Objective To establish an animal model for immunological contact urticaria in mice.Methods A total of 60 BALB/c mice were randomly and equally divided into 5 groups:anti-dinitrophenol IgE monoclonal antibody (anti-DNP IgE) + 2,4-dinitrofluorobenzene (DNFB) group and anti-DNP IgE + trimellitic anhydride (TMA) group both injected with anti-DNP IgE via tail veins firstly,followed by topical treatment with DNFB and TMA respectively on the ears at 24 hours after the injection,DNFB group,TMA group and normal saline (NS) group all injected with NS via the tail vein firstly,followed by topical treatment with DNFB,TMA and NS on the ears 24 hours after the injection.In the following 14 days,mice were observed daily for the appearance of wheals and for scratching behavior.All the mice were sacrificed at the end of the study followed by determination of the percentage of degranulated mast cells and spleen index as well as observation of pathological changes.Results Wheals were observed in all the mice (12/12) in the anti-DNP IgE + DNFB group,some mice (8/12) in the anti-DNP IgE + TMA group,but not observed in any mice in the other 3 groups.Compared with the NS group,both the anti-DNP IgE + DNFB group and anti-DNP IgE + TMA group showed a significant increase in the percentage of degranulated mast cells (70.21％ ± 26.01％ and 54.25％ ± 39.57％ vs.14.45％ ±6.79％,F=14.41,P=0.000),spleen index (7.54 ± 1.56 and 7.87 ± 1.18 vs.5.37 ± 1.16,F=4.29,P=0.004) and scratching frequency ((31.58 ± 3.58)/h and (22.17 ± 3.81)/h vs.(2.00 ± 0.85)/h at 30 minutes,F =437.86,P ＜ 0.01).Conclusion A stable mouse model for immunological contact urticaria can be established quickly by sensitization with anti-DNP IgE and challenge with DNFB.

Objective To determine the coagulation status as well as circulating levels of complement and inflammation markers in patients with chronic urticaria (CU) during acute attack and in remission,and to estimate the relationship of coagulant and anticoagulant factors as well as fibrinolytic markers with the development of chronic urticaira.Methods This study included 40 patients with CU (22 during acute attack and 18 in remission) and 40 healthy blood donors from the Guangzhou Blood Center.Venous blood samples were obtained from these subjects,and enzyme-linked immunosorbent assay (ELISA) was performed to measure the plasma levels of prothrombin fragrnent 1 +2 (F1 +2),tissue factor (TF),thrombomodulin (TM),high molecular weight kininogen (HMWK),tissue-type plasminogen activator (t-PA),C5a and serum levels of C3,C4,antistreptolysin O antibodies (ASO),rheumatoid factor (RF) and C-reactive protein (CRP).Erythrocyte sedimentation rate (ESR) was also determined in these patients.Comparisons of these parameters were carried out by using t test,and the correlation of these factors with CU was evaluated by using Spearman correlation coefficient.Results Compared with the healthy controls,the patients with CU showed significantly higher plasma levels of F1+2 and HMWK (both P ＜ 0.01),but lower levels of TF,TM and t-PA (all P ＜ 0.01).The plasma levels of F1 +2,HMWK,t-PA were significantly correlated with the symptom scores in patients with CU (r =0.81,P ＜ 0.01; r =-0.39,P ＜ 0.05; r =0.35,P ＜ 0.05).A significant increase was observed in the plasma concentration of F1 +2 in patients during acute attack compared with those in remission (P ＜ 0.01),whereas no significant differences were noted in the plasma levels of TF,TM,HMWK,t-PA,C5a,serum levels of C3,C4,ASO,RF and CRP or ESR between the two groups of patients (all P ＞ 0.05).Conclusions It seems that coagulation,anti-coagulation and fibrinolysis are all involved in the development of urticaria.There is an obvious difference in the plasma level of prothrombin F1 +2 between patients with CU during acute attack and in remission,suggesting that coagulation factors play a certain role in the initiation and progression of CU.

Objective To investigate the role of regulatory T (Treg) / T helper type 17 (Th17) cells in the pathogenesis of chronic spontaneous urticaria (CSU).Methods Eighty-nine patients with CSU were enrolled in this study,including 48 in active stage and 41 in remission stage.Forty-eight health check-up examinees,who were collected from the community hospitals in Guangzhou city,served as the healthy controls.Fluorescence-based realtime quantitative PCR was performed to determine the expression of transcription factors FOXP3 and RORγt in PBMCs from these subjects.Results Compared with the patients with CSU in remission stage and healthy controls,the patients in active stage showed a significantly higher level of FOXP3 mRNA (0.57 ± 0.19 vs.0.11 ± 0.21 and 0.13 ± 0.23,both P ＜ 0.05),but a significantly lower level of RORγt mRNA (0.43 ± 0.39 vs.0.89 ± 0.40 and 0.87 ± 0.43,both P ＜ 0.05).Conclusions The expression of Treg cell regulator FOXP3 increases,while the expression of Th17 cell regulator RORγt decreases in patients with CSU,suggesting that the imbalance between Treg and Th17 cells induced by the interaction between FOXP3 and RORγt may be involved in the pathogenesis of CSU.

Objective To construct human canstatin gene eukaryotic expression vector and investigate the therapeutic effect of intramuscular canstatin gene delivered by electroporation on tumor growth.Methods Canstatin cDNA was amplified from total RNA extracted from fresh fetal liver by reversing transcription polymerase chain reaction (RT-PCR).The canstatin cDNA fragment was in serted into pEGFP-N1 eukaryotic expression vector.The recombination plasmid was delivered to the quadriceps of the mice with Lewis lung carcinomas by electroporation intramuscular.Fluorescence intension measured by fluorescence microscope,reverse-PCR assay,and immunohistochemistry assay were performed to detect the expression of canstatin gene in the muscle and in circulation.The tumor weight and volume were used to detect the biological effects of canstatin gene delivery.Results Recombinant eukaryotic expression vector of recombinant human canstatin was successfully constructed.The canstatin mRNA was significantly increased in the skeletal muscle and intramuscular delivery of canatatin gene by electroporation acquired the expression of enhanced green fluorescent protein (EGFP)/canstatin protein in the circulation and significantly inhibited tumor growth.The percent of the inhibition of tumor weight was 57.7 ％.Conclusions Electroporation mediated gene transfer efficiency in skeletal muscle was compared to simple plasmid injection and lasted for a long time.It was an efficient and safe,convenient and economic,gene transfer methods and might have certain clinical application value.Electroporation mediated canstatin gene transfer in skeletal muscle had obvious inhibitory effect on Lewis lung cancer in mice subcutaneous xenograft tumor growth.

Objective To study the influence of sunscreens with different efficacy on delayed type hypersensitivity (DTH) and their immunoprotective effect in mice.Methods A cohort of mice were randomly divided into 5 groups with 10 mice in each group:group 1 as the positive control without irradiation,group 2 receiving solar-simulated radiation (SSR) only,group 3 receiving SSR and protected by sunscreen l with sun protection factor 15(SPF15)and persistent pigment darkening(PPD)12,group 4 receiving SSR and protected by sunscreen 2 with SPF 50 and PPD 28,and group 5 as the negative contml receiving SSR only.SSR was carried out on the back of mice with the UVA dose being 1.4 J/cm2 and UVB dose being 100 mJ/cm2 for 10 days.After a 5-day irradiation,the groups 1 to 4 were immunized by intraperitoneal injection with 100 μl(107 cells/ml) of Candida albicans suspension.On the 10th day both sides of the posterior foot pad were measured;then the foot pads were injected with additional 50 μl of the Candida albicans suspension.Twenty-four hours after the injection,the thickness of each foot pad was measured,and immunosuppression rate was calculated.Finally,the mice were sacrificed and skin samples were obtained from the back of these mice followed by the examination of CDla, CD80 and CD86 expression by Western blot.Resets The thickness of edema in foot pads was 0.41±0.38 mm,0.21±0.23 mm and 0.30 ± 0.25 mm in group 1,3 and 4,respectively,significantly higher than in group 5 and 2(0.04±0.03 mm,0.14±0.12 mm,respectively,all P<0.05),while no significant difference was observed between the group 3 and 4(P>0.05).Significant differences were observed in the immunosuppression rate between group 2,3 and 4(73.0%±11.3%,54.1%±6.4%,29.7%±7.5%,respectively,all P<0.01).Western blot revealed a significant increment in the expression of CDla protein in group 1 compared with group 2 as well as in the expression of CD86 protein in group 1 and group 3 compamd with group 2 and group 5(all P<0.05),but no statistical difference was observed between the other groups in the expression level of CDla,CD80 or CD86(P>0.05).Conclusions The exposure to sub-erythema dose of UV can induce DTH,and sunscreens have an immunoprotective effect in this process.Epidermal Langerhans cells are not essential for UV-induced immunosuppression.

Objective To evaluate the relationship of clinical symptom to plasmic levels of D-dimer, activated factorⅦ (FⅦa) and tissue factor pathway inhibitor (TFPI)/X a in patients with urticaria. Methods A total of 27 patients with chronic urticaria (CU), 27 patients with acute urticaria (AU) and 26 normal human controls were included in this study. Symptom score was determined and disease course was surveyed in these patients. ELISA was used to detect the plasma levels of D-dimer, FⅦa and (TFPI)/Xa in patients and controls. The relation of clinical symptom and disease course to plasma levels of these parameters was assessed. Results In patients with AU and normal controls, the plasma level of D-dimer was 450.57± 242.13 ng/mL and 266.81±40.68 ng/mL, respectively, the level of FⅦa, 2.23± 0.74 ng/mL and 5.23±1.35 ng/mL, respectively, and the level of TFPI/Xa 0.87±0.13 nmol/L and 0.88 ~ 0.12 nmol/L, respectively. There was a significant difference in the level of both D-dimer and FⅦa (both P < 0.01 ), whereas no differ-ence was observed in that of TFPI/X a (P > 0.05) between patients with AU and normal controls. In addi-tion, increased level of D-dimer and decreased level of FⅦa were noticed in patients with CU compared with those in normal controls (593.80±294.04 ng/mL vs 266.81±40.68 ng/mL, 3.98±0.35 ng/mL vs 5.23± 1.35 ng/mL, both P < 0.01 ), but there was no significant difference in the plasma level of TFPI/Xa (0.87± 0.16 nmol/L vs 0.88±0.12 nmol/L, P > 0.05). Significant difference was observed in the plasma level of D-dimer and FⅦa between patients with AU and CU (450.57±242.13 ng/mL vs 593.80 ±294.04 ng/mL, P < 0.05; 2.23± 0.74 ng/mL vs 3.98± 0.35 ng/mL, P<0.01 ). The plasma level of D-dimer positively corre-lated to the symptom score of patients with CU and those with AU (r= 0.68, P< 0.01; r= 0.82, P< 0.01),but was independent of discase course (P> 0.05). Neither the level of FⅦa nor that of TFPI/Xa correlated to symptom score or disease course of patients (all P > 0.05). Conclusions There is an overactivation of coagulation cascade, consumption of blood coagulation factors and secondary fibrinolysis in patients with urticaria, suggesting that plasma D-dimer and FⅦa may be associated with the clinical symptoms of urticaria.

Objective To evaluate effects of tea polyphenols on the mRNA and nucleoprotein expression of Nrf2/Bach1 in human skin fibroblasts (HSFs).Methods Some HSFs were incubated with tea polyphenols at different concentrations of 0,2.5,5,10,20 and 40 mg/L for 24 hours.Methyl thiazolyl tetrazolium (MTT) assay was conducted to evaluate the proliferative activity of HSFs to screen the optimal concentration of tea polyphenols.Then,some other HSFs were treated with tea polyphenols at this optimal concentration for 24 hours.Real-time quantitative PCR (RT-qPCR) was performed to determine mRNA expression of Nrf2 and Bach1,Western blot analysis to measure nuclear expression of Nrf2 and Bach1 proteins,and immunofluorescence assay to determine the distribution of Nrf2 and Bach1 protein in the cell nucleus.Results MTT assay showed that 5 mg/L tea polyphenols had no obvious effects on the proliferation of HSFs,so 5 mg/L was chosen as the optimal concentration of tea polyphenols for subsequent experiments.HSFs cultured without tea polyphenols served as control group.After the treatment,the 5-mg/L tea polyphenol group showed significantly decreased mRNA and nuclear protein expression of Bach 1 (mRNA:0.629 ± 0.077 vs.0.940 ± 0.033,t =6.397,P ＜ 0.05;protein:1.424 ± 0.171 vs.16.966 ± 1.702,t =15.730,P ＜ 0.05),but significantly increased mRNA and nuclear protein expression of Nrf2 (mRNA:1.467 ± 0.076 vs.0.977 ± 0.091,t =7.133,P ＜ 0.05;protein:6.929 ± 0.121 vs.3.537 ± 0.126,t =33.636,P ＜ 0.05) compared with the control group.Immunofluorescence assay showed increased accumulation of Nrf2 protein,but decreased accumulation of Bach1 protein in the nucleus.Conclusion Tea polyphenols can promote the mRNA and nuclear protein expression as well as nuclear distribution of Nrf2,but suppress the mRNA and nuclear protein expression as well as nuclear distribution of Bach 1,finally exerting antioxidative effects.

Objective To evaluate the scavenging effect of crude polysaccharides extracted from Lycium barbarum (LBP) on reactive oxygen species in ultraviolet radiation-induced HaCaT cells,and to explore its possible mechanism.Methods Cultured immortalized human keratiuocyte HaCaT cells were divided into 6 groups:blank control group receiving no treatment,LBP group treated with crude LBP alone,ultraviolet A (UVA) group treated with UVA radiation alone,ultraviolet B (UVB) group treated with UVB radiation alone,UVA + LBP group treated with crude LBP for 24 hours followed by UVA radiation,and UVB + LBP group treated with crude LBP for 24 hours followed by UVB radiation.MTT colorimetry was performed to evaluate the cellular proliferative activity,UV spectrophotometric method to measure the UVA and UVB absorption of crude LBP,dichlorodihydrofluorescein diacetate (DCFH-DA) fluorescent probe assay to detect the level of ROS,enzymatic-biochemical method to estimate the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px),as well as to detect the leakage of lactate dehydrogenase (LDH).Results Crude LBP at different concentrations of 0,100,200,300,400,500,600,1 500,2 000 mg/L had no obvious effects on the proliferative activity of HaCaT cells.Crude LBP had a high transmittance of ultraviolet rays at 280-400 nm.Compared with the blank control group,the UVA group and UVB group both showed significantly higher LDH leakage and ROS level,lower activities of SOD and GSH-Px (P ＜ 0.001 or 0.05).Pretreatment with crude LBP before the ultraviolet radiation could significantly increase the activities of SOD and GSH-Px,decrease the LDH leakage and ROS level in the UVA + LBP group and UVB + LBP group compared with the UVA group or UVB group (P ＜ 0.05).Conclusion Crude LBP have no effect of sunscreening agents,but can effectively scavenge ROS,decrease LDH leakage,inhibit ultraviolet radiation-induced photodamage in HaCaT cells,which may be associated with the enhancement of antioxidant enzyme activity.

Objective:To observe the effect of observing good swallowing on the swallowing action of stroke survivors with dysphagia.Methods:Eighteen stroke survivors with dysphagia were randomly divided into a treatment group ( n=9) and a control group ( n=9). In addition to routine swallowing rehabilitation therapy, the treatment group was asked to simulate swallowing after watching a video of normal people′s swallowing action. They did so 5 times a week for 10 minutes, while the control group just watched landscape videos at the same time. The treatment lasted 8 weeks. Before and after the treatment, both groups were assessed using the eating assessment tool (EAT-10), the functional oral intake scale (FOIS) and the penetration and aspiration scale (PAS). Functional magnetic resonance imaging (fMRI) was also used to observe their swallowing action. Results:There was no significant difference between the two groups in any of the measurements before the treatment. After the 8 weeks of treatment the average EAT-10, FOIS and PAS scores of the treatment group were all significantly better than before the treatment and better than the control group′s averages at the time. fMRI showed significantly more areas activated in the precuneus, parietal lobe, posterior central gyrus, BA7, BA5, frontal lobe and paracentral lobule in the treatment group compared with before the intervention and also more than in the control group.Conclusions:Observing proper swallowing action can improve dysphagia and activation of the swallowing-related brain areas of stroke survivors.

Objective To systematically evaluate the effects of mechanical chest compression (CC) combined with manual CC and single-manual CC on the outcome indexes of cardiopulmonary resuscitation (CPR) for patients with in-hospital cardiac arrest (IHCA). Methods The relevant publicly published literatures about the effects of mechanical CC combined with manual CC and single-manual CC on the outcome of CPR were searched by using the following Chinese keywords for retrieval: "cardiac arrest, asystole, sudden death, artificial recovery, artificial press, artificial CC, unarmed CPR, unarmed resuscitation, unarmed compressions, unarmed chest compressions, unarmed, artificial, resuscitation instrument, resuscitation machine, resuscitator, CPR, LUCAS, Autopulse, Thumper, MSCPR-1A"in databases such as China Biomedical Literature (CBM), VIP, Wanfang, and China National Knowledge Internet (CNKI) from their dates of foundation to March 11, 2019, and using the following key words in English "heart arrest, cardiac arrest, cardiopulmonary arrest, Cardiopulmonary Resuscitation, Resuscitation, Cardio-Pulmonary Resuscitation, CPR, compression, mechanical, automatic, automated, load distributing band, LBD, Autopulse, LUCAS" to retrieve all the published articles especially concerning the topics on the application effects of mechanical combined with manual CC for IHCA patients' CPR in the America National Library database (PubMed), Excerpta Medica (EMbase), Web of Science, and Cochrane Library from the establishment of the databases to March 11, 2019. The indexes of outcomes included return of spontaneous circulation (ROSC) rate, survival rate after hospital discharge and incidence of complications. The literatures were extracted independently by two reviewers, the qualities of the included randomized controlled trials (RCTs) were evaluated according to the Cochrane bias risk assessment tool, and the qualities of the included observational studies were evaluated according to the literature quality assessment form (NOS). Meta analysis was performed by using RevMan 5.3 software, and publication bias was assessed by using funnel plot. Results Twenty-one studies were enrolled, including 11 RCT articles and 10 observational studies; there were 2 005 participants. The results of this Meta-analysis showed that compared with manual CC, the ROSC rate and after discharge survival rate of IHCA patients were obviously higher in combined CC group [ROSC: odds ratio (OR) = 2.50, 95% confidence interval (95%CI) = 2.03-3.09, P < 0.000 01; discharge survival rate: OR = 2.71, 95%CI = 1.91-3.85, P < 0.000 01]; the incidence of complications of combined CC was lower than that in single manual CC (OR = 0.30, 95%CI = 0.13-0.68, P = 0.004). The funnel plots indicated that there was no apparent bias in the ROSC; because the enrolled studies were relatively few, it was difficult to evaluate the symmetrical characteristics of the funnel plots for discharge survival rate and the complication rate. Conclusions For IHCA patients, combined CC can improve ROSC, discharge survival rate, and reduce the occurrence of complications. It is suggested that during the actual rescue of IHCA patients, it is better to use combined CC, that is to say, manual CC should be adopted immediately in the early stage and then replace the mechanical CC device as soon as possible.