Objectives: To investigate the effect of butyrate, the by-products of diet fiber fermented in colon, on the apoptosis of human colon cancer cell and its mechanism for colon cancer prevention. Methods: The subculture of human colon cancer cell line SW1116 was cultured in medium without or with different concentration of butyrate(2、3、4、7 and 10 mmol/L). The cell proliferation rate and apoptosis were measured by MTT and flow cytometer or electro-microscopy respectively after 6、24、48 and 72 cultivation. Results: By the increasing of butyrate concentration and cell culture time, the proliferation of SW1116 cell was inhibited gradually with a highest rate of 59.19% during the study period. The percent of G 1 cell was increased gradually whereas the percent of S cell was decreased. The apoptosis rate of SW1116 was increased gradually with a highest rate of 30.62% at the same time. The ultrastructure of SW1116 cell was changed after butyrate was added to culture medium. Conclusions: Butyrate could not only inhibit the proliferation of colon cancer cell line but also induce it apoptosis.

BACKGROUND: Previous studies have demonstrated that neural stem cells play potential therapeutic effects on the repair of spinal cord injury. However, the time for acquiring the best allotransplantation effects remains unclear.OBJECTIVE: This study was designed to observe the repairing effects of allotransplantation of embryonic neural stem cells on the motor function of rat two posterior limbs after spinal cord injury and investigate the time effectiveness of the allotransplantation.DESIGN: A controlled observational experiment.SETTING: Laboratory of Molecular Biology, Dalian Medical University; Laboratory of Biomedicine, School of Environmental and Biological Science and Technology, Dalian University of Technology, Dalian, Liaoning Province, China.METHODS: This study was performed at the Laboratory of Molecular Biology, Dalian Medical University & Laboratory of Biomedicine, School of Environmental and Biological Science and Technology, Dalian University of Technology between July and August 2003. One albino rat of gestational 14-16 days was sacrificed for harvesting embryonic rat brain cells. Embryonic rat cerebral cortex and subcortical periventricular brain tissue were taken for in vitro culture of rat embryonic neural stem cells. An additional 30 adult Sprague Dawley rats were randomly divided into 3 groups with 10 rats in each group: control, early allotransplantation and delayed allotransplantation groups. All 30 rats were subjected to spinal cord transection injury, leading to rat paralysis of both lower extremities. Embryonic rat neural stem cells were transplanted into the rats in the early and delayed transplantation groups at 3 days and 3 weeks after injury, respectively. Following allotransplantation, motor function of rat two lower extremities was followed. At 4 weeks after allotransplantation of neural stem cells, rat spinal cord was harvested from transplanted region for immunohistochemistry in order to observe and compare the morphological change of rat spinal cord tissue among the 3 groups. The following protocol was performed in accordance with ethical guidelines stated in Guide for the use and care of laboratory animals, approved by the Committee on the Care and Use of Laboratory Animals of the Institute of Laboratory Animal Resources Commission on Life Scineces, National Research Council, China (1985).MAIN OUTCOME MEASURES: Motor functional recovery of rat two lower extremities after neural stem cell transplantation. Histomorphological change of rat spinal cord at 4 weeks after neural stem cell transplantation.RESULTS: Thirty rats were included in the final analysis. In the early and delayed transplantation groups, the motor function of rat two lower extremities was noticeably improved, in particular in the early transplantation group. In the two experimental groups, muscular strength of paralyzed rat two lower extremities began to recover 5 or 6 days after transplantation of neural stem cells. Two or three weeks later, all rats in the two experimental groups could crawl and four weeks later, two extremities could move actively (approximately approaching to score 3 prescribed as follows). In the control group, no recovery of paralyzed extremities was found. At 4 weeks after transplantation, in the early transplantation group, proliferative tissue could be visible in the spinal cord transplantation region. Through the use of microscope, a considerable number of new cells were found that presented with neuronal and glial cell-positive staining. In the control group, a cavity between two broken ends could be visible. Meanwhile, necrosis and vacuolar degeneration, and other symptoms in the stump of spinal cord were observed with a microscope. In the delayed transplantation group, the histomorphological change of spinal cord region was between the other two groups. No typical histomorphological change was found. A number of new cells were apparent with a microscope, but the number was less compared with the early transplantation group.CONCLUSION: Allotransplantation of embryonic neural stem cells promotes the recovery of rat motor function after spinal cord transection. Early transplantation acquires better therapeutic effects.

BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) are easily isolated and amplified, and facilitate the exogenous gene transfer and expression. In the human medicine, it is believed that BMSCs are ideal therapeutic cells and target cells in the gene therapy.OBJECTIVE: To investigate liposome-mediated cytosine deaminase (CD) gene transfecting rabbit BMSCs and its gene expression. DESIGN: A single sample observation. SETTING: Dalian Research and Development Center for Stem Cell and Tissue Engineering; Department of Biochemistry, College of Basic Medical Science, Dalian Medical University.MATERIALS: This study was performed at in the Dalian Research and Development Center for Stem Cell and Tissue Engineering; Department of Biochemistry, College of Basic Medical Science, Dalian Medical University from March 2006 to June 2007. New Zealand big-ear white rabbits of either gender, weighing 2.0-2.5 kg, with the age of 5 months old, were included in this study. METHODS: The CD gene was obtained from E.coli JM109 DNA by polymerase chain reaction (PCR). The fragment was cloned into pMD19-T vector. After restriction enzyme BamHI/XhoI digestion analysis and DNA sequence analysis, pIRES2-AcGFP1-CD eukaryotic expression plasmid was constructed. Meanwhile, BMSCs were harvested, cultured and identified. After enzyme digestion of eukaryotic expression plasmid, the rabbit BMSCs were transfected by Lipofectamine 2000-mediated method. Twenty-four hours after transfection, expression of green fluorescent protein was observed under an inverted fluorescent microscope. MAIN OUTCOME MEASURES: Construction of eukaryotic expression plasmid and identification of CD gene-transferred BMSCs. RESULTS: CD gene was cloned and connected to eukaryotic expression plasmid with green fluorescence. Twenty-four hours after transfecting rabbit BMSCs, it was found under an inverted microscope that under the excitation of 488 nm blue light, green fluorescence appeared in the pIRES2-AcGFP1-CD and pIRES2-AcGFP1 empty-plasmid transfected BMSCs, but not in the non-transfected ones. It indicates that CD gene successfully transferred BMSCs. CONCLUSION: BMSCs are ideal vectors in the CD gene therapy.

Objective To identify the expression level of miR-125a-5p in colorectal cancer (CRC) with various differentiation,and to investigate its diagnostic significance.Methods Real-time polymerase chain reaction was used to analyze the expression level of miR-125a-5p in tumors and paired normal tissues of different differentiated CRC.Receiver-operating-characteristic (ROC) curve was performed to evaluate the specificity and sensitivity of using miR-125a-5p as biomarkers to discriminate CRC patients from normal persons.Results Compared with normal tissues,miR-125a-5p was down-regulated in both high and moderate differentiation,miR-125a-5p could distinguish CRC from normal persons with high sensitivity and specificity,which were 80.00 ％ and 90.00 ％,respectively.Conclusions miR-125a-5p down-regulated in CRC indicates that miR-125a-5p contributes to tumorigenesis,and may be a potential biomarker for CRC.

Objective To establish effective method for large-scale purification of islet cells from pig pan-cress. Methods Pig pancreas tissue was digested with collagenase P followed by purification in a HCA-Fi-coil dis continuous gradient using Cobe2991 cell separator. After isolation, the islet cell yield and purity were evaluated with light microscope with DTZ staining, and the islet function assessed by insulin release as-say in vitro. Results The number of the islets coll ected from each pancreas averaged (275 000±20 895)islet equivalents (IEQ) before purification, and (230 350±26 679) IEQ after the purification with discon-tinuous gradient centrifugation. From each gram of the pancreatic tissue, (2710±229) IEQ were obtained with an average purity of (50.2±1.95) %. The purified islets responded well to high-concentration (16.7 mmol/L) glucose stimulation with a 4. 74-fold increase of insulin secretion over the basal level (3.3 mmol/L, P <0.001). Conclusion The established method can be applicable for large-scale purifi-cation of fully functional islet cells from pig pancreas.

OBJECTIVETo identify dysregulated microRNA(miRNA) that can act as novel biomarker for colorectal cancer screening.

METHODSReal time-polymerase chain reaction was used to detect the expression profile of miRNA in tumor and paired normal tissues, and the significant dysregulated miRNA was examined by non-paired t-test. Receiver operating characteristic(ROC) curve was performed to evaluate the specificity and sensitivity of miR-363 and miR-490-5p as biomarkers to discriminate colorectal cancer patients from normal person.

RESULTSSeven-three dysregulated miRNAs were found in male samples, of which 6 miRNAs were up-regulated and other 67 miRNAs were down-regulated, while 42 dysregulated miRNAs were found in female samples, of which 5 were up-regulated and 37 were down-regulated in tumor tissues compared with normal tissues. Among above dysregulated miRNAs, 33 miRNAs had significant differences, besides, dysregulated expression level of 10 of 33 miRNAs was over 5 folds in male and female as well as mixed samples.

CONCLUSIONSThe expression pattern in female is different from that in male. miR-363 and miR-490-5p possess the potential in screening colorectal cancer patients from healthy people.

Adult ; Aged ; Aged, 80 and over ; Biomarkers, Tumor ; analysis ; Case-Control Studies ; Colorectal Neoplasms ; diagnosis ; Early Detection of Cancer ; Female ; Humans ; Male ; MicroRNAs ; analysis ; Middle Aged ; Sensitivity and Specificity ; Young Adult

Objective To investigate the effect of remimazolam on anesthetic effect and postoperative cognitive function during painless bronchoscopy in elderly patients.Methods A total of 90 patients with painless bronchoscopy admitted to Wenzhou People's Hospital were selected.And randomly dividing into 45 patients in control group,45 patients in observation group.The control group was given propofol,and the observation group was given remimazolam.Mean arterial pressure(MAP),heart rate(HR),and blood oxygen saturation(SpO2)were monitored.Effective time after induction,recovery time after withdrawal and discharge time were compared.Patient's cognitive function was assessed using the mini-mental state examination(MMSE),auditory word learning test(AVLT),shape connection test(STT),and animal language fluency test(AFT).Enzyme linked immunosorbent assay(ELISA)was used to determine substance P(SP),C-reactive protein(CRP),noradrenaline(NE),tumor necrosis factor-α(TNF-α)in serum,interleukin(IL)-6 and prostaglandin E2(PGE2)concentrations.Results Compared with the control group,the MAP of the observation group patients at time points T2 and T3 showed statistically significant differences(P<0.05);HR was especially significant at T2,T3 and T6 periods(P<0.05).SpO2 at T2 and T3(P<0.05).The awakening time,discharge time of the observation were shorter than control(P<0.05).At 1 day after surgery,the MMSE score,STT score,AFT score were significantly higher(P<0.05);The serum levels of SP,PGE2 and NE and IL-6,TNF-α and CRP were decreased in the control(P<0.05).The incidence of adverse reactions was 17.78％in control group and 8.89％in observation group(χ2=7.654,P=0.031).Conclusion Remazolam is used in painless bronchoscopy in elderly patients,intraoperative hemodynamics is stable,having little impact on postoperative cognitive function,and inhibits the release of inflammatory factors and the secretion of pain mediators,which is worthy of clinical use.