BACKGROUND:α-zearalanol, a natural phytoestrogen has the effect of anti- atherosclerosis like the estrogen but with less side effect. Therefore, it has a potential for more application in the future. OBJECTIVE: To investigate the effect of 17β-estradiol (E2) and α-zearalanol (ZAL) on coagulation and fibrinolysis in ovariectomized rats and to compare and analyze their effects. DESIGN: An observational and controlled experiment. SETTING:Department of Pathophysiology of the Capital University of Medical Sciences and the Department of Pathophsiology of the Institute of Basic Medical Sciences of Chinese Academy of Medical Sciences and Peking Union Medical College. MATERIALS: The experiment was completed at the Science Department of Experiment Animals from July to September 2003. Thirty-six healthy female Wistar rats aged 12-week old , weighing (250±10)g, clean grade were involved. The animals were divided into 4 groups, namely sham-operation control group, ovariectomy (OVX) group, OVX +E2 group, OVX+ZAL group with 9 rats in each group. METHODS: For the rats in the sham-operation control group, operation was performed without removing the ovary. For the rats in the OVX group,ovariectomy was performed and the rat models were made at the sterile condition. 1 mg/kg 17β-estradiol and 1mg/kg α-zearalanol were respectively injected intramuscularly into the rats in the OVX +E2 group and OVX+ZAL group 14 days after the operation, once every three days for 35days altogether. After the administration of 17β-estradiol or α-zearalanol for 5 weeks, rats were killed, and blood was collected through the common carotid artery and plasma was collected from it. The prothrombin time (PT)and the activated partial thromboplastin time (APTT) were determined with coagulation method. The level of fibrinogen (FG) was measured with an automatic biochemistry analyzer. Tissue factor(TF)level was determined with ELISA method, the activity of tissue plasminogen activator(t-PA)and plasminogen activator inhibitor type 1 (PAI-1)were determined with chromogenic substrate assay. At the same time, the uteri were cut off and weighed by an electronic scale to work out the uteri weight/body weight (g/kg). MAIN OUTCOME MEASURES: ① PT, APTT, FG,TF, t-PA, PAI-1; ②uteri mass/body mass RESULTS: All the 36 rats entered the stage of the result analysis. ①change of PT: it was shorter in the OVX group than that in the sham-operation control group (P ＜ 0.01 ), but it was longer than that in the OVX group after supplementation of E2 and ZAL (P ＜ 0.05-0.01 ). ② Change of FG and TF: they were significantly higher than those in the sham-operation control group (P ＜ 0.05-0.01 ). But, they were lower than those in the OVX group after supplementation of E2 and ZAL (P＜ 0.05-0.01 ). ③Change of tPA: It was significantly lower in the OVX group than that in the sham-operation control group [(0.33±0.33) μkat/L,(4.00±1.50) μkat/L,(q=9.43, P ＜ 0.01 )]. However, it was significantly higher than that in the OVX group after supplementation of E2 and ZAL [(1.83 ±0.67)μkat/L,(1.17±0.83) μkat/L, (q=13.50, P ＜ 0.01; q=5.00, P ＜ 0.05). ④ Change of PAI-1: It was significantly higher than that in the sham-operation control group [(2.33±0.67) μkat/L,(1.17±0.33) μkat/L,(q=10.5, P ＜ 0.01 )]. ⑤Uteri mass/body mass: It was significantly lower in the OYX +ZAL group than that in the OVX+ E2 group [0.66,1.96, (q=14.67, P ＜ 0.01)]. CONCLUSION: Both 17β-estradiol and α-zearalanol can resume the balance of coagulation-fibrinolysis of ovariectomized rats, suggesting that αzearalanol has a similar protective effect similar to that of 17β-estradiol on cardiovascular system. As α-zearalanol has less adverse effect on uteri enlarging than 17β-estradiol, it has a better prospect for substitution of estrogens as a natural phytoestrogen.

AIM: To study the influence of minimal modified low density lipoprotein (mmLDL) on plasminogen activator inhibitor-1 (PAI-1) activity, gene expression and regulation in human vascular endothelial cells. METHODS: The PAI-1 activity in human umbilical vein endothelial cells (HUVEC) culture medium was measured by chromogenic assay. The PAI-1 mRNA expression were determined by Northern blot. Using gene recombination techniques, four luciferase reporter gene plasmids containing different length of human PAI-1 gene promoter were constructed. Through the transient transfection analysis, the roles of AP-1 element(from -823 bp to -553 bp) in PAI-1 promoter have been determined. In order to further verify the role of AP-1 element, the three site-directed mutants were received using PCR and sequencing assay. RESULTS: The PAI-1 activity and mRNA level were increased when HUVECs were exposed to 50 mg/L mmLDL. At the same time, the AP-1 protein level was increased in nuclear. The induction by mmLDL were decreased markedly when the three AP-1 elements in PAI-1 promoter had been mutated, respectively. CONCLUSION: (1) mmLDL increased PAI-1 activity and mRNA expression in HUVEC. (2) Increase in PAI-1 activity induced by mmLDL was related to its mRNA expression. (3) Three AP-1 element in PAI-1 promoter may have an important role in PAI-1 gene transcription in endothelial cells induced by mmLDL.

An economic and facilely prepared dopamine ( DA) sensor have been successfully fabricated by the electrodeposition of copper on single-walled carbon nanotubes ( SWNTs )/Nafion-modified glassy carbon electrode. The morphology of the material was observed by scanning electron microscopy ( SEM) and element composition of the material was investigated by energy dispersive X-ray spectroscopy ( EDX ) . Tests with various scan rates and pH conditions indicated that an adsorption-controlled process occured in the electrochemical system. The mechanism of the electrode reaction of dopamine involved a two-electron process which was accompanied by a deprotonation step. Electrochemical parameters were calculated with the electron transfer number as 2 . 67 , the charge transfer coefficients as 0 . 6 , the apparent heterogeneous electron transfer rate constant as 1. 38 s-1 . Under the optimal conditions with differential pulse voltammetric measurement, the linear equation was Ipa(μA)=-0. 054c (μmol/L)-3. 82(R2=0. 9988), with linear range of 5-100 μmol/L and detection limit of 0 . 0135 μmol/L ( S/N=3 ) . The main advantages of sensor included facile fabrication approach, high sensitivity, good stability and high reproducibility. The sensor was applied to the detection of DA in volunteer urine by differential pulse voltammetry with favorable recoveries of 96 . 5%-100 . 4% and relative standard deviations (RSDs) of 1. 2%-2. 4%.

Objective To observe the effect of superoxide dismutase ( SOD ) , malondialdehyde ( MDA ) , glutathione (GSH), catalase (CAT) and endothelin (ET-1) and tumor necrosis factor alpha (TNF-α) on rat renal tissue under acute hypoxia .Methods 24 male Wistar rats, weight 180~220 g, were randomly divided into control group and acute hypobaric hypoxia group .Acute hypoxia group was divided into 2 groups hypoxia 1 and hypoxia 2, 8 rats for each group.After acute hypobaric hypoxia 10min and 24h, rats were sacrificed.The left removed kidneys were analyzed for biochemical indexes , and the right parts were observed by immunohistochemistry to evaluate the expression level of renal endothelin (ET-1) and tumor necrosis factor alpha (TNF-α).Results After acute hypobaric hypoxia , the activity of SOD of the rats kidney was greatly decreased (P <0.01), CAT activity of hypoxia group 1 was significantly decreased (P <0.01), GSH activity of hypoxia group 2 was significantly decreased (P <0.05), but the MDA content had no obvious change ( P >0.05).The immunohistochemical staining showed that , the expression level of ET-1 and TNF-αwas increased remarkably, but it was reduced after 24 h.Conclusion The obviously decreased activity of SOD , CAT, GSH and significantly increased expression of ET-1 and TNF-α, may be involved in the pathogenesis of renal hypoxic injury .

AIM: To explore the effect of complement on the cerebral ischemia/reperfusion injury in rat and the protection by sCR1-SCR15-18. METHODS: 75 male SD rats were randomly divided into three groups: sham operation group (SO, n=15), middle cerebral artery occlusion and reperfusion (MCAO) without treatment group (I/R, n=30); MCAO treated with sCR1-SCR15-18 group (sCR1-SCR15-18, n=30). After the MCAO for 2 h, then reperfusion for 24 h, the scores of neural behavioral functional deficits were determined. Infarction area was measured by TTC staining. Activity of MPO in cerebral cortex was detected. C3b deposition and pathological change were observed by immunohistochemial staining and HE staining, respectively. RESULTS: After reperfusion for 24 h, the neurological deficits score, infarction area and activity of MPO in sCR1-SCR15-18 group were decreased compared to I/R group. In sCR1-SCR15-18 group, C3b deposition in ischemic area was decreased and pathological injury was improved compared to I/R group. CONCLUSION: Complement plays a role in cerebral ischemia-reperfusion injury and sCR1-SCR15-18 exerts a protective effect by inhibiting the excessive activation of complement.

BACKGROUND: The patients with severe spleen rupture can save their spleen functions by auto-transplantation of the spleen tissues in the greater omentum. Whether the transplanted spleen tissues are regulated by nerves or not is still unclear. The growth associated protein, GAP-43, is a specific protein of the nervous system, and by testing the GAP-43 in the transplanted spleen tissues, nerve regeneration can be understood.OBJECTIVE: To study the mRNA expression of the GAP-43 in the regenerated spleen tissues at different phases after the auto-transplantation of the spleen tissues, and to find the regeneration regularity of the GAP-43 + nerves in the auto-transplanted spleen tissues.DESIGN: A randomized and controlled experiment SETTING: A teaching and research office of the surgical applied anatomy and operative surgery department in a university MATERIALS: The study was done in the Surgical Applied Anatomy and Operative Surgery Department of the Third Military Medical University of Chinese PLA from September 2002 to July 2003. A total of 120 Wistar mice were chosen. The mice were randomly divided into the experiment group and pseudo-operation group(control group) to make animal models. After the operation, mice of the two groups were fed under the same circumstances. The spleen tissues were respectively taken at the 15th, 30th, 60th,90th, 120th and 180th days after the operation for the study.METHODS: The general RNA was extracted from the tissues using the Tripure agent by the routine method. The general RNA was reverse transcribed into cDNA using the M-MLV reverse transcription kit. The mRNA of the GAP-43 was quantitatively measured using the gel pattern analysis.tern analysis.RESULTS: Totally 30 days after the transplantation of the spleen tissues, the mRNA of the GAP-43 was found to be expressed in the auto-transplanted spleen tissues. Ninety days after the operation, the expression reached the peak level. Totally 120 to 180 days after the operation the amount of the mRNA of the GAP-43 in the transplanted spleen tissues gradually got close to that in the normal spleen tissues.CONCLUSION: The expression of mRNA of the GAP-43 in the auto-transplanted spleen tissues suggests the regeneration of the nervous fiber in the transplanted spleen tissues.

Objective To detect the mutations in ATP2C1 gene of 5 sporadic patients with Hailey-Hailey disease (HHD).Methods Five sporadic patients with HHD collected from the outpatient clinic setting were recruited into this study with informed consent.Blood samples were taken from all patients and 100 unrelated human controls.and DNA was extracted from these samples.Mutation scanning was carried out for ATP2C1 gene by polymerase chain reaction (PCR) and direct sequencing.Results The diagnosis of all cases was confirmed by typical clinical manifestation,cutaneous pathology and immunofluorescence pathology.Five novel mutations.including a deletion mutation (2025delG),three missence mutations (L269R,C348R,A651D) and a non-sense mutation (Q259X) were identified in these cases.No mutations were detected in any of the 100 controls.Conclusion Five novel mutations in ATP2C1 gene have been identified for Hailey-Hailey disease.

Objective Toexplore the correlation of ApoE gene polymorphism and serum uric acid levels in Ningxia Hui Autonomous Region , China.Methods A case-control study.October 2011 to November 2011, five hundred twenty eight ( 296 male, 232 female ) apparently healthy individuals were studied.Questionnaires and physical examinations were performed by standard operation procedure.Fasting blood was collected for biochemistry testing including serum lipid parameters , uric acid concentration and creatinine levels.The multi-ARMS PCR was applied to determine ApoE genotypes ,and the relation of ApoE genotypes with serum lipid parameters and uric acid levels were analyzed.Non-normal distribution were compared using cause and inspection.Results The common six kinds of ApoE genotype can be detected.The total cholesterol ( TC) ,low density lipoprotein cholesterol ( LDL-C) and uric acid ( UA) levels in different genotype subgroups had statistical differences.The individuals with ε2/3 genotype had a significantly greater reductions in TC and LDL-C levels but increment in uric acid concentration than those withε3/3 and ε3/4 genotype (P<0.05).The effect of ApoE gene polymorphism on uric acid levels still remained significantly after adjustment for age , gender , region and other factors.Conclusion The ApoE polymorphism is associated with serum uric acid levels and individuals with ε2 allele have higher serum uric acid levels.

The aim of this study was to investigate the expression of circulating microRNAs (miRNAs) in apolipoprotein E (apoE) knockout mice (apoE-/-) and to validate the role of these miRNAs in human coronary artery disease (CAD). Pooled plasma from 10 apoE-/- mice and 10 healthy C57BL/6 (B6) mice was used to perform the microarray analysis. The results showed that miR-34a, miR-21, miR-23a, miR-30a and miR-106b were differentially expressed in apoE-/- mice, and these expression changes were confirmed by real-time quantitative reverse-transcription PCR. Then, miR-34a, miR-21, miR-23a, miR-30a and miR-106b were detected in the plasma of 32 patients with CAD and of 20 healthy controls. Only miR-34a, miR-21 and miR-23a were significantly differentially expressed in the plasma of CAD patients (all P<0.01). In conclusion, miR-34a, miR-21 and miR-23a were elevated in CAD patients, which means that these miRNAs might serve as biomarkers of CAD development and progression.
Aged ; Animals ; Apolipoproteins E/deficiency ; Biomarkers ; Case-Control Studies ; Coronary Artery Disease/*genetics ; Disease Models, Animal ; Gene Expression Profiling ; Gene Expression Regulation ; Humans ; Male ; Mice ; Mice, Knockout ; MicroRNAs/*genetics ; Middle Aged ; Pilot Projects ; Reproducibility of Results ; Risk Factors

OBJECTIVETo assess the feasibility of using saliva for Schistosomiasis japonica diagnosis.

METHODSSchistosoma japonicum infected animal model was established. Pairs of saliva and serum samples from rabbits and chronic schistosomiasis patients were collected. Anti-schistosoma specific antibodies in saliva and serum were detected by indirect ELISA.

RESULTSThe specificities of antibody detection of rabbit saliva and serum were 93% (28/30) and 97% (29/30), respectively, and the sensitivities of antibody detection of rabbit serum and saliva were 100% (24/24) and 88% (21/24), respectively. A significant correlation (r = 0.5307, P = 0.0038 < 0.05) existed between anti-SEA IgG levels in serum and saliva. As with those in serum, anti-SEA IgG levels in saliva could reflect the state of infection and treatment. The sensitivity of antibody detection was 91% (29/32) for patient saliva samples and 100% (32/32) for their sera. 8 samples were positive in 140 normal saliva samples (i.e. 6% false positive rate) and 6 samples were positive in 156 normal serum samples (4% false positive rate). There was a significant correlation (r = 0.4227, P = 0.008 < 0.05) between specific antibodies in saliva and serum.

CONCLUSIONThe detection of specific antibodies in saliva can be used as a non-invasive immunodiagnosis method of Schistosomiasis japonica.

Adolescent ; Adult ; Animals ; Antibodies, Helminth ; analysis ; Child ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Immunoglobulin G ; analysis ; Male ; Middle Aged ; Rabbits ; Saliva ; immunology ; Schistosoma japonicum ; immunology ; Schistosomiasis japonica ; diagnosis