Difference between revisions of "Team:Fudan/HCC"

Line 66: Line 66:
 
</p>
 
</p>
 
   </div>
 
   </div>
 +
  
 
   <div class="block">
 
   <div class="block">
−
     <h class="highlight-title silver-green">Method</h>
+
     <h class="highlight-title silver-green">Wilderness</h>
 
     <p>
 
     <p>
−
       8 plasmids (Positive Control, Negative Control, Test Device 1, Test Device 2, Test Device 3, Test Device 4, Test Device 5, and Test Device 6) from Kit Plate 6 or Kit Plate 7 were transformed into E. coli DH5-alpha cells. After transformation and cultivation, the cells are ready for experiments through the provided protocol. Generally, plate reader is suggested as the measuring tool, but flow cytometer is also acceptable.<br />
+
       During 2012, 782,500 (male 554,400; female 228,100) new cases of liver cancer and 745,500 (male 521,100, second leading cause of cancer death among males; female 224,500) deaths in total occurred around the world. Meanwhile, China alone accounted for around 50% of the total number of cases and deaths 1. China’s liver cancer cases are still rising sharply. Later statistics reported that new cases and death cases climb to 466,100 and 422, 100 around China during 20152. According to World Health Organization (WHO), the primary liver cancers can be classified into primary hepatocellular carcinoma (HCC), cholangiocarcinoma (ICC), and combined hepatocellular and cholangiocarcinoma (cHCC-CC)3, among which HCC is the most common occurring, making up 80% of cases4. As the sixth most frequent and the third most lethal tumor5, HCC brings great harm to the Chinese health. How to improve HCC treatment and raise patient survival rate has become an urgent social problem nowadays.  
 
     </p>
 
     </p>
 +
 
   </div>
 
   </div>
  
−
  <div class="block">
+
<div class="block">
−
     <h class="highlight-title silver-green">Protocol</h>
+
     <h class="highlight-title silver-green">Wilderness</h>
 
     <p>
 
     <p>
−
       Transformation Protocol:<br />
+
       During 2012, 782,500 (male 554,400; female 228,100) new cases of liver cancer and 745,500 (male 521,100, second leading cause of cancer death among males; female 224,500) deaths in total occurred around the world. Meanwhile, China alone accounted for around 50% of the total number of cases and deaths 1. China’s liver cancer cases are still rising sharply. Later statistics reported that new cases and death cases climb to 466,100 and 422, 100 around China during 20152. According to World Health Organization (WHO), the primary liver cancers can be classified into primary hepatocellular carcinoma (HCC), cholangiocarcinoma (ICC), and combined hepatocellular and cholangiocarcinoma (cHCC-CC)3, among which HCC is the most common occurring, making up 80% of cases4. As the sixth most frequent and the third most lethal tumor5, HCC brings great harm to the Chinese health. How to improve HCC treatment and raise patient survival rate has become an urgent social problem nowadays.  
−
      <a href="http://parts.igem.org/Help:Protocols/Transformation">http://parts.igem.org/Help:Protocols/Transformation</a><br />
+
−
    </p>
+
−
    <p>
+
−
      Plate Reader Protocol: <br />
+
−
      <a href="https://2017.igem.org/Competition/InterLab_Study/Plate_Reader">https://2017.igem.org/Competition/InterLab_Study/Plate_Reader</a><br />
+
−
    </p>
+
−
    <p>
+
−
      Flow Cytometer Protocol: <br />
+
−
      <a href="https://2017.igem.org/Competition/InterLab_Study/Flow_Cytometry">https://2017.igem.org/Competition/InterLab_Study/Flow_Cytometry</a><br />
+
 
     </p>
 
     </p>
 +
<p>
 +
 +
</p>
 +
<p>
 +
Behind the cold data is an icy fact: every single one counts for the death toll is a living world for someone else, leaving behind widows, widowers and orphans. As a creature with humanity and compassion, we feel the pain deeply; these torture our souls constantly. The torpor is still wandering the mist, while those who suffered had come to their conscious and step forward, directly into the unknown wilderness.
 +
</p>
 
   </div>
 
   </div>
  
−
  <div class="block">
 
−
    <h class="highlight-title silver-green">Results</h>
 
−
    <ul>
 
−
      <li>
 
−
        <p>OD 600 reference point</p>
 
−
        <p>Using OD 600 and H2O to generate the conversion factor for the transformation later. The average of OD600 is 0.04725; the correction factor (OD600/ABS600) is 5.6666667</p>
 
−
        <p style="text-align:center;"><img src="https://static.igem.org/mediawiki/2017/0/05/T--Fudan--Interlab--figure1.png" /></p>
 
−
        <p style="text-align:center;">Figure1, OD 600 reference point</p>
 
−
      </li>
 
−
      <li>
 
−
        <p>Fluorescein standard curve</p>
 
−
        <p>Dilution serious of fluorescein were prepared and measured in a 96 well plate. A standard curve is generated to correct the cell based readings to an equivalent fluorescein concentration.</p>
 
−
        <p style="text-align:center;"><img src="https://static.igem.org/mediawiki/2017/2/2d/T--Fudan--InterLab--figure2.png" /></p>
 
−
        <p style="text-align:center;"><img src="https://static.igem.org/mediawiki/2017/e/ec/T--Fudan--InterLab--figure3.png" /></p>
 
−
        <p style="text-align:center;">Figure 2: Fluorescein standard curve</p>
 
−
      </li>
 
−
      <li>
 
−
        <p>Fluorescein standard curve</p>
 
−
        <p>Dilution serious of fluorescein were prepared and measured in a 96 well plate. A standard curve is generated to correct the cell based readings to an equivalent fluorescein concentration.</p>
 
−
        <p style="text-align:center;"><img img width="80%" src="https://static.igem.org/mediawiki/2017/1/1a/T--Fudan--InterLab--figure4.png" /></p>
 
−
        <p style="text-align:center;">Figure 3: Raw OD600 with background</p>
 
−
        <p>Show the growing curve of different cells indirectlly. The test devices 2,3,5,6 show a higher growing comparing to devices 1 and 4.The negative control group shows a high growing comparing to the positive control.</p>
 
−
        <p style="text-align:center;"><img width="80%" src="https://static.igem.org/mediawiki/2017/8/8d/T--Fudan--InterLab--figure5.png" /></p>
 
−
        <p style="text-align:center;">Figure 4: Fluorescence – background</p>
 
−
        <p>Test devices 2 and 4 show high fluorescence intensity alone with positive control group. Test devices 1 and 5 show a modest fluorescence intensity, while devices 3 and 6 barely show low fluorescence intensity alone with the negative control group.</p>
 
−
        <p style="text-align:center;"><img width="80%" src="https://static.igem.org/mediawiki/2017/6/63/T--Fudan--InterLab--figure6.png" /></p>
 
−
        <p style="text-align:center;">Figure 5: Cell measurement</p>
 
−
      </li>
 
−
    </ul>
 
−
    <p>
 
−
      <a href="https://static.igem.org/mediawiki/2017/2/20/T--Fudan--Interlab_Data.xlsx">Click here to see the complete data.</a><br />
 
−
    </p>
 
−
  </div>
 
−
 
 
−
  <div class="block">
 
−
    <h class="highlight-title silver-green">Discussions</h>
 
−
    <p>
 
−
      In figure 3, alone with the negative control group, several test devices show a high OD600 value, suggesting a relatively unaffected growth, while some devices like test devices 1 and 4 have a low grow rate. Possible explanation for this is that a high expression of GFP as an unnecessary protein will influence the rate of cell growth. Growth peak probably will show up after h6.<br />
 
−
    </p>
 
−
    <p>
 
−
      In figure 4, highest Fluorescence was obtained from device 2, closely followed by positive control and test device 4. Test device 1 and 5 show a modest fluorescence intensity, while test devices 3 and 6 barely have any fluorescence signal as well as the negative group.<br />
 
−
    </p>
 
−
    <p>
 
−
      Combined with figure 1, we draw the figure 3, showing the expression of GFP on average.<br />
 
−
    </p>
 
−
    <p>
 
−
      For test device 1, it had a rather high average expression at the start, but decreased later. This may suggest a constitutive high expression of GFP affects the growth and cell activity, since the total Fl signal was depended on both average expression strength and cell density.<br />
 
−
    </p>
 
−
    <p>
 
−
      For test device 2, though a high total signal intensity was achieved, the average expression was rather low.<br />
 
−
    </p>
 
−
    <p>
 
−
      For test device 3 and 6, we found a similar results comparing with the negative group, this may result from rather low expressions close to zero and unaffected growth.<br />
 
−
    </p>
 
−
  </div>
 
−
 
 
−
  <div class="block">
 
−
    <h class="highlight-title silver-green">Conclusions</h>
 
−
    <p>
 
−
      Experiment achieved a rather normal result, showing the robustness of the protocol alone with standardized cells and circuit. The Interlab study do offer a platform for global communication, which is of vital importance in synthetic biology in the future.<br />
 
−
    </p>
 
−
  </div>
 
−
 
 
 
</div>
 
</div>
  

Revision as of 06:26, 29 October 2017
Prologue

When the aroma of wine fills the bistro and bard's harp rings jubilant; Or when starlight shines above the earth, declaring the busy day coming to an end; Storybooks are going to be opened; words will come to live again. All sufferings turned to gold, and all whishes became accomplishments. It was a piece of history that us human struggling with cancer. "SwordS", as they called the name.

Revelation

Hepatocellular carcinoma, HCC is a kind of malignant tumor occurred in liver that has a universal feature—the presence of structural and numeric chromosomal abnormalities indicative of genomic instability. In most cases, it develops from small-cell, high-grade dysplastic nodules in cirrhotic livers. On histologic examination, HCCs range from well-differentiated lesions that reproduce hepatocytes arranged in cords, trabeculae or glandular patterns to poorly differentiated lesions, often composed of large, multinucleate anaplastic giant cells. It may be unifocal, multifocal or diffusely infiltrative in pathologic samples. HCC has a strong propensity for vascular invasion and recapitulates normal liver architecture to varying degrees. In summary, HCC is a serious malignant tumor with high heterogeneity. (Adapt from Robbins Basic Pathology 9th)

Wilderness

During 2012, 782,500 (male 554,400; female 228,100) new cases of liver cancer and 745,500 (male 521,100, second leading cause of cancer death among males; female 224,500) deaths in total occurred around the world. Meanwhile, China alone accounted for around 50% of the total number of cases and deaths 1. China’s liver cancer cases are still rising sharply. Later statistics reported that new cases and death cases climb to 466,100 and 422, 100 around China during 20152. According to World Health Organization (WHO), the primary liver cancers can be classified into primary hepatocellular carcinoma (HCC), cholangiocarcinoma (ICC), and combined hepatocellular and cholangiocarcinoma (cHCC-CC)3, among which HCC is the most common occurring, making up 80% of cases4. As the sixth most frequent and the third most lethal tumor5, HCC brings great harm to the Chinese health. How to improve HCC treatment and raise patient survival rate has become an urgent social problem nowadays.

Wilderness

During 2012, 782,500 (male 554,400; female 228,100) new cases of liver cancer and 745,500 (male 521,100, second leading cause of cancer death among males; female 224,500) deaths in total occurred around the world. Meanwhile, China alone accounted for around 50% of the total number of cases and deaths 1. China’s liver cancer cases are still rising sharply. Later statistics reported that new cases and death cases climb to 466,100 and 422, 100 around China during 20152. According to World Health Organization (WHO), the primary liver cancers can be classified into primary hepatocellular carcinoma (HCC), cholangiocarcinoma (ICC), and combined hepatocellular and cholangiocarcinoma (cHCC-CC)3, among which HCC is the most common occurring, making up 80% of cases4. As the sixth most frequent and the third most lethal tumor5, HCC brings great harm to the Chinese health. How to improve HCC treatment and raise patient survival rate has become an urgent social problem nowadays.

Behind the cold data is an icy fact: every single one counts for the death toll is a living world for someone else, leaving behind widows, widowers and orphans. As a creature with humanity and compassion, we feel the pain deeply; these torture our souls constantly. The torpor is still wandering the mist, while those who suffered had come to their conscious and step forward, directly into the unknown wilderness.