Objective To build a more perfect serum protein ifngerprint models for early diagnosis of ganglioneuroblas-toma (GNB) in children. Methods Thirty children with GNB and 30 normal control children were recruited. Serum samples were collected. Nonspeciifc serum protein was detected and studied by MB-WCX processing, SELDI-TOF-MS mass spectrom-etry system and MALDI-TOF/TOF platform. Results Through the SELDL-TOX-MS processing, a peak at 5920 m/z protein markers, and the expression of the markers was high in GNB children (6180.6±2328), compared with normal control children (419.1±493.3), the difference was statistically signiifcant (P<0.05);MALDI-TOF/TOF platform showed that the protein with a peak at 5920 m/z is identiifed as ApoC-Ⅲ. Conclusions m/z peak of 5920 protein is suggested as speciifc biomarker of GNB in children, can provide signiifcant reference for early diagnosis of ganglioneuroblastoma, and prognostic monitoring.

Objective:To investigate the type and severity of skin adverse reactions induced by afatinib in the treatment of lung cancer, and to analyze their correlation with anti-lung cancer efficacy.Methods:A case-case-control study was conducted on lung cancer patients treated with afatinib in ZhongDa Hospital, Southeast University from December 2016 to January 2018. The type and severity of skin adverse reactions were evaluated in 76 patients with lung cancer based on the National Cancer Institute-Common Terminology Criteria for Adverse Events (NCI-CTCAE) Version 4.0, and these patients were divided into 3 groups according to the severity of skin lesions, including grade-0, -1, and -2/3 groups. The patients underwent chest computed tomography (CT) examination every 3 months, and the tumor response to afatinib was evaluated according to the Response Evaluation Criteria in Solid Tumors (RECIST) . Anti-lung cancer efficacy of afatinib was compared among the patients with different grades of skin lesions by using the Kruskal-wallis H rank sum test. Results:After treatment with afatinib, 44 of the 76 patients with lung cancer achieved stable condition or partial remission, and 32 experienced disease progression. Skin adverse reactions occurred in 69 patients, and manifested as acneiform lesions in 42 (55.3%) patients, paronychia in 35 (46.1%) , mucosal erosions in 30 (39%) , hair changes in 8 (10.5%) , and hand-foot syndrome in 6 (7.9%) . Improvement was achieved in 3, 7 and 34 cases in the grade-0, -1, and -2/3 groups ( n = 7, 19 and 50 respectively) , respectively. There was a significant difference in the response rate among the 3 groups ( χ2 = 6.117, P = 0.047) , and the response rate was significantly higher in the grade-1 and -2/3 groups than in the grade-0 group (both P < 0.001) , and higher in the grade-2/3 group than in the grade-1 group ( P < 0.001) . Conclusion:The treatment of lung cancer with afatinib can cause various types of skin lesions, such as acneiform lesions, paronychia, mucosal erosions, hair changes and hand-foot syndrome, and the higher the severity of the skin lesions, the more marked the anti-lung cancer efficacy of afatinib.

Objective To explore the association of Toll-like receptor 4 TLR4 and lipopolysaccharide receptor CD14 gene polymorphisms with Kawasaki disease (KD) susceptibility.Methods Three-color fluorescent staining flow-cytometry was used to detect the expression of TLR4 in peripheral blood white blood cell of 76 KD children and 118 healthy control group.The gene of TLR4 (-896A/G), (-1196C/T) and CD14 (-260C/T) polymorphisms was identified by polymerase chain reaction-restriction fragment length polymorphisms; and the relationship between genotype and KD was analyzed.Results 1.The values of mean fluorescence intensity (MFI) of TLR4 in peripheral blood white blood cell of the KD groups and the healthy control groups were 2.87?0.96, 10.55?4.87, 23.36?8.28 and 3.26?0.65, 7.55?1.21, 25.41?6.97, respectively; There was a gradual increase of these values on lymphocyte, neutrophilic leukocyte and mononuclear cell in both groups.2.(-896A/G), (-1196C/T) polymorphisms of TLR4 gene were not found in both groups.3.The frequency of each genotype of CD14 gene (-260C/T) was 35.5%CC, 30.3%CT, 34.2%TT in KD group and 38.1%CC, 47.5%CT, 14.4%TT in healthy control group.The frequency of each genotype was significantly different in 2 groups(?2=11.62 P

OBJECTIVETo test whether postconditioning could inhibit the expression of phospho-JNK (P-JNK) mitogen activated protein kinase (MAPK) and study its relation to apoptosis of cardiocyte.

METHODSSixty rats were randomly divided into six groups: sham, reperfusion injury (R/I), postconditioning (Post), SP600125 (I_JNK), anisomycin and postconditioning (Ani+Post) and anisomycin (Ani) groups. After acute myocardial infarction was induced in rats, placebo solution (DMSO), SP600125 (6 mg/kg) or anisomycin (2 mg/kg) was injected through jugular vein 5 min before reperfusion; 6 h later 3 rats of each group were executed and the hearts were separated to measure the signaling molecules (phospho-JNK, TNF alpha, Caspase-8, Bcl-2/Bax, cytochrome-c). Twenty-two hours later hemodynamic data were measured in the left rats, and then blood samples were taken to determine serum markers of cardiac damage, and hearts were separated to measure the infarction area and cardiocyte apoptosis.

RESULTPostconditioning improved +/-DP/DTmax of left ventricle, limited infarct area, relieved apoptosis and necrosis of cardiocytes, and inhibited the expression of P-JNK (1.12 +/-0.21 Compared with 1.90 +/-0.32, P<0.05). At the same time the levels of TNFalpha Caspase-8, Bax and Cyt-c were lower in Post group than those in R/I group, but Bcl-2 expression levels were higher. I_JNK group presented the similar protection effect of postconditioning [TUNEL index: (6.23 +/-2.43)% Compared with (18.22 +/-5.10)%, P<0.05; Infarct area: (23.44 +/-6.34)% Compared with (42.31 +/-8.21)%, P<0.05]. On the other hand, Ani+Post group partially lost cardioprotection effect [TUNEL index: (14.12 +/-2.00)% Compared with (18.22 +/-5.10)%,P>0.05; Infarct area: (35.27 +/-5.28)% Compared with (42.31+/-8.21)%,P>0.05], because of the activation of JNK MAPK.

CONCLUSIONPostconditioning can inhibit phosphorylation of JNK MAPK, which attenuates cardiocyte apoptosis by both extrinsic and mitochondria pathway.

Animals ; Apoptosis ; drug effects ; Ischemic Preconditioning, Myocardial ; JNK Mitogen-Activated Protein Kinases ; metabolism ; pharmacology ; Male ; Myocardial Infarction ; enzymology ; pathology ; therapy ; Myocardial Reperfusion Injury ; prevention & control ; Myocytes, Cardiac ; enzymology ; pathology ; Random Allocation ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo simulate the surgical approaches for intracranial aneurysms using three-dimensional CT angiography (3D-CTA) and assess the value of 3D-CTA in early microneurosurgery for ruptured intracranial aneurysms.

METHODSForty-eight patients with spontaneous subarachnoid hemorrhage due to ruptured intracranial aneurysm were confirmed by early operation. All the patients were classified according to Hunt-Hess, including 11 of grade I, 29 of grade II, and 8 of grade III. CTA was performed before the operation and surgical simulation was conducted. The preoperative findings on CTA and the intraoperative findings were compared and the clinical value of cerebral 3D-CTA was analyzed.

RESULTSPre-operative 3D-CTA clearly displayed the location, size and shape of the aneurysms, the axis direction of the aneurysm apex and the width of aneurysm neck. The spatial relation between the parent aneutysm artery, the aneurysm, the peripheral vessels and the bony structures were also demonstrated. These findings were basically consistent with the intraoperative findings. The Glasgow outcome score was 5 in 41 patients, 4 in 4 patients, 3 in 2 patients, and 2 in 1 patient upon discharge from the hospital.

CONCLUSIONSPreoperative 3D-CTA examination can simulate the surgery for ruptured aneurysms to help improve the surgical success rate.

Adult ; Aged ; Aneurysm, Ruptured ; diagnostic imaging ; surgery ; Cerebral Angiography ; methods ; Computer Simulation ; Female ; Humans ; Imaging, Three-Dimensional ; Intracranial Aneurysm ; diagnostic imaging ; surgery ; Male ; Middle Aged ; Radiography, Interventional ; Subarachnoid Hemorrhage ; diagnostic imaging ; etiology ; surgery ; Tomography, Spiral Computed

OBJECTIVETo explore the potential application of IFN-gamma enzyme-linked immunospot (ELISPOT) assay in the diagnosis of tuberculous epididymitis (TE) by comparing ELISPOT assay with the traditional purified protein derivative (PPD) tuberculin skin test.

METHODSWe examined 13 TE patients using an in-house ELISPOT kit, another 11 TE patients by PPD skin testing, and 57 healthy male volunteers by parallel test with both the methods.

RESULTSTwelve (92.3%) of the 13 TE cases were positive on ELISPOT assay, and 10 (90.9%) of the 11 TE cases positive on PPD skin test, with no statistically significant differences between the two groups (P > 0.05). Among the 57 healthy male volunteers, 8 (14.0%) were positive on ELISPOT, and 28 (49.1%) positive on PPD test, the latter significantly higher than the former (P < 0.001).

CONCLUSIONIn terms of sensitivity, ELISPOT assay is similar to PPD test in the examination of tuberculous epididymitis. As for specificity, ELISPOT assay seems better than PPD test in differentiating tuberculous epididymitis patients from healthy males.

Adult ; Case-Control Studies ; Enzyme-Linked Immunosorbent Assay ; Epididymitis ; diagnosis ; Humans ; Interferon-gamma ; Male ; Tuberculin ; Tuberculin Test ; Tuberculosis, Male Genital ; diagnosis

To study the genetic variation and evolutionary characteristics of H1N1 swine influenza virus, all the eight genes of LM were amplified by RT- PCR, cloned into pMD18-T vector and sequenced respectively. The results showed that neither insertion nor deletion was observed in nucleotides of LM. The amino acids sequence of cleavage site of HA is IPSIQSR decrease G, suggesting that LM did not have the molecular characteristics of high pathogen. HA had highly conservative N-glycosylation site at position 11, 23, 87 and 276 sites of HA1, and two more at position 154 and 213 sites of HA2. NA had highly conservative N-glycosylation site at position 58, 63, 68, 88, 146, and two more at position 44 and 235 sites, which might be one molecular characteristics of H1N1 subtype of SIV. The results of Bast showed HA gene had high homology to the strain of 'human-like' SIV (99%), while others had high homology to the 'classical' SIV. So it is inferred that HA of LM might originate from human-like linage swine influenza virus, while others might originate from 'classical' swine influenza virus.
Animals ; Cloning, Molecular ; Hemagglutinin Glycoproteins, Influenza Virus ; chemistry ; genetics ; Influenza A Virus, H1N1 Subtype ; classification ; genetics ; Neuraminidase ; chemistry ; genetics ; Phylogeny ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo screen and identify the serum specific protein markers of patients with gastric cancer by proteomics technology, and to provide more comprehensive serum protein fingerprint model for the early diagnosis of gastric cancer.

METHODSPreoperative and postoperative blood samples were collected from 60 gastric cancer patients. Mass spectrometry (SELDI-TOF-MS) technology was used to detect and screen serum specific proteins in gastric cancer patients(preoperative group, postoperative group, metastasis group), and the result was compared with normal control group. Gel electrophoresis(TRICINE SDS-OAGE) technology was applied in the separation and purification for those different protein. Matrix assisted laser desorption ionization tandem time-of-flight mass spectrometry (MALDI-TOF/TOF) technology was used in the identification for the proteins following separation and purification.

RESULTMass spectrometry data of preoperative group and normal group resulted in 15 specific m/z peak(P<0.01). SVM screened by a combination of the highest index model Youden get m/z peak at 6 449.1 protein markers. The protein expression of preoperative group was significantly higher than that of normal group(2 299.3±2 029.3 vs. 509.5±168.3, P<0.01). Mass spectrometry data of preoperative group and postoperative group resulted in 6 specific m/z peak(P<0.01). SVM screened by a combination of the highest Youden index model indentified get m/z peak at 6 449.2 protein markers. The protein expression of preoperative group was significantly higher than that of postoperative group(1 247.9±685.0 vs. 476.5±157.8, P<0.01). Mass spectrometry data of preoperative group and metastasis group resulted in 12 specific m/z peak (P<0.01). SVM screened by a combination of the highest Youden index model indentified get m/z peak at 6 448.9 protein markers. The protein expression of metastasis group was higher than that of preoperative group(1 506.9±1 036.5 vs. 649.7±621.0). MALDI-TOF/TOF identified that the protein with m/z peak at 6 449 was Apo CIII(.

CONCLUSIONApo CIII( may be the specific serum protein marker of gastric cancer, which may provide a more comprehensive serum protein fingerprint model for the early diagnosis of gastric cancer and a new way for further research.

Biomarkers, Tumor ; blood ; Blood Proteins ; analysis ; Humans ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; Stomach Neoplasms ; blood ; diagnosis

OBJECTIVEThe effects of various postconditioning algorithm on reperfusion injury and the role of mitochondrion pathway were investigated in a rat model of reperfusion/injury.

METHODSRats were divided into 5 groups: sham, reperfusion/injury (R/I group), reverse algorithm of postconditioning (R-Post, 30/10-25/15-15/25-10/30 s of reperfusion/re-occlusion), standard algorithm of postconditioning (S-Post, 4 cycles of 20/20 s of reperfusion/re-occlusion), and gradual algorithm of postconditioning (G-Post, 10/30-15/25-25/15-30/10 s of reperfusion/re-occlusion).

RESULTSThe levels of Bax, Cytochrome-c, Caspase-9, serum marker of myocardium and apoptosis index were significantly lower while the level of Bcl-2 was significantly higher in the three postconditioning groups than those in R/I group (all P < 0.05). The levels of Bax (0.35 +/- 0.10 vs. 0.50 +/- 0.02, P < 0.05), Cytochrome-c (0.66 +/- 0.16 vs. 1.68 +/- 0.22, P < 0.05), Caspase-9 (0.61 +/- 0.17 vs. 1.66 +/- 0.55, P < 0.05), serum marker of myocardium [CK: (251.00 +/- 45.16) U/L vs. (388.56 +/- 75.01) U/L, P < 0.05; CK-MB: (146.00 +/- 60.12) U/L vs. (291.16 +/- 52.41) U/L, P < 0.05] and apoptosis index [(4.32 +/- 1.16)% vs. (8.58 +/- 1.12)% , P < 0.05] were all significantly lower while Bcl-2 level (2.00 +/- 0.34 vs. 1.40 +/- 0.18, P < 0.05) was significantly higher in G-Post group than those in S-Post group. Moreover, above mentioned cardiac protective effects were significantly stronger in the G-Post group compared to R-Post group (all P < 0.05).

CONCLUSIONIn conclusion, gradual algorithm of postconditioning could attenuate reperfusion injury more significantly than standard algorithm, and mitochondrion pathway plays an important role in this cardioprotective process.

Algorithms ; Animals ; Caspase 9 ; metabolism ; Cytochromes c ; metabolism ; Ischemic Postconditioning ; Mitochondria ; metabolism ; Myocardial Reperfusion Injury ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Rats ; Rats, Sprague-Dawley ; bcl-2-Associated X Protein ; metabolism

OBJECTIVETo construct glypican-3 (GPC3)-green fluorescent protein eukaryotic expression vector pEGFP-c3-GPC3, and analyze the effect of GPC3 on the proliferation of human hepatoma cell line MHCC-97L.

METHODSThe eukaryotic expression vector pEGFP-c3-GPC3 was constructed with recombinant DNA technique and transfected into MHCC-97L cells via Lipofectamine 2000. The cells stably expressing GPC3 were screened by flow cytometry and G418. The mRNA expression of GPC3 was detected by RT-QPCR method, and the protein expression by Western blotting and fluorescence microscope. The effect of GPC3 gene on the growth of the cells was examined by MTT assay.

RESULTSRestriction endonuclease analysis and DNA sequencing verified correct construction of the recombinant plasmid. The green fluorescence was detected in the transfected MHCC-97L cells under fluorescence microscope. RT-QPCR and Western blotting both confirmed successful expression of GPC3 in MHCC-97L cells. The growth curve showed a significant acceleration of the proliferation of the transfected MHCC97-Lsol;GPC3 cells as compared with MHCC97-L and MHCC97-L/C3 cells (P<0.001).

CONCLUSIONWe have successfully constructed the eukaryotic expression vector pEGFR-c3-GPC3, which allows stable GPC3 expression in MHCC97-L/GPC3 cells. The upregulation of GPC3 expression can stimulate the growth of hepatoma cell line MHCC97-L in vitro.

Carcinoma, Hepatocellular ; pathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Genetic Vectors ; Glypicans ; pharmacology ; Green Fluorescent Proteins ; genetics ; Humans ; Liver Neoplasms ; pathology ; Plasmids ; Transfection