3.Model of diabetic keratopathy with high-fat and high-glucose feeding mice
Wen, BO ; Guang-Hua, SUN ; Feng-Xia, SUN ; Wen, CUI
International Eye Science 2015;(4):596-600
AIM: To discuss the establishment of immediate diabetic keratopathy animal model of C57BL/6 mouse induced by ahigh-fat and high-glucose diet.
METHODS: Diabetes mellitus was induced by a high-fat and high-glucose diet in C57BL/6 mouse. 1% rose bengal was stained on the cornea to examine the integrality of the corneal epithelium at 2 ~ 12mo after completion of the model. Corneal epithelial wound healing was observed using a vivo epithelial debridement model which was dyed by sodium fluorescein. Corneal morphology histology was examined by pathological methods.
RESULTS: The high-fat and high-glucose diet C57BL/6 mouse in 2mo had showed general symptoms of diabetes: polydipsia, polyphagia, polyuria, weight loss etc. The model had a steady-state high glucose (≥18mmol/L), also the weight was lower compared with normal control mouse. 1% rose bengal corneal staining had dot coloring at 2mo after completion of the model, the stained area and extent were gradually increased with the extension of the duration of diabetes, almost all the cornea was stained at 12mo after completion of the
model. With the passage of time into a mold, the cornea epithelial healing time become longer: 2mo was about 40h;3mo was about 120h; 4, 6, 12mo was about 144h;the coloboma were gradually increased at 12mo after completion of the model, then the area was reduced gradually until complete healing, the time was 96~120h, showed repeating phenomenon.
CONCLUSION: The mouse were induced by high-fat and high-glucose diet can be used as animal models of diabetic keratopathy: the damage of epithelium for corneal and delay healing on epithelium and other symptoms.
4.Effect of Actinidia chinensis planch polysaccharide on the growth and apoptosis, and p-p38 expression in human gastric cancer SGC-7901 cells.
Wen-Ying SONG ; Guan-Hua XU ; Guang-Ji ZHANG
Chinese Journal of Integrated Traditional and Western Medicine 2014;34(3):329-333
OBJECTIVETo investigate the effect of Actinidia chinensis Planch polysaccharide (ACPS) on the growth and apoptosis of human gastric cancer SGC-7901 cells, and to explore the effect of SGC-7901 cells on p-p38 expression.
METHODSThe inhibition rates at different concentrations of ACPS on SGC-7901 cells at 24, 48, and 72 h were detected using CCK-8 method. Apoptosis ratios in SGC-7901 were determined by flow cytometry after 48-h treatment of different concentrations of ACPS. The expression of pro-caspase-9, PARP, and p-p38 in SGC-7901 cells after treated by different concentrations of ACPS was detected using Western blot. The expression of pro-caspase-9, PARP, and p-p38 was detected after SGC-7901 cells were pre-treated by p38 specific inhibitor.
RESULTSCompared with the control group, the optical density of SGC-7901 cells decreased after treated by 1, 2.5, 5, and 10 mg/mL ACPS (P < 0.05). Meanwhile, the longer the acting time, the lower the optic density (P < 0.01). IC50 was 7.43 mg/mL at 24 h; 3.88 mg/mL at 48 h, and 1.32 mg/mL at 72 h respectively. ACPS suppressed the protein expression of pro-caspase-9 (P < 0.01) and up-regulated the expression of PARP (89KD) (both P < 0.01). Further study showed that the protein expression of p-p38 was up-regulated in SGC-7901 cells treated by ACPS of different concentrations at 24 h (P < 0.05). The expression of phosphorylation p38 and the ACPS induced apoptosis of SGC-7901 cells could be inhibited after treated by specific inhibitor for 2 h.
CONCLUSIONSACPS could inhibit the growth of SGC-7901 cells and induce apoptosis. The underlying mechanism of inducing apoptosis was partially due to activating the p38MAPK path and further activating Caspase9 and PARP, finally leading to cell death.
Actinidia ; chemistry ; Apoptosis ; drug effects ; Caspase 9 ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Humans ; Poly (ADP-Ribose) Polymerase-1 ; Poly(ADP-ribose) Polymerases ; metabolism ; Polysaccharides ; pharmacology ; Stomach Neoplasms ; metabolism ; pathology ; p38 Mitogen-Activated Protein Kinases ; metabolism
5.Clinical Significance and Influence on Glucocorticoid Receptor Resulted from Tripterygium Wilfordii Hook F for Children with Nephrotic Type of Henoch-Schonlein Purpura Nephritis
tai-guang, ZHOU ; zheng-hua, DENG ; shan-wen, HUANG
Journal of Applied Clinical Pediatrics 2006;0(17):-
Objective To explore the mechanism of tripterygium wilfordii hook f(TWHF)for Henoch-Schonlein purpura nephritis(HSPN,nephrotic type)in children.Methods All the children with HSPN(nephrotic type)were divided into 3 groups casually.Method of radioactive pairs conjugating analysis was used to determine the numbers of glucocorticoid(GC)receptors(GCRs).Results were statistically analysed respectively.Results GCRs tested in all the patients with HSPN(nephrotic type)before treatment were statistically lower than those in control group of normal children;GCRs in all the patients tested after 1 week treatment with GC were lower than before treatment.Three weeks later,GCRs in patients treated with TWHF increased much higher than them without TWHF.Following up the recurring times of all the patients 3 months to 5 years,they were much more in patients treated without TWHF treatment than in patients treated with it.Conclusions TWHF treatment on HSPN(nephrotic type)is obviously effective.It can not only decrease the dosage of GC,but decrease the recurring times.One of the mechanism may be TWHF can resist the down-regulation GC to GCR,and enhance the effect of GC.
6.Twelve benzene derivatives from Clausena excavata.
Wen-Wen PENG ; Wei-Wu SONG ; Mao-Bo HUANG ; Guang-Zhi ZENG ; Ning-Hua TAN
Acta Pharmaceutica Sinica 2014;49(12):1689-1693
A new phenethanol, (2'R)-4-(2', 3'-dihydroxy-3'-methyl-butanoxy)-phenethanol (1), along with other eleven known benzene derivatives (2-12) were isolated from the roots, stems and leaves of Clausena excavata (Rutaceae). Compounds 3 and 4 are new natural products, and compounds 5-8, 10-12 were isolated from C. excavata for the first time. Their structures were elucidated on the basis of MS, 1D and 2D NMR spectroscopic analyses including HSQC, COSY and HMBC experiments. 1 was tested for its cytotoxicities against A549, HeLa and BGC-823 cancer cell lines, and antimicrobial activities against Candida albicans and Staphylococcus aureus. The results showed that 1 did not exhibit cytotoxic and antimicrobial activities.
Benzene Derivatives
;
chemistry
;
Candida albicans
;
drug effects
;
Cell Line, Tumor
;
Clausena
;
chemistry
;
HeLa Cells
;
Humans
;
Magnetic Resonance Spectroscopy
;
Molecular Structure
;
Plant Leaves
;
chemistry
;
Plant Roots
;
chemistry
;
Plant Stems
;
chemistry
;
Staphylococcus aureus
;
drug effects
7.The biocompatibility of recombinant human type-Ⅲ collagen-based tissue-engineered cornea in rabbit eye
Hui-feng, RONG ; Qing, ZHAO ; Hua, YAN ; Gui-pei, LIU ; Yuan, LIU ; Wen-guang, LIU
Chinese Journal of Experimental Ophthalmology 2011;29(12):1065-1070
Background The tissue-engineered cornea is becoming the hot spot in the ophthalmologic field,while the research of corneal substitute is in the ascendant,because it is more similar to the corneal morpha and easy to survive in vivo.Objective This study was to investigate the biocompatibility of recombinant human type-Ⅲ collagen/poly9 ( 3-( methacryloylamino ) propyl dimethyl ( 3-sulfopropyl ) ammonium hydroxide ) ( PMPDSAH ) interpenetrating polymer network (IPN) (RHC-Ⅲ/PMPDSAH IPN) hydrogel as a tissue-engineered cornea in rabbit eye and its feasibility as the corneal substitute.Methods One hundred and eight rabbits were randomly divided into experimental group( 90 rabbits) and normal control group ( 3 rabbits),and 15 rabbits ( 30 eyes ) used as the donor corneas.RHC-Ⅲ/PMPDSAH IPN,NGF PMPDSAH IPN and corneal grafts were lamellarly transplanted into the right eyes in RHC-Ⅲ/PMPDSAH IPN group,NGF PMPDSAH IPN group and allograft group respectively.The corneal transparency and neovascularization were examined and scored under the slim lamp and compared among three groups using Kraskal-Wallis H test.The corneal epithelization time was observed and compared among these three groups using one way analysis of variance and LSD-t test.The histological examination of corneas was performed at the 3rd day,1st and 2nd week,1 st,3rd and 6th month after the surgery.The immunohistochemistry was used to detect the expression of K3 in cornea at the 6th month.Results The grafts were well attached in RHC-Ⅲ/PMPDSAH IPN group,NGF PMPDSAH IPN group and allograft group,and no rejection reaction was found throughout 6-month following up.Compared with normal control group,no significant differences were found in the scores of corneal opacification and neovescularization in these three groups (x2 =4.34,P =0.23 ;x2 =2.60,P =0.46 ) at the 6th month.NGF PMPDSAH IPN group achieved reepithelialization in (4.97±0.63) days and was obviously shorted than that in RHC-Ⅲ/PMPDSAH IPN group and allograft group ( t =11.97,P =0.00; t =5.80,P =0.00).The re-epithelialization time in RHC-Ⅲ/PMPDSAH IPN was (6.86±0.71) days,and that of allograft group was (5.87±0.43 ) days,showing a significant difference ( t =6.32,P =0.00).Hematoxylin-eosin staining results demonstrated that implanted materials integrated into the host corneal tissue well and support corneal epithelialization.Part of the material degraded at the 2nd week and degraded completely 1 month later.Regular alignment and distribution of collagen fibers were seen in the regenerated cornea and were similar to those of the normal stroma in 6 months.Immunohistochemistry showed the positive expression of keratin-3 in corneal epithelial cells.Conclusions RHC-Ⅲ/PMPDSAH IPN has a good biocompatibility without toxicity to corneal tissue.Furthermore,NGF can promote the corneal wound-healing and re-epithelialization.The material can be used as safe and reliable corneal substitute after improving the mechanical strength.
8.New dicyclopeptides from Dianthus chinensis.
Jing HAN ; Zhe WANG ; Yu-Qing ZHENG ; Guang-Zhi ZENG ; Wen-Jun HE ; Ning-Hua TAN
Acta Pharmaceutica Sinica 2014;49(5):656-660
One new dicyclopeptide cyclo-(L-N-methyl Glu-L-N-methyl Glu) (1), together with one new natural dicyclopeptide cyclo-(L-methyl Glu ester-L-methyl Glu ester) (2), and two known dicyclopeptides cyclo-(L-methyl Glu ester-L-Glu) (3), and cyclo-(L-Glu-L-Glu) (4), were isolated from the aerial parts of Dianthus chinensis L. Their structures were determined by spectroscopic analyses and chemical methods.
Dianthus
;
chemistry
;
Magnetic Resonance Spectroscopy
;
Molecular Structure
;
Plant Components, Aerial
;
chemistry
;
Plants, Medicinal
;
chemistry
9.The compounds from n-butanol fraction of Alpinia oxyphylla.
Bin-Bin XIE ; Lei HOU ; Bao-Lin GUO ; Wen-Hua HUANG ; Jing-Guang YU
Acta Pharmaceutica Sinica 2014;49(11):1569-1573
Nine compounds were isolated from the n-butanol fraction of 95% ethanol extract of the fruit of Alpinia oxyphylla Miq. with a combination of various chromatographic approaches, including MDS resin, silica gel, reverse phase C18 and preparative HPLC. On the basis of spectroscopic data analysis, they were elucidated as (1R, 4R, 10R)-1β, 4α-dihydroxy-11, 12, 13-trinor-5, 6-eudesmen-7-one (1), 1β, 4β-dihydroxy-11, 12, 13-trinor-8, 9-eudesmen-7-one (2), oxyphyllenone A (3), oxyphyllenone B (4), rhamnocitrin (5), staphylionoside D (6), benzyl-1-O-β-D-glucopyranoside (7), 2-O-β-D-glucopyranosyl-(1S)-phenylethylene glycol (8), and (S)-1-phenylethyl-β-D-glucopyranoside (9). Among them, compound 1 is a new sesquiterpene, named as oxyphyllenone C; compounds 8 and 9 are new natural products; compounds 2 and 6 were isolated from the genus Alpinia for the first time, and compound 7 was isolated from A. oxyphylla for the first time.
1-Butanol
;
Alpinia
;
chemistry
;
Chromatography, High Pressure Liquid
;
Fruit
;
chemistry
;
Phytochemicals
;
chemistry
;
isolation & purification
;
Sesquiterpenes
;
chemistry
;
isolation & purification
10.Effects of bone marrow-derived mesenchymal stem cells engraftment on vascular endothelial cell growth factor in lung tissue and plasma at early stage of smoke inhalation injury
Feng ZHU ; Guang-Hua GUO ; Wen CHEN ; Nian-Yun WANG
World Journal of Emergency Medicine 2010;1(3):224-228
BACKGROUND: This study was undertaken to determine the effect of mesenchymal stem cells (MSCs) engraftment on vascular endothelial cell growth factor (VEGF) in lung tissue, plasma and extravascular lung water at early stage of smoke inhalation injury. METHODS: A rabbit smoke inhalation injury model was established using a home-made smoke inhalation injury generator, and rabbits were divided into two groups randomly: a control group (S group, n=32) and a MSCs treatment group (M group,n=32). 10 ml PBS was injected via the ear marginal vein immediately at injury into the S group. Third generation MSCs with a concentration of 1×107/10 ml PBS were injected via the ear marginal vein immediately at injury into the M group. VEGF in peripheral blood and lung tissue were measured at 0 (baseline), 2, 4 and 6 hours after injection respectively and analyzed. The right lungs of rabbits were taken to measure lung water mass fraction. RESULTS: In the lung tissue, VEGF decreased gradually in the S group (P<0.05) and signifi cantly decreased in the M group (P<0.05), but it increased more signifi cantly than the values at the corresponding time points (P<0.05). In peripheral blood, VEGF increased gradually in the S group (P<0.05) and markedly increased in the M group (P<0.05), but it decreased more signifi cantly than the values at corresponding time points (P<0.05). CONCLUSION: MSCs engraftment to smoke inhalation injury could increase VEGF in lung tissue, decrease VEGF in plasma and reduce extravascular lung water, indicating its protective effect on smoke inhalation injury.