracy were less than 5.87％and 9.56％,respectively,and the inter-day precision and accuracy were less than 7.54％and 9.27％,respectively.The stability of ephedrine and pseudoephedrine in urine in 15 d was good under the conditions of room temperature and-20 ℃.Conclusion The GC-MS/MS analytical method for the analysis of ephedrine and pseudoephedrine components in urine established in this study is accurate,stable and sensitive,which can provide data technical support for the forensic toxicological analysis of amphetamine-type drugs or new psychoactive substances in the cathinone group.

Objective To develop a gas chromatography-tandem mass spectrometry(GC-MS/MS)method for the simultaneous determination of ephedrine and pseudoephedrine in urine.Methods Urine samples containing ephedrine and pseudoephedrine components were extracted with ethyl acetate,centrifuged to collect the supernatant and evaporated to dryness under a nitrogen stream and then derivatized with heptafluorobutyric anhydride 60 μL at 70 ℃ for 30 min,and re-evaporated under nitrogen,and then solubilized with 50 μL of methanol,and then analyzed by GC-MS/MS.Results The method demonstraed excellent linearity for ephedrine(0.05～10 μg/mL,r=0.999 8)and pseudoephedrine(0.02～5 μg/mL,r=0.999 5).Extraction recoveries ranged from 89.4％～95.8％(ephedrine)and 90.3％～93.8％(pseudoephedrine).Limits of detection and quantification of ephedrine and pseudoephedrine were 0.005 μg/mL and 0.01 μg/mL,the intra-day precision and accuracy were less than 5.87％and 9.56％,respectively,and the inter-day precision and accuracy were less than 7.54％and 9.27％,respectively.The stability of ephedrine and pseudoephedrine in urine in 15 d was good under the conditions of room temperature and-20 ℃.Conclusion The GC-MS/MS analytical method for the analysis of ephedrine and pseudoephedrine components in urine established in this study is accurate,stable and sensitive,which can provide data technical support for the forensic toxicological analysis of amphetamine-type drugs or new psychoactive substances in the cathinone group.

Chemoresistance to 5-fluorouracil (5-FU) is a significant challenge in treating colorectal cancer (CRC). Novel combined regimens to thwart chemoresistance are therefore urgently needed. Herein, we demonstrated that the combination of Avenanthramide A (AVN A) and 5-FU has significant therapeutic advantages against CRC. Mechanistically, AVN A directly binds to the S198 site of the histone lysine demethylase KDM4C to promote its degradation, which subsequently fosters H3K9me3 occupancy on the MIR17HG promoter to block its transcription and derepress Bim expression. AVN A enhanced the therapeutic efficacy of 5-FU via impairing the KDM4C/MIR17HG/GSK-3β negative feedback loop. Importantly, the clinical correlation of the KDM4C/MIR17HG/Bim signaling axis with 5-FU response was validated in the refractory CRC patients. We provide evidence for the enhanced effectiveness of 5-FU when combined with AVN A in chemoresistant xenografts, CRC organoids, and Apc ^Min/+ mouse model. Additionally, AVN A mitigated the systemic adverse effects of 5-FU. Overall, our findings demonstrate that combinatorial therapy with AVN A and 5-FU represents an appealing opportunity and highlights KDM4C/MIR17HG/GSK-3β negative feedback loop which confers therapeutically exploitable vulnerability to chemo-refractory CRC patients.

Objective To investigate the application value of neutrophil count(N),neutrophil percentage(N％),neutrophil to lymphocyte ratio(NLR)and neutrophil to albumin ratio(NAR)in the evaluation of ul-cerative colitis(UC)patients.Methods A total of 87 UC patients admitted to the hospital from May 2022 to July 2023 were collected as the UC group,and 42 healthy people in the same hospital during the same period were selected as the control group.Blood routine and biochemical routine related inflammatory indicators in-cluding lymphocyte count(L),N,lymphocyte percentage(L％),N％,C-reactive protein(CRP),erythrocyte sedimentation rate(ESR),albumin(ALB)were collected,NLR and NAR were calculated.The clinical data of UC patients were collected.According to medical history,modified Mayo scoring system and Montreal classifi-cation criteria,UC patients were divided into clinical types,disease activity and lesion extent.The levels of N,N％,NLR and NAR in the control group and UC group and different subgroups of UC were evaluated.Spearman correlation analysis was used to analyze the correlation between the four indexes and ESR and CRP.The receiver operating characteristic curve was drawn to test the diagnostic efficacy of the four indicators a-lone and in combination in the detection of UC patients,and the best cut-off value,sensitivity and specificity were calculated.Results The levels of N,N％,NLR and NAR in UC group were higher than those in control group,and the differences were statistically significant(P＜0.05).There were significant differences in N,N％,NLR and NAR levels between UC patients with different activity and lesion range(P＜0.05).Spearman correlation analysis showed that CRP level was positively correlated with N,N％,NLR and NAR in UC pa-tients(r=0.392,0.343,0.354,0.503,P＜0.001).ESR level was positively correlated with N,NLR and NAR(r=0.383,0.233,0.475,P＜0.05).The area under the curve of N,N％,NLR and NAR alone and in combi-nation were 0.668,0.702,0.723,0.741 and 0.882,respectively.The sensitivity and specificity of the combina-tion of the four were higher(80.5％and 88.1％,respectively).Conclusion N,N％,NLR and NAR have cer-tain clinical value in the diagnosis,disease activity and lesion extent prediction of UC patients.

Purpose: Previous report from the ASCEND-8 trial showed consistent efficacy with less gastrointestinal (GI) toxicity in patients with anaplastic lymphoma kinase-rearranged (ALK+) advanced/metastatic non–small cell lung cancer (NSCLC) treated with ceritinib 450-mg with food compared with 750-mg fasted. In this subgroup analysis, we report outcomes in Asian patients of the ASCEND-8 trial. Materials and Methods: Key efficacy endpoints were blinded independent review committee (BIRC)–assessed overall response rate (ORR) and duration of response (DOR) evaluated per Response Evaluation Criteria in Solid Tumors v1.1. Other efficacy endpoints were investigator-assessed ORR and DOR; BIRC- and investigator-assessed progression-free survival (PFS) and disease control rate; overall survival (OS). Safety was evaluated by frequency and severity of adverse events. Results: At final data cutoff (6 March 2020), 198 treatment-naïve patients were included in efficacy analysis, of which 74 (37%) comprised the Asian subset; 450-mg fed (n=29), 600-mg fed (n=19), and 750-mg fasted (n=26). Baseline characteristics were mostly comparable across study arms. At baseline, more patients in 450-mg fed arm (44.8%) had brain metastases than in 750-mg fasted arm (26.9%). Per BIRC, patients in the 450-mg fed arm had a numerically higher ORR, 24-month DOR rate and 24-month PFS rate than the 750-mg fasted arm. The 36-month OS rate was 93.1% in 450-mg fed arm and 70.9% in 750-mg fasted arm. Any-grade GI toxicity occurred in 82.8% and 96.2% of patients in the 450-mg fed and 750-mg fasted arms, respectively. Conclusion Asian patients with ALK+ advanced/metastatic NSCLC treated with ceritinib 450-mg fed showed numerically higher efficacy and lower GI toxicity than 750-mg fasted patients.

Objective:To analyze the peripheral retinal fluorescence characteristics of normal fundus on ultra-wide field fluorescein angiography (UWFA) images.Methods:A cross-sectional study was conducted.Ninety-five patients (190 eyes) who underwent normal UWFA in Renmin Hospital of Wuhan University from July 2016 to January 2019 were enrolled.There were 94 (49.47%) male eyes and 96 (50.53%) female eyes.Among them, there were 72 (37.89%) mild cataract eyes, 60 (31.58%) moderate and low myopia eyes and 58 (30.53%) subjective blurred vision eyes.The peripheral retinal fluorescence characteristics were divided into vascular-associated feature or non-vascular feature according to whether the retinal vessel involved or not.The subjects were divided into ≤40 years old group and >40 years old group, and the differences in various features between the two groups were compared and analyzed.The study protocol adhered to the Declaration of Helsinki and was approved by an Ethics Committee of Renmin Hospital of Wuhan University (No.WDRY2019-K037). Written informed consent was obtained from each patient prior to any examination.Results:Four non-vascular and five vascular-associated fluorescence features were found in normal peripheral retina.Non-vascular features contained glass hyperfluorescence in 158 (83.16%) eyes, far peripheral retina with mottled florescent band in 82 (43.16%) eyes, granular ground hyperfluorescence in 24 (12.63%) eyes and local mottled fluorescence in 21 (11.05%) eyes.Vascular-associated fluorescence features included peripheral avascular area in 92 (48.42%) eyes, vessels passing through the ora serrata in 66 (34.74%) eyes, microaneurysm in 60 (31.58%) eyes, slight leakage within 10 minutes after angiography in 56 (29.47%) eyes and angiotelectasis in 30 (15.79%) eyes.There were 19.61% (20/102) of eyes with peripheral retinal vessels passing through the ora serrata and 43.10% (44/102) of eyes with microaneurysm and 19.61% (20/102) of eyes with angiotelectasis in >40 years old group, and there were 52.27% (46/88), 18.23% (16/88) and 11.36% (10/88) correspondingly in ≤40 years old group, and the differences were statistically significant ( χ2=22.235, 10.451, 9.259; all at P<0.01). Conclusions:UWFA reveals four non-vascular and five vascular-associated fluorescence characteristics of normal fundus and age might be associated with the distribution of microaneurysm and angiotelectasis.

Objective:To investigate the features of multicolor imaging in the macular region of central serous chorioretinopathy (CSC) patients.Methods:A cross-sectional study was conducted.Thirty-four acute CSC patients (34 eyes) treated in Renmin Hospital of Wuhan University from August 2017 to January 2018 were enrolled.Among the 34 subjects, there were 21 males (21 eyes) and 13 females (13 eyes). The subjects were 26 to 61 years old, with an average age of (37.41±9.35) years.The course of the disease was 5 to 45 days, with an average course of (12.00±2.29) days.All the subjects were examined by color fundus photography, fluorescein angiography (FFA), indocyanine green angiography (ICGA), multicolor imaging, spectral-domain optical coherence tomography (SD-OCT). The image features of each patient were compared and analyzed.The diagnostic accordance rate for leakage point and serous retinal neuroepithelial detachment of multicolor imaging and color fundus photography was calculated according to FFA/ICGA and OCT.This study protocol adhered to the Declaration of Helsinki and was approved by an Ethics Committee of Renmin Hospital of Wuhan University (No.WDRY2019-K037).Results:The serous retinal detachment region showed green light reflection area with clear boundary in 33 eyes (97.06%) in the standard as well as blue and green enhanced multicolor image, with not clear boundary in 1 eye (2.94%). The serous retinal detachment region showed weakly reflective area in 17 eyes (50%) in blue reflectance image, showed weak reflection with clear boundary in 32 eyes (94.11%) in green reflectance image, showed weakly reflection with clear boundary in 33 eyes (97.06%) in infrared reflectance image.The fluorescein leakage point in FFA image was found micro retinal pigment epithelium detachment (PED) in 19 eyes (55.88%), rough light band of retinal pigment epithelium (RPE) in 12 eyes (35.29%), and large PED in 3 eyes (8.82%) in SD-OCT image.The RPE leakage showed red mottled changes in the area of neuroepithelial detachment in 29 eyes (85.29%) in the standard as well as blue and green enhanced multicolor images, presented strong reflection spots in blue reflectance images in 2 eyes (5.88%), showed strong reflective spots in green reflectance in 5 eyes (14.70%), showed strong reflection spot in the weakly reflective area in 33 eyes (97.06%) in infrared reflectance images.Taking FFA/ICGA and OCT as the gold standard, the diagnostic accordance rate of standard multicolor, blue and green enhanced multicolor and infrared reflectance images for serous retinal neuroepithelial detachment and leakage points was higher than that of color fundus photography, and the differences were statistically significant (all at P<0.05). Conclusions:Standard multicolor, blue and green enhanced multicolor and infrared reflectance images can reflect the leakage point and retinal neuroepithelial detachment of acute CSC.Green reflectance image can show serous retinal neuroepithelial detachment of acute CSC.Multicolor imaging can be used as the auxiliary diagnosis method of acute CSC.

Objective:To explore the application of multispectral fundus imaging (MSI) in central serous chorioretinopathy (CSC).Methods:A diagnostic test study was conducted.Eighty-five eyes from 69 patients with macular diseases were enrolled in Renmin Hospital of Wuhan University from September 2017 to May 2018.There were 62 males (77 eyes) and 7 females (8 eyes) among the 69 patients with an average age of (48.0±11.2) years.The patients were divided into CSC group (45 cases, 61 eyes) and non-CSC group (24 cases, 24 eyes) according to whether suffering from CSC or not.Color fundus photography, fundus fluorescein angiography (FFA), indocyanine green angiography, spectral domain-optical coherence tomography and MSI were performed in the two groups and the images were collected.With FFA as the gold standard, the sensitivity, specificity, false positive rate, false negative rate, positive predictive value and negative predictive value of MSI in the diagnosis of CSC were calculated.Kappa consistency test was used to evaluate the consistency of MSI and FFA in diagnosing CSC, and to evaluate the diagnostic value of MSI in CSC.This study protocol adhered to the Declaration of Helsinki and was approved by an Ethics Committee of Renmin Hospital of Wuhan University (No.WDRY2019-K037). Written informed consent was obtained from each subject prior to any medical examination.Results:The sensitivity, specicity, false positive rate, false negative rate, positive predictive value and negative predictive value of MSI for identifying CSC were 88.5%, 91.7%, 8.3%, 11.5%, 96.4% and 75.9%, respectively.The Kappa coefficient of agreement was 0.754.There was no significant difference in the identification of CSC between MSI and FFA ( χ2=1.780, P=0.180). Hyper-reflective signals corresponding to the leakage points were better detected with MSI images at 620-850 nm wavelength.The clear boundary of serous neuroepithelial detachment area was better detected with MSI images at 590-810 nm wavelength.The retinal pigment epithelium damage was better detected with MSI images at 660-850 nm wavelength. Conclusions:There is a good consistency between MSI and FFA.MSI can be an effective noninvasive inspection method for CSC.

Objective To observe the imaging features of cystoid macular edema (CME) in multicolor imaging (MC),and to evaluate the value of MC in the diagnosis of CME.Methods Descriptive case series study.From August 2017 to June 2018,42 eyes of 37 patients with CME diagnosed in the people's Hospital of Wuhan University were included in the study.Among them,there were 24 males and 13 females,with an average age of 48.51 ± 10.29 years.There were 14 eyes with diabetic retinopathy,14 eyes with central retinal vein occlusion,8 eyes with branch retinal vein occlusion,4 eyes with uveitis,and 2 eyes with Eales disease.The macular color fundus photography (CFP) was performed with Visucam 200 non-mydriatic fundus camera of Zeiss company in Germany.MC,frequnce domainoptical OCT (SD-OCT) and FFA were examined by Spectralis HRA2 + OCT of Heidelberg company in Germany.According to the MC standard method,five images,including 488 nm blue reflection (BR),515 nm green reflection (GR),820 nm infrared reflection (IR) imaging and standard MC and blue-green enhancement (BG),were obtained at the same time.Compared with SD-OCT,CFP and MC images were scored.Friedman M test and Wilcoxon signed rank test were used for statistical analysis.Results The standard MC and BG images showed blue-green uplift area or petal-shaped appearance,surrounded by green reflection areas with clear boundaries.BR image can be seen in the low reflexes area.On the GR image,there were patches or cystic low reflection areas,surrounded by a slightly high reflection.On the IR image,patches or cystoid high reflexes can be seen,surrounded by low reflection dark areas with clear boundaries.The average scores of CFP,standard MC,GB,IR,GR and BR were 1.20± 0.94,3.05± 0.99,2.90± 1.04,2.55± 1.27,2.00± 0.94,0.51 ± 0.85 respectively,and the differences were statistically significant (χ2=151.61,P=0.000).The score of CFP were significantly lower than that of standard MC (Z=-5.421),BG (Z=-5.354),IR (Z=-4.714),GR (Z=-4.438) and higher than that of BR (Z=-3.435).The differences were statistically significant (P=0.000,0.000,0.000,0.000,0.001).Conclusions The quality of MC imaging is better than that of CFP.Combined with SD-OCT,it can be used as an assistant method to diagnose CME.

Chinese Medicine Questionnaire (CCMQ-J) consists of sixty independent questionnaires and 9 physical constitutions called subscales. One type is balanced constitution (i.e., gentleness), and the following eight types represent unbalanced constitution: Qi-deficiency constitution, Yang-deficiency constitution, Yin-deficiency constitution, Phlegm-dampness constitution, Damp-heat constitution, Stagnant Blood constitution, Stagnant Qi constitution, and Inherited Special constitution. In this study, we proposed to determine optimal number of groups in 851 participants recruited from crowdsourcing answered CCMQ-J questionnaire consisting of 60 questions. In the present study, we applied k-means clustering with gap statistics to the questionnaire data and the number of optimal groups was estimated by five. The five groups are mainly characterized by 3 subscales in CCMQ-J, i.e. (i) two subscales corresponding to Yang-deficiency and Qi-depress, (ii) three subscales corresponding to gentleness, Yang-deficiency and Qi-depress (iii) Yang-deficiency, (iv) gentleness, and (v) Qi-depress. In the crowdsourcing survey, two subscales, Yang-deficient and Qi-depress are the most frequently occurred in current Japanese people.