Objective:To construct the expression vectors of VH and VL from an anti -CD71 monoclonal antibody (line7579) , and express an anti-CD71 mouse/human chimeric antibody (D2C) in vitro.Methods:The variable regions of heavy and light chains from an anti -CD71 monoclonal antibody (line7579) cloned in the pGEM-T vectors were constructed respectively into the expression vectors,p?-Expr(including human IgG1 constant region) and p?-Expr(including human Ig? constant region).The expression plasmid vectors (Chi7579-p?-Expr and Chi7579- p?-Expr) ,confirmed by a clone-PCR and a further sequence analysis, were co-transfected by electroporation into the non-Ig-producing murine hybridomas SP2/0.Results:Screening for transfectomas secreting chimeric antibody was done by cross sandwich ELISA, which identified the transfectomas to secrete a chimeric antibody (D2C) with the human heavy constant region(C?) and light constant region(C?). FCM analysis of culture supernatants of the transfectomas demonstrated that the binding capacity of the chimeric Ab (D2C) to the antigen (CD71) was retained.Conclusion:The construction of the V H and V L expression vectors and the expression of an anti-CD71 mouse/human chimeric antibody (D2C) were accomplished successfully.

Aim To observe the protective effects of breviscapine against lung fibrosis and investigate its possible mechanism.Methods Effects of breviscapine on cell proliferation,activation and extracellular matrix secretion were examined in mouse embryonic lung fibroblast L929 cells in vitro.The mouse model of bleomycin-induced lung fibrosis was used to assess the protective effect of breviscapine against lung fibrosis.Results In vitro,breviscapine had no cytotoxicity directly on L929 cells,however,it could suppress cell proliferation,activation and secretion of laminin(LN) and collagen Ⅰ(ColⅠ) induced by transforming growth factor beta1(TGF-?1) in L929 cells.In vivo,breviscapine could prevent increase in serum TGF-?1 and decrease in superoxide dismutase(SOD),peroxidase(POD) and catalase(CAT) in mice with lung fibrosis caused by bleomycin.In addition,breviscapine was able to reduce pulmonary hydroxyproline,collagen,malondialdehyde(MDA)and TGF-?1 in lung fibrosis mice.Conclusion Breviscapine has protective effect against lung fibrosis and the possible mechanism is to enhance antioxidative defense activities and prevent TGF-? signal.

Objective To develop a dental crown remover with the strength provided constantly and adjusted freely.Methods The remover was composed of a shell, a crown hook, a conduction mechanism, a strength generating mechanism and a percussion mechanism. The spring had the explosive force formed in the crown head to destruct the cohesion of the cement.Results The remover could remove the crown, with one hand freed for the protection of abutment and mucous membrane.Conclusion The remover has simple structure and easy operation, and is worth popularizing clinically.

Adult ; Aged ; Female ; Humans ; Lung Neoplasms ; blood ; genetics ; Male ; Middle Aged ; Mutation ; Tumor Suppressor Protein p53 ; blood ; genetics

Objective To construct the pshuttle-Egr-1-hSmac plasmid and transfect human breast cancer MDA-MB-435 cells,and to observe its radiotherapy enhancing effect on tumor cells.Methods The empty vector pshuttle and pshuttle-Egr-1-hSmac plasmid were transfected into MDA-MB-435 cells by liposomal.At different time(4,8,12,24 and 48 h)after irradiation with 2.0 Gy X-ray and 24 h after irradiation with 0.5 -5.0 Gy,the total RNA and protein were collected and extracted from these cells to analyze the Smac mRNA and protein expression levels with RT-PCR and Western blotting methods. The cells were divided into control, pshuttle, pshuttle-Egr-1-hSmac,2.0 Gy irradiation group, pshuttle + 2.0 Gy irradiation and pshuttle-Egr-1-hSmac+2.0 Gy irradiation groups.MTT method was used to evaluate cell proliferation,and the cell survival ability was measured with clone formation assay;Annexin Ⅴ/PI double staining and PI single staining were used to examine the apoptosis and cell cycle of MDA-MB-435 cells. Results There was no Smac mRNA expression in MDA-MB-435 cells in control and pshuttle groups,but the Smac mRNA expression levels in MDA-MB-435 cells in pshuttle-Egr-1-hSmac plasmid group were gradually increased with the time prolongation, and reached the maximum at 24 and 48 h;the Smac mRNA expression levels in MDA-MB-435 cells were increased gradually 24 h after irradiation of 0.5 - 5.0 Gy X-ray with the increasing of irradiation doses, and reached the maximum after 2.0 and 5.0 Gy irradiation. The Smac protein expression levels in pshuttle-Egr-1-hSmac plasmid group were increased gradually with the time prolongation,and reached the maximum at 24 h.The Smac protein expression lervels were increased 24 h afer irradiation of 0,0.5,1.0,2.0 and 5.0 Gy X-ray,especially in 5.0 Gy group. The MTT results showed that the A490 values in 2.0 Gy,pshuttle+2.0 Gy and pshuttle-Egr-1-hSmac groups 24, 48,and 72 h after irradiation were lower than those in control group(P<0.01);the A490 values of MDA-MB-435 cells in pshuttle-Egr-1-hSmac group after 1.0-5.0 Gy X-ray irradiation were lower than those in 0 Gy group (P<0.05 or P<0.01);the survival fraction(SF)in pshuttle-Egr-1-hSmac group was lower than those in control group (P<0.01).The percentages of the cells at G0/G1 and S phase in pshuttle-Egr-1-hSmac group were lower than those in 2.0 Gy group(P<0.01),the percentage of the cells at G2/M phase was higher than that in 2.0 Gy group (P<0.01);the apoptotic rate of the cells in pshuttle-Egr-1-hSmac group was higher than that in 2.0 Gy group (P<0.01).Conclusion X-ray irradiation can significantly increase the Smac mRNA and protein expression levels in MDA-MB-435 cells transfected with pshuttle-Egr-1-hSmac plasmid,inhibit the cell survival rate,and induce G2/M arrest and apoptotic increasing;Smac gene combined with radiotherapy could significantly increase the radiosensitivity of breast cancer cells.

Objective Glucose metabolism trend was dynamicly mornitored following liver transplantation, and its affecting factors were assessed. Methods The glucose metabolism status were assessed at four time points respectively after liver transplants, then they were divided into two groups:normal glucose metabolism (NGM) and abnormal glucose metabolism (AGM). The clinical data were univariate analyzed and multivariate analyzed to screen the risk factors. Results At 1 month, 3 months, 1 year and 3 years post-transplantation, the incidence of AGM were 74.0%, 43.9%, 29.4%, 24.1% respectively Between these two groups, age > 45 y had a significant difference at 1 month, 3 months, 1 year and 3years post-transplantation; the use of tacrolimus had a significant difference at 3 months, 1 year and 3years post-transplantation, but the dose of tacrolimus or tacrolimus blood concentration showed no significant difference; high dose of glucocorticoid had significant difference at 1 month , 3 months post-transplantation; high BMI and acute rejection had significant difference at 1 month post-transplantation. Conclusions There is a high incidence of abnormal glucose metabolism (AGM) in the early stage post-transplantation, and a considerable number of patients' glucose metabolism improved in the later period. Age>45 y and tacrolimus affect glucose metabolism for a longer period post-transplants. High BMI and acute rejection have an impact on glucose metabolism only in the early stage post-transplantation. Large dose of glucocorticoid affect glucose metabolism for at least 3 months post-transplantation , and there is no significant difference after 1 year.

Objective:To study the relationship between neutrophil to lymphocyte ratio (N/L) and traditional cardio‐vascular risk factors among community 35~64‐year‐old residents .Methods :A total of 1884 residents (548 males and 1336 females) from urban Shenyang city received baseline condition questionnaire on cardiovascular diseases and re‐lated diseases from Apr 2011 to Feb 2012. According to presence of cardiovascular risk factors or not ,subjects were divided into healthy control group (n=675) and risk factor group (n=1209);according to number of risk factors , risk factor group was further divided into one risk factor group (n=491) ,two risk factors group (n=263) and ≥3 risk factors group (n=455) .Morning blood sample and urine sample were retained to measure blood and urine rou‐tine ,blood glucose and blood lipid profile etc in all subjects .N/L was compared and analyzed among all groups .Re‐sults:Among patients with only one of following risk factors [hypertension ,diabetes mellitus (DM) ,dyslipidemia and obesity] ,N/L levels of patients with hypertension or DM were significantly higher than that of healthy control group [1.55(1.15 ,1.95) ,1.60(1.21 ,2.07) vs .1.45(1.09 ,1.91)] , P<0.05 both ,and there were no significant difference between any other one risk factor group and healthy control group , P>0.05 all .Among risk factor sub‐groups ,N/L level of ≥3 risk factors group was significantly higher than that of two risk factors group [1.57(1.16 , 2.04) vs .1.41(1.07 ,1.89) ,P<0.05] ,and there was no significant difference between any other two groups (P>0.05) .Conclusion:N/L significantly related to hypertension or DM ,and N/L level of ≥3 risk factors group was sig‐nificantly higher than that of two risk factors group ,N/L is helpful to assess risk of cardiovascular diseases .

Toxicological pathology is a morphological elucidatet ion of tissue damage caused by drug toxicity,including damage type,location,severity grade and prognosis.It provides objective and accurate data to support preclinical drug safety evaluation.Therefore,toxicological pathology plays an important role in drug safety evaluation,and the pathologic data or conclusion often determines the termination or continuation in many drug development projects.In this paper,the characteristic of toxicological pathology,standardized management,the development of new techniques including in situ hybridization,laser scanning cytometry and laser capture microdissection,and related problems in pathology are discussed briefly.

ObjectiveTo explore the prevalence and spectrum of β-thalassemia mutations in Fujian province,and to provide a reference for prenatal diagnosis and genetic counseling in this population.Methods Two thousand three hundred and one blood samples were randomly selected from 9 different areas of Fujian province from May 2008 to December 2010.PCR and reverse dot blot hybridization (RDB) were adopted for detection of the 17 common types of mutation,and the frequency of each genotype of β-thalassemia mutations was calculated.The β-globin gene of unknown positive samples were analyzed directly with DNA sequencing.Results Three hundred and fifty-nine cases were detected with β-thalassemia mutations of the 2301 copy blood samples submitted,and the detection rate was 15.60％ (359/2301).Of the mutated genes,12 different mutations were identified,namely IVS-2-654(C→T),CD41-42(-TCTT),CD17(A→T),-28(A→G),CD27-28(+C),CD26(G→A),CD71-72(+A),IVS-1-1(G→T),CD43(G→T),-29(A→G),initiation codon ATG→AGG and CD36(-C).Mutation frequencies were 46.54％ (175/376),33.24％ (125/376),9.31％ (35/376),5.05％ (19/376),2.13％(8/376),1.33％(5/376),0.80％(3/376),0.27％(1/376),0.27％(1/376),0.27％(1/376),0.53％(2/376),and 0.27％(1/376),respectively.The most common mutations were IVS-2-654 (C→T) and CD41-42 (-TCTT),which accounted for 79.78％(300/376) of total genetic mutations.In addition,a novel β-globin gene mutation CD36 (-C) allele was detected for the first time,the deletion of a nucleotide C at code 36 within exon 2 lead to a frameshift mutation that could result in a premature termination at code 60.Conclusions β-thalassemia mutations in Fujian province are complex with significant genetic heterogeneity.We present for the first time the detection of a new β-thalassemia mutation in the population:CD36(-C),which provides valuable information for genetic counseling and prenatal diagnosis in Fujian province.

OBJECTIVETo compare the efficacy and safety of a 6-week cluster schedule of specific immunotherapy with that of a 14-week conventional schedule for the treatment of subjects with persistent allergic rhinitis (AR).

METHODSThe trial was a prospective and randomized study involving 80 patients with persistent AR, who were allergic to Dermatophagoides pteronyssinus. While 40 patients were randomly assigned to the cluster schedule reaching the maintenance dose within 6 weeks, the other 40 patients were randomly assigned to the conventional schedule reaching the maintenance dose within 14 weeks. Symptom scores and medication scores were used to evaluate the clinical efficacy. Serum specific IgG4 level was examined to mark immunologic change, adverse reactions were recorded during the treatment.

RESULTSCluster group achieved clinical efficacy (reducing symptom scores and medication score) and increasing serum specific IgG4 sooner (after 6 weeks treatment). During the incremental dose phase, there were 6 systemic adverse reactions (1.12% of all injection) in 3 patients in cluster group and there were 5 systemic adverse reactions (0.85% of all injection) in 3 patients in conventional group. No severe systemic reactions occurred in both group. There was no difference between the 2 groups in frequency or type of systemic reaction (χ(2) = 0.333, P > 0.05).

CONCLUSIONThe cluster schedule is a safe alternative to the conventional schedule with the advantage of achieving clinical efficacy sooner.

Adolescent ; Adult ; Allergens ; immunology ; Animals ; Dermatophagoides pteronyssinus ; immunology ; Drug Administration Schedule ; Female ; Humans ; Immunoglobulin G ; blood ; Immunotherapy ; methods ; Male ; Middle Aged ; Prospective Studies ; Rhinitis, Allergic, Perennial ; therapy ; Young Adult