Objective To investigate the effect of δ-aminolevulinic acid(ALA)-induced photodynamic reaction on biological characteristics of skin fibroblasts.Methods Human skin fihroblasts were isolated from the foreskin of children and cultured in the presence of 0.5 to 5.0 mmol/L ALA;three hours later,they were irradiated with 632.8 am laser followed by additional 12-hour culture.Then,MTI"assay and flow cytometry were used to measure the proliferation and apoptosis of cells.The levels of matrix metallopmteinase(MMP)1,-2 and-3 and hydroxyproline in the culture supematant of irradiated cells were determined by avidinbiotin complex-based ELISA and alkaline hydrolysis-based method,respectively.Results With the rise of ALA concentration from 0.5 to 5.0 mmol/L.the proliferation level of fibroblasts expressed as the absor-bance at 570 nm dropped from 0.45±0.05 to 0.32±0.04.and cell death rate increased from 6.4%±2.0% to 29.6%±2.2%.Meanwhile,the contents of MMP-1,-2 and -3 increased at the early stage,but decreased at the late stage,whereas the hydroxyproline level showed an inverse tendency during the increase of ALA concentration from 0.5 to 5.0 mmol/L.Conclusions Proper intensity of photodynamic reaction induced by ALA could enhance the secretion of MMP and inhibit the synthesis of collagen by skin fibroblasts,however,high concentration of ALA may exert an inverse effect.

Objective To investigate the association between genetic polymorphisms and protein levels of mannose-binding lectin (MBL) and the sensitivities of common infections in a pediatric Han population lived in Zhejiang Province.Methods MBL genetic polymorphisms of patients and controls were detected by PCR-based sequencing.MBL protein levels were measured using MBL ELISA Kit.Results No mutations at positions +223 and +239 of the exon 1 were detected in either patients or controls.No mutation at position +4 of the promoter was detected in controls.The frequencies of the three genotypes HH,HL,and LL at position-550 of the promoter were different between patients and controls(P＜0.05).The frequencies of genotypes YA and XB relevant to MBL protein levels were also different between patients and controls(P＜0.05).Comparing the frequencies of genotypes YA and XB in separate infectious disease with controls,significant differences were found in the group of RRI and CMV infection.The distributions of serum MBL level frequencies in patients and controls were both characterized by skewed distributions.MBL levels of patients with CMV infection were lower than those of controls(P＜0.05).Inversely,MBL levels of patients with acute respiratory infection and localized abscess were higher than those of controls (P＜O.05).Conclusion Genetic polymorphism of MBL gene is seemed to be relative to the sensitivity of common infections in children.

Objective To investigate the effect of intralesional lauromacrogol therapy on infantile hemangioma.Methods 30 cases all received intralesional injection of lauromacrogol.The changes of tumor size,texture and color were monitored and recorded.The adverse effects after medication were observed and managed accordingly.The adverse effects after medication were observed.The times of treatment were determined according to degree of reduction of tumor volume.To assess the efficacy,a 4 scales system was adopted.Results All patients had completed the treatment and followup.The overall response was scale Ⅰ in 2 cases (6.7 ％),scale Ⅱ efficacy in 4 patients (13.3 ％),scale Ⅲ efficacy in 5 cases (16.7 ％) and scale Ⅳ in 19 cases (63.3 ％).The smaller the tumor,the fewer the times of treatment,and the better the results.The effects of treatment were better if patient could be treated by intralesional lauromacrogol therapy in early stage.No severe adverse effects were observed during 6 months of follow-up.Conclusions Intralesional injection of lauromacrogol sclerotherapy is a safe and effective way to treat infantile hemangioma.

Objective To evaluate the clinical effectiveness of 5-aminolaevulinic acid (ALA)-based photodynamic therapy (PDT) for the treatment of moderate to severe acne vulgaris.Methods From January to June 2013,a total of 43 patients with moderate to severe acne vulgaris were treated by 5 aminolaevulinic acid based PDT with red light.All patients were received three times of treatment at 2-week intervals.Clinical assessment was conducted before and at 2,4,6 and 8 weeks after treatment.Results The lesion counts of both inflammatory and non-inflammatory lesions were reduced significantly after treatment.The mean percentage reduction in inflammatory lesions was 88.9 ％ at 8 weeks after the treatment,meanwhile that in non-inflammatory lesions was 73.4％.After 8 weeks of treatment,55.8％ patients (24/43)showed clinical cure,and 41.9％ patients (18/43)showed excellent response and 2.3％ patient (1/43) showed good response.Only 4.7％ patients (2/43)showed signs of recurrence after 4 months after ALA-PDT.After one course of ALA-PDT,the symptoms in this recurrent case were significant improvement.The common adverse effects included pain,edema and transient hyperpigmentation.They could gradually disappear without a need of special intervention.Conclusions ALA-PDT is a safe and effective therapeutic option for the treatment of moderate to severe acne vulgaris.

Objective:To investigate the effects of the Paecilomyces Lilacinuson extracellular polysaccharides on the phenotypic and maturation of murine dendritic cells. Methods: Imature DCs were induced in vitro from the murine bone marrow cells in the presence of rmGM-CSF and rmIL-4, and then they were cultured with different dosage of the extracellular polysaccharides. The morphological characterization was analyzed under microscopy. The expressions of the DCs surface costimulating factors and phagocytic function to FITC-dextran were detected by flow cytometry. The level of IL-12 secreted by DCs was observed by ELISA. At the same time the influence of DCs on the proliferation of T cells was determined by MTT. Results:Treating with the polysaccharides for 48 h could up-regulate the expression of DCs surface molecules,such as CD11c,MHCⅡ,CD80 and CD86,and the 400 μg/ml was the optimal dose,comparing with the blank control group, the difference was significant (P<0. 01), contrast to LPS control group that was not different ( P<0. 05 ) . The uptaking FITC-dextran ability of the DCs treated with 300 μg/ml and 400 μg/ml polysaccharides was significant lower than the unstimulated DCs(P<0. 05). At the same time the extract at different concentration could distinctly enhance the proliferation of T cells by DCs too. Conclusion:The extracellular polysaccharides could stimulate the maturation of dendritic cells and induce the production of mature dendritic cells.

Objective To evaluate the effect of sucralfate and acid-suppressive drugs on preventing ventilator-associated pneumonia (VAP) in mechanically ventilated patients.Methods All randomized controlled trials (RCTs),which studied the effect of sucralfate and acid-suppressive drugs on the incidence of VAP in mechanically ventilated patients,were searched from PubMed,Embase and the Cochrane Library during January 1966 to March 2013 via manual and computer retrieval.All related data were extracted.Meta analysis was conducted using the statistical software RevMan 5.2 and the quality of the RCTs was strictly evaluated with the methods recommended by the Cochrane Collaboration.Results A total of 15 RCTs involving 1315 patients in the sucralfate group and 1568 patients in the acid-suppressive drug group were included in this study.The incidence of VAP was significantly reduced in the sucralfate group (RR =0.81,95％ CI 0.7-0.95,P =0.008),while no difference was found between the two groups in the incidence of stress-related gastrointestinal bleeding (RR =0.96,95％ CI 0.59-1.58,P =0.88).No statistical difference was found in the days on ventilator,duration of ICU stay and ICU mortality in the two groups (all P values ＞ 0.05).Conclusion In patients with mechanical ventilation,sucralfate could decrease the incidence of VAP,while has no such effect on the stress-related gastrointestinal bleeding,the days on ventilator,duration of ICU stay and ICU mortality.

Objective To investigate the anti-oxidant effect of pirfenidone (PD) at different dosage on acute lung injury (ALI) induced by paraquat (PQ) poisoning in mice. Methods 144 ICR mice were randomly divided into four groups: control group (n = 24), PQ poisoned group (n = 24), high and low doses PD treatment groups (n = 48). ALI induced by PQ poisoning model was reproduced by intraperitoneal injection of 25 mg/kg 20% PQ solution in mice, and the mice in control group was given equal volume of normal saline. Intragastric administration with 30 mg/kg and 70 mg/kg PD suspension [PD was dissolved in 0.4% sodium carboxymethyl cellulose sodium (CMC) solution] after PQ poisoning immediately for 3 days in high and low doses PD treatment groups respectively, while the same volume of 0.4% CMC solution was administrated in control group and PQ poisoned group. Then mice in each group were respectively sacrificed at 2, 6, 12, 24, 48 and 72 hours after PD exposure to harvest the lung tissue, nuclear factor-κB (NF-κB) was determined by enzyme linked immunosorbent assay (ELISA), superoxide dismutase (SOD) and malonaldehyde (MDA) were determined by colorimetry, and pulmonary pathological changes were observed with microscope after hematoxylin-ensin (HE) staining. Results Compared with the control group, NF-κB from 2 hours in PQ poisoned group was significantly increased (pg/mg: 106.65±5.96 vs. 79.04±2.40, P < 0.05), and lasted to 72 hours (pg/mg: 110.47±5.91 vs. 82.70±2.79, P < 0.05); the activity of SOD was significantly enhanced in early stage (2-6 hours; U/mg: 39.34±1.17 vs. 34.72±1.54 at 2 hours, 37.37±0.90 vs. 33.75±0.93 at 6 hours, both P < 0.05) followed by a gradual decrease; the content of MDA within 24 hours was significantly increased (nmol/mg: 1.67±0.22 vs. 1.03±0.09 at 2 hours, 1.56±0.17 vs. 1.14±0.16 at 24 hours, both P < 0.05) followed by a gradual decrease. Compared with the PQ poisoned group, both high and low dose PD treatment could significantly inhibit NF-κB from 24 hours to 72 hours, and significantly inhibit MDA within 24 hours; high dose PD treatment could increase SOD activity at 6 hours, which showed a tendency of decreasing followed by increasing in low dose PD treatment group. Compared with high dose PD treatment group, the inhibition of MDA in low dose PD treatment group was more significant (nmol/mg: 0.90±0.08 vs. 1.29±0.18 at 2 hours, 1.03±0.32 vs. 1.84±0.43 at 6 hours, 1.08±0.09 vs. 1.33±0.16 at 24 hours, all P < 0.05), and SOD activity was significantly decreased at 6 hours (U/mg: 35.24±2.08 vs. 38.46±0.87, P < 0.05), and it was increased at 72 hours (U/mg: 39.81±1.30 vs. 34.58±3.15, but P > 0.05), but no significant difference in NF-κB activity at all time points was found. Under light microscope, a wide range of red blood cells and serous effusion, alveolar septum fracture and pulmonary interstitial inflammatory cell infiltration were shown by pathologic examination in PQ poisoned group. The pathologic changes in high and low doses PD treatment groups were obviously less than those of PQ poisoned group, and no significant difference was found between the two doses groups. Conclusions The early therapeutic effect of PD may relate to the inhibition of NF-κB and reactive oxygen species, then reduce the inflammation of PQ poisoning. The treatment effectiveness of low dose PD seems better than high dose PD.

BACKGROUND: Metoprolol is a selective β1-Blocker commonly used in essential hypertension. It is metabolized by CYP2D6. CYP2D6*10, which was identified to decrease activity of CYP2D6, is the main variance in Chinese population. β1-adrenergic receptor, with Ser49Gly and Gly389Arg polymorphisms, is the target of metoprolol. It was still unknown that whether the CYP2D6 and β1-adrenergic receptor had a synergic effect on metoprolol antihypertension therapy. AIM: To clarify the genetic polymorphism associated with metoprolol pharmacokinetics and pharmacodynamics in antihypertension therapy. METHODS: 125 mild-to-med essential hypertension patients were enrolled in this study. Patients were mono-therapied with metoprolol for 12 weeks. Blood pressure was monitored every 4 weeks. PCR-RFLP method was use to identify CYP2D6*10 and β1-adrenergic receptor Ser49Gly and Gly389Arg polymorphisms. Plasma metoprolol concentration was measured by HPLC- fluorescence detection. RESULTS: Trough blood level (C0) of metoprolol was associated with CYP2D6*10 variance in a gene-dose-effect manner, whereas the extent of blood pressure decrease was not significant different in CYP2D6*1*1, *1*10 and CYP2D6*10*10 patients. After 12 weeks metoprolol therapy, Gly49 carriers had stronger decrease in systolic and diastolic blood pressure than that of Ser49 homozygotes. Similarly, subjects homozygous for Arg389 had stronger decrease in blood pressure than that of Gly389 carriers. CONCLUSION: CYP2D6*10 variance significantly change the pharmacokinetics of metoprolol, and the genetic polymorphisms of β1-adrenergic receptor were associated with the pharmacodynamics of metopolol in antihypertension therapy.

Objective To investigate the effects of Paecilomyces Lilacinuson extracellular polysaccharides on the phenotypic and function maturity of mouse dendritic ceils.Methods Mononuclear cells were isolated from the mouse bone marrow cavity and added with cytokines for obtaining the recombinant mouse granulocyte-macrophagocyte colony stimulating factor(rmGM-CSF),recombinant mouse interleukin 4(rmIL-4) was induced to differentiated to immature DCs.Then different concentrations of extracellular polysaccharides were used to conduct the intervention.The mature DCs surface marker CD11c,major histocompatibility complex Ⅱ (MHC Ⅱ),CD80,CD86 molecular expression and phagocytosing FITC-dextran ability was detected by the flow cytometry.The effect of the polysaccharides on DCs Toll-like receptor(TLR)2 mRNA and TLR4 mRNA expression was detected by RT-PCR.Results After 400 μg/mL polysaccharides action for 48 h,the expression of DCs surface molecules such as CD11c,MHC Ⅱ,CD80 and CD86 was significantly up-regulated compared with the blank control group (P＜0.05);after the polysaccharides action,the ability of DCs phagocytosing FITC-dextran was decreased,especially the effects of 300,400 μg/mL of polysaccharides were more significant compared with the control group (P＜0.05).In addition,the polysaccharides could down-regulate the expression of TLR2 mRNA and TLR4 mRNA in DCs,the DCs down-regulation effect after 100-400 μg/mL polysaccharides treatment,the difference compared with the blank control group was statistically significant(P＜0.05).Conclusion The extracellular polysaccharides can up-regulate the expression of DCs surface CD11c,MHC Ⅱ,CD80 and CD 86 molecules,decreases the phagocytosis ability and down-regulates the expression of TLR2 mRNA and TLR4 mRNA,which preliminarily indicates that the polysaccharides could stimulate the differentiation and maturation of murine DCs.

BACKGROUND: There is a growing recognition that the adipose tissue is an endocrine organ that secretes signaling molecules such as adiponectin and resistin. The peroxisome proliferator activated receptor γ (PPARγ) is expressed in high levels in the adipose tissue. Thiazolidinediones are selective PPARγ agonists with insulin-sensitizing properties. It has been postulated that thiazolidinediones such as rosiglitazone exert their pharmacodynamic effects in part through modulation of resistin (implicated in insulin resistance) and adiponectin (an insulin-sensitizing molecule) expression subsequent to activation of PPARγ. There are conflicting data, however, on the biological direction in which resistin expression is modulated by PPARγ agonists and whether an increase in adiponectin expression can occur in the face of an upregulation of resistin. METHODS: Using the murine 3T3-L1 adipocytes as a model, we evaluated the changes in resistin and adiponectin gene expression after vehicle, rosiglitazone (10 μmol/L, a PPARγ agonist), GW9662 (5 μmol/L, a selective PPARγ antagonist) or GW662 and rosiglitazone co-treatment.RESULTS: In comparison to vehicle treatment, rosiglitazone increased the average adiponectin and resistin mRNA expression by 1.66- and 1.55-fold, respectively (P＜0.05). Importantly, GW9662 also upregulated adiponectin expression (by 1.57-fold, P＜0.05) but did not influence resistin expression (P＞0.05). Co-treatment with rosiglitazone and GW9662 maintained the adiponectin upregulation (1.87-fold increase from vehicle, P＜0.05) while attenuating resistin upregulation (1.31-fold increase from vehicle, P＜0.05) induced by rosiglitazone alone (1.55-fold increase from vehicle, P＜0.05). CONCLUSION: This study presents new evidence that adiponectin transcript is upregulated with both a PPARγ agonist (rosiglitazone) and antagonist (GW9662), while GW9662 co-treatment does not block rosiglitazone-induced adiponectin upregulation. These data collectively suggest that biological mechanisms independent from PPARγ may underlie thiazolidinedione pharmacodynamics on adiponectin expression. Moreover, increased adiponectin expression by GW9662, in the absence of an upregulation of resistin expression, lends further support on the emerging clinical potential of PPARγ antagonists in treatment of insulin resistance. Decreased resistin expression may not be crucial for the insulin-sensitizing effect of rosiglitazone. These findings may serve as a foundation for future dose-ranging and time-course studies of thiazolidinedione pharmacodynamics on adipocytokine expression in human adipocytes.