Objective: To elucidate the biological basis of traditional Chinese medicine (TCM) syndromes from the perspective of “same syndrome, different diseases” in patients with chronic hepatitis B (CHB), liver cirrhosis (LC), and hepatocellular carcinoma (HCC), thereby providing a complementary approach for the diagnosis and treatment of chronic liver diseases (CLD). Methods: To investigate the dynamic characteristics of TCM syndromes in CLD, transcriptomic profiling of peripheral blood mononuclear cells (PBMCs) was performed from patients with CHB, LC, or HCC presenting with three TCM syndromes: liver gallbladder dampness heat syndrome (LGDHS), liver depression spleen deficiency syndrome (LDSDS), and liver kidney Yin deficiency syndrome (LKYDS). These participants were recruited at Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine between August 1, 2018 and December 31, 2021. Differentially expressed genes (DEGs) were identified using the random variance model (RVM) F test with false discovery rate (FDR) correction. Principal component analysis (PCA) and unsupervised hierarchical clustering were applied to visualize sample grouping. Dynamic network biomarkers (DNB) analysis was employed to detect critical transition stages during syndrome evolution, followed by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses to characterize the functional roles and pathway involvement of the DNB members. Random forest (RF) analysis and the area under the receiver operating characteristic (ROC) curves (AUC) were used to rank the importance of candidate genes. External validation was performed using microarray data from an independent CLD cohort (GSE89377) and RNA-seq data from The Cancer Genome Atlas Liver Hepatocellular Carcinoma (TCGA-LIHC) dataset. Additionally, reverse transcription quantitative polymerase chain reaction (RT-qPCR) was performed on an independent cohort of LC patients to validate the expression levels of the candidate genes. Results: The study included a total of 132 participants. DNB analysis identified LDSDS stage as a critical tipping point during TCM syndrome evolution across CHB, LC, and HCC. The phosphoinositide 3-kinase/protein kinase B (PI3K-AKT) signaling pathway was consistently enriched in the DNB analysis across all three types of CLD, suggesting its potential involvement in the critical transition of TCM syndromes. Among the 24 core DNB members of the PI3K-AKT pathway, four genes—integrin subunit beta 1 (ITGB1), collagen type IV alpha 1 chain (COL4A1), collagen type IV alpha 2 chain (COL4A2), and DNA damage inducible transcript 3 (DDIT3)—were identified by RF analysis (Gini score > 1) and ROC analysis. ROC analysis demonstrated high discriminative ability for distinguishing LGDHS from LKYDS in CHB patients, with AUC of 0.7891 for ITGB1, 0.7070 for COL4A1, 0.7148 for COL4A2, and 0.8945 for DDIT3. In the independent CLD cohort (GSE89377), all four genes showed significant stepwise upregulation from normal to CHB, LC, and HCC (all P < 0.05). In the TCGA-LIHC dataset, their expression progressively increased with tumor stage. RT-qPCR validation in an independent LC cohort (30 LGDHS vs. 30 LKYDS) confirmed that ITGB1, COL4A2, and DDIT3 were significantly upregulated in LKYDS compared with LGDHS (P = 0.0152, 0.0186, and 0.0247, respectively), whereas COL4A1 showed a non-significant upward trend (P = 0.1201). Conclusion This study introduces a novel approach to understanding the molecular features underlying TCM syndrome evolution in CLD. The PI3K-AKT pathway and four identified genes (ITGB1, COL4A1, COL4A2, and DDIT3) play crucial roles in the transition from excess (LGDHS) to deficiency (LKYDS) via the critical LDSDS stage. These findings offer potential quantitative biomarkers and therapeutic targets for TCM syndrome differentiation and may help arrest syndrome progression in CLD.

Objective To establish mouse models exposed to different doses of neodymium oxide via tracheal instillation,and to investigate the mechanisms underlying brain tissue damage induced by neodymium oxide exposure in mice.Methods Forty-eight male C57/BL6 mice were randomly assigned to four groups:the control group,the low-dose group,the medium-dose group,and the high-dose group.The low-dose,medium-dose,and high-dose groups received 62.5 mg/mL,125 mg/mL,and 250 mg/mL neodymium oxide,respectively,via non-exposed tracheal instillation.The control group received an equivalent volume of saline using the same administration method.After 35 days,the mice were euthanized,and brain tissues were collected.RT-PCR was used to assess the mRNA expression changes of Claudin-5 and Occludin.Western blot analysis was performed to evaluate the expression changes of Claudin-5 and Occludin tight junction proteins,as well as the expression changes of MMP-2 and MMP-9 in the brain tissues.Additionally,the expression of the RhoA/ROCK2 signaling pathway and downstream cofilin protein was examined.Changes in oxidative stress markers,including MDA,T-AOC,and NO,were measured using a kit method.Results The mRNA expression of Claudin-5 was significantly reduced in the middle-dose and high-dose groups compared to the control group(P<0.05).Similarly,the mRNA expression of Occludin was significantly lower in the low-dose,medium-dose,and high-dose groups compared to the control group(P<0.05).Additionally,the protein expression of Claudin-5,MMP-2,and Occludin was significantly decreased in the low-dose,medium-dose,and high-dose groups compared to the control group(P<0.05).The protein expression of MMP-9 and RhoA was also signifi-cantly lower in the medium-dose and high-dose groups compared to the control group(P<0.05).Furthermore,the protein expression of ROCK2 and p-cofilin in the high-dose group was significantly lower than that in the control group(P<0.05).The content of MDA and T-AOC was significantly lower in the medium-dose and high-dose groups compared to the control group(P<0.05),and the content of NO in the high-dose group was significantly lower than that in the control group(P<0.05).Conclusion Exposure to neodymium oxide results in increased permeability of the blood-brain barrier in mice,leading to oxidative stress,inflammatory responses,and activation of the RhoA/ROCK2 signaling pathway.

Objective To observe the value of three-dimensional ultrasound combined with microvascular flow imaging for prenatal diagnosis of fetal intracranial anomalies.Methods Totally 118 fetuses with intracranial anomalies diagnosed through cranial MRI/induced labor specimen dissection who underwent prenatal ultrasound examination were retrospectively enrolled.Two-dimensional,three-dimensional ultrasound and microvascular flow imaging manifestations of fetal intracranial anomalies were observed,and the accuracy of three-dimensional ultrasound combined with microvascular flow imaging for prenatal diagnosis of fetal intracranial anomalies was analyzed.Results The accuracy of three-dimensional ultrasound combined with microvascular flow imaging for prenatal diagnosis of fetal intracranial anomalies was 93.22％(110/118),of isolated and non-isolated intracranial anomalies was 97.47％(77/79)and 84.62％(33/39),respectively.Six fetuses were missed diagnosis of malformations of cortical development(1 fetus of tuberous sclerosis,4 of abnormal morphology of the sulci gyrus and 1 of schizencephaly)and 1 fetus of intracranial softening lesion,while posterior fossa arachnoid cyst in 1 fetus was misdiagnosed as mega cisterna magna.Conclusion The accuracy of three-dimensional ultrasound combined with microvascular flow imaging for prenatal diagnosis of fetal intracranial anomalies was relatively high.

Objective To observe prenatal ultrasonic findings of fetal malformation of cortical development(MCD).Methods Totally 37 singleton fetuses with MCD diagnosed by fetal cranial MRI and/or genetic examination were retrospectively enrolled,and prenatal ultrasonic manifestations were observed.Results Among 37 fetuses,29(29/37,78.38％)were found with abnormal Sylvian fissure,26(26/37,70.27％)with widened/asymmetrical lateral ventricles,22(22/37,59.46％)with abnormal surface sulci,14(14/37,37.84％)with abnormal cavity of septum pellucidum(CSP)/corpus callosum(CC),11(11/37,29.73％)with midline deviation/curvature,11(11/37,29.73％)with widened subarachnoid space,10(10/37,27.03％)with head circumference＜2 standard deviation below mean value,9(9/37,24.32％)with abnormalities in the cerebellar hemisphere/vermis,7(7/37,18.92％)with abnormal echoes in the brain parenchyma,7(7/37,18.92％)with midline cysts and 2(2/37,5.41％)with enlarged ganglion eminence.Conclusion Fetal MCD mainly presented as abnormal Sylvian fissure,widened/asymmetrical lateral ventricles and abnormal surface sulci,which might accompanied by abnormal CSP/CC,midline deviation/curvature,widened subarachnoid space and abnormal head circumference in prenatal ultrasound.

Objective To establish mouse models exposed to different doses of neodymium oxide via tracheal instillation,and to investigate the mechanisms underlying brain tissue damage induced by neodymium oxide exposure in mice.Methods Forty-eight male C57/BL6 mice were randomly assigned to four groups:the control group,the low-dose group,the medium-dose group,and the high-dose group.The low-dose,medium-dose,and high-dose groups received 62.5 mg/mL,125 mg/mL,and 250 mg/mL neodymium oxide,respectively,via non-exposed tracheal instillation.The control group received an equivalent volume of saline using the same administration method.After 35 days,the mice were euthanized,and brain tissues were collected.RT-PCR was used to assess the mRNA expression changes of Claudin-5 and Occludin.Western blot analysis was performed to evaluate the expression changes of Claudin-5 and Occludin tight junction proteins,as well as the expression changes of MMP-2 and MMP-9 in the brain tissues.Additionally,the expression of the RhoA/ROCK2 signaling pathway and downstream cofilin protein was examined.Changes in oxidative stress markers,including MDA,T-AOC,and NO,were measured using a kit method.Results The mRNA expression of Claudin-5 was significantly reduced in the middle-dose and high-dose groups compared to the control group(P<0.05).Similarly,the mRNA expression of Occludin was significantly lower in the low-dose,medium-dose,and high-dose groups compared to the control group(P<0.05).Additionally,the protein expression of Claudin-5,MMP-2,and Occludin was significantly decreased in the low-dose,medium-dose,and high-dose groups compared to the control group(P<0.05).The protein expression of MMP-9 and RhoA was also signifi-cantly lower in the medium-dose and high-dose groups compared to the control group(P<0.05).Furthermore,the protein expression of ROCK2 and p-cofilin in the high-dose group was significantly lower than that in the control group(P<0.05).The content of MDA and T-AOC was significantly lower in the medium-dose and high-dose groups compared to the control group(P<0.05),and the content of NO in the high-dose group was significantly lower than that in the control group(P<0.05).Conclusion Exposure to neodymium oxide results in increased permeability of the blood-brain barrier in mice,leading to oxidative stress,inflammatory responses,and activation of the RhoA/ROCK2 signaling pathway.

Objective To explore the potential central mechanisms of functional electrical stimulation(FES)hand cycling in promot-ing upper limb motor recovery after stroke.Methods A total of 35 stroke patients hospitalized in the Rehabilitation Center of Changzhou De'an Hospital from May,2023 to December,2024 were enrolled.They sequentially completed 10-minute FES hand cycling and 10-minute sham stimulation(simple hand cycling)tasks.The task order was randomized via dice rolling.Functional near-in-frared spectroscopy(fNIRS)was used to monitor real-time cerebral hemodynamic changes during both tasks to calculate the overall functional connectivity(FC)strength and the FC strength within regions of interest.Results The overall FC strength was higher during the FES hand cycling task than during the sham task(t=2.591,P<0.05),as well as FC between the ipsilateral primary motor cortex(iM1)and ipsilateral somatosensory association cortex(iSAC),iM1 and contralateral somatosensory association cortex(cSAC),ipsilateral pre-motor and supple-mentary motor cortex(iPMC)and iSAC,iPMC and cSAC,iPMC and ipsilateral primary somatosensory cortex(iS1),iPMC and iM1,and contralateral pre-motor and supplementary motor cortex(cPMC)and iSAC(PFDR<0.05).Conclusion FES hand cycling can promote the cortical function remodeling between cerebral hemispheres and the af-fected hemisphere,to integrate sensory-motor function.

This study investigates the feasibility of the VP8*protein as a subunit vaccine target for porcine rotavirus A(PoRVA),a major causative agent of diarrhea in piglets.The VP8* genes of PoRVA P[13]and P[23]genotype strains were amplified by RT-PCR.These genes were then liga-ted into the pET-28a(+)vector,yielding recombinant plasmids pET-28a-XJWF1-VP8*-P[23]and pET-28a-ShXYW13-VP8*-P[13].These plasmids were subsequently transformed into BL21(DE3)competent cells.The VP8*protein,induced by IPTG,was purified using affinity chroma-tography,and its expression and purification were verified by SDS-PAGE and Western blot.The purified VP8* protein was used to immunize mice,and serum samples were collected after three immunizations.Cross-neutralization assays were conducted to evaluate the ability of the VP8*protein immune serum to neutralize different genotype strains.The results demonstrated the ex-pression of soluble VP8*protein,with SDS-PAGE and Western blot analyses showing that the purified VP8*protein existed in both monomeric(27 kDa)and homodimeric(54 kDa)forms.ELISA results indicated that high levels of antibodies were produced in mice immunized with VP 8*-P[13]and VP8*-P[23]after three immunizations.Serum cross-neutralization assays revealed that the neutralizing titers of PoRVA VP8*-P[13]and VP8*-P[23]immune sera against homol-ogous genotype strains ranged from 1∶4 800 to 1∶19 200,significantly higher than those against heterologous genotype strains(1∶1 200).This suggests that the VP8*protein of different geno-type strains exhibits both antigenic conservation and distinct variability.The data obtained in this study provide a solid foundation for further exploration of the antigenic structure of the PoRVA VP8* protein and the development of novel subunit vaccines.

Objective To explore the potential central mechanisms of functional electrical stimulation(FES)hand cycling in promot-ing upper limb motor recovery after stroke.Methods A total of 35 stroke patients hospitalized in the Rehabilitation Center of Changzhou De'an Hospital from May,2023 to December,2024 were enrolled.They sequentially completed 10-minute FES hand cycling and 10-minute sham stimulation(simple hand cycling)tasks.The task order was randomized via dice rolling.Functional near-in-frared spectroscopy(fNIRS)was used to monitor real-time cerebral hemodynamic changes during both tasks to calculate the overall functional connectivity(FC)strength and the FC strength within regions of interest.Results The overall FC strength was higher during the FES hand cycling task than during the sham task(t=2.591,P<0.05),as well as FC between the ipsilateral primary motor cortex(iM1)and ipsilateral somatosensory association cortex(iSAC),iM1 and contralateral somatosensory association cortex(cSAC),ipsilateral pre-motor and supple-mentary motor cortex(iPMC)and iSAC,iPMC and cSAC,iPMC and ipsilateral primary somatosensory cortex(iS1),iPMC and iM1,and contralateral pre-motor and supplementary motor cortex(cPMC)and iSAC(PFDR<0.05).Conclusion FES hand cycling can promote the cortical function remodeling between cerebral hemispheres and the af-fected hemisphere,to integrate sensory-motor function.

Objective To analyze clinical characteristics and therapeutic strategies for patients with Trichosporon asahii(T.asahii)bloodstream infection(BSI),and provide reference for clinical diagnosis and treatment for such disease.Methods Diagnosis and treatment process of a patient with T.asahii BSI were reported.China National Knowledge Infrastructure(CNKI),Wanfang,PubMed,and Web of Science databases were retrieved using key-words:"Trichosporon asahii"and"bloodstream infection".Patients'age,gender,underlying diseases,immune status,treatment,and clinical outcome as well as antimicrobial susceptibility testing results of T.asahii were ana-lyzed retrospectively.Results The leukemia patient developed T.asahii BSI during chemotherapy,and showed clinical improvement after treatment with fluconazole+flucytosine.A total of 44 cases(43 from literature plus this index case)were analyzed.Among these cases,28 were males,the median age was 54.5 years old.The underlying diseases were predominantly hematological diseases.Catheter,urinary tract,and skin were identified as concurrent culture-positive sites.Azoles exhibited good antimicrobial activity in vitro,with voriconazole showing the strongest activity and associated with significantly higher survival rates.Under the guidance of antimicrobial susceptibility tes-ting results,patients with fluconazole treatment had a higher survival rate.At minimum inhibitory concentrations(MICs)of 4-8 μg/mL of fluconazole,fluconazole combined with other agents was predominantly required.Am-photericin B demonstrated good in vitro activity,but the overall survival rate of patients was low when amphotericin B was used as monotherapy without azoles.Catheters removal,surgical intervention,and neutrophil recovery were influencing factors for enhanced survival rates.Conclusion BSI caused by T.asahii primarily occurs in patients with hematological diseases,with neutropenia being a high-risk factor.Infection sites at catheter,urinary tract,and skin may be infection sources.Clearing the source of infection and promoting the recovery of neutrophils can help to improve survival rates of patients.For voriconazole-intolerant patients,fluconazole monotherapy or in combination with flucytosine/amphotericin B can be used alternatively based on antimicrobial susceptibility testing results.

OBJECTIVE To analyze the risk factors for positive culture of carbapenem-resistant Enterobacteriaceae(CRE)in the intensive care unit(ICU)patients so as to provide scientific technical support for early identification of the patients at high-risk of CRE colonization/infection in clinical practice.METHODS The clinical data and bac-terial culture data were collected from the ICU patients who were hospitalized in Beijing Friendship Hospital from 2021 to 2023 so as to build up retrospective cohorts.The risk factors for positive culture of CRE in the ICU pa-tients were screened out with the use of multivariate logistic regression model,and the predictive effect of the model was evaluated by means of receiver operating characteristic(ROC)curves and Hosmer-Lemeshow test.RESULTS Gastric tube indwelling history(OR=3.992,95％CI:1.894 to 8.121),wards of patients detected with CRE within 7 days during the stay(OR=3.518,95％CI:1.492 to 8.294),renal disease history(OR=2.063,95％CI:1.128 to 3.775),history of isolation of multidrug-resistant organisms(OR=2.400,95％CI:1.240 to 4.647)and use of carbapenem antibiotics(OR=1.141,95％CI:1.088 to 1.196)were the independent risk fac-tors for the positive culture of CRE in the ICU patients.The prediction model that was established based on the risk factors showed favorable discrimination,and the area under the ROC curve(AUC)was 0.773.The actual probability was comparatively consistent with the ideal probability,with the Hosmer-Lemeshow test(x2=9.274,P=0.233).CONCLUSION The model shows remarkable effect on prediction of positive culture of CRE in the ICU patients,and it has high value in early identification of the patients at high risk of CRE colonization/infection in clinical practice.