<p>This study aimed to assess the skin temperature (ST) change at the acupoint and the corresponding meridian when needling based on the “moving Qi and circulating Blood” effect of the bilateral BL65 acupoints, the Shu-stream acupoints recorded in ancient literature. A non-randomized, self-controlled pilot study conducted a 30-min acupuncture intervention at bilateral BL65 acupoints on thirty healthy volunteers. The ST was measured on two regions: the cutaneous region of the Bladder meridian (BMCR), and the cutaneous region not belonging to the Bladder meridian (non-BMCR) by an infrared thermal image camera at five junctures. The results showed that the ST at the BMCR significantly increased during needling at the BL65 acupoint (<i>p</i> = 0.03). In contrast, there was no significant change in the ST at the non-BMCR (<i>p</i> = 0.76). In conclusion, acupuncture could specifically increase the ST at the cutaneous regions of the Bladder meridian when needling at BL65 acupoints (Shu-stream acupoints). This effect could refer to the meridian theory and the literature-recorded “moving Qi and circulating Blood” effect of the Shu-stream acupoints.</p>