Diagnosis, Differential ; Humans ; Hyponatremia ; diagnosis ; etiology ; therapy ; Inappropriate ADH Syndrome ; diagnosis ; etiology ; therapy

OBJECTIVETo investigate the effects of Salvianolic acid B preconditioned endothelial progenitor cells (EPCs) on the Nkx2.5 and GATA-4 gene expressions at the early stage of cell differentiation of bone mesenchymal stem cells (BMSc) transplanted into infarcted myocardium, in order to find out the best synergism for co-transplantation of the two kinds of cells.

METHODSBMSc and EPCs of rats were isolated and cultured, and rats were modeled into acute myocardial infarction (AMI) by left coronary artery ligation. Then the EPCs preconditioned with different concentrations of Salvianolic acid B and BMSc or DMEM medium were implanted into heart ischemia area. Expressions of Nkx2.5 and GATA-4 mRNA expressions in myocardium were detected by Real-time RT-PCR 4 weeks later.

RESULTSCompared with those in the non-implanted model rats' myocardium, the gene expression of Nkx2.5 and GATA-4 mRNA were significantly higher in all the transplantation receptive groups, comparisons between the implanted groups showed that the highest value of expressions (2. 654 +/- 0.606 of Nkx2.5 and 1.573 +/- 0.372 of GATA-4) displayed in the group contained more EPCs, for 8-fold to BMSc in volume.

CONCLUSIONBMSc can differentiate into cardiac muscle like cells, and condition of their differentiation is related with the degree of the internal environment improved.

Animals ; Benzofurans ; pharmacology ; Cells, Cultured ; Endothelial Cells ; cytology ; drug effects ; transplantation ; Gene Expression ; drug effects ; Male ; Mesenchymal Stem Cell Transplantation ; Myocardial Infarction ; metabolism ; therapy ; Myocardium ; metabolism ; Random Allocation ; Rats ; Rats, Wistar ; Stem Cell Transplantation ; Stem Cells ; cytology ; drug effects

OBJECTIVETo investigate the characteristics of pain in temporomandibular disorders (TMD) through analyzing the description of pain by the TMD patients.

METHODSNinety TMD pain patients were included and the glossary in description of the intensity, rhythm and degree of disability due to TMD pain were recorded.

RESULTSThe descriptive pain by 90 patients was slight to moderate. There was no significant difference between males and females or between chronic and acute patients in the description of pain intensity (P > 0.05). The chewing disability was the most often reported complaining, and then was mental status. The probability of pain at rest was not different between in chronic pain patients and acute pain patients(P > 0.05). The acute pain patients often used aching, slight and tingle to describe their pain, while the chronic pain patients used dull, gas and numb more.

CONCLUSIONPain intensity in acute or chronic TMD patients is both low. The pain mainly influences chewing function and mental status in patients. Descriptive characteristic with chronic orofacial pain is different from acute.

Adult ; Facial Pain ; Female ; Humans ; Male ; Mastication ; Pain ; Temporomandibular Joint Disorders

OBJECTIVETo investigate the effects of X-ray exposure on the release of soluble tumor necrosis factor receptor-p75 (sTNFR-p75) in hepatocellular carcinoma HepG2 cells in vitro.

METHODSEnzyme-linked immunosorbent assay (ELISA) was used to examine the levels of sTNFR-p75 in the supernatants of HepG2 cells before and after X-ray exposure. The cell apoptosis was analyzed by flow cytometry and transmission electron microscope(TEM), and the morphological changes of the cells were examined under optical microscope and transmission electron microscope(TEM).

RESULTSX-ray exposure of the cells resulted in a strong increase of cell apoptosis (P<0.05) and sTNFR-p75 production in the cells as compared with the those before the exposure (P<0.01). Optical microscopy revealed apoptotic changes of HepG2 cell after the exposure, shown as cell shrinkage, spherical cell morphology, cytoplasmic and nuclear condensation. Apoptotic bodies were detected by TEM.

CONCLUSIONX-ray exposure induces HepG2 cells apoptosis by inhibiting the release of sTNFR-p75 into the supernatant.

Animals ; Apoptosis ; radiation effects ; Carcinoma, Hepatocellular ; pathology ; secretion ; Cell Line, Tumor ; Culture Media, Conditioned ; chemistry ; metabolism ; radiation effects ; Humans ; Liver Neoplasms ; pathology ; secretion ; Microscopy ; Receptors, Tumor Necrosis Factor, Type II ; biosynthesis ; chemistry ; secretion ; Solubility ; X-Rays

OBJECTIVETo construct a vector expressing small interfering RNA (siRNA) against Rac1 gene and observe its effect on soft agar colony formation of SW480 cells in vitro.

METHODSOligos of 64 base pairs for hairpin RNA targeting Rac1 were chemically synthesized and annealed. The siRNA constructs for Rac1, produced by inserting the annealed oligos into the downstream of H1 promoter of linearized pSUPER, were confirmed by restriction digestion and DNA sequencing. The constructed Rac1-siRNA was transfected into SW480 cells and Western blotting was performed to assess the expression and interference efficiency of siRNAs against Rac1.The soft agar colony formation assay was used to study the effect of Rac1 gene silencing on SW480 cells.

RESULTSRestriction digestion and DNA sequencing showed that the siRNA targeting Rac1 gene was successfully constructed. The siRNA could effectively down-regulate the expression of Rac1 in SW480 cells. Soft agar colony formation assay showed that the colony number and diameter of SW480 cells was reduced after siRNA transfection.

CONCLUSIONA vector expressing hairpin RNA against Rac1 gene are successfully produced, which significantly reduces the colony numbers and size of SW480 cells in vitro, suggesting that Rac1 plays an important role in the growth of colorectal cancer in vitro.

Base Sequence ; Cell Line, Tumor ; Cell Proliferation ; Colonic Neoplasms ; pathology ; Down-Regulation ; Humans ; Molecular Sequence Data ; RNA Interference ; RNA, Small Interfering ; genetics ; Transfection ; rac1 GTP-Binding Protein ; genetics

BACKGROUNDMonilethrix is an autosomal dominant hair disorder characterized clinically by alopecia and follicular papules. In this study, we collected a Han monilethrix family to detect the mutations in patients and investigated the correlation between the genotype and phenotype of monilethrix.

METHODSIn this study, we identified a Chinese family with monilethrix through light microscopic and scanning electron microscopic (SEM) examination. Genomic DNA from peripheral blood samples was prepared. DNA samples from controls and monilethrix patients were subject to polymerase chain reaction (PCR) amplification. Two pairs of primers were used to amplify the seventh exon of KRT86. Mutation screening of the PCR products was detected using direct sequencing.

RESULTSLight microscopic examination showed a regular alternate enlargement and narrow area. SEM examination showed that part of the cuticle of the nodules shed and disappeared gradually in the narrow area with granular protrusions on the surface similar to the erosion-like structure. Parallel longitudinal ridge and groovepattern appeared, and the ridges varied in width, like dead wood. A heterozygous transversion mutation c.1204G > A (p.E402K) in the seventh exon of KRT86 was identified in both patients.

CONCLUSIONSThe mutation of extron 7 of KRT86 identified plays a major role in the pathogenesis of this pedigree with monilethrix, and is a mutation hot spot of KRT86. Further research is needed to explore the relationship between the phenotype and the mutation of the type II hair keratin gene KRT86 of monilethrix.

Asian Continental Ancestry Group ; genetics ; China ; ethnology ; Humans ; Keratins, Hair-Specific ; genetics ; Keratins, Type II ; genetics ; Microscopy, Electrochemical, Scanning ; Monilethrix ; etiology ; genetics ; pathology ; Mutation

OBJECTIVETo study the role of activation of Rac1 in colorectal cancer cell migration and invasion.

METHODSRac1 L61 plasmid and control plasmid were transfected into colorectal cancer cell line SW480 cells. Pull down assay by Western blotting was carried out to measure the amount of activited Rac1, and transwell permeable supports were used to assess the migration and invasion of SW480 cells with different activitivity of Rac1.

RESULTSThe transfection ratio of SW480 cells was more than 80%. Pull down assay showed that the activited Rac1 was significantly higher in the SW480 cells transfected with Rac1 L61 plasmid than that in the control, and the amount of migrating and invasing SW480 cells transfected with Rac1 L61 plasmid in the Transwell permeable supports were significantly more than those in controls (migrating cell numbers: 43 +/- 9 vs. 22 +/- 5, P < 0.01; invasing cell numbers: 73 +/- 13 vs. 38 +/- 1, P < 0.01).

CONCLUSIONThe activation of Rac1 plays an important role in the migration and invasion of colorectal cancer cells.

Cell Line, Tumor ; Cell Movement ; Colonic Neoplasms ; metabolism ; pathology ; Enzyme Activation ; Genetic Vectors ; Green Fluorescent Proteins ; genetics ; metabolism ; Humans ; Neoplasm Invasiveness ; Plasmids ; Recombinant Fusion Proteins ; genetics ; metabolism ; Transfection ; rac1 GTP-Binding Protein ; genetics ; metabolism

OBJECTIVETo clone migfilin-N terminal sequence into E.coli and obtain a fusion protein for preparing rabbit polyclonal antibody against migfilin, thereby facilitating the study of the role of migfilin in the biological behavior of colon cancer.

METHODSBased on human migfilin cDNA sequence, a pair of primers was designed to amplify migfilin-N terminal sequence by PCR. The PCR product was subcloned into the bacterial expression vector pGEX-4T-1 with EcoRI/XhoI sites, and the target recombinant plasmids were identified with enzymatic cleavage followed by DNA sequence analysis. By transforming the expression vector into component E.coli BL(21) cells, the GST-migfilin-N fusion protein was expressed with IPTG induction. Glutathione-sepharose beads were used to purify the fusion protein, and anti-migfilin polyclonal antibody was produced by immunization of rabbits with the purified GST-migfilin N-terminal fusion protein. The resultant anti-migfilin polyclonal antibody was purified by protein A beads and used for Western blotting for detecting migfilin expression in different cell lines.

RESULTS AND CONCLUSIONThe migfilin-N terminal gene fragment was cloned successfully, and purified GST-migfilin N-terminal fusion protein and anti-rabbit migfilin polyclonal antibodies were obtained. Western blot analysis demonstrates that the antibodies specifically detected migfilin expression in the cell lines, which may facilitate further investigation of the role of migfilin in the biology of colon cancer.

Animals ; Antibodies, Monoclonal ; biosynthesis ; immunology ; isolation & purification ; Base Sequence ; Blotting, Western ; Cell Adhesion Molecules ; genetics ; immunology ; Cell Line, Tumor ; Cloning, Molecular ; Colonic Neoplasms ; genetics ; metabolism ; pathology ; Cytoskeletal Proteins ; genetics ; immunology ; DNA, Complementary ; chemistry ; genetics ; Escherichia coli ; genetics ; Humans ; Molecular Sequence Data ; Rabbits ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, DNA

Single-Strand Conformation Polymorphism(SSCP) is an effect method for investigating environment microbial genetic polymorphism, with its characterization of rapidness, simplicity, and sensitivity. However, many factors can influence the results of SSCP in the analysis of complex environment samples, and its optimization is highly needed. In this paper, optimal PCR-SSCP conditions were discussed based on PAGE concentration, formamide deionized in denaturing loading buffer, electrophoresis time and temperature. The resluts showed that the optimal conditions were as follows: 16S rDNA V1~V3 was selected as the targeted gene, the ratio of acrylamide to N, N-dimethylacrylamide in 12% polyacrylamide gel electrophoresis(PAGE)gel was 49:1, the ratio of formamide deionized in denaturing loading buffer was 1:3, running the SSCP gel at 300 V for 18 h (under 4 ℃). Aside from this, the validations using samples from a simultaneous desulfurification and denitrification bioreactor were conducted under this optimal conditions.

Objective To investigate the effect of 1 800 MHz electromagnetic radiation on activity of SOD and GSH－Px in the skin tissues of SD rats.Methods A total of 98 healthy SD rats with SPF level,aged 4 weeks, were randomly divided into radiation group and control group.The radiation group was totally exposed under 1 800 MHz electromagnetic wave with seven different power density of radiation of 0.1 mW/cm2,0.3 mW/cm2,0.5 mW/cm2 , 0.7 mW/cm2, 0.9 mW/cm2, 1.0 mW/cm2and 1.2 mW/cm2respectively.It lasted 21 days and for a period of 12 hours a day. After radiation,the activity of SOD and GSH－Px in the skin tissues were detected by enzyme marker. Results In radiation group,the activity of SOD and GSH－Px in the skin tissues of SD rats were decreased under 0.3 mW/cm2and 0.5 mW/cm21 800 MHz electromagnetic wave. Compared with the control group, there was a significantly difference in radiation group (P＜0.05) .While under other four 1 800 MHz electromagnetic waves, the activity of GSH－Px and SOD in the skin tissues showed no statistical difference between the two groups (P＞0.05) . Under 1 mW/cm21 800 MHz electromagnetic wave, the activity of GSH－Px showed no statistical difference between two groups (P＞0.05) . Conclusion The power density of 0.3 mW/cm2and 0.5 mW/cm21 800 MHz electromagnetic wave can reduce the activity of GSH－Px and SOD in the skin tissues of rats.