OBJECTIVETo investigate the anti-cancer effects of crude extract from Melia toosendan Sieb. et Zucc and its possible molecular mechanisms in vitro and in vivo.

METHODSTransonic alcohol-chloroform extraction method was used to extract toosendanin from the bark of Melia toosendan Sieb. et Zucc, and the content of toosendanin in the crude extract was measured by high performance liquid chromatography (HPLC). Anti-cancer effects of crude extract from Melia toosendan Sieb. et Zucc were investigated in in vivo and in vitro studies. In the in vitro experiment, human hepatocellular carcinoma cell lines SMMC-7721 and Hep3B were co-incubated with toosendanin crude extract of different concentrations, respectively. In the in vivo experiment, BALB/c mice were subcutaneously inoculated with mouse hepatocellular carcinoma H22 cells and treated with crude extract.

RESULTSHPLC revealed the content of toosendanin was about 15%. Crude extract from Melia toosendan Sieb. et Zucc inhibited cancer cells growth in a dose- and time-dependent manner. The 50% inhibitory concentration (IC50, 72 h) was 0.6 mg/L for SMMC-7721 cells and 0.8 mg/L for Hep3B cells. Both high-dose [0.69 mg/(kg d)] and low-dose [0.138 mg/(kg d)] crude extract could markedly suppress cancer growth, and the inhibition rate was greater than 50%. Hematoxylin and eosin staining showed necrotic area in cancers and transmission electron microscopy displayed necrotic and apoptotic cancer cells with apoptotic bodies. Immunohistochemistry showed that the expression of Bax and Fas increased and the expression of Bcl-2 reduced.

CONCLUSIONSToosendanin extract has potent anti-cancer effects via suppressing proliferation and inducing apoptosis of cancer cells in vivo and in vitro. The mechanism of apoptosis involves in mitochondrial pathway and death receptor pathway.

Animals ; Antineoplastic Agents ; pharmacology ; therapeutic use ; Apoptosis ; drug effects ; Carcinoma, Hepatocellular ; drug therapy ; pathology ; ultrastructure ; Cell Proliferation ; drug effects ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; therapeutic use ; Female ; Immunohistochemistry ; Liver Neoplasms ; drug therapy ; pathology ; ultrastructure ; Male ; Melia ; chemistry ; Mice, Inbred BALB C ; Mitochondria ; drug effects ; metabolism ; Neoplasm Transplantation ; Plant Extracts ; therapeutic use ; Reference Standards ; bcl-2-Associated X Protein ; metabolism ; fas Receptor ; metabolism

Cerebellar Neoplasms ; ultrastructure ; Humans ; Medulloblastoma ; ultrastructure

OBJECTIVETo study the expression of nNOS and ultrastructural changes in the penile tissue of rats with prolactinoma-induced erectile dysfunction (ED).

METHODSWe established the model of prolactinoma in 20 male Westar rats by peritoneal injection of diethylstilbestrol (DES) and treated the control rats with normal saline (n = 10) or sterilized arachis oil (n = 10). After 8 weeks, we performed the apomorphine test and measured the weight of the pituitary gland and the levels of serum prolactin (PRL) and testosterone (T) to confirm the successful construction of the prolactinoma-induced ED model. Then we determined the expression of nNOS in the penile tissue by immunohistochemistry and examined the ultrastructural changes of the penile cavernosum under the transmission electron microscope.

RESULTSThe prolactinoma-induced ED model was successfully established in 15 rats. The weight of the pituitary gland was significantly increased in the rats treated with DES as compared with the normal saline and sterilized arachis oil controls ([46.7 ± 15.5] vs [11.7 ± 2.4] and [12.4 ± 2.3] mg, both P < 0.05). The level of serum PRL was markedly higher while that of T remarkably lower in the former than in the latter two groups ([1,744.9 ± 304.5] vs [11.5 ± 2.4] and [10.6 ± 1.9] ng/ml, both P < 0.0l; [1.54 ± 0.46] vs [3.11 ± 1.08] and [3.04 ± 1.11] ng/ml, both P < 0.05). The rate of penile erection was significantly reduced in the prolactinoma-induced ED model rats in comparison with the normal saline and arachis oil controls (16.7% vs 100% and 87.5%, both P < 0.05), and so was the expression of nNOS in the penile tissue (0.024 ± 0.011 vs 0.066 ± 0.019 and 0.058 ± 0.021, both P < 0.05). Transmission electron microscopy manifested significant ultrastructural changes in the endothelial and smooth muscle cells of the cavernous tissue in the prolactinoma-induced ED models.

CONCLUSIONThe ultrastructural changes of the penile cavernous tissue and the reduced expression of nNOS in penile tissue may be the most important mechanisms of prolactinoma-induced ED in rats.

Animals ; Apomorphine ; Carcinogens ; Diethylstilbestrol ; Erectile Dysfunction ; etiology ; Humans ; Male ; Myocytes, Smooth Muscle ; ultrastructure ; Nitric Oxide Synthase Type I ; metabolism ; Organ Size ; Penile Erection ; Penis ; enzymology ; ultrastructure ; Pituitary Neoplasms ; chemically induced ; complications ; Prolactin ; blood ; Prolactinoma ; chemically induced ; complications ; Rats ; Rats, Wistar ; Testosterone ; blood

Stem cells are known to maintain stemness at least in part through secreted factors that promote stem-like phenotypes in resident cells. Accumulating evidence has clarified that stem cells release nano-vesicles, known as exosomes, which may serve as mediators of cell-to-cell communication and may potentially transmit stem cell phenotypes to recipient cells, facilitating stem cell maintenance, differentiation, self-renewal, and repair. It has become apparent that stem cell-derived exosomes mediate interactions among stromal elements, promote genetic instability in recipient cells, and induce malignant transformation. This review will therefore discuss the potential of stem cell-derived exosomes in the context of stromal remodeling and their ability to generate cancer-initiating cells in a tumor niche by inducing morphologic and functional differentiation of fibroblasts into tumor-initiating fibroblasts. In addition, the immunosuppressive potential of stem cell-derived exosomes in cancer immunotherapy and their prospective applications in cell-free therapies in future translational medicine is discussed.
Apoptosis ; Cell Communication ; Cell Transformation, Neoplastic ; Disease Progression ; Exosomes ; physiology ; Humans ; Immunotherapy ; methods ; Mesenchymal Stromal Cells ; physiology ; Neoplasms ; blood supply ; pathology ; therapy ; Neoplastic Stem Cells ; ultrastructure ; Neovascularization, Pathologic ; pathology ; Organelle Biogenesis ; Tumor Microenvironment

OBJECTIVE: To compare the diagnostic performance of six different approaches for assessing myometrial infiltration using ultrasound in women with carcinoma of the corpus uteri. METHODS: Myometrial infiltration was assessed by two-dimensional (2D) transvaginal or transrectal ultrasound in 169 consecutive women with well (G1) or moderately (G2) differentiated endometrioid type endometrial carcinoma. In 74 of these women three-dimensional (3D) ultrasound was also performed. Six different techniques for myometrial infiltration assessment were evaluated. The impression of examiner and Karlsson's criteria were assessed prospectively. Endometrial thickness, tumor/uterine 3D volume ratio, tumor distance to myometrial serosa (TDS), and van Holsbeke's subjective model were assessed retrospectively. All subjects underwent surgical staging within 1 week after ultrasound evaluation. Definitive histopathological data regarding myometrial infiltration was used as gold standard. Sensitivity and specificity for all approaches were calculated and compared using McNemar test. RESULTS: The impression of examiner and subjective model performed similarly (sensitivity 79.5% and 80.5%, respectively; specificity 89.6% and 90.3%, respectively). Both methods had significantly better sensitivity than Karlsson's criteria (sensitivity 31.8%, p<0.05) and endometrial thickness (sensitivity 47.7%, p<0.05), and better specificity than tumor/uterine volume ratio (specificity 28.3%, p<0.05) and TDS (specificity 41.5%, p<0.05). CONCLUSION: Subjective impression seems to be the best approach for assessing myometrial infiltration in G1 or G2 endometrioid type endometrial cancer by transvaginal or transrectal ultrasound. The use of mathematical models and other objective 2D and 3D measurement techniques do not improve diagnostic performance.
Adult ; Aged ; Aged, 80 and over ; Carcinoma, Endometrioid/pathology/*ultrasonography ; Endometrial Neoplasms/pathology/*ultrastructure ; Female ; Humans ; Imaging, Three-Dimensional ; Middle Aged ; Models, Theoretical ; Myometrium/pathology/ultrasonography ; Neoplasm Invasiveness/pathology/*ultrasonography ; Prospective Studies ; Retrospective Studies ; Tumor Burden

OBJECTIVETo evaluate the accuracy of endorectal ultrasound (ERUS) in predicting the circumferential resection margin (CRM) and maximum tumor thickness (MTT) of in T3 rectal cancer.

METHODSClinicl data of 53 patients with pT3 rectal cancer admitted to the Department of General Surgery in the Peking Union Medical College Hospital from June 2011 to January 2014 were retrospectively analyzed. CRM and MTT measured by ERUS were compared with corresponding pathologic measurements to assess the accuracy of ERUS diagnosis.

RESULTSERUS correctly predicted CRM status in 52 patients (98.1%, 52/53), whose sensitivity was 100%, specificity was 97.8%, positive predictive value was 85.7%, and negative predictive value was 100%. ERUS correctly predicted MTT status in 51 patients (96.2%, 51/53), whose sensitivity was 100%, specificity was 95.5%, positive predictive value was 66.6%, and negative predictive value was 100%. In the Bland and Altman plot, the agreement between ERUS and pathology was good.

CONCLUSIONEndorectal ultrasonography can accurately diagnose CRM and MTT, which can satisfy the clinical need for preoperative staging of rectal cancer.

Colectomy ; Humans ; Neoplasm Staging ; Peritoneum ; Rectal Neoplasms ; ultrastructure ; Retrospective Studies

Colloidal particle size is an important characteristic that allows mapping sentinel nodes in lymphoscintigraphy. This investigation aimed to introduce different ways of making a 99mTc-tin colloid with a size of tens of nanometers. All agents, tin fluoride, sodium fluoride, poloxamer-188, and polyvinylpyrrolidone (PVP), were mixed and labeled with 99mTc. Either phosphate or sodium bicarbonate buffers were used to adjust the pH levels. When the buffers were added, the size of the colloids increased. However, as the PVP continued to increase, the size of the colloids was controlled to within tens of nanometers. In all samples, phosphate buffer added PVP (30 mg) stabilized tin colloid (99mTc-PPTC-30) and sodium bicarbonate solution added PVP (50 mg) stabilized tin colloid (99mTc-BPTC-50) were chosen for in vitro and in vivo studies. 99mTc-BPTC-50 (<20 nm) was primarily located in bone marrow and was then secreted through the kidneys, and 99mTc-PPTC-30 (>100 nm) mainly accumulated in the liver. When a rabbit was given a toe injection, the node uptake of 99mTc-PPTC-30 decreased over time, while 99mTc-BPTC-50 increased. Therefore, 99mTc-BPTC-50 could be a good candidate radiopharmaceutical for sentinel node detection. The significance of this study is that nano-sized tin colloid can be made very easily and quickly by PVP.
Animals ; Buffers ; Cell Line, Tumor ; Humans ; Lymph Nodes/*radionuclide imaging ; Lymphatic Metastasis ; Metal Nanoparticles/chemistry/ultrastructure ; Mice ; Neoplasms, Experimental/*radionuclide imaging ; Particle Size ; Povidone/*chemistry ; Rabbits ; Radiopharmaceuticals/*chemical synthesis ; Reproducibility of Results ; Sensitivity and Specificity ; Technetium Compounds/*chemistry ; Tin/*chemistry ; Tin Compounds/*chemistry

OBJECTIVE: To provide a reference about choosing the methods of isolating exosomes derived from tumor cells including laryngocarcinima Hep-2 cells by comparing advantages and defects of two methods of isolation and extraction exosomes. METHOD: Previously, laryngocarcinoma Hep-2 cells were cultivated massively, then the cells were processed with hot shock in 42 degrees C for 1 h. Sucrose density gradient centrifugation ultrafiltration (method 1): cells culture supernatant 90 ml was gathered, the supernatant was clarified through a 3/0.8 μm small filter to remove impurities and fragments which in larger diameter. Then the filtering fluid was concentrated and purified through sucrose density gradient centrifugation and ultrafiltration, the concentrated fluid was obtained. Exosome Isolation Kit (method 2): cells culture supernatant 4 ml was gathered, the solutions of the kit were added into the supernatant in proper sequence, then filtered by the special column, the concentrated fluid was obtained. Both products are observed by high resolution transmission electron microscopy. RESULT: Both methods could isolate and extract exosomes feasibly. In single high power view of transmission electron microscopy, exosomes of method 1 disperse better, but lower density, and more impurity in background, exosomes of method 2 arrange closer, higher density, and less impurity. CONCLUSION Exosome isolation Kit require less supernatant, cost less time, process procedure briefly, harvest higher yield. It may become a new option of isolating exosomes derived from Laryngocarcinoma Hep-2 cells.
Cell Line, Tumor ; Exosomes ; ultrastructure ; Humans ; Laryngeal Neoplasms ; pathology ; Microscopy, Electron, Transmission

OBJECTIVE: To observe the micromorphological changes of ultrastructure, apoptosis-related proteins expression and tumor cell apoptosis after ablation with the high-intensity focused ultrasound (HIFU), and to explore the mechanisms responsible for the thermal and non-thermal effect.
 METHODS: Forty rabbits with hepatic VX2 tumors were randomly divided into a thermal group (n=20) and a non-thermal group (n=20), and were subjected to HIFU ablation with thermal or non-thermal condition, respectively. Five animals in each group were sacrificed on the 1st, 3rd, 7th or 14th day after the ablation. The changes of ultrastructure, apoptosis-related proteins expression and tumor cell apoptosis were detected.
 RESULTS: The results of transmission electron microscope (TEM) revealed more severe injury on tissue and cells in the non-thermal group than that in the thermal group. The changes of apoptosis-related proteins expression and tumor cell apoptosis in transient zone were significantly different in comparison with that in the ablated area or peripheral area between the two groups. The expression of vascular endothelial growth factor (VEGF) was at low level on the 1st and 3rd day and elevated gradually on the 7th and 14th day, with no significant difference (all P>0.05). The expression of caspase-3 reached peak on the 3rd day and decreased on the 7th and 14th day. It was significantly higher in the non-thermal group than that in the thermal group on the 3rd and 7th day (all P<0.05). The expression of NF-κB was elevated from the 3rd day and reached peak on the 7th day while decreased on the 14th day. There was no significant difference at every time point between the 2 groups (all P>0.05). The apoptosis index in the non-thermal group and the thermal group on the 3rd and 7th day were (28.60±1.14)% vs (21.80±1.92)% and (21.00±1.58)% vs (14.80±1.48)%, respectively. It was higher in the non-thermal group than that in the thermal group (both P<0.01).
 CONCLUSION Both the thermal and the non-thermal effect of HIFU can induce apoptosis in transient zone, but the latter have a stronger effect.
Animals ; Apoptosis ; Caspase 3 ; metabolism ; High-Intensity Focused Ultrasound Ablation ; Liver Neoplasms ; pathology ; ultrastructure ; NF-kappa B ; metabolism ; Neoplasms, Experimental ; pathology ; ultrastructure ; Rabbits ; Vascular Endothelial Growth Factor A ; metabolism

OBJECTIVETo study the clinicopathological features, differential diagnosis and prognosis of clear cell papillary renal cell carcinoma (CCPRCC).

METHODSThe histological, immunohistochemical, and molecular features were studied in 11 cases and follow-up data were also analyzed.

RESULTSThere were a total of 3 females and 8 males. The age of patients were ranged from 33 to 72 years(mean age 52.5 years). The diameters of tumors varied from 1cm to 4 cm. Histologically, papillary and cystic architecture were present at least focally in all tumors. The papillae were covered by small to medium-sized cuboidal cells with abundant clear cytoplasm and often showed extensive secondary branching, which were often folded and densely packed, resulting in a solid appearance. The nuclei were round and uniform in shape; nucleoli were not prominent (Fuhrman grade 1 or 2). Neither mitotic figures nor necrosis was present. All 11 cases exhibited moderate to strong positivity for CK7, CA9, vimentin, and HIF-1α, coupled with negative reactions for CD10, P504S, and TFE3. Ksp-cadherin was positively expressed in 8 cases.VHL gene mutations were not found in all 11 cases. Losses of chromosomes 3 (monoploid chromosome 3) was detected in 3 cases.

CONCLUSIONSCCPRCC is uncommon and seemed to be an indolent tumor. The differential diagnosis should be included tumors, which harbor clear cell and papillary structure including clear cell renal cell carcinoma, papillary renal cell carcinoma, Xp11 translocation renal cell carcinoma, and CCPRCC. Immunohistochemical and molecular analysis may be help for its diagnosis.

Adult ; Aged ; Carcinoma, Renal Cell ; chemistry ; genetics ; pathology ; ultrastructure ; Chromosomes, Human, Pair 3 ; Female ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; analysis ; Kidney Neoplasms ; chemistry ; genetics ; pathology ; ultrastructure ; Male ; Middle Aged ; Mutation ; Prognosis ; Racemases and Epimerases ; analysis ; Translocation, Genetic ; Tumor Burden