Objective To know the expression of 47 kD protein of orientia tsutsugamushi(Ot) Karp strain and test its immunogenicity and antigenicity.Methods Forty-seven kD protein gene that code the whole mat peptide was amplified from the genomic DNA of Ot.Karp strain by PCR and constructed to a recombinant plasmid pGEX-47.The E. coli cells BL21 were transformed with pGEX-47,then the transformants were induced to express the recombinant 47 kD protein by IPTG. The antigenicity and immunogeni-(city) were identified by Dot Blot method and the experiment of immunizing mice respectively.Results 1. The PCR product of 47 kD protein gene of Ot.Karp strain was obtained and its sequence was the same as the published 47 kD protein gene.2. The E.coli expression plasmid pGEX-47 constructed was identified containing the 47 kD protein gene fragment by restrication analysis and PCR identification,and an expression band about 47 kD was detected by SDS-PAGE of the purified product from GST-fusson protein expressed by pGEX-47. 3.Dot Blot analysis and the experiment of immunizing mice testified its antigenicity and immunogenicity.Conclusions The 47 kD protein gene of Ot Karp strain is amplified that code the whole mat peptide, which can be expressed in prokaryotic cells and the expressed protein has antigenicity and immunogenicity.

Using colon adenocarcinoma cell line HT-29 as testing material, adhesion test was carried out for 24 strains of Bifidobacteria isolated from dejecta of longevous people aged one hundred years. The results showed that all strains could adhere to colon adenocarcinoma cells HT-29, among which the strains of TTF, Z2, TZ5 and (J-1) showed higher adhesive capability. Thus these four strains of Bifidobacteria were further tested for their endurance to stomach acid and bile acid. The production of vitamin B by these bacterial were also tested. All these tests indicated that the strain of TTF has high ability to produce Vitamin B_(1), B_(2), B_(6), B_(12), The survival rate after the treatment under pH 3.0 for 120 min was 93.11%, at the same time, the strain TTF can survive after being treated with 2% bile acid for 24h.

Castleman Disease ; complications ; metabolism ; pathology ; surgery ; Dendritic Cell Sarcoma, Follicular ; complications ; metabolism ; pathology ; surgery ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Receptors, Complement 3b ; metabolism ; Thoracic Diseases ; complications ; metabolism ; pathology ; surgery ; Thoracic Wall ; Vimentin ; metabolism

Nerve growth factor(NGF) was firstly discovered as a member of neurotrophin family,and the research and development of NGF has been lasting more than fifty years since its discovery.To this end,a two-step and high –yield chromatographic method which consists of cation ion-exchange chromatography and reversed-phase chromatography was reported to isolate recombinant human beta nerve growth factor(? –NGF) secreted by constructed Chinese hamster ovary cells(CHO/dhfr-) from the culture media.Through the process of purification,the purity of protein which was determined by SDS-PAGE and RP-HPLC has reached to 95%,and the recovery of ? –NGF routed by RP-HPLC could be 70%.Furthermore,the biological activity of final purified protein evaluated by PC12 cells and dorsal root ganglia(DRG) exhibited the same performance as the standard protein of ? –NGF bought from Sigma,which indicated that there is no loss of biological activity through the isolation process.The conclusion suggested that an economical isolation method of recombinant human ? –NGF could be practiced on the industrial process of purification.

This study is to investigate the effect of artesunate on transforming growth factor-beta1 (TGF-beta1) induced epithelial-mesenchymal transition (EMT) and its possible mechanism. After the in vitro cultured RLE-6TN cells were treated with TGF-beta1 then artesunate intervened on it, after 24 h, expression of the markers of mesenchymal cell was assayed using Western blotting and real-time PCR analysis. Western blotting was also used to detect the effect of TGF-beta1 on the Smad3 and Smad7 expressions of RLE-6TN cells. Morphological alterations were examined by phase-contrast microscope, and ultrastructure changes by electron microscope. Incubation of RLE-6TN cells with TGF-beta1 resulted in the up-regulation of the expression of the mesenchymal cell markers, after artesunate intervened on it, resulted in the down-regulation of the expression. Meanwhile, incubation with artesunate intervened on RLE-6TN cells could lead to the apparent down-regulation of the expression of Smad3 and up-regulation of Samd7 and the transition of RLE-6TN cells to mesenchymal-like by TGF-beta1 induction, after artesunate intervened on it, RLE-6TN cells to epithelial-like. TGF-beta1 induced epithelial-mesenchymal transition process; artesunate can inhibit TGF-beta1-induced epithelial-mesenchymal transition process, the possible mechanism is up-regulation of the expression of Smad7 and down-regulation of the expression of Smad3, meanwhile inhibits phosphorylation of Smad3.
Actins ; genetics ; metabolism ; Animals ; Artemisia ; chemistry ; Artemisinins ; isolation & purification ; pharmacology ; Cell Line ; Cell Proliferation ; drug effects ; Epithelial Cells ; cytology ; metabolism ; Epithelial-Mesenchymal Transition ; drug effects ; Idiopathic Pulmonary Fibrosis ; pathology ; Plants, Medicinal ; chemistry ; Pulmonary Alveoli ; cytology ; RNA, Messenger ; metabolism ; Rats ; Smad3 Protein ; genetics ; metabolism ; Smad7 Protein ; genetics ; metabolism ; Transforming Growth Factor beta1 ; pharmacology ; Vimentin ; genetics ; metabolism

OBJECTIVE To investigate the protective effect of curcumin on Aβ1-42 damaged cells. METHODS SH-SY5Y cells were cultured with Aβ1-4210μmol · L-1 in the absence or presence of curcumin 1, 5 or 10 μmol · L-1. Cell viability was assayed by MTT. Cell membrane damage was detected by the concentration of lactate dehydrogenase (LDH) in culture medium. Cell apoptosis was measured by flow cytometry with Annexin Ⅴ-FITC/PI staining. Mitochondrial membrane potential was characterized by fluorescence of JC-1 dye. Enzymatic activity of caspases-9 and-3 was measured by colorimetric assay. Protein expression of caspase-3 was detected by Western blotting. RESULTS Compared with vehicle control, the cell viability, concentration of LDH and both early and late apoptosis in Aβ1-4210 μmol · L-1 damaged group were decreased(P<0.01). However, the cell viability, release of LDH and both early and late apoptosis in curcumin group were promoted compared with that in Aβ1-4210μmol·L-1 damaged group. Curcumin inhibited Aβ1-42-induced depolarization of mitochondrial membrane potential(P<0.01), and attenuated Aβ1-42-induced activation of both caspases9 and caspases3 in a concentration-dependent manner, respectively(r=0.990, P<0.01; r=0.996, P<0.01). There were no significant differences in the above detected indexes between curcumin 10 μmol · L-1 group and vehicle control group. CONCLUSION Curcumin inhibits Aβ1-42-induced cell damage and apoptosis by promoting mitochondrial membrane potential and depressing the activation of caspases.

Objective To compare the heart function and observe the clinical prognosis of dilated cardiomyopathy(DCM) in children treated with combination of prednisone and azathioprine and high dose intravenous immunoglobulin(IVIG).Methods From Jan.1998 to May 2004,20 children who met the diagnostic criteria of DCM were divided into 2 therapeutic groups(groupⅠ,groupⅡ) according to the time of hospitalization.On the basis of traditional congestive heart failure(CHF) therapy,the children in groupⅠwere treated with immunosuppressive therapy(prednisone plus azathioprine) and the children in groupⅡ were treated with high dose IVIG.The course of treatment was 6 months for all children.Heart functions in all children were evaluated and echocardiographic parameters including left auricular diameter(LA),left ventricular end diastolic diameter(LV),left ventricular ejection fraction(LVEF) and cardiac index(CI) were measured before and after therapy,respectively.All children were followed up for 1 year to observe survival rate(%).The findings were analyzed by t test and ?~2 test.Results The percentage of heart function increased from Ⅲ-Ⅳ toⅠ-Ⅱin groupⅠ,which was lower than that in groupⅡ(P

Objective To evaluate the accuracy of the fetal weight estimation system in terms of estimating the fetal weight.Methods The fetal weight estimation system developed by the Department of Electronic Engineering,Fudan University was applied to estimate the fetal weight in many hospitals,and the differences between the estimated results of the system and the actual weight of newborn fetuses,as well as the accuracy of the estimation system and the traditional estimation methods were compared.Results A total number of 610 cases were employed,with each case including the ultrasound images of fetus before birth,the corresponding measurement data and the newborn weight.The correlation coefficients between the weight of the newborn babies and the fetal parameters,which were measured by the estimation system in terms of the biparietal diameter,the head circumference,the abdomen circumference and the femur length,were higher than those obtained by manual annotations,with improvements of 0.08,0.17,0.09 and 0.05,respectively.The absolute error of the estimated weight of the system with respect to the actual weight was 44 g smaller than those estimated by traditional method,and the variance of the absolute error was 53 g smaller;while the corresponding absolute percentage error was 1.8％ lower,and the corresponding variance percentage was 1.5％ lower.Conclusions The fetal weight estimation system improves the accuracy of the prenatal ultrasound assessment and provides reliable information to clinical diagnosis.

Objective:To study the relationship between CYP4F2 gene polymorphism and coronary heart disease (CHD) in Mongolia patients ,and investigate clinical characteristics of these patients . Methods:All subjects received questionnaire . Gene amplification and genotyping were performed in 234 Mongolian CHD patients (CHD group) and 221 non-CHD pa‐tients (normal control group) using high temperature ligase detection reaction technique .The relationship between Mongo‐lian CHD and CYP4F2 gene polymorphisms of two single nucleotide polymorphism (SNP) sites (rs1558139 ,rs2108622) was analyzed .Results:Compared with normal control group ,there were significant rise in percentages of male (41.18% vs . 67.95% ) ,smoking history (32.13% vs .41.88% ) ,body mass index [BMI ,(21.66 ± 4.53 ) kg/m2 vs .(25.34 ± 5.37 ) kg/m2 ] and triglyceride level [ (1.66 ± 0.90) mmol/L vs .(1.92 ± 1.38) mmol/L] ,and significant reduction in level of high density lipoprotein cholesterol [ (1.18 ± 0.28) mmol/L vs .(1.07 ± 0.29) mmol/L] in CHD group , P<0.05 or <0.01.There were no significant difference in genotype and allele frequencies of rs 1558139 and rs2108622 between two groups . Conclusion:Clinical characteristics of Mongolian CHD patients include high male percentage ,smoking history ,high body mass index and high triglyceride level .CYP4F2 gene polymorphisms of rs1558139 and rs2108622 are not related to coronary heart disease in Mongolian patients .

Objective To explore the association of NH2-terminal pro-B-type natriuretic peptide ( NT-proBNP) with the risk of type 2 diabetes.Methods One hundred and twenty-six impaired glucose regulation( IGR) participants from Diabetic Identification Center of Tianjin Metabolic Diseases Hospital were included.NT-proBNP was measured in plasma samples collected from participants at baseline condition.Results At baseline, NT-proBNP was inversely associated with body mass index, waist circumference, fasting glucose, insulin and low-density lipoprotein-cholesterol( LDL-C) levels.During a follow-up of 2 years, 51 participants reported a new diagnosis of diabetes from OGTT.Baseline quartiles of NT-proBNP were inversely associated with diabetes risk, even after multivariable adjustment.Theadjustedrelativerisksfordiabeteswere1.0(reference),0.83(95%CI0.74-0.96),0.78(95%CI 0.68-0.90), 0.74 (95%CI 0.64-0.87) for the 1st, 2nd, 3rd, and 4th quartiles of baseline NT-proBNP, respectively ( P<0.01 ) .Conclus ion In IGRpopulation , lowlevels of NT-proBNP were associated with a significantly increased risk of type 2 diabetes.