Rapid and ultrasensitive detection of pathogen-associated biomarkers is vital for the early diagnosis and therapy of bacterial infections. Herein, we developed a close-packed and ordered Au@AgPt array coupled with a cascade triggering strategy for surface-enhanced Raman scattering (SERS) and colorimetric identification of the Staphylococcus aureus biomarker micrococcal nuclease (MNase) in serum samples. The trimetallic Au@AgPt nanozymes can catalyze the oxidation of 3,3',5,5'-tetramethylbenzidine (TMB) molecules to SERS-enhanced oxidized TMB (oxTMB), accompanied by the color change from colorless to blue. In the presence of S. aureus, the secreted MNase preferentially cut the nucleobase AT-rich regions of DNA sequences on magnetic beads (MBs) to release alkaline phosphatase (ALP), which subsequently mediated the oxTMB reduction for inducing the colorimetric/SERS signal fade away. Using this "on-to-off" triggering strategy, the target S. aureus can be recorded in a wide linear range with a limit of detection of 38 CFU/mL in the colorimetric mode and 6 CFU/mL in the SERS mode. Meanwhile, the MNase-mediated strategy characterized by high specificity and sensitivity successfully discriminated between patients with sepsis (n = 7) and healthy participants (n = 3), as well as monitored the prognostic progression of the disease (n = 2). Overall, benefiting from highly active and dense "hot spot" substrate, MNase-mediated cascade response strategy, and colorimetric/SERS dual-signal output, this methodology will offer a promising avenue for the early diagnosis of S. aureus infection.

Objective:Comprehensive characterization of B-cell phenotypes and spatial distribution in oral lichen planus (OLP) and related oral lichenoid lesions (OLL)(OLP/OLL), with an emphasis on transcriptomic profiling and functional analysis, to uncover the epigenetic mechanisms underlying B cell-mediated immune regulation within the oral mucosal microenvironment.Methods:Single-cell RNA sequencing raw data were sourced from the GSE211630 database, encompassing samples from 2 cases of erosive OLP (EOLP), 3 cases of non-erosive OLP (NEOLP) and 1 healthy control (NORMAL). Following stringent quality control, the data underwent normalization, selection of highly variable genes and batch effect correction. Subsequent analyses included dimensionality reduction and unsupervised clustering to identify distinct cell populations. This study collected pathological specimens from 3 OLP/OLL patients and 3 healthy controls who were treated at the Department of Oral Medicine, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine from January 2021 to December 2023. Using 10X Genomics Visium HD spatial transcriptomics technology, tissue sections were processed through dewaxing, staining and histological imaging, enabling the reconstruction of nucleic acid structures and the capture of gene expression profiles. Data analysis included quality assessment, gene quantification, normalization, dimensionality reduction and clustering. Furthermore, cell type deconvolution was performed using the robust cell type decomposition algorithm, integrating single-cell transcriptomic data to accurately predict and spatially resolve cell type distributions within the tissue microenvironment.Results:After integrating single-cell data from EOLP, NEOLP and NORMAL, cells were classified into seven major categories: B/plasma cells, endothelial cells, epithelial cells, fibroblasts, myeloid cells, smooth muscle cells and T/natural killer cells. The proportion of B/plasma cells varied significantly among the three groups, accounting for 10.7% (1 693/15 815), 3.8% (833/21 653) and 0.4% (47/11 556) of the total cells respectively. Further clustering analysis of B/plasma cells identified four distinct subpopulations: naive B cells, activated B cells, memory B cells and plasma cells. In the EOLP group, these subpopulations constituted 25.9% (348/1 344), 45.9% (617/1 344), 3.3% (45/1 344) and 24.9% (334/1 344) of the B/plasma cells respectively. In the NEOLP group, they represented 31.6% (195/617), 59.6% (368/617), 0.2% (1/617) and 8.6% (53/617). Howerer, only plasma cells were detected in the NORMAL group. Spatial analysis revealed that B cells were actively involved in the formation of tertiary lymphoid structures (TLS) at various stages in OLP/OLL samples, with a prominent structural organization observed in secondary follicle-like TLS. Within these structures, the expressions of T cells marker gene CD3E and B cells marker gene MS4A1 were significantly elevated. Additionally, in secondary follicle-like TLS, the gene encoding follicular dendritic cell secreted protein, germinal center marker gene B cell lymphoma 6 and the gene for activation induced cytidine deaminase also showed strong expression. In OLP/OLL samples, plasma cell marker gene CD38, immunoglobulin (IGH) G3, IGHG1, IGHM, IGHD, IGHE, imunoglobulin Kappa constant, immunoglobulin alpha 1, immunoglobulin Lambda constant 1 and complement gene C3 all exhibited high levels of expression.Conclusions:Compared to normal mucosa, extensive B-cell infiltration is observed in both OLP and OLL, accompanied by significant differences in B-cell phenotypes and proportions. B cells appear to play a central role in local immune responses, primarily through the formation of TLS. However, the precise functional mechanisms underlying their involvement require further investigation.

Rapid and ultrasensitive detection of pathogen-associated biomarkers is vital for the early diagnosis and therapy of bacterial infections.Herein,we developed a close-packed and ordered Au@AgPt array coupled with a cascade triggering strategy for surface-enhanced Raman scattering(SERS)and colorimetric identification of the Staphylococcus aureus biomarker micrococcal nuclease(MNase)in serum samples.The trimetallic Au@AgPt nanozymes can catalyze the oxidation of 3,3',5,5'-tetramethylbenzidine(TMB)molecules to SERS-enhanced oxidized TMB(oxTMB),accompanied by the color change from colorless to blue.In the presence of S.aureus,the secreted MNase preferentially cut the nucleobase AT-rich regions of DNA sequences on magnetic beads(MBs)to release alkaline phosphatase(ALP),which subsequently mediated the oxTMB reduction for inducing the colorimetric/SERS signal fade away.Using this"on-to-off"triggering strategy,the target S.aureus can be recorded in a wide linear range with a limit of detection of 38 CFU/mL in the colorimetric mode and 6 CFU/mL in the SERS mode.Meanwhile,the MNase-mediated strategy characterized by high specificity and sensitivity successfully discriminated between patients with sepsis(n=7)and healthy participants(n=3),as well as monitored the prog-nostic progression of the disease(n=2).Overall,benefiting from highly active and dense"hot spot"substrate,MNase-mediated cascade response strategy,and colorimetric/SERS dual-signal output,this methodology will offer a promising avenue for the early diagnosis of S.aureus infection.

Objective:Comprehensive characterization of B-cell phenotypes and spatial distribution in oral lichen planus (OLP) and related oral lichenoid lesions (OLL)(OLP/OLL), with an emphasis on transcriptomic profiling and functional analysis, to uncover the epigenetic mechanisms underlying B cell-mediated immune regulation within the oral mucosal microenvironment.Methods:Single-cell RNA sequencing raw data were sourced from the GSE211630 database, encompassing samples from 2 cases of erosive OLP (EOLP), 3 cases of non-erosive OLP (NEOLP) and 1 healthy control (NORMAL). Following stringent quality control, the data underwent normalization, selection of highly variable genes and batch effect correction. Subsequent analyses included dimensionality reduction and unsupervised clustering to identify distinct cell populations. This study collected pathological specimens from 3 OLP/OLL patients and 3 healthy controls who were treated at the Department of Oral Medicine, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine from January 2021 to December 2023. Using 10X Genomics Visium HD spatial transcriptomics technology, tissue sections were processed through dewaxing, staining and histological imaging, enabling the reconstruction of nucleic acid structures and the capture of gene expression profiles. Data analysis included quality assessment, gene quantification, normalization, dimensionality reduction and clustering. Furthermore, cell type deconvolution was performed using the robust cell type decomposition algorithm, integrating single-cell transcriptomic data to accurately predict and spatially resolve cell type distributions within the tissue microenvironment.Results:After integrating single-cell data from EOLP, NEOLP and NORMAL, cells were classified into seven major categories: B/plasma cells, endothelial cells, epithelial cells, fibroblasts, myeloid cells, smooth muscle cells and T/natural killer cells. The proportion of B/plasma cells varied significantly among the three groups, accounting for 10.7% (1 693/15 815), 3.8% (833/21 653) and 0.4% (47/11 556) of the total cells respectively. Further clustering analysis of B/plasma cells identified four distinct subpopulations: naive B cells, activated B cells, memory B cells and plasma cells. In the EOLP group, these subpopulations constituted 25.9% (348/1 344), 45.9% (617/1 344), 3.3% (45/1 344) and 24.9% (334/1 344) of the B/plasma cells respectively. In the NEOLP group, they represented 31.6% (195/617), 59.6% (368/617), 0.2% (1/617) and 8.6% (53/617). Howerer, only plasma cells were detected in the NORMAL group. Spatial analysis revealed that B cells were actively involved in the formation of tertiary lymphoid structures (TLS) at various stages in OLP/OLL samples, with a prominent structural organization observed in secondary follicle-like TLS. Within these structures, the expressions of T cells marker gene CD3E and B cells marker gene MS4A1 were significantly elevated. Additionally, in secondary follicle-like TLS, the gene encoding follicular dendritic cell secreted protein, germinal center marker gene B cell lymphoma 6 and the gene for activation induced cytidine deaminase also showed strong expression. In OLP/OLL samples, plasma cell marker gene CD38, immunoglobulin (IGH) G3, IGHG1, IGHM, IGHD, IGHE, imunoglobulin Kappa constant, immunoglobulin alpha 1, immunoglobulin Lambda constant 1 and complement gene C3 all exhibited high levels of expression.Conclusions:Compared to normal mucosa, extensive B-cell infiltration is observed in both OLP and OLL, accompanied by significant differences in B-cell phenotypes and proportions. B cells appear to play a central role in local immune responses, primarily through the formation of TLS. However, the precise functional mechanisms underlying their involvement require further investigation.

Objective Clarify the basis of the commonly used Tibetan medicinal material"YeGexing",the chemical structure and pharmacological activity were investigated,then provide a basis for standardizing clinical medication,quality control,and rational use of resources.Methods Using literature research;plant taxonomy identification summary of chemical composition investigation and pharmacological activity identification,combined with resource distribution,clinical use status investigation and analysis.Results Tibetan medicine"Yegexing"involved 7 species in 2 families,4 genera,that is Sambucus Linn.from Caprifoliaceae,Senecio L.,Synotis(C.B.Clarke)C.Jeffrey et Y.L.Chen,Saussurea DC.from Compositae.The earliest used"Yegexinggabao"or"white"should be Senecio dianthus.and Senecio solidagineus.in the literature;"Yegexingnabao"or"black"should be Saussurea epilobioides.and Sambucus adnate.;S.raphanifolius.(S.diversifolius.),S.chrysanthemoides.(S.laetus.).S.chinensis.are the main substitutes used in Yunnan,Gansu,and western Sichuan,and are commonly used in the market.YeGexing mainly contains terpenes,flavonoids,alkaloids,phenolic acids and other chemical components;YeGexing black is mainly used for"healing",white is mainly used for"anti-inflammatory",which corresponds to modern pharmacological research on anti-inflammatory,antioxidant,antibacterial and other activities.Conclution In view of the fact that the origin of"Yegexing"involves a variety of plants from different families and genera,"Yegexing"has become a collective name for these plant medicinal materials.According to the lextual results and the research progress on chemical structure and pharmacological activity,from the perspective of conducive to standardizing clinical medication,ensuring efficacy and quality of medicinal materials,its name and variety should be standardized as:"??????(???????????????????/)Yegexinggabao"(that is,the white one),the source is S.dianthus.(S.erythropappa.),S.solidagineus.(S.solidaginea.),S.raphalanifolius.(S.diversifolius.),S.chrysanthemoides.(S.laetus.);"(???????????????????????/)Yegexingnabao"(that is,the black one),the source is S.epilobioides.and S.adnata.and S.chinensis are independent medicines.We should strengthen the investigation of the resources and use status of substitutes in various places,the comparative research on the medicinal material basis and biological activity of different resource species,and standardize their varieties-names-bases to make rational use of their resources.

OBJECTIVE To evaluate the effictiveness and safety of recombinant human endostatin combined with afatinib and teggio in the treatment of advanced lung squamous cell carcinoma. METHODS A total of 25 patients with driver-negative advanced lung squamous cell carcinoma were included in this single-arm prospective study, and the enrolled patients were treated with recombinant human endostatin combined with afatinib and teggio as scheduled. Progression-free survival(PFS), overall survival(OS), disease control rate(DCR), objective response rate(ORR), and adverse reactions(AR) were observed and analyzed. RESULTS The 25 enrolled patients received at least 2 cycles of second-line treatment, and were followed up as of March 31, 2023. Among them, 4 patients had partial remission, 17 patients had stable disease, and 4 patients experienced progressive disease. The ORR confirmed by the researchers was 16%(95%CI, 4.5%−36.1%), DCR was 84%(95%CI, 63.9%−95.5%), and median PFS was 5.3 months(95%CI, 3.5−6.9 months). The median OS had not yet been achieved. The entire group of patients had good treatment tolerance, and the most common level Ⅲ or Ⅳ adverse events related to treatment were leukopenia(20%) and rash(12%), with no reported treatment-related deaths. CONCLUSION Recombinant human endostatin combined with afatinib and teggio in the second line treatment of advanced lung squamous cell carcinoma can prolong the progression free survival period of patients and is relatively safe, which is worth further exploration and promotion.

Objective : To investigate the effect of Qiyu Sanlong Decoction ( QYSLD) on inhibiting lung cancer by regulating tumor immune microenvironment through PD-1 signaling pathway and on Th1 immune response in lung cancer bearing mice． Methods : Lewis lung cancer cell line ( LLC) was used to model subcutaneously implanted tumor in mice．72 mice were randomly divided into Control group ( n = 18 ) ，cisplatin group ( n = 18 ) ，QYSLD group (n = 18) and combined group (n = 18) ．The general survival of mice was observedand recorded ; the specific Th1 cell response to tumor antigen was detected by ELISA method ; the proportion of PD-1 positive cells and the ap- optosis of memory CD4 + T cells in spleen T cells of mice were detected by flow cytometry ; and the number and vol- ume of pulmonary metastatic nodules were counted under an anatomical microscope after 21 days of continuous treatment. Results : QYSLD could improve the general survival of mice bearing lung cancer，up-regulate the im- mune response of CD4 + T cells and the specific killing effect of CD8 + T cells．The proportion of apoptotic cells in PD-1 positive cells and CD4 + memory T cells in spleen T cells of mice was down-regulated，and lung metastasis of mouse tumor cells was inhibited． Conclusion QYSLD can improve the survival status of mice bearing lung cancer and enhance Th1 immune response by mediating PD-1 signaling pathway，improve immune function and inhibit im- mune escape of tumor cells，thus inhibiting the metastasis of lung cancer.

Objective:To investigate an association between glycosylated hemoglobin(HbA1c)level and non-alcoholic fatty liver(NAFL)in the elderly.Methods:In this retrospective case-control study, 5 186 elderly individuals aged 65 years and over meeting the inclusion conditions via health physical examination were successively selected from January to December 2018.They were divided into NAFL group(n=1 731)and non-NAFL group(n=3 455). Waist circumference, body mass index, smoking history, diastolic blood pressure, glomerular filtration rate, serum levels of triglyceride, low density lipoprotein cholesterol, alanine aminotransferase, aspartic aminotransferase, fasting blood glucose and HbA1c were compared between the two groups, and their correlations with NAFL were analyzed.Results:The prevalence of NAFL was 33.4%(1, 731/5, 186). The values of waistline, body mass index, smoking history, diastolic blood pressure, triglyceride, total cholesterol, low density lipoprotein cholesterol, glomerular filtration rate, alanine aminotransferase, aspartate aminotransferase, fasting glucose and HbA1c were higher in the NAFL group than in non-NAFL group(all P<0.05). While levels of creatinine, urea nitrogen and age were lower in the NAFL group than in non-NAFL group( P<0.05). According to the quartile of HbA1c level, these subjects were divided into Q1 to Q4 groups(HbA1c<5.7%, 5.7≤HbA1c<6.0%, 6.0%≤HbA1c<6.5%, HbA1c≥6.5%), and the prevalence of NAFL in the Q1 to Q4 were 22.8%(225/1 120), 27.9%(398/1 429), 36.5%(514/1 409), 45.9%(564/1 228)respectively.The prevalence of NAFL was increased along with the increase in the level of HbA1c( P<0.01). Multivariate Logistic regression analysis showed that after adjusting for age, gender and metabolic components, the risk for developing NAFL was gradually increased in Q2 group, Q3 group, Q4 group versus Q1 group as the following OR value: OR=1.274, 95% CI: 1.004-1.616; OR=1.639, 95% CI: 1.294-2.077; OR=1.787, 95% CI: 1.337-2.389, respectively, all P<0.01. Conclusions:The prevalence of NAFL is positively associated with HbA1c levels in the elderly and HbA1c is an independent risk factor for NAFL disease.

Total 732 subjects aged 30-60 years undergoing health check-up at Beijing Hospital Medical Examination Center in 2009,who had no history of non-alcoholic fatty liver disease (NAFLD) were recruited in the study.According to the quartile of hemoglobin (HGB) level,the subjects were divided into 4 groups:Q1:HGB ≤ 131 g/L (n =192),Q2:HGB ＞ 131 g/L and ≤ 140 g/L (n =178),Q3:HGB ＞ 140 g/L and ≤152 g/L (n =184),Q4:HGB ＞ 152 g/L (n =178).All participants were followed up for 4 years,the prevalence rates of NAFLD in groups Q1,Q2,Q3 and Q4 were 8.3％ (16/192),17.4％ (31/178),23.4％ (43/184) and 25.3％ (45/178),respectively (P ＜0.05).Logistic regression showed that the rates of NAFLD in groups Q2,Q3 and Q4 were 2.32 (1.22-4.41),3.36 (1.81-6.21) and 3.72(2.02-6.87) times higher as group Q1 (P ＜ 0.05).Multiple logistic regression analysis showed that the hemoglobin level,TG and BMI were the independent risk factors of NAFLD.

Objective To analyze the effect of Chinese diabetes risk score in health checkup of elderly population and to explore the risk factors of abnormal glucose metabolism in the elderly patients.Methods Chinese diabetes risk score(C-DRs)screening,glucose tolerance test(OGTT),blood biochemical parameters and history collection were performed in 1 181 elderly people participating the health checkup.The area under the ROC curve(AUC)was used to evaluate the accuracy of the screening method.The effect of different cumulative C-DRs on screening target population was reflected by the Gordon index.Multivariate logistic regression analysis was used to analyze relevant risk factors for the glucose metabolic abnormalities.Results The AUC of screening for diabetes was 0.749(95%CI:0.715-0.782),and the best cut-point value was 32.5 points.The sensitivity was 86.50%,the specificity was 60.84%,and the Gordon index was 0.47(P=0.000).The AUC of screening for the pre-diabetes was 0.760(95%CI:0.733-0.787),and the best cut-point was 33.5 points.The sensitivity was 70.89%,the specificity was 68.72%,and the Gordon index was 0.40(P=0.000).The AUC of screening for MS was 0.797(95% CI:0.772-0.823),and the best cut-point value was 32.5 points.The sensitivity was 83.62%,the specificity was 64.90%,and the Gordon index was 0.49(P=0.000).The AUC of screening for insulin resistance was 0.609(95%CI:0.645-0.734),and the best cut-point value was 30.5 points.The sensitivity was 81.25%,the specificity was 44.81%,and the Gordon index was 0.26(P=0.000).Multiple logistic regression analysis showed that age over 80 years,abdominal obesity(waist circumference,male ≥ 90 cm,female ≥ 85 cm),hypertension,hypertriglyceridemia,family history of diabetes were risk factors for abnormality of glucose metabolism in the elderly.The odd ratio values of the above were 1.557,1.543,1.495,1.569,1.625,1.715(all P<0.05).Conclusions Chinese diabetes risk score may be used to screen for diabetes,metabolic syndrome and insulin resistance in the elderly population.Old age,abdominal obesity,hypertension,hypertriglyceridemia and family history of diabetes are independent risk factors for abnormal glucose metabolism in the elderly population.