Objective:To construct NBS1 microRNA expressing eukaryotic recombinants,and identify biological activity of recombinants in Hela cell after transfection.Methods:According to sequence of NBS1mRNA,the NBS1 pre-microRNA was designed and synthesized,then cloned into the GFP reporter pcDNA6.2-GW/EmGFP-miR vector and transfected into Hela cell line.To detect integrity of inset fragment through colony PCR and sequencing analysis.The biological activity of recombinants through identify interference efficiency of NBS1 microRNA recombinants by way of Real-Time PCR and Western blot were determined.Results:Sequences of inset fragment in four microRNA expressing recombinants were correct.NBS1 mRNA and protein expression of four microRNA recombinants were decrease,which is the lowest in the NBS1mi-2 group.Conclusion:Four NBS1-targeting microRNA expressing recombinants all have biological activity in Hela cell line,and NBS1mi-2 recombinant has the most interference efficiency.The microRNA expressing plasmid which were successfully constructed and lay foundation for the studies on the tumor gene therapy of microrna targeting NBS1.

Chronic Disease ; Electric Stimulation Therapy ; Heart Failure ; therapy ; Humans ; Myocardial Contraction

0.05).Conclusion Tripterygium wilfordii,combined with routine treatment,appeared to decrease 24-hour proteinuria in a certain extent and did not adversely affect liver function,renal function and the blood routine test in most patients with diabetic nephropathy.

OBJECTIVETo observe the clinical manifestations of bullous retinal detachment and analyze the etiological factors.

METHODSA retrospective analysis of the clinical data was conducted in 22 patients with multifocal retinal pigment epitheliopathy (DRPE) and big bullous retinal detachment (BBRD), who were admitted between 2003 and 2008 in Zhongshan Ophthalmic Center with the diagnoses established by ocular fundus examination, fundus fluorescein angiography (FFA) and/or indocyanine green angiography (ICGA).

RESULTSThe patients included 15 men (68%) and 7 women (32%), with a mean age at the initial visit of 42 years, ranging from 25 to 64 years. Four patients (18%) received previously systemic corticosteroid therapy, and 2 of them used corticosteroids before retina detachment, 1 suffered progression of retinal detachment after corticosteroid therapy, and the other developed retinal detachment in the healthy eye during the therapy. Multifocal bullous retinal detachment was diagnosed as diffuse pigment epitheliopathy (DRPE) in 9 cases. Most of the 13 cases of big bullous retinal detachment had poor vision after operation and laser therapy.

CONCLUSIONBullous retinal detachment occurs most frequently in mid-life and more often in men than women. Abnormal retinal pigment epithelium (RPE) and hyperpermeability of the choroid vessels are associated with its occurrence. Systemic corticosteroid therapy and mental stress may induce and aggravate this disease. Early medication and laser therapy are effective, and surgical intervention may save only part of the vision in advanced cases.

Adult ; Female ; Fundus Oculi ; Humans ; Male ; Middle Aged ; Retinal Detachment ; diagnosis ; etiology ; pathology ; therapy ; Retrospective Studies

One hundred and thirteen newly-diagnosed type 2 diabetic patients were treated with rosiglitazone for 16 weeks. Glucose tolerance was restored in 46 cases ( group A), but not restored in 67 cases (group B). Compared with group B,the patients of group A were younger and had shorter course of diabetes and lower body mass index (BMI,P＜0.05); Homeostasis model assessment for β-cell function (HOMA-β) and earlyphase insulin secretion in group A were better than those of group B before and after treatment respectively (P＜0. 05 ) ,while homeostasis model assessment for insulin resistance ( HOMA-IR )in group A was lower than that of group B before treatment (P＜0. 05 ). In conclusions, rosiglitazone improved insulin resistance of type 2 diabetes patients, thus facilitated restoration of β-cells function and glucose tolerance. The related factors of glucose tolerance restoration included age ,diabetes duration, BMI, HbA1 c, β-cells function, and insulin resistance.

OBJECTIVE: To examine various pneumatized extensions of the sphenoid sinus of Chinese people. METHOD: The sphenoid sinus and its surrounding structures were examined from 100 computed tomography images of the sinus. The type of the sphenoid sinus was classified according to the various extensions of the sinus. RESULT: The type of the sphenoid sinus was classified into the following 6 basic types based on the direction of pneumatization: sphenoid body, lateral, clival, lesser wing, anterior, and combined. CONCLUSION The variations in the extensions of pneumatization of the sphenoid sinus may facilitate entry into areas bordering the sphenoid sinus.
Adult ; Asian Continental Ancestry Group ; Humans ; Sphenoid Bone ; anatomy & histology ; Sphenoid Sinus ; anatomy & histology ; Tomography, X-Ray Computed

Nanoporous ZnO was used as a carrier to prepare drug solid dispersion, the mechanism of which to improve the drug dissolution was also studied. Nanoporous ZnO, obtained through chemical deposition method, was used as a carrier to prepare indomethacin and cilostazol solid dispersions by melt-quenching method, separately. The results of scanning electron microscope, surface area analyzer, fourier transform infra-red spectroscopy, differential scanning calorimeter and X-ray diffraction showed that drugs were implanted into nanopores of ZnO by physical adsorption effect and highly dispersed into nanopores of ZnO in amorphous form, moreover, these nanopores strongly inhibited amorphous recrystallization in the condition of 45 degrees C and 75% RH. In addition, the results of the dissolution tested in vitro exhibited that the accumulated dissolutions of indomethacin and cilostazol solid dispersions achieved about 90% within 5 min and approximately 80% within 30 min. It was indicated in this study that the mechanism of drug dissolution improvement was associated with the effects of nanoporous ZnO carrier on increasing drug dispersion, controlling drug in nanopores as amorphous form and inhibiting amorphous recrystallization.
Anti-Inflammatory Agents, Non-Steroidal ; administration & dosage ; chemistry ; Calorimetry, Differential Scanning ; Drug Carriers ; Indomethacin ; administration & dosage ; chemistry ; Microscopy, Electron, Scanning ; Nanostructures ; Phosphodiesterase 3 Inhibitors ; administration & dosage ; chemistry ; Solubility ; Spectroscopy, Fourier Transform Infrared ; Tetrazoles ; administration & dosage ; chemistry ; X-Ray Diffraction ; Zinc Oxide ; chemistry

OBJECTIVETo compare the transduction efficiencies of adenoviral vector, adeno-associated viral vector, baculoviral vector, and plasmid vector in human bone-marrow-derived mesenchymal stem cells (hBMSCs).

METHODSThe hBMSCs were cultured in vitro and transducted with the adenoviral vector, adeno-associated viral vector, baculoviral vector, and plasmid vector. The expression of target protein was observed by inverted fluorescent microscopy and flow cytometry.

RESULTSInverted fluorescent microscopy showed that some of the hBMSCs after transduction expressed the green fluorescent protein (GFP) and the hBMSCs transducted with baculoviral vector expressed more GFP than those of other three vectors. Flow cytometry showed that the transduction efficiencies and mean fluorescence intensities of the adenoviral vector, adeno-associated viral vector, and plasmid vector were 42%, 37%, and 22% and 158, 115, and 77, respectively, which were significantly lower than those of baculoviral vector (70%, P < 0.01; 212, P < 0.05; respectively).

CONCLUSIONCompared with the adenoviral vector, adeno-associated viral vector, and plasmid vector, the baculoviral vector has higher transduction efficiency in hBMSCs and therefore may be a more suitable gene vector for research in human gene therapy.

Adenoviridae ; genetics ; metabolism ; Baculoviridae ; genetics ; metabolism ; Bone Marrow Cells ; metabolism ; virology ; Cells, Cultured ; Dependovirus ; genetics ; metabolism ; Gene Expression ; Genetic Vectors ; genetics ; metabolism ; Green Fluorescent Proteins ; genetics ; metabolism ; Hematopoietic Stem Cells ; metabolism ; virology ; Humans ; Plasmids ; genetics ; metabolism ; Transduction, Genetic ; methods

OBJECTIVETo determine the expressions of miR-155, miR-34a and miR-30a in diffuse large B-cell lymphoma (DLBCL) and to explore their potential correlation with clinicopathological characteristics.

METHODSThe expression level of miR-155, miR-34a and miR-30a in 46 DLBCL samples were determined with TaqMan real-time polymerase chain reaction. Interphase fluorescence in situ hybridization (I-FISH) was performed to detect MYC and p53 genes' status, and immunohistochemistry (Envision method) was used to evaluate the expression of CD3, CD10, CD20, BCL-6 and MUM-1 in DLBCL. The DLBCLs were classified into germinal center B cell-like (GCB) and non germinal center B cell-like (non-GCB) subtypes according to Hans' criteria.

RESULTSCompared with normal controls, miR-155 expression level was significantly higher in DLBCL. The expression level of miR-155 in non-GCB type was higher than that in GCB type. It was shown that the patients with MYC rearrangement had lower expression level of miR-155 than the negative controls. Compared with p53 normal group, the expression level of miR-34a was significantly lower in p53 deletion group. It was also shown that the patients with BCL-6 protein expression had lower expression of miR-30a compared with the negative group.

CONCLUSIONmiR-155 expression level is different in normal controls, DLBCL and patients with subtype DLBCL. It therefore has a diagnosis value for DLBCL. miR-34a is of great prognostic significance. miR-155, miR-34a and miR-30a may be potential therapy targets for DLBCL.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Lymphoma, Large B-Cell, Diffuse ; genetics ; metabolism ; pathology ; Male ; MicroRNAs ; genetics ; metabolism ; Middle Aged ; Proto-Oncogene Proteins c-myc ; genetics ; metabolism ; Tumor Suppressor Protein p53 ; genetics ; metabolism ; Young Adult

OBJECTIVETo explore the expression of uncoupling protein-2 mRNA in brown adipose tissue, white adipose tissue and skeletal muscle of diet-induced obesity-resistant (DIO-R) rats.

METHODSFifty male Sprague-Dawley (SD) rats were randomly divided into a control group and a high-fat group and fed with basic diet and high-fat diet respectively for 13 weeks. DIO-R and DIO rats were selected according to their body weight. The change of body weight and the intake of total calorie were observed. Reverse transcription-polymerase chain reaction (RT-PCR) was used to measure the expression of UCP2 mRNA in rat.

RESULTSBody weight and total calorie intake in DIO-R rats (425.1 +/- 27.1) g, (31,693 +/- 946) kJ were significantly lower than those in DIO rats (489.7 +/- 20.5) g, (34,363 +/- 1465) kJ. The peak area of UCP2 mRNA in white adipose tissue in DIO-R rats was 352 +/- 30 and in DIO rats was 101 +/- 12. The peak areas of UCP2 mRNA in skeletal muscle in DIO-R and DIO rats were 130 +/- 15 and 170 +/- 12, respectively. The peak areas of UCP2 mRNA in brown adipose tissue of DIO and DIO-R rats were 124 +/- 14 and 147 +/- 19, respectively.

CONCLUSIONThe expression of UCP2 mRNA in white adipose tissue of DIO-R rats increased significantly. These results suggest that obesity-resistance was associated with a tissue-specific increase in UCP2 expression.

Adipose Tissue ; drug effects ; metabolism ; Animals ; Body Weight ; drug effects ; Dietary Fats ; administration & dosage ; Gene Expression ; genetics ; Ion Channels ; Male ; Membrane Transport Proteins ; genetics ; Mitochondrial Proteins ; genetics ; Muscle, Skeletal ; drug effects ; metabolism ; Obesity ; etiology ; genetics ; RNA, Messenger ; genetics ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction ; Uncoupling Protein 2