[Correlation analysis of clock genes and MEN2 medullary thyroid carcinoma].

Objective: To investigate the correlation between CLOCK and BMAL1 genes and MEN2 medullary thyroid carcinoma (MTC). Methods: Thirteen cases with MEN2 MTC and thirteen cases with non-MEN2 MTC were selected who were treated in the Yantai Yuhuangding Hospital between January 2013 and September 2021. Clinical indicators such as blood calcitonin level, tumor diameter and metastatic lymph node of patients were collected. The expression differences of CLOCK and BMAL1 between MEN2 MTC and para-carcinoma tissue as well as between MEN2 MTC and non-MEN2 MTC were detected by immunohistochemistry and qPCR. The correlation between lymph node metastasis and CLOCK or BMAL1 expression was analyzed. Protein-protein interaction (PPI) network analysis combined with qPCR and correlation analysis was used to explore the expression regulation relationship between RET and circadian clock genes. The rhythm disorder of MEN2 cells was verified by lipopolysaccharide cell stimulation experiment after dexamethasone rhythm synchronization. Results: MEN2 MTC exhibited typical RET gene mutation. The mean blood calcitonin level, the tumor diameter and the number of metastatic lymph nodes of patients with MEN2 MTC were higher than those of patients with non-MEN2 MTC (t value was 2.76, 2.53, 2.26, all P<0.05). Immunohistochemical results showed that the expression levels of CLOCK and BMAL1 in MEN2 MTC were higher than those in non-MEN2 MTC, while negatively expressed in para-cancerous thyroid follicle. qPCR displayed that the expression of CLOCK gene in cancer tissues was higher than that in non-MEN2 MTC and para-cancerous tissues (t value was 2.68 and 2.86, all P<0.05); the expression of BMAL1 gene in MEN2 MTC was higher than that in non-MEN2 MTC and para-cancerous tissues (t value was 2.21 and 2.35, all P<0.05). Correlation analysis showed that the expression levels of CLOCK and BMAL1 genes were positively correlated with the number of lymph node metastases in patients with MEN2 MTC (r=0.65, P<0.001; r=0.52, P=0.005). PPI network analysis indicated that the expression of CLOCK gene was positively correlated with the abnormal expression of RET gene (r=0.96, P<0.001). With lipopolysaccharide to stimulate cultured cells in vitro after dexamethasone rhythm synchronization, the expressions of CLOCK and BMAL1 in MEN2 MTC cells (0.47±0.22 and 2.60±1.48) at 12 hours of synchronization were significantly lower than those in para-cancerous tissues (1.70±1.62 and 8.23±2.52), the difference was statistically significant(t=5.04, P=0.007; t=3.34, P=0.029). Conclusion: CLOCK and BMAL1 are correlated with the occurrence and development of MEN2 MTC, and may be potential targets for the development of new therapeutic strategies for MEN2 MTC.

Hypereosinophilic syndrome presenting with bilateral ear fullness.

INTRODUCTION: Hypereosinophilic syndrome is a rare disease with hypereosinophilia resulting in end-organ dysfunction. Patients present with organ-associated symptoms, and the targets frequently affected are heart, lung, skin, or the nervous system, and the middle ear involvement is rare. CASE REPORT: A 30-year-old female with left ear fullness and hearing loss, which persisted for 6months, was finally diagnosed with hypereosinophilic syndrome (HES). After high dose systemic steroids treatment, all symptoms improved. CONCLUSION: Eosinophilic otitis media and HES involving the middle ear share many clinical manifestations. Prompt and accurate differential diagnosis is required for these diseases to ameliorate symptoms and promote recovery.

Decreased expression of ferritin light chain in osteosarcoma and its correlation with epithelial-mesenchymal transition.

OBJECTIVE: The purpose of this study was to detect ferritin light chain (FTL) expression level in osteosarcoma (OS), and to clarify whether FTL could offer additional help in diagnosis or therapy. MATERIALS AND METHODS: First, we assessed FTL level in OS tissues and cells through GEO dataset and tissue microarrays (TMAs). Then, we overexpressed FTL expression in MG-63 cell line. Lastly, we detected the expression of EMT-related signal pathway proteins to study its underlying molecular mechanisms. RESULTS: GEO dataset and TMAs showed that FTL was down-regulated in OS. After FTL was overexpressed, the proliferation, migration and invasion abilities of OS cells were significantly reduced. Moreover, after FTL overexpressing, the levels of CDH2 and Vimentin were down-regulated with CDH1 up-regulated. CONCLUSIONS: We revealed that FTL (1) is lower in OS then in normal tissue, (2) is related to metastasis, survival period, and therapeutic response, and (3) may be a tumor-inhibiting factor owing to its inhibition of EMT in OS.

Preset ureter catheter in laparoscopic radical hysterectomy of cervical cancer.

The aim of this study was to evaluate the clinical efficacy of a temporary ureteral catheter in preventing iatrogenic ureteral damage in cervical cancer patients undergoing laparoscopic radical hysterectomy. All cases had confirmed diagnoses of cervical cancer preoperatively between December 2008 and December 2012 in our hospital and were in clinical stages IA2 to IIA. In total, 176 laparoscopic radical hysterectomy and lymphadenectomy procedures were performed. The 176 cases were divided into two groups: ureteral catheters were installed using cystoscopy before the operation in 86 patients (group A), and ureteral catheters were not placed in 90 patients (group B). These cases were retrospectively analyzed based on postoperative hospitalization time and intraoperative and postoperative complications. A total of 6 cases (3.41%) had ureteral injuries, and 4 of the cases (4.65%) of ureteral injuries occurred in group A. In two of these cases, urinary leaking appeared at the post-operative 8th and 9th days and at the 10th and 25th days, respectively. There were 2 cases (2.22%) of ureteral injuries in group B: 1 case of intraoperative direct injury and the other of urinary leaking, which appeared at post-operative day 21. Statistically significant differences between the two groups were observed in operating time and the incidence of hemorrhage, hematuria (including microscopic hematuria), post-operative urinary tract infection, and pain (P < 0.05). A ureteral catheter that is placed preoperatively can help to identify the ureter in laparoscopic radical hysterectomy, but does not decrease the incidence of ureteral injury.

Left atrial enlargement induced by pure mitral regurgitation: time frame in a new swine model.

BACKGROUND: Atrial enlargement occurs in patients with significant mitral regurgitation. However, the time frame of the development of atrial enlargement induced by mitral regurgitation remains unknown. METHODS: Fourteen Lanyu miniature pigs (age = 6.6 ± 0.9 months) were studied. Mitral regurgitation was created by placing a predefined hole on the middle scallop of the posterior mitral leaflet under cardiopulmonary bypass. The parasternal long-axis atrial dimension was measured by transthoracic echocardiographic examinations. RESULTS: All animals exhibited grade 3 mitral regurgitation immediately after surgery. Seven pigs expired within 2 weeks after the operation [technical complications (n = 1), acute cardiac tamponade (n = 1), and acute and subacute heart failure (n = 5)]. Seven pigs remained alive at a mean follow-up of 7.7 ± 2.1 months. The left atrial diameter indices of the 7 pigs increased significantly at 1 month (33.1 ± 8.6 mm, p = 0.018) and 3 months (41.3 ± 12.6 mm, p = 0.018) after surgery compared with baseline values (22.8 ± 5.2 mm), and the left atrial diameter index increased significantly at 3 months compared to 1 month (p = 0.018). CONCLUSIONS: Left atrial enlargement develops rapidly and progresses after the creation of significant pure mitral regurgitation.

Blends of stearyl poly(ethylene oxide) coupling-polymer in chitosan as coating materials for polyurethane intravascular catheters.

To optimize the surface biocompatibility of the intravascular catheter, an amphiphilic coupling-polymer of stearyl poly (ethylene oxide) -co- 4,4'-methylene diphenyl diisocyanate-co- stearyl poly (ethylene oxide), for short MSPEO, was specially designed as the surface modifying additive (SMA). The blend of MSPEO in chitosan was coated on the outer wall of the catheters by the dip-coating method. The surface analysis was carried out by ATR-FTIR and contact angle measurements. The surface enrichment of MSPEO was confirmed. On the water interface, the larger the molecular weight of PEO was, the higher the surface enrichment. While on air interface, the case was the contrary. Three kinds of static test of clotting time, plasma recalcification time (PRT), prothrombin time (PT), and thrombin time (TT), as well as the static platelet adhesion experiment were carried out. The results indicated that the coated surface could resist the clotting effectively. In order to test the blood-compatibility of the coated catheters under a shear of blood flow, the dynamic experiment was performed through a closed-loop tubular system with the shear rate of 1500 s(-1). The results of blood regular testing at six different times (0, 5,10, 20, 30, and 60 min) indicated that the biocompatibility of the coating was nearly ideal. Finally, the SMA-MSPEO was proved to be non-chronic-toxic by animal experiments with rats and suitable as a coating material for clinical use.

Surface coating of stearyl poly(ethylene oxide) coupling-polymer on polyurethane guiding catheters with poly(ether urethane) film-building additive for biomedical applications.

Three types of stearyl poly(ethylene oxide) (SPEO) with Mn of 2,300, 6,000 and 12,000 were synthesized; accordingly, three types of amphiphilic coupling-polymer SPEO-MDI-SPEO (MSPEO) were prepared by the reactions with 4,4'-methylene diphenyl diisocyanate (MDI). As the surface-modifying additives (SMA), MSPEOs were coated onto the outer wall of the medical guiding catheters. Due to the lack of stability, when coated, MSPEO blended with the film building agent (FBA), poly(ether urethane) (PEL). The process of coating was performed with a lifter. With invariable speed, the PU guiding catheter was vertically dipped into the coating mixture of SMA-MSPEO and FBA-PEL. The surface analysis was carried out by ATR-FTIR and contact angle measurements. It was proved that the surface enrichment of PEO on water interface was much higher than that on air interface. Three kinds of static clotting time tests, PRT, PT and TT, as well as the static platelet adhesion experiment were performed. The results indicated that the coated surface could resist the blood coagulation effectively. In order to test the blood compatibility of the coated catheters under a shear of blood flow, the dynamic experiment was performed with a closed-loop tubular system under a shear rate of 1,500 s(-1). The blood regular testing was carried out on the samples taken out at six different times (0, 5,10, 20, 30 and 60 min). The results were ideal. Finally, the SMA-MSPEO was proved to be non-acute-toxic by LD50 test.

Changing clinical course of patients with malignant mesothelioma: implications for FNA cytology and utility of immunocytochemical staining.

The diagnosis of both local recurrences and distant metastases of mesothelioma can be accomplished by fine-needle aspiration (FNA) biopsy. Although the previous history of mesothelioma provides strong support for recurrent/metastatic mesothelioma, other diagnostic possibilities (particularly adenocarcinoma) may require exclusion via special stains in some cases. In this study, we report on the morphologic findings in 13 cases of mesothelioma which underwent FNA (7 metastatic lesions, 6 local recurrences). In addition, immunohistochemical staining results for 7 cases with available material using antibodies directed against cytokeratin AE 1/3 and two antibodies reported to show consistently positive results in mesothelioma (calretinin and cytokeratin 5/6) are reported and compared to results seen for 10 cases of adenocarcinoma. All cases of mesothelioma and adenocarcinoma showed strong staining with cytokeratin AE 1/3. Three of 7 cases of mesothelioma showed strong staining with calretinin, while only focal staining was detected in 3 additional cases; only one case showed positive staining with cytokeratin 5/6. One of 10 cases of adenocarcinoma showed calretinin positivity; however, at least focal staining with cytokeratin 5/6 was seen in 4 cases. These results suggest that cytokeratin 5/6 is neither a sensitive nor specific stain for the diagnosis of mesothelioma in cytology material. Calretinin appears to be more specific for mesothelioma but showed disappointing sensitivity for this tumor, potentially limiting its diagnostic utility in FNA material.

Adenocarcinoma in colonic brushing cytology: High-grade dysplasia as a diagnostic pitfall.

Cytologic evaluation of brushing specimens obtained from the colon may be useful in the diagnosis of neoplastic and inflammatory lesions, as previous studies have reported favorable sensitivity and specificity figures for this procedure. In this study, we report our experience with 80 colonic brushings examined over a 5-yr period. Thirty cases received an atypical or malignant cytologic diagnosis. Nineteen of 20 cases diagnosed cytologically as adenocarcinoma revealed adenocarcinoma on biopsy; one case showed only adenomatous epithelium on biopsy and subsequent resection. Cases diagnosed cytologically as "atypical" or "adenomatous" showed adenocarcinoma, adenoma, and inflammatory conditions upon biopsy. Slides from 30 atypical/malignant cases were retrospectively reviewed for a number of cytomorphologic features and were correlated with the histologic diagnosis. Cases from histologically confirmed adenocarcinoma tended to show greater degrees of altered nuclear polarity, nuclear pleomorphism, membrane irregularities, and chromatin pattern alterations than those from histologically proven adenomatous or inflammatory lesions. The most likely cause of a false-positive diagnosis in this setting is sampling of an adenoma with high-grade dysplasia which fails to meet histologic criteria for adenocarcinoma (invasion of the underlying muscularis mucosae). Thus, in the second part of the study, we examined histologic sections from surgically excised adenomas to determine the frequency with which profound nuclear atypia is at least focally present, potentially resulting in a false-positive cytology diagnosis upon brushing. Slides from 51 cases were reviewed; cytologic atypia beyond that typically observed in adenomas was not observed in 43% of cases. However, profound nuclear atypia was present in 6% of cases; cytologic evaluation of a brushing specimen from these lesions may have resulted in a false-positive diagnosis of adenocarcinoma, despite the histologic diagnosis of adenoma with severe dysplasia. The remaining cases demonstrated intermediate degrees of atypia. These findings serve to quantitate the frequency with which cytohistologic discrepancies might be expected for mass lesions of the colon.

Evaluation of mild-to-moderate dysplasia on cervical-endocervical (Pap) smear: a subgroup of patients who bridge LSIL and HSIL.

The Bethesda System recommends Pap smear diagnosis to be based on the most abnormal cells present, regardless of number. Our reporting system includes a diagnostic category of mild-to-moderate dysplasia (D1-2), defined as cases with only a few moderately dysplastic cells. We evaluated the validity of a D1-2 diagnostic category by reevaluation of 58 cases with subsequent follow-up (up to 24 months). On biopsy and/or Pap smear follow-up, 24 cases (41%) showed LSIL and 34 cases (59%) showed HSIL. Index smears from these cases were examined by two cytopathologists blinded to patient follow-up for the following morphologic features: volumes (scale 1-4) of LSIL, HSIL, and dyskeratosis, HSIL as single cells or syncytial fragments, and acute inflammation. None of the morphologic features evaluated were significantly different between the LSIL and HSIL follow-up groups based on univariate and multivariate logistic regression analysis. The diagnosis of D1-2 on Pap smear is a valid diagnostic category that defines a subgroup of patients with both LSIL and HSIL follow-up, which cannot be reliably predicted based on morphology alone.

Follow-up of morphologically reactive lymphoid proliferations in fine-needle aspirates of elderly patients.

Fine-needle aspiration (FNA) is an effective tool in evaluating the cause of lymphadenopathy. While the morphologic diagnosis of a reactive lymphoid proliferation is common in younger patients, this diagnosis should be made carefully in older patients (those over the age of 50 yr) in light of the facts that such purely reactive conditions occur much less frequently in this population, and that follow-up of these patients reveals a malignancy (usually lymphoma) in a significant number of cases. In this series, we identified 40 patients with a morphologic diagnosis of reactive lymphoid proliferation on FNA and obtained their follow-up information. Of 19 patients under the age of 50 yr, 5 underwent subsequent biopsies and only one revealed a definitive malignancy (5%). In contrast, 7 of 21 patients over the age of 50 yr underwent a subsequent biopsy, and 6 were found to have a malignancy (5 malignant lymphomas, 1 metastatic melanoma). The higher rate of positive follow-up (29%) in this age group supports previous suggestions that morphologically reactive (mixed) lymphoid proliferations be viewed with increased suspicion in the elderly patient, and that additional studies, such as flow cytometry, be performed when material is available.

Intercellular adhesion molecule expression in ductal carcinoma of the breast: correlation of immunohistochemical staining with cytologic smear pattern.

Recent studies suggest that altered expression of intercellular adhesion molecules (ICAM) in ductal carcinoma of the breast is associated with a higher incidence of metastases and decreased patient survival. In addition, the presence of significant cellular dyscohesion in cytologic smear preparations has been found to correlate with the presence of regional and distant metastases in a subset of patients. In this study, we correlate the smear pattern in preparations taken directly from surgically excised breast tumors with their immunohistochemical staining pattern, using antibodies directed against a panel of ICAM. We found excellent correlation, as all three tumors with an extremely high degree of tumor cell cohesion showed strong staining with all ICAM antibodies in the vast majority (>/=90%) of tumor cells in corresponding tissue sections. In contrast, five tumors displaying a largely dyscohesive smear pattern demonstrated decreased staining (

Fine-needle aspiration of cystic parotid glands lesions: an institutional review of 46 cases with histologic correlation.

BACKGROUND: Many case reports describing various benign and malignant disease entities diagnosed in cystic parotid gland lesions by fine-needle aspiration (FNA) exist in the literature. Very few studies, however, discuss the efficacy and the diagnostic difficulties of FNA cytology in such lesions. The authors report a 5-year institutional experience with FNA cytology of cystic parotid gland lesions and address the accuracy of the procedure and avoidance of diagnostic pitfalls. METHODS: A retrospective review of 46 cases in which patients clinically presented with cystic parotid gland lesions was done from a total of 221 parotid FNAs performed over a 5-year period at the University of Pennsylvania Medical Center. The clinical features, cytology, and follow-up surgical pathology were reviewed to determine diagnostic accuracy and highlight potential pitfalls. RESULTS: Based on subsequent surgical excision in 29 of the 46 cases of cystic parotid gland lesions, a diagnostic accuracy rate of 83% was obtained for FNA. Approximately 20% (9 of 46 cases) were found to be clinically significant tumors and included 4 mucoepidermoid and 1 adenocarcinoma (NOS), 3 benign mixed tumors, and 1 metastatic melanoma. Three major factors were implicated as diagnostic pitfalls in our series: 1) failure to obtain critical clinical information, 2) overinterpretation of paucicellular specimens, and 3) failure to realize that certain tumors generally perceived as solid masses can present clinically as cystic lesions. CONCLUSIONS: FNA cytology is a valuable tool in the primary diagnosis and management of cystic parotid gland lesions. The diagnostic accuracy of this procedure can be significantly improved by acquiring a detailed clinical history, obtaining an adequate cellular specimen, and having knowledge of the variety and frequencies of possible diagnostic entities that may present as cystic parotid gland lesions.