Objective To investigate the effect of grape seed proanthocyanidin extract (GSPE) on ultrastructure injury in hippocampous and cognition impairment in rat model of obstructive sleep apnea hypoxia. Methods 80 male Sprague-Dawley rats were randomly divided into control group, model group, high and low dose GSPE groups. The control group was exposed in air, while the model group was suffered from intermittent hypoxia conditions (50 ml/L, 8 h everyday, for 2 or 6 weeks), and the GSPE groups accepted GSPE 200 mg/kg or 100 mg/kg 2 weeks respectively before hypoxia. Pathology in hippocampal region was observed under electromicroscope. Malondialdehyde (MDA) contents and superoxide dismutase (SOD) activity were detected with colorimetry, and apoptotic cells were measured with TUNEL. The cognition function of rats was assessed with the Morris water maze (MWM). Results The ultrastructure in hippocampous was significantly injured,with the increase of MDA and decrease of SOD (P<0.001) in the model group. The apoptotic cells increased (P<0.001). The escaping latency prolonged (P<0.001) and the frequency of crossing the platform decreased (P<0.001) in MWM test in the model group. Compared with the model group, the GSPE groups decreased in MDA content, increased in SOD level, decreased in apoptotic cells and ultrastructure damages, shortened the escaping latency, and increased the frequency of crossing the platform (P<0.001), especially in the high dose group (P<0.05). Conclusion GSPE can relieve the damage of ultrastructure and improve cognition function after obstructive sleep apnea hypoxia in rats.

Objective To explore the mechanism of ischemic postconditioning in relieving cerebral ischemia reperfusion (IR) by regulating autophagy through P38MAPK pathway.Methods Cerebral ischemia reperfusion model was established by using modified Pulsinelli four-vessel occlusion (4-VO).Totally 128 male SD rats were divided into 4 groups randomly:control group (sham),cerebral ischemia reperfusion model group (CIR),cerebral ischemic postconditioning group (CIP),and cerebral ischemic postconditioning + P38MAPK inhibitor group (SB203580 group).Each group was subdivided into four time points:6 h,24 h,48 h,and 72 h.The morphological changes of the hippocampus CA1 area neurons at each time point and the number of surviving nerve cells were detected with HE staining.The expression of the hippocampus CA1 area phosphorylated P38MAPK and the autophagy-related genes of Beclin-1 and LC3-Ⅱ were detected with immunohistochemistry.The protein content of the hippocampus phosphorylated P38MAPK and autophagy-related genes of Beclin-1 and LC3-Ⅱ were detected with Western blotting.Results Compared with those in sham group,the damage of rats' hippocampal neuron structure and the survival rate of neurons at each time point decreased in CIR group,the expressions of p-P38MAPK,LC3-Ⅱ and Beclin-1 increased.Compared with those in CIR group,in CIP and SB203580 groups the structure of rats hippocampal neurons was improved,the survival rate of neurons increased,the expression of p-P38MAPK decreased and the expressions of LC3-Ⅱ and Beclin-1 increased at each time point.Compared with CIP group,SB203580 grouphad improved structure of rats' hippocampal neurons,increased survival rate of neurons,decreased expression of p-P38MAPK,and increased expressions of LC3-Ⅱ and Beclin-1 at each time point.Conclusion Cerebral ischemic postconditioning through inhibiting P38MAPK pathway can regulate autophagy and exert its nerve-protective effect.

Objective To study the effect of astragaloside on TGF-β1,SMAD2/3,and α-SMA expression in the kidney tissue of diabetic KKAy mice,and evaluate its potential role in renal interstitial fibrosis.Methods 20 type 2 diabetic KKAy mice were randomly divided into model group and astragaloside group,while 10 male C57BL/6J mice were selected as the control.Astragaloside at 40 mg · kg-1 · d-1 was given when the KKAy mice fed with high-fat diet to 14 weeks old.The mice in the control and model group received normal saline at 40 mg · kg-1 · d-1.Blood glucose meter was used to detect the blood glucose value of each mice at 16th,20th and 24th week.The mice were killed at 24 weeks old and the kidney tissue samples were collected.Pathology morphological changes were observed.Results (1) blood glucose value:cmpared with the control group,the blood glucose value of KKAy mice at 14 week increased significantly,and that of model group also increased significantly at 16th,20th and 24th week (P＜0.05);the blood glucose value of astragaloside group decreased compared with control group (P＜0.05).(2) Morphology of kidney:in the control group,the glomerular and tubular had clear structure,there was no renal interstitial fibrosis;in the model group,the renal glomerular mesangial matrix had broaden,mesangial cell had increased,renal tubular epithelial cell cytoplasm showed vacuole degeneration,renal interstitial inflammatory cell had increaised.In astragaloside group,there were few renal tubular epithelial cell cytoplasm,and there was no obvious fibrosis.(3)TGF-β1,SMAD2/3,and α-SMA expression levels of the kidney issuse:compared with control group,mice in model group up-regulated TGF-β1,SMAD2/3 and α-SMA expression (P＜ 0.05).TGF-β1,SMAD2/3,and α-SMA expression levels in astragaloside group were significantly lower than those in the model group (P＜0.05).There was few phosphorylated SMAD2/3 expression in renal tubular and glomerular nuclei,while that of model group increased (P＜0.01),and compared with model group,that of the astragaloside group decreased (P＜0.05).Conclusion Astragaloside can delay the renal fibrosis process in diabetic mice by influencing the TGF-β/SMADS signaling pathway and down-regulating TGF-β1 and α-SMA expression,thus to relieve renal fibrosis in diabetic mice.

Objective To establish a diagnosis method for fungal infection using two-round PCR,and to evaluate its sensitivity in the detection of clinical specimens suspected to be infected with fungi.Methods A total of 29 specimens of clinical sputum and alveolar wash solution were collected from patients with suspicious fungal infection.All specimens uaderwent direct microscopy with 10%KOH,fungal culture,one-round PCR and two-round PCR.The fungal universal primer targeting ITS regions of rDNA was used in PCR.The detection rate for fungi was compared between these methods.Results The detection rate for fungi was 20.69%by direct microscopy,37.9%by fungal culture,17.2%by one-round PCR,and 48.3%by two-round PCR.More than one species of fungus were detected in 6.9%(2/29),3.4%(1/29)and 24.1%(7/29)of these specimens by fungal culture.one-round PCR and two-round PCR, respectively.There was a significant difference in the detection rate between two-and one-round PCR(x2=6.34,P＜0.05).With regard to the detection rate for more than one species of fungus,two-round PCR was significantly higher than one-round PCR and fungal culture(x2=4.09,6.30.bom P＜0.05).Conclusion Two-round PCR may help to improve the sensitivity of molecular diagnosis of fungus-infected specimens.

Objective To investigate the application value of CT/MR image fusing in gross tumor volume (GTV) delineation of esophageal squamous cell carcinoma.Methods Twenty-nine patients with esophageal squamous cell carcinoma to be treated with radical surgery underwent routine CT scanning,MR T2-weighted imaging (T2WI) and diffusion weighted imaging (DWI) before surgery.Diffusion-sensitive gradient b values were taken at 400,600,and 800 s/mm2.GTVs were delineated on the CT image,CT/ MR T2WI,and CT/MR DWI respectively.The MR T2W1 image was used as the intermediary for the fusion of the CT image and MR DWI image.The length of GTVs measured under different imaging conditions were compared with the length of the resected specimen of esophagus.Results The GTV length was (44.94 ± 18.46) and (45.05 ±21.47) mm on the CT images and CT/MR T2WI images respectively.When the b values were 400,600,and 800 s/mm2,the esophageal carcinoma GTV length on CT/MR DWI images was (42.12 ± 17.79),(41.18 ± 17.17) and (39.77 ± 17.66) mm,respectively.The coefficient between the esophageal carcinoma GTV lengths on CT/MR DWI images and the pathological lesion lengths was 0.928,0.926 and 0.927 respectively.Conclusions CT/MR DWI images displays esophageal carcinoma GTV length more accurately,thus guiding the delineation of GTV effectively.

Objective To observe the effects of CO2 pneumoperitoneum on the enteric motor function and acetylcholine esterase(AchE) neuron and nitric oxide synthase(NOS) neuron in the enteric nervous system,and explore the neuromechanism of the CO2 pneumoperitoneum on renteric motor function.Methods Thirty-six Sprague Dawley rats were randomly divided into experiment group(n=24) and control group(n=12).The experiment group was divided into two subgroups namely pneumoperitoneum 30min group and pneumoperitoneum 60min group(12 each) based on the maintenance time of pneumoperitoneum.Rats in each group were gavaged with medicinal carbon powder,and then the transmission of carbon powder in small intestine was determined.The spreading specimens of intestinal myenteric plexus of small intestine were prepared and the stained AchE and NOS neurons were observed and compared.Results The propellant velocity of carbon powder was slower in pneumoperitoneum 60min group than that in pneumoperitoneum 30min group and control group(28.55%?3.45% vs 45.90%?6.30%,48.25%?5.28%,P0.05).The number of positive expression of AchE neurons in intestinal myenteric plexus decreased in pneumoperitoneum 60min group compared with that in pneumoperitoneum 30min group and control group(48.00?3.16 vs 58.82?4.62,61.83?4.17,P0.05).The number of positive expression of NOS neurons in intestinal myenteric plexus increased in pneumoperitoneum 60min group compared with that in pneumoperitoneum 30min group and control group(42.17?4.45 vs 32.50?4.34,30.83?3.6,P0.05).Conclusions Prolonged CO2 pneumoperitoneum can affect or damage cholinergic neurons and nitroxidergic neurons in the enteric nervous system to some extent,and it may be the underlying mechanism of the intestinal motor dysfunction after operation.

OBJECTIVE To investigate the epidemic situation of Acinetobacter in lower respiratory tract infection and their drug resistance,in order to provide evidence for clinical anti-infection therapy. METHODS The data of Acinetobacter from the sputum specimens of inpatients in our hospital with lower respiratory tract infection during 2006-2007 were collected and analyzed with the software the software WHONET5.4. RESULTS Among all pathogens in lower respiratory tract infection,Acinetobacter accounted for 9.2% in 2006 and 7.4% in 2007,the rate in deparment of neurosurgery,surgical ICU and respiratory ICU was higher. Acinetobacter had the highest susceptible rate to imipenem and were also susceptible to meropenem and cefoperazone/sulbactam. However,Acinetobacter had higher resistant rate to imipenem and meropenem while higher susceptible rate to cefoperazone/sulbactam in 2007 than in 2006. The susceptible rate of Acinetobacter to the third and forth generation cephalosporins,amikacin,levofloxacin and aztreonam was lower than 50%. CONCLUSIONS The drug resistance mechanism of Acinetobacter is so complicated that many kinds of drugs prove poorly effective. Carbopenems are recommended when single drug is utilized and drug combination based on the clinical and laboratory information can be tried.

Objective: To observe the difference in anti-inflammatory effect between living rhino horn and rhino horn by the method of comparative research to provide the experimental basis for the replacement of rhino horn by living rhino horn.Methods: The anti-inflammatory effects of living rhino horn and rhino horn were studied by the methods of paw edema in rats, cotton ball granuloma in mice, auricle swelling and peritoneal dye penetration.Results: Compared with that in the model control group, the foot metatarsus swelling degree at all time points in high (440 mg·kg-1) dose group and middle (220 mg·kg-1) dose group of living rhino horn and three doses groups of rhino horn showed statistical differences (P＜0.05 or P＜0.01).The high dose group (700 mg·kg-1) and middle dose group (350 mg·kg-1) of living rhino horn and rhino horn could significantly reduce the weight of cotton ball granuloma in mice (P＜0.05).Three doses groups (700, 350 and 175 mg·kg-1) of living rhino horn and rhino horn could significantly reduce auricle swelling in mice induced by xylene (P＜0.05 or P＜0.01).The absorbance of Evansan in the abdominal cavity in the middle dose group (350 mg·kg-1) of rhino horn and the high dose group (700 mg·kg-1) and middle (350 mg·kg-1) dose group of living rhino horn was significantly reduced (P＜0.05 or P＜0.01).There was no significant difference in the anti-inflammatory effect between living rhino horn and rhino horn at the same dose.Conclusion: Living rhino horn and rhino horn have a certain anti-inflammatory effect.The anti-inflammatory effect of living rhino horn is similar to those of the rhino horn, and living rhino horn can be used as a substitute of rhino horn.

Objective To explore the regulation of PI3K-mTOR signaling pathways on autophagy of hippocampus nerve cells after subarachnoid hemorrhage (SAH)in rats.Methods We randomly divided 72 male Sprague-Dawley rats into sham group,SAH model group and LY294002 group with 24 rats in each group.We established SAH model with the secondary injection of blood method while the sham group was not injected with blood.PI3K signaling pathways specific inhibitor LY294002 was injected with 500μmol per rat 30 minutes before modeling.After 6,24,72 and 144 h morphologic changes of hippocampus CA1 neural cells were observed by microscopy;the expression levels of PI3K,mTOR,Beclin-1 and LC3-Ⅱ were detected by immunohistochemical method.Results The density of survival neurons in the SAH group was significantly lower than that in the control group (P<0.05),PI3K-mTOR signaling pathways were activated obviously,and the expressions of Beclin 1 and LC3-Ⅱ were significantly higher than those in the control group (P<0.05 ).The number of survival neurons significantly decreased in the LY294002 group compared with the SAH group at each time point (P<0.05),PI3K-mTOR signaling pathways were suppressed.The expressions of Beclin-1 and LC3-Ⅱ were significantly lower than those in the SAH group (P<0.05).Conclusion PI3K-mTOR signaling pathways protect neurons by activating the autophagy of neurons after SAH.

Objective To investigate the changes in the expression of phosphatidylinositol 3?kinase(PI3?K),mammalian target of rapamycin (mTOR)and Beclin?1 in the hippocampus of normal rats and intermittent hypoxia rats with cerebral ischemia/reperfusion ,so as to explore the role of PI3K/mTOR/autophagy pathway in global cerebral ischemia/reperfusion injury aggravated by intermittent hypoxia. Methods A total of 80 healthy male Wistar rats were randomly divided into sham operation group(SO group,n=20),merely ischemia/reperfusion group(I/R group,n=20),intermittent hypoxia for 7?day ischemia/reperfusion group(IH7+I/R group,n=20),and intermittent hypoxia for 21?day ischemia/reperfusion group(IH21+I/R group,n=20). IH7+I/R group and IH21+I/R group were respectively given intermittent hypoxia for 7 days and 21 days before ischemia/reperfusion. The cerebral ischemia/reperfusion model was established by modified Pulsinelli four?vessel occlusion method. The morpholog?ical changes of nerve cells in hippocampal CA1 region were observed by HE staining and electron microscope. The protein expressions of PI3?K, mTOR and Beclin?1 of nerve cells in hippocampal CA1 region were detected by immunohistochemical staining and RT?PCR. The learning memory capacity of rats were assessed by the Morris water maze test. Results Compared with SO group,I/R group increased the never cells morphology damages,reduced the number of survival neurons,and declined the ability of learning and memory(P<0.05). Immunohistochemistry showed that the number of PI3?K immunoreactive cell,mTOR immunoreactive cell and Beclin?1 immunoreactive cell increased in I/R group compared with S0 group(P<0.05). RT?PCR showed that the expressions of PI3?K,mTOR and Beclin?1 increased in I/R group compared with S0 group(P<0.05). Compared with I/R group,intermittent hypoxia groups increased the never cells morphology damages,decreased the number of survival neu?rons,and declined the ability of learning and memory(P<0.05). Immunohistochemistry showed that the number of PI3?K immunoreactive cell, mTOR immunoreactive cell and Beclin?1 immunoreactive cell increased in IH7+I/R and IH21+I/R groups compared with I/R group(P<0.05). RT?PCR showed that the expressions of PI3?K,mTOR and Beclin?1 increased in IH7+I/R and IH21+I/R groups compared with I/R group(P<0.05),and the changes were more significant in IH21+I/R group(P<0.05). Conclusion Intermittent hypoxia can aggravate neurological injury after ischemia,which is related to PI3K/mTOR/autophagy pathway activation.