Objective To evaluate the therapeutic effect and mechanism of specific inducible nitric oxide synthase(iNOS)aminoguanidine(AG)in rats with obstructive jaundice.Methods Forty male Wistar rats were divided randomly into four groups: normal control group, sham operation group, obstructive jaundice group and aminoguanidine therapeutic group.Each group had 10 rats.We assayed levels of liver function,hemobilirubin, endotoxin,lactic acid and malondialdehyde before and after therapy, and we also analyzed pathology of the liver and small intestine.Then we could explore the therapeutic effect of AG in rats with obstructive jaundice.Results The levels of endotoxin,lactic acid and malondialdehyde in blood increased progressively along with the pathological changes of the liver and small intestine.Each of the AG group parameters was significantly lower, and the pathological changes of liver and small intestine were improved.Conclusion AG could protect liver and small intestine by attenuating lipid peroxidative and endotoxemia,and provide a new way to cure obstructive jaundice.

OBJECTIVE:To investigate the first-pass effect and mechanism of vitexin-4′-O-glucoside(VG)in rats so as to pro-vide a basis for new drug development. METHODS:10 SD rats were divided into a group of hepatic portal venous administration and a group of femoral venous administration,which respectively received VG iv at superior mesenteric vein and femoral vein,and then metabolic rate was calculated by finding out the AUC of VG in the rats’livers. 15 SD rats were divided into a group of gastric infusion,a group of intestinal infusion and a group of hepatic portal venous infusion,which respectively received VG by infusion at gastric fundus and duodenum and iv at superior mesenteric vein,and then metabolic rate was calculated by finding out the AUC of VG in the rats’stomachs and intestines. 15 SD rats were divided into a group of intestinal infusion,a group of femoral venous administration and a group of normal saline. At 10 min before administration,the former two groups were given by infusion vera-pamil injection(60 ml/kg),the substrate of CYP3A and P-glycoprotein(P-gp);and the group of normal saline were given by infu-sion of isometric normal saline,and then the rats were given VG as above to observe the effect of verapamil on intestinal absorp-tion of VG. RESULTS:The metabolic rates of VG in the liver,stomach and intestine were 54.9%,1.7% and 91.9% respectively. After infusion of verapamil,slight increase in AUC of VG was found in the rats in the group of intestinal infusion. CONCLU-SIONS:The first-pass effects in the liver and intestine are the main factors related to the low bioavailability of VG. Based on pre-liminary judgment,VG is the substrate of intestinal CYP3A and/or P-gp.

Objective To study the effect of Src tyrosine kinase inhibition on subcutaneously transplanted tumor of human lung adenocarcinoma in mice and its mechanism. Methods For the subcutaneously transplanted tumor model, A549 cells or PC-9 cells were inoculated into SCID mice by subcutaneous injection. Immunohistochemistry was used to show the effect of Src tyrosine kinase inhibition on proliferation index (Ki-67 staining) and microvessel density (CD31 staining) of subcutaneously transplanted tumor of human lung adenocarcinoma in mice. Results Subcutaneously transplanted tumor of PC-9 cells was sensitive to src tyrosine kinase inhibitor. There was significant difference between treatment group and control group (P <0.01). There was significant difference between the two treatment group too (P <0.01). Stopping treatment for 1 week, the inhibition rate of tumor growth were 33.19 % and 84.79 % in 10 mg·kg-1·d-1 and 50 mg·kg-1·d-1 treatment group, respectively. The same treatment was less effective to subcutaneous tumors produced by A549 cells. Treatment with 50 mg·kg-1·d-1 Src tyrosine kinase inhibitor significantly reduced the proliferation index of subcutaneously transplanted tumor produced by PC-9 cells (P<0.01) and tended to reduce the proliferation index of subcutaneously transplanted tumor produced by A549 cells (P >0.05). Treatment with 50 mg·kg-1·d-1 Src tyrosine kinase inhibitor significantly reduced micro vascular density in both PC-9 and A549 induced subcutaneous tumors (P <0.05). Conclusion Inhibition of Src tyrosine kinase could suppress the progression of subcutaneously transplanted tumor, not only by the inhibition of cell proliferation of lung adenocarcinoma cells directly, but also by the inhibition of angiogenesis indirectly.

AIM:To investigate the effect of NF-κB binding element deletion on transcriptional regulation of (NOX1).METHODS:pGL3-Basic vector inserted with the NOX1 proximal promoter, and the same vector inserted with the NOX1 proximal promoter in the absence of the positive NF-κB binding element, were constructed.After cloning, diges-tion and purification, NOX1 proximal promoter (≈1 415 bp) was inserted into the multicloning site of the pGL3-Basic vec-tor and then sequenced ( pGL3-NOX1-1415) .NF-κB binding elements in the NOX1 promoters were predicted by Alibaba 2.1 software.The positive element was deleted by overlapping PCR.The deletion mutant was inserted into the pGL3 vector in the same way (pGL3-NOX1-1327).The plasmids were transfected into A549 cells, and then the cells were stimulated with TNF-α.The luciferase activity was monitored on MD SpectraMax M5 enzyme-labeled instrument.RESULTS:The se-quences of pGL3-NOX1-1415 and pGL3-NOX1-1327 were identified to be correct.Compared with control group, the lucif-erase activity was significantly higher in the cells transfected with pGL3-NOX1-1327 (P<0.05), but it was significantly lower than that in the cells transfected with pGL3-NOX1-1415 (P<0.05).CONCLUSION: NF-κB plays an essential role in the transcriptional regulation of NOX1 in TNF-α-induced A549 cells.Activated NF-κB binds to specific elements in NOX1 promoter regions to control the transcription.

Objective To investigate the therapeutic effect of frozen-thawed murine islets which were transplanted into diabetic rats after cultured with hyperbaric oxygenated rotary cell culture system (HORCCS). Methods The purified rat islets were divided into two groups: A. In vitro experiment groups (IvEG) : The rat islets in each subgroup were cultured in HORCCS or common medium for 30 days, then evaluated for the intracellular DNA and insulin contents of islets, and the viability and insulin secreting level of islets. B. Islet transplantation experimental groups (TxEG) : The frozen-thawed islets were cultured in HORCCS or common medium for 7 days, and then transplanted into the recipients. We observed the blood glucose level (BGL) and insulin secreting level in the recipients as well as the uhrastructure change of islets in TxEG. Results The viability and insulin secreting level of islets cultured with HORCCS at 14th day were much higher than those cultured with common medium (P <0.05). The blood glucose level in recipients transplanted with islets cultured with HORCCS recovered to normal value at the 2nd week and lasted for 8 weeks. All these recipients maintained the normal glucose tolerance curve. Electronic microscopy found microchannel outlets on the surface of the frozen-thawed islets cultured with HORCCS. Conclusions Frozen-thawed islets cultured with HORCCS could establish nutrient transmission microchannels, which were not only capable of oxygen and nutrients transmission, but also improving cryopreservation solution to diffuse inside the islet cells evenly and uniformly. So this method not only lessens islet damage from cryopreservation, but also improves the effect of transplantation.

Objective To research the value of bed fiber bronchial microscope technology ha neurosurgical ICU. Methods 28 cases in neurosuxgical ICU who had received bed fiber bronchial microscope operations,including blood and vomit aspiration, bronchoalveolar lavage, sputum bolt clearance were retrospectively analyzed. Results 4 cases of blood and vomit aspiration was cleaned ha time,improvement of respiratory function,31 cases was sputum as- piration by bed fiber bronchial microscope and bronchoalveolax lavage, 31 cases was sputum collected by fiber bronchial microscope and positive rate is 100% ,4 cases of atelectasis was lung recruitment by fiber bronchial micro- scope sputum aspiration. Conclusion Bed fiber bronchial microscope technology can eliminate waste effectivdy, clean dust,there axe great value in treatment of neurosurgical ICU patients with respiratory diseases.

BACKGROUND:Endogenous hydrogen sulfide can be used as a new gaseous signaling molecule, and has important signal transfer function and biological regulation effects. OBJECTIVE:To study the neuroprotective effects of hydrogen sulfide in rats with acute cauda equina syndrome. METHODS: The 72 Sprague-Dawley rats were randomly assigned to three groups. Experimental group, model group: laminectomy was performed at the lumbar 4 (L4) level of the vertebra, and a piece of silicone (10 mm long, 1 mm thick, and 1 mm wide) was placed under the laminae of the L5-6 vertebra to produce the animal model of cauda equina syndrome. Sham surgery group: a simple laminectomy was performed in L4, but silicone was not implanted. In the experimental group, 20 μmol/kg NaHS was injected intraperitonealy at 1 hour before model establishment. Model and sham surgery groups: an equal volume of saline was injected intraperitonealy. At 12, 24, 48 and 72 hours after model establishment, malonaldehyde and glutathione levels in cauda equina nerve tissue were detected. Simultaneously, hematoxylin-eosin staining and TUNEL staining were performed at 48 hours. RESULTS AND CONCLUSION:Hematoxylin-eosin staining demonstrated that cauda equina nerve tissue was dense and regular, with complete myelin sheath, no axon sweling in the sham surgery group. Cauda equina nerve tissue was sparse, with the presence of demyelination, and partial axons and myelin sheath sweling in the model group. Cauda equina nerve tissue was tight, with axonal sweling and demyelination in the experimental group. TUNEL staining demonstrated that the number of positive cels was less in the spinal cord and dorsal root ganglia in the sham surgery group. Abundant positive cels were detected in the spinal cord and dorsal root ganglia in the model group. The number of positive cels was significantly lower in the experimental group than that in the model group. Malonaldehyde levels were lower in the sham surgery and experimental groups than in the model group (P < 0.05, P < 0.01), but glutathione levels were higher than model group (P < 0.05,P < 0.01). These results indicated that hydrogen sulfide could decrease oxidative stress and protect cauda equina nerve in rats with acute cauda equina syndrome.

Objective To investigate the correlation between cytomegalovirus infection and heart rate variability (HRV) in elderly patients with atherosclerosis.Methods 160 patients with coronary heart disease who met World Health Organization diagnostic criteria for coronary heart disease in 1979 were collected.According to the IMTHCMV PP65 antigen test results,patients were divided into positive group (observation group,n=103) and negative group (control group,n=57).We detected the levels of of HRV,metalloprotease-9 (MMP 9) and tumor necrosis factor α (TNF-α) in the two groups in order to access the plaque stability.Results The all sinus standard deviation of RR interval (SDNN),standard deviation of the average NN interval (SDANNI),mean value of sinus standard deviation of RR interval (SDNNI) were lower in observation group than in control group (P ＜0.05).There were no significant differences in square root of the mean squared differences of successive NN intervals (RMSSD) level and percentage of differences exceeding 50ms between adjacent normal number of intervals (PNN50) between the two groups (P＞0.05).The levels of total cholesterol and low density lipoprotein were higher and the levels of MMP 9 AND TNF α were lower in observation group than in the control group (all P＜0.05).Compared with control group,the plaque stability was decreased in observation group [20.4％ (21/103) vs.61.4％ (35/57),x2=4.273,P=0.015].Conclusions Patients with atherosclerosis combined with cytomegalovirus infection have a greater heart rate variability and poorer plaque stability.

BACKGROUND:Under hypoxic environment, hypoxia-inducible factor 1α plays a dualregulatory role in cel apoptosis. Severity of hypoxia is the key to determine whether cels appear to have apoptosis or adapt to survive. When the cels are exposed to chronic or extreme hypoxia, a lack of protection mechanisms from hypoxia-inducible factor-1α can induce cel apoptosis. OBJECTIVE: To research the expression of hypoxia-inducible factor 1α in human lumbar nucleus pulposus of different herniated types and its relationships with cel apoptosis. METHODS: The nucleus pulposus was harvested from 60 cases of herniation of lumbar intervertebral discs, L4-5 in 41 cases and L5-S1 in 19 cases. The nucleus pulposus tissues were equaly divided into protruded and sequestered groups. Meanwhile, the nucleus pulposus tissues from another 10 cases of lumbar spine fracture were taken as control group. Expression of hypoxia-inducible factor 1α and apoptosis of lumbar nucleus pulposus cels were observed and detected with immunohistochemical technique and TUNEL method. Correlation of hypoxia-inducible factor 1α and apoptosis in human lumbar nucleus pulposus of different herniated types was analyzed. RESULTS AND CONCLUSION: The expression of hypoxia-inducible factor 1α was visualized in each case, but it was significantly higher in the sequestered group than in the protruded group and control group (P < 0.01). Apoptosis of nucleus pulposus cels were found in al the three groups, but the apoptotic rate was also higher in the sequestered group than in the protruded group and control group (P < 0.01). Expression of hypoxia-inducible factor 1α was positively correlated with cel apoptosis in human lumbar nucleus pulposus (P < 0.01). Overal, the expression of hypoxia-inducible factor1α in degenerative human lumbar nucleus pulposus is associated with herniated types, which is the highest in the sequestered type. The relationship between hypoxia-inducible factor 1α and apoptosis is positive.

BACKGROUND:Under hypoxic environment, hypoxia inducible factor-1 plays an important role in regulation of hypoxia-induced gene expression in the intervertebral disc. Hypoxia-inducible factor-1 consists of α and βsubunits, and which hypoxia inducible factor-1α determines the stability and activity of hypoxia-inducible factor-1. OBJECTIVE: To observe the expression of hypoxia inducible factor-1α in the human lumbar nucleus pulposus of different herniated types and to judge their relationships. METHODS:A total of 60 nucleus pulposus samples were harvested from the lumbar vertebra, including 41 from L4-5 and 19 from L5-S1, and then divided into protruded group and sequestered group, with 30 cases in each group. Meanwhile, another 10 samples of lumbar nucleus pulposus served as controls. Hematoxylin-eosin staining and streptavidin-biotin peroxidase complex immunohistochemical technique were used to observe the expression of hypoxia inducible factor-1α in the human lumbar nucleus pulposus in different groups. RESULTS AND CONCLUSION:The expression level of hypoxia inducible factor-1α was (58.2±7.5)% in the sequestered group, (27.3±2.3)% in the protruded group, and (10.5±4.7)% in the control group, which was significantly higher in the sequestered group than the other two groups (P < 0.01). These findings indicate that the expression of hypoxia inducible factor-1α in the lumbarnucleus pulposus is associated with the herniated types, which is the highest in the prolapse sequestered type.