OBJECTIVE: To evaluate the performance of a newly developed intelligent acupuncture robot that simulates human manual acupuncture techniques through an animal experiment using miniature pigs. METHODS: Two 3-month-old Bama miniature pigs were selected. One pig was used for the manual needling techniques of the practitioner, and the other pig was used for the intelligent acupuncture robot. The acupoints selected were "Qiangfeng" "Bojian" "Xiaokua" "Huiyang" (BL 35) and "Baihui" (GV 20), with a straight insertion depth of 25-35 mm. The manipulation techniques, including lifting-thrusting and twisting at a frequency of 90 times/min, were applied for 1 min, with 30 s each for lifting-thrusting (10 mm amplitude) and twisting (180° angle). The practitioner's needling techniques were captured using an optical motion capture system, and the collected data were input into the intelligent acupuncture robot for replication on the second miniature pig. A complete needling session of five acupoints was considered one round, and three rounds were performed per session, every other day, for a total of five sessions, completing 15 rounds. The Fréchet distance method was used to compare the robot's replicated needling data with the practitioner's data, and curve fitting analysis was conducted to evaluate the robot's performance and precision. RESULTS: The intelligent acupuncture robot achieved a total completion rate of 96.00% for 75 needling operations, with a non-completion rate of 4.00%. The robot's replication of the practitioner's needling techniques showed a good fit, with many characteristic points overlapping. The average Fréchet distance was 18.67. The Fréchet distance for the lifting-thrusting and twisting techniques at Baihui (GV 20) and "Huiyang" (BL 35) acupoints was smaller than that at "Xiaokua" and "Qiangfeng" points (P<0.01). The accuracy of the lifting-thrusting technique at "Xiaokua" and "Qiangfeng" acupoints was lower than the twisting technique (P<0.01). CONCLUSION The intelligent acupuncture robot is able to replicate the pre-recorded manual needling parameters with a high degree of accuracy.
Animals ; Swine ; Swine, Miniature ; Acupuncture Therapy/methods* ; Robotics/instrumentation* ; Humans ; Acupuncture Points ; Female ; Male

Circular RNAs (circRNAs) are a new class of non-coding RNAs, which have been confirmed to regulate insect gene expression and immune response through multiple manners such as competing endogenous RNA (ceRNA) regulatory network. Currently, function of circRNA in honey bee immune response remains unclear. In this study, PCR and Sanger sequencing were performed to validate the back splicing (BS) site of ame_circ_000115 (in short ac115). RT-qPCR was used to detect the expression profile of ac115 in larval guts of Apis mellifera ligustica stressed by Ascosphaera apis. Dual-luciferase reporter gene assay was conducted to verify the binding relationship between ac115 and ame-miR-13b. Interference of ac115 in larval guts was carried out by feeding specific siRNA, followed by determination of the effect of ac115 interference on expression of six genes relevant to host immune response. The results confirmed the existence of BS site within ac115. Compared with the un-inoculated group, the expression of ac115 in 4-day-old larval gut of the A. apis-inoculated group was up-regulated with extreme significance (P < 0.000 1), while that in 5- and 6-day-old larval guts were significantly up-regulated (P < 0.05). The brightness of specific band for ac115 in 4-, 5- and 6-day-old larval guts of the siRNA-circ_000115-fed group gradually became weak, whereas that of the siRNA-scrambl-fed group was pretty high without obvious variation. Compared with that of the siRNA-scramble-fed group, the expression of ac115 in 4-day-old larval gut of the siRNA-circ_000115-fed group was significantly down-regulated (P < 0.05), whereas that of the 5- and 6-day-old larval guts were down-regulated with extreme significance (P < 0.001). Ame-miR-13b was truly existed and expressed in A. m. ligustica larval guts, and there was true binding relationship between ac115 and ame-miR-13b. Compared with that of the siRNA-scramble-fed group, the expression of antimicrobial peptide genes hymenoptaecin and abaecin in 6-day-old larval gut of the siRNA-circ_000115-fed group was significantly up-regulated (P < 0.05), while that of ecdysone receptor (Ecr) was down-regulated with extreme significance (P < 0.01). These results indicate that ac115 is truly expressed in A. m. ligustica larval guts, BS site truly exists within ac115, and effective interference of ac115 in A. m. ligustica larval guts can be achieved via feeding siRNA. Moreover, ac115 potentially regulates Ecr expression through adsorption of ame-miR-13b and expression of hymenoptaecin and abaecin using a non-ceRNA manner, further participating in host stress-response.
Bees/genetics* ; Animals ; Larva/metabolism* ; RNA, Circular/genetics* ; RNA, Small Interfering/genetics* ; MicroRNAs/genetics*