Objective To study the neuroprotective effect of erythropoietin(EPO) on neonatal rats model with hypoxia-ischemia encephalopathy(HIE).Methods HIE was induced in rats on 7th day of postnatal age by ligation of right common carotid artery,followed by 2 h of hypoxia(80 mL/L O2).The subjects were divided into sham-operated group,control group and EPO group.EPO 4 000 U/(kg?day) was injected daily from day 2 pre-surgery for 9 to 16 days and PBS was injected in the control group.The neuroprotective effect of EPO on HIE model was detected by brain weight,the difference in weights between the ipsilateral(right) and contralateral(left) brain and the function test.In vitro study,the neural progenitor cell line C17.2 under gone apoptosis following an ischemia-like metabolic inhibition.The effect of EPO on the cell line ischemia modle 17.2 was evaluated by detecting Annexin V with flow cytometry.Results The signi-ficant and sustained brain injury in the hypoxia-ischemia and vehicle-treated group was observed and measured by reduction in relative weights of ipsilateral to contralateral and compromised sensorimotor functions in response to postural reflex test,compared with those of sham-operated animals(Pa

OBJECTIVETO establish a RP-HPLC method for the determination of calycosin-7-O-beta-D-glucopyranoside in Radix Astragali, and to analyse the calycosin-7-O-beta-D-glucopyranoside content of ten samples of Radix Astragali, collected from different regions.

METHODA Polaris C18(250 mm x 4.6 mm, 5 microns) column was used and a mixture of methanol-water (30:70) was used as the mobile phase at a flow rate of 1.0 mL.min-1. The column temperature was 25 degrees C and the UV detection wavelength was 254 nm.

RESULTThe calibration curve was in good linearity over the range of 0.0106-2.12 micrograms with the regression equation Y = 3035. 97 X - 14.85(r = 0.9999). The average recovery was 95.8% (n = 5, RSD = 1.3%).

CONCLUSIONThe method is simple, quick, sensitive and reproducible. In all of the samples, the calycosin-7-O-beta-D-glucopyranoside contents differ markedly.

Astragalus membranaceus ; chemistry ; classification ; China ; Chromatography, High Pressure Liquid ; Ecosystem ; Glucosides ; analysis ; Isoflavones ; analysis ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Quality Control ; Species Specificity

This study was aimed to prepare the spraying agent of prescriptions of Miao nationality herb and investigate the effect of Miao nationality herbs spray for serum SOD, MDA, and expression of Fas and Caspase-3 mRNA in lung tissues of silica-treated rats. The healthy SD rats were divided into 5 groups. Silica dust suspension was used in the model establishment of 4 groups. After the model was successfully established, 3 groups were randomly selected and given glucocorticoids atomization inhalation, Miao nationality herbs spray, Miao nationality herbs spray combined with intragastric administration of herbal medicine, respectively. After 40-day treatment, water-solubletetrazolium salt (WST-1) was used in the detection of serum superoxide dismutase (SOD). Thiobarbituric acid (TBA) was used in the detection of malondialdehyde (MDA). The mRNA expression variance of the Fas and Caspase-3 were detected by RT-PCR. The results showed that compared with the silica dust suspension group, the SOD activity of serum in the Miao nationality herbs spray group was significantly increased (P< 0.05). MDA content and the mRNA of Fas and Caspase-3 were significantly lower in the Miao nationality herbs spray group (P< 0.05). It was concluded that Miao nationality herbs spray group was able to increase the SOD activity of serum, decrease MDA content, and obviously decrease the expression of Fas and Caspase-3 of lung tissues among silica dust suspension rats.

Objective:To explore the clinical efficacy,safety and tolerability of nasogastric enteral nutrition (NGEN) on moderate-severe acute pancreatitis (M-SAP).Methods:Sixty patients diagnosed as M-SAP and admitted admitted to Department of Gastroenterology,Affiliated Hospital of Southwest Medical University from November 2013 to August 2014 were randomized to receive enteral nutrition (EN) via a nasogasteic tube (n =30) or a nasojejunal tube (n=30).The intestinal permeability (endotoxin,D-lactic acid),inflammation index (CRP、IL-6、TNF-α),nutrition status,nutrition-related adverse effects (reflux aspiration,diarrhea and abdominal pain worse situation),condition assessment (APACHE Ⅱ and MCTSI),and prognosis (infection complications,mortality,length of hospital stay,hospital expenses)were collected and compared.Results:There was no statistically significant difference in the two groups as for intestinal permeability,inflammation index,evaluation of nutritional status,overall condition assessment,and prognosis (P ＞ 0.05).On the reflux aspiration,the rate of nasogastric tube enteral nutrition group (NG group) was 20％,nasojejunal enteral nutrition group (NJ group) was 3.3 ％,the difference was statistically significant (P ＜ 0.05).Conclusion:Either NGEN or NJEN has the similar effects on disease evolution,nutrition condition,intestinal permeability and prognosis of M-SAP.NGEN is simple and worthy of clinical attention.The nasogastric may more easily lead to reflux aspiration than the nasojejunal route.It requires more randomized controlled study to investigate its safety on SAP.

Objective To investigate the nucleotide-binding oligomerization domain-2(NOD-2)gene expression in deep caries and the effects of NOD-2 agonist muramyl dipeptide(MDP)on the differentiation of human dental pulp cells(hDPC). Methods NOD-2 gene level in deep caries and healthy pulp tissue was determined by real-time quantitative polymerase chain reaction(realtime-PCR). Realtime-PCR,Western blotting and immunofluorescence were performed to evaluate NOD-2 gene and protein expression. Dentin sialoprotein(DSP)protein level was assessed when hDPC were challenged by different concentrations of MDP for 24 hours, and sialophosphoprotein(DSPP), osteocalcin(OCN)mRNA and osteopontin(OPN)protein level were detected at different time points after incubation with 0.1 mg/L MDP. Results NOD-2 mRNA level was higher in pulp tissue of deep caries(0.2610±0.0824) than that in healthy controls(0.0024±0.0002),P<0.05.The expression of NOD-2 gene and protein increased in a time denpendent manner upon stimulation with MDP. Immunofluorescence confirmed that NOD-2 protein was located in cytoplasm. Moreover, 0.1 mg/L MDP augmented DSP protein level. DSPP and OCN mRNA were elevated with time and reached the peak at 12 h and down-regulated. OPN protein level also increased with time. Conclusions Dental pulp NOD-2 expression are up-regulated in pulp tissue of deep caries. MDP may be related to the differentiation of hDPC.

MISA (MicroSAtelite) software was employed to screen SSRs in 68 787 contigs of Swertia mussotii transcriptome sequences. 5 610 SSRs were distributed in 5 099 contigs which accounted for 7.41% of 68 787 contigs. There are 220 kinds of SSR motifs existing in S. mussotii transcriptome. On average, SSRs occurred every 12.60 kb in length. In the SSRs, the tri-nucleotide repeat motif was the most abundant (45.99%), followed by the di-nucleotide (41.62%). AT/TA and AAT/TTA were the main types of motif in di-, tri-nucleotide repeats. The repeat numbers of SSRs which from S. mussotii transcriptome SSRs were mainly from 5 to 10 and motif length of them mostly ranged from 12 bp to 30 bp. A total of 30 651 contigs were annotated, and only 1 447 SSRs were occurred in protein-coding regions. In the six repeat motifs, tri-nucleotide repeats were the most abundant in coding regions (928). There are abundant SSRs in S. mussotii transcriptome with high frequency and various types, indicating their usefulness in theory. This research may lay the foundation for designing the targeted SSR primers and developing SSR molecular markers by mining the information of SSRs loci in S. mussotii transcriptome sequences data.
Data Mining ; Medicine, Tibetan Traditional ; Microsatellite Repeats ; Plants, Medicinal ; genetics ; Swertia ; genetics ; Transcriptome

OBJECTIVETo examine the expression of FLICE-inhibitory protein (FLIP) in juvenile idiopathic arthritis (JIA) and analyze its correlation with synovial inflammation.

METHODSThe expression of FLIP was assessed in 11 JIA and 3 normal synovial tissue samples by reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry. The cell types expressing FLIP were further characterized, and the correlation of FLIP expression with the degree of synovial inflammation, as well as the activity of caspase 8 was then analyzed.

RESULTSRT-PCR revealed the expression of FLIP mRNA in all 11 JIA samples, but not in 3 normal synovial tissues. In JIA, FLIP expression could be found in both the lining and sublining layers, mainly in the macrophage-like cells. Moreover, the expression of FLIP in JIA synovial tissues was positively correlated with the degree of synovial inflammation (r = 0.563, P < 0.05).

CONCLUSIONThe expression of antiapoptotic FLIP in JIA synovial tissue and its correlation to accumulation of inflammatory cells in synovial tissue suggests that FLIP potentially extends the lifespan of synovial cells and thus contributes to the progression of joint destruction.

Adolescent ; Arthritis, Juvenile ; metabolism ; pathology ; CASP8 and FADD-Like Apoptosis Regulating Protein ; genetics ; metabolism ; Caspase 8 ; metabolism ; Child ; Female ; Humans ; Inflammation ; metabolism ; pathology ; Male ; Protein Isoforms ; genetics ; metabolism ; Synovial Membrane ; cytology ; metabolism ; pathology

OBJECTIVETo explore the expression of vascular endothelial growth factor (VEGF) and its receptors (flt-1 and flk-1) in the retina of retinopathy of prematurity (ROP), and its relation to the alteration of retinal blood vessels.

METHODSEighty-six newborn Sprague-Dawley rats were randomly divided into hyperoxia and air groups, then each group was further divided into 1, 3, 7 and 14 days subgroups. The rats in hyperoxia group inhaled 75% oxygen and ROP model was thus set up. These animals were sacrificed respectively after 1, 3, 7 and 14 days, then the retinal endothelial cells were marked by CD34 to observe the change of retinal blood vessels. The expression of VEGF, flt-1 and flk-1 in the retina was measured by immunohistochemistry.

RESULTSThe retinal capillary density index (RCDI) in control group increased as days went on (F = 21.589, P < 0.01, but it was the least on the 7th day in hyperoxia group, after the rats had been returned to air for 7 days, RCDI increased significantly (F = 67.885, P < 0.01); In the control group, the expression of VEGF and flk-1 was the strongest in the retina on the 7th day, the result had significant difference as compared with the 1st and 14th day (P < 0.05). The expression of VEGF and flk-1 on the 7th day in hyperoxia group was weaker than that of control group (P < 0.05). But on the 14th day in hyperoxia group, they were stronger than that of control (P < 0.05). The localization of the expression of flt-1 was changed when blood vessels altered, but there was no significant difference in expression intensity as a whole (P > 0.05).

CONCLUSIONWhen the premature retina was exposed to hyperoxia, the expression of VEGF and flk-1 was reduced, and retinal blood vessels were also decreased; but the expression of VEGF and flk-1 was stronger in retina when premature rats were exposed to relative hypoxia, and the retinal blood vessels also increased significantly. It is concluded that VEGF and flk-1 may play important roles in the development of retinal blood vessels and its change in ROP. However, flt-1 has less effect compared with flk-1.

Animals ; Animals, Newborn ; Disease Models, Animal ; Female ; Hypoxia ; Immunohistochemistry ; Male ; Rats ; Rats, Sprague-Dawley ; Receptors, Vascular Endothelial Growth Factor ; analysis ; Retina ; chemistry ; pathology ; Retinal Diseases ; metabolism ; pathology ; Vascular Endothelial Growth Factor A ; analysis

BACKGROUNDTo investigate the frequency of circulating HBV specific T helper cell and evaluate its association with serum levels of HBV DNA before and during lamivudine treatment in patients with chronic hepatitis B.

METHODSThe frequency of circulating HBV specific T helper cells in response to HBcAg in 25 chronic HBV-infected patients was determined by Elispot assay; serum HBV DNA was quantitated by real-time PCR.

RESULTSThe frequency of HBV specific T helper cell before antiviral treatment (47.30 +/- 25.50 SFCs /1 x 10(6) PBMC) was significantly higher than that at the third month of therapy (23.10 +/- 18.45 SFCs /1 x 10(6) PBMC, P < 0.05). All 8 patients observed dynamically had decreased frequency of HBV specific T helper cell at the third month of therapy; six patients with serum HBV DNA level reduced had higher frequency of HBV specific T helper cell before treatment than 2 patients without serum HBV DNA level decrease.

CONCLUSIONHBV specific T helper cell response at the time of hepatitis flare in chronic hepatitis B patients was significantly augmented compared to that at the time of catabasis.

Adult ; Antiviral Agents ; therapeutic use ; DNA, Viral ; blood ; genetics ; Enzyme-Linked Immunosorbent Assay ; methods ; Female ; Hepatitis B Core Antigens ; immunology ; Hepatitis B virus ; drug effects ; genetics ; immunology ; Hepatitis B, Chronic ; blood ; drug therapy ; virology ; Humans ; Lamivudine ; therapeutic use ; Male ; T-Lymphocytes, Helper-Inducer ; cytology ; drug effects ; immunology

The purpose of the present study was to explore the seizure-induced changes in Bad (Bcl-2-associated death protein), 14-3-3, phosphoBad, Bcl-2 and Bcl-XL expression in the rat model of focal limbic seizure. Unilateral intra-amygdaloid injection of kainic acid (KA) was made to induce seizure. Electroencephalogram (EEG) and regional cerebral flow (r-CBF) were monitored continuously. Diazepam (30 mg/kg) was administered to terminate the seizure. The apoptotic and surviving neurons in the hippocampus were observed by terminal deoxynucleotidyl transferrase-mediated dUTP nick end labeling (TUNEL) and cresyl violet staining, the expression of Bad, 14-3-3, phosphoBad, Bcl-2 and Bcl-XL were detected with immunofluorescence, Western blot and immunoprecipitation. The results showed that TUNEL-positive neurons appeared at 8 h and reached maximum at 24 h following seizure cessation within the ipsilateral CA3 subfield of the hippocampus. Seizure induced the dephosphorylation of Bad and the dissociation of Bad from its chaperone protein 14-3-3 and subsequent dimerization of Bad with Bcl-XL. The expression of phosphoBad decreased and Bcl-2 increased. There was little change in r-CBF after the seizure. These results suggest that seizure leads to a dephosphorylation of Bad and an upregulation of Bcl-2. Dephosphorylation of Bad may be injurious while the upregulation of Bcl-2 may be protective to the brain damage induced by seizures, but not related with r-CBF.
Amygdala ; physiology ; Animals ; Epilepsies, Partial ; chemically induced ; metabolism ; Hippocampus ; metabolism ; Kainic Acid ; Male ; Microinjections ; Phosphorylation ; Proto-Oncogene Proteins c-bcl-2 ; biosynthesis ; genetics ; Rats ; Up-Regulation ; bcl-Associated Death Protein ; metabolism