The changes of CaM content and distribution in acute cerebral ischemia of Mongolian gerbils were investigated with enzyme-linked immuno-sorbent assay (ELISA )and immunogold staining(IGS). The ELISA results showed that which ligation of both common carotid arteries for 10 rain the contents of CaM were significantly lower (P

The effects of bepridil, ni-modipine and nicardipine on the activity of CaM - dependent protein kinase Ⅱ from mouse cerebrum were studied in vitro. Tt was found that bepridil significantly inhibitied the activity of CaM - PK Ⅱ with an IC50 value of 0. 3 mmol ? L-1. but nimodipine and nicardipine did not basi-cally influence the activity of the enzyme. The experimental results showed that the inhibitory effect of bepridil on the activity of CaM - PK Ⅱ was an antagonistic action of bepridil to CaM.

Objective To study the molecular mechanism of glutamate receptor-6 (GluR6) in the pathogenesis of epilepsy. Methods Seizure model of SD rats was induced by intraperitoneal injection of kainate (KA). Immunoprecipitation and immunoblotting were performed to examine the interactions of GluR6 and MLK3 with PSD95 at various time points after KA injection. The effect of Tat-GluR6-9c on the MLK3 phospharylation induced by kainate was observed with immunoblotting and immunohistochemistry. Results The assembly of GluR6 and MLK3 with PSD95 was induced after KA hippocampal CA3 region, and bagan to decrease one day later. Pretreatment after KA injection in CA3 region (P<0.05). Conclusion KA induces the assembly of the GluR6-PSD95-MLK3 signaling module and subsequently activates MLK3, which ultimately results in brain injury.

AIM:To study the effect of human complement C 5b～9 complex on nitric oxide(NO) synthesis of glomerular mesangial cells (MC). METHODS: First, the human complement C 5b～9 complexes were isolated and glomerular MC of rats were cultured. Second, the MC were stimulated with C 5b～9 complex and changes of metabolism products of NO(NO 3 and NO 2) in MC culture supernatant at 6,24 and 48 hours after C 5b～9 stimulating were detected. Moreover, cGMP levels in cultured MC were also measured. RESULTS: NO 3/NO 2 contents from culture supernatant and cGMP levels in MC were increased parallelly after C 5b～9 complex stimulation. Further, NO synthesis was inhibited by L-NG-nitro-arginine-methylester(L-NAME). CONCLUSION: NO synthesis of rat glomerular MC was incerased by human complement C 5b～9 stimulation.

Objective To measure the organ weights , blood physiological and biochemical parameters of KK /Upj-Ay/J.Methods KK/Upj-Ay/J mice at five and ten weeks of age were selected , and the organ weights , blood physiological and biochemical parameters were observed .Results Parts of the organ weights , blood physiological and biochemical parameters of different ages and sexes were significant differences .The fasted blood glucose of KK/Upj-Ay/J mice reached 7.0mmol/L at 10 weeks of age .Conclusion The results show that the organ weights , blood physiological and biochemical parameters are affected by age and gender of KK /Upj-Ay/J mice.The fasted blood glucose reached the diabetes level at 10 weeks of age .

Objective:To compare the content of total saponins in Paridis Rhizome from Wudang mountain area to explore the cor-relation between the quality of medicinal materials and the production areas and species. Methods: The content of total saponins in Paridis Rhizome was determined by an ultraviolet-visible spectrophotometer at 406nm with perchloric acid as the chromogenic reagent. Results:The saponins content in Paridis Rhizome from Wudang mountain area had obvious differences:the minimum was 1. 29%, and the maximum was up to 10. 22%. The content of total saponins had no obvious correlation with species, production area and altitude. Conclusion:The quality of Paridis Rhizome is unstable in Wudang mountain area, and that will affect the effectiveness and safety of the clinical medication. Only by promoting the standardized planting of Chinese medicine materials, the stable quality of Paridis Rhizo-me can be ensured.

OBJECTIVETo introduce a safe and specific approach of (13)C magnetic resonance spectrum ((13)C MRS) spectroscopy and investigate the alterations in hepatic anabolism.

METHODSRelative anaplerotic, pyruvate recycling and gluconeogenic fluxes were measured by (13)C MRS isotopomer analysis of blood glucose from rats with 40% body surface area burn injury, and from rats exposed to sham injury. A short chain fatty acid, [U (13)C] propionate which was avidly extracted by the liver, was infused intravenously to deliver (13)C into the citric acid cycle. Proton-decoupled (13)C MRS of deproteinized plasma or extracts of the freeze-clamped liver were used to determine the distribution of (13)C in blood or hepatic glucose.

RESULTSThere was no difference in the multiplets detected in the glucose carbon-2 anomer from blood or liver after 45 or 60 minutes of the infusion of the propionate, indicating that steady-state isotopic conditions were achieved. Gluconeogenesis relative to citric acid cycle flux was not altered by burn injury; in both sham and burn groups the rate of glucose production was about equal to flux through citrate synthase. In the sham group of animals, the rate of entry of carbon skeletons into the citric acid cycle was about 4 times than that in the burn group. Similarly, flux through pyruvate kinase (again relative to citrate synthase) was significantly increased after the burn injury.

CONCLUSIONSSince results from analysis of the blood glucose are the same as that of the hepatic glucose, (13)C distribution in the glucose and hepatic metabolism can be assessed based on the (13)C MRS analysis of the blood glucose.

Animals ; Blood Glucose ; analysis ; Burns ; complications ; Carbon Isotopes ; Citric Acid Cycle ; physiology ; Disease Models, Animal ; Gluconeogenesis ; physiology ; Liver Diseases ; etiology ; pathology ; Liver Function Tests ; Magnetic Resonance Spectroscopy ; methods ; Male ; Probability ; Radiographic Image Enhancement ; Rats ; Rats, Sprague-Dawley ; Reference Values ; Sensitivity and Specificity

OBJECTIVETo observe the role of Kupffer cells in the postburn production of TNFalpha, IL-1beta and IL-6 in severely scalded rats.

METHODS(1) The production of TNFalpha, IL-1beta and IL-6 from rat Kupffer cells stimulated by burn serum was observed. (2) The postburn change in the expression of cytokine mRNA from rat Kupffer cells was monitored. (3) The change in the plasma cytokine contents in scalded rats was determined after the application of gadolinium chloride, a specific inhibitor of Kupffer cells.

RESULTSKupffer cells could be stimulated by burn serum to release cytokines TNFalpha, IL-1beta and IL-6. The mRNA expression of TNFalpha, IL-1beta and IL-6 from rat Kupffer cells increased significantly after injury. But the postburn plasma levels of TNFalpha, IL-1beta and IL-6 decreased obviously to 34.71%, 36.99% and 33.7% of those in scalding group, respectively, after the Kupffer cell activity was inhibited.

CONCLUSIONThe plasma cytokines, i.e. TNFalpha, IL-1beta and IL-6, were primarily produced from Kupffer cells after injury in scalded rats, initiated by TNFalpha, IL-1beta and IL-6 mRNA transcription.

Animals ; Burns ; immunology ; metabolism ; Gadolinium ; pharmacology ; Interleukin-1 ; biosynthesis ; genetics ; Interleukin-6 ; biosynthesis ; genetics ; Kupffer Cells ; physiology ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha ; biosynthesis ; genetics

OBJECTIVETo investigate the characteristics of the intra- and extra-hepatocyte sodium ions distribution in scalded rats during early postburn stage,with the aim of improving burn shock resuscitation regime and the resuscitation effects.

METHODSAdult Sprague-Dawley rats were randomly divided into sham scalding (C, n = 12) and scalding (S, n = 7) groups. The rats in S group were subjected to 40% TBSA III degree scalding on the back and were catheterized via jugular vein for fluid resuscitation. The rats in C group were catheterized via jugular vein without fluid infusion and were sham scalded by warm water in temperature of 37 degrees. The changes in the intra- and extra-hepatocyte sodium ion contents were determined in vivo by (23)Na-magnetic resonance spectrum technology, while the existing state of the intra- and extra-hepatocyte sodium ion was determined by detecting (23)Na-magnetic resonance horizontal delaying time (T(2)).

RESULTSThe extra-hepatocyte sodium content in S group at 24 postburn hours (PBHs) was 17% less than that in C group. In addition, the T(2f) (fast T(2)) in S group remained stable but maintained a higher ratio during the observation time. This suggested that the sodium binding sites in extra-hepatocyte matrix increased relatively and that intra-hepatocyte sodium content increased by 57%. But the T(2) and the fast and slow parts of the T(2) kept stable, which implied that intra-hepatocyte catabolizing products were increased. This led to an increase in the sodium ion binding sites within intra-hepatocyte matrix in proportion to the sodium ion content.

CONCLUSIONDuring early postburn stage, the extra-hepatocyte sodium in a remote organ such as the liver exhibited relative deficiency due to its ingress into hepatocyte cytoplasm and to the increase of sodium combining sites.

Animals ; Binding Sites ; Burns ; metabolism ; Hepatocytes ; metabolism ; Magnetic Resonance Spectroscopy ; Rats ; Rats, Sprague-Dawley ; Sodium ; metabolism

Objective:To observe the expression characteristics of eukaryotic translation initiation factor 2α(eIF2α), and analyze its proliferation regulation effect on fibroblasts of rheumatoid arthritis synovium.Methods:The synovial tissues were collected in patients with rheumatoid arthritis(RA)(40 cases) and osteoarthritis(OA)(40 cases). EIF2α and proliferating cell nuclear antigen(PCNA) were detected by immunohistochemistry method. Fibroblast cell line of rheumatoid arthritis synovium(MH7A) were cultured to establish si-eIF2α group(siRNA-eIF2α plasmid transfection), vector transfection group and blank control group in vitro. PCNA was detected by Western blot method, cell proliferation activity was detected by CCK-8 method. χ2 test was performed on count data, two-sample t-test was performed on quantitative data, one-way analysis of variance (ANOVA) was performed to compare the means of more than two groups, regression equation was calculated by correlation regression analysis. Results:The positive rate of eIF2α was significantly higher in RA synovial fibroblasts than that of OA [52.5%(21/4) vs 20.00%(8/20), χ2=9.14, P=0.003]. Positive correlation was found between eIF2α and PCNA in RA ( Y=0.366 X+2.220, P=0.001) . Compared with blank control group and vector transfection group, cell proliferation activity decreased significantly in si-eIF2α group of MH7A cell line at 72 h [(0.65±0.08) vs (0.96±0.12) vs (1.09±0.06), F=4.52, P=0.022] and 96 h [(1.13±0.14) vs (1.42±0.97) vs (1.56±0.12), F=9.87, P=0.001) , PCNA expression decreased significantly [(0.84±0.15) vs (1.32±0.18) vs (1.28±0.14), F=5.22, P=0.012) . Conclusion:High expression of eIF2α can promote the proliferation of fibroblasts of RA synovium.