ObjectiveTo explore the value of a multimodal MRI-based radiomics nomogram for differentiating human epidermal growth factor receptor-2 （HER-2） negative breast cancer molecular subtypes.MethodsA retrospective analysis was conducted on 190 patients with HER-2 negative breast cancer who underwent multimodal MRI examination， and the patients were divided into two molecular subtype groups： a HER-2 low expression group （n=108） and a HER-2 zero expression group （n=82）. The cases were randomly stratified and sampled at a ratio of 7∶3 and divided into a training set of 133 cases and a testing set of 57 cases. The clinical and radiological features of the patients were collected， the radiomics features based on T2-weighted imaging （T2WI）， diffusion-weighted imaging （DWI）， and dynamic contrast-enhanced （DCE）-MRI were extracted， and the clinical-radiological model， unimodal radiomics model， multimodal radiomics model， and combined model were constructed respectively. Then the nomogram combined multimodal radiomics signature （radsocre） with clinical-radiological features was used to construct a visualized predictive model， and the area under the curve （AUC） was used to compare the effectiveness of different models in distinguishing HER-2 low expression and zero expression subtypes.ResultsA significant difference in radscore was demonstrated between the HER-2 low and HER-2 zero expression groups in both the training （P<0.000 1） and testing sets （P<0.01）. The AUC of the multimodal radiomics model in the training set and the testing set were 0.914 and 0.836， respectively， which was superior to any unimodal radiomics model. The nomogram demonstrated great diagnostic efficacy （AUC=0.930 in training set； AUC=0.865 in testing set）.ConclusionA multimodal MRI-based nomogram incorporating radsocre and clinical-radiological features can accurately distinguish the subtypes of HER-2 negative breast cancer.

ObjectiveTo investigate the distribution of traditional Chinese medicine （TCM） syndrome patterns among postmenopausal women with dyslipidemia， and to examine the correlations of lipid parameters with bone mineral density （BMD） and bone metabolism markers across common TCM syndrome patterns， thereby providing a clinical reference for individualized integrative traditional Chinese and western medicine interventions stratified according to TCM syndrome patterns. MethodsA multi-center cross-sectional study was conducted among postmenopausal women aged 45-64 years with dyslipidemia. TCM syndromes were determined using a scale-based scoring method. BMD measurements were collected， and fasting venous blood samples were assayed for lipid parameters including total cholesterol （TC）， triglycerides （TG）， high-density lipoprotein cholesterol （HDL-C）， and low-density lipoprotein cholesterol （LDL-C）， and bone metabolism biomarkers including osteocalcin （OST）， total procollagen type Ⅰ N-terminal propeptide （P1NP）， alkaline phosphatase （ALP）， bone-specific alkaline phosphatase （BALP）， β-isomerized C-terminal telopeptide of type Ⅰ collagen （β-CTX）， 25-hydroxyvitamin D ［25（OH）D］， and thyroid-stimulating hormone （TSH）. Chi-square tests were used to compare the distribution of dyslipidemia across different TCM syndromes. Spearman's rank correlation analysis was performed to evaluate the correlations between TCM syndrome patterns and dyslipidemia， as well as the correlations of lipid parameters with BMD and bone metabolism markers within each syndrome pattern. ResultsA total of 506 postmenopausal women with dyslipidemia were included， comprising 172 cases of spleen-kidney yang deficiency syndrome （33.99%）， 163 cases of phlegm-turbidity obstruction syndrome （32.21%）， 118 cases of liver-kidney yin deficiency syndrome （23.32%）， 33 cases of qi stagnation and blood stasis syndrome （6.52%）， and 20 cases of yin deficiency and yang hyperactivity syndrome （3.95%）. Phlegm-turbidity obstruction syndrome was negatively correlated with TC abnormality （r=-0.120， P=0.007） and positively correlated with TG abnormality （r=0.246， P＜0.001）. Spleen-kidney yang deficiency syndrome and liver-kidney yin deficiency syndrome were both negatively correlated with TG abnormality （r=-0.101， P=0.023； r=-0.116， P=0.009， respectively）. Further correlation analysis confirmed that phlegm-turbidity obstruction syndrome was negatively correlated with TC abnormality （r=-0.120） and significantly positively correlated with TG abnormality （r=0.246）. Both the spleen-kidney yang deficiency syndrome and liver-kidney yin deficiency syndrome were negatively correlated with TG abnormality （r=-0.101 and r=-0.116， respectively）. In patients with phlegm-turbidity obstruction syndrome， HDL-C was negatively correlated with BMD at the left femoral neck （r=-0.178， P=0.023） and at left total hip （r=-0.190， P=0.015）. In patients with liver-kidney yin deficiency syndrome， TG was positively correlated with BMD at the lumbar spine （L1-L4） and left total hip （r=0.214， P=0.020； r=0.221， P=0.016， respectively）. More significant correlations were observed in patients with spleen-kidney yang deficiency syndrome， TG was positively correlated with ALP （r=0.210， P=0.006）； TC was positively correlated with TSH and 25（OH）D （r=0.153， P=0.045； r= 0.231， P=0.002， respectively）； HDL-C was positively correlated with OST and 25（OH）D （r=0.156， P=0.040； r=0.228， P=0.003， respectively）； and LDL-C was positively correlated with TSH and 25（OH）D （r=0.156， P= 0.041； r=0.182， P=0.017， respectively）. In patients with liver-kidney yin deficiency syndrome， HDL-C was negatively correlated with TSH （r=-0.204， P=0.027）. ConclusionThe association between dyslipidemia and skeletal health in postmenopausal women varies across TCM syndromes. Lipid abnormalities in women with phlegm-turbidity obstruction syndrome or liver-kidney yin deficiency syndrome are accompanied primarily by site-specific alterations in BMD， whereas those in women with spleen-kidney yang deficiency syndrome are reflected mainly in changes in bone metabolism markers. These findings suggest that targeted intervention strategies should be developed according to TCM syndrome characteristics for postmenopausal women with dyslipidemia.

ObjectiveTo investigate the distribution of traditional Chinese medicine （TCM） syndrome patterns among postmenopausal women with dyslipidemia， and to examine the correlations of lipid parameters with bone mineral density （BMD） and bone metabolism markers across common TCM syndrome patterns， thereby providing a clinical reference for individualized integrative traditional Chinese and western medicine interventions stratified according to TCM syndrome patterns. MethodsA multi-center cross-sectional study was conducted among postmenopausal women aged 45-64 years with dyslipidemia. TCM syndromes were determined using a scale-based scoring method. BMD measurements were collected， and fasting venous blood samples were assayed for lipid parameters including total cholesterol （TC）， triglycerides （TG）， high-density lipoprotein cholesterol （HDL-C）， and low-density lipoprotein cholesterol （LDL-C）， and bone metabolism biomarkers including osteocalcin （OST）， total procollagen type Ⅰ N-terminal propeptide （P1NP）， alkaline phosphatase （ALP）， bone-specific alkaline phosphatase （BALP）， β-isomerized C-terminal telopeptide of type Ⅰ collagen （β-CTX）， 25-hydroxyvitamin D ［25（OH）D］， and thyroid-stimulating hormone （TSH）. Chi-square tests were used to compare the distribution of dyslipidemia across different TCM syndromes. Spearman's rank correlation analysis was performed to evaluate the correlations between TCM syndrome patterns and dyslipidemia， as well as the correlations of lipid parameters with BMD and bone metabolism markers within each syndrome pattern. ResultsA total of 506 postmenopausal women with dyslipidemia were included， comprising 172 cases of spleen-kidney yang deficiency syndrome （33.99%）， 163 cases of phlegm-turbidity obstruction syndrome （32.21%）， 118 cases of liver-kidney yin deficiency syndrome （23.32%）， 33 cases of qi stagnation and blood stasis syndrome （6.52%）， and 20 cases of yin deficiency and yang hyperactivity syndrome （3.95%）. Phlegm-turbidity obstruction syndrome was negatively correlated with TC abnormality （r=-0.120， P=0.007） and positively correlated with TG abnormality （r=0.246， P＜0.001）. Spleen-kidney yang deficiency syndrome and liver-kidney yin deficiency syndrome were both negatively correlated with TG abnormality （r=-0.101， P=0.023； r=-0.116， P=0.009， respectively）. Further correlation analysis confirmed that phlegm-turbidity obstruction syndrome was negatively correlated with TC abnormality （r=-0.120） and significantly positively correlated with TG abnormality （r=0.246）. Both the spleen-kidney yang deficiency syndrome and liver-kidney yin deficiency syndrome were negatively correlated with TG abnormality （r=-0.101 and r=-0.116， respectively）. In patients with phlegm-turbidity obstruction syndrome， HDL-C was negatively correlated with BMD at the left femoral neck （r=-0.178， P=0.023） and at left total hip （r=-0.190， P=0.015）. In patients with liver-kidney yin deficiency syndrome， TG was positively correlated with BMD at the lumbar spine （L1-L4） and left total hip （r=0.214， P=0.020； r=0.221， P=0.016， respectively）. More significant correlations were observed in patients with spleen-kidney yang deficiency syndrome， TG was positively correlated with ALP （r=0.210， P=0.006）； TC was positively correlated with TSH and 25（OH）D （r=0.153， P=0.045； r= 0.231， P=0.002， respectively）； HDL-C was positively correlated with OST and 25（OH）D （r=0.156， P=0.040； r=0.228， P=0.003， respectively）； and LDL-C was positively correlated with TSH and 25（OH）D （r=0.156， P= 0.041； r=0.182， P=0.017， respectively）. In patients with liver-kidney yin deficiency syndrome， HDL-C was negatively correlated with TSH （r=-0.204， P=0.027）. ConclusionThe association between dyslipidemia and skeletal health in postmenopausal women varies across TCM syndromes. Lipid abnormalities in women with phlegm-turbidity obstruction syndrome or liver-kidney yin deficiency syndrome are accompanied primarily by site-specific alterations in BMD， whereas those in women with spleen-kidney yang deficiency syndrome are reflected mainly in changes in bone metabolism markers. These findings suggest that targeted intervention strategies should be developed according to TCM syndrome characteristics for postmenopausal women with dyslipidemia.

Objective: To explore the value of nomogram based on DCE-MRI radiomics combined with clinical-ra- diological features in predicting HR status in breast cancer with Her-2 low expression． Methods: A total of 198 pa- tients of Her-2 low expression breast cancer who underwent standardized breast MRI in our hospital from January 2019 to February 2025 were retrospectively analyzed．Patients were divided into HR ( + ) group ( n = 137) and HR ( －) group ( n = 61) ．The cases were divided into a training set ( 138 cases) and a testing set ( 60 cases) in a 7 ∶ 3 ratio．Clinical-radiological model was based on clinical and traditional radiological features，radiomics model was based on DCE-MRI，and combined model was constructed，respectively．The nomogram was drawn，and re- ceiver operating characteristic curve was used to compare the performance of different models in predicting HR sta- tus． Results: The DCE-MRI radiomics score ( Radscore) between the HR ( + ) group and the HR ( －) group showed statistical differences in both the training and testing sets ( both P＜0. 001) ．The AUC of the clinical-radio- logical model based on lesion mobility，Ki67，TIC type，enhancement pattern and maximum diameter for predicting HR status in the training set and testing set were 0. 643 and 0. 616，respectively．The AUC of the DEC-MRI ra- diomics model in the training set and testing set were 0. 897 and 0. 860，respectively．The nomogram drawn by combining clinical-radiological features and Radscore showed better predictive performance in both the training set ( AUC = 0. 913) and testing set ( AUC = 0. 898) than the clinical-radiological model ( all P＜0. 05) ． Conclusion The nomogram combined by DCE-MRI radiomics and clinical-radiological features can effectively predict HR sta-tus of breast cancer with low Her-2 expression，which is helpful to the building of individualized treatment plan for breast cancer patients．

Objective:To investigate the effects of decellularised extracellular matrix material(dECM)extract on macrophage survival,migration,phagocytosis,pro-inflammatory factor expression and ROS production.Methods:CCK-8 method was used to detect whether the extract of dECM material had cytotoxicity on macrophages;effects of dECM material extracts on macrophage recruit-ment and chemotaxis were examined by Transwell migration assay;effects of dECM material extract on macrophage phagocytosis were detected by pHrodo staining;effects of dECM material extract on expression of pro-inflammatory genes and pro-inflammatory specific functional molecules were detected by RT-qPCR and immunofluorescence staining;detection of the effect of dECM material extracts on macrophage ROS production by the DCFH-DA fluorescent probe approach.Results:Compared with DMEM complete medium,①CCK-8 assay showed that dECM material extract had no toxic effect on macrophages,and could promote the formation of macrophage colonies;②pHrodo staining assay showed that dECM material extract did not affect phagocytosis of macrophages;③Transwell migra-tion assay showed that dECM material extract did not promote macrophage migration;④RT-qPCR results showed that dECM material extracts were able to down-regulate gene expressions of pro-inflammatory cytokines and specific functional molecules;⑤Flow cytome-try results showed that dECM material extract could reduce the production of ROS by macrophages.dECM material extract had excel-lent biocompatibility for macrophages.Conclusion:dECM material extract is non-toxic to macrophages and is able to reduce macro-phage pro-inflammatory gene expression and ROS production.

Objective:To investigate the effects of decellularised extracellular matrix material(dECM)extract on macrophage survival,migration,phagocytosis,pro-inflammatory factor expression and ROS production.Methods:CCK-8 method was used to detect whether the extract of dECM material had cytotoxicity on macrophages;effects of dECM material extracts on macrophage recruit-ment and chemotaxis were examined by Transwell migration assay;effects of dECM material extract on macrophage phagocytosis were detected by pHrodo staining;effects of dECM material extract on expression of pro-inflammatory genes and pro-inflammatory specific functional molecules were detected by RT-qPCR and immunofluorescence staining;detection of the effect of dECM material extracts on macrophage ROS production by the DCFH-DA fluorescent probe approach.Results:Compared with DMEM complete medium,①CCK-8 assay showed that dECM material extract had no toxic effect on macrophages,and could promote the formation of macrophage colonies;②pHrodo staining assay showed that dECM material extract did not affect phagocytosis of macrophages;③Transwell migra-tion assay showed that dECM material extract did not promote macrophage migration;④RT-qPCR results showed that dECM material extracts were able to down-regulate gene expressions of pro-inflammatory cytokines and specific functional molecules;⑤Flow cytome-try results showed that dECM material extract could reduce the production of ROS by macrophages.dECM material extract had excel-lent biocompatibility for macrophages.Conclusion:dECM material extract is non-toxic to macrophages and is able to reduce macro-phage pro-inflammatory gene expression and ROS production.

Objective To study the immunoadjuvant effects of chitosan oligosaccharide(COS),including the immune activation and the triggering of lysosomal escape,and to explore whether COS can be used as an adjuvant for attenuated live bacteria vector vaccines.Methods 1)Mouse macrophages RAW264.7 cells were cultured with COS at 0 mg/mL(the control group)and 0.1-4 mg/mL for 24 h and the effect on cell viability was measured by CCK8 assay.Mouse macrophages RAW264.7 were treated with COS at 0(the control group),1,2,and 4 mg/mL for 24 h.Then,the mRNA expression levels of the cytokines,including IFN-γ,IL-10,TGF-β,and TLR4,were determined by RT-qPCR assay.2)RAW264.7 cells were treated with 1 mL of PBS containing different components,including calcein at 50 μg/mL,COS at 2 mg/mL,and bafilomycin A1,an inhibitor,at 1 μmol/mL,for culturing.The cells were divided into the Calcein group,Calcein+COS group,and Calcein+COS+Bafilomycin A1 group accordingly.Laser scanning confocal microscopy was used to observe the phagocytosis and the intracellular fluorescence distribution of calcein,a fluorescent dye,in RAW264.7 cells in the presence or absence of COS intervention to determine whether COS was able to trigger lysosomal escape.3)LM?E6E7 and LI?E6E7,the attenuated Listeria vector candidate therapeutic vaccines for cervical cancer,were encapsulated with COS at the mass concentrations of 0.5 mg/mL,1 mg/mL,2 mg/mL,4 mg/mL,and 8 mg/mL.Then,the changes in zeta potential were measured to select the concentration of COS that successfully encapsulated the bacteria.Phagocytosis of the vaccine strains by RAW264.7 cells was measured before and after LM?E6E7 and LI?E6E7 were coated with COS at 2 mg/mL.Results 1)CCK8 assays showed that,compared with the findings for the control group,the intervention of RAW264.7 cells with COS at different concentrations for 24 h was not toxic to the cells and promoted cell proliferation,with the difference being statistically significant(P＜0.05).According to the RT-qPCR results,compared with those of the control group,the COS intervention up-regulated the mRNA levels of TLR4 and IFN-γ in RAW264.7 cells,while it inhibited the mRNA expression levels of TGF-β and IL-10,with the most prominent effect being observed in the 4 mg/mL COS group(P＜0.05).2)Laser scanning confocal microscopy revealed that the amount of fluorescent dye released from lysosomes into the cells was greater in the Calcein+COS group than that in the Calcein group.In other words,a greater amount of fluorescent dye was released from lysosomes into the cells under COS intervention.Furthermore,this process could be blocked by bafilomycin A1.3)The zeta potential results showed that COS could successfully encapsulate the surface of bacteria when its mass concentration reached 2 mg/mL.Before and after the vaccine strain was encapsulated by COS,the phagocytosis of LM?E6E7 by RAW264.7 cells was 5.70%and 22.00%,respectively,showing statistically significant differences(P＜0.05);the phagocytosis of LI?E6E7 by RAW264.7 cells was 1.55%and 6.12%,respectively,showing statistically significant differences(P＜0.05).Conclusion COS has the effect of activating the immune response of macrophages and triggering lysosomal escape.The candidates strains of coated live attenuated bacterial vector vaccines can promote the phagocytosis of bacteria by macrophages.Further research is warranted to develop COS into an adjuvant for bacterial vector vaccine.

Objective:To investigate the changes of inflammation and immune function in children with chronic tonsillitis after tonsillotomy. Methods:Prospectively collected 60 children with obstructive sleep apnea （OSA） diagnosed as chronic tonsillitis with adenoids and tonsillar hypertrophy from January to June 2021. Two groups were divided, the experimental group （n=30） underwent bilateral partial tonsillectomy + adenoidectomy by hypothermia plasma ablation, and the control group （n=30） underwent adenoidectomy by using the same hypothermia plasma ablation method. The number of tonsillitis attacks before surgery and within one year after surgery was recorded, and the serum immunoglobulin IgM, IgG, IgA, complement C3 and complement C4 levels before operation, one month and three months after operation were measured. Results:The number of tonsillitis attacks in the experimental group and the control group at one year after surgery was lower than that before surgery（P<0.05）; The number of inflammatory attacks in the experimental group was （0.50±0.63） times/year, which was lower than that of （1.33±0.80） times/year in the control group. There was no significant difference in the five immunization results of the two groups at one month and three months after operation compared with before operation, and there was also no significant difference between the experimental and the control groups. Conclusion:Partial tonsillectomy can be applied to children with chronic tonsillitis, which can effectively reduce the number of tonsillitis attacks and has no effect on the immune function of children.
Child ; Humans ; Tonsillectomy/methods* ; Hypothermia ; Tonsillitis/surgery* ; Adenoidectomy ; Palatine Tonsil/surgery* ; Inflammation ; Chronic Disease ; Immunity

Objective To construct Listeria monocytogenes(LM)and Listeria ivanovii(LI)balanced lethal systems expressing cervical cancer antigens,to study their basic biological characteristics,and to provide reference data for the immunotherapy of cervical cancer.Methods Through seamless cloning via in vitro ligation kit,the HPV16 E6E7 fusion protein antigen gene constructed in our lab was spliced to the complement plasmid pCWgfp-LM dal-Amp that contained the nutritional gene dal.Then,we replaced the ampicillin(Amp)resistance gene of the complement plasmid with the asd nutrition gene.The ligation reaction mixture was transformed into Escherichia coli(E.coli)recipient bacteria DH5аΔasd and the complement plasmid pCWgfp-E6E7-LM dal-Ampfree,which expressed cervical cancer antigens and had no Amp resistance,was obtained by nutrition screening from the E.coli DH5аΔasd.The plasmid pCWgfp-E6E7-LM dal-Ampfree was complemented into LMΔdd and LIΔdd,the attenuated nutrition-deficient Listeria strains with the virulence genes actA and plcB and nutrition genes dal and dat deleted by electroporation,thereby obtaining LM and LI balanced lethal systems expressing cervical cancer antigen genes.The in vitro growth of the strains was observed.Western blot was performed to examine the status of antigen protein expression.PCR was performed to measure the in vitro passage stability of complement plasmid pCWgfp-E6E7-LM dal-Ampfree.Their basic biological characteristics were examined by biochemical reaction tests and hemolysis assay.Results Two Listeria balanced lethal systems expressing cervical cancer antigen were successfully constructed.The HPV16 type E6E7 fusion protein was successfully expressed in the two Listeria balanced lethal systems.pCWgfp-E6E7-LM dal-Ampfree,the positive plasmid expressing cervical cancer antigen,maintained stable existence in the two Listeria balanced lethal systems.The two Listeria balanced lethal systems expressing cervical cancer antigen showed significantly better recovery growth in comparison with Listeria nutrition deficiency strains.The results of biochemical reaction tests showed that most of the biochemical reaction of the two Listeria balanced lethal systems expressing cervical cancer antigen were consistent with those of Listeria attenuated strains.The two Listeria balanced lethal systems expressing cervical cancer antigen still maintained the hemolytic ability,although their hemolytic ability was slightly inferior to that of the Listeria balanced lethal systems not expressing cervical cancer antigen and the Listeria attenuated strains.Conclusion The two Listeria balanced lethal systems expressing cervical cancer antigen genes are constructed successfully.They display normal in vitro growth.The complement plasmid pCWgfp-E6E7-LM dal-Ampfree can maintain stable existence in vitro,showing little change in its biochemical characteristics and hemolytic ability.Further research should be conducted to investigate the potential of these two recombinant strains to be used as candidate strains for cervical cancer therapeutic vaccine.

Objective: To analyze the changes in the expression of hypoxia inducible factor-1α (HIF-1α) and inflammatory cytokines and to investigate the role of HIF-1α in regulating the production of inflammatory cytokines during influenza A (H1N1) virus infection. Methods: BALB/c mice were injected with H1N1 virus to establish the mouse model of H1N1 virus infection. Fifteen BALB/c mice were randomly divided into three groups: control group, H1N1 virus group and H1N1 virus+ HIF-1α inhibitor group. Inflammatory cytokines (IL-6, TNF-α, IL-1β and IL-10) in samples of serum and lung tissues were detected by Luminex and ELISA. Levels of HIF-1α in serum and lung tissue samples were detected by Western blot and ELISA, respectively. Results: Compared with the control group, the levels of inflammatory cytokines in serum (IL-6, TNF-α, IL-1β and IL-10) and lung tissues (IL-6 and TNF-α) and the expression of HIF-1α in serum and lung tissues in the H1N1 virus group were significantly increased. The levels of HIF-1α, IL-6, TNF-α IL-1β and IL-10 in lung tissues in H1N1 virus+ HIF-1α inhibitor group were significantly lower than those of the H1N1 virus group. Conclusion During H1N1 virus infection, the levels of inflammatory cytokines and HIF-1α were significantly increased. The production of inflammatory cytokines was significantly reduced after inhibiting HIF-1α expression, suggesting that HIF-1α might promote the production of inflammatory cytokines.