Author's response to reviews Title:Acupuncture at homotopic acupoints exerts dual effects on bladder motility in anesthetized rats Authors:

1. Background: “It is well-known that dual effects of acupuncture on the 32 bladdercan be generated based on different contractile states of the bladder, However,the dual effects of single acupoint stimulation and acupoint site-specificity on thebladder have yet to be investigated.”Answer: We revised the sentence as: It is well-known that dual effects of acupunctureon the bladder can be generated based on different states of the bladder, however, thedual effects of single acupoint stimulation and acupoint site-specificity (homotopicacupoints and heterotopic acupoints) on the bladder have yet to be investigated.” 2. Conclusions: “MA at homotopic acupoints may produce dual effects on bladdermotility that is dependent on the bladder’s contractile state.”Answer: We revised the sentence as “MA at homotopic acupoints may produce dualeffects on bladder motility: inhibiting bladder motility when in an active state andenhancing bladder motility when in a static state.” Background.1. At the end of the second paragraph: “Conversely, acupuncture at heterotopicacupoints, which utilize different spinal segments to innervate a visceral organthan the where the afferent signal originates, has been shown to inducegastrointestinal –peristalsis in complete spinal rats.”Answer: We revised the sentence as “Conversely, acupuncture at heterotopic acupoints,which utilize different spinal segments to innervate a visceral organ has been shown toinduce gastrointestinal facilitation in complete spinal rats”. Methods – Intravesical pressure recording. – Need to be specifically addressed!1. It is still NOT clear to me how the different contractile states (active vs static) ofthe bladder is achieved and maintained in this experiment. Did the authors dosomething to create and maintain these states, OR, does the bladder go intothese states by themselves? Since the state of the bladder is critical to thehypothesis and experiments in this paper, it is critical for the authors to explainhow different states of the bladder are maintained. I could not find the answer tothis question anywhere in this draft.Answer: In “methods”, we mentioned “A manometric balloon was inserted into thebladder, filled with approximately 0.5 mL warm water, and was connected topolyethylene tubing, providing a pressure of approximately 100 mmH2O”. At this pressure, the bladder’s active and static state automatically switched in between them.We did not further control the states through increasing water or decreasing water.Results – much improved, suggestion below is not mandatory1. The last paragraph is titled “MA produced dual effects on the intravesicalpressure,” while the majority of this paragraph talked about bladder motility (MI). Isuggest adding “and motility” to the title sentence and also add a Part B) toFigure 3, in a similar format as the original Figure 3, but plot MI on the verticalaxis rather than bladder pressure. This way, your paragraph will match its title aswell as its picture.Answer: We thank you for your suggestion. As we mentioned in the response to yourcomment we submitted last time, intravesical pressure is the parameter whichrepresents the bladder motility. In another word, the bladder motility could be expressedthrough the intravesical pressure. This is why we use intravesical pressure in the titlehere. If we add “and motility”, that will be repetitious. Figure 3 is the pooled data shownin Figure 1 and 2. We pooled them together to show clearly which acupoint has dualeffects on bladder motility, not new data. Discussion – overall much improved, except for the very last sentence.1. Last sentence: “Here, we would also like to note that the results of acupunctureat the heterotopic acupoints in hind limbs were different than a previous report[42]. These differences might be due to the degree of 301 bladder distension(half-filling of the bladder in this study vs how much in the other study) or differentstimulations (MA vs electrical??).” Answer: The differences between our experiment and the reference 42 are as thefollows: (1) In our experiment, we used manual acupuncture; but they usedelectrostimulation. (2) In our experiment, the bladder pressure is approximately 100mmH2O at which the bladder’s active and static state automatically switched in betweenthem. But in their experiment, the pressure was changed through increasing ordecreasing water from 100 mmH2O to maximum of 300 mmH2O. (3) In our experiment,MA at heterotopic acupoint didn’t produce excitatory or inhibitory effect when thebladder is active but produced excitatory effect when the bladder is static. In theirexperiments, stimulating hindlimb (similar to heteropic acupoints) could induceexcitatory and inhibitory effects when the bladder pressure was low or high. To knowthe differences well, we also provided additional references as the below.References:1. Morrison JFB, Sato A, Sato Y, Suzuki A. The nitric oxide synthase inhibitor L-NAME reduces inhibitory components of somato-vesical parasympatheticreflexes in the rat. Neurosci Res 1996, 24: 195–199.2. Morrison JFB, Sato A, Sato Y, Yamanishi T. The influence of afferent inputs fromskin and viscera on the activity of the bladder and the skeletal musclesurrounding the urethra in the rat. Neurosci Res 1995, 23: 195–205.