AIMTo investigate the liver X receptors agonists T0901317's effect on expression of FAT/CD36 gene mRNA in adult human skeletal muscle cell.

METHODSMyotubes from humans were exposed to different T0901317 concentrations (0, 0.5, and 1.0 micromol/L) for 24 hours before experiments were performed. Then the expression of FAT/CD36 mRNA in skeletal muscle cell of each experimental group was detected by SYBR Green I real-time quantitative polymerase chain reaction. The relative data were compared among groups by 2-delta delta Ct method.

RESULTS(1) The Ct mean of control group, T0901317 (0.5 micromol/L) group, T0901317 (1 micromol/L) group were analyzed and there was significant difference (P < 0.01). (2) The expression of FAT/CD36 mRNA with liver X receptors agonists T0901317 in human skeletal muscle cell in the T0901317 (0.5 micromol/L) group and T0901317 (1 micromol/L) group were 2.91 times and 3.03 times than the control group.

CONCLUSIONThe expression of FAT/CD36 mRNA in human skeletal muscle cell afer the treatment of liver X receptors agonists T0901317 is increased, so we may propose that T0901317 may increase the risk of resistance in adult human skeletal muscle.

Adult ; CD36 Antigens ; genetics ; metabolism ; Cells, Cultured ; Female ; Humans ; Hydrocarbons, Fluorinated ; pharmacology ; Liver X Receptors ; Male ; Muscle, Skeletal ; cytology ; metabolism ; Orphan Nuclear Receptors ; agonists ; RNA, Messenger ; genetics ; metabolism ; Sulfonamides ; pharmacology

Objective To investigate the gene mutations of calcium-and integrin-binding protein 2 (CIB2) 196C＞T, 272T ＞ C and 297C ＞ G carried by students with non-syndromic hearing impairment from special educational schools in Yunnan Province. Methods The experimental group included 337 students with non-syndromic hearing impairment who failed to carry deafness gene with GJB2 (35 del G, 176_191 del 16,235delC, 299_300 del AT), GJB3 (C538T,G547A), mtDNA 12S rRNA (A1555G, C1494T), and SLC26A4 (IVS7_2A＞G, A2168G) . The control group consisted with 150 healthy people. Genomic DNA was isolated from peripheral blood with EDTA anti-coagulate. The subject's DNA fragments including CIB2 196C＞T, 272T ＞ C and 297C＞ G were amplified by polymerase chain reaction (PCR), and subsequently analyzed by direct sequencing to identify deafness-associated mutations. Results Both in the experimental group and control group, we failed to find the mutation of CIB2 196C＞T, 272T＞C and 297C＞G in all individuals. Conclusion Mutations in CIB2 gene 196C＞T, 272T＞C and 297C＞G are not a frequent cause of non-syndromic hearing loss among deaf people in Yunnan province. It provided important information for deafness with formulating landscape of gene screening in this region.

Exosomes are bilayer-lipid membrane nanovesicle from almost all living cell types which are in-volved in intercellular substance transporting and signaling communication .Exosomes are 30 ~120 nm in diameter , can transfer bioactive molecules including DNA , RNA, microRNA, protein as well as lipids derived from parents ' cells to re-cipient cells by body fluids , and specifically influence their physiological or pathological conditions .Leukemia is due to malignant proliferation of hematopoietic stem and progenitor cells .It was reported that leukemia cells derived exosomes play a key role in disease progression , drug resistance , and predict prognosis .This paper will outline the role of exosomes de-rived from leukemia cells and provide important information to help explore the molecular pathogenesis , biomarker as well as therapeutic target of leukemia .

Objective To explore the relationship between the volatile components in Angelicae Sinensis Radix from different regions of Gansu Province and its growing environment with metabolomics based on GC-MS. Methods The GC-MS method was used for detecting the volatile components in Angelicae Sinensis Radix from 31 different regions in Gansu province, and principal component analysis (PCA) and partial least squares (PLS) methods were used for analyzing and evaluating its relationship with the growing environment. Results The results of PCA showed that the volatile components in Angelicae Sinensis Radix from different regions in Gansu province were related to the altitude and the soil types. The PLS method could divide 31 samples of Angelicae Sinensis Radix from different regions in Gansu Province into three groups according to the difference of altitude. There were significant differences in the volatile components in the samples taken at different altitude regions. After analyzing linear loading plots from PCA and PLS, 11 charateristic components were screened out, including 7 compounds were identified by the retrieval of NIST11 database. Conclusion The volatile components in Angelicae Sinensis Radix from different regions in Gansu Province are closely related to the altitude and the soil type.

Objective To observe the clinical effects of Leigongtong-duogan Pian combine with total glucosids of Paeony Capsules to treat chronic idiopathic urticaria.Methods One hundred and thrity -eight patients are observed in this test , and they are divided into 3 groups.Group 1:were given Leigongtongduogan Pian 10 mg orally , 3 times a day and total glucosids of Paeony Capsules 600 mg orally, 3 times a day.Group 2:were given mizolastine 10 mg orally, onces a day;and total glucosids of Paeony Capsules 600 mg orally, 3 times a day.Group 3:were given mizolastine 10 mg orally onces a day.Duration in the three groups were for 28 days, if someone recovers within 28 days, he also continues until the end.The itching intensity and amount of rush and the size of rush , even the frequency of the outbreaks of rush are observed on the 7th day and on the 14th day and on the 28th day.Results The clin-ical effective rate of group 1 is 91.30%, however group 2 is 77.14%and group 3 is 71.43%.Conclusion The clinical effects of Leigong-tongduogan Pian combine with total glucosids of Paeony Capsules to treat chronic idiopathic urticaria are remaked and its adverse drug reactions are lowered.

Primer Express 2.0 software was used to design the primers and the MGB probe. With the plasmid pHaMDR1/A containing mdr1 cDNA as the template, we established a real-time fluorescent quantitative polymerase chain reaction system, which, at the template concentration of 3.061 x 10(3) to 3.061 x 10(9) cps/ml, had a correlation coefficient of 0.988243 between template concentration and threshold cycle value. This PCR method allows sensitive, specific and quantitative detection of human mdr1 gene.
ATP-Binding Cassette, Sub-Family B, Member 1 ; analysis ; genetics ; DNA Primers ; Female ; Fluorescent Dyes ; Fluorometry ; methods ; Genes, MDR ; genetics ; Humans ; Male ; Polymerase Chain Reaction ; methods ; Taq Polymerase

Endometriosis(EMs), whose pathogenesis is complicatedand is not fully understood, is a common gynecological disease. The association between gene polymorphism and EMs is a hot spot of research for its pathogenesis and pathogenic mechanism, which provides a research basis for detection of susceptible disease loci inhigh-risk groups and the identification and genetic analysis ofdiseases and related genes, and offers more help for EMs patients in clarifying diagnosis at source and improving therapy outcome. This paper reviews the research status of EMs gene polymorphism.

Objective To investigate the effect of knockdown of EpCAM by siRNA on invasion,migration,and colony abilities in hypopharyngeal carcinoma FaDu cells.Methods A siRNA against EpCAM was employed to inhibit the expression of EpCAM in FaDu cells.Measurements included the Transwell assay for invasion and migration,plate colony formation assay for cell colony ability,Western blot assay for EpCAM,E-cadherin,and β-catenin expressions in total protein,cytoplasm,and cytoskeleton,respectively.Results mRNA and protein expressions of EpCAM were suppressed significantly in FaDu cells transfected by EpCAM siRNA (t =6.46,P ＜ 0.05 ; t =10.25,P ＜ 0.05).Transwell assay showed in transwell assay,the average invasive cells in EpCAM siRNA cells (26.33 ± 3.71) was less than that in FaDu cells (61.47 ± 6.70 ; t =7.95,P ＜ 0.05) and control cells (54.13 ± 6.51 ; t =6.42,P ＜ 0.05) ; the average number of migration cells in EpCAM siRNA cells (79.87 ± 8.44) was lower than that in FaDu (167.53 ± 11.49 ; t =10.90,P ＜ 0.05) cells and control cells (162.13 ± 13.45 ; t =8.97,P ＜ 0.05).In plate colony formation assay,the average colony number of EpCAM siRNA cells was (78.00 ± 5.57),which was less than that of FaDu cells(177.30 ± 16.50; t =9.78,P ＜0.05) and control cells (173.67 ± 13.50; t =11.35,P ＜0.05).Western blot assays showed,silencing of EpCAM increased the expressions of E-cadherin (t =4.58,P =0.01) and β-catenin (t =3.76,P =0.02) in cytoskeleton,and decreased the expressions of E-cadherin (t =6.60,P ＜ 0.05) and β-catenin (t =8.20,P ＜ 0.05) in cytoplasm.Conclusions The knockdown of EpCAM inhibits the invasion,migration,and colony formation abilities of FaDu cells,which is probably related to the regulation of E-cadherin and β-catenin in cytoplasm and cytoskeleton,and EpCAM may be a promising gene therapy target for hypopharyngeal carcinoma.

OBJECTIVETo investigate the relationship between calcium-sensing receptor protein (CaSR) expression and rat cardiomyocyte apoptosis and related signal transduction pathways.

METHODSThe CaSR, BCl2, Caspase3 protein and ERK1/2 phosphorylation or non-phosphorylation were detected by Western blot. Cardiomyocyte apoptosis was detected by flow cytometry and immunofluorescence.

RESULTSCaSR protein was detected in rat cardiac tissue and CaSR activator gadolinium (GdCl3) induced cardiomyocyte apoptosis and increased ERK1/2 phosphorylation and expression of BCl2 and activated Caspase3. The selective mitogen-activated protein kinase (MAPK) inhibitor PD98059 abolished gadolinium -induced ERK1/2 activation and BCl2 expression, further increased the activation of Caspase3 and cardiomyocyte apoptosis.

CONCLUSIONOur results demonstrate the CaSR existence in cardiomyocytes and CaSR activation by gadolinium can induce myocyte apoptosis by activating Caspase3 and tyrosine protein kinase pathway.

Animals ; Apoptosis ; Caspase 3 ; metabolism ; Female ; Male ; Myocardium ; metabolism ; Myocytes, Cardiac ; metabolism ; RNA, Messenger ; biosynthesis ; Rats ; Rats, Wistar ; Receptors, Calcium-Sensing ; biosynthesis ; genetics ; Signal Transduction

ObjectiveExosomes secreted by BMSC overexpressing GATA-4 gene (BMSCGATA-4-exosome) can promote the differentiation of BMSC into cardiomyocyte-like cells, thereby improve cardiac function after myocardial infarction. However, the molecular mechanism of BMSCGATA-4-exosome in cardiomyocyte-like cell differentiation is unknown. The effect of the secretion of BMSCGATA-4 exosome from bone marrow mesenchymal stem cells (BMSC) in the differentiation of stem cells into cardiomyocytes was determined in miRNA-673-5p/Tsc-1 axis dependent manner.MethodsMouse models of myocardial infarction were established and divided into seven groups. Simulation group (BMSCmiR-673-5p-mimic exosome), inhibition group (BMSCmiR-673-5p-inhibitor exosome), GATA-4 group (BMSCGATA-4 exosome), empty vector group (BMSCempty vector exosome）, and BMSC group (BMSC exosome) were injected into the tail vein for 48 h, and the untreated and normal mice were used as the control group. Cardiac ultrasound was used to detect cardiac function in each group. miRNA-673-5p expression in myocardial infarction was detected using real-time polymerase chain reaction (RT-PCR). The myocardial tissues were extracted from the same myocardial infarction site. Myocardial-specific molecules, such as α-actin, Desmin, cTnT, and Cx43, were detected using RT-PCR. Western blot was used to determine the expression of the corresponding target gene of miRNA-673-5p, Tsc-1, Erk1/2, and Mef2c proteins.ResultsThe simulation group wan shown the most significantly improved myocardial function (P<0.05) with an expression peak of miRNA-673-5p in cardiomyocytes (P<0.05). The highest content of myocardial-specific molecules including α-actin, Desmin, cTnT, and Cx43 was found in the simulation group. The simulation group had the lowest expression of Tsc-1 in cardiomyocytes (P<0.05).ConclusionOverexpressed BMSCGATA-4 exosomes inhibit Tsc-1 expression through miRNA-673-5p to improve cardiac function during myocardial infarction.