Objective:To identify the function of extracellular soluble vimentin that promotes proliferation, activation and chemotaxis of inflammatory cells. Methods: The proliferation of rat splenocytes stimulated with vimentin was evaluated in vitro. The lymphocyte counts and vimentin-antibody levels of peripheral blood in mice immunized with vimentin were detected in vivo. Peritoneal macrophages were collected and cultured with different concentrations of vimentin to detect the effect on phagocytosing chicken red blood cells. The chemotaxis of NIH3T3 fibroblast towards vimentin was observed in transwell chamber. Results:In vitro vimentin dose dependently promoted the proliferation of splenocytes. The proliferation indexes of primed and naive splenocytes cultured with 16μg/ml vimentin were reached up to ( 196. 0 ± 9. 7 )% and ( 208. 9 ± 4. 6 )% respectively without significant difference. In vivo vimentin significantly enhanced the lymphocytes number(109 L-1)of peripheral blood(5. 74±0. 51 vs. 1. 69±0. 13)and the levels of vimentin specific antibody( OD value 2. 31 ± 0. 06 vs. 0. 19 ± 0. 08 ) that shown no significant difference from immunization with vimentin plus CFA. In vitro vimentin dose dependently stimulated phagocytic ability of macrophages and performed the chemotactic effects on NIH3T3 fibroblasts. Conclusion:Extracellular soluble vimentin promotes the proliferation,activation and chemotaxis of concerned inflammatory cells and possesses the functions as an alarmin.

Objective To evaluate the treatment of PCI on hibernating myocardium..in ischemic cardiomyopathy with heart failure.Methods All patients identified iscbemic cardiomyopathy with heart failure by coronary angiography were randomly divided into PCI group and contrast group.61 patients in contrast group were just received the treatment of medi- cine while 60 patients in PCI group were received PCI as well.Compare the grade of NYHA ,the size of left ventricle and the ejection fraction r between two groups3 months late.Results There were no deference between groups in the grade of NYHA,the size of left ventricle and the ejection fraction at the beginning of treatment,but the grade of NYHA,the size of left ventricle and the ejection fraction in PCI group were better than contrast group 3 months later(P

ObjectiveTo find out if the immune system derived thyroid stimulating hormone(TSH) β splice variant(TSHβ-Ⅴ) would be regulated by circulating thyroid hormone levels to get a further understanding of the function and mechanism of this TSHβ-Ⅴ in thyroid homeostasis.MethodsA total of 20 weaning Balb/c mice (half male and half female) were selected and randomly divided into two groups according to their body mass and gender(n =10).Mice of control group were fed with common diet and deionized water.Mice of the low-iodine(LI) group were fed with low-iodine diet(containing iodine 20 - 40 μg/kg,iodine-intake about 0.25 μg/d) and deionized water.The experimental period was 3 months.At the end of the experiment,mice were executed and the blood was collected to observe the levels of TSH and thyroid hormone by chemiluminescence immunoassay (CIA) ; bone marrow (BM),peripheral blood(PBL),thyroid gland and pituitary were collected to assay the TSHβ-Ⅴ mRNA expression by real-time quantitative reverse transcriptase polymerase chain reaction (RT-PCR).ResultsThe serum free thyroxine(FT4) and total thyroxine(TT4) levels in LI group of mice[(0.47 ± 0.70)nmol/L,(2.41 ± 0.28)pmol/L] were significantly lower than that of the control group of mice [(55.2 ± 3.68) nmol/L, (32.72 ± 1.02) pmol/L,t =43.81,86.04 、all P ＜ 0.01 ] and the serum total triiodothyronine(TT3) and free triiodothyronine(FT3) reduction in LI group of mice[ (0.76 ± 0.08)nmol/L,(4.01 ± 0.40)pmol/L] were significantly lower than that of the control group of mice [ (1.10 ± 0.06)nmol/L,(5.40 ± 0.38)pmol/L,t =9.81,7.5 1,P ＜ 0.01 ].Iodine insufficiency strongly elevated the serum TSH in LI group of mice[ (35.67 ± 17.39)mU/L] than that in control group of mice[ (0.24 ± 0.10)mU/L,t =- 6.11,P ＜ 0.01 ].The mRNA levels of TSH β-Ⅴ in BM (9.62 ± 0.60) and in PBL( 9.25 ± 0.83 ) of LI group of mice were lower than those in control group of mice (7.69 ± 0.36,7.11 ± 0.41,t =6.77,5.64,P ＜ 0.01),while the mRNA level of TSH β-Ⅴ in pituitary of LI group of mice (1.99 ± 0.61) was increased compared with that in control group of mice (5.75 ± 0.98,t =- 8.02,P＜ 0.01).Compared with control group of mice(9.12 ± 0.62),the level of thyroid TSH β-Ⅴ mRNA in LI group of mice (9.32 ± 0.91 ) was not significantly changed (t =0.45,P ＞ 0.05).There was no detectable native TSHβ in BM,PBL and thyroid.The mRNA level of native TSHβ in pituitary in LI group of mice( - 7.17 ± 1.78) was dramatically elevated compared to that in control group of mice( - 1.43 ± 0.51,t =- 7.60,P ＜ 0.01 ).ConclusionsThe mRNA levels of TSHβ-Ⅴ are suppressed in BM and PBL in low iodinediet induced hypothyroidism mice,which suggest that immune system derived TSHβ-Ⅴ may be more important thannative TSHβ in immune-thyroid regulation.

Objective To investigate gene polymorphisms of drug targets and mutations associated with drug resistance in Pneumocystis jiroveci (P.jiroveci) isolates.Methods Among 148 samples isolated from human immunodeficiency virus (HIV)infected patients with pneumonia in Guangdong,mitochondrid larg subunit rRNA (mtLSUrRNA) gene was amplified from 51 samples.Dihydropteroate synthase (DHPS),dihydrofolate reductase (DHFR) and Cytochrome b (CYB) genes of P.jiroveci were detected by gene sequencing,and compared with the reference sequences in GenBank to evaluate gene polymorphisms.Results P.jirovecii DHPS,DHFR and CYB genes were all successfully amplified from 51 samples.For DHPS gene,48 (94.1％) were wild-type and 3 (5.9％) had gene mutation associated with drug resistance.For DHFR gene,30 were wild-type,and 21 had a synonymous mutation at position 312,and 1 nonsynonymous mutation at position 188.There were no mutations associated with drug resistance.For CYB gene,polymorphisms of were detected at 5 sites,4 of which were synonymous mutations,1 was non-synonymous mutation.No mutation associated with drug resistance was found.Based on the gene polymorphism of CYB6,the strains can be classified into 6 genotypes,and 2 were first detected,including 25 CYB1,13 CYB2,2 CYB5,4 CYB8,as well as newly detected 4 CYB10 and 3 CYB11 strains.Conclusions The mutations associated with drug resistance in P.jiroveci isolates in Guangdong remain uncommon.CYB gene shows gene polymorphisms and can be selected as one of targeted genes for multilocus sequence typing.

Objective To investigate the role of hemoglobin (Hb),red blood cell count (RBC) and hematocrit (HCT) during the first trimester in predicting the occurrence of gestational diabetes mellitus (GDM) in the third trimester in nonanemic women in Kunming.Methods A prospective study was carried out.Routine blood tests were performed in 1189 nondiabetic singleton pregnant Chinese women at their initial prenatal healthcare before 14 gestational weeks in the First Affiliated Hospital of Kunming Medical University from January 1,2008 to December 31,2009.They were divided into four groups by quartiles of Hb,RBC and HCT levels respectively.Differences of GDM incidence at the third trimester were compared among groups.Kollmogorov Smirnov test was used to detect normal distribution of Hb,RBC and HCT results.Differences between rates of groups were detected by Chi-square test.Independent risk factors of GDM were screened by Logistic regression analysis.Results The incidence of GDM at the third trimester increased as quartile categories of Hb,RBC and HCT ascended.The GDM incidence (13.1 ％,34/259) of ≥75th Hb group was higher than that of ＜ 25th group (5.9 ％,18/304,x2 =8.53,P＜ 0.0 1),25th ～ group (9.2 ％,30/326,x2=4.18,P＜0.05) and 50th～ group (9.3％,28/300,x2=3.96,P＜0.05).The GDM incidence (11.7 ％,34/290) of 50th～ RBC group (RBC≥4.44 × 1012/L) was higher than that of ＜25th group (6.3％,19/304,x2=5.30,P＜0.05),25th～ group (7.0％,21/298,x2 =3.93,P＜0.05); and the GDM incidence (12.1％,36/297) of ≥75th group was also higher than that of ＜25th group (6.3％,19/304,x2 =6.49,P＜0.05),25th～ group (7.0％,21/298,x2=3.85,P＜0.05).The GDM incidence of ＜25th HCT group (5.3％,16/303) was lower than that of 25th～ group (9.3％,29/311,x2=7.04,P＜0.01),50th～ group (10.8％,31/287,x2=3.93,P＜0.05) and ≥75th group (11.8％,34/288,x2=6.49,P＜0.05).Logistic regression analysis showed that Hb (OR=1.031,95％ CI:1.010-1.052),RBC (OR=2.286,95％CI:1.318-3.963) and HCT (OR=1.106,95％ CI:1.037-1.179) were independent risk factors of GDM (all P＜0.05).Conclusions High maternal Hb,RBC and HCT levels in the first trimester could be used in predicting GDM in pregnant women in Kunming area.

Corneal opacity is one of the most commonly used parameters for estimating postmortem interval (PMI). This paper proposes a new method to study the relationship between changes of corneal opacity and PMI by processing and analyzing cornea images. Corneal regions were extracted from images of rabbits' eyes and described by color-based and texture-based features, which could represent the changes of cornea at different PMI. A KNN classifier was used to reveal the association of image features and PMI. The result of the classification showed that the new method was reliable and effective.

Objective To appraise the analytical capability of flow cytometric bead array for lung cancer markers through the tests of limit of detection,relative standard deviation,specificity,methods comparation and linearity rang.Methods The limit of detection,relative standard deviation,specificity and linearity rang in detection of Carcinoembryonic antigen (CEA),cytokeratin 19 (Cyfra21-1) and neuron specific enolase (NSE) in serum were evaluated by flow cytometer.Western blotting method was ultilized to validate the specificity of antibody-antigen recognization.The interference of hemoglobin,three acyl glycerol and bilirubin on the detection of CEA,Cyfra21-1 and NSE was tested.Compared to electrochemiluminescence immunoassay,the relative error for flow cytometric bead array was assessed.Results Flow cytometric bead array demonstrated that the limit of detection was 1.71 pg/mL for CEA,3.97 pg/mL for cyfra21-1,and 2.27 pg/mL for NSE.The relative standard deviation for intra-assay and inter-assay were below 10％ and 15％,respectively.The pair of antibodies can defferentially recognize antigens.The measurement for CEACAM6,CK18,NSE appeared that there was no significant cross-talking reaction.Three acyl glycerol and bilirubin did not significantly interfere with the detection for serum samples.Hemoglobin of 500 ng/mL can significantly interfere with the detection of Cyfra21-1 (P ＜ 0.05) and NSE (P ＜ 0.05).The correlation coefficient between flow cytometric array and electrochemiluminescence immunoassay was 0.984 2 for serum CEA,0.962 2 for serum cyfra 21-1 and 0.982 0 for serum NSE.The linearity ranged from 355.76 pg/mL to 367.74 ng/mL for CEA,from 87.89 pg/mL to 107.8 ng/mL for cyfra21-1,and from 90.12 pg/mL to 86.07 ng/mL for NSE.Conclusion Flow cytometric array for lung cancer markers may be of use in clinical detection.

Objective To prepare,purify and validate specific monoclonal antibodies(McAb) against fragment in C-terminal telopeptides of collagen type Ⅱ(EKGPDP)for clinical diagnostics and related research of osteoarthritis(OA).Methods 8 BALB/c mice were immunized with EKGPDP-KLH antigen complex.McAb against EKGPDP fragements were prepared by hybridoma technique.Immunoglobulin classes and subclasses were determined using an Immuno-Type~(TM)mouse McAb isotyping kit.Ascites were producted and McAb were purified by saturation ammonium sulfate(SAS)precipitation and protein G chromatography.Specificity and immunoactivity of antibodies were detected by indirect enzyme-linked imrnunosorbant assay(ELISA).Cartilage specimens were analyzed by immunohistochemical method.Results The hybridoma cells were obtained.10 IgG and 3 IgM single colonies were picked out by limiting dilution and ELISA kit.The titers of McAb in ascites were from 2.8?10~4 to 5.1?10~5.The McAb showed the characteristics of no cross reactions with KLH,BSA,cell culture supernatant,type Ⅰ,Ⅲ collagens and whole type Ⅱ collagen.The method of SAS could get better recovery,immunoactivity of the McAb than protein G chromatograply(t=25.26;P

BACKGROUND:Previous studies have confirmed that ethanol can promote adipogenic differentiation of bone marrow mesenchymal stem cells (BMSCs), and up-regulate the expression of PPARγ and aP2 in the tumor necrosis factor α (TNF-α) signaling pathway. As a member of the ZIP protein family, Zrt/Irt-like protein 1 (ZIP1) is closely related to bone metabolism and osteogenic differentiation.OBJECTIVE:To study the effect of BMSCs transfected by ZIP1 on TNF-α signaling pathway in the process of adipogenic differentiation.METHODS:The BMSCs from rabbits were isolated and cultured under different concentrations of alcohol (0.03, 0.09,0.15, 0.21 mol/L), followed by transfection by ZIP1 siRNA and ZIP1 expression vector.RESULTS AND CONCLUSION:After culture in alcohol, the expression levels of aP2 and PPARγ proteins were both significantly increased (P < 0.05), and the level of triglyceride was increased in all alcohol groups except for 0.03 mol/L alcohol group (P < 0.05). After siRNA transfection, the expression levels of aP2 and PPARγ as well as the level of triglyceride were increased significantly in all the alcohol groups (P < 0.05); however, ZIP1 transfection decreased the expression levels of aP2 and PPARγ proteins (P < 0.05). To conclude, ZIP1 siRNA could promote the adipogenic differentiation of BMSCs through the activation of TNF-α signaling pathway.

CD4 Antigens ; blood ; CD4-CD8 Ratio ; CD8 Antigens ; blood ; Child, Preschool ; Female ; Humans ; Infant ; Male ; Seizures, Febrile ; immunology