BACKGROUND: Peripheral helper T (T PH ) cells are uniquely positioned within pathologically inflamed non-lymphoid tissues to stimulate B-cell responses and antibody production. However, the phenotype, function, and clinical relevance of T PH cells in hepatocellular carcinoma (HCC) are currently unknown. METHODS: Blood, tumor, and peritumoral liver tissue samples from 39 HCC patients (Sep 2016-Aug 2017) and 101 HCC patients (Sep 2011-Dec 2012) at the Third Affiliated Hospital of Sun Yat-sen University were used. Flow cytometry was used to quantify the expression, phenotype, and function of T PH cells. Log-rank tests were performed to evaluate disease-free survival and overall survival in samples from 39 patients and 101 patients with HCC. T PH cells, CD19 + B cells, and T follicular helper (T FH ) cells were cultured separately in vitro or isolated from C57/B6L mice in vivo for functional assays. RESULTS: T PH cells highly infiltrated tumor tissues, which was correlated with tumor size, early recurrence, and shorter survival time. The tumor-infiltrated T PH cells showed a unique ICOS hi CXCL13 + IL-21 - MAF + BCL-6 - phenotype and triggered naïve B-cell differentiation into regulatory B cells. Triggering programmed cell death protein 1 (PD-1) induced the production of C-X-C motif chemokine ligand 13 (CXCL13) by T PH cells, which then suppressed tumor-specific immunity and promoted disease progression. CONCLUSION Our study reveals a novel regulatory mechanism of T PH cell-regulatory B-cell-mediated immunosuppression and provides an important perspective for determining the balance between the differentiation of protumorigenic T PH cells and that of antitumorigenic T FH cells in the HCC microenvironment.
Carcinoma, Hepatocellular/metabolism* ; Liver Neoplasms/metabolism* ; Humans ; T-Lymphocytes, Helper-Inducer/metabolism* ; Animals ; Mice ; Male ; Female ; Mice, Inbred C57BL ; Middle Aged ; B-Lymphocytes, Regulatory/metabolism* ; Flow Cytometry ; Interleukin-21 ; Aged ; Chemokine CXCL13/metabolism*

The Hedgehog (Hh) signaling pathway is a critical transduction system regulating cell proliferation, differentiation, and tissue homeostasis during embryonic development. Its aberrant activation is closely associated with the pathogenesis of malignancies such as basal cell carcinoma and medulloblastoma. Although Smoothened (SMO)-targeting inhibitors have received clinical approval, their therapeutic efficacy is limited by acquired resistance mutation and compensatory pathway activation. Glioma-associated oncogene homolog 1 (GLI1), the terminal effector transcription factor of the Hh pathway, has emerged as a promising therapeutic target due to its tumor-specific overexpression and lower propensity for resistance induction. However, GLI1 is classified as an "undruggable" target due to the absence of well-defined ligand-binding pockets, low inhibitory activity, and poor drug-like properties. Currently, no GLI1 inhibitor has entered clinical trials. This review systematically analyzes multidimensional modulation strategies (e.g., allosteric modulation, protein-protein interaction disruption, targeted protein degradation) for targeting transcription factors, based on the structural and functional features of GLI1-DNA interaction combined with recent advances in structural biology and chemical biology, offering new paradigms to overcome therapeutic barriers against undruggable targets.

Urine is produced from the urinary system, and urinary cell-free DNA (cfDNA) carries genomic DNA directly secreted from urinary system.Urine samples are non-invasive, unlimited in quantity and easy to obtain, making urinary cfDNA a promising biomarker for urologic diseases.This article reviews the progress of clinical application of urinary cfDNA in urologic diseases.

Myopia is becoming more and more common all over the world, and the incidence of myopia is gradually increasing. Many treatments have been used to prevent and control myopia, including optics, drugs, environment or behavior, but the results are different and lack standardization. At present, many experiments have proved that peripheral defocus technology has a certain effect on myopia control. Based on this technology, three kinds of framed eyeglass lenses with peripheral defocus design, namely defocus incorporated multiple segments(DIMS), highly aspherical lenslets(HAL)and cylindrical annular reactive elements(CARE), are commonly used in medical and optometry institutions in China. These lenses provide not only clear vision in the central area, but also a certain amount of myopic defocus in the periphery to control the progression of myopia. This paper aims to focus on the design principle and myopia prevention and control effect of the above three peripheral defocus lenses, and evaluate their effectiveness in clinical practice.

Objective·To study the differences in the structure and load of hair follicle microbiota in non-lesional areas among patients with moderate-to-severe acne vulgaris and healthy individuals,and to explore the relationship between microorganisms and the severity of acne vulgaris.Method·A cross-sectional study was used.Patients with moderate or severe acne vulgaris(referred to as acne)and healthy volunteers who visited the Department of Dermatology,Renji Hospital,Shanghai Jiao Tong University School of Medicine,from August 2022 to August 2023.16S rRNA high-throughput sequencing and quantitative real-time polymerase chain reaction(qPCR)were performed on the follicular contents from the non-lesional areas of the faces of patients with moderate or severe acne and healthy volunteers to analyze the diversity,species composition,and microbial load differences in hair follicle bacteria in patients with different severity of acne.Results·Ten patients with moderate acne,eleven patients with severe acne,and eleven healthy volunteers were included.There were no statistically differences in general data such as age and gender ratio among the three groups.Bacterial α-diversity was significantly lower in both the moderate and severe acne groups compared to the healthy group(P=0.020,P=0.013).The principal coordinates analysis(PCoA)plot showed that the sample distribution of the healthy group was relatively concentrated,with small differences within the group,and the distribution of samples in the moderate and severe acne groups exhibited a certain trend but was relatively scattered,with differences between the groups.There were differences in the trend distribution of the three sample groups,and there were differences in the microbial community structure between the groups.The results of similarity analysis showed significant differences in β-diversity and low similarity in species composition between the healthy and moderate acne groups(P=0.027)and between the healthy and severe acne groups(P=0.017),and high species similarity between the moderate acne and severe acne groups(P=0.160).The dominant bacterial groups at the phylum level were Actinobacteria,Firmicutes,Proteobacteria,and Bacteroidetes.At the genus level,the dominant bacteria in the healthy group were Propionibacterium and unclassified Actinomycetales,and the dominant bacteria in both acne groups were Staphylococcus and Propionibacterium.Compared to the healthy group,the relative abundance of Staphylococcus species in the hair follicles in non-lesional areas of the moderate and severe acne groups was significantly increased(P=0.010,P=0.019).Compared with the healthy control group,the hair follicle microbiota load in non-lesional areas of both the moderate and severe acne groups was significantly increased(both P=0.001).Compared with the moderate acne group,the bacterial load in the hair follicle samples of the severe acne group was significantly increased(P=0.017).Conclusion·The microbial community structure of hair follicles in non-lesional areas of patients with moderate or severe acne is different from that of healthy individuals,and the microbial diversity in the acne group is significantly reduced.The relative abundance of Staphylococcus species in the hair follicles in non-lesional areas of the moderate or severe acne groups is significantly increased compared to the healthy group.As the severity of acne increases,the bacterial load in hair follicles in non-lesional areas significantly increases.This research suggests that the occurrence and severity of acne may be related to the community structure and load of hair follicle microbiota.

AIM:To investigate the effects of Panax notoginseng compound formula(PN)on endometrial fibro-sis by regulating inflammatory reaction and intestinal flora(IF)in a rat model of intrauterine adhesion(IUA).METHODS:The rat IUA model was established by following the mechanical injury method.A total of 50 rats were randomly divided in-to sham group,model group,low-dose(210 mg/kg)PN group,medium-dose(420 mg/kg)PN group and high-dose(840 mg/kg)PN group.After 8 weeks of intragastric administration,the uterus was collected to observe morphological changes with naked eye.The degree of uterine tissue damage and fibrosis was evaluated through hematoxylin-eosin(HE)and Mas-son staining.The collagen type Ⅰ(Col Ⅰ)was detected by immunohistochemistry.The interleukin-6(IL-6)and IL-10 pro-tein expression was detected by Western blot.The levels of IL-6,IL-1β,tumor necrosis factor-α(TNF-α)and vascular endothelial growth factor B(VEGFB)were detected by enzyme-linked immunosorbent assay(ELISA).The IF diversity and population structure were observed by 16S amplicon.RESULTS:Compared with the sham group,the uteruses of rats in the model group showed:reduced elasticity,accompanied by congestion and edema;decreased number of glands and blood vessels,and thinned endometrium(P<0.01);increased collagen fibers and Col Ⅰ protein expression(P<0.01);sig-nificantly increased IL-1β,IL-6,TNF-α and VEGFB levels in the uterine tissue(P<0.01);decreased IL-10 level(P<0.01);and reduced IF diversity(P<0.05).Compared with the model group,the drug intervention groups exhibited:re-covered elasticity of the uterus and relieved congestion and edema;increased number of endometrial glands and blood ves-sels(P<0.05);decreased collagen fibers and Col Ⅰ protein expression(P<0.01);reduced IL-1β,IL-6,and TNF-α lev-els to varying degrees in the uterine tissue(P<0.05);elevated IL-10 level(P<0.01);and improved IF diversity(P<0.05).CONCLUSION:The PN is able to significantly improve the endometrial tissue fibrosis in IUA rats.The under-lying mechanisms may be related to the inhibition of IL-6,IL-1β and TNF-α expression,up-regulation of IL-10,and im-provement of IF diversity.

Objective To observe the value of spectral CT multi-parameter imaging for preoperative predicting lymph node metastasis(LNM)of gastric cancer.Methods Totally 136 patients with gastric adenocarcinoma were retrospectively enrolled.The patients were further divided into LNM group(n=74)and non-LNM group(n=62)according to postoperative pathological findings of lymph nodes status.Clinical data,conventional CT findings and spectral CT parameters were compared between groups.Factors being significant different between groups were included in multivariate logistic regression analysis to screen independent predictors of gastric cancer LNM.Clinical+conventional CT model(model 1),spectrum CT model(model 2)and combined model(model 3)were constructed based on the above independent predictors,respectively.Receiver operating characteristic curve was drawn,and the area under the curve(AUC)was calculated to evaluate the efficacy of each model for preoperative predicting LNM of gastric cancer.Results CT-N stage,CT-T stage,70,100 and 140 keV CT valuestumor at arterial phase(AP),arterial enhancement fraction(AEF)and normalized iodine concentration at venous phase(NICVP)were all independent predictors of gastric cancer LNM(all P＜0.05).AUC of model 3 was 0.846,higher than that of model 1 and model 2(AUC=0.767,0.774,Z=-0.368,-2.373,both P＜0.05)for preoperative predicting LNM of gastric cancer,while the latter two were not significantly different(Z=-0.152,P=0.879).Conclusion Spectral CT multi-parameter imaging could effectively predict LNM of gastric cancer preoperatively.

Objectives:To investigate the relationship between paraspinal muscle degeneration and isthmic spondylolisthesis in L5/S1.Methods:The clinical data of 108 patients with L5/S1 bilateral isthmic spondylolysis admitted and treated at the Department of Orthopedics,the First Affiliated Hospital of Zhengzhou University from January 1,2020 to December 31,2023 were retrospectively analyzed.There were 62 females and 46 males,aged 26-86(52.8±10.4)years.All the patients had complete imaging data,including lateral and dynamic lumbar X-ray,lumbar CT and lumbar MRI.According to the presence or absence of spondylolisthesis and Meyerding grading,the patients were divided into non-spondylolisthesis group(28 cases),grade Ⅰ spondylolisthesis group(46 cases)and grade n spondylolisthesis group(34 cases).Venebral body cross-sectional area(V CSA),erector spinae muscle cross-sectional area(ES CSA),multifidus muscle cross-sectional area(MF CSA),and facet joint angle(FJA)were measured at the inferior vertebral endplate of L5 vertebral body on the axial view of lumbar spine MRI.And the mean FJA(mFJA),relative MF CSA(MF rCSA),relative ES CSA(ES rCSA),fat infiltration rate of multifidus muscle(MF FIR)and fat infiltration rate of erector spinae muscle(ES FIR)were calculated.The differences in age,gender,body mass index(BMI),disease duration,V CSA,mFJA,MF CSA,ES CSA,MF rCSA,ES rCSA,MF FIR and ES FIR between the three groups were compared.Ordinal logistics regression was used to analyze the relationship between age,gender,disease duration,BMI,mFJA,MF rCSA,ES rCSA,MF FIR,ES FIR and the isthmic spondylolisthesis.Results:There were no statistical differences in age,body weight,BMI,V CSA,and mFJA between the three groups(P＞0.05).The proportion of female patients in the non-spondylolisthesis group(29％)was significantly lower than that in grade Ⅰ spondylolisthesis group(61％)and grade Ⅱ spondylolisthesis group(76％)(P＜0.05).The height of non-spondylolisthesis group(1.68±0.06m)was higher than that of grade Ⅰ spondylolisthesis group(1.64±0.75m)and grade Ⅱ spondylolisthesis group(1.63±0.54m)(P＜0.05),and the disease duration in the non-spondylolisthesis group was shorter than that of grade Ⅰ spondylolisthesis group and grade Ⅱspondylolisthesis group(P＜0.05).There was no significant difference in MF CSA,MF rCSA and ES FIR between the three groups(P＞0.05).ES CSA and ES rCSA in grade Ⅱ spondylolisthesis group(3183.2±1196.38mm2 and 2.14±1.11)were significantly greater than that in grade Ⅰ spondylolisthesis group(2579.28±896.56mm2 and 1.65±0.65)and non-spondylolisthesis group(2401.40±742.98mm2 and 1.56±0.58)(P＜0.05).The MF FIR of grade Ⅱ spondylolisthesis group(40.70±14.05)％was significantly higher than that of grade Ⅰspondylolisthesis group(30.39±12.98)％and non-spondylolisthesis group(26.69±9.96)％(P＜0.05).There were no significant differences in ES CSA,ES rCSA and MF FIR between grade Ⅰ spondylolisthesis group and non-spondylolisthesis group(P＞0.05).Ordinal logistics regression analysis showed that female(OR=2.81;95％CI:1.120-7.062;P=0.028)and increased MF FIR(OR=1.044;95％CI:1.003-1.088;P=0.034)were the independent risk factors for the progression of isthmic spondylolisthesis(P＜0.05).Conclusions:Patients with a higher degree of isthmic spondylolisthesis have more severe fat infiltration of multifidus muscle and hypertrophy of erector spinae muscle.Female and increased MF FIR are independent risk factors for the progression of isthmic spondylolisthesis.

Objective:To invastigate the targets and signaling pathways of Gubiao Fanggan Modified Formula in regulating the defense against influenza A virus in immunosuppressed mice by network pharmacology,and the key targets were verified by immuno-suppressive mice model.Methods:TCMSP database was used to search the active ingredients of Gubiao Fanggan Modified Formula,and GeneCard,OMIM,PharmGkb databases were used to obtain the potential targets of the active ingredients to prevent influenza,and take their intersection targets were taken;STRING11.5 database was used to make protein-protein interaction network analyzed and finded the core targets;Cytoscape3.8.1 was used to build a traditional Chinese medicine-ingredient-disease target network,and GO enrichment analysis and KEGG enrichment analysis were performed.Intraperitoneal injection of cyclophosphamide was used to construct a mouse model of immune function suppression,normal group,model control group,Gubiao Fanggan Modified Formula group and oseltamivir group were set up,followed by prophylactic administration,and influenza virus intervention was performed on the fourth day.After 7 days of intragastric administration,the key targets were verified by mouse spleen index,HE staining,RT-qPCR and immunohistochemistry.Results:There were 82 active ingredients in five traditional Chinese medicines in Gubiao Fanggan Modi-fied Formula,and 72 common targets of drugs and diseases such as IL-6,TNF-α,IL-2,etc,mainly involving IL-17,TNF and AGE-RAGE signaling pathway.Gubiao Fanggan Modified Formula could increase spleen index and significantly reduce mRNA and protein expressions of IL-6 and TNF-α in spleen tissue of mice(P<0.05 or P<0.01).Conclusion:Gubiao Fanggan Modified Formula may regulate body's immune function through targets such as IL-6 and TNF-α,thereby preventing influenza virus infection.

Objective To observe the value of dual-energy CT(DECT)radiomics combined with clinical and CT features for predicting differentiation degree of gastric adenocarcinoma(GAC).Methods Totally 254 patients with GAC were prospectively analyzed and divided into high-grade group(low differentiation GAC,n=88)and low-grade group(middle-high differentiation GAC,n=166)according to pathological results.The patients were also divided into training set(n=203,including 70 high-grade and 133 low-grade GAC)and verification set(n=51,including 18 high-grade and 33 low-grade GAC)at the ratio of 8∶2.Radiomics features were extracted and screened based on venous phase single-level(40,70,100 and 140 keV)DECT,and a multi-energy radiomics model was constructed to predict GAC classification.Univariate analysis and multivariate logistic regression were used to analyze clinical and CT features as well as DECT parameters in training set to construct a clinic-CT model.Then a combined model was constructed through combining clinic-CT model with radiomics model.The predictive efficacy of the models were evaluated,and the calibration degree of the combined model was assessed.Results The area under the curve(AUC)of clinic-CT model,multi-energy radiomics model and combined model was 0.74,0.75 and 0.78 in training set,and 0.73,0.77 and 0.78 in verification set,respectively.Except for AUC of combined model was higher than that of clinic-CT model in training set(P＜0.05),no significant difference of AUC was found among models in training set nor verification set(all P＞0.05).The calibration degree of combined model was good in both training set and verification set(both P＞0.05).Conclusion DECT radiomics combined with clinical and CT features could effectively predict differentiation degree of GAC.