ObjectiveTo detect the mRNA expression levels of RORγt and Foxp3 genes in peripheral blood mononuclear cells(PBMCs) of children with atopic dermatitis(AD),and to explore their possible roles in the development of AD.MethodsSixty-three children with AD and 54 age-matched normal children were eligible for this study.Real-time fluorescence-based quantitative PCR was carried out to measure the mRNA expressions of RORγt and Foxp3 genes in PBMCs of these subjects.ResultsThe relative expression levels of Foxp3 and RORγt mRNA were significantly higher in children with AD compared with the normal controls (2.4060 ± 0.3355 vs.1.1852 ± 0.4189,t =17.50,P＜ 0.01; 6.9130 ± 0.2046 vs.2.7501 ± 0.2518,t =98.63,P ＜ 0.01 ).A statistical increase was observed in the level of RORγt mRNA in children with severe AD compared with children with mild and moderate AD(7.1203 ± 0.1056 vs.6.8046 ± 0.1731 and 6.8655 ± 0.3671,t =6.61,5.23,both P ＜ 0.01).No significant difference was noted in the expression of RORγt mRNA between children with mild and moderate AD,or in the Foxp3 mRNA expression among children with mild,moderate and severe AD.The severity of AD was positively correlated with the level of RORγt mRNA (r =0.62,P ＜ 0.01 ),but uncorrelated with the level of Foxp3 mRNA(r =0.04,P ＞ 0.05).ConclusionsThe high expressions of RORγt and Foxp3 genes may be involved in the pathogenesis of AD,and the severity of AD is positively correlated with the expression intensity of RORγt gene.

Objective To introduce a new septorhinoplastic method for twisted nose, humped nose and hawk nose as well as deviation of nasal septum, and to evaluate its operative method and effect. Methods The incision of cleft between lateral nasal cartilage and greater alar cartilage was employed in all patients. The method of unilateral mucoperiosteal flap osteotomy was used to correct nasal pyramid deformity, and swinging-door operation was performed to correct the deviation of nasal septum simultaneously. Results The operative results of 36 cases were effective and satisfactory. Postoperative follow-up ranged from one year to fifteen years. Based on the results of follow-up in the 32 cases, no recurrence and complications were found in all the cases. Conclusion This new method for septorhinoplasty avoids the detachment and move of nasal bone and nasal septum cartilage and injury of nasal mucoperiosteum, and maintains the blood supply of nasal bone and nasal septum cartilage. It corrects extero-nasal deformity as well as deviation of nasal septum. The operation is effective in nose aesthetic and ventilatory function.

Objective To investigate the clinical effects of montelukes in treatement of wheezing infants with viral infections. Methods 90 wheezing infants with viral infections were divided into oral Montelukas treatment( Group A) 50 cases and inhaled corticosteroid treatment (Group B) 40 cases, Group B were treated with conventional therapy plus glucocorticoids, Group A in addition to therapy with Montelukast 4mg/time at a draught quaque nocte for 10 days. The clinical effect 、improvement of symptoms 、 pulmonary function, blood gas analysis and the side effects were observed for the infants in both groups. Results The viral infections-positive with first break in 70 cases (77. 7% ) were significantly higher than that recurrent break in 20 cases (22.2% ) (x2 =6. 65, P ＜ 0.01) ; The scores of PaO2 after treatment were significantly higher than before treatment in two groups( t = 2. 542,2.533,all P ＜ 0. 05 ) ; A total effective rate was 88. 0% in group A which was significantly higher than that 72. 5% in group B( x2 = 3. 751 ,P ＜ 0.05) ;The adverse reactions incidence 4. 0% (2/50) in group A was significantly lower than the B group 10. 0% (4/40) in group B(x2 =3.857,P＜0.05). Conclusion Montelukast had definitely effect for infant wheezing with virus infection, and had less side effects.

Objective To study the effect of whole-process nursing intervention on comfort of patients undergoing radial artery puncture. Methods 100 patients with radial artery puncture from July, 2007 to June, 2008 were divided into the control group and the experimental group with 50 cases in each group accord-ing to time sequence. Routine whole nursing mode was used in the control group, the whole-process nursing intervention mode based upon routine nursing mode under the instruction of evidence-based method was used in the experimental group. Pain, psychological tensity, body numbness, success rate of radial artery puncture, patients' satisfaction degree were compared between the two groups. Results Every indexes of the experi-mental group were better than those of the control group. Conclusions The whole-process nursing interven-tion mode can promote the comfort degree of patients undergoing radial artery puncture.

Objective To study the impact of narrow band ultraviolet B (NB-UVB) irradiation on the expression of T helper 17 (Thl7) and CD4,CD25 and regulatory T (Treg) cells and their related cytokines transfor-ming growth factor beta 1 (TGF-β1) and interleukin-6 (IL-6) to further clarify how NB-UVB treatment helps patients with psoriasis vulgaris.Methods Ninety patients with active stage and stationary stage psoriasis vulgaris (45 cases each) were treated with NB-UVB for 8 weeks.Fifty healthy persons were used as normal controls.Peripheral blood levels of Th17 and Treg were measured by flow cytometry.An enzyme-linked immunosorbent assay (ELISA) was used to measure the serum levels of TGF-β1 and IL-6 before and after treatment.Clinical efficacy was evaluated in terms of the area of psoriasis and severity index (PASI) scores.Restlts Before treatment,the patients showed significantly higher levels of Thl7 cells in their peripheral blood than the controls.Their ratios of Th17 to Treg cells and their serum levels of IL-6 were also significantly higher.The percentage of Treg cells and the serum level of TGF-β1 were significantly lower in the patients.After the NB-UVB treatment,the Th17 cells,the ratio of Th17 to Treg cells and IL-6 had all decreased significantly.The percentage of Treg cells and TGF-β1 levels were significantly high-er compared with before phototherapy.The total effectiveness rate was 86.7％,and the average PASI scores had de-creased significantly.The PASI scores were positively correlated with the percentage of Th17,the Th17 to Treg ratio,and the serum level of IL-6,and negatively correlated with the percentage of Treg cells and TGF-β1.Conclusion The imbalance between Th17 and Treg cells and their cytokines may play an important role in the pathogenesis of pso-riasis.NB-UVB is able to significantly down-regulate the levels of Th17,IL-6 and the progression of Treg levels and TGF-β1 expression.It can regulate the balance between Thl7and Treg cells,which may be one of the mechanisms of NB-UVB treatment for psoriasis.The clinical data demonstrate that NB-UVB treatment is a safe and effective therapy for psoriasis vulgaris.

Objective To observe clinical efficacy of NB-UVB in treating psoriasis vulgaris and its effects on expression of serum interleukin-17 (IL-17),interleukin-23 (IL-23) and interleukin-22 (IL-22) in patients with psoriasis vulgaris,and to study the underlying mechanisms of NB-UVB.Methods Ninety patients were recruited and treated with NB-UVB therapy for 8 weeks.Before and after treatment,the serum level of IL-17,IL-23,IL-22,interleukin-l0(IL-10) and transforming growth factor(TGF-β) were tested by use of ELISA method,meanwhile Psoriasis Area and Severity Index (PASI) were used to evaluate clinical efficacy.Fifty healthy volunteers were selected as control group.Results Compared to healthy controls,the level of serum IL-17,IL-23 and IL-22 was significantly higher in patients with psoriasis vulgris (P ＜ 0.01),and IL-10,TGF-β shown lower expression in psoriasis patients (P ＜ 0.01).After 8 weeks of treatment with NB-UVB,serum levels of IL-17,IL-23 and IL-22 in psoriasis patients decreased significantly (P ＜ 0.01),while IL-10,TGF-βelevated significantly (P ＜ 0.01) in contrast.The Psoriasis Area and Severity Index (PASI) results indicated significantly clinical improvement after therapy,and the total effective rate was 87.78％.Conclusion NB-UVB could down-regulate serum IL-17,IL-23,IL-22 and up-regulate IL-10,TGF-β,which may help regulate imbalance of T lymphocytes cells of psoriasis patients.The clinical data demonstrate that the treatment of NB-UVB is a safe,effective method for psoriasis vulgaris.

Objective To analyze the suspicious results of serum HBV DNA by fluorescence quantitative PCR and develop appropriate countermeasures in order to improve the quality of detection of HBV DNA.Methods Blood samples of patients from the First Affiliated Hospital of Wenzhou Medical College from 2008 to 2011 were analyzed for HBV DNA by fluorescence quantitative PCR.1969 cases of suspicious results,judged by the rule of review the results of serum HBV DNA combined with the historical results,PCR amplification curve,HBV serum markers and clinical diagnosis,were analyzed and redetected by using of two different reagents,careHBV PCR Kit and careHBV PCR Kit V2,at the same time.The consistency and inconsistency ratio of the results were evaluated.Both the reasons of inconsistent and the undetected rates of careHBV PCR Kit were analyzed.The two reasons for the inconsistent results included the reagent related factors,e.g,showing no amplification curve caused by the false negative and abnormal low efficiency of amplification curve,and the non reagent related factors such as operating pollution and other sample factors.Results There were 115 154 blood samples were detected for HBV from 2008 to 2011 and 1969 samples (1.71％) with suspicious results were redetected.The consistency and inconsistency results were 1588 (80.65％) and 381 (19.35％),respectively.Every year from 2008 to 2011,the percentage of the inconsistent results caused by the reagent related factors were 18.87％,20.23％,51.33％ and 59.57％ respectively,which showed an increasing trend,and the percentage of inconsistent results caused by the nonreagent related factors were 81.13％,79.77％,48.67％ and 40.43％ respectively,which showed a declining trend year by year.The undetected rates of careHBV PCR Kit were 2.49％,4.08％,10.09％ and 14.47％ respectively,showing an increasing trend.Conclusions The redetection for the specimens with the suspicious results by using of different reagents can avoid the blind detection of HBV DNA and reduce the experimental error.All the clinical samples for quantitative HBV DNA including the mutations of HBV gene can be measured accurately and effectively,which is helpful to hepatitis B patients for antiviral therapy.

Objective To explore the value of procalcitonin (PCT) in the prediction of the prognosis and severity of bacterial infection in critically ill patients.Methods A total of 116 eligible patients with bacterial infection admitted in the intensive care unit were enrolled in this prospective study from February,2012 through November,2012.Within 24 hours after admission,the serum PCT was determined with immune-chromatography and acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) score of patients was calculated.Based on the 28-day clinical outcome of patients,the patients were divided into fatal group (n =36) and survival group (n =80).The differences in PCT and APACHE Ⅱ score between the two groups were compared with t test or rank-sum test.The correlation between PCT and APACHE Ⅱ score was determined with Spearman's correlation analysis.Both PCT and APACHE Ⅱ score were analyzed separately and jointly with area under receiver operator characteristic curve (ROC curve,AUC) to predict 28-day survival.Comparison of prediction performance for predicting 28-day survival of patients with bacterial infection between PCT and APACHE Ⅱ was made with U test.Results PCT concentration was significantly higher in fatal group than that in survival group [5.36 (2.07,25) vs.0.24 (1.00,2.14)] (Z =5.596,P ＜0.01).APACHE Ⅱ score within 24 hour after admission was significantly higher in fatal group than that in survival group (24.30 ± 6.71) vs.(16.03 ± 7.23),t =6.147,P ＜ 0.01.Positive correlation between PCT and APACHE Ⅱscore was found to be statistical significance (r =0.388,P＜ 0.01).When rates of 28-day survival in patients were predicted by using PCT and APACHE Ⅱ score,the areas of under curve were 0.804 and 0.792,respectively.AUC of PCT was tenuously larger than that of APACHE Ⅱ score (U =0.2073,P =0.802).Using PCT and APACHE Ⅱ score together to predict 28-day survival,AUC (0.817) was increased.The joint prediction performance was higher than that of either alone,increasing the sensitivity to 90.7％ and the specificity to 75.2％.Conclusions Serum PCT can reflect the severity of the illness and prognosis of infectious disease in the intensive care unit.It can serve as a sensitive marker of predicting 28-day survival.Combining PCT and APACHE Ⅱ score together can increase the prognostic value.

Objective:To determine the effect of Notch1 signaling pathway on the differentiation and function of Th17 cells in murine psoriasis model.Methods: BALB/c mice were randomly divided into psoriasis model group and control group.Murine psoriasis model was established by topical 5% imiquimod application in combination with intraperitoneal injection of α-2b interferon.The CD4+ T lymphocytes were isolated by magnetic activated cell sorter (MACS).Flow cytometric analysis (FCM) was performed to detect the percentage of Th17 cells.Real-time RT-PCR was employed to measure the mRNA levels of RORγt,IL-17A,Notch1 and Hes-1.The CD4+ T lymphocytes were then divided into γ-secretase inhibitor DAPT groups and control group,and the expression differences of Notch1 signaling molecule and its target gene Hes-1 mRNA levels,Th17 cell percentage,RORγt and IL-17A mRNA levels,and IL-17A concentrations in cell-free supernatant were detected.Results: The expression levels of Th17 cell percentage and RORγt,IL-17A,Notch1 and Hes-1 mRNA in CD4+ T lymphocytes of murine psoriasis model were significantly higher than control mouse[(2.97±0.86)% vs.(0.65±0.11)%,t=15.083;(5.75±0.61) vs.(1.57±0.43),t=21.630;(7.83±0.97) vs.(1.63±0.31),t=25.348;(7.10±1.37) vs.(1.47±0.34),t=17.386;(7.30±1.15) vs.(1.67±0.48),t=18.840,respectively,all P<0.01].Compared with control group,Th17 cell percentage,mRNA expression levels of Notch1,Hes-1,RORγt and IL-17A,and IL-17A concentrations in cell-free supernatant from cultured CD4+ T lymphocytes of murine psoriasis model were dramatically decreased in DAPT treated groups in a dose-dependent way (F=74.368,89.719,126.572,94.558,124.323 and 123.231 respectively,all P<0.01).Conclusion: Notch1 signaling pathway can regulate the differentiation and function of Th17 cells in murine psoriasis model,and may have potential value for the target immunotherapy of psoriasis.

Objective To explore the role of Toll like receptor 2 (TLR2) in the pathogenesis of acne vulgaris. Methods Venous blood was collected from 30 normal human controls and 90 patients with acne vulgaris. The patients were equally divided into three groups, i.e., mild, moderate and severe groups, according to disease severity. Flow cytometry was performed to detect the expression of TLR2 in PBMCs, and double antibody sandwich ELISA to measure the levels of serum interleukin-8 (IL-8) and tumor necrosis factor-α (TNF-α). Results A significant increase was observed in the expression of TLR2 in PBMCs, serum level of IL-8and TNF-α in the three groups of patients with acne vulgaris compared with the normal human controls (all P ＜0.01). The expression of TLR2 was positively correlated with the expression of IL-8 (r=0.382, 0.517,0.436,respectively, all P＜0.05) and TNF-α(r=0.641, 0.725, 0.593, respectively, all P＜0.05) in the patients with mild, moderate and severe acne vulgaris, respectively, and with the severity of acne vulgaris (r = 0.406,P＜0.01 ). Conclusion The expression level of TLR2 and related cytokines seems closely correlated with the severity of acne vulgaris.