Calcitonin participates in physiological regulation of calcium metabolism, but it might not be the key factor.Multiple immunoassay methods have been developed for serum calcitonin detection.However, significant differences exist among the methods, and the results of immunoassay are affected by many factors.This paper mainly discusses some major advances in experimental researches of calcitonin and the influence factors of immunological measurement.

Objective To evaluate the differences of serum TSH of suspicious subclinical hypothyroidism determined by four automatic biochemical analyzers and the impact on clinical diagnosis and treatment.Methods Taking results of Roche Cobas e601 laboratory test as a reference, 103 serum samples with TSH 2.50-10.00 mU/L(90 with TSH≥4.27 mU/L) and normal FT3, FT4 were selected.Four different automatic biochemical analyzers (Cobas e601, Immulite2000, Centaur XP, I2000) were used to measure TSH of the serum samples at the same time.Wilcoxon signed rank test, Spearman correlation analysis were used for data analysis.Results TSH (M(P25, P75)) measured by 4 methods were 5.20(4.73, 6.40), 2.95(2.59, 3.48), 3.30(2.94, 4.15) and 4.10(3.43, 4.75) mU/L, which varied significantly from one assay to another (z values:-8.78,-8.41,-7.64,-8.09,-8.50, all P<0.05).The correlations between methods were of great differences (rs ranged from 0.45 to 0.92).Significant differences existed in each other for subclinical hypothyroidism diagnosis based on TSH cutoff respectively.Conclusion Results from different automatic immunoassay analyzers in patients with TSH of 2.50-10.00 mU/L varied widely, hence, it is indeterminate to diagnose subclinical hypothyroidism only relies on a single serum TSH test.

Objective To compare the consistence and difference between the assay results of the second generation of Tg (Tg Ⅱ) and the first generation of Tg (Tg Ⅰ) immunoassay,as well as to evaluate the impact of Tg Ⅱ on the clinical management of thyroid diseases.Methods Serum samples of 249 patients (30 with benign thyroid disorders and 219 with DTC;64 males and 185 females,average age 43.0 years)were collected and assayed by Tg Ⅱ and TgⅠ kits simultaneously.The measuring ranges of TgⅠ and TgⅡ were 0.10-1 000.00 μg/L and 0.04-500.00 μg/L,respectively.Data were analyzed by the Wilcoxon rank sum test and Spearman correlation analysis using IBM SPSS 19.0.Results The assay results of TgⅡ and TgⅠ strongly correlated (rs =0.979,P＜0.05).However,the median value of TgⅡ (2.31 (0.06-13.17) μg/L) was lower than that of TgⅠ(3.63(0.41-16.84) μg/L)(z=-13.25,P＜0.001).The difference between Tg Ⅱ and Tg Ⅰ got bigger when TgⅠ value decreased more.TgⅡ values were 11.09％ lower than TgⅠ (5.61(1.07-26.39) μg/L) vs 6.31(2.07-33.93) μg/L;z=-4.78,P＜0.05) in 30 patients with benign thyroid disorders and 37.71％ lower (2.18(0.07-7.47) μ.g/L) vs 3.50(0.39-10.18) μg/L;z=-9.02,P＜0.001) in 108 DTC patients without 131 Ⅰ treatment.But the above changes had no influence on clinical diagnosis and treatment.In the 71 DTC patients post 131Ⅰ treatment with low TSH and normal TgAb,there were 3 cases with TgⅠ＞1.0 μg/L but TgⅡ＜1.0μg/L,and 12 cases with TgⅠ＞0.1 μg/L but TgⅡ＜0.04 μg/L.Conclusions Serum TgⅡand Tg Ⅰ assay results are strongly correlated,though Tg Ⅱ value is slightly lower than Tg Ⅰ value.This difference may have no significant influence on the clinical diagnosis of thyroid diseases.However,TgⅡ may be better to evaluate the curative effect in some DTC patients post 131Ⅰ therapy.

Objective To interpret the major characteristics of literatures cited by 2015 ATA management guidelines for adult patients with thyroid nodules and DTC (2015 version).Methods The titles,datelines of the references,the medical specialties and regional distribution of the journals,and the definition of the scientific evidence rating for relevant references were extracted and analyzed.The data were roughly compared with those of 2009 revised ATA management guidelines for patients with thyroid nodules and DTC (2009 version).Results A total of 1 078 literatures,from 172 journals and 8 books,were cited by 2015 version,with 63 years spacing from 1952 to 2015.Extensive medical specialties were involved.The journals were world-wide distributed but the regional bias was obvious.Compared to the 2009 version,2015version adopted more recent literatures,and used more evidence rating for the recommendations.However,references with high-quality evidence were both less than 50％ in the two versions.Conclusion Huge amount of references with multi-specialties have been cited in 2015 version,however the regional distribution bias is distinctive and references with high-quality evidence are still insufficient.

Objective To explore the feasibility of inhibitor of apoptosis protein 2-mucosa associated lymphoid tissue lymphoma translocation gene 1 (API2-MALT1) fusion gene detection in endoscopic biopsy tissue of primary gastric lymphoma (PGL), and to study the expression of this fusion gene in PGL and its clinical values in diagnosis and treatment of PGL.MethodsA total of 32 suspicious PGL patients underwent endoscopic ultrasonography (EUS) examination and mucosal biopsy.The biopsy specimens were conducted histopathology examination, immunohistochemistry detection and real time RT-PCR for API2-MALT1 fusion gene expression.The expression of API2MALT1 fusion gene in clearly diagnosed PGL and its relation with PGL diagnosis, classification and treatment were analyzed and summarized.ResultsA total of 14 cases of 32 suspicious PGL patients were diagnosed as PGL by histopathology and immunohistochemistry examination, which including 11 cases of gastric mucosa-associated lymphoid tissue (MALT) lymphoma and 3 cases of gastric diffuse large B-cell lymphoma (DLBCL).There were 5 API2-MALT1 fusion gene positive cases, which were all MALT lymphoma, about 5 out of total 11 MALT lymphoma patients.API2-MALT1 fusion gene expression was negative in all 3 gastric DLBCL cases.The depth of lesion invasion and lymph nodes metastasis were more severe in API2-MALT1 fusion gene negative group than those of positive group.There were 2 of 5 API2-MALT1 fusion gene positive cases were Hp positive, and 5 of 9 negative cases were Hp positive.The anti-Hp therapy in 5 API2-MALT1 fusion gene positive cases was ineffective,however, chemotherapy was effective.In negative group, 2 of 5 Hp positive cases was complete remission and 4 Hp negative cases were ineffective with anti-Hp therapy.ConcltsionsAPI2-MALT1 fusion gene is common genetic abnormality in gastric MALT lymphoma.The detection of this fusion gene expression in endoscopic biopsy specimen by real time RT-PCR is clinically practical, and the detection of this fusion gene is valuable in the assessment of PGL diagnosis, treatment and prognosis.

Objective To explore the diagnostic model and clinical application value of serum proteomic fingerprint in inflammatory bowel disease (IBD) .Methods Serum proteome profiles of 72 IBD patients (54 Crohn′s disease (CD) and 18 ulcerative colitis (UC) and 44 healthy controls were analyzed by the weak cation exchange (WCX) beads combined matrix‐assisted laser desorption/ionization time of flight mass spectrometry (MALDI‐TOF‐MS ) technique . Among three groups , every two groups were compared .Wilcoxon rank sum test was used to screen out the peaks of difference expressed protein (P<0 .05) .Genetic algorithm combining with support vector machine (SVM ) was utilized to select the best diagnostic model .The predictive effects of this model was evaluated by leave one out method (LOO ) . Results The 10 most discriminating protein peaks were screened out between CD group and healthy control group , between UC group and healthy control group , between CD group and UC group . A diagnostic model established with four protein peaks ,the mass‐to‐charge ratio (M /Z ) of them was 3 275 .29 ,4 963 .91 ,4 980 .53 and 5 336 .90 ,could better distinguish CD and healthy controls .The specificity was 97 .7% ,and the sensitivity was 92 .6% in CD diagnosis .A diagnostic model established with four protein peaks ,the M /Z of them was 2 272 .41 ,2 660 .42 ,3 029 .77 and 5 002 .78 ,could better distinguish UC and healthy controls .The specificity was 100 .0% ,and the sensitivity was 94 .4% .A specificity was 50 .0% and sensitivity was 88 .9% in CD diagnosis with the diagnostic model of six protein peaks and the M /Z of them was 2 082 .63 ,2 210 .64 ,4 039 .02 ,4 298 .30 ,4 978 .03 ,5 002 .22 .Conclusion The diagnostic model of serum difference expressed protein in CD and UC is established by MALDI‐TOF‐MS technique and genetic algorithm combining with SVM ,which has high diagnostic value in IBD .

Objective To investigate the clinical significance of serum anti-Saccharomyces cerevisias antibody (ASCA),anti-outer membrane porin C (anti-OmpC),antibody to Pseudomonas fluorescens-associated sequence I2 (anti-I2 )and antibody to bacterial flagellin (anti-CBirl )in the diagnosis and treatment of inflammatory bowel disease (IBD).Methods From 2011 to 2013,87 patients with IBD were enrolled and divided into Crohn′s disease (CD)group (66 cases)and ulcerative colitis (UC)group (21 cases).A total of 62 age and gender matched healthy individuals were enrolled as the control group. Fasting blood samples (2 mL)of the subjects were collected.The expression of ASCA,anti-OmpC,anti-I2 and anti-Cbirl antibodies was detected with enzyme-linked immunosorbent assay (ELISA)kits.The diagnosis value of each antibody in IBD and the differential diagnostic value of in UC and CD were compared by receiver operating characteristic (ROC)curve.Results The area under the curve (AUC)of ASCA between IBD and the healthy control group,between CD group and UC group was 0.580 and 0.512, respectively;the accuracy in diagnosis was low.The AUC of anti-CBirl between IBD and the healthy control group was 0.617.There was no differential diagnosis significance of the other antibodies.The positive rate of ASCA in IBD group was 62.1 % (54/87),which was significantly higher than that in the control group (38.7%,24/62).The positive rates of anti-OmpC and anti-I2 in IBD group was significantly lower than those in the control group and the differences were statistically significant (both P <0.05).No difference was observed in positive rates of serum antibodies among the others groups (all P >0.05).The specificity,sensitivity,positive predictive value (PPV)and negative predictive value (NPV)of ASCA in differential diagnosis of CD and UC was 52.4%,66.7%,81 .48% and 33.33%,respectively.The specificity and sensitivity of anti-OmpC,anti-I2 and anti-CBirl in differential diagnosis of CD and UC was 81 .0% to 100.0% and 9.1 % to 37.9%,respectively.The specificity,sensitivity,PPV and NPV of double-positive ASCA and anti-I2 in the diagnosis of CD was 57.1 %,86.4%,82.6% and 50.0%, respectively.The positive rate of ASCA and anti-I2 in CD group was significantly higher than that in UC group (84.8%(56/66)vs 57.1 % (12/21 );χ2 =5 .633,P =0.018 ).Conclusions Positive ASCA has some significance in the diagnosis of patients with IBD in our country.The detection of anti-I2 can help to diagnose ASCA negative CD.Because of low sensitivity and positive rate,anti-OmpC and anti-CBirl have limited value in the diagnosis of IBD and the differential diagnosis of UC and CD in our country.

Objective To investigate the clinical and imaging characteristics of phosphaturic mesenchymal tumor and improve the clinical diagnosis. Methods From November 2014 to September 2017, 22 patients with pathologically confirmed diagnosis as phosphaturic mesenchymal tumor (PMT) were retrospectively analyzed, including 12 males and 10 females, age ranged from 30-72 years, mean (47 ± 11) years old. The clinical data, laboratory tests [serum calcium, phosphorus, alkaline phosphatase, parathyroid hormone and 1, 25- (OH) 2 D] and imaging examinations (X-ray, CT, MRI, nuclide) were collected and explored. Sixteen patients underwent SPECT scan and seven underwent PET/CT scan. Twenty patients had X-ray, eighteen patients had CT and 12 patients had MRI with enhancement. Results All patients suffered from diffuse pain for one to fifteen years, especially in lower back and lower extremities. All patients were found with low serum phosphorus, normal serum calcium. Twenty-one patients were found with elevated alkaline phosphatase, 16 with increased parathyroid hormone and 15 with decreased 1, 25 - (OH) 2 D. Thirteen lesions were located in the medullary cavity, seven in the soft tissue and two in the sinuses. Nineteen cases showed varying degrees of trabecular bone sparse, osteoporosis and osteomalacia on X-ray;There were 15 cases of multiple pseudo-fractures, including four cases of pelvic fracture complicated with femoral fracture, six cases of single fracture of pelvis, four cases of femur and one case of fibula. And seven cases showed multiple vertebral compression fractures. Thirteen lesions showed soft-tissue density and four in the medullary cavity showed high density on CT scan. The lesions presented low signal intensity on T1WI,high or low signal intensity on T2WI FLAIR and obviously enhanced in 12 patients who underwent MRI enhancement. Conclusion For patients with decreased serum phosphorus, elevated alkaline phosphatase, bone softening and fracture, octreotide or other nuclides should be primary imaging modality for confirming the location of the lesion. CT and MRI can further evaluate the nature of the lesion and improve diagnostic accuracy.

Objective: To screen and identify serum protein biomarkers for the differential diagnosis between ischemic colitis (IC) and ulcerative colitis (UC) by tandem mass tag (TMT) combined with liquid chromatography/tandem mass spectrometry (LC-MS/MS). Methods: From January 2018 to January 2019, at the First Affiliated Hospital of School of Medicine of Zhejiang University, patients with UC or IC, and health controls, each 10 cases, were enrolled into UC group, IC group and normal control (NC) group, respectively. Fasting serum samples of all the subjects were collected. After removal of high-abundance protein, followed by proteolysis, peptide labeling and fractionating, the samples were then processed by mass spectrometry. The protein with TMT data of three groups was obtained and protein with TMT value 0 were removed. Heat map of protein was constructed. The differential protein was defined as the protein fold change over 1.5 or less than 0.67. The Reactome database was used to cluster the pathways of differential proteins among groups. Statistical methods included t test, hypergeometry test and corrected by BH multiple test. Results: A total of 357 serum proteins were identified by proteomic profiling. There were 27 differential proteins between the IC group and the NC group, including six up-regulated proteins and 21 down-regulated proteins. There were 228 differential proteins between the UC group and the NC group, including 75 up-regulated proteins and 153 down-regulated proteins. There were 49 differential proteins between UC group and IC group, including 22 up-regulated proteins and 27 down-regulated proteins. In the comparison of differential proteins between the NC group, IC group and UC group, only the expression of fibrin 3 was statistically significant (the fold change between UC and NC, between UC and IC, between IC and NC were 0.24, 0.46 and 0.53, respectively; t=-5.089, -7.298 and -3.919, all P<0.01). The results of pathway cluster analysis showed that in the comparison of differential proteins between IC group and NC group, only the platelet degranulation pathway was enriched, and 10 proteins were involved in this pathway (P<0.01). In the comparison of differential proteins between UC group and NC group, there were 58 pathways enriched, of which 38 proteins were involved in the platelet degranulation pathway, 16 proteins were involved in the initial complement trigger pathway, 13 proteins were involved in the complement cascade pathway, and 11 proteins were involved in antibody-mediated complement activation pathway (all P<0.01). In the comparison of differential proteins between UC group and IC group, three different pathways were obtained. Among them, nine proteins were involved in the platelet degranulation pathway, seven proteins were involved in the initial complement trigger pathway, and five proteins were involved in the complement cascade pathway (all P<0.01). Conclusions The difference in serum proteome between IC patients and UC patients was significant, and the differential proteins are mainly involved in platelet activation and complement activation. The candidate proteins identified in this study may be used as biomarkers for the differential diagnosis of UC and IC in the future.