Objective:To investigate the differences in related indices of ulcerative colitis (UC) respectively induced by free drinking and intragastric administration of dextran sodium sulfate (DSS) in mice to provide experimental reference for the optimization of UC model.Methods:Totally 30 C57BL/6 mice were randomly divided into the normal control group,free drinking group and intragastric administration group with 10 ones in each.The mice drank water freely with free drinking or intragastric administration of 3% DSS solution at the dose of 4 g·kg-1·day-1 for 7 days to establish the UC model.The differences in disease activity index (DAI),histological damage sore and activity of myeloperoxidase (MPO) among the groups were compared.Results:Two mice died during the experiment in the free drinking group,and DAI of survival mice was (8.8±1.6).There was no death of mice in intragastric administration group,and DAI was (9.0±0.8),and there was no significant difference in DAI between the groups (P>0.05),while the coefficient of variation in the free drinking group was higher than that in the intragastric administration group (18.7 vs 8.6).The colonic histological damage score of the free drinking group and the intragastric administration group was 24.8±4.2 and 27.0±2.8,respectively,which was typical inflammatory change with no significant difference (P>0.05),while the coefficient of variation of the free drinking group was higher than that of the intragastric administration group (16.9 vs 10.4).MPO of the normal control group,free drinking group and intragastric administration group was (0.41±0.03),(2.32±0.34) and (2.05±0.18) U·g-1,respectively.Compared with the normal control group,significant difference in MPO was shown in the free drinking group and the intragastric administration group (P<0.01),while there was no significant difference in MPO between the groups (P>0.05),and the coefficient of variation in the free drinking group was higher than that in the intragastric administration group (14.7 vs 8.8).Conclusion:Both free drinking and intragastric administration of DSS can successfully induce the UC model in mice.Compared with the free drinking group,the intragastric administration group has low mortality rate and low coefficient of variation.Therefore,intragastric administration has more advantages than free drinking in inducing the UC model in mice.

Objective To analyze the amount and allocation of pediatric healthcare resources in Chengdu,and to recommend on local pediatric healthcare resources shortages.Methods Pediatric healthcare resources data of Chengdu came from pediatric relevant data reported regularly at yearend by the counties and districts to healthcare administration of the city.Data from such reports were subject to statistics with various indexes and descriptive analysis.Analyzed in focus was the distribution in 2015 of pediatric healthcare resources of the city among medical institutions of various types,levels and properties,as well as causes for such shortage.Results Tertiary hospitals hold 62.3% of the pediatric beds and 64.2% of pediatricians,and provide around 70% of the medical workload for pediatric outpatients and inpatients,upon the majority of pediatricians with master degree and above,and senior ones of/above associate chief physician titles;Tertiary hospitals have 58.7% of the pediatric beds and 49.7% of pediatricians,yet the outpatients served by specialized hospitals were 5% above tertiary hospitals.Conclusions The imbalance and shortage in total of pediatric resources in Chengdu result from stable source of manpower supply,high professional risk exposure,low income,and long training duration among others.Such measures as a better pediatrician development system,greater incentives for pediatric development,and enhanced development pediatric service consortiums,as well as greater support for private specialized pediatric hospitals.Those measures combined can effectively alleviate the shortage of pediatric resources.

Aged ; Angioplasty, Balloon ; Arterial Occlusive Diseases ; diagnostic imaging ; physiopathology ; therapy ; Cerebral Arterial Diseases ; diagnostic imaging ; physiopathology ; therapy ; Cerebral Arteries ; diagnostic imaging ; Female ; Follow-Up Studies ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Renal Artery ; diagnostic imaging ; Renal Artery Obstruction ; diagnostic imaging ; physiopathology ; therapy ; Risk Factors ; Stents ; Tomography, X-Ray Computed

OBJECTIVETo develop an effective method for screening recombinant hairpin RNA expression plasmids using single restriction endonuclease analysis.

METHODSThe double-strand DNA fragment containing a ClaI site (the flanking sequences of which were not complementary) was annealed and ligated into small hairpin RNA (shRNA) expression vector pSilencer-4.1 that did not contain ClaI site to construct the circular pSilencer-4.1-ClaI vector. With BamHI and HindIII, the pSilencer-4.1-ClaIwas digested and ligated with the DNA template of green fluorescence protein (GFP) shRNA that did not include a ClaI site. The plasmid DNA of the positive clones was extracted and digested with ClaI, and the inserted DNA sequence of the non-linearized plasmid was identified by sequence analysis.

RESULT AND CONCLUSIONDNA sequencing showed that pSilencer-4.1-ClaI was correctly constructed and the plasmids resistant to ClaI digestion were all recombinant vectors encoding GFP shRNA. The constructed pSilencer-4.1-ClaI can be used as a universal vector to construct the shRNA expression plasmid, and the incorporated ClaI sites may allow efficient screening of recombinant shRNA expression vectors.

Base Sequence ; Gene Expression ; Genetic Engineering ; methods ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; genetics ; Inverted Repeat Sequences ; Molecular Sequence Data ; Plasmids ; genetics ; RNA, Small Interfering ; genetics ; Restriction Mapping ; methods ; Sequence Analysis, DNA ; Time Factors

OBJECTIVETo investigate the phenotypes and functions of cord blood dendritic cells of fetuses whose mothers are patients with chronic hepatitis B.

METHODSPeripheral blood and cord blood mononuclear cells (PBMC) were isolated from whole blood by density gradient centrifugation with Ficoll-Hypaque. The adherent cells were cultured in AIM-V medium containing recombinant human IL-4, TNF-alpha and GM-CSF. On day 9, mature DCs (mDC) were harvested and used for phenotype analysis. The amounts of IL-12 which dendritic cells produced were measured. The dendritic cells that were studied and compared were from cord blood of fetuses of both CHB positive and negative mothers and from CHC adult peripheral blood.

RESULTSThe expression rate of CD80 and CD83 of chronic hepatitis B mother cord blood dendritic cells was low compared with that of the healthy cord blood, healthy adult peripheral blood, and chronic hepatitis B adult peripheral blood, P < 0.05. The amount of IL-12 produced by chronic hepatitis B mother cord blood dendritic cells was lower than that of healthy cord blood, healthy adult peripheral blood, chronic hepatitis B adult peripheral blood (P < 0.05). The T lymphocyte proliferation inducing ability of dendritic cells of healthy adult peripheral blood was higher in inducing cord blood T lymphocytes proliferation, which was greater than that of the healthy adult peripheral blood in inducing adult T lymphocytes and was greater than that of the healthy cord blood dendritic cells in inducing cord blood T lymphocytes, which was greater than that of the healthy cord blood in inducing adult T lymphocytes, which was greater than that of chronic hepatitis B mothers in inducing cord blood T lymphocytes, which was greater than that of chronic hepatitis B mother cord blood in inducing adult T lymphocytes.

CONCLUSIONThe maturation and functioning of CHB mother cord blood dendritic cells were lower than those of healthy cord blood, healthy adult peripheral blood and CHB adult peripheral blood.

Adult ; Cells, Cultured ; Dendritic Cells ; cytology ; immunology ; Female ; Fetal Blood ; immunology ; Hepatitis B, Chronic ; immunology ; Humans ; Phenotype ; Pregnancy ; Pregnancy Complications, Infectious ; immunology ; T-Lymphocytes ; immunology

Objective To compare the efficacy of transplanting bone marrow mesenchymal stem cell (BMSC) or microenvironmental induced BMSC ( iBMSC) into the ischemic myocardium of rats with myocardial infarction. Methods iBMSC was defined as BMSC co-cultured with myocardial cells for 2 weeks. The stem cells or equal volume PBS were injected into ischemic border zone 1 wk after experimental infarction. Cardiac performance was evaluated at 1, 2, and 4 wk after cell transplantation by echocardiography and analyzed histologically at 4 wk after cell transplantations. Results Compared with PBS group, both BMSC and iBMSC transplantations reduced infarct size. iBMSC enhanced the beneficial effects of BMSC on improving cardiac function (FS: 28.5% ±4.3% in PBS, 29.0% ±2.0% in BMSC and 45. 1% ±3. 1% in iBMSC group at 4 weeks post transplantation, iBMSC group vs. PBS group P <0. 05, iBMSC group vs. BMSC group P <0. 05). Immunofluorescence microscopy results revealed co-localization of SPIO-labeled transplanted cells with cardiac markers for cardiomyocytes, indicating regeneration of damaged myocardium. Conclusion Our data suggest that iBMSC implantation is more effective on improving cardiac function than BMSC implantation in this model. iBMSC might serve as a new promising therapeutic cell source for regenerating ischemic myocardium in patients with post-infarction heart failure.

OBJECTIVETo construct an eukaryotic expression vector for miRNA-1-2 that can be expressed in rat H9C2 cardiomyocytes.

METHODSThe precursor miRNA (pre-miRNA) DNA template for miRNA-1-2 was designed and generated by PCR amplification. The DNA template was inserted into the hairpin RNA expression vector pSilence-4.1-neo and identified by DNA sequencing analysis. The recombinant plasmid DNA was then transfected into H9C2 cells via Lipofectamine, and the green fluorescence protein expression vector pEGFP-N3 served as the transfection marker. Twenty-four hours after transfection, the total cellular RNA was extracted using TRIzol reagent, and thermoscript reverse transcriptase (RT)-PCR was performed to determine miRNA-1-2 precursor expression.

RESULTS AND CONCLUSIONDNA sequencing indicated that the miR-1-2 expression plasmid was correctly constructed. The precursor miRNA-1-2 was successfully expressed in the H9C2 cells, and the expression of Hand2 protein could be efficiently inhibited by miRNA-1.

Animals ; Basic Helix-Loop-Helix Transcription Factors ; genetics ; metabolism ; Cell Line ; Down-Regulation ; Green Fluorescent Proteins ; genetics ; metabolism ; MicroRNAs ; genetics ; Myocytes, Cardiac ; cytology ; metabolism ; Plasmids ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; RNA, Small Interfering ; genetics ; Rats ; Transfection

Objective To investigate the status of endemic fluorosis in Boxing County in Shandong Province at present,and to provide the scientific evidence for making strategies in prevention and control.Methods Children aged 8-12 years old and adults above 30 years old were selected from 8 endemic fluorosis villages in 2 fluorosis of children aged 8-12 years old were diagnosed by Dean method and skeletal fluomsis diagnosed by clinic and X-Rays.Results Eight villages in 2 towns were chosen underwent epidemiological investigation.Eight villages had water fluoride content＞4.50 mg/L.the highest water fluoride content was 5.78 mg/L.The total rate of dental fluorosis of children aged 8-12 years old WaS 90.70%(195/215),the index of dental fluorosis was 2.15 and the rate of dental damage was 24.65%(53/215).The rate of skeletal fluorosis detected by clinic and X-rays in adults older than 30 years old were 30.71%(78/254)and 16.54%(42/254),respectively.The averaged fuoride level in urine wa8 over 1.50 mg/L in 98.95%(189/191)of children aged 8-12 years old and in 97.92%(235/240)adults older than 30 years old,with the highest respectively being 14.50 mg/L and 17.99 mg/L.Conclusions In Boxing County in Shandong Province,endemic fluorosis is not effectively controlled.So endemic fluorosis control mfist be strengthened.

Objective To investigate the ability of human bone marrow mesenchymal stem cells (hBMSCs), eocultured with semi-permeable membrane separated neonatal rat ventricular myocytes, to differentiate into cardiomyocytes.Methods hBMSCs were isolated and purified by density gradient centrifugation and adherence screening method.cells were expanded as undifferentiated cells in culture for more than 3 passages and their phenotypes were identified with flow cytometer, hBMSCs were cocuhured with neonatal rat ventricular myocytes in a rate of 1:10 separated by semi-permeable membrane.GATA4 mRNA was detected by RT-PCR; Immunocytochemistry, and Immunostaining were used to detect sarcomeric α-actinin, desmin, cTnT, and cTnI protein level.Results CD29 (98.64% ± 0.80%) and CD44 (96.70% ± 1.50% ) were the major surface markers of hBMSCs.After coculturing with semi-permeable membrane separated neonatal rat ventricular myocytes, the first contraction of single cells was noted at day 7 and GATA4 expression was detected on these cells by RT-PCR after 1 to 3 weeks coculture.Desmin, sacomefic ±-actinin, cTnI and cTnT could be detected by immunocytochemistry and immunostaining on some of these cells.conclusion hBMSCs possess the potential to differentiate into myocardial cell phenotype in the cardiac microenvironment.Direct contact with cardiomyocytes was not necessary required for hBMSCs differentiation.

OBJECTIVEThis study aims to establish and evaluate the methodology of isolated rabbit eye (IRE) test.

METHODSIRE test was performed according to modifications of the in vitro toxicology (INVITTOX) Protocol No.85: Rabbit enucleated eye test by European Centre for the Validation of Alternative Methods (ECVAM), and then 26 chemicals and 26 cosmetic products were tested in both in vitro IRE and in vivo Draize tests. A statistical analysis was conducted to determine the relevance of the IRE test to the data generated in the Draize test.

RESULTSIRE test was established successfully in our laboratory. It was shown that ranking correlation and class concordance were fairly well between the IRE test and the Draize test for 26 reference chemicals (Fisher's Exact Test χ(2)=51.314, P<0.001; McNemar P=0.261; Gamma=0.960, P<0.001; Kappa=0.843, P<0.001) and 26 cosmetic products (Fisher's Exact Test χ(2)=15.522, P<0.001; McNemar P=0.311; Gamma=0.967, P<0.001; Kappa=0.611, P<0.001).

CONCLUSIONIRE test was established successfully for in vitro testing of eye irritation as an alternative to Draize test.

Animal Testing Alternatives ; Animals ; Cosmetics ; toxicity ; Eye ; drug effects ; Irritants ; toxicity ; Rabbits ; Toxicity Tests ; methods