Objective To explore the effect of oxidative stress on human Wiskott-Aldrich syndrome related protein 2 (WAVE2) expression in placental trophoblasts in women with preeclampsia.Methods (1) Twenty women with preeclampsia and twenty-three normal term pregnant women,delivered from August 15,2011 to February 23,2012 in the First Affiliated Hospital of Chongqing Medical University,were recruited and divided into preeclampsia group and control group.Placenta samples were collected after cesarean section.The localization and distribution of WAVE2 in placenta was studied by immunohistochemistry.Quantitative real-time polymerase chain reaction and Western blot were employed to assay the WAVE2 mRNA and protein levels.Tissue homogenates was applied to determine the levels of reactive oxygen species (ROS).The correlation between ROS levels and WAVE2 was also analyzed.(2) An in vitro hypoxia/reoxygenation (H/R) model was utilized to simulate ischemia/reperfusion injury to placental trophoblasts.The HTR-8/ SVneo cells (immortalized human first trimester extravillous trophoblast cells) were pre-incubated overnight,after exposure to H/R or normoxic conditions for 48 hours.Flow cytometry was employed to analyze intracellular ROS level.Meanwhile,Transwell assay was utilized to analyze the invasion and migration of HTR-8/SVneo cells.The location and expression of WAVE2 in trophoblasts was evaluated by cell immunofluorescence and Western blot.Statistical differences between the two groups were evaluated by independent t-tests.Pearson's correlation coefficient test was used for correlation analysis.Results (1)Compared with the control group,preeclampsia group had significantly higher 24-hour proteinuria [(1.96±0.24) g vs (0.08±0.05) g,t=19.436,P＜0.05],systolic blood pressure [(154 ± 13) mm Hg vs (98 ±11) mm Hg,t=11.324,P＜0.05] and diastolic blood pressure [(105±14) mm Hgvs (69±8) mm Hg,t=9.612,P＜0.05].In addition,the placental weight and birth weight of infants in preeclampsia group were significantly reduced as compared to the control group [(432±53) g vs (536±67) g,(2446± 187) g vs (3207± 233) g,t=14.562 and 16.307,allP＜0.05)].The WAVE2 mRNA level (0.28±0.07 vs 1.01±0.02,t=12.747,P＜0.05) and the WAVE2 protein levels (0.63±0.08 vs 1.34±0.19,t=11.648,P＜0.05) were also significantly decreased in preeclampsia groups.The level of ROS in placenta in the preeclampsia group was significantly higher than in control group [(144.22 ± 12.32) nmol/(mg · prot) vs (75.17 ± 8.71) nmol/(mg · prot),t=20.467,P＜0.05].There was significant negative correlation between ROS level and WAVE2 protein expression in preeclamptic placenta (r =-0.726,P =0.000).(2) In vitro study showed that,the levels of ROS in normoxia group and H/R group was (82.9±5.8)％ and (155.6±8.1)％,(t=12.747,P＜0.05).Compared with normoxia condition,decreased cell invasion and migration were found in HTR-8/SVneo cells in H/R group [(51.9 ± 3.3)％ and (58.4 ±4.2)％ respectively,t=11.034 and 13.839,P＜0.05].Results from the cell immunofluorescence showed that WAVE2 protein located in the cytoplasm of HTR-8/SVneo cells,and the expression of WAVE2 protein was significant decreased in HTR-8/SVneo cells after exposure to H/R for 48 h (0.37±0.05 vs 0.76±0.06,t=8.631,P＜0.05).Conclusions Excessive oxidative stress in preeclamptic placentas was correlated with the decreased expression of WAVE2.H/R-induced oxidative stress could decrease WAVE2 expression,which may contribute to impaired trophoblast invasion and migration in preeclampsia.

Objective:To observe HSP70 expression during experimental tooth movement in rats.Methods:Orthodontic appliance was placed between the maxillary right first molar and maxillary central incisors of 30 adult SD rats.The rats were killed at 1,3,5,7,14 d after orthodontic force application,samples of experimental teeth with periodontal tissue were prepared for immunohistochemical examination of HSP70 expression.Result:Strong expression of HSP 70 in periodontal ligment was observed at 1 and 3 d after orthodontic force application,positive at 5 d and weak positive at 7 and 14 d.Conclusion:HSP 70 is expressed in periodontal ligment at different levels in different stages during orthodontic tooth movement.

Objective To evaluate the effects of small dose of dopamine on the renal blood flow in the elderly patients undergoing cardiac valve replacement under cardiopulmonary bypass (CPB).Methods Sixty elderly patients,of ASA physical status Ⅱ or Ⅲ,aged 65-74 yr,weighing 52-77 kg,scheduled for elective cardiac valve replacement under CPB,were randomized to receive either normal saline (group C,n =30) or dopamine (group D,n =30).After beginning of surgery,CPB was established routinely.In group D,dopamine was continuously infused for 20 min at a rate of 2 μg· kg-1 · min-1 starting from 10 min after the hearts were perfused with cardioplegic solution for the first time,while the equal volume of normal saline was given in group C.The left renal blood flow velocity was measured by transesophageal echocardiography and mean arterial pressure was recorded before and after dopamine infusion.Blood samples were obtained before surgery and at 24 h after surgery for determination of blood urea nitrogen concentrations.Results Blood urea nitrogen concentrations were significantly increased at 24 h after surgery than that before surgery in the two groups.There was no significant difference in mean arterial pressure and the left renal blood flow velocity before and after dopamine infusion between the two groups.Conclusion Small dose of dopamine (2 μg· kg-1· min-1) dose not increase the renal blood flow or improve the postoperative renal function in the elderly patients undergoing cardiac valve replacement under CPB.

Visfatin was recently reported as an adipokine and was found to exert insulin-mimicking effects.The results showed that the expression of visfatin parallelled with obesity and insulin resistance in long term high fat chow-fed rats.The expression of visfatin mRNA was decreased and the insulin resistance improved after rennin-angiotensin system was blocked.Visfatin may play an important role in the pathogenesis of insulin resistance.

Adenoma ; metabolism ; pathology ; surgery ; Humans ; Keratin-7 ; metabolism ; Kidney Neoplasms ; metabolism ; pathology ; surgery ; Kidney Transplantation ; Living Donors ; Male ; Mucin-1 ; metabolism ; Young Adult

Objective To explore the effect of Sunset Abelmoschus on podocyte injury in adriamycin-induced nephropathy rats.Methods Fifty male SD rats were randomly divided into five groups:sham operation group (n=10),model group (n=10),Sunset Abelmoschus low dose group (0.5 g·kg-1· d-1 n=10),middle dose group (1.0 g· kg-1· d-1,n=10) and high dose group (2.0 g· kg-1· d-1,n=10).Unilateral nephrectomy combined repeated adriamycin injection were performed to establish adriamycininduced nephropathy models.The rats were administered with the corresponding dose of Sunset Abelmoschus during the experiment period.Urinary protein,urinary N-acetyl glucose aminotransferase (NAG),serum albumin,serum creatinine and blood lipid were measured before operation and 2,4,6,8 weeks after operation.The rats were sacrificed on week 8 for the renal histological examination,including light microscope and electron microscope.Expression of nephrin was examined by immunofluorescence assays.Results As compared to model group,urinary protein and NAG significantly decreases in Sunset Abelmoschus groups in each time point,especially in high dose group (P＜0.01),meanwhile the serum albumin increased and the disturbance of lipid metabolism was improved in Sunset Abelmoschus groups (P＜0.05).Compared with sham group,Scr increased significantly in model group and Sunset Abelmoschus groups at the 4th week.At the 8th week,Scr in high dose group was lower than that in model group (P＜0.05),and the ratio of glomerular globe sclerosis and segmental sclerosis,tubulointerstitial damage reduced in Sunset Abelmoschus groups,especially in high dose group.The podocyte damage and the extent of foot process fusion were improved in Sunset Abelmoschus groups compared with model group.Expression of nephrin increased in Sunset Abelmoschus groups than that in model group.Conclusion Sunset Abelmoschus can ameliorate proteinuria and renal tissue damage of adriamycin-induced nephropathy rats,whose mechanism may be associated with the improvement of podocyte injury.

In this study, the expression of HSP70 during experimental tooth movement was dynamically observed and the relationship between HSP70 and orthodontic periodontal tissue remodeling were observed. The orthodontic appliance was placed between the right maxillary first molar and maxillary central incisors of adult SD rats to establish a rat molar movement model. Immunohistochemistry was performed 1, 3, 5, 7 and 14 day(s) after orthodontic force application to observe the expression and localization of inducible HSP70. The expression of HSP70 was strongly positive in the early stage of the tooth movement, became gradually less positive, and was weakly positive in the restoration stage. There was difference in staining pattern between different parts of PDL during the same period. These results suggest that the expression of HSP70 and difference in staining pattern among different parts of PDL during orthodontic tooth movement in rats may be implicated in stress response and remodeling of periodontal tissue.

Objective To extract the Ginseng polysaccharide (GPS), polysaccharides of Tricholoma matsutake (PTM)and polysaccharide of Lentinus edodes (PLE)from gingeng, tricholoma matsutake and lentinus edodes respectively,and to analyze and identify their structures,and to prepare their complex,and to study the indirect antitumor activity invitro of polysaccharide complex.Methods The polysaccharides were extracted with hot water and precipitated by ethanol.The carbohydrate levels were determined by the method of phenol-sulfuric acid.The m-hydroxyphenyl method was used to determine the levels of uronic acid, and the national standard method was used to determine the levels of starch.Infrared spectroscope and chemical methods were performed to analyze their structures. Orthogonal experiment was used to study mixing methods. Cytotoxic T lymphocyte experiment and LDH release assay were performed to detect the influence of polysaccharide complex of GPS,PTM,and PLE in the CTL killing activity,and its indirect killing effect on the P815 cells.Results The extraction rates of GPS,PTM, and PLE were 8.85%,9.40%,and 10.50%;the levels of total polysaccharides were 62.96%,59.13%,and 33.86%;the levels of uronic acid were 16.44%,9.37%,and 16.44%;the starch levels were 7.26%,2.80%,and 3.77%,respectively.The identification results showed that the polysaccharides were obstrained.When the quality ratio of the three kinds of polysaccharides was 1∶1∶1 and the concentration was 600 mg·L-1 ,the CTL cytotoxicity was the highest.Conclusion The polysaccharide complex is obtained,identified and characterized. Polysaccharide complex can enhance the cytotoxicity of CTL and has the indirectly inhibitory effect on the proliferation of P815 cells.

We aimed to study the effect of allitridum (All) on the transient outward potassium current (Ito) of ventricular myocytes of spontaneously hypertensive rats (SHR). Totally 30 male SHRs were randomly divided into three groups: low-dose All group (7.5 mg·kg(-1)), high-dose All group (15.0 mg·kg(-1)) and normal saline group. The other 10 sex and age matched Wistar-kyoto rats (WKY) were also taken as control group (WKY group). All animals received i.p. administration for 8 weeks. The dual enzymatic method was used to separate single ventricular myocyte from animals. Patch-clamp technique was used to record Ito and analyze the effect of All on the current. It was shown that the left ventricular hypertrophy of SHR was reversed significantly by All. Furthermore, the density of Ito was recovered in both high and low dose All groups. The peak current densities of Ito were enhanced from 18.23±3.64 to 25.17±2.86 pA/pF (P<0.01) and 36.47±5.42 pA/pF (P<0.01) at +50 mV by All 7.5 mg·kg(-1) and 15.0 mg·kg(-1), respectively, which was not significantly different with WKY group. The effect was associated with positive shift of the steady-state, close-state inactivation, and shortened recovery from inactivation of Ito. It is concluded that All decreases the remodeling of Ito of ventricular hypertrophic myocytes of SHR.
Allyl Compounds ; pharmacology ; Animals ; Hypertrophy, Left Ventricular ; drug therapy ; Male ; Myocytes, Cardiac ; cytology ; drug effects ; Patch-Clamp Techniques ; Potassium Channels ; metabolism ; Rats ; Rats, Inbred SHR ; Rats, Inbred WKY ; Sulfides ; pharmacology

Objective To investigate the effects of recombinant human fibronectin fragment (RetroNectin) combined with anti-CD3 monoclonal antibody (CD3Ab) on the proliferation and cytotoxicity of cytokine-induced killer cells (CIK) from acute leukemia (AL). Methods Mononuclear cells (MNCs) were isolated from peripheral blood of complete remission AL pa-tients. The MNCs were cultured in vitro by precoating with RetroNectin (RN group), CD3Ab (CD3Ab group), RetroNectin com-bined with CD3Ab (RN+CD3Ab group) and traditional method (control group) to generate CIK. The changes of growth rate, characterization, cytotoxicity and apoptosis of CIK were determined between groups. Results The amplification of CIK was higher in experimental group than that of control group, and the amplification of CIK was higher in group RN+CD3Ab than that of in group RN and group CD3Ab (P＜0.05). The expression of CD25 positive cells was higher in group RN and group RN+CD3Ab than that of group CD3Ab and control group (P＜0.05).The percentage of G1 stage cells was lower in group RN and group RN+CD3Ab than that of group CD3Ab and control group. The percentage of S stage cells was higher in group RN and group RN+CD3Ab than that of group CD3Ab and control group (P＜0.05). The cytotoxicity was higher in group RN and group RN+CD3Ab than that of group CD3Ab and control group (P＜0.05) at the E/T scope 40∶1.The percentage of apoptotic cells was lower in group RN and group RN+CD3Ab than that of group CD3Ab and control group (P ＜ 0.05). Conclusion These in vitro studies suggest that a higher activity of immune cells could be obtained by CIK cells cultured by precoating Ret-roNectin and CD3Ab.