OBJECTIVETo study the expression of the receptor for advanced glycation end products (RAGE) and the inhibitory effect of advanced glycation end products (AGEs) on testosterone production in rat Leydig cells.

METHODSRat Leydig cells were primarily cultured and the expression of RAGE in the Leydig cells was detected by RT-PCR and immunofluorescence staining. The Leydig cells were treated with AGEs at the concentrations of 25, 50, 100 and 200 microg/ml, respectively, and the testosterone content was determined by ELISA.

RESULTSRT-PCR and immunofluorescence staining exhibited the expression of RAGE in the rat Leydig cells. AGEs remarkably suppressed hCG-induced testosterone production in the Leydig cells in a concentration-dependent manner in the 50, 100 and 200 microg/ml groups as compared with the control (P < 0.01).

CONCLUSIONRAGE exists in rat Leydig cells and AGEs can significantly inhibit the secretion of testosterone in primarily cultured rat Leydig cells.

Animals ; Cells, Cultured ; Enzyme-Linked Immunosorbent Assay ; Glycation End Products, Advanced ; pharmacology ; Leydig Cells ; metabolism ; radiation effects ; Male ; Rats ; Receptor for Advanced Glycation End Products ; Receptors, Immunologic ; biosynthesis ; Reverse Transcriptase Polymerase Chain Reaction ; Testosterone ; biosynthesis

Objective To investigate the effect of antisense oligonucleotides (ASODN) targeting livin on the inhibition of livin mRNA and protein expression and the apoptosis of human renal carcinoma cell line 786-O cells. Methods Specific phosphorothioate antisense oligodeoxynucleotides targeting livin were synthesized and then transfected into 786-O cells. The expressions of livin mRNA were detected by RT-PCR. Expression and location of livin protein were observed by confocal laser scanning microscope (CLSM). Apoptosis rate of 786-O cells was investigated by flow cytometer. The activity of Caspase-3 was detected by colorimetric assay. Results After the transfection of ASODN, the expression of livin mRNA was decreased (P

OBJECTIVETo introduce the location and course of S1, S2 sacral nerve root tunnel and to clarify the significance of the anterior aspect of sacral nerve root tunnel on placement of iliosacral screw on the standard lateral sacral view.

METHODSFirstly the data of 2.0 mm slice pelvic axial CT images were imported into Mimics 10.0, and the sacrum, innominate bones, and sacral nerve root tunnels were reconstructed into 3D views respectively, which were rotated to the standard lateral sacral views, pelvic outlet and inlet views. Then the location and course of the S1, S2 sacral nerve root tunnel on each view were observed.

RESULTSThe sacral nerve root tunnel started from the cranial end and anterior aspect of the vertebral canal of the same segment and ended up to the anterior sacral foramen with a direction from cranial-posterior-medial to caudal-anterior-lateral. The tunnel had a lower density than the iliac cortex and greater sciatic notch on the pelvic X-rays,especially on the standard sacral lateral view, on which it showed up as a disrupted are line and required more careful recognition.

CONCLUSIONIt can prevent the iliosacral screw from penetrating the sacral nerve root tunnel and vertebral canal when recognizing the anterior aspect of sacral nerve root tunnel and choosing it as the caudal-posterior boundary of the "safe zone" on the standard lateral sacral view.

Adult ; Aged ; Bone Screws ; Female ; Fracture Fixation, Internal ; Fractures, Bone ; surgery ; Humans ; Male ; Middle Aged ; Pelvic Bones ; diagnostic imaging ; injuries ; innervation ; surgery ; Radiography ; Sacrococcygeal Region ; diagnostic imaging ; innervation ; surgery ; Sacrum ; diagnostic imaging ; injuries ; innervation ; surgery ; Spinal Nerve Roots ; diagnostic imaging ; surgery ; Young Adult

Adult ; Aged ; Bone Plates ; Female ; Femoral Fractures ; surgery ; Femur ; injuries ; surgery ; Fracture Fixation, Internal ; Humans ; Male ; Middle Aged ; Minimally Invasive Surgical Procedures

Cell Differentiation ; Gene Expression Regulation ; Liver ; cytology ; Stem Cells ; cytology

Little is known about the non-structural protein 6(NSP6)of rotavirus.This report describes expression of the NSP6 of a group A human rotavirus strain TB-Chen in bacteria,and its immunological properties and cellular distribution.The results showed that the recombinant NSP6(rNSP6)was expressed in high efficiency without any other proteins fused(possesses about 34.2% of total bacterial proteins).rNSP6 elicited mono-specific antibodies in immunized guinea pigs and the antibodies could react with the rNSP6 itself and the viral NSP6 proteins synthesized in SA11-or Wa-infected MA104 cells in Western blot and immunofluorescence assay.The NSP6 distributed evenly in the cytoplasm mainly around the nucleus of virus-infected cells,no viroplasm-like gathering observed;The top amount of NSP6 synthesized in SA11-infected cells or Wa-infected cells could be detected at 12h after infection.This is the first report about the high expression of entire NSP6(without any other proteins fused)in prokaryotic expression system and detection of NSP6 synthesis in virus infected cells by immunofluorescence assay.The results are important to understand the structure,biological properties and further application of the NSP6.

Objective To construct replication-deficient recombinant adenoviruses AdHNF4?that co-expresse human hepatocyte nuclear factor 4?(HNF4?) and green fluorescent protein(GFP) gene,and to evaluate the effect of HNF4?up-regulation on hepatocyte gene expression.Methods The HNF4?cDNA was obtained through RT-PCR from human hepatocyte.The recombinant adenoviral plasmid- pAdHNF4?was established using AdEasy system and packed in 293 cells.After transfection of human hepatoma cell lines HepG2 and Hep3B with AdHNF4?,the expression of HNF4?and other liver-associ- ated functional genes were evaluated by RT-PCR and Western blot.Results The recombinant plasmid pAdHNF4?was confirmed by restriction endonuclease digestion and sequencing.GFP expression was observed on the fourth day after packing the linearized pAdHNF4?in 293 cells.Stable transfection of AdHNF4?with a titer of 1?10~(10) efu/ml was obtained after repeated amplification.More than 90% of human hepatoma cells had GFP expression in 72 hours after transfection of AdHNF4?.The expression of HNF4?mRNA and protein were significantly up-regulated compared with the control group(3.4 folds in HepG2 infected with AdHNF4a and 5.2 folds in Hep3B infected with AdHNF4?).Furthermore,the transcriptional expressions of some liver-associated functional genes such as apolipoprotein,cytochrome P450 families,glutamine synthetase,phosphoenolpyruvate carboxykinase and glucose-6-phosphatase also increased after transfection of the virus,and the apoptosis ratio of the cells increased.Conclusions Up- regulating the expression of HNF4?in human hepatoma cells with AdHNF4?could enhance normal liver- specific function.Our study would provide a new idea for the researches on gene regulation of transplan- ted hepatocytes.

Objective：Rat UREB1 protein coded by the gene UREB1 can specially bind to URE (upstream regulatory element) which is in the upstream of the promoter. It′ s reported that the protein of UREB1 promote the transcription of Dynorphin gene and inhibits p53 transactivation. This study was designed to clone human UREB1 gene and explore the relationship between UREB1 and the development of tumor. Methods： The artificial synthetic oligonucleotide was used as the probe to screen human brain cDNA library and human UREB1 gene was cloned. The antibody, which was produced using the recombinant UREB1 from E.coli as the antigen and immunizing the animals, was utilized for detecting the distribution of UREB1 in different tumor tissues. Results： The human UREB1 gene was cloned by using in situ hybridization for screening human brain cDNA library, and the nucleotide sequences and the deduced amino acid sequence of human UREB1 has 91％ homology with that of rat UREB1 identified previously. Western blot analysis revealed that the human UREB1 was present in all tumor tissues but the quantity of UREB1 in different tissues was not the same. Immunohistochemistry results shown that the human UREB1 distributes primarily in the cytoplasm and nuclear of tumor cells and nuclear UREB1 in carcinosarcoma is much more than that in adenoma. After analyzing the level of tyrosine phosphorylated UREB1 in a few tumor tissues, the result shown that the more malignant the tumor tissue was, the higher level the tyrosine phosphorylated of UREB1 was in that tumor tissues. Conclusion： Human UREB1 may be involved in the development of tumor and its tyrosine phosphorylation may affect the degree of tumor malignant.

BACKGROUNDDermatomyositis (DM) and polymyositis (PM) are common inflammatory myopathies whose immunopathogenic mechanisms remain poorly understood. The NOD-like receptor family, pyrin domain containing 3 (NLRP3) inflammasome is a type of cytoplasmic multiprotein inflammasome and is responsible for the activation of inflammatory reactivations. Responding to a wide range of exogenous and endogenous microbial or sterile stimuli, NLRP3 inflammasomes can cleave pro-caspase-1 into active caspase-1, which processes the pro-inflammatory cytokines pro-interleukin (IL)-1β and pro-IL-18 into active and secreted IL-1β and IL-18. The NLRP3 inflammasome is implicated in infectious and sterile inflammatory diseases. However, it remains unclear whether it is involved in the pathogenesis of DM/PM, which we aim to address in our research.

METHODSIn this study, 22 DM/PM patients and 24 controls were recruited. The protein and RNA expression of IL-1β, IL-18, NLRP3, and caspase-1 in serum and muscle samples were tested and compared between the two groups.

RESULTSThe serum IL-1β and IL-18 levels were significantly higher in DM/PM patients than those in the controls by enzyme linked immunosorbent assay (ELISA, DM vs. control, 25.02 ± 8.29 ng/ml vs. 16.49 ± 3.30 ng/ml,P < 0.001; PM vs. control, 26.49 ± 7.79 ng/ml vs. 16.49 ± 3.30 ng/ml,P < 0.001). Moreover, the real-time quantitative reverse transcription-polymerase chain reaction (qRT-PCR) showed that DM/PM patients exhibited higher RNA expression of IL-1β, IL-18, and NLRP3 in the muscle (for IL-1β, DM vs. control, P= 0.0012, PM vs. control, P= 0.0021; for IL-18, DM vs. control, P= 0.0045, PM vs. control, P= 0.0031; for NLRP3, DM vs. control, P= 0.0017, PM vs. control, P= 0.0006). Moreover, the protein expression of NLRP3 and caspase-1 in muscle samples of DM/PM patients were also significantly elevated compared to that in the muscles of the controls.

CONCLUSIONSOur findings demonstrate that the NLRP3 inflammasome is implicated in the pathogenesis of DM/PM. High NLRP3 expression led to elevated levels of IL-1β and IL-18 and could be one of the factors promoting disease progress.

Adult ; Caspase 1 ; analysis ; genetics ; Dermatomyositis ; etiology ; Female ; Humans ; Inflammasomes ; physiology ; Interleukin-18 ; analysis ; genetics ; Interleukin-1beta ; analysis ; genetics ; Male ; Middle Aged ; NLR Family, Pyrin Domain-Containing 3 Protein ; analysis ; genetics ; physiology ; Polymyositis ; etiology

OBJECTIVETo compare the effect of Acanthopanacis senticosi injection, theophylline and caffeine on human sperm mobility in vitro.

METHODSWe incubated the sperm aseptically obtained by masturbation from 12 asthenospermia men and treated by swim-up technique in Acanthopanacis senticosi injection (10 g/L), theophylline (3 mmol/L) and caffeine (7 mmol/L) respectively, and detected various sperm parameters with the computer-assisted sperm analysis (CASA) system at 0 h, 1 h and 3 h.

RESULTSAcanthopanacis senticosi injection significantly increased sperm motility, the percentage of progressive motile sperm, straight line velocity (VSL) and curvilinear velocity (VCL) as compared with theophylline and caffeine (P < 0.05).

CONCLUSIONAcanthopanacis senticosi injection can activate the mobility of human sperm in vitro.

Caffeine ; pharmacology ; Drugs, Chinese Herbal ; pharmacology ; Eleutherococcus ; chemistry ; Humans ; In Vitro Techniques ; Male ; Sperm Motility ; drug effects ; Theophylline ; pharmacology