In an effort to get a better understanding of the mechanisms of biofield therapy, further cell studies have been conducted in July, August, September, and October. We have added a new cell line and expanded the outcomes evaluated. • We have added Panc-02, a murine pancreatic cell line and examined the following doses of biofield therapy: 5 minutes; 15 minutes; 30 minutes • We have also explored multiple exposure to 15 minute doses in Panc-1 cells, i.e. 15 min one day; 15 minute a day for 2 days; 15 minute a day for 3 days. We will carry out the similar study in Hep3B cells in November, 2021. • We have done further work measuring cell voltage and characterization of cell changes as it relates to cell cycle. • We have also added the BrDU proliferation assay. On September 27, we started the planned, 3 week, C-Myc driven mouse hepatocelluar carcinoma animal study. In brief: **Animal model**: The LAP-tTA/tet-off c-Myc model is generated by crossing TRE-MYC mice with LAP-tTA mice where the liver activator protein (LAP) promoter drives expression of the tetracycline trans-activating protein (tTA) in liver cells66. As shown in Fig. 4, we have observed in LAP-tTA/TRE-Myc mice that liver tumors develop at 3 wks after removing doxycycline water to activate Myc gene (Figure 7A). When mice with liver tumors were exposed to doxycycline again, the size of liver tumor was noticeably reduced after 4 wks (Figure 7B), suggesting c-Myc is critical in liver tumor initiation and development in these mice. Histological analyses have shown that the Myc-induced tumors resembled HCC or hepatoblastomas66. **Treatment and tumor monitoring**: TRE-MYC/FVB mice are crossed with LAP-tTA/mice (Jackson Laboratory, Bar Harbor, ME) which have been backcrossed with FVB mice for more than 10 generations in our lab. When male mice are 8 wks old, doxycycline will be removed from drinking water. Three weeks after the doxycycline removal, these mice will be subject to MRI imaging to determine the status of tumor growth. The size of tumor measured by T2-weighted MRI imaging, which will be acquired on a 7 Tesla MRI BioSpec system (Bruker, Billerica, MA), will be used to randomize the mice to sham or of biofield therapy treatment as illustrated in [Figure 9][1]. Again, these mice will continually be treated for 3 weeks (3 times per week, 30 min). When the mice show signs of the morbidity (hunched posture or labor breathing), these mice will then be euthanized, blood and liver (normal and tumor tissues) will be collected and fixed. If the treatment is working, it is unlikely that the animals will reach this stage of forced euthanizing. However, even if the absence of morbidity, we may choose to euthanize a subset of the treated animals to pathological examine them. The presence of abdominal metastasis such as extrahepatic tumor mass infiltrating neighboring organs, significant lymph node enlargement, or peritoneal carcinomatosis or lung mets will be examined. The Kaplan-Meier survival curve will be constructed to determine the median survival time and to document the impact of biofield therapy treatment in mutant cMyc-driven mouse HCC. EEGs will be collected from the therapists at four time points across the 5-week experiment. **Behavioral Tracking**: For the first animal study with mutant cMyc-driven mouse HCC behavioral tracking is done with GoPro cameras mounting outside the back of the cages. Subsequent studies animal tracking and movement will be conducted with the EthoVision XT software, Color GigE Cameras, a Desktop Workstation, and custom Observation Chambers. The EthoVision video tracking software will be used to track animal behavior from both the top view. We will record animals and the software is able to record videos as well as conduct live tracking, which is important for data preservation and review. [1]: https://mfr.osf.io/export?url=https://osf.io/wcx4h/?direct&mode=render&action=download&public_file=False&initialWidth=695&childId=mfrIframe&parentTitle=OSF%20%7C%20Figure%209.jpg&parentUrl=https://osf.io/wcx4h/&format=2400x2400.jpeg