Mechanisms of resveratrol bovine serum albumin nanoparticle-induced cell death in human ovarian cancer SKOV3 cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the effect of resveratrol bovine serum albumin nanoparticles on SKOV3 cell line and its mechanisms.</p><p><b>METHODS</b>The morphological changes of the cells exposed to the nanoparticles were observed by apoptotic body/cell nucleus DNA staining under inverted microscope and fluorescence microscope, and the pathway of cell death was determined by phosphatidylserine translocation. Western blotting was performed to detect the activation of cyto.c, caspase-3 and caspase-9.</p><p><b>RESULTS</b>DNA ladder was detected with gel electrophoresis and the cell death was partially inhibited by the pan-caspase inhibitor Z-VAD-FMK. Gel electrophoresis displayed both DNA ladder and smear in RES-BSANP exposed groups, while DNA ladder disappeared in Z-VAD-FMK group and only the smear was left. Cyto.c in the cytoplasm was released at 2 h, while the expression of caspase-9 protein reached the peak level at 4 h and caspase-3 expression was obvious enhanced at 8 h. At 4 h, caspase-9 expression in the cells exposed to 100 µmol/L RES-BSANP was decreased significantly as compared to the cells treated with 50 µmol/L RES-BSANP (P>0.05).</p><p><b>CONCLUSION</b>RES-BSANP can induce the necrosis and apoptosis of SKOV3 cells via either caspase-dependent or caspase-independent pathways.</p>

Effects of yikunning on the expressions of bcl-2 and bax in rat ovaries during perimenopausal period / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To investigate the effects of Yikunning (YKN, Chinese Traditional Medicine) on the expressions of bcl-2 and bax in rat ovaries during perimenopausal period.</p><p><b>METHODS</b>Thirty female Wistar rats during perimenopausal period were selected by unforced aging. Then the rats were divided into 3 groups randomly: YKN group, Livial control group and Aged control group. Ten young female Wistar rats were selected as young control group. Intragastric administrations were conducted for 4 weeks once daily continuously. The expressions of Bcl-2 Bax mRNAs and proteins in rat ovaries were detected by RT-PCR and Western blot.</p><p><b>RESULTS</b>The levels of Bcl-2, Bax mRNAs and proteins in rat ovaries in YKN group were higher than those in Aged control group, which showed differences among them (P < 0.01). The Bcl-2/Bax mRNA rate and protein rate in rat ovaries in YKN group were higher than those in Aged control group, which showed differences among them (P < 0.05 or P < 0.01).</p><p><b>CONCLUSION</b>YKN could increase the expressions of Bcl-2, Bax mRNAs and proteins and up-regulate the Bcl-2/Bax mRNA rate, protein rate in rat ovaries during perimenopausal period, which might be one of the molecular mechanisms of YKN postponed the ovarian failure and cured perimenopausal syndrome.</p>

Effects of Yikunning on the apoptotic rate and expression of caspase-3 in rat ovaries during perimenopausal period / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To investigate the effects of Yikunning (compound of Chinese traditional Medicine, YKN) on the apoptotic rate and expression of caspase-3 in rat ovaries during perimenopausal period.</p><p><b>METHODS</b>Thirty female Wistar rats during perimenopausal period were selected by unforced aging. Then the rats were divided into 3 groups randomly: YKN group, livial control group and aged control group. Ten young female rats were selected as young control group. Intragastric administrations were conducted for 4 weeks once daily continuously. The apoptotic rate in rat ovaries were detected by TUNEL. The expression of caspase-3 mRNA and protein in rat ovaries were detected by RT-PCR and Western blot, respectively.</p><p><b>RESULTS</b>The apoptotic rate in rat ovaries in YKN group was lower than that in aged control group, which showed difference between them (P < 0.01). The levels of caspase-3 mRNA and protein in rat ovaries in YKN group were lower than those in aged control group, which showed differences among them (P < 0.01).</p><p><b>CONCLUSION</b>YKN can decrease the apoptotic rate and down-regulate the expression of caspase-3 mRNA and protein in rat ovaries of during perimenopausal period. It may be one of the molecular mechanisms of YKN postponed the ovarian failure and cured perimenopausal syndrome.</p>

Expression of thrombin and its associated protein in cerebellum of human and rat after intracerebral hemorrhage / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Intracerebral hemorrhage (ICH) can cause brain damage through a number of pathways. The purpose of the study was to explore the effect of thrombin, protease nexin-1 (PN-1) and protease activated receptor-1 (PAR-1) in rat and human cerebellum after ICH.</p><p><b>METHODS</b>A model of ICH was produced in adult Sprague-Dawley rats by direct injection of autologous blood (50 microl) into caudate nucleus. Patients with injured hemorrhage were also enrolled in this study. Different expressions of thrombin, PAR-1, PN-1 were detected in rat and human cerebellum by immunohistochemistry and in situ hybridization.</p><p><b>RESULTS</b>In rat cerebellum, thrombin protein significantly increased at 6 hours and reached the maximum 2 days after ICH. The expression of PAR-1 protein reached the maximum at 24 - 48 hours, and then began to decrease. The expression of PN-1 protein reached the maximum at 3 hours, decreased somewhat after that and increased a little at 5 days after ICH. While in human cerebellum, the changing tendency of thrombin, PAR-1 and PN-1 was almost conform to the rat.</p><p><b>CONCLUSION</b>In cerebellum, thrombin can activate PAR-1 expression after ICH, and PN-1 appears quickly after ICH in order to control the deleterious effect of thrombin.</p>

Comparative study on the protective effects of Yinchenhao Decoction against liver injury induced by alpha-naphthylisothiocyanate and carbon tetrachloride / 中国结合医学杂志

<p><b>OBJECTIVE</b>To optimize the animal model of liver injury that can properly represent the pathological characteristics of dampness-heat jaundice syndrome of traditional Chinese medicine.</p><p><b>METHODS</b>The liver injury in the model rat was induced by alpha-naphthylisothiocyanate (ANIT) and carbon tetrachloride (CCl(4) ) respectively, and the effects of Yinchenhao Decoction (, YCHD), a proved effective Chinese medical formula for treating the dampness-heat jaundice syndrome in clinic, on the two liver injury models were evaluated by analyzing the serum level of alanine aminotransferase (ALT), asparate aminotransferase (AST), alkaline phosphatase (ALP), malondialchehyche (MDA), total bilirubin (T-BIL), superoxide dismutase (SOD), glutathione peroxidase (GSH-PX) as well as the ratio of liver weight to body weight. The experimental data were analyzed by principal component analytical method of pattern recognition.</p><p><b>RESULTS</b>The ratio of liver weight to body weight was significantly elevated in the ANIT and CCl(4) groups when compared with that in the normal control (P<0.01). The contents of ALT and T-BIL were significantly higher in the ANIT group than in the normal control (P<0.05,P<0.01), and the levels of AST, ALT and ALP were significantly elevated in CCl(4) group relative to those in the normal control P<0.01). In the YCHD group, the increase in AST, ALT and ALP levels was significantly reduced (P<0.05, P<0.01), but with no significant increase in serum T-BIL. In the CCl(4) intoxicated group, the MDA content was significantly increased and SOD, GSH-PX activities decreased significantly compared with those in the normal control group, respectively (P<0.01). The increase in MDA induced by CCl(4) was significantly reduced by YCHD P<0.05).</p><p><b>CONCLUSION</b>YCHD showed significant effects on preventing liver injury progression induced by CCl(4), and the closest or most suitable animal model for damp-heat jaundice syndrome may be the one induced by CCl(4).</p>

Inhibitory effect of recombinant anti-angiogenic peptide of tumstatin on growth and metastasis of human ovarian cancer transplanted in nude mice / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To evaluate the anti-angiogenic activity of peptide 21 obtained by modification of tumstatin, and its inhibitory effect on the growth and metastasis of human ovarian cancer transplanted in nude mice.</p><p><b>METHODS</b>The peptide 21 was purified by affinity chromatography. Human ovarian cancer SKOV3 cells were inoculated in nude mice and the transplanted tumor was treated with the peptide 21 to observe the tumor growth and metastasis. The microvessel density (MVD) and immunohistochemical staining index of PCNA, VEGF and MMP-2 and TIMP-2 were performed to assess the inhibitory effect of the peptide 21.</p><p><b>RESULTS</b>In the nude mice at 21 days after peptide 21 treatment, the inhibition rate of tumor growth was 53.17%, the tumor microvessel density was significantly reduced (P <0.05), the expression of PCNA, VEGF and MMP-2 were significantly lower (P <0.01), and TIMP-2 expression was significantly higher (P <0.01) in comparison with that of control group.</p><p><b>CONCLUSION</b>The peptide 21 generated in this study has a significant anti-angiogenetic activity, showing significant inhibitory effect on the growth of human ovarian cancer transplanted in nude mice. The mechanism of its inhibitory action on ovarian cancer growth may be mediated by reduction of neovascularization and reduction of expression of angiogenetic factors.</p>

Effects of cloned tumstatin-related and angiogenesis-inhibitory peptides on proliferation and apoptosis of endothelial cells / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Tumstatin is a recently developed endogenous vascular endothelial growth inhibitor that can be applied as an anti-angiogenesis and antineoplastic agent. The study aimed to design and synthesize the small molecular angiogenesis inhibition-related peptide (peptide 21), to replicate the structural and functional features of the active zone of angiogenesis inhibition using tumstatin and to prove that synthesized peptide 21 has a similar activity: specifically inhibiting tumor angiogenesis like tumstatin.</p><p><b>METHODS</b>Peptide 21 was designed and synthesized using biological engineering technology. To determine its biological action, the human umbilical vein endothelial cell line ECV304, the human ovarian cancer cell line SKOV-3 and the mouse embryo-derived NIH3T3 fibroblasts were used in in vitro experiments to determine the effect of peptide 21 on proliferation of the three cell lines using the MTT test and growth curves. Transmission electron microscopy (TEM) and flow cytometry (FCM) were applied to analyze the peptide 21-induced apoptosis of the three cell lines qualitatively and quantitatively. In animal experiments, tumor models in nude mice subcutaneously grafted with SKOV-3 were used to observe the effects of peptide 21 on tumor weight, size and microvessel density (MVD). To initially investigate the role of peptide 21, the effect of peptide 21 on the expression of vascular endothelial growth factors (VEGFs) by tumor tissue was semi-quantitatively analyzed.</p><p><b>RESULTS</b>The in vitro MTT test and growth curves all indicated that cloned peptide 21 could specifically inhibit ECV304 proliferation in a dose-dependent manner (P < 0.01); TEM and FCM showed that peptide 21 could specifically induce ECV304 apoptosis (P < 0.01). Results of in vivo experiments showed that tumors in the peptide 21 group grew more slowly. The weight and size of the tumors after 21 days of treatment were smaller than those in the control group (P < 0.05), with a mean tumor inhibition rate of 67.86%; MVD of the tumor tissue in the peptide 21 group was significantly lower than in the control group (P < 0.05); the number of cells positive for VEGF in the peptide 21 group was significantly fewer than in the control group (P < 0.01).</p><p><b>CONCLUSIONS</b>Similar to the activity of tumstatin in specifically inhibiting tumor angiogenesis, peptide 21 may specifically inhibit tumor endothelial cell proliferation and induce their apoptosis, thereby suppressing tumor angiogenesis and indirectly inhibit the growth, infiltration and metastasis of tumors. Peptide 21 may exert its effect through down-regulating the VEGF expression of tumor cells and vascular endothelial cells.</p>

Role of ERK1/2 kinase in cisplatin-induced apoptosis in human ovarian carcinoma cells / 中国医学科学杂志(英文版)

<p><b>OBJECTIVE</b>To investigate the role of extracellular regulated kinase (ERK1/2) pathway in cisplatin-induced apoptosis in human ovarian carcinoma cells.</p><p><b>METHODS</b>Cisplatin-induced apoptosis were stained with DAPI and was assessed microscopically in human epithelial adenocarcinoma ovarian cell line SKOV3 cells. ERK activation was determined by Western blotting using an anti-phospho-ERK antibody to detect ERK activity. The effect of PD98059 on ERK activity induced by cisplatin was detected by MTT assay.</p><p><b>RESULTS</b>Marked apoptosis of SKOV3 cells resulted from 48 hours treatment with 20 microg/mL cisplatin. Strong activation of ERK was led to by 15 microg/mL cisplatin. Dose response and time course of cisplatin induced apoptosis in SKOV3 cells. Cisplatin-induced ERK activation occurred at 12 hours and increased to highest induction at 24 hours by Western blotting. The effect of PD 98059 on ERK activity induced by cisplatin at the concentration of 100 micromol/L PD 98059. Statistically significant decreased in cell survival were observed with 100 micromol/L PD 98059 at 15 and 20 microg/mL cisplatin (P < 0.05).</p><p><b>CONCLUSIONS</b>Cisplatin activates the ERK signaling pathway in ovarian cancer cell line SKOV3. Inhibition of ERK activity enhances sensitivity to cisplatin cytotoxity in ovarian cancer cell line SKOV3. Evaluation of ERK activity could be useful in predicting which ovarian cancer will response most favorably to cisplatin therapy.</p>

Study on the correlation between the different papillomavirus type and telomerase in cervical cancer / 中华流行病学杂志

<p><b>OBJECTIVE</b>To define a correlation between different human papillomavirus (HPV) types and telomerase activity in cervical cancer.</p><p><b>METHODS</b>Telomerase activity was detected by TRAP-PCR, and different HPV type was determined by PCR in 83 cervical cancer, 47 cervical intraepithelial neoplasia (CIN) and 10 normal cervix cases.</p><p><b>RESULTS</b>With regard to positive rates of telomerase and HPV 16/18: the results were cervical cancer > CIN > normal cervix, CIN III > CIN I, II; with regard to HPV 6/11 positive rate: the results showed CIN I, II > CIN III. Positive rates of telomerase cervical cancer and HPV were bearing on grading and staging, but they did not correlate with histologic subtypes. Positive rate of HPV 6/11 had nothing to do with grading, staging and histologic patterns. On expression strength of telomerase and HPV 16/18: the results showed cervical cancer > CIN, CIN III > CIN I, II. Regard to HPV 6/11'expression strength: the results showed CIN I, II > CIN III, CIN > cervical carcinoma.</p><p><b>CONCLUSION</b>HPV 16/18 infection seemed to have played an important role in carcinogenesis of cervical lesions by activation of telomerase.</p>