3.To screen for SQSTM1/p62 gene in Chinese patients with familial amyotrophic lateral sclerosis carrying superoxide dismutase 1 mutation
Chinese Journal of Internal Medicine 2014;53(12):957-960
Objective To explore the clinical features and SQSTM1/p62 gene mutations in Chinese Han patients with familial amyotrophic lateral sclerosis linked superoxide dismutase 1 (SOD1) mutation (FALS-SOD1).Methods A total of 13 FALS-SOD1 probands and 100 healthy controls were studied,with DNA extracted from the peripheral blood.Sequencing was carried out at 8 exons,intron-exon boundaries and promoter region (2-kb upstream from the coding sequence) of SQSM1/p62.Clinical data were collected and all patients were followed-up.Phenotype-genotype relationship was analyzed.Results The insertion of T was found in intron 5 of SQSTM1/p62 gene [+ 1 insert T (TT > TG)] in a FALS-SOD1 G16A male proband,with limbs as the symptom onset and faster disease progression than the other two SOD1 G16A probands without SQSTM1/p62 gene mutation.Conclusions The insertion of T in the intron 5 of SQSTM1/ p62 gene may promote the ALS progression by damaging p62 function in the FALS-SOD1 G16A proband.
5.Expression of microRNA-29 b in mice with amyotrophic lateral sclerosis
Yi YANG ; Bin CAI ; Dongsheng FAN
Journal of Peking University(Health Sciences) 2015;(5):733-736
Objective:To investigate microRNA-29b ( miR-29b) expression in cerebral cortex , spinal cord, fore limb muscle, and serum of SOD1-G93A amyotrophic lateral sclerosis ( ALS) mice, and to identify the biomarker and to assess diagnostic values for ALS .Methods:Cerebral cortex , spinal cord , fore limb muscle and serum from 16 SOD1-G93 A ALS mice and 16 wild-type mice were taken and then microRNA extracted , detecting the expression of miR-29 b by real-time quantitative polymerase chain re-action ( RT-qPCR ) .The diagnostic performance of miR-29b for ALS was estimated by the receiver operating characteristic ( ROC ) curve . Results: The results from the validation indicated that the differences in miR-29b between the cerebral cortex of SOD1-G93A ALS and the healthy control subjects were statistically significant (P=0.001).Meanwhile, the expressions 8, 12, and 16 weeks later were higher than those of the controls ( ALS vs.Control: 8 weeks, P=0.044; 12 weeks, P=0.018; 16 weeks, P=0.045).When the relative expression level of miR-29b was used to diagnose ALS in SOD1-G93A ALS mice, the area under the ROC (area under the curve, AUC) was 0.885, if the diagnostic threshold was set at 0.185 6, the sensitivity and specificity were 92.9%and 71.4%.Conclusion:MiR-29 b may act as medical monitoring indices of ALS in early time .
6.Low Field MRI Diagnosis of Adenomyosis
Zisheng YI ; Yiping LIU ; Fan YANG
Journal of Practical Radiology 1992;0(11):-
Objective To investigate the diagnostic value of low field MRI for adenomyosis. Methods MRI features of adenomyosis pathologically proved in 18 cases were retrospectively analysed.Results 15 cases were diffusive adenomyosis,the junctional zones of uterus were exteusive thickened to 12.0~32.6 mm,mean 16.2 mm,diffusive high signal intensity distributed over in myometrium which was low signal intensity in 11 cases,it was typical “snowing sign”,lower signal intensity in the myometrium in another 4 cases on T 2WI and fat-suppression imaging. A little high signal intensity was found in 6 cases on T 1WI. 3 cases were focal adenomyosis(adenomyoma), 4 lesions totally. The adenomyoma’s boundaries were not distinct and their shapes were roundish or irregular. The lesions were low signal intensity or diffusive high signal intensity distributed in the low signal intensity fields on T 2WI and fat-suppression imaging. A little high signal intensity was found in 2 lesions on T 1WI. Conclusion T 2WI and fat-suppression imaging of low field MRI are very useful techniques of the diagnosis of adenomyosis.
7.Study of 34 cases with lung nodules SPECT-CT imaging with 99m Tc-MIBI
Lei YANG ; Weihong YUAN ; Yi ZHANG ; Jianxian YANG ; Fan YANG
Journal of Chinese Physician 2011;(z1):1-3
Objective To evaluate the value of the fusion of 99m Tc-MIBI imaging technology apply in the diagnosis of malignant lung tumor. Methods Thirty - four cases with lung nodules proved by X-ray and/or CT scanning, a total of 48 lung nodular lesions. And the imaging with-99m Tc-MIBI of chest performed at 10 minutes and 2 hours delayed after injection by GE Infinia Hawkeye 4 SPECT-CT. then the regions of interesting ( ROI) were drawn in the tumor and contra lateral position to calculate the radioactivity ratios of tumor to normal ( T/NT) , and fused with the spiral CT scanning image in the same machine, and reading the early and delayed image respectively. Judged the result of the image develops, and statistical analysis of the ratio (T/NT) according to the final pathologic consequence. Results All cases with total of 48 nodular lesions, 21 nodules were positive in early imaging, 16 nodules were positive in delayed imaging (the ratio T/NT over 3. 33). defined the delayed image positive as the final criterion, The(T/NT)ratios of Malignant lung lesions were significantly higher than the benign lesions ( P <0. 05). Negative nodes 27, 13 cases of lung cancer lesions were malignant, confirmed by postoperative pathologic examination. The falsepositive nodules 3, false-negative nodules 2. The sensitivity was: 88.88%, the specificity was: 90.9% positive predictive value ( +PV) was: 84. 21% , negative predictive value (-PV) is: 93.75%. Conclusion 99mTc-MIBI as a tumor positive imaging agent is highly sensitivity to lung lesions, but specificity is not so high.
8.Relation between drug release and the drug status within curcumin-loaded microsphere.
De CHEN ; Yi LIU ; Kai-yan FAN ; Yi-qiao XIE ; An-an YU ; Zi-hua XIA ; Fan YANG
Acta Pharmaceutica Sinica 2016;51(1):140-146
To study the relation between drug release and the drug status within curcumin-loaded microsphere, SPG (shirasu porous glass) membrane emulsification was used to prepare the curcumin-PLGA (polylactic-co-glycolic acid) microspheres with three levels of drug loading respectively, and the in vitro release was studied with high-performance liquid chromatography (HPLC). The morphology of microspheres was observed with scanning electron microscopy (SEM), and the drug status was studied with X-ray diffraction (XRD), differential scanning calorimetry (DSC) and infrared analysis (IR). The drug loading of microspheres was (5.85 ± 0.21)%, (11.71 ± 0.39)%, (15.41 ± 0.40)%, respectively. No chemical connection was found between curcumin and PLGA. According to the results of XRD, curcumin dispersed in PLGA as amorphous form within the microspheres of the lowest drug loading, while (2.12 ± 0.64)% and (5.66 ± 0.07)% curcumin crystals was detected in the other two kinds of microspheres, respectively, indicating that the drug status was different within three kinds of microspheres. In the data analysis, we found that PLGA had a limited capacity of dissolving curcumin. When the drug loading exceeded the limit, the excess curcumin would exist in the form of crystals in microspheres independently. Meanwhile, this factor contributes to the difference in drug release behavior of the three groups of microspheres.
Calorimetry, Differential Scanning
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Curcumin
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chemistry
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Drug Liberation
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Lactic Acid
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Microscopy, Electron, Scanning
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Microspheres
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Polyglycolic Acid
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X-Ray Diffraction
9.Impact of Xuebijing injection on the expression of tumor necrosis factor α and high mobility group box 1 in rat peritoneal mesotheliai cells induced by lipopolysaccharide
Shuo CHEN ; Yi FAN ; Jianfei MA ; Lina YANG ; Xiuli ZHANG
Chinese Journal of Nephrology 2010;26(1):34-38
Objective To observe the effect of Xuebijing injection on the expression of tumor necrosis factor-alpha (TNF-α) and high mobility group box-1 protein (HMGB-1) in rat peritoneal mesothelial cells (PMCs) induced by lipopolysaccharide (LPS). Methods PMCs were isolated from rat colic omentum and the 3rd generation cells were used in the experiment. PMCs were incubated with LPS at different concentrations (1,10,100 mg/L);with LPS (10 mg/L) for 2, 6, 12, 18, 21, 24, 36 h;with Xuebijing injection at different concentrations (2,10,20 g/L) after incubation with LPS (10 mg/L) for 2 h. PMCs in the control group were incubated with medium. HMGB-1 mRNA was detected by RT-PCR. TNF-α and HMGB-1 protein in supernatants was detected by ELISA. Results Compared to the control group, the expression of HMGB-1 mRNA and protein was significantly increased in groups stimulated by LPS in a time- and dose-dependent manner (all P<0.05);the expression of TNF-α was increased in the groups stimulated by LPS in a dose-dependent manner (P<0.05). In the groups stimulated by LPS (10 mg/L), the expression of TNF-α appeared double hump within 36 hours. Compared to LPS (10 mg/L) group, Xuebijing injection significantly inhibited the expression of HMGB-1 and TNF-α (all P<0.05 ) in a dose-dependent manner. Conclusions HMGB-1 as a late mediator of inflammatory responses may play a role in the pathogenesis of peritoneal dialysis related peritonitis. Xuebijing injection can reduce peritoneal inflammatory impairment by inhibiting the up-regulation of TNF-α and HMGB-1 induced by LPS.
10.Protein expression profile of human glomerular mesangial cells under high glucose
Shuyan DU ; Qiuling FAN ; Lining WANG ; Gang YANG ; Yi JIANG
Chinese Journal of Nephrology 2010;26(9):671-677
Objective To analyze the protein expression profile of human glomerular mesangial cells (HMCs) under high glucose and to characterize molecular functions and biological processes. Methods HMCs were divided into high glucose cultured group (30 mmol/L) and normal glucose cultured group (5 mmol/L). The total proteins were extracted after culture for 48 hours. The total proteins of the two groups were separated using two-dimensional fluorescence difference in gel electrophoresis (2-D DIGE) and analyzed using DeCyder 2-D difference analysis software. The differentially expressed proteins were further identified using in-gel digestion with trypsin, of which peptide extracts were prepared for MALDI-TOF-MS analysis. Protein identifications were searched in the NCBI protein database using the Mascot search engine. Results One hundred and forty-seven protein spots whose expression levels were significantly increased or decreased more than 1.5 folds under high glucose were identified. Ninety-six differentially expression protein spots were analyzed by peptide mass fingerprinting and 37 kinds of proteins were identified. The protein spots of phosphatidylethanolamine binding protein 1 (PEBP-1), granulysin,ATP synthase H + transporting mitochondrial FO complex subunit F2 were observed only in high glucose group. The expression of 24 proteins was up-regulated by high glucose, including eosinophil cationic protein, RGS membrane-interacting proteins 16 (MIR16), peptidyl-prolyl cis-trans isomerase, disks large homolog DLG2, breast cancer 2, early onset (BRCA2), Catechol-O-methyltransferase etc. The expression of 5 proteins was down-regulated by high glucose, including O-GlcNAc transferase-interacting protein 106 000 isoform 1, proteasome beta 6 subunit precursor,NEFA-interacting nuclear protein NIP30 etc. Conclusions Expression of 147 proteins in HMCs alters under high glucose. These proteins are involved in the regulation of cytoskeleton, glucose metabolism, cell division, gene transcription, signal transduction, phosphorylation, cell proliferation,apoptosis etc. In-depth analysis of these differentially expressed proteins' function and crosstalk is expected to provide an important experimental basis for clarifying the pathogenesis of diabetic nephropathy.