AIM: To investigate the protective effect of microRNA-218 (miR-218) silencing on kidney tissue of streptozotocin (STZ)-induced diabetic nephropathy rats and the potential mechanism.METHODS: The diabetic rat model was established by a single intraperitoneal injection of STZ (50 mg/kg).Meanwhile, the miR-218 short hairpin RNA (shRNA) lentiviral vector was constructed.The Sprague-Dawley rats were randomly divided into 4 groups: healthy control group, diabetes group, empty vector group and miR-218-shRNA group.The blood glucose, 24 h urinary protein, serum creatinine (SCr) and blood urea nitrogen (BUN) in the rats at different time points (4, 8 and 12 weeks) were measured by an automated analyzer.The expression of miR-218 was detected by RT-qPCR, while the expression of heme oxygenase-1 (HO-1), nephrin and p38 mitogen-activated protein kinase (p38 MAPK) at mRNA and protein levels in the kidney tissues was determined by RT-qPCR and Western blot.The caspase-3 activity was detected by caspase-3 activity assay kit, and the cell apoptosis of the kidney tissues was analyzed by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL).RESULTS: Compared with healthy control group, the expression of miR-218 was significantly increased in STZ-treated rats.Meanwhile, the concentrations of blood glucose, 24 h urinary protein, SCr and BUN were significantly increased in STZ-treated rats (P<0.05).The mRNA and protein expression of HO-1 and nephrin was significantly decreased, while the level of phosphorylated p38 MAPK was significantly increased in STZ-treated rats.In addition, the activity of caspase-3 was also significantly increased in STZ-treated rats.When the model rats were infected with miR-218-shRNA, the expression of miR-218 was significantly decreased and the above effects were markedly reversed.Furthermore, TUNEL results showed that compared with diabetic group and empty vector group, miR-218 silencing significantly attenuated the cell apoptosis in the kidney tissues in miR-218-shRNA group.CONCLUSION: miR-218 is involved in the kidney injury in diabetic rats, and silencing of miR-218 by lentiviral vector-mediated miR-218-shRNA transfection effectively inhibits kidney cell apoptosis, suggesting that miR-218 is a potential target for the treatment of diabetic nephropathy.

This study is to explore the interventional effects of fluvastatin on anti-beta2GPI/beta2GPI-induced activation in THP-1 mononuclear cells. In vitro, human mononuclear cells THP-1 were treated with fluvastatin, LPS and anti-beta2GPI/beta2GPI, then the TF expression on THP-1 cells was detected by real-time quantitative PCR (RT-qPCR) or TF activity was detected by kit. TNF-alpha mRNA and its protein expression were investigated by RT-PCR and ELISA kit. The expression of phospho-NF-kappaB p65 and inhibitory protein of NF-kappaB (IkappaB-alpha) were measured by Western blotting. The results suggested that the expression of TF and TNF-alpha on THP-1 cells was significantly up-regulated with treatment of anti-beta2GPI/beta2GPI complex (100 mg x L(-1)), compared with that of untreated cells (P < 0.05). Fluvastatin (50 mg x L(-1)) could decrease TF (mRNA and activity) expression and the level of TNF-alpha (mRNA and protein) in THP-1 cells with anti-beta2GPI/beta2GPI complex. The expression of TF and TNF-alpha was shown in a concentration-dependent manner. Moreover, anti-beta2GPI/beta2GPI complex could downregulate IkappaB-alpha levels and increase the levels of phospho-NF-kappaB p65. And these effects of anti-beta2GPI/beta2GPI complex could be blocked by fluvastatin. In conclusion, fluvastatin may interfere the expression and regulation of NF-kappaB signal transduction pathway, thereby inhibit the effects of anti-beta2GPI/beta2GPI on activation of THP-1 cells, by decreasing the expression of TF and TNF-alpha.

Virtual slides were applied in experiment teaching of pathology on trial in clinic medicine college of Sichuan university.The resuhs from the Survey showed that students and teachers preferred virtual slides in learning microscopic lesions.Virtual Slides can help us save time,promote quality of observation,carry out discussion based teaching and manage teaching documents.It can be used in network teaching after marking the lesions.

We reported a case of pulmonary capillary hemangiomatosis (PCH) and introduced its diag-nosis, differential diagnosis, pathogenesis and development of treatment based on the review of Dana Point 2008 Classification of Pulmonary Hypertensiona and current literatures .A 43-year-old female presented progressive dyspnea, elevated pulmonary arterial pressures and CT pulmonary angiography (CTPA) imaging of main pulmonary arterial enlargement and wide spread ill -defined centrilobular nodules of ground-glass opacity.Her histologic features were proliferation of capillary channels within alveolar walls as well as muscularization of arterioles and medial hypertrophy of muscular pulmonary arteries.The treatment with diuretics and warfarin was used promptly , but unfortunately was ineffective. The patient died three months after diagnosis .PCH is a very rare vascular disease with poor prognosis . The diagnosis of PCH rests on the integration of clinical and radiographic information with pathologic fea -tures, however pathology is the most reliable means .Because clinical symptoms, imaging and histological features of pulmonary veno-occlusive disease (PVOD) and PCH broadly overlap, differential diagnosis should be made carefully.Among the various pathologic features proliferation of capillaries within alveolar walls is the key point for diagnosing PCH , which is also the most critical criteria for differentiating PCH from PVOD.So far the only definitive treatment for PCH is lung transplantation , without which the pa-tient will die several months after diagnosis .

Tilianin was separated and authenticated from the seeds of Dracocephalum moldavia, a Uygur medicine, by chromatographic technique and spectroscopic method. The purity of tilianin is more than 98% determined by HPLC area normalization method. Thin layer chromatography (TLC) method was used to separate tilianin from D. moldavia by mixture of chloroform-methanol (5: 1) as a developing solvent on high performance silicagel precoated plate (SGF254) and using aluminium trichloride as a chromogenic agent for qualitative identification of D. moldavia. To establish a HPLC method for quantitative analysis of D. moldavia, tilianin was used as a Quantitative marker and separated on a C18 (4.6 mm x 250 mm, 5 &mu;m) column with acetonitrile-01% formic acid (25: 75) as the mobile phase and detected at 330 nm. The calibration curve of tilianin displayed ideal linearity over the range of 0.617 2-123.44 &mu;g x mL(-1) with a regression equation of Y = 33.773X - 0.824 8 (r = 1). The average recovery of tilianin was 101.0% with RSD of 3.7%. The RSD values of intra-day and inter-day precision were less than 2%. The content of tilianin in 4 batches of the authenticated semen of D. Moldavia was between 0.016 and 0.187 mg x g(-1). The qualitative and quantitative method established is suitable for the quality evaluation and assessment of semen of D. Moldavia.
Chromatography, High Pressure Liquid ; Chromatography, Thin Layer ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; standards ; Flavonoids ; chemistry ; isolation & purification ; standards ; Glycosides ; chemistry ; isolation & purification ; standards ; Lamiaceae ; chemistry ; Magnetic Resonance Spectroscopy ; Quality Control

OBJECTIVETo investigate the role of 83 site in interaction of GluR2 C-terminal and PICK1 PDZ domain.

METHODSDocking structure of PICK1 PDZ domain with GluR2 C terminal PDZ binding motif was built with computer software. After K83 site was substituted by other amino acid, the structure and binding energy were recalculated; meanwhile, site specific mutants were constructed using wild type full length cDNA as template. Mutants were co-transfected with GluR2 into HEK293T cells. After staining, the distribution of PICK1 and GluR2 were observed under confocal microscope.

RESULTSWild type PICK1 and GluR2 formed many co-clusters in HEK293T cells as reported by other research groups; but different K83 mutant had different distribution in HEK293T cells.

CONCLUSIONThe K83 site in PDZ domain of PICK1 is important for the interaction between PICK1 and GluR2. Altering lysine will probably change the hydrophobic interactions, the hydrogen bonds or the electrostatic interactions formed between PICK1 PDZ domain and GluR2 C terminal; accordingly, that will change the binding capacity between PICK1 and GluR2 in varying degrees.

Binding Sites ; Carrier Proteins ; chemistry ; metabolism ; Computer Simulation ; HEK293 Cells ; Humans ; Nuclear Proteins ; chemistry ; metabolism ; PDZ Domains ; Protein Binding ; Receptors, AMPA ; metabolism

OBJECTIVETo construct a method for detecting the copy number of survival of motor neuron 1 gene (SMN1) with single copy difference based on real-time fluorescence quantitative PCR, and to make practical use of the method for acquiring the data on SMN1 copy number in Chinese as well as for screening the carriers of spinal muscular atrophy (SMA) from healthy individuals and SMA families.

METHODSExon 7 and flanking area of SMN1 gene were amplified by real-time fluorescence quantitative PCR in 264 healthy individuals, in 1 standard sample having 2 SMN1 but having no SMN2, and in 88 parents of SMA patients. The samples for detecting were diluted to 30 ng/microL and the standard sample was diluted to 15 ng/microL, 30 ng/microL, 45 ng/microL, 60 ng/microL; the unknown samples and 4 standard samples with different concentrations were amplified at the same time, a standard curve could be drawn out according to the results of the 4 standard samples, then the copy number of samples could be calculated.

RESULTSOf 88 parents' samples, 84 samples each had 1 copy of SMN1, and the rest 4 each had 2 copies of SMN1. Of 264 healthy individuals' samples, 5 samples each had only 1 copy of SMN1 (an indicator of definite gene carriers), 232 samples each had 2 copies of SMN1, 25 samples each had 3 copies of SMN1, and 2 samples each had 4 copies of SMN1. Of the samples of 32 members of SMA families, 2 samples each had only 1 copy of SMN1 indicating definite gene carriers, 25 samples each had 2 copies of SMN1, and 5 samples each had 3 copies of SMN1.

CONCLUSIONSMN1 copy number could be detected precisely by real-time fluorescence quantitative PCR; the screening of gene carriers could provide essential data for genetic counseling.

Exons ; Family Health ; Female ; Fluorescence ; Gene Dosage ; Humans ; Male ; Muscular Atrophy, Spinal ; genetics ; Polymerase Chain Reaction ; methods ; Survival of Motor Neuron 1 Protein ; genetics

OBJECTIVETo introduce the application of denaturing high-performance liquid chromatography (DHPLC) in the diagnosis of childhood type spinal muscular atrophy (SMA).

METHODSExon 7 and flanking area of survival motor neuron (SMN) gene were amplified by PCR in 1 standard sample, 25 normal individuals and 25 patients with SMA. The PCR products were then directly loaded onto the DHPLC system after denaturing and annealing. Different DNA segments were separated by changing the concentration of buffer A relative to that of buffer B.

RESULTSDifferent DNA segments were separable on the DHPLC chromatogram. Three peaks including SMN1/SMN2 heteroduplex peak, SMN2 homoduplex peak and SMN1 homoduplex peak were detected in 23 out of 25 normal individuals. Only SMN1 homoduplex peak was detected in 2 normal individuals and the standard sample, indicating the deletion of SMN2 On the contrary, only the SMN2 homoduplex peak was detected in 22 out of 25 patients with SMA, indicating deletion of SMN1. The three peaks as those of normal individuals were detected in the other 3 patients, indicating no SMN1 or SMN2 deletion.

CONCLUSIONAs a new technology for diagnosing SMA, DHPLC is sensitive, accurate, rapid and convenient.

Chromatography, High Pressure Liquid ; methods ; Exons ; genetics ; Humans ; Muscular Atrophy, Spinal ; diagnosis ; genetics ; Polymerase Chain Reaction ; Reproducibility of Results ; SMN Complex Proteins ; genetics ; Sensitivity and Specificity ; Survival of Motor Neuron 1 Protein ; genetics ; Survival of Motor Neuron 2 Protein

OBJECTIVETo improve the cure rate of patients with abdominal visceral injury complicated by craniocerebral injury.

METHODSClinical data of 176 cases of abdominal visceral injury complicated by craniocerebral injury were retrospectively analyzed.

RESULTSIn this series, 44 cases died and the mortality was 25.0. The main cause of death is abdominal visceral injury combined with shock and severe craniocerebral injury.

CONCLUSIONSIt is essential to improve the cure rate by accurate diagnosis at early stage. Abdominal paracentesis and CT should be performed promptly and dynamically. Priority should be given to the treatment of life-threatening injuries.

Abdominal Injuries ; diagnosis ; therapy ; Adolescent ; Adult ; Aged ; Child ; Craniocerebral Trauma ; diagnosis ; therapy ; Female ; Humans ; Male ; Middle Aged ; Multiple Trauma ; diagnosis ; therapy ; Viscera ; injuries

OBJECTIVETo measure lutein, zeaxanthin and β-carotene level in foods commonly consumed in Beijing, and compare the content difference between raw and cooked food.

METHODSForty-six commonly consumed foods of 8 classes were collected in Haidian district of Beijing from September to October in 2009. A high performance liquid chromatography method was used to determine the content of lutein, zeaxanthin and β-carotene in both raw and cooked samples.

RESULTSLutein was abundant in cucurbitaceous and solanaceous, allium and nuts, especially in Chinese chive (18 226.9 µg/100 g) and pumpkin (13 265.2 µg/100 g). Major sources of zeaxanthin included round pumpkin, green garlic shoot, corn and eggs, whose level of zeaxanthin were 444.6, 283.5, 279.7, 118.6 - 377.9 µg/100 g, respectively. Zeaxanthin level of those cooked foods changed to 483.9, 239.3, 279.1, 149.5 - 594.7 µg/100 g, respectively. The zeaxanthin level of cooked Chinese chive reached 1081.2 µg/100 g, while we did not detect any zeaxanthin in raw Chinese chive. β-carotene was present in a wide variety of vegetables and fruits. Carrot (17 234.3 µg/100 g) was a good source of β-carotene, while its level in cooked carrot was 17 013.5 µg/100 g.

CONCLUSIONConsuming the proper kinds of foods and changing the method of food processing were beneficial to increase the intake of lutein, zeaxanthin and β-carotene.

China ; Cooking ; Food ; Food Analysis ; Lutein ; analysis ; Xanthophylls ; analysis ; Zeaxanthins ; beta Carotene ; analysis