Objective:To analyze the relationship between tumor-associated immune cells and cytokines with papillary thyroid carcinoma (PTC) and explore the correlation between tumor-associated immune cells and the risk stratification of PTC recurrence.Methods:A total of 58 PTC patients diagnosed by surgical pathology from Aug. 2022 to Aug. 2023 in the General Surgery and Thyroid and Hernia Department of Tianjin Medical University General Hospital were selected. All patients underwent thyroidectomy combined with central or lateral cervical lymph node dissection. According to lymph node metastasis status, patients were divided into lymph node metastasis-negative (22 cases) and lymph node metastasis-positive groups (36 cases) based on post-operative pathological diagnosis. Flow cytometry was used to detect the levels of tumor-associated immune cells (T, B, NK cells) and cytokines (IL-10, IL-17, IL-35, IFN-γ) in T cells.Results:Compared to normal tissue located distant from the cancer, a significant increase in the proportion of NK cells was observed in cancerous tissue ( t=-2.11, P=0.032). Similarly, the proportion of CD8+ T cells was also significantly elevated ( t=-5.12, P=0.005). In lymph node tissue, the proportion of CD4+ T cells in LNM-positive tissue was significantly higher than in LNM-negative tissue ( t=-3.89, P=0.004), while the proportion of CD8+ T cells exhibited a significant decrease ( t=2.41, P=0.004). Additionally, the levels of IL-10 in cancerous tissue were significantly elevated ( t=-3.83, P=0.003), as were the levels of IL-17 ( t=-4.83, P=0.003). In lymph node tissues categorized by LNM status, although not statistically significant, the levels of IL-10 and IL-17 were generally higher in LNM-positive cases compared to LNM-negative cases. Among the 58 cases stratified by recurrence risk, 22 cases (37.9%) were classified as low-risk, while 36 cases (62.1%) were classified as intermediate-risk. The differences in the proportions of CD4+ T and CD8+ T cell subsets in the lymph nodes of PTC patients were statistically significant. Logistic regression analysis indicated that a higher proportion of CD4+ T cell subsets in the lymph nodes of PTC patients was associated with a higher recurrence risk stratification compared to those with a lower proportion ( OR=1.107, 95% CI: 1.001-1.224). Using the predicted probability as the test variable and "low and medium risk" as the state variable, a ROC curve was constructed, yielding an AUC of 0.790 with P=0.003, indicating a good predictive effect of the model on the dependent variable. Conclusions:Compared to normal tissues located distant from cancerous regions, cancer tissues exhibit a significantly elevated proportion of tumor-associated immune cells and cytokine levels, thereby creating an immunosuppressive tumor microenvironment. Additionally, patients with papillary thyroid carcinoma (PTC) who are classified as having a higher risk of recurrence demonstrate a greater proportion of CD4+ T cell subsets in their lymph nodes.

Hyperparathyroidism-jaw tumor syndrome (HPT-JT) is a rare familial autosomal dominant genetic disease with primary hyperparathyroidism, jaw tumors, kidney tumors and uterine tumors caused by cell division cycle 73 (CDC73) germline mutations. A 42-year-old male patient was admitted for pancreatitis and further examination revealed elevated PTH at 54.00pmol/L and a history of jaw tumors. This patient was diagnosed as HPT-JT finally and underwent upper right, lower right, and upper left parathyroid glands resection and genetic testing. Postoperative pathology revealed that atypical adenomatous nodules of parathyroid glands with extensive atypia and nucleus division and parathyroid hyperplasia and whole exome sequencing suggested that the CDC73 mutation.

Objective To construct a Ca2+/calmodulin-dependent protein kinase Ⅱ(CaMKⅡ)long-fragment fusion protein plasmid;investigate the expression,extraction,and purification of CaMK Ⅱ;and identify its binding to the Cav1.2 channel.Methods The extracted pGEX-6p-1/CaMK Ⅱ long-fragment plasmid was transformed into Escherichia coli BL21 receptor cells and cultured in a shaking incubator for 12 h.Isopropyl β-D-thiogalactoside was added to promote GST fusion protein expression.Next,the GST-CaMK Ⅱ long frag-ment was isolated and purified with GS-4B using dithiothreitol(DTT)combined with ultrasonic crushing.After treatment with the PreScis-sion protease,the GST label was removed to obtain the CaMK Ⅱ long-fragment protein.The molecular weight and relative purity of the CaMKⅡ long-fragment protein were determined using 15％sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE).The concentration of the purified protein was determined using the Bradford method.The binding ability of the CaMK Ⅱ long-fragment pro-tein to the Cav1.2 channel protein was evaluated using the pull-down method combined with Western blotting.Results The sequencing results showed that the CaMK Ⅱ long fragment was successfully constructed.A CaMK Ⅱ long-fragment protein with high purity and con-centration was obtained using DTT combined with ultrasonic crushing.This protein can bind to the CT1 protein of cardiac Cav1.2 calcium channel.Conclusion In this study,we successfully constructed a CaMKⅡ long-fragment plasmid.The CaMKⅡ long-fragment protein was extracted and purified,and was determined to bind to Cav1.2 channel proteins and exhibit biological activity.Collectively,this study provides a basis for further study of the function of CaMK Ⅱ.

Objective:To analyze the relationship between tumor-associated immune cells and cytokines with papillary thyroid carcinoma (PTC) and explore the correlation between tumor-associated immune cells and the risk stratification of PTC recurrence.Methods:A total of 58 PTC patients diagnosed by surgical pathology from Aug. 2022 to Aug. 2023 in the General Surgery and Thyroid and Hernia Department of Tianjin Medical University General Hospital were selected. All patients underwent thyroidectomy combined with central or lateral cervical lymph node dissection. According to lymph node metastasis status, patients were divided into lymph node metastasis-negative (22 cases) and lymph node metastasis-positive groups (36 cases) based on post-operative pathological diagnosis. Flow cytometry was used to detect the levels of tumor-associated immune cells (T, B, NK cells) and cytokines (IL-10, IL-17, IL-35, IFN-γ) in T cells.Results:Compared to normal tissue located distant from the cancer, a significant increase in the proportion of NK cells was observed in cancerous tissue ( t=-2.11, P=0.032). Similarly, the proportion of CD8+ T cells was also significantly elevated ( t=-5.12, P=0.005). In lymph node tissue, the proportion of CD4+ T cells in LNM-positive tissue was significantly higher than in LNM-negative tissue ( t=-3.89, P=0.004), while the proportion of CD8+ T cells exhibited a significant decrease ( t=2.41, P=0.004). Additionally, the levels of IL-10 in cancerous tissue were significantly elevated ( t=-3.83, P=0.003), as were the levels of IL-17 ( t=-4.83, P=0.003). In lymph node tissues categorized by LNM status, although not statistically significant, the levels of IL-10 and IL-17 were generally higher in LNM-positive cases compared to LNM-negative cases. Among the 58 cases stratified by recurrence risk, 22 cases (37.9%) were classified as low-risk, while 36 cases (62.1%) were classified as intermediate-risk. The differences in the proportions of CD4+ T and CD8+ T cell subsets in the lymph nodes of PTC patients were statistically significant. Logistic regression analysis indicated that a higher proportion of CD4+ T cell subsets in the lymph nodes of PTC patients was associated with a higher recurrence risk stratification compared to those with a lower proportion ( OR=1.107, 95% CI: 1.001-1.224). Using the predicted probability as the test variable and "low and medium risk" as the state variable, a ROC curve was constructed, yielding an AUC of 0.790 with P=0.003, indicating a good predictive effect of the model on the dependent variable. Conclusions:Compared to normal tissues located distant from cancerous regions, cancer tissues exhibit a significantly elevated proportion of tumor-associated immune cells and cytokine levels, thereby creating an immunosuppressive tumor microenvironment. Additionally, patients with papillary thyroid carcinoma (PTC) who are classified as having a higher risk of recurrence demonstrate a greater proportion of CD4+ T cell subsets in their lymph nodes.

Hyperparathyroidism-jaw tumor syndrome (HPT-JT) is a rare familial autosomal dominant genetic disease with primary hyperparathyroidism, jaw tumors, kidney tumors and uterine tumors caused by cell division cycle 73 (CDC73) germline mutations. A 42-year-old male patient was admitted for pancreatitis and further examination revealed elevated PTH at 54.00pmol/L and a history of jaw tumors. This patient was diagnosed as HPT-JT finally and underwent upper right, lower right, and upper left parathyroid glands resection and genetic testing. Postoperative pathology revealed that atypical adenomatous nodules of parathyroid glands with extensive atypia and nucleus division and parathyroid hyperplasia and whole exome sequencing suggested that the CDC73 mutation.

Objective To construct a Ca2+/calmodulin-dependent protein kinase Ⅱ(CaMKⅡ)long-fragment fusion protein plasmid;investigate the expression,extraction,and purification of CaMK Ⅱ;and identify its binding to the Cav1.2 channel.Methods The extracted pGEX-6p-1/CaMK Ⅱ long-fragment plasmid was transformed into Escherichia coli BL21 receptor cells and cultured in a shaking incubator for 12 h.Isopropyl β-D-thiogalactoside was added to promote GST fusion protein expression.Next,the GST-CaMK Ⅱ long frag-ment was isolated and purified with GS-4B using dithiothreitol(DTT)combined with ultrasonic crushing.After treatment with the PreScis-sion protease,the GST label was removed to obtain the CaMK Ⅱ long-fragment protein.The molecular weight and relative purity of the CaMKⅡ long-fragment protein were determined using 15％sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE).The concentration of the purified protein was determined using the Bradford method.The binding ability of the CaMK Ⅱ long-fragment pro-tein to the Cav1.2 channel protein was evaluated using the pull-down method combined with Western blotting.Results The sequencing results showed that the CaMK Ⅱ long fragment was successfully constructed.A CaMK Ⅱ long-fragment protein with high purity and con-centration was obtained using DTT combined with ultrasonic crushing.This protein can bind to the CT1 protein of cardiac Cav1.2 calcium channel.Conclusion In this study,we successfully constructed a CaMKⅡ long-fragment plasmid.The CaMKⅡ long-fragment protein was extracted and purified,and was determined to bind to Cav1.2 channel proteins and exhibit biological activity.Collectively,this study provides a basis for further study of the function of CaMK Ⅱ.

Humans ; Parathyroid Neoplasms/genetics* ; Molecular Biology ; Parathyroid Glands

Objective:To investigate the incidence characteristics and influencing factors of non-alcoholic fatty liver disease (NAFLD) in rural Uyghur ethnic group residents in Xinjiang Production and Construction Corps and to provide scientific evidence for early identification and prevention of NAFLD for residents.Methods:A total of 10 158 participants were included from the Xinjiang Uygur ethnic group population cohort. A prospective cohort study and Cox proportional hazards regression model analysis were used to explore the influencing factors and clustering of NAFLD, and the dose-response relationship between related biochemical indicators and the risk of NAFLD was studied using a restricted cubic spline.Results:The cumulative incidence rate of NAFLD was 6.9%, and the incidence density of NAFLD was 12.06/1 000 person-years. The incidence density of NAFLD in females was higher than in males (14.72/1 000 person-years vs. 9.17/1 000 person-years, P<0.001). The incidence density of NAFLD gradually increased with age in the total population, both men and women (all P<0.001). In the general population, an education level of junior high school or above was a protective factor for NAFLD, while older age, divorce, widowhood, overweight, obesity, hypertension, increased glomerular filtration rate, decreased HDL-C, increased LDL-C, and increased ALT were risk factors for NAFLD. Estimated glomerular filtration rate (eGFR), HDL-C, LDL-C, and ALT were non-linearly correlated with the incidence of NAFLD, and there was a significant dose-response relationship between them. Only 19.1% of residents had no NAFLD risk factors; over 80.9% had ≥1 NAFLD risk factors. The risk of NAFLD increased with the number of risk factors. Conclusions:The incidence of NAFLD in rural Uygur ethnic group residents in Xinjiang Production and Construction Corps was relatively low, but most residents had one or more risk factors for NAFLD. Prevention and control of NAFLD in this population cannot be ignored. In addition, people of older age, divorced or widowed, low education level, overweight or obese, hypertension, and abnormal eGFR, HDL-C, LDL-C, and ALT were the high-risk groups of NAFLD that need to be paid attention to in this population.

Currently, the main treatments for nonfunctional parathyroid cysts (NFPC) are fluid aspiration,sclerosing injection and surgical removal. The choice of treatment method is controversial. Eight patients with NFPC who were treated by simple aspiration combined with parathyroid hormone (PTH) rapid determination in General Surgery Department of Tianjin Medical University General Hospital from Dec. 2020 to Oct. 2022 are reported to provide a reference for the choice of treatment, which can also reduce surgical pain and accidental sclerosing injury.

Objective:To investigate the clinical features, diagnostic methods, treatment modalities, and prognosis of parathyroid multiglandular disease (PTMGD) in primary hyperparathyroidism (PHPT) .Methods:The clinical data of 29 patients with PTMGD who underwent surgery at the General Surgery Department of the Tianjin Medical University General Hospital from Dec. 2015 to Jul. 2023 were retrospectively analyzed, including the patients' preoperative and postoperative blood calcium and parathyroid hormone, the main clinical manifestations, the involvement of other systems, the main types of pathology, the accuracy of the various examinations, and the postoperative prognosis, etc., and were compared with the 291 patients who had undergone surgery for single-glandular lesion patients were compared. SPSS25 was used to analyze the data.Results:The age of onset of PTMGD was 52.7±1.9 years compared to 56.6±0.7 years in patients with monoglandular disease, P=0.047. Tumor diameter of PTMGD was (2.05±0.1) cm and (2.34±0.6) cm of monoglandular disease, P=0.006. The preoperative blood calcium was (2.56±0.59) mmol/L in PTMGD and (2.70±0.58) mmol/L in monoglandular disease, P=0.045. Preoperative parathyroid hormone (PTH), blood calcium, and Win values were positively correlated with maximum tumor diameter in patients with PTMGD (R-values of 0.362, 0.223, and 0.352, respectively) .Neck ultrasound, neck-enhanced CT and parathyroid nuclear imaging were used to localize and diagnose the diseased parathyroid glands in this group of cases.The accuracy rates were (14/25) 56%, (10/19) 53% and (11/24) 46% in patients with PTMGD, while in patients with monoglandular disease, the accuracy rates were (233/250) 89%, (131/152) 96% and (223/232) 86%. PTMGD accuracy rate was less than that of monoglandular disease,and was statistically significant ( P-value was less than 0.001 in all cases) .The accuracy of the combined localization diagnosis of the three tests in patients with PTMGD was then improved to (13/18) 72%. The pathology of PTMGD was predominantly parathyroid hyperplasia, 45/72 (63%), compared to that of monoadenopathy 18/291 (6%), P<0.001. Parathyroid adenomas predominated in patients with monoadenopathy compared to that of PTMGD, 237/291 (82%) vs. 24/72 (33%), and the proportion of parathyroid adenomas in patients with monoadenopathy was higher than that in patients with PTMGD, P<0.001. 23 patients with PTMGD were followed up, of whom 9 showed mild elevation of parathyroid hormone postoperatively, and 1 patient showed signs of hypoparathyroidism. Conclusion:The low age of onset of multiglandular lesions in primary hyperparathyroidism, mild biochemical tests, and the difficulty of accurately locating all lesions preoperatively warrant adequate preoperative evaluation to promptly identify patients with familial multiple endocrine adenomas, as well as intraoperative bilateral parathyroid exploration in patients with suspected multiglandular lesions.