BACKGROUND:Periodontal tissue engineering technology provides new ideas and new ways for periodontitis-induced bone defect repair. OBJECTIVE:To develop a culture model for the periodontal ligament cells of miniature swine, which was constructed with hydroxyapatite, to investigate the biocompatibility with hydroxyapatite. METHODS:Periodontal ligament cells from miniature swine were harvested by using tissue explant method. Immunofluorescence was used to detect the expression of stromal cel antigen 1 in the periodontal ligament cells of miniature swine. The third passage cells were co-cultured with a three-dimension hydroxyapatite scaffold, and the biological characteristics of the cells were observed under a scanning electron microscope at days 1, 3, 7 of co-culture. RESULTS AND CONCLUSION:The pirmary miniature swine periodontal ligament cells grew wel , and they were positive for stromal cel antigen 1. Under the scanning electron microscope, the periodontal ligament cells of miniature swine grew wel on the hydroxyapatite scaffold at days 1, 3, 7 of co-culture. These prove that the miniature swine periodontal ligament cells, which can be separated using tissue explant method and cultured successful y in vitro, can grow wel on the hydroxyapatite scaffold.

Objective:To survey the effect of the Dureping Injection on the kinetic change of nitrous oxide(NO) in macrophage infected by the influenza virus FM1 strain.Methods:The murinal celiac macrophage(Ana-1) was infected by the virus,add the different concentration of the drug in the supernatant of the macrophage for 6 h,12 h,24 h and 36 h.The level of the NO in the supernatant was measured by the method of traditional Griess.Ana-1 cell line was infected by influenza virus FM1 strain,then treated with Dureping Injection 1 ?g/ml for 24 h.The cells were then collected,mRNA was extracted and the expression of NF-?B p65 was measured by RT-PCR;The nuclear protein was extracted and the expression of NF-?B p65 measured by Western blot.Results:60 ?g/ml,30 ?g/ml,10 ?g/ml and 1 ?g/ml group of Dureping Injection down-regulated amount of NO secreted by the Ana-1 cells infected by virus,and it was showed in dose relationship significantly;1 ?g/ml group of Dureping Injection down-regulated the expression of the NF-?B p65 mRNA and protein.Conclusion:Dureping Injection has an obvious effect against influenza virus FM1 strain by depressing the activity of NF-?B,which prevents the production of NO secreted by Ana-1 cells infected by the virus.

Objective To evaluate the changes in the expression of spinal aquaporin-4 (AQP4) during remifentanil-induced hyperalgesia in a mouse model of incisional pain.Methods Seventy-two pathogen-free healthy adult male CD1 mice,weighing 25-30 g,were divided into 4 groups (n=18 each) using a random number table:control group (group C),incisional pain (group I),remifentanil group (group R) and remifentanil plus incisional pain group (group R+I).Normal saline was infused subcutaneously in group C.An incision was made in the left hind paw in group I.Remifentanil 80 μg/kg was subcutaneously infused for 30 min at a rate of 0.8 ml/h in group R.Remifentanil was infused subcutaneously before establishment of the model in group R+I.The thermal paw withdrawal latency (TWL) and mechanical paw withdrawal threshold (MWT) were measured at 1 day before establishment of the model (T0) and 6 h and 1,2 and 7 days after establishment of the model (T1-4).After measurement of the pain threshold at T3,12 animals were sacrificed randomly,and the lumbar segment of the spinal cord was removed for determination of the distribution and expression of AQP4 by Western blot.Results Compared with group C,the TWL was significantly shortened at T1-3,and the MWT was decreased at T2-4 in R and R + I groups,and the expression of AQP4 was significantly up-regulated at T3 in I,R and R+I groups (P＜0.05).Compared with group I,the TWL was significantly shortened at T2,3,and the MWT was decreased at T2.4 in group R,and the TWL was significantly shortened at T1-3,the MWT was decreased at T2.4,and the expression of AQP4 was up-regulated at T3 in group R+I (P＜0.05).Conclusion The mechanism by which remifentanil induces hyperalgesia is related to up-regulation of AQP4 expression in the spinal cord in a mouse model of incisional pain.

MRDC is thefirst self-developing large integrated microbial resources database in the Chinese microbiological field. It includes Microbial Character Bank, Microbial Product Market Information Bank, Microbial Names Bank, Microbial Nomenclature Bank, RKC Code Bank for International exchange and one Microbiological Information Numerical Taxonomic System named MINTS.

Objective To evaluate the effects of remifentanil post-conditioning on aquaporin-1 (AQP-1) expression during myocardial ischemia-reperfusion (I/R) injury in rats.Methods Twenty-four male.SpragueDawley rats,weighing 250-300 g,were randomly divided into 3 groups (n =8 each) using a random number table:sham operation group (group S),group I/R,and remifentanil post-conditioning group (group RP).Myocardial I/R was induced by 45 min occlusion of left anterior descending branch of coronary artery followed by 24 h reperfusion.Remifentanil 10 μg· kg-1· min-1 was infused over 10 min starting from 10 min before reperfusion in group RP,while the equal volume of normal saline was given instead in S and I/R groups.At the end of reperfusion,all the rats were sacrificed and their myocardial specimens from left ventricles were obtained for microscopic examination of thepathological changes and for determination of AQP-1 mRNA (using real-time fluorescent quantitative PC R) and AQP-1 protein (by Western blot) expression in the ischemic area and myocardial water content.Results Compared with S group,myocardial water content was significantly increased in the other two groups,AQP-1 mRNA and protein expression was up-regulated in group I/R,and no significant change was found in AQP-1 mRNA and protein expression in RP group.Compared with I/R group,myocardial water content was significantly reduced,and AQP-1 mRNA and protein expression was down-regulated in RP group.Conclusion Remifentanil post-conditioning reduces myocardial I/R injury possibly through down-regulating AQP-1 expression in myocardial tissues of rats.

Objective To investigate the protective effects of insulin like growth factor 1(IGF-1) on cortical neurons under condition of hypoxia and the possible mechanism. Methods Cerebral cortical neurons from newborn rats were cultured under the condition of oxygen and glucose deprivation (OGD) . On day 7, neurons were treated with IGF-1 or IGF-1 plus LY294002 or PD98059 under condition of OGD or normal condition. MTT assay was used to analyze the viability of neurons in each group. The expression of total Akt and p-Akt were analyzed by Western blot. Results Compared with the control, the neuron viability was significantly higher in IGF-1 treated group under normal or OGD condition (P＜0.05). The protective effects of IGF-1 were attenuated in the presence of LY294002 but not PD98059. The result of Western blot showed IGF-1 upregulated the expression of p-Akt, which was inhibited by LY294002. Conclusion PI3K pathway may play an important role in neuroprotection afforded by IGF-1.

ObjectiveTo investigate the effects of sevoflurane anesthesia on aquaporin-9 (AQP-9) expression in brain tissue after focal cerebral ischemia-reperfusion (I/R) in rats.MethodsSeventy-five male SD rats weighing 230-270 g were randomly divided into 3 groups ( n =25 each):group sham operation (group S) ; group I/R and group sevoflurane anesthesia (group SE).All the animals were tracheally intubated under 2.0％ sevoflurane and mechanically ventilated.Anesthesia was maintained with fentanyl infusion at 25 μg· kg-1 · h-1 after a bolus of fentanyl 10 μg/kg and inhalation of 65％ N2O in O2 in groups S and I/R and with inhalation of 2％ sevoflurane in 35％ O2 in group SE.Focal cerebral ischemin was induced by occlusion of middle cerebral artery for 2 h using a nylon thread with rounded tip which was inserted into the right internal carotid artery and advanced cranially until resistance was met.The neurologic function was assessed and scored (0=no deficit,4 =unable to move,unconscious) and brain edema rate (volume of ischemic hemisphere-volume of contralateral hemisphere ÷volume of contralateral hemisphere × 100％ ) and expression of AQP-9 were determined at 6 h,1,2,3 and 5 d of reperfusion.ResultsFocal cerebral I/R significantly increased neurologic deficit scores,brain edema rate and AQP-9 expression in brain tissue in group I/R as compared with group S.Sevoflurane anesthesia significantly attenuated the I/R-induced increase in neurologic deficit scores and brain edema rate and further increased I/R-induced increase in AQP-9 expression in brain tissue.ConclusionSevoflurane anesthesia can reduce focal cerebral I/R injury by up-regulating the expression of AQP-9 in brain tissue.

Objeetlve To avoid inserting the tip of the central venous catheter into internal jugular vein (IJV) through subclavian vein under ultrasound guidance.Methods sixty breast cancel patients aged 28-63 yr weighing 41-70 kg who needed long-term intravenous infusion and chemotherapy through central venous catheter were randomly divided into 2 groups (n=30 each):control group (group C) and ultrasound group (group U).In group C the insertion of central venous catheter through subclavian vein was guided by pulsatile injection of ice-cold saline.In group U ultrasound detector (type HFL 38/13-6 MH,Sonosite Co,USA) was used to guide the insertion of tbe central vesons catheter.The position of the catheter tip was verified by X-ray radiography.The rate of successful placement at 1st attempt was calculated.Results The tip of the central venous catheter was correctly placed in the vena cava and right atrium in all patients in group U (success rate 100%),while in group C the tip was misplaced in IJV in 6 patients (success rate 80%) and had to be replaced.Conclusion Ultrasound guidance is effective for correct placement of the tip of central venous catheter in the vema cava and right atrium through subclavian vein.

Objective To observe the effect of ulinastatin on tumor necrosis factor(TNF-α)and interleukin-6(IL-6)in radiation-induced lung injury.Methods Severity-two female SD rats were randomly divided into 3 groups as control group,irradiation group and treatment group(administered with Ulinastatin).Rats in irradiation group and treatment group were irradiated with linear accelerator at a single dose of 25 Gy.After irradiation rats in treatment group were injected daily with ulinastatin at a dose of 100000 U-kg-1·d-1 for 7 days through caudal vein while rats in control group and irradiation group were injected with the same volume of saline.Rats were killed at 2 h,4,8 and 24 weeks.Samples of lung tissues were observed by using HE staining.Expression of TNF-α in lung was determined by Western blot and expression of IL-6 in serum was determined by ELISA.Data were analyzed by SPSS software.Results Expressions of TNF-α in lung and IL-6 in serum increased significantly after irradiated in irradiation group compared with control group,and it reached the peak at 4 weeks(q=5.63、6.21,P＜0.01).Though expressions of TNF-α and IL-6 in ffeatment group also increased compared with control group,the difference between irradiation group and treatment group was statistic significantly(q=4.97、7.42,P＜0.01).Conclusions TNF-α and IL-6 play an important role in radiation-induced lung injury.Ulinastatin could suppress the inflammatory response and radiation-induced lung injury effectively by decreasing the levels of TNF-α and IL-6.

BACKGROUND:Cerebral ischemia often occurs in underlying pathological conditions, such as hypertension,
hyperlipidemia and diabetes. Therefore, it is of great significance to construct a cerebral ischemia rat model with hyperlipidemia and to study the effect of basic pathological changes on the cerebral ischemia.
OBJECTIVE:To observe the brain tissue pathological changes of rat models with coexistence of hyperlipidemia and cerebral ischemia, and the effect of hyperlipidmia on cerebral ischemia.
METHODS:The rats were fed with high-fat diet to establish the hyperlipidmia models, and then focal cerebral
ischemia models were prepared with suture method. At 3 and 7 days after modeling, the 2,3,5-triphenyltetrazolium chloride staining was used to observe the volume of brain tissue ischemia, and hematoxylin-eosin staining was
performed to observe pathological change of the margin of the brain tissue ischemia zone.
RESULTS AND CONCLUSION:2,3,5-Triphenyltetrazolium chloride staining staining results showed that the volume of cerebral ischemia was significantly reduced in the hyperlipidemia+cerebral ischemia 7 day group. Hematoxylin-eosin staining results showed there was typical ischemic changes in al the cerebral ischemia models, and the number of microglial cel s after cerebral ischemia for 7 days was significantly smal er than that after cerebral ischemia for 3 days, and the changes were more obvious in the hyperlipidemia+7-day cerebral ischemia group when compared with the hyperlipidemia+3-day cerebral ischemia group. Ultrastructure showed there were neuronal and glial nuclear membrane shrinkage in al the cerebral ischemia models, mitochondria cristae was disappeared completely, endothelial cel mitochondria was decreased, most of the synaptic vesicles of nerve synapse were dissolved;the damages above were improved after ischemia for 7 days, especial y
hyperlipidemia+cerebral ischemia for 7 days, the neuronal degeneration and necrosis were reduced, the
mitochondrial damage was repaired, the number of mitochondrial cristae was increased significantly, and the synaptic vesicles of nerve synapse were recovered significantly. The results indicate that hyperlipidemia can promote the recovery of cerebral ischemic injury,
probably because the hyperlipidemia factors can activate the protection mechanism.