ObjectiveTo observe the effects of basic fibroblast growth factor (bFGF) on growth period of cultured bovine retinal pigment epithelium cells in different durations.MethodsDifferent proportion of phase with 4 ng/ml bFGF were measured by Flow Cytometry (FCM) assay.ResultsbFGF increased the RPE cells' proportion of phase S and decreased the proportion of phase G0-G1. ConclusionbFGF can stimulate RPE cells from phase G1 to phase S.

Administration, Inhalation ; Bronchiolitis ; drug therapy ; Bronchodilator Agents ; administration & dosage ; therapeutic use ; Budesonide ; administration & dosage ; therapeutic use ; Female ; Humans ; Infant ; Male ; Poly I-C ; administration & dosage ; therapeutic use

AIM: To optimize the conditions for in vitro culture of retinal pigment epithelium (RPE) cells, we characterized expressions of various growth factors in RPE cells, including tumor necrosis factor (TNF-α), vascular endothelial growth factor (VEGF), β fibroblast growth factor (βFGF), transforming growth factor β2 (TGFβ2), and interferon-γ (IFN-γ). We also studied expressions of caspase-3 under different concentrations of fetal bovine serum (FBS) with insulin-transferrin-sodium selenite (ITS) supplement. METHODS: First, we investigated if expressions of TNF-α, VEGF, βFGF, TGFβ2, IFN-γ, and caspase-3 in FBS and ITS with of concentration. Second, we cultured primary RPE cells from eyes of forty C57 BL/6 mice in standard dulbecco's modified eagle's medium (DMEM) containing 20,40,100mL/L FBS and 20,40,100mL/L FBS together with 10g/L ITS. Immunohisto-chemical staining and cell counting were performed to verify the existence and growth condition of RPE cells. Expressions of TNF-α, VEGF, βFGF, TGFβ2 and IFN-γ were determined using cells and supernatant from passage-3 to -4 primary RPE cell after 48 hours of culture with RT-PCR and enzyme-linked immunosorbent assays (ELISA). The expression of casepase-3 was determined via Western blotting. The major outcome measurement is the expression level of growth factors in cultured RPE cells and the experiment design is to expose the RPE cells to different culture medium. RESULTS: TNF-α, VEGF, βFGF, TGFβ2, but not IFN-γ, were expressed and the expressions increased with concentration. No expression of the aforementioned genes was detected in presence of ITS. The primary cultures of RPE cells were successfully established. TNF-α, VEGF, βFGF, TGFβ2 (but no IFN-γ) and the active caspase-3 were detected in 20,40,100mL/L FBS or 20,40,100mL/L FBS combined with 10g/L ITS; the expressions were upregulated with increasing concentration of FBS. There is no significant difference in the expression of growth factors between these groups. However, significant differences were shown among different concentration of FBS (P＜0.01）. The lowest expression was observed in 20mL/L FBS or 20mL/L FBS combined with 10g/L ITS medium with RPE cells. But RPE cells were shown in better growth condition in 20mL/L FBS combined with 10g/L ITS.CONCLUSION: TNF-α, VEGF, βFGF, TGFβ2 and caspase-3 were expressed in RPE cells and supernatants. The production of above 20mL/L FBS combined with 10g/L ITS in DMEM may be the ideal cell culture medium that supports the normal growth of RPE cells.

OBJECTIVETo understand the status of occupational health of female migrant workers in different kinds of pharmaceutical industries in Gansu province and to provide the basis for improving occupational health condition.

METHODSOne thousand eight hundreds and one female workers from 16 enterprises were selected by cluster sampling in Gansu province and investigated by interviewing and questionnaires.

RESULTSThere were statistical significances of education level, status of residency registrations, employment relationship and occupational hazards among female workers in three types of enterprises (P < 0.05 or P < 0.01). The morbidities of skin disease in female workers for three kinds of enterprises were 4.46%, 2.53% and 3.70%, respectively. The morbidities of reproductive system disease in female workers for three kinds of enterprises were 48.57%, 36.70% and 36.11%, respectively.

CONCLUSIONThe levels of education and working conditions of female workers in the traditional Chinese medicine, western medicine plants are low. There are more severe occupational hazards in female workers of the traditional Chinese medicine plants.

Adolescent ; Adult ; Drug Industry ; Female ; Health Status ; Humans ; Middle Aged ; Occupational Exposure ; Occupational Health ; Surveys and Questionnaires ; Transients and Migrants ; Young Adult

The aim of the study is to establish a new method of quality evaluation and validate its feasibilities by the simultaneous quantitative assay of four lignanoids in Schisandra chinensis. A new quality evaluation method, quantitative analysis of multi-components by single marker (QAMS), was established and validated with Schisandra chinensis. Four main lignanoids, schisandrin, schisantherin A, deoxyschizandrin and gamma-schizandrin, were selected as analytes and schisandrin as internal reference substance to evaluate the quality. Their contents in 13 different batches of samples, collected from different bathes, were determined by both external standard method and QAMS. The method was evaluated by comparison of the quantitative results between external standard method and QAMS. No significant differences were found in the quantitative results of four lignanoids in 13 batches of S. chinensis determined by external standard method and QAMS. QAMS is feasible for determination of four lignanoids simultaneously when some authentic standard substances were unavailable, and the developed method can be used for quality control of S. chinensis.
Chromatography, High Pressure Liquid ; methods ; Cyclooctanes ; analysis ; Dioxoles ; analysis ; Drugs, Chinese Herbal ; chemistry ; Fruit ; chemistry ; Lignans ; analysis ; Plants, Medicinal ; chemistry ; Polycyclic Compounds ; analysis ; Quality Control ; Schisandra ; chemistry

BACKGROUNDCataracts is considered be formed because of an abnormal glucose metabolic pathway or oxidative stress. We explored the damaging role of ONOO- and antagonism of cholecystokinin octapeptide-8 (CCK-8) in diabetic cataractal rat lenses.

METHODSA diabetic cataractal animal model was established by peritoneal injection of streptozotocine (STZ). Thirty-six normal SD rats were taken as control group; seventy-two were given STZ (45 mg/kg) and then divided into STZ group and CCK-8 group (peritoneal injection CCK-8). STZ induced diabetic rats were treated with CCK-8 for 60 days. Lenses were examined with slit lamp at 20, 40 and 60 days. Immunofluorescent staining and Western blot analysis were used for determining nitrotyrosine (NT, a marker for ONOO-). PT-PCR and gene array analysis were used for determining the expression of inducible nitric oxide synthetase mRNA (iNOS mRNA) in lens epithelium (LEC).

RESULTSSTZ group rats developed lens opacity by 20 days that reached a high level by 60 days after STZ injection. CCK-8 group rats delayed the cataract formation. CCK-8 group rats delayed the cataract formation. There was no distinct expression of NT and iNOS mRNA in control group. In STZ group, there were distinct expression of NT and upregulation of iNOS mRNA; however, CCK-8 group showed weak expression of NT and downregulation of iNOS mRNA.

CONCLUSIONSNT, which may be a new form of oxidative stress, was expressed in diabetic rat LEC although CCK-8 could reverse NT damage in LEC. The results suggested that CCK-8 might be a useful therapeutic agent against diabetic cataract. The antagonizing mechanism of CCK-8 may be related to direct antagonism of ONOO- as well as its inhibition of the expression of iNOS mRNA for production of NO and therefore decrease in the formation of ONOO-.

Animals ; Blotting, Western ; Cataract ; etiology ; prevention & control ; Diabetes Mellitus, Experimental ; complications ; Fluorescent Antibody Technique ; Male ; Nitric Oxide Synthase Type II ; genetics ; Oxidation-Reduction ; Peroxynitrous Acid ; metabolism ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction ; Sincalide ; pharmacology ; Streptozocin ; Tyrosine ; analogs & derivatives ; genetics

OBJECTIVETo determine whether blood lipids profile, fibrinogen and viscosity were associated with passive smoking (i. e. environmental tobacco smoke, ETS) in Chinese women who never smoke.

METHODSIn Xi'an, China, a case-control study was carried out on 115 cases of coronary heart disease (CHD) defined by coronary arteriography (CAG) and 208 non-CHD controls confirmed by CAG and/or exercise electrocardiography. Data on exposure to ETS, defined as exposure from cigarettes smoking husband or co-workers or both for at least 5 years, was obtained through standardized interviews. Standard laboratory methods were used and the lipid measurements were under US CDC quality control programs.

RESULTSIn the subjects defined by CAG, the levels of high density lipoprotein cholesterol (HDL-C), HDL2C, apolipoprotein (apo) A1 among passive smokers appeared lower than those in non-passive smokers,but the low density lipoprotein cholesterol (LDL-C), apoB, apoB/A1, fibrinogen, plasma and whole blood viscosity were higher than that in non-passive smokers. There were positive associations of the numbers of coronary arteriosclerosis with the levels of blood lipids,fibrinogen and viscosity. In the non-CHD controls, 81 subjects were not exposed and 127 were exposed to ETS. The P values of t-test for the adjusted (for age, body mass index, present diseases history) means between two groups were listed below: 0.06 (total cholesterol), 0.30 (triglyceride), 0.004 (HDL-C), <0.001 (HDL2-C), < 0.001 (apoA1), 0.009 (apoB), <0.001 (apoB/apoA1), <0.001 (fibrinogen), <0.001 (plasma viscosity), <0.001 and 0.004 [two measures (5.75/s and 230/s) of whole blood viscosity]. The correlation coefficients between cumulative exposure of passive smoking and HDL-C,HDL2-C,apoA1, apoB, apoB/apoA1, fibrinogen, plasma viscosity, and two measures of whole blood viscosity were -0.25, -0.27, -0.30, 0.24, 0.31, 0.32, 0.43, 0.51 and 0.36 (all P<0.01), respectively.

CONCLUSIONPassive smoking could affect blood lipid metabolism, fibrinogen and viscosity in the never smoking women which might contribute to the causation of coronary heart disease.

Apolipoprotein A-I ; blood ; Blood Viscosity ; drug effects ; Cholesterol, HDL ; blood ; Cholesterol, LDL ; blood ; Coronary Disease ; etiology ; Female ; Fibrinogen ; metabolism ; Humans ; Male ; Middle Aged ; Tobacco Smoke Pollution ; adverse effects

OBJECTIVETo study the changes of vascular endothelial cell function and platelet activation in rabbit spinal cord following ischemia-reperfusion (I/R) injury and their roles in the spinal cord injury.

METHODSRabbit spinal cord I/R injury models were established using Zivin method, and the changes in plasma NO and GMP140 levels were dynamically monitored after the injury.

RESULTSPlasma NO level increased significantly in the I/R group at the end of the ischemia, and reached the peak level at 2 h of reperfusion as compared to that in sham-operated group (P<0.01). Plasma NO level decreased at 6 h of reperfusion, but still significantly higher than the level in the sham-operated group (P<0.05). Plasma GMP140 underwent no significant changes in the sham-operated group, but significantly increased in the I/R group at the end of the ischemia, followed by gradual declination to the normal level at 2 h of reperfusion.

CONCLUSIONSpinal cord I/R injury causes overexpressions of NO and GMP140, suggesting the involvement of endothelial cell injury and platelet overactivation in the pathological process and repair of spinal cord I/R injury.

Animals ; Endothelial Cells ; metabolism ; Nitric Oxide ; blood ; P-Selectin ; blood ; Platelet Activation ; Rabbits ; Reperfusion Injury ; blood ; pathology ; physiopathology ; Spinal Cord ; pathology ; physiopathology

The present study was designed to observe if puerarin decreases lens epithelium cell (LEC) apoptosis induced partly by peroxynitrite (ONOO(-)). One hundred and eight rats were randomly divided into control group (n=36), streptozotocin (STZ) group (n=36) and STZ + puerarin group (n=36). The rats in the control group intraperitoneally (i.p.) received 0.5 ml of saline. The rats in STZ group and STZ + puerarin group received intraperitoneal injection of STZ (45 mg/kg). Three days later, the rats in STZ + puerarin group were given puerarin (140 mg/kg per day, i.p.). On days 20, 40 and 60 of the experiment, morphologic changes of lenses were observed with slit lamp. Then the animals were sacrificed for further analysis. The amount and percentage of apoptotic LECs were determined by flow cytometry. Nitrotyrosine (NT, the foot print of ONOO(-)) was examined by immunohistochemistry. Apoptosis-related genes (iNOS, etc.) were analyzed by gene array. The results showed that in the control group, all the lenses were clear. In STZ group, gradually severe opacity of the lens was observed on days 20, 40 and 60. But in STZ + puerarin group, mild opacity of the lens was observed on day 20 and more severe on day 40, but markedly decreased on day 60. In the control group, mild apoptosis of LECs was observed. In STZ group, time-dependent increase in apoptosis of LECs was observed. In STZ + puerarin group, mild apoptosis of LECs was observed on day 20, significantly increased on day 40, but markedly decreased on day 60. There was no expression of NT in the lens in the control group, but an increased expression of NT in STZ group. In STZ + puerarin group, mild expression of NT was observed on day 20, significantly increased on day 40, but markedly decreased on day 60. There was no expression of iNOS in the lens in the control group, but continuous up-regulation of iNOS expression in STZ group. In STZ + puerarin group, mild expression of iNOS was observed on day 20, significantly increased on day 40, but markedly decreased on day 60. Except the changes of iNOS related to NO production, the other apoptosis-related genes, including BCL-2 and SOD were down-regulated, while NF-kappaB and TNFR1-FADD-caspase signal transduction way were up-regulated in STZ group. The results were opposite in STZ + puerarin group and the control group. These findings show that NT is expressed in diabetic rat lens, which proves that LEC apoptosis in diabetic lens is partly induced by ONOO(-) which may be a new oxidative damage way to form cataract. Puerarin partly decreases LEC apoptosis induced by ONOO(-) and is a potential medicine for therapy of diabetic cataract. The mechanism of puerarin dealing with diabetic cataract may be related to its direct inhibition of LEC apoptosis and antagonism of ONOO(-) in diabetic rats.
Animals ; Apoptosis ; Cataract ; chemically induced ; Diabetes Mellitus, Experimental ; Epithelial Cells ; drug effects ; Isoflavones ; pharmacology ; Lens, Crystalline ; cytology ; Nitric Oxide Synthase Type II ; metabolism ; Peroxynitrous Acid ; Rats ; Tyrosine ; analogs & derivatives ; metabolism

OBJECTIVETo prospectively study the relationship between risk factors and stroke incidence and mortality in the Chinese elderly.

METHODSAn analytic study in a cohort population of 1,268 male retired cadres in a military setting. A health-screening program was carried out for all cadres aged 55 years or older in Xi'an in February 1987. Baseline data on smoking, cholesterol, triglyceride, blood pressure, body mass index (BMI), histories of hypertension, hyperlipidemia and coronary heart disease (CHD), stroke, diabetes and family histories of cardiovascular disease were investigated. Main outcome measures were stroke incidence, stroke and all-cause mortality.

RESULTSThe cohort was followed up until June 30, 2001 and a total number of follow-up person-year was 15,546. During the follow-up period, there were 113 new stroke cases identified and the adjusted incidence was 727 per 100,000 person-year. Forty-five deaths were due to stroke and the adjusted mortality was 289 per 100,000 person-year. Using Cox model analysis, after adjustment on age, total cholesterol, triglyceride, smoking, drinking and physical exercise, we noticed that the systolic pressure, BMI, history of CHD and hyperlipidemia were independent risk factors for stroke incidence and morality.

CONCLUSIONThe incidence and mortality of stroke in this cohort were lower than those in the same age group of general population. Monitoring and controlling body mass index and blood pressure level seemed to be important factors for the prevention of stroke in the elderly.

Aged ; Aged, 80 and over ; Blood Pressure ; Body Mass Index ; Cohort Studies ; Coronary Disease ; complications ; Humans ; Hyperlipidemias ; complications ; Hypertension ; complications ; Incidence ; Male ; Middle Aged ; Military Personnel ; Risk Factors ; Stroke ; epidemiology ; etiology ; mortality