Aim DNA vaccinating BALB/c mice with the constructed recombinant plasmid, pcDNA3, containing ROP1 gene from Toxoplasma gondii to observe the effect on the production of the cytokines, IFN- γ、 IL - 2 , and NO in the immunized mice. Methods Large-scale preparation of plasmid DNA by alkaline lysis,the DNA were injected through muscles of left leg in each mouse at the dosage of 100μg. A booster vaccine was given at the same dosage after two weeks. Control groups were injected with pcDNA3 blank plasmid and normal saline respectively. After 30,50 and 70 days of the booster injection, following tests were carried out 3 times separately :the serum IFN-γand IL-2 were detected by sandwich ELISA;the NO was detected by enzyme assay. Results The 3 times detected results of IFN-γγ、IL-2 and NO were significant higher in the vaccinated group than that of in control groups and the contents were increased with the vaccinated time prolonged. Conclusion The IFN - γγ、 IL - 2 and NO were produced by vaccinating BALB/c mice with the recombinant plasmid, pcROP1.

Objective To evaluate the effect of calcitonin gene-related peptide (CGRP) on anoxiareoxygenation (A/R)-induced injury to neonatal rat cardiomyocytes incubated in high glucose medium.Methods Cardiomyocytes were obtained from 1-3-day old Sprague-Dawley rats and cultured in the culture medium containing 15％ bovine calf serum and then seeded onto 6-well plates at a density of 10 × 105/ml (3 ml/well).The cells were randomly divided into 5 groups (n =9 each):normal glucose medium control group (NG group),high glucose medium group (HG group),high glucose medium + A/R group (HG+ A/R group),high glucose medium + A/R + CGRP group (HG + A/R + CGRP group),and high glucose medium + A/R + CGRP+ CGRP8-37 group (HG + A/R + CGRP + CGRP8-37 group).The cells were incubated in normal glucose (5.5 mmol/L) medium for 72 h in NG group.In HG group,the cells were incubated in high glucose (25.0 mmol/L) medium for 72 h.In HG + A/ R group,the cells were incubated in high glucose medium for 72 h and then exposed to 3 h of anoxia followed by 2 h of reoxygenation.In group HG + A/R + CGRP,the cells were incubated in high glucose medium for 72 h,CGRP (final concentration 10-8 mol/L) was then added to the culture media and the cells were incubated for 1 h and then underwent A/R.In HG + A/R + CGRP + CGRP8-37 group,the cells were incubated in high glucose medium for 72 h,CGRP (final concentration 10 8 mol/L) was then added to the culture media and the cells were incubated for 1 h and then underwent A/R.In HG + A/R + CGRP + CGRP8-37 group,the cells were incubated in high glucose medium for 72 h,CGRP8-37 (final concentration 10-8 mol/L) and CGRP8-37 (CGRP receptor antagonist,final concentration 10-7 mol/L) was then added to the culture media and the cells were incubated for 1 h and then underwent A/R.Apoptosis in cardiomyocytes was detected using TUNEL and apoptosis index (AI) was calculated.The lactate dehydrogenase (LDH) activity in the culture medium was analyzed.Results AI and LDH activity were significantly higher in HG group than in NG group,and in HG + A/R group than in HG group.Compared with HG + A/R group,AI and LDH activity were significantly decreased in HG + A/R + CGRP group,while no significant changes were found in HG + A/R + CGRP + CGRP8-37 group.Compared with HG + A/R + CGRP group,AI and LDH activity were significantly increased in HG + A/R + CGRP + CGRP8-37 group.Conclusion CGRP attenuates A/R-induced injury to neonatal rat cardiomyocytes incubated in high glucose medium via combing with CGRP receptor.

Objective:To screen for peptides that specifically bind to PreS1 antigen from a phage-displayed peptide library.Methods: The PreS1 antigen was used as the target molecule to screen the binding peptide from the Ph.D.-7 peptide library with phage display technique,and the positive clones were identified by ELISA.Results: After three rounds of biopanning,the binding peptides were screened from the peptide library by ELISA and competitive inhibiting ELISA.Sequencing result showed that the binding peptides had high affinity and specificity.Conclusion: A peptide binding PreS1 antigen has been successfully obtained by screening the phage display library,which paves a way for the diagnosis and treatment of hepatitis B infection.

Objective To assess the protective effect of mastoparan-1(MP-1) on acute lung injury in mouse model with endotoxemia(ETM) induced by lipopolysaccharide(LPS),and to investigate the possible mechanism of protcetive effect.Methods The endotoxemia murine model was reproduced by tail vein injection of LPS(5mg/kg) in mice.Animals were randomly divided into normal control group(n=8),endotoxemia group(n=48) and MP-1-treatment group(MP-1 was injected in 3mg/kg at the same time of LPS injection,n=48).Animals of the latter two groups were sacrificed at 2,6,12,24,48 and 72 hours after injection,and then the blood and lung tissue samples were collected.Plasma LPS was assayed using kinetic turbidimetric limulus test,TNF-? and IL-6 were measured by appropriate ELISA kits,TLR4,TNF-? and IL-6 mRNA expressions in lung tissues were analyzed by real-time RT-PCR,myeloperoxidase(MPO) activity of lung tissues was determined by spectrophotometric method,and the pathological changes in lung tissues were observed under microscope.Result The plasma levels of LPS,TNF-? and IL-6 in the mice of endotoxemia group were increased at 2-48 hours after LPS injection(P

Objective To study the influence of dihydromyricetin on the oxidation and non-enzymatic glycosylation in impaired glucose tolerance rats.Methods Animal model of impaired glucose tolerance rat was built by intragastrical(i.g.) injection of D-galactose generally.After 8 weeks intragastrical(i.g.) injection of dihydromyricetin(experimental group) and metformin (positive control),the levels of fasting blood glucose and tow-hour postprandial blood glucose,the insulin level and its resistance index,the levels of glycosylated hemoglobin (GHb) and advanced glycation end-products(AGEs) were detected,and the activities of superoxide dismutase(SOD) and glutathione peroxidase (GSH-Px),the content of malondiadehyde(MDA) were determined respectively.Results Compared with model group,dihydromyricetin decreased the level of two-hour postprandial blood glucose (P0.05).Conclusion Dihydromyricetin can significantly improve the state of impaired glucose tolerance rats; the mechanism may be related to its effect of improving the antioxidant activity of GSH-Px,inhibiting non-enzymatic glycation,lowering body oxidative stress and insulin resistance and promoting the use of blood glucose in peripheral organizations.

Objective To investigate the effects of substance P on anoxia/reoxygenation (A/R) injury to cardiomyocytes of neonatal rats.Methods Cardiomyocytes of neonatal rats were isolated from Sprague-Dawley rats,aged 1-3 days,and were cultured in 6-well plates for 72 h.The cardiomyocytes were then assigned into 4 groups (n =3 each) using a random number table:control group (group C),A/R group,substance P group (group SP) and substance P + D-SP group (group SP + D-SP).The cells were cultured routinely for 6 h in group C and the cells were exposed to 99.9 ％ N2 in an incubator at 37 ℃ for 3 h followed by reoxygenation for 2 h in the other groups.The cells were incubated with substance P with the final concentration of 10-7 mol/L for 1 h before anoxia in group SP.The cells were incubated for 1 h with substance P with the final concentration of 10-7 mol/L and D-SP (a specific antagonist of neurokinin-1 receptor) with the final concentration of 10-6 mol/L before anoxia in group SP + D-SP.The apoptosis rate and lactate dehydrogenase (LDH) activity were detected at the end of reoxygenation using TUNEL assay and LDH assay kit,respectively.Results Compared with C group,the apoptosis rate and LDH activity were significantly increased in A/R,SP and SP+ D-SP groups.Compared with A/R group,the apoptosis rate and LDH activity were significantly decreased in SP and SP + D-SP groups.Compared with SP group,the apoptosis rate and LDH activity were significantly increased in SP + D-SP group.Conclusion Substance P can attenuate A/R injury to cardiomyocytes of neonatal rats,and activation of neurokinin 1 receptors is involved in the mechanism.

The pathogenesis of icteric hepatitis is that pathogenic dampness blocks the middle energizer or blood stasis blocks the bile duct,so that the bile is not excreted normally and moves to the skin surface,which makes the skin yellow.Traditior al Chinese medicine (TCM) has unique advantages in the prevention and treatment of icteric hepatitis.This article systematically introduces the etiology and pathogenesis of icteric hepatitis and the TCM syndrome differentiation therapy for icteric hepatitis,in order to provide a reference for the clinical treatment of icteric hepatitis and the improvement in its prognosis.

Objectives To investigate the effect of tumor necrosis factor ? on skeletal muscle ischemia reperfusion injury (SMIR). Methods Twenty four healthy male Sprague Dawley rats were divided into three groups. Group A underwent anesthesia and external jugular vein cannulation only. Group B underwent 4 hours of left hind limb ischemia followed by 4 hour reperfusion. Group C ischemia and reperfusion were treated with anti TNF ? monoclonal antibody (2?mg/kg). ResultsSMIR significantly increased the transcription of TNF ? mRNA in monocyte. The increased TNF ? raised significantly the level of MDA(9 9?1 8 vs. 5 5?0 4)?CK(122?24 vs. 49?11)?NO(270?98 vs. 128?46) in plasma and MPO(skeletal muscle 4 27?0 53 vs. 1 28?0 19, lung 2 61? 0 12 vs. 0 57?0 02) in tissues respectively( P

Based on traditional acupuncture theory and modern researches, the factors that cause the difference of acupoints synergy effect are summarized and analyzed. It is found that the factors include the specificity of acupoint, the interaction of acupoints, the pathway of acupuncture signal, the body condition level, acupuncture manipulation, etc. It is believed that the specificity of acupoint is the key factor to determine the difference of acupoints synergy effect. Interaction of acupoints may be related to the pathway of selected acupuncture signal, which is an important factor in difference of acupoints synergy effect. The body condition level and acupuncture manipulation are internal and external factor to influence acupoints synergy effect, respectively.
Acupuncture Points ; Acupuncture Therapy ; instrumentation ; methods ; Humans ; Meridians ; Signal Transduction ; Treatment Outcome

Objective To observe the initial therapy outcomes of Valproic acid for childhood absence epilepsy (CAE),and to assess its therapeutic reaction and short-term prognosis,and to investigate the risk factors for initial therapy failure.Methods From January 2010 to December 2015,absence seizures as key words were used to search CAE in the video-electroencephalogram(VEEG) database of Children's Hospital Affiliated to Nanjing Medical University.Sixty-seven children fulfilled the CAE diagnosis criteria of International League Against Epilepsy in 1989.These patients were separated into 2 groups based on the therapy outcome after 6 months,including seizure control failure group and seizure control group.The patients' clinical characteristics and VEEG characteristics were reviewed.The gender,age of seizure onset,a family history of epilepsy or febrile seizures (FS),consistent with 2005 Panayiotopoulos diagnostic criteria,and VEEG findings were analyzed to evaluate the predictive ability of independent variables and the relationship between these features and treatment outcomes by using a stepwise multivariate Logistic regression model.Results The age at seizure onset was (5.89 ± 2.91) years old,and follow up duration was 6 months.Approximately 23 cases (34.3％) of CAE patients had poor response.No statistical correlation was made for gender,age at onset,the occurrence of generalized tonic clonic seizures,and family history of FS or epilepsy between 2 groups (all P ＞0.05).Compared with seizure control group,patients of the seizure control failure group had significantly higher rates of focal epileptic discharge (87.0％ vs.6.8％),higher rates of intermittent photic stimulation (IPS) induced seizures (52.2％ vs.6.8 ％),fewer rates of occipital intermittent delta activity (8.7 ％ vs.77.3 ％),and fewer rates of patients met the new diagnostic criteria proposed by Panayiotopoulos in 2005 (8.7％ vs.88.6％) (all P ＜ 0.001).The presence of occipital intermittent rhythmic delta activity during wake stage and the interictal focal epileptiform discharges on VEEG during sleep stage were significantly associated with the therapy outcomes in a multivariable Logistic regression analysis (OR =133.714,P ＜ 0.05;OR =0.068,P ＜ 0.05).Conclusions The presence of focal epileptiform discharges,and a typical absence induced by IPS are important factors for first-drug treatment failures with CAE,and conversely the occipital intermittent rhythmic delta activities have a good efficacy.There is no statistical correlation between clinical characteristics and treatment outcomes.