Objective To detect the argyrophilic proteins in nucleolar organizer regions(Ag-NORs) that express rDNA and rRNA proliferation of T lymphocytes before chemotherapy and after complete remission(CR) in children with primary acute leukemia(AL).Methods The argyrophilic granules area of NOR/nuclear area(I.S%) of T lymphocytes was detected by image analysis system in peripheral blood of 42 patients before chemotherapy and after CR and 30 normal children.Results I.S% in the patients before chemotherapy(5.06%?1.36%) were significantly lower than those in the healthy donors(7.51%?1.06%)(t=8.238 P0.05).Conclusion These results suggest that decrease of Ag-NORs expresses the evidence for tumour induced suppression of immune function of T cells in children with AL prior to treatment.

OBJECTIVETo investigate the levels of pro-inflammatory cytokine IFN-gamma and anti-inflammatory cytokine TGF-beta1, in the expressed prostatic secretion (EPS) of men with chronic abacterial prostatitis and their clinical significance.

METHODSThe levels of IFN-gamma and TGF-beta1, in the EPS of 20 patients with inflammatory chronic pelvic pain syndrome (type III A), 20 patients with non-inflammatory chronic pelvic pain syndrome (type Ill B) and 10 healthy men were measured using enzyme-linked immunosorbent assay (ELISA). The results were analysed comparatively with NIH-chronic prostatitis symptom index (NIH-CPSI).

RESULTSIFN--gamma and TGF-beta1 levels were higher in III ([14.92 +/- 7. 85)], [8477.50 +/- 4612.45] ng/L) and III B ([13.74 +/- 5.96], [7946.50 +/- 5044.06] ng/L) prostatitis patients than those in the controls ([7.47 +/- 1.49], [2462.50 +/- 985.31] ng/L), P < 0.05 and P < 0.001 respectively. There was no statistically significant difference in cytokine levels between III A and Il B prostatitis patients. No correlation was found between NIH-CPSI and cytokine levels, r = 0.02, P = 0.86, r = 0.31, P = 0.76.

CONCLUSIONIFN-gamma and TGF-beta1, play a very important role in the etiology of chronic abacterial prostatitis and can be the objective parameters in the diagnosis of chronic abacterial prostatitis.

Adult ; Bodily Secretions ; chemistry ; Chronic Disease ; Enzyme-Linked Immunosorbent Assay ; Humans ; Interferon-gamma ; analysis ; Male ; Middle Aged ; Pelvic Pain ; diagnosis ; Prostate ; secretion ; Prostatitis ; diagnosis ; Transforming Growth Factor beta1 ; analysis

The aim of this study was to investigate the effect of platelet-rich plasma (PRP) on cavernous nerve (CN) regeneration and functional status in a nerve-crush rat model. Twenty-four Sprague-Dawley male rats were randomly divided into three equal groups: eight had a sham operation, eight underwent bilateral nerve crushing with no further intervention and eight underwent bilateral nerve crushing with an immediate application of PRP on the site of injury. Erectile function was assessed by CN electrostimulation at 3 months and nerve regeneration was assessed by toluidine blue staining of CN and nicotinamide adenine dinucleotide phosphate (NADPH)-diaphorase staining of penile tissue. Three months after surgery, in the group that underwent bilateral nerve crushing with no further intervention, the functional evaluation showed a lower mean maximal intracavernous pressure (ICP) and maximal ICP per mean arterial pressure (MAP) with CN stimulation than those in the sham group. In the group with an immediate application of PRP, the mean maximal ICP and maximal ICP/MAP were significantly higher than those in the injured control group. Histologically, the group with the application of PRP had more myelinated axons of CNs and more NADPH-diaphorase-positive nerve fibres than the injured control group but fewer than the sham group. These results show that the application of PRP to the site of CN-crush injury facilitates nerve regeneration and recovery of erectile function. Our research indicates that clinical application of PRP has potential repairing effect on CN and peripheral nerves.
Animals ; Disease Models, Animal ; Electric Stimulation ; Erectile Dysfunction ; pathology ; physiopathology ; therapy ; Male ; NADPH Dehydrogenase ; metabolism ; Nerve Regeneration ; physiology ; Penile Erection ; physiology ; Penis ; innervation ; Peripheral Nerve Injuries ; Peripheral Nerves ; metabolism ; pathology ; Platelet Transfusion ; Platelet-Rich Plasma ; Radiculopathy ; etiology ; pathology ; physiopathology ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo produce specific monoclonal antibody (mAb) against recombinant human erythropoietin (rHuEPO) for development of highly efficient methods for erythropoietin detection in biological fluids.

METHODSrHuEPO was covalently coupled with bovine serum albumin (BSA) and the conjugate was used to immunize mice to produce specific mAb against rHuEPO based on hybridoma technology. The obtained F3-mAb was characterized by enzyme-linked immunosorbent assay (ELISA), SDS-PAGE and Western blot.

RESULTSThe isotype of F3-mAb was found to be IgM with an affinity constant of 2.1 x 10(8) L/mol. The competitive ELISA using the obtained IgM showed a broader linear range and lower detection limit compared with previous work.

CONCLUSIONSThe modification of rHuEPO was proved to be successful in generating required specific mAb with high avidity to rHuEPO.

Animals ; Antibodies, Monoclonal ; biosynthesis ; immunology ; isolation & purification ; Antibody Affinity ; Dose-Response Relationship, Drug ; Enzyme-Linked Immunosorbent Assay ; Erythropoietin ; immunology ; Female ; Humans ; Immunoglobulin G ; immunology ; Immunoglobulin M ; immunology ; Mice ; Mice, Inbred BALB C ; Molecular Weight ; Recombinant Proteins

OBJECTIVETo investigate the role of ICC-like cells in bladder neuromodulation in rat urinary bladder.

METHODS14 SD rats and 1 guinea pig were sacrificed in this study. The ultra structural relationships among interstitial cells, nerves and detrusor smooth muscle cells (DSMCs) of urinary bladder were investigated by transmission electron microscopy (TEM). c-kit immunofluorescence was used to identify ICC-like cells in SD rat urinary bladder and the structural relationship between ICC-like cells and nerve terminals was studied by immunofluorescence (double-label).

RESULTSGap junction between ICC-like cells and DSMCs was confirmed by TEM. ICC-like cells were very close apposition with nerve terminals under TEM. ICC-like cells were identified to exist in sub-urothelium layer, along the longitude of smooth muscle bundles and among detrusor smooth muscle in SD rat urinary bladder by c-kit immunofluorescence. Double-labeled tissue with c-kit and PGP9.5 antibodies also showed that ICC-like cells were very close apposition with nerve terminals in SD rat bladder.

CONCLUSIONSMorphological study indicated that ICC-like cells in rat urinary bladder may play an important role in detrusor neuromodulation. Further study on function will be helpful for elucidating the mechanism of bladder neuromodulation clearly.

Animals ; Cells, Cultured ; Female ; Gap Junctions ; Guinea Pigs ; Male ; Muscle, Smooth ; innervation ; Myocytes, Smooth Muscle ; cytology ; ultrastructure ; Nerve Endings ; ultrastructure ; Rats ; Rats, Sprague-Dawley ; Urinary Bladder ; cytology ; innervation ; ultrastructure

This study was aimed to prepare the polyclonal antibody against the soluble proliferation-inducing ligand (sAPRIL) antigen and to investigate its effects in suppressing sAPRIL mediated lymphocyte proliferation. Mutated recombinant sAPRIL protein, which lacks biological activity but maintains immunogenicity, was used as antigen to immunize humanized SCID mice. Sera were obtained at 6 weeks after immunization. Indirect ELISA and Western blot were used to detect the antibody titer and specificity. The inhibition of polyclonal antibodies on Raji and Jurkat cell proliferation stimulated by sAPRIL was assessed by the MTT assay. The results showed that the mutant of sAPRIL could induce the production of polyclonal antibodies against human sAPRIL. Western blot and indirect ELISA analyses indicated that the anti-serum had higher specificity with a titer of 1:640. Functional analysis revealed that these polyclonal antibodies significantly inhibited the proliferation of Raji and Jurkat cell stimulated by sAPRIL (p < 0.05). It is concluded the polyclonal antibody against human sAPRIL is successfully prepared, which can inhibit the proliferation of Raji and Jurkat cells stimulated by sAPRIL in vitro.
Animals ; Antibodies ; genetics ; immunology ; pharmacology ; Antibody Specificity ; immunology ; Cell Proliferation ; drug effects ; Cloning, Molecular ; Humans ; Immune Sera ; analysis ; immunology ; Jurkat Cells ; Mice ; Tumor Necrosis Factor Ligand Superfamily Member 13 ; genetics ; immunology

OBJECTIVETo study the clinical features, treatment methods and outcome of solitary plasmacytoma of cervical spine.

METHODSFrom January 1995 to December 2007, the data of 23 cases with solitary plasmacytoma of cervical spine was analyzed. There were 16 males and 7 females (mean age 56 years, range: 32 - 76 years). Two cases underwent radiotherapy alone and 21 patients received surgery. According to WBB staging system, surgical procedures were defined as total or subtotal resection (6 cases), appendix resection (4 cases), sagittal resection (3 cases) and total spondylectomy (8 cases). All surgical cases were managed using an anterior approach, posterior approach or combined anterior and posterior approach. The cervical spinal reconstruction was achieved through anterior cervical titanium plate and titanium mesh cage filled with auto iliac graft or bone cement, or anterior and posterior combined instrumented fusion. All patients received radiotherapy as adjunctive therapy.

RESULTSFollow-up of the 23 cases lasted 24.0 - 143.0 months (mean: 64.7 months). Neck pains obviously improved, and nerve compression symptoms disappeared or improved after surgery. Neurological function improved by 1 - 2 grades based on Frankel grading system. All the internal fixations were fused well and stability of the cervical spine was fine and no spine instability could be seen in our series. The bone graft fusion rate was 100%. During the follow-up period, 6 surgical cases had local recurrence and finally progressed to multiple myeloma (MM) and 3 died. Two cases without surgical treatment progressed to MM in 1 year and 1.5 years after confirmed diagnosis. They were given systemic chemotherapy. The other 15 patients had disease-free survival and after surgery and adjunctive radiotherapy. Obvious abnormity were not found in such examinations as M protein, bone marrow aspiration and emission computed tomography or PET-CT examinations.

CONCLUSIONSSolitary plasmacytoma of cervical spine is rarely seen clinically. Surgery is recommended as the primary management for patients with overt bone destruction and spinal instability or neurological dysfunction. Tumor excision with adjunctive radiotherapy can obviously reduce local recurrences and lower the possibility of progression to MM. The patients with progression to MM should receive chemotherapy according to chemotherapy protocol while the prognosis is comparatively worse.

Adult ; Aged ; Cervical Vertebrae ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Plasmacytoma ; surgery ; Retrospective Studies ; Spinal Neoplasms ; surgery ; Treatment Outcome

OBJECTIVETo evaluate the effects of PTEN on invasive and migration ability of human ovarian cancer cell line A2780 and related mechanisms.

METHODSThe plasmid including WT-PTEN and mutant PTEN were transferred into A2780 cells. The invasive and migration ability were measured before and after transfection by transwell chamber and wound-healing assays. The expression of PTEN protein and related proteins in the cells were detected by Western blot analysis. Empty plasmid-transfected A2780 and normal A2780 cells were used as control (the different four groups were named as WT-PTEN/A2780, C124A-PTEN/A2780, GFP/A2780 and A2780).

RESULTSThe number of penetrating cells was significantly less in WT-PTEN/A2780 cells (24.3 ± 2.5) than those in C124A-PTEN/A2780, GFP/A2780 and A2780 cells (43.7 ± 3.8, 44.7 ± 2.1 and 45.0 ± 3.0) (P < 0.05). The migration distance was markedly shorter in WT-PTEN/A2780 cells (54.1 ± 3.7) than those in C124A-PTEN/A2780, GFP/A2780 and A2780 cells (78.7 ± 3.4, 78.1 ± 3.1 and 76.8 ± 3.5) (P < 0. 05).

CONCLUSIONSTransfection with PTEN may suppress the invasive and migration ability of ovarian cancer cell line A2780 depending on its phosphatase activity, and the suppressive effect may be due to the down-regulation of MMP-9 in the cancer cells.

Cell Line, Tumor ; Cell Movement ; Down-Regulation ; Female ; Genetic Vectors ; Humans ; Matrix Metalloproteinase 9 ; metabolism ; Mutation ; Neoplasm Invasiveness ; Ovarian Neoplasms ; metabolism ; pathology ; PTEN Phosphohydrolase ; genetics ; metabolism ; physiology ; Plasmids ; Transfection

In order to study the epidemiological characteristics of cytomegalovirus (CMV) infection in allogeneic hematopoietic stem cell transplantation (allo-HSCT) recipients by means of plasma real time quantitative polymerase chain reaction (RQ-PCR), 141 adult patients undergoing allo-HSCT between January 2008 and June 2010 were serially monitored by RQ-PCR for detecting CMV and guiding the preemptive therapy followed up to 180 days post-HSCT. The results showed that the incidence of CMV infection and CMV pneumonia was 81.5% and 2.9% respectively, which mainly occurred within 2 months post-HSCT. Single-therapy with ganciclovir (GCV) for 63 patients or foscarnet 6 patients was performed for preemptive therapy. The total efficacy was 87.8%, and the response patterns were different. CMV infection was more frequent in female patients (P = 0.044), and those with aGVHD (P = 0.043), using ATG or basiliximab in conditioning regimens (P = 0.049), as well as earlier in patients using ATG or basiliximab or those with aGVHD (P = 0.007; P = 0.000). The aGVHD, maximum load, positive times of CMV-DNA detection and therapy duration all correlated with the efficacy (P < 0.05). It is concluded that the incidence of CMV infection is still high after HSCT. Plasma RQ-PCR assay for CMV-DNA shows a strong correlation with the clinical outcome of CMV infection, which is useful and suitable for management of CMV infection in HSCT.
Adolescent ; Adult ; Antiviral Agents ; therapeutic use ; Child ; Cytomegalovirus ; Cytomegalovirus Infections ; diagnosis ; drug therapy ; etiology ; Female ; Hematopoietic Stem Cell Transplantation ; adverse effects ; Humans ; Male ; Middle Aged ; Polymerase Chain Reaction ; methods ; Retrospective Studies ; Transplantation, Homologous ; Young Adult

A fluorescent immunochromatographic test strip based on the quantum dots submicrobeads (QBs) was developed for quantitative detection of chloramphenicol (CAP). In this method, monoclonal antibody of CAP and OBs complex fluorescent probe was first prepared using 1-ethyl-3-( 3-dimethylaminopropyl ) carbodiimide / N-hydroxysuccinimide coupling approach, then complete antigen CAP-HS-BSA was synthesized and sprayed on nitrocellulose membrane as test line (T line). Similarly, goat anti-mouse antibody was sprayed as control line (C line). The time required for the analysis was 15 min, and the limit of detection (LOD) for CAP was 0. 1 μg / L, with a working range of 0. 1 - 100 μg / L. In spiked milk samples, the test strip demonstrated high recoveries in the range from 93. 3% to 97. 9% with relative standard deviations of less than 7% .