ObjectiveTo observe the effect of digital auditory activation and touching intervention on infants with cerebral dysfunction.Methods388 infants with perinatal high-risk factors and abnormal result of Denver Development Screening Test (DDST) were randomly divided into group A (n=135), group B (n=128) and group C (n=125), and treated with digital auditory activation combined with touching (group A), simple touching (group B) and simple drug (group C) with 10 days as a course. All infants were tested with DDST after one therapeutic course, and tested again with DDST after the infants of group B and group C treated continuously for six therapeutic courses; and all infants were assessed with the Gesell development quotient (DQ) after six months.ResultsAfter one therapeutic course, normal rate of DDST was 71.11% in group A, 26.69% in group B and 20.00% in group C. After six therapeutic courses, that was 90.37 % in group A, 62.50 in group B and 40.00% in group C. After six months, the children with the Gesell DQ over 86 scores was 125 (92.60%) in group A, 90 (70.31%) in group B and 62 (49.60%) in group C. There were significant differences among three groups ( P<0.01).ConclusionThe digital auditory activation combined with touching has short therapeutic course and high efficacy, so it is a good therapeutic method for infants with cerebral dysfunction.

OBJECTIVETo explore the effects of the simultaneous activation of liver X receptor (LXR) and peroxisome proliferator-activated receptor alpha (PPARalpha) on bile acid biosynthesis in rats.

METHODSTotally 36 male SD rats were divided into three groups with 12 rats in each group: control group, high cholesterol (HC) group, and high cholesterol + fenofibrate (HC + FENO) group. Total bile acids (serum bile acids plus fecal bile acids) level was assayed. The levels of mRNA for peroxisomal palmitoyl-CoA oxidase (Acox1), LXR, cholesterol 7alpha-hydroxylase (CYP7A1), D-bifunctional protein (DBP), trihydroxycoprostanoyl-CoA oxidase (Acox2), sterol 12alpha-hydroxylase (CYP8B1), and sterol 27-hydroxylase (CYP27A1) in liver were detected by RT-PCR.

RESULTSTotal bile acid level was significantly higher in HC + FENO group than in HC group (P < 0.01), and both were significantly higher than that in control group (P < 0.01). Compared with HC group, the mRNA expression of Acox1 and DBP was significantly higher in HC + FENO group (P < 0.01), but no statistical differences was found between HC group and control group. The mRNA levels of LXR and CYP7A1 in HC + FENO group and HC group were not significantly different but were both significantly higher than that in control group (P < 0.01, P < 0.05). No changes were observed in Acox2, CYPSB1, and CYP27A1 mRNA levels among these three groups.

CONCLUSIONSimultaneous activation of LXR and PPARalpha can increase of CYP7A1 and DBP mRNA exDression and thus accelerates the biosynthesis of bile acid.

Animals ; Bile Acids and Salts ; biosynthesis ; Cholesterol ; pharmacology ; Fenofibrate ; pharmacology ; Hypolipidemic Agents ; pharmacology ; Liver ; drug effects ; enzymology ; Liver X Receptors ; Male ; Orphan Nuclear Receptors ; agonists ; PPAR alpha ; agonists ; Rats

[ABSTRACT]OBJECTIVEThe purpose of this study was to analyze the screened miRNAs related to the chemosensitivity for the TPF regimen of hypopharyngeal squamous cell carcinoma by miRNA array, and provide a set of miRNAs that may be useful for the development of novel diagnostic markers and more effective therapeutic strategies from the screened miRNAs.METHODSA total number of 21 patients who underwent TPF induction chemotherapy for primary hypopharyngeal squamous cell carcinoma were recruited for miRNA array analysis. 12 patients are sensitive to chemotherapy, and 9 patients are not. Moreover, the selected putative regulated miRNAs were also validated by RT-PCR in another 24 patients (14 patients are sensitive to chemotherapy, and others are not).RESULTSThere were 24 miRNA significantly differencial to the sensitivity to chemotherapy, and 6 miRNAs were up-regulated in the TPF group while 18 miRNA were down-regulated (P<0.05). To identify typical miRNA, mirfocus 3.0 database selected four miRNAs hsa-miR-211-3p, hsa-miR-4253, hsa-miR-4443, and hsa-miR-193b-3p, which were significant down-regulated in TPF-sensitive group. QRT-PCR further validated that only three miRNA (hsa-miR-4253、hsa-miR-4443、hsa-miR-193b-3p) were under-expressed in TPF-sensitive group of another 24 tissue samples (P<0.05).CONCLUSIONMiRNA hsa-miR-193b-3p, hsa-miR-4253, hsa-miR-4443 were identified in TPF-sensitive tissues by microarrays, and further validated by RT-PCR. These down-regulated miRNAs may act as novel biomarkers to classify TPF sensitivity of hypopharyngeal squamous cell carcinoma patients and will contribute to the understanding of the molecular basis of the chemosensitivity in the disease.

In this study, nattokinase gene was amplified by PCR using bacillus subtilis chromosomal DNA as template and cloned into expressed vector pBV220. After transforming recombinant plasmid into E.coli HB101, the recombinant strain was yielded. It was proved that expression products was secretive and expression protein was 12% of total cell protein by SDS-PAGE. Optimum culture time and inducing time was determined as 6h and 5h respectively. The plasmid stability studies showed that recombinant plasmid has excellent segregational stability but the structural stability was not good in the host cell.

OBJECTIVETo determine the physiological role of D-bifunctional protein (DBP) in bile acid biosynthesis through investigating the effect of increasing activity of DBP on bile acid biosynthesis.

METHODSTwenty male Wistar rats were divided into two groups: diethylhexyl phthalate (DEHP) group (n = 10) and control group (n = 10). Serum triglyceride, total cholesterol, hepatic DBP activity, and fecal bile acids were assayed. The mRNA levels of hepatic peroxisome proliferator-activated receptor alpha (PPARalpha), DBP, and cholesterol 7alpha-hydroxylase (CYP7A1) were detected by RT-PCR.

RESULTSCompared with control group, serum triglyceride level was decreased significantly and PPARalphamRNA level was increased significantly in DEHP group (P < 0.01). Together with a sharp induction of DBP mRNA expression and DBP activity in DEHP group (P < 0.01), the levels of CYP7A1 mRNA and fecal bile acids were significantly increased by 1.9 times and 1.6 times respectively compared to control group (P < 0.01). There was a significantly positive correlation between DBP mRNA level or DBP activity and CYP7A1 mRNA level (r = 0.89, P < 0.01; r = 0.95, P < 0.01).

CONCLUSIONThe up-regulation of DBP mRNA and activity in liver can result in the increase in CYP7A1 mRNA expression and bile acid biosynthesis, suggesting that DBP may be involved in bile acid biosynthesis together with CYP7A1.

17-Hydroxysteroid Dehydrogenases ; metabolism ; Animals ; Bile Acids and Salts ; biosynthesis ; Cholesterol 7-alpha-Hydroxylase ; analysis ; Enoyl-CoA Hydratase ; metabolism ; Liver ; metabolism ; Male ; Multienzyme Complexes ; metabolism ; PPAR alpha ; analysis ; Peroxisomal Multifunctional Protein-2 ; RNA, Messenger ; analysis ; Random Allocation ; Rats ; Rats, Wistar

OBJECTIVETo construct plant expression pCAMBIA1301-PMI by substituting PMI for hygromycin resistance gene in pCAMBIA1301 and obtain transgenic Salvia miltiorrhiza f. alba using PMI-mannose selection system.

METHODThe 6-phosphomannose isomerase gene (PMI) of Escherichia coli was amplified by PCR. Sequence analysis showed that it shared 100% amino acids identities with the sequences of PMI genes isolates reported in the NCBI. Based on pCAMBIA1305, the plant expression pCAMBIA1305-PMI was constructed successfully by substituting PMI for hygromycin resistance gene in pCAMBIA1305. pCAMBIA1305-PMI was transformed into Agrobacterium tumefaciens LBA4404, and then the leaves of S. miltiorrhiza f. alba were inoculated in LBA4404 with pCAMBIA1305-PMI.

RESULTPlant expression pCAMBIA1301-PMI was successfully constructed and the leaves of S. miltiorrhiza f. alba inoculated in LBA4404 with pCAMBIA1305-PMI were selected on medium supplemented with a combination of 20 g x L(-1) mannose and 10 g x L(-1) sucrose as a carbon source. The transformation efficiency rate was 23.7%.

CONCLUSIONGenetic transformation was confirmed by PCR, indicating that a new method for obtaining transgenic S. miltiorrhiza f. alba plants was developed using PMI-mannose selection system.

Anti-Bacterial Agents ; pharmacology ; Biomarkers ; Cinnamates ; pharmacology ; Escherichia coli ; enzymology ; genetics ; Escherichia coli Proteins ; genetics ; metabolism ; Gene Expression ; Genetic Vectors ; genetics ; metabolism ; Hygromycin B ; analogs & derivatives ; pharmacology ; Mannose-6-Phosphate Isomerase ; genetics ; metabolism ; Plants, Genetically Modified ; drug effects ; genetics ; metabolism ; Salvia miltiorrhiza ; drug effects ; genetics ; metabolism ; Transformation, Genetic

ATP-Binding Cassette, Sub-Family B, Member 1 ; metabolism ; Adolescent ; Adult ; Aged ; Aged, 80 and over ; Cell Nucleus ; metabolism ; Cytoplasm ; metabolism ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Lymphoma, Large B-Cell, Diffuse ; metabolism ; pathology ; Male ; Middle Aged ; Neoplasm Invasiveness ; Neoplasm Staging ; Prognosis ; Proliferating Cell Nuclear Antigen ; metabolism ; Y-Box-Binding Protein 1 ; metabolism ; Young Adult

OBJECTIVE The purpose of this study was to analyze the screened miRNAs related to tumorigenesis using miRNA array in laryngeal squamous cell carcinoma (LSCC), and provide a set of miRNAs that may be useful for the development of novel diagnostic markers and/or more effective therapeutic strategies from the screened miRNAs in LSCC. METHODS A total number of 5 patients who underwent surgery for primary laryngeal squamous cell carcinoma were recruited for miRNA array analysis. LSCC tissues compared with corresponding adjacent non-neoplastic tissues were analyzed by the Affymetrix GeneChip miRNA Array 3.0 to screen effective miRNAs, and the raw dataset had been submitted to Gene Expression Omnibus. Then mirfocus 3.0 database was adopted to analyze putative regulated miRNAs related to MCM4, a gene related to tumorigenesis we had studied previously in LSCC. Moreover, the selected putative regulated miRNAs were also validated by qRT-PCR in another 21 patients diagnosed as LSCC. RESULTS Analyzed by the miRNAs arrays, there were 127 miRNAs significantly related to tumorigenesis, and 78 showed a higher expression in tumor than in non-tumor tissue while 49 presented the contrasting pattern (P<0.01). Then analyzed by mirfocus 3.0 database, there were 2 putative regulated miRNAs, hsa-miR-24-3p and hsa-miR-183-5p, related to the expression of MCM4. Another miRNA we should focus on was hsa-miR-30a-5p, which was down-expressed obviously analyzed by the miRNA array. The expression of the 3 putative regulated miRNAs were also validated by qRT-PCR in another 21 patients, and the result was the same with that in miRNA array (P<0.05). CONCLUSION The 3 putative miRNAs based on miRNA array analysis, hsa-miR-24-3p, hsa-miR-183-5p and hsa-miR-30a-5p, could be considered as potential diagnostic and therapeutic markers in LSCC. The result will contribute to the understanding of the molecular basis of LSCC and help to improve the treatment.

Objective To investigate the effect ofeicosanoic acid (EA) on β-amyloid precursor protein (APP) and β-site APP cleaving enzyme 1 (BACE1) expressions in cortical neurons of primary cultured rats.Methods Cortical neurons of primary cultured rats were divided randomly into EA treatment group (being administered with 40 μmol/L eicosanoic acid for 24 h) and control group (medium containing the same amount of solvent).The APP and BACE1 protein levels were detected by Westem blotting.The APP and BACE1 mRNA levels were detected by reverse transcription-polymerase chain reaction (RT-PCR).Results The levels of APP751+770 mRNA,APP protein and BACE1 protein in the EA treatment group were significantly higher than those in control group (P＜0.05).Conclusion The saturated fatty acids increase the APP and BACE1 expressions,which may play an important role in the pathogenesis of Alzheimer's disease.

BACKGROUNDSuccessful periodontal regeneration depends on the migration, proliferation and differentiation of periodontal ligament cells in periodontal defects. The total protein content and the ultrastructure demonstrate the metabolizability and activity of periodontal ligament cells. This study was conducted to observe the effects of Shuanghuangbu, a mixture of medicinal herbs, on the total protein content and the ultrastructure of human periodontal ligament cells.

METHODSPeriodontal ligament cells were grown to confluence and then cultured in Dulbecco's modified eagle medium (DMEM) supplemented with Shuanghuangbu over the concentration range of 0 to 1000 microg/ml. The total protein content in cultured cells was determined by using Coommasie brilliant blue technique. Periodontal ligament cells were incubated in 0 and 100 microg/ml Shuanghuangbu decoction for 5 days, then observed through transmission electron microscope.

RESULTSThe total protein content of human periodontal ligament cells increased in each experiment group added 10 - 1000 microg/ml Shuanghuangbu respectively, and the effect of 100 microg/ml was excellent. Under the transmission electron microscope, there were more rough endoplasmic reticulums and mitochodrias in the experiment group than those in the control group.

CONCLUSIONShuanghuangbu stimulates the protein synthesis of human periodontal ligament cells and improves human periodontal ligament cells' metabolizability and activity.

Cells, Cultured ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Immunohistochemistry ; Microscopy, Electron, Transmission ; Periodontal Ligament ; chemistry ; drug effects ; ultrastructure ; Proteins ; analysis