BACKGROUND: Basic fibroblast growth factor (bFGF), a kind of polypeptide growth factor possessing multifunctional biological activities,can protect neurons and promote the growth of nerves. It has been corfirmed that bFGF has therapeutic effects on retina ischemia/reperfusion injury (RIRI).OBJECTIVE: To establish RIRI model and analyze the effects of bFGF on cellular apoptosis of retina and the expression of regulatory gene protein.DESIGN: Randomized grouping and validating trial.SETTING: Department of Ophthalmology, the Affiliated Hospital of Medical College of Qingdao University.MATERIALS: The experiment was conducted at the Research Laboratory of Pathology, Department of Ophthalmology, Medical College of Qingdao University, from April 2002 to December 2003. Twenty-eight healthy Wistar rats were enrolled in this experiment. Four rats were randomly chosen for normal control group, the left eyes of the other 24 rats were set as normal saline control group, and the right eyes were set as bFGF group.METHODS: Normal saline control group and bFGF group adopted the rat RIRI models established by transiently elevating intraocular pressure. Normal saline of 12 μL was injected into the vitreous cavity of the left eyes of the rats in normal control group. 12 μL bFGF was injected into the vitreous cavity of the right eyes of the rats in bFGF group, 4 rats once. No administration was given in normal control group. The expression of apoptotic cells was detected and apoptosis indexes were calculated with the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labeling (TUNEL) method and immunohistochemical staining method at the 1st, 6th,12th, 24th,48th and 72nd hours after reperfusion and ischemia for 1 hour.MAIN OUTCOME MEASURES: ① The detection results of apoptotic cells in situ of retina tissuesat different time points after reperfusion. ②The expression of Fas and caspases-2 in retina tissues at different time points after reperfusion.RESULTS ① Comparison of apoptosis indexes of retina tissues at different time points after ischemia reperfusion: There were no apoptotic cells in the retina tissues of the rats in normal control group. As compared with those in normal saline control group, apoptosis indexes in bFGF group were significantly decreased at ischemia 1 hour and reperfusion 1, 6, 12, 24, 48and 72 hours, especially at the 12th, 24th and 48th hours after reperfusion (t =5.362-5.595, P ＜ 0.05). ② The change of Fas expression at different time points after ischemia reperfusion: There was hardly any Fas expression in normal control group. As compared with that in normal saline control group, Fas expression in bFGF group was significantlydecreased at ischemia 1 hour and reperfusion 1, 6, 12, 24, 48 and 72 hours, especially at the 6th, 12th and 24th hours after reperfusion (t=3.954-9.327, P ＜ 0.05). ③The changes of caspase-2 expression at different time points after ischemia reperfusion: There was no caspase-2 expression in normal control group.Compared with that in normal saline control group, the number of caspase2 positive cells in bFGF group was significantly decreased at the 6th,12th,24th, 48th and 72nd hours after ischemia for 1 hour and reperfusion (t=4.125-15.641, P ＜ 0.05).CONCLUSION: bFGF can significantly inhibit the expression of apoptosis gene Fas and caspase-2 in the ischemia and reperfusion of retina, thus reducing cellular apoptosis of ganglion cells and exerting therapeutic effects on the ischemia and reperfusion of retina.

Objective To study the relationship between M3 receptor and prostastic tumor by analyzing the expressions of M3 and vascular endothelial growth factor(VEGF)in adult human normal and neoplastic prostatic gland tissue. Methods The specimens included 36 normal prostates(fresh),36 benign prostatic hyperplasia(BPH)tissue(fresh),and 36 cancer tissue(8 fresh).RT-PCR was used to detect M3 receptor,VEGPs genetic expression.At protein level,VEGF,Ms receptor,CD34 were detected by western-blot and immunohistochemical method. Results VEGF and M3 receptor's genetic expressions were higher in prostate cancer tissue(O.8354±0.1897,0.7824±0.2047)than in BPH tissue(0.6735±0.1603,0.6021±0.1637),while the expressions of these genes were lowest in normal prostate tissue(0.5425±0.1629,0.3436±0.1581)(P<0.001).There was a positive correlation between M3 and VEGFs gene expression(r=0.4999,P

Objective:To prepare poly-L-lactic acid(PLLA)electrospun nanofibers carrying icariin(ICA)(ICA /PLLA)and to evaluate the effects of the ICA /PLLA on MC3T3-E1 cells.Methods:ICA solution was dispersed into PLLA solution,and electrospun fibers were fabricated by W/O emulsion method.The morphology of ICA /PLLA was observed by SEM.The in vitro release kinetics of ICA /PLLA was examined.The attachment of MC3T3-E1 cells on ICA /PLLA was examined by propidiumiodide(PI)labling and ob-served under fluorescent microscope.The proliferation of the cells was measured by MTT assay.The differentiation of the cells was ob-served by alkaline phosphatase (ALP)assay.Results:In vitro,ICA was effectively released from ICA /PLLA for 22 days,cells were attached well on the surface in all groups,ICA did not affect the proliferation of MC3T3-E1 cells(P >0.05),but increased the ALP activity(P <0.05)of the cells.Conclusion:ICA /PLLA can effectively control the release of ICA and promote the differentiation of MC3T3-E1 cells.

Primary hyperparathyroidism (PHPT) in children is a rare condition,co-existing with vitamin D deficiency can lead to more diagnostic uncertainty.Here we report a case of a boy with PHPT complicated with vitamin D deficient rickets.The diagnosis and treatment of this patient was analyzed against literature review to summarize evidence-based clinical features of PHPT in children.We found that compared with adult PHPT,PHPT in children is associated with severer symptoms,higher serum calcium level,lower parathyroid hormone,preponderance of single adenoma in pathology,and higher cure rate of surgery.Co-existence of osteomalacia may induce reduction of the serum calcium level to the normal range,but cause more severe bone lesions.

BACKGROUND:Establishing the animal model of membranous nephropathy is of importance to figure out the pathogenesis of membranous nephropathy.
OBJECTIVE:To investigate the expression of nephrin and podocin in the model of membrane nephropathy in rats, and to investigate their relationships with the pathogenesis of membranous nephropathy.
METHODS:A total of 40 Sprague-Dawley rats were equivalently randomized into model and control groups. Rats in the model group were in premunity by given subcutaneous and multi-point injection of 1 mg cationic bovine serum albumin firstly dissolved in 0.5 mL normal saline and then ful y emulsified with the equal incomplete Freund’s adjuvant for 1 week, and 16 mg/kg cationic bovine serum albumin was injected via vein tails, once every other day for 4 weeks. The same volume of normal saline was injected into the controls. The mRNA expressions of nephrin and podocin in renal tissues were detected using real-time PCR, and biochemical indicators and morphological observation were measured at 2 and 4 weeks after modeling.
RESULTS AND CONCLUSION:(1) In the model group, the total amount of urine and serum albumin levels were significantly decreased accompanying with overt proteinuria, and the serum creatinine, urea nitrogen, cholesterol and triglyceride levels were significantly increased al in a time-independent manner compared with the control group (P<0.01), and the difference was significant (P<0.05). (2) The pathological examination showed that rats in the model group had different degrees of renal tubular dilatation, glomerular basement membrane thickening, mesangial cel s and stromal hyperplasia, which was typical of membranous nephritis. (3) Moreover, the mRNA expressions of podocin and nephrin in the model group were lower than those in the control group. (4) In conclusion, the decreased expressions of podocin and nephfin may disturb the integrity of the slit membrane of podocytes giving rise to the damage of glomerular filtration barrier, and proteinuria appears in final.

Objective:To evaluate the effect of post-conditioning in brain injury induced by myocardial I/R on inflammatory factor and GFAP.Methods:Male Sprague-Dawley rats were randomly allocated into 3 groups (n=8):group Sham,group IR,group IPost.Myocardial IR was induced by occlusion of the anterior descending branch of the left coronary artery for 30 min.group IPost received 3 cycles of 10 s reperfusion followed by 10 s ischemia at the end of myocardial ischemia.The rats were sacrificed at 120 rain of reperfusion and the brains were removed for microscopic examination,inflammatory factors and GFAP.Results:Compared with group Sham,IL-6,IL-8 were significantly increased,IL-10 was down-regulated in group IR(P＜0.01).Post-conditioning can decrease IL-6,IL-8 and up-regulated IL-10(P＜0.01).When compared with group Sham,the expression of GFAP was higher in group IR(P＜0.05),however,the GFAP in group IPost is the most among these three groups(P＜0.01).Conclusion:Post-conditioning could protect brain by decreasing inflammatory factors,increasing GFAP,which both from brain injury induced by myocardial ischemia reperfusion.

Objective To investigate the relationship between serum adiponectin and metabolic syndrome(MS) in children and adolescents with obesity and nonalcoholic fatty liver disease(NAFLD).Methods Four elementary schools and 4 middle schools were selected from Haidian district in Beijing with representative cluster sampling.Two hundred and eighty obese children(obese group),65 obese children with NAFLD(NAFLD group) and 264 normal weight children(healthy control group) aged 7 to 18 years were recruited from the 8 schools with uncompletely randomized sampling.Data including questionnaire,anthropometric measurements,B type ultrasonographic examination for liver were collected and fasting blood laboratory assay were determined.Variables including triglyceride(TG),adiponectin,alanine aminotransferase(ALT) and aspartate aminotransferase(AST) were skewed distribution and natural logarithmical transformations were performed.Chi-square test for category and multiple binary Logistic regression analysis were used to statistical analysis.Results Body mass index(BMI) and waist circumference(WC) in obese group and NAFLD group were higher than those in healthy control group.All the chi-square tests for trend among the 3 groups were statistically significant(P

OBJECTIVETo explore the effect of ERK1/2 MAPK pathway on the expression of Kv1.5 channel, a voltage-gated potassium ion channel, in rat pulmonary artery smooth muscle cells (PASMCs) and its mechanisms during the process of hypoxia.

METHODSThe PASMCs derived from SD rats were cultivated primarily. The third to sixth generation of PASMCs were divided into 5 groups randomly: (1) Normal group (N); (2) Hypoxic group (H); (3) Demethy sulfoxide(DMSO) group (HD); (4) U0126 group (HU): 10 micromol/L U0126; (5) Anisomycin group (HA): 10 micromol/L anisomycin. There were three dishes of cells in each group. The cells in normal group were cultured in normoxic incubator (5% CO2, 37 degrees C), the cells in other groups were added to 0.05% DMSO in the hypoxic incubator (5% CO2, 2% O2, 37 degrees C), all cells were cultured for 60 h. RT-PCR and Western blot were used to detected the espressions of Kv1.5 mRNA and protein in PASMCs.

RESULTSCompared with N group, the expressions of Kv1.5 mRNA and protein in H, HD and HA groups were reduced significantly (P < 0.05); Compared with H group and HD groups, Kv1.5 mRNA and protein expressions in HU group were increased sharply (P < 0.05). Compared with the HU group, Kv1.5 mRNA and protein expressions in HA groups were significantly lower (P < 0.05).

CONCLUSIONLow oxygen reduced Kv1.5 mRNA and protein expressions, U0126 could resistant the Kv1.5 channel lower expression caused by hypoxia. Anisomycin had no significant effect on Kv1.5 channel expression under hypoxia, but the expression of Kv1.5 was still significantly lower than the normal oxygen group. These data suggest that hypoxia may cause hypoxic pulmonary hypertension by interfering ERK1/2 signaling pathway to inhibit Kv1.5

Animals ; Cell Hypoxia ; Hypertension, Pulmonary ; Kv1.5 Potassium Channel ; metabolism ; MAP Kinase Signaling System ; Mitogen-Activated Protein Kinase 1 ; metabolism ; Mitogen-Activated Protein Kinase 3 ; metabolism ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; metabolism ; Oxygen ; Pulmonary Artery ; cytology ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo study the correlation between arachidonic acid (AA) and acute red blood cells damage in rats, and to build a model with hidden blood loss in vivo, and to explore the pathological mechenism of hidden blood loss.

METHODSA total of 50 male adult Sprague-Dawley rats weighing (200 ± 20) g were randomly divided into five groups (n = 10): control group and four experimental groups. The rats in the experimental groups were given 0.5 ml different concentrations of AA dilu- ents, 5, 10, 20, 40 mmol/L respectively. The blood samples were collected from orbital venous at the beginning and 24, 48, 72 hours after administration. Then the changes of hemoglobin (Hb) ,red blood cell count (RBC), glutathione peroxidase (GSH- PX) activity, total superoxide dismutase (T-SOD) activity and hydrogen peroxide (H202) in the blood samples were tested.

RESULTSSignificant hidden blood loss occurred when the concentration was 10 mmol/L in the experimental group, with the RBC and Hb sharply reduced in blood samples. The Hb and RBC were reduced in all the experimental groups and control group at 24 hours after administration, while in the experimental groups they changed more obviously. The GSH-PX activity, T-SOD activity and H₂O₂were also significantly reduced in all groups, and the changes showed significant differences. The Hb and RBC were relatively stable in the control group and the experimental groups at 48 hours after administration; while GSH-PX activity, T-SOD activity and H₂O₂were all significantly decreased, and the changes in the experimental groups were more notable.

CONCLUSIONElevated levels of AA in the blood causes oxidative stress in the red blood cells, leading to the damage of red blood cells and hemoglobin, which is responsible for hidden blood loss.

Animals ; Arachidonic Acid ; toxicity ; Erythrocytes ; drug effects ; metabolism ; Glutathione Peroxidase ; blood ; Hemoglobins ; analysis ; Male ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; blood

Objective Studying the status quo and constraints for rural healthcare service in grassroot rural healthcare units,for policy recommendations.Methods Using data from the fourth healthcare service investigation,by means of quantitative interview and qualitative interview,for an investigative interview of 348 township hospitals and 251 village clinics in 31 provinces in China.Results Deployment percentage of primary heahhcare services at township hospitals level is 49.1% (28.0/57),and that for village clinics is 60.6%(5.4/9).Conclusions The key to upgrading rural healthcare service system at grassroots level is to deploy better diagnostic equipments,upgrade the diagnostic competence of grassroots healthcare personnel and build a continuous service system for primary healthcare service.