Introduction: Human-in-the-loop optimization has made great progress to improve the performance of wearable robotic devices and become an effective customized assistance strategy. However, a lengthy period (several hours) of continuous walking for iterative optimization for each individual makes it less practical, especially for disabled people, who may not endure this process. Methods: In this paper, we provide a muscle-activity-based human-in-the-loop optimization strategy that can reduce the time spent on collecting biosignals during each iteration from around 120 s to 25 s. Both Bayesian and Covariance Matrix Adaptive Evolution Strategy (CMA-ES) optimization algorithms were adopted on a portable hip exoskeleton to generate optimal assist torque patterns, optimizing rectus femoris muscle activity. Four volunteers were recruited for exoskeleton-assisted walking trials. Results and Discussion: As a result, using human-in-the-loop optimization led to muscle activity reduction of 33.56% and 41.81% at most when compared to walking without and with the hip exoskeleton, respectively. Furthermore, the results of human-in-the-loop optimization indicate that three out of four participants achieved superior outcomes compared to the predefined assistance patterns. Interestingly, during the optimization stage, the order of the two typical optimizers, i.e., Bayesian and CMA-ES, did not affect the optimization results. The results of the experiment have confirmed that the assistance pattern generated by muscle-activity-based human-in-the-loop strategy is superior to predefined assistance patterns, and this strategy can be achieved more rapidly than the one based on metabolic cost.