Objective To establish the method to detect the expression of apolipoprotein E gene in human adipose cells. Methods Apolipoprotein E gene expression was determined by competitive reverse transcription-PCR in adipose cells of healthy children. Results Apolipoprotein E gene expressed in the adipose cells of healthy children. The mean expression quantity of apolipoprotein E mRNA in adipose cells was (0.41?0.12 ) in healthy children. Conclusions Competitive reverse transcription-PCR is a rapid, simple, sensitive and accurate method to analyze expression of apolipoprotein E gene.

Objective To study the mechanism of calcium oxalate monohydrate and calcium oxalate dihydrate stone formation. Methods 258 urinary stones were examined by infrared spectrophotometry, from which, 20 patients with calcium oxalate monohydrate stones (COM) and 10 patients with calcium oxlate dihydrate stones (COD) were selected for the study of some urinary parameters. The statistical study was carried out with SPSS t test . Results The urinary excretions of both calcium and phosphate varied obviously between the two groups of patients.In the COM group,the urinary calcium was (4.83?1.98)mmol/24h whereas in COD group it was (9.88?4.28)mmol/24h,( P

10.3969/j.issn.2095-4344.2013.25.011

BACKGROUND:Dihydroartmisinin can promote apoptosis of glioma cels GL261, but its effect on glioma stem cels is stil unknown. OBJECTIVE:To investigate the preliminary mechanism that dihydroartemisinin inhibits migration and invasion of glioma stem cels. METHODS: Glioma stem cels were isolated from mouse malignant glioma cel lines GL261. Immunofluorescence analysis was conducted to identify the characteristics of glioma stem cels. Migration and invasion abilities of glioma stem cels were analyzed by Transwel assay. The mRNA expressions of Tol-like receptor 2, matrix metaloproteinase-2 and matrix metaloproteinase-9 were examined by real-time fluorescence quantitative PCR. RESULTS AND CONCLUSION:The characteristics of glioma stem cels were identified by CD133 and Nestin staining. The migration and invasion of glioma stem cels were attenuated by dihydroartemisinin dose-dependently. Moreover, the mRNA expression of Tol-like receptor 2, matrix metaloproteinase-2 and matrix metaloproteinase-9 was also decreased by dihydroartemisinin in a dose dependent manner. These results suggest that dihydroartemisinin inhibits the migration and invasion of glioma stem cels probably through attenuation of Tol-like receptor signaling pathway.

Objective To investigate the growth inhibition and radiosensitization of Celecoxib in hu-man nasopharyngeal carcinoma cell line CNE-2. Methods CNE-2 growth inhibition by Celecoxib was eval-uated by MTT method. Apoptosis-related changes in morphology were observed by transmission electron mi-croscopy (TEM). Cell cycle distribution and apoptosis rate were measured by flowcytometry (FCM). The ex-pression of COX-2 protein was observed by SP method after the treatment of Celecoxib. Cells were randomly planted into four groups: irradiation control(Ci), drug group(Cd), irradiation group(R), and Celecoxib plus irradiation group(D+R). Single irradiation of 2,4,6,8,and 10 Gy were administered for colonogenic assay. Cell cycle distribution and apoptosis rate were analyzed at 6 Gy irradiation. Results The growth of CNE-2 cell was inhibited by celecoxib in a dose-and time-dependent manner, the IC50 was 80 μmol/L After the treatment, cell ratio of GO and G, phases was increased (47.03±2.76 vs 56.17±1.95, t=4.68, P= 0.010), whereas the ratio of S and G2/M phases was decreased (33.07±1.86 vs 24.87±1.76, t=5.54, P = 0.010; 19.30±0.53: 17.73±0.83, t=2.75, P=0.050), and the apoptosis rate was increased (1.57±0.47:10.47±0.31, t = 27.39, P = 0.000) in a dose-dependent manner. Apoptosis with nuclear chromatin condensation, fragmentation and cell shrinkage was found by TEM. SP method showed that Celeib decreased COX-2 expression (17.48±0.34 vs 12.82±0.51,t=13.20,P =0.00). The sensitivity ratio(D0) was 1.15. FCM showed that the percentage of cells in G2/M phase was significanty more in R and D+R groups than in Ci and Cd groups (68.00±1.65,54.27±5.74,17.60±0.80,14.86±1.23, t=47.70,P=0.000; t=11.63, P=0.000), and also significantly different between R group and D + R group (t=3.99, P= 0.020). The apoptosis rate was higher in R and D + R groups than Ci and Cd groups(4.83±0.97,9.50± 1.35,1.33±0.86 and 2.28±0.42,t=4.67,P=0.010;t=8.81, P=0.000), D + R group than R group(t =4.85,P=0.010). Conclusions Celecoxib can markedly inhibit the growth and induce apoptosis in CNE-2 cells,which may depend on COX-2 pathway. Celeeoxib potently enhances the radiosensitivity of CNE-2 cells,which may due to the repair inhibit of radiation-induced DNA damage, inhibit of cell proliferation,and enhancement of cell apoptosis after irradiation.

Objective To explore the protective effects of dipeptidyl peptidase-4 inhibitor (DPP-4I) on AD-like neurodegenerative changes and its mechanism. Methods The human neuroblastoma cell line SH-SY5Y on the logarithmic phase was divided into six groups:control group (CON group, treated with PBS contained 1‰DMSO for 12 h), wortmannin intervention group (W group, treated with 0.03 μmol/L wortmannin for 12 h), DPP-4I intervention group (DPP-4I group, treated with 10μmol/L DPP-4I for 12 h), both DPP-4I and wortmannin intervention group (DPP-4I+W group, pre-treated with 10 μmol/L DPP-4I for 2 h, then 0.03 μmol/L wortmannin for 12 h), DPP-4I, wortmannin and Ex9-39 intervention group (DPP-4I+W+Ex9-39 group, pre-treated with 10μmol/L Ex9-39 for 2 h, then 10μmol/L DPP-4I for 2 h followed by 0.03μmol/L wortmannin for 12 h), and Ex9-39 intervention group (Ex9-39 group, treated with 10μmol/L Ex9-39 for 12 h). MTT assay was used to detect the cell vitality. Western blot assay was used to detect the level of total tau protein (tau-5) and phosphorylated tau at different sites (pSpS199/202, pT231 and pS396), the level of phosphorylated neurofilaments (NF-H, NF-M) and phosphorylation of critical enzyme in PI3K/Akt/GSK-3β signaling pathway. Results (1) The cell vitality decreased, the levels of pSpS199/202, pT231, pS396 and NF-H/M increased significantly in W group than those in CON group. However, comparing with CON group, the above mentioned parameters reversed in DPP-4I group. Comparing with W group, the cell vitality increased and phosphorylated levels of above mentioned indices were decreased in DPP-4I+W group. (2) The cell vitality showed a decline trend while the levels of phosphorylation tau at three different sites and NF-H/M were higher in Ex9-39 group than those in CON group. Comparing with DPP-4I+W group, the results of the phosphorylated levels showed the same changes in DPP-4I+W+Ex9-39 group. (3) Comparing with CON group, the expression levels of phosphorylated PI3K, Akt and GSK3β increased significantly in DPP-4I group, while those decreased in W group. Additionally, the expression levels of phosphorylated PI3K, Akt and GSK3β were significantly increased in DPP-4I+W group than those in W group. Conclusion DPP-4I can enhance the level of GLP-1 and activate PI3K/Akt/GSK-3βinsulin signaling pathway to improve the hyperphosphorylated tau and NFs induced by wortmannin, and to protect AD-like neurodegeneration.

Objective To evaluate the effects of chronic exposure to sub-anesthetic concentrations of sevoflurane on memory function and homeostasis of mice.Methods Thirty-six healthy male Kunming mice,aged 40 days,weighing 25-30 g,were randomly assigned into 3 groups (n =12 each):control group (group C),0.1％ sevoflurane group (group L) and 0.5 ％ sevoflurane group (group H).The mice inhaled 0.1％ (group L) or 0.5％ sevoflurane (group H) between 18:00-6:00 (the next day) every night for 30 days.Water maze test was performed at 27-30 days of inhalation.Blood samples were collected from the left ventricle for blood gas analysis and for determination of blood electrolytes.Results There was no significant difference in swimming time in Water maze test,number of errors and blood gas analysis and blood electrolytes.Conclusion Chronic exposure to subanesthetic concentrations of sevoflurane has no significant effects on memory function and homeostasis of mice.

BACKGROUND: The use of allograft tendon promotes the development of anterior cruciate ligament reconstruction, which is of great importance in the relevant basic research and clinical practice. OBJECTIVE: To summarize the current application and progress of allograft tendon in the anterior cruciate ligament reconstruction. METHODS: We searched relevant articles about the use of allograft tendon in PubMed and Embase published from January 2012 to February 2018 using the keywords of "allograft tends OR allografts, intra-articular knee ligament OR anterior cruciate ligament OR ACL". RESULTS AND CONCLUSION: China's use of allogeneic tendon in the reconstruction of the anterior cruciate ligament is still in the developmental stage. The current pros and cons of using allografts or autografts in anterior cruciate ligament reconstruction are inconclusive. Concerns about the choice of allografts are mainly due to possible infections and slow healing. A huge difference exists in different trials concerning the effects of allografts versus autografts. The conclusions are therefore roughly as follows: there is no difference in knee function after implantation of the two grafts, or the knee joint function is better after autograft. Studies have shown that allografts may increase the risk of secondary surgery. Therefore, the use of allograft tendon faces many clinical challenges in the anterior cruciate ligament reconstruction.

OBJECTIVETo explore the molecular basis for an individual with CisAB subtype of the ABO blood group.

METHODSABO antigen and serum antibody of the proband were detected with a serological method. Exons 5 to 7 of the ABO gene were amplified with PCR and sequenced bidirectionally. Allele-specific amplification for exon 6 to 7 was also carried out.

RESULTSThe proband was assigned as a CisAB phenotype based on his serological characteristics. Heterozygous variations including 220C/T, 261G/del, 297A/G, 467C/T, 646A/T, 681A/G, 771C/T, 803G/C, 829A/G and 1009A/G of the ABO gene were identified through direct sequencing, which was assigned as CisAB01var/O02 genotype. Allele-specific amplification indicated that the proband carried an O02 allele and a CisAB01var allele. Compared with A102, the CisAB01var allele has two nucleotide substitutions at 803G>C and 1009A>G, which resulted in replacement of amino acid Gly by Ala at position 268 and Arg by Gly at position 337.

CONCLUSIONThe CisAB subtype was identified with 803G>C and 1009A>G variants in the α1,3-N-acetyl-galactosaminyltransferase gene compared with that of the A102 allele.

Objective This study investigated high fat diet influence on the changes of vitamin D receptor (VDR) expression and endothelial nitric oxide synthase (eNOS) in apolipoprotein E-deficient(apoE-/-) mice.MethodsApoE-/- mice and C57BLP6J mice were divide into two groups (normal control and high fat diet),high fat diet group were feed high fat feedstuff.Plasma 25-(OH)D levels were determined by competitive protein binding radioimmunity,VDR expression were determined by immunofluorescence and reverse transcription-polymerase chain reaction.The levels of NO and eNOS were determined by nitrate reductase.ResultsCompared with normal control group,high fat diet caused more severe dam-age of atherosclerosis in wild type mice and apoE-/- mice.In apoE-/- mice,the levels of plasma 25-(OH)D were significantly decreased [(26.44±1.28) ng/mL,(22.68±2.07)ng/mL,(17.46±2.22)ng/mL,(15.88±0.97)ng/mL,P＜0.01],the expression of VDR protein and mRNA were significantly increased[VDR :0.244±0.088,0.346±0.132,0.547±0.128,0.768±0.162;VDRmRNA:0.228±0.083,0.375±0.103,0.451±0.117,0.597±0.131,P＜0.01],and the levels of NO and eNOS were significantly increased[NO:(39.74±4.81)μmol/L,(48.1±5.24 )μmol/L,(67.34±6.14 )μmol/L,(86.74±8.05)μmol/L;eNOS:(8.6±0.77 )U/L,(12.28±1.42)U/L,(15.96±0.92)U/L,(18.68±1.15)U/L,P＜0.01].These changes were more significantly in high fat diet group(P＜0.01).ConclusionsThere were abnormalities of plasma 25-(OH)D level,VDR expression and the level of NO and eNOS in apoE-/- mice.These changes were more significantly in high fat diet group.