The randomization list was passed to the Hospital Pharmacy which supplied the active compounds and placebo, and which prepared sequentially numbered containers of the study pills. The principal clinical investigator enrolled participants, and assigned participants to interventions. The EC administration started with an initial dose of 100/10 mg ter in die, for the first week and then proceeded with the full dose of 200/ 20 mg t.i.d. In order to evaluate drug safety, the blood pressure was measured three times a day; both electrocardiography and echocardiography were recorded at baseline and every week. Body weight was measured in a standardised manner at start and at the end of treatment. During the treatment period all patients were fed a hypocaloric diet, and containing 20% proteins, 55% carbohydrates, 25% fat half of which was monounsaturated, and 35 g/day fibres. They were hospitalised, during the whole treatment period, at the metabolic unit of San Giuseppe Hospital-Istituto Auxologico Italiano at Piancavallo. After diet and drug period, the patients were transferred to the Department of Surgery for the bariatric surgery. The drug treatment was stopped the day before surgical intervention. Small biopsies of rectus abdominis were taken during surgery, immediately frozen in liquid nitrogen, and stored at 280uC for subsequent analysis. Primary clinical outcome was the change in resting metabolic rate, while the primary non-clinical outcome was the UCP3 gene expression in skeletal muscle. Secondary outcomes were changes in body weight and metabolic parameters, and drug safety. Because only a few small studies have been done in humans to investigate the thermogenic effects of EC, in the present study we examined the UCP3 expression in skeletal muscle of premenopausal morbidly obese females treated with either placebo or EC for 28 days. Our findings demonstrate that chronic treatment with EC increased the RMR, in contrast with placebo which decreased the RMR in obese patients. This result is consistent with a previous study showing that the decrease in 24- hour energy expenditure seen in the placebo group was 10% at day 1 and 13% at day 56, but was only 7% and 8% in the ECtreated group, and the weight-loss was not different in the two groups after 8-week treatment.

The integration not only changes the transcription pattern relevant for the dysregulated expression of the viral oncogenes, but also affects the expression of the host gene with virus genome integration. The integration alters the expression of host genes in integration sites, even if this occurs within the intron sequences. In our study, we identified a broad spectrum of cancerassociated genes in the integration sites and flanking sequence regions. Most of genes in the integration sites were associated with tumor development, and nineteen genes were strongly related to cervical cancer. Some of them act as tumor suppressors or oncogenes. Interestingly, most of them were not reported in previous documents. MiR-34a, an important tumor suppressor, is down-regulated in cervical cancer. It has been reported that oncoprotein E6 of HPV16 and HPV18 can inhibit the expression of tumorsuppressive miR-34a by destabilization of p53 and resulted in cell proliferation. The disruption of miR-34a gene might further interpret the phenomenon of reduced expression of miR-34a in cervical cancer. MSH2 is a DNA mismatch repair protein, and associated with DNA repair pathway. Decreased expression of MSH2 might be a risk factor in the early stage cervical cancer. ROCK2, an important signaling molecule, can promote cervical cancer metastasis by upregulating and activating the expression and function of moesin protein through RhoA/ ROCK2 pathway. Besides the cancer-associated genes, the genes in integration sites and flanking sequence regions might be also beneficial for viral genome integration. FANCM which is a DNA translocase and highly related to DNA replication regulates checkpoint signaling and replication fork progression. Other genes, such as COX6B1 is related to cell apoptosis and ESRRA also have been reported associated with cervical cancer. In addition, among 45 integration events, 13 events led to antisense transcription of the coding sequences, such as PRDX5, EBAG9 and CD28, etc. These integrations were generally deemed of no interest.