Objective To investigate the clinical and pathological characteristics of normotensive pheochromocytomas ( NP) . Methods This retrospective study included 97 patients with a pathological diagnosis of pheochromocytoma at the Drum Tower Hospital Affiliated to Nanjing University Medical School during January 2004 to December 2013. All available clinical, biochemical, and radiological records were reviewed in these patients who were then categorized into hypertensive pheochromocytomas (HP) (n=64) and NP (n=33) groups. 97 cases of Adrenal Gland Scale Score of pheochromocytoma were examined, including tissue microscopic pathology assessment, ki67 and phenylethanolamine-N-methyltransferase ( PNMT ) immunohistochemistry and catecholamine type. Biochemical examinations of 95 subjects with primary hypertension ( PH) were recorded for comparative study. Results The patients with NP showed lower proportion of clinical triad than HP, inapparent metabolic disorders, and lower urinary catecholamine levels than HP, but showed higher results than primary hypertension. The weight of tumor was positively correlated with 24 hour urinary norepinephrine level in patients with HP(Y=1. 376+0. 653X,R2=0. 118, P=0. 028), but not in patients with NP;and the size or diameter of the tumor was negatively correlated with PNMT immunohistochemistry in patients with NP(Y=0. 940-0. 356X, R2=0. 494, P=0. 005), but not in patients with HP, indicating that NP may be misdiagnosed clinically. Conclusion Patients with NP have distinct clinical, biochemical, and pathological phenotypes; the phenotypic changes are closely related with the expression levels of catecholamine pathway products during the occurrence and development of the tumors.

OBJECTIVETo investigate the effects of the new antiallergic agent N-(pyridin-4-yl)-(indol-3-yl) acetamide-45 (acetamide-45) on electric field stimulation (EFS)-and methacholine-induced contraction of airway smooth muscle in vitro.

METHODSContractions were induced by EFS in isolated trachea and bronchus of rats or by cumulative methacholine concentrations in isolated trachea of guinea pigs. Changes in isometric force of isolated airway smooth muscle were measured by force transducers and recorded on a multi-channel polygraph recorder.

RESULTSAcetamide-45 inhibited the contraction induced by EFS in isolated rat airway. The IC50 was 10.74 (95% CI 8.87-13.00) micromol.L(-1) and 18.83 (95% CI 14.57-24.33) micromol.L(-1) in tracheae and bronchi, respectively. Acetamide-45 also inhibited methacholine-induced contractile response of isolated guinea pig trachea in a concentration-dependent manner. At concentrations of 3, 10, 30 micromol.L(-1) acetamide-45 significantly decreased maximal contractile response of methacholine by 24.6%-43.2% and increased EC50 of methacholine by 3.1-to 21.4-fold.

CONCLUSIONAcetamide-45 inhibits EFS-or methacholine-induced contraction of isolated airway smooth muscle, and these effects might be non-specific inhibition on cholinergic receptor.

Acetamides ; pharmacology ; Animals ; Depression, Chemical ; Electric Stimulation ; Guinea Pigs ; In Vitro Techniques ; Male ; Methacholine Chloride ; pharmacology ; Muscle Contraction ; drug effects ; Muscle, Smooth ; drug effects ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Trachea ; drug effects

BACKGROUNDTo determine the antigenicity of recombinant hepatitis E virus ORF2 (rHEV ORF2) protein expressed in Pichia pastoris (P. pastoris).

METHODSBy using the rHEV ORF2 protein from E.coli as control, an indirect ELISA was adopted to identify the sensitivity, specificity and stability of rHEV ORF2 protein from P. pastoris in detection of HEV IgM and IgG antibody in sera from patients with hepatitis E. The reactivity of the rHEV ORF2 against 5 HEV ORF2 monoclonal antibodies (McAbs) was also tested.

RESULTSThe minimum concentration of coated antigen with which HEV IgG could be detected was 12.5 ng/ml, while the highest serum dilution to detect both IgM and IgG antibodies against HEV was 1:5 120. No cross-reaction was found with sera from patients with any other types of hepatitis. The 37 degree C acceleration test showed that the rORF2 was highly stable within 12 months at 4 degrees C. The 5 HEV ORF2 McAbs showed better reaction with the rORF2 from P. pastoris, especially that 4B2, 2E2, whose reaction against the rORF2 were 125 and 25 times respectively higher than that of rORF2 from E.Coli.

CONCLUSIONThere may be more extensive conformational epitopes in the rHEV ORF2 from P. pastoris. The excellent antigenicity, sensitivity and stability suggest that it can be served as a new candidate antigen for the development of diagnostic reagents of hepatitis E.

Gene Expression ; Hepatitis Antibodies ; blood ; Hepatitis Antigens ; genetics ; immunology ; Hepatitis E ; immunology ; Hepatitis E virus ; genetics ; immunology ; Humans ; Pichia ; genetics ; metabolism ; Recombinant Proteins ; genetics ; immunology ; Viral Proteins ; genetics ; immunology

OBJECTIVETo verify the efficacy of Jianpi-xingniao needling therapy on prevention and treatment of motion sickness.

METHODSSixty volunteers of motion sickness were randomized into an acupuncture group and a delayed acupuncture group, 30 cases in each one. In the acupuncture group, acupuncture was given at Baihui (GV 20), Sishencong (EX-HN 1), Neiguan (PC 6), Zusanli (ST 36) and Hegu (LI 4). The needles were retained for 20 min. The treatment was given twice a week and 10 treatments were required. In the delayed acupuncture group, acupuncture was postponed, meaning no acupuncture during observation stage. Graybel scale was adopted to observe the score of symptoms and physical signs of the subjects of motion sickness before and after intervention. The efficacy was compared between the two groups.

RESULTSTwenty-five cases in the acupuncture group and 22 cases in the delayed acupuncture group were included in the statistical analysis. The score of symptoms and physical signs of motion sickness was reduced significantly after treatment in the acupuncture group as compared with that before treatment (10.12 +/- 3.37 vs 0.92 +/- 0.40, P < 0.05). The score in the acupuncture group was lower apparently than that in the delayed acupuncture group (0.92 +/- 0.40 vs 9.86 +/- 2.53, P < 0.05). The difference was not significant before and after treatment in the self-comparison of the delayed acupuncture group (P > 0.05). The total effective rate was 96.0% (24/25) in the acupuncture group, which was significantly better than 0.0% (0/22) in the delayed acupuncture group (P < 0.01).

CONCLUSIONJianpixingniao needling therapy relieves the symptoms of motion sickness in the patients and achieves a better clinical efficacy.

Acupuncture Points ; Acupuncture Therapy ; Adult ; Female ; Humans ; Male ; Motion Sickness ; therapy ; Treatment Outcome ; Young Adult

OBJECTIVETo investigate the prognostic predictors of nasal NK/T cell lymphoma.

METHODSThe clinicopathologic feature data of 61 patients with nasal NK/T cell lymphoma proven by pathological examination from Jan. 1997 to Jan. 2005 were collected. Expression of survivin, CD44, nm23, p53, Ki-67, MDR-1 and CD95 was detected by immunohistochemical staining in 30 patients with available histologic specimens. The correlation between these factors and prognosis were analyzed.

RESULTSIn univariate analysis, performance status, LDH level, clinical stage, initial treatment response, CD56, Ki-67 and CD95 were found to be the prognostic factors associated with time to progression (TTP) in nasal NK/T cell lymphoma, while the performance status, B symptoms, LDH level, initial treatment response, Ki-67 and CD95 were demonstrated as prognostic factors related to overall survival. In multivariate analysis, clinical stage, initial treatment response and performance status were independent prognostic factors for TTP, while the latter two factors were independent prognostic factors of overall survival.

CONCLUSIONClinical stage and initial treatment response, and performance status are found to be independent prognostic factors for TTP, whereas the latter two factors are demonstrated as independent prognostic factors of the overall survival. Overexpression of Ki-67 may be an unfavorable prognostic factor, but overexpression of CD95 may be a favorable one.

Adolescent ; Adult ; Aged ; Analysis of Variance ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Biomarkers, Tumor ; analysis ; Cyclophosphamide ; therapeutic use ; Doxorubicin ; therapeutic use ; Female ; Follow-Up Studies ; Humans ; Hyaluronan Receptors ; analysis ; Immunohistochemistry ; statistics & numerical data ; Inhibitor of Apoptosis Proteins ; Ki-67 Antigen ; analysis ; Killer Cells, Natural ; drug effects ; metabolism ; pathology ; Lymphoma, T-Cell ; drug therapy ; metabolism ; pathology ; Male ; Microtubule-Associated Proteins ; analysis ; Middle Aged ; Neoplasm Proteins ; analysis ; Neoplasm Staging ; Nose Neoplasms ; drug therapy ; metabolism ; pathology ; Prednisone ; therapeutic use ; Prognosis ; Proportional Hazards Models ; Vincristine ; therapeutic use ; fas Receptor ; analysis

OBJECTIVETo determine whether cysteinyl leukotriene receptor agonist LTD(4) and cysteinyl leukotriene receptor 1 (CysLT(1)) antagonist pranlukast affect the differentiation of human neuroblastoma SK-N-SH cells.

METHODSSK-N-SH cell morphological changes induced by LTD(4), pranlukast and LTD(4) + pranlukast were observed with retinoid acid (RA) as the positive control. The expressions of CysLT(1) and CysLT(2) receptors were detected by immunoblotting analysis, and the expression of microtubule-associated protein-2 (MAP-2), a neuron marker, was detected by fluorescent immunostaining.

RESULTThe immunoblotting results showed that SK-N-SH cells expressed CysLT(1) receptor moderately, and CysLT(2) receptor highly. The morphological results showed that RA, pranlukast and LTD(4) + pranlukast induced the compaction of the cell bodies and the outgrowth of neurites, while LTD(4) had no significant effect. The immunostaining results showed that MAP-2 was distributed in the cell bodies in control or pranlukast-treated cells; it was distributed in cell bodies and neuritis in RA-treated cells. Pranlukast increased the numbers of MAP-2-positive cells.

CONCLUSIONThe CysLT(1)receptor antagonist pranlukast modulates the differentiation of SK-N-SH cells.

Cell Differentiation ; drug effects ; Cell Line, Tumor ; Chromones ; pharmacology ; Humans ; Immunoblotting ; Immunohistochemistry ; Leukotriene Antagonists ; pharmacology ; Leukotriene D4 ; pharmacology ; Membrane Proteins ; metabolism ; Microtubule-Associated Proteins ; metabolism ; Neuroblastoma ; metabolism ; pathology ; Receptors, Leukotriene ; metabolism

OBJECTIVETo investigate the relationship between phospholipase D (PLD) activation and neutrophil priming induced by cardiopulmonary bypass (CPB), and try to clarify whether CPB-induced systemic inflammatory response can be attenuated by inhibiting neutrophilic PLD activation.

METHODSNeutrophils were isolated from arterial blood of 8 patients undergoing valve replacement before operation and 30 min after initiation of CPB respectively. Both the preoperative and CPB-stirred neutrophils were subdivided into 5 groups by receiving different experimental interventions: (1) bacterial lipopolysaccharide (LPS, 10 ng x ml(-1)), (2) N-formylmethionylphenylalanine (fMLP, 1 micromol x L(-1)), (3) LPS+fMLP, (4) 1-butanol (0.5%)+LPS+fMLP, (5) vehicle. Elastase and myeloperoxidase (MPO) release was measured for the parameters of neutrophil activation, neutrophil PLD activity was determined by quantitation of choline produced from the stable product of phosphatidylcholine catalyzed by PLD.

RESULTS(1) Preoperative neutrophils treated with LPS+fMLP presented significantly higher PLD activity (13.48+/-2.61 nmol choline x h(-1) x mg(-1)) and released more elastase and MPO than cells treated with vehicle (PLD activity 3.70+/-0.49 nmol choline x h(-1) x mg(-1)), P<0.01), LPS (P<0.01) and fMLP respectively. In 1-butanol+LPS+fMLP group, PLD activity of preoperative neutrophils was lower than that in LPS+fMLP group (P<0.01), besides the release of elastase and MPO decreased sharply below both LPS+fMLP and fMLP groups (P<0.01). In LPS group, PLD activity was higher (P<0.01), while elastase and MPO release did not differ from control. fMLP group presented PLD activity, elastase and MPO release higher than control (P<0.01); nevertheless, lower than LPS+fMLP group (P<0.01). (2) CPB-stirred neutrophils presented prominent PLD activity increment, and even the control level was 3.59-fold of the pre-operative control (P<0.01). PLD activity in LPS+fMLP group was higher than that in other groups. Notably, PLD activity was even nonstatistically lower in 1-butanol+LPS+fMLP group than that in LPS or fMLP group. CPB-stirred neutrophils in LPS+fMLP group released more elastase and MPO than control, LPS, and 1-butanol+LPS+fMLP groups did (P<0.01); however, neither of the release was statistically different from that of fMLP group.

CONCLUSIONSCardiopulmonary bypass enables neutrophil priming accompanied with significant increase in PLD activity. Inhibition of neutrophil PLD activation attenuates its priming and may alleviate CPB-induced systemic inflammatory reaction.

Adolescent ; Adult ; Cardiopulmonary Bypass ; adverse effects ; Female ; Humans ; Male ; Middle Aged ; Neutrophil Activation ; physiology ; Phospholipase D ; pharmacology ; Systemic Inflammatory Response Syndrome ; etiology ; physiopathology

OBJECTIVETo investigate whether or not lipopolysaccharide (LPS) and ovalbumin (OA) prime rat peripheral leukocytes, the effect of sensitization on priming and the role of phospholipase D in priming.

METHODSThe peripheral leukocytes were separated and purified from sensitized or unsensitized rats. LPS or OA was used as a priming agent and formylmethionylphenylalanine (fMLP) as an activating agent. Degradation of leukocyte was determined by measurement of elastase release and myeloperoxidase (MPO) activity. Phospholipase D (PLD) activity was assayed by the generation of choline,which was measured by choline-oxidase-catalyzed formation of H(2)O(2) and Trinder reaction.

RESULTCompared with cells treated by fMLP alone,leukocytes from unsensitized rat challenged with fMLP after incubated with LPS released more elastase and MPO (P<0.05). But there was no significant difference between leukocytes challenged with fMLP after incubated with OA and fMLP treated alone. In sensitized rat,there was no difference between leukocytes challenged with fMLP after incubated with LPS and fMLP treated alone. But leukocytes challenged with fMLP after incubated with OA released significantly more elastase and MPO than fMLP treated alone (P<0.05). A significant correlation was obtained between the release of elastase and PLD activity (r(s)=0.51,P<0.01), and also between the release of MPO and PLD activity (r(s)=0.73,P<0.01) in unsensitized rat. In sensitized rat, it was 0.48 (P<0.01) and 0.37 (P<0.05) respectively.

CONCLUSION(1) LPS primes peripheral leukocytes from unsensitized rats; (2) OA primes peripheral leukocytes from actively sensitized rats; (3) PLD plays a role in priming of rat peripheral leukocytes.

Animals ; Leukocyte Elastase ; secretion ; Leukocytes ; drug effects ; enzymology ; Lipopolysaccharides ; pharmacology ; Male ; N-Formylmethionine Leucyl-Phenylalanine ; pharmacology ; Ovalbumin ; immunology ; Peroxidase ; blood ; Phospholipase D ; physiology ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo prepare and purify recombinant human NAMPT and NAMPT (H247A) proteins and to detect their enzymatic activity.

METHODSUsing pcDNA3.1-hnampt as template, full-length hnampt was sub-cloned into pET-11a(+) plasmid. The hnampt (H247A) mutant was obtained by site-directed mutagenesis. The plasmids were introduced in Escherichia coli BL21star for protein expression. The recombined NAMPT and NAMPT (H247A) proteins were purified by flowing through nickel column and size-exclusion column. The target proteins were confirmed by SDS-PAGE and mass spectrometry detection. The enzymatic activities of recombinant proteins were assessed by solution NMR.

RESULTThe DNA sequences showed that hnampt (wild type) and hnampt (H247A) (mutation) were cloned into pET-11a(+). The recombinant proteins were expressed in Escherichia coli BL21star in soluble form. The purified protein was confirmed to be NAMPT with a molecular weight of 56 KD. The enzyme activity of NAMPT (H247A) was dramatically decreased compared to wild-type NAMPT.

CONCLUSIONThe recombinant hNAMPT and hNAMPT (H247A) proteins have been successful prepared and purified. The H247A mutation dramatically decreases the enzymatic activity of NAMPT.

Base Sequence ; Cytokines ; genetics ; isolation & purification ; metabolism ; Escherichia coli ; genetics ; Genetic Vectors ; Humans ; Molecular Sequence Data ; Mutagenesis, Site-Directed ; Mutation ; Nicotinamide Phosphoribosyltransferase ; genetics ; isolation & purification ; metabolism ; Plasmids ; genetics ; Recombinant Proteins ; genetics ; metabolism ; Transformation, Bacterial