Objective:To investigate the influence of isoflurane on neural stem cells and its possible mechanism .Methods:Mouse neural stem cells ,isolated and cultured in vitro ,were divided into the control group ,the phosphate‐buffered saline (PBS) group and the isoflurane group .The control group was not intervened .The isoflurane group exposed to isoflurane for 24 h ,while the PBS group exposed to the same amount of PBS .Then CCK‐8 assay was used to detect the cell proliferation rate . Real‐time quantitative polymerase chaln reaction was used to detect the mRNA levels of apoptosis‐related genes .Expression of apoptosis‐related protein was measured with Western blotting .Annexin V‐FITC/PI kit was used to assess cell apoptosis . Results:Compared with those in the control group and the PBS group ,cell proliferation rate ,as well as the mRNA and protein levels of Notch‐1 ,CBF‐1 and Hes‐1 ,in the isoflurane group ,decreased significantly .There were more apoptotic cells in the isoflurane group than in the control group and the PBS group .Conclusions:Isoflurane can induce the apoptosis of mouse neural stem cells ,and the process is closely related to the inhibition of Notch‐1 pathway .

Objective To investigate the effect of gabapentin on the activation of glial cells in the spinal cord after chronic constrictive injury (CCI) to sciatic nerve in rats.Methods Twenty-four male SD rats weighing 180-220 g were randomly divided into 3 groups (n = 8 each): group Ⅰ sham operation (group S), group Ⅱ CCI and group Ⅲ gabapentin + CCI. Right sciatic nerve was exposed and 4 loose ligatures were placed with 6-0chromic catgut. Seven days after operation gabapentin 50 mg/kg in 5 ml was given by intragastric gavage twice a day for 5 days in group Ⅲ. Paw withdrawal threshold to mechanical stimulation with von Frey filaments was measured one day before (baseline) and at 7, 15 d after operation. The animals were killed at 15 d after operation. The lumbar segment L4-5 of the spinal cord was removed. Immunohistochemical double mark technique was used to detect the activation of astrocytes and microglias in the spinal cord. Results Paw withdrawal threshold to mechanical stimulation was significantly decreased on the 7th and 15th day after CCI operation in group CCI as compared with group S. After 5 day treatment with gabapentin, the withdrawal threshold to von Frey hair stimulation was significantly higher in group Ⅲ than in group Ⅱ . The activation of astrocytes and microglias in the spinal cord was significantly enhanced in group CCI as compared with group S. Treatment with gabapentin significantly inhibited CCI-induced activation of astrocytes and microglias in the spinal cord. ConclusionGabapentin reduces neuropathic pain by inhibiting activation of glial cells in the spinal cord.

Objective To investigate The effect of Heat shock protein 70(HSP70) antisense oligonucleotides (ASO)to bladder carcinoma in mouse loaded with tumor.Methods The 40 mice loaded with tumor subcutaneously were established by cultured BIU-87 cells,and divided into 4 groups randomly when the subcutaneous neoplasms grew to about 100 mm3,namely,HSP70 mRNA ASO plus mitomycin C(MMC)group;HSP70 mRNA ASO group;MMC and blank control.HSP70 mRNA ASO were injected into neoplasms,10mmg/kg weight,twice every week,and MMC 0.1mg/kg weight,twice every week,and the above schemes were replaced with normal saline to blank.The neoplasms were peeled off,photograghed and weighed in 30 days.HSP70 expressions were examined with reverse transcription polymerase chain reaction (RT-PCR),mierovaseular density(MVD)was evaluated by immunohis to chemical staining and the tumor cells apoptosis was detected by terrainal deoxynucleotidyl transferase(TdT)-mediated dUTP-biotin nick end labeling technique (TUNEL).Results The tumor inhibition rate in ASO+MMC surpassed 50%.more than ASO or MMC respectively,and the differences were significantly(P＜0.05).The ASO and MMC exceeded blank group respectively(P＜0.05).The ASO was the same as the MMC(P＞0.05).The apoptotic index(AI)in ASO+MMC surpassed the other three groups (P＜0.05).The difference between ASO and MMC was not significant (P＞0.05),while the A1 of ASO or MMC was more than blank respectively(P＜0.05).The results of MVD were in accordance with the above results.Conclusion The injection of HSP70 mRNA ASO in tumor locally can inhibit neoplasm growth,and this effect might correlate with the inhibition of apoptosis and microvascular forming resulting from the ASO.

Objective To investigate the inhibitive effect of siRNA(small interfering RNA,siRNA) on the phosphodiesterase type 5(PDE5) in smooth muscle cells of human corpus cavernosum,and provide experimental groundwork for the gene therapy of erectile dysfunction (ED). Methods Small interfering RNAs targeting PDE5 gene were systhesized by using web design software provided by Ambion,there siRNAs and control siRNA were systhesized by Ambion. SiRNAs were transfected into smooth muscle cells of human corpus cavernosum by using siPORTTM Lipid reagent;down-regulation of PDE5 mRNA was detected by RT-PCR;the inhibitive effect of PDE5 was detected by Western Blotting. Results The results of RT-PCR indicated siRNA1、siRNA2 and siRNA3 made down-regulations of PDE5 mRNA expression in the transfected groups 58.2%、14.9% and 11.8%;the PDE5 expression decreased 70.5%、19.8% and 17.3%;however the expression did not have different in control siRNA and frank group. Conclusions The synthesized siRNAs in vitro were able to down-regulate the expression of PDE5.There were different capabilities of the specific siRNAs down-regulation.It was suggested that the siRNA technique provide not only an extremely powerful tool for the functional analysis of genome but also a new method for ED gene therapy.

Objective To investigate the effect of cyclooxygenase-2(COX-2) inhibitor celecoxib on bladder cancer xenografts in nude mice and apoptosis of tumor cells in the xenografts. Methods Models of bladder cancer xenograft in nude mice was used to observe the effect of celecoxib on the animals and the xenografts.TUNEL was used to assess apoptotic index of tumor cells in the xenografts. Results Celecoxib could effectively inhibit the growth of xenografts(P0.05). Conclusions Celecoxib maybe inhibit the growth of bladder cancer via inducing apoptosis of tumor cells and perhaps will become a choice of chemoprevention and adjuvant therapy of bladder cancer.

OBJECTIVETo analyze the effect of castration on risk factors for arteriosclerosis of patients with prostate cancer.

METHODSThirty patients with primary regional prostate adenocarcinoma limited to the prostate theca were selected in this study. Serum levels of testosterone (T), free testosterone (FT), dehydroepiandrosterone (DHEA), sex hormone-binding globulin (SHBG), prostatic specific antigen (PSA), triglyceride (TG), total cholesterol (TC), high density lipoprotein-cholesterol (HDL-C), low density lipoprotein-cholesterol (LDL-C), apoprotein alpha(1) (APOalpha(1)) and apoprotein beta (APObeta), insulin, plasma fibrinopeptide A (FPA), plasminogen activator inhibitor-1 (PAI-1) and fibrinogen were determined just prior to, 1 week and 1, 4 and 8 months after castration.

RESULTST, FT and PSA decreased significantly 1 week after castration (21.12 +/- 15.11 ng/ml vs 383.9 +/- 62.6 ng/ml, P < 0.001; 4.08 +/- 3.29 pmol/L vs 34.11 +/- 11.59 pmol/L, P < 0.001; 14.34 +/- 7.77 ng/ml vs 23.51 +/- 6.57 ng/ml, P = 0.001, respectively) and continued to decrease until reaching their lowest levels 8 months after castration. DHEA and SHBG did not undergo any changes. TG, fasting insulin and glucose, 2-hour insulin and glucose levels were significantly elevated 1 month after castration (1.84 +/- 0.61 mmol/L vs 1.30 +/- 0.40 mmol/L, P < 0.05; 18.16 +/- 5.57 mU/L vs 9.47 +/- 3.81 mU/L, P < 0.05; 4.77 +/- 0.66 mmol/L vs 3.92 +/- 0.34 mmol/L, P < 0.05; 65.52 +/- 14.78 mU/L vs 36.94 +/- 17.12 mU/L, P < 0.01; 6.98 +/- 0.79 mmol/L vs 6.01 +/- 0.23 mmol/L, P = 0.001, respectively). TC, LDL-C, FPA and PAI-1 levels were elevated 4 months after castration (6.56 +/- 0.99 mmol/L vs 5.29 +/- 0.75 mmol/L, P < 0.01; 4.09 +/- 0.86 mmol/L vs 3.04 +/- 0.15 mmol/L, P < 0.01; 3.39 +/- 1.67 nmol/L vs 1.48 +/- 0.50 nmol/L, P < 0.01; 27.02 +/- 5.98 ng/ml vs 21.78 +/- 3.16 ng/ml, P < 0.05, respectively), continuing to increase after that point. Insulin sensitive index (ISI) decreased significantly 1 month after surgery (-4.42 +/- 0.36 vs -3.50 +/- 0.39, P < 0.001), and continued to decrease from that point forward. HDL-C, APOalpha(1), APObeta and fibrinogen remained at pre-operative levels. There was a negative linear correlation between FT and TG, TC, LDL-C, PAI-1, FPA, fasting insulin and glucose, 2-hour insulin and glucose (r = -0.311, -0.384, -0.385, -0.339, -0.353, -0.381, -0.303, -0.460 and -0.395, respectively; P < 0.05). A similar phenomenon occurred with T (r = -0.308, -0.309, -0.356, -0.320, -0.430, -0.453, -0.435, -0.483 and -0.512, respectively; P < 0.05). T and FT were positively associated with ISI (r = 0.555 and 0.501; P < 0.001).

CONCLUSIONSAt 8 months follow-up of the study subjects, we found that lower androgen levels have adverse effects on lipid metabolism, coagulative function and insulin sensitivity, related to arteriosclerosis in men.

Aged ; Arteriosclerosis ; etiology ; Humans ; Hyperinsulinism ; complications ; Insulin Resistance ; Lipids ; blood ; Male ; Middle Aged ; Orchiectomy ; adverse effects ; Prostatic Neoplasms ; blood ; surgery ; Risk Factors

OBJECTIVETo investigate a non-invasive, effective and rapid mode of detecting the recurrence of bladder cancer during follow-up.

METHODSNinety patients following transurethral resection of bladder tumor (TURBt) surgery were recruited from January 1998 to March 2000. Standard ELISA was used to determine the quantity of nuclear matrix protein (NMP-22) in urine of all bladder cancer patients during their follow-up periods. Urine bladder tumor antigen (BTA) stat test was simultaneously performed and followed by cystoscopy.

RESULTSThe total positive rates of urinary NMP-22 and BTA stat test were 76.7% (33/43) and 67.4% (29/43), respectively. Comparatively, this positive rate would increase to 93.0% (40/43) when the combination of both urine NMP-22 and BTA test were adopted.

CONCLUSIONExamination of NMP-22 in urine is a rapid and effective way to detect the recurrence of bladder cancer. If combined with BTA test, NMP-22 may be used as a non-invasive method in surveillance of recurring of bladder cancer, which may reduce the frequency of patients needing to undergo conventional invasive cystoscopy.

Antigens, Neoplasm ; analysis ; Humans ; Neoplasm Recurrence, Local ; diagnosis ; Neoplasm Staging ; Nuclear Proteins ; urine ; Sensitivity and Specificity ; Urinary Bladder Neoplasms ; diagnosis

OBJECTIVESTo detect the level of matrix metalloproteinase-2 (MMP-2) mRNA and the tissue inhibitor of metalloproteinase-2 (TIMP-2) mRNA in bladder transitional cell carcinoma (BTCC), and to estimate the prognosis for bladder tumor based on the quality and quantity of MMP-2 and TIMP-2 mRNA.

METHODSThirty-five samples of human BTCC and 15 normal fresh bladder tissues were studied by RT-PCR analysis followed by computer-assisted image analysis.

RESULTSThe level of the MMP-2 mRNA in BTCC was significantly increased compared with that in normal bladder epithelium. The positive rates of MMP-2 and TIMP-2 mRNA were 71.4% and 65.7% in BTCC, and 66.7% and 60.0% in the normal bladder wall. The expression intensity of the MMP-2 mRNA by image analysis tended to increase with tumor grading and staging, which showed statistical significance. Similarly, the MMP-2 to TIMP-2 ratio also showed statistically significant difference between normal bladder tissue and bladder carcinoma (P < 0.01).

CONCLUSIONSA high level of the MMP-2 mRNA exists in BTCC, which may function to damage collagen IV inside the basement membrane and the extracellular basement of the bladder. The level of the MMP-2 mRNA is proportional to BTCC grading and staging, which may have prognostic value. The MMP-2 to TIMP-2 ratio may play a more significant role in the determination of the aggressiveness and prognosis of bladder tumor.

Adult ; Aged ; Aged, 80 and over ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Male ; Matrix Metalloproteinase 2 ; genetics ; Middle Aged ; Neoplasm Staging ; Predictive Value of Tests ; Prognosis ; RNA, Messenger ; genetics ; metabolism ; Tissue Inhibitor of Metalloproteinase-2 ; genetics ; Urinary Bladder ; metabolism ; pathology ; Urinary Bladder Neoplasms ; genetics ; pathology

Objective　To study the effects of transforming growth factor-β1 (TGF-β1) and transforming growth factor-α (TGF-α) on the growth and invasive potentials in human bladder tumor cells.　Methods　The effects of TGF-β1 and TGF-α on the growth of EJ cell line and the effects on MMPs and TIMP-2 were studied by means of MTT,Western Blot and RT-PCR methods.　Results　(1)TGF-β1 and TGF-α tended to inhibit the growth of EJ cells but statistically nonsigficant.(2)Higher levles of MMP-9 mRNA but lower levels of MMP-2,TIMP-2,MT1-MMP mRNA were found in EJ cells following the treatment with TGF-β1.The same was true for the expression of MMP-2,TIMP-2 mRNA in TGF-α groups.However,MMP-9 mRNA was not found in both TGF-α groups and the control groups. (3)TGF-β1 (0.1,1.0 ng/ml) enhanced MMP-2 protein but not the TIMP-2 protein,while TGF-β1 (5.0,10.0 ng/ml)decreased TIMP-2 protein but not on MMP-2.In TGF-α groups,when the concentration was 1.0,5.0,10.0ng/ml,TIMP-2 protein expression was decreased but MMP-2 did not.When the concentration reached 100.0 ng/ml,it increased MMP-2 protein level,not the TIMP-2 protein.　Conclusions　TGF-β1 and TGFα do not inhibit the proliferation of EJ cells whereas the enhanced-invasiveness and metastasis may be associated with regulating the expression of MMPs and TIMP-2.

Objective To evaluate the inhibitory effect of endostatin on bladder tumor and to investigate the possible mechanism of the inhibition. Methods (1)By means of MTT method,the effects of endostatin on ECV304 and EJ cells proliferation were studied.(2)Ten nude mice were subcutaneously implanted with EJ bladder tumor cells (1?10 7/ml). After the tumor volume exceeded 100～200 mm 3,the mice were randomized into two groups.One group received recombinant human endostatin (2 mg?kg -1 ?d -1 ) whereas the other group received PBS as control.All the treatment lasted 21 days.(3) After that,the mice were sacrificed and then MMPs and TIMPs mRNA expression were measured in implanted tumor by RT PCR and Western blot method. Results (1)Endostatin inhibits ECV304 proliferation stimulated with bFGF (5 ng/ml).In addition,we report for the first time that endostatin also inhibits EJ cells proliferation.(2)The mean tumor weight of endostatin treated group (0.70?0.16 g) was significantly lower than that of control group (1.14?0.21)g, P