Objective To culture astrocytes of human optic nerve and establish the cell lines for further study of healing process after optic nerve trauma. [WT5”HZ]Methods [WT5”BZ]Astrocytes of infantile optic nerve were cultured by tissue inoculation or tissue digestion with 0.25% trypsin and 0.06% EDTA. The second and fourth passage cells were stained with HE and anti GFAP, S 100 protein, vimentin, and CD34 antibodies. [WT5”HZ]Results The trypsinized astrocytes of infantile optic nerve reached confluence in 7 days. The cultured cells were in polygonal shape with processes and the cytoplasm was abundant. These cells were positive in GFAP, S 100 protein and vimentin staining, and negative in CD34 staining. Conclusions Astrocytes of human optic nerve can be successfully cultured by trypsinization rather than tissue inoculation.

To explore the relationship between serum resistin concentration, hypertension, degree of obesity and blood glucose, fasting serum resistin concentration was measured with EIA in 17 normotensives and 44 patients with essential hypertension. Serum resistin concentration was significantly correlated with age and body fat percectage (BF%). The concentration level was significantly higher in grade 2 hypertension group [(30.5?11.0)?g/L] than in grade 1 hypertension group [(24.3?10.5)?g/L] and in normotensives group [(21.5?6.1)?g/L]. The difference in serum resistin concentration between the three groups disappeared when age and BF% were corrected by analysis of covariance. Serum resistin concentration was significantly correlated with systolic blood pressure, age, BF%, fasting plasma glucose. A stepwise multiple linear regression analysis revealed that age, BF% and fasting plasma glucose were significantly independent predictors of serum resistin concentration. The results suggest that serum resistin concentration is not directly related to blood pressure, but the concentration level is actually correlated with age, BF% and glucose metabolism.

Objective To investigate the effect of obesity on change of the levels of blood glucose and blood lipid after treatment with different combinations of antihypertensive drugs in patients with essential hypertension (EH). Methods One hundred and thirty-eight patients with mild to moderate EH were randomly divided into 3 groups with different combinations of drugs: reserpine with dihydralazine, hydrochlorothiazide, potassium chloride group (group 1), fosinopril with indapamide group (group 2), and nifedipine with atenolol group (group 3). Treatment period lasted for 14 months. Every group was divided into obesity subgroup and non-obesity subgroup according to Chinese obesity standard. Results After treatment, blood glucose levels were significantly decreased and blood lipid level was unchanged in both obesity subgroup and non-obesity subgroup of group 1. High density lipoprotein-cholesterol (HDL-C)/low density lipoprotein-cholesterol (LDL-C) ratio was significantly decreased and blood glucose was significantly increased in obesity subgroup, however, there was not significant change in the level of blood glucose and blood lipid in non-obesity subgroup of group 2. Blood glucose level was significantly increased and HDL-C/LDL-C ratio was significantly decreased in obesity subgroup, however, there was significant increase only in fasting blood glucose in non-obesity subgroup of group 3. Conclusions The results suggested that treatment with a combination of reserpine and dihydralazine, hydrochlorothiazide, potassium chloride showed beneficial effect on improving carbohydrate metabolism in both obese and non-obese patients with EH, while the treatment with the combination of fosinopril with indapamide deteriorated carbohydrate and lipid metabolism in obese patients with EH, and the treatment with combination of nifedipine with atenolol impaired carbohydrate and lipid metabolism in obese patients with EH.

To determine the effect of pre-operative chemotherapy on apoptosis in breast cancer and to evaluate its signif icance as a prcgnostic marker. MethodsPatients with breast cancer were divided into preoperative chemotherapy group (40 cases)and control group (42 cases). Two groups were analyzed for the appearance of apoptosis by using TUNEL method and electron mi croscope in tissue sections. ResultsApoptosis occurred in 92.5 % of preoperative chemotherapy group and in 78.5 % of control group. The apoptotic indexes were 19.37 + 6.49 and 9.26 + 5.04 ( P ＜ 0.01 ) respectively. Low apoptotic index was related to disease-free survival of patients with breast cancer (P ＜ 0.01 ). ConclusionThe preoperative chemotherapy can induce apoptcsis of breast cancer and improve disease-free survival.

Objective To determine the relationship between serum levels of MCAF/MCP-1 (monocyte chemotactic and activating factor/monocyte chemoattractant protein-1), RANTES(regulated on activation, normal T-cell expressed and secreted) and the disease activity of systemic lupus erythematosus(SLE). Methods Serum levels of MCAF and RANTES were measured by ELISA. Results ①Serum level of MCAF but not RANTES, was significantly increased in patients with SLE as compared with controls. ②Serum level of MCAF but not RANTES, was markedly higher in patients with active disease than those with inactive disease. ③No significant differences were found in the serum levels of MCAF and RANTES between patients with renal damage and those without. Conclusions These results suggest that MCAF may be involved in the pathogenesis of SLE, and serum MCAF levels could be an indicator for the disease activity of SLE.

OBJECTIVE:To study the effect of dexamethasone (Dex) on expression of aquaporin-1(AQP1) in cultured human trabecular meshwork(HTM) cells METHODS:Reverse transcription combined with polymerase chain reaction(RT-PCR) was used to detect the expression of AQP1 in cultured HTM cells and those treated by Dex RESULTS:The mRNA of AQP1 expressed in normal HTM cells was 1 643?0 354,while 1 577?0 405,1 117?0 443,0 458?0 301,0 267?0 243 in those treated with Dex for 3 days in concentrations of 10-8mol,5?10-8 mol,10-7 mol,5?10-7mol As the concentrations of Dex increased to≥5?10-8mol,the expression of AQP1 was inhibited(P

In order to estimate the relationship between iron and the Alzheimer's disease, the behavioral and pathological changes were observed by Morris water maze and immunohistochemical staining respectively after injecting FeC13 into brain ventricle of rats. The apoptosis was tested by flow cytometry and the electron microscopy was used to observe ultrastructural changes. There were significant differences in escape latency of time and distance between normal animals and iron treated rats. Percentage and fluorescence intension (FI) of AnnexinV FITC loaded cells undergoing apoptosis were higher in iron treated rats compared with normal animals. Fawn-coloured products of β amyloid protein were interspersedly distributed in extensive areas of cerebral cortex and hippocampus. Under electron microscope, vacuolate degeneration of neuronal processes with mitochondria degeneration and accumulation of microtubule near vacuolar nucleus were observed in iron treated rats. These results suggest that a local higher concentration of iron in brain may induce Alzheimer-like impairment of intelligence and pathological changes.

BACKGROUND: It is important to study the methods of culturing bone marrow-derived mesenchymal stem cells (MSCs) to obtain a great amount of high purity MSCs for applying ocular tissues constructed by tissue engineering technique to treat eye diseases.OBJECTTVE: To separate and culture in vitro MSCs from bone marrow of the adult rats by density gradient centrifugation combined with adherence culture, and observe the growing characteristics and the possibility of mass multiplication.DESIGN: A completely randomized grouping design/repetitive measuring experiment.SETTING: Pathological laboratory, Zhongshan Ophthalmic Center, Sun Yat-sen University.MATERIALS: Four six-week-old SD rats about 250 g, grade Ⅱ of cleaning, were provided by the Animal Center of Sun Yat-sen University [certificate number: SCSK(Yue)2004/0011], about 250 g each rat and there was no limit to the sex. The main reagents and instruments included low sugar Dulbecco modified Eagle culture medium (DMEM/F12, American Gibco Corporation), trypsin (fetal bovine serum (FBS, Hangzhou Sijiqing Bio-Engineering Material Research Institute), American Gibco Corporation), disodium edetate, lymphocyte separating medium, fibronectin, CD44, CD34, CD31 monoclonal antibodies, two-step-method kit for immunohistochemistry (Beijing Zhong shan Biotechnology Corporation).METHODS: This experiment was conducted at the Key laboratory of Ophthalmology (Sun Yat-sen University), Ministry of ethanol (750 g/L) for 10 minutes. Under aseptic condition, the medullaris cavitas was exposed, the syringe containing application m edium was directly punctured into the femoral cavity, the cells in the medullaris cavitas were washed out with the culture medium containing heparin and taken as the cell suspension. The bone marrow-derived MSCs were separated and purified by density gradient centrifugation combined with adherence culture, and the growing conditions of the wells. When the cells had generally connected with each other, they were fixed with methanol or dimethy ketone in situ for 10 minutes, and then hematoxylin eosin (HE) staining or immunohistochemical staining. Antibodies against fiinoculated to 96-well culture plate by a cell density of 4.25×107/L, with 200 μL every well; then put the culture plate into culture box. Then from the next day to the sixth day, 5 g/L MTT solution was added into two rows (20 μL every well) every day, continuously cultured for 4 hours, then the supernatant was removed, and 200 μL DMSO was added to each well, agitated for 5 minutes, then detected the absorbance (A) values at 570 nm wave length, and a growth curve was drawn.branes were clear and the cell bodies were lucent. Being cultured for 2 days, there appeared adherent cells. 3 days later,most of the adherent cells extended and appeared to be in polygon, star or long-shuttle shapes. 4 days later, the cells showed to be in division growth stage; and about 12 days later, the cell clones were connected to each other, appearing curve of subcultured MSCs was in S shape. Cells began growing fast on the 2nd day after being passaged, and they entered the growth period for 3-4 days followed by saturation period, and then cells stopped growing.CONCLUSION: It is a simple and practical method to separate MSCs from the bone marrow of adult rats by means of density gradient centrifugation and adherence. MSCs can greatly proliferate in vitro and offer seed cells for the application of tissue engineering technique to treat eye diseases.

AIM: To investigate the effects of internal change of serum insulin and plasma glucose levels on serum free fatty acid (FFA) concentrations after glucose loading. METHODS: Serum insulin, plasma glucose and FFA concentrations were measured simultaneously in 234 essential hypertension patients who were undergoing oral glucose tolerance test (OGTT) [ including 20 cases with 2 type diabetes mellitus (DM), 74 impaired glucose tolerance (IGT), 140 normal glucose tolerance (NGT); 98 males, 136 females]. RESULTS: Fasting serum FFA concentration (?mol/L) in DM ((1 048.47)?481.6) was higher than that in IGT (760.1?332.1) (P

Objective:To investigate the effects of the β-casomorphin-7 on in vitro and in vivo mice splenic lymphocyte and macrophage nitric oxide production.Methods:Splenocytes proliferation assay and nitrite determination were employed for the studies of effects of β-casomorphin-7 on mice splenic lymphocyte and macrophage following in vitro stimulation of β-casomorphin-7 and in vivo intrapetitoneal administration and drinking β-casomorphin-7 solution administration.Results:In vitro study, β-casomorphin-7 showed a stimulation as well as a suppression of lymphocyte proliferation and significantly(P＜0.01) suppressed the production of nitric oxide. In vivo study, β-casomorphin-7 actions on lymphocytes and macrophages were accordant in intraperitoneal administration and drinking β-casomorphin-7 solution administration. β-casomorphin-7 significantly(P＜0.01) increased in splenic lymphocyte proliferative response and suppressed nitric oxide production in peritoneal macrophages.Conclusion:The present study indicates that the β-casomorphin-7 has the immunomodulatory effects and β-casomorphin-7 may be permeable into peripheral blood intact in mice of 2-3 weeks of age.