Two cases of spindle cell tumors with S100 and CD34 co-expression showing novel RAF1 fusions.

BACKGROUND: Recently, a novel group of CD34 and S100 co-expression spindle cell tumors with distinctive stromal and perivascular hyalinization harboring recurrent gene fusions involving RET, RAF1, BRAF, and NTRK1/2 gene has been identified. CASE PRESENTATION: In this study, we reported two Chinese male patients with soft tissue tumors presenting in the right knee joint and the left thigh, respectively. For both patients, the tumors were completely excised with clear margin. Microscopically, case 1showed morphological overlap with neurofibroma, and case 2 showed overlap with lipomatous solitary fibrous tumor. Both tumors showed co-expression of S100 and CD34, and absence of SOX10. Genomic profiling with DNA-based next-generation sequencing (NGS) assay was performed and revealed KIF5B-RAF1 (K16:R8) and TLN2-RAF1 (T54:R8) rearrangements. RNA-based NGS and RT-PCR were performed to confirm the gene fusion. CONCLUSIONS: Though systemic therapy was not indicated in these two patients, identification of targetable kinase fusions may help to refine tumors with an ambiguous immunoprofile, and provides suggestions for targeted therapy in rare aggressive cases.

Excessive proliferation and apoptosis of parathyroid cells contribute to primary hyperparathyroidism in rabbit model.

To explore the molecular pathogenesis of primary hyperparathyroidism (PHPT), we investigated the proliferation and apoptosis of parathyroid cells in a rabbit model of diet-induced PHPT. A total of 120 adult Chinese rabbits were randomly divided into normal diet (Ca:P, 1:0.7) group (control group) or a high-phosphate diet (Ca:P, 1:7) group (experimental group). The thyroid and parathyroid complexes were harvested for 1-month interval from month 1 to month 6. The expression of proliferation markers, including proliferating cell nuclear antigen (PCNA) and cyclin-D1, and B cell lymphoma-2 (Bcl-2), were evaluated by immunohistochemistry in thyroid and parathyroid tissues. Apoptosis was quantified by DNA-fragment terminal labeling. Our results demonstrated that parathyroid cells in the experimental group started proliferating from the end of the 2nd month, the expression of PCNA, Bcl-2, and cyclin-D1 were significantly higher in the PHPT group than those of the control group (p<0.05). Furthermore, the apoptosis index (AI) was positively correlated with the glandular cell count and expression of PCNA in the 6th month in the PHPT group. Overall, our results suggested that excessive proliferation and apoptosis of parathyroid cells may contribute to the pathogenesis of PHPT through PCNA-related, Bcl-2-related, and cyclin-D1-related pathways.

Dedifferentiated Central Chondrosarcoma: A Clinical, Histopathological, and Immunohistochemical Analysis of 57 Cases.

Dedifferentiated central chondrosarcoma (DCCS) is a rare cartilage tumor with invasive biological behavior and a poor prognosis. To better understand the morphological characteristics of this type of tumor and its internal mechanism of dedifferentiation, we retrospectively analyzed 57 cases of DCCS. A total of 29 female and 28 male patients were included, ranging in age from 20 to 76 years, with a median age of 54 years. Fifty-seven cases of DCCS occurred in the pelvis (n = 29), femur (n = 17), scapula (n = 4), tibia (n = 2), humerus (n = 2), metatarsals (n = 1), fibula (n = 1), and radius (n = 1). Radiologically, DCCS had two different appearances on imaging, with an area showing calcifications of the cartilage forming the tumor juxtaposed to a lytic area with a highly aggressive, non-cartilaginous component. Histopathologically, the distinctive morphological features consisted of two kinds of defined components: a well-differentiated cartilaginous tumor and non-cartilaginous sarcoma. The cartilaginous components included grade 1 (n = 38; 66.7%) and grade 2 (n = 19; 33.3%) cartilage. The sarcoma components included those of osteosarcoma (n = 29; 50.9%), undifferentiated pleomorphic sarcoma (n = 20; 35.1%), rhabdomyosarcoma (n = 3; 5.2%), fibrosarcoma (n = 2; 3.5%), spindle cell sarcoma (n = 2; 3.5%) and angiosarcoma (n = 1; 1.8%). Immunohistochemistry showed that the expression of p53 and RB in the sarcoma components was significantly higher than that in the cartilaginous components, suggesting that these factors play roles in the dedifferentiation process of chondrosarcoma. DCCS is a highly malignant tumor with a poor prognosis. Except for the patients who were lost to follow-up, most of our patients died.

Survivin Promotes Piperlongumine Resistance in Ovarian Cancer.

Ovarian cancer is one of the most fatal female malignancies while targeting apoptosis is critical for improving ovarian cancer patients' lives. Survivin is regarded as the most robust anti-apoptosis protein, and its overexpression in ovarian cancer is related to poor survival and apoptosis resistance. Piperlongumine (PL) extracted from peppers is defined as an active alkaloid/amide and exhibits a broad spectrum of antitumor effects. Here, we demonstrate that PL induces the rapid depletion of survivin protein levels via reactive oxygen species (ROS)-mediated proteasome-dependent pathway in vitro, while exerting a remarkable inhibitory influence on the proliferation of ovarian cancer cells. Overexpression of survivin raises the survival rate of ovarian cancer cells to PL. Moreover, PL inhibits ovarian cancer cells xenograft tumor growth and downregulates survivin in vivo. Our findings reveal a previously unrecognized mechanism of PL in suppressing survivin expression as well as survivin promotes piperlongumine resistance in ovarian cancer and suggest that ROS-mediated proteasome-dependent pathway can be exploited to overcome apoptosis resistance triggered by aberrant expression of survivin.

Diagnosis and Differential Diagnosis of Desmoplastic Fibroblastoma by Clinical, Radiological, and Histopathological Analyses.

BACKGROUND: Desmoplastic fibroblastoma (collagenous fibroma) is an uncommon benign soft-tissue tumor, rarely involving bone. It shares some overlapping features with other infiltrate tumors, such as desmoid-type fibromatosis, neurofibroma, and low-grade fibromyxoid sarcoma. The misdiagnosis may cause unnecessary surgical overtreatment, especially for those involving bone. In order to deepen the understanding of the diagnosis and differential diagnosis of desmoplastic fibroblastoma, we planned to analyze the clinical, radiological, and histopathological features and the outcome of desmoplastic fibroblastoma on the basis of case analysis and literature review. METHODS: Sixteen cases were retrieved from the surgical pathology records from May 2011 to April 2016 in the Department of Pathology in Beijing Jishuitan Hospital. Formalin-fixed, paraffin-embedded specimens of 16 cases of desmoplastic fibroblastoma were collected. Hematoxylin and eosin stain and immunohistochemistry were used to observe the histological features of desmoplastic fibroblastoma of soft tissue and bone. The images for diagnosis obtained from the ultrasonic examination, X-ray, magnetic resonance imaging, and computed tomography were used to observe the radiological features. Related literatures were retrieved from the PubMed and CNKI databases. RESULTS: Sixteen cases of desmoplastic fibroblastoma of soft tissue were located in the hand (n = 7), foot (n = 4), upper arm (n = 1), shoulder (n = 1), forearm (n = 2), and one case occurred in the proximal femur. Age ranged from 32 to 82 years (median age: 58 years). There were six females and ten males. Histologically, the lesions of soft tissue appeared as well-circumscribed masses with abundant collagenous matrix and low vascularity. Tumor cells were stellate- or spindle-shaped and uniformly distributed within the extracellular matrix. In five cases, the desmoplastic fibroblastoma were found to have infiltrated into the skeletal muscle tissue. In one case of desmoplastic fibroblastoma of bone, radiographs revealed osteolytically well-defined lesion. Immunohistochemistry stain showed that vimentin and smooth muscle actin were positive in all cases of desmoplastic fibroblastoma. CONCLUSIONS: Desmoplastic fibroblastoma (collagenous fibroma) has prominent clinical, histopathological, and radiological features. Before the differential diagnosis from other tumors is obtained by thorough analysis and comparison of the similar and different characteristics, the appropriate surgical management and accurate prognosis evaluation could not be delivered to the patient.

Evaluation of retinal nerve fiber layer thickness in patients with obstructive sleep apnea-hypopnea syndrome / 国际眼科杂志(Guoji Yanke Zazhi)

AIM:To compare the retinal nerve fiber layer (RNFL) thickness in patients with different severities of obstructive sleep apnea-hypopnea syndrome (OSAHS) versus normal controls.METHODS:This was an observational case-control study.Totally 79 patients presenting with snoring and daytime sleepiness who underwent overnight polysomnography (PSG) to determine OSAHS severity were recruited,and subsequently referred for ophthalmologic evaluation.The data including intraocular pressure (lOP),RNFL thickness etc was collected.RESULTS:A total of 73 subjects were recruited,including 53 patients with OSAHS and 20 normal control subjects.lOP was significantly higher for the moderate and severe OSAHS group than for the control group (both P＜0.05).The RNFL in the average and superior quadrants were significantly thinner for the moderate and severe OSAHS group than for the control group (both P＜0.05).AHI was negatively correlated with RNFL thickness in the superior quadrants,and with average RNFL thickness (r=-0.316,-0.205;both P＜0.01).Negative correlations were also identified between RNFL thickness in the superior quadrants and the ODI (r=-0.24,P =0.005).Positive correlations were identified between SaO2 on PSG and RNFL thickness in the superior quadrants (r=0.277,P=0.001).CONCLUSION:RNFL of the patients with OSAHS is thinned.Furthermore,it is observed that the greater severity of OSAHS,the greater the loss of RNFL.Among the four quadrants observed,the most affected quadrant was the superior quadrant.

[High Throughput Screening Analysis of Preservatives and Sweeteners in Carbonated Beverages Based on Improved Standard Addition Method].

Simulated water samples of 3 kinds of preservatives and 4 kinds of sweeteners were formulated by using orthogonal design. Kernel independent component analysis (KICA) was used to process the UV spectra of the simulated water samples and the beverages added different amounts of the additive standards, then the independent components (ICs), i. e. the UV spectral profiles of the additives, and the ICs' coefficient matrices were used to establish UV-KICA-SVR prediction model of the simulated preservatives and sweeteners solutions using support vector regression (SVR) analysis. The standards added beverages samples were obtained by adding different amounts level of additives in carbonated beverages, their UV spectra were processed by KICA, then IC information represented to the additives and other sample matrix were obtained, and the sample background can be deducted by removing the corresponding IC, other ICs' coefficient matrices were used to estimate the amounts of the additives in the standard added beverage samples based on the UV-KICA-SVR model, while the intercept of linear regression equation of predicted amounts and the added amounts in the standard added samples is the additive content in the raw beverage sample. By utilization of chemometric "blind source separation" method for extracting IC information of the tested additives in the beverage and other sample matrix, and using SVR regression modeling to improve the traditional standard addition method, a new method was proposed for the screening of the preservatives and sweeteners in carbonated beverages. The proposed UV-KICA-SVR method can be used to determine 3 kinds of preservatives and 4 kinds of sweetener in the carbonate beverages with the limit of detection (LOD) are located with the range 0.2-1.0 mg · L⁻¹, which are comparable to that of the traditional high performance liquid chromatographic (HPLC) method.

Cyclin-dependent kinase inhibitor dinaciclib potently synergizes with cisplatin in preclinical models of ovarian cancer.

Ovarian cancer is one of the most lethal of woman cancers, and its clinical therapeutic outcome currently is unsatisfied. Dinaciclib, a novel small molecule inhibitor of CDK1, CDK2, CDK5 and CDK9, is assessed in clinical trials for the treatment of several types of cancers. In this study, we investigated the anticancer effects and mechanisms of dinaciclib alone or combined with cisplatin in ovarian cancer. Dinaciclib alone actively induced cell growth inhibition, cell cycle arrest and apoptosis with the increased intracellular ROS levels, which were accompanied by obvious alterations of related proteins such as CDKs, Cyclins, Mcl-1, XIAP and survivin. Pretreatment with N-acety-L-cysteine significantly blocked ROS generation but only partially rescued apoptosis triggered by dinaciclib. Moreover, the combination of dinaciclib with cisplatin synergistically promoted cell cycle arrest and apoptosis, and inhibited the subcutaneous xenograft growth of ovarian cancer in nude mice. Altogether, dinaciclib potently synergizes with cisplatin in preclinical models of ovarian cancer, indicating this beneficial combinational therapy may be a promising strategy for treatment of ovarian cancer.

Piperlongumine induces apoptosis and synergizes with cisplatin or paclitaxel in human ovarian cancer cells.

Piperlongumine (PL), a natural alkaloid from Piper longum L., possesses the highly selective and effective anticancer property. However, the effect of PL on ovarian cancer cells is still unknown. In this study, we firstly demonstrate that PL selectively inhibited cell growth of human ovarian cancer cells. Furthermore, PL notably induced cell apoptosis, G2/M phase arrest, and accumulation of the intracellular reactive oxidative species (ROS) in a dose- and time-dependent manner. Pretreatment with antioxidant N-acety-L-cysteine could totally reverse the PL-induced ROS accumulation and cell apoptosis. In addition, low dose of PL/cisplatin or paclitaxel combination therapies had a synergistic antigrowth effect on human ovarian cancer cells. Collectively, our study provides new therapeutic potential of PL on human ovarian cancer.

Triterpenoids as reversal agents for anticancer drug resistance treatment.

Overexpression of ATP-binding cassette (ABC) transporters in cancer cells results in multidrug resistance (MDR), which is one of the major obstacles in the treatment of cancer patients. None of the strategies to overcome MDR has been successfully applied in the clinic until now. Plenty of evidence shows that some triterpenoids function as reversal agents of MDR for anticancer drug resistance treatment. Here, we review the latest findings of reversing cancer MDR with triterpenoids. Findings are summarized showing that triterpenoids are MDR modulators and potential chemosensitizers. Finally, we contemplate future prospects of modulating MDR in the clinic.

Extracellular ATP enhances in vitro invasion of prostate cancer cells by activating Rho GTPase and upregulating MMPs expression.

We previously found that in addition to anti-proliferation function, extracellular ATP had a pro-invasion effect on prostate cancer cells, and probably serves as an important regulator of invasion in local microenvironment. However, the underlying mechanism remains unclear. In this study, we demonstrated that ATP increased the motility of prostate cancer cells, and promoted formation of lamellipodia and filopodia. We also found that ATP induced activation of Rac1 and Cdc42, and promoted expression of MMP-3 and MMP-13. These data suggest that extracellular ATP enhances the invasion of prostate cancer cells by activating Rho GTPases Rac1 and Cdc42 and upregulating MMPs expression.

[Giant cell tumor of bone and malignancies in giant cell tumor: a clinicopathologic analysis].

OBJECTIVE: To study the clinicopathologic features, diagnosis and differential diagnosis of malignancies in giant cell tumor (MGCT). METHODS: The clinicopathologic features of 13 cases of MGCT were retrospectively reviewed. RESULTS: Thirteen cases of MGCT were found amongst a total of 603 cases of giant cell tumor encountered. Six of the 13 cases represented concurrent malignancy in giant cell tumor while the remaining 7 cases was malignant transformation in recurrent giant cell tumor. The age of the patients ranged from 21 to 71 years (mean age = 39.5 years) in the first group and from 27 to 52 years (mean age = 36.7 years) in the second group. In concurrent MGCT, a high-grade sarcoma component was present in conjunction with the giant cell tumor component. In malignant transformation of recurrent giant cell tumor, the original tumor was giant cell tumor and the recurrence showed features reminiscent of malignant fibrous histiocytoma. CONCLUSIONS: The diagnosis of malignancies in giant cell tumor requires correlation of clinical, radiologic and pathologic features. The entities need to be distinguished from other giant cell-rich tumors including primary malignant fibrous histiocytoma and giant cell osteosarcoma.

[Myositis ossificans: a clinicopathologic analysis of 15 cases].

OBJECTIVE: To study the clinicopathologic features, diagnosis and differential diagnosis of myositis ossificans (MO). METHODS: The clinical features, radiologic results and pathologic findings of 15 cases of MO (including biopsy and surgical specimens) were analyzed. The hematoxylin and eosin sections were reviewed under light microscope. Immunohistochemical staining for S-100 protein, vimentin, desmin, actin and osteonectin was performed. RESULTS: The age of the patients ranged from 12 to 46 years. The male-to-female ratio was 11:4. Thirteen cases were located in the parosteum of long bone or subperiosteal soft tissue. The remaining two cases occurred in iliac region and palm, respectively. Five patients had history of injury, while 2 patients had operation before. Four patients had no history of trauma and the remaining one had unknown clinical history. Histologically, zonation pattern was not conspicuous in 10 biopsy cases and 8 corresponding surgical specimens. On the other hand, zonation pattern was observed in 5 biopsy cases and 7 corresponding surgical specimens. Follow up revealed relapses in two patients. Immunohistochemical study showed various degree of positivity for vimentin, desmin, actin and osteonectin. S-100 protein was focally positive in 2 of the cases. The Ki-67 index varied from 1% to 10%. CONCLUSION: Correct diagnosis of MO relies on correlation of clinical features, radiologic examination and pathologic findings.

Basic FGF augments hypoxia induced HIF-1-alpha expression and VEGF release in T47D breast cancer cells.

AIM: Both hypoxia inducible factor 1 (HIF-1) and basic fibroblast growth factor (bFGF) play important roles in tumour angiogenesis. This study was designed to clarify the cooperative effect of these two mediators in induction of vascular endothelial cell growth factor (VEGF) release from breast cancer and probe possible mechanisms involved. METHODS: Release of VEGF from a breast cancer cell line (T47D) was quantitated by enzyme linked immunosorbent assay (ELISA). Expression of HIF-1 and ERK was assayed using Western blotting. Transient transfection and dual luciferase reporter assay were used to study HIF-1 transactivity. RESULTS: The data showed that hypoxia induced the expression of HIF-1alpha protein, the transactivity of HIF-1 and the release of VEGF. bFGF further augmented these hypoxic inductions. The PI3K pathway was required for these processes as demonstrated by application of PI3Kinase inhibitor (LY294002) or mutant construct transfections. In contrast, the MEK1 inhibitor PD98059 showed no effect on either activation of HIF-1 or VEGF release, which is in agreement with our finding that ERK1/2 was not activated by hypoxia. Under hypoxic conditions, bFGF activated the MEK1/ERK pathway. PD98059 blocked the activation of ERK1/2 and suppressed bFGF-induced HIF-1 transactivity, yet the protein expression of HIF-1alpha or VEGF release was not affected by PD98059. CONCLUSION: bFGF augments hypoxia induced VEGF release mainly through the PI3K pathway and partly depending on HIF-1 activity. Elucidation of this mechanism may provide a new target for anti-angiogenesis in cancer therapy.

[P2Y purinergic receptor activated PI-3K/Akt signaling pathway in regulation of growth and invasion of prostatic cancer].

OBJECTIVE: To investigate P2Y purinergic receptor activated PI-3K/Akt signaling pathway in the regulation of growth and invasion of prostate cancer in vitro. METHODS: Western blot was used to detect phosphorylation of Akt (a downstream target molecule of PI-3K) by P2Y receptor agonist in 1E8 cells (a highly metastatic subclone derived from PC-3 prostatic cancer cell line). Cell counts, flow cytometry, Matrigel invasion assay, wound healing assay and gelatin zymography were used to detect changes of biological behaviors of 1E8 cells after P2Y receptor activation. RESULTS: AMP-PNP, one non-hydrolysis ATP analogue and P2Y receptor agonist, induced significant phosphorylation of Akt in a time- and dose-dependent manner in IE8 cells. LY294002, a specific inhibitor of PI-3K, effectively blocked Akt phosphorylation induced by AMP-PNP. Continuous exposure to AMP-PNP induced significant growth inhibition of 1E8 cells (inhibition rate at 50.2% at the 8th day), and this inhibition was mainly due to an arrest at S phase of the cell cycle (the S phase fraction of AMP-PNP treated cells was 22.3% higher than that of the control). Application of LY294002 did not reverse the growth inhibition effect of AMP-PNP. Matrigel invasion assay showed that AMP-PNP stimulation increased invasive ability of 1E8 cells, and this effect was effectively blocked by LY294002. No significant changes in the activation of MMP-2 and MMP-9 were detected by gelatin zymography, although wound healing assay showed 21.2% increase in cell migration after AMP-PNP treatment. CONCLUSIONS: PI-3K/Akt signaling pathway participates in P2Y receptor-stimulated prostate cancer invasion by enhancing cell motility, rather than up-regulating MMP-2 and MMP-9 activities. PI-3K signaling pathway plays an important role in prostate cancer proliferation, but is not involved in P2Y receptor mediated growth inhibition.

In vitro study of HIF-1 activation and VEGF release by bFGF in the T47D breast cancer cell line under normoxic conditions: involvement of PI-3K/Akt and MEK1/ERK pathways.

Hypoxia-inducible factor (HIF) is critical in the modulation of tumour angiogenesis in response to hypoxia. In the present study, the mechanisms underlying basic fibroblast growth factor (bFGF)-induced activation of HIF-1 and the subsequent release of vascular endothelial growth factor (VEGF) in a human breast cancer cell line (T47D) under normoxic conditions were explored. The data show that HIF-1alpha expression is induced by bFGF in a dose- and time-dependent fashion, while increased HIF-1alpha protein expression and transactivity of HIF-1 are due to the phosphorylation of Akt by bFGF, as indicated by application of the phosphatidylinositol 3-kinase (PI-3K) inhibitor LY294002. The data also show that the MEK1 (mitogen-activated protein kinase kinase-1)/ERK (extracellular signal-regulated kinase) pathway is only involved in bFGF-induced transactivity of HIF-1, but not HIF-1alpha expression, indicating roles for both the PI-3K/Akt and the MEK1/ERK pathways in bFGF activity. In addition, the translation inhibitor cycloheximide confirmed that bFGF-induced HIF-1alpha protein expression was due to de novo protein synthesis. In contrast, p38 was not required for the expression of HIF-1alpha or HIF-1 transactivity, although significant phosphorylation of p38 was observed after bFGF treatment. Treatment of the cells with bFGF increased the amount of VEGF release, and this could be suppressed by either PD98059 or LY294002, suggesting the presence of a HIF-1alpha-dependent pathway for bFGF-induced VEGF production. In conclusion, the PI-3K/Akt and MEK1/ERK pathways, in a potentially independent and co-operative fashion, can modulate HIF-1 activation by bFGF. Further studies will pinpoint whether HIF-1 is the transcriptional factor responsible for the increased VEGF production following bFGF treatment of breast tumour cells.

[Melatonin in in vitro apoptosis of H22 hepatocarcinoma cells].

OBJECTIVE: To study the effect of melatonin (MLT) in in vitro apoptosis of hepatocarcinoma cells and its mechanism. METHODS: The apoptotic cells, bcl-2 and bax were detected through immunocytochemical method (ICC) and Tolt-mediated x-duTP nick end labeling (TUNEL). Computer image analysis system was used to quantify the expression of bcl-2 and bax by detecting the absorbance value of positive products. Apoptosis index (AI) was used to quantify the number of apoptotic cells. RESULTS: In vitro, AI increase was both concentration- and time-dependent through TUNEL. During the same duration, AI of medium dose group was higher than that of low dose and control group (P < 0.05); AI of high dose, medium dose and 5-Fu group were higher than those of low dose and control group (P < 0.01), however, there was no significant difference between the low dose and control group (P > 0.05). At the same dose, in high dose, medium dose and 5-Fu group, the change of AI showed significant difference from 24 to 36 hours (P < 0.05). The expression of bcl-2 was down-regulated as the MLT increased, and there was significant difference between the low dose and control group (P < 0.01). But, the expression of bax was up-regulated as the dose of MLT increased, showing significant difference between the high dose and control groups (P < 0.01). As time went on, the expression of bcl-2 was decreased and in every group, with the change in absorbance value of bcl-2 significantly different from 24 to 36 hours (P < 0.05), whereas that of bax remained almost unchanged. The ratio of bax/bcl-2 was increased with the increase in the concentration of MLT. CONCLUSION: Melatonin may induce apoptosis in the hepatocarcinoma cells which is concentration- and time-dependent, in which bcl-2 and bax are involved.