Clinical analysis of 13 males with primary choriocarcinoma and review of the literature.

OBJECTIVE: To analyze the management and prognosis of primary choriocarcinoma (PCC) in male patients. METHODS: The clinical records of males with PCC who were treated at Peking Union Medical College Hospital between 1990 and 2012 were analyzed retrospectively. The literature regarding this clinical condition was also reviewed. RESULTS: The median survival interval of the 13 patients treated at Peking Union Medical College Hospital was 54 months (range, 6-115 months), and the 1- and 3-year survival rates were 53.8% and 43.1%, respectively. All patients were treated with surgery; 12 were treated with combined chemotherapy. After including 100 cases found in the literature, for a total of 113 patients, the median survival interval was 10 months (range, 6.4-13.6 months). The testis was the most common primary site (36.2%). Most patients (70.9%) had metastatic lesions at diagnosis. Univariate and multivariate analyses revealed that longer median overall survival was significantly associated with patient age <34 years old (48 months vs 10 months, odds ratio [OR] =0.47, P=0.029), the presence of other histological components (54 months vs 11 months, OR =0.54, P=0.011), and combined chemotherapy and surgical treatments (14 months vs 2.5 months, OR =0.18, P=0.002). CONCLUSION: PCC is an extremely rare disease among men, and its prognosis is much worse than that of gestational choriocarcinoma. The complete resection of the primary site and metastases followed by chemotherapy seems to provide patients with the best chance at survival. Furthermore, additional chemotherapy cycles might facilitate better progress.

Expression of the CXCL12/CXCR4 and CXCL16/CXCR6 axes in cervical intraepithelial neoplasia and cervical cancer.

The chemokine CXCL12 is highly expressed in gynecologic tumors and is widely known to play a biologically relevant role in tumor growth and spread. Recent evidence suggests that CXCL16, a novel chemokine, is overexpressed in inflammation-associated tumors and mediates pro-tumorigenic effects of inflammation in prostate cancer. We therefore analyzed the expression of CXCL12 and CXCL16 and their respective receptors CXCR4 and CXCR6 in cervical intraepithelial neoplasia (CIN) and cervical cancer and further assessed their association with clinicopathologic features and outcomes. Tissue chip technology and immunohistochemistry were used to analyze the expression of CXCL12, CXCR4, CXCL16, and CXCR6 in healthy cervical tissue (21 cases), CIN (65 cases), and cervical carcinoma (60 cases). The association of protein expression with clinicopathologic features and overall survival was analyzed. These four proteins were clearly detected in membrane and cytoplasm of neoplastic epithelial cells, and their distribution and intensity of expression increased as neoplastic lesions progressed through CIN1, CIN2, and CIN3 to invasive cancer. Furthermore, the expression of CXCR4 was associated significantly with the histologic grade of cervical carcinoma, whereas the expression of CXCR6 was associated significantly with lymph node metastasis. In Kaplan-Meier analysis, patients with high CXCR6 expression had significantly shorter overall survival than did those with low CXCR6 expression. The elevated co-expression levels of CXCL12/CXCR4 and CXCL16/CXCR6 in CIN and cervical carcinoma suggest a durative process in cervical carcinoma development. Moreover, CXCR6 may be useful as a biomarker and a valuable prognostic factor for cervical cancer.

[MicroRNA-16 regulates the proliferation, invasion and apoptosis of ovarian epithelial carcinoma cells in vitro].

OBJECTIVE: To study the role and mechanism of microRNA-16 (miR-16) in the proliferation, invasion and apoptosis of ovarian epithelial carcinoma cells in vitro. METHODS: The SKOV-3 cells were transfected with miR-16 mimics or negative control RNA (NC) by lipofectamine 2000. The expression of miR-16 was detected by real-time reverse transcription (RT)-PCR in SKOV-3 cells, and western blot was used to detect the expression of vascular endothelial growth factor (VEGF), matrix metalloproteinase-2 (MMP-2) and bcl-2 protein. Methyl thiazolyl tetrazolium (MTT), 5-ethynyl-2'-deoxyuridine (EdU) and transwell assay were used to determine the proliferation and invasion abilities. And the rate of apoptotic cell was detected by flow cytometry method. RESULTS: (1) The expression level of miR-16 in the transfection cells group was significantly higher than that in NC group (125.93 ± 15.30 versus 0.78 ± 0.16, P < 0.01). (2) The relative expression level of VEGF protein in transfection cells, NC and blank control group was 0.58 ± 0.05, 1.22 ± 0.03, 1.20 ± 0.03, MMP-2 protein was 0.63 ± 0.03, 1.16 ± 0.03, 1.21 ± 0.03, and bcl-2 protein 0.52 ± 0.03, 1.19 ± 0.05, 1.28 ± 0.06, respectively. The level of VEGF, MMP-2 and bcl-2 protein in the transfection group were lower than those in other control groups, and there were significantly differences among them (all P < 0.01). (3) After transfected 4 days, the inhibition rate of cell proliferation in the transfection group was dramatically higher than that in NC group [(37.2 ± 6.2)% versus (3.6 ± 3.2)%, P = 0.001]. (4) The percentage rate of proliferative cells in the transfection, NC and blank control group was (12.3 ± 0.8)%, (23.4 ± 1.8)%, (31.1 ± 4.9)%. And it was lower in the transfection group (P < 0.05). (5) Decreased cells via the transwell member in the transfection group (6 ± 3) were detected as compared with NC group (40 ± 9) and blank control group (48 ± 8, P < 0.01). (6) Twenty-four hours after cultured in serum starvation and hypoxia, the rate of the viable and late apoptotic cells in the transfection group were significantly higher than those in NC group and blank control group [the rate of viable apoptotic cell was (16.9 ± 2.1)%, (10.3 ± 1.7)% and (9.0 ± 0.8)% respectively, P < 0.01; the rate of late apoptotic cell was (13.4 ± 3.3)%, (3.2 ± 1.8)% and (0.7 ± 0.6)% respectively, P < 0.01]. After cultured 48 hours, total apoptotic cells in the transfection group was significantly more than those in other groups (P < 0.01). CONCLUSION: miR-16 might inhibit the proliferation, invasion of ovarian epithelial carcinoma cells and enhance their sensitivity to apoptotic stimuli via downregulation of the expression of VEGF, MMP-2 and bcl-2 protein.

[Role of microRNA-21 in the proliferation and apoptosis of ovarian epithelial carcinoma cells].

OBJECTIVE: To investigate the role and mechanism of microRNA-21(miR-21) in the proliferation and apoptosis of ovarian epithelial carcinoma cells. METHODS: A short-hairpin RNA specifically targeting miR-21 plasmid was constructed, and the recombinant was identified by restriction endonuclease analysis and DNA sequencing. Three experimental groups were included, transfection group (transfected with pSIREN-miR-21), negative control group (transfected with pSIREN-miR-21-neg) and blank control group (without transfection plasmid). The expression of miR-21 was detected by stem-loop real-time reverse transcription (RT)-PCR in OVCAR3 cells, and western blot was used to detect the expression of programmed cell death 4 (PDCD4) protein. Tethyl thiazolyl tetrazolium(MTT) and flow cytometry method were used respectively. RESULTS: Recombinant plasmid (pSIREN-miR-21) was constructed successfully and identified by restriction endonuclease analysis and DNA sequencing. The relative expression level of miR-21 in cells transfection, negative control and blank control group was 0.26 ± 0.08, 1.26 ± 0.21 and 1.00 respectively. The level of miR-21 in the cells in transfection group was significantly lower than those in the negative control and blank control group (P < 0.01). The gray scale of PDCD4 protein was 1443 ± 33, 858 ± 19 and 846 ± 16 in the transfection group, negative control and blank control group respectively. The value of PDCD4 in transfection group was higher than other control groups, and there were significantly difference among them(P < 0.01). Moreover, the optical density of the cells in transfection group was 0.661 ± 0.015, significantly lower than those in two control groups (0.848 ± 0.150 for negative control, 0.935 ± 0.133 for blank control, P < 0.01). Forty-eight hours after transfection, the rate of viable apoptotic cell was significantly higher than negative control and blank control group [(25.821 ± 0.763)% vs. (0.010 ± 0.003)% vs. (0.238 ± 0.023)%; P < 0.01]; 72 hours after transfection, the rates of viable apoptotic cell and necrotic cell were all higher than the two control groups [the rate of viable apoptotic cell was (30.480 ± 0.821)%, (7.792 ± 0.312)% and (7.033 ± 0.257)% respectively (P < 0.01); the rate of necrotic cell was (3.558 ± 0.211)%, (1.557 ± 0.067)% and (1.049 ± 0.028)%, respectively (P < 0.01)]. CONCLUSION: miR-21 might play an important role in the proliferation and apoptosis of ovarian epithelial carcinoma cells through negatively control the expression of PDCD4.

Chemokine axes CXCL12/CXCR4 and CXCL16/CXCR6 correlate with lymph node metastasis in epithelial ovarian carcinoma.

Recent evidence suggests that the chemokine axis of CXC chemokine ligand-12 and its receptor CXC chemokine receptor-4 (CXCL12/CXCR4) is highly expressed in gynecological tumors and the axis of CXC chemokine ligand-16 and CXC chemokine receptor-6 (CXCL16/CXCR6) is overexpressed in inflammation-associated tumors. This study aimed to determine the relationship between CXCL12/CXCR4, CXCL16/CXCR6 and ovarian carcinoma's clinicopathologic features and prognosis. Accordingly, the expression of these proteins in ovarian tissues was detected by tissue microarray and immunohistochemistry. The expressions of CXCL12/CXCR4 and CXCL16/CXCR6 were significantly higher in epithelial ovarian carcinomas than in normal epithelial ovarian tissues or benign epithelial ovarian tumors. The expression of chemokines CXCL12 and CXCL16 were positively correlated with their receptors CXCR4 and CXCR6 in ovarian carcinoma, respectively (r = 0.300, P < 0.05; r = 0.395, P < 0.05). Moreover, the expression of CXCL12 was related to the occurrence of ascites (Χ² = 4.76, P < 0.05), the expression of CXCR4 was significantly related to lymph node metastasis (Χ(2) = 4.37, P < 0.05), the expression of CXCR6 was significantly related to lymph node metastasis (Χ² = 7.43, P < 0.05) and histological type (Χ² = 33.48, P < 0.05). In univariate analysis, the expression of CXCR4 and CXCL16 significantly correlated with reduced median survival (Χ² = 4.67, P < 0.05; Χ² = 4.48, P < 0.05). Therefore, we conclude that the chemokine axes CXCL12/CXCR4 and CXCL16/CXCR6 may play important roles in the growth, proliferation, invasion, and metastasis of epithelial ovarian carcinoma.

Factors associated with attitudes toward tobacco control policy in public places among adults in three counties of China / 中华流行病学杂志

Objective To investigate the attitudes of urban and rural community members toward total banning on smoking in public places and to explore the factors associated with these attitudes, in three counties/cities in China. Methods A cross-sectional study was conducted in three counties/cities in 2004, including Xin' an county of Henan province, Anyi county of Jiangxi province, and Mianzhu city of Sichuan province. A total of 5642 residents at age of 18-69 years old were interviewed face-to-face with a uniform questionnaire by locally-trained interviewers, through a random three-stage stratified sampling in each county. Factors were assumed to be associated with attitudes, using chi-square test in univariate analysis and non-conditional logistic regression model in multivariate analysis. Results The prevalence of current smoking among respondents was 44.3%. 80.7% (1379/1709) of the current smokers reported smoking often or sometimes in public places. Only 9.6% (479/4983) of the respondents reported that their indoor workplaces had totally banned on smoking. 43.5% of the respondents supported a total smoking ban strategy in pubic venues. The results of multivariate logistic regression model showed that eight factors were significantly associated with support for the total smoking ban in public places included region, residency, age, gender, education, smoking status, awareness about passive smoking hazards, and hearing of any message on tobacco control through media differences of rates regarding the factors as: residents in urban to rural areas (OR=1.29), elderly to youngsters (30-49 vs. 18-29, OR=1.46; 50-69 vs. 18-29, OR=1.71), female to male(OR=1.27), high-educated to less-educated ones, quitters to current smokers (OR=1.90), nonsmokers to current smokers (OR=2.01). Those who know messages on health hazards of passive smoking (OR=2.26), or heard of message on tobacco control through media (OR= 1.43). Conclusion Results from our study revealed that a thorough smoke-flee policy in public places should be developed and implemented in these three counties/cities.

Exploring barriers to implementation of smoking policies: a qualitative study on health professionals from three county-level hospitals / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>This study was to identify factors limiting the implementation of smoking policies in county-level hospitals.</p><p><b>METHODS</b>We conducted qualitative interviews (17 focus groups discussions and 6 one-to-one in depth interviews) involving 103 health professionals from three target county-level hospitals. A combination of purposive and convenience sampling was used to recruit subjects and gain a broad range of perspectives on issues emerging from ongoing data-analysis until data saturation occurred. The transcripts were analyzed for themes and key points.</p><p><b>RESULTS</b>The main themes that emerged suggested that both smokers and non-smokers viewed smoking very negatively. However, it was clear that, underlying this acceptance of the health risks of smoking, there was a wide range of beliefs. Most of the health professionals pointed out that, as smoking was legal, addictive, and influenced by social norms, currently it was almost unrealistic to expect all smokers to give up smoking or not to smoke in the hospitals. Furthermore, they were concerned about the potentially detrimental effects of providing counseling advice to all smokers on the interpersonal relationship among colleagues or between doctors and patients. In addition, low level of employee participation influenced the sustainable implementation of smoking policies.</p><p><b>CONCLUSIONS</b>Simply being aware of the health risks about smoking did not necessarily result in successful implementation of the smoking policies. Application of comprehensive intervention strategies such as implementing smoking policies in public places at the county level, creating supportive environments, promoting community participation, and conducting health education, may be more effective.</p>

The current status of passive smoking in Chinese families and associated factors / 中华预防医学杂志

<p><b>OBJECTIVE</b>To understand the prevalence of passive smoking in Chinese families and discuss its associated factors, as to providing scientific evidence for establishing tobacco control measures in China.</p><p><b>METHOD</b>Cross-sectional survey: from June to September, 2004, we randomly selected six counties in three different provinces ( Mianzhu and Xichong of Sichuan Province; Anyi and Hukou of Jiangxi Province; Xinan and Yanshi of Henan Province) and performed face-to-face questionnaire survey on citizens between 18 and 69 years old. All the data were double independently input by professional data entry company to ensure data accuracy. The prevalence of home passive smoking exposure in families with different demographic characteristics was described by using prevalence, and the possible correlated factors of home passive smoking exposure as independent variables, multiple factors were analyzed using Logistic Stepwise Regression Analysis method.</p><p><b>RESULTS</b>The analysis on 8142 nonsmokers revealed that the rate of passive smoking was 28.42%, with 27.38% of male and 28.93% of female suffering from passive smoking. All 87.19% of the smokers would smoke in front of their families. As many as 42.14% of the nonsmokers would offer cigarettes to their guests, while about 46.82% of the nonsmokers would suggest smokers to smoke outdoor. Home restriction on tobacco was extremely rare and only 6.33% of all the families completely forbade smoking. Multivariate logistic regression analysis of non-conditions revealed that, there was a lower level of involuntary tobacco smoke exposure in female, older age group, lower education level, divorced, or widowed families. There was no difference in involuntary tobacco smoke exposure between town dwellers and county dwellers, but such difference did exist in different districts.</p><p><b>CONCLUSION</b>The three provinces under investigation should have severe involuntary tobacco smoking exposure. Gender, age, literacy level, occupation and region should be all factors that influence the status of involuntary tobacco smoking exposure in different families. There is a high percentile that smokers would smoke in front of their families and kids and a relative low pressure against smoking from nonsmokers. Cigarette offering is very prevalence. The knowledge and attitude about passive smoking should be separated from the situation of passive smoking exposure.</p>

Passive smoking in China: contributing factors and areas for future interventions / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To reduce tobacco consumption and exposure to passive smoking in China.</p><p><b>METHODS</b>Discussion consisting of 80 focus groups and 35 interviews were held in three rural intervention counties of Jiangxi, Henan, and Sichuan Provinces. Participants came from hospitals, schools, rural areas, and urban areas.</p><p><b>RESULTS</b>Tobacco use and exposure to passive smoking were widely prevalent in the investigated schools, hospitals, county towns, and rural areas. Knowledge of the risks for passive smoking on health is lacking, especially in rural areas. Barriers to the control of tobacco use in public places include reluctance of administrators to implement tobacco control policies, lack of consistent policies, difficulties with regulations and enforcement, and reluctance of non-smokers to exercise their right to clean air.</p><p><b>CONCLUSION</b>To curb the current tobacco epidemic in China, tobacco control efforts must focus on reducing exposure to passive smoking. A strategy should be formulated to reduce the factors that contribute to tobacco use and exposure to passive smoking.</p>

[Effect of small interference RNA on E6, E7 mRNA of human papillomavirus type-18 in cervical cancer cells].

OBJECTIVE: To study the effect of small interference RNA on E6, E7 mRNA of human papillomavirus type-18 (HPV18) in cervical cancer cells. METHODS: The specific HPV18E6 and E7 siRNA synthesized by in virtro transcription. The cell activity was detected by methyl thiazolyl tetrazolium (MTT) assay to determine the most suitable concentration, then cells were transfected into HPV18 cervical cancer cells by oligofectamine. HPV18E6, E7 mRNA level was examined by semi-quantitative RT-PCR, and the cell cycles were analyzed by flow cytometry. RESULTS: The activity of the HeLa cells had an obvious suppression after transfection to 0.57 +/- 0.05 and 0.62 +/- 0.04 compared with the contrast of 0.87 +/- 0.05. The expression of HPV18E6 mRNA level was 0.63 +/- 0.04 before transfection, decreased to 0.53 +/- 0.04, 0.46 +/- 0.02, 0.56 +/- 0.03 evidently after transfection at 24, 48, 72 hours. The expression of HPV18E7 mRNA level was 0.66 +/- 0.03 before transfection and decreased to 0.60 +/- 0.05, 0.52 +/- 0.04, 0.59 +/- 0.02 after transfection at 24, 48, 72 hours. The number of cells in G(2) phase was increased after HPV18E6 siRNA transfection from (1.4 +/- 1.2)% to (66.9 +/- 3.5)%, S phase was declined from (39.4 +/- 0.4)% to (0 +/- 5.5)%, and they were (47.2 +/- 0.5)% and (5.6 +/- 4.2)% in E7 group. CONCLUSION: RNAi exists in HeLa cells, and its effect has specificity.

[Expression of SLP-2 mRNA in endometrial cancer and its significance].

OBJECTIVE: To characterize the differential expression of SLP-2 in endometrial cancer, and to study the effect of human SLP-2 gene on human endometrial cancer cell line. METHODS: The expression of SLP-2 gene in 32 cases of endometrial cancer and 28 cases of normal endometrial tissues was examined by semi-quantitative RT-PCR. Eukaryotic expression vectors of sense and antisense SLP-2 were constructed and transfected into HEC-1B cell line using lipofectamine 2000 respectively. The morphological changes of the cell were observed by phase contrast microscopy. The cell growth was detected by methyl thiazolyl tetrazolium (MTT) assay, and the cell cycles were analyzed by flow cytometry. RESULTS: The expression of SLP-2 mRNA in endometrial cancer tissues was higher than that in normal endometrial tissues (1.6 +/- 0.7 vs 0.7 +/- 0.3, P < 0.05). Sense and antisense human SLP-2 constructs were transfected into HEC-1B cell line respectively. After being transfected with sense SLP-2, the expression of SLP-2 mRNA in HEC-1B cell line was increased by about 2.4 times that of the control group, the cell growth was accelerated, and the number of cells in G(1) phase was decreased by 12.5%, S phase was increased by 8.0%. After being transfected with antisense SLP-2, the expression of SLP-2 mRNA was declined 50%. The transfected cells showed slower growth, and the number of cells in G(1) phase was significantly increased by 10.5%, and S phase was declined by 9.8%. CONCLUSION: SLP-2 mRNA shows up-regulation in endometrial cancer tissues, and it may have some relationship with carcinogenesis of endometrial cancer.

[In vivo reversal of multidrug resistance by transduction of human tumor necrosis factor-alpha into drug resistant cell line of choriocarcinoma].

OBJECTIVE: To investigate in vivo reversal of multidrug resistance and biological properties of a drug resistant cell line of choriocarcinoma transduced by human tumor necrosis factor-alpha (TNF-alpha) gene via the establishment of its animal model. METHODS: Choriocarcinoma cell line JEG-3, drug-resistant choriocarcinoma cell line JEG-3/VP2, and human TNF-alpha-transduced drug-resistant choriocarcinoma cell line JEG-3/VP2/TNF-alpha were injected subcutaneously in the neck of nude mices. Tumor size and weight were routinely measured, tumor histological structure was observed and its chemosensitivity was tested. Expression of multidrug resistance (MDR1) mRNA was investigated using reverse transcription-polymerase chain reaction (RT-PCR) and P-glycoprotein (P-gp) expression was determined by immunohistochemistry with the monoclonal antibody MDR1. RESULTS: The rate of inoculation for all three tested cell lines was 100%, the latent period was 10 to 14 days. Tumor growth rates and weights were significantly different among three cell lines (P < 0.05), with the lowest in JEG-3/VP2/TNF-alpha cell line. Tumor inhibition rate after treatment with etopside (VP-16) was significantly higher in JEG-3/VP2/TNF-alpha (41.0% - 42.5%) (P < 0.05), compared with JEG-3/VP2 (24.3% - 28%), and similar to JEG-3 (46.7% -47.7%). Transduction and expression of human TNF-alpha in drug-resistant choriocarcinoma cell line JEG-3/VP2 was found to reverse MDR1 on the mRNA and P-gp levels. CONCLUSION: Transduction and expression of human TNF-alpha in drug-resistant choriocarcinoma cell line JEG-3/VP2 can reverse expressions of MDR1 mRNA and P-gp, enhance the susceptibility of the JEG-3/VP2 to the cytotoxic drugs, and lower its tumorigenesis.