Objective To further investigate the molecular mechanism of microRNA(miRNA)-24 on endothelial nitric oxide synthase(eNOS)gene expression, activity and the impact on its metabolites. Methods The plasmid of miRNA-24 was constructed and transfected to human umhilical vein endothelial cells(HUVECs). The proliferation of cells was detected by MTT test. Migration ability was measured by scratch wound model. The mRNA transcription and protein expressions of eNOS were determined by RT-PCR and Western blot respectively. The eNOS activity was detected by ELISA. The metabolite NO production in cell culture supernatant was detected by nitrate reduction. Results Compared with control group, the proliferation of endothelial cells in the miRNA-24 group was decreased by 54.32%;the migration of HUVECs was reduced by 48.62%;the expressions of eNOS mRNA and protein were de? creased by 43.92% and 42.71%, respectively. miRNA-24 suppressed eNOS activity by 73.20%. Meanwhile, the NO production was decreased by 55.29%. Conclusion miRNA-24 inhibits the synthesis and release of metabolite NO, which may be a molecular target of prevention and treatment in cardiovascular diseases.

Objective To evaluate the roles of magnetic resonance imaging and proton magnetic resonance spectroscopy(1H-MRS) in the diagnosis of meningiomas. Methods 98 patients with meningiomas underwent conventional pre-contrast MR and contrast MR. Among them, 28 cases had two dimensional single voxel or multi voxel 1 H-MRS simultaneously both in the lesion's region and the contralateral side. Results On precontrast MR images of 98 cases, T1 WI showed 58.1% (61/105) isointensities, 31.4% (33/105) faintly low intensities and 10. 5% (11/105) mixed intensities; T2WI showed 40. 0% (42/105) isointensities, 41.0%(43/105) hyperintensities, 10.5% (11/105) faintly low intensities and 8.5% (9/105) mixed intensities. After administration of Gd-DTPA, the solid part of the tumors exhibited various enhancement in all the 98 cases.28 cases of MRS exhibited specific different spectral peaks, including increased of choline-containing compounds(Cho), absent or decreased of acetylaspartate(NAA), and the unchanged of creatine(Cr). The value of NAA, Cr, Cho, NAA/Cr, Cho/Cr, NAA/Cho in the tumor center of meningioma were 0. 09 ± 0.06,0.31 ± 0. 22, 0.46 ± 0. 16, 0.33 ± 0. 42, 1.50 ± 0. 68, 0. 15 ± 0.08, compared with the contralateral normal region, Cr has no significant difference (P ＞ 0. 05), NAA, Cho, NAA/Cr, Cho/Cr, NAA/Cho had significantly differences(P ＜ 0.05). Conclusion Conventional pre-contrast MR and contrast MR is the most important dignostic means for meningiomas, 1H-MRS combined with MRI can improve the diagnostic accuracy of meningiomas.

OBJECTIVETo investigate the nephrotoxic effects of methyl cantharidimide tablets on urinary protein and enzymes in Beagle dogs.

METHODBeagle dogs were randomly divided into negative control group(blank tablet), methyl cantharidimide tablets group (6.11,12.21, 24.42 mg x kg(-1)), continuously 30 days of oral adminiStration, once a day. The drug and control group were collected and determined fresh urine in 1, 2, 3 and 4 weeks of the administration; Serum urea nitrogen (BUN), creatinine (Crea), total protein (TP) and albumin (ALB) as well as sodium, potassium, chloride electrolyte were determined on 15 and 30 days of the administration; Urine albumin (mAlb), kidney injury molecule-1 (KIM-1), neutrophil gelatinase-associated lipocalin( NGAL), N-acetyl-beta-D-glucosaminidase (NAG), clusterin, beta2-microglobulin (beta2-MG), alpha1-microglobulin (alpha1-MG), alanine aminopeptidase( AAP) and im- munoglobulins IgG were tested on 15 and 30 days of the administration.

RESULTCompared with the control group, urine protein and white blood cells was significantly increased in each dose group. On 15 days of the administration, mAlb were higher in each dose group, KIM-1, NGAL, clusterin, NAG and AAP were significantly higher in high-dose group, while the middle and low dose group had no significant difference, as well as blood SCr and BUN no obvious abnormalities. On 30 days, mAlb, KIM-1, clusterin, NAG, AAP were increased in each dose group, appearing dose-effect relationship, beta2-MG and NGAL levels were significantly increased in high-dose group. Contents above indicators were increased with significant dose and time relationship, and serum BUN, Scr were correlated, suggesting that urine mAlb, KIM-1, clusterin, NAG and AAP indicators that can sensitively respond the changes of proteins and enzymes in urine.

CONCLUSIONMethyl cantharidimide tablets has a renal toxicity, urine mAlb, KIM-1, clusterin, NAG and AAP can be used as the early nephrotoxic biomarkers of methyl cantharidimide tablets.

Animals ; Biomarkers ; urine ; Dogs ; Female ; Kidney ; drug effects ; Kidney Diseases ; chemically induced ; Male ; Proteins ; metabolism ; Tablets ; adverse effects ; Urine ; chemistry

Adult ; Aged ; Humans ; Male ; Peliosis Hepatis

Objective To determine the impact of uric acid changes on kidney function in middle-aged and elderly women with normal renal function.Methods Serum uric acid and estimated glomerular filtration rate(eGFR) were studied by prospective cohort analysis in 1152 middle-aged and elderly Dalian women with serum uric acid<7 mg/dl, estimated glomerular filtration rate(eGFR)≥90 ml·min-1·(1.73 m2)-1, and age>45 years old.Results The analysis of variance (ANOVA) of clinical indices between baseline and follow-up data showed that an eGFR was decreased after three years in both middle-aged and elderly women even with normal serum uric acid.Compared with those in the lowest guartile subgroup, a significant drop of eGFR to less than 90 ml·min-1·(1.73 m2)-1 was seen in those with the highest guartile of baseline serum uric acid group (OR=1.707, 95%CI 1.192-2.443, P=0.003).Furthermore, there were positive correlations between changes in the levels of serum uric acid and eGFR, and linear regression analysis showed that, as the change level of serum uric acid increased, the change level of eGFR declined(β=1.562, 95%CI 1.050-2.075, P<0.01], and the changing degree of uric acid increase is independent risk factors refor eGFR reductions.Conclusion Reduction in eGFR level is inversely related to serum uric acid in perimenopausal and postmenopausal women.However, our study showed that, women with serum uric acid level ranging from normal, slight increase, and relatively high levels, the increasing degrees of uric acid are independent risk factors responsible for the changing degree of eGFR reduction.

Adult ; Aged ; Blood Pressure ; physiology ; Earthquakes ; Female ; Heart Rate ; physiology ; Humans ; Male ; Middle Aged

OBJECTIVETo investigate into the signaling pathway of Porphyromonas gingivalis (P. gingivalis) on cytokine expression in human dental pulp cells (HDPC).

METHODSAnaerobic method was employed to culture P. gingivalis, and then HDPC were intracellularly infected by P. gingivalis. The extraction of total RNA, real-time quantitative polymerase chain reaction (qPCR) and enzyme linked immunosorbent assay (ELISA) was used for mRNA expression of Nods and Rip2, protein secretion of interleukin-6 (IL-6).

RESULTSHDPC expressed Nods, Rip2 mRNA and IL-6. The up-regulation of Nods and Rip2 mRNA started after P. gingivalis infection, reached maximal level at 2 h, and then decreased at 6 h; whereas elevated IL-6 was found when P. gingivalis infected.

CONCLUSIONP. gingivalis activate host innate immune responses in HDPC, and induce IL-6 production through Nod/Rip2-mediated signaling pathway.

Cells, Cultured ; Humans ; Interleukin-6 ; Interleukin-8 ; Porphyromonas gingivalis ; RNA, Messenger ; Up-Regulation

OBJECTIVETo investigate the relationship between the helicobacter pylori (HP) infection and the genetic instability of mitochondrial DNA (mtDNA) in human gastric adenocarcinoma epithelial cells (AGS).

METHODSAfter treated with extracts of HP11638 (CagA+, VacA+) or Hp11638 mutant strain (CagA+, VacA-), AGS cells were collected, and mitochondrial DNA was extracted and Cox-I, Cox-II, Cox-III, ATPase6, ATPase8 and Cytb genes and the D-Loop region were amplified by PCR and then sequenced.

RESULTSThe mutation rates of the mtDNA in AGS cells were correlated with the extracts of the two HP strains in a concentration- and time-dependent manner. But the mtDNA mutation rate in AGS cells treated with the HP11638 extract was higher than that treated with the Hp11638 mutant extract. Total of 616 mutations in D-Loop region were detected, including 489 point mutations, 81 insertions and 46 deletions. Among them, 70.9% (437/616) belonged to GC to AT and AT to GC transition. Seventeen out of 20 (85%) AGS cells treated with extract of HP had mutations in 303PolyC, 16184PolyC and 514CA regions of mtDNA D-Loop. No mutation was detected in Cox-I, Cox-II, Cox-III, ATPase6 and ATPase8 genes, three point mutations were found in the Cytb gene.

CONCLUSIONHP can cause the accumulation of mutations in mtDNA, in particular, in the D-Loop region, and the VacA participated in the process.

Antigens, Bacterial ; pharmacology ; Base Sequence ; DNA, Mitochondrial ; drug effects ; genetics ; Endothelial Cells ; drug effects ; pathology ; Helicobacter Infections ; complications ; Helicobacter pylori ; chemistry ; Humans ; Mutation ; Stomach ; pathology

OBJECTIVETo investigate whether simian virus 40 (SV40) was related to patients of malignant mesothelioma in China.

METHODSParaffin-embeded samples of 17 patients with malignant mesothelioma were collected. After isolation of DNA from paraffin blocks, polymerase chain reaction (PCR) analyses were performed using three different sets of primer for detection of SV40 large T antigen gene. These samples were also immunohistochemically evaluated for expression of SV40 TAg protein with two different anti-SV40 Tag (Pab101 and Ab-2).

RESULTSOnly one of the three primer pairs successfully amplified SV40 genome in three malignant mesothelioma samples. No immunopositive staining for SV40 TAg was found in any of the samples.

CONCLUSIONSThe study shows that malignant mesothelioma in China may be independent of SV40 infection.

Adult ; Aged ; Antigens, Viral, Tumor ; genetics ; metabolism ; China ; Female ; Host-Pathogen Interactions ; Humans ; Immunohistochemistry ; Male ; Mesothelioma ; pathology ; virology ; Middle Aged ; Polymerase Chain Reaction ; Polyomavirus Infections ; pathology ; virology ; Simian virus 40 ; genetics ; immunology ; physiology ; Tumor Virus Infections ; pathology ; virology ; Young Adult

OBJECTIVETo investigate the outcome of the children with thoracic spine tuberculosis who underwent radical debridement, reconstruction with bone autograft or allograft and internal fixation via posterior approach.

METHODSFrom June 2005 to December 2010,9 children with thoracic spine tuberculosis underwent radical debridement, reconstruction with bone autograft or allograft and internal fixation via posterior approach including 7 males and 2 females with an average age of 7 years old ranging from 3 to 12 years. The course of the disease ranged from 3 months to 1 year (averaged 6 months). All the patients had continuous thoracic and back pain,intercostal nerve pain and kyphosis deformity, accompanied with low fever,night sweat and pathologic leanness. The predilection sites examined by X-ray, CT or MRI were in T4 to T9 segment. The kyphosis angle ranged from 35 degrees to 72 degrees (averaged 48.2 degrees) before surgery. The ASIA classification was as follows: 2 cases at grade B, 5 at grade C, 2 at grade D. All the patients underwent a standard one-stage operation via posterior approach. Radical debridement was performed, then iliac crest bone autograft or allograft was placed and transpedicular screw system internal fixation was done to reconstruct the spinal column. The change of kyphosis angle and fusion of bone grafting were reexamined by X-ray regularly. The neurological function were evaluated according to ASIA classification.

RESULTSThere was no injury of blood vessel or spinal cord during the surgery. Nine children were followed-up for 16 to 38 months (averaged 24 months). The tuberculosis symptoms disappeared after surgery and there was no tuberculosis recurrence,incision infection, sinus formation and internal fixation failure in any of these chiildren. ESR reexamination recovered normally. Bony fusion was obtained in all patients and internal fixation position was normal 4 to 8 month postoperatively. The kyphosis angle ranged from 12 degrees to 30 degrees (averaged 19.50) at final followed-up. The function of spinal cord improved postoperatively, the function of spinal cord recovered at different degrees: 2 cases at grade C, 2 at grade D, and 5 at grade E.

CONCLUSIONThe one-stage posterior approach can provide direct and safe access to the lesion. The effect of vertebral canal decompression and kyphosis deformity correction were significantly. The structural iliac crest autograft or allograft and posterior transpedicular screw system could work effectively to stabilize the thoracic junction.

Bone Transplantation ; Child ; Child, Preschool ; Debridement ; methods ; Female ; Fracture Fixation, Internal ; methods ; Humans ; Male ; Thoracic Vertebrae ; surgery ; Tuberculosis, Spinal ; surgery