BACKGROUND: "Toxic impaired brain retinervus" is the important mechanism of acute cerebrovascular diseases, mainly presented by cerebral microvascular impairments.OBJECTIVE: To establish cell model of various degrees ischemia-reperfusion injury and expos to qingkailing medication, in order to observe the protective role of qingkailing injection on human umbilical venous endothelium ECV304 (huveECV304)DESIGN: Randomized controlled study based on cultured cells.SETTING: Department of Combination of Traditional Chinese Medicine and Western Medicine, Union Hospital Affiliated to Tongji Medical College, Huazhong University of Science and Technology.MATERIALS: This study was carried out at Center Laboratory of Dongzhimeu Traditional Chinese Medicine College, Beijing from February to April 2002. huveECV304 were randomly divided into 7 groups, namely normal group (NG), ischemic-ypoxia group (IR group), ischemia 6 hours reperfusion 12 hours control group (I-6-hour-R-12-hour group), ischemia 24 hours reperfusion 12 hours control group (I-24-hour-R-12-hour group),ishcmeia 24 hours reperfuiosn 12 hours low dosage qingkailing group (I24-hour-R-12-hour-LDQ group), ischemia 24 hours reperfusion 12 hours middle dosage qingkailing group (I-24-hour-R-12-hour-MDQ group), ischemia 24 hours reperfusion 12 hours high dosage qingkailing group (I-24hour-R-12-hour-HDQ group) and ischemia 6 hours reperfusion 12 hours middle dosage qingkailing group (I-6-hour-R-12-hour-MDQ group). The qingkailing injection mainly consists of cholaic acid, hyodeoxycholic acid,baikal skullcap root, cape jasmine fruit and so on.were collected, and primary culture medium were removed in I-6-hour-R-12hour group and I-24-hour-R-12-hour group and added with non-glucose Earle's solution, the cells in I-6-hour-R-12-hour group were cultured in ischemic incubator in 37 ℃ for 6 hours, for 24 hours in I-24-hour-R-12-hour group and then reperfusion-cultured with Earle's medium replaced by nonLDQ group, I-24-hour-R-12-hour-MDQ group and I-24-hour-R-12-hour-HDQ group, IR injured cells were exposed to qingkailing in dosage of 5, 10,by using MTT colorimetric assay, lactate dehydrogenase leaking rate was determined by using lactate dehydrogenase reagent kit, cell apoptosis rate was detected with FCM and statistically analyzed with Cellquest software.apoptosis rate.were found gathering closely, well adhering to the wall and completely stretching, cell surface was smooth and more cleavage cells were observed,cell grew over all the bottom displaying typical "slabstone signs"; but no obvious cell morphological changes were observed in control group and Ⅰ24-hour-R-12-hour LDQ group; while in I-24-hour-R-12-hour control group, cell were found scarcely distributed and mostly drop off, a great deal of tumid round injured cells and broken cell pieces were observed,adhering cells were shrunk obviously with pseudopod stretching out, less cleavage cells were observed; In I-24-hour-R-12-hour MDQ group, cells were found well adhering to the wall, the surface was smooth without cell: Comparing to I-24-hour-R-12-hour control group, the survival rate were markedly increased in I-24-hour-R-12-hour LDQ group, I-24-hour-R12-hour MDQ group [(42.5±2.8), (72.9±8.0), (80.3±12.7)%, P ＜ 0.01], but obviously descended to (10.1 ±0.4)% (P ＜ 0.01) in I-24-hour-R-12-hour ing rate of ECV304 cell: Comparing to I-24-hour-R-12-hour control group,it was found markedly increased in I-24-hour-R-12-hour LDQ group and Ⅰ24-hour-R-12-hour MDQ group [(6.6±1.4), (2.4±1.4), (2.4±0.5)%, P ＜ 0.01], but significantly ascended to (16.1±2.2)% in I-24-hour-R-12-hour rate of ECV304 cell: Comparing to I-6-hour-R-12-hour control group, it was obviously increased in comparing to I-6-hour-R-12-hour MDQ group [(2.2±0.5), (1.5±0.4)%, P ＜ 0.05]; comparing to normal group, it was found significantly increased in I-24-hour-R-12-hour control group and I-24-hourR-12-hour MDQ group [(0.9±0.2), (5.5±2.5), (9.9±4.3)%, P ＜ 0.01]; Moreover the increment in I-24-hour-R-12-hour control group was significantly higher than I-24-hour-R-12-hour MDQ group (P ＜ 0.01).ECV304 with various degree IR injury, but which displaying tow opposite cells due to I-6-hour-R-12-hour exposure was mainly presented by lowered cell apoptosis rate; But severer injury of ECV304 cells due to I-24-hour-R12-hour exposure was presented by increased cell apoptosis rate and lowon and cell membrane would not break in apoptostic cells and no internal content effused, therefore the surrounding cells would not be affected,which increasing the survival rate and displaying protective role for most cells.

Animals ; China ; epidemiology ; Dogs ; Humans ; Rabies ; epidemiology ; prevention & control ; Rabies Vaccines

Vascular endothelial growth factor (VEGF) have multiple protective effect on the nervous system which related to angiogenesis,neurogenesis,directly neurotrophic protection and antiapoptotic mechanisms.The study of the nervous system of VEGF focus on tumors,trauma,stroke,autoimmune diseases and neurodegenerative diseases.The study of VEGF related to epileptic seizures is relatively rare.This paper is to make a review about the neuroprotective effect of VEGF on epileptic seizure.

Objective: To investigate the effects of neferine ( Nef ) on the learning and memory function and the expression of Notch1 and SYN in hippocampus in the rats with chronic cerebral ischemia .Methods:Male SD rats (250-300g) were randomly divid-ed into the sham operation group(Sham), the model group(Mod), nimodipine (Nim) positive control group and Nef treatment group with 5 ones in each .The chronic cerebral ischemia ( CCI ) model was established using bilateral common carotid artery ligation ( 2-VO).The rats were orally administered with NS(5 mg· kg-1),Nim(1 mg· kg -1) and Nef (20 mg· kg-1),respectively for 21 days after the first day of operation .The body weight was recorded .The number of hippocampal neurons in the rats was observed by Nissel staining.The learning and memory function was evaluated by Morris water maze test .Notch1 and SYN protein expressions in hippo-campus were detected by IHC staining and Western blot .Results:Compared with the Mod group , Nef group could reverse the reduc-tion of body weight and the number of hippocampal neurons in hippocampus CA 1 region induced by 2-VO, increase the route and the time of platform finding, prolong the escape latency and decrease the number of platform cross on the 21st day, and the effects were bet-ter than those of Nim (P<0.05).Moreover, the down-regulated SYN and Notch1 protein expressions in CCI group were both increase after the Nef treatment on the 21st day.Conclusion:Nef has a protective effect on the function of learning and memory in CCI rats in-duced by 2-VO,which may be related with the strengthened Notch 1 pathway in CA1 region of hippocampus and synaptic plasticity .

There are many kinds of scaffolds used in the field of tissue engineering, which can be divided into natural biological scaffolds and synthetic scaffolds. This paper summarizes the latest advances of research on scaffolds of skin tissue engineering and review amounts of related literatures on this field. The result shows that natural biological scaffolds can provide a better three-dimensional space for the adhesion and generation of cultured cells. These scaffolds have an extensive application in skin tissue engineering.

OBJECTIVETo observe the clinical efficacy difference in the treatment of neonatal hypoxic ischemic encephalopathy (NHIE) between the combined therapy of acupuncture land acupoint injection of nerve growth fac-tor (NGF) and the combined therapy of the intravenous drop of citicoline sodium and intramuscular injection of NGF.

METHODSSixty-one cases of NHIE were randomized into an observation group (32 cases) and a control group (29 cases). In the observation group, acupuncture combined with acupint injection of NGF was adopted. The acupoints were Baihui (GV 20), Dazhui (GV 14) and Fengfu (GV 16), 0. 5 mL each acupoint, once a day. In the control group, the intravenous drop of citicoline sodium and intramuscular injection of NGF were used. The, treatment of 7 days made one session and 3 sessions were required. Separately, in 7, 14 and 21 days of treatment, the clinical efficacy, behavior nerve determination score and adverse reaction were observed in the two groups.

RESULTSIn 7, 14 and 21 days of treatment, the total effective rates were 50. 0% (16/32), 68. 8% (22/32) and 78. 1% (25/32) in the observation group and were 69. 0% (20/29), 72. 4% (21/29) and 82. 8% (24/29) in the control group. The result in the control group was better than that in the observation group in the 7 days (P< 0. 05). The behavior determination scores in 7, 14 and 21 days were all improved obviously as compared with those before treatment in the two groups (P<0. 05, P<0. 01). The score in the control group was better than that in the, observation group in the 7 days (P<0. 05). The differences were not significant in the scores between the two groups in 14 and 21 days (both P>0. 05). The incidence of adverse reaction in the observation group was lower than that in, the control group [14. 3% (5/35) vs 31. 4% (11/35), P<0. 05] and the severity was mild.

CONCLUSIONIn the treatment of NHIE with the combined therapy of acupuncture and acupint injection, the overall efficacy in 1 week; is inferior to that of the combined therapy of the intravenous drop of citicoline sodium and intramuscular injection of NGF. But the efficacy is improved gradually along with the treatment progression and the adverse reaction is less.

Acupuncture Points ; Acupuncture Therapy ; Female ; Follow-Up Studies ; Humans ; Hypoxia-Ischemia, Brain ; drug therapy ; therapy ; Infant ; Infant, Newborn ; Male ; Nerve Growth Factor ; administration & dosage

Animal Diseases ; epidemiology ; prevention & control ; virology ; Animals ; China ; epidemiology ; Humans ; Infection Control ; methods ; Rabies ; epidemiology ; prevention & control ; veterinary ; virology ; Rabies Vaccines ; administration & dosage ; Rabies virus ; immunology

Genetic Engineering ; Humans ; Rabies ; virology ; Rabies virus ; genetics ; pathogenicity ; Viral Proteins ; genetics ; Virulence

Objective To discuss the possibility of constructing a new kind of active skin substi tutes.Methods The culture of dermal fibroblasts were isolated from foreskin of an infant; preparation of human acellular amniotic extracellular matrix was conducted through the disposition of detergent and enzyme.Surface structure porosity and pore size were detected through scanning electron microscopy.bFGF-chitosan gelatin microspheres were prepared.The features such as the size distribution,drug content,drug encapsulating and in vitro release were studied with scanning electron microscopy,laser grainsize analyzer and ELISA method.MTT test was performed to observe cell proliferation and evaluate the biocompatibility in vitro.Results The cellular layer of amniotic membrane was completely removed but did not damage the collogen scaffolds structure by the disposal of enzyme and detergent.Scanning electron microscopy showed that the scaffolds had criss-cross structure and high porosity,the pore size was irregular and varied from 10 nm to 100 nm.Scanning electron microscopy of bFGF- gelatin-chitosan microspheres surface structure showed that the microspheres had spherical structure,uniform size and smooth surface quality.Controlled release curve showed that the sudden release of bFGF was obvious.Conclusions A new type of active skin substitutes is prepared through culturing fibroblasts on HAAM loaded controlled-released basic fibroblast growth factor from chi tosan-gelatin microspheres.

AIM:To investigate the effects of fucoidan on the angiogenesis of multiple myeloma cells in vitro, and its related mechanisms .METHODS:The human multiple myeloma RPMI 8226 cells and human endothelial cells were cultured in vitro.The growth inhibition rate of RPMI 8226 cells was examined by MTT assay .The cell cycle and apoptosis rate were measured by flow cytometry .RPMI 8226 cells were treated with fucoidan for 72 h, and the cell culture superna-tant was collected .The VEGF concentration was examined by ELISA , and the tube formation assay was applied to assess the angiogenic activity .After treatment with fucoidan for 72 h at different concentrations , the protein levels of HIF-1α, VEGF, p-AKT and p-ERK1/2 were detected by Western blot .RESULTS: Fucoidan inhibited the growth of RPMI 8226 cells in a dose-and time-dependent manner .After treatment with fucoidan for 72 h, the cell cycle was arrested at G 1 phase , and the apoptotic rate of RPMI 8226 cells was increased with the increasing concentration of fucoidan , which was much higher than that in control group (P<0.05).The VEGF concentration was significantly decreased with the increa-sing concentration of fucoidan .The numbers and areas of the capillary-like structures decreased while the concentration of fucoidan increased, and those at 100 mg/L were less than those in the control (P<0.05).The protein levels of HIF-1α, VEGF, p-AKT and p-ERK1/2 in fucoidan group were significantly lower than those in control group (P<0.05).CON-CLUSION:Fucoidan inhibits the secretion of VEGF in multiple myeloma cells , and reduces angiogenesis induced by mul-tiple myeloma cells .It inhibits the protein expression of HIF-1αand VEGF , which may be related to inhibiting the phos-phorylation of AKT and ERK1/2.