Objective To examine the dyllamic changes in excitatory and inhibitory amino acid neurotransmitter release in the spinal cord in a rat model of incisional pain.Methods Twelve healthy adult male SD rats weighing 250-300g were anesthetized with intraperitoneal chloral hydrate 300 mg/kg.A loop microdialysis catheter was implanted into the subarachnoid space via the atlanto-occipital membrane and advanced for 8.5 cm candad until lumbar region.The animals were randomly divided into 2 groups(n=6 each): control group(C) and incisional pain group(I).Incisional pain was produced by the plantar incision in the tight hindpaw under 1.2% isoflurane in group I while group C received only anesthesia with 1.2% isoflurane.The microdialysis samples were collected before incision(To,baseline)at 3 h,1 d,2 d and 3 d after incision(T1-4) for determination of amino acid using HPLC.The pain behavior was assessed and scored (O=no pain,2=severe pain) at the above time paints.Results In group I the aspartate and glutamate concentrations in the microdialysis samples were significantly increased at 3 h after incision(T1) as compared with the baseline value at To and returned to the baseline level at l d(T2);the glyeine and r-amino butyric acid concentrations were signifieantly increased at ld (T2)and returned to the baseline level at 2 d(T3).The cumulated pain scores were significantly increased at 3 h,1 d and 2 d after incision and returned to baseline level at 3 d (T4) in group I.Conclusion The increased release of excitatory amino acid neurotransmitter in the early phase after incision may be involved in hyperalgesia while the increased release of inhibitory amino acid neurotransmitter in the later phase may be involved in the pain relief.

Objective To establish P2 or P0 peptide-induced experimental autoimmune neuritis (EAN)in the Lewis rats and to explore the optimal type and doses of antigen inoculated to induce EAN and the associated cell-mediated immune mechanisms.Methods Lewis rats were classified into EAN and control groups.The EAN rats were immunized by injection into both hind footpads of inoculums containing 100 or 200 ?g of P2_(57-81)peptide or 200?g of P0_(180-199)peptide and Freund's complete adjuvant(FCA),and the control rats were immunized with FCA only.Clinical scores were compared when they were at peak time of paralysis.Lymphocyte proliferation assay,the ratio of CD_4~+ T cells to lymphatic monocytes and percentage of CD_4~+ CD_(25)~+ T cells to CD_4~+ T cells obtained on day 14 post-immunization were examined.Histopathalogical assessment of sciatic nerves was made.Results Peak clinical scores of P2_(57-81)200 ?g group were dramatically higher than those in P2_(57-81)100 ?g group and P0_(180-199)200 ?g group(both P

Objective To evaluate the role of Phosphatidylinositol 3-kinase/protein-serine-threonine kinases (PI3K/Akt) signaling pathway in reduction of hypoxia-reoxygenation (H/R)-induced injury to cardiomyocytes by propofol postconditioning in rats.Methods Primary cardiomyocytes were obtained from neonatal rats aged 1-3 days and cultured in DMEM culture medium.The cells were seeded in 96-well plates (density 1 × 105/ml,200 μl/well) or 6-well plates (density 5 × 105/ml,2 ml/well) and randomly assigned into 4 groups (n =24 each):control group (C group),H/R group,H/R + propofol group (H/R + P group) and H/R + propofol + wortmannin (a specific PI3K inhibitor) group (H/R + P + W group).The cells were routinely cultured for 6 h in group C.The cells were exposed to 2 h hypoxia followed by 4 h reoxygenation.Propofol with the final concentration of 50 μmol/L was added to the culture medium at the end of hypoxia in group H/R + P.Wortmannin (final concentration 100 nmol/L) and propofol (final concentration 50 μmol/L) was added to the culture medium at the end of hypoxia in group H/R + P + W.At the end of reoxygenation,the cell viability was measured by MTT assay,the lactic dehydrogenase (LDH) activity was detected in the culture medium,the cell apoptosis was assessed by flow cytometry,and the expression of phosphorylated Akt (p-Akt),Bcl-2 and Bax in cardiomyocytes were determined by Western blot.The apoptotic rate and Bcl-2/Bax ratio were calculated.Results Compared with C group,the cellviability was significantly decreased,the LDH activity and apoptotic rate were increased,p-Akt and Bax expression was up-regulated and Bcl-2 expression was down-regulated in H/R group (P ＜ 0.05).Compared with H/R group,the cell viability and Bcl-2/Bax ratio were significantly increased,the LDH activity and apoptotic rate were decreased,p-Akt and Bcl-2 expression was up-regulated and Bax expression was down-regulated in H/R + P group (P ＜ 0.05).Compared with H/R + P group,the cell viability and Bcl-2/Bax ratio were significantly decreased,the LDH activity and apoptotic rate were increased and p-Akt and Bel-2 expression was down-regulated in H/R +P + W group (P ＜ 0.05).Conclusion The mechanism by which propfol postconditioning attenuates H/R-induced injury to cardiomyocytes is related to the activation of PI3K/Akt signaling pathway.

Antibodies ; immunology ; Antibody Formation ; Complement C4b ; immunology ; pharmacology ; Diagnosis ; Graft Rejection ; diagnosis ; immunology ; Humans ; Kidney Transplantation ; immunology ; Peptide Fragments ; immunology ; pharmacology ; Transplantation, Homologous ; immunology

Objective To investigate the correlation of killer immunoglobulin-like receptors(KIR)gene polymorphism with bone marrow failure syndromes(BMFS). Methods SSP-PCR was used to examine the genotypic makeup of KIR in patients with aplastic anemia( AA), myelodysplatic syndrome (MDS) and healthy controls in our department. Results All the 16 KIR genes which had been prescribed were identified. The frequencies of KIR-2DS1, 2DS2, 2DS3, 2DS5 and 3DS1 genes were showed increased in patients with AA, MDS than in healthy controls. The patients with AA had lower frequency of KIR-2DS5 than the patients with MDS. Conclusion The increased frequencies of these activated KiRs in patients with MDS and AA suggest that the abnormal immunogenetic might be related to the pathogenesis of BMFS.

Objective To obtain and identify dendritic cell (DC) from mouse bone marrow in vitro. Methods Recombinant mouse granulocyte and macrophage colony stimulus factor (GM-CSF) induction was used to induce mouse bone marrow cells to form dendritic cells in vitro. The morphological changes were observed with light inverted microscope and scanning electron microscope (SEM). CD11c, MHCⅡ, and CD86 were identified with flow cytometry. The biological function was studied with antigen phagocytosis test and mixed lymphocyte reaction (MLR). Result The cultured DC from mouse bone marrow displayed the typical morphological characteristics of DC. Immature DC, which had high expression in CD11c and low expression in MHCⅡ and CD86, could phagocytize antigen. Mature DC, which had high expression in CD11c, MHCⅡ and CD86, could stimulate allogenic mixed lymphocyte proliferation. Conclusion DC can be generated from mouse bone marrow cells through cytokine induction in vitro and be used for further study associated with DC.

Objective To optimize the extracting conditions of polysaccharide from Imperata Cylindrica and establish a suitable method for its content determination. Methods Based on the qualitative yield of polysaccharide extraction, a orthogonal test was employed to evaluate the effect of four factors including extracting temperature, extracting time, the extraction times and the ratio of material to liquid. The phenol-dense sulphuric acid method was applied for the content determination of polysaccharide. Results The times of extraction and the extracting temperature influenced the content of polysaccharides mostly, while the duration of extraction and the amount of water showed little influence. Conclusions The optimum extraction condition was refluxing and extracting for 3 times at 85 ℃ for 3 hours, and with 15 folds of water for every time. The contents of polysaccharide in Imperata Cylindrica collected from two different places were above 0.15%.

Objective To investigate the effects of extractum trametes robiniphila murr combinated with hydroxyurea on proliferation and apoptosis of K562 cells and whether the 2 drugs have synergistic effects,and to detect the antiproliferation and apoptosis ratio of cells in diffe-rent groups were detected.Methods K562 cells were cultured in vitro,and logarithmic phase cells were used for study.MTT-inhibitory test and cell morphological analysis were employed to examine the effects of extractum trametes robiniphila murr alone or in combination with hydroxyurea on proliferation of K562 cells.The effects of extractum trametes robiniphila murr alone or in combinated with hydroxyurea on apoptosis of K562 cells were examined by flow cytometry.The expressions of bcr-abl,bax and bcl-xl mRNA levels were detected by way of reverse transcription polymerase chain reaction(RT-PCR).Results Extractum trametes robiniphila murr could inhibit proliferation and induce apoptosis of K562 cells,and extractum trametes robiniphila murr combinated with hydroxyurea had a synergistic effects on cell prolife-ration and apoptosis.The RT-PCR displayed that either extractum trametes robiniphila murr alone or in combinated with hydroxyurea could down-regulate the expression of bcl-xl mRNA and up-regulate the expression of bax mRNA and the effect of the 2 drugs had a synergistic effect on bcl-xl and bax mRNA expressions.Extractum trametes robiniphila murr alone could also down-regulate the expression of bcr-abl mRNA,but hydroxyurea alone did not show any effect on the expression of bcr-abl mRNA and the 2 drugs had no synergistic effect.Conclusions The effect of extractum trametes robiniphila murr combinated with hydroxyurea on the proliferation and apoptosis of K562 cells has a synergistic effect.The mechanism of antiproliferation and inducing apoptosis of extractum trametes robiniphila murr is probably related to down-regulating the expression of bcl-xl and bcr-abl mRNA and up-regulating the expression of bax mRNA.The synergistic effect of antiproliferation and inducing apoptosis of extractum trametes robiniphila murr combinated with hydroxyurea on K562 cells is probably related to down-regulating the expression of bcl-xl mRNA and up-regulating the expression of bax mRNA.

Objective To investigate the changes in trafficking of GluRl-containing AMPA (GluR1-AMPA) receptor and GluR2-AMPA receptor from cytoplasm to cell membrane in the spinal cord dorsal horn in a rat model of incisional pain.Methods Thirty-two adult male SD rats aged 6-8 weeks weighing 280-300 g were randomly divided into 2 groups:control group (group C,n =8) and incisional pain group (group Ⅰ,n =24).An 1 cm long incision was made in the plautar surface of right hindpaw according to Brennan et al.in group Ⅰ.Cumulative pain score (CPS) and paw-withdrawal threshold to yon Frey stimuli (PWT) were measured at 3 h and day 1 and 3 afar incision ( T1,2,3 ).The animals were sacrificed after pain behavior assessment.Their lumbar segments of the spinal cord (L3-6) were removed.The expression of GluR1 and GluR2 in cell membrane and cytoplasm in spinal cord dorsal horn was determined by Western blot analysis.The co-expression of Stargazing with GluR1 and GluR2 in the spinal cord dorsal horn was examined by co-immuno-precipitation.Results The CPS was increased and PWT decreased; the GluR1 expression in cytoplasm was decreased while the expression of GluR1 in cell membrane and the co-expression of Stargazing with GluR1 were up-regulated in group Ⅰ as compared with group C.There was no significant change in the expression of GluR2 in cytoplasm and cell membrane and the co-expression of Stargazing with GluR2 in group Ⅰ as compared with group C.Conclusion GluR1-AMPA receptor transfers from cytoplasm to cell membrane but GluR2-AMPA receptor does not in rats with incisional pain.

Abstract? AIM: To analysis the relation of severity of rhegmatogenous retinal detachment with the levels of amino acids and vascular endothelial growth factor ( VEGF) in the serum and in subretinal fluid.? METHODS: Forty -eight patients ( 52 eyes ) with rhegmatogenous retinal detachment treated in our hospital were selected.According to the degree of retinal detachment, patients were divided into <1/2 quadrant group, 1/2-3/4 quadrant group and>3/4 quadrant group. Fifty-five healthy objects for physical examination in our hospital were selected as the control group, to compare the differences of amino acids and VEGF levels in the serum.Correlation analysis on VEGF levels and amino acids in the serum and in subretinal fluid among patients with different grades of rhegmatogenous retinal detachment was conducted.? RESULTS: In patients with rhegmatogenous retinal detachment, the tryptophan in serum was 28.59±4.46mg/L, phenylalanine 8.95 ±2.55mg/L, methionine 8.15 ±2.17mg/L, valine 28.62 ±5.29mg/L, histidine 18.96 ±1.85mg/L and VEGF 589.92 ±185.34μg/L, which were higher than those in the control group, and the difference was statistically significant(P<0.05).The levels of phenylalanine was 9.85 ±1.21mg/L, histidine 20.63 ±2.07mg/L and VEGF 718.69 ± 283.34μg/L in the subretinal fluid of>3/4 quadrant group, which were significantly higher than those in the <1/2 quadrant group and 1/2-3/4 quadrant group ( P<0.05). VEGF in the subretinal fluid of VEGF in the rhegmatogenous retinal detachment group were positively correlated with phenylalanine (r=0.542, P<0.001), and histidine (r=0.782, P<0.001).?CONCLUSION: The levels of amino acids and VEGF in the subretinal fluid of patients with rhegmatogenous retinal detachment was higher than those in normals and increased with the severity of retinal detachment.