Objective To explore the effect of glutamine(Gln) on expression of nuclear factor kappa B(NF-?B) and heat shock protein 70(HSP70) in brain of young rats induced by lipopolysaccharide(LPS).Methods Ten days old Wistar rats were randomly divided into 3 groups by injection intraperitoneally different agonts,LPS group,normal saline control group(NS group) and Gln group(Gln 1.346 g/kg,1 hour before LPS).NF-?B and HSP70 distribution and expression in brain were deteted by immunohistochemistry.The levels of HSP70 in rats brain induced by LPS were detected by Western blot.SPSS 12.0 software was used.Results The nuclei of neuron in cerebral cortex in LPS group obviously cleared at 6 hours.The positive stain of nuclei in Gln group at 2 hours could not be seen.The stain of nuclei in cerebral cortex was weakened in LPS group at 6 hours by immunohistochemistry.HSP70 protein expression decreased with the measurement of Western blot,especially at 24 hours.HSP70 expression in LPS group was similar as that in NS group.The stain of nuclei in neuron in Gln group at 2 hours increased.It also showed the amount of protein expression increased in Western blot in group Gln at 2,6,12,24 hours(Pa

Catheter Ablation ; Consensus Development Conferences as Topic ; Humans ; Microsurgery ; Neoplasms ; therapy ; Radiotherapy Planning, Computer-Assisted ; Radiotherapy, Intensity-Modulated

The method of homogeneity evaluation for active pharmaceutical ingredient (API) spatial distribution in lyophilized product was investigated for the first time with confocal micro-Raman spectroscopy mapping, using pemetrexed disodium for injection as a model drug. Certain areas of the lyophilized product were scanned to obtain Raman spectra. The classical method ("peak clipping" method) was employed for mapping with characteristic Raman peaks of the API and the excipient. Due to the API being finely dispersed in the excipient in lyophilized products, the classical method cannot discriminate between the two ingredients making the distribution homogeneity difficult to evaluate. The "ratio of characteristic peak intensities" method was then utilized. Using this method, the relative intensity of the characteristic Raman peaks of the API to the excipient was applied for mapping and the relative content of API to excipient was calculated for a homogeneity evaluation of the drug distribution. The validation of this method showed a good linear relationship between the relative intensity and the relative content of API to excipient (r² > 0.99), and the precision and recovery were adequate for homogeneity evaluation of API by Raman spectroscopy mapping. Five products of pemetrexed disodium for injection from different manufacturers were tested through Raman maps applying this method and the histograms of relative Raman intensity were also plotted by frequency to help the homogeneity evaluation of drug distribution. The results showed that there were obvious differences in the drug distribution homogeneity from different products, where a more homogeneous API distribution was found in the brand product. This research provides a reliable method for the homogeneity evaluation of API distribution, which facilitates quality evaluation and process optimization of lyophilized products.

OBJECTIV: e To find the correlation between real best corrected visual acuity (BCVA) and testing results of microperimetry and visual evoked potential (VEP) and to explore a new method in recording BCVA in macular disease. METHODS: Sixty-two patients with macular disease (macular disease group, 62 eyes) and eighteen healthy volunteers (control group, 36 eyes) had BCVA, microperimetry and VEP recorded. RESULTS: (1) By microperimetry, the values of retinal mean sensitivity and fixation percentage in macular disease group were lower than that in control group. The bicurve ellipse area in macular disease group was higher than that in control group. By VEP, P100 amplitude under 0.5 cpd and 2 cpd in macular disease group were significantly higher than that in control group and the latency was prolonged (P < 0.05). (2) In macular disease group, BCVA had significant positive correlation with retinal mean sensitivity, bicurve ellipse area, macular central 2 degrees and 4 degrees fixation percentage, respectively (P < 0.05). There was a significant correlation between retinal mean sensitivity and P100 amplitude (P < 0.05). (3) Multiple linear regression equation was y = 0.053 x1+0.008 x3+3.897 (y was BCVA, while x1 was retinal mean sensitivity and x3 was P100 amplitude under 2 cpd). CONCLUSION Combined use of microperimetry and VEP is useful in the assessment of BCVA in macular disease.
Case-Control Studies ; Evoked Potentials, Visual/physiology* ; Eye ; Humans ; Macula Lutea/physiopathology* ; Retina ; Retinal Diseases/pathology* ; Tomography, Optical Coherence ; Visual Acuity/physiology* ; Visual Field Tests/methods*

The cDNA encoding human Vascular Endothelial Growth Factor 165 (VEGF165) was amplified using RT-PCR from human tonsil tissue and cloned into eukaryotic expression vector pcDNA3.1 (+). The recombinant plasmid pcDNA/V was transferred into 293 cells mediated by liposome and the cells stably expressing VEGF were selected under the pressure of G418. ELISA and Western blotting demonstrated that the eukaryotic expression vector pcDNA/V was successfully constructed and its corresponding protein could be expressed efficiently in vitro. Chick Charioallantoic Membrane (CAM) bioassay showed that recombinant protein has biological activity of hVEGF. Model rats with acute myocardial ischemia were used to further study the expression of VEGFin vivo. The model rats were divided randomly into three groups: control group, pcDNA3.1 (+) group and pcDNA/V group. 50microL naked plasmid DNA or saline was intramyocardially injected at three sites into the border zone of infarction. The hearts of rats were excised and fixed histologically, then the infarction sizes were studied by immunohistochemical staining and electron microscope after four weeks. Immunohistochemical staining for VEGF appeared to be negative in control and pcDNA3.1 (+) groups. In pcDNA/V group, myocardial cells in infarction border zone showed positive staining for VEGF in cytoplasm. Ultrastructural anaylsis showed that there were visible hyperplasia of vascular endothilium in pcDNA/V group. The control and pcDNA3.1 (+) groups showed less capillary hyperplasia. In this study, VEGF165 gene was successfully cloned and its protein expressed in vitro and in vivo was of bioactivity, which provides a basis for the further study of biological functions of human VEGF.
Animals ; Cell Line ; Chickens ; Chorioallantoic Membrane ; blood supply ; Disease Models, Animal ; Genetic Therapy ; Humans ; Male ; Myocardial Infarction ; metabolism ; pathology ; therapy ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Recombinant Proteins ; biosynthesis ; genetics ; therapeutic use ; Transfection ; Vascular Endothelial Growth Factor A ; biosynthesis ; genetics

OBJECTIVETo explore whether the constructed vector of short haprin in vivo can induce human glioma cell line BT325 to produce RNAi duplexes and reverse the expression of MDR1 gene.

METHODSThree 62nt oligonucleotide fragments (shRNA) were constructed according to GenBank MDR1 sequence and were cloned to the retrovirus-delivered vectors. After transfected these vectors directly into the human malignant glioma BT325 cells by lipofectamine 2000 with enhanced green fluorescence protein (EGFP) co-transfecting, the MDR1 gene silence effects were detected by the changing level of mRNA and P-glycoprotein including real time PCR (RT-PCR), Northern blot and Western blot analysis. To assess the multidrug resistance against adriamycin (ADR) and VCR, cell proliferation assays were performed by cell counting kit-8.

RESULTSThe RNAi plasmid vectors were constructed successfully. RT-PCR showed MDR1 mRNA was significantly reduced (P < 0.05). Northern blot analysis showed that the gene silence became most intense at 48 hours after transfection. Western blot analysis demonstrated that P-gp expression was reduced at different time to 12.9%, 30.3% and 4.8%, respectively. The chemosensitivity assays indicated that the transfected cells showed an enhanced sensitivity to ADR and VCR. Based on the value of IC(50), BT325 cells had significantly increased sensitivity to the drugs.

CONCLUSIONThe sequence specific RNAi can inhibit MDR1 mRNA and P-gp expression in the glioma cell line. It may reverse multidrug resistance phenotype, therefore, may provide promising therapeutic modalities in the treatment of human glioma.

ATP-Binding Cassette, Sub-Family B, Member 1 ; biosynthesis ; genetics ; Antibiotics, Antineoplastic ; pharmacology ; Antineoplastic Agents, Phytogenic ; pharmacology ; Cell Line, Tumor ; Doxorubicin ; pharmacology ; Drug Resistance, Multiple ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Glioma ; metabolism ; pathology ; Humans ; Plasmids ; RNA Interference ; RNA, Messenger ; biosynthesis ; genetics ; RNA, Small Interfering ; genetics ; pharmacology ; Transfection ; Vincristine ; pharmacology

BACKGROUND/AIMS: Recently, esophageal mean nocturnal baseline impedance (MNBI) and post-reflux swallow-induced peristaltic wave (PSPW) index have been proposed, which can increase the diagnostic role of multichannel intraluminal impedance and pH recording (MII/pH) for differentiating patients with heartburn. Therefore, our aim is to investigate the role of esophageal proximal MNBI, distal MNBI, and PSPW index in differentiating Chinese patients with heartburn. METHODS: Patients with heartburn from the Beijing Anzhen Hospital, who underwent upper gastrointestinal endoscopy and 24-hour MII/pH, were enrolled in this study. RESULTS: In all, 24 erosive esophagitis (EE), 46 non-erosive reflux disease (NERD), 52 reflux hypersensitivity (RH), and 78 functional heartburn (FH) patients were recruited. The respective median values for the EE, NERD, RH, and FH groups were as follows: proximal MNBI 1858.0, 2147.5, 2374.3, and 2329.0 Ω (P = 0.053); distal MNBI 1243.4, 1506.5, 2451.2, and 2477.3 Ω (P < 0.001); and PSPWI 15.0%, 25.0%, 25.0%, and 45.0% (P < 0.001). Spearman correlation analysis showed that distal MNBI and PSPW index were significantly negatively correlated with acid and bolus exposure time and acid reflux events. Receiver operating characteristic analyses showed that distal MNBI and PSPW index significantly discriminated FH from EE, NERD, and RH (P < 0.001), with cut-off values of 1890.6 Ω and 27.5% and areas under the curve of 0.721 and 0.779, respectively. CONCLUSION: Esophageal distal MNBI and PSPW index could increase the diagnostic role of MII/pH, especially for differentiating Chinese patients with heartburn.
Asian Continental Ancestry Group ; Beijing ; Electric Impedance ; Endoscopy, Gastrointestinal ; Esophagitis ; Esophagitis, Peptic ; Gastroesophageal Reflux ; Heartburn ; Humans ; Hydrogen-Ion Concentration ; Hypersensitivity ; ROC Curve

OBJECTIVETo explore the association between the genetic polymorphisms of mannose-binding protein (MBP) alleles and susceptibility to pulmonary tuberculosis.

METHODS125 pulmonary tuberculosis cases and 198 healthy controls were collected. A case-control study was conducted. Three structural gene mutations in exon 1 of MBP gene (codon 52, codon 54 and codon 57) were studied. Polymerase chain reaction with sequence-specific primers (PCR-SSP) was carried out in the polymorphism in MBP alleles. Information on related risk factors of tuberculosis was collected, using a pre-tested questionnaire. Univariate and multivariate logistic analyses were conducted with SPSS software package.

RESULTSThe frequencies of mutant heterozygote or homozygote of MBP-52, 54, 57 were 8.0%, 7.2% and 0.4% for cases and 5.3%, 4.3%, 0.5% for controls, respectively. The distribution of mutant genotypes of MBP did not show significant difference between tuberculosis patients and control by Mantel-Haenszel chi2 on sex. The univariate analysis demonstrated that body mass index, marital status, vaccinal vestige, bacillus of Calmette-Guerin vaccine immunization, contacted with pulmonary tuberculosis patients, familial traits were the risk factors of pulmonary tuberculosis. After adjusting those related environmental factors in the multivariate logistic analyses, the total MBP (MBP-52, MBP-54 and MBP-57) and MBP-52 heterozygote genotypes were significantly overrepresented in cases, with adjusted OR (95% CI) being 2.182 (1.058-4.499) and 2.574 (1.028-6.446).

CONCLUSIONTotal MBP and MBP-52 mutant genotypes might be associated with the susceptibility to pulmonary tuberculosis.

Case-Control Studies ; Genetic Predisposition to Disease ; Humans ; Mannose-Binding Lectin ; genetics ; Polymorphism, Genetic ; Tuberculosis, Pulmonary ; genetics

OBJECTIVETo investigate the effectiveness, safety and possible mechanism of recombinate human interleukin 11 (rhIL-11) in the treatment of chemotherapy-induced thrombocytopenia.

METHODSThirty-four patients (totally 76 cycles) with chemotherapy-induced thrombocytopenia received subcutaneous injection of rhIL-11 at the dose of 25 microg.kg(-1).d(-1) for 4 to 16 days. Serum IL-11 level was measured by ELISA, and IL-11 R alpha expression was detected by RT-PCR.

RESULTSThe mean baseline platelet count before chemotherapy was (135.0 +/- 54.3) x 10(9)/L for the 1st cycle and (259.4 +/- 64.5) x 10(9)/L for the 2nd cycle. The time to administer rhIL-11 was 7 to 16 days (median 12 days) in the 1st cycle and 4 to 10 days (median 6 days) in the 2nd, respectively (P < 0.05). The duration of post-chemotherapy platelet count below 50 x 10(9)/L was 7 to 13 days (median 10 days) for the 1st cycle and 3 to 8 days (median 5 days) for the 2nd, respectively (P < 0.05). Platelet count reached 300 x 10(9)/L or above in 30 chemotherapy cycles. The maximum platelet count was found to appear at D10 to D 17 (median D14), and negatively correlated with the pre-chemotherapy serum IL-11 level after administration of rhIL-11. Major adverse reactions included edema, headache, muscle and joint pain.

CONCLUSIONrhIL-11 is effective and safe for the treatment of chemotherapy-induced thrombocytopenia, with a relatively slow but sustained effect on the recovery of platelet count. Pre-chemotherpy serum IL-11 level might predict the efficacy of rhIL-11.

Adult ; Aged ; Aged, 80 and over ; Antineoplastic Combined Chemotherapy Protocols ; adverse effects ; therapeutic use ; Breast Neoplasms ; drug therapy ; Carcinoma, Non-Small-Cell Lung ; drug therapy ; Female ; Humans ; Injections, Subcutaneous ; Interleukin-11 ; administration & dosage ; adverse effects ; blood ; Lung Neoplasms ; drug therapy ; Male ; Middle Aged ; Platelet Count ; Recombinant Proteins ; administration & dosage ; adverse effects ; Thrombocytopenia ; chemically induced ; drug therapy ; Treatment Outcome

OBJECTIVETo investigate the clinicopathologic features of gastrointestinal stromal tumors (GISTs) and to identify reliable prognostic parameters.

METHODSFifty-nine GISTs were studied by immunostaining of CD117, CD34, SMA, desmin, S-100, bcl-2, and Ki-67. Histopathologic evaluations included tumor size, necrosis, histological growth patterns, mitotic activities and tumor lymphocytic infiltrate. The patients were clinically followed for 2 to 9 years. Univariate, multivariate and correlative statistical evaluations were used to analyze the data.

RESULTSAmong the 59 patients, 40 were alive and 15 died of their tumors at follow-up, the remaining 4 patients died of other causes. Pathological parameters that correlated with prognosis included tumor sizes of more than 5 cm, tumor tissue necrosis, mitotic cell count equal or higher than 5 per 50 high power field, Ki-67 labeling index (LI) equal or higher than 5% and intense bcl-2 immunostaining. Multivariate analysis showed that the mitotic count and Ki-67 LI were independent prognostic indicators. There was a correlation between mitotic count and Ki-67 LI.

CONCLUSIONSMitotic count and Ki-67 LI are the best predictors for a poor outcome of GIST after surgical treatment. Ki-67 immunostaining may substitute mitotic count as a useful prognostic parameter.

Adolescent ; Adult ; Aged ; Child ; Cyclin D1 ; metabolism ; Female ; Follow-Up Studies ; Gastrointestinal Stromal Tumors ; metabolism ; pathology ; Humans ; Ki-67 Antigen ; metabolism ; Male ; Middle Aged ; Prognosis ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Proto-Oncogene Proteins c-kit ; metabolism