Bronchi ; drug effects ; pathology ; Cell Line ; Cell Transformation, Neoplastic ; drug effects ; Epithelial Cells ; drug effects ; pathology ; Humans ; Mineral Fibers ; toxicity

Tricholoma giganteum is a famous species product of Jishou in the wes t of Hunan.It have not only special biological characteristics such as formatio n and ecological enviromment,but also delicious,fragrant with abundant nutrien t.It's mycelium can be isolated and growing well in medium,but its fruiting bo dy is difficult to be formed.

BACKGROUND: Oxygen-glucose depdvation (OGD) in cultured neurons simulates stroke to a certain degree and plays an important role in studying processing and pathophysiological mechanism of ischemic neuronal injury.OBJECTIVE: To produce experimental OGD models in cultured neurons.DESIGN, TIME AND SETTING: A grouping controlled study was performed at the Center Laboratory of Third Hospital, Peking University from January 2007 to March 2008.MATERIALS: Fetal Wistar rats with gestational age of 17-19 days were collected in this study.METHODS: Primary cultures of cortical neurons that were derived from fetal Wistar rats with gestational age of 17-19 days were performed to remove pollutional non-neuronal cells. OGD was produced by incubation with non-glucose balanced salt solution and 2% Oxyrasa in 7-day cultured cortical neuron cultures. Cell cultures were kept in a humidified 37 ℃ incubator. In the control group, cell culture medium was replaced with balanced salt solution containing 20 mmol/L glucose. In the sham operation group,balanced salt solution containing 20 mmol/L glucose and Oxyrasawere used to replace the medium.MAIN OUTCOME MEASURES: Oxygen concentration in the culture medium was measured with blood gas analysis; neuronal death in the experimental group was observed under phase contrast microscope; lactate dehydrogenase activity was detected with lactate dehydrogenasa assay; effect of oxygen-glucose deprivation on neuronal viability was observed with trypan blue staining.RESULTS: Measurement of oxygen concentration showed that hypoxia could be quickly achieved shortly after the addition of Oxyrase; lactate dehydrogenase assay revealed that after treatments of neuron cultures with Oxyrase and non-glucose balanced salt solution, lactate dehydrogenase release increased significantly with the treatment time; trypan blue staining and phase contrast microscope showed that cell viability decreased after treatments of Oxyrase and non-glucose balanced salt solution, and most neurons died 6 hours after OGD.CONCLUSION: These results show that Oxyrase, together with non-glucose balanced salt solution, can be conveniently used to produce OGD condition in cultured neuronal cells which is greatly useful in the study of simulating cerebral ischemia in vitro.

Aim To establish an UPLC-ESI-MS method for determination of asiaticoside and investigate its application to pharmacokinetic study in rats.Methods Eight rats were given 40 mg·kg~(-1) asiaticoside iv respectively.Drug plasma concentration was determined by UPLC-ESI-MS.Pharmacokinetic parameters were evaluated.Results Calibration curves were linear over 0.038～7.6 mg·L~(-1) and LLOQ was 38 μg·L~(-1),the recoveries of asiaticoside from plasma were larger than 95%,and RSD of inter-day and intra-day assay were below 10%.After iv administration of 40 mg·kg~(-1) asiaticoside,the pharmacokinetic parameters of AUC(0-t),T(1)/(2)β,CL,Vd were (81 443.67±57 156.81) μg·L~(-1)·min~(-1),(23.44±9.60) min,(0.19±0.07) L·min~(-1)·kg~(-1),(8.92±6.68) L·kg~(-1),respectively.Conclusion The method described in this report was sensitive and specific,and suitable for pharmacokinetic studies of asiaticoside in rats.

OBJECTIVETo investigate the expression of zinc finger E-box binding homebox 1 (ZEB1) in the prepuce of hypospadias children and its relationship to the incidence of hypospadias.

METHODSPrepuce tissues were collected from 37 children aged 6-15 months undergoing hypospadias repair and 11 age-matched controls receiving circumcision. Based on the position of the urethral meatus, the hypospadias cases were classified as severe (n = 13) and mild-moderate (n = 24). The mRNA and protein expressions of ZEB1 were determined by immunohistochemistry and RT-PCR.

RESULTSThe expression of the ZEB1 protein was remarkably higher in the severe (100% [13/13]) and mild-moderate hypospadias patients (75.0% [18/24]) than in the controls (9.1% [1/11]), with statistically significant differences between any two groups (P < 0.05). RT-PCR showed the integrated density value (IDV) of the ZEB1 mRNA expression to be (0.67 ± 0.21), (0.81 ± 0.24), and (1.55 ± 0.29) in the control, mild-moderate, and severe hypospadias patients, respectively, significantly higher in the severe hypospadias than in the control and mild-moderate hypospadias groups (P < 0.05), but with no significant difference between the latter two (P = 0.64).

CONCLUSIONThe expression of ZEB1 is significantly increased in hypospadias patients, and its upregulation is positively correlated with the severity of hypospadias, which suggests that the overexpression of ZEB1 may contribute to the development of hypospadias.

Biomarkers ; metabolism ; Case-Control Studies ; Circumcision, Male ; Foreskin ; metabolism ; Homeodomain Proteins ; genetics ; metabolism ; Humans ; Hypospadias ; classification ; etiology ; metabolism ; Immunohistochemistry ; Infant ; Male ; Penis ; RNA, Messenger ; metabolism ; Transcription Factors ; genetics ; metabolism ; Up-Regulation ; Urethra ; Zinc Finger E-box-Binding Homeobox 1

OBJECTIVETo study the relationship of the expression of FOXA1 in the prostate cancer (PCa) tissue with the Gleason score and clinical staging of PCa and with castration-resistant PCa (CRPC).

METHODSUsing the immunohistochemical method, we detected the expressions of FOXA1 and Ki-67 in the pathological sections of 35 cases of PCa and 21 cases of benign prostatic hyperplasia (BPH). Then we analyzed their correlation with the Gleason score and TNM staging of PCa and that with CRPC.

RESULTSThe positive expression of FOXA1 was significantly higher in the PCa than in the BPH tissue (P < 0.001) and was positively correlated with that of Ki-67 (P < 0.001) as well as with the Gleason score (P = 0.027) and clinical staging of PCa (P = 0.002), but showed no correlation with CRPC (P = 0.391).

CONCLUSIONThe positive expression of FOXA1 is increased in PCa, most significantly in the advanced stage of the tumor.

Disease Progression ; Hepatocyte Nuclear Factor 3-alpha ; metabolism ; Humans ; Ki-67 Antigen ; metabolism ; Male ; Neoplasm Grading ; Neoplasm Proteins ; metabolism ; Prostatic Hyperplasia ; metabolism ; pathology ; Prostatic Neoplasms ; metabolism ; pathology

Objective To evaluate the outcomes achieved by using left internal mammary artery(LIMA) to radial artery (RA) total arterial composite grafts in minimally invasive direct coronary artery bypass grafting (MIDCAB) for patients with multiple vessel disease.Methods From January 2009 to September 2015, 39 patients(24 males) with multiple vessel disease underwent MIDCAB with LIMA-RA total arterial composite grafts without cardiopulmonary bypass in our hospital .MIDCAB was performed through a left anterior minithoracotomy .Results All patients successfully underwent MIDCAB with LIMA-RA total arterial composite grafts.No patient required to convert to strenotomy during the surgery.Mean operation time was(176.1 ± 14.1)min.Revascularization was performed for 2 target vessels in 11 cases, 3 target vessels in 25 cases and 4 target vessels in 3 cases.Mean postoperative ventilation time was(21.9 ±27.9) h.Mean ICU time was(2.8 ±2.1) days, and mean postoper-ative inhosptial time was(11.2 ±3.3)days.There was no early death in perioperation.At a follow-up of 6 to 86 months[aver-age(27.5 ±18.0) months], one patient died.The overall survival at 2 years postoperatively was(96.0 ±3.9)%.The paten-cy rate of LIMA was 100%.The overall patency rate of RA grafts at 2 years postoperatively was(91.8 ±4.0)%.Conclusion MIDCAB with LIMA-RA total arterial composite grafts is a safe and effective procedure with favorable early and mid-term out-comes for patients with multiple vessel disease .

Objective To discuss function of the fusion cells of human bone marrow stromal cells (BMSCs) and human myeloma cell RPMI 8226 in the pathogenesis of multiple myeloma bone disease.Methods The cells,labeled by cell tracer green fluorescent probe (CMFDA) and red fluorescent probe (CMTMR),respectively,were induced into fusion by chemical polyethylene glycol (polyethyleneglycol,PEG-1000),and cell fusion model was set up.Whether fusion cells had nucleus fusion was determined by Karyotype analysis.The expressions of stemness-related genes,SIRP α gene and DC-STAMP gene in fusion cells were identified.Results Polyethylene glycol (PEG-1000) could mediate the integration of BMSCs and RPMI 8226 cells.The number of chromosomes in more than 80 ％ the hybrid cells was about 80.Fusion cells not only showed that BMSCs,stemness-related genes of c-myc,Klf-4 and OCT-4 genes expressed positively,but also the fusion-related genes SIRPα and DC-STAMP expressed positively.Conclusion BMSCs and RPMI 8226 cells can form fusion cells,and the cells have the potential for further integration,which is one of the important reasons for the promotion of muhiple myeloma bone destruction.

AIM: To investigate the neuroprotective effect of tubuloside B,one of the phenylethanoids isolated from the stems of Cistanche salsa,on H2O2-induced injury in PC12 cells.METHODS: PC12 cells were exposed to various doses of tubuloside B for 12 h,then treated with H2O2 at concentration of 100 ?mol/L for 24 h.The cell viability was observed with MTT assay.Reactive oxygen species and the mitochondrial membrane potential were measured with laser scanning confocal microscopy(LSCM).The DNA content and percentage of apoptosis were assayed by DNA agarose gel electrophoresis and flow cytometry.The activation of caspase-3 was detected with the caspase-3 activity assay kit.RESULTS: Following treatment with H2O2 for 24 h,H2O2 induced a significant decrease in cell viability;DNA ladder was observed and apoptosis percentage was as high as 48.0%.Accumulation of intracellular ROS,increase in caspase-3 activity and the decrease in mitochondrial membrane potential as indicated with the decrease of red/green ratios(from 5.97 to 0.41) were detected.However,pretreatment with tubuloside B(1,10 or 100 mg?L-1) for 12 h exhibited cytoprotective effects in a dose-dependent manner.Tubuloside B obviously enhanced the cell viability,reduced formation of the DNA ladder,and significantly reduced the number of cells labeled with Annexin-V.The percentage of apoptosis/necrosis neurons was significantly decreased to 30.9%,18.3% and 6.2%,respectively.LSCM showed that the tubuloside B attenuated the accumulation of ROS and the H2O2-induced collapse of mitochondrial membrane potential in PC12 cells.The significant decrease in caspase-3 activity was detected,compared to the H2O2-treated cells at the same time point.CONCLUSION: Tubuloside B has the neuroprotective capacity to antagonize H2O2-induced apoptosis and injury in PC12 cells,indicating it may be useful for treating some neurodegenerative diseases.

G (E165E) in exon 1 of KRT6A gene, were found in this patient. Conclusions A novel single nucleotide polymorphism of KRT16 gene which can result in the change of amino acid sequence is firstly reported and some known single nucleotide polymorphisms in KRT16 and KRT6A genes are also found in this study.