gic rhinitis animal model Al(OH)3 colloid gel of different concentration (0.5-5 mg) may cause side effects such as foreign body granuloma.

BACKGROUNDCurrently anti-inflammatory therapy with steroids for allergic rhinitis need long-term repeated administration, although it is effective. Gene therapy is being suggested to substitute it. The aim of this study was to investigate nonviral vector mediated exogenous gene expression in COS-7 cells in vitro and the effect of intranasal mouse interleukin (mIL)-12 transgene expression on allergen induced eosinophil infiltration of nasal mucosa in a murine model of allergic rhinitis.

METHODSIn vitro COS-7 cells were infected with Epstein-Barr virus (EBV)/lipoplex. The expression of IL-12 p70 in cell culture supernatant was examined by enzyme-linked immunosorbent assay (ELISA). In mice with ovalbumin (OVA) induced allergic rhinitis, EBV/lipoplex was administered by nasal drops before OVA challenge once a day from day 1 to day 10. The expression of IL-12 mRNA and protein, the change of eosinophil count in nasal mucosa and serum total IgE were measured 24 hours after the last challenge.

RESULTSEBV/lipoplex could effectively transfect COS-7 cells. The expression of IL-12 p70 in cell culture supernatant was significantly more than in blank control. IL-12 via EBV plasmid vector transduction could be overexpressed in vivo. In pGEG.mIL-12 treated models, the nasal mucosa revealed a high level of widespread mIL-12 transduction by immunohistochemistry and in situ hybridization. Histological evaluation revealed marked suppression of eosinophil infiltration in nasal mucosa. The eosinophil count in allergic rhinitis group [(26.5 +/- 9.8)/high-power field (HPF)] was significantly increased over control group [(0.40 +/- 0.52)/HPF] (F = 56.94, P < 0.01), while the count in IL-12 gene therapy group [(4.60 +/- 2.63)/HPF] was significantly less than that of allergic group (F = 56.9, P < 0.01). Serum total IgE between in gene therapy mice [(88.83 +/- 6.71) ng/ml] and allergic rhinitis mice [(103.1 +/- 5.7) ng/ml] showed a significant difference (F = 1216, P < 0.05).

CONCLUSIONSNonviral EBV plasmid vector, pGEG.mIL-12 was able to overexpress exogenous gene both in vitro and in murine nasal mucosa in vivo. IL-12 overexpression via EBV/lipoplex could stem allergen induced eosinophil infiltration in nasal mucosa in murine models of allergic rhinitis, which may suggest a new cytokine immunogenetic therapy for allergic rhinitis.

Administration, Intranasal ; Animals ; COS Cells ; Cercopithecus aethiops ; Eosinophilia ; therapy ; Genetic Therapy ; Herpesvirus 4, Human ; genetics ; Immunoglobulin E ; blood ; Interleukin-12 ; genetics ; Lipids ; administration & dosage ; Male ; Mice ; Mice, Inbred BALB C ; Nasal Mucosa ; metabolism ; Rhinitis, Allergic, Perennial ; therapy ; Rhinitis, Allergic, Seasonal ; therapy

Objective To explore the influence of sputum aspiration appliance on respiratory tract infection of patients with tracheotomy. Methods 34 patients with tracheotomy from June 2008 to May 2009 (the first stage) and 30 patients with tracheotomy from June 2009 to May 2010 (the second stage) were analyzed retrospectively. The difference of the rates of species of infection pathogens and pneumonia complications in two stages were compared. Results In two stages, there was a significant difference of multiple antibiotic resistant strains respectively in the first week (acinetobacter baumannii, S. epidermidis, Pseudomonas aeruginosa) and in the second week (acinetobacter baumannii, S. epidermidis, Pseudomonas aeruginosa, Klebsiella pneumoniae,Staphylococcus aureus) (P < 0. 05 ). The rates of pneumonia complications in the second stage is lower than the first stage, the difference was significant (P < 0. 05 ). Conclusions It is an important component element tosterilize and isolate the sputum aspirators to decrease the incidence of Pulmonary multiple antibiotic resistant strains infection of patients with tracheotomy.

OBJECTIVETo investigate whether the local application of IL-12 gene with EBV-plasmid vector to nasal mucosa could prevent allergic inflammation in murine allergic rhinitis model.

METHODSThirty-six BALB/C mice were randomly divided into allergic rhinitis group gene therapy group and control group. In mice with OVA-induced allergic rhinitis, the EBV/lipoplex (a novel cationic lipid combined with EBV-plasmid vector, pGEG. mIL-12) was locally administered into nasal mucosa before OVA challenge. The expression of IL-12 mRNA and protein, the change of eosinophilia and mast cell, and Th2 cytokine production in the nasal mucosa were measured.

RESULTSThe amounts of IL-12 mRNA positive cells and IL-12 positive cells in nasal mucosa of gene therapy group were significantly higher than that of allergic rhinitis group (P < 0.01 and P < 0.05). The amount of eosinophils, mast cells, and the level of IL-5 expression in nasal mucosa in allergic rhinitis group were significantly higher than those in gene therapy group and control group (P < 0.01). The level of total IgE of peripheral blood in allergic rhinitis group was significantly higher than that in gene therapy group and control group (F = 1216.21, P < 0.01).

CONCLUSIONSThese findings indicated that IL-12 mRNA and protein were expressed effectively after the local administration of pGEG. mIL-12 in the nasal mucosa. The local application of pGEG. mIL-12 is effective in modulating nasal allergic response and may be a convenient method for future approach to allergic rhinitis.

Animals ; Genetic Therapy ; Genetic Vectors ; Interleukin-12 ; genetics ; therapeutic use ; Male ; Mice ; Mice, Inbred BALB C ; Nasal Mucosa ; metabolism ; Rhinitis, Allergic, Perennial ; therapy

OBJECTIVETo analyze the impact of olfactory nerve transection on the apoptosis of mice olfactory receptor neurons (ORN), and discuss the reliability of this experimental model.

METHODSAfter olfactory nerve transection of mice, anterograde horseradish peroxidase (HRP) tracing was performed to confirm the completion of nerve transection. On 8 h, 2 d, 3 d and 5 d after surgery, TdT mediated deoxyuridine triphosphate-biotin nick end labelling (TUNEL) was used to observe the apoptosis of ORN, while relative semi-quantitative RT-PCR and immunohistochemistry were used to reflect the expression of olfactory marker protein (OMP, special marker of mature ORN) in olfactory epithelium.

RESULTSNo HRP label was observed in olfactory bulb after olfactory nerve transaction. Both TUNEL-positive and OMP-positive cells were ORN. After the surgery, TUNEL-positive cells increased remarkably and peaked on 2 d after the surgery. Meanwhile OMP mRNA in olfactory epithelium began to decrease markedly till 5 d after the surgery, and the olfactory epithelium got thinner accordingly.

CONCLUSIONSThis experimental model can be used reliably to sever mice olfactory nerve and manipulate simultaneous apoptosis of mice ORN.

Animals ; Apoptosis ; Denervation ; Mice ; Mice, Inbred C57BL ; Olfactory Nerve ; cytology ; metabolism ; surgery ; Olfactory Receptor Neurons ; metabolism ; pathology

OBJECTIVETo find out the occurrence of cesarean section (CS) and to probe the factors associated with CS.

METHODSWomen with CS as "case group" and women without CS as "control group" were chosen in a case-control study.

RESULTSAmong 14 071 childbirth women, 6 421 had CS (case group) with the occurrence rate of 45.6% and 7 650 (54.4%) had normal delivery (control group). In comparison with the control group, the CS group had following several higher rates [with significant differences between case group and control group (P < 0.01)]: well-educated (78.9% vs 69.5%), white collar jobs (38.0% vs 32.3%), urban residents (79.1% vs 70.6%), high monthly income (>/= 500 Yuan) (81.0% vs 70.6%), of older age (>/= 25 years) (73.3% vs 63.0%), heavier baby weight (> 4 000 gram) (8.3% vs 2.9%), male babies (53.9% vs 51.4%), BMI of mother (> 24) (8.8% vs 4.8%), cephalopelvic disproportion (21.1% vs 0.9%), intrauterine asphysia (20.3% vs 6.7%), abnormality of force of labor (4.2% vs 2.7%), prolonged labor (2.9% vs 1.0%) and placenta previa (1.4% vs 0.4%). Our study also indicated that the higher the educational level was, the higher the rate of CS appeared; and the older the pregnant women was, the higher the rate of CS was. In CS group, over 70% primipara were over 24 years, and over 20% primipara had cephalopelvic disproportion and over 20% had intrauterine asphysia in CS group.

CONCLUSIONSAt present, the occurrence rate of cesarean section was rather high (45.6%) in China. The high rate of CS was more likely to associate not only with abnormal physiological/medical factors (eg. cephalopelvic disproportion, intrauterine asphysia, abnormality of force of labor, and prolonged labour, etc.), but also with some demographic factors as education, occupation, income and age, etc. It is necessary to take measures to reduce the unnecessary CS in China.

Adult ; Cesarean Section ; statistics & numerical data ; China ; Female ; Humans ; Logistic Models ; Pregnancy

OBJECTIVETo establish a method for inducing apoptosis of rhesus peripheral blood lymphocytes (PBLs).

METHODSRhesus PBLs were irradiated with X-ray, (60)Co gamma-rays and ultraviolet (UVC254 nm), respectively, and the cell apoptosis was evaluated with flow cytometry using annexin-V staining and propidium iodide staining.

RESULTSX-ray and (60)Co gamma-ray irradiation induced only low apoptotic rates of the PBLs, and UVC resulted in the highest apoptotic rate of about 60%. UVC irradiation of the PBLs in RPMI supplemented with 10% heat-inactivated fetal calf serum for 60 min at a distance of 20 cm led to an early apoptotic rate of 58.85% and necrotic rate of 11.5%. The apoptotic rate of PBLs increased in a dose- and time-dependent fashion.

CONCLUSIONFor inducing apoptosis of the rhesus PBLs, UVC can be more effective than X-ray and (60)Co gamma-ray. The highest apoptotic rate can be achieved when the rhesus PBLs in RPMI supplemented with 10% heat-inactivated fetal calf serum are exposed to UVC for 60 min at the distance of 20 cm.

Animals ; Apoptosis ; radiation effects ; Cells, Cultured ; Dose-Response Relationship, Radiation ; Flow Cytometry ; Gamma Rays ; Leukocytes, Mononuclear ; cytology ; radiation effects ; Lymphocytes ; cytology ; radiation effects ; Macaca mulatta ; Male ; Time Factors ; Ultraviolet Rays ; X-Rays

OBJECTIVETo investigate the clinical features, treatment modalities and the prognosis of nasal type NK/T cell lymphoma.

METHODSThe data of 39 such patients treated from June 2000 to December 2003 were retrospectively reviewed. Twenty three patients were treated by combined chemoradiotherapy, basing on anthracycline-containing CHOP or similar regimens (median 5 cycles). Eleven patients by chemotherapy alone, 2 by radiotherapy alone and 2 aged patients by palliative chemotherapy or radiotherapy. Radiotherapy was given by high energy photon ray combined with electron beam with a median curative dose of 56 Gy in conventional fractionation. Bivariate correlations and univariate prognostic factors were analyzed.

RESULTSMedian follow-up time for the 21 patients who were still alive was 22.5 months. The overall remission rate (RR) after initial treatment was 66.7% (21 CR, 3 PR). Chemotherapy alone got a CR rate of only 37.5%. The overall local control rate was 59.4%. Local relapse rate after curative radiotherapy was 25.0%. Radiotherapy was positively correlated with local control (P = 0.000) and time to disease progression (TTP, P = 0.002). Skin and intestine were among the extranodal relapse sites. Fifteen patients had highly aggressive tumors with a median survival time of only 5 months. Univariate analysis showed that significant favorable survival prognostic factors were: radiotherapy (P = 0.001); lower risk International Prognostic Index (IPI, P = 0.001); complete remission after primary treatment (P = 0.000); pre-diagnostic history > 2 months (P = 0.024); and free of skin involvement (P = 0.034).

CONCLUSIONMost of nasal type NK/T cell lymphoma are in early stage when diagnosed. Radiotherapy remains to be the mainstay of treatment. Combined chemoradiotherapy needs further improvement for the progressive disease type. Some patients may have highly aggressive tumors with poor prognosis. Optimal prognostic factors and individualized treatment regimens need to be investigated.

Adult ; Aged ; Aged, 80 and over ; Combined Modality Therapy ; Female ; Follow-Up Studies ; Humans ; Killer Cells, Natural ; Lymphoma, T-Cell ; pathology ; therapy ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; Nose Neoplasms ; pathology ; therapy ; Prognosis ; Remission Induction ; Retrospective Studies

OBJECTIVETo explore the operational mechanisms and potential approach to inducing transplantation immune tolerance of FTY720.

METHODSMouse splenocytes were incubated with FTY720, then the DNA was extracted and analyzed using gel electrophoresis. Hearts of inbred BALB/c (H-2(d)) mice were transplanted heterotopically in C57BL/6 (H-2b) mice. Recipients were randomly divided into six groups. Group-1 (n = 6) was the nil-treated control. Groups-2, 3 and 4 were given FTY720 at the dose of 3 mg.kg(-1) by oral gavage once a day with different time courses. Group-2 (n = 14) were administrated from 3 days before transplantation (day-3) to the 11th day after the transplantation (day 11); Group-3 (n = 6) from day 0 to day 14; Group-4 (n = 6) from day-3 to day 0. Group-5 (n = 5) and 6 (n = 5) were treated with Cyclosporine A (10 mg.kg(-1)) and 40-O-(2-hydroxyethyl)-rapamycin (RAD) (3 mg.kg(-1)) respectively by daily gavage from day 3 to day 11. The long survivors (> 100 d) in Group-2 were detected with their IL-4 and IFN-gamma levels and their tolerant state was challenged with second graft: the donor type skin.

RESULTSApoptosis changes of the mouse splenocytes incubated with FTY720 was showed by typical DNA ladders. The median survival time (MST) of Group-1 was 8 d. MST of Group-2 was 55 d and grafts in six mice survived more than 100 d. MST of Group-3 was 16.5 d. Group-4 has a MST of 14 d with one case exceeded 100 d. MST of Group-5 and 6 were 10 d and 13 d respectively. Long survivors of Group-2 can accept donor-type skin graft and the level of IL-4 in their serum is up-regulated while IFN-gamma remained stable.

CONCLUSIONSPretreatment of FTY720 bring about effect on the early events of transplantation immune responses. This effect might be mediated by apoptosis induction in lymphocytes using this drug. We originally designed the regime of FTY720 monotherapy, which started pre-operationally and maintained for a short period of time, and induced stable tolerance the allo-graft in mouse.

Adjuvants, Immunologic ; pharmacology ; Animals ; Apoptosis ; Fingolimod Hydrochloride ; Heart Transplantation ; immunology ; Immune Tolerance ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Myocardium ; pathology ; Propylene Glycols ; pharmacology ; Sphingosine ; analogs & derivatives ; Transplantation Immunology

Objective To establish the finite element model of spinal canal reconstruction and internal fixation,analysis influence of spinal canal reconstruction and internal fixation on spinal stability,and verify the effectiveness and reliability of spinal canal reconstruction and internal fixation in spinal canal surgery.Methods A 30-year-old male healthy volunteer with a height of 172 cm and weight of 75 kg was selected and his lumbar CT data were collected to establish a finite element model of normal lumbar Lo3-L,and the results were compared with in vitro solid results and published finite element analysis results to verify the validity of the model.They were divided into normal group,laminectomy group and spinal canal reconstruction group according to different treatment methods.Under the same boundary fixation and physiological load conditions,six kinds of ac-tivities were performed,including forward bending,backward extension,left bending,right bending,left rotation and right rota-tion,and the changes of range of motion(ROM)of L3-L4,L4-L5 segments and overall maximum ROM of L3-L5 were analyzed under the six conditions.Results The ROM displacement range of each segment of the constructed L3-L5 finite element model was consistent with the in vitro solid results and previous literature data,which confirms the validity of the model.In L3-L4,ROM of spinal canal reconstruction group was slightly increased than that of normal group during posterior extension(＞5％dif-ference),and ROM of other conditions was similar to that of normal group(＜5％difference).ROM in laminectomy group was significantly increase than that in normal group and spinal canal reconstruction group under the condition of flexion,extension,left and right rotation.In L4-L5,ROM in spinal canal reconstruction group was similar to that in normal group(＜5％differ-ence),while ROM in laminectomy group was significantly higher than that in normal group and spinal canal reconstruction group(＞5％difference).In the overall maximum ROM of L3-L5,spinal canal reconstruction group was only slightly higher than normal group under the condition of posterior extension(＞5％difference),while laminectomy was significantly higher than normal group and spinal canal reconstruction group under the condition of anterior flexion,posterior extension,left and right rotation(＞5％difference).The changes of each segment ROM and overall ROM of L3-L5 showed laminectomy group＞spinal canal reconstruction group＞normal group.Conclusion Laminectomy could seriously affect biomechanical stability of the spine,but application of spinal canal reconstruction and internal fixation could effectively reduce ROM displacement of the responsi-ble segment of spine and maintain its biomechanical stability.