Estrogen-Induced Stromal FGF18 Promotes Proliferation and Invasion of Endometrial Carcinoma Cells Through ERK and Akt Signaling.

OBJECTIVE: The aim of this study was to evaluate whether estrogen promoted the proliferation and invasion of endometrial carcinoma (EC) cells through paracrine FGFs in endometrial stromal cells (ESCs). PATIENTS AND METHODS: We screened gene alterations in a primary ESC culture after 10 nM estrogen treatment using an Agilent mRNA microarray. We knocked down stromal FGF18 expression in a co-culture system and aimed to explore the contribution of E2-induced stromal FGF18 to the proliferation and invasion of EC cells. To determine the effective receptors and detailed downstream signaling of FGF18, we co-cultured estrogen-treated hESCs with FGFR1-, FGFR2-, FGFR3- or FGFR4-knockdown Ishikawa cells. Finally, we detected FGF18 expression in clinical samples, including several primary cultures of different ESCs and a series of tissue microarrays (TMAs) of 90 patients with EC. RESULTS: A few genes altered significantly in estrogen-treated primary ESCs, but only FGF18 was noticeably enhanced among the FGF family genes. Knockdown of FGF18 expression in hESCs inhibited the promoting effect of FGF18 on the proliferation and invasion of EC cells. FGF18 bound FGFR2 and FGFR3 in Ishikawa cells to activate downstream ERK and Akt pathways and to promote the viability of EC cells. The FGF18-FGFR2 and FGF18-FGFR3 pathways had close correlations with Survivin and CD44V6 expression but not with P53. Primary ESCs of endometrioid EC (EEC, type I EC) had higher FGF18 expression than ESCs of normal endometrium (NE), endometrial atypical hyperplasia (EAH) and type II EC. CONCLUSION: Estrogen induced FGF18 in ESCs to promote the proliferation and invasion of EC cells, and FGFR inhibitors should be considered as promising candidate targets for EC treatment.

Serum Calcium and Risk of Nonmedullary Thyroid Cancer in Patients with Primary Hyperparathyroidism.

BACKGROUND Clinical cases of nonmedullary thyroid carcinoma (NMTC) in combination with primary hyperparathyroidism (PHPT) have been reported occasionally. However, the clinical characteristics and risk factors of concomitant NMTC in PHPT patients remain unclear. This study aimed to assess the association between PHPT and NMTC, and evaluate the clinical characteristics and risk factors of NMTC in Chinese patients with PHPT. MATERIAL AND METHODS This was a retrospective cohort analysis. We reviewed the medical records of 155 patients who underwent surgery for PHPT in two large medical centers in China between 2009 and 2014. The clinical manifestations, biochemical abnormalities, and histological characteristics of PHPT patients were analyzed. RESULTS Of the 155 patients with PHPT, 58 patients (37.4%) had thyroid nodules and 12 patients (7.7%) were ill with concomitant NMTC. PHPT patients with NMTC demonstrated significantly lower preoperative serum calcium levels compared to PHPT patients with benign thyroid nodules (p<0.05). A significantly negative association between preoperative serum calcium levels and the presence of NMTC was found in PHPT patients (p<0.05). Furthermore, ROC analysis revealed that albumin-corrected serum calcium levels <2.67 mmol/L had good capacity to differentiate the PHPT patients with NMTC from those with benign thyroid nodules. CONCLUSIONS Compared with the reported much lower prevalence of thyroid carcinoma in the general population, our results suggest that PHPT might be a risk factor for the malignancy of thyroid nodules; a lower level of serum calcium may predict the existence of NMTC in PHPT patients with thyroid nodules.

An Intracranial Gliosis Mimicking Neoplasm: A Dilemma.

Intracranial gliosis has no typical clinical signals or imaging characteristics. Therefore, it can be easily misdiagnosed as neoplasm. Hereby, we report a unique case of gliosis that grew outward from the surface of the brain. MRI depicted its signal and enhancement pattern similar to the cerebral gray matter. The diagnosis was confirmed by pathology and immunohistochemistry. Although it was difficult to reach a diagnosis, correlating its origin, growing pattern and MR features and knowing that gliosis can present this way may help in differentiating it from other diseases.

Venous thromboembolism is a product in proliferation of cancer cells.

The pathogenesis of venous thromboembolism (VTE) in patients with cancer is related to the destruction of small veins and the intravenous formation of filamentous mesh-like structure by fibrinogen. The filamentous mesh-like filter can block hematogenous metastasis of cancer cells and also can stagnate blood cells, leading to venous thrombosis. Cancer cells have characteristics of malignancy and fast proliferation, and ischemic necrosis frequently occurs, and small veins were invaded and damaged. The formation of filamentous mesh-like structure has defense function and also may cause the occurrence of VTE. VTE is a product of the proliferation process of malignant cells.

Activation of circulated immune cells and inflammatory immune adherence are involved in the whole process of acute venous thrombosis.

OBJECTIVE: To investigate localization and distribution of integrin subunit ß1, ß2 and ß3 and morphological changes of ligand-recepter binding in thrombi of acute pulmonary embolism (PE) patients and explore activation of circulated immune cells, inflammatory immune adherence and coagulation response in acute venous thrombosis. METHODS: Thrombi were collected from patients with acute PE. Immunohistochemistry was done to detect the expression and distribution of integrin ß1, ß2 and ß3 in cells within thrombi, and ligands of integrin subunit ß1, ß2 and ß3 were also determined by immunohistochemistry within the thrombi. RESULTS: 1) Acute venous thrombi were red thrombi composed of skeletons and filamentous mesh containing large amounts of red blood cells and white blood cells; 2) Integrin subunit ß1, ß2 and ß3 were expressed on lymphocytes, neutrophils and platelets; 3) No expression of integrin ß1 ligands: Laminin, Fibronectin, Collagen I or Collagen-II on lymphocytes; integrin ß2 ligands including ICAM, factor X and iC3b are distributed on neutrophils, and ligand fibrinogen bound to neutrophils; integrin ß3 was expressed on platelets which form the skeleton of thrombi and bound to fibrinogen to construct mesh structure; 4) Factor Xa was expressed on the filamentous mesh; 5) Filamentous mesh was fully filled with red blood cell dominant blood cells. CONCLUSION: Acute venous thrombosis is an activation process of circulated lymphocytes, neutrophils and platelets mainly, and a whole process including integrin subunit ß2 and ß3 binding with their ligands. Activation of immune cells, inflammatory immune adherence and coagulation response are involved in the acute venous thrombosis.

[Effect of Wnt signaling suppression on gefitinib in non small cell lung cancer cell lines].

OBJECTIVE: To explore the effect of Wnt signaling suppression on proliferation of non small cell lung cancer to gefitinib, and its related mechanisms. METHODS: PC9 and PC9/AB2 cells of both gefitinib sensitive and resistant were treated with different concentrations of gefitinib, and the proliferation index was measured using CCK8 kit. The members of Wnt signaling pathway were detected by Western blot. Dual luciferase reportor gene assay (TOP Flash) was used to document the transcriptional level of ß-catenin. ß-catenin siRNA was transfected into PC9/AB2 cells to suppress the Wnt signaling transcription, followed by treatment with different concentrations of gefitinib. Western blot was then used to detect the expression of EGFR and its downstream signaling after inhibit the expression of ß-catenin. RESULTS: Treating with different concentrations of gefitinib, the resistance of PC9/AB2 cells to gefitinib was significantly increased (P < 0.05). The members of Wnt signaling expressed at higher level in PC9/AB2 cells than in PC9 cells (t = 24.590, P = 0.000). TOP Flash examination showed that the endogenous transcriptional activity of Wnt signaling was higher in PC9/AB2 cell than that in PC9 cell (t = 4.983, P = 0.008). Compared with the negative control group, apoptotic rate and sensitivity to gefitinib significantly increased in interfered group (P < 0.05). The expression of p-ERK1/2 significantly decreased after Wnt signaling suppression, although other proteins showed no significant alterations. CONCLUSION: Suppressing the activity of Wnt signaling can partly reverse the celluar resistance to gefitinib in non small cell lung cancer.

[Effects of 1, 25 (OH)2D3 on bleomycin-induced pulmonary fibrosis in mice].

OBJECTIVE: To observe the effects of 1, 25(OH)2D3 on bleomycin-induced pulmonary fibrosis in mice and to explore its mechanisms. METHODS: Ninety male C57BL/6 mice, 6 to 8 weeks old, were randomly divided into 3 groups according to the table of random numbers: a control group, a model group and a treatment group(n = 30 each). Bleomycin was injected to the mice in the latter 2 groups by single intratracheal injection to duplicate the pulmonary fibrosis model, while the control group was injected with saline. From the next day, the mice in the treatment group received 1, 25 (OH) 2D3 (0.5 µg·kg(-1)·d(-1)) diluted in olive oil by gavage daily, while the other groups were treated with equivalent olive oil. Ten mice in each group were killed randomly on day 14, 21 and 28 after surgery respectively. Pulmonary alveolitis and fibrosis were evaluated by using hematoxylin-eosin and Masson stain method. The content of hydroxyproline was measured by acid hydrolysis method. The mRNA levels of collagen1α1, α-SMA, Wnt3a, Wnt4, and Wnt7a in the lung tissues were measured by real-time RT-PCR, while the protein expression of α-SMA and ß-catenin were assessed by immunohistochemistry. RESULTS: Pulmonary alveolitis at day 14, 21 and fibrosis at day14, 21, 28 in the treatment group were remarkably reduced compared to the model group (all P < 0.05). Compared with the model group, the treatment group showed decreased content of hydroxyproline, decreased mRNA levels of collagen1α1, α-SMA, Wnt3a, Wnt4 and decreased protein expression of α-SMA and ß-catenin at the 3 time points (all P < 0.05). The content of hydroxyproline and the mRNA levels of collagen1α1, α-SMA, Wnt3a, Wnt4, Wnt7a in the treatment group at 28 d were 0.67 ± 0.14, 1.66 ± 0.34, 1.37 ± 0.41, 1.43 ± 0.27, 1.29 ± 0.19, 1.18 ± 0.20, respectively, all of which were significantly lower than those in the model group (1.10 ± 0.16, 3.50 ± 0.74, 2.68 ± 0.61, 2.60 ± 0.58, 2.23 ± 0.45, 1.93 ± 0.36, respectively). Protein expression of α-SMA and ß-catenin in the treatment group were 0.44 ± 0.13 and 0.25 ± 0.05, respectively, which were also significantly lower than those of the model group(0.98 ± 0.20, 0.58 ± 0.06, respectively). CONCLUSION: 1, 25 (OH) 2D3 was shown to reduce pulmonary fibrosis induced by bleomycin in mice, and its mechanisms might be associated with Wnt signaling suppression.

[The ultrastructural features of sputum deposition and its value in the diagnosis of pulmonary alveolar proteinosis].

OBJECTIVE: To investigate the ultrastructural features of sputum deposition (SD) and its value in the diagnosis of pulmonary alveolar proteinosis (PAP). METHODS: Seven patients with PAP diagnosed by lung biopsy and cytology were enrolled in this study. The patients consisted of 5 men and 2 women, whose median age was 48 years (range 36 to 73). SD and bronchoalveolar lavage fluid (BALF) sediment were made into ultrathin sections and observed under transmission electron microscope (TEM), respectively. Seven cases of control group composed of 4 men and 3 women whose median age was 49 years (range 39 to 68) including 3 cases of bacterial pneumonia, two cases of COPD and 2 cases of exudative pulmonary tuberculosis. Each SD was made into ultrathin section, and compared with the experimental group. RESULTS: In PAP group, Periodic acid-schiff (PAS) staining was performed on 7 sputum smears and none of them was tested positive for any components with diagnostic interest. Four cases from the 7 paraffin-embed sections of BALF sediment by microscopic examination suggested PAP. Under TEM, BALF sediment showed that many lamellar bodies existed in and outside alveolar epithelial cells, and 5 specimens were consistent with PAP diagnosis. Compared with BALF sediment, SD had apparent degeneration with more myelin phagosomes in the cytoplasm of macrophages, more lamellar bodies in alveolar epithelial cells, and lots of lamellar bodies in the shape of concentric circle in the extracellular spaces. Four from the 7 SD samples were consistent with the diagnosis of PAP. No significant difference was found between SD and BALF in the diagnosis of PAP by electronic examination (P > 0.05). In the 7 cases of control group no drifting osmiophilic lamellar bodies in extracellular space were detected. CONCLUSIONS: The osmiophilic lamellar bodies with diagnostic value were found in SD and BALF of patients with PAP. TEM of SD in combination with clinical manifestations and radiologic findings can make a definitive diagnosis of PAP, especially for those patients who have contraindications to lung biopsy and lung lavage.

Case of interstitial lung disease possibly induced by exposure to iron dust.

Interstitial lung diseases are primarily attributable to occupational or environmental exposures to dusts and irritants. We report on a case of interstitial lung disease, possibly secondary to iron exposure. Our male patient presented with cough and shortness of breath of more than 20 years' duration after his occupational exposure had ended. A chest radiograph showed patchy shadows throughout both lower fields, and computed tomography showed ground-glass-like opacification, with fibrosis in the lower lobes. A lung biopsy revealed foamy cells in the alveolar spaces, with bronchiolitis obliterans. Microelemental analysis showed an increased level of iron in the lung tissue. After treatment with N-acetyl cysteine effervescent tablets, the patient's symptoms gradually improved. This probable case of iron-induced interstitial lung disease suggests the importance of obtaining a patient's history of occupational and environmental exposures for the sake of an accurate diagnosis.

Malignant mesothelioma presenting as a giant chest, abdominal and pelvic wall mass.

Malignant mesothelioma (MM) is a relatively rare carcinoma of the mesothelial cells, and it is usually located in the pleural or peritoneal cavity. Here we report on a unique case of MM that developed in the chest, abdominal and pelvic walls in a 77-year-old female patient. CT and MRI revealed mesothelioma that manifested as a giant mass in the right flank and bilateral pelvic walls. The diagnosis was confirmed by the pathology and immunohistochemistry. Though rare, accurate investigation of the radiological features of a body wall MM may help make an exact diagnosis.

[A study on the efficacy of glucocorticoid therapy for idiopathic nonspecific interstitial pneumonia].

OBJECTIVE: To investigate the efficacy of glucocorticoid therapy in idiopathic nonspecific interstitial pneumonia (INSIP). METHODS: Twenty-nine cases of INSIP confirmed by clinical- radiological-pathological (CRP) diagnosis were collected and classified into 2 groups according to the degree of fibrosis: INSIP-1 (including 9 cases of cellular type) and INSIP-2 (20 cases of mixed and fibrotic type). Thirty cases of usual interstitial pneumonia (UIP) confirmed by CRP diagnosis served as the control. Clinical and pathological features, therapeutic effects of glucocorticoids and the follow-up results were retrospectively analyzed and the survival curves were evaluated by Kaplan-Meier method. RESULTS: The mean age at onset of INSIP-1 group [(48 ± 5) years] was significantly younger than INSIP-2 group [(52 ± 11) years] and the UIP group [(57 ± 14) years]. The course of disease in INSIP-1 group [(60 ± 28) months] was longer than that in INSIP-2 group [(48 ± 33) months] and that in the UIP group [(44 ± 23) months], but the differences were not statistically significant (F = 1.22, all P > 0.05). The efficacy rate of glucocorticoid treatment in INSIP-1 group (9/9) was higher than that in INSIP-2 group (11/20) and that in the UIP group (2/30), the differences being statistically significant (all P < 0.05). The follow-up period for INSIP-1 group [(56 ± 27) months] was significantly longer than for INSIP-2 group [(23 ± 18) months] and for the UIP group [(25 ± 17) months], and the rate of significant improvement (6/9) was higher than that of the INSIP-2 group (9/20) and the UIP group (0/30), the differences being statistically significant (F = 9.224, all P < 0.05). The mortality of INSIP-1 group (0/9) was lower than that in INSIP-2 group (4/20) and the UIP group (16/30), the difference being statistically significant (exact probability value 0.000 - 0.005, P < 0.05). CONCLUSIONS: The degree of fibrosis of INSIP is closely correlated with the effect of glucocorticoid therapy and prognosis. The cellular type has a favorable reaction to glucocorticoid therapy and a better prognosis as compared to the fibrotic type.

Autocrine BMP4 signaling involves effect of cholesterol myristate on proliferation of mesenchymal stem cells.

We recently identified that cholesterol myristate in traditional Chinese medicine (TCM) is the active compound that increases proliferation of mesenchymal stem cell (MSCs). The present study is further to determine what signal pathway involves in effect of cholesterol myristate. Reverse transcription-PCR, Western blot and ELISA analysis show that cholesterol myristate increases the release of bone morphogenetic protein 4 (BMP4) from MSCs and the expression in the intracellular levels of BMP4 in a time- and dose dependent manner. However, structurally related steroids such as cholesterol and cholesten presented in TCM, both lack of the myristate, did not affect the secretion and expression of BMP4 on MSCs. These finds suggest that myristate is essential for the effects of cholesterol myristate. Furthermore, cholesterol myristate significantly increase BMPRIB levels of MSCs and the number of BMPRIB positive cells in a time- and dose dependent manner, but not BMPR IA or BMPR II. Our results indicate that action of cholesterol myristate may activate the BMP4-BMPRIB autocrine. Moreover, a blocking antibody against BMP4 or the BMP4 antagonist, noggin, partially reduced the effects of cholesterol myristate on MSCs proliferation. Thus, this study is to provide evidence that autocrine BMP4 signaling involves effect of cholesterol myristate on MSCs proliferation.

[Idiopathic lymphoid interstitial pneumonia: a report of 3 cases and literature review].

OBJECTIVE: To analyze the clinical, radiological and pathological characteristics of idiopathic lymphoid interstitial pneumonia (idiopathic LIP) and to discuss its diagnosis, treatment and prognosis. METHODS: Respiratory physicians, pathologists and radiologists together retrospectively analyzed the clinical, chest roentgenogram, computerized tomography, pathological, diagnostic and therapeutic data of 3 patients with idiopathic LIP confirmed by lung biopsy, and reviewed the relevant literatures. RESULTS: The major symptoms of the 3 cases of idiopathic LIP were progressive dyspnea and dry cough. Higher levels of gamma-globulins in serum were found in all the cases. The characteristic radiographic manifestations were bilateral diffuse nodules and cysts. The pathologic feature was diffuse interstitial inflammation with polyclonal lymphocytes infiltration, especially with plasma lymphocytes. Corticosteroids and cytotoxic agents were used and good response to therapy was observed in the cases. CONCLUSIONS: Idiopathic LIP has some characteristics on the clinical, radiological and pathological features, but the best diagnostic method depends on a clinical-radiological-pathological approach. The disease usually shows good response to combinative therapy of corticosteroids and cytotoxic agents.

[Diffuse myxoid malignant fibrous histiocytoma of the pleura: a case report and review of the literature].

OBJECTIVE: To study the clinicopathologic characteristics of diffuse myxoid malignant fibrous histiocytoma (MFH) of pleura. METHODS: The clinical and pathological features of a patient with diffuse myxoid MFH of pleura from this hospital were analyzed, and the reported literature of 7 patients with MFH of pleura were reviewed. RESULTS: The patient was a 65-year-old male. The primary manifestations were cough, chest pain, breathlessness, and a great quantity of bloody pleural effusion. Chest CT scan showed diffuse masses on the visceral and parietal pleura. Exploratory thoracotomy exposed diffuse translucent gray-white masses on the surface of visceral and parietal pleura, with diameters of 1 - 8 cm. Microscopic findings showed that diffuse spindle-shaped and pleomorphic tumour cells were within the myxoid stroma. Tumor cells were positive for vimentin, CD(68), lysozyme, and negative for CK, EMA, and desmin. The patient died of obstruction of superior vena cava, and circulatory failure at 24th postoperative day. Different from cases with single or multiple MFH of pleura reported in the literature, this case of diffuse myxoid MFH had an abrupt onset, and progressed aggressively. CONCLUSION: Diffuse myxoid MFH of pleura is a very rare high-grade malignant tumor with very poor prognosis.

[Clinical analysis of 25 cases of biopsy-proven cryptogenic organizing pneumonia].

OBJECTIVE: To analyze the clinical, radiological and pathological features, diagnosis and response to therapy as well as prognosis of 25 cases of cryptogenic organizing pneumonia (COP). METHODS: Twenty-five subjects with COP confirmed by lung biopsy in Shanghai Pulmonary Hospital from January of 2000 to April of 2006 were retrospectively reviewed. Secondary reaction to infections, drugs, radiation, connective tissue diseases and various noxious agents were excluded. Their clinical-pathological characteristics, radiological features, response to treatment, relapse, survival were obtained from medical records and a follow-up patient questionnaire. RESULTS: There were 6 males and 19 females, with a mean age of 56 years (range 40 - 73 years). The presentations included cough (25/25), clear sputum (21/25), dyspnea (17/25), hemoptysis (5/25), fever and sweats (3/25), and "Velcro" crackles (18/25). Four of them were smokers, 11 had allergic reaction to some drugs, and 11 had some industrious dust inhalation. In 23 cases the specimens were obtained by video-assisted thoracoscopy and 2 cases by transbronchial lung biopsy. Bilateral lung involvement was present in 23 cases and all of them had at least two different radiological manifestations. Twenty-four cases showed a sub-pleural distribution. Bilateral patchy alveolar and ground glass involvement were found in 8 cases, airspace consolidation in 8 cases, mass in 11 cases, irregular lines in 10 cases, small nodules (<10 mm) in 4 cases. Two patients received operation. Corticosteroid therapy was administered to 23 patients. Seventeen cases were cured, but 8 of them relapsed after stopping (n = 2) and tapering (n = 6, when prednisone less than 5 - 10 mg/d) of corticosteroids within one to two years of therapy. CONCLUSIONS: COP is not very rare in China. The clinical-radiological-pathological diagnosis (CRP) is the most important diagnostic method. Corticosteroid is the first choice for COP therapy. The prognosis of COP is good if therapy is started in time, but relapse is common.

[The clinicopathological features and surgical treatment of solitary fibrous tumor of the pleura].

OBJECTIVE: To investigate the clinicopathological features and surgical treatment of solitary fibrous tumor of the pleura (SFTP). METHODS: The clinical, radiological and pathological features of 11 cases of SFTP were analyzed. The detection of immunohistochemistry, surgical treatment and follow-up data were also reviewed. RESULTS: The group consisted of 6 males and 5 females, age ranging from 35 to 73 years (mean 53.7 years). Clinical findings were mainly cough, chest pain and other local symptoms. Five cases had pleural effusion. Surgical excision was performed by thoracotomy in 6 patients, by video-assisted thoracoscopic surgery (VATS) in 3 patients, and by VATS plus a small thoracotomy in 2 patients. Eight cases received tumor resection by wedge resection of the lung, and the other 3 patients by middle and lower lobe resection, lower lobe resection and chest wall tumor resection, respectively. Nine tumors were located in the visceral pleura, 1 tumor below the visceral pleura, and 1 tumor in parietal pleura. Histopathologically, SFTP consisted mainly of spindle-shaped cells which were diffusely distributed. There were abundant gross collagenous fibers and thick-walled blood vessels around tumor cells. Malignant tumors displayed high cell density, nuclear atypia and patho-karyokinesis. The tumor cells were stained with vimentin (100%, 11/11), CD(34) (100%, 11/11), CD(99) (63.6%, 7/11), and bcl-2 (63.6%, 7/11), but no expression of CD(31). Of the 11 cases, 9 were histologically diagnosed as benign SFTP, 1 as cell-abundant type of SFTP, and another as malignant SFTP. One patient died of multiple organ failure after surgery. The mean duration of hospital stay after surgery was 11 days. Follow-up visit results were available for 10 cases. The follow-up time ranged from 12 to 48 months and all patients remained well without recurrence or metastasis. CONCLUSIONS: SFTP should be considered when chest X-ray and CT showed single nodule or mass in the pleura. The diagnosis is based on examination of histopathology and immunohistochemistry. For a tumor with a diameter les than 5 cm, resection by VATS or VATS plus small thoracotomy is a good surgical procedure.