Background: Vertex-vertex and vertex-nonvertex presentations are the most common and advisable for vaginal delivery. We aimed to explore the maternal and neonatal outcomes of twin pregnancy between vertex and non-vertex presentations of the 2nd twin in vaginal delivery. Methods: In this unicentric retrospective cohort study, data of 213 mothers during twin pregnancy after vaginal delivery from January 2016 to July 2020 were collected. The women were divided into the vertex-vertex presentation group (VV) and vertex-breech presentation group (VB). Data of maternal and neonatal outcomes were compared between groups. Results: Among the 213 mother-infant dyads, there were 140 women in the VV group and 73 women in the VB group (65.73% vs. 34.27%). There were more males for the 1st fetus (60.3% vs. 35.7%, P=0.001), less twin-to-twin delivery interval (12.260±10.437 min vs. 9.360±6.509 min, P=0.032), less birthweight for the 2nd fetus (2218.160±42.698 g vs. 1999.040±66.288 g, P=0.045), greater birthweight discordance (233.814±16.576 g vs. 273.260±31.688 g, P=0.024), earlier gestational age (243.214±1.626 day vs. 235.2055±2.455 day, P=0.030), more breech extraction used (98.6% vs. 0, P<0.001) in the VB group. Infants in the VB group had a higher incidence of admission to NICU (P=0.017), lower 1-minute (P<0.001) and 5-minute Apgar score (P=0.043) for the 2nd twin. After the adjustment of the twin gender, birth weight, chorionic properties and gestational age, the 2nd twin in the VB group still had lower 1-minute Apgar scores. Adjusted linear regression showed that twin presentation remained associated with gestational age (P=0.045).Key Conclusions: The 2nd twins of the vertex-breech presentation have lower 1-minute Apgar scores compared with those in the vertex-vertex presentation. Fetal presentation is an independent factor for 1-minute Apgar score. Fetal presentation is associated with increased odds of preterm. However, further exploration of this relationship is still needed.