Background and purpose:The plasma used for routine coagulation test(CCT)can only reflect a single component at a certain coagulation time point/segment,while thromboelastography(TEG)can depict the overall dynamic process curve of coagulation and fibrinolysis,which can more independently and completely reflect the true state of the blood and can serve as a supplement to coagulation function testing.This study aimed to evaluate the application value of combined coagulation function indexes in monitoring the hypercoagulable state of patients with colorectal cancer after chemotherapy,and to explore the risk factors of thrombosis in patients with colorectal cancer after chemotherapy,so as to provide reference for clinical monitoring of hypercoagulable state.Methods:A total of 160 patients with colorectal cancer from Fudan University Shanghai Cancer Center from June 2021 to June 2023 were selected as the experimental group,and 80 healthy subjects were selected as the control group.Then the experimental group was divided into a group without thrombosis(82 cases)and a group with thrombosis(78 cases)according to whether they had thrombosis or not.The determinations of thromboelastography(TEG)[coagulation reaction time(R),coagulation formation time(K),blood clot formation rate(α-Angle),maximum amplitude(MA)and coagulation index(CI)],conventional coagulation tests(CCT)[activated partial thromboplastin time(APTT),prothrombin time(PT),thrombin time(TT),fibrinogen(Fib),D-dimer(DD),fibrinogen degradation products(FDP)]and platelet count(PLT)were studied among three groups.With or without thrombosis as the criterion of hypercoagulable state,statistically significant indicators were selected to be included in the binary logistic regression analysis,and the receiver operating characteristic(ROC)curve was drawn to analyze the diagnostic efficacy of single and combined detection of the coagulation function indicators for hypercoagulable state in patients with colorectal cancer after chemotherapy.Basic information,tumor stage and Autar score of deep vein thrombosis were collected in 160 patients with colorectal cancer.Logistic regression analysis was performed to explore the risk factors of thrombosis.This study was approved by the Ethics Committee of Fudan University Shanghai Cancer Center(number:050432-4-2108*).Results:Compared with the control group,the R,TT and PLT of the group with thrombosis were decreased(P＜0.05),while APTT,PT,DD and FDP were increased(P＜0.05).The differences in various indicators between the group with thrombosis and the control group were statistically significant(P＜0.05).Compared with the group without thrombosis,the K in the group with thrombosis decreased(P＜0.05),while Angle,MA,CI,FIB,DD and FDP all increased(P＜0.05).ROC curve analysis showed that in the assessment of hypercoagulable state in patients with colorectal cancer after chemotherapy,the area under curve(AUC)of TEG was 0.756,sensitivity was 67.5%,and specificity was 73.8%.The AUC of CCT was 0.691,sensitivity was 78.8%,and specificity was 56.2%.The combined detection AUC was 0.840,sensitivity was 80.0%,and specificity was 77.5%.In the analysis of risk factors,tumor stage,distant metastasis and Autar score were correlated with thrombus formation in patients with colorectal cancer after chemotherapy(P＜0.05),and the differences of the three risk factors in K,Angle,MA,CI,Fib,DD and FDP were statistically significant(P＜0.05).Conclusion:K,Angle,MA,CI,Fib,DD and FDP are the main indicators to reflect the hypercoagulable state,and the combined detection of TEG and CCT can better reflect the coagulation state of patients with colorectal cancer after chemotherapy.Tumor stage Ⅲ to Ⅳ,distant metastasis and high Autar score are risk factors for thrombosis.The incidence of thrombosis can be reduced by monitoring the relevant coagulation indicators in the high-risk population.

Objective:To investigate the association between tumor necrosis factor-β (TNF-β) gene polymorphisms and genetic predisposition to gastric cancer, and to analyze the relationship between specific genotype of TNF-β and serum levels of TNF-β.Methods:Using case control study, we selected 153 patients with gastric cancer in Fudan University Shanghai Cancer Center between September 2021 and December 2022 as the gastric cancer group, and 150 healthy individuals were chosen as the healthy control group. In the previous study, 30 peripheral blood DNA samples of gastric cancer patients and healthy controls respectively were amplified by conventional PCR, which were sequenced to identify the genotype frequencies of TNF-β polymorphic loci (rs1041981, rs2229092, rs2229094 and rs78613290); consequently, Allele-Specific Quantitative PCR was used to further detect and analyze the genotype and genotype frequencies of TNF-β polymorphic loci; serum TNF-β levels were measured by Enzyme-Linked Immunosorbent Assay (ELISA), and the relationship with specific genotypes of TNF-β was analyzed. Chi-square test and Fisher test were used to analyze the genotype distribution frequency of TNF-β polymorphic loci, and non-parametric statistics was used to analyze the differences in serum TNF-β expression levels.Results:The sequencing results showed that the genotype distribution of rs1041981 in gastric cancer group was CC 16.67% (5/30), CA 40.00% (12/30) and AA 43.33% (13/30). The genotype distribution in control group was CC 40.0% (12/30), CA 43.33% (13/30), AA 16.7% (5/30). The difference of genotype frequency between the two groups was statistically significant (χ 2=6.478, P=0.039). The genotypes of the polymorphic loci rs2229092 in both groups were AA, AG, and GG, with no statistically significant difference between the two groups (χ 2=1.888, P=0.612). The distribution frequencies of the genotypes of the polymorphic loci rs2229094 (TT and TC) and rs78613290 (GG and AG) showed no statistically significant differences between the two groups (both P>0.05). Further validation with an expanded clinical samples (153 cases in the gastric cancer group and 150 cases in the control group) found that the difference of rs1041981 genotype distribution between the gastric cancer group [CC 15.69%(24/153), CA 54.9%(84/153), AA 29.4%(45/153)] and the control group [CC 27.3%(41/150), CA 58.0%(87/150), AA 14.7%(22/150)] was significantly different (χ 2=12.366, P=0.002). Analysis of the influence of different allele frequencies on the risk of gastric cancer revealed that the odds ratio ( OR) of the A allele of rs1041981 for the risk of gastric cancer compared to the C allele was 1.701 (95% CI 1.235?2.355). Gene phenotype analysis combining the clinicopathological characteristics of gastric cancer patients found that the distribution frequency of the rs1041981 genotype was significantly different among groups of different genders, tumor invasion depths, and the lymph node metastasis, with statistically significant differences (All P>0.05). Additionally, gastric cancer patients with rs1041981 AA genotypes had higher serum TNF-β expression levels than those with CA and CC genotypes, (both P<0.05). Conclusions:The gene type frequency of the TNF-β gene polymorphic loci (rs1041981, C>A) exhibited significant differences between the gastric cancer group and the healthy control group. The presence of the A allele in rs1041981 site increased the susceptibility to gastric cancer, and patients with different gene types displayed vaning levels of serum TNF-β, among which AA genotype ranks the highest level.

Objective:To establish a risk assessment model for recurrence and metastasis in patients with advanced nasopharyngeal carcinoma.Methods:A survival follow-up study was conducted using a COX regression model to analyze 242 patients with advanced nasopharyngeal carcinoma who were treated for the first time in the Fudan University Shanghai Cancer Center from March 1, 2012 to August 31, 2020. The mean age was (48.33±11.13) years, with 178 males and 64 females. The mean survival was (3.39±1.42) years. According to the random number table method, the enrolled subjects were divided into two groups, including 192 cases in the modeling group and 50 cases in the validation group. Venous blood was collected from patients before treatment, after the first treatment and during the follow-up period after treatment. The blood cell classification and blood biochemical indicators were analyzed. T test and Chi-square test were used to analyze the difference in indicators in prognosis of patients with recurrence and metastasis as the outcome of the study. Multivariate COX regression analysis was used to screen out the independent prognostic factors affecting the recurrence and metastasis of nasopharyngeal carcinoma patients, and the Nomogram models of recurrence and metastasis risk of patients in 2 years, 4 years and 6 years were constructed. The model C-Index of the modeling group and the validation group were calculated to evaluate the performance of the predictive model.Results:White blood cells ( P=0.028), lymphocyte counts ( P<0.001), neutrophils ( P=0.001), platelets ( P=0.046), albumin ( P<0.001), neutrophil/lymphocyte ratio ( P<0.001), platelet/lymphocyte ratio ( P<0.001), lymphocyte/monocyte ratio ( P<0.001), systemic immune inflammatory response index ( P<0.001), systemic inflammatory response index ( P<0.001), and prognostic nutritional index ( P=0.004) had statistically significant differences in the efficacy monitoring of patients; through multivariate COX regression analysis, it was found that the platelet/lymphocyte ratio ( HR 2.537, 95% CI 1.439-4.473) and the prognostic nutritional index ( HR 0.462, 95% CI 0.236-0.903) are important factors to predict the risk of recurrence and metastasis of patients. Combining the above indicators, the Nomogram risk assessment model was established. The C index of the modeling group was 0.698, and the C index of the validation group was 0.739. The calibration curves of the two groups showed good consistency. Conclusion:The Nomogram evaluation model can accurately predict the risk of recurrence and metastasis in patients with nasopharyngeal carcinoma, and provide a theoretical basis for evaluating the prognosis of clinical treatment.

Liquid biopsy can non-invasively reveal the status of tumors and its prognosis in vivo, which is hotspots and difficulties of research in current era. Among them, an emerging marker called circulating tumor RNA (ctRNA) reflects the genetic information of tumor origin and provides a powerful basis for early diagnosis, targeted drug monitoring and prognosis prediction. At this stage, several ctRNA kits have been approved for clinical use. For example, microRNA (miRNA) can be used for aided diagnosis of liver cancer. And many transformation applications of promising ctRNA have been vigorously carried out. Despite the facts that there are still many clinical challenges of ctRNA detection technology to be solved effectively, ctRNA, as a new member of the liquid biopsy technology, has shown remarkable clinical value. Along with the mechanism becoming gradually clear, ctRNA will be a reliable diagnostic tool with the increasing clinical requirements for facilitating the tumor management.

Objective:To discuss the diagnostic value of calcitonin(CT), carcinoembryonic antigen (CEA), neuron-specific enolase (NSE), pro-gastrin releasing peptide (Pro-GRP) and chromogranin A (CgA) in the identification of medullary thyroid carcinoma (MTC).Methods:The CT levels in 105 cases of MTC, 50 cases of papillary thyroid carcinoma, 10 cases of thyroid follicular carcinoma, 5 cases of undifferentiated thyroid carcinoma, 50 cases of benign thyroid diseases, 30 cases of non-thyroid malignant tumors and 50 cases of healthy controls were measured from February 2017 to August 2019 at the Department of Clinical Laboratory, Cancer Hospital affliated to Fudan University. Additionally, 79 cases of MTC, 30 cases of non-MTC thyroid malignant tumors and 30 healthy controls were selected for the measurement of CEA, NSE, Pro-GRP and CgA levels. The receiver operating curve was utilized to clarify the area under the curve (AUC), sensitivity, and specificity of each indicator to distinguish between different groups.Results:The medians of CT concentrations in the group of MTC patients was 607.2 (152.5,2 777.5)pg/ml, which was statistically significantly higher than that of the subjects in the group of papillary thyroid carcinoma 1.48 (0.5,2.91)pg/ml, follicular thyroid carcinoma 1.90 (0.82,2.99)pg/ml, undifferentiated thyroid carcinoma 0.50 (0.50,4.93)pg/ml, benign thyroid disease 1.30 (0.50,2.79)pg/ml, non-thyroid malignancies 1.36 (0.50,2.89)pg/ml and healthy controls 2.05 (0.89,3.18)pg/ml. The sensitivity, specificity and AUC of CT to distinguish MTC vs. non-MTC patients was 96.2%, 99.3% and 0.99, respectively. The maximum diameter (>1 cm, P=0.001, OR=15.74) and number (>1, P=0.04, OR=3.4) of nodules were two independent risk factors for elevated CT. CEA (AUC=0.94), NSE (AUC=0.65), Pro-GRP (AUC=0.94) and CgA (AUC=0.83) could all distinguish MTC vs. non-MTC thyroid malignancies. The AUC, sensitivity and specificity by combining CT, CEA, NSE, Pro-GRP and CgA to differentiate MTC vs. non-MTC thyroid malignancies was 1, 100% and 100%, respectively. Conclusions:CT, CEA, NSE, Pro-GRP and CgA may be helpful for the auxiliary diagnosis of MTC. The combination of these indicators in the diagnosis of MTC has high sensitivity and specificity.

Objective:To discuss the diagnostic value of calcitonin(CT), carcinoembryonic antigen (CEA), neuron-specific enolase (NSE), pro-gastrin releasing peptide (Pro-GRP) and chromogranin A (CgA) in the identification of medullary thyroid carcinoma (MTC).Methods:The CT levels in 105 cases of MTC, 50 cases of papillary thyroid carcinoma, 10 cases of thyroid follicular carcinoma, 5 cases of undifferentiated thyroid carcinoma, 50 cases of benign thyroid diseases, 30 cases of non-thyroid malignant tumors and 50 cases of healthy controls were measured from February 2017 to August 2019 at the Department of Clinical Laboratory, Cancer Hospital affliated to Fudan University. Additionally, 79 cases of MTC, 30 cases of non-MTC thyroid malignant tumors and 30 healthy controls were selected for the measurement of CEA, NSE, Pro-GRP and CgA levels. The receiver operating curve was utilized to clarify the area under the curve (AUC), sensitivity, and specificity of each indicator to distinguish between different groups.Results:The medians of CT concentrations in the group of MTC patients was 607.2 (152.5,2 777.5)pg/ml, which was statistically significantly higher than that of the subjects in the group of papillary thyroid carcinoma 1.48 (0.5,2.91)pg/ml, follicular thyroid carcinoma 1.90 (0.82,2.99)pg/ml, undifferentiated thyroid carcinoma 0.50 (0.50,4.93)pg/ml, benign thyroid disease 1.30 (0.50,2.79)pg/ml, non-thyroid malignancies 1.36 (0.50,2.89)pg/ml and healthy controls 2.05 (0.89,3.18)pg/ml. The sensitivity, specificity and AUC of CT to distinguish MTC vs. non-MTC patients was 96.2%, 99.3% and 0.99, respectively. The maximum diameter (>1 cm, P=0.001, OR=15.74) and number (>1, P=0.04, OR=3.4) of nodules were two independent risk factors for elevated CT. CEA (AUC=0.94), NSE (AUC=0.65), Pro-GRP (AUC=0.94) and CgA (AUC=0.83) could all distinguish MTC vs. non-MTC thyroid malignancies. The AUC, sensitivity and specificity by combining CT, CEA, NSE, Pro-GRP and CgA to differentiate MTC vs. non-MTC thyroid malignancies was 1, 100% and 100%, respectively. Conclusions:CT, CEA, NSE, Pro-GRP and CgA may be helpful for the auxiliary diagnosis of MTC. The combination of these indicators in the diagnosis of MTC has high sensitivity and specificity.

Liquid biopsy can non-invasively reveal the status of tumors in vivo, and provides a powerful basis for early diagnosis, personalized treatment monitoring and prognosis prediction. According to the type of tumor-associated material, liquid biopsy covers circulating tumor cells (CTCs), circulating tumor DNA (ctDNA), extracellular vesicles (EVs) and circulating tumor RNA (ctRNA) etc. At present, several liquid biopsy products havebeen approved for clinical use, while many transformation studies have been vigorously carried out. However, there are still many clinical challenges to be solved effectively. Despite the standardization of detection and quality management system of liquid biopsy are lagged with the rapid development of technology, liquid biopsy, as a highly promising detection technology, will become a reliable diagnostic tool with the increasing clinical requirements for facilitating the tumor management.

Objective:To investigate the clinical value of glypican-3 (GPC3) detection in the diagnosis and therapy-monitoring of primary hepatocellular carcinoma (HCC).Methods:From March 2018 to May 2019, the patients with HCC were enrolled as the experimental group( n=166)from Fudan University Shanghai Cancer Centre, while the specimens from health control group( n=94) and benign control group ( n=50) were analyzed. The serum of GPC3 and alpha fetoprotein (AFP)levels were respectively detected by ELISA and chemiluminescence. GPC3 detections combined with AFP etc. in accuracy of HCC diagnosis were explored by using Logistic regression analysis. Results:The serum GPC3 level in patients with HCC ［0.210 (0.048, 0.801)mg/L］ ［Median (quartile Q1, Q3)］ was significantly higher than those in healthy controls ［0.029(0.019, 0.052)mg/L］ and benign controls ［0.033(0.021, 0.043) mg/L］ ( Z=-7.69, P<0.001).The serum levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT) and AFP were significantly different among the three groups ( Z=-7.02, -6.85, -8.36 respectively, P<0.001). Among the serological indicators, it was related to ALT and AST ( Z=-3.77, -4.09 respectively, P<0.001).The Cut-off level of GPC3 was determined as 0.077 mg/L by ROC curve. The sensitivity of the combined detection of serum GPC3 with AFP for HCC was up to 87.82%, the specificity was 77.86%, the negative predictive value was 84.29%, and the positive predictive value was 82.53%.The HCC-GPC3 model was constructed by using Logistic regression analysis. The area under the ROC curve was 0.882, the total sensitivity was 91.10%, and the total specificity was 72.73%. Further analysis showed that the serum GPC3 of patients with HCC was significantly lower ［0.454(0.019, 0.286) mg/L］ than that before surgery［0.608(0.039, 0.554）mg/L］( Z=-7.32, P<0.001). Conclusion:The detection of serum GPC3 can be applied to aid diagnosis and therapy-monitoring of HCC.The combination of GPC3 and AFP can improve the diagnostic efficiency of HCC.

Objective:To investigate the clinical value of glypican-3 (GPC3) detection in the diagnosis and therapy-monitoring of primary hepatocellular carcinoma (HCC).Methods:From March 2018 to May 2019, the patients with HCC were enrolled as the experimental group( n=166)from Fudan University Shanghai Cancer Centre, while the specimens from health control group( n=94) and benign control group ( n=50) were analyzed. The serum of GPC3 and alpha fetoprotein (AFP)levels were respectively detected by ELISA and chemiluminescence. GPC3 detections combined with AFP etc. in accuracy of HCC diagnosis were explored by using Logistic regression analysis. Results:The serum GPC3 level in patients with HCC ［0.210 (0.048, 0.801)mg/L］ ［Median (quartile Q1, Q3)］ was significantly higher than those in healthy controls ［0.029(0.019, 0.052)mg/L］ and benign controls ［0.033(0.021, 0.043) mg/L］ ( Z=-7.69, P<0.001).The serum levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT) and AFP were significantly different among the three groups ( Z=-7.02, -6.85, -8.36 respectively, P<0.001). Among the serological indicators, it was related to ALT and AST ( Z=-3.77, -4.09 respectively, P<0.001).The Cut-off level of GPC3 was determined as 0.077 mg/L by ROC curve. The sensitivity of the combined detection of serum GPC3 with AFP for HCC was up to 87.82%, the specificity was 77.86%, the negative predictive value was 84.29%, and the positive predictive value was 82.53%.The HCC-GPC3 model was constructed by using Logistic regression analysis. The area under the ROC curve was 0.882, the total sensitivity was 91.10%, and the total specificity was 72.73%. Further analysis showed that the serum GPC3 of patients with HCC was significantly lower ［0.454(0.019, 0.286) mg/L］ than that before surgery［0.608(0.039, 0.554）mg/L］( Z=-7.32, P<0.001). Conclusion:The detection of serum GPC3 can be applied to aid diagnosis and therapy-monitoring of HCC.The combination of GPC3 and AFP can improve the diagnostic efficiency of HCC.

Single tumor marker(TM)for cancer diagnosis with both satisfactory sensitivity and specificity has not been found.Therefore, it is necessary to identify the biomarkers to make the TM panel and establish the corresponding multiparameter predictive model through fuzzy mathematics methods.Recent studies using the big data technology showed that the integration of the biomarkers panel and prediction score model could provide the effective and accurate diagnosis for the cancer patients and be in favor of making the scheme for personalized therapy,treatment-monitoring and prognosis prediction.A reliable and efficient TM score prediction model will provide the novel perspectives and procedures for the diagnosis and treatment of cancer patients.