Objective To study the effects of genistein and adriamycin on the proliferation of human ovarian cell A2780 and growth of tumor bearing nude mice.Methods The ovarian cancer cell A2780 suspension were inoculated into nude mice (n=40 )to establish the animal models with ovarian tumor.These model rats were randomly divided into four groups,saline group and different genistein concentrations combined with adriamycin treatment group (high-dose,middle-dose and low-dose group)by intraperitoneal injection once a week.After treatment for four weeks,the ovarian tumors in all rats were peeled off and weighted for the measurement of tumor inhibition rate.The expression of c-myc in ovarian tumors were detected by immunohistochemistry.In vitro,after affected by different genistein concentrations combined with adriamycin,the expression of c-myc in A2780 cell of each group were detected by western blot.The proliferation of A2780 cell were determined by MTT method,and the invasiveness of cell by cell adhesion and transwell assay,the apoptosis rate and cell cycle were measured by flow cytometry. Results The tumor inhibition rates in middle-dose (40.26 ± 4.25)% and high-dose groups[,(44.81 ±5.74)%])were significantly lower than saline group (P<0.05).Immunohistochemistry results showed that c-myc monoclonal antibody in middle-dose and high-dose groups can significantly inhibit the activation of c-myc in ovarian carcinoma tissue.Compared with control group,the expression of c-myc in A2780 cell were significantly decreased,the proliferation and invasiveness of cell were inhibited,cells in G0/G1 phase were significantly increased(P<0.05)and in S phase were(P<0.05)decreased,and the apoptosis were induced by genistein combined with adriamycin in middle-dose and high-dose (P <0.05 ).Conclusion Genistein combined with adriamycin in certain doses could inhibit the proliferation and invasiveness of ovarian carcinoma,and induce its apoptosis.The mechanism may be related to the decreased expression of c-myc.

Objective To evaluate the clinical applications and surgical methods of combined laparoscopic common bile duct (CBD) exploration with choledochoscopy. Methods From 2006 to 2009,clinical data of 42 patients with choledocholithiasis undergoing laparoscopic common bile duct exploration were retrospectively analyzed. We applied a step-by-step electric coagulating incision technique on the CBD,the step-by-step suturing technique, and the step-by-step clamping technique with alligator forceps, and soft tube irrigating technique with suctioning by selecting the proper exploration route, improving the common bile duct incision technique and calculus removing techniques. Results Procedures were successful in all the cases. There was no conversions to open surgery, no postoperative bleeding and no operative mortality. The mean operating time was 120 minutes (ranging, 90 to 150 minutes) with minimal intraoperative blood loss ( ranging, 20 to 40 ml). Ductal stone clearance was successful in 41 out of 42 patients ( 93% ). The largest number of the common bile duct stones was 16. With the diameter of stones larger than 15 mm in 18 cases in which the biggest was 30 mm. Bile leak developed in 1 patient, retained stones found in 3 patients,including intrahepatic cholelithiasis in one case. As a result, 38 out of 42 patients underwent common bile duct exploration. 35 patients were placed on T-tubes. Four patients underwent cystic duct exploration in which 3 had primary suture of the cystic duct and 1 had drainage. There was no infection and stenosis of biliary tract in the 42 followed-up cases. Conclusions Laparoscopic common bile duct exploration with stone extraction can be performed with high efficiency, minimal morbidity and without mortality. Improving the way of operation and selecting suitable exploration can result in better clinical outcomes.

A test was conducted to examine the effect of several electron donors such as glucose, sodium acetate, Fe0, Fe0+glucose and Fe0+sodium acetate on reductive dechlorination of 2,4-dichlorophenol (2,4-DCP) through inoculating the unacclimated anaerobic mixed bacteria. The optimum condition and sus-tainability of Fe0 as electron donor was also been discussed. The results showed that, Fe0+glucose enhanced the dechlorination of contaminant effectively compared to glucose. Sodium acetate, Fe0 and Fe0+sodium acetate were all effective electron donors and Fe0 was the optimum, the optimum initial pH was 8.0 and quantity of added Fe0 was 2.0 g/L. 4-CP was the mainly intermediate product for 2,4-DCP dechlorination. Fe0 could support the electron for reductive dechlorination of 2,4-DCP continuously. In contrast, when so-dium acetate as electron donor, the effect of dechlorination was inferior to Fe0 with the consumption of sodium acetate.

Objective To investigate the expression of musashi-1(msi-1)and its significances in small intestinal mucous severely damaged by high close 5-FU.Methods Total 40 adult C57BL/6J mice were divided into control group(n= 8,group D)and experimental group(n=32).Mice in control group received intraperitoneal injection of PBS as control,and mice in experimental group were intraperitoneally injected high dose 5-FU(150 mg per kg of body weigh for five days).After treatment 1 day(group A),3 days(group B)and 5 days( group C),the dying mice were killed,HE straining and immunohistochemical technique were carried out for detecting the expression of putative marker of intestinal epithelial stem cells——musashi-1(msi-1)in the samples of the meddle intestine,and the percentage of the msi-1 positive cells from the intestinal mucosal cells of the mice in group A was detected by flow cytometry.Results After the treatment of high dose 5-FU,the intestinal mucous was damaged severely;the number of msi-1 positive cells increased markedly;The intestinal mucosal cells can be divided into two groups by flow cytometry,and in the group in which the value of FSC was higher,the percentage of msi-1 positive cells increased to 67.75 %.Conclusions After the treatment of high dose of 5-FU,the percentage of intestinal stem cells increased significantly;this model may be useful for further isolation and enrichment of intestinal epithelial stem cells.

Objective To investigate the protective effect of basic fibroblast growth factor(bFGF) gene modified mesenchymal stem cells(MSCs-bFGF)on cerebral isehemia in rats.Methods MSCs or MSCs-bFGF were transplanted into rat models of focal cerebral ischemia by intravenous injection.The neurological deficits and infarction volumes were evaluated,and the survival rate and differentiation of grafted MSCs were observed by double immunofiuoreseent labeling.Results In the rat cerebral ischemic model, both MSCs and MSCs-bFGF showed protective effect on the rats in comparison with control group.However, the protective effect was more significant in MSCs-bFGF group.Double immunofluorescent staining showed the number of BrdU-labeled and NeuN co-expression cells in MSCs-bFGF treated animals(127.40?7.43 and 11.20?3.09)were much more than in those of MSCs treated animals.While there was no significant difference between MSCs-bFGF and MSCs group in the number of GFAP co-expression cells.Conclusion MSCs transplantation has protective effect on cerebral ischemia in rats.Basic fibroblast growth factor gene modified MSCs is more effective than MSCs in neuroproteetion.

Objective To evaluate the clinical value of transoesophageal echocardiography on perventricular closure of ventricular septal defects(VSD).Methods Transoesophageal echocardiography (TEE)was performed in 27 patients with VSD diagnosed by the transthoracic echocardiography examination (TTE)and prepared to receive perventricular closure of VSD for the purposes of choosing the appropriate occluder before VSD closure and guiding occluder release during the operation and evaluating the post operative effects of VSD closure 1 week post operation.Results Three patients with VSD not suitable for perventrieular VSD closure according to TEE results were not operated,perventricular closure of VSD were successfully performed in 20 patients and minimal residual shunt was seen in 1 patient post operation,perventricular closure of VSD failed in 4 patients and these patients received surgical closure.TEE at 1 week post successfully with perventricular closure of VSD in the 20 patients showed that there were no residual shunts and the pulmonary artery pressure was significantly lower post operation compared to that of preoperation value.Conclusion TEE is helpful in choosing the suitable patients and occlude size,guiding occlude release and evaluating the post operative results for perventricular closure of ventricular septal defects.

Objective: Cell cycle has recently become more appealing as a new target of anti-carcinogen-ic agent. Diallyl disulfide (DADS) inhibits growth and induces call cycle G_2/M arrest in human gastric cancer BGC823 cells. Cell division cycle protein 25C (Cdc25C) and CyclinB1 expression are involved in G_2/M arrest.However, mechanisms of G_2/M arrest are not yet fully understood. The aim of this study was to elucidate the mechanism of cell cycle G_2/M arrest in human gastric cancer BGC823 cells induced by DADS. Methods: The expression of chk1 and Chk2 mRNA associated with cell cycle arrest of BGC823 cells after the induction with DADS for 1 or 2 days was detected by RT-PCR. The protein expression of cycle-related proteins ATM-RAD3-related gene (ATR), checkpoint kinase1 (Chk1), checkpoint kinase 2 (Chk2), P-ATR, P-Chk1 and P-Chk2 was measured by Western blot. Interaction between Chk1/2 and Cdc25C was analyzed by immuno-precipitation. Results: After the cells were treated with 15 mg/L DADS for 1 or 2 days, the expression of Chk1 and Chk2 mRNA was not significantly different from that in untreated cells (P>0.05). Western blot analysis showed that the expression of total Chk1 and Chk2 treated with 15 mg/L DADS was not significantly different from that in untreated cells. But phospho-chk1 showed a significant increase after stimulation with 15 mg/L DADS for 2h to 12h and continued to increase gradually as time went on (P<0.05). Phospho-Chk2 showed a eak expression and a weaker expression after stimulation with DADS, but the changes were not statistically significant (P>0.05). Addition of 15 mg/L DADS to BGC823 cells for 15 rain to 120 min resulted in an increase in phospho-ATR expression, whereas no changes were found in ATR expression (P<0.05). The Chk1 Ab in-creasingly precipitated Cdc25C in BGC823 cells treated with DADS (P<0.05). In contrast, Chk2 Ab failed to change precipitation with Cdc25C by DADS (P>0.05). Conclusion: Activation of chk1 was involved in cell cy-cle G_2/M arrest in BGC823 cells treated with DADS. Cell cycle G_2/M arrest by DADS is associated with phos-phorylation of several cell cycle regulatory proteins including ATR and Chk1 which regulate expression of Cdc25C.

AIMTo investigate the change of ERK, GDNF expression activity in temple and frontal lobe of AD rat after 96 dB wide band noise exposure.

METHODSExperimental group SD rats(weight from 150-220 g), either male or female, were randomly assigned to three groups: control (n=10); physiological brine-injected control (n=8); glutamic acid-injected group (n=8) after getting rid of memory loss(indicating "a little memory" and "no memory") rats culled by behavior training. The rats were micro-injected at the stereotaxic device (AP3.2-3.4, L2.0-2.4,H2.8-3.0)by glutamic acid or same volume of physiological brine in the each side of hippocampus CA1. Western Blot and image quantitative analysis technique, combined with auditory brainstem response (ABR) measurement.

RESULTS(1) Expressions of ERK in frontal cortex for control rat are much more than that of other group, which has a obvious up-regulation after being given 96 dB noise. Expression of ERK in temple cortex for 3 groups rat have a increase(plus noise), moreover, they are stronger than expression in frontal cortex for different group. (3) GDNF in frontal cortex for control rat have a higher expression than that of the same group before adding noise (up-regulation). (4) GDNF expression in temple lobe for glutamatic acid group have a remarkable down-regulation trend. (5) Expression of GDNF in frontal for control rat is much less than in temple.

CONCLUSIONERK in frontal cortex for AD model rat have fewer expressions and not being affected by 96 dB noise, but it is reverse in temple. Wide band frequency noise can reduce the expression of GDNF in temple lobe of AD model rat.

Acoustic Stimulation ; Alzheimer Disease ; metabolism ; physiopathology ; Animals ; Auditory Threshold ; Disease Models, Animal ; Evoked Potentials, Auditory, Brain Stem ; Extracellular Signal-Regulated MAP Kinases ; metabolism ; Female ; Glial Cell Line-Derived Neurotrophic Factor ; metabolism ; Glutamic Acid ; adverse effects ; Male ; Noise ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate relationship between expressions of hypoxia inducible factor-1alpha (HIF-1alpha) and insulin-like growth factor-II (IGF-II) as well as their correlation with angiogenesis and prognosis in gastric carcinoma.

METHODSHIF-1alpha and IGF-II mRNA expression were analyzed using in situ hybridization, microvessel density (MVD) was determined by anti-CD34 immunostaining in 118 cases gastric carcinoma.

RESULTSThe positive rates of HIF-1alpha mRNA and IGF-II mRNA were 49.2% and 47.4%, respectively. In stage T3-T4 cases, positive rates of HIF-1alpha and IGF-II mRNA expression, the frequencies of vessel invasion, lymph node and distant metastasis were significantly higher than those in stage T1-T2 cases. The mean MVD in stage T3-T4 tumors, vessel invasion, lymph node metastasis and distant metastasis were significantly more frequent than those in stage T1-T2 tumors. The mean MVD in tumors with positive HIF-1alpha and IGF-II mRNA expression was significantly higher than those in tumors without HIF-1alpha and IGF-II mRNA expression. The expression of HIF-1alpha was positively correlated with IGF-II mRNA. There were positive correlations between MVD and expression of HIF-1alpha and IGF-II mRNA, respectively. The mean survival time and 5-year survival rate in cases with positive HIF-1alpha and VEGF expression and MVD value >/= 41.5 were significantly shorter than those in cases with negative HIF-1alpha and VEGF expression and MVD value < 41.5.

CONCLUSIONSHIF-1alpha and IGF-II play an important role in tumor invasion and metastasis, especially in tumor angiogenesis. So test of the expression of HIF-1alpha and IGF-II may act as a useful index of treatment and prognosis.

Adult ; Aged ; Antigens, CD34 ; analysis ; Female ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; genetics ; metabolism ; Immunohistochemistry ; In Situ Hybridization ; Insulin-Like Growth Factor II ; metabolism ; Male ; Middle Aged ; Neovascularization, Pathologic ; genetics ; metabolism ; pathology ; Prognosis ; RNA, Messenger ; genetics ; metabolism ; Stomach Neoplasms ; blood supply ; genetics ; metabolism ; Survival Analysis

OBJECTIVETo investigate the influence of total flavonoids of epimedium (TFE) on the streptozocin (STZ)-induced kidney injury in diabetic rats and discuss the possible mechanism.

METHODSDiabetes was produced by a single injection of streptozocin (40 mg/kg, iv) in male SD rats. The rats were randomly divided into three groups (n = 10): control group, model group and TFE group (100 mg/kg, ig). Animals were sacrificed 12 weeks later. The level of blood glucose, blood urea nitrogen (BUN) and creatinine (Cr) as well as the renal index were determined. Detect the specific biochemical of renal tissue: superoxide dismutase (SOD), malondialdehyde (MDA). Use masson staining to observe the morphology of the renal tissue. Immunohistochemistry was employed to determine the protein levels of transforming growth factor-beta1 (TGF-beta1).

RESULTSCompared to control group, the enhancement of blood glucose, renal index, BUN and Cr was found in model group, which was significantly attenuated by treatment with TFE. Meanwhile, elevated MDA level in renal tissue as well as decreased SOD activities in renal tissue were significantly remitted by TFE. Furthermore, TFE decreased the expression of TGF-beta1.

CONCLUSIONTFE can evidently relieve renal damage in rats with diabetic nephropathy induced by STZ, which might be related to antioxidation and modulating the expression of TGF-beta1 protein.

Animals ; Diabetes Mellitus, Experimental ; metabolism ; Diabetic Nephropathies ; metabolism ; prevention & control ; Epimedium ; chemistry ; Flavonoids ; pharmacology ; Kidney ; drug effects ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley