OBJECTIVETo explore the expression of pituitary tumor transforming gene (PTTG) in human renal clear cell carcinoma (RCCC) and its significance.

METHODSPTTG protein expression was detected by immunohistochemistry in 87 RCCC and 45 paired normal renal tissues.

RESULTSPTTG was expressed differentially between the normal renal and RCCC tissues. Compared with normal tissues, the primary RCCC tissues showed significantly increased expression of PTTG protein (P<0.001). The positive rate of PTTG protein was positively correlated to the tumor size, clinical stage and Fuhrman grade of RCCC (P=0.009, 0.008 and 0.035, respectively).

CONCLUSIONIncreased PTTG protein expression may be involved in the carcinogenesis and progression of RCCC.

Adult ; Aged ; Carcinoma, Renal Cell ; metabolism ; pathology ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Immunohistochemistry ; Kidney Neoplasms ; metabolism ; pathology ; Male ; Middle Aged ; Neoplasm Proteins ; genetics ; metabolism ; Securin

OBJECTIVETo study the effect of laminarin sulphate (LAMS) on the expressions of PTEN and P27kip1 in androgen-independent prostate cancer PC-3 cells in vitro, and investigate the mechanism of its anti-tumor action.

METHODSThe inhibitory effects of different concentrations of LAMS (0, 50, 100, 200 microg/ml) on androgen-independent prostate cancer PC-3 cells were detected by WST-8 assay. The morphology of PC-3 cells was observed under the fluorescence microscope, and the cell cycle and apoptosis were analyzed by flow cytometry. The mRNA and protein levels of PTEN and P27kip1 were measured by RT-PCR and Western blot.

RESULTSLAMS inhibited the proliferation of androgen-independent prostate cancer PC-3 cells in a dose- and time-dependent manner, and the cell cycle analysis showed that PC-3 cells were arrested in the S phase after treated with different concentrations of LAMS. The rate of apoptosis was increased and many typical apoptotic morphological features were observed under the fluorescence microscope. The PTEN and P27kip1 expressions at mRNA and protein levels were increased in a dose-dependent manner.

CONCLUSIONLAMS can inhibit the proliferation, arrest the cell cycle in the S phase and induce apoptosis of prostate cancer PC-3 cells. The significantly increased expressions of PTEN and P27kip1 may be one of the mechanisms for LAMS inhibiting prostate cancer PC-3 cells.

Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Line, Tumor ; drug effects ; Cell Proliferation ; drug effects ; Cyclin-Dependent Kinase Inhibitor p27 ; genetics ; Humans ; Male ; PTEN Phosphohydrolase ; genetics ; Polysaccharides ; pharmacology ; Prostatic Neoplasms ; blood ; genetics ; RNA, Messenger ; genetics

OBJECTIVETo investigate the effects of adenovirus-mediated PTEN and P27 on the invasion of PC-3 in vitro and angiogenesis, along with their synergy in the treatment of prostate cancer.

METHODSRecombinant adenovirus vectors of the human tumor suppressor genes PTEN and P27 were constructed. The replication-incompetent recombinant adenovirus was packaged and propagated in HEK293 cells. The viral titer was examined by plaque assay and the mRNA and protein expressions of PTEN and P27 in human prostate cancer cell line PC-3 infected with Ad-PTEN and Ad-P27 were determined by RT-PCR and Western blot respectively. The invasion of PC-3 cells in vitro was examined by Boyden chamber assay. MTT assay was used to testify the effect of supernatant from PC-3 infected with Ad-PTEN and Ad-P27 on the proliferation of endothelial cells ECV-304 and the CAM test was used to testify the effect of PTEN and P27 on angiogenesis. The difference between the combined therapy group and the single gene therapy group was also examined.

RESULTSThe viral titers of Ad-PTEN and Ad-P27 were 1.8 x 10(7) pfu/ml and 1.2 x 10(9) pfu/ml respectively. Adenovirus infection verified that the mRNA and protein expression of PTEN and P27 were steady in human PC-3 cells. The invasion in vitro of PC-3 cells was significantly inhibited by infection with Ad-PTEN or/and Ad-P27. CAM and MTT assays of ECV-304 confirmed that the supernatant from PC-3 cells infected with Ad-PTEN or/and Ad-P27 could inhibit the angiogenesis effectively. There was a significant difference between the combined therapy group and the single gene therapy group.

CONCLUSIONThe combined gene therapy of Ad-PTEN and Ad-P27 plays a synergistic role in inhibiting the invasiveness of PC-3 cells and angiogenesis.

Adenoviridae ; genetics ; Cell Line, Tumor ; Cyclin-Dependent Kinase Inhibitor p27 ; genetics ; Humans ; Male ; Neoplasm Invasiveness ; PTEN Phosphohydrolase ; genetics ; Prostatic Neoplasms ; blood supply ; pathology ; Transfection

OBJECTIVETo study the effects of MTA1 small interfering RNA (siRNA) on the anchorage-independent growth and anoikis of prostate cancer cell line PC-3.

METHODSAfter transfection of human prostate cancer PC-3 cells by MTA1 siRNA, we detected the expression of the MTA1 gene by real-time PCR and Western blot, the anchorage-independent growth of the cells by clone formation in soft agar, and their anoikis by DNA fragmentation assay and flow cytometry.

RESULTSCompared with the control group, MTA1 siRNA transfection significantly decreased the mRNA and protein levels of MTA1, inhibited the anchorage-independent growth of the PC-3 cells, and induced their anoikis, all in a dose- and time-dependent manner (r = 0.935, P = 0.001; r = 0.901, P = 0.0005; r = 0.916, P = 0.0003).

CONCLUSIONMTA1 siRNA can inhibit the anchorage-independent growth of prostate cancer cells by inducing their anoikis.

Anoikis ; genetics ; Gene Expression Regulation, Neoplastic ; Histone Deacetylases ; genetics ; Humans ; Male ; Prostatic Neoplasms ; genetics ; RNA, Small Interfering ; Repressor Proteins ; genetics ; Transfection ; Tumor Cells, Cultured

Objective:To explore the value of echocardiography for diagnosing infectious endocarditis (IE).Methods:A total of 487 patients with cardiovascular implantable electronic devices (CIED) infection treated in our hospital from 2013-01 to 2015-06 were enrolled.Based on symptoms,blood culture and echocardiography,9 patients with suspected IE were further examined by 18F-FDG PET-CT to confirm their diagnosis and classification.Definitive therapy was conducted and the patients were followed-up for 1 year to confirm the diagnostic accuracy of echocardiography on CIED induced IE.Results:3 patients were preliminarily diagnosed for bacteremia since no vegetation was found by echocardiography,while IE was finally diagnosed by PET-CT.2 patients were preliminarily diagnosed for IE by echocardiography presented valvular vegetation,while PET-CT showed no evidence of vegetation;then one of them was diagnosed as bacteremia by positive blood culture and another was diagnosed as non-infection.4 patients were preliminarily diagnosed for IE by echocardiography indicated existing vegetation after CIED lead extraction,while PET-CT demonstrated no infection sign in heart chamber and the finally diagnosed was as "non-infectious fibrous residual tissue".According to final diagnosis,definitive therapies were performed to specific patients with at least 1 year follow-up study,no one had new and recurrent infection.Conclusion:Echocardiography had deficiency for diagnosing vegetation in heart chamber especially in suspicious IE patients after CIED lead extraction.It is necessary to make accurate diagnosis with other method for guiding appropriate therapy.

Myasthenia gravis (MG) is a prototypical antibody-mediated neurological autoimmune disease with the involvement of humoral immune responses in its pathogenesis. T follicular helper (Tfh) cells have been implicated in many autoimmune diseases. However, whether and how Tfh cells are involved in MG remain unclear. Here, we established and studied a widely-used and approved animal model of human MG, the rat model with acetylcholine receptor alpha (AChRα) subunit (R-AChR)-induced experimental autoimmune myasthenia gravis (EAMG). This model presented mild body-weight loss 10 days after the first immunization (representing the early stage of disease) and more obvious clinical manifestations and body-weight loss 7 days after the second immunization (representing the late stage of disease). AChR-specific pre-Tfh cells and mature Tfh cells were detected in these two stages, respectively. In co-cultures of Tfh cells and B cells, the number of IgG2b-secreting B cells and the level of anti-AChR antibodies in the supernatant were higher in the cultures containing EAMG-derived Tfh cells. In immunohistochemistry and immunofluorescence assays, a substantial number of CD4/Bcl-6 T cells and a greater number of larger germinal centers were observed in lymph node tissues resected from EAMG rats. Based on these results, we hypothesize that an AChR-specific Tfh cell-mediated humoral immune response contributes to the development of EAMG.
Animals ; B-Lymphocytes ; immunology ; Disease Models, Animal ; Female ; Immunity, Humoral ; Lymph Nodes ; immunology ; Myasthenia Gravis, Autoimmune, Experimental ; immunology ; Protein Subunits ; immunology ; Proto-Oncogene Proteins c-bcl-6 ; immunology ; Rats, Inbred Lew ; Receptor Cross-Talk ; Receptors, Cholinergic ; immunology ; T-Lymphocytes, Helper-Inducer ; immunology

OBJECTIVE: To obtain the subtypes of the clinical hypertension population based on symptoms and to explore the relationship between hypertension and comorbidities. METHODS: The data set was collected from the Chinese medicine (CM) electronic medical records of 33,458 hypertension inpatients in the Affiliated Hospital of Shandong University of Traditional Chinese Medicine between July 2014 and May 2017. Then, a hypertension disease comorbidity network (HDCN) was built to investigate the complicated associations between hypertension and their comorbidities. Moreover, a hypertension patient similarity network (HPSN) was constructed with patients' shared symptoms, and 7 main hypertension patient subgroups were identified from HPSN with a community detection method to exhibit the characteristics of clinical phenotypes and molecular mechanisms. In addition, the significant symptoms, diseases, CM syndromes and pathways of each main patient subgroup were obtained by enrichment analysis. RESULTS: The significant symptoms and diseases of these patient subgroups were associated with different damaged target organs of hypertension. Additionally, the specific phenotypic features (symptoms, diseases, and CM syndromes) were consistent with specific molecular features (pathways) in the same patient subgroup. CONCLUSION The utility and comprehensiveness of disease classification based on community detection of patient networks using shared CM symptom phenotypes showed the importance of hypertension patient subgroups.

OBJECTIVE: Larotaxel is a new chemical structure drug, which has not been marketed worldwide. Accordingly, the standard identification and quantification methods for larotaxel remain unclear. The spectrometric analyses were performed for verifying weight molecular formula, molecular weight and chemical structure of larotaxel. Besides, a quantification method was developed for measuring larotaxel in the liposomes. METHODS: The molecular formula, molecular weight and chemical structure of larotaxel were studied by using mass spectrometry (MS), infra-red (IR), nuclear magnetic resonance (NMR) and ultraviolet-visible (UV-vis) spectrometric techniques. The absorption wavelength of larotaxel was investigated by UV-vis spectrophotometry full-wavelength scanning. Besides, a quantification method was developed by high performance liquid chromatography (HPLC), and then validated by measuring the encapsulation efficacy of larotaxel liposomes. RESULTS: The four spectral characteristics of larotaxel were revealed and the corresponding standard spectra were defined. It was confirmed that larotaxel had the structure of tricyclic diterpenoids, with the molecular formula of C45H53NO14, the molecular weight of 831.900 1, and the maximum absorption wavelength of 230 nm. The quantitative method of larotaxel was established by using HPLC with a reversed phase C18 column (5 μm, 250 mm×4.6 mm), a mobile phase of acetonitrile-water (75:25, volume/volume), and a detection wavelength of 230 nm. The validation study exhibited that the established HPLC method was stable, and had a high recovery and precision in the quantitative measurement of larotaxel in liposomes. In addition, a new kind of larotaxel liposomes was also successfully prepared. The particle size of the liposomes was about 105 nm, with an even size distribution. And the encapsulation efficiency of larotaxel in the liposomes was above 80%. CONCLUSION The present study offers reference standard spectra of larotaxel, including MS, IR, NMR, and UV-vis, and confirms the molecular formula, molecular weight and chemical structure of larotaxel. Besides, the study develops a rapid HPLC method for quality control of larotaxel liposomes.
Chromatography, High Pressure Liquid ; Liposomes ; Magnetic Resonance Spectroscopy ; Taxoids