Objective To investigate the effects of montelukast(MK),a selective cysteinyl leukotriene receptor 1 antagonist on interleukin(IL)-5 and eotaxin expression as well as serum total immunoglobulin E(IgE) production during MK treatment of mouse acute asthma airway inflammation.Methods Sensitized Balb/c mice were challenged by ovabumin(OVA)to establish the acute asthmatic mo-(del).Twenty-five mg/kg of MK(MK group) or Saline(Saline group)were given by intravenously for 3 days.Cellular infiltration of bronchoalveolar larvage fluid(BALF) were assessed by Wright staining.Production of IL-5 and eotaxin in the lung or BALF and serum IgE were determined by enzyme linked immunosorbent assay(ELISA).The expression of IL-5 and eotaxin mRNA were measured by semi-quantified reverse transcriptase-polymerase chain reaction(RT-PCR).Plasma MK level was determined by liquid chromatography.Results After 24 h OVA challenge,the numbers of total white blood cells,neutrophils and eosinophils(EOS)of BALF were(26.0?18.9)?10~7/L,(5.92?8.09)?10~7/L and(0.74?0.88)?10~7/L in MK treatment group.They were significantly reduced compared with those in Saline group,respectively(P80%.The level of IL-5 in BALF and lung tissue were(48.52?14.45) ng/L and(27.40?9.62) ng/g protein in MK group,which significantly declined compared with that in saline group;BALF eotaxin level declined too.Serum IL-5 and total IgE level were also significantly reduced;IL-5 mRNA expression in lung significantly decreased.Eotaxin and its mRNA expression in lung were not decreased significantly.Conclusion MK(exerts) its anti-inflammatory effect mainly through the suppression of IL-5 and IgE production.

Objective To observe the effect of Yiqixue Buganshen Recipes(YBR), the compound recipes of Chinese herbal medicine with the actions of replenishing Qi and blood and tonifying liver and kidney, on the endometrial integrin β3 of the mice with embryo implantation dysfunction, and to explore the molecular mechanism of YBR in improving embryo implantation. Methods Ninety mice were divided normal group, model group, and YBR group, 30 mice in each group. YBR group was given Jinghou Zengzhi granules 8mg/kg for promoting post-menstruation proliferation and Cuhuangti granules 8mg/kg for promoting the formation of corpora luteum. At 8∶00 am of pregnant day 4 (Pd4), subcutaneous injection of mifepristone was used for inducing embryo implantation dysfunction in the model group and YBR group. Twenty mice from each group were executed 12 hours after subcutaneous injection of mifepristone, and the uterus was isolated for the detection of mice endometrial integrin β3 mRNA and protein expression with real-time RT-PCR and Western blotting method respectively. Ten mice from each group at Pd4 were executed, and the uterus was isolated and then fixed in 2.5% glutaraldehyde solution for the detection of the formation of pinopode in mice endometrium under scanning electron microscope. Results The detection by real-time PCR and Western blotting method revealed that integrinβ3 mRNA and protein expression levels of the model group were significantly lower than those of the normal group (P<0.05), and YBR group had higher integrin β3 mRNA and protein expression levels than the model group(P<0.05). The formation of pinopodes in the model group was less than the normal group and YBR group, but the number of formed pinopodes in YBR group was similar to that in the normal group. Conclusion YBR can up-regulate the expression of integrin β3 in the endometrium of mice with embryo implantation dysfunction, and have positive regulatory effect on the formation of pinopodes on Pd4.

Objective To investigate the expression and distribution of plasmacytoid dendritic cells(pDC) in peripheral blood and renal tissues in children with Henoch-SchSnlein purpura(HSP),and explore the role of pDCs in the pathogenesis of Henoch-Schtnlein purpura nephritis(HSPN).Methods Among the 40 children with HSP,28 cases were in the active phase(renal biopsy performed in 8 cases of them) and the other 12 in remission phase.Peripheral blood mononuclear cells were isolated,and the expression of pDC was detected by flow cytometry.The normal control group was established (n =15).Total RNA of peripheral blood was extracted and transcripted into cDNA.Sybr green dye based real-time quantitative PCR method was used to compare the expression(indicated as 2-△Ct value) of CXC motif chemokine 10 (CXCL10),CC chemokine ligand 5 (CCL5),chemokine CXC subfamily receptor 3 (CXCR3),CC chemokine receptor 5 (CCR5) in children with HSP and those in the controls.Immunohistochemistry labeling technique was used to detect the distribution of pDC in renal tissues from renal biopsy,and the normal controls were established (n =3).Results The expression percentage of pDC in peripheral blood in active phase was 0.051 ± 0.039,significantly lower than those in remission phase (0.181 ± 0.082) and the normal controls (0.166 ± 0.079) (P ＜ 0.000 1).Chemokines genes CXCL10 and CCL5 were overexpressed in peripheral blood ceils of acute phase HSP children,but chemokine receptors CXCR3,CCR5 were lowly expressed compared with normal controls.There was almost no expression of pDC in the normal control renal tissues,while pDC was infiltrated in glomeruli of HSPN children.Conclusions The number of pDC and chemokines' expression in peripheral blood is abnormal,and the pathogenesis of nephritis may be involved with the pDC in peripheral blood to migrate to the renal tissues.

Objective To localize the tibial attachment of the posterior cruciate ligament (PCL) on the magnetic resonance imaging (MRI) and provide parameters for clinical PCL reconstruction.Methods We retrospectively analyzed 524 patients with intact tibial PCL attachment who had undergone knee MRI from January 2010 to January 2016.They were 286 men and 238 women with an average age of 35 years (from 20 to 50 years).The size and positions of the tibial PCL attachment were measured on the sagittal and coronal MRI slices.The differences were analyzed between different genders.Results On the sagittal slices,the mean distance from the central tibial PCL attachment to the posterior edge of the tibial plateau was 17.9 ± 3.0 mm and the mean anteroposterior diameter of the tibial PCL attachment was 9.7 ± 2.4 mm,with those for males significantly larger than for females (P ＜ 0.05).The above mean values when expressed as a percentage of the posterior tibial slop were 79.9％ ±4.5％ and 43.7％ ± 9.6％,respectively,showing no significant differences between males and females (P ＞ 0.05).On the coronal slices,the distances from the central tibial PCL attachment to the medial and lateral edges of the tibial plateau were 33.5 ± 3.1 mm and 37.4 ±4.1 mm,respectively,and the mediolateral diameter of the tibial PCL attachment was 12.0 ± 1.6 mm,with those for males significantly larger than for females (P ＜ 0.05).The above mean values when expressed as a percentage of the mediolateral diameter of the tibial PCL attachment were 47.4％ ± 3.2％,52.7％ ±3.1％ and 16.9％ ± 1.7％,respectively,showing no significant differences between males and females (P ＞ 0.05).Conclusions On knee MRI images,the distance from the central tibial PCL attachment to the posterior edge of the tibial plateau is about 17.9 mm,the anteroposterior diameter of the tibial PCL attachment around 9.7 mm,and the mediolateral diameter of the tibial PCL attachment roughly 12.0 mm.These measurements for males are larger than for females.

Objective To investigate the expression of survivin in T lymphocytes that were stimulated by Con A and alloantigens in renal grafts in vitro and in vivo. Methods According to the different treatments, the experiment was divided into three parts. (1) The C57BL/6 mice splenocytes stimulated by Con A (10 mg/L) were cultured in the RPMI-1640 medium. Following the proliferation blockade or not, the expression of Survivin in the splenocytes was detected. (2) The GVHR model was established by transfusing the C57BL/6 mice splenocytes into Balb/c× C57BL/6 F1 mice, and the expression of Survivin in the donor splenocytes was detected at the different time points. (3) Seventythree cases of clinical renal allograft biopsy specimens were collected, pathologically diagnosed and classified according to the Banff 97 classification, and then the expression of Survivin was detected.Results Survivin was expressed in the CD3+ splenocytes that received Con A stimulation. The positive cell count reached the peak on the day 3, and subsequently declined. In the GVHR model, the lymphocytes infiltration and Survivin expression were detected around the portal vein and portal area on the post-splenocytes-transfused day (PSTD) 4 to 12. But on the PSTD 14, the Survivin expression could not be detected in the infiltrated lymphocytes. In the renal allograft biopsy specimens,lymphocytes did not express Survivin in 13 specimens of the group without acute cellular rejection.was difference between the two groups (P＜0. 01 ). Conclusion The activated T cells possess the capacity to express Survivin, and the expression is time-dependent. For the characteristics of Survivin expression of T cells, it may be applied as an approach to diagnose the acute cellular rejection and judge its degree and stage in the clinical allograft biopsy specimens.

Objective To develop the hypothesis ‘saturated or non-saturated cytotoxicity model' and explain the various phenomena of antibody mediated immunoresponses in recipients,including rejection and accommodation.Methods The imitating complement dependent cytotoxicity.The threshold set to identify as saturated or non-saturated cytotoxicity depends on antigen-antibody complex(R)whether or not above lethal number(D)in effective time.Feasibility of the hypothesis was examined through explaining various phenomena mediated by anti-donor antibodies,especially some contradictory phenomena.Results Hyperacute rejection,accelerated rejection and acute rejection could be well explained by saturated cytotoxicity.Accommodation of ABO imcompatible transplantion,de novo antibody induced injury,change of protein profile,and C4d deposition in graft could be well elucidated by the hypothesis.Conclusion The hypothesis saturated or nonsaturated cytotoxicity model' help to interpret and interconnect various phenomena of antibodies mediated immune response,such as rejection and accommodation.

A fragment sized 400bp of White spot syndrome virus（WS SV，formerly de signated NOSV），recovered from recombinant plasmid pAFD, was labeled with Digox igenin as a probe to detect dynamic distribution of WSSV within 120h and 72h in crawfishes（Cambarus proclarkii） inoculated WSSV by oral taking and injecti on r espectively. Stomach epithelium, intestine epithelium, heart, gill, haemolymph, muscle, hepatopancreas, hypoderm, connective tissue and ovary of infected crawfi shes were examined for WSSV. In both groups, WSSV was first detected in heamoly mph at 12h p.i. and then disappeared. Again it was detected at 96h p.i. only in oral infection group and maintained till 120h p.i., but it didn't appear at 72h p.i. in injection group. WSSV in heart, muscle was detected at 36h p.i. in oral infection group and 24h p.i. in injection group respectively, and then increased generally. In addition, WSSV in intestine epithelium, connective tissue, ovary of oral infection group and intestine epithelium, hypoderm, ovary of injection g roup could also be detected. In dead crawfishes after 120h and 72h p.i. in two groups, WSSV could be detected in all the examined tissues and it demonstrated t hat systemic infection occurred in the animales. The tissue containing more amo unts of WSSV was hypoderm in oral infection group, while intestine epithelium, g ill, hypoderm, ovary in injection infection group. It deduced that WSSV first a ppears in haemolymph and then goes into heart, muscle and other tissues and prol iferates in them. Once again, WSSV is released into heamolymph resulting in syst emic infection till crawfishes' death.

OBJECTIVETo investigate the expression of PCNA and p27 in human benign prostate hypertrophy (BPH) and prostate carcinoma (PCa) and their effect on the genesis and progression of the tumor.

METHODSThe paraffin-embedded sections of 30 cases with BPH and 37 cases with PCa were collected. The expression of p27 and PCNA protein were examined by S-P immunohistochemical method. Comparative analysis for BPH and pathological grade and clinical stage of PCa was performed.

RESULTSThe expression of PCNA in BPH (3.3%) was significantly lower than that in Pca (83.8%, P < 0.01). The expression of p27 in BPH (70.0%) was significantly higher than that in Pca (27.0%, P < 0.05). The expression of p27 was not correlated with histological grade and clinical stage in Pca (P > 0.05). An inverse correlation was found between p27 and PCNA expression in BPH (P < 0.01), while no correlation was found in Pca (P > 0.05).

CONCLUSIONThe loss or decreased expression of p27 protein may be related to the genesis of benign prostate hypertrophy, but not to the development of prostate carcinoma; the overexpression of PCNA may play an important role in the malignant behavior and progression of prostate carcinoma.

Adenocarcinoma ; metabolism ; pathology ; Aged ; Aged, 80 and over ; Cell Cycle Proteins ; metabolism ; Cyclin-Dependent Kinase Inhibitor p27 ; Gene Expression Regulation, Neoplastic ; Genes, Tumor Suppressor ; Humans ; Male ; Middle Aged ; Neoplasm Staging ; Prognosis ; Proliferating Cell Nuclear Antigen ; metabolism ; Prostatic Hyperplasia ; metabolism ; pathology ; Prostatic Neoplasms ; metabolism ; pathology ; Tumor Suppressor Proteins ; metabolism

OBJECTIVETo establish enzyme-linked immunosorbent assay (ELISA) for quantitative detection of Golgi protein73 (GP73) in serum.

METHODSA sandwich reaction was preformed with horseradish peroxidase labeled monoclonal antibody of GP73 as the catalytic enzyme. Several reactions liquid's concentration and reaction conditions were optimized. The method was evaluated in all aspects such as linear range, sensitivity, specificity, stability and so on.

RESULTSThe linear range was 25-500 ng/ml. The detection limit was 18.5 ng/ml. Inter-assay and intra-assay RSD were both less than 10%. The recoveries of three different spiked concentration samples were 95.3%, 92.6% and 103.7%. After stored at 4 degrees C and 37 degrees C for 3, 5, 7 days, the analysis showed correlation coefficient higher than 0.98 and RSD lower than 10%.

CONCLUSIONEstablished ELISA for quantity determination of serum GP73 has high accuracy, sensitivity and repeatability.

Antibodies, Monoclonal ; analysis ; Enzyme-Linked Immunosorbent Assay ; methods ; Humans ; Membrane Proteins ; blood ; Sensitivity and Specificity

Objective To investigate the association of lipoprotein lipase gene Hind Ⅲ and S447X polymorphisms with metabolic syndrome among Kazakh and Han ethnicities in Xinjiang.Methods PCR-RFLP was used to detect 802 subjects' lipoprotein lipase Hind Ⅲ and S447Xgenotypes (including 201 controls and 200 metabolic syndrome patients in Kazakh and Han ethnicities, respectively). Results (1) Frequencies of H + H-/H-H- genotype (32.50% vs.47.76%), H- allele( 18.00% vs. 28.86%), SX/XX genotype (8.00% vs. 22.39%) and X allele (4.00%vs. 12.44% ) for metabolic syndrome in Hah ethnicity were all significantly lower than those in controls (P＜ 0.01 ). (2) The frequencies of H + H-/H-H- genotype (33.50% vs. 46.80% ), H- allele (22.00% vs. 28.60%), SX/XX genotype (10.50% vs. 22.90%) and X allele (5.50% vs. 12.44% ) in patients with metabolic syndrome in Kazakh were all significantly lower than those for controls (P＜0.01). (3) The frequencies of lipoprotein lipase gene Hind Ⅲ and S447X genotypes and alleles in Kazakh were not significantly different from Han (all P＞0.05). (4)The levels of waist circumference, systolic blood pressure, diastolic blood pressure, triglyceride and FPG in H + H-/H-H- and SX/XX genotype were significantly lower than those in H + H + and SS genotype.HDL-C was significantly higher than that in H + H + and SS genotype (P＜0.05). (5) The frequencies of H + H + and SS genotype increased along with the increase in number of metabolic syndrome component. Conclusion The lipoprotein lipase gene Hind Ⅲ and S447X polymorphisms were associated with metabolic syndrome risk in Kazakh, and H + H-/H-H- genotype, H- allele, SX/XX genotype and X allele might have served as protective factors of metabolic syndrome. H + H-/H-H- and SX/XX genotype seemed to have had beneficial effects for all the metabolic syndrome components, and the frequencies of H + H + and SS genotype were increasing along with the increase of number in the metabolic syndrome components.