Background Recently,there were many studies on corneal innervations during mammalian development.However,there were fewer studies on discussing corneal innervations before and after mouse eye openings.Objective The present study was to investigate the change in the regulation of corneal nerve fiber length and density before and after mouse eye openings to offer a basis for clinical research in human.Methods Thirty SPF C57BL/6 mice were divided into postnatal 1 day(P1 d),P7 d,P13 d(1 day before eye opening),P14 d(eye halfopened),P17 d(1 day after eye opening)and P23 d(7 day after eye opening)groups,with 5 mice and 10 eyes for each group.Entire corneal stretches were prepared and immunostaining with an anti-neuron-specific β-Ⅲ tubulin antibody was performed to label the corneal nerve fibers.Confocal microscopic pictures from the corneal dorsal-nasal region (DN),dorsal-temporal(DT),ventral-nasal region(VN)and ventral-temporal(VT)were taken using Delta Vision Core.From these pictures,the mouse corneal area,total length and density of nerve fibers in the 4 regions were calculated.The use of the animals complied with Statement of ARVO.Results Corneal areas of P1 d,P7d,P13 d,P14 d,P17 d and P23 d mice were(0.404±0.007),(1.362±0.154),(1.573±0.080),(1.603±0.046),(1.847±0.052),(2.445±0.798)mm2,respectively ; the total lengths of nerve fibers were(3.718±1.044),(19.065±3.350),(23.687±0.907),(27.309±2.477),(31.989±3.976),(41.214±1.573)mm,respectively ; the densities of nerve fibers were(9.592±1.138),(14.506±1.908),(15.088±1.241),(16.772±1.897),(16.821±2.102),(17.660±1.216)mm/mm2,respectively,all showing significant increases with age(F =22.906,P =0.000 ; F =0.424,P =0.000 ; F =2.375,P=0.000).A positive correlation of the increasing corneal areas and increasing lengths of nerve fibers was found(r=0.983,P＜0.01).Nerve fiber densities in the four corneal regions significantly increased with age(DN region:F =0.159,P =0.000 ; DT region:F =2.1 72,P =0.001 ; VN region:F =1.998,P =0.000 ; VT region:F=2.352,P=0.000).From P13 d to P14 d,the corneal nerve fiber densities in the DN region decreased by 6.0％ without significant difference(t =0.589,P =0.572); and the corneal nerve fiber densities in the DT region,VN region and VT region decreased by 4.6％,5.5％ and 0.1％,respectively,without significant difference from P14 d to P17 d(t=0.549,P=0.596;t=0.701,P=0.501 ;t=-0.100,P=0.919).Conclusions The development of nerve fibers in the whole cornea or the four corneal regions is influenced by eye opening in mouse to various extents.From P13 d to P14 d,the corneal nerve fiber densities in the DN region decreased by 6.0％ without significant difference.From P14 d to P17 d,the corneal nerve fiber densities in the DT region,VN region and VT region decrease by 4.6％,5.5％ and 0.1％,respectively,without significant difference.Afterwards,the growth of nerve fibers increased in pace and the growth rate is recovered.

The aim of this study is to compare the contents of five types of anthraquinones which mainly includes chrysophanol, physcion, emodin, rhein and physcion and phenolic acids in ten different processed products from Rheum officinale, and to investigate the effect of different initial processing method on the contents of anthraquinones and phenolic acids. Principal component analysis (PCA) was carried out by SPSS software to evaluate the quality of different processed products from Rh. officinale. In conclusion, the contents of anthraquinones in different processed products from Rh. officinale assume the certain regularity. Whether fresh-cut Rheum officinale Bail and how to dry it are derectly effect the contents of anthraquinones and phenolic compounds. The content of anthraquinones in rheum officinale of drying is obviously higher than smudging, and was more abundant in branch root than tap roots. Rh. officinale of drying which growed in Fengjie gained the highest score in PCA. Meanwhile, the procedure of wetting also help to increase the content of anthraquinones and decrease the content of phenolic acids.
Anthraquinones ; chemistry ; Chemistry, Pharmaceutical ; methods ; Drugs, Chinese Herbal ; chemistry ; Hydroxybenzoates ; chemistry ; Rheum ; chemistry

OBJECTIVE:To compare the quality control index between Jinzhen Suanzao teabag and decoction,and explore the alternative supplement of teabag to decoction. METHODS:The test sample solutions of Jinzhen Suanzao teabag and decoction were prepared by hot-maceration method and water-decoction method. The contents of water soluble extract and total flavonoids were de-termined and compared between 2 kinds of preparation. The leaching rates of teabag were investigated at different soak time(0,5, 10,15,20,25,30 min) to optimize soaking time. RESULTS:The average content of water soluble extract were 50.56% and 44.45%(P<0.05) respectively for the teabag and decoction. The total flavonoids content were 0.64 mg/g and 0.69 mg/g (P<0.05). The dissolution amount of teabag were increasing and leaching rate increased within first 20 min,and reached balance gradu-ally 25 min later. CONCLUSIONS:According to the convenience of use and results of each index,the difference in quality control index is not great between 2 kinds of preparation. Teabag can be as the supplement of decoction.

Objectives To explore a better inhibitory effect of concentration and time of Celastrol on migration of HepG2 cells. Methods HepG2 cells were planted and cultured in 6-well plates. When the adherent cell density reached 70%-80%, cell migration was manufactured by scratch experiment model. Then, cell morphology and cell migration were observed under microscope with different concentrations of Celastrol 5, 1, 0.5, 0.1, 0.01μmol/L and DMSO at 0, 6, 12, 24 h. They were pictured and rates of cell migration and inhibition ratios of all groups were calculated. Results Celastrol inhibited HepG2 cell migration, and its inhibitory effect on the migration velocity was concentration-dependent in a certain range. The higher the concentration of Celastrol, the stronger effect is. Celastrol of the same concentration at different times had different inhibitory effect on cell migrationof HepG2 cells (P<0.05). Celastrol of different concentrations at the same time had different inhibitory effects on cell migration of HepG2 cells (P<0.05);Celastrol of high concentration cause dsevere changes in the cell morphology. Conclusion Celastrol of high concentration causes changes in the cell morphology and cell apoptosis of HepG2 cells. Celastrol inhibits HepG2 cell migration, which is dependent on the concentration and action time. The inhibitory effect of Celastrol on HepG2 cell migration is most obvious under final concentration of 5μmol/L at 6 h.

[ ABSTRACT] AIM: To observe the effect of circadian rhythms on wound healing of mouse corneal epithelium. METHODS:The C57BL/6 male mice were used in the study.A part of corneal epithelium (2 mm in diameter) was struck off by a golf-like knife to form a round wound area.The dynamics of epithelial healing in the wound area were ob-served under microscope with fluorescein staining.In addition, with related antibodies and DAPI, the dynamic changes of the neutrophils, platelets and dividing cells were also investigated.RESULTS:The healing rates in LL group (12 h light/12 h light) and DD group (12 h dark/12 h dark) were obviously slower than that in LD group (12 h light/12 h dark), mainly showing delayed re-epithelialization, decreased epithelial cells, increased diameter of blood vessel, and delayed re-cruitment of neutrophils and platelets, but more cell number.CONCLUSION:Disruption of circadian rhythms significant-ly inhibits the wound healing of corneal epithelium, mainly through delaying the inflammation and re-epithelialization, but aggravating the inflammatory responses.

Objective To analyze the assets and liabilities of secondary public hospitals of Wuhan city in Hubei province ,in order to provide references to control the debt scale of such hospitals .Methods Twenty‐one public secondary public hospitals of Wuhan city were analyzed using official statistics on their asset and liabilities from 2012-2014 .Results Such a timeframe witnessed a year‐on‐year growth of both total assets and liabilities of these hospitals ,yet the growth rate of total assets(35.15% ) falls behind that of total liabilities (53.62% )significantly ;significantly slower growth rate of non‐current assets since 2013 ;higher current liabilities than long‐term liabilities on yearly basis ;year‐on‐year grow th of liquidity rate and quick ratio in general ;significant rise of debt‐to‐assets ratio .Conclusions The overall debt scale of such hospitals keeps growing ,a trend deserving actions and correction .

OBJECTIVETo evaluate the effectiveness of a school-based smoking prevention and control intervention program among elementary school students.

METHODSThrough two phase cluster sampling, 566 pupils in grade 4 and grade 5 of two schools were assigned to intervention group and control group. One year comprehensive smoking intervention was conducted in the intervention group. The assessment was carried out through three questionnaires: pre- and post-intervention, 6-month after intervention.

RESULTSAfter one year intervention, pupils in the intervention group significantly improved their knowledge and attitudes related to tobacco use. The rate of attempting smoking decreased form 7.8% to 2.6% and the rate of passive smoking from 53.6% to 41.8%. The difference between the intervention and control groups was statistically significant. However, several index started to decline at 6-month follow up survey.

CONCLUSIONThe result demonstrated the effectiveness and feasibility of tobacco control in elementary school and the positive effect must be developed.

Case-Control Studies ; Child ; China ; epidemiology ; Female ; Health Knowledge, Attitudes, Practice ; Humans ; Male ; Schools ; statistics & numerical data ; Smoking ; epidemiology ; Smoking Prevention ; Surveys and Questionnaires ; Tobacco Smoke Pollution ; statistics & numerical data

OBJECTIVETo explore the effects of survivin antisense RNA and HSP70 double gene transfection on breast cancer cell line MCF-7.

METHODSMCF-7 cells was transfected with the double-gene vector pIRES2-EGFP-survivin antisense RNA/HSP70 via liposome. After a 72-h transfection, the cells were collected for observation under inverted fluorescent microscope. The changes of survivin mRNA and HSP70 protein expressions in the cells were detected with real-time PCR and Western-blot before and after the cell transfection, and the apoptotic rate of the transfected MCF-7 cells was detected by flow cytometry analysis with Annexin-V-cy5/7AAD double staining.

RESULTSGreen fluorescence was detected in MCF-7 cells transfected with the double-gene expression vector and the empty vector under inverted fluorescent microscope. The expression level of survivin mRNA in the cells was reduced effectively after the transfection with the double-gene expression vector, which also induced obvious cell apoptosis and enhanced the expression level of HSP70 protein as compared with those in MCF-7 cells transfected with the empty vector and the untransfected MCF-7 cells.

CONCLUSIONSurvivin antisense RNA can interfere with the expression of endogenous survivin and induce apoptosis of MCF-7 cells. HSP70 can increase the expression of HSP70 protein in MCF-7 cells.

Apoptosis ; drug effects ; Female ; HSP70 Heat-Shock Proteins ; genetics ; pharmacology ; Humans ; Inhibitor of Apoptosis Proteins ; genetics ; pharmacology ; MCF-7 Cells ; RNA, Antisense ; genetics ; pharmacology ; RNA, Messenger ; genetics ; Transfection

A water-soluble polysaccharide fraction from root of Potentilla anserine was obtained. Gas chromatogram, FT-IR, physical and chemical characteristics of the Potentilla anserine polysaccharide fraction (PAPF) were analyzed. The protective effects of PAPF against the H2O2 induced process of apoptosis of murine splenic lymphocytes were investigated in vitro. Morphological assessment of apoptosis was performed with light microscope and laser scanning confocal microscope. DNA fragmentation was visualized by agarose gel electrophoresis. The amount of apoptotic cells was measured by flow cytometry. The results showed that PAPF is composed of rhamnose, arabinose glucose and galactose. H2O2 (200 micromol x L(-1)) induced apoptosis of murine splenic lymphocytes with the cell volume reduced, cytoplasm and nuclear shrunk and DNA stained non-uniformly. Condensed chromatin and formation of apoptotic body were observed in the apoptotic cells. Apoptotic bodies in the cells treated with PAPF and H2O2 were less than those in H2O2 treatment alone. DNA fragmentation assay showed that PAPF (50, 100, 200, and 400 microg x mL(-1)) obviously reduced H2O2-induced ladder bands. Flow cytometry analysis showed that H2O2 increased the populations of apoptotic sub-G1 cells from 5.60% (control) to 45.40%, and PAPF decreased H2O2-induced apoptosis to 37.80%, 22.70%, 17.70%, and 8.50%, respectively. In conclusion, PAPF reduced H2O2-induced oxidative damage in a dose dependent manner.
Animals ; Apoptosis ; drug effects ; Cells, Cultured ; DNA Fragmentation ; Flow Cytometry ; Hydrogen Peroxide ; pharmacology ; Lymphocyte Count ; Lymphocytes ; cytology ; drug effects ; Mice ; Polysaccharides ; pharmacology ; Potentilla ; Spleen ; cytology

Objective To explore the influence of 30 day feeding test of health food containing caffeine on the physiological state of SD rat.Methods SD rats were randomly divided into five groups according weight :three health food containing caffeine(1.64,3.28,6.57 g/kg),one health food containing reduced caffeine (6.48 g/kg)and the control group. Haematological and biochemical parameters were measured at the end of experiment,and main organ tissue analysis was also performed.Results Weight at the end of 4 week,weight after absolute diet,total weight increased,and total food consumption and food consumption at the end of 4 week in 6.57 g/kg groups of male rats were decreased compared with the control group (P <0.05 ).Conclusion In comparison with 6.48 g/kg group,all those changes may be caused by overhigh caffeine in 6.48 g/kg group.