Molecular pathology assists the diagnosis of lymphoepithelial sialadenitis, Sjögren's syndrome and extranodal marginal zone lymphoma of mucosa-associated lymphoid tissue.

Background/purpose: Lymphoepithelial sialadenitis (LESA), Sjögren's syndrome (SS), and salivary MALT lymphoma are diseases characterized by lymphoepithelial lesions, and the differential diagnosis between them in the salivary glands is challenging. This study aimed to explore clinicopathological and genetic characteristics of the three diseases. Materials and methods: We retrospectively analyzed the clinical features, the histomorphology, immunohistochemistry, and genetic profiling by polymerase chain reaction (PCR) and next-generation sequencing (NGS). Results: There included 68 LESAs, 25 SSs, and 62 MALT lymphomas. Ten cases relapsed in total, and 3 of MALT lymphomas died due to high-level transformation. Immunohistochemical double staining showed FCRL4 cells co-expressed Pax-5 and Ki-67, suggesting FCRL4 cells were proliferative B-cells. The expression level of the FCRL4 was significantly higher in MALT lymphoma than LESA and SS. The detection rates of clonal IGH, IGK, and IGL gene rearrangements in MALT lymphoma with a sensitivity of 83.33%. Monoclonal immunoglobulin gene rearrangements were confirmed in five suspected patients by NGS (100%). Conclusion: FCRL4 B cells might play an important role in the formation of lymphoepithelial lesions and might be as a diagnostic positive marker of salivary MALT lymphoma. The application of multiple detection methods could significantly improve the diagnostic accuracy for MALT lymphomas from LESA and SS.

Conservative rehabilitation therapy for respiratory dysfunction due to phrenic nerve sacrifice during resection of massive mediastinal tumor: A case series study.

RATIONALE: Cases of respiratory dysfunction due to phrenic nerve sacrifice during resection of massive mediastinal tumor have rarely been studied in detail. Diaphragmatic dysfunction in such cases can lead to potentially fatal respiratory and circulatory disturbances. Therefore, timely diagnosis and intervention are important. Conservative rehabilitation therapy is the first choice for respiratory dysfunction due to diaphragmatic dysfunction. PATIENT CONCERNS, DIAGNOSES AND INTERVENTIONS: We present 3 patients with respiratory dysfunction due to phrenic nerve sacrifice during resection of massive mediastinal tumor. The diagnostic methods and therapeutic procedures for diaphragmatic dysfunction for each patient are described in detail. This study highlights the role of ventilator support combined with physical therapy in the treatment of respiratory dysfunction in such cases. The diagnosis of diaphragmatic dysfunction as well as the risk assessment of phrenic nerve involvement are also discussed. The modalities of ventilator support, including modes and parameters, are listed. OUTCOMES AND LESSONS: This study provides experiences of diagnosis and treatment of respiratory dysfunction due to phrenic nerve sacrifice during resection of massive mediastinal tumor. Timely diagnosis of diaphragmatic dysfunction primarily relies on clinical manifestations and radiography. Conservative rehabilitation therapy can improve or restore diaphragmatic function in majority of patients, and avert or delay the need for surgical intervention. Preoperative assessment of the risk of phrenic nerve involvement is important in such cases.

Expression and significance of m6A-RNA-methylation in oral cancer and precancerous lesion.

Background: Oral potentially malignant disorders (OPMDs) and oral squamous cell carcinoma (OSCC) are a series of related pathologic and molecular events involving simple epithelial hyperplasia, mild to severe dysplasia and canceration. N6-methyladenosine RNA methylation, as the most common modification of both coding mRNA and non-coding ncRNA in eukaryotes, participates in the regulation of the occurrence and development of various malignant tumors in human. However, its role in oral epithelial dysplasia (OED) and OSCC remain unclear. Materials and methods: In this study, multiple public databases were used for bioinformatics analysis of 23 common m6A methylation regulators in head and neck squamous cell carcinoma (HNSCC). Protein expressions of IGF2BP2 and IGF2BP3 were verified accordingly in clinical cohort samples of OED and OSCC. Results: Patients with high expression of FTO、HNRNPC、HNRNPA2B1、LRPPRC、IGF2BP1、IGF2BP2、IGF2BP3 had a poor prognosis. IGF2BP2 had a relatively high mutation rate in HNSCC, and its expression was significantly positively correlated with tumor purity, and significantly negatively correlated with the infiltration level of B cells and CD8+T cells. The expression of IGF2BP3 was significantly positively correlated with tumor purity and CD4+T cells. Immunohistochemistrically, the expression of IGF2BP2 and IGF2BP3 in oral simple epithelial hyperplasia, OED and OSCC increased gradually. Both were strongly expressed in OSCC. Conclusion: IGF2BP2 and IGF2BP3 were the potential biological prognostic indicators of OED and OSCC.

[Preliminary establishment of an unsupervised quantification model of Ki-67 proliferation index based on local variable threshold method].

PURPOSE: To develop an effective machine learning method for estimation of Ki-67 cell proliferation index. METHODS: Oral squamous cell carcinoma(OSCC) slices were selected for Ki-67 immunohistochemical staining. The digital pathology images were obtained through whole-slide imaging technology. Variable threshold method based on local statistics was applied to preprocess the images, aiming at reducing the noise in the images. Adaptive threshold method was used to remove the irrelevant light-colored background area in the image, retaining the nucleus part. A threshold method in space was applied to differentiate brown from blue content. Finally, the proliferation index was estimated and compared with manual and the color deconvolution method by paired sample t test and spearman correlation coefficients with SPSS 24.0 software package. RESULTS: A new nucleus detection and classification method was established, which can process pathologic images of different sizes, and effectively detect immunohistochemical brown positive cells and blue negative cells. There was no significant difference between this algorithm and manual counting（P＞0.05）, but the speed was faster. The calculation efficiency advantage was more obvious when processing a large image, and the detection result of Ki-67 proliferation index was better than the commonly used color deconvolution method（P＜0.05）. CONCLUSIONS: The automatic nucleus quantitative analysis method developed in this study can analyze Ki-67 staining of the nucleus in OSCC cells efficiently and calculate the proliferation index, which can be used for auxiliary diagnosis in pathology.

Surfactant-induced adsorption of Pb(II) on the cracked structure of microplastics.

Surfactant molecules can change the hydrophobic nature of microplastic surfaces, thereby affecting the adsorption of heavy metals in the environment onto the microplastics. It is essential to explore the role of crack structure of microplastics in the adsorption of heavy metals, especially in the presence of surfactants. In this study, polyethylene (PE) and polypropylene (PP) were evaluated for Pb(II) adsorption and desorption mechanism in the presence of two surfactants: cetyltrimethylammonium bromide (CTAB) and sodium dodecylbenzenesulfonate (SDBS). The experimental results were analyzed using kinetics and the isothermal model fitting and spectrogram (FTIR, XPS). This study showed that the application of surfactants could greatly enhance the Pb(II) adsorption capacities of PE and PP by promoting Pb(II) into the fissures. The Pb(II), S, and N contents did not significantly decrease at different depths in the presence of surfactants, and the Pb(II) content without surfactants decreased with an increasing depth. The adsorption behavior was consistent with the Bangham channel diffusion model and the DR model, suggesting that the adsorption process was related to the pore structure of the microplastics. Furthermore, the release of Pb(II) from desorption using high concentration of surfactant solution was less than that of low concentration, it was difficult to release heavy metals primarily because of the crack structure of the microplastics, especially when more surfactant molecules entered the pores. This work contributes to a better understanding of the adsorption mechanism of heavy metals on microplastics in the presence of surfactants, which will better control the ecological risks of microplastics.

Transport of different microplastics in porous media: Effect of the adhesion of surfactants on microplastics.

The adhesion of surfactant molecules on the microplastics surface is affected by the surface structure of the microplastics. Little is known about the mobility of different microplastics in the medium under surfactants. In order to reveal the migration of different microplastics under the action of surfactants, the study selected five kinds of microplastics (polyethylene (PE), polypropylene (PP), polystyrene (PS), polytetrafluoroethylene (PTFE), polymethyl methacrylate (PMMA)) and two kinds of surfactants (cetyltrimethylammonium bromide, CTAB and sodium dodecyl benzene sulfonate, SDBS) as the research objects. The column experiment was used to explore the transport behavior of microplastics under different concentrations of surfactants and the convection dispersion model was used to simulate. The dynamic contact angle of the surfactant solution on the microplastics was measured and the adhesion work was calculated by the Young-Dupre equation to reveal the underlying mechanism of microplastics retention in the presence of surfactants. The results showed that the transport ability of microplastics followed the order of PTFE

Comparison of Endoscopy Alone with Surgery Converted from Endoscopy for the Removal of Esophageal Foreign Bodies in Adults: A Retrospective Study from a Single Center.

BACKGROUND The impaction of an esophageal foreign body is an urgent situation requiring emergency intervention. This retrospective study from a single center in China aimed to compare endoscopy alone with surgery converted from endoscopy for the removal of esophageal foreign bodies in adults. MATERIAL AND METHODS A total of 252 patients with esophageal foreign bodies were divided into 3 groups based on the treatment received: endoscopy, surgery converted from endoscopy, or surgery only. Patients' clinical and demographic data were retrospectively reviewed and analyzed. RESULTS The diameter of the foreign bodies in patients treated by surgery converted from endoscopy was larger than that of those treated by simple endoscopy (5.2 cm vs 2.7 cm, P=0.0003). The cervical or upper thoracic esophagus was the most common site of foreign body impaction treated by surgery converted from endoscopy, while the foreign bodies removed by simple endoscopy were frequently lodged at the middle thoracic esophagus (P=0.021). Bone-related foreign bodies and dentures were most likely impacted in patients treated with surgery converted from endoscopy. The factors influencing the choice of treatment included foreign body maximal diameter and location. CONCLUSIONS Larger foreign bodies that were found in the cervical or upper thoracic esophagus were associated with failed endoscopic removal and required surgical removal. Irregularly shaped or sharp foreign bodies, including dentures and fishbones, required surgical removal. These findings may guide future decisions of first-line approaches for the removal of esophageal foreign bodies.

Surgical treatment of giant mediastinal tumors.

BACKGROUND: This study aims to evaluate the surgical treatment outcomes of giant mediastinal tumors. METHODS: Between July 2013 and July 2018, medical data of a total of 31 patients (26 males, 5 females; mean age 27.7±8.2 years; range, 18 to 56 years) who underwent radical surgery for a giant mediastinal tumor in our center and 47 cases (26 males, 21 females; mean age 45.4±16.7 years; range, 19 to 62 years) of giant mediastinal tumors retrieved from the National Center for Biotechnology Information database were retrospectively reviewed. Two-year overall survival and disease-free survival rates of the patients were evaluated. RESULTS: All patients underwent radical surgery (R0 resection). Symptoms caused by giant mediastinal tumors were relieved after radical surgery during follow-up. The two-year overall survival and disease-free survival rates were 100% and 86.7%, respectively, indicating a good prognosis. The surgical procedures for malignancies were more difficult than those for benign pathologies. CONCLUSION: Radical surgery is the mainstay for treatment of giant mediastinal tumors to relieve symptoms in a short period of time and to achieve a good prognosis for up to two years, regardless of adjuvant therapy. The surgical route should be cautiously planned before radical surgery to reduce complications.

Video-assisted thoracoscopic surgery is safe and reliable for large and invasive primary mediastinal tumors.

INTRODUCTION: Video-assisted thoracoscopic surgery (VATS) was not considered for the treatment of primary mediastinal tumors of large sizes or with local invasion. AIM: To investigate the clinical outcomes of VATS for large and invasive mediastinal tumors. MATERIAL AND METHODS: One hundred and thirteen patients with primary mediastinal tumors were treated by VATS. Twenty-nine patients had bulky tumors (diameter > 6 cm) and 5 patients had invasive tumors. Clinical data were documented and compared. RESULTS: No patients suffered from any complications after VATS. No relapse or metastasis occurred in the patients with bulky tumors, while 1 patient with invasive thymoma suffered a relapse after VATS. The 2-year disease-free survival and overall survival in patients with bulky tumors were 100% and 100%, while those in patients with invasive tumors were 75% and 100%. There were no differences in hospital stay after VATS between the patients with bulky tumors and smaller tumors, nor between the patients with invasive tumors and non-invasive tumors. Patients with bulky tumors lost more blood than those with smaller tumors, while more blood loss occurred in patients with invasive tumors than non-invasive tumors. Longer operative time was needed for patients with bulky tumors and invasive tumors. Mediastinal tumors with large size or invasion should not be contraindicated for VATS. The prognosis of such patients treated with VATS was comparable to that of traditional open surgery. CONCLUSIONS: VATS is a safe and effective procedure for large and invasive mediastinal tumors.

Long noncoding RNA TMPO-AS1 promotes progression of non-small cell lung cancer through regulating its natural antisense transcript TMPO.

Long noncoding RNAs (lncRNAs), a large group of RNAs with limited or no protein-coding capacity, have been demonstrated to play critical roles in human malignancy. The aim of this study is to examine the expression and function TMPO antisense transcript 1 (TMPO-AS1) in non-small cell lung cancer (NSCLC). Here, we found that the expression of both TMPO-AS1 and TMPO mRNA levels were overexpressed in NSCLC cells lines and tissues. A significant positive correlation was observed between TMPO-AS1 and TMPO mRNA levels. The upregulation of TMPO-AS1, TMPO mRNA and protein levels were associated with tumor progression of NSCLC. More importantly, through antisense pairing with TMPO mRNA, TMPO-AS1 regulates TMPO mRNA stability. Knockdown of TMPO-AS1 decreased the mRNA and protein levels of TMPO in NSCLC cells. An overlapping (OL) region was found between TMPO-AS1 and TMPO exon 1 and overexpression of TMPO-AS1-OL elevated the mRNA and protein levels of TMPO. Functionally, the downregulation of TMPO-AS1 significantly inhibited cells proliferation, colony formation, migration and invasiveness in vitro, and tumor growth in vivo. In contrast, overexpression of TMPO could promote the aggressive behaviors of NSCLC cells in vitro. Our findings indicate that TMPO-AS1 contributes to lung carcinogenesis, which may be partially through upregulation TMPO.

Low expression of occludin: a predictor of poor prognosis in esophageal squamous cell carcinoma.

Occludin is transmembrane protein and a key constituent of tight junction, and might participate in barrier function and fence function of epithelia and endothelia. It has been shown to be aberrantly expressed in malignant tumors and plays a role in carcinogenesis and tumor progression. The prognostic significance of Occludin expression has been implicated in various human cancers. However, the prognostic significance of Occludin expression in esophageal squamous cell carcinoma (ESCC) has not been established. In this study, we screened the tight junction genes aberrantly expressed based on two published gene microarray datasets (GSE20347 and GSE23400), and examined 95 esophageal cancer cases to assess immunohistochemical expression patterns of Occludin based on tissue array. Down-regulation of Occludin expression was shown in ESCC as compared with adjacent non-neoplastic specimens (P = 0.003). Decreased expression of Occludin was correlated with high histological grade (P = 0.017). Decreased expression of Occludin was also correlated with short overall survival (P = 0.014). The results indicated Loss of Occludin expression was associated with poor prognosis in ESCC, and Occludin expression was potentially a good predictor of prognosis in ESCC.

[Inhibitory effect of cyclopamine on metastatic ability of EC109 cells and its mechanism].

OBJECTIVE: To investigate the effect of cyclopamine on metastatic ability of human esophageal cancer EC109 cells and explore the possible mechanism. METHODS: Transwell chamber assay and angiogenesis assay were used to examine the metastatic ability, invasiveness and angiogenesis of EC109 cells treated with cyclopamine for 48 h. The expression of Gli-1 mRNA was detected using RT-PCR, and Western blotting was used to examine the protein expressions of Gli-1, matrix metalloproteinase-9 (MMP-9) and vascular endothelial growth factor (VEGF). RESULTS: Inhibition of the hedgehog signaling pathway by cyclopamine suppressed the migration, invasion, and angiogenesis of EC109 cells. Cyclopamine treatment significantly lowered the expression of Gli-1 mRNA (P<0.05) and the protein expressions of Gli-1, MMP-9 and VEGF (P<0.05). CONCLUSION: Cyclopamine can significantly inhibit the metastatic capacity of EC109 cells possibly by down-regulating MMP-9 and VEGF expression as a result of Gli-1 inhibition.

[Gene expression signature for lymphatic metastasis of human lung adenocarcinoma].

BACKGROUND AND OBJECTIVE: Distant metastasis is a major cause of mortality for patients with lung adenocarcinoma. So far, the mechanism of tumor metastasis is unknown. This study was to screen the gene expression signature in relation to lymphatic metastasis of human lung adenocarcinoma. METHODS: Primary lung adenocarcinoma tissues and regional lymph nodes were obtained from 22 patients underwent radical resection. The samples were classified into three groups: 11 cases of primary lung adenocarcinoma without lymphatic metastasis (TxN-), 11 cases of primary lung adenocarcinoma with lymphatic metastasis (TxN+), and 11 cases of the corresponding tumor cells from metastatic lymph nodes(N+). Total RNA was extracted from laser microdissected tumor samples. Adequate RNA starting materials from the primary tumors or metastatic nodes were labeled and then hybridized into the same microarray containing 6000 known human genes or expressed sequence tags (ESTs). After scanning, data analyses were performed using GeneSpring 6.2. RESULTS: Among 17 differentially expressed genes between the TxN+ and TxN-groups, 12 genes were significantly elevated and five genes were significantly downregulated in the TxN+ group compared with the TxN-group. There were 53 differentially regulated genes between the N+ and TxN+ groups, among which 25 genes were overexpressed and 28 genes were suppressed in the N+ group. CONCLUSION: The combination of early oncogenic alterations and later acquisition of a set of genetic alterations may determine the metastatic potential of lung adenocarcinoma.

Genetic fingerprint concerned with lymphatic metastasis of human lung squamous cancer.

BACKGROUND: With the most recent introduction of microarray technology to biology, it becomes possible to perform comprehensive analysis of gene expression in cancer cell. In this study the laser microdissection technique and cDNA microarray analysis were combined to obtain accurate molecular profiles of lymphatic metastasis in patients with lung squamous cell carcinoma. METHODS: Primary lung squamous cancer tissues and regional lymph nodes were obtained from 10 patients who underwent complete resection of lung cancer. According to the source of lung cancer cells, the samples were classified into three groups: the primary tumor with lymphatic metastasis (TxN+, n=5), the primary tumor without lymphatic metastasis (TxN-, n=5) and matched tumor cells from metastatic lymph nodes (N+, n=5). Total RNA was extracted from laser microdissected tumor samples. Adequate RNA starting material of mRNA from primary tumor or metastatic nodes were labeled and then hybridized into the same microarray containing 6 000 known, named human genes/ESTs. After scanning, data analysis was performed using GeneSpring6.2. RESULTS: A total of 37 genes were found to be able to separate TxN+ from TxN-. TxN+ have higher levels of genes concerned with structural protein, signal transducer, chaperone and enzyme. TxN- have higher levels of genes coding for cell cycle regulator, transporter, signal transducer and apoptosis regulator. Interestingly, there were no differentially expressed genes between N+ and TxN+. CONCLUSIONS: The acquisition of the metastatic phenotype might occur early in the development of lung squamous cancer. We raise the hypothesis that the gene-expression signature described herein is valuable to elucidate the molecular mechanisms regarding lymphatic metastasis and to look for novel therapeutic targets.