Background:Inflammation plays an important role in intestinal ischemia-reperfusion injury(IRI),and erythropoietin (EPO)has been reported to have anti-inflammatory activity. Aims:To explore the protective effect of EPO on intestinal IRI and its potential mechanism. Methods:Thirty-two healthy male Sprague-Dawley rats were randomly divided into four groups:sham operation group(sham group),IRI group,EPO group and 740Y-P group. Rats in IRI,EPO and 740Y-P groups were injected intraperitoneally with 0. 9% NaCl,EPO and EPO + 740Y-P,respectively,one hour before the establishment of intestinal IRI model by superior mesenteric artery clamping(45 min)-reperfusion. All rats were sacrificed one hour after reperfusion. Histopathological changes of small intestine were observed;expression of proteins in PI3K/ Akt and NF-κB signaling pathways was measured by Western blotting;expression and secretion of inflammatory cytokines, including interleukin-8(IL-8),tumor necrosis factor-α(TNF-α)and monocyte chemoattractant protein-1(MCP-1)were examined by real-time PCR and ELISA. Results:Compared with sham group,the damage score of small intestine,protein expressions of PI3K,p-Akt and p-NF-κB p65,as well as mRNA expressions and serum levels of IL-8,TNF-α and MCP-1 in IRI group were significantly increased(P < 0. 05). EPO pretreatment could ameliorate the histopathological changes of small intestine in IRI model rats,inhibit PI3K/ Akt and NF-κB signaling activation and down-regulate expression and secretion of inflammatory cytokines. When 740Y-P,a PI3K agonist,was used combinedly,the effect exerted by EPO was diminished. Conclusions:EPO pretreatment can protect against intestinal IRI by inhibiting the activation of PI3K/ Akt/NF-κB signaling and the subsequent inflammatory response.

Objective To explore the relationships between plasma vasoactive intestinal peptide(VIP),superoxide dismutase(SOD),malondialdehyde(MDA) and hypoxic-ischemic brain damage(HIBD) in neonates with asphyxia.Methods Sixty-eight full-term neonates hospitalized with asphyxia were enrolled in this study (simple asphyxia group 15 cases,mild HIE group 17 cases,moderate HIE group 22 cases and severe HIE group 14 cases) according to the diagnostic criteria of neonatal hypoxic- ischemic encephalopathy(HIE),and 20 cases in control group.Plasma VIP,SOD and MDA were detected by radio-immuhoassay,thiobarbatic acid colorimety and xanthine oxidase at d1 and d7 after born in every group.Results 1.There were significant difference in the plasma VIP,SOD and MDA among every group(Pa

Objective:To investigate the radiosensitizing effect of three flavonoids on ovarian cancer SKOV3 cells under hypoxia. Methods:The SKOV3 cells were divided into normoxic group and hypoxic group. The hypoxic SKOV3 cellular model in vitro was es-tablished and tested by measuring the expression profile of HIF-1αprotein in SKOV3 cells. Colony-forming assay was used to detect the radiosensitivity of normoxic and hypoxic SKOV3 cells. The cytotoxicity and radiosensitizing effects of flavonoids were evaluated on the basis of cell death and MTT assay. Results:The Western blot results showed that the gray intensity ratio of HIF-1α/β-Actin in hypoxia group was significantly higher than that in normoxia group (1. 068>0. 117). Radiosensitivity of hypoxic SKOV3 cells in hypoxia group was significantly lower than that in normoxia group. The survival rate of SKOV3 cells was decreased with the increase of concentration. When the concentration was increased, D0 and Dqin chrysin group and quercetin group were significantly decreased (P<0. 01), and SER was increased (P<0. 05). SER showed no significant difference in breviscapine group (P>0. 05). The overall radiobiological parameters of hypoxia group were higher than those of normoxia group. Conclusion: Hypoxia can induce the expression of HIF-1α in SKOV3 cells, which results in the decrease of radiosensitivity. Chrysin and quercetin can enhance the radiosensitivity of SKOV3 cells, and the enhancement is significant under hypoxia, while breviscapine is without such effect. The radiosensitizing effect may be achieved by the level decrease of HIF-1α in SKOV3 cells and inhibition of DNA damage repair.

Objective To estimate the present salt iodine content and iodine nutrition need of high risk population of iodine deficiency disorder in Meixian County. Methods Each primary school was selected from urban and rural areas(Xiyang Town, 20 kilometers away from Meixian County), the goiter rate of 8 to 10 year-old students was examined and urinary iodine and household salt iodine was sampled. Twenty to 40 year-old women of childbearing age nearby schools around the urban and villages around Xiyang Town were selected to collect their urine and salt samples. At urban hospitals and rural health centers, 0 to 2 year-old infant urine samples were collected, Thyroid gland was palpated and urinary iodine was determined by iodine in urine by As3+-Ce4+catalytic spectrophotometry, salt iodine was determined by direct titration. Results The goiter rates of 8 to 10 year-old students were 1.5 % (3/200), 1.0% (1/100) for the urban area and 2.0% (2/100) for rural area. Median of urinary iodine in 8 to 10 year-old students, infants, women of childbearing age averaged at 237.1 μg/L and 280.1, 234.7,187.6 μg/L respectively, with each being 287.4,245.0,205.5 μg/L in urban area and 278.9,228.5,176.4 μg/L in rural area. Women of childbearing age had a higher percentage of urinary iodine < 50.0 μg/L than students,students had a higher percentage than infants, each being 7.5%(15/200), 4.5%(9/200), 4.0%(4/100). The ration of urinary iodine > 300.0 μg/L was more in infants than in students, that in students was more than that in women of childbearing age, each being 33.0% (33/100), 30.0% (60/200),22.5% (45/200). The median of salt iodine was 27.2 mg/kg. The coverage of iodized salt was 100.0%(400/400). Ninty-seven percent(194/200) and 96.0% (192/200) of qualified iodized salt were consumed in urban area and in rural area. Conclusions The amount of iodine added to salt meets the requirement in the 3 kinds population risk of iodine deficiency disorder. But a higher iodine status has been found out in students and infants. It is reasonable to decrease the present salt iodine content.

Objective To prepare triamcinolone acetonide acetate(TAA)thermosensitive hydrogel for intra-articular injection,and to investigate its release in vitro. Methods TAA suspending thermosensitive hydrogel was prepared by using poly lactic-co-glycolic acid(PLGA)-polyethylene glycol(PEG)-PLGA as gel matrices,xanthan gum as suspending agent and NaCl as flocculant. It was evaluated preliminarily by determining its contents and its in vitro release. Results The best compositions for preparation of TAA suspending thermosensitive hydrogel were 25% PLGA-PEG-PLGA,0.05% xanthan gum, 0.9% NaCl and 4 mg·mL-1 TAA.The gelation temperature was 35.3 ℃ .The average recovery was(98.98±0.31)%. By the method of membraneless dissolution,the accumulative drug release was up to 83. 31% at 16 days. The drug release followed Ritger-Peppas methematical model. Conclusion The TAA thermosensitive hydrogel with obviously sustained release is expected to become a new drug delivery system for intra-articular injection.

OBJECTIVE:To optimize the formulation of Methotrexate thermosensitive hydrogels,and to study the in vitro re-lease property of it. METHODS:Taking PLGA-PEG-PLGA as matrix and mannitol as viscosity modifiers,Methotrexate thermosen-sitive hydrogels were prepared. Using the absolute value of the deviation of gelation temperature between 35 ℃,its formulation was optimized by orthogonal test using the concentrations of PLGA-PEG-PLGA,methotrexate and mannitol as factors. The dialysis bag method was used to investigate accumulative release rate of Methotrexate thermosensitive hydrogels and methotrexate solution within 336 h in vitro. RESULTS:The optimal formulation was as PLGA-PEG-PLGA of 20%,methotrexate of 0.10% and mannitol of 0.10%;gelation temperature were 35.1,35.0,35.0℃. The average drug-loading rate was more than 90%. 12 d accumulative re-lease rate was more than 90%(r=3). Methotrexate solution has been substantially completely relased within 240 h. CONCLU-SIONS:Methotrexate thermosensitive hydrogels with sustained-release are prepared successfully,and the preparation technology is simple and practical.

Chemical constituents of Leonurus japonicus were isolated and purified by a combination of various chromatographic techniques including column chromatography over silica gel, Sephadex LH-20, MCI, and Rp C18. Structures of the isolates were determined by spectroscopic analysis as 10 coumarins: bergapten (1), xanthotoxin (2), isopimpinellin (3), isogosferal (4), imperatorin (5), meransin hydrate(6), isomeranzin(7), murrayone(8) , auraptenol(9), and osthol(10). In addition to compound 9, the others were isolated from the genus Leonurus for the first time. In the in vitro assay, compounds 4 and 8 significantly inhibited the abnormal increase of platelet aggregation induced by ADP.
Blood Platelets ; drug effects ; Coumarins ; chemistry ; pharmacology ; Leonurus ; chemistry ; Platelet Aggregation ; drug effects ; Platelet Aggregation Inhibitors ; chemistry ; pharmacology

Objective:To investigate the cytopathic effect of amino acids 86-175 of rotavirus non-structural protein 4 (NSP4 86-175) on rat cardiomyocytes and the possible mechanism. Methods:Rat H9C2 cardiomyocytes were treated with NSP4 86-175 protein. Changes in the growth and morphology of the cells were observed. The activity of LDH in the cell culture medium was detected. Fluo-3AM was used to label intracellular free calcium ions and the concentrations of calcium ions in rat cardiomyocytes with and without NSP4 86-175 treatment were detected by confocal laser microscopy. The expression of Bax, Bcl-2, caspase-3, 78 kDa glucose-regulated protein (GRP78), C/EBP homologous protein (CHOP) and caspase-12 at mRNA level was detected by real-time PCR. The expression of caspase-3, caspase-8, caspase-9, GRP78, CHOP and caspase-12 at protein level was detected by Western blot. Results:Normal cardiomyocytes showed a typical myoblast-like morphology, presenting as spindle-shaped cells with clear boundaries. Obvious cytopathic effect, vacuolar degeneration, shriveled and rounded cells, and cell fragmentation were observed after the treatment with purified NSP4 86-175 protein. The activity of LDH in cell culture medium was enhanced by NSP4 86-175 protein. NSP4 86-175 protein also enhanced the fluorescence of the calcium ions in rat cardiomyocytes, promoted cell apoptosis, up-regulated the expression of apoptotic factors including caspase-3, caspase-8, caspase-9 and Bax-2, and increased the expression of classical markers of endoplasmic reticulum stress such as GRP78, CHOP and caspase-12. Conclusions:NSP4 86-175 had a cytopathic effect on rat cardiomyocytes, which might be related to the induced intracellular calcium overload, endoplasmic reticulum stress, apoptosis and necrosis. These results might be used as theoretical reference for further study on rotavirus infection and myocardial injury.

OBJECTIVETo study the inhibition effect of HIF-1α specific siRNA expression vector pSUPERH1-siHIF-1α on retinal neovascularization in a mouse model of retinopathy of prematurity (ROP).

METHODSThe mouse model of ROP was prepared by the method Smith described. Forty-eight ROP mice were randomly divided into two groups: an experimental group that was intravitreously injected with pSUPERH1-siHIF-1α and a control group that was injected with pSUPER retro vector. The levels of HIF-1α and vascular endothelia growth factor (VEGF) in the retina were examined by Western blot. The retinal neovascularization was evaluated by angiography using FITC Dextran and quantitated histologically.

RESULTSThe levels of HIF-1α and VEGF in the retina in the experimental group were reduced 90% and 65% respectively compared with those in the control group. Meanwhile, the number of retinal neovascular endothelial nucleus outbreaking the inner limit membrane in the experimental group was significantly reduced compared with that in the control group.

CONCLUSIONSThe development of retinal neovascularization of ROP can be markedly inhibited by RNA interference targeting HIF-1α.

Animals ; Disease Models, Animal ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; analysis ; antagonists & inhibitors ; genetics ; Infant, Newborn ; Mice ; Mice, Inbred C57BL ; RNA, Small Interfering ; genetics ; Retinal Neovascularization ; prevention & control ; Retinopathy of Prematurity ; therapy ; Vascular Endothelial Growth Factor A ; analysis

SIP24/24p3 is a secreted murine acute phase protein which has been speculated to play an anti-inflammatory role in vivo. Recently SIP24/24p3 has been found to be able to specifically induce apoptosis in leukocytes. By using (35)S metabolic labeling method, we studied the regulation of SIP24/24p3 by glucocorticoid and pro-inflammatory cytokines IL-6 and TNF-alpha in cultured Balb/c 3T3 and BNL cells. The following results were observed: (1) dexamethasone induced the expression of SIP24/24p3 in both Balb/c 3T3 and BNL cells, the induction was more significant in BNL cells; (2) dexamethasone and IL-6 synergistically induced the expression of SIP24/24p3 in both Balb/c 3T3 and BNL cells; (3) in Balb/c 3T3 cells dexamethasone and TNF-alpha acted synergistically to induce the expression of SIP24/24p3, whereas in BNL cells dexamethasone and TNF-alpha induced the expression of SIP24/24p3 in an additive manner; (4) dexamethasone and IL-6/TNF-alpha acted synergistically in Balb/c 3T3 cells and additively in BNL cells to induce the expression of SIP24/24p3. The inducibility of SIP24/24p3 by multiple factors will help to explain its highly specific expression in vivo. The difference in the expression patterns of SIP24/24p3 in different cell types is also suggestive to its expression and regulation in hepatic and extrahepatic tissues. Finally, the fact that SIP24/24p3 protein can be induced by both pro-inflammatory as well as anti-inflammatory factors is indicative of the important role of SIP24/24p3 in the entire acute phase response process.
Acute-Phase Proteins ; biosynthesis ; genetics ; Animals ; BALB 3T3 Cells ; Carrier Proteins ; biosynthesis ; genetics ; Cytokines ; pharmacology ; Dexamethasone ; pharmacology ; Drug Synergism ; Gene Expression Regulation ; Interleukin-6 ; pharmacology ; Lipocalin-2 ; Lipocalins ; Mice ; Mice, Inbred BALB C ; Oncogene Proteins ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Tumor Necrosis Factor-alpha ; pharmacology