AIM: To observe the effect of protein kinase C-?(PKC?)antisense oligonucleotide on cell growth, cell cycle and the expression of cyclin E in human poor-differentiated nasopharyngeal carcinoma(NPC) cell line CNE-2Z. METHODS: Antisense PKC? was transfected by cationic liposome(LP) in CNE-2Z cells to analyze the cell growth and cell cycle by MTT colorimetric assay and flow cytometry, respectively. Moreover, the expression of cyclin E was determined by immunocellularchemistry and scanning the result of dot-blotting. RESULTS: ①With the concentration of antisense PKC? increasing, the relative cell growth index was decreased gradually( P

Objective To investigate the effects of high-affinity tyrosine kinase receptors TrkB,TrkC and the low-affinity neurotrophin receptor p75 in spiral ganglion cell(SGC)of cisplatin-induced ototoxicity.Methods The 50 adult Wistar rats were divided randomly into 5 groups received intraperitoneal injection of cisplatin with vary dose.Control group was received equivalent volumes of saline.The group received l day intraperitoneal injection was cisplatin treated at a dose of 5 mg/kg and killed at next day.The group received 3 days was cisplatin treated for 3 days at sanle dose daily and then killed at next day.The group A received 5 days was cisplatin treated for 5 days and killed at next day.The group B received 5 days was cisplatin treated for 5 days and then were sacrificed after 7 davs.The change of mRNA level of neurotrophin receptors in cochlear tissue were examined bv RT-PCR.The expressing pattern of TrkB,TrkC,P75 in damaged cochlea were study by immunochemistry using antibodies against TrkB,TrkC,P75 protein.Results The research data showed the expression of Trk B,Trk C,D75 exhibited in SGC wag dynamic along with the administration lasting.The mRNA and protein level of Trk B(x±s)at day 1 and 3 after cisplatin treatment were 0.76±0.06,88.78±4.28,0.82±0.09 and 91.64±4.06,with significant difference among those and other groups(P＜0.05).The mRNA and protein level of TrkC at day 1 after cisplatin treatment were 0.80±0.05 and 89.55±2.76.with significant difference among that and other groups(P＜0.05).The mRNA and protein level of p75 at the control group and cisplatin treated groups were 0.64±0.04,55.16±3.10.0.77±0.04,78.46±3.86,1.01±0.09,105.02±6.61,1.18±0.09,111.10±6.08.0.51±0.04 and 42.74±5.20.with significant difference among the control group and cisplatin treated groups(P＜0.05).Conclusions The expression of Trk B increased to peak at day 1-3 after cisplatin treatment and decreased at day 5 early and following weeks.The expression of Trk C went up to peak at day l after cisplatin treatment and went down during subsequently time.P75 kept a trend of continuance increased during the drug treatment and decrease at drug stopped.The expression of Trk B.Trk C and P75 may be involved in cochlear insuh with cisplatin-induced.Trk B and Trk C may play an important role in the reparative process of cochlear,especially at early stage of the damage.P75 could promote SGC apoptosis in cisplatin-induced neuretoxicity.

AIM: To test the ability of multifocal visual evoked potential (mfVEP) in the detecting of glaucoma by comparing the mfVEP recorded from normal subjects and glaucoma patients.METHODS: The mfVEP of 32 normal eyes (n =21) and of 58 eyes (n =37) with primary glaucoma were recorded with the Vision Monitor electrophysical apparatus by the second kernel analysis and to determine the correlation of the topographic location between them.RESULTS: There were significant variability (the coefficient of variation was 43.05%) in mfVEP RMS amplitude in the normal subjects; The RMS amplitude of eyes with glaucoma were smaller than that of the normal eyes and significantly statistical difference were found in the relatively center (namely the 0° -10° ring zone) and in superior nasal quadrant (P＜0.05) while there were no significantly statistical differences of the latency time between them.CONCLUSION: The normal subjects have large individual variability of mfVEP responses. The RMS amplitude of the mfVEP of glaucomatous eyes descends, especially in center zone and superior nasal quadrant.

Objective To investigate the role of heterogeneous expression of calcium handling proteins and spatial heterogeneity of APD restitution in the maintenance mechanism of ventricular fibrillation.Methods During programmed electrical simulation, APD restitution curves in the endocardium and epicardium of LV were constructed by plotting APD100 and diastolic interval.APD ahernans,delayed afterdeporlarization events were recorded simultaneously.Endocardial and epicardial myocytes were isolated from LV base and apex.Real time-PCR and Western blotting were performed to determine the relative messenger RNA and protein expression levels of calcium handling proteins.Results The normal hearts have spatial heterogeneity of action potential restitution property and transmural heterogeneity of calcium handling proteins.The slopes of the APD restitution curve in the endocardium were significantly steeper than those in epicardium, and the slopes of APD curve at the LV apex were significantly steeper than those in LV base.However, delayed afterdeporlarization events with larger amplitude and earlier onset consistently occurred in the endocardium of LV base.After programmed electrical simulation,the expression of messenger RNA of RyR2,SERcA2a except for Calstabin2 significantly decreased(by 57%and 41%,respectively,P＜0.05)in the endocardium of the base, while the expression of RyR2, SERCA2a,Calstabin2 significantly increased (by 90%.78%and 64%,respectively,P＞0.05)in the epicardium of LV base.Although transmural heterogeneity of calcium handling proteins at the LV apex were also observed after rapid pacing,there is no significant differences in the transmural heterogeneity at the LV apex compared to the LV base.The base of LV has unique calcium handling properties.Conclusions It has been shown that Calcium cycling could modulate APD restitution property in the intact heart.The interaction between action potential and calcium dvnamics instabilities is one of the most important reasons why simple criterion such as the APD restitution slope ＞1 may fail to accurately predict the onset of APD alternans.

AIMTo detect the deletion distribution of dystrophin gene and dystrophin changes in muscle cells of the patients with Duchenne/Becker muscular dystrophy (DMD/BMD), furthermore to investigate the relationship between them and clinical symptoms.

METHODS42 patients with DMD/BMD were screened by 9 primers multiplex PCR. The patients from 5 DMD and 2 BMD were detected by immunofluorescence technique for analyzing dystrophin located in muscle cell membrane, compared with 2 normal males.

RESULTSThe deletion of one or more exons was found in 21 patients. 16 cases (76.2%) were detected in the central region and 5 patients (23.8%) in the 5' extreme region, especially in exon 48 (6 patients). Negative result of staining was seen in 5 DMD patients. Of these, one case of DMD had no detectable levels of dystrophin, but no deletion of DMD gene. Dystrophin immunostaining from two BMD patients consisted of a discontinuous staining pattern around most fibers.

CONCLUSIONIt might be possible that some correlation existed between the type of gene deletion and the degree of severity of the disease. The amount and size of exon deletion may not affect the symptoms. DMD/BMD are highly heterogeneous in clinical manifestation and in inheritance pattern. The pathologic foundation of DMD and BMD is the absence or abnormal expression of dystrophin. The consequence of that depends not only on the degree, but also on the function.

Adolescent ; Adult ; Child ; Child, Preschool ; Dystrophin ; analysis ; Exons ; Gene Deletion ; Humans ; Male ; Multiplex Polymerase Chain Reaction ; Muscular Dystrophy, Duchenne ; genetics ; Sequence Deletion ; Young Adult

OBJECTIVETo study transcription, expression, and bioactivity of expressive protein of vascular endothelial growth factor 165 (VEGF(165)) gene, after bone marrow stromal cells (BMSCs) were transferred by VEGF(165) gene mediated by adenovirus vector ex vivo.

METHODSBMSCs were Isolated and cultured by gradient centrifugation method. The cells cultured are transferred by recombinant adenovirus vector that carry VEGF(165) gene. Transcription and expression of VEGF(165) gene in BMSCs and secretion of VEGF protein in culture medium were measured by reverse transcriptase-polymerase chain reaction (RT-PCR), Western blot and enzyme linked immunosorbent assay (ELISA) methods. The activity of VEGF protein in culture media was detected by proliferation effects on vascular endothelial cells.

RESULTSWhen Ad. VEGF(165) transferred into BMSCs, there was an effective transcription and expression. The expressed product in the media of transferred cells had highly biological activity on proliferation of rat aortic vascular endothelial cells (P < 0.01).

CONCLUSIONSAdenovirus vector can be transferred into BMSCs efficiently and safely. Adenovirus mediated VEGF(165) gene transferred into BMSCs could express VEGF protein with highly biological activity, which provided foundation on BMSCs based gene therapy for ischemic disease.

Adenoviridae ; genetics ; Animals ; Bone Marrow Cells ; metabolism ; Cell Proliferation ; drug effects ; Cells, Cultured ; Endothelial Cells ; cytology ; drug effects ; Genetic Vectors ; Rats ; Rats, Sprague-Dawley ; Recombinant Proteins ; biosynthesis ; pharmacology ; Stromal Cells ; metabolism ; Transfection ; Vascular Endothelial Growth Factor A ; biosynthesis ; genetics ; pharmacology

Objective:To explore the prognosis and risk factors for conversion from off-pump coronary artery bypass grafting (OPCABG) to coronary bypass grafting (CABG) during surgery.
Methods: We retrospectively analyzed 2613 patients with elective OPCAB in our hospital from 2001 to 2012, there were 62 (2.37%) patients converted to CABG during the operation as Conversion group, the rest 2551 patients were set as Non-conversion group. The peril-operative baseline clinical data and prognosis condition were compared between 2 groups. The risk factors causing the in-operative conversion were studied with binary logistic regression analysis.
Results: The total conversion rate was 2.37%, including 42 patients of hemodynamic instability, 6 with dififculty of target vessel exposure, 9 with malignant arrhythmia, 3 with graft occlusion and 2 patients with other reasons. Compared with Non-conversion group, the Conversion group had increased post-operative drainage and ventilation time, higher rates of second thoracotomy for stop bleeding and higher peril-operative mortality. Binary logistic regression analysis indicated that chronic obstructive pulmonary disease, previous history of CABG, NYHA class≥3, LVEF≤40%and left main disease were the independent risk factors for in-operative conversion.
Conclusion: Conversion from OPCAB to CABG during the operation would be result in signiifcantly higher morbidity and mortality in relevant patients.

OBJECTIVETo investigate the clinical value of CT perfusion in diagnosing and assessing intracranial neoplasms and tumor-like lesions.

METHODS16-slice helical CT perfusion imaging was performed in 56 patients who were clinically suspected to have intracranial neoplasm or tumor-like lesion. With a GE-Light Speed 16-slice helical CT scanner, routine plain-CT scanning was performed to localize the central slice of the lesion. Perfusion imaging was then carried out using cine scan technique to maintain a slice thickness of 5-10 mm, a total dose of 50-70 ml of contrast-medium at an injection flow rate of 3-5 ml/s, a delay time of 7 s and a total scan time of 50 s. The images were processed using perfusion software in an ADW 4.0 workstation, meanwhile, time-density curves (TDC) of different kinds of lesions were also produced and analyzed.

RESULTSThe pathological types in this series included: 29 gliomas (12 low-grade and 17 high-grade), 2 ependemomas, 2 hemangioblastomas, 1 medulloblastoma, 2 metastatic tumors, 1 lymphoma, 5 meningiomas, 2 schwannomas, 1 germinoma, 1 teratoma in the pineal region, 6 cavernous hemangiomas, 2 inflammatory granulomas, 1 tuberculoma, and 1 hyperplasia of the choroid plexus. TDC of high-grade glioma, low-grade glioma and meningioma was different from each other. The cerebral blood flow (CBF), cerebral blood volume (CBV), particularly, the permeability surface (PS) value of glioma was found to increase significantly with the escalation of tumor differentiation grade. In PS map, margin of the tumor could be clearly showed, which was very useful when hemorrhaging within the tumor occurred. CBF in meningioma was lower than that in high-grade glioma, but there was no statistical difference in CBV, MTT and PS between these two types of tumor. The features of intracranial cavernous hemangioma such as significant prolongation of MTT, different TDCs, and zero perfused areas were diverse on CTP image, which was helpful in differentiating it from the other lesions. The germinoma and teratoma had rather low CBF and CBV value, but a remarkably high PS value, furthermore, they showed a rapid escalated TDC with a slowly and continuously elevated platform. The perfusion features of schwannoma was concordant with its pathological findings. However, no visible specific feature of inflammatory lesion was found on CTP image in this series.

CONCLUSIONMulti-slice helical CT perfusion imaging may be helpful in revealing histopathological features and hemodynamic changes as well as differential diagnosis of intracranial neoplasms and tumor-like lesions. When combined with other image and clinical information, CTP can play an important role in pre-operative diagnosis and treatment planning for intracranial neoplasms and tumor-like lesions.

Brain ; blood supply ; Brain Neoplasms ; diagnosis ; diagnostic imaging ; Cerebrovascular Circulation ; Diagnosis, Differential ; Glioma ; diagnosis ; diagnostic imaging ; Hemangioma, Cavernous ; diagnosis ; diagnostic imaging ; Humans ; Meningeal Neoplasms ; diagnosis ; diagnostic imaging ; Meningioma ; diagnosis ; diagnostic imaging ; Reproducibility of Results ; Sensitivity and Specificity ; Tomography, Spiral Computed ; methods

Adolescent ; Adult ; Female ; Follow-Up Studies ; Humans ; Male ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; drug therapy ; Treatment Outcome ; Young Adult

BACKGROUNDTo evaluate the influence ultrasonic processing on the aggregation of PrP(Sc) in the brain extracts prepared from the scrapie-infected hamsters, and to seek for the way to prepare lower molecular PrP(Sc) polymer.

METHODSThe extracts of infected brains were prepared with a lysis solution, and treated with ultrasonics at various conditions during the different phases. The distribution and aggregation state of PrP(Sc) were analyzed by proteinase K treated. Western blot, and afterwards, quantitatively calculated with a commercially supplied software Image Totaltech.

RESULTSThe amount of PrP(Sc) in the supernatant of brain homogenates was 1.29-to 1.58-fold increased with appropriate sonication (15 s for 30 times). At the same conditions of ultrasound, the PrP amount in the supernatant prepared from the scrapie-infected hamster brain was significantly increased, whereas that prepared from healthy animal used as normal control showed little change. Comparative analyses of PrP(Sc) pellets prepared by high-speed centrifugation revealed that about 90% PrP(Sc) released into supernatant after ultrasound processing.

CONCLUSIONAppropriate sonication of homogenate of scrapie-infected brain increases the extracted amount of PrP(Sc), being favorable to laboratory diagnosis. Larger molecular PrP(Sc) aggregates can be crashed by ultrasonic processing, engendering lower molecular PrP(Sc) polymers.

Animals ; Blotting, Western ; Brain Chemistry ; Cricetinae ; Image Processing, Computer-Assisted ; PrPSc Proteins ; analysis ; Scrapie ; metabolism ; Sonication