Such parameters of TiO2 nanoparticle by Sol-gel method are analyzed through X-ray and SEM as the gelation time,crystal structure,particle size and morphology.

OBJECTIVE:To investigate the effects of long-term use of calcium channel blocker-Diltiazem(Dil)on the dosage of ciclosporin A and renal function of renal graft recipients.METHODS:Dil was administed in67renal graft recipi?ents,meanwhile who were orally taking CsA with another59renal graft recipients served as controls.The dosages of ci?closporin A of2group were adjusted to the level within therapeutic window,then the dosage of CsA and serum creatinine change of the2groups36mo after drug administration were observed.RESULTS:12mo,24mo and36mo after operation,the synchronized cyclosporin A dosages in Dil group were lower than the control group respectively by14353mg,9656mg and7817mg.No significant differences were found in serum creatinine levels between the2groups within the first12mo after operation.Thereafter,the creatinine levels in the control group has a faster increase and the creatinine level in Dil group was significantly lower than that of the control group18mo～36mo after operation(P

OBJECTIVETo isolate and identify the pathogen of Dengue fever from Shenzhen city in 2005 - 2006, and to analyze the molecular characteristics of the isolated Dengue virus strain as well as to explore its possible origin.

METHODSIgM and IgG of serum samples taken from 60 suspected Dengue fever patients were detected by ELISA and immunochromatography, and 9 specimens were positive. Nine samples from patients with early stage Dengue fever were used to isolate virus with C6/36 cell line and the positive cell cultures were identified by MGB fluorescent PCR. The type of isolated virus strain was determined by RT-semi-nested-PCR and fluorescent PCR. E gene of isolated virus strain was amplified by RT-PCR and sequenced. Homology and phylogenetic tree of E gene of Shenzhen Dengue virus with the strains isolated from other areas were constructed.

RESULTSOf nine antibody-positive serum samples, one strain of Dengue virus was successfully isolated. The isolated virus strain was confirmed as Dengue virus type 2 and designated as DEN2-SZ0521. The homology of nucleotide sequence and the deduced amino acid sequence of E gene of SZ0521 with standard type 2 Dengue virus NGC strain was 94.2% and 98.2%, but the homology with standard Dengue virus 1, 3, 4 in the same fragment were 59.1%, 57.2%, 58.5% and 68.1%, 66.7%, 63.2%, respectively. The phylogenetic tree indicated that SZ0521 had the greatest similarity with the Malay0412a/Tw strain and they lied in the same branch of the phylogenetic tree. The corresponding homology of nucleotide sequence and amino acid sequence was 99.8% and 100%, respectively. The isolated Dengue virus type 2 belonged to genotype IV with Indonesia-76, Somalia-84 and Sri Lanka-90.

CONCLUSIONDengue virus was isolated from Shenzhen for the first time, and it was classified as type 2. It was confirmed that the type 2 Dengue virus may come from the epidemic area in Malaysia.

Aedes ; virology ; Animals ; China ; Dengue ; virology ; Dengue Virus ; classification ; genetics ; isolation & purification ; Genes, Viral ; Humans ; Phylogeny ; Sequence Analysis, Protein ; Sequence Analysis, RNA

OBJECTIVETo clone tpn17 and tpn47 genes of Treponema pallidum and then construct their prokaryotic expression systems,to establish ELISAs based on rTpN17 and rTpN47 as antigens and to evaluate the sensitivity and specificity of the ELISAs for detection of serological diagnosis of syphilis.

METHODSThe whole length of tpn17 and tpn47 genes was amplified by PCR and then their prokaryotic expression systems were constructed. SDS-PAGE was used to measure the expression of the target recombinant proteins rTpN17 and rTpN47. Ni-NTA affinity chromatography was applied to extract rTpN17 and rTpN47, while Western blot was performed to determine the specific immunoreactivity of rTpN17 and rTpN47. By using rTpN17 and rTpN47 as the coated antigen, respectively, ELISAs (rTpN17-ELISA and rTpN47-ELISA) were established to detect serum samples from 200 healthy individuals, 25 RA patients, 17 SLE patients and 211 syphilis patients. The detection effects of the ELISAs were compared to those of TRUST and TPHA.

RESULTThe sequence similarity of the cloned tpn17 and tpn47 genes was 100 % compared with the corresponding sequences in GenBank. The expression outputs of rTpN17 and rTpN47 were approximately 37.2 % and 26.8 % of the total bacterial proteins, respectively. Both the extracted rTpN17 and rTpN47 could take place remarkable conjugation reactions to the sera with positive antibody against Treponema pallidum.The positive detection rate of TPHA (99.1%) was the highest (P<0.001). The positive detection rates of rTpN17-ELISA (85.3 %) and rTpN47-ELISA (84.3 %) were similar (P>0.05). The positive detection rates of TRUST (72.5 %) was lower than that of rTpN17-ELISA (P=0.001) but similar to that of rTpN47-ELISA (P=0.014). The detection results of all the serum samples from healthy individuals, RA patients and SLE patients were negative, whereas 7.1 % (3/42) of the samples from RA or SLE patients were positive.

CONCLUSIONrTpN17 and rTpN47 are still maintaining their original immunoreactivity. The ELISAs using rTpN17 or rTpN47 as the antigen are rapid, simple and convenient, higher sensitivity and specificity methods for serological screening and detection of syphilis.

Antibodies, Bacterial ; Antigens, Bacterial ; Blotting, Western ; Enzyme-Linked Immunosorbent Assay ; methods ; Female ; Humans ; Male ; Syphilis ; diagnosis ; Syphilis Serodiagnosis ; Treponema pallidum ; chemistry ; immunology ; isolation & purification

OBJECTIVE: To investigate the changes of ryanodine receptor 2 (RyR2) mRNA expression in rats suffering from acute myocardial ischemia. METHODS: SD rats were divided randomly into normal control group, myocardial ischemia group and sudden death group. The models of myocardial ischemia and sudden cardiac death were induced by intraperitoneal injection of hypophysine. The changes of RyR2 mRNA expression in cardiac sarcoplasmic reticulum (SR) of rats suffering from myocardial ischemia were detected by fluorescent RT-PCR technique. RESULTS: The levels of RyR2 mRNA in the myocardial ischemia group and sudden death group were significant lower than those in the control group (P<0.05). CONCLUSION Myocardial ischemia may induce down-regulation of cardiac SR RyR2 mRNA expression.
Animals ; Death, Sudden, Cardiac ; Down-Regulation ; Female ; Forensic Pathology ; Male ; Myocardial Ischemia/metabolism* ; RNA, Messenger/metabolism* ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction/methods* ; Ryanodine Receptor Calcium Release Channel/metabolism* ; Sarcoplasmic Reticulum/metabolism*

BACKGROUNDTo study oxidative stress in patients with chronic hepatitis B.

METHODSMalondialdehyde (MDA), total anti-oxidative ability and ascorbic acid were measured as markers of oxidative stress in 30 patients with chronic hepatitis B, besides HBV DNA and ALT.

RESULTSMDA was significantly higher in patients with hepatitis B than the controls (P less than 0.05). Ascorbic acid was significantly higher in patients with normal ALT than the controls (P less than 0.01). MDA was significantly higher in patients with increased ALT than the controls and in patients with normal ALT. MDA was significantly positively correlated with ALT (r=0.61), and ascorbic acid was significantly negatively correlated with ALT (r=-0.64) in patients with hepatitis B. No significant relationship was found between HBV DNA and other indices of oxidative stress. No significant difference in total anti-oxidative ability was found among all groups.

CONCLUSIONThere was a disturbance between oxidative stress and anti-oxidative ability in patients with chronic hepatitis B. In patients with increased ALT, oxidative stress became high. In patients with normal ALT, oxidative stress level was low. The indices of oxidative stress should be detected in patients with hepatitis B, in addition to HBV markers.

Adolescent ; Adult ; Alanine Transaminase ; blood ; Ascorbic Acid ; blood ; Child ; DNA, Viral ; blood ; Female ; Hepatitis B virus ; genetics ; Hepatitis B, Chronic ; physiopathology ; Humans ; Male ; Malondialdehyde ; blood ; Middle Aged ; Oxidative Stress ; physiology

OBJECTIVETo evaluate protein loss in critically ill patients with acute renal failure during continuous veno-venous hemofiltration (CVVH) and analysis the major factor impacting protein clearance.

METHODSA analysis was carried out in eighteen (twelve male and six female) sepsis or severe acute pancreatitis patients with acute renal failure from September 2008 to September 2009. The average age was 45 years (39 - 62 years). CVVH was conducted for 24 h in all patients. Effluent volume, blood speed, ultrafiltration rate and transmembrane pressure (TMP) were 4000 ml/h, (277 ± 89) ml/h, (179 ± 4) ml/min and (173 ± 48) mm Hg (1 mm Hg = 0.133 kPa) respectively. Blood samples were collected before and after filtration in order to detect protein concentration. Ultrafiltrate was obtained hourly to measure protein concentration and calculate protein loss during session.

RESULTSMean protein concentration was (231 ± 67) mg/L and protein loss was (22 ± 6) g/d in ultrafiltrate samples. The difference in serum protein level during hemofiltration was not significant [(56 ± 6) g/L vs. (55 ± 10) g/L, P > 0.05], while there was a weak, but statistically significant correlation between the ultrafiltrate protein concentration and the corresponding value for serum protein (r = 0.481, P < 0.05). However, there was a strong and statistically significant correlation between the ultrafiltrate protein concentration and the TMP (r = 0.564, P < 0.01). Stepwise multiple regression analysis showed that TMP and serum protein concentration played a pivotal role in ultrafiltrate protein loss.

CONCLUSIONSIn addition to renal replacement therapy, serum protein would be cleared through hemofilter during CVVH. TMP and serum protein concentration are the main factors that affect protein loss in ultrafiltrate. As a result, it is necessary to take account of the protein loss in ultrafiltrate when setting nutritional schedule.

Acute Kidney Injury ; therapy ; Adult ; Blood Proteins ; deficiency ; Critical Illness ; Female ; Hemofiltration ; adverse effects ; Humans ; Male ; Malnutrition ; etiology ; Middle Aged

The aim of this study was to examine the priming effect of sphingosine 1-phosphate (S1P) on fMLP-activated neutrophils, mainly to detect the neutrophil respiratory burst products, and to investigate the signaling pathway involved in S1P activity. Flow cytometry was used to evaluate the new isolated neutrophil; the superoxide anion output was detected indirectly by cytochrome C reduction in respiratory burst; the dihydro-rhodamine 123 was used to detect the intensity of respiratory burst; the signal transduction pathways of neutrophil respiratory burst were explored by Western blot. The results showed that after pretreated with S1P, the level of superoxide anion released by fMLP-activated neutrophils significantly increased; the Rhodamine 123 mean fluorescence intensity in S1P primed fMLP-activated neutrophils group was significantly higher than that in fMLP treatment group; PI3K and Akt proteins involved in the signal pathway of neutrophil respiratory burst. It is concluded that S1P is a new priming reagent, which primes respiratory burst of fMLP-activated neutrophils; this signal pathway may be that S1P interacts with its receptor, activates PI3K, then activates Akt-transmitting signals through NADPH oxidase, finally results in the respiratory burst.
Cells, Cultured ; Humans ; Lysophospholipids ; metabolism ; NADPH Oxidases ; metabolism ; Neutrophils ; metabolism ; physiology ; Proto-Oncogene Proteins c-akt ; metabolism ; Receptors, Lysosphingolipid ; metabolism ; Respiratory Burst ; Signal Transduction ; Sphingosine ; analogs & derivatives ; metabolism ; Superoxides ; metabolism

To explore the protective effect of Angelica sinensis polysaccharides(ASP) on subacute renal damages induced by D-galactose in mice and its mechanism. Male C57BL/6J mice were randomly divided into 3 groups, with 10 mice in each group. The D-galactose model group was subcutaneously injected with D-galactose (120 mg x kg(-1)), qd x 42; the ASP + D-galactose model group was intraperitoneally injected with ASP since the 8th day of the replication of the D-galactose model, qd x 35; and the normal control group was subcutaneously injected with saline at the same dose and time. On the 2nd day of after the injection, the peripheral blood was collected to measure the content of BUN, Crea, UA, Cys-C; paraffin sections were made to observe the renal histomorphology by HE staining; senescence-associated β-g-alactosidase (SA-β-Gal) stain was used to observe the relative optical density (ROD) in renal tissues; transmission electron microscopy was assayed to observe the renal ultrastructure; the renal tissue homogenate was prepared to measure the content of SOD, GSH-PX, MDA; the content of AGEs and 8-OH-dG were measured by ELISA. According to the result, compared with the D-galactose model group, the ASP + D-galactose model group showed obviously decreases in the content of BUN, Crea, UA, Cysc, AGES, 8-OH-dG, the number of hardening renal corpuscle, renal capsular space and renal tubular lumen, ROD of SA-β-Gal staining positive kidney cells, mesangial cells, basement membrane thickness, podocyte secondary processes fusion and MDA and increases in the number of normal renal corpuscle, ribosome and rough endoplasmic reticulum in podocytes, the activity of SOD and GSH-PX. In Conclusion, A. sinensis polysaccharides can antagonize kidney subacute damages induced by D-galactose in mice. Its protective mechanism may be correlated with the inhibition of the oxidative stress injury.
Angelica sinensis ; chemistry ; Animals ; Deoxyguanosine ; analogs & derivatives ; metabolism ; Drugs, Chinese Herbal ; administration & dosage ; Galactose ; adverse effects ; Humans ; Kidney ; anatomy & histology ; drug effects ; injuries ; Kidney Diseases ; chemically induced ; drug therapy ; metabolism ; prevention & control ; Male ; Mice ; Mice, Inbred C57BL ; Oxidative Stress ; drug effects ; Polysaccharides ; administration & dosage ; Protective Agents ; administration & dosage

To confirm the mechanism of exosomes as tumor vaccines inducing immunity response, dendritic cells (DCs) were induced from human peripheral blood mononuclear cells, while exosomes were isolated from DC loaded tumor antigen. The effect of exosomes on priming T cell proliferation was analysed under conditions with or without DCs, or DCs at different mature stages. The function of exosomes in immunity was detected through block test after blocking some molecules (CD11a, CD11b, CD11c, CD54, MFG-E8 and CD83). The effect of DCs on embedded exosomes was observed by confocal microscopy, the effect of blocking surface molecules on exosomes on DC-embedding exosomes was assayed by flow cytometry. The results indicated that both exosomes derived from imDC (imDex) and exosomes derived from mDC (mDex) could not prime T cells without DC or with imDC. The exosomes derived from mDC induced with different cytokines (LPS, TNF-alpha, CpG, CD40L) were no significant difference in concentrations but were different in effect. The immunity function of exosomes depended on CD11a, CD11b, CD11c, CD54, MFG-E8 and CD83 molecules, the effect of priming T cells is reduced when these molecules were blocked. Confocal microscopy and FACS assay showed that blocking CD11a and CD54 could inhibit exosome-targeted DC and DC-embedded exosomes. It is concluded that the exosomes target DCs through their surface molecules, therefore results in immune response of T cells.
Antigens, Neoplasm ; immunology ; Cells, Cultured ; Dendritic Cells ; cytology ; immunology ; secretion ; Exosomes ; immunology ; Humans ; K562 Cells ; Lymphocyte Activation ; drug effects ; T-Lymphocytes ; cytology ; immunology