There are two populations of bacteria to affect the formation of urinary stones in humanity. The first one can promote the formation of urinary stone by increasing urinary pH, decreasing concentration of urinary inhibitors , and damaging the protective urothelial glycosaminoglycan layer. The second inhibit the formation of urinary stones. These bacteria ( mainly the intestinal oxalate degrading bacteria such as Oxalobacter formigenes, lactic acid bacteria, Enterococcus faecalis etc) can decrease urinary oxalate concentration by regulating exogenous oxalate. The problems faced and the developing direction were also indicated.

Objective To determine the optimal procedure for preparation of the SPIO nanoparticles modified by dextran polylysine,and to evaluate the quality of this product.Methods The optimal preparation procedure of four factors (solution pH value,dextran concentration,reaction temperature and stirring rate) affecting SPIO hydrodynamic size were obtained through orthogonal experiments (3 levels).SPIO nanoparticles were synthesized under an optimum procedure.The shape and hydrodynamic size were detected by transmission electron microscope (TEM) and Malvern Zetasizer respectively.The content of Fe was detected by atomic absorption spectrophotometer,while chemical structures of SPIO nanoparticles were characterized and analyzed by infrared spectroscopy (IR) method.X-ray powder diffraction method was used to identify the ingredients,and the magnetic parameters were measured by vibrating sample magnetometer.Furthermore,experiments with ovarian cancer cells were performed to primarily validate the magnetic property of SPIO nanoparticles.Results The results of the orthogonal experiments showed that the optimum preparation procedure was as follows:dextran concentration of 10 mg/ml,pH 10,reaction temperature of 80 ℃ and stirring rate of 600 r/min.The TEM results showed the SPIO nanoparticles were in spherical shape,homogeneously distributed and uniform in size,and the mean diameter was 7.0 nm.The content of Fe was (12.36±0.08) g/L.The IR results clearly showed that the Fe3O4 was coated by dextran successfully.The Xray powder diffraction method showed that the sample contained Fe3O4 and the magnetism parameters indicated that the sample had superparamagnetism.The experiments with ovarian cancer cells demonstrated that SPIO nanoparticles could enter into the cells and then the cells had certain magnetic properties.Conclusions The SPIO nanoparticles synthesized under the optimal procedure are stable,small in size,with good dispersion and are feasible to enter into cells for rendering certain magnetic properties.This study has provided a good foundation and potential for further research.

AIM: To investigate the effects of sodium ferulate on cholesterol and triglyceride metabolism in atherosclerosis with hyperlipidemia.METHODS: The rabbit model of atherosclerosis was produced by feeding high lipid forages.RAW264.7 foam cell and HepG2 injured cell models were established by incubation with oxidized low density lipoprotein(ox-LDL).The atherosclerotic plaque area was measured,and serum lipids were detected.The cellular lipid accumulation was examined by oil red O staining.The cellular contents of total cholesterol and cholesterol ester were quantified by high performance liquid chromatography.The hepatic lipase(HL) mRNA expression was determined by RT-PCR.RESULTS:(1) Compared with hyperlipid group,the aorta atherosclerosis plaque area and the serum triglyceride level were significantly decreased in sodium ferulate-treated rabbits,but the serum cholesterol level showed little change.(2) Compared with ox-LDL group,the HL mRNA expression in HepG2 cells was enhanced significantly in sodium ferulate-treated group,but the cellular contents of total cholesterol and cholesterol ester in RAW264.7 foam cells showed little change.CONCLUSION: Sodium ferulate inhibits the formation of atherosclerotic plaque in high-cholesterol-fed rabbits aorta.This antiatherosclerotic function may reduce serum triglyceride level through enhancing the expression of HL mRNA without influencing serum cholesterol level and foam cell formation.

Objective To assess the application value of high time-resolved MR angiography (TR-MRA) in postoperative follow-up study of children with congenital heart diseases. Methods Seventy-three patients (median age 6 years, range 1-20 years) with congenital heart diseases who underwent TR-MRA scan after operation were retrospectively analyzed. Twenty-nine cases also were performed conventional contrast-enhanced MRA and forty-four cases were performed phase-contrast MRA. A 3D T1-weighted fast gradient-echo sequence was used for time-resolved three-dimensional MRA (10-20 dynamic data sets, less than three seconds per dynamic data set). The flow dynamics and morphology of pulmonary circulations, lung perfusion and collaterals flow direction were noted. All imaging quality was evaluated by using 5 scales. Left and right pulmonary artery flow volumes were measured and left and right pulmonary artery ratio was noted. SPSS22.0 was used in statistic analysis. The statistical analysis of comparing imaging quality was performed by using paired t-test. The intermodality agreement between TR-MRA and phase contrast in assessing left and right pulmonary perfusion was tested by Kappa coefficient. Results In 73 cases, imaging scores were over 3 and imaging quality was good enough for diagnosis. In 29 cases, there was no statistic difference between TR-MRA and conventional CE-MRA in demonstrating great vessels (P>0.05) except that CE-MRA scores(3.77 ± 0.39)was higher than TR-MRA scores(3.44 ± 0.55)of
inferior vena cava (IVC). There was statistic difference(t=3.68,P=0.01)between two sequences. TR-MRA could qualitatively demonstrate the pulmonary perfusion comparing to the results of PC. In PC sequence, there were 8 cases with symmetric and 36 cases with asymmetric left and right pulmonary perfusion. In TR-MRA sequence, there were 6 cases with symmetric and 38 cases with asymmetric left and right pulmonary perfusion. There was an excellent agreement between PC and TR-MRA (Kappa=0.83,P=0.01). Conclusions TR-MRA not only supplies with high spatial resolution imaging which demonstrates postoperative great arteries anatomy and also with high temporal resolution imaging which can demonstrate the preferential or balanced pulmonary blood flow and collaterals flow direction. TR-MRA is a very important sequence in follow-up study of congenital heart disease.

OBJECTIVETo prepare the superparamagnetic iron oxide (SPIO)-labeled antisense oligodeoxynucleotide (ASODN) probe and evaluate the application of this probe in cellular magnetic resonance imaging (MRI).

METHODSWe prepared the SPIO-labeled ASODN probe using chemical cross linking method to conjugate SPIO to ASODN, detected its configuration by atomic force microscopy, determined the conjugating rate and biology activation by high performance liquid chromatography, and detected the stability by polyacrylamide gel electrophoresis. After that, we transfected the SK-Br3 oncocytes which had over-expression of the c-erbB2 oncogene by this probes, observed the intracellular iron distribution by optical microscope, measured iron content by atomic absorption spectroscopy, and observed the signal change by MRI.

RESULTSAtomic force microscope showed that the SPIO-labeled ASODN probe was mostly spherical and well-distributed, with a diameter of 25-40 nm and a conjugating rate of 100%. This probe had inhered biological activity and stability. In addition, light microscopy revealed an intracellular uptake of iron oxides in the transfected SK-Br3 oncocyte, and the iron content of the group of transfected SK-Br3 oncocytes was significantly higher than those of other contrast groups (all P < 0.01). MRI showed that transfected SK-Br3 oncocyte had the lowest signal among all other cells (all P < 0.05).

CONCLUSIONSWe prepared the SPIO-labeled ASODN probe successfully. It can effectively transfect SK-Br3 oncocyte and enter SK-Br3 oncocyte, and thus reduce the signal intension in MRI.

Cell Line, Tumor ; DNA, Antisense ; chemistry ; genetics ; Ferric Compounds ; chemistry ; Humans ; Magnetic Resonance Imaging ; Magnetics ; Molecular Probe Techniques ; Oligodeoxyribonucleotides ; chemistry ; genetics ; Oxyphil Cells ; chemistry ; Receptor, ErbB-2 ; analysis ; genetics

To study the effect of micro (mi)RNA on cellular proliferation induced by hepatitis B x protein, HBx, in human liver cells and to investigate the underlying molecular mechanism of this cancer-related effect. The human L02 hepatocyte cell line was stably transfected with HBx (L02/HBx) or an HBx mutant (L02/HBx-d382) that induces higher levels of cellular proliferation. The differential miRNA expression profiles were determined by microarray analysis and confirmed by real-time PCR. Two miRNAs, miR-338-3p and miR-551b, that were found to be significantly down-regulated in the L02/HBx-d382 cells were selected for further study and transfected individually into cells using the lipofectamine procedure. The cell survival rate was analyzed by MTT assay, and cell cycles were assessed by flow cytometry. Expressions of cyclinD1, cyclinG1, and E2F1 were assessed by real-time PCR and Western blotting. Compared with the microarray miRNA profile of L02/pcDNA3.0 cells, six miRNAs were up-regulated and five miRNAs were down-regulated in the L02/HBx-d382 cells, while four miRNAs were up-regulated and 12 were down-regulated in the L02/HBx cells. The microarray results were consistent with real-time PCR results. Transfection of miR-338-3p and miR-551b significantly inhibited the cell survival rates (P less than 0.001) and induced G0/G1 phase cycle arrest. According to MTT results: for L02/HBx-d382 cells, compared with lipofectamine or non-transfected (NC) controls, the t value of miR-338-3p was 10.402, 9.133 and the t value of miR-551b was 8.763, 7.403; for L02/HBx cells, compared with lipofectamine or NC controls, the t value of miR-338-3p was 9.105, 8.074 and the t value of miR-551b was 7.673, 7.52. According to flow cytometry results: for L02/HBx-d382 cells, compared with lipofectamine or NC controls, the t value of miR-338-3p was 12.173, 11.107 and the t value of miR-551b was 15.364, 13.377; for L02/HBx cells, compared with lipofectamine or NC controls, the t value of miR-338-3p was 15.416, 13.378, and the t value of miR-551b was 13.276, 13.109. The protein levels of cyclinD1, cyclinG1, and E2F1 were significantly reduced by both miR-338-3p and miR-551b ( P less than 0.001). For L02/HBx-d382 cells, compared with lipofectamine or NC controls: E2F1 had t = 11.132, 10.031 and 12.017, 10.973, respectively; cyclinD1 had t = 15.654, 15.013 and 15.447, 14.733, respectively; cyclinG1 had t = 8.017, 7.661 and 7.402, 7.417, respectively. For L02/HBx cells, compared with lipofectamine or NC controls: E2F1 had t = 14.244, 13.331 and 15.022, 14.468, respectively; cyclinD1 had t = 8.695, 8.137 and 7.877, 7.503, respectively; cyclinG1 had t = 7.73, 7.471 and 7.596, 7.41, respectively. In contrast, the mRNA levels for E2F1, cyclinD1, and cylcinG1 showed no significant differences between the miRNA transfected cells and controls. Wild-type HBx and the high proliferation-inducing mutant HBx can influence the miRNA expression profile of L02 cells. HBx down-regulates miR-338-3p and miR-551b in L02 cells, and the high proliferation-inducing mutant has a more robust effect. The mechanism of miR-338-3p- or miR-551b-mediated cell growth inhibition appears to be related to the direct modulation of cyclinD1, cyclinG1, and E2F1.
Blotting, Western ; Carcinoma, Hepatocellular ; genetics ; metabolism ; pathology ; Cell Cycle ; Cell Line ; Cell Proliferation ; Cyclins ; genetics ; metabolism ; Gene Expression Regulation, Neoplastic ; Genes, Viral ; Hepatitis B virus ; genetics ; metabolism ; Hepatocytes ; metabolism ; pathology ; Humans ; Liver Neoplasms ; genetics ; metabolism ; pathology ; MicroRNAs ; genetics ; metabolism ; Mutation ; Oligonucleotide Array Sequence Analysis ; RNA, Messenger ; genetics ; Real-Time Polymerase Chain Reaction ; Trans-Activators ; genetics ; metabolism ; Transfection

Animals ; Disease Models, Animal ; Liver ; parasitology ; Mice ; Mice, Inbred BALB C ; Schistosomiasis japonica ; immunology ; Thiourea ; analogs & derivatives ; pharmacology

OBJECTIVE: To evaluate the clinical significance of serum hepatitis C virus (HCV) core antigen detected by enzyme linked immunosorbent assay (ELISA). METHODS: The serum HCV core antigen, which was taken from 149 patients with chronic hepatitis C, 20 patients of chronic hepatitis B and 20 health volunteers, was detected by ELISA. Meanwhile, the serum HCV RNA was detected by RT-PCR, and anti-HCV was detected by ELISA. RESULTS: The qualitative HCV core antigen in the serum, which was take from 20 patients of chronic hepatitis B and 20 health volunteers, was negative.The positive percentage of HCV core antigen was 49.66% in the 149 sera of patients with chronic hepatitis C. The coincidence of detective results of HCV RNA and HCV core antigen was 54.36%, without significant difference (P>0.05). The positive percentage of HCV RNA and HCV core antigen in the 149 anti-HCV antibody positive sera samples were 55.03% (82/149) and 49.66% (74/149), respectively, and there was no significant difference (P>0.05). CONCLUSION The qualitative HCV core antigen detected by ELISA has a high specificity. The positive percentage of HCV core antigen in the serum of patients with chronic hepatitis C is 49.66%. HCV core antigen is related to HCV RNA. HCV core antigen may be a useful serum marker which could show HCV viraemia like HCV RNA.
Hepatitis C Antigens ; blood ; Hepatitis C, Chronic ; blood ; Humans ; RNA, Viral ; blood ; Viral Core Proteins ; blood

OBJECTIVE: To study the association between S100A8 expression and prognosis in children with acute lymphoblastic leukemia (ALL). METHODS: The clinical data of 377 children with ALL who were treated with the CCLG-2008-ALL regimen were retrospectively reviewed. ELISA and PCR were used to measure serum protein levels and mRNA expression of S100A8. The Kaplan-Meier method was used for survival analysis and a Cox regression analysis was also performed. RESULTS: The children were followed up for 56 months, and the overall survival rate of the 377 children was 89.1%. The prednisone good response group had significantly lower S100A8 protein and mRNA levels than the prednisone poor response group (P<0.01). In the children with standard or median risk, both S100A8 protein and mRNA levels were associated with event-free survival rate (P<0.05). There were significant differences in S100A8 protein and mRNA levels between the children with different risk stratifications (P<0.01). The children who experienced events had significantly higher S100A8 protein and mRNA levels than those who did not (P<0.01). The Kaplan-Meier survival analysis and the Cox regression model suggested that S100A8 overexpression was an independent risk factor for the prognosis of children with ALL. CONCLUSIONS High S100A8 expression may be associated with the poor prognosis of children with ALL and is promising as a new marker for individualized precise treatment of children with ALL.
Calgranulin A ; metabolism ; Child ; Disease-Free Survival ; Humans ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; Prognosis ; Retrospective Studies

OBJECTIVETo elucidate the relationship between azoospermia factor(AZF) microdeletion of Y chromosome and azoospermia, the exact breakpoint of a severe Y-chromosome deletion was determined according to the physical map of AZF.

METHODSMultiplex polymerase chain reaction was used to amplify fifteen sequence tagged sites (STS), namely sY82, sY84, sY86 in AZFa, sY124, sY127, sY128, sY133, sY134, sY143 in AZFb, sY239, sY242 sY254, sY255 in AZFc, and sY145, sY152 in AZFd; sex-determining region Y(SRY) was taken as an internal control. And then sY82,sY86,sY85,sY84 were further analyzed using the sample of the patient who had Y-chromosome deletion by G band analysis to map the breakpoint at molecular level.

RESULTSAll 15 STS and sY85 were amplified in positive control while only sY82, sY86 were amplified in the clinical sample, thus the breakpoint was found to be between sY86 and sY85.

CONCLUSIONThis study on the patient provided the direct biomolecular evidence of the exact breakpoint of the severe Y-chromosome deletion and established the deletion map of acrocentric chromosome. It also proved that the patient's azoospermia was due to the deletion of AZF.

Azoospermia ; diagnosis ; genetics ; Chromosome Breakage ; Chromosome Deletion ; Chromosomes, Human, Y ; genetics ; Humans ; Male ; Polymerase Chain Reaction ; methods ; Reproducibility of Results ; Sensitivity and Specificity