OBJECTIVE:To develop a method for concentration determination of voriconazole in human plasma and apply it in the clinic. METHODS:UPLC-MS/MS method was adopted. Using ketoconazole as internal standard,the determination was per-formed on Shim-pack VP-ODS column with mobile phase consisted of water(containing 1‰ formic acid and 2 mmol/L ammonium acetate)-acetonitrile(gradient elution)at flow rate of 0.3 ml/min and column temperature of 40℃.The electrospray ion source,pos-itive ionizing pattern and multiple reaction monitoring were used;the mass transition ion-pairs of voriconazole and internal standard were m/z 351.2→282.2 and m/z 532.1→490.2. RESULTS:The linear range of voriconazole were 1-10 000 ng/ml (r=0.999 5,n=5),and the limit of quantitation was 1 ng/ml;RSDs of inter-day and intra-day were all lower than 10%;method recovery was higher than 90%(RSD<8%),and extraction recovery was higher than 70%(RSD<8%). The plasma concentrations of voriconazole in 10 patients with invasive fungal infection determined by this method were 507.33-7 011.24 ng/ml,and those of 3 patients were outside the recommended treatment concentration range. CONCLUSIONS:The established method is fast,accurate and sensitive,and can be applied for the therapeutic drug monitoring of voriconazole.

As a therapeutic strategy for ischemic stroke,neuroprotective agents are used to antagonize a series of harmful molecular biological events in cerebral ischemia.This article reviewes the current status of neuroprotective agents in the treatment of acute ischemic stroke,and the challenges from pre-clinical evidences translating into the clinical trials.The vascular endothelial cells,glial cells and neurons should be studied as a whole.

Iatrogenic gastrointestinal perforation is one of the severe adverse events of endoscopic therapeutic procedure. For acute iatrogenic perforation,management by endoscopic techniques is a simple and rapid modality to close the perforation with minimal invasiveness and avoiding the traditional surgical trauma. Endoclips,suture with special instruments,covered stents,degradable sheets combined with tissue adhesive,and combined endoscopic techniques such as snares combined with endoclips,are the major endoscopic therapeutic modalities for closure of iatrogenic gastrointestinal perforation. In this article,the current status and progress of endoscopic management for acute iatrogenic gastrointestinal perforation were reviewed.

Ear, Middle ; surgery ; Eustachian Tube ; Foreign Bodies ; surgery ; Humans ; Male ; Young Adult

Objective To examine outcomes of superficial vein stripping for vein varicosis caused by combination of superficial and deep venous reflux and the effect on deep vein reflux. Methods From January 2004 to December 2006, twenty patients (22 lower extremities) with combined superficial and deep venous reflux were enrolled for evaluation, patients with superficial venous reflux alone worked as controls.All patients underwent superficial vein stripping and followed up for 2 years. Venous clinic severity score (VCSS) and color duplex ultrasound test were used to evaluate the change of clinic symptoms and valve reflux of deep veins. Results Preoperative VCSS of superficial and deep vein reflux group and superficial vein reflux group was 7.1 ± 2. 9 and 6. 6 ± 2. 0 respectively (P = 0. 44). At two years after operation the VCSS was 2. 3 ± 1.2 and 1.8 ± 0. 8 respectively without significant statistic differences, whereas comparing postoperative VCSS with individual preoperative parameters there was a significant decrease (P <0. 01). In combined superficial and deep vein group, deep venous reflux reduced in 7 limbs, progressed in 4 limbs, no change in 12 limbs. While in superficial reflux alone group, deep venous reflux occurred in 4 limbs.Conclusions Superficial vein stripping effectively improves symptoms of great saphenous vein varicosis of lower extremity caused by combination of superficial and deep vein reflux without a significant effect on the stares of deep vein reflux.

Objective To investigate the expression of AIB1 protein in bladder cancer and its significance.Methods 30 cases of bladder cancer and 30 adjacent normal tissues were collected from 30 patients underwent radical or TURBt.The level of AIB1 protein expressed in these tissues were detected by immunohitochemical method.Results The positive rate of AIB1 protein in bladder carcinoma was higher than that in the normal tissues [43.3 ％ (13/30) vs 3.3 ％ (1/30),x2 =4.316,P ＜ 0.05].The positive rate of AIB1 in T3 stage was higher than that in T1 or T2 stage [62.5 ％ (5/8) vs 25.0 ％ (2/8),42.9 ％ (6/14),x2 =4.623,P ＜ 0.05].No correlation was found between the expression of AIB1 protein and the sex,age,multiplicity or pathological grade (P ＞ 0.05).Conclusions There is a close relationship between AIB1 and bladder cancer.AIB1 might have great value on the prognosis of patients with bladder cancer.

Objective To examine cerebrovascular reactivity to CO2 inhalation in mice. Methods In vivo Two-Pho?ton imaging technique was used to record the reaction of cerebral cortical vessels including penetrating artery, surface vein and capillary in 5 male C57 mice after CO2 inhalation under a thinned-skull cranial window. Nitric oxide syntheses inhibitor L-NAME and Prostaglandin syntheses inhibitor Indomethacin were used to block different vasodilator pathways, respectively. Results Different mouse cortical vessels displayed different degrees of dilation to 1-minute 5%CO2 inhala?tion. The penetrating artery exhibited the most obvious dilation (45.01%±4.45%). L-NAME intervention significantly di?minished cerebravascular CO2 reactivity(P<0.05). Indomethacin significantly attenuated the dilation of artery but not capillary comparing with L-NAME intervention(P<0.05). Conclusions Different vessels react differently to CO2 inhala?tion in which postaglandins and NO signal pathways are involved.

Objective To study Keshan disease and its developmental trend in Zhangjiakou City in 2010 and to provide a scientific basis for further prevention of the disease.Methods From the eight diseased counties of Zhangjiakou City,five diseased countries were selected; in each diseased county a diseased village was selected as the clinical investigating point.Physical examination and Electrocardiography (ECG) tracings were carried out to suspicious patients with Keshan disease.Suspected heart enlargement,and ECG abnormalities were taken anteroposterior chest X-ray to measure the cardiothoracic ratio.Results A total of 2519 people were examined,16 cases were diagnosed as chronic Keshan disease,79 as latent Keshan disease,and the detection rate was 0.64％ (16/2519) and 3.14％(79/2519),respectively.Total of 2519 ECG tracings were taken,and electrocardiogram abnormalities were 268 cases,with a abnormal rate 10.64％,and most of them were complete right bundle branch block[accounted for 17.16％ (46/268)].Totally 94 cases of anteroposterior chest X-ray films were taken and 42 cases were cardiac enlargement,with a detection rate 44.68％.The numbers of mild,moderate and severe cases were 20,12 and 10,respectively,and the constituent ratio was 47.62％ (20/42),28.57％ (12/42),23.81％ (10/42),respectively.Conclusions The incidence of Keshan disease in Zhangjiakou City is relatively low,and the condition is stable.But monitoring and comprehensive prevention and control measure still should be strengthened.

Objective To investigate the effects of insulin on vascular diameter of the peri -infarct region and infarct volume after cerebral infarction in mice. Methods Forty male C57/BL6j mice w ere randomly divided into a control group ( n = 5), a cerebral infarction group ( n = 15), a cerebral insulin resistance group (n = 5), and a cerebral insulin resistance infarction group ( n = 15). A model of cerebral infarction w as induced by the photochemical method. A model of cerebral insulin resistance w as induced by intracerebroventricular injection of streptozocin. Tw o -photon confocal microscope w as used to in vivo evaluate the changes of vascular diameter in the peri-infarct region at 20 min after insulin injection into the cerebelomedulary cistern. After modeling of cerebral infarction, artificial cerebrospinal fluid or insulin (10 ng/ml) w as immediately injected into the cerebelomedulary cistern, and the effect of insulin on cerebral infarct volume w as evaluated at 24 h after infarction. Results Insulin did not have significant effect on various types of cerebral vascular diameters in the normal control group, but it significantly contracted cerebral arteries ( -23.16% ±6.86% and -23.32% ±6.40%, respectively; al P <0.001) and penetrating arteries ( -15.20% ±5.51% and -16.40% ±4.27%, respectively; al P < 0.001) in the cerebral insulin resistance group and the cerebral insulin resistance infarction group, but it did not have any effect on the diameters of the cerebral veins. There w ere no significant differences in the vasoactive effects of insulin betw een the cerebral infarction group and the normal control group, as w el as betw een the cerebral insulin resistance group and the cerebral insulin resistance infarction group. Insulin significantly reduced the volume of cerebral infarction in the cerebral infarction group (9.0 ±1.0 mm3 vs.6.0 ±1.2 mm3; t = 4.294,P =0.002), and it did not have significant effect on the volume of cerebral infarction in the cerebral insulin resistance infarction group ( 12.6 ±2.3 mm3 vs.11.6 ±1.7 mm3; t = 0.782, P = 0.456). Conclusions Insulin can reduce ischemic brain injury in normal mice and can not affect the cerebrovascular diameter of the peri-infarct region. The neuroprotective effect of insulin is not significant in cerebral insulin resistance in mice, and it may be associated w ith the vasoconstrictor effects of insulin in the peri -infarct region.

BACKGROUND: Research has shown that the vascular endothelial growth factor (VEGF) of the ends of the fractured bone is heavily expressed 72 hours to 3 weeks after the fracture and it is supposed that it has a promoting effect on fracture healing. Inducing angiogenesis through VEGF gene transfection has gradually attracted the attention of the researches.OBJECTIVE: To find an efficient way of exogenous VEGF gene in vivo transfection through injecting the mixture of positive ion liposome transfection agent and plasmid and to study the promoting effect of extra VEGF gene expression on bone fracture healing.DESIGN: A randomly grouping, blank control trial.SETTING: Animal Laboratory of Huaxi Medical Center of Sichuan University MATERIALS: Totally 40 adult male SD rats, weighing 230 to 250 g,were involved. All the animals were randomly divided into the experiment group and the control group with 20 rats in each group.METHODS: The experiment was conducted at the Animal Laboratory of Huaxi Medical Center of Sichuan University from April to December 2003.Altogether 40 rats were involved to establish fractured models of right shaft of femur. Cut the bone in the middle of bone stem, retroplanted a Kirsh' nail with 1 mm diameter through intercondylar part and the fractured bone was fixed. In the experimental group, a mixture of 100 μL of liposome transfection agent and 100 μg of pBLAST49-mVEGF plasmid was injected in multiple points into the subperiosteum of the both sides of the ends of the fractured bone. The same volume of normal saline was injected into the rats in the control group. Then, 2 rats in each group were put to death 3,7,14,28,42,56,70 days after the operation and femoral bone specimen was collected.MAIN OUTCOME MEASURES: Observation of right femoral fractured staining results of VEGF, with the apperance of brown granules as positive.RESULTS: Two rats were selected at 7 time points separately, and altogether 28 rats entered the stage of result analysis. The other 12 rats were fracture at different time points: For the experimental group, 28 days after the operation, cartilage callus appeared and replaced fibrocallus gradually,and the fracture line disappeared. Fifty-six days after the operation, the bone healed completely. For the control group, 28 days after the operation , fibrocallus was observed, and the fracture line was still clear. 56days after the operation, much callus appeared, and the fracture line beof fractured bone was stained with hemotoxylin eosin (HE). In the experiment group, 56 days after the operation, the bone healed completely and trabecular like bones were rebuilt. The bone marrow cavity of the fractured region was open again. In the control group, Fifty-six days after the operation, no mature bone was formed, and the bone marrow cavity was not different time points: The expression in the two group reached to the peak on day 14 and began to decrease on day 28. The expression of VEGF in the experimental group was obviously higher than that in the control group.CONCLUSION: Injection of the mixture of positive ion liposome transfection agent into the subperiosteum of rats is an effective approach for in vivo transfection and pBLAST49-mVEGF gene transfection can effectively facilitate the bone fracture healing of rats.