Objective To localize cytochrome P450 enzymes 4A and 2C in central nervous cells of normal male rats.Methods Eight drug/alcohol untreated normal male rats (150-200 g of body weight) were treated by the optimized perfusion technique, then brain tissues were postfixed, paraffin-embedded and cut into series sections, which were labeled by the improved strept-avidin-biotin complex DAB-nickel enhancer (SABC-DAB-Ni) immunohistochemistry and hematoxylin & eosin (H & E) stain techniques.Results The immunohistochemical results indicated that P450 2C-11 enzyme was localized in diverse numbers of neurons as well as some neuroglial cells, with focal or defuse distribution in many brain regions such as cerebrum, thalamus, olfactory bulb, hypothalamus, brain-stem, hippocampus, cerebellum, interpositus nucleus, caudate-putamen, and globus pallidus. In contrast, no positive findings of P450 4A-2, 3 and 8 enzymes were obtained in the same animals. With high magnification, 2C-11 protein was able to be roughly observed on the endoplasmic reticulum of the rat neurons.Conclusions P450 2C-11 protein, rather than P450 4A-2, 3 and 8, may be a candidate of brain P450 enzymes in the normal male rats.

BACKGROUND: The blood oxygen saturation is an important physiological parameter of human hfe; it can be affected by multiple factors due to the imbalance of oxygen supply and consumption, and consequently, the normal physical metabolism is affected. The existing manufacturing techniques are complex and are used with higher demands. OBJECTIVE: To investigate the accuracy of the novel reflecting oximeter designed on the basis of reflecting principle in monitoring the variance of blood oxygen saturation with the change of its concentration. DESIGN: To introduce the structure, working principle and design of the reflecting oximeter and make comparative study with adult rabbits as the subjects. SETTING: Cardiopolmonary Bypass Department of Cardiovascular disease hospital of the Chinese Academy of MedicalSciences. MATERIALS: The study was performed at the Departments of Anesthesiology and Cardiopulmonary Bypass of the Cardiovascular Disease Hospital,Chinese Academy of Medical Sciences from August to October in 2001.Ten adult rabbits were selected. METHODS: The oximeter consists of a converter, a driver, a central processing mould and liquid-crystal monitor. It is characterized by the onethree optical cable connected with reflecting probe, postcentral management system and digital displayer. This oxmeter was developed basing on the basis of fixed relationship between blood oxygen saturation and radiant absorbency (or reflecting). In this study,cardiovascular-pulmonary bypass was established on rabbits that were then exposed to oxygen and NO mixture supply of different concentrations,and then the corresponding blood oxygen saturations were determined with the new oximeter and compared with series-wound Baite saturation meter and blood-gas analysis. MAIN OUTCOME MEASURES: The blood oxygen saturation at different concentrations determined with the reflecting oximeter and Baite saturation meter. RESULTS: At the same oxygen concentration, the blood oxygen saturation determined with the new measure was not significantly different from that determined with Baite blood oxygen saturation meter(P＞0.05 ). CONCLUSION: The variance of blood oxygen saturation with the change of oxygen concentration can be detected by the reflecting oximeter with the result similar to that by Baite blood oxygen saturation meter.

The growing status of Anoectochilus roxburghii seedling was observed and the survival rate of seedlings, height, stem diameter and plant fresh weight under the conditions of different transplanting substrate compositions, planting density, shading rate were measured. The results showed that the effects of different transplanting substrates, planting densities, shading rates and nutrient solutions on the growing status of A. roxburghii plantlets varied greatly. A. roxburghii plantlets demonstrated a high survival rate and better growing status under the Following conditions: the ratio of peat and river sand as 2: 1, the planting density as 3 cm x 3 cm, the shading rate as 70%, and the nutrient solution as 1/4MS. The findings of the study provide a solid technical solution for the artificial cultivation of A. roxburghii plantlets.
Breeding ; methods ; Culture Media ; chemistry ; pharmacology ; Orchidaceae ; drug effects ; growth & development ; Seedlings ; drug effects ; growth & development ; Survival Analysis

Background:Anomalous origin of the left coronary artery from the pulmonary artery (ALCAPA) is a rare but potentially life?threatening congenital heart defect. A retrospective analysis was carried out to elucidate the surgical outcomes of ALCAPA in infants and children using follow?up echocardiography. Methods: From September 2008 to March 2017, 26 children diagnosed with ALCAPA underwent left coronary re?implantation. All surviving patients received echocardiography during follow?up. Results: The mortality rate after the operation was 11.5%. Before repair, twenty patients (76.9%) presented with left ventricular (LV) dysfunction. The mean Z?score of the preoperative LV end?diastolic diameter was 4.42 ± 2.09. Mitral regurgitation (MR) was present in all patients. Two patients (7.7%), both with mitral valve prolapse, underwent mitral valve repair at the time of ALCAPA repair. Two children required postoperative extracorporeal membrane oxygenation. LV function normalized at a median time of 5.3 months (range:0.5–36.0 months). The Z?score of the LV end?diastolic diameter decreased simultaneously. The degree of MR gradually decreased in all surviving patients.All patients had patency of the proximal left coronary artery confirmed by echocardiography at the most recent follow?up. Six patients (26.1%) showed supravalvar pulmonary stenosis and seven patients (30.4%) showed right pulmonary stenosis during follow?up. Conclusions: Coronary re?implantation was effective for rebuilding a dual coronary system in patients with ALCAPA and resulted in progressive improved LV function and reduced functional MR. Echocardiography was valuable for evaluating the outcomes. LV function, the degree of MR, and possible complications could be detected with follow?up echocardiography.

OBJECTIVETo establish a nasopharyngeal carcinoma (NPC) cell line CNE1-pLVTHM/BART7 with stable ebv-miR-BART7 overexpression.

METHODSThe recombinant lentivirus pLVTHM/BART7 expression plasmid was packaged into mature lentivirus by 293FT cells and used to infect CNE1 cells. Flow cytometry was employed for sorting the GFP(+) cells. The efficiency of ebv-miR-BART7 overexpression was determined using qRT-PCR.

RESULTSThe recombinant lentivirus plasmid pLVTHM/BART7 was successfully constructed and verified by PCR and sequencing. The expression of ebv-miR-BART7 in CNE1 cells infected with the lentivirus pLVTHM/BART7 was significantly increased as compared with the negative control and the blank control cells.

CONCLUSIONThe recombinant lentivirus vector pLVTHM/BART7 results in high and stable expression of ebv-miR-BART7 in infected CNE1 cells, which provides a useful cell model for further studies of the role of ebv-miR-BART7 in nasopharyngeal carcinoma.

Carcinoma ; Cell Line, Tumor ; Genetic Vectors ; Humans ; Lentivirus ; genetics ; MicroRNAs ; Nasopharyngeal Neoplasms ; genetics ; Plasmids

OBJECTIVETo investigate the intracellular trafficking pathway of fusion protein epidermal growth factor-adenovirus early region 4 open reading frame 4 protein (EGF-E4orf4) internalized via epidermal growth factor (EGF) receptor and its affectivity on extracellular signal-regulated kinase (ERK) phosphorylation.

METHODSMDA-MB-231 and BGC823 cells were incubated with fluorescein isothiocyanate-EGF-E4orf4 or EGF at different time points. The specific molecular mark of early endosome or late lysosome was labeled by indirect immunofluorescence, and then colocalization staining was observed using confocal laser microscopy. The levels of ERK phosphorylation were detected by Western blot.

RESULTSThe fluorescent signal of fusion protein EGF-E4orf4 accumulated within the cells and congregated to the perinuclear region. A nucleus localization of the fusion protein was only at MDA-MB-231 cell. Colocalization of EGF-E4orf4 with early endosome/late lysosome was observed. EGF-E4orf4 stimulated ERK phosphorylation, which was most obvious 10 minutes after stimulation, and then gradually attenuated, which was similar to EGF stimulation but with a less decrease.

CONCLUSIONSInternalized EGF-E4orf4 can be slowly degraded via endosome-lysosome pathway. The action features of EGF-E4orf4 are remarkably different between MDA-MB-231 and BGC823 cells, which may help explain the differences in its anti-tumor potency and in the special selectivity toward different tumor cells.

Adenovirus E4 Proteins ; pharmacokinetics ; Cell Line, Tumor ; Epidermal Growth Factor ; pharmacokinetics ; Extracellular Signal-Regulated MAP Kinases ; metabolism ; Humans ; Phosphorylation ; drug effects ; Protein Transport ; Recombinant Fusion Proteins ; pharmacokinetics ; Viral Proteins ; pharmacokinetics

BACKGROUNDAnomalous origin of the left coronary artery from the pulmonary artery (ALCAPA) is a rare but potentially life-threatening congenital heart defect. A retrospective analysis was carried out to elucidate the surgical outcomes of ALCAPA in infants and children using follow-up echocardiography.

METHODSFrom September 2008 to March 2017, 26 children diagnosed with ALCAPA underwent left coronary re-implantation. All surviving patients received echocardiography during follow-up.

RESULTSThe mortality rate after the operation was 11.5%. Before repair, twenty patients (76.9%) presented with left ventricular (LV) dysfunction. The mean Z-score of the preoperative LV end-diastolic diameter was 4.42 ± 2.09. Mitral regurgitation (MR) was present in all patients. Two patients (7.7%), both with mitral valve prolapse, underwent mitral valve repair at the time of ALCAPA repair. Two children required postoperative extracorporeal membrane oxygenation. LV function normalized at a median time of 5.3 months (range: 0.5-36.0 months). The Z-score of the LV end-diastolic diameter decreased simultaneously. The degree of MR gradually decreased in all surviving patients. All patients had patency of the proximal left coronary artery confirmed by echocardiography at the most recent follow-up. Six patients (26.1%) showed supravalvar pulmonary stenosis and seven patients (30.4%) showed right pulmonary stenosis during follow-up.

CONCLUSIONSCoronary re-implantation was effective for rebuilding a dual coronary system in patients with ALCAPA and resulted in progressive improved LV function and reduced functional MR. Echocardiography was valuable for evaluating the outcomes. LV function, the degree of MR, and possible complications could be detected with follow-up echocardiography.

Objective To investigate the clinical features of unexpected sudden death (SUD) clustered in families in Yunnan province.Methods This retrospective study analyzed the clinical features of SUD occurred between July to September 2005 in 7 families in Yunnan province.Results All 16 SUD patients shared common clinical features such as fatigue and repeated syncope and one group of SUD patients (n = 8 from 4 families) presented with the gastric intestinal tract manifestations including nausea, vomiting, abdominal pain and diarrhea with suspected dietary history and abnormal laboratory enzyme findings (GOT/ GPT,CK/CKMB,LDH/LDH1 etc.).In SUD patients without gastric intestinal tract manifestations (n = 8 from 3 families), there were no clear symptoms before death and repeated ventricular tachycardia and ventricular fibrillation were recorded in one survivor.There was no clear evidence for the involvements of hereditary and infectious factors for observed SUD.conclusion The reason for the unexpected sudden death clustered in 7 families in Yunnan remains unclear.Repeated syncope and fatigue served as the common clinical features in the presence or absence of gastric intestinal tract manifestations in all SUD cases.Further studies are needed to clarify the pathology and detailed clinical manifestations of SUD occurred in this area.