ObjectiveTo introduce a rapid and stable rat model of middle cerebral artery occlusion (MCAO). Methods32 Wistar male rats were used to induce focal cerebral ischemia according to Longa's method. The main difference from Longa's was to introduce a nylon suture into the cervical internal carotid artery without ligation the pterygopalatine artery (PPA). Regional cerebral blood flow (rCBF), neurological deficit, brain water content, red tetrzolium (TTC) stain and pathology were evaluated in all rats.ResultsThe operation proceeded about 15 min after anaesthesia. Rats after operation presented severe neurological deficits companied with rCBF decreases. There were significant increases of brain water contents in ischemic brain regions. TTC staining showed infarct areas and histological examinations revealed remarkable astrocyte swelling and neuronal necrosis.ConclusionInducing rat MCAO without ligation of PPA could produce reliable ischemic changes,with the advantage of shortening operation time.

Humans ; Organophosphorus Compounds ; analysis ; pharmacokinetics

Objective To investigate the protective effect of methylene blue (MB) on blood-brain barrier (BBB) injury after focal cere-bral ischemia-reperfusion in rats. Methods 18 male Sprague-Dawley rats were randomly divided into sham-operated group (n=6), model group (n=6) and MB treatment group (n=6). The left middle cerebral arteries were occluded for 1 hour and reperfused. MB was infused intra-venously immediately after reperfusion (3 mg/kg) and again 2 hours post-reperfusion (1.5 mg/kg), while normal saline was administered in the model group. The sham-operated group was treated as same as the model group without occlusion and infusion. HE staining was used to observe the histological injury in the cortex around the infarcted region 47 hours after reperfusion, while albumin immunohistochemistry was used to evaluate the permeability of the BBB, and immunohistochemistry and double immunofluorescence staining were used to exam-ine the expressions of glial fibrillary acidic protein (GFAP) and aquaporin-4 (AQP-4). Results HE staining showed that cells and blood ves-sels were not intact in the cortex around the infarcted region in the model group and they were better in the MB treatment group. The expres-sions of the albumin, GFAP and AQP-4 were higher in the model group than in the sham-operated group (P<0.01), and were lower in MB treatment group than in the model group (P<0.05). The double immunofluorescence staining showed the colocalization of GFAP and AQP-4 in the astrocytes. Conclusion MB may ameliorate the BBB disruption induced by focal cerebral ischemia-reperfusion through reducing glio-cyte proliferation and down-regulation of AQP-4 expression in rats.

Objective To analyze the relationship between age and the prognosis of patients with pancreatic neuroendocrine tumors (pNETs).Methods The clinical data of 102 patients with pNETs who were admitted to the Zhongshan Hospital of Fudan University from January 1999 to December 2010 were retrospectively analyzed.The properties of the tumors were determined by morphology and immunohistochemical staining of chromogranin A,synaptophysin and neuron-specific enolase.Preoperative grading of the tumors was done by mitotic count and Ki67 proliferation index,and the staging of the tumors was done by ENETS TNM system.All patients were divided into younger group (age≤60 years,77 patients) and older group (age ＞ 60 years,25 patients).The survival of the patients was estimated using the life table,and the survival curve was drawn by the Kaplan-Meier method.The survival of the 2 groups was compared using the Log-rank test.Multivariate analysis was performed with the COX proportional hazards model.Results The median survival time was 139.8 months.The overall 1-,2-,5-,and 10-year survival rates were 96％,93％,90％ and 80％,respectively.The postoperative survival time of patients in the younger group was significantly longer than that in the older group (x2 =4.717,P ＜ 0.05 ).The prognosis of patients with higher tumor grades ( G2,G3) and higher TNM stages ( Ⅲ,Ⅳ ) in the older group was significantly poorer than those in the younger group ( x2 =11.158,5.375,P ＜ 0.05 ).The results of multivariate analysis showed that age and major vascular invasion were the independent predictors for survival (RR =8.626,12.795,P ＜0.05 ).Conclusions Age above 60 years is an important independent factor influencing the prognosis of pNETs patients.Tumor grading and TNM staging are highly correlated with the prognosis of the pNETs patients.

Objective To investigate the clinicopathological features,treatment method and prognostic factors of the gastric neuroendocrine neoplasms (gNENs).Methods The clinical data of 80 patients with gNENs who were admitted to the Zhongshan Hospital of Fudan University from January 2002 to December 2011 were retrospectively analyzed.All the patients received gastroscopic examination.Patients with well-differentiated and diameter ≤ 2 cm gNENs received endoscopic mucosal resection or endoscopic submucosal dissection.Patients with poordifferentiated and diameter ＞ 2cm tumors received surgical resection of gNENs.Patients were followed up via phone call,mail or out-patient examination till October 31,2013.The Kaplan-Meier method was used for drawing the survival curve and calculating the survival rate.The univariate analysis was done by Log-rank test and the multivariate analysis was done by COX proportional hazards model.Results Fifty-eight patients had pain and discomfort in the epigastric region.Thirty-two gNENs were located at the cardia,40 at the body and 8 at the pylorus.Fortyfour gNENs were ulcerative type,25 were polypoid type,11 were protruded type.The mean diameter of the gNENs was 2.6 cm (range,0.4-7.5 cm).Twenty-seven gNENs were in grade 1,10 in grade 2 and 43 in grade 3.Forty-five gNENs were localized,34 gNENs had lymph node involvement (2 gNENs had distal metastasis),and 1 gNENs had distal metastasis.Thirty-one patients received endoscopic resection,and did not receive adjuvant therapy.Forty-nine patients (6 with gastric neuroendocrine tumor and 43 with gastric neuroendocrine carcinoma)received radical resection,including proximal subtotal gastrectomy in 16 patients,distal subtotal gastrectomy in 15 patients,total gastrectomy in 15 patients and distal subtotal gastrectomy + resection of the liver metastasis in 3 patients.Thirty-four patients received adjuvant chemotherapy and 15 did not receive adjuvant chemotherapy.Seventy-nine patients were followed up for a median time of 42.0 months (range,2.0-113.0 months).The mean time of survival was 75.6 months(range,2.8-100.8 months),and the 1-,3-,5-year overall survival rates were 91.3％,75.8％ and 66.5％,respectively.The 5-year survival rates of patients with gNENs in grade 1,grade 2 and grade 3 were 100.0％,100.0％ and 38.0％,respectively.The results of univariate analysis showed that the gender,treatment methods,adjuvant chemotherapy,types of tumor,tumor diameter,classification and staging of the tumor were correlated with the prognosis of the patients (x2=9.550,17.488,25.038,14.994,6.897,25.234,22.066,P ＜ 0.05).The results of multivariate analysis showed that the gender was the independent risk factor influencing the prognosis of the patients (RR =11.280,95％ confidence interval:5.353-19.121,P ＜ 0.05).Conclusions The clinical presentations of gNENs are often nonspecific.The main presentation of gNENs is pain in the epigastric region of the abdomen,and most of the gNENs are located at the cardia or body of the stomach.The staging and grading of the gNENs are varied,and the prognosis is related with the gender of the patients.Endoscopic or surigcal resection is the main treatment method for gNENs.Female patients have a better prognosis than male patients.

Objective To investigate the effects of interlukin-10 (IL-10) on lipopolysaccharide (LPS)-induced liver injury in rats.Methods One hundred male Wistar rats,aged 10-14 weeks,weighing 250-300 g,were randomly divided into 5 groups (n =20 each):control group (C group),LPS group,IL-10 group,HO-1 inducer cobalt protoporphyrin-Ⅸ group (Co group) and HO-1 inhibitor zinc protoporphyrin-Ⅸ group (Zn group).The animals in LPS,IL-10,Co and Zn groups received intraperitoneal LPS 20 mg/kg.IL-10,Co and Zn groups received recombinant human IL-10 1 μg at 3 h before LPS injection.Co and Zn groups received cobalt protoporphyrin-Ⅸ and zinc protoporphyrin-Ⅸ 25 mg/kg at 2 h before administration of recombinant human IL-10.Ten rats in each group were chosen at 24 h after LPS injection and blood samples were collected from the heart for determination of the levels of serum alanine aminotransferase (ALT),aspartate transaminase (AST),tumor necrosis factor-α (TNF-α) and interlukin-1β (IL-1 β).The animals were then sacrificed and lungs removed for determination of glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) activities,malonodialdehyde (MDA) content and expression of HO-1 mRNA in lung tissues.The left 10 rats in each group were chosen and the survival rates within 72 h after LPS administration were recorded.Results Compared with C group,the levels of serum ALT,AST,TNF-αand IL-1 β and MDA content in lung tissues were significantly increased,and GSH-Px and SOD activities in lung tissues and survival rates were decreased in LPS,IL-10,Co and Zn groups (P ＜ 0.05).Compared with LPS group,the levels of serum ALT,AST,TNF-α,IL-1β and MDA content in lung tissues were significantly decreased,GSH-Px and SOD activities in lung tissues and survival rates were increased,and the expression of HO-1 mRNA was up-regulated (P ＜ 0.05).Compared with IL-10 group,the levels of serum ALT,AST,TNF-α and IL-1β and MDA content in lung tissues were significantly increased,and GSH-Px and SOD activities in lung tissues and survival rates were decreased in group Zn (P ＜ 0.05),and no significant change in the parameters mentioned above was found in Co group (P ＞ 0.05).Conclusion IL-10 can attenuate LPS-induced liver injury in rats by inducing the expression of HO-1.

[Summary] Thyroid-associated ophthalmopathy ocular damage involved in lacrimal, tear film, cornea, and conjunctiva which can lead to ocular discomfort and decreased visual.The mechanism of thyroid-associated ophthalmopathy ocular surface damage and treatment is reviewed.

Objective To investigate the clinical features,recanalization after medical treatment and clinical outcome of cerebral artery dissection.Methods We reviewed the clinical records of ischemic stroke patients with cerebral artery dissection who visited the First Affiliated Hospital of Zhejiang University between October 2010 and December 2013.We recorded patients' general information,neurological deficit,imaging and clinical treatment.We followed up the patients and statistically analyzed demographic data,recanalization and clinical outcome.Results Among 28 cases of cerebral artery dissection,carotid dissection was the most common (n =19,67.9％),followed by vertebral artery dissection (n =7,25.0％),while the combination of carotid and vertebral artery dissection was rare (n =2,7.1％).In the imaging of cerebral artery dissection,wire beads and occlusion were the most common signs (n =15,53.6％),followed by rat tail sign (n =7,25.0％),aneurysmal dilatation (n =4,14.3％) and dualchamber levy (n =2,7.1％).There were 18 cases of acute cerebral infarction in the 28 studied cases,but there was no significant difference between the degree of stenosis and cerebral infarction caused by cerebral artery dissection.All patients received standard antithrombotic therapy.The difference of the recanalization rate between using anti-platelet aggregation and anticoagulant therapy was not statistically significant (5/6 vs 9/13,P =0.37).Conclusions Although cerebral artery dissection is relatively rare clinically,the typical clinical manifestations of the disease and the characteristic imaging are helpful for the diagnosis.Standardization of antithrombotic treatment is still the first-line treatment of cerebral artery dissection.

Objective To establish a stable and controllable model of acute regional cerebral ischemia in rats, and to evaluate it by CT perfusion imaging and histological study. Methods Twenty eight male Wistar rats were randomly divided into 4 groups, and there were 7 rats in each group. The sham operation rats were defined as the first group, rats suffered from cerebral ischemia for 15 minutes were classified as the second group, rats suffered from cerebral ischemia for 30 minutes and then reperfusion for 1 hour as the third group, and rats suffered from hypo perfusion for 6 hours as the fourth group. Cerebral ischemia or hypo perfusion were induced by inserting a nylon thread of different diameter into right middle cerebral artery (MCA) of rats under the monitoring of regional cerebral blood flow (rCBF) by the Laser Doppler Blood Perfusion Monitor (BPM). rCBF was also examined by dynamic CT perfusion imaging. At the end of the observation time, rats were decapitated, and three rats of each group were performed 2,3,5 triphenyltetrazolium chloride (TTC) staining and four rats were performed histological study. Results In the second group, rCBF was controlled within 5% to 22% under the monitoring by BMP and CT perfusion imaging showed the decreased rCBF in 7 rats, but TTC staining showed red appearance indicating no infarction focus formed. Electronic microscopic study revealed astrocytic swelling and a few of neuronal degeneration. In the third group, rCBF was controlled within 4% to 23% under the monitoring by BMP. There were more severe astrocytic swelling and a lot of neuronal degeneration. The abnormal areas in CT perfusion images were the same as TTC staining. In the fourth group, in accordance with less decrease ment of rCBF (from 38% to 55%) in 7 rats, there were obvious astrocytic swelling and subtle neuronal degeneration. TTC stain did not show ischemia area. All these abnormal changes were not observed in the sham operation rats. Conclusion The controllable acute regional cerebral ischemic model in rats is very stable and repeatable. It can be simulated into the ischemic state of different perfusion level. This model is suitable for the research of acute cerebral infarction and regional cerebral ischemia. The facts that parallel changes existed among BMP measurement, CT perfusion imaging, and brain histology indicated that CT perfusion imaging is accurate and sensitive in evaluating acute regional ischemia.

Objective To explore the effects of knocking down ULF gene on the apoptosis of non-small-cell carcinoma H1299 cell after treatment with etoposide.Methods Three ULF small interfering RNA(siRNA)sequences and one negative control siRNA sequence were designed and synthesized, and then individually transfected into H1299 cell via lentivirus.The interference efficiency of ULF-siRNA were screened by real-time PCR and Western blotting.Then the most target siRNA was used for apoptosis assay after treatment with etoposide,MTT assay for H1299 cell proliferation,flow cytometry for cell cycle distribution. Results The expression of ULF gene and its protein ULF were down-regulated in H1299 cell when transfected with ULF-siRNA,and ULF-siRNA-1 was the most effective one,which had the highest inhibition rate(80%)of ULF expression.Compared with negative control group,ULF-siRNA group showed an obvious apoptosis after treatment with etoposide,and the inhibition rate of was higher than control group,which was positively correlated with etoposide dose,the difference was statistically significant(P<0.05 ).Flow cytometry showed that compared with the control group,G0/G1 cell cycle in ULF-siRNA group was increased,and S phase cells was decreased,the differences were all statistically significant(P<0.05).Conclusion Down-regulation ULF protein expression through treatment with etoposide can induce apoptosis of non-small-cell carcinoma H1299 cells,and inhibit cell proliferation,which lead to cell cycle arrest.ULF gene may become the new target of gene therapy for cancer.