Objective To investigate the effect of Niaochangshu capsule on urodynamics in bladder filling period of patients with post-menopause urethral syndrome of renal deficiency and blood stasis type.Methods A total of 36 patients with post-menopause urethral syndrome of renal deficiency and blood stasis type were randomly assigned to two groups:the trial group(24 patients were treated with Niaochangshu capsule) and the control group(12 patients were treated with Nilestriol).All patients were treated for 8 weeks.The changes of the urodynamic indexes in bladder filling period(the capacity of the bladder,the compliance of the bladder and the sensitivity of the bladder) were compared between the two groups.Results After treatment,the urodynamic index,including the maximum cystic capacity,the volume at the first desire to void and the cystic compliance were evidently improved in both of the trial group and the control group(P0.05).Conclusion Niaochangshu capsule is effective in treating the patients with post-menopause urethral syndrome of renal deficiency and blood stasis type.It can increase the cystic capacity and compliance,step down the sensitivity of the bladder.Its effect is similar to Nilestriol.

Objective To investigate the changes of serum insulin-like growth factor-2 (IGF-2) and insulin-link growth factor binding protein-3(IGFBP-3) in the patients with ovarian cancer before and after operation,and evaluate their clinical significance.MethodsThe contents of serum IGF-2 and IGFBP-3 in 82 patients with ovarian cancer were detected by ELISA before and after operation and compared with that in health controls.ResultsThe contents of IGF-2 in the patients before operation were significantly higher than that in control group[(101.5±22.2)ng/ml,(49.3±15.6)ng/ml vs (69.6±17.7)ng/ml,(23.9±11.3)ng/ml,t＝3.74,2.85,P＜0.05].The contents of IGFBP-3[(39.8±11.1)ng/ml]in the patients before operation were significantly lower than that in control group[(55.8±19.2)ng/ml](t′＝4.49,P＜0.05).There was significant correlation between the contents with lymph node metastasis and clinical stage[(107.5±24.0)ng/ml,(41.7±16.9)ng/ml vs (91.6±17.7)ng/ml,(56.9±19.1)ng/ml;(103.4±27.2)ng/ml,(50.2±16.6)ng/ml vs (86.6±12.3)ng/ml,(41.1±17.1)ng/ml,t＝2.83,2.37,2.48,3.32,P＜0.05).The contents of IGF-2 was significantly decreased,and IGFBP-3 was significantly increased in patients after radical operation [(86.6±12.3)ng/ml,(41.1±17.1)ng/ml vs (103.2±26.0)ng/ml,(45.3±14.9)ng/ml,t′＝3.46,t＝2.67,P＜0.05].But there was no significant difference on the level of IGF-2 and IGFBP-3 before and after palliative resection(P＞0.05).ConclusionsThe contents of serum IGF-2 and IGFBP-3 are closely related to tumor invasion,metastasis and clinical stage.Dynamic determination of the contents of serum IGF-2 and IGFBP-3 may be an important index for evaluation of invasion,metastasis,efficacy and prognosis for the patients with ovarian cancer.

Objective To investigate the effect of downregulation of KIAA0101 protein expression on the proliferation and invasion of a cutaneous squamous cell carcinoma cell line SCL-1,and to explore possible molecular mechanisms underlying the effect.Methods SCL-1 cells were classified into three groups: siRNA control group transfected with the control siRNA,KIAA0101 group transfected with KIAA0101 siRNA,and untreated group remaining untreated.After additional culture,Western blot was used to detect the expression of KIAA0101 protein and proteins associated with cell proliferation and invasion,cell counting kit-8 (CCK-8) to evaluate cellular proliferative activity,and Boyden chamber assay to estimate invasive ability of cells.Results The relative expression level of KIAA0101 protein was 0.062 ± 0.095 in the KIAA0101 group,significantly lower than that in the untreated group (0.359 ± 0.044,P ＜0.05) and siRNA control group (0.379 ± 0.025,P ＜0.05).A significant decrease was observed in cellular proliferative activity (from 24 to 96 hours) and invasive activity (at 48 hours) in the KIAA0101 group compared with the other two groups (all P ＜0.05).Moreover,compared with the untreated group and siRNA control group,the KIAA0101 group showed a stronger expression of p21 protein (0.570 ± 0.060 vs.0.048 ± 0.018 and 0.055 ± 0.014,P ＜0.01) but a weaker expression of matrix metalloproteinase 2 (MMP2) protein (0.051 ± 0.013 vs.0.205 ± 0.029 and 0.221 ± 0.029,P ＜0.01).Conclusion The inhibition of SCL-1 cell proliferation and invasion induced by the downregulation of KIAA0101 gene expression may be associated with the expression changes of p21 and MMP2.

Objective To study the effect of down-regulation of PTTG on the proliferation and migration of cutaneous squamous cell carcinoma cell line SCL-1 and its related mechanism. Methods SCL-1 cells were transfected with control siRNA or PTTG-targeting siRNA (PTTG-siRNA), or remained untransfected. After additional culture, the proliferation of SCL-1 cells as observed with cell counting kit-8 (CCK-8), and cell migration with Boyden chamber. Real-time PCR and Western blot were performed to detect the expression of matrix metalloproteinase 2 (MMP-2), MMP-9 and PTTG. Results The proliferation of SCL-1 cells transfected with PTTG-siRNA was markedly deccelarated in comparision with that of untransfected cells and those transfected with control siRNA (both P< 0.05). Real-time PCR and Western blot disclosed a significant decrease in the mRNA and protein expression of MMP-2, MMP-9 and PTTG in PTTG-siRNA-transfected SCL-1 cells compared with the other two groups of cells. As real-time PCR showed, the expressions of MMP-2, MMP-9 and PTTG in PTTG-siRNA-transfected SCL-1 cells were 0.8%, 23.2% and 21.3% of those in untransfected cells, respectively. Further more, the number of SCL-1 cells migrating through microporous membrane in the Boyden chamber was significantly smaller in PTTG-siRNA-transfected group than in untransfected group and control siRNA-trans-fected group (51.38 ± 4.71 vs 131.33 ± 6.12 and 127.72 ± 5.20, both P< 0.05). Conclusion The down-regulation of PTTG may deccelarate the proliferation and migration of SCL-1 cells and inhibit the expression of MMP-2 and MMP-9 in SCL-1 cells.

Apoptosis ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Cell Line, Tumor ; Down-Regulation ; Glucosephosphate Dehydrogenase ; metabolism ; Humans ; Neoplasm Invasiveness ; Skin Neoplasms ; metabolism ; pathology

ObjectiveTo investigate the role of Notch1 gene in xenografted human cutaneous squamous cell (SCL-1) carcinoma. MethodsFifteen nude mice were divided into three groups, including untreated group(inoculated with SCL-1 cells treated with phosphate buffered saline), empty vector group (inoculated with SCL-1 cells transfected with empty vector) and Notch1 group(inoculated with SCL-1 cells transfected with Notch1 expression vector). All the mice were inoculated with SCL-1 cells(1 x 108/ml) of0.2 ml. Then, the growth of xenografted tumor was observed every other day. Fifteen days later, the mice were sacrificed, tumor tissue was dissected and subjected to terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay for the detection of cell apoptosis, reverse-transcription(RT)-PCR and Western blot for the examination of mRNA and protein expressions of Notch1, bcl-2 and bax, respectively. ResultsThe proliferation of xenografted tumor in Notch1 group was obviously inhibited compared with the untreated group. The weight of xenografted tumor in Notch1 group was significantly lower than that in the untreated group and empty vector group (0.574 ± 0.219 g vs. 2.642 ± 0.404 g and 2.606 ± 0.512 g, F= 26.642, P＜ 0.01). TUNEL assay demonstrated that the number of apoptotic cells per 500 cells in tumor tissue specimens was(87 ± 9) in Notch1 group, evidently higher than that in the untreated group(8 ± 2) and empty vector group(10 ± 3) (F = 194.266, P ＜ 0.05 ). Further, RT-PCR and Western blot revealed that the mRNA and protein expressions of Notch1 and bax were significantly upregulated, but those of bcl-2 were markedly downregulated in the Notch 1 group, with significant difference among the three groups(all P ＜ 0.05). ConclusionsNotch 1 gene can inhibit the growth of xenogra ffted human cutaneous squamous cell(SCL-1) carcinoma and induce SCL-1 cell apoptosis likely by upregulating bax expression and downregulating bcl-2 expression.

Objective To investigate the correlation of heme oxygenase-1 (HO-1) expression and autophagy in mice with liver ischemia reperfusion (IR) injury,and to study the influence of HO-1 on the hepatic function.Methods Control group,IR group,Hemin + IR group and Znpp + IR group were constructed.Mice in the Hemin + IR group and the Znpp + IR group were pretreated by Hemin (HO-1 inducer) and Znpp (HO-1 inhibitor) before IR.According to different reperfusion time,the IR group was divided into the 0,2,6,12,24,48 and 72 hours groups,and the Hemin + IR group and the Znpp + IR group were divided into the 0,2,6,12,24 hours groups.The levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were detected to evaluate the hepatic function.The autophagosome and pathological changes of liver were observed under electron microscope and hematoxylin and eosin (HE) staining,respectively.The expressions of HO-1 and autophagyassociated protein (LC3Ⅱ) in the protein level were detected by Western blot.All data were analyzed using the analysis of variance,t test and q test,respectively.Results The levels of ALT and AST of the IR group,Hemin + IR group and Znpp + IR group were significantly higher than those of the control group (F =96.17,85.53,P ＜0.05).The levels of ALT and AST of the IR 12 hours group and 24 hours group were significantly higher than those of the Hemin + IR 12 hours group and 24 hours group (qALT=--14.46,-7.85 ; qAST=-12.98,-5.26,P ＜0.05),but significantly lower than those of the Znpp + IR 12 hours group and 24 hours group (qALT=4.25,4.94;qAST=4.98,3.53,P ＜ 0.05).The results of HE staining showed that hepatic IR injury was significalty alleviated in the Hemin + IR group when compared with the IR group and the Znpp + IR group.The Suzuki degrees of hepatic IR injury in the control group,IR 12 hours group,Hemin + IR 12 hours group and the Znpp + IR 12 hours group were 0.5 ± 0.3,2.6 ± 0.5,1.1 ± 0.3,3.0 ± 0.4,respectively,with significant differences among the 4 groups (F =53.62,P ＜0.05).There were significant difference in the Suzuki degrees of hepatic IR injury between the IR 12 hours group and the Hemin + IR 12 hours group,and between the IR 12 hours group and the Znpp + IR 12 hours group (q =10.67,14.02,P ＜ 0.05).The results of electron microscopy showed that the number of autophagosome in the IR group was greater than the control group and Znpp + IR group,but lesser than the Hemin + IR group.The numbers of autophagosome in every 10 high power fields of the control group,IR 12 hours group,Hemin + IR 12 hours group and the Znpp + IR 12 hours group were 0.4 ±0.2,1.8 ±0.6,4.0 ± 1.8,0.7 ±0.5,respectively,with significant difference among the 4 groups (F =21.35,P ＜ 0.05).There were significant differences in the number of autophagosome between the IR 12 hours group and the Hemin + IR 12 hours group,and between the IR 12 hours group and the Znpp + IR 12 hours group (q =6.05,-3.03,P ＜ 0.05).The expressions of HO-1 and LC3Ⅱ in the IR group and the Hemin + IR group were significantly increased.The relative expressions of HO-1 protein in the Hemin + IR 6 hours group and the 12 hours group were 0.64 ±0.17 and 0.51 ±0.12,which were significantly higher than 0.45 ± 0.08 and 0.17 ± 0.03 of the IR 6 hours group and 12 hours group (t =4.03,4.69,P ＜ 0.05).The relative expressions of LC3Ⅱ protein in the Hemin + IR 6 hours group and the 12 hours group were 1.04 ± 0.20 and 1.20 ± 0.23,which were significantly higher than 0.58 ± 0.04 and 0.95 ±0.14 of the IR 6 hours group and 12 hours group (t =4.29,6.69,P＜0.05).The relative expressions of HO-1 protein in the Znpp + IR 6 hours group and 12 hours group were 0.23 ± 0.03,0.14 ± 0.02,respectively,and the relative expressions of LC3Ⅱ protein were 0.35 ± 0.04 and 0.49 ± 0.14,which were significantly lower than the HO-1 and LC3Ⅱ protein expressions in the IR 6 hours group and 12 hours group (tHO-1 =13.82,7.04; tLC3Ⅱ =7.21,4.03,P ＜0.05).Conclusion HO-1 is positively related to the amelioration of hepatic function,and it may be achieved by induction of autophagy.

Objective To evaluate effects of downregulation of glucose?6?phosphate dehydrogenase(G6PD) expression on proliferation and cell cycle distribution of cutaneous squamous cell carcinoma(CSCC)cells. Methods Western blot analysis was performed to measure the protein expression of G6PD in normally cultured human HaCaT keratinocytes, SCL?1 and A431 CSCC cells. When A431 cells grew to 85%-90%confluence, a small interfering RNA (siRNA)targeting G6PD(G6PD?siRNA group)and a negative control siRNA(siRNA control group)were transfected into them separately, and untransfected A431 cells served as the untransfected group. CCK?8 assay was performed to evaluate proliferative activity of the A431 cells on days 0, 1, 2, 3 and 4 after transfection, Western blot analysis to measure G6PD, cyclin D1 and CDK4 protein expressions in A431 cells, and flow cytometry to analyze cell cycle distribution in A431 cells after 48 hours of additional culture. Results The protein expression of G6PD was significantly higher in normally cultured SCL?1 cells(0.308 ± 0.023)and A431 cells(0.643 ± 0.046)than in HaCaT cells(0.100 ± 0.019, both P<0.05), and significantly higher in A431 cells than in SCL?1 cells(P<0.05). The G6PD?siRNA group showed significantly decreased protein expressions of G6PD, cyclin D1 and CDK4(0.134 ± 0.027, 0.154 ± 0.017 and 0.166 ± 0.017, respectively)compared with the untransfected group(0.425 ± 0.029, 0.344 ± 0.024 and 0.330 ± 0.020 respectively)and siRNA control group(0.444 ± 0.033, 0.350 ± 0.027 and 0.348 ± 0.018 respectively) (all P<0.05). Besides, the G6PD?siRNA group showed significantly decreased cellular proliferative activity on days 1-4 compared with the siRNA control group and untransfected group(all P<0.001), while there were no significant differences between the untransfected group and siRNA control group at any of the time points (all P > 0.05). Compared with the untransfected group and siRNA control group, the G6PD?siRNA group showed significantly higher proportions of A431 cells in G0/G1 phase(both P < 0.001), but significantly lower proportions of A431 cells in S phase(both P<0.001). Conclusion G6PD may play important roles in the regulation of proliferation and cell cycle distribution of CSCC cells.

Objective To investigate the effect of pituitary tumor-transforming gene (PTTG)siRNA on the growth,invasion of,and expression of metastasis-related cytokines including matrix metalloproteinase-2 (MMP-2)and MMP-9 in xenografted human cutaneous squamous cell carcinoma in nude mice.Methods SCL-1 cells were subcutaneouslv inoculated into Balb/c nude mice to establish a xenograft model of human cutaneous squamous cell carcinoma.Then,15 mice bearing xenografted carcinoma were equally divided into 3 groups to be inoculated with phosphate buffer saline (PBS),control siRNA,and PTTG siRNA of 50 nmoI/L,respectively,ever),other day for 2 weeks.The size of xenograted carcinoma in these mice was measured every other day.At the end of 2-week treatment.the mice were killed followed by the evaluation of tumor weight,as well as the quantification of mRNA and protein expression of PTTG,MMP-2 and MMP-9 by reverse transcription (RT)-PCR and Western-blot,respectively.Results The xenograft model of human cutaneous squamous cell carcinoma was successfully established.The treatment with PTTG siRNA obviously inhibited the growth of the xenografted tumom and the expression of PTTG mRNA and protein compared with PBS and control siRNA (all P<0.05).In addition,the expression of MMP-2 and MMP-9 in xenografted tumors in PTTG siRNAtreated mice were significantly lower than those in PBS and control siRNA-treated mice.suggesting that PTTG siRNA evoked the decrease in invasive and metastatic ability of xenografted tumors.Conclusions PTTG siRNA can inhibit the growth of human cutaneous squamous cell carcinoma xenografts in nude mice,and downregulate the expression of invasion-and metastasis-related cytokines,including MMP-2 and MMP-9.

ObjectiveTo investigate the role and clinical pathological significance of PTTG and bFGF in cutaneous squamous cell carcinoma(CSCC).MethodsTissue specimens were collected from the lesions of 42 patients with CSCC and normal skin of 42 normal human controls.The protein and mRNA expressions of PTTG and bFGF were detected by immunohistochemistry and in situ hybridization in these specimens respectively.ResultsA significant increase was observed in the positive expression rates of PTTG and bFGF proteins[64.3％(27/42) vs.11.9％(5/42),73.8％(31/42) vs.21.4％(9/42),both P＜ 0.05] and mRNA [59.5％(25/42) vs.7.1％(3/42),75.0％(29/42) vs.16.7％(7/42),both P＜ 0.05] in the CSCC tissue specimens than in the control specimens.The protein and mRNA expressions of PTTG were positively correlated with those of bFGF(both P ＜ 0.05),and closely correlated with histological grade of CSCC (both P ＜0.05).ConclusionThe high expression of PTTG and bFGF may be associated with the initiation of CSCC.