Objective: To observe the antiasthmaf ic, anti-inflammatory and immunological effects of Gubenchuansoukang Granule. Methods: The rats asthma model was established by ovalbumin sensitization. The model was identified by the study of the asthma incubation period and the total white cells counts in bronchoalveolar lavage fluid (BALF). The anti-inflammatory effect was observed by the acute inflammation and chronic inflammation induced xylene and agar. To observe the effect of Gubenchuansoukang Granule on humoral immunity and cellular immunity induced by chicken red blood cell and 2, 4-dinitrochlorobenzene. Results: Gubenchuansoukang Granule could lengthen the asthma incubation period, reduce the quantity of total white cells in BALF, against the acute inflammation and chronic inflammation, and restrain the humoral immunity and cellular immunity. Conclusion: Gubenchuansoukang Granule had antiasthmafic, anti-inflammatory and immunosuppression effects.

OBJECTIVETo observe the treatment effect and toxicity of nonmyeloablative allogeneic stem cell transplantation (NST) for hematologic diseases.

METHODSA total of 243 hematologic diseases patients received HLA-identical NST were enrolled in this study from 9 transplant centers of NST Cooperative Group in China. Nonmyeloablative conditioning regimen was based on fludarabine (Flud), rabbit anti-human thymocyte globulin (ATG), cyclophosphamide (CTX) (FAC), and plus cytarabine or busulfan (BU) etc. Graft-versus-host disease (GVHD) prophylaxis included cyclosporin A (CsA) and mycophenolate mofetil (MMF).

RESULTSAmong the 243 patients, 219 (90.1%) achieved full donor chimerism (FDC), 2(0.8%) engraftment failure. 78 (32.1%) had mixture chimerism (MC) at 4 weeks after NST, out of which 56 switched to FDC, 16 remained MC and 6 (2.5%) developed graft rejection. The incidence of acute GVHD was 34.2%, including 6.6% of grade III-IV acute GVHD. Chronic GVHD developed in 78 (32.1%) patients. The follow-up durations were 3 - 99 months, 162 (66.7%) were still alive and the overall survival rates were 76.5%, 73.9%, 70.7%, and 27.8% for MDS/SAA, chronic myeloid leukemia, acute leukemia at first remission, and refractory or relapsed leukemia, respectively.

CONCLUSIONSThe nonmyeloablative allogeneic stem cell transplantation based on FAC conditioning results in sustained engraftment and mild aGVHD, providing a new feasible curative therapy for hematology diseases.

Adolescent ; Adult ; Child ; China ; Female ; Graft vs Host Disease ; prevention & control ; Hematologic Diseases ; surgery ; Hematopoietic Stem Cell Transplantation ; methods ; Humans ; Male ; Middle Aged ; Transplantation Conditioning ; methods ; Transplantation, Homologous ; Treatment Outcome ; Young Adult

OBJECTIVETo express and purify the protein coded by the TRAF-type zinc finger domain of myasthenia gravis (MG)-related gene P9 (P9-ZFD) and to prepare P9-ZFD antiserum for detecting expression and subcellular distribution of P9-ZFD protein in the skeletal muscles of patient with MG.

METHODSThe cDNA encoding P9-ZFD was amplified by RT-PCR. The cloned P9-ZFD cDNA was ligated into pET24a, and the P9-ZFD recombinant protein was induced via E. coli. BL21 (DE3) and purified by histidine affinity chromatography. P9-ZFD antiserum was prepared and its titer and specificity were determined by ELISA and Western blot. Expression and subcellular distribution of P9-ZFD protein in the skeletal muscles of MG and control were studied.

RESULTSThe molecular weight of purified P9-ZFD protein was about 30 kD. Its purity was more than 95%. Antiserum specific for P9-ZFD was excellent. P9-ZFD protein is fully confined to the cytoplasm membrane of skeletal muscle cell of MG, obvious immunostaining was absent in the A, I, and Z bands of cytoplasm and no immunoreactivity was observed in the skeletal muscle cell of control.

CONCLUSIONP9-ZFD protein is expressed as a cytoplasm membrane-bound protein and has obvious distribution difference in the skeletal muscle cells of patient with MG and normal control.

Adult ; Cell Membrane ; metabolism ; Escherichia coli ; metabolism ; Female ; Humans ; Muscle Proteins ; biosynthesis ; genetics ; Muscle, Skeletal ; metabolism ; pathology ; Myasthenia Gravis ; metabolism ; Peptide Fragments ; biosynthesis ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Transfection ; Zinc Fingers

Adult ; Aged ; Hemospermia ; diagnostic imaging ; therapy ; Humans ; Male ; Middle Aged ; Punctures ; Rectum ; diagnostic imaging ; Seminal Vesicles ; Therapeutic Irrigation ; Ultrasonography

OBJECTIVETo evaluate the anticancer effects of exogenous human WT-PTEN overexpression on bladder transitional carcinoma cell line EJ.

METHODSThe plasmid containing WT-PTEN or mutant PTEN was separately transfected into bladder transitional carcinoma cell line EJ, and the protein expression of PTEN in the EJ cells was detected by Western blot. Cell morphological changes were observed under the inverted microscope and transmission electron microscope. MTT test was used to assess the effect of PTEN on proliferation and anticancer effects for mitomycin and theraubicin. The change of bcl-2 expression in the cells was measured by Western blot. The empty plasmid was used as control.

RESULTSWestern blot analysis showed that EJ cells expressed high level of PTEN protein after transfection with WT-PTEN or mutant PTEN plasmid. Abnormal morphological changes of the cells were observed in WT-PTEN transfected groups. The growth of EJ cells treated with WT-PTEN was significantly inhibited by 40.1% and anticancer effects were enhanced by mitomycin and theraubicin, but the cells transfected with mutant PTEN plasmid did not show such similar biological behavior.

CONCLUSIONWT-PTEN gene transfection can suppress the in vitro growth and induce apoptosis of bladder transitional carcinoma cell line EJ cells. Mutant PTEN does not show similar biological behavior. Overexpression of WT-PTEN inhibits cancer cell proliferation by down-regulating bcl-2 expression in the cells.

Antibiotics, Antineoplastic ; pharmacology ; Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Blotting, Western ; Carcinoma, Transitional Cell ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Doxorubicin ; analogs & derivatives ; pharmacology ; Green Fluorescent Proteins ; genetics ; metabolism ; Humans ; Microscopy, Electron, Transmission ; Mitomycin ; pharmacology ; Mutation ; PTEN Phosphohydrolase ; genetics ; metabolism ; physiology ; Plasmids ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Recombinant Fusion Proteins ; genetics ; metabolism ; physiology ; Transfection ; Urinary Bladder Neoplasms ; genetics ; metabolism ; pathology

OBJECTIVETo evaluate the various operative details of strictures of the posterior urethra that are essential for a successful result.

METHODSThe clinical data of 191 patients with posterior urethral strictures or distraction defects from January 1990 to January 2006 were analyzed retrospectively. All patients underwent a retrograde and voiding urethrogram, 62 patients had urethral ultrasonography, 48 patients had urethroscopy, 3 patients had MRI. Repair was performed with a simple anastomosis after urethral mobilization in 66 patients, separation of the corporeal bodies in 48 patients, separation of the corporeal bodies and inferior pubectomy in 30 patients, transpubic anastomosis in 18 patients, pull-through operation in 3 patients, and optical urethrotomy in 26 patients. Followup ranged from 6 to 48 months.

RESULTSThe mean stricture length was 3.6 cm (range from 1.5 to 8.0 cm). Posterior urethral strictures is in 31 (16%), posterior distraction defects is in 160 (84%), of which the length of the distraction defects < 3 cm is in 102 (53.4%), and the the length of the distraction defects > 3 cm is in 58 (30.6%). The overall successful results (Qmax > 15 ml/s) after operation was 84.3%. Optical urethrotomy was 69%, the successful results with anastomotic urethroplasty were 97% with a simple anastomosis; 79% with separation of the corporeal bodies; 80% with separation of the corporeal bodies and inferior pubectomy; 83% with transpubic anastomosis; and 67% with pull-through operation.

CONCLUSIONSThe anastomotic urethroplasty is better than the optical urethrotomy, the length of the strictures or distraction defect which is lower than 3 cm is much more successfully corrected.

Adolescent ; Adult ; Aged ; Child ; Female ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Treatment Outcome ; Urethral Stricture ; surgery ; Urologic Surgical Procedures, Male ; methods

OBJECTIVETo study the expression of Toll-like receptor 4 (TLR4) in renal tubular epithelial cells exposed to high glucose and the effect of spironolactone on the TLR4 expression.

METHODSIn vitro renal tubular epithelial cells (NRK-52E) were randomly exposed to DMEM culture solution with low glucose (5 mmol /L), high glucose (25 mmol/L) or 10(-7) mol/L spironolactone plus 25 mmol/L glucose. Immunohistochemistry, RT-PCR and Western blot were used to determine TLR4 protein and mRNA expression. The levels of IL-6 and TNF-alpha in the cell culture supernatant were determined using ELISA.

RESULTSThe expression of TLR4 mRNA in the high glucose group began to increase 6 hrs and remained at a higher level up to 24 hrs after exposure as compared with the low glucose group. The TLR4 mRNA expression in the spironolactone treatment group was significantly lower than that in the high glucose group, although it was higher than that in the low glucose group between 6 and 24 hrs after exposure. TLR4 protein expression increased significantly in the high glucose group 24 and 48 hrs after exposure compared with that in the low glucose group. The TLR4 protein expression in the spironolactone treatment group was lower than that in the high glucose group, but higher than that in the low glucose group. IL-6 and TNF-alpha expression in the supernatant from the NRK-52E cells in the high glucose groups increased significantly as compared with the low glucose group. The spironolactone treatment group had significantly reduced IL-6 and TNF-alpha expression compared with the high glucose group.

CONCLUSIONSHigh glucose triggers an increase in the expression of TLR4 and inflammatory factors in NRK-52E cells. TLR4 may participate in the progress of diabetic nephropathy. Spironolactone can reduce expression of TLR4 and inflammatory factors, which might be attributed to one of the mechanisms of protection by spironolactone against diabetic nephropathy.

Cells, Cultured ; Diabetic Nephropathies ; etiology ; Epithelial Cells ; metabolism ; Humans ; Hyperglycemia ; metabolism ; Immunohistochemistry ; Interleukin-6 ; analysis ; Kidney Tubules ; metabolism ; RNA, Messenger ; analysis ; Spironolactone ; pharmacology ; Toll-Like Receptor 4 ; analysis ; genetics ; physiology ; Tumor Necrosis Factor-alpha ; analysis

OBJECTIVETo evaluate the selection of different procedures and the feasibility for the treatment of long segment urethral stricture.

METHODSSeventy-six patients with complex urethral stricture greater than 8 cm long underwent different procedures of urethroplasty. Of them various mucosa grafts urethral reconstruction were adopted in 42 cases (colonic mucosal graft, n = 26; buccal mucosal graft, n = 10; bladder mucosal graft, n = 6); One-stage pedicle flaps urethroplasty in 20; two-stage urethroplasty of Johanson procedure in 12; and penile urethra-prostatic urethra anastomosis, three-stage urethroplasty in 2.

RESULTSIn early followed up (within 6 months postoperatively), 67 patients (88%) voided well and complications developed in 10. Among the 70 patients who lasted more than 1 year after operation, 51 cases were followed up. Forty-four patients voided well, and complications developed in 8. Of the 8 cases urethral restructure developed in 2 (18%) for pedicle flaps urethroplasty, 2 for colonic mucosal urethroplasty (9%), 1 for buccal mucosal graft (1/7), 1 for bladder mucosal graft (1/3); penile chordee in 2 (2/5), and one of them was accompanied by hair bearing neourethra for two-stage urethroplasty of Johanson procedure.

CONCLUSIONSColonic mucosal and buccal mucosal grafts urethroplasty are feasible procedures for the treatment of long segment urethral stricture, and Colonic mucosal graft urethroplasty may be considered when more conventional procedures fail or complicated urethral strictures greater than 10 cm long.

Adolescent ; Adult ; Aged ; Follow-Up Studies ; Humans ; Intestinal Mucosa ; surgery ; Male ; Middle Aged ; Mouth Mucosa ; surgery ; Surgically-Created Structures ; Treatment Outcome ; Urethral Stricture ; pathology ; surgery ; Urologic Surgical Procedures, Male ; methods

OBJECTIVETo establish an assessment index system to objectively evaluate the implementation of health promotion in enterprises.

METHODSMulti-methods, which include the reviewing references, the summarizing results of preliminary studies, the interview with experts and employers, were used in developing attentative index framework and working out the consultative questionnaires. Then the improved Delphi Method was adopted in collecting ideas from 20 experts in China, and they scored the importance of every single index by two rounds of consultation. On the basis of these data, the indicators were added , deleted or modified according to concentration and distraction levels of expert ideas. The method of the percentile weighted coefficient was conducted to decide the weighing of assessment index.

RESULTSThe responding rates to the two rounds of questionnaires were 90.91% and 100.00% respectively. The overall specialist authoritative coefficient was 0.69, the coordination coefficient of all indicators was 0.623 (P <0.01),and the coordination coefficient of the four kinds of first-ranking indicators was 0.313, 0.625, 0.390, 0.563, respectively (P<0.01).Eventually, an assessment index system consisting of 4 first-ranking indicators and 10 sub-indicators which were further divided into 52 grade-three indicators was set up.

CONCLUSIONThe assessment index system possesses good content validity, and the liability and typicality were recognized by experts. However, it should be applied and validated in practice.

Adult ; China ; Delphi Technique ; Health Promotion ; Humans ; Middle Aged ; Occupational Health Services

BACKGROUNDThe interaction between activated microglia and T lymphocytes can yield abundant pro-inflammatory cytokines. Our previous study proved that thymus immune tolerance could alleviate the inflammatory response. This study aimed to investigate whether intrathymic injection of myelin basic protein (MBP) in mice could suppress the inflammatory response after co-culture of T lymphocytes and BV-2 microglia cells.

METHODSTotally, 72 male C57BL/6 mice were randomly assigned to three groups (n = 24 in each): Group A: intrathymic injection of 100 μl MBP (1 mg/ml); Group B: intrathymic injection of 100 μl phosphate-buffered saline (PBS); and Group C: sham operation group. Every eight mice in each group were sacrificed to obtain the spleen at postoperative days 3, 7, and 14, respectively. T lymphocytes those were extracted and purified from the spleens were then co-cultured with activated BV-2 microglia cells at a proportion of 1:2 in the medium containing MBP for 3 days. After identified the T lymphocytes by CD3, surface antigens of T lymphocytes (CD4, CD8, CD152, and CD154) and BV-2 microglia cells (CD45 and CD54) were detected by flow cytometry. The expressions of pro-inflammatory factors of BV-2 microglia cells (interleukin [IL]-1β, tumor necrosis factor-α [TNF-α], and inducible nitric oxide synthase [iNOS]) were detected by quantitative real-time polymerase chain reaction (PCR). One-way analysis of variance (ANOVA) and the least significant difference test were used for data analysis.

RESULTSThe levels of CD152 in Group A showed an upward trend from the 3rd to 7th day, with a downward trend from the 7th to 14th day (20.12 ± 0.71%, 30.71 ± 1.14%, 13.50 ± 0.71% at postoperative days 3, 7, and 14, respectively, P < 0.05). The levels of CD154 in Group A showed a downward trend from the 3rd to 7th day, with an upward trend from the 7th to 14th day (10.00 ± 0.23%, 5.28 ± 0.69%, 14.67 ± 2.71% at postoperative days 3, 7, and 14, respectively, P < 0.05). The ratio of CD4+/CD8 + T in Group A showed a downward trend from the 3rd to 7th day, with the minimum at postoperative day 7, then an upward trend from the 7th to 14th day (P < 0.05). Meanwhile, the levels of CD45 and CD54 in Group A were found as the same trend as the ratio of CD4+/CD8 + T (CD45: 83.39 ± 2.56%, 82.74 ± 2.09%, 87.56 ± 2.11%; CD54: 3.80 ± 0.24%, 0.94 ± 0.40%, 3.41 ± 0.33% at postoperative days 3, 7, and 14, respectively, P < 0.05). The expressions of TNF-α, IL-1β, and iNOS in Group A were significantly lower than those in Groups B and C, and the values at postoperative day 7 were the lowest compared with those at postoperative days 3 and 14 (P < 0.05). No significant difference was found between Groups B and C.

CONCLUSIONSIntrathymic injection of MBP could suppress the immune reaction that might reduce the secondary immune injury of brain tissue induced by an inflammatory response.

Animals ; Anti-Inflammatory Agents ; pharmacology ; Antigens, Surface ; analysis ; Brain Injuries, Traumatic ; drug therapy ; CD4-CD8 Ratio ; Coculture Techniques ; Male ; Mice ; Mice, Inbred C57BL ; Microglia ; immunology ; Myelin Basic Protein ; administration & dosage ; pharmacology ; T-Lymphocytes ; immunology