Objective:To investigate the mechanism of immunoglobulin Y antibodies(IgY) against tumour necrosis factor alpha(TNF-α) and Interleukin-1beta(IL-1β) in treating allergic bronchial asthma through nebulization inhalation.Methods:The allergic bronchial asthma model was established with Hartley guinea pig by ovalbumin nebulizating inhalation.The animals were randomly divided into 4 groups: normal control group(group A)，allergic bronchial asthma group(group B),0.1% anti-TNF-α and IL-1β IgY treating group(group C),1.0% anti-TNF-α and IL-1β IgY treating group (group D).The animals were killed after treatment being accomplished for 2 h,4 h,8 h,24 h and the lungs were made pathological,which were then stained by hematoxylin-eosin(H.E.).The bronchoalveolar lavage fluid (BALF) was collected and the deposited cells were stained by Wright's.Results:①The histological appearance of lung: In group B the histological structure of alveolar ducts and alveolar walls was damaged,the alveolar space was full of transudate and lots of alveolar epithelial cells and leucocytes.The pulmonary interstitial edema,inflammatory cells infiltration,distorted or dilated capillaries and reducing capillary numbers of effective blood stream were observed in alveolar walls.In group C and group D the damage degree of alveolar ducts and alveolar wall was slighter than that in group B and there were few inflammatory cells in alveolar space.In bronchial lumen and pulmonary alveoli the sticky mucus plug was obviously less in group C and group D than in group B.Moreover,inflammatory cell infiltration was seldom observed aroud bronchia,and restoration of bronchial tunica mucosa epithelium was obviously observed in group C and group D.②The cytology appearance of BALF: In group C and group D the numbers of eosinophils,neutrophils,lymphocytes were significantly fewer （2 h,4 h,8 h,P＜0.05）,however,the number of macrophage was significantly more（2 h,4 h,8 h,P＜0.05）than in group B.Conclusion:The anti-TNF-α and IL-1β IgY can obviously alleviate pathological extent of inflammatory reaction in allergic bronchial asthma of guinea pigs by nebulization inhalation therapy.The therapeutic effect of anti-TNF-α and IL-1β IgY between 0.1% and 1.0% concentration is not obvious difference for pathology changes.

AIM: To evaluate the relationship between axial length ( AL ) , corneal and lens refractive power, and the refractive error in children.
METHODS:Totally 44 children 88 eyes with refractive error who underwent retinoscopy with cycloplegia, to be measured spherical equivalent refractive error. Axial length was measured by a noncontact optical biometry ( ZEISS IOL-Master) , and corneal K and anterior chamber depth ( ACD) were also measured by the same machine. The refractive power of the lens was calculated by using the SRK formula. The patients were divided into 3 groups, myopia (SE<0), hyperopia (SE>+0. 50D) and emmetropia ( 0 to + 0. 50D ). Linear Correlation and Regression were used to evaluate the correlation among the optical parameters.
RESULTS:Totally 44 subjects, 88 eyes, average 9. 04±2.39 years, spherical equivalent (SE) -3. 50D to +8. 75D. Hyperopic AL was shorter than the other two groups ( P<0.05), same trend in lens power. No significant differences among the 3 groups in corneal K and ACD. There was a negative correlation between age and SE, SE and AL, SE and lens refractive power, and there was a positive correlation between age and AL, age and lens power.
CONCLUSION:As the children's growing up, SE trends to myopia, and AL becomes longer, and lens power is stronger.

Objective To further analyze the chaos characteristics of heart rate variability(HRV) signals.Methods Analyzed HRV was obtained from 40 healthy subjects(20 olds,20 youngs) by means of Cao algorithm;Stability criteria was proposed to determine minimum embedding dimension;Meanwhile,we advanced a new quantative analysis method of studying HRV deterministic component,and applied the method to the old and young groups.Results Each subject's HRV was composed of both stochastic and deterministic components;Stability criteria could solve the subjective problem when determine minimum embedding dimension by Cao algorithm;Deterministic component in healthy old subjects was higher than the young.Conclusion In this paper,we have proved that HRV is deterministice chaos;Stability criteria works well for determining minimum embedding dimension;HRV decreases as age increases.

Objective To investigate the clinical characteristics and classification of ketosis-prone diabetes. Methods Firstly,according to with or without autoantibodies,patients with an initial onset of unprovoked ketosis or ketoacidosis were classified to two different groups. Secondly,based on weather or not depending on insulin,patients without autoantibodies were further divided into two groups,insulin dependent and non-insulin dependent. Clinical characteristics,biochemical parameters were compared between different groups. Results The autoantibodies negative group showed a strong male predominance and family history of diabetes.BMI was higher than type 1 diabetes. Plasma triglyceride level was highest. Fasting and postprandial C-Peptide level was between the other two group. Compared with non-insulin dependent group,insulin dependent group showed a strong male predominance,lower ratio of overweight and obesity,more severe ketosis status. There were no significant differences with regard to blood glucose level and fasting C-Peptide level of initial. But fasting C-Peptide after 3 months was higher in non-insulin dependent group . Conclusion Ketosis-prone diabetes can divided into two groups,based on autoantibody negative or positive. The latter consist of type 1A diabetes mellitus. The former can further classified as ketosis-prone type 2 diabetes and idiopathic type 1 diabetes(type 1B diabetes)according to insulin dependent or non-insulin dependent.

The incidence of birth defect in China is still high,which is not only affected by genetic factors,but also affected by heavy metals in surrounding that prejudices foetus' normal development.Heavy metal is a kind of inorganic pollutants with high toxicity.Excessive intake of lead,cadmium,arsenic or mercury,and the insufficient intake of zinc,calcium and copper may both lead to at least 1 kind of birth defect.The interactions between heavy metals also affect the outcomes of pregnancy.This paper reviewed different relationships between heavy metals and birth defect recording to relevant achievements such as animal experimentations and epidemiologic study made by researches from at home and abroad in recent years.It is advised to intake adequate zinc and copper and avoid being exposed to harmful metals to make sure the effective reduction of the incidences of birth defects.This paper will also point out the direction of future research about the relationship among the birth defects and heavy metal.

Objective To evaluate the accuracy of stoke volume variation (SVV) determined using FloTrac/Vigileo and Picco-plus technologies in prone position for assessment of the blood volume in the patients undergoing spine surgery,Methods Forty-three ASA physical status Ⅰ-Ⅲ patients of both sexes,aged ＞ 18 yr,weighing 40-100 kg,scheduled for elective posterior approach to lumbar spinal fusion or scoliosis surgery were studied.After induction of anesthesia,a volume expansion was performed in supine and prone positions.Hydroxyethyl starch 130/0.4 sodium chloride injection 5 ml/kg was rapidly infused intravenously over 10 min to carry out the test for fluid responsiveness.Picco-plus and FloTrac/Vigileo systems were simultaneously applied in every subject to measure SVV (SVVP and SVVF).Positive fluid responsiveness was defined as the changing rate of stroke volume index ≥ 10％ as measured by using Piccoplus system.The patients were divided into response group (Rs group) and non-response group (NRs group) according to the changing rate of stroke volume index ≥ 10％ and ＜ 10％.The receiver operating characteristic (ROC) curve for SVV was plotted,and the diagnostic threshold,area under the ROC curve and 95％ confidence interval (CI) were calculated.Results Forty-one patients were included for analysis in this study.In supine position,the area under the ROC curve for SVV in predicting the fluid responsiveness was 0.740 (95％ CI:0.568-0.913),the diagnostic threshold was 12％,and the sensitivity and specificity in determining fluid responsiveness were 86％ and 54％,respectively,for SVVF,and the area under the ROC curve was 0.637 for SVVP.In prone position,the area under the ROC curve was 0.451 for SVVF,and 0.634 for SVVP.Compared with Rs group,the baseline value of SVVFwas significantly lower,and no significant change was found in the other hemodynamic parameters before volume expansion in supine position in NRs group.There was no significant difference in the hemodynamic parameters before volume expansion in prone position between the two groups.Conclusion SVV determined by using FloTrac/Vigileo and Picco-plus systems in prone position can not accurately assess the blood volume in the patients undergoing spine surgery.

Objective To clone NT-3 gene from normal rat brain and to purify its fusion protein and to prepare specific high titer antibody so that to provide a foundation for further study for peripheral nerve injury.MethodsWe amplified target gene by RT-PCR and cloned it into the vector of pMD-18T,then analyzed its sequence and compared it with the sequence from GenBank.We subcloned it into pRSET-A vector and introduced it into Escherichia coli BL21.The expression was induced by IPTG,and identified by SDS-PAGE.The fusion protein was purified by niccolum purify kit.We immuned rabbits with immunological adjuvant for specificity antibody preparation.Results We got a 777 bp gene segment by RT-PCR.The DNA sequence was identical to rat NT-3 gene sequence in GenBank.It proved that the target gene was correctly inserted into the vector.A new protein band of about 34 ku appeared on SDS-PAGE after induction of IPTG.A specific high titer antibody of 1∶64000 was gained by immunizing rabbits with adjuvant.

AIM: To study the expression of hepatitis B virus core gene in Pichia pastoris and to obtain high-level expressed recombinant HBcAg with good immunoreactivity and high specificity. METHODS: HBV core gene was amplified by PCR from plasmid pHBV1 which contained HBV whole DNA sequence. The PCR product was cloned into pGEM-T vector by TA cloning strategy. After confirmed by DNA sequence analysis, the gene of interest was inserted into the yeast expression vector pPIC9. The recombinant plasmid pPIC9-cAg was constructed and transformed into GS115 by electroporation. The recombinant yeast GS115 was induced by 0.5% methanol. The expressed product was analysed by SDS-PAGE,Western blot and ELISA. RESULTS: The restriction analysis and DNA sequence analysis proved that HBV core gene had already been cloned to yeast expression plasmid pPIC9. The expressed HBcAg existed in SDS-PAGE. Good immunoreactivity and high specificity of the recombinant HBcAg have been proved by ELISA and Western blot. The titre of the recombinant HBcAg in the cell lysate was 1∶ 12 800 . CONCLUSION: The recombinant plasmid pPIC9-cAg was successfully constructed. The recombinant HBcAg with good immunoreactivity and high specificity was successfully expressed in Pichia pastoris expression system and can be applied to further developing HBcAb immunoassay. [

Objective:To investigate the effects of epidermal growth factor (EGF)on cell cycle and cell cycle-related regulatory factors of human esophageal squamous cell carcinoma (ESCC) cell line Eca109.Methods: Serum starved Eca109 cells were treated with 20 ng/ml recombinant human EGF(rhEGF)for 24 h.The cell cycle phase distribution was detected by flow cytometry.The mRNA and protein expression levels of p21CIP1/WAF1(p21) and p27KIP1(p27) were detected by real-time quantitative reverse transcription polymerase chain reaction(qRT-PCR)and Western blot,respectively.Results: The proportions of G1 phase cells in EGF group and control group were ( 54.90 ±0.82 )% and ( 65.94 ±0.74 )%.The mRNA and protein expression levels of p 21 in EGF group was significantly higher ,and p27 was significantly lower than that in control group ( P<0.01 ) .Conclusion: EGF facilitates G1-S phase transition,and promotes the proliferation of Eca 109 cells,which may be associated with the up-regulation of p21 and down-regulation of p27.