Exploring the Workplace Bullying of Indonesian Caregivers and Its Influencing Factors in Taiwan.

Background: Bullying can pose a risk to the health and safety of humans, including the risk of damage to the emotional, psychosocial, mental, or physical health of employees in the workplace. In this study, we aimed to understand the personal characteristics, mental health, sleep quality, and workplace bullying status of Indonesian caregivers and explore the influencing factors of workplace bullying among them. Methods: This cross-sectional study was based on a structured questionnaire in Indonesian, which was designed to collect the data of essential personal characteristics, workplace bullying, sleep quality, and mental health using the Indonesian versions of the Negative Acts Questionnaire−Revised (NAQ-R), Pittsburgh Sleep Quality Index (PSQI), and the Brief Symptoms Rating Scale (BSRS-5). Results: A total of 60.9% of Indonesian caregivers never experienced workplace bullying in Taiwan. A multiple regression analysis revealed that being a household caregiver (ß = 0.14, p = 0.021), sleep quality (ß = 0.18, p = 0.031), and mental health (ß = 0.44, p < 0.001) were significantly correlated with the overall workplace bullying scores of the respondents and revealed that these three variables explained 45% of the variance. Conclusions: Taiwan Indonesian caregivers have a similar workplace bullying rate to Indonesian employees in the workplace. This study indicated the relationships among the workplace bullying of foreign caregivers and demonstrated that being a household caregiver, sleep quality, and mental health were closely related.

Antibacterial, wearable, transparent tannic acid-thioctic acid-phytic acid hydrogel for adhesive bandages.

Making a hydrogel-based first-aid bandage with green resources, desirable biocompatibility, universal adhesive properties, low cost and simple production is a long-standing research aspiration. Considering this, three naturally existing organic acids, namely tannic acid, thioctic acid and phytic acid, were used to construct a novel adhesive gel (TATAPA hydrogel) for epidermal tissue bandage applications. This hydrogel could be synthesized under mild conditions with no need for a freeze-thawing shaping procedure, and was transparent, moldable and stretchable with good stability under continuous water immersion. In lap-shear tests, the TATAPA hydrogel could adhere to various hydrophilic and hydrophobic surfaces. Moreover, in the case of skin tissue adhesion, the hydrogel could be easily peeled off from the skin, meeting wearability requirements. Rheological tests showed that the hydrogel possessed thermal sensitive properties derived from multi-supramolecular interactions. The methicillin-resistant Staphylococcus aureus (MRSA)-infected burn wound test demonstrated that the hydrogel had desirable antibacterial activity and was beneficial for wound healing. A femoral artery bleeding assay was also used to reveal that the TATAPA hydrogel could be directly pasted onto the bleeding site for hemostasis. Overall, this hydrogel demonstrates potential as a surgical bioadhesive for a broad range of medical applications.

[Antitumor and antimetastatic effect of antimicrobial peptide conjugated with tumor homing peptide TMTP1 on the transplanted prostate cancer and gastric cancer in nude mice].

OBJECTIVE: Due to their lower risk for induction of resistance, antimicrobial peptides with selective anticancer effect could be developed into a new generation of anticancer drugs. We conjugated an antimicrobial peptide with tumor-targeting peptides (TMTP1) to explore whether it has inhibiting effect on the progression and metastasis of transplanted prostate cancer and gastric cancer in nude mice. METHODS: Subcutaneously transplanted human prostate cancer and orthotopically transplanted human gastric cancer in nude mice were prepared. 50 µmol/L PBS (control group), 50 µmol/L TMTP1 (TMTP1 group) or 50 µmol/L TMTP1-GG-D(KLAKLAK)(2) (treatment group) were injected i.p. to the three groups of nude mice, respectively. The binding ability of the novel fusion polypeptide TMTP1-GG-D(KLAKLAK)(2) to the tumors and its antitumor effect were assessed by measurement of tumor volume, histopathological examination of the tumor tissues, testing apoptosis index of tumor cells with TUNEL staining, and survival curve plotting of the mice. RESULTS: The median survival time of subcutaneous prostate cancer-bearing mice was 50 days in the control group, 55 days in the TMTP1 group, and 70 days in the TMTP1-GG-D(KLAKLAK)(2) group (P < 0.05). The median survival time of the subcutaneous gastric cancer-bearing mice was 25 days in the control group, 30 days in the TMTP1 group, and 45 days in the TMTP1-GG-D(KLAKLAK)(2) group (P < 0.01). The tumor volume in the subcutaneous prostate cancer-bearing mice was (2.5 ± 0.3)cm(3) in the control group, (1.8 ± 0.2) cm(3) in the TMTP1 group, and (0.3 ± 0.1)cm(3) in the TMTP1-GG-D(KLAKLAK)(2) group (P < 0.01). The tumor volume of the subcutaneous gastric cancer-bearing mice was (3.8 ± 0.4) cm(3) in the control group, (3.2 ± 0.2)cm(3) in the TMTP1 group, and (0.4 ± 0.1) cm(3) in the TMTP1-GG-D(KLAKLAK)(2) group (P < 0.01). Large tumors were observed in the stomach of the orthotopic gastric cancer-bearing mice of the control and TMTP1 groups. The tumor volume of the TMTP1-GG-D(KLAKLAK)(2) group was obviously reduced. White metastases in the liver, spleen and abdominal wall were observed in the control and TMTP1 groups (P < 0.01). TUNEL staining revealed that the apoptosis index of the control group was (31.9 ± 1.5)%, TMTP1 group (37.2 ± 2.3)% and TMTP1-GG-D(KLAKLAK)(2) group (69.7 ± 2.1)% (P < 0.01). CONCLUSIONS: The results of our study demonstrate that the novel fusion peptide of antimicriobial peptide conjugated with TMTP1 can effectively inhibit tumor progression and metastasis, therefore, is promising to be a novel effective anticancer drug.

Case-control study of diet in patients with cervical cancer or precancerosis in Wufeng, a high incidence region in China.

PURPOSE: To investigate the diet of patients with cervical cancer and precancerosis in the Wufeng area, a high- incidence region in China. METHODS: In the case group, 104 patients diagnosed with cervical cancer or cervical intraepithelial neoplasias (CINII/III) were recruited from the Wufeng area. Nine hundred thirty-six healthy women were selected from the same area as the matched controls. A questionnaire, which included questions about general lifestyle conditions, smoking and alcohol status, source of drinking water, green tea intake, and diet in the past year, was presented to all participants. RESULTS: Green tea intake (P=0.022, OR=0.551, 95% CI=0.330-0.919) and vegetable intake (P=0.035, OR=0.896, 95% CI=0.809-0.993) were identified as protective factors against cervical cancer or CINII/III. There was no indication of any associations of other lifestyle factors (smoking status, alcohol status, source of drinking water) or diet (intake of fruit, meat/egg/milk, soybean food, onion/garlic, staple food and pickled food) with cervical cancer. CONCLUSIONS: The results suggest that eating more fresh vegetables and drinking more green tea may help to reduce the risk of cervical cancer or CINII/III in people of the Wufeng area.

[Preparation of HPV18 E7 peptide plus CpG vaccine and its immunologic effects in vitro].

OBJECTIVE: To screen and prepare the vaccine of human papillomavirus (HPV) 18 E7 peptide target at human leukocyte antigen (HLA)-A2 plus CpG through SYFPEITHI. METHODS: (1) The SYFPEITHI database was employed for predicting and screening of HPV18 E7 HLA-A2 restricted T cell epitopes.(2) The peripheral blood and tumor tissue sample of HLA-A2 positive and HPV18 positive/negative patients were collected and randomly divided into 7 groups, i.e. E7PA + CpG, E7PB + CpG, E7PC + CpG, E7PD + CpG, CpG, IR-T + CpG and control groups respectively. T cell proliferation was detected by thiazolyl blue tetrazolium bromide (MTT) assay at different timepoints. Lactate dehydrogenase delivery method (LDH) was used to test the cytolytic t lymphocyte (CTL) activity of peripheral blood mononuclear cell (PBMC) in different ratios of effect and target (E:T). And the level of activity T cells was evaluated by interferon gamma (IFN-γ)-related enzyme-linked immuno-spot assay (ELISPOT). RESULTS: (1) Four peptides named E7PA, E7PB, E7PC and E7PD were obtained separately with high levels of affinity and specificity. (2) During continuous observations after vaccination, the E7PA + CpG group had the most pronounced proliferation rate. When E:T = 100:1, the E7PA + CpG group had more powerful CTL effect than the control group with statistic significance (P < 0.00). E:T was concentration-dependent. Except for IR-T + CpG, all other groups had great difference than control group with statistic significance (P < 0.05) but no significant difference between the groups. The levels of IFN-γ spot-forming T cells were higher in the E7PA + CpG group than the control group with statistic significance (P < 0.01). In terms of specificity, E7PA + CpG in the HPV18 positive group could induce the proliferation of IFN-γ-secreting T cells. And there was statistical difference with the control group (P < 0.05). CONCLUSION: Screening the HPV18 E7 peptide target at HLA-A2 plus CpG as the candidate targets by SYFPEITHI may active specific immunological cellular responses to HPV18 positive disease.

Diagnostic dilemma of abdominopelvic tuberculosis:a series of 20 cases.

OBJECTIVE: To establish more effective diagnostic procedures to identify the characteristic features of abdominopelvic tuberculosis (APTB) mimicking advanced ovarian cancer. METHODS: A retrospective review of 20 cases of APTB mimicking advanced ovarian cancer was undertaken. RESULTS: The mean age of the patients was 28.9 ± 10.8 years. The main clinical manifestations were abdominal pain (45%) and distention (45%). CA125 level was elevated in 18 cases (90.0%). Pelvic mass in 18 patients (90.0%) and ascites in 12 patients (60.0%) were detected by using abdominal US. The bacteriologic cultures and cytological studies were all negative (10 cases, 100%). Laparotomy (17 cases) and laparoscopic evaluation (1 case) was performed with the presumptive diagnosis of advanced ovarian cancer except for 2 patients treated with diagnostic anti-TB chemotherapy. The common intra-operative findings were miliary nodules (14 cases, 77.8%) and widespread adhesion (10 cases, 55.6%). Intra-operative frozen section was obtained in 10 cases, and the typical tuberculosis tubercles were detected in all cases. CONCLUSION: APTB should be considered in all cases with pelvic mass, ascites and high levels of CA125, although clinical features and laboratory results specifically indicate neither ovarian malignancy nor APTB. Diagnostic laparotomy is a direct and safe method. To avoid extended surgery, the cases with APTB can be diagnosed through intra-operative frozen section in conjunction with clinical features.

[Effect of HPV16 peptide vaccine in combination with paclitaxel-cisplatin chemotherapy on cervical cancer in vitro and in vivo].

OBJECTIVE: to the explore the effect of Human papillomavirus (HPV) 16 peptide vaccine in combination with paclitaxel-cisplatin (TP) chemotherapy on cervical cancer in vitro and in vivo. METHODS: (1) the major histocompatibility complex (MHC) class I restricted T cell epitopes were studied by bioinformatics for transporter associated with antigen processing (TAP). Their effects were compared and E7Pa had the most dramatic effect. (2) In vivo, the C57BL/6 mice were divided equally into 6 groups randomly after loading with TC-1 cells (HPV 16 positive tumor cells from C57BL/6 mouse), named as E7Pa + CpG + TP, E7Pa + CpG, CpG + TP, TP and CpG group as experiment groups and control (blank injection with physiological saline). The tumor volumes were measured regularly by tumor growth curve to compare the therapeutic effects in different groups; the related cell factors in murine peripheral blood were evaluated by enzyme-linked immunosorbent assay (ELISA); the TUNEL test kit was used to explore cellular apoptosis in tumor tissue; the survival curve was drawn from the TC-1 cell loading to natural death; safety was tested by pathological test and leucocyte count. RESULTS: at day 60 of tumor growth, the tumor volume of immunotherapy plus TP chemotherapy group was (0.013 ± 0.010) cm(3) versus the control (1.900 ± 0.075) cm(3) with a great significant deviation (P < 0.01). Meanwhile, the volumes were E7Pa + CpG group (0.340 ± 0.038) cm(3), TP + CpG group (0.650 ± 0.029) cm(3), TP group (1.100 ± 0.052) cm(3) and that of CpG group was (0.890 ± 0.047) cm(3) separately. And these groups had significant difference with the controls (P < 0.05). The average survival time of different groups were E7Pa + CpG + TP group (108.50 ± 8.97) d, E7Pa + CpG group (100.02 ± 2.27) d, CpG + TP group (79.63 ± 4.05) d, TP group (73.24 ± 3.11) d, CpG group (68.63 ± 1.38) d and controls (52.37 ± 2.47) d. And the difference between the E7Pa + CpG + TP and E7Pa + CpG groups had great significance with the controls (P < 0.01). Furthermore, the immune system was effectively stimulated for suppressing tumor growth in the immunotherapy group while cell apoptosis was significantly induced in the chemotherapy group. The combination of immunotherapy and chemotherapy was significantly efficacious than either of them alone. And it could thoroughly stimulate the immune effects and enhance the anti-tumor function of chemotherapeutical drugs. In safety test, there was no significant difference among all groups. CONCLUSION: the HPV16 peptide vaccine in combination with TP chemotherapy can treat the HPV16 E7 positive tumor effectively in experiment.

[Clinical analysis of 132 cases of cervical adenosquamous carcinoma and cervical adenocarcinoma].

BACKGROUND AND OBJECTIVE: The incidence of cervical adenosquamous carcinoma is relatively low. This study was to analyze the clinicopathologic characteristics and prognostic factors of cervical adenosquamous carcinoma. METHODS: Clinical data of 44 cervical adenosquamous carcinoma patients and 88 cervical adenocarcinoma patients(control), treated from January 2002 to December 2007, were analyzed using Chi-square test, Kaplan-Meier method, log-rank test, and Cox regression model. RESULTS: The proportion of large tumors (maximal diameter > 4 cm) was significantly higher in cervical adenosquamous carcinoma group than in cervical adenocarcinoma group (47.7% vs. 28.4%, P<0.05); the proportion of poorly differentiated tumors was significantly higher in cervical adenosquamous carcinoma group than in cervical adenocarcinoma group (56.8% vs. 30.7%, P<0.05). Univariate analysis showed that tumor size (P=0.011), FIGO stage (P=0.013), depth of stromal invasion (P=0.05) and lymph node metastasis (P=0.017) were correlated with prognosis, while multivariate analysis showed that FIGO stage and lymph node metastasis had great impact on prognosis. There was no significant difference of 2-year overall and disease-free survival rates between the two groups (P>0.05). CONCLUSIONS: Cervical adenosquamous carcinoma is characterized by large tumor size and poor differentiation. FIGO stage and lymph node metastasis are significant prognostic factors. There is no difference in prognosis between cervical adenosquamous carcinoma and cervical adenocarcinoma.

[Down-regulation of Chk1/Chk2 gene expression increases apoptosis in irradiated HeLa cells and its mechanism].

OBJECTIVE: To explore the increasing effect of blocking Chk1 and /or Chk2 gene by Chk1 or Chk2-specific antisense oligodeoxynucleotides (AsODN) on apoptosis in HeLa cell line after irradiation and its mechanism of action. METHODS: Asynchronized HeLa cells were exposed to (60)Co-irradiation at different dosage to activate G(2)/M checkpoint arrest. The cell cycle profiles were observed in HeLa cells after irradiation at a range of various doses and different time points by flow cytometry. In the experimental groups, Chk1/2 sODN and AsODN alone or in combination were transfected into HeLa cells, and the cells were exposed to (60)Co-irradiation at 24 h after transfection. The changes of Chk1/2 protein expression were assayed by Western blot and confocal laser scanning microscopy (Confocal), and the cell cycles, apoptosis rates and cell cycle specific apoptosis were detected by annexin V-PI labeling and flow cytometry. RESULTS: Apoptotic response was significantly increased in the Hela cells after G(2)/M arrest and was inversed to activation of G(2)/M checkpoint. Either Chk1 or Chk2-specific AsODN consistently enhanced DNA damage-induced apoptosis by 90% approximately 120%, compared to corresponding sODN control (P < 0.05). Unexpectedly, combined use of Chk1- and Chk2-specific AsODN did not produce synergistic effect as compared to treatment with Chk1- or Chk2-specific AsODN alone (P > 0.05). While irradiated HeLa cells underwent apoptosis preferentially in G(1)-phase, apoptosis occurred in either of G(1)-, S- or G(2)/M -phase in the presence of Chk1 and/or Chk2 AsODN. CONCLUSION: The radioresistance is mainly induced by activating the cell cycle checkpoint signal transduction pathway after irradiation, and abrogating of the key effector Chk1 and Chk2 may increase the apoptotic sensitivity to irradiation due to changes of the pattern of cell cycle specific apoptosis.

[Construction of anti-sense cDNA library of human breast cancer cells during apoptosis induced by trichostatin A and preliminary screening of essential genes].

OBJECTIVE: To construct an anti-sense cDNA library of human breast cancer cells to screen essential genes with anti-tumor effects on apoptosis of human breast cancer cells induced by trichostatin A. METHODS: Poly (A)(+)RNA was extracted from human breast cancer cells of the line MCF-7 treated by trichostatin A for 0, 12, 24, 36, 48, 60, or 72 h. cDNA were synthesized and inserted reversely into PCEP 4 vector to construct an anti-sense cDNA library. HeLa cells were transfected with the library DNA or blank PCEP 4 vector as control group. All the transfected cells were screened by 200 nmol/L trichostatin A and 200 microg/ml hygromycin B. Screening was stopped when the control cells died. Then the surviving cell clones were amplified and Hirt DNA was extracted. Several expressed sequence tags were thus obtained. The data were analyzed by bioinformatics and interested EST fragment was chosen for preliminary functional screening. RESULTS: An anti-sense cDNA library was constructed containing 2 x 10(6) independent clones with an insert efficiency of more than 90%; DNA sequencing and bioinformatic analysis suggested that the No.27 survival clone was zinc transporter LIV1 showing a strong resistance against trichostatin A-induced apoptosis during functional screening. CONCLUSION: An anti-sense cDNA library with high quantity and quality has been successfully constructed; LIV1 gene may be one of the essential genes with anti-tumor effects on apoptosis induced by trichostatin A.

[Inhibitory effects of an antisense PCDGF vector on proliferation and invasion of highly malignant ovarian cancer cells and the related mechanism].

OBJECTIVE: To investigate the inhibitory effects of an antisense PC cell derived growth factor (PCDGF) vector on proliferation and invasion of highly malignant ovarian cancer cell lines Sw626 and A2780 cells, and preliminarily explore the related mechanisms. METHODS: MTT assay and Boyden chamber in vitro invasion assay were employed to detect the changes of proliferation and invasion ability in the Sw626 and A2780 cells transfected with anti-sense PCDGF. The expression levels of cyclin D1 and CDK4 proteins before and after transfection were detected by Western blotting. The effects on the expression and activity of MMP-2 were evaluated by quantitative RT-PCR and zymography, respectively. RESULTS: Comparing with the blank group, the proliferation inhibition rate of the Sw626 and A2780 cells transfected with anti-sense PCDGF was 72.9% and 70.9%, respectively, and the invasion ability was inhibited by 62.9% and 59.0%, respectively. The levels of cyclin D1 and CDK4 protein expression in antisense PCDGF transfected cells were 0.38 +/- 0.08 and 0.37 +/- 0.13, respectively, all significantly lower than 0.84 +/- 0.11 and 0.64 +/- 0.11, respectively, in the blank group (P < 0.01). The MMP-2 mRNA expression level in antisense PCDGF transfected cell group was 0.66 +/- 0.11, not significantly decreased in comparison with 0.89 +/- 0.09 in the blank group (P > 0.05), but the activity of MMP-2 was inhibited significantly. CONCLUSION: The antisense PCDGF vector may inhibit markedly the proliferation and invasion of highly malignant ovarian cancer cells, and partially reverses their malignant phenotype. It seems to be related with down-regulating the expression of cyclin D1 and CDK4 and inhibiting the activity of MMP-2. Our findings indicate that PCDGF may become a new target for antisense gene therapy of ovarian cancer.

[Silencing of cell cycle checkpoint kinase gene enhances cisplatin-induced apoptosis of lung cancer cells].

OBJECTIVE: To investigate the changes in cell cycle induced by cisplatin (DDP) and the effect of antisense oligonucleotide (AsODN) targeting Chk1/2 on DDP-induced apoptosis in lung cancer cell line A549 cells. METHODS: The characteristics of cell cycle and apoptosis induced by DDP were detected by flow cytometry using SubG1 method. Chk1/2 mRNA and protein expression were assayed by RT-PCR and Western blot under best condition of transfection of AsODN targeting Chk1/2 by lipofection. Apoptosis of A549 cells induced by DDP was determined by flow cytometry using AnnexinV-FITC staining after transfection of Chk1/2 AsODN. RESULTS: Asynchronized A549 cells were treated with 10 micromol/L DDP, and significant S-phase arrest was observed at 12 h later. Transfection with antisense oligonucleotide targeting Chk1/2 inhibited the Chk1/2 expression at both mRNA and protein levels. Either Chk1 or Chk2-specific AsODN consistently enhanced DNA damage-induced apoptosis by 100% - 200%, compared with that in the sODN control (P < 0.05), but combined use of Chk1- and Chk2-specific AsODN did not show synergistic effects as compared with that induced by treatment with Chk1- or Chk2-specific AsODN alone (P > 0.05). CONCLUSION: Chk1 and Chk2 may be regarded as effective targets of chemotherapy for lung cancer. Silencing the key effector Chk1 and Chk2 genes may significantly increase the chemosensitivity of lung cancer cells.

[Preparation of human papillomavirus 16 E7 peptide vaccine and its effectiveness in vitro and in vivo.].

OBJECTIVE: To prepare the human papillomavirus (HPV) 16 peptide vaccine and explore the effect in vitro and in vivo. METHODS: (1) Prediction of the major histocompatibility complex (MHC) class I restricted T cell epitopes by bioinformatics target at transporter associated with antigen processing (TAP) and named by E7Pa, E7Pb, E7Pc separately. (2) In vivo, the C57BL/6 mice were divided into five groups with same amounts randomly after loading with TC-1 cells (HPV 16 positive tumor cells from C57BL/6 mouse), named as E7Pa + CpG, E7Pb + CpG, E7Pc + CpG (as experiment groups, and added 50 microg/ml E7Pa, E7Pb, E7Pc, respectively), CpG (as positive control group and added Con A with 12 mg/L final concentration) and blank control group (without any treatment). The T cell proliferation was detected by methyl thiazolyl tetrazolium (MTT) assay at different time points; the lactate dehydrogenase (LDH) delivery method was used to test the cytolytic T lymphocyte (CTL) activity of mouse splenic lymphocyte in different ratio of effector cells and target cells (E:T); the related cytokines in tumor tissue and mouse peripheral blood were evaluated by real-time PCR and enzyme-linked immunosorbent assay (ELISA), respectively. The tumor volumes were measured to contrast the therapeutic effect in different groups. RESULTS: (1) Three peptide named E7Pa, E7Pb, E7Pc were successfully preparated which had high affinity and specificity. (2) After vaccination of 24, 48, 72, 96 hours, MTT results shown that the proliferation rate in E7Pa + CpG group were (131 +/- 32)%, (302 +/- 15)%, (552 +/- 28)%, (731 +/- 24)% individually, which were much higher than those in blank control [(72 +/- 15)%, (120 +/- 57)%, (176 +/- 41)%, (288 +/- 29)%; P < 0.01], and the other groups i.e. E7Pb + CpG, E7Pc + CpG and CpG groups all proliferated much higher than those in blank control group with statistic signification (P < 0.05), but there was no significant difference between groups (P > 0.05); the LDH delivery assay showed that when the ratio of E:T was 100:1, the activity of CTL in the E7Pa + CpG group was most powerful than the other groups with statistic signification (P < 0.01).Meanwhile, the ratio of E:T was concentration-dependent. Compared E7Pb + CpG, E7Pc + CpG or CpG groups with blank control group, there were significantly difference (P < 0.05), while there was no significant difference between groups (P > 0.05). The mRNA levels of interferon gamma (IFN-gamma), interleukin-2 (IL-2) in tumor tissue and peripheral blood in E7Pa + CpG group were significantly higher than those in blank control group (P < 0.01), which was the similar results when compared E7Pb + CpG, E7Pc + CpG or CpG groups with control group (P < 0.05), and without significant difference between groups (P > 0.05). The tumor volumes were suppressed obviously in all the experiment groups, especially at the 60th days, the volumes in E7Pa + CpG group were much smaller than that in blank control group with statistic signification (P < 0.01), which was the similar results that E7Pb + CpG, E7Pc + CpG or CpG groups had difference than blank control group with statistic signification (P < 0.05), and without significant difference between groups (P > 0.05). CONCLUSION: The HPV16 E7 peptide target at TAP combination with CpG as a vaccine could treat effectively the HPV16 E7 positive tumor in experiment.

Antisense Ki-67 cDNA transfection reverses the tumorigenicity and induces apoptosis in human breast cancer cells.

Effects of antisense Ki-67 cDNA transfection on breast cancer cells were investigated in this study. Transfection of antisense Ki-67 cDNA resulted in a 70%-80% reduction in proliferation of MDA-MB-435s, cells which highly expressed Ki-67 mRNA and protein. Transwell assay showed that mobility and invasion capability was dramatically inhibited by 50%-60%, and cell cycle analysis displayed a higher proportion in G(2)/M and G(0)/G(1) phases accompanied by remarkably increased ratio of apoptotic cells. Our results suggested that antisense Ki-67 cDNA vector treatment might be an important potential option in the anticancer therapy.

[Correlation of aquaporin 1 with hypoxia-inducible factor 1 in breast cancer].

OBJECTIVE: To determinate the correlation of aquaporin 1 (AQP1) and hypoxia-inducible 1 (HIF-1). METHODS: 155 samples of breast cancer obtained during radical mastectomy underwent immunohistochemistry to detect the expression of AQP1 and HIF-1. RESULTS: The positive rates of AQP1 in the HIF1 positive group was 297 +/- 25, significantly higher than that of HIF1 negative group (168 +/- 38, P < 0.001). CONCLUSION: AQP1 is positively correlated with HIF1. The interaction of AQP1 and HIF1 may co-regulate the progress of breast cancer.

[Effects of vimentin on invasion and metastasis of prostate cancer cell lines PC-3M-1E8 and PC-3M-2B4].

BACKGROUND & OBJECTIVE: Vimentin, a cytoskeleton protein, is involved in regulating the migration and proliferation of cells. This study was to detect the expression of vimentin in prostate cancer cell lines PC-3M-1E8 and PC-3M-2B4, and evaluate the effects of vimentin on invasion and metastasis of prostate cancer cells. METHODS: Two-dimensional gel electrophoresis (2-DE) followed by matrix-assisted laser desorption/time of flight mass spectrometry (MALDI TOF-MS) were used to detect the expression of vimentin in prostate cancer cell lines PC-3M-1E8 and PC-3M-2B4. Genetic intervention of vimentin gene and in vitro invasion assay were used to investigate the effects of vimentin on the invasion and metastasis of prostate cancer cells. RESULTS: The protein level of vimentin was higher in PC-3M-1E8 cells with high metastatic potential than in PC-3M-2B4 cells with low metastatic potential. Eukaryotic vectors expressing antisense-and sense-vimentin were constructed successfully and transfected into PC-3M-1E8 and PC-3M-2B4 cells separately. The number of invasive cells was significantly lower in PC-3M-1E8/vas cells with antisense-vimentin than in control PC-3M-1E8/3.1(-) cells (99.3+/-4.8 vs. 319.4+/-6.5, P<0.01), and significantly higher in PC-3M-2B4/vs cells with sense-vimentin than in control PC-3M-2B4/3.1(+) cells (330.5+/-5.8 vs. 98.6+/-7.5, P<0.01). CONCLUSION: Vimentin is a promising marker for predicting the invasion and metastasis of prostate cancer cells.

[Reversal of drug resistance in human ovarian cancer cells by wild-type PTEN gene and its mechanisms].

OBJECTIVE: To examine expression of PTEN gene in ovarian cancer cisplatin-sensitive cell line OV2008 cells and cisplatin-resistant cell line C13K cells, and evaluate the effect of wild-type PTEN gene on reversing cisplatin-resistance of C13K cells and underlying mechanisms. METHODS: The expression of PTEN mRNA and protein in OV2008 and C13K cells were detected by semi-quantitative RT-PCR and western blot. Recombinant eukaryotic expression plasmid containing human wild-type PTEN gene was transfected into C13K cells by lipofectamine 2000. The expression of PTEN mRNA was monitored by RT-PCR and the expression of PTEN, protein kinase B (AKT), phospho-AKT (p-AKT) protein were analyzed by western blot in PTEN transfected and untransfected C13K cells. Proliferation and chemosensitivity of cells to cisplatin were measured by methyl thiazolyl tetrazolium (MTT), and cell apoptosis was detected by flow cytometry after treatment with cisplatin. RESULTS: (1) The expression of PTEN mRNA and protein (1.02 +/- 0.05, 1.02 +/- 0.07) in OV2008 cells were significantly higher than those in C13K cells, which were 0.45 +/- 0.03 and 0.55 +/- 0.03 respectively (P < 0.05). (2) After transfected with PTEN gene for 48 hours, the expression of PTEN mRNA and protein in C13K cells were 2.04 +/- 0.10, 0.94 +/- 0.04 respectively. Compared with C13K cells transfected with empty vector (1.04 +/- 0.04, 0.36 +/- 0.03) and untransfected C13K cells (1.03 +/- 0.05, 0.37 +/- 0.03), the difference was significant respectively (P < 0.01). The expression of p-AKT protein (0.94 +/- 0.07) was lower than those in control groups (1.66 +/- 0.10, 1.68 +/- 0.14; P < 0.05). (3) The 50% inhibition concentration (IC(50)) to cisplatin of C13K cells transfected with PTEN [(7.2 +/- 0.3) micromol/L] was obviously lower than those of empty-vector transfected cells and untransfected cells [(12.7 +/- 0.4), (13.0 +/- 0.3) micromol/L; P < 0.05]. (4) The apoptosis ratio of C13K cells with wild-type PTEN transfection, empty vector transfection and untransfected were (41.7 +/- 0.9)%, (18.6 +/- 0.7)% and (15.3 +/- 0.8)% respectively (P < 0.01). CONCLUSIONS: PTEN gene plays an important role in ovarian cancer multidrug resistance. Transfection of PTEN could increase the expression of PTEN and restore drug sensitivity to cisplatin in multidrug-resistant human ovarian cancer cell line C13K by decreasing the expression of p-AKT.

[Role of human telomerase reverse transcriptase in apoptosis of human umbilical vein endothelial cells induced by trichostatin A].

OBJECTIVE: The aim of this study was designed to investigate the effect of TSA on human umbilical vein endothelial cells and to reveal its possible mechanisms and relationship between apoptosis and activity of telomerase reverse transcriptase. METHODS: sulforhodamine B method was employed to determine the growth rate of umbilical vein endothelial cells. The cell apoptotic rate was measured by flow cytometry (FCM). The hTERT and p21(Waf1) mRNA expression before and after TSA treatment were detected by semiquantitative reverse transcription polymerase chain reaction (RT-PCR). The quantitative of hTERT protein expression in cells were detected by flow cytometry. After transfection, the cell telomerase activity was detected by PCR and telomeric repeat amplification protocol assay (PCR-TRAP-ELISA) and early apoptosis was measured by Annexin V/PI stain and flow cytometry. RESULTS: After being treated with TSA, the proliferation of umbilical vein endothelial cells was inhibited. Slight apoptosis and cell cycle arrest were detected. However, the same concentration of TSA induced serious apoptosis in HeLa cells. Up-regulation of hTERT mRNA expression was observed within 48 h after TSA treatment, but the change of p21(Waf1) expression was not significant. The umbilical vein endothelial cells hTERT protein expression level was increased within 24 h. After transfection of the dominant negative, wild type and control hTERT plasmid, a significant difference of telomerase activity in these cells was observed by PCR-TRAP-ELISA assay. WT-hTERT-transfected cells were more resistant to apoptosis induced by trichostatin A. CONCLUSION: Human umbilical vein endothelial cells could be resistant to apoptosis induced by high concentrate TSA, and hTERT might play an important role in this process.

[Antisense targeting to human papillomavirus 18 E6/E7 affects the proliferation and apoptosis of human cervical carcinoma: an in vitro experiment with HeLa cells].

OBJECTIVE: To investigate the effect of eukaryotic fluorescent expression vector carrying antisense human papillomavirus (HPV) 18 E6/E7 on the growth and proliferation of human cervical carcinoma. METHODS: The HPV18 E6E7 with the length of 716 bp was amplified by PCR, the PCR product was inversely inserted into the eukaryotic fluorescent expression vector pEGFP-C1 so as to construct the recombinant eukaryotic expression plasmid pEGFP-HPV18E6E7as (EGFP-18AS). Human cervical carcinoma cells of the line HeLa were cultured and randomly divided into 3 groups: Group A transfected with the recombinant plasmid EGFP-18AS, Group B transfected with the blank plasmid pEGFP-C1, and Group C without transfection used as control group. The mRNA expression of HPV 18 E6/E7 in the HeLa cells was detected by RT-PCR and protein expression of HPV18 E6/E7 HPV 18 E6/E7 in the HeLa cells was detected by. Western blotting MTT assay was performed to dynamically monitor the surviving cells and the cell apoptosis was observed by flow cytometry and fluorescence microscopy. RESULTS: The protein and mRNA expression levels of HPV18 E6/E7 in the HeLa cells transfected with HeLa/18AS were both remarkably lower than those in the HeLa cells transfected with blank plasmid and those of the control group. The numbers of surviving HeLa cells of Group A was significantly lower than those of Groups B and C (both P < 0.05). The phenomenon of arrest of G(1) phase was remarkable in Group A. The apoptotic rate of the cells of Group A was 47.21%, significantly higher than those of Groups B and C (14.18% and 3.36% respectively, both P < 0.05). An increased number of cells with chromosome condensation and fragmentation was found in Group A as compared with Groups B and C. CONCLUSION: The recombinant pEGFP-HPV18E6E7as can effectively inhibit the growth and proliferation of human cervical carcinoma HeLa cells, and further induce the cell apoptosis. The antisense RNA technology is available and may provide a new way to gene therapy of the cervical carcinoma.

[Effect of hypoxia on the chemotherapeutic sensitivity of human ovarian cancer cells to paclitaxel and its mechanism].

OBJECTIVE: To explore the effect of hypoxia and hypoxia-inducible factor-1alpha (HIF-1alpha) on the expression of multidrug resistance gene-1 (mdr-1) and its coded p-glyeoprotein (P-gp) as well as the chemotherapeutic sensitivity of human ovarian cancer cells to paclitaxel and its mechanism. METHODS: The mRNA and protein levels of HIF-1alpha, mdr-1 and p-gp were studied by immunocytochemistry, semiquantitative reverse transcription-ploymerase chain reaction (RT-PCR) and Western blot analysis in human ovarian cancer cells (A2780) in 5% CO2 + 1% O2 hypoxic culture and 21% O2 normoxic culture, respectively. Methyl thiazolyl tetrazolium (MIT) was used to evaluate the chemotherapeutic sensitivity of A2780 cells to paclitaxel by inhibition rate. RNA interference technique was used and small hairpin RNAs (shRNAs) eukaryotic expression vector targeting HIF-1alpha was constructed as pSiHIF-1alpha, and transfected into A2780 cells. RT-PCR and Western blot were used to detect gene silencing effect on HIF-1alpha, the expressions of mdr-1 and p-gp. The inhibition rate was observed after HIF-1alpha gene silence. RESULTS: HIF-1alpha at both mRNA and protein levels was induced significantly under hypoxia. The HIF-1alpha expression at mRNA level was oxygen gradient-independent, while HIF-1alpha expression at protein level was oxygen gradient-dependent. The inhibition rate of paclitaxel to hypoxic A2780 cells in 5% CO2 + 1% O2 was significantly lower than that in normoxic A2780 cells (P <0.05). The shRNAs plasmid targeting HIF-1alpha was constructed successfully and HIF-1alpha gene was silenced in A2780 cells efficiently followed by mdr-1 and p-gp down-regulation. The inhibition rate was greatly increased in hypoxic A2780/siHIF-1alpha cells. CONCLUSION: Hypoxia can decrease the chemotherapeutic sensitivity of human ovarian cancer A2780 cells to paclitaxel through HIF-1alpha regulating the expression of mdr-1 and p-gp.