BackgroundEndostatin (ES) is currently the strongest endogenous angiognesis inhibitor,and it can inhibit the occurrence of neovascularization.Various studies demonstrated that the poly RGD sequence can enhance the function of the ES gene.ObjectiveThis study was to evaluate the use of gene therapy of modified ES for alkaline burn-induced corneal neovascularization (CNV).MethodsOne hundred and two clean SD rats were randomly divided into the normal control group,the pCI empty vector group,the pCI-ES group,and the pCI-RGDRGDES group.Corneal neovascularization models were established by placing a piece of 3 mm filter paper with 1 mol/L NaOH at the central cornea for 40 seconds.3 μg of the pCI blank vector,ES-tranfected pCI blank vector,or RGDRGD-ES-transfected pCI vector was injected into the superior bulbar conjunctiva after the alkali burn twice at 1-week intervals.Area of CNV and edema of the cornea in the various groups of rats were examined daily under the slit lamp biomicroscope.1,4,7 and 14 days after operation,the rats were sacrificed by the excessive anesthesia method and corneal tissues were obtained to evaluate pathological changes.The expression of CD34 in vascular endothelial cells was detected by immunochemistry to calculate the corneal neovascular density.The expressions of VEGF mRNA and Flk-1 protein in the corneas were detected by reverse transcription polymerase chain reaction (RT-PCR) and Western blot analysis.The use and maintenance of animals followed the Statement of ARVO.Results Seven to fourteen days after corneal alkali-burning,the corneal neovascular area was smaller in the pCI-ES group and pCI-RGDRGD-ES group compared with the normal control group and pCI blank vector group (P＜0.05,P＜0.01 ),and nevascular area in the pCI-RGDRGD-ES group was smaller than that in the pCI-ES group (P＜0.05).The expression level of CD34 was significantly lower in the pCI-ES group and pCI-RGDRGD-ES group than that in the normal control group and pCI blank vector group (P＜0.05,P＜0.01 ),and the expression level of CD34 was further declined in the pCI-RGDRGD-ES group compared with the pCI-ES group (P＜0.05 ).Compared with the normal control group and pCI vector group,the expressions of the Flk-1 protein and VEGF mRNA were decreased in the pCI-ES group and pCI-RGDRGD-ES group on the fourth day after corneal alkali-burning (P＜0.01,P＜0.05 ),and those in the pCI-RGDRGD-ES group were less than the pCI-ES group (P＜ 0.05,P＜ 0.05 ).Conclusions Subconjunctival injection of both ES and modified RGDRGD-ES genes result in significant suppression of CNV in vivo,and modified RGDRGD-ES appears to be more effective than native ES.The main mechanism of ES in inhibiting neovascularization is to downregulate the expression of VEGF and Flk-1.

OBJECTIVETo assess how trace element selenium and B27 supplements affect the neural stem cell (NSc) differentiation in vitro.

METHODSThe development and differentiation of NSc from the newborn rat were observed with primary culture and subculture during treating by sodium-selenite, and selenium-methyl-cysteine (SMC). The immunocytochemistry techniques were used to identify the NSc and mature protein expression with neuron marker beta-tubulin, astrocyte marker GFAP, and oligodendrocyte marker CNPase. The neurosphere morphology and neurite outgrowth were observed.

RESULTSAdding the complete B-27 serum-free supplement, Selenium could promote the neurosphere viability, development and differentiation. Without selenium and B-27, neurosphere could not survive and differentiate. Without B-27 in the medium but there containing selenium, the neurosphere could promote the viability and development into neuron, astrocyte and oligodendrocyte, as compared with the no-containing B-27 and selenium groups, these differentiated cells might have more quantity, more branches and better morphological nerve net. The count of the neuron, astrocyte and oligodendrocyte was 11.2/Hp, 16.1/Hp and 9.3/Hp.

CONCLUSIONSThe selenium should be very important for neural stem cells' survival. Selenium could promote the neurosphere cells differentiation and development.

Animals ; Animals, Newborn ; Cell Differentiation ; drug effects ; Cell Survival ; drug effects ; Cells, Cultured ; Culture Media, Serum-Free ; pharmacology ; Cysteine ; analogs & derivatives ; pharmacology ; Female ; Glial Fibrillary Acidic Protein ; metabolism ; Immunohistochemistry ; Male ; Neurons ; cytology ; drug effects ; metabolism ; Organoselenium Compounds ; pharmacology ; Rats ; Rats, Wistar ; Selenium ; pharmacology ; Selenocysteine ; analogs & derivatives ; Sodium Selenite ; pharmacology ; Stem Cells ; cytology ; drug effects ; metabolism ; Tubulin ; metabolism

Background Corneal epithelial abrasion results in corneal ulcer and stroma cloudy evenb irreversible visual impairment.Previous drugs for corneal epithelial injury can only alleviate the inflammatory irritation.So it is very important to seek a drug which regulate the growth of corneal epithelium.Objective This study was to investigate the effects of recombinant human BIGH3 protein eye drops on corneal epithelial abrasion.Methods Fifty right eyes of 50 clean adult New Zealand white rabbits were collected.Two rabbits were sacrificed right away following establishment of corneal epithelial abrasion models (0 hour group).The other 48 rabbits were randomly divided into recombinant human epidermal growth factor (EGF) derivative group (positive control group),normal saline solution group (negative control group),0.25％ or 0.5％ recombinant human BIGH3 protein eye drops group.Corneal abrasion models were created with alcohol corrosion method with a defect area of 7 mm2.The corresponding eye drops were used separately in 4 groups for four times per day after operation.Experimental eyes were examined by the slit lamp microscope,and fluorescein vital staining were performed 12,24,36,48,72 hours after operation.Planimetry was performed and the corneal photographs were analyzed with computer software.The rabbits were sacrificed 12,24,36,48 and 72 hours after operation,respectively,and the histopathological examination of corneal tissue was carried out.Results No obvious irritation response was seen after administered of eye drops in the recombinant human EGF derivative group,normal saline solution group,0.25％ and 0.5％ recombinant human BIGH3 protein eye drops groups.Histopathological examination revealed a full-thickness defect of corneal epithelium after modeling.The defect area was gradually smaller with time lapse,and corneal epithelium migrated from periphery toward the center zone.Corneal epithelial cells increased with time lapse.Compared with normal saline solution group,the defect area of corneal epithelium lessened 12,24,36,48 hours after operation in the 0.25％,0.5％ recombinant human BIGH3 protein eye drops groups and recombinant human EGF derivative group (all at P =0.000),but at 12and 24,36 hours after operation,no significant differences were found between the recombinant human EGF derivative group and normal saline solution group (P =0.321,0.057,0.126).The defect area was smaller in the 0.5％recombinant human BIGH3 protein eye drops group than that of the recombinant human EGF derivative group at various time points (P=0.042,0.039,0.025,0.008).However,significant smaller defect area was exhibited only at 12 hours and 24 hours after operation in the 0.25％ recombinant human BIGH3 protein eye drops group (P=0.047,0.042).No significant differences were seen in corneal defect area at various time points between 0.25％ and 0.5％recombinant human BIGH3 protein eye drops groups (P =0.358,0.259,0.108,0.062).In addition,the corneal defect area was (0.51 ±0.42)mm2 72 hours after operation in the normal saline group;while that in the recombinant human EGF derivative group and recombinant human BIGH3 protein eye drops groups was disappeared.The repairing curves in the recombinant human BIGH3 protein eye drops groups were superior to those of the recombinant human EGF derivative group and normal saline solution group.Conclusions 0.25％ and 0.5％ recombinant human BIGH3 protein eye drops have facilitation effect on the growth of corneal epithelial cells and the healing of corneal injury.

OBJECTIVETo analyze the outbreak patterns of acute schistosomiasis in Anhui province in 2003.

METHODSThe criteria of emergency event for schistosomiasis issued by the Ministry of Health were applled in this study. Epidemiological data of acute schistosomiasis outbreak in Anhui province in 2003 were collected, checked and analyzed.

RESULTSIn 2003, there were 6 episodes of acute schistosomiasis outbreaks in Anhui which had met the criteria of being emergency events, issued by the Ministry of Health. The preplanning was initiated in 5 outbreaks to respond to the outbreaks of acute schistosomiasis. All of the acute schistosomiasis cases received timing treatment, except those misdiagnosed cases coming from the non-endemic areas. There were no deaths or follow-up cases during the outbreaks. Health education, preventive treatment for people living in high risk villages were conducted in order to timely control the epidemics of schistosomiasis.

CONCLUSIONThe implementation of preplanning has played an important role in early cases-finding, timing report and response to the outbreaks of acute schistosomiasis.

Adolescent ; Adult ; Animals ; Cattle ; Child ; Child, Preschool ; China ; epidemiology ; Disease Outbreaks ; Disease Reservoirs ; Female ; Health Education ; Humans ; Male ; Middle Aged ; Prevalence ; Schistosomiasis japonica ; drug therapy ; epidemiology ; prevention & control ; Snails ; parasitology

OBJECTIVETo construct a replication-incompetent adenovirus vector targeting cancer stem cells by modified touchdown PCR and overlap extension PCR and investigate its infection efficiency in CD133(+) SW480 cells in vitro.

METHODSThe two portions of the fiber gene encoding the Ad5 fiber knob domain with the HI loop deleted were amplified using two pairs of designed primers and then linked by overlap extension PCR. The product obtained was identified by sequencing and inserted into prokaryotic expression vector pEGFP-N1. The product, pEGFP-N1 KNOBδHI, contained a unique EcoRV restriction site in the deleted portion of the sequence encoding the HI loop. The gene sequences of the adenovirus fiber were amplified using both common PCR and overlap extension PCR, then identified by sequencing and inserted into pNEB193, resulting in pNEB-F5. CD133(+) SW480 cells were infected with the generated adenovirus vectors Ad5-GFP and Ad5FHI-GFP to investigate the infection efficiency using fluorescent microscope.

RESULTSThe target fragments of expected sizes were amplified by touchdown PCR and overlap extension PCR, but not by common PCR. Ad5FHI-GFP showed a higher infection efficiency than Ad5-GFP in CD133(+) SW480 cells.

CONCLUSIONCompared with common PCR, touchdown PCR and overlap extension PCR can significantly improve the specificity and efficiency of the PCR products for constructing CD133(+) cancer stem cell-selective adenovirus type 5 vector, which provides carriers for tumor-targeted gene therapy.

AC133 Antigen ; Adenoviridae ; genetics ; Antigens, CD ; Cell Line, Tumor ; Genetic Vectors ; Glycoproteins ; Humans ; Neoplastic Stem Cells ; cytology ; virology ; Peptides ; Plasmids ; genetics ; Polymerase Chain Reaction ; methods

OBJECTIVETo probe rational indices suitable for ecological surveillance on breeding ground for Oncomelania hupensis snails in areas prevalent with islet-type schisitosomiasis using remote sensing technology.

METHODSThree adjacent islets, prevalent with islet-type schistosomiasis, along the Yangtze River within the boundaries of Dongzhi County, Anhui Province were selected as study field for remote sensing analysis. Multi-spectral data were composed and non-supervisedly classified in computer with Idisi software for remote sensing analysis. Values of the normalized difference vegetation index (NDVI), green vegetation index (GVI), bright index (BI), which reflect the greenness and brightness of landscape, were also calculated. Finally, all the results were comprehensively analyzed, combined with data from the field investigation.

RESULTSNDVI, GVI and BI could depict characteristics of the landscape quantitatively. Values of NDVI, BI and GVI were varied in different types of landscapes, and 95% confidence interval of these values suitable for breeding of snails was 0.0522 approximately 0.3566, 2.4162 approximately 28.2672 and 29.3404 approximately 40.3135, respectively. Classification of NDVI showed that type 5 anf type 6 were main breeding ground for snails, and type 4 with values of NDVI from 0 to 0.1 was potential areas for snail propagation. Classification of GVI showed that types 5, 6 and 7 were main breeding ground for snails, and also type 4 with values of GVI from 2 to 10 was potential areas for snail propagation. Both NDVI and GVI showed type 2 and type 3 were temporarily not suitable for snail breeding.

CONCLUSIONIndex figures of NDVI and GVI re-formed by reasonable classification could reflect not only breeding ground for snails and range of the areas for snail propagation in islets, but also their evolving rules, i.e., status of new marshland formation and vegetation growth.

Animals ; Breeding ; Ecology ; Satellite Communications ; Schistosomiasis ; transmission ; Snails ; parasitology ; physiology

OBJECTIVETo compare the effect of the extracts from Decoction for resuscitation (DRE) and its component herbs on prostacyclin (PGI2), thromboxane A2 (TXA2) and nitric oxide (NO) release from rat vascular endothelial cells under hypoxia.

METHODAfter treatment with the extracts from DRE and its component herbs, the contents of 6-keto-prostaglandin F1alpha(6-keto-PGF1alpha), thromboxane B2 (TXB2) as well as nitrite (NO), which were degradation products of PGI2, TXA2 and NO respectively, in culture medium of rat vascular endothelial cells under hypoxia were measured with radioimmunoassay and Griess Reaction.

RESULTCompared with the control group, the results indicated that DRE, prepared licorice root extract (LE), dried ginger extract (GE), aconite root extract (AE), extracts of aconite root and prepared licorice root (ALE), extracts of aconite root and dried ginger (AGE) increased significantly the content of 6-keto-PGF1alpha and the ratio of 6-keto-PGF1alpha/TXB2, but had no effect on the content of TXB2 in culture medium of rat vascular endothelial cells under hypoxia. The content of 6-keto-PGF1alpha in the DRE group was higher than that in the groups of LE, GE, AE, ALE, AGE. The ratio of 6-keto-PGF1alpha/TXB2 in the DRE group was higher than that of the groups of GE, AE, ALE. Compared with the control group, DRE, LE, GE, AE, ALE, AGE increased significantly the content of NO2- in culture medium of rat vascular endothelial cells under hypoxia. Moreover, the content of NO2- in the DRE group was higher than that of the groups of GE, AE, ALE.

CONCLUSIONThe results suggested that DRE increased significantly the content of PGI2 and the ratio of PGI2/TXA2 as well as the content of NO. The effect of DRE on the parameters in culture medium of rat vascular endothelial cells under hypoxia was better than that of the extracts from its component herbs.

6-Ketoprostaglandin F1 alpha ; metabolism ; Aconitum ; chemistry ; Animals ; Aorta, Abdominal ; cytology ; Cell Hypoxia ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Endothelial Cells ; metabolism ; Ginger ; chemistry ; Glycyrrhiza uralensis ; chemistry ; Nitric Oxide ; metabolism ; Plants, Medicinal ; chemistry ; Rats ; Rats, Wistar ; Thromboxane B2 ; metabolism

AIMTo investigate the correlation between the gastric adaptive cytoprotection and the low concentration alcohol intake in a chronic drinking rat model and the effect of chronic ethanol exposures on the cell turnover of the gastric mucosa and its possible role in adaptive cytoprotection.

METHODSSprague-Dawley rats received the drinking water containing 6% (v/v) ethanol as their only water intake for 28 days. In the different stages of the 28 days (1st, 3rd, 7th, 14th and 28th days), the stomachs of the rats were cannulated and perfused with pure ethanol, and the severity of mucosal lesions was measured in 2 hours at the end of perfusion respectively. The cell proliferation and apoptosis in gastric mucosa of rats in different groups were analyzed by flow cytometer, immunohistochemistry and computer image analysis.

RESULTSPure ethanol caused ulcer and haemorrhagic damage in the corpus and antral mucosa of the control rats. These lesions were prevented by pretreatment of the animals with ethanol exposure in the 3 rd to 14 th days. The damage index was decreased by 80%, as compared with those in control rats. There was no significant difference in the rats exposed to the ethanol in the 1st and 28th days. Compared with control, the cell apoptosis in gastric mucosa of the rats was enhanced during they exposure to the ethanol in the 3rd to 28th days. Otherwise the cell proliferation was increased in the 3rd to 28th days, and decreased in the 28th days, respectively.

CONCLUSIONChronic adequate alcohol intake may enhance the cell turnover of gastric mucosa and lead to an adaptive cytoprotection. Long-term stimulus with the low concentration ethanol may cause the atrophy of gastric mucosa and reduce the gastric mucosal cytoprotective effect.

Alcoholism ; Animals ; Apoptosis ; Cell Proliferation ; Cytoprotection ; Ethanol ; adverse effects ; Gastric Mucosa ; cytology ; pathology ; Male ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo improve the quality of medical instrument clinical trial in China.

METHODSA systematic analysis on current situation of medical instrument clinical trial in China is conducted to find a way out of problems.

RESULTS & CONCLUSIONSThere is a low level of medical instrument clinical trial now in China. There are still many shortcomings in law and regulations of medical instrument clinical trial. The sponsors and s do not know medical instrument clinical trial well, the measures of quality related to medical instrument clinical trial must be improved.

China ; Clinical Trials as Topic ; standards ; Equipment and Supplies ; Humans

OBJECTIVE: To observe the inductive efficiency of deriving hematopoietic cells from human embryonic stem (hES) cells co-cultured with human yolk sac stromal cells, fetal liver stromal cells or fetal bone marrow stromal cells,in order to discuss the effect of the different hemopoietic microenvironment on hemopoietic cytogenesis. METHODS: We used two-step method to induce the hES cells into the hematopoietic cells. In the first step the hES cells were co-cultured with cytokines by formation of the day 5 embryoid bodies (5d EBs). In the second step the 5d EB cells were induced into the hematopoietic cells by co-culturing with human yolk sac stromal cells, fetal liver stromal cells or fetal bone marrow stromal cells for 10 days. The inductive efficiencies of deriving hematopoietic cells from hES cells co-cultured with the different hemopoietic microenvironment were reflected by the expression levels of flk, CD34 and CD45 antigen. RESULTS: Flow cytometry analysis demonstrated that the population of the cells co-cultured with human yolk sac stromal cells contained flk (1.80%+/-0.56%), CD34 (1.30%+/-0.14%) or CD45 (1.05%+/-0.63%) positive cells; the population of the cells co-cultured with human fetal liver stromal cells contained flk (34.00%+/-25.45%), CD34 (38.40%+/-24.80%) or CD45 (72.60%+/-25.70%) positive cells; the population of the cells co-cultured with human fetal bone marrow stromal cells contained flk (2.50%+/-1.48%), CD34 (3.20%+/-0.56%) or CD45 (1.65%+/-0.21%) positive cells. Compared with spontaneous differentiation of EBs, all of the three stromal cells could induce EBs into the hematopoietic cells (P<0.05). CONCLUSION The inductive efficiency of deriving hematopoietic cells from EBs co-cultured with human fetal liver stromal cells was higher than EBs co-cultured with human yolk sac stromal cells and fetal bone marrow stromal cells.
Antigens, CD34 ; Cell Differentiation ; Cells, Cultured ; Cellular Microenvironment ; Coculture Techniques ; Embryonic Stem Cells ; cytology ; Fetus ; cytology ; Hematopoietic Stem Cells ; cytology ; Humans ; Leukocyte Common Antigens ; Mesenchymal Stem Cells ; cytology ; Stromal Cells ; cytology ; Yolk Sac ; cytology