AIM: To study the significance of expressio n of fms-like tyrosine kinase 4 (Flt-4) in different histopathological grades of a strocytoma. METHODS: The surgical specimens from 50 brain astrocytoma patien ts were stained immunohistochemically for examining Flt-4 and vascular endotheli al growth factor (VEGF) expression. Intratumoral microvessel density (IMVD) was calculated by labeling the endothelial cells of the blood vessels within the tum or. RESULTS: Flt-4, VEGF expression were closely correlated with his topathological grades of astrocytoma. Flt-4 and VEGF expression were found in 52 % (26/50), 60% (30/50) of tumors. A significant correlation was found between Fl t-4 and VEGF expression (P

?AIM: To discuss the application of anterior segment optic coherence tomography ( AS-OCT ) in the diagnosis of corneal ulcer.?METHODS: The cross linear scan was used in 88 patients ( 88 eyes ) with corneal lesion by AS-OCT to gather the image data, observe the pathological changed tissue by measuring all layers for patients with initial inspection, providing important visual images and data for treatments. All the patients were followed up for 2mo.?RESULTS:Clear images with structure of all layers were obtained. It can provide the intuitive image data and scan the same position and show the changes during the treatment.?CONCLUSION: AS-OCT can discover the important condition immediately. It also can monitor course of disease dynamically, provide the intuitive image data for clinical treatment.

Air ; analysis ; Air Pollutants, Occupational ; analysis ; Humans ; Occupational Exposure ; Phthalic Acids ; analysis ; Spectrophotometry ; methods ; Workplace

OBJECTIVETo present an improved method to obtain pure, viable, freshly isolated hepatic stellate cells.

METHODSAdult male SD rats were used. All procedures were performed with the animals under sodium pentobarbital anesthesia. Three days after the single intravenous administration of 1 ml liposome-encapsulated CL2MDP, which has selective cytotoxicity of Kupffer cells, livers were perfused with D-Hank's solution containing 100 U/ml heparin for 10 to 15 minutes, and then with 0.05% collagenase dissolved in D-Hank's solution for 25 to 30 minutes. The liver was then gently homogenized and further incubated in 0.025% collagenase, and 0.005% DNAase I for 30 minutes at 37 degrees C under constant stirring. This suspension was filtered through stainless steel gauze and centrifuged for 2 minutes at 50 x g to remove parenchymal cells. Sinusoidal cells in the supernatant were recovered by centrifugation for 10 minutes at 300 x g. The cells were resuspended in the presence of 28.7% Nycodenz stock solution. The final concentration of Nycodenz at this stage was 11.5%. Following centrifugation for 17 minutes at 1400 x g, The cells at the top of this Nycodenz solution were collected. Cells were resuspended in Dulbecco's modified Eagle medium supplemented with 10% fetal calf serum, The cells were seeded in 50 ml culture flask at a density of 500,000 cells/ml, The cell viability was determined by trypan blue exclusion staining, the purity of hepatic stellate cells was identified by the expression of Desmin using immunocytochemistry method. Endogenous peroxidase staining was used to detect Kupffer cells.

RESULTSThe yield rate of hepatic stellate cells was 3 x 10(7) per rat, the cell viability was more than 95%, the desmin positive cell rate was 90%, no endogenous peroxidase positive cells were detected.

CONCLUSIONThe method for the isolation of hepatic stellate cells was developed without Kupffer cells confusion. The availability of highly purified stellate cells will facilitate the investigation of their functions in primary culture.

Animals ; Cell Culture Techniques ; Cell Separation ; methods ; Kupffer Cells ; cytology ; Liver ; cytology ; Male ; Rats ; Rats, Sprague-Dawley ; Ultracentrifugation ; methods

OBJECTIVETo investigate the viability of tissue-engineered heart valve leaflets prepared with cell-polymer constructs in nude mice.

METHODSSheep endothelial cells and smooth muscle cells/fibroblasts were seeded on patches of PHA and implanted subcutaneously in athymic mice (BALB/C). The cell-polymer constructs were harvested 12, 14, 21 and 28 days after implantation.

RESULTSFourteen days after implantation, the cell-polymer constructs exhibited similar color with the autologous tissues, and HE staining showed more numerous cells in the implant. At 28 days following implantation, muscular fibers were formed in the cell-polymer constructs. V-G staining showed positive collagen staining in the implant at 12 days after implantation, while the control implants retrieved 28 days after implantation did not show extensive tissue formation or muscular fiber formation.

CONCLUSIONThe cell-polymer constructs can survive in vivo and has the potential to grow into autologous valve leaflets in the nude mice.

Animals ; Bioprosthesis ; Endothelium, Vascular ; cytology ; Heart Valves ; Implants, Experimental ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Muscle, Smooth, Vascular ; cytology ; Sheep ; Tissue Engineering ; methods

In order to study the stability of akebia saponin D (ASD) in biological fluids in vitro, the determination methods of ASD were established in this study. Akebia saponin D was dissolved in artificial gastric juice, intestinal juice and gastrointestinal contents of rats, respectively, then thermostatically maintained at 37 degrees C. At time intervals after degradation, samples were withdrawn and the concentrations of ASD were determined by HPLC, from which stability of it at different biological specimen was evaluated. As a result, ASD was totally degraded in large intestinal contents of rats in 8 hours. ASD was very stable in artificial gastric juice, intestinal juice and gastric contents of rats. All of the above data proved that ASD was easily degraded by coliform bacteria but stable in acid environment and with the presence of digestive enzyme.
Animals ; Drug Stability ; Drugs, Chinese Herbal ; administration & dosage ; pharmacokinetics ; Gastrointestinal Tract ; chemistry ; metabolism ; Humans ; Rats ; Saponins ; administration & dosage ; chemistry ; pharmacokinetics

Objective To up-regulate the expression of Glil gene in periodontal ligament stem cells ( PDLSCs) and to explore the effect of Glil gene on PDLSCs proliferation and osteogenesis differentiation by establishing Glil gene adenovirus vectors. Methods Subcloned Glil to viral backbone vector Adtrack-CMV and transfered the established vector to 293T cells, which was to acquire the virus particles. Trans-fected aim cells,namely PDLSCs,with these virus. Detected its effect on PDLSCs proliferation with CCK-8 assay, and detected the expression of Glil and the bone-related markers ALP and Runx2 through Western blot. Results An adenovirus vector, which were over expressed Glil gene, was successfully constructed and transfected to PDLSCs. Compared with the empty vector group and normal group, the over expressed one had a much slower proliferation rate in CCK-8 assay (P=0. 003). Western blot showed that ALP and Runx2 can be overexpressing os-teogenic differentiated after PDLSCs successfully transfected with the Glil gene. Conclusion Over expressing Glil gene would lead to a much slower proliferation rate in the PDLSCs and an increase of the bone-related markers. It is concluded that Glil can enhance the osteogenic dif-ferentiation capacity in PDLSCs.

Objective To study changes in serum homocysteine(Hcy) and its correlation with serum levels of folic acid and high sensitivity C-reactive protein(hs-CRP)in Tibetan patients with mildto-moderate Alzheimer's disease at various high altitude areas,so as to direct the clinical diagnosis and treatment of AD in plateau hypoxia environment Method 108 cases were divided into four groups:23 AD Tibetan patients at middle altitude(AD/middle altitude group)and 23 healthy Tibetan subjects (healthy/middle altitude group) in Qinghai-Tibet Plateau Xining region,altitude at 2,260 m,31 AD Tibetan patients (AD/high altitude group)and 31 healthy Tibetan elderly subjects (healthy/high altitude group)in Yushu region at altitude of 3,800 m.Among the total study subjects,half are males,aged from 60 to 85 years.The levels of serum Hcy,Vitamin B12 and folic acid(FA)were measured by the Fluorescence Polarization Immunoassay(FPIA).Serum hs-CRP,triglyceride(TG)and low density lipoprotein cholesterol(LDL-C)were measured by automatic biochemistry analyzer.Correlation of Hcy with FA and hs-CRP was analyzed.Result Both high altitude and middle altitude group showed the levels of Hcy and hs-CRP were significantly higher in AD Tibetan patients than in healthy control at the same altitude(allP＜0.05).The levels of Hcy,LDL-C and hs-CRP of subjects were higherat high altitude than at middle altitude(P＜0.05).In contrast,folic acid levels in AD and control groups were lower at the high altitude than at middle altitude(P＜0.05).The levels of vitamin B12 and TG were not significantly different among all four groups.Multiple logistic regression analysis showed that altitude,folacin and hs-CRP were the risk factors for Hcy in patients with AD at plateau(OR =0.351,2.794,3.021,P=0.045,0.037,0.016).Conclusion Along with increased altitude,serum level of Hcy is significantly increased in AD Tibetan patients living in high altitude area.High altitude,high hs-CRP and lower folacin may be the risk factors for hyper-homocysteine in AD Tibetan patients with high altitude,and their combined effects are involved in the occurrence and development of AD.

Objective To observe the long-term toxicity of Jiuxin Fumai Injection and to investigate the safety of clinical medication.Methods The rats were given intramuscular injection with Jiuxin Fumai Injection in large,medium,small dosage(respectively 20,10,5g? kg-1)every day for two weeks,and normal saline group served as the normal control.Two weeks after drug withdrawal,the toxic reaction in rats was observed.Results After two-week continuous administration,all the animals were alive.Some animals were vomiting and getting excited when administered the large dosage and medium dosage injection.The blood sugar elevated,the thoracic gland coefficient lowered,the hepatic cells were cloudily swollen in the animals of large dosage group.Two weeks after drug withdrawal,the above phenomenon vanished.There was no obvious toxic reaction in the small-dosage injection group.Conclusion Long-term administration of Jiuxin Fumai Injection in large dosage shows certain toxic reaction in rats.The safe dosage for intramuscular administration is less than 5 g? kg-1? d-1.

Objective To evaluate the consistency of the test results detected by multiple automated hematology analyzers .Meth-ods The EP9-A2 guideline of NCCLS ,Passing-Bablok regression analysis and t test were adopted to compare the detection results of hemoglobin(HGB) ,hematocrit (HCT) ,red blood cell count(RBC) ,white blood cell count (WBC) and platelet(PLT) in 115 samples from patients by 5 automated haematology analyzers of the Sysmex XE-2100 ,the Sysmex XT-1800i ,the Sysmex XS-1000i , the Mindray BC-5300 and ABX pentra80 .The Sysmex XE-2100 was used as the reference instrument .Results In the detection of all the compared items including HGB ,HCT ,RBC ,WBC and PLT ,these automated hematology analyzers exhibited very good cor-relation(r>0 .97) ,whereas the individual parameters in 3 automated hematology analyzers appeared the proportional systematic differences and /or constant systematic differences ,including HCT in the sysmex XT-1800i ,HCT and RBC in the sysmex XS-1000i and RBC in the ABX pentra 80 .Conclusion The Passing-Bablok regression analysis combined with t test may identify the system-atic differences of the comparative results of 2 automated haematology analyzers ,the consistency of the comparative results of the automated hematology analyzers can not be evaluated by the simple correlation coefficient .