The aim of this research is to evaluate the effect of tetramethylpyrazine (TMP) and connective tissue growth factor (CTGF) miRNA plasmids on the expressive levels of CTGF, transforming growth factor-beta (TGFbeta) and type I collagen of rat hepatic stellate cells (HSC) which are stimulated by high glucose. The rat HSCs which were successfully transfected rat CTGF miRNA plasmids and the rat HSCs which were successfully transfected negative plasmids were cultured in vitro. After stimulus of the TMP and the high glucose, the protein levels and gene expressive levels of CTGF, TGF-beta and type I collagen were tested. The results indicated that high glucose increased the expression of CTGF mRNA, CTGF protein, TGF-beta mRNA,TGF-beta protein and type I collagen (P < 0.05). The expressive levels of CTGF mRNA, CTGF protein, TGF-beta mRNA, TGF-beta and type I collagen in TMP group were lower than those in high glucose group and showed statistically significant differences (P < 0.05). Compared with high glucose group, the expressive levels of CTGF mRNA, CTGF protein, TGF-beta mRNA, TGF-beta and type I collagen in rat CTGF miRNA plasmid interference group were significantly lower (P < 0.05). However, no statistically significant difference was found in CTGF mRNA and CTGF protein levels between TMP group and CTGF miRNA group (P > 0.05), while type I collagen levels showed statistically significant differences (P < 0.05). It is concluded that high glucose could promote the expressions of CTGF, TGF-beta and type I collagen, and TMP and rat CTGF miRNA plasmids could reduce the expressions of CTGF, TGF-beta, type I collagen.
Animals ; Cells, Cultured ; Collagen Type I ; metabolism ; Connective Tissue Growth Factor ; genetics ; Culture Media ; pharmacology ; Glucose ; pharmacology ; Hepatic Stellate Cells ; drug effects ; metabolism ; MicroRNAs ; genetics ; Plasmids ; Pyrazines ; pharmacology ; RNA, Messenger ; Rats ; Transfection ; Transforming Growth Factor beta ; metabolism

Adenocarcinoma ; chemically induced ; pathology ; Animals ; Animals, Newborn ; Lung Neoplasms ; chemically induced ; pathology ; Mice ; Mice, Inbred BALB C ; Random Allocation ; Sterigmatocystin ; toxicity

OBJECTIVETo provide clinicians with data supporting three different clearance techniques in the obtunded patients after severe polytrauma.

METHODSThis study gave an overview of the available and pertinent literature regarding cervical spine clearance in obtunded patients after severe polytrauma.

RESULTSCurrently, there were three accepted techniques for clearance of the cervical spine in obtunded patients after severe polytrauma. Each of these methods has advantages and disadvantages to both of the patients and the clinicians.

CONCLUSIONSThere are continuous improvements in both computed tomography (CT) and magnetic resonance imaging (MRI) techniques that increase their sensitivities. The continued use of plain radiographs is called into question with respect to cost and time requirements. An algorithmic approach to the evaluation of the cervical spine in the obtunded patients will lead to fewer missed injuries.

Cervical Vertebrae ; injuries ; Humans ; Magnetic Resonance Imaging ; Multiple Trauma ; diagnosis ; Physical Examination ; Spinal Injuries ; diagnosis ; Tomography, X-Ray Computed ; Unconsciousness

Omega-3 polyunsaturated fatty acids (PUFAs) have been broadly investigated and shown to exert many preventive and therapeutic actions besides their important role in maintenances human health and normal development. In mammals, the level of omega-3 PUFAs is relatively too low compared with omega-6 PUFAs, which metabolically and functionally distinct from omega-3 PUFAs and often have important opposing physiological functions. Either the inefficiency of omega-3 PUFAs or the excess of omega-6 PUFAs will cause many healthy problems. So methods have been sought to increase the amount of omega-3 PUFAs and to improve the omega-6/omega-3 ratio in body. In this study, the sFat-1 gene, which putatively encodes a omega-3 fatty acid desaturase, was chemically synthesized according to the sequence from Caenorhabditis briggssae (with codon usage modified), and constructed into a mammal expression vector pcDNA3. 1-sFat1-EGFP. This vector was introduced into CHO cells by lipid-mediated transfection, and it's expression quickly and effectively elevated the cellular omega-3 PUFAs (from 18-carbon to 22-carbon) contents and dramatically improved the ratio of omega-6/omega-3 PUFAs. Cellular lipids extracts from stably selected cells were analyzed with GC-MS and the results showed that amount of total omega-6 PUFAs dropped from 48.97% (in GFP cells)to 35.29% (in sFat-1 cells), whereas the amount of total omega-3 PUFAs increased from 7.86% to 24.02%, respectively. The omega-6/omega-3 ratio also dropped from 6.23 to 1.47. These data demonstrates the Caenorhabditis briggssae omega-3 Fatty Acid Desaturase gene, sFat-1, was synthesized successfully and can produce omega-3 PUFAs by using the corresponding omega-6 PUFAs as substrates, which shows its potential for use in the production of omega-3 PUFAs in transgenic animals.
Animals ; CHO Cells ; Caenorhabditis ; enzymology ; genetics ; Cricetinae ; Cricetulus ; Fatty Acid Desaturases ; genetics ; physiology ; Fatty Acids ; analysis ; Plasmids ; Polymerase Chain Reaction

OBJECTIVETo investigate the expression of imprinted genes related to Beckwith-Wiedemann syndrome (BWS) in human oocytes and preimplantation embryos for understanding the relationship between assisted reproductive technology (ART) and BWS.

METHODSUsing nested reverse transcription-PCR to analyze the expression of P57KIP2, LIT1, TSSC3 in human oocytes and preimplantation embryos.

RESULTSTranscripts of P57KIP2 were detected in human oocytes and at all stages of preimplantation embryos. LIT1 was expressed only in stages of 8-cell and blastocyst. Transcripts of TSSC3 could not be detected in human oocytes and preimplantation embryos.

CONCLUSIONTranscripts of P57KIP2 and LIT1, imprinted genes related to BWS, were detected in human preimplantation development; ART might affect the epigenetics of imprinted genes in early embryogenesis.

Beckwith-Wiedemann Syndrome ; genetics ; Blastocyst ; metabolism ; Cyclin-Dependent Kinase Inhibitor p57 ; genetics ; Female ; Gene Expression Profiling ; Genomic Imprinting ; genetics ; Humans ; Nuclear Proteins ; genetics ; Oocytes ; metabolism ; Potassium Channels, Voltage-Gated ; genetics ; Pregnancy ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo observe the effect of leukemic cells on blood-brain barrier (BBB) in mice with central nervous system leukemia (CNSL) by establishing mice CNSL model and an in vitro BBB model and explore the mechanism of leukemic cell infiltrating central nervous system (CNS).

METHODSAfter splenectomy, cytoxan intraperitoneal injection, and sublethal irradiation, 10 BALB/c nu/nu mice were transplanted intravenously with 1.2 × 10(7) of SHI-1 human monocytic leukemic cells. Mice were monitored for survival and clinical manifestation of nerve palsy. The leukemic cells engrafted were examined by RT-PCR, histopathology and bone marrow (BM) smears. Immunofluorescence analysis with laser scanning fluorescence confocal microscopy was used to determine the expression of fibrinogen and tight-junction protein ZO-1. An in vitro BBB model composed of human brain microvascular endothelial cells (BMVECs) was developed on a Matrigel-based insert. Different leukemic cell lines were seeded onto the upper compartment of transwell insert. After incubated for 24 h with BMVECs, cells that had migrated into the lower compartment were counted and analyzed.

RESULTS(1) Paralysis with or without sight loss was developed in half the mice 30-35 d after innoculated with SHI-1 cells. Leukemic cells infiltrates were observed in BM and in different part of brain tissues including brain parenchyma. The transcriptions of human MLL/AF6 fusion gene were also detected in BM and brain tissues in paralysis mice. The fibrinogen expression and ZO-1 disruption were detected in the infiltrated tissue. (2) After 24 h incubation with leukemic cells, the BMVECs sheets were disrupted and grew singly and ZO-1 expression was down-regulated markedly. SHI-1 cells showed more injurious to BMVECs and higher invasive rate \[(40.33 ± 1.53)% vs (11.83 ± 1.44)%, P < 0.05\] than HL-60 cells did.

CONCLUSIONOne of the mechanisms of leukemic cells infiltrates CNS in CNSL is injure to the BBB.

Animals ; Blood-Brain Barrier ; physiology ; Central Nervous System ; pathology ; Central Nervous System Neoplasms ; pathology ; HL-60 Cells ; Humans ; Leukemia ; pathology ; Mice ; Mice, Inbred BALB C ; Mice, Nude

OBJECTIVETo evaluate the safety and efficiency of hemi-semi laminectomy approach for the microsurgical treatment of spinal schwannomas.

METHODSA total of 22 patients underwent hemi-semi laminectomy for the microsurgical removal of spinal schwannomas during a period of 2009 and 2011 in Affiliated Hospital of Nantong University. We retrospectively analyzed the clinical outcomes of these patients.

RESULTSOf them, 5 cases were diagnosed with cervical schwannomas, 9 with thoracic schwannomas, and 8 with lumbar schwannomas. All the tumors including two dumbbell schwannomas were totally removed without major complications. Postoperatively, all patients were followed up from 6 to 36 months. The symptoms and signs were obviously improved, and no tumor recurrence or spinal deformity occurred.

CONCLUSIONHemi-semi laminectomy is a safe and effective method for resection of spinal schwannomas.

Adolescent ; Adult ; Aged ; Female ; Humans ; Laminectomy ; methods ; Magnetic Resonance Imaging ; Male ; Microsurgery ; methods ; Middle Aged ; Models, Biological ; Neurilemmoma ; diagnostic imaging ; surgery ; Radiography ; Retrospective Studies ; Spinal Cord Neoplasms ; diagnostic imaging ; surgery ; Treatment Outcome ; Young Adult

OBJECTIVETo study the preparative method of controlled release microspheres incorporating basic fibroblast growth factor (bFGF) and the bioactivities of bFGF, which were released from bFGF microspheres, on the cultured Schwann cells.

METHODSbFGF was microcapsulated with the multiple emulsion encapsulative method using polylactic-co-glycolic acid (PLGA) as coating material. Its morphology, particle size distribution, drug loading, enveloping rate and in vitro release property were studied. The cultured Schwann cells were grouped according to the different ingredients being added to the culture medium of bFGF group or bFGF-PLGA group. Then the cytometry, cytoactivity detection and mitotic cycle analysis of Schwann cells were performed.

RESULTSThe morphology and the particle size distribution of the bFGF-PLGA microspheres were even and good. The drug loading and enveloping rate of microspheres were (27.18 x 10(-3))%+/-(0.51 x 10(-3))% and 66.43%+/-1.24%. The release property of microspheres in vitro was good and the overall release rate was 72.47% in 11 days. The in vitro cellular study showed that: at the first 2 days of plate culture, the cell number and viability of the bFGF group were statistically higher than the bFGF-PLGA group; at the 3rd and 4th days of plate culture, the cell number and viability of bFGF and bFGF-PLGA groups showed no difference; at the 6th and 8th days of the plate culture, the cell number and viability of the bFGF-PLGA group were statistically higher than the bFGF group. By flow cytometry examination, at the 2nd day of plate culture, the G2/M+S percentage of bFGF group was statistically higher than the bFGF-PLGA group, at the 4th and 8th days of plate culture, the G2/M+S percentage of the bFGF-PLGA group was statistically higher than the bFGF group.

CONCLUSIONSIt is practical to prepare the bFGF-PLGA microspheres with the multiple emulsion encapsulative method. bFGF-PLGA microspheres can preserve the bioactivities of bFGF effectively and promote the proliferation of Schwann cells in a long period because of the controlled release of bFGF from the microspheres.

Animals ; Delayed-Action Preparations ; Fibroblast Growth Factor 2 ; administration & dosage ; pharmacology ; In Vitro Techniques ; Microspheres ; Rabbits ; Schwann Cells

BACKGROUNDIncreased signal intensity (ISI) in the spinal cord on T2-weighted MR images has been reported in some previous researches, however no study focused on the position of the ISI in the spinal cord and its potential value. The aim of this study was to investigate the correlation between ISI position and the outcome of surgical treatment for cervical spondylotic myelopathy (CSM) patients.

METHODSA retrospective study was conducted. Pre- and post-operative clinical status was evaluated by modified Japanese Orthopaedic Association (JOA) score. ISI was evaluated according to the T2-weighted sequences. The JOA score and the recovery ratios among patients with ISI in gray matter (group A), in both gray and white matter (group B), and ISI-negative group were compared.

RESULTSTotally 64 patients were enrolled in this retrospective study. Preoperative JOA score of ISI positive and negative group had significant difference, but the recovery ratios had no significant difference (the recovery ratios of the two groups in week 1, week 26, and week 104 were (21.54 +/- 14.65)%, (50.56 +/- 14.76)%, (59.23 +/- 13.08)% and (20.25 +/- 14.32)%, (54.46 +/- 23.16)% and (61.26 +/- 29.4)%, respectively; P > 0.05). The recovery ratios of negative group and group A in week 104 were superior to group B (the recovery ratios of negative group, group A, and group B in week 104 were (61.26 +/- 29.49)%, (65.35 +/- 11.36)%, and (50.33 +/- 10.20)%, respectively; P < 0.05).

CONCLUSIONSPatients with ISI in the gray matter alone on T2-weighted MR images did not have significantly different surgical outcomes compared with those without ISI. Patients with ISI in both gray and white matter had surgical outcomes that were worse than those without ISI.

Female ; Humans ; Magnetic Resonance Imaging ; methods ; Male ; Middle Aged ; Retrospective Studies ; Spinal Cord Diseases ; pathology ; surgery ; Spondylosis ; pathology ; surgery ; Treatment Outcome

OBJECTIVETo investigate the inhibitory effect of small interfering RNA (siRNA) targeting Bax-Bak on the apoptosis of human granulosa cells.

METHODSHuman granulosa cells were transfected with Bax-siRNA and Bak-siRNA either alone or in comibnation, and the cell morphological changes were obsered and the cell apoptosis was detected with flow cytometry. Western blotting was performed to examine the changes in Bax and Bak expressions in the transfected cells.

RESULTSWestern blotting demonstrated significantly weakened expressions of Bax and Bak in the transfected cells. The cell morphology of the cells tranfected with Bak siRNA and with both Bak and Bax siRNA remained normal; the cells with exclusive Bax siRNA transfection presented with basically normal cell morphology, but black spots were noted in the cytoplasm. In the positive and negative control groups, the cells became rounded and shrank with expanded intercellular spaces and numerous black spots in the cytoplasm. Flow cytometry showed apoptotic indexes of 3.44% and 3.97% in cells transfected with Bak siRNA and Bax-Bak siRNA, respectively, significantly lower than that in the negative group. Bax siRNA transfection resulted in an apoptotic index of 19.98%, similar to that in the negative group.

CONCLUSIONInterference of the expression of Bak gene inhibits the apoptosis of human granulosa cells, and the inhibitory effect can be enhanced by simultaneous Bax interference, which, when used alone, does not obviosuly inhibit the apoptosis of human granulosa cells.

Apoptosis ; genetics ; Cells, Cultured ; Female ; Granulosa Cells ; cytology ; Humans ; RNA Interference ; RNA, Small Interfering ; genetics ; Transfection ; bcl-2 Homologous Antagonist-Killer Protein ; genetics ; metabolism ; bcl-2-Associated X Protein ; genetics ; metabolism