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

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*

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

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

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

OBJECTIVETo investigate the relationship between alcohol consumption and risk factors for cardiovascular disease.

METHODSTwo hundreds and twenty six subjects were enrolled in the study and grouped to non-drinkers, mild drinkers, moderate drinkers and heavy drinkers. Serum GGT, hs-CRP, %CDT, HCY, lipoprotein were measured in all groups.

RESULTThere were significantly higher GGT levels with heavy drinkers than those with other groups (P <0.05), and GGT levels were increased with increasing alcohol intake; and there were significantly higher %CDT levels with heavy drinkers compared with those with no-drinkers; there was significant higher hs-CRP levels with heavy drinkers compared with those with mild and moderate drinkers (P<0.05); but in moderate drinkers there was significantly lower hs-CRP levels than non drinkers (P<0.05). Compared with non-drinkers, there were significantly lower LDL-C and TG levels with mild and moderate drinkers. There were no significant differences in CHOL, HDL-C, HCY, WBC, MCV levels among all groups. Heavy drinkers had higher smoking rate and higher prevalence of hypertension (P <0.05).

CONCLUSIONHeavy alcohol consumption results in increasing GGT,%CDT and hs-CRP and may increase cardiovascular disease risk along with other risk factors.Mild to moderate alcohol consumption is associated with lower hs-CRP concentration,which may protect the cardiovascular system through anti-inflammatory mechanism.

Adolescent ; Adult ; Aged ; Alcohol Drinking ; adverse effects ; blood ; Alcoholism ; blood ; C-Reactive Protein ; metabolism ; Cardiovascular Diseases ; epidemiology ; China ; epidemiology ; Humans ; Male ; Middle Aged ; Risk Factors ; Surveys and Questionnaires ; Transferrin ; analogs & derivatives ; metabolism ; Young Adult ; gamma-Glutamyltransferase ; blood

OBJECTIVETo study the effects of sodium hydrosulfide (NaHS), hydrogen sulfide (H2S) donor, on LPS-induced polymorphonuclear neutrophil (PMN) accumulation and its mechanism.

METHODSThe animal model of acute lung injury (ALI) caused by intravenous injection of lipopolysaccharides (LPS). Adult male Spraguce-Dawley (SD) rats were randomly divided into four groups (n = 8 - 12 per group): Control group (0.5 ml/kg normal saline i.v.), LPS-treated group (1 mg/kg, i.v.), LPS plus NaHS (1 mg/kg i.v. and 28 micromol/kg i.p., respectively) and NaHS group (28 micromol/kg i.p.). Animals were sacrificed at 6 h after agent administration. Morphological changes of lung tissues were observed and polymorphonuclear neutrophil (PMN) number in alveolar septum was tested. The apoptosis of PMN in the bronchoalveolar lavage fluid (BALF) was examined with in situ TdT-mediated dUTP end labeling (TUNEL). Intercellular adhesion factor-1 (ICAM-1) and nuclear factor-kappaB (NF-kappaB) expressions in the lung tissue were analyzed by Western Blot.

RESULTSThe results showed that bleeding, edema, PMN accumulation and other pathological signs in the lung tissue emerged after LPS injection. Compared to control rats, the LPS-treated rats had increased PMN number, decreased PMN apoptotic percentages, and increased expressions of ICAM-1 and NF-kappaB. Administration of NaHS into LPS-treated rats reduced the PMN number and expressions of ICAM-1 and NF-kappaB but increased PMN apoptotic percentages. In addition, NaHS alleviated the degree of ALI. There were no significant differences of the above indicators between NaHS-treated rats and control rats.

CONCLUSIONNaHS can reduce the PMN accumulation in the lung, and its mechanism is related to down-regulation expression of ICAM-1 and promotion of PMN apoptosis induced by inhibition of NF-kappaB pathway.

Acute Lung Injury ; chemically induced ; metabolism ; pathology ; Animals ; Apoptosis ; Hydrogen Sulfide ; pharmacology ; Intercellular Adhesion Molecule-1 ; metabolism ; Lipopolysaccharides ; adverse effects ; Lung ; drug effects ; metabolism ; pathology ; Male ; NF-kappa B ; metabolism ; Neutrophils ; cytology ; Rats ; Rats, Sprague-Dawley

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