| Background:At present,the incidence rate of thyroid cancer is increasing year by year,and it has become the most common endocrine system malignant tumor in the world.The clinical guidelines for the diagnosis and treatment of thyroid cancer at home and abroad have become increasingly mature,especially for differentiated thyroid cancer,which has a standardized clinical diagnosis and treatment pathway.At present,the diagnosis of thyroid cancer mainly relies on ultrasound imaging and fine needle aspiration biopsy(FNAB)guided by ultrasound.However,this invasive examination to some extent reduces patient compliance and has limitations in the inability to puncture due to the special location of the nodule.At the same time,although the highest incidence rate of thyroid papillary carcinoma(PTC)has a mature individual refinement program of surgical treatment,endocrine suppression treatment and radiotherapy,its characteristics of lymph node metastasis still make PTC metastasis and recurrence risk higher.In recent years,some scholars have proposed using preoperative serum thyroglobulin(Tg)levels to evaluate the properties of thyroid nodules.However,there is still controversy over whether preoperative serum Tg levels are suitable for the evaluation of newly diagnosed differentiated thyroid cancer patients.Therefore,serum Tg levels are currently only used as tumor markers for evaluating the recurrence of differentiated thyroid cancer in clinical practice.In addition to the diagnosis of thyroid nodules,accurate and effective prognostic assessment is also important for thyroid cancer.At present,relevant clinical examination items have been routinely used for follow-up and treatment planning after thyroidectomy,including ultrasound examination,serum thyroid function,and Tg.In addition,recent studies have shown that inflammatory markers in peripheral blood,such as C-reactive protein(CRP),can also be used for postoperative evaluation of thyroid cancer.When thyroid autoantibodies(Tg Ab)are present,the serum Tg level will be reduced,affecting the accuracy of monitoring the condition through Tg detection.At this time,serum CRP levels can be simultaneously detected to predict the postoperative disease status of patients.However,there is still some controversy surrounding these prognostic assessment methods,and more research is needed to further validate their role in practical clinical applications.In summary,this study proposes to apply real-time monitoring of serum CRP and serum Tg levels to the comprehensive diagnosis and treatment system of differentiated thyroid cancer.1)Statistical analysis was conducted on the serum CRP,Tg,and Tg Ab levels of patients with PTC in different groups before and after total thyroidectomy,to evaluate the significance of these serological indicators for the prognosis of PTC.2)Introduce the concept of Point of Care Testing(POCT)into the diagnosis and treatment of thyroid cancer.By utilizing the advantages of simple operation,low cost,and rapid detection of lateral flow immunoassay(LFIA)test strips,a gold nanorod(GNR)modified LFIA test strip was established to achieve real-time and rapid bedside detection of Tg,providing reference for timely detection of metastatic lesions before surgery and guiding surgical plan selection during surgery.3)Establishing a bedside LFIA detection method for whole blood CRP and Tg,enabling patients to perform CPR and Tg levels at home,enabling patients with inconvenient medical treatment to promptly detect inflammatory diseases or recurrence and metastasis,reducing the economic burden and psychological pressure of patients visiting hospitals for follow-up examinations,and providing new methods and ideas for the early diagnosis,prognosis evaluation,and disease management of differentiated thyroid cancer.Objective:1.Through retrospective analysis of clinical data of PTC patients,explore the application value of CRP and Tg levels in the diagnosis and treatment of PTC.2.Synthesize GNR materials with both colorimetric and photothermal effects.3.Prepare GNR modified with anti CRP antibodies as nanoprobes,and construct GNR labeled LFIA test strips using a dual antibody sandwich method to establish a portable detection method with efficient and stable CRP rapid on-site detection capability.4.Prepare GNR modified with anti Tg antibodies as nanoprobes,and construct GNR labeled LFIA test strips using dual antibody sandwich method to establish a portable detection method with efficient and stable Tg rapid on-site detection capability.Methods:1.Firstly,a retrospective analysis was conducted on the clinical data of 180 PTC patients admitted to the First Hospital of Jilin University from December 2017 to December 2023.Compare the relationship between preoperative serum CRP,Tg,Tg Ab levels and clinical pathological characteristics of PTC patients through statistical analysis.Secondly,a retrospective analysis was conducted on the clinical data of 230 patients with thyroid diseases admitted to the First Hospital of Jilin University from December 2013 to December 2023.Among them,the experimental group consisted of 200 PTC patients awaiting treatment with radioactive iodine 131,while the control group consisted of 30 patients with Hashimoto’s thyroiditis(HT).Among these 200 postoperative PTC patients,there are 150 pure PTC patients and 50 PTC combined with HT patients.Organize the clinical and pathological data of these230 patients,and conduct statistical analysis using SPSS 27.0 software to compare the relationship between postoperative serum CRP and Tg levels and the prognosis of PTC patients,in order to explore the application value of CRP and Tg in the diagnosis and treatment of PTC.2.Synthesize GNR using seed growth method and characterize it.Use 650 nm laser irradiation to evaluate the photothermal conversion capability of GNR and calculate the photothermal conversion efficiency of the synthesized GNR.3.Prepare dual-mode LFIA test strips with both colorimetric and photothermal effects using synthesized GNR.Using GNR modified with anti CRP antibodies as colorimetric and photothermal conversion probes.Mouse anti CRP monoclonal antibody and goat anti mouse Ig G were used as detection and quality control lines,respectively.Using the classic dual antibody immune sandwich method,antibody modified GNR was introduced as a recognition probe to construct a dual-mode side flow immunochromatographic strip capable of rapidly detecting CRP.By directly comparing colors and using a handheld thermal imager to read signals,the detection results can be directly observed by the naked eye.Image J and thermal imagers can also be used to collect chromaticity and temperature signals for further quantitative results.Applying this method to analyze serum spiked samples,the same sample is tested three times to calculate its mean and standard deviation,and its sensitivity,specificity,anti-interference ability,and storage stability are examined.Further testing of actual clinical samples is conducted to evaluate the applicability of the prepared FLIA test strip for CRP detection in clinical practice,and its accuracy is verified by comparing it with diagnostic kits used in clinical practice.4.Referring to the above CRP detection test strip establishment process,prepare a dual-mode LFIA test strip with GNR labeling for rapid detection of Tg.Signal reading through direct colorimetry and handheld thermal imager: The detection results can be directly observed by the naked eye,or quantitative detection can be achieved by monitoring colorimetry and temperature signals using Image J and thermal imager.Apply the LFIA test strip to analyze serum spiked samples and examine its sensitivity,specificity,anti-interference ability,and storage stability.At the same time,further testing will be conducted on actual clinical samples(including serum samples,whole blood samples,and lymph node puncture eluent samples)to evaluate their applicability for Tg detection in actual clinical practice.Using Roche chemiluminescence as a control and pathological examination as the gold standard to verify its accuracy.Results:1.There was no difference in preoperative CRP,Tg,and Tg Ab levels among PTC patients of different ages and genders(P>0.05).The preoperative serum CRP and Tg Ab levels in patients with multiple lesions were significantly higher than those in the single lesion group,while Tg levels were lower(P<0.05).The preoperative serum CRP and Tg levels in PTC patients with extraglandular invasion,lymph node metastasis,or larger tumors(diameter>2 cm)were higher than those without extraglandular invasion,lymph node metastasis,or smaller tumors(diameter ≤ 2 cm)(P<0.05).Through ROC curve analysis,it was found that preoperative serum CRP and Tg levels have high diagnostic value for extraglandular invasion and lymph node metastasis of PTC.2.The comparison between the HT group,HT+PTC group,and PTC group concluded that there was no statistically significant difference in gender among the groups(P>0.05),but there was a statistically significant difference in age(P>0.05).There was no statistically significant difference(P>0.05)in the prognosis(recurrence/disease persistence)between the HT+PTC group and the PTC group.The serum CRP and Tg levels of recurrent/persistent PTC patients were significantly higher than those of non recurrent patients(P<0.05).The serum CRP levels in patients with persistent/recurrent PTC in the HT+PTC group were significantly higher than those in patients with HT or PTC(with or without recurrence)(P<0.05).There is a positive correlation between serum CRP levels and Tg Ab levels in patients with recurrent/persistent PTC.Through ROC curve analysis,it was found that elevated serum CRP and Tg levels have high diagnostic value for recurrent/persistent PTC.The disease-free survival of PTC patients with Tg ≥ 39.21 ng/m L and CRP ≥ 6.33mg/L is significantly shorter than that of patients with Tg<39.21 ng/m L and CRP<6.33 mg/L.3.The synthesized GNR has a high photothermal conversion efficiency(η=39%)and strong local surface plasmon resonance(LSPR)absorption effect.4.The LFIA test strip we prepared for CRP detection has high specificity and sensitivity.In the range of 50 ng/m L to 10000 ng/m L,the concentration of CRP shows a good linear relationship with the T/C values of colorimetric and temperature signals,and exhibits a lower detection limit(LOD,1.3 ng/m L).5.This detection method can be successfully applied to the detection of CRP in clinical serum samples.Using the immunoturbidimetric diagnostic kit as a reference method,it was found that the T/C values of colorimetric and temperature signals showed a good linear relationship with the logarithm of CRP concentration.Comparing the preoperative serum CRP levels of patients with nodular goiter and PTC using two detection methods,the results showed that there was no significant difference in CRP levels between the two groups of patients,indicating good consistency in the detection results of the two methods.In the detection of whole blood samples from 200 PTC patients,it was found that the AUC value of the test strip for CRP detection was 0.981,with a sensitivity of 98.6% and a specificity of97.7%;The diagnostic accuracy of the test strip is 98.00%,with a true positive rate of98.61%,a true negative rate of 97.66%,a false positive rate of 2.34%,and a false negative rate of 1.39%.6.The LFIA test strip based on GNR for Tg detection has high specificity and sensitivity.In the range of 3.5 ng/m L to 88 ng/m L,the T/C values of colorimetric and temperature signals show a good linear relationship with the logarithm of Tg concentration,with a detection limit of 1.4 ng/m L.7.In the detection of serum samples from 40 patients with thyroid diseases,the Tg detection strip method established in this study showed good consistency with the chemiluminescence method of commercial reagent kits,with an accuracy of 95%.In the detection of whole blood samples from 200 PTC patients,it was found that the AUC value of the test strip for Tg detection was 0.946,with a sensitivity of 96.9%and a specificity of 92.3%;The diagnostic accuracy of this test strip is 96.00%,with a true positive rate of 96.89%,a true negative rate of 96.31%,a false positive rate of7.69%,and a false negative rate of 3.11%.In the Tg detection of lymph node puncture eluate,the sensitivity of this test strip is 91.6%,the specificity is 94.7%,and the corresponding AUC value is 0.932(95% CI: 88.81%~97.49%),indicating that this test strip has high diagnostic value for PTC lymph node metastasis.8.There was no statistically significant difference in preoperative Tg levels between patients with nodular goiter and PTC(P>0.05);The higher the preoperative Tg level,the more prone PTC is to extraglandular invasion or lymph node metastasis(P<0.05);The occurrence of lymph node metastasis in PTC is not related to gender or age(P>0.05).Conclusion:1.Preoperative detection of serum CRP and Tg levels has certain significance in predicting extraglandular invasion and lymph node metastasis of PTC.Serum CRP and Tg levels can serve as important indicators for evaluating the recurrence or persistence of PTC.When Tg Ab is positive,detecting CRP can better predict the prognosis of the disease.CRP and Tg have certain evaluation value for the disease-free survival of PTC patients.2.GNR with unique colorimetric and photothermal properties was synthesized using seed growth method.3.A dual-mode LFIA test strip for rapid CRP detection has been successfully developed based on GNR.This LFIA test strip has high specificity and sensitivity for CPR detection,with a wide detection range and low detection limit.It has good applicability in CRP detection of clinical patient serum and whole blood samples.4.A dual-mode LFIA test strip for rapid detection of Tg has been successfully developed based on GNR.This LFIA test strip has high specificity and sensitivity for Tg detection,with a wide detection range and low detection limit.It has good applicability in the detection of Tg in clinical patient serum,whole blood samples,and lymph node puncture eluate. |
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