Long-term Proton Pump Inhibitor Administration Caused Physiological and Microbiota Changes in Rats.

Proton pump inhibitors (PPIs) are used for the long-term treatment of gastroesophageal disorders and the non-prescription medicines for acid reflux. However, there is growing concerns about PPI misuse, overuse and abuse. This study aimed to develop an animal model to examine the effects of long-term use of PPI in vivo. Twenty one Wistar rats were given omeprazole orally or intravenously for 30 days, and caerulein as a positive control. After euthanization, the serum and stool were collected to perform MS-based quantitative analysis of metabolites. We carried out 16S-based profiling of fecal microbiota, assessed the expression of bile acid metabolism regulators and examined the immunopathological characteristics of bile ducts. After long-term PPI exposure, the fecal microbial profile was altered and showed similarity to those observed in high-fat diet studies. The concentrations of several metabolites were also changed in various specimens. Surprisingly, morphological changes were observed in the bile duct, including ductal epithelial proliferation, micropapillary growth of biliary epithelium, focal bile duct stricture formation and bile duct obstruction. These are characteristics of precancerous lesions of bile duct. FXR and RXRα expressions were significantly reduced, which were similar to that observed in cholangiocarcinoma in TCGA and Oncomine databases. We established a novel animal model to examine the effects of long-term use of omeprazole. The gut microbes and metabolic change are consequences of long-term PPI exposure. And the results showed the environment in vivo tends to a high-fat diet. More importantly, we observed biliary epithelial hyperplasia, which is an indicator of a high-fat diet.

Monitoring and Determining Mitochondrial Network Parameters in Live Lung Cancer Cells.

Mitochondria are dynamic organelles that constantly fuse and divide, forming dynamic tubular networks. Abnormalities in mitochondrial dynamics and morphology are linked to diverse pathological states, including cancer. Thus, alterations in mitochondrial parameters could indicate early events of disease manifestation or progression. However, finding reliable and quantitative tools for monitoring mitochondria and determining the network parameters, particularly in live cells, has proven challenging. Here, we present a 2D confocal imaging-based approach that combines automatic mitochondrial morphology and dynamics analysis with fractal analysis in live small cell lung cancer (SCLC) cells. We chose SCLC cells as a test case since they typically have very little cytoplasm, but an abundance of smaller mitochondria compared to many of the commonly used cell types. The 2D confocal images provide a robust approach to quantitatively measure mitochondrial dynamics and morphology in live cells. Furthermore, we performed 3D reconstruction of electron microscopic images and show that the 3D reconstruction of the electron microscopic images complements this approach to yield better resolution. The data also suggest that the parameters of mitochondrial dynamics and fractal dimensions are sensitive indicators of cellular response to subtle perturbations, and hence, may serve as potential markers of drug response in lung cancer.

Expression of glypican 3 enriches hepatocellular carcinoma development-related genes and associates with carcinogenesis in cirrhotic livers.

Glypican-3 (GPC3) protein expression was determined by immunohistochemical analysis from 29 normal livers, 80 cirrhotic livers sample taken near hepatocellular carcinoma (HCC), and 87 cirrhotic livers without HCC. The levels for miR-657 and HCC-related gene mRNAs were determined by quantitative real-time polymerase chain reaction (qRT-PCR). Also, a published microarray dataset was used for gene set enrichment analysis (GSEA) to investigate the relationship between GPC3- and HCC-related gene signatures. Kaplan-Meier analysis was used to evaluate the relationship between GPC3 and HCC recurrence. GPC3 protein expression was not detected in any of the 29 (0%) normal livers, but was detected in 32 of 87 (37%) cirrhotic livers without HCC, and 51 of 80 (64%) cirrhotic liver samples taken near HCC sites (P < 0.001). The GPC3-positive rate in cirrhotic livers of viral origin was 68% (27/40), which was significantly higher than for non-viral cirrhotic livers (11%, 5/47) (P < 0.001). Also, GPC3 expression positively correlated with mRNA expression of HCC-related genes in the qRT-PCR and GSEA evaluations. Furthermore, HCC recurrence in cirrhotic liver samples taken near HCC sites was significantly higher in the GPC3-positive group than the GPC3-negative group (Log-rank P = 0.02, HR = 3.26; 95% CI = 1.20-10.29). This study demonstrated that highly expression of GPC3 could enrich HCC-related genes' mRNA expression and positive associate with dysplasia in cirrhotic livers. Therefore, GPC3 may serve as a precancerous biomarker in cirrhotic livers.

Merkel cell carcinoma of lymph node with unknown primary has a significantly lower association with Merkel cell polyomavirus than its cutaneous counterpart.

Rare cases of Merkel cell carcinoma have been encountered in lymph nodes with unknown extranodal primary, which exhibit similar morphologic and immunophenotypic features to those in primary cutaneous Merkel cell carcinomas. However, it is uncertain whether the nodal Merkel cell carcinoma is a primary tumor of the lymph node or represents a metastasis from an occult or regressed extranodal lesion. To establish an accurate diagnosis of the nodal Merkel cell carcinoma can be challenging because of significant morphologic mimics, including lymphoblastic lymphoma and metastatic small cell carcinoma. Moreover, there is no consensus for a diagnostic term, and many different terms have been used, which can be confusing and may not fully reflect the nature of nodal Merkel cell carcinoma. In this study, we investigated the detailed clinicopathologic features of 22 nodal Merkel cell carcinomas, with comparison to 763 primary cutaneous cases retrieved from the literature. Overall, the nodal and cutaneous Merkel cell carcinomas shared similar clinical presentations, morphologic spectrum, and immunophenotype; both were mostly seen in elderly male with a typical neuroendocrine morphology. Most of cases expressed CK20, synaptophysin, and chromogranin A; and PAX5 and TdT were also positive in majority of cases. However, nodal Merkel cell carcinomas had a significantly lower association with Merkel cell polyomavirus than cutaneous cases (31% vs 76%, P=0.001). Therefore, these two entities may arise from overlapping but not identical biological pathways. We also recommend the use of the diagnostic term 'Merkel cell carcinoma of lymph node' to replace many other names used.

Napsin A expression in primary mucin-producing adenocarcinomas of the lung: an immunohistochemical study.

Immunohistochemical expression of napsin A in primary pulmonary mucinous tumors is not well established. Napsin A immunoreactivity was evaluated in 43 mucin-producing adenocarcinomas of the lung consisting of 18 tumors formerly classified as mucinous bronchioloalveolar carcinoma, 15 colloid adenocarcinomas, 5 solid predominant adenocarcinomas with mucin production, and 5 adenocarcinomas with signet ring cell features, as well as in 25 extrapulmonary mucinous adenocarcinomas of different anatomic sites. Immunohistochemical expression of thyroid transcription factor 1 (TTF-1) was also compared. Thirty-three percent of mucinous lung tumors exhibited positive immunoreactivity for napsin A, whereas 42% expressed TTF-1. All 25 extrapulmonary mucinous adenocarcinomas lacked expression of napsin A and TTF-1. Mucin-producing neoplasms of the lung infrequently express napsin A, suggesting that immunohistochemical assessment of napsin A may have limited diagnostic usefulness for distinguishing primary and metastatic mucinous adenocarcinomas involving the lung.

An overview of the rare parotid gland cancer.

Cancer of the parotid gland is relatively rare, but carries poor prognosis owing to its prevailing distant metastases. In addition to the disease's basic epidemiology and pathology, we review some current discoveries of its tumorigenesis molecular mechanism. Based on published salivary gland cancer clinical trial data, non-surgical antitumor efficacies amongst a range of chemotherapy, radiation, and concurrent therapy regimens are compared. We also present the current development status of novel radiation therapy and targeted therapeutics, focusing on intensity-modulated radiation therapy (IMRT), and epidermal growth factor receptor (EGFR) and vascular endothelial growth factor (VEGF) blockages, which are showing promise for improving parotid gland cancer management.

Determining the site of origin of mucinous adenocarcinoma: an immunohistochemical study of 175 cases.

Mucinous adenocarcinomas (MAs) of various origins may have a similar histologic appearance and frequently metastasize to distant sites, which often causes diagnostic problems in surgical pathology practice. The immunohistochemical profiles of MAs of various origins have not been well studied. We investigated the expression of 10 immunohistochemical markers (CK7, CK20, CDX-2, ß-catenin, MUC-1, MUC-2, MUC-6, ER, WT-1, and PAX-8) in 175 cases of MA, including 69 cases from the lower gastrointestinal (GI) tract, 41 from the upper GI tract, 27 from gynecologic organs, 4 from the urinary bladder, 18 from the breast, and 16 from the lung. We found that lower GI MAs (colon, rectum, and anus) frequently expressed CDX-2 (42 of 42, 100%; 33 of 42 with homogenous positivity, 79%), MUC-2 (42 of 42; 100%), CK20 (41 of 42; 98%), and ß-catenin (nuclear) (27 of 42; 64%) and rarely expressed MUC-6 (2 of 42; 5%) and CK7 (8 of 42; 19%). Most of the CK7-positive cases were from the rectum and anus (7 of 8; 88%). The expression of these markers in appendiceal MAs was similar to that of low GI tract MAs, except for a lower percentage of homogenous CDX-2 (3 of 27; 11%) and nuclear ß-catenin (3 of 27; 11%) expression. Unlike their lower GI tract counterparts, the upper GI tract MAs (ampulla, pancreas/biliary tree, and stomach/esophagus) frequently expressed CK7 (38 of 41; 93%) and MUC-6 (31 of 41; 76%) and were rarely homogenously positive for CDX-2 (4 of 41; 10%) and nuclear positive for ß-catenin (8 of 41; 19%). Breast MAs were frequently positive for CK7 (18 of 18; 100%), MUC-1 (18 of 18; 100%), MUC-2 (18 of 18; 100%), ER (16 of 18; 89%), MUC-6 (9 of 18; 50%), and WT-1 (9 of 18; 50%). Lung MAs were frequently positive for CK7 (16 of 16; 100%) and MUC-1 (15 of 16; 94%). Gynecologic MAs were positive for CK7 (25 of 27; 93%) and PAX-8 (13 of 27; 48%). We conclude that homogenous CDX-2 and nuclear ß-catenin expressions are commonly seen in lower GI tract MAs. In contrast, appendiceal MAs are usually heterogenously positive for CDX-2 and show cytoplasmic positivity for ß-catenin. Unlike lower GI tract MAs, upper GI tract MAs are frequently positive for CK7 and MUC-6. As is the case in appendiceal MAs, the upper GI tract MAs may also be heterogenously positive for CDX-2. Breast MAs are positive for ER and WT-1, whereas gynecologic MAs are positive for PAX-8 and negative for WT-1.

Alloimmune response results in expansion of autoreactive donor CD4+ T cells in transplants that can mediate chronic graft-versus-host disease.

Chronic graft-versus-host disease (cGVHD) is considered an autoimmune-like disease mediated by donor CD4(+) T cells, but the origin of the autoreactive T cells is still controversial. In this article, we report that the transplantation of DBA/2 donor spleen cells into thymectomized MHC-matched allogeneic BALB/c recipients induced autoimmune-like cGVHD, although not in control syngeneic DBA/2 recipients. The donor-type CD4(+) T cells from the former but not the latter recipients induced autoimmune-like manifestations in secondary allogeneic BALB/c as well as syngeneic DBA/2 recipients. Transfer of donor-type CD4(+) T cells from secondary DBA/2 recipients with disease into syngeneic donor-type or allogeneic host-type tertiary recipients propagated autoimmune-like manifestations in both. Furthermore, TCR spectratyping revealed that the clonal expansion of the autoreactive CD4(+) T cells in cGVHD recipients was initiated by an alloimmune response. Finally, hybridoma CD4(+) T clones derived from DBA/2 recipients with disease proliferated similarly in response to stimulation by syngeneic donor-type or allogeneic host-type dendritic cells. These results demonstrate that the autoimmune-like manifestations in cGVHD can be mediated by a population of donor CD4(+) T cells in transplants that simultaneously recognize Ags presented by both donor and host APCs.

Mixed cortical adenoma and composite pheochromocytoma-ganglioneuroma: an unusual corticomedullary tumor of the adrenal gland.

Adrenal neoplasms composed of more than one cell type and demonstrating a mixed histologic appearance are exceedingly rare. We report the clinical and pathologic features of a morphologically distinctive tumor of the adrenal gland composed of cortical, chromaffin, and neural cells. Histologically, the tumor consisted of intermixed areas of proliferating cortical cells resembling adrenal cortical adenoma, neoplastic chromaffin cells consistent with pheochromocytoma, and a ganglioneuromatous stroma. The presence of the cortical, medullary, and neural components within the tumor was confirmed by immunohistochemical studies. The present case serves to broaden the morphologic spectrum of mixed tumors that may be encountered in the adrenal gland.

Myopericytoma of the kidney.

Myopericytoma is a rare, histologically distinctive tumor that shows evidence of differentiation toward perivascular myoid cells. Myopericytoma is largely considered a neoplasm of skin and soft tissues, with examples of this lesion involving visceral sites being extremely limited. We present the clinical and pathologic details of an unusual case of myopericytoma occurring in the kidney. Histologically, the tumor was richly vascularized and composed of a perivascular proliferation of oval to spindle-shaped cells with bland cytologic features. The neoplastic cells were arranged in a concentric fashion around vascular lumina and also surrounded dilated, branching vessels, with a glomangiopericytomatous appearance. Mitotic figures were inconspicuous, and necrosis was absent. Perivascular myoid differentiation was supported by positive immunoreactivity for muscle-specific and smooth muscle actins, and absence of reactivity for desmin. The present case serves to expand the anatomical distribution of myopericytoma and also broadens the spectrum of primary mesenchymal neoplasms that may be encountered in the kidney.

TCL1 protein expression in testicular germ cell tumors.

We immunohistochemically studied TCL1 protein expression in different histologic types of 63 testicular germ cell tumors: 23 seminomas, 14 embryonal carcinomas, 4 teratomas, 2 yolk sac tumors, and 20 mixed germ cell tumors. The 20 mixed germ cell tumors contained components of seminoma (n = 10), embryonal carcinoma (n = 18), teratoma (n = 16), yolk sac tumor (n = 7), and choriocarcinoma (n = 3). We also examined 40 cases of intratubular germ cell neoplasia, unclassified type (IGCNU). Positive immunoreactivity for TCL1 was observed in 91% of the seminoma samples (30/33) and all IGCNU cases. In contrast, no TCL1 expression was detected among the nonseminomatous germ cell tumors. In the context of testicular germ cell neoplasia, the presence of TCL1 protein appears restricted to IGCNU and seminoma, suggesting association with an undifferentiated state and loss of protein expression with tumor differentiation. Immunohistochemical evaluation of TCL1 expression may have usefulness in substantiating a diagnosis of IGCNU or seminoma and in the separation of seminoma from nonseminomatous germ cell tumors.

A newly identified variation at the entry of the recurrent laryngeal nerve into the larynx.

OBJECTIVES: We aimed to highlight a new anatomical variation of the recurrent laryngeal nerve (RLN), and to emphasize its implications for thyroid surgery. METHODS: A prospective study was carried out in a group of 3,078 consecutive thyroidectomies from 1998 to 2008. Total, near-total, subtotal, and partial thyroidectomy were performed for various thyroid diseases. The RLN was routinely identified and exposed in its entire course until the entry into the larynx. The postoperative complications of patients with different variations were compared. RESULTS: 4,241 RLNs were successfully identified in all patients unilaterally or bilaterally. In addition to extralaryngeal branching and nonrecurrent laryngeal nerves, an unreported variation was identified in 44 RLNs (1.04%) at their entries into the larynx. The variation happened at the trunk or the branches of the RLN entering the larynx far from the posterior of cricothyroid joint, and the entry was higher than the superior cornu of the thyroid cartilage and the arch of the cricoid. The median distance from the entry to the posterior of cricothyroid joint was more than 5 mm. As the trunk or the branches had to travel along the lateral edge of the upper 1/3 of the thyroid before entering the larynx, the incidence of RLN palsy was higher than that in extralaryngeal branching variations (p < .05). CONCLUSION: This newly discovered variation of the RLN is more vulnerable to injury and should be brought to the attention of surgeons.

Amyand's hernia in elderly patients: diagnostic, anesthetic, and perioperative considerations.

The presence of a vermiform appendix in an inguinal hernia sac is termed as Amyand's hernia. Although rare, mistakes in diagnosis and treatment can cause catastrophic results. Charts of patients with inguinal hernia were reviewed, and four cases of Amyand's hernia were confirmed. The clinical presentation, anesthetic, and perioperative management of Amyand's hernia were further analyzed. The mean age of patients was over 70 years, and all were males. None of the patients were diagnosed preoperatively. All the patients had little abdominal complaint only with a right inguinal mass and dragging sensation for several hours. Due to the short time after incarceration and significant cardiovascular and pulmonary comorbidities, manual reduction was attempted first in three patients. With complete preoperative evaluation and careful perioperative support, all patients underwent appendectomy and Bassini's hernia repair through a groin incision. Based on age-related organ failure and associated chronic medical illnesses of geriatric patients, the difficulties in the diagnosis and treatment are also summarized and analyzed.

The plication and splinting procedure for idiopathic sclerosing encapsulating peritonitis.

Idiopathic sclerosing encapsulating peritonitis (ISEP) is a rare cause of small intestinal obstruction. Histologically, ISEP is characterized by a thick fibrotic membrane encasing the small bowel without any apparent pathophysiological factors. While ISEP is predominantly present congenitally in female adolescents from subtropical region, it has been identified throughout the world. Evidence-based effective diagnostics and treatments are pitifully thin. We experienced six cases, four males and two females, which exhibited symptoms in their later thirties and forties. Five patients presented with acute and subacute intestinal obstruction, and one patient with cryptorchidism and seminoma was referred. Due to the limitation in distention and motility of bowel loops of ISEP, imaging exams may not be very reliable for accurate diagnosis and estimation of obstruction progress. They were successfully treated with the mesenteric plication and intraluminal splinting procedures. Two cases had an uneventful postoperative period, and the returns of normal bowel function were delayed in the other four patients. Two recurrences of small bowel obstructions were noted over a mean follow-up period of 33 months with mild symptoms. This article reviews the patterns of clinical presentations, diagnostic clues, and theories of potential risk factors of ISEP as well as its controversial surgical managements.

Intestinal type of mucinous borderline tumor arising from mixed epithelial and stromal tumor of kidney.

We report a mucinous borderline tumor arising from a mixed epithelial and stromal tumor of left kidney (MESTK). The patient was an 82-year-old woman who presented with gross hematuria and recurrent urinary tract infection for years. The patient had a cytoscopy with a retrograde pyelogram, which indicated a dilated left kidney with a central mass lesion. Subsequently, she underwent a radical left nephrectomy. Cross-sections of left kidney showed a 4.5 x 3.5 x 1.5 cm ill-defined cystic lesion with mucinous and solid areas. Histologic sections of the lesion showed numerous variable-sized dilated cysts with fibrous, fatty, vascular, and smooth muscle stroma. The cysts were lined by a various types of epithelium, including single layer of flat, cuboidal and mucinous epithelium, urothelium, intestinal epithelium, and endocervical epithelium. In areas, the mucinous epithelium showed complex proliferation with stratification, papillae formation, and nuclear atypia, resembling that of an ovarian mucinous borderline tumor, a colorectal tubular adenoma, or a low-grade appendiceal mucinous carcinoma. Immunohistochemically, the mucinous borderline tumor showed a colorectal phenotype, being strongly positive for CK20, CDX-2, and MUC2. There was no invasive mucinous tumor identified. We believe that this case represents the first reported example of mucinous borderline tumor arising from a MESTK.

CC chemokine receptor 9 enhances proliferation in pancreatic intraepithelial neoplasia and pancreatic cancer cells.

INTRODUCTION: Chemokine receptors may regulate the progression and metastasis of invasive malignancies. There are little data, however, regarding their role in premalignant lesions. Our objective was to determine the role of CC chemokine receptor 9 (CCR9) in pancreatic intraepithelial neoplasia (PanIN). METHODS: Human and murine formalin-fixed paraffin-embedded (FFPE) PanIN specimens were assessed for CCR9 expression. The established murine PanIN, invasive pancreatic cancer (5143PDA) and liver metastasis (5143LM) cell lines, and human pancreatic cancer cell line (PANC-1) were obtained to verify CCR9 expression and function. RESULTS: Immunohistochemistry of FFPE specimens demonstrated CCR9 expression in both murine and human PanIN lesions. CCR9 expression in murine and human cell lines was verified by Western blot assay, immunofluorescence, and flow cytometry. CCR9 function was demonstrated by in vitro exposure to CCL25, the selective CCR9 ligand, which resulted in significantly increased cell proliferation in PanIN and pancreatic cancer cell lines. CONCLUSIONS: This is the first report of chemokine receptor CCR9 expression in murine and human PanIN tissues. Our results demonstrate enhanced PanIN and pancreatic cancer cell proliferation with activation of CCR9 by its selective ligand CCL25. CCR9 may prove to be a novel therapeutic target for PanIN and its progression to invasive cancer.

Overlapping features between dedifferentiated liposarcoma and undifferentiated high-grade pleomorphic sarcoma.

Dedifferentiated liposarcoma (DDL), occurring in up to 10% of well differentiated liposarcoma cases, has similar histologic features to that of undifferentiated high-grade pleomorphic sarcoma (UHGPS); the former develops in a background of atypical lipomatous tumors/well differentiated liposarcoma, whereas the latter shows no specific line of differentiation. The retroperitoneum and thigh represent the most common anatomic locations for both the sarcomas. Despite their morphologic similarity, the issue of whether these 2 sarcomas share overlapping immunohistochemical and molecular features has not been well studied. We examined the expression of the lipogenic tumor-related markers peroxisome proliferator-activated receptor gamma (PPAR-gamma), CDK4, and MDM2 in 15 cases of DDL and 45 cases of retroperitoneal/thigh UHGPS. Patients with DDL ranged from 31 to 82 years (mean 63 y) with a male:female ratio of 5:3. Patients with UHGPS ranged from 14 to 80 years (mean 52 y) with a male:female ratio of 3:2. All 15 DDLs expressed CDK4 and MDM2 (100%), and 8 of 15 cases expressed PPAR-gamma (53%). Twenty-three of 45 (51%) UHGPS expressed at least 1 of these 3 markers. We also studied MDM2 and CDK4 gene amplification by fluorescence in situ hybridization in 28 immunohistochemically positive cases, including 5 DDLs and 23 UHGPSs. All 5 cases of DDL showed MDM2 and/or CDK4 amplification (100%), whereas 6 of 45 UHGPSs showed MDM2 and/or CDK4 amplification (13%). Our results demonstrate that (1) the lipogenic tumor markers CDK4 and MDM2 can be used as surrogate immunohistochemical markers for the diagnosis of malignant lipomatous tumors with high sensitivity; (2) approximately 26% of retroperitoneal/thigh UHGPS cases that were positive for PPAR-gamma, CDK4, or MDM2 by immunohistochemistry showed characteristic CDK4 and MDM2 gene amplification, suggesting that a subset of UHGPS cases represent DDL despite lacking histologic evidence of lipoblasts.

The feasibility of total or near-total bilateral thyroidectomy for the treatment of bilateral multinodular goiter.

OBJECTIVE: To evaluate the feasibility and safety of total and near-total bilateral thyroidectomy for the treatment of bilateral multinodular goiter. METHODS: 346 patients with a diagnosis of bilateral multinodular goiter were randomly divided into two groups. 165 patients underwent total thyroidectomy or near-total thyroidectomy (group A), while 181 patients were exposed to a partial or subtotal thyroid gland removal treatment (group B). The incidences of postoperative complications and recurrence rate were monitored during the average follow-up period of 36 and 39 months, respectively. RESULTS: Six and two patients from groups A and B, respectively, were diagnosed with papillary carcinoma and excluded from the study. Transient recurrent laryngeal nerve paralysis occurred in three patients each from group A (1.89%, 3/159) and group B (1.68%, 3/179) postoperatively. Injury to superior laryngeal nerve was confirmed in three patients (two in group A and one in group B). Eleven (6.92%, 11/159) and nine (5.03%, 9/179) cases in groups A and B, respectively, suffered from transient hypocalcemia symptoms. There was no statistical difference in complications between two groups. Permanent hypoparathyroidism was not observed in either group. No recurrence was observed in group A, while 12 cases (6.70%, 12/179) were observed in group B. The recurrence rate was significantly different between the two groups (p <.05). CONCLUSION: It is safe and feasible to perform either total or near-total thyroidectomy in patients with bilateral multinodular goiter. These treatments provide decisive advantages over partial and subtotal thyroidectomies in terms of the recurrence and reoperation rate with comparable postoperative complications.