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
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chemistry
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Candida albicans
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drug effects
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Cell Line, Tumor
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Clausena
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chemistry
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HeLa Cells
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Humans
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Magnetic Resonance Spectroscopy
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Molecular Structure
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Plant Leaves
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chemistry
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Plant Roots
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chemistry
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Plant Stems
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chemistry
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Staphylococcus aureus
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drug effects
7.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
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chemistry
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Magnetic Resonance Spectroscopy
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Molecular Structure
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Plant Components, Aerial
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chemistry
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Plants, Medicinal
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chemistry
8.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
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Alpinia
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chemistry
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Chromatography, High Pressure Liquid
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Fruit
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chemistry
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Phytochemicals
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chemistry
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isolation & purification
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Sesquiterpenes
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chemistry
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isolation & purification
9.Two-step Tandem Chromatography Purification of Anti-human CD80 Monoclonal Antibody 4E5 from Mouse Ascites
Hong-Bing MA ; Yu-Hua QIU ; Ran TAO ; Wen-Xiang LI ; Ying XU ; Xue-Guang ZHANG ;
China Biotechnology 2006;0(08):-
A two-step method for the purification of blocking-type anti-human CD80 monoclonal antibody 4E5 from mouse ascites was developed using anion exchange and gel filtration in combination. The ascites was first purified by anion exchange after centrifugation and filtration. The experimental parameters of sample loading and elution were optimized. The optimized loading condition was pH 8.0,50 mmol/L Tris-HCl and satisfactory results were obtained using a 0~0.5mol/L NaCl step elution. The fraction containing the protein of interest was directly loaded on gel filtration column and eluted using a 20 mmol/L phosphate buffer at pH 7.2. The purity of the obtained monoclonal antibody was up to 95% with a recovery of 61%. The purity of mAb could efficiently inhibit the growth of Daudi cells. The amplification of the method was also studied using a Bio-Scale Q5 column and the result was satisfied.
10.Effects on osteoblast and adipocyte differentiation of bone marrow mesenchymal stimulated by pulsed electromagnetic fields
Yong YANG ; Hua WU ; Dong-Ming ZHAO ; Wen-Chun ZHAO ; Run-Guang LI ;
Chinese Journal of Physical Medicine and Rehabilitation 2003;0(08):-
Objective To study the effects of pulsed electromagnetic fields(PEMFs)of 15Hz and lmT on osteoblast differentiation of bone marrow mesenchymal stem cells.Methods The bone marrow mesenchymal stem cells were isolated and cultured in vitro,the third passage cells were harvested and half of them was exposed to PEMFs,another half exposed to sham PEMFs served as the control.The activity of ALP was measured.And the ex- pression of osteoblast and adipogenesis was measured with the semi-quantitative RT-PCR,adipogenic differentiation was observed with the Oil Red O Staining.Results When compared with the control,PEMFs of 15 Hz and 1 mT significantly increased the activity of ALP(P