AIM: Respiratory tract cilia are difficult to harvest and observe due to weak transparence, which results from their location in thick mucosa, so people always try to obtain ciliated cells with many methods. This study was designed to apply manual microdissection to isolate ciliated cells from guinea pig's nasal epithelium and offer pure cells to further research. METHODS: The experiment was carried out at the Institute of Otolaryngology Head and Neck Surgery in Chinese PLA General Hospital from March of November in 2007. Twenty healthy white guinea pigs were recruited in this study. The ciliated cells layer was isolated from submucous layer by instruments for microsurgery under the light microscope. The isolated mucosa and untreated mucosa layer were stained with hematoxylin-eosin in order to verify the isolated layer is mucous ciliated cells layer. The protease-treated ciliated cells layer was viewed under low magnification microscope, to count number of single ciliated cells and impure cells, as well as size of ciliated cells group. The morphology of ciliated cells was observed under high magnification microscope. RESULTS: Result of hematoxylin-eosin stain confirmed that the cell layer by manual microdissection was ciliated cells and basal cell layer, which were thin without submucous layer. Under low magnification microscope after manual microdissection, the density of isolated ciliated cells was larger, number of single ciliated cells was more and size of ciliated cells group was smaller. There were about 7 ciliated cells in a cell group, and no other cells were observed. Under high magnification microscope, the isolated ciliated cells by manual microdissection were smooth, good in light refraction, and active for cilia beat. The survival time was prolonged obviously, still 50% cells were alive 24 hours later. Compared with conventional isolation technique, manual microdissection was significantly predominant in the purity and activity of cells (P

A real-time monitoring reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for the sensitive and specific detection of prototypic,prevalent North American porcine reproductive and respiratory syndrome virus (PRRSV)strains.As a higher sensitivity and specificity method than reverse transcription polymerase chain reaction (RT-PCR),the RT-LAMP method only used a turbidimeter,exhibited a detection limit corresponding to a 10-4 dilution of template RNA extracted from 250 μL of 105 of the 50% tissue culture infective dose (TCID50) of PRRSV containing cells,and no cross-reactivity was observed with other related viruses including porcine circovirus type 2,swine influenza virus,porcine rotavirus and classical swine fever virus.From forty-two field samples,33 samples in the RT-LAMP assay was detected positive,whereas three of which were not detected by RT-PCR.Furthermore,in 33 strains of PRRSV,an identical detection rate was observed with the RT-LAMP assay to what were isolated using porcine alveolar macrophages.These findings demonstrated that the RT-LAMP assay has potential clinical applications for the detection of highly pathogenic PRRSV isolates,especially in developing countries.

Objective To observe temporal expression of matrix metalloproteinases(MMP-13) and tissue inhibitor of metalloproteina-ses-1 (TIMP-1) in lung of newborn rats with hyperoxia induced chronic lung disease (CLD),and to explore the relationship of CLD with MMPs.Methods The neonatal rats within 24 hours after birth were randomly divided into hyperoxia-exposed group(n=40) and control group(n=40).On postnatal 1,3,7,14 and 21 days,lung tissue of rats in 2 groups were collected.Lung histological changes were evaluated by hematoxylin and eosin and Masson stain;Collagen Ⅰ was detected by enzyme linked immunoadsorbent assay;MMP-13 and TIMP-1 were identifide by immunohistochemistry.Results Exposured to hyperoxia enviroment for 21 days,the number of alveolar decreased,terminal air space enlarged,inter-alveolar septa thickened,and deposition of interstitial collagen fibers.On 14 and 21 days,collagen Ⅰ in the lung of hyperoxia-exposed group increased significantly compared with that of control group(P0.05),obviously decreased on 21 day(P

Asteroid Hyalosis (AH) is a common clinical disease,which has been considered a benign disorder as it rarely impairs visual acuity.It was often discovered when the patient was treated for other eye diseases.The mechanism was unclear.Its characteristic B-ultrasound property makes the B-ultrasound a very helpful diagnostic technique.In the case of the patients with other fundus diseases associated with AH,optical coherence tomography (OCT) and fluorescein angiography (FA) may be used to reduce the interference from asteroid bodies,therefore improve the fundus visibility.Recent studies have shown that AH can incorporate with many other eye diseases.For example,in patients with cataracts,asteroid hyalosis can cause surface calcification of silicone plate intraocular lenses,which in most cases may lead to the need for explantation of the calcified intraocular lenses.The efficacy of pars plana vitrectomy (PPV),the removal of some,or all,of the eye`s vitreous humor for AH remains controversial.In this paper,we provide a review of the recent literature on AH disease: the etiology,diagnosis and treatment.We hope to thus improve the awareness and outcomes of AH disease.

AIM: To investigate the expression and significance of hypoxia-inducible factor -1α (HIF-1α)and Cyclooxygenase-2(COX-2) in the retinal neovascularization.METHODS: Mouse model of hyperoxia-induced ischemic retinopathy were established. A novel flucorescein-dextran perfusion method has been developed to assess the vascular pattern. histological methods were used to count blood vessel profiles in the inner retina with HE staining.HIF-1α and COX-2 expression in the retina was determined by immunohistochemical method.RESULTS: Lots of neovascularization were seen in hyperoxia group. The number of nuclei of proliferative retinal vessels increased significantly as compared with normal control group (P＜0.001). The expression of HIF -1α was noted in the ganglion cells and retinal blood vessels. The expression of COX-2 was noted in the inner nuclear layer and ganglion cells and retinal blood vessels.CONCLUSION: There are the expression of HIF-1α and COX-2 in the retinal neovascularization and they have correlation.

BACKGROUND:Repeated hyperbaric oxygenation (HBO) preconditioning induces tolerance against ischemia in spinal cord. The confirmation of this phenomenon paves a new way in clinic to prevent and treat complications of spinal ischemia following thoracoepigastric aorta surgery. Probing into its mechanism of is chemic tolerance has provided basis for clinical application.OBJECTIVE: To investigate the changes of mitochondrial structure and ATPase activity during spinal cord ischemic tolerance induced by repeated HBO preconditioning in rabbits.DESIGN: Randomized and controlled trial.SETTING: Department of Anesthesiology, Xijing Hospital of Fourth Military Medical University of Chinese PLA.MATERIALS: The experiment was conducted in the hyperbaric oxygen chamber of the Department of Air Medicine; Experimental Animal Center of the Department of Anesthesiology, Xijing Hospital of Fourth Military Medical University of Chinese PLA, between March and June 2004. Totally 50 male New Zealand rabbits were selected.METHODS: Fifty male New Zealand rabbits were randomly divided into two groups: simple ischemia control group and HBO group with 25 rabbits in each group: 0.25 Mpa, 100% O2, one hour per day for 5 days. Spinal ischemia model was prepared 24 hours after the last preconditioning.Spinal cord ischemia (20 minutes)/reperfusion model was prepared by blocking the infrsrenal aorta. The motor function of hind-limbs was scored at 6 hours, 24 hours and 48 hours after reperfusion. The mitochondrial ATPase activity was measured and compared between two groups before ischemia, at 6 hours, 24 hours and 48 hours after reperfusion.MAIN OUTCOME MEASURES: ① The score of motor nerve function of the hind-limbs; ② The activity of mitochondrial ATPase; ③ Pathological evaluation.RESULTS: Totally 50 rabbits were involved for result analysis, and finally 25 rats in each group entered statistical analysis with no loss in the midway. ① The score of motor nerve function of the hind-limbs in HBO group at 6 hours, 24 hours and 48 hours after HBO preconditioning was significantly higher than that in control group (P ＜ 0.01). ② The activity of mitochondrial ATPase: the mitochondrial Na+, K+-ATPase activity and Ca2+, Mg2+-ATPase activity at each time point of the animals in the two groups after is chemia were significantly decreased; Na+, K+-ATPase activity in HBO preconditioning group at 6 hours, 24 hours and 48 hours of ischemia was significantly higher than that in control group (P ＜ 0.01); Ca2+, Mg2+-ATPase activity at 6 hours and 24 hours after ischemia was significantly higher than that in control group (P ＜ 0.01). ③ Pathological evaluation: The structure of mitochondria in control group was seriously damaged while it was found almost normal in spinal cord in HBO group animals at 48 hours after reperfusion.CONCLUSION: The mechanism of ischemic tolerance induced by hyperbaric oxygenation pretreatment may partly be related to the attenuation of mitochondria ATPase activity decrease after ischemia, thereby protecting the function of mitochondria.

Objective To measure the phase values of the deep gray nuclei by using susceptibility weighted imaging(SWI) in underage brains,and to investigate the correlation between the phase value and age.Methods A total of 105 healthy juveniles were examined in this study by using MRI conventional sequence and SWI.Their ages ranged from 0 to 18 years(0-＜ 1 year,60 cases; 1-＜3 years,10 cases; 3-7 years,10 cases; 7-12 years,10 cases; 12-18 years,15 cases).Phase values of deep gray nuclei were measured.The correlation between phase value and age was analyzed by Spearman correlation method.Results During 0-3 years,the phase value of caudate nucleus was the lowest within the same age group (0-＜ 1 year:left-0.0433 ± 0.0291,right-0.0369 ± 0.0215 ; 1-＜ 3 years:left-0.0369 ± 0.0215,right-0.0384 ± 0.0259),whereas the phase value of red nucleus was the highest (0-＜ 1 year:left 0.0286-0.0380,right 0.0254 ±0.0361 ; 1-＜3 years:left 0.0325 ±0.0237,right 0.0395 ±0.0270).After 3 years old,the phase value of globus pallidus was the lowest within the same age group (3-＜7 years:left-0.0967 ± 0.0656,right-0.0953 ± 0.0617; 7-＜ 12 years:left-0.1870 ± 0.0531,right-0.1724 ±0.0547; 12-＜ 18 years:left-0.2037 ±0.0492,right-0.1849-±0.0324),whereas the phase value of thalamus was the highest (3-＜ 7 years:left-0.0019 ± 0.0225,right -0.0007 ± 0.0167 ; 7-＜ 12 years:left-0.0067 ± 0.0104,right-0.0064 ± 0.0118 ; 12-＜ 18 years:left-0.0204 ± 0.0181,right-0.0172 ± 0.0133).During 0-18 years,a moderate negative correlation between phase values of bilateral caudate nucleus,globus pallidus,red nucleus,substantia nigra and age were observed (rs =-0.483,-0.497,-0.67,-0.621,-0.489,-0.43,-0.552,-0.58 respectively) A low negative correlation between phase values of bilateral putamen,thalamus and age were observed (rs =-0.272,-0.213,-0.382,-0.366 respectively).Conclusions There is a negative correlation between phase value and age in the deep gray nuclei of underage brains.And the phase value may be a useful index in the diagnosis of iron deposition related diseases.

The aim of this study is to establish a simple and stable model like poloxamer 407 (P-407)-induced dyslipidemia of golden hamster model, and investigate the mechanism of lipid metabolism disturbance in this model. PPARalpha agonist and HMG-CoA reductase inhibitor were administrated to validate the efficacy on regulating lipid metabolism in the dyslipidemia golden hamster model. Six weeks male golden hamsters were chosen to inject P-407 intraperitoneally at a bolus dose of 300 mg x kg(-1), an intermittent injection at a dose of 200 mg x kg(-1) every 72 hours after the bolus. The results showed that P-407-induced golden hamster model characterized as increased serum triglyceride (TG), total cholesterol (TC), free cholesterol (free-CHO), cholesteryl ester (CE), free fatty acids (FFA) and apoB levels, and the hyperlipidemia state maintained at a stable level persistently. Meanwhile, augmented malondialdehyde (MDA) and nitric oxide (NO) level was observed. LCAT and SR-B I mRNA levels in liver of model group were down-regulated (expression ratio is 0.426; 0.783), while HMG-CoA reductase mRNA level was up-regulated (expression ratio is 1.493) compared with those of the normal group. The serum cholesterol and triglyceride levels were significantly lower in P-407-induced dyslipidemia hamster model after treated with atorvastatin (Ato) at a dose of 50 mg x kg(1) or fenofibrate (Fen) at 100 mg x kg(-1) for two weeks. These findings suggest that serum lipid distribution in dyslipidemia golden hamster is similar to that of human, and which may be relevant to the disturbance of the enzymes expression involved in lipid metabolism in liver. Results obtained from this study support the concept that dyslipidemia golden hamster may be an adequate animal model to evaluate the efficacy of lipid-lowering agents.

Objective:To clarify the indications and efficacy after simultaneous surgical treatment in gastric cancer patients with synchronous hepatic metastases (GCHM).Methods:A total of 21 GCHM patients who underwent D2 gastrectomy with R0 hepatic resection (11 cases) or non-R0 hepatic resection (10 cases) from March 2004 to April 2016 were analyzed retrospectively.The clinicopathological characteristics and survival were compared between the two groups.Results:Cumulative survival rate was improved in R0 hepatic resection group compared with non-R0 hepatic resection group:1-year (54.5％ vs 48.0％),2-year(27.3％ vs 0％),and 5-year(27.3％ vs 0％,P=0.044).The median survival time in R0 hepatic resection group and non-R0 hepatic resection group were 16.2 and 5.9 months (P=0.008).Univariate analysis revealed that Bormann's classification (P=0.010) and state of regional lymph node (P=0.004) were significant predictive factors regarding cumulative survival rate.However,there was no significant prognostic factor (P=0.031) in multivariate analysis might partly owing to interaction among them and/or a small number of patients.Conclusion:Multidisciplinary treatments based on R0 hepatic resection combined with D2 gastrectomy could improve survival in selected GCHM patients.

Lidamycin (LDM) is a potent antitumor antibiotic. Previous studies have shown that LDM could inhibit proliferation and migration in endothelial cells. In the present report, the effect of LDM on angiogenesis of zebrafish embryo was studied. The results showed that treatment of zebrafish embryos with LDM resulted in significant inhibition of angiogenesis. Morphological observation, quantitative endogenous alkaline phosphatase (EAP) assay, alkaline phosphatase staining, and transgenic zebrafish assay were performed to evaluate vascular development defects in zebrafish. The results indicated that after the zebrafish embryos were exposed to LDM, angiogenesis defects of zebrafish embryos were observed, including pericardial edema, reduced numbers of circulating red blood cells, suppression of zebrafish vessel growth, and absences of SIV (subintestinal vein). The expression of VEGF was detected by RT-PCR assay, quantitative reverse transcriptase real-time PCR (qRT-PCR) assay and Western blotting analysis. The results revealed that LDM could inhibit the expression of VEGF protein, while the expression of mRNA was not significantly affected. The study suggests that LDM could inhibit the zebrafish embryo angiogenesis by down-regulation ofVEGF expression.