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The Delta Lotto System is our free system for choosing lotterynumbers based on a statistical study of the mathematical differencesbetween adjacent numbers. We believe it to be the best lotto system available. Below, as you will see, we can actually pick for a 50-digit lotto game by choosing only numbers betweek 1 and 15!

We will be choosing six lotto numbers by firstchoosingsix delta numbersbetween 1 and 15, following all the rules of our system. All thefollowing examples assume a 6-digit, 1 to 50 lotto game. Theserules will change somewhat for games which are too differentfrom this - for Powerball or Megamillions, for example, 18 to 22 wouldbe the top delta number. Use the AnalysisLottosoftware, or our free lottocomputer to experiment with different games. Additional helpcan be also found on our DeltaLotto DVD.

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Start by choosing one very low number. Over 60% ofthetime, ONE is going tobe at least one of the winning delta numbers. So, if you feel goodabout it, make the number ONEyour first choice. If you don't pick ONE,choose another very low number like two or three.

The last two numbers should probably be higher than8but not higher than 15. Less than half the time, a winning number mayhave a delta higher than 15. Go ahead and make one of these numbershigher than 15 (but less than 25) if it feels right... but my advice isto mostly stick to numbers 15 and below, where the odds of picking theright numbers are more favorable.

What's the highest number allowed in your lottogame?Add up all the delta numbers you picked and make sure its not higherthan this number. If this sum is within about FIFTEENof the highest number, it has the best chance of being a winner. Goahead and change some of the numbers if you need to to make the sumcome out right. Remember, its OK to have numbers repeat in yoursequence.

Perform the addition sequence below to discover theactuallotto number you'll be playing. HINT: pay attention to when we sayLOTTO number and DELTA number. If you have any problemfollowingthis, go back to our first page and watch the video there closely.

I have made a research and a review of around 10 different lotto systems and none of them have been worth it. They have always big promises how you could become a millionaire or at least win more often in the lottery.

It is a software that you can download to your computer and its purpose is to show you the winning lotto numbers. They have lots of testimonials on their colorful circus page who claim that they have doubled, tripled, quadrupled their money. Or even much more.

As I mentioned earlier, I have never seen a lotto program that would really work and make you more money. But I know many lotto systems that take money from your pocket twice. First, when you buy a lotto system you need to pay the price, then you pay again when you buy a lottery ticket. In the end, you will end up spending much more money than you had planned.

If you are interested in making money online, I recommend going away from lotto systems and taking a look at better opportunities. My #1 recommendation is Wealthy Affiliate because they provide proven methods how you can make a living online.

I find these lotto systems quite hilarious because they claim with a serious face that you could beat the lottery with their product. As I mentioned in the video, if that would be true, these guys would already be billionaires.

Therefore, I think lotto systems are just useless. Waste of time and money. What do you think Dave? What is your argument why Lotto software or system would be worth it if it still loses money in the long run?

Hepatocellular carcinoma (HCC) remains a major public health problemworldwide. The identification of effective chemotherapeutic targets foradvanced HCC patients is urgently required. In this study, we investigatedthe role of protein phosphatase 2A-B55]delta] subunit (PP2A-B55]delta],encoded by the

, were performed to explore the function of B55]delta] in cisplatin (cDDP)chemotherapy of HCC. Bioinformatics prediction, luciferase reporter assays,qRT-PCR, WB, and cell cycle analyses were used to reveal the regulatoryrelationship between microRNA-133b (miR-133b) and

expression was down-regulated in both HCC tumors and HCC cell lines.Treatment with cDDP increased the amount of B55]delta] protein. Artificiallyincreasing the expression of B55]delta] counteracted cyclin-dependent kinase1 activation, modulated transitions of the cell cycle, and increased thesuppressive effect of cDDP on cell migration, colony formation, apoptosis,and proliferation

PP2A-B55]delta], regulated by miR-133b, enhances the sensitivity of HCC tocDDP chemotherapy. Our data indicate that PP2A-B55]delta] might be a noveland attractive target for increasing chemotherapy sensitivity of HCC.

In recent years, a growing number of studies have shown that proteinphosphatase 2A (PP2A) is an important tumor suppressor. As a crucial memberof the serine/threonine protein phosphatase family widely expressed ineukaryotic cells, PP2A is involved in the regulation of the signaltransduction, cell cycle, proliferation, differentiation, apoptosis, andother processes [3]. PP2A-B55]delta] subunit, encoded by the PPP2R2D gene, is one of four isoforms ([alpha], [beta], , and ]delta]) of thePP2A B55 regulatory subunit family [4]. The interaction between B55]delta]and cyclin-dependent kinase 1 (CDK1) is reported to play a critical role incell cycle progression [5]. However, it is still unclear whether B55]delta]enhances chemotherapy sensitivity of HCC cells by regulating the cell cycle.

As a key drug in the treatment of advanced HCC [12], cisplatin (cDDP) wasselected as a representative chemotherapeutic drug of HCC in this study. Weexplored the role of PP2A-B55]delta] both in regulating the cell cycle,migration, colony formation, apoptosis, and proliferation of humanhepatoblastoma HepG2 cells and in tumor growth in xenograft mice in thepresence of cDDP, and we investigated the details of the microRNA-133b(miR-133b)/PPP2R2D signaling pathway. We concluded that PP2A-B55]delta], under the regulationof miR-133b, could serve as a promising target for increasing chemotherapysensitivity of HCC.

The short hairpin RNA (shRNA) targeting the PPP2R2D mRNA sequence (GenBank Accession No. NM_018461.4) was designed using theGenetic Perturbation Platform ( ).The sense and antisense oligonucleotides of sh2R2D (Additional file 1: Table S1) were annealed and then ligated into lentiviralvector pLKO.1-puro (a kind gift from Dr. Wen Chen, Sun Yat-sen University,China) to construct the pLKO.1-sh2R2D recombinant plasmid. The corresponding control plasmid was pLKO.1 bearingshRNA targeting green fluorescent protein (pLKO.1-shGFP) [13]. The lentiviral plasmid (pLKO.1-shGFP or pLKO.1-sh2R2D), packaging plasmid (pCMV-delta 8.9), and envelope plasmid (VSVG) wereco-transfected into HEK-293T cells using X-tremeGENE HP DNA transfectionreagent (Roche, IN, USA). Lentiviruses packaged in HEK-293T cells werepurified and then introduced into HepG2 cells using polybrene (Sigma, MO,USA). The established cell lines (HepG2-shGFP and HepG2-sh2R2D) were screened with 0.6 [mu]g/ml puromycin.

Fig. 1: PPP2R2D is down-regulated in HCC. a PPP2R2D expression levels in two cohorts of HCC patients [14, 15]. The mRNAmicroarray data were obtained from GEO database (Accession No.: GSE59259 andGSE6764, *P < 0.05). b Kaplan-Meier analysis showing overall survival of HCC patients [16] with twodifferent levels of PPP2R2D expression (Accession No.: GSE54236, PPP2R2D expression was treated as a binary variable divided into high or lowexpression according to the median, Log-rank, *P < 0.05). c PPP2R2D mRNA and (d) B55]delta] protein levels were evaluated in normal hepatic cell lines andHCC cell lines. The data shown in the bar graph are the mean [+ or -] SD ofthree independent experiments. *P < 0.01 as compared with L02 cells; # P < 0.01 as compared with QSG7701 cells [see PDF for image]

We further evaluate PPP2R2D mRNA and B55]delta] protein in two normal hepatic cell lines and six HCCcell lines by qRT-PCR and WB. The data showed that the levels of PPP2R2D mRNA and B55]delta] protein were significantly suppressed in all HCC celllines compared with normal hepatic cell lines (Fig. 1c-d). Taken together,the expression of PPP2R2D was down-regulated in HCC tissues and cells, indicating that PPP2R2D might act as a tumor suppressor in the chemotherapy of HCC.

In order to explore the functional role of PPP2R2D in the chemotherapy of HCC, HepG2 cells were chosen as a sensitive modelbecause of its lower expression of PPP2R2D compared to other HCC cell lines (Fig. 1c-d). We used cDDP, an acceptedanti-tumor agent [17], investigating B55]delta] expression in HepG2 cells inresponse to chemotherapy. PP2A activity was increased by cDDP in aconcentration- and time-dependent manner (Fig. 2a). As shown in Fig. 2b, cDDPsignificantly up-regulated the mRNA level of PPP2R2D and PPP2R2A, but not of PPP2R1A, PPP2CA, or PPP2R5C. cDDP increased only the expression of B55]delta] protein, not of the othersubunits of PP2A (Fig. 2c). Immunofluorescence analysis also revealed thatB55]delta] was more strongly expressed following cDDP treatment (Fig. 2d).Additionally, the protein levels of B55]delta] were increased in cDDP-treatedcells in a concentration- and time-dependent manner (Fig. 2e). Thus, ourfindings suggested that cDDP increased the amount of B55]delta] in HepG2cells.

Fig. 2: The expression of B55]delta] is increased by cDDP in HepG2 cells. a PP2A activity and (e) B55]delta] protein levels in HepG2 cells treated with 0, 1, 2.5, or 4[mu]g/ml cDDP for 12 h, or treated with 2.5 [mu]g/ml cDDP for 0, 6, 12, or 24h. b-d HepG2 cells were treated with or without 2.5 [mu]g/ml cDDP for 12 h. b The mRNA levels of PP2A subunits (PPP2R1A, PPP2CA, PPP2R2A, PPP2R2D and PPP2R5C). c The protein levels of PP2A-A[alpha], -C, -B55[alpha], -B55]delta], and-B56 subunits. d Immunofluorescence images of B55]delta] in HepG2 cells (x630 magnification;scale bar, 10 [mu]m). Data are presented as mean [+ or -] SD of threeindependent assays. *P < 0.01 as compared with cDDP-untreated group (Ctrl) [see PDF for image] e24fc04721