Objective:To investigate the quantitative expression of Melanoma-associated antigen MAGE-E1 mRNA in glioma, and explore its potential for immunotherapy in glioma.Methods:To establish a quantitative real-time polymerase chain reaction ( qRT-PCR) method to quantitatively determine MAGE-E1 mRNA in glioma.A total of 47 human glioma and 14 normal brain tissue specimens were analyzed.MAGE-E1 mRNA was normalized to HPRT1, a housekeeping gene.MAGE-E1/HPRT1 was used to evaluate the expression level of MAGE-E1 mRNA.Results:High-level expression of MAGE-E1 was observed in 7.1%of normal brain and 66.0%glioma tissues,which shown significant difference with P<0.05.There was irrelevant between the expression of MAGE-E1 mRNA and clinicopathogical parameters ,such as gender ,age,histological subtype and grade.Conclusion:With higher expression of MAGE-E1 in glioma tissues,it might be a potential target for immunotherapy of glioma.

Animals ; Animals, Newborn ; Bone Marrow Transplantation ; Disease Models, Animal ; Humans ; Hypoxia, Brain ; physiopathology ; prevention & control ; Rats ; Rats, Sprague-Dawley ; Stromal Cells ; transplantation ; Transplants ; trends

Objective To investigate the clinical value of the probe melting curve analysis‐based PCR assay for the detection of three types of αT .Methods A total of 149 blood and prenatal archival DNA samples (6 of which were amniotic fluid samples)with three knownαT genes ,which included 63 carriers with Hb CS ,22 cases with Hb QS ,43 individuals with Hb WS and 1 double heter‐ozygote with Hb CS and Hb WS) as well as 20 samples with normalα‐globin gene sequence that had been confirmed by RBD com‐bined with DNA sequencing were selected to test the specificity of probe melting curve analysis by blind analysis .Results The probe melting curve analysis accurately detected 100 of the DNA samples previously characterized by S RBD combined with DNA sequencing .Conclusion Probe melting curve analysis‐based PCR assay for the detection ofαT is featured with rapidity and accuracy and can be applied to clinical and prenatal diagnosis .

Background Diabetes is one of the risk factors that leads to corneal neuropathy.Silent signal regulatory factor 1 (Sirt1) plays an important role in glucose metabolism,lipid metabolism,regulation of insulin secretion and is closely related to the nervous system disease.The relationship between Sirt1 and diabetic corneal neuropathy is not fully understood.Objective This study was to detect the expression of Sirtl in cornea and trigeminal ganglion with type 1 diabetes model mice and explore the association of Sirt1 expression with diabetic corneal neuropathy.Methods Eight C57BL/6-Ins2Akita/J male mice and eight wild-type C57BL/6 male mice in the same litter were selected as type 1 diabetes model group and control group,respectively.The mice of two groups were sacrificed in overdose anesthesia method at 12-month old.Histological examination of cornea and trigeminal ganglion was performed using hematoxylin and eosin staining.Expression and localization of Sift1 protein in cornea and trigeminal ganglion were detected using immunohistochemistry.Western blot assay and fluorescine quantitative PCR were respectively used to quantitatively analyze the expression of Sirt1 protein and Sirt1 mRNA.Results Trigeminal ganglion cells were uneven in size and shape with the loosened cellular arrangement and disorder neurofibrosis alignment,and the corneal epithelial cells were less in the C57BL/6-Ins2Akita/J mice,but the trigeminal ganglion cells and corneal epithelial cells were normal in wild-type C57BL/6 mice.Immunochemisty exhibited that Sirtl protein was expressed mainly in corneal epithelium and the expression of Sirtl protein was stronger in the C57BL/6 mice than that in C57BL/6-Ins2Akita/J mice.Fluorescine quantitative PCR assay showed that the gray scale value of Sirt1 mRNA in cornea in C57BL/6-Ins2Akita/J mice was lower than that of the wild-type C57BL/6 mice(0.56±0.03 vs.0.98±0.13) with significant difference (t =5.853,P =0.010).Western blot showed that the expression of Sirt1 protein in cornea was lower in C57BL/6-Ins2Akita/J mice than that of the wild-type C57BL/6 mice(0.78±0.017 vs.1.300±0.012) with significant difference(t =33.140,P =0.001).However,no significant differences were seen in the gray scale value of Sirt1 mRNA(2.45±0.18 vs.2.51±0.22) (t=0.587,P=0.599) and protein level(1.100±0.015 vs.1.110±0.017) (t =0.430,P=0.709) in trigeminal ganglion tissues between C57BL/6-Ins2Akita/J mice and wide-type C57BL/6 mice.Conclusions The corneal nerve and structure is abnormal in 12-month-old C57BL/6-Ins2Akita/J mouse.Sirt1 is involved in the pathogenesis of diabetic keratoneuropathy,suggesting that it may be a potential target.

Objective: To investigate the therapeutic effects of Tuina (Chinese therapeutic massage) in a knee osteoarthritis (KOA) rat model and its influence on proteins associated with the Wnt/β-catenin signaling pathway. Methods: A total of 32 specific-pathogen-free grade Sprague-Dawley rats were used. Eight rats were randomly selected as the control group (CG). The remaining 24 rats underwent intra-articular injections with 0.2 mL of 4％ papain to prepare the KOA rat models. After the model was established, the 24 rats were randomly and equally assigned to 3 groups, including a model group (MG), a Tuina group (TG), and a positive medicine group (PMG), with 8 rats in each group. The Lequesne score was applied to evaluate the success of model development. After the model was successfully established, the CG did not receive any intervention, and the TG was treated with local, clockwise annular Rou-Kneading around the knee joint with the thumbs. The pressure in the longitudinal direction was 3 N, and the frequency was designed to be 120-140 times/min for 15 min, followed by flexing the joint 10 times. The PMG was intragastrically administered with celecoxib [24 mg/(kg·bw)] every day. These interventions were performed once a day, 6 d per week, for a total of 4 weeks. After treatment, the Lequesne score was applied again to assess the severity of the KOA in the rats; hematoxylin-eosin (HE) staining and a mixture of equal volumes of aqueous solutions of safranin O-fast green were used to stain and observe the cartilage morphology and structure; the modified Mankin score was applied to evaluate the pathology; enzyme-linked immunosorbent assay method was used to quantify the C-telopeptide fragments of type Ⅱ collagen (CTX-Ⅱ) and cartilage oligomeric matrix protein (COMP); Western blotting was then applied to quantify Wnt4, β-catenin, matrix metalloproteinase 13 (MMP-13), and bone morphogenetic protein 2 (BMP-2) protein expression; immunohistochemistry was conducted to determine the percentage of collagen type X (ColX)-positive cells. Results: The Lequesne score of the TG and PMG was both lower than that of the MG (P<0.01); the HE staining, safranin O-fast green stained morphology and structure, and modified Mankin scores of the TG and the PMG were also better than those in the MG (P<0.01). Compared with the CG, the amounts of CTX-Ⅱ and COMP in the serum were significantly increased (P<0.01); the expression of Wnt4, β-catenin, MMP-13, and BMP-2 proteins in the cartilage tissue was significantly increased (P<0.01), and the percentage of ColX-positive chondrocytes was significantly increased (P<0.01) in the MG. In comparison with those in the MG, the amounts of CTX-Ⅱ and COMP were significantly decreased (P<0.01), the expression of Wnt4, β-catenin, MMP-13, and BMP-2 proteins was significantly decreased (P<0.01), and the percentage of ColX-positive chondrocytes was significantly decreased (P<0.01) in the TG and PMG. Compared with the PMG, the contents of CTX-Ⅱ and COMP and the expression of Wnt4, β-catenin, MMP-13, and BMP-2 proteins were decreased (P<0.05 or P<0.01); the percentage of ColX-positive chondrocytes was significantly decreased (P<0.01) in the TG. Conclusion: Tuina can relieve the degeneration of KOA, and the mechanism may be related to the down-regulation of the Wnt/β-catenin signaling pathway, the decrease in MMP-13 and BMP-2 protein expression, the reduction in chondrocyte extracellular matrix degradation, and slowing down the terminal cell differentiation.

OBJECTIVE:To provide reference for further formulation of the rational use of vancomycin. METHODS:Retro-spective analysis was conducted on the related information of discharged patients who intravenously used vancomycin from Jun. 2013 to Dec. 2014. RESULTS:178 patients were enrolled,with average age of 59.6 and 73.60% male,who were mainly with lung infectious(74.72%). Support examinations were sufficient before using of vancomycin. 66.29% patients were empirically giv-en vancomycin with pathogenic detection rate of 85.39%. 71.91% patients were conducted therapeutic drug monitoring with only 47.54% of first blood samples achieved the target range. CONCLUSIONS:Vancomycin application is generally rational in our hos-pital. However,issues like duration of empirical therapy,rational therapeutic monitoring,and individualized start dosing still need to be noticed.

Objective： To study the changes of water molecular diffusion in the ischemic region by using MR dephase technique and discuss the potential mechanism of the diffusion changes at early stage. Methods: Totally 43 cases were studied retrospectively. There were 10 cases whose MRI examinations were performed within 6 hours，12 cases from 7－24 hours，7 cases from 2－7 days, 8 cases from 8－14 days, 6 cases from 15 days to 2 months. The apparent diffusion coefficients in the ischemic region were calculated. Results: The ADCav in the grey matter was 8.61×10－4mm2*s－1. The ADCav decreased to (4.72×10－4±1.51×10－4) mm2*s－1 in ischemic region at superacute stage, ADCav ratio to contralateral corresponding region was 0.55±0.18, and ADCav increased to (5.68×10－4±1.22×10－4) mm2*s－1 during the time range of 2-7 days, （9.22×10－4±2.07×10－4） during the time range of 8－14 days, and approaching （26.42×10－4+9.65×10－4） mm2*s－1 during the time range of 2 months. The pearson product- moment correlation between the changes of diffusion value and time was sighificent (r=0.95, P<0.001). ADCv increased at superacute stage and decreased over time. Conclusion: The diffusion of water molecules in ischemic region decreased at superacute stage, and the ADC increased over time. The anisotropy increased at superacute stage and decreased as the course developed. DWI could detect ischemic lesion much earlier than CT and routine MR examination. DWI has great value in the diagnosis of superacute stroke. The mechanism of the diffusion changes at early stage may be the intracellular toxicity edema.

Hypoxia-inducible factor-1 (HIF-1) is a key transcription factor on hypoxia responses in mammalian tissues. HIF-1 plays as a positive factor in solid tumor and leads to hypoxia-driven responses that enhance its downstream gene expression for tumor growth and survival. LXY6099 was obtained by the structural modification and optimization of manassantin A (MA) as a high potent HIF-1 inhibitor. Antitumor activity of LXY6099 was observed in this study. LXY6099 with an IC50 value of 2.46 x 10(-10) mol x L(-1) showed more sensitive inhibition activity to HIF-1 than that of MA detected by reporter gene assay (> 100 folds). It showed strong inhibition on the growth of human solid tumor cell lines. Furthermore, LXY6099 exhibited significant antitumor activity against established human tumor xenografts in nu/nu mice with treatment of MX-1 breast cancer. Thus, LXY6099 as a novel HIF-1 inhibitor could be further developed into anti-cancer agents.
Animals ; Antineoplastic Agents ; pharmacology ; Breast Neoplasms ; metabolism ; Cell Line, Tumor ; Gene Expression Regulation, Neoplastic ; Heterografts ; Humans ; Hypoxia-Inducible Factor 1 ; metabolism ; Lignans ; pharmacology ; Mice, Nude

Objective To analyze the dynamic evolution of brain MRI in patients with mitochondrial myopathy,encephalopathy,lactic acidosis,and stroke-like episodes (MELAS) syndrome.Methods A retrospective study was performed on 58 MELAS cases with pathologically and (or) molecularly confirmed diagnosis.MRI were repeated within 60 days after the onset of stroke-like episodes (SLE) and the evolution changes of cerebral lesions were accessed.Brain atrophy index (BAI) was calculated in the remission stage from 31 patients with MELAS,and the correlation between BAI,age and disease duration was analyzed.Results The proportion of lesions expansion,migration and shrink within 30 days after the onset of SLE was 64.1％ (25/39),10.2％ (4/39),17.9％ (7/39),respectively,and 13％ (3/23),21.7％ (5/23),56.5％ (13/23),between 30-60 days after the onset of SLE respectively.In the recovery stage of SLE,the BAI in 31 patients with MELAS was 15.2％ ±2.8％.The correlation coefficient between BAI and the age,total disease course and duration of encephalopathy was 0.329 (P =0.043),0.405 (P =0.012) and 0.649 (P =0.000).Conclusions Brain atrophy in the studied MELAS patients gradually develops and strokelike lesions shrink with progression of the disease.However,the migration of lesions is persistent.

Objective To establish protein fingerprints of common bacteria in clinics and to lay a foundation for rapid identification of bacteria.Methods Strains of Escherichia coli,Klebsiella pneumoniae,Pseudomonas aeruginosa and Staphylococcus aureus were detected by surface enhanced laser desorption and ionization time-of-flight mass spectrometry (SELDI-TOF MS).Stable expression protein peaks were screened and the data was input into the self-constructed Fingerwave software for identification of target bacteria by protein fingerprint comparison.Two hundred and fifty-six clinical isolates,including E.coli,K.pneumoniae,P.aeruginosa and S.aureus were detected and the data was compared with constructed database to evaluate its diagnostic value.Results The protein fingerprints including four common bacteia was used to identify the target bacteria with identification rate of 93.1％ (54/58) for E.coli,87.2％ (75/86) for K.pneumoniae,96.2％ (60/63) for P.aeruginosa and 96.2％ (51/53) for S.aureus,respectively.Conclusion Common bacteria can be rapidly identified by using the protein fingerprint comparison,which provides a powerful tool for bacterial identification.