Objective:To study the influence of morphine on the expression of nestin in the ependymal epithelia,central gray and hippocampal formations in mice.Methods:Twenty health mice were evenly randomized into control group and experiment group.Mice in the control group were injected with normal saline(0.1 ml daily)and those in the experimental group were injected with morphine (0.1 ml,1 mg daily).Thirty days later,the mice brain samples were harvested and made into paraffin sections.Immumohistochemical ABC technique was used to observe the expression of nestin under light microscope.The images were analyzed with the image analytical system.Results:In the control group,the ependymal epithelia,the central gray,the periventricular gray substances and the hippocampal formations had weak expression of the nestin,with a mean gray scale of 150.98?13.31;there were 5 kinds of nestin-positive cells:(1) the basal cells of ependymal epithelium,(2)cells distributed in the periventricular gray substance and the deep lamella of central gray, (3)cells distributed in the superficial lamella of central gray,the subiculum,the parahippocampal gyrus and the cortex inⅡ,Ⅲlayers of the entorhinal area,(4)cells frequently seen in the rectum of midbrain and the subiculum,and(5)cells distributed in the tectum of midhrain,the hippocampus,gyrus dentatus,parahippocampal gyrus and the cortex in V layer of the entorhinal area;the density of nestin in the subiculum and entorhinal area was(7.20?1.23)mm~2.In the experiment group,the ependymal epithelia,the central gray,the periventricular gray substances and the hippocampal formations had positive expression of the nestin,with the mean gray scale being 133.03?22.28;the density of the above-mentioned 5 kinds of cells increased;the density of nestin in the subiculum and entorhinal area was(10.50?1.43)mm~2.The mean value of gray scale and nestin-positive neurons were significantly different between the 2 groups (P

Hepatocellular carcinoma (HCC) is a common malignant tumor worldwise. The incidence rate of HCC is high and is easy to metastasis and recurrence, which seriously affects human health. Traditional chemical drugs have some challenges such as toxicity, side effects, and multidrug resistance, thus it is urgent to find new drugs and effective targets. Here we synthesized a novel chemical, protonic bis-phenanthroline (H-BP), and the antitumor effect was investigated in the study. The results showed that H-BP could selectively inhibit the proliferation of tumor cells and cause HCC apoptosis. And also, in HCC tumor-bearing mice, H-BP could effectively prevent the growth of tumor mass, even completely eliminate the tumor at medium dose (5 mg·kg^-1) and high dose (10 mg·kg^-1), and meanwhile H-BP has no significant effect on the body weight of mice. The experimental protocol was approved by the Animal Ethics Committee of Southwest University, and the experimental operation was strictly carried out in accordance with the ethical principles of animal use and care. Mechanism studies showed that H-BP induced HCC apoptosis was related to down-regulation the expression of pleomorphic adenoma gene like-2 (PLAGL2), a oncogene transcription factor, resulting in the down-regulation of PLAGL2 downstream proteins hypoxia inducible factor and β-catenin. This study not only introduces the dimerization method to form novel compounds that will provide a new approach for drug design, but also suggests that PLAGL2 may be an effective target in tumor therapy.

"Healthy China" is an important component of Chinese governance system.It is not a simple combination of piecemeal health policies,but a system where multiple organizations and policies perform coordinately and well-functionally.Its key components include health policy system,major participants,institutional arrangement and supporting measures.Medical policies,health promotion policies and social policies that influence population health are three subsystems of health policy system.Government,market (enterprise) and individuals are major participants of building a healthy China.An administrative system that protects health,a well-functioned market economy that promotes health,and a system of social mobilization that encourages individuals to improve health are all essential for "Healthy China".Accordingly,progress in legislation,financing,human resources and technology shoult be in place.Considering its complexity,it suggests that China should take advantage of policy experimentation to find out a feasible way to achieve the goals of "Healthy China".

Objective To develop a method of quantitative analysis of multi-components by single marker (QAMS) for simultaneously determining five compounds in Curcumae Aromaticae Radix.Methods An HPLC method was developed as QAMS to determine curcuma diol,ocathydro-1,4-dihydroxy-1,4-dimethyl-7-(propan-2-ylidene)azulen-5(1H)-one,original curcumol and curcumin in Curcumae Aromaticae Radix,using curdione as intermal reference substance,and the relative correction factor (RCF) of the four components was determined by HPLC with good reproducibility.Their contents in 10 batches of samples,collected from different areas,were determined by both external standard method and QAMS.Result No significant differences were found in the quantitative results of four compounds in 10 batches of Curcumae Aromaticae Radix determined by external standard method and QAMS.Conclusion It is feasible and suitable to evaluate the quality of Curcumae Aromaticae Radix by QAMS.

OBJECTIVETo investigate the distribution of GAD67 and the co-localization with bNOS in the main olfactory bulb of GAD67-GFP knock-in mouse.

METHODSPolymerase chain reaction was applied to identify the genotype of GAD67-GFP knock-in mouse, the animals were sacrificed and frozen sections of olfactory bulb were prepared. The Nissl-staining was performed to show an framework of the neuron in the olfactory bulb. The distribution of GAD67 and co-localization with bNOS were detected by immunofluorescence technique.

RESULTSThe proportion of GAD67-positive cells among DAPI-positive cells were (42.98 ± 0.92)% in glomerular layer, (23.64 ± 0.84)% in mitral cell layer and (77.75 ± 0.84)% in granule cell layer; the bNOS-positive cells mainly existed in glomerular layer and mitral cell layer, very few in granule cell layer. No co-localization of GAD67 and bNOS in granule cell layer and mitral cell layer was found, but there was dispersed distribution in glomerular layer.

CONCLUSIONGAD67-positive neurons mainly appear in glomerular layer and granule cell layer, and the bNOS is mostly expressed in glomerular layer and mitral cell layer; while the co-localization of GAD67 and bNOS only occurs in glomerular layer of olfactory bulb.

Animals ; Gene Knock-In Techniques ; Glutamate Decarboxylase ; genetics ; metabolism ; Green Fluorescent Proteins ; genetics ; metabolism ; Mice ; Mice, Transgenic ; Neurons ; metabolism ; Nitric Oxide Synthase Type I ; metabolism ; Olfactory Bulb ; metabolism ; Tissue Distribution

BACKGROUNDUnfractionated heparin is the most commonly used anticoagulant in continuous renal replacement therapy (CRRT), but it can increase the risk of bleeding. Citrate is a promising substitute. Our study was to assess the efficacy and safety of citrate versus unfractionated heparin in CRRT.

METHODSWe searched the MEDLINE, the EMBASE, the Cochrane Central Register of Controlled Trials, and the China National Knowledge Infrastructure Database until up to November 2011 for randomized controlled trials comparing citrate with unfractionated heparin in adult patients with acute kidney injury prescribed CRRT. The primary outcome was mortality and the secondary outcomes included circuit survival, control of uremia, risk of bleeding, transfusion rates, acid-base statuses, and disturbance of sodium and calcium homeostasis.

RESULTSFour trials met the inclusion criteria. Meta-analysis found no significant difference between two anticoagulants on mortality. Less bleeding and more hypocalcemic episodes were with citrate. Citrate was superior or comparable to unfractionated heparin in circuit life.

CONCLUSIONSCitrate anticoagulation in CRRT seems to be superior in reducing bleeding risk and with a longer or similar circuit life, although there is more metabolic derangement. Mortality superiority has not been approved.

Anticoagulants ; therapeutic use ; Citric Acid ; therapeutic use ; Heparin ; therapeutic use ; Humans ; Randomized Controlled Trials as Topic ; Renal Replacement Therapy ; methods

OBJECTIVETo study the histochemical staining in the diagnosis of osteosarcoma.

METHODSTo compare the effectiveness of picrosirius red, improved Ponceau trichrome and Masson trichrome staining methods on bone formation tissues in conventional osteosarcoma, paraosteal osteosarcoma, periosteal osteosarcoma, extraskeletal osteosarcoma, inflammatory myofibroblastic tumour, malignant fibrohistiocytoma, chondrosarcoma, fibrosis with ossification and calcification.

RESULTSWith modified Ponceau trichrome staining, bone formation tissues showed a homogenous, orange-red interblended with blue in color. From osteoid to mature bone the color changed from orange-red, light blue to dark blue. Fibrotic tissue was stained blue in color with striated appearance. Cartilage was not stained. Picrosirius red method gave bone formation tissues homogenous staining. Along with bone maturation, from osteoid tissue to mineralized bones, the color showed changes from light red, yellow, orange-red, red to dark purple. The cartilage demonstrated homogenous light red in color. Fibrous tissue stained red interblended with yellow in color, striated in shape. With Masson trichrome staining osteoid displayed pale blue and mineralized bone showed dark blue in color. Fibrotic tissue showed a striated blue staining.

CONCLUSIONThe modified Ponceau trichrome and Picrosirius red staining methods are better than Masson trichrome to demonstrate bone formation tissue in osteosarcoma. The former two methods could be also used in study on bone formation.

Bone Neoplasms ; pathology ; Histocytochemistry ; Humans ; Osteosarcoma ; pathology ; Staining and Labeling ; methods

OBJECTIVERett syndrome (RTT) is a severe childhood neurodevelopmental disorder mainly affecting females. The pathogenic gene is located at Xq28, which codes for the methyl-CpG-binding protein 2. MECP2 gene is affected by X chromosome inactivation (XCI). The different XCI patterns of females could affect the expression ratios of pathogenic gene, causing changes in clinical symptoms. In order to understand the XCI patterns in RTT patients and the relationship between XCI pattern, genotype and phenotype, the XCI patterns in patients with RTT and their mothers, the parental origin of the priority inactive X chromosome in RTT, and the relations of XCI patterns with genotype and phenotype in RTT cases were analyzed.

METHODSGenomic DNA was extracted from peripheral blood of 55 cases with RTT (52 with MECP2 mutations, 3 without mutations), 53 mothers of RTT cases and 48 normal female controls. DNA was digested with methylation sensitive restriction endonuclease Hpa II. Then the undigested and digested DNAs were amplified via PCR for the first exon of human androgen receptor (AR) gene. PCR products were analyzed by Genescan.

RESULTSThe heterozygotic rates of AR gene were 82%, 77% and 83% in RTT patients, mothers and controls, respectively. XCI distribution pattern of RTT was different from that of the mothers and control, P < 0.05. More mothers and controls than RTT patients were in the area of XCI 50:50 - 59:41. The differences between them were statistically significant (P < 0.05). No significant difference in XCI distribution patterns between mothers and the control groups was found (P > 0.05). Non-random XCI rates in the areas of XCI >or= 65:35 and >or= 80:20 were 53.35% and 17.8%, respectively, in RTT patients, compared with the mothers group (36.6%, 7.3%) and control group (35%, 10%), it was higher in RTT patients, but the difference was not statistically significant (P > 0.05). In 18 of 21 cases with XCI >or= 65:35, the priority inactive X chromosome was of paternal origin (85.7%). Variable XCI patterns were observed in the same gene mutation patients. The highly skewed XCI as well as the random XCI were found in patients with mild, severe and typical phenotype. The rate of highly skewed XCI in atypical patients was higher than that in typical RTT patients. The rate of highly skewed XCI in T158M was higher than the other type mutations. No highly skewed XCI was observed in cases with R133C mutation.

CONCLUSIONThe XCI distribution pattern of RTT patients was different from that of RTT mother and control groups. There was no significant difference in XCI distribution patterns between mothers and the control groups. It was not a main genetic pattern in RTT that mothers as the carriers to transmit the pathogenic gene to the patients. Non-random XCI was not the main XCI pattern in RTT patients. The priority inactive X chromosome was mainly of paternal origin. XCI could modify the clinical phenotype of RTT, but had limitations in explaining all the phenotypes manifested in RTT cases.

Adolescent ; Adult ; Child ; Child, Preschool ; Chromosomes, Human, X ; genetics ; Female ; Genetic Predisposition to Disease ; Genotype ; Heterozygote ; Humans ; Mothers ; Mutation ; Phenotype ; Receptors, Androgen ; genetics ; Rett Syndrome ; diagnosis ; genetics ; X Chromosome Inactivation ; genetics

OBJECTIVETo investigate the expression level of inhibitor of apoptosis protein survivin gene in human brain glioma and its role in malignant proliferation and antiapoptosis of brain glioma.

METHODSEighty-three cases of brain glioma specimen was chosen, protein expression of survivin and proliferating cell nuclear antigen (PCNA) was investigated by immunohistochemistry streptavidin-biotin complex (SABC) method, the immunoreactivity score (IRS) of survivin and the proliferative index (PI) were counted. Apoptotic cells were screened by TdT-mediated dUTP-biotin nick-end labeling (TUNEL) method, and the apoptotic index (AI) of brain glioma was calculated.

RESULTSThe survivin IRS, PI and AI of brain glioma were 3.8 +/- 3.9, (28.4 +/- 19.5)% and (1.0 +/- 0.8)% respectively, and all of them were elevated with the increase of pathological grade of brain glioma (P < 0.01 for all). PI in survivin positive group was significantly higher than that in survivin negative group (P < 0.01), and PI was positively correlated with survivin IRS (r = 0.740, P < 0.01). There was no significant difference between AI in survivin positive group and that in survivin negative group (P > 0.05), however, AI was negatively correlated with survivin IRS (r = -0.307, P < 0.01).

CONCLUSIONSSurvivin is overexpressed in brain glioma, and which may play important roles in malignant proliferation and antiapoptosis of brain glioma.

Adolescent ; Adult ; Aged ; Apoptosis ; Brain Neoplasms ; genetics ; metabolism ; pathology ; Cell Proliferation ; Child ; Child, Preschool ; Female ; Glioma ; genetics ; metabolism ; pathology ; Humans ; Immunohistochemistry ; In Situ Nick-End Labeling ; Inhibitor of Apoptosis Proteins ; Male ; Microtubule-Associated Proteins ; biosynthesis ; genetics ; Middle Aged ; Neoplasm Proteins ; biosynthesis ; genetics ; Proliferating Cell Nuclear Antigen ; biosynthesis

OBJECTIVETo investigate telomerase activity of MCF-7 mammary cancer cells during apoptosis induced by sodium butyrate (SB) in vitro and its mechanism.

METHODSThe proliferative activity of MCF-7 cells was assessed by morphology and MTT assay. Cell apoptosis was confirmed by DNA fragmentation and phosphatidylserine (PS) externalization. Telomerase activity was examined by TRAP-ELISA. The expression status of telomerase subunits was analyzed by RT-PCR.

RESULTSA time- and dose-dependent inhibition was detected in MCF-7 cells treated with SB. At 72 hr after SB (2.5 mmol/L) treatment, MCF-7 cells were apoptotic with a rate of 84.3% by flow cytometric assay (AnnexinV/PI double staining). Apoptosis was also confirmed by DNA fragmentation. Telomerase activity and expression level of hTERT, the key subunit of telomerase, decreased at 24-hour time point after SB treatment. No significant changes were observed in the expression of hTR and hTP, the other two subunits of telomerase.

CONCLUSIONTelomerase activity decreases in MCF-7 cells during apoptosis induced by sodium butyrate. The underlying mechanism might be related to the down regulation of hTERT transcription.

Apoptosis ; drug effects ; Breast Neoplasms ; enzymology ; pathology ; Butyrates ; administration & dosage ; pharmacology ; Cell Line, Tumor ; DNA-Binding Proteins ; biosynthesis ; genetics ; Dose-Response Relationship, Drug ; Humans ; RNA, Messenger ; biosynthesis ; genetics ; Telomerase ; biosynthesis ; genetics ; metabolism ; Time Factors