Objective·To construct a framework nucleic acid-based linear amplification platform for the sensitive and quantitative detection of bladder cancer-related microRNAs(miRNAs),facilitating early screening and accurate diagnosis of bladder cancer.Methods·This study combined a plasma fluorescence-enhanced chip with high-performance tetrahedral framework nucleic acid(tFNA)probes,targeting miRNAs as biomarkers,to construct a framework nucleic acid-based linear signal amplification platform for precise and high-throughput quantitative analysis of multiple targets.First,atomic force microscope(AFM)was used to verify the efficient synthesis of tFNA.The signal linear amplification capability of the reporter unit was verified by polyacrylamide gel electrophoresis(PAGE)and total internal reflection fluorescent microscope(TIRFM).The performance of the sensing interface substrates was compared,and the golden island chip with signal amplification was selected.The specificity of the detection system was verified by an interface specificity experiment.Five bladder cancer-related miRNAs were selected to construct standard curves for quantitative detection.Results·The efficient synthesis of tetrahedral monomer and dimer structures was verified by AFM.PAGE and TIRFM characterization verified the linear amplification of fluorescence signals from 1 to 6 valence fluorescence reporter units.In order to achieve further signal amplification,the plasma island chip and the traditional glass chip were compared.The results showed that the gold island chip exhibited a plasmonic effect,which significantly enhanced the near-infrared(NIR)fluorescence,with a signal amplification of up to 13.6 times compared to the glass chip.The specificity verification experiment showed that the signal-to-noise ratio of the system ranged from 7 to 10,demonstrating high specificity.Based on the high specificity of the system,along with the good interface regulation ability and linear amplification of the framework nucleic acid-based interface,dual-color parallel detection of the targets was finally realized.The working range was 100 fmol/L-10 nmol/L(R2≥0.991),and the detection limit was as low as 100 fmol/L.Conclusion·The establishment of this platform opens new avenues for highly sensitive quantitative analysis of biomarkers.Furthermore,the developed framework nucleic acid-based detection platform holds great potential for clinical diagnosis and prognosis of bladder cancer and other major diseases.Through early detection and precise subtype diagnosis,doctors can formulate more personalized treatment plans for patients,improving treatment efficacy and reducing unnecessary treatment plans and associated side effects.Therefore,this liquid biopsy technology not only provides new possibilities for early screening of bladder cancer but also serves as reference for research and clinical applications in other types of cancer.

Objective·To construct a framework nucleic acid-based linear amplification platform for the sensitive and quantitative detection of bladder cancer-related microRNAs(miRNAs),facilitating early screening and accurate diagnosis of bladder cancer.Methods·This study combined a plasma fluorescence-enhanced chip with high-performance tetrahedral framework nucleic acid(tFNA)probes,targeting miRNAs as biomarkers,to construct a framework nucleic acid-based linear signal amplification platform for precise and high-throughput quantitative analysis of multiple targets.First,atomic force microscope(AFM)was used to verify the efficient synthesis of tFNA.The signal linear amplification capability of the reporter unit was verified by polyacrylamide gel electrophoresis(PAGE)and total internal reflection fluorescent microscope(TIRFM).The performance of the sensing interface substrates was compared,and the golden island chip with signal amplification was selected.The specificity of the detection system was verified by an interface specificity experiment.Five bladder cancer-related miRNAs were selected to construct standard curves for quantitative detection.Results·The efficient synthesis of tetrahedral monomer and dimer structures was verified by AFM.PAGE and TIRFM characterization verified the linear amplification of fluorescence signals from 1 to 6 valence fluorescence reporter units.In order to achieve further signal amplification,the plasma island chip and the traditional glass chip were compared.The results showed that the gold island chip exhibited a plasmonic effect,which significantly enhanced the near-infrared(NIR)fluorescence,with a signal amplification of up to 13.6 times compared to the glass chip.The specificity verification experiment showed that the signal-to-noise ratio of the system ranged from 7 to 10,demonstrating high specificity.Based on the high specificity of the system,along with the good interface regulation ability and linear amplification of the framework nucleic acid-based interface,dual-color parallel detection of the targets was finally realized.The working range was 100 fmol/L-10 nmol/L(R2≥0.991),and the detection limit was as low as 100 fmol/L.Conclusion·The establishment of this platform opens new avenues for highly sensitive quantitative analysis of biomarkers.Furthermore,the developed framework nucleic acid-based detection platform holds great potential for clinical diagnosis and prognosis of bladder cancer and other major diseases.Through early detection and precise subtype diagnosis,doctors can formulate more personalized treatment plans for patients,improving treatment efficacy and reducing unnecessary treatment plans and associated side effects.Therefore,this liquid biopsy technology not only provides new possibilities for early screening of bladder cancer but also serves as reference for research and clinical applications in other types of cancer.

Objective To investigate the value of endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) in the diagnosis of sarcoidosis.Methods Twenty-two cases of clinically suspected sarcoidosis underwent bronchoscopy and ultrasound bronchoscopy examination,including endobronchial biopsy (EBB),transbronchial lung biopsy (TBLB),and EBUS-TBNA.The biopsy samples of EBB,TBLB,and EBUS-TBNA were paraffin-embedded for hematoxylin eosin (HE) staining and acid-fast staining,respectively.Differences in the diagnosis of sarcoidosis were compared among EBB,TBLB,and EBUS-TBNA.The diagnostic performance of EBUS-TBNA was evaluated.Interventional pulmonology diagnostic strategy of sarcoidosis was analyzed.Results Among all the 22 patients with suspected sarcoidosis,20 cases were diagnosed as sarcoidosis,1 case was small cell lung cancer,and 1 case was lymphoma.The number of patients who were diagnosed by EBB,TBLB,and EBUS-TBNA was 6 cases,9 cases,and 16 cases,respectively;and their diagnostic yield was 30.0％,45.0％,and 80.0％,respectively.The diagnostic yield of EBUS-TBNA was significantly better than the other two (P =0.005).Combined EBB and EBUS-TBNA,the diagnostic yield was 85.0％.Combined TBLB and EBUS-TBNA,the diagnostic yield was 90.0％.Combined those three,the diagnostic yield was 95.0％.EBUS-TBNA diagnostic yield was affected by the location and size of lymph nodes.The diagnostic yield of subcarinal lymph nodes and paratracheal lymph nodes by EBUS-TBNA was significantly better than that of Hilar lymph nodes (x2 =4.29,P ＜0.05),EBUS-TBNA showed better diagnostic yield for the lymph nodes whose diameter was greater than 2cm (x2 =4.067,P ＜ 0.05).EBUS-TBNA had fewer complications.Most patients only had a little bleeding in puncture site.Conclusions EBUS-TBNA contributed to diagnose sarcoidosis,while TBLB and EBB had a complementary value in the diagnosis of sarcoidosis by EBUS-TBNA.

Objective To establish a simultaneous method for determination of 5 constituents from Panax quinquefolium stem leaf 20(S)-protopanaxdiol saponins,Ginsenoside-Rb1,Rc,Rb2,Rb3 and Rd by RP-HPLC.MethodsPanax quinquefolium stem leaf 20(S)-protopanaxdiol saponins was provided by the Ji'an Yisheng Pharmaceutical Stock Co.Ltd..The Agilent ODS column(5 ?m,4.6 mm?250 mm)was used,and eluted with acetonitrile-0.2% phosphoric acid water in gradient mode.The UV detection wavelength was 203 nm,and the flow rate was 1.0 ml/min,with the temperature of column at 40 ℃.The accuracy,stability and repeability of this method was evaluated.The yield rate was measured.ResultsFive compounds were baseline separated,with good linearity.A standard curve was plotted.Our method was proved to have sound accuracy,stability and repeability.The yield rate was 98.39%,97.36%,98.02%,97.70% and 97.01% respectively for Ginsenoside-Rb1,Rc,Rb2,Rb3 and Rd.ConclusionOur method of determination is rapid and accurate,and can be used for controlling the quality of Panax quinquefolium 20(S)-protopanaxdiol saponins.