A method for simultaneous determination of 21 kinds of Aconitum alkaloids(ATS)in biological specimens and infused liquor using ultra-performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)was developed.The biological samples were pretreated with methanol-acetonitrile(1∶2,V/V)for protein precipitation,while infused liquors were diluted 100-fold with acetonitrile,followed by centrifugation,and filtration by a 0.22-μm membrane.Chromatographic separation was carried out on an EC-C18 column using gradient elution with the mixture of 10 mmol/L ammonium acetate and 0.2%formic acid as mobile phase A and acetonitrile as mobile phase B.With this method,all the analytes were separated within 9.5 min.The samples were detected in positive ESI mode with dynamic multiple reaction monitoring(MRM)and quantified via external standard calibration.The results showed that the concentrations of the analytes in the range of 2-1000 ng/mL had excellent linearity(R2>0.9992)with the peak area.The developed method was successfully used for detection of 21 kinds of aconitum alkaloids,with limits of detection of 0.5-2 ng/mL,quantification limits of 2-6 ng/mL,intra/inter-day precision≤6.0%,spiked recoveries of 89.4%-100.9%,extraction recoveries of 74.2%-104.4%,and matrix effects ranging from-11.1%to 9.2%in blood/urine.The method was applied to detection of 12 samples from 4 fatal aconite poisoning cases,and all 21 kinds of ATS with total alkaloid concentrations of 0.04-4.18 μg/mL in blood and 154.96-422.83 μg/mL in medicinal liquors were detected.Tissue distribution revealed that the order of concentrations from highest to lowest is as follows:urine(157.22 μg/mL)>gastric contents(51.37 μg/mL)>kidney(21.6 μg/g)>whole blood(4.18 μg/mL)>liver(0.03 μg/g).This method showed many advantages such as simple pretreatment,low detection limits,accurate quantification,broad analyte coverage,and superior anti-interference capability in complex matrices,proving ideal for forensic and toxicological analysis of aconitum alkaloids.

Objective To investigate the improvement effects of homogeneous fecal microbiota transplantation(FMT)on chemotherapy-induced diarrhea(CID)in mice.Methods Fifteen C57BL/6N mice were divided into control group,CID model group and CID+FMT group according to the random number distribution and remainder grouping method,with 5 mice per group.Control group received no intervention,and their feces were used to prepare fecal bacteria suspension.CID model group was injected intraperitoneally with fluorouracil(65 mg/kg)for 5 consecutive days to construct the CID mouse model,followed by gavage with 0.1 ml of saline on alternate days.CID+FMT group was given 0.1 ml fecal bacteria suspension gavage on alternate days for one week,followed by intraperitoneal injection of fluorouracil(65 mg/kg)for 5 consecutive days to construct the CID mouse model,with the experiment ending on the 14th day.During the experiment,the mice's food intake and body weight were recorded.At the end of the experiment,the mice were euthanized with deep carbon dioxide anesthesia,and the mice colonic specimens from cecum to anus were collected for hematoxylin and eosin(HE)staining and histopathological examination.Fecal samples were collected for 16S rRNA gene sequencing.Shannon index,Simpson index and Chao1 algorithm were used to analyze the α-diversity species of the intestinal flora in each group of mice.Similarity analysis(Anosim)was used to perform non-parametric on the inter-group differences of intestinal flora among the mice.Linear discriminate analysis size effect(LEfSe)and nonmetric multidimensional scaling(NMDS)were employed to analyze the intestinal dominant flora and the similarity classification relationships in each group of mice.Results The colonic specimen's length from cecum to anus in CID model group was significantly shorter than that in control group(P<0.05),while there was no significant difference between CID+FMT group and CID model group(P>0.05).The weight of mice in CID model group decreased by 42.04%,while control group mice gained 10.24%,with a significant difference between the two groups(P<0.05).The weight of mice in CID+FMT group decreased by 8.12%,which was significantly improved compared to CID model group(P<0.05).HE staining results revealed the intestinal mucosal structure in CID model group was severely damaged,with atrophy and deformation,accompanied by inflammatory cell infiltration,and the pathological score was higher than that of control group(P<0.05).Compared with CID model group,the intestinal mucosal integrity and crypt cells in the CID+FMT group were improved,with less damage,and the pathological score was lower than that of CID model group,but the difference was not statistically significant(P>0.05).The α-diversity analysis showed that there were significant differences in the Shannon,Simpson and Chao1 indices among the three groups(P<0.05).ANOSIM and NMDS analysis revealed that the intestinal flora in CID+FMT group was closer to the normal intestinal flora compared to CID model group.LEfSe analysis showed that the intestinal flora in CID model group was enriched in famliy_Bacteroidaceae,and the intestinal flora in CID+FMT group was similar to that of control group,with an enrichenment of familiy_Enterobacteriaceae.Conclusion Homogeneous FMT can improve the abundance of intestinal flora in CID mice,making it more similar to normal intestinal flora,thereby protecting intestinal mucosa,reducing damage and alleviating the severity of CID.

Tuberculosis is a chronic infectious disease caused by Mycobacterium tuberculosis,pri-marily transmitted through respiratory droplets,and poses a significant global public health challenge.Despite the availability of vaccines and chemotherapy regimens,the emergence of drug resistance and high incidence rates continue to render the prevention and control situation severe.The intestinal mi-crobiota modulates host immunity through metabolites such as short-chain fatty acids,influencing macrophage function,T-cell differentiation,and inflammatory responses,thereby regulating tubercu-losis susceptibility,disease progression,and treatment responses.This review systematically summa-rized the role of intestinal microecology in immune regulation of tuberculosis and its potential applica-tion strategies,delved into the possible immune regulatory mechanisms,proposed that the interaction between intestinal microbiota and immune checkpoint inhibitors may serve as a warning for the clinical risk of tuberculosis reactivation,and analyzed the potential of intestinal microecology as a therapeutic target for tuberculosis.

Objective To investigate the role of Sneathia sanguinegens(S.sanguinegens)in the development of unexplained recurrent spontaneous abortion(URSA).Methods A case-control study was conducted to analyze the vaginal flora characteristics of 65 patients with URSA and 18 healthy controls through 16S rRNA gene sequencing.Toxicity profile of S.sanguinegens on human cervical cancer cells(ME-180),human umbilical vein endothelial cells(HUVEC)and human placental choriocarcinoma cells(JEG-3)was analyzed at the cellular level to assess the mechanism of it in adverse pregnancy outcomes.And S.sanguinegens was used to infect C57BL/6J mice to explore the toxic effect on living organisms.Results The relative abundance of Sneathia was increased in patients with URSA compared with healthy controls.It was positively correlated with the number of miscarriages,and was attributed to S.sanguinegens.We also found that S.sanguinegens damaged ME-180,JEG-3 and HUVEC cells.The degree of cellular damage was related to the level of S.sanguinegens added.Intravenous infection with S.sanguinegens caused inflammatory damage in several organs and extramedullary hematopoiesis in the spleen.Conclusion S.sanguinegens is closely related to URSA and should be emphasized in patients with high vaginal bacterial load.

Objective To investigate the role of Sneathia sanguinegens(S.sanguinegens)in the development of unexplained recurrent spontaneous abortion(URSA).Methods A case-control study was conducted to analyze the vaginal flora characteristics of 65 patients with URSA and 18 healthy controls through 16S rRNA gene sequencing.Toxicity profile of S.sanguinegens on human cervical cancer cells(ME-180),human umbilical vein endothelial cells(HUVEC)and human placental choriocarcinoma cells(JEG-3)was analyzed at the cellular level to assess the mechanism of it in adverse pregnancy outcomes.And S.sanguinegens was used to infect C57BL/6J mice to explore the toxic effect on living organisms.Results The relative abundance of Sneathia was increased in patients with URSA compared with healthy controls.It was positively correlated with the number of miscarriages,and was attributed to S.sanguinegens.We also found that S.sanguinegens damaged ME-180,JEG-3 and HUVEC cells.The degree of cellular damage was related to the level of S.sanguinegens added.Intravenous infection with S.sanguinegens caused inflammatory damage in several organs and extramedullary hematopoiesis in the spleen.Conclusion S.sanguinegens is closely related to URSA and should be emphasized in patients with high vaginal bacterial load.

AIM To establish a quantitative analysis of multi-components by single-marker (QAMS) method for the simultaneous content determination of synephrine,syringin,magnoflorine,narirutin,naringoside,hesperridin,neoesperridin,limonin,naringenin,honokiol and magnolol in different compatibility ratios of Fructus Aurantii Immaturus-Magnoliae Officinalis Cortex drug pair.METHODS The ratios of 1:1,1:2 and 2:1 were adopted in the compatibility of Fructus Aurantii Immaturus and Magnoliae Officinalis Cortex,respectively.The analysis was performed on a 30℃ thermostatic Agilent 5TC-C18 column (4.6 mm × 250 mm,5μm),with the mobile phase comprising of methanol-0.1％ phosphoric acid flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelength was set at 280 nm.Hesperidin was used as an internal standard to calculate the relative correction factors of the other ten constituents,after which the content determination was made.RESULTS Eleven constituents showed good linear relationships within their own ranges (r≥0.9990),whose average recoveries were 97.04％-99.45％ with the RSDs of 1.22％-2.48％.The result obtained by QAMS approximated those obtained by external standard method.CONCLUSION This simple,accurate and reproducible method can be used for the quality control of Fructus Aurantii Immaturus-Magnoliae Officinalis Cortex drug pair.The extraction of various constituents is more favourable when the compatibility ratio of two medicinal materials is 1∶1.

Objective To explore the value of serum D-dimer(D-D),fibrinogen(FIB),platelet(PLT),C-reactive pro-tein(CRP)and tissue plasminogen activator inhibitor(PAI)-1 levels in predicting lower extremity deep vein thrombosis(DVT)after hip joint surgery in the elderly.Methods A retrospective analysis was performed on 165 elderly patients with hip joint surgery admitted from February 2020 to May 2022,including 89 males and 76 females,aged from 60 to 75 years old with an average of(66.43±5.48)years,and there were 102 cases of femoral neck fracture and 63 cases of femoral head necrosis.Serum levels of D-D,FIB,PLT,CRP and PAI-1 tests were performed in all patients within 24 hours after admission,and the patients were divided into DVT group and non-DVT group according to whether they developed DVT.Results The levels of D-D,FIB,PLT,CRP,and PAI-1 in the DVT group were higher than those in the non-DVT group(P＜0.001).Spearman analysis showed that DVT was positively correlated with PLT,CRP,D-D,FIB,and PAI-1 levels(r=0.382,0.213,0.410,0.310,0.353,all P＜0.001).The results of binary Logistic regression analysis showed that D-D and PLT were independent factors affecting the occurrence of DVT(OR=0.038,0.960,P=0.032,0.011).The area under curve(AUC)of D-D,FIB,PLT,CRP,PAI-1,and the five combined predictions for DVT were 0.843,0.692,0.871,0.780,0.819,and 0.960,respectively.The AUC of the five combined predictions was higher than that of the single prediction(P＜0.05).Conclusion D-D,FIB,PLT,CRP and PAI-1 are effective in predicting DVT after hip surgery in the elderly,and the combined prediction of the five factors has higher effi-cacy.

Objective To explore the clinical efficacy and safety of one-stage posterior lesion removal and internal spinal fixation in patients with lumbar Brucellosis spondylitis.Methods The clinical data of 24 patients admitted from October 2017 to October 2022 were retrospectively analyzed,2 patients were lost to follow-up at 10 months after surgery,at the final 22 cases were included in the study,including 13 males and 9 females with an average age of(52.00±6.89)years old,were treated with one-stage posterior lesion removal and internal spinal fixation.The operation time,intraoperative bleeding,follow-up time,ery-throcyte sedimentation rate(ESR)and C-reactive protein(CRP)before and after operation were recorded.The pain visual ana-logue scale(VAS),Oswestry disability index(ODI),the Japanese Orthopaedic Association(JOA)score for neurofunction,American Spinal Injury Association(ASIA)spinal cord injury grade and modified MacNab criteria were ussed to evaluate the efficacy.Results All patients were followed up from 12 to 30 months with an average of(17.41±4.45)months.The operation time was 70 to 155 min with an average of(1 16.59±24.32)min;the intraoperative bleeding volume was 120 to 520 ml with an average of(275.00±97.53)ml.CRP and ESR levels decreased more significantly at 1 week and at the final follow-up than pre-operative levels(P＜0.05).VAS,JOA score and ODI at 1 week and at the latest follow-up were more significantly improved than preoperative results(P＜0.05).There was no significant difference between ASIA preoperative and 1 week after operation(P＞0.05),and a significant difference between preoperative and last follow-up(P＜0.05).In the final follow-up,21 patients had ex-cellent efficacy,1 patient had fair,and there was no recurrence during the follow-up.Conclusion One-stage transpedicular le-sion removal and internal spinal fixation,with few incisions and short operation time,helps the recovery of neurological func-tion,and the prognosis meets the clinical requirements,which can effectively control Brucella spondylitis.

Objective To investigate the differential expression of miR-124-3p in peripheral blood and clinical sig-nificance of patients with β-thalassemia.Methods Peripheral blood samples were collected from 33 patients with β-thalassemia and 30 healthy controls in Ningde Municipal Hospital Affiliated to Ningde Normal University from June 2021 to August 2022.The expression level of miR-124-3p was detected.Luciferase reporter gene was used to verify the interaction between miR-124-3p and ERF 3'UTR.The correlation between differential expression of miR-124-3p and β-thalassemia was analyzed and the clinical diagnostic value of miR-124-3p for β-thalassemia was eval-uated.Results Compared with healthy control individuals,the expression of miR-124-3p was significantly up-reg-ulated in patients with β-thalassemia(P＜0.001).The genotype of miR-124-3p high expression group was 84.2%β0(16/19),the genotype of low expression group was 55.6%β+(10/18).ROC curve analysis showed that miR-124-3p had predictive efficacy for β-thalassemia(AUC:0.842).Luciferase reporter gene analysis showed that ERF gene was the regulatory target of miR-124-3p.Conclusions The differential expression of miR-124-3p in patients with β-thalassemia is closely related to the genotype and clinical severity of thalassemia,and ERF is negatively reg-ulated by miR-124-3p.miR-124-3p may be an effective diagnostic biomarker for β-thalassemia.

This study was aimed at understanding the etiological and genetic characteristics of Yersinia enterocolica isolated in Jiangsu Province between 2005 and 2019.All 110 identified strains of Y.enterocolica were from patients with foodborne diar-rhea in Jiangsu Province,or from pigs,dogs,cattle,sheep,poultry,flies,or food.Virulence genes,biological serotypes,drug resistance,multilocus sequence typing(MLST),and core genome multilocus sequence typing(cgMLST)based on whole-genome sequencing were performed.The strains included 27 pathogenic strains(24.5％)and 83 non-pathogenic strains(75.5％).Non-pathogenic strains accounted for a high proportion,particularly among strains from patients(15/16,93.8％).The biological serotypes of pathogenic strains were mainly type 3/O∶3(26/27,96.3％).Non-pathogenic strains included 1A/O∶8 type(23/83,27.7％),1 A/O∶5 type(14/83,16.9％),and the other four biological serotypes(excluding unknown se-rotypes).Pathogenic strains were dominated by type 3/O∶3(26/27,96.3％),and more than 80％of these strains were sensi-tive to 19 antibiotic types.Whole-genome sequencing indicated that the pathogenic strains were all ST135 type,whereas the non-pathogenic strains were more diverse and scattered.HierCC clustering analysis grouped all strains into three clus-ters:pathogenic strains were in one cluster,and strains from patients were found in all three clusters.In conclusion,the Y.enterocolica strains from patients were primarily non-patho-genic.Non-pathogenic strains showed richer epigenetic and ge-netic diversity than pathogenic strains.The monitoring of these strains should be strengthened to decrease the risk of human infection.