Objective To study the effect of transfection with antisense DNMT3b gene eukaryotic expression plasmid on the growth of human cholangiocarcinoma cell line QBC939,and to explore the role of DNMT3b in the cholangiocarcinoma tumorigenesis.Methods The constructed antisense DNMT3b gene eukaryotic expression plasmid was transfected into QBC939 cells using liposome.The expression level of DNMT3b protein was detected by Western blot after stable transfection.The growth curves of transfected cells and un-transfected cells were observed by MTT method respectively.The cell proliferation ability was also observed by the test of colony formation in soft agar.The alterations of the cell cycle and the apoptosis rate were detected by FCM.Results Following the transfection,the protein level of DNMT3b decreased significantly;transfection with antisense DNMT3b gene eukaryotic expression plasmid did not affect the cell growth curve of QBC939,and did not decrease the cell colony formation rate(P=0.717);transfection with antisense DNMT3b gene also did not result in cell cycle alterations or induce cell apoptosis(P=0.089).Conclusions Transfection with antisense DNMT3b gene eukaryotic expression plasmid can down-regulate the expression level of DNMT3b in QBC939.It can not affect the growth and proliferation of human cholangiocarcinoma cell line QBC939,nor alter the cell cycle and induce cell apoptosis.

Hypermethylation of the promoter region is an important mean for the transcriptional repression of a number of cancer-associated genes, and over-expression and/or increased activity of DNA methyltransferase are considered to be the main cause of promoter hypermethylation. In order to further explore the epigenetic mechanism of tumor suppressor gene RASSF1A inactivation, 5-aza-2'-deoxycytidine (5-Aza-CdR), a DNA methyltransferase inhibitor, was used to treat the human biliary tract carcinoma cell line QBC-939 at the concentration of 5 micromol/L for 24 h in this study. After the chemical intervention with 5-Aza-CdR, the methylation status in the promoter region of RASSF1A gene was detected by methylation specific PCR (MS-PCR), and the expression alteration of RASSF1A mRNA and protein were observed by RT-PCR and Western Blot respectively. Following the treatment with 5-Aza-CdR, methylation status in the promoter region of RASSF1A gene was reversed from methylation to unmethylation. A 280 bp DNA band which represented RASS1FA expression at transcriptional level and a 40 kDa (1 kDa=0.9921 ku) protein band which represented RASSF1A expression at protein level were detected by RT-PCR and Western Blot respectively in the experimental group cells and there were no corresponding bands in the control group cells. The experimental results suggest that 5-Aza-CdR can induce demethylation in the promoter region of RASSF1A. It can also reverse epigenetic transcriptional silencing caused by DNA methylation and induce the re-expression of RASSF1A in QBC-939. This study also suggest that the mechanism of RASSF1A inactivation is very closely related to the methylation of the promoter region, which may provide a new epigenetic understanding for tumor related gene inactivation and the pathogenesis of biliary tract carcinoma.

Objective To investigate the clinical effect of dorsal carpometacarpal reversed island flap with dorsal metacarpal nerve to reconstruct finger. Methods We designed the dorsal reverse carpal and metacarpal island flaps with nerve by using the adjacent two dorsal matacarpal arteries as blood-supply and applied the stand of bone and tendon in waste finger or the free iliac transplantation to reconstruct the every sensory finger. Results Eighten cases were survived completely, and the skin degloving injuries of the finger in 3 cases. The maximum of the flap was 9cm by 8cm. Patients were followed up 3 months to 2 years,7 weeks later pain sense of reconstructed finger was recovered. Sensation over S3 amounts to 89% of the digits. Two-point-discrimination of the digits was 5-10mm. After the operation, the reconstructed finger obtained good appearance, the sensory recovery approach normally. The patients have ability to fulfil daily activities. Conclusion This method has advantages as follow:simple and practical,high survive rate,low impairment,sensible and good appearance.

Objective To report the clinical effects of microsurgery in treatment of infected extremities after blood vessel prosthesis were transplanted.Methods From Jan.1998 to Dec.2008,8 cases of major vascular injuries in extremities were blood-supplied by cross bridge vascular anastomosis from uninjured extremities,including 4 cases of femoral artery and vein,2 cases of popliteal artery and vein,and 2 cases of brachial artery and vein. Results After 3 years of follow-up,blood circulation of infected extremities were reestablished in each of 8 cases,as well as function and appearance recovered.Conclusion The procedure of cross bridge vascular anastomosis from uninjured extremities may efficiently restitute the blood supply of the infected extremities after blood vessel prosthesis were transplanted,and decrease the rate of amputation.

Objective To evaluate the reliability of axial view of patella with the knee joint flexed at 60 and 90 degrees on imaging diagnosis of patellar subluxation with DR (digital radiography). Methods 60 cases confirmed patella subluxation were reviewed retrospectively. In addition to routine anteroposterior and lateral DR views, axial DR views of patella with the knee flexed at 60 degrees and 90 degrees were taken meanwhile. Results Of the 60 cases, 45 failed to provide any patellar problems, while the other 15 cases revealed suspicious imaging view of patella subluxation in their normal AP and lateral DR views. In the axial DR views of patella with the knee flexed at 60 degrees and 90 degrees, 50, 4 and 6 cases revealed subluxation, suspicious subluxation and normal alignment of patella. Conclusion Axial DR view of patella with the knee joint flexed at 60 and 90 degrees could provide helpful roentgenographic information in determining the existence of patella subluxation.

Hypermethylation of the promoter region is an important mean for the transcriptional repression of a number of cancer-associated genes, and over-expression and/or increased activity of DNA methyltransferase are considered to be the main cause of promoter hypermethylation. In order to further explore the epigenetic mechanism of tumor suppressor gene RASSF1A inactivation,5-aza-2'-deoxycytidine (5-Aza-CdR), a DNA methyltransferase inhibitor, was used to treat the human biliary tract carcinoma cell line QBC-939 at the concentration of 5 μmol/L for 24 h in this study. After the chemical intervention with 5-Aza-CdR, the methylation status in the promoter region of RASSF1A gene was detected by methylation specific PCR (MS-PCR), and the expression alteration of RASSF1A mRNA and protein were observed by RT-PCR and Western Blot respectively. Following the treatment with 5-Aza-CdR, methylaiton status in the promoter region of RASSF1A gene was reversed from methylation to unmethylation. A 280 bp DNA band which represented RASS1FA expression at transcriptional level and a 40 kDa (1kDa=0.9921 ku) protein band which represented RASSF1A expression at protein level were detected by RT-PCR and Western Blot respectively in the experimental group cells and there were no corresponding bands in the control group cells. The experimental results suggest that 5-Aza-CdR can induce demethylation in the promoter region of RASSF1A. It can also reverse epigenetic transcriptional silencing caused by DNA methylation and induce the re-expression of RASSF1A in QBC-939. This study also suggest that the mechanism of RASSF1A inactivation is very closely related to the methylation of the promoter region, which may provide a new epigenetic understanding for tumor related gene inactivation and the pathogenesis of biliary tract carcinoma.

Background: and Purpose Previous studies suggested increased visit-to-visit variability of total cholesterol (TC) is associated with stroke. This study aimed to investigate the associations of various lipids measurements variability and the risk of stroke and stroke type (ischemic and hemorrhagic stroke). Methods: Fifty-one thousand six hundred twenty participants in the Kailuan Study without history of myocardial infarction, stroke, and cancer who underwent three health examinations during 2006 to 2010 were followed for incident stroke. Variability in TC, triglycerides, high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C) measurements were measured using the coefficient of variation (CV), standard deviation (SD), variability independent of the mean (VIM), and average real variability (ARV). Results: During a median of 6.04 years of follow-up, 1,189 incident stroke (1,036 ischemic and 160 hemorrhagic stroke) occurred. In the multivariable-adjusted model, the hazard ratio (HR) comparing participants in the highest versus lowest quartile of CV of HDL-C were 1.21 (95% confidence interval [CI], 1.02 to 1.45; P for trend=0.013) for ischemic stroke. The highest quartile of CV of LDL-C was associated with 2.17-fold risk of hemorrhagic stroke (HR, 2.17; 95% CI, 1.25 to 3.75; P for trend=0.002) compared with the lowest quartile. We did not observe any significant association between TC and triglycerides variability with any of stroke. Consistent results were obtained when calculating variability index using SD, VIM, or ARV. Conclusions These findings suggest the high visit-to-visit HDL-C and LDL-C variability were associated with an increased incidence of ischemic and hemorrhagic stroke, respectively.

Objective To develop a nomogram based on pulmonary nodules preoperative CT signs and 3D quantitative parameters for predicting mediastinal lymph node metastases in patients with clinical stage ⅠA lung adenocarcinoma.Methods The imaging data of 164 patients who underwent preoperative CT scan and systematic lymph node dissection were analyzed retrospectively.Commercially available AI software was used to extract 3D quantitative parameters of pulmonary nodules automatically,and CT signs of pulmonary nodules were analyzed.Logistic regression was used to explore the role of these parameters in predicting pathological nodal involvement.A nomogram prediction model was established,then discrimination and calibration of the model were evaluated.Results Among 164 enrolled patients,19(11.6％)were tested positive for mediastinal lymph node metastases at pathology review.The nomogram incorporated spiculation,lobulation,the largest cross-sectional area,and carcinoembryonic antigen(CEA).The model showed great discrimination and calibration,with a C-index of 0.942[95％confidence interval(CI)0.923-0.961].The predicted value of the model fitted well with the actual observed value on the calibration curve.Conclusion The nomogram prediction model based on preoperative CT signs,3D quantitative parameters,and CEA can estimate the probability of mediastinal lymph node metastases in clinical stage ⅠA lung adenocarcinoma.This model may help with clinical decision-making and individualized evaluation.

BACKGROUND AND PURPOSE: The aim of this study was to identify the clinical characteristics and potential mechanisms relevant to pathological proteins in Parkinson's disease (PD) patients who experience fatigue. METHODS: PD patients (n=102) were evaluated using a fatigue severity scale and scales for motor and nonmotor symptoms. The levels of three pathological proteins-α-synuclein oligomer, β-amyloid (Aβ)(1-42), and tau-were measured in 102 cerebrospinal fluid (CSF) samples from these PD patients. Linear regression analyses were performed between fatigue score and the CSF levels of the above-listed pathological proteins in PD patients. RESULTS: The frequency of fatigue in the PD patients was 62.75%. The fatigue group had worse motor symptoms and anxiety, depression, and autonomic dysfunction. The CSF level of α-synuclein oligomer was higher and that of Aβ1-42 was lower in the fatigue group than in the non-fatigue group. In multiple linear regression analyses, fatigue severity was significantly and positively correlated with the α-synuclein oligomer level in the CSF of PD patients, after adjusting for confounders. CONCLUSIONS: PD patients experience a high frequency of fatigue. PD patients with fatigue have worse motor and part nonmotor symptoms. Fatigue in PD patients is associated with an increased α-synuclein oligomer level in the CSF.
Anxiety ; Cerebrospinal Fluid ; Depression ; Fatigue* ; Humans ; Linear Models ; Parkinson Disease* ; Weights and Measures