Dear Professor Kausikdatta:

We greatly appreciate your helpful comments regarding our manuscript, and our point-by-point responses to each comment are outlined below.

1. Given that classical acupuncture was developed - and acupuncture points mapped out - for humans, how appropriate is the use of the two acupoints on rats in this study? How were their precise locations determined for use, and where is the evidence that those acupoints on the rats are functionally equivalent to the described acupoints in humans?

Response: The location of acupoints on rats/mice is the foundation of experimental research for acupuncture. We located the acupoints on the rats according to a classic rat map, which was drawn at Nanjing Agricultural University in 1991 according to the experience of Chinese veterinarians and experimental studies. We cannot say for certain that the acupoints on the rats are perfectly functionally equivalent to the described acupoints in humans. Although we observed the protective effects of acupuncture after brain irradiation on rats, clinical trials are required to determine if this effect could be reproduced in humans.

2. Given that electro-acupuncture (EA) as a modality was unknown in traditional Chinese medicine, how appropriate is its use as a substitute for standard acupuncture? Are the actions and effects of standard acupuncture and EA done at the same acupoints different from each other? Is there any evidence to that effect? Was this assessed for the purpose of this study?……I have a few more questions about the specificity of EA.

Response: Electric acupuncture has been widely used in Chinese clinical work. Some researchers have focused on the differences between manual and electric acupuncture (J Altern Complement Med. 2015;21(3):113-28; J Tradit Chin Med. 2014;34(6):754-8), but this was not the purpose of the present study. Different manual manipulations and electrical parameters exert different therapeutic acupuncture effects, which are closely related to the characteristics of the diseases being treated.
We did not observe any effects on the normal rats after electric acupuncture in this study, but some research using electric acupuncture-pretreatment has shown that electric acupuncture may activate STAT3 and MCPIP1 (Brain Res. 2013 Sep 5;1529:154-64; J Neuroinflammation. 2013 May 10;10:63). Unfortunately, we did not detect these molecules in this study.

3. Was there a control where the same amount of electricity was applied to the acupoints without using the acupuncture needles?

Response: We appreciate the excellent suggestion; however, we did not design such a control group in this study. In the future, we will consider adding this type of control. Indeed, some research has shown similar effects between electric acupuncture and transcutaneous electrical nerve stimulation (Int J Clin Exp Med. 2015 Jan 15;8(1):1156-61).

4. Was there a control where the acupuncture needles were placed in acupoints different from the two used and electricity was applied?

Response: In the present study, we only placed the acupuncture needles at the Baihui and Shuigou points. In another study (unpublished), we used the Baihui and Zusanli points, and observed protective effects. However, we did not compare the differences between these two combinations. Thanking you for your suggestion and that we will consider comparing them in the future.

5. In human patients, when the acupuncture needles are inserted, they are often rotated until the patient expresses the sensation of warmth or mild pain/discomfort - which the acupuncturists define as 'de qi', taken to be indicative of the correct placement of the needle required for successful treatment. How is this assessed in a rat? The results that the authors have presented appear to show undeniably that following EA, the levels of most variables tested have come down to the levels of naive controls, compared to the irradiated group. I would like to understand if this observed effect is specifically due to acupuncture, due to EA as a modality, or due to non-specific phenomenology associated with transdermal or transcutaneous electrical stimulation.

Response: In the present study, we lifted, thrust, and rotated the needles before electric stimulation. However, it is hard to determine whether the rats experienced “de qi.” This is the main limitation in animal studies of acupuncture. We believe that both manual and electric acupuncture exert their effects by stimulating the nerve (Nat Neurosci. 2010;13(7): 883-8; Nat Med. 2014;20(3):291-5). Therefore, there are no essential differences between the two methods; only the manner in which the nerve is stimulated is different. Currently, it remains unclear which stimulation method is most suitable for treating brain irradiation injuries.

Sincerely,

Prof. Kai-Liang Wu
Department of Radiation Oncology
Fudan University Shanghai Cancer Center
Shanghai, China
Tel.: (8621) 64175590-6725
Fax: (8621) 6443 9052
E-mail: wukailiang@aliyun.com